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Volume 32,2024 Issue 10

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Effects of different transplantation routes and fecal status on the structural abundance of gut microbiota

LIU Kaihui, GUO Yaxi, DU Xiaopeng, WANG Zhaohua, ZHU Hua

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【Objective】To evaluate the effects of the fecal status and transplantation method on intestinal flora structure after fecal microbiota transplantation in germ-free mice. 【Methods】Thirty-six C57BL/6 mice were randomly divided into three groups: fresh fecal gavage transplantation group, frozen fecal gavage transplantation group and frozen fecal rectal transplantation group. Feces were collected at 2 weeks, 4 weeks and 6 weeks after transplantation, and all mice were sacrificed at 6 weeks to obtain the contents of the small intestine and the large intestine. The structure and function of the gut microbiota and the dynamic trends of microbial changes were analyzed by 16S rRNA sequencing. 【Results】By α-diversity analysis, compared with the frozen fecal gavage transplantation group, the diversity of the small intestine microbes in both the fresh fecal gavage transplantation group and the frozen fecal rectal transplantation group showed no significant differences (P > 0.05), but the diversity of large intestine microbes was significantly increased (P < 0.001). By β-diversity analysis, samples from the small intestine of the fresh fecal gavage transplantation group and the frozen feces gavage transplantation group clustered in the same area, indicating that the microbial community composition was similar (P > 0.05). However, samples from the large intestine were distributed in different areas, showing significant differences (P = 0.001). For the frozen fecal gavage transplantation group and the frozen fecal rectal transplantation group, samples from both the small intestine and the large intestine were distributed in different areas, with significant differences (P = 0.001). By LEfSe analysis, Bacteroidota was relatively dominant in the fresh fecal gavage transplantation group, while Verrucomicrobiota and Proteobacteria were relatively dominant in the frozen fecal gavage transplantation group, and Firmicutes were relatively dominant in the frozen fecal rectal transplantation group. Functional prediction by PICRUSt2 showed that neither the fecal status nor the method of transplantation induced changes in the microbial community's functions. 【Conclusions】Compared to frozen fecal gavage transplantation, both fresh fecal gavage transplantation and frozen fecal rectal transplantation enhance the diversity of microbes in the large intestine. The fecal status does not affect the gut function and colonization trends of the microbiota, whereas the method of transplantation affects the colonization trends but not the functions of the microbiota.
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Comparison study of two plasmid vectors expressing human thyroid stimulating hormone receptor inducing mice model of Graves' disease via electroporation

Lin Xiaoying, Zhang Meng, Zhou Xingchen, Wu Mengzhi, Xv Huayang, Wang Ling, Wu Liping, Shi Bingyin

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Objective The purpose of this study was to provide a more effective method for the research of prevention and treatment of Graves’ disease by comparing the effects of two plasmid vectors expressing the human thyroid stimulating hormone receptor (TSHR) A subunit gene in inducing animal model of Graves" disease via electroporation. Methods pcDNA3.1-THSR A and pTriEx1.1-THSR A expressing TSHR A subunit were constructed and used to induce Graves" disease by intramuscularly injecting with immediate electroporation once every 3 weeks for a total of 4 times. Mice in thecontrol group were injected with PBS. One week post-second electroporation, blood was gained to measure serum thyrotropin receptor antibody (TRAb). Three weeks post-last electroporation, echocardiography was performed on mice. Mice were sacrificed 4 weeks after the last electroporation, blood, thyroid, orbital tissues were collected, serum total thyroxine (TT4) was measured, and histological examination was performed. Results The average value of serum TRAb in the pcDNA3.1-TSHR A group (n=15) and the pTriEx1.1-TSHR A group (n=13) was (6.9±2.0) U/L and (7.5±2.2) U/L, respectively. The latter was significantly higher than that in the control group (4.9±0.5) U/L (P＜0.05). The average value of serum TT4 in the pcDNA3.1-TSHR A group and the pTriEx1.1-TSHR A group was (41.4±23.8) ng/ml and (63.2±53.7) ng/ml, respectively, both higher than the control group (20.2±4.0) ng/ml (P<0.01). Still, there was no significant difference between thetwo experimental groups (P=0.152). Thyroid pathology showed thyroid follicular epithelial hyperplasia with T-cell infiltration in themodel group. Echocardiography showed that levels of left ventricle mass in the pTriEx1.1-TSHR A group were higher than that in the control group (P＜0.01) and the pcDNA3.1-TSHR A group (P＜0.05). Orbital pathology showed fibrotic changes in the extraocular muscles of mice in the model groups. Conclusions Both pcDNA3.1 and pTriEx1.1 expressing TSHR A subunit were able to induce Graves" disease in mice by electroporation, and the efficiency of two plasmids expressing TSHR A subunit in inducing hyperthyroidism and Graves’ ophthalmopathy was similar. The efficiency of pTriEx1.1-TSHR A inducing thyrotoxic heart disease is better than pcDNA3.1-TSHR A.
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Research progress of motor function evaluation methods in animal models of sarcopenia

Yang Sen, Zhang Yong, Zhou Zhixiong, Zeng Ping

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Sarcopenia is an age-related skeletal muscle degenerative disease. The physiological aging mice are the most commonly used animal model for studying sarcopenia. Since sarcopenia is characterized by decreased skeletal muscle mass and reduced muscle strength, the exercise performance is a reflection of muscle function and widely used to evaluate sarcopenia. The motor function evaluation for sarcopenia mice model is generally designed based on muscle endurance, muscle strength, coordination and balance. The methods include run table exhaustion, voluntary wheel, grip strength, the horse grid, bars and balance beam test, etc. By collating recent publications, we systematically summarized the methods for motor function evaluation, including the test principle, test procedure, evaluation indexes, advantages and disadvantages, and then proposed the operational program for evaluation of sarcopenia phenotype, which will be of help for researchers to choose appropriate evaluation methods according to their specific research purposes. Furthermore, the innovative technology of motor function assessment was summarized, which would be instructive in evaluation of skeletal muscle function and diagnosis of sarcopenia.
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Application status of model biological zebrafish in radiation medicine research

DONG Juancong, WANG Jingjie, YU Jinhuan, CHENG Jiao, DANG Xuhong, YANG Guodong

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As a model organism between cells and traditional mammals, zebrafish has shown broad application prospects in radiation medicine research with its unique advantages. It has the characteristics of high homology with human genes, high fertility, short embryonic period, transparent and easy to observe embryos, making it an important tool in radiation medicine research. Recently, zebrafish has made remarkable progress in the application of low dose radiation biological effects, radiation therapy and radiation damage prevention and treatment, and they are the key areas of radiation medicine. In this paper, these applications are reviewed in order to explore the potential value of zebrafish in radiation medicine research and provide reference for experimental research in related fields.
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Characterization of intestinal flora and transcriptomics in rats with gastric cancer

ZHANG Chenxi, LI Jijuan, ZHANG Feicheng, GAO Tianyu, LIANG Xinyue, PAN Lijia

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Objective To analyze the differences in intestinal microbiota and transcriptomics between N-methyl-N'-nitro-n-nitrosoguanidine (MNNG) gastric cancer rats and normal rats, in order to explore the intestinal microbiota and transcriptomic characteristics of MNNG gastric cancer rats, and to analyze the correlation between the two, so as to provide a reference for related studies using MNNG gastric cancer rats as a model. Methods A total of 12 Wistar rats were randomly divided into normal (NM) and gastric cancer (GC) groups, the gastric cancer group was given a concentration of 20 mg/mL of MNNG by gavage, the dose was 100 g/mL, once a day, and the normal group was given the same dose of normal saline gavage, and the samples were collected for testing after 16 weeks of continuous intervention. The gastric tissues of rats were collected and stained by HE staining to observe the morphological changes of the gastric mucosa of the two groups, and the expression levels of differential genes were detected by transcriptome sequencing. Sterile EP tubes collect cecal contents for 16S rRNA sequencing. Results 1.Macroscopic observation and HE results: The volume of gastric tissue mucosa in the NM group was normal, the surface was shiny, the gastric wall was elastic, the mucosal folds were regular, there was no hyperplasia and no bleeding points, and the gastric mucosa volume of the gastric tissue in the GC group was reduced, the gastric wall was thinned, the elasticity was poor, the folds were disordered and irregular, and there was a bulge accompanied by yellow-black keratinosis. HE staining, the squamous epithelial layer, submucosal layer and muscular layer of the gastric mucosa in the NM group were clear and there was no proliferation and keratosis, while the gastric mucosal layer and cell polarity disorder in the GC group were different, the cell morphology was different, the squamous epithelial layer was destroyed, the squamous epithelial cells proliferated and keratinized, and proliferated downward to invade the muscle, and the modeling was successful. 2. The results of intestinal microbiota sequencing showed that the abundance of Akkermansia and Lactobacillus in MNG gastric cancer rats decreased significantly, and the abundance of Rumen coccaceae, Prevonella and Blauter increased significantly. 3. The three key pathways obtained by transcriptomic sequencing KEGG pathway enrichment analysis were amebiasis, systemic lupus erythematosus and PI3K-AKT signaling pathway, and the five differential genes enriched to these three pathways were MCPT8I2, IGH-6, IGHG1, ACTN2 and VEGFD. 4. The results of combined analysis of intestinal microbiota and transcriptomics showed that _UCG-005, Prevonella _UCG-003 and Brautella may be positively correlated with amebiasis, systemic lupus erythematosus and PI3K-AKT signaling pathway. Conclusions The abundance of intestinal microbiota in gastric cancer rats formed by MNNG gavage is different from that of normal rats, and the up-regulated MCPT8I2, IGH-6, IGHG1, ACTN2 and VEGFD may be the differential genes of gastric cancer induced by MNNG gavage, and the combined analysis of intestinal microbiota and differential genes suggests that the mechanism of MNNG carcinogenesis may be mainly related to the destruction of gastric mucosa and the inflammatory response.
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Establishment and evaluation of rat model of type 2 diabetes with liver fibrosis

HE Hongxing, LIAO Naishun

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Objective To establish a rat model of type 2 diabetes mellitus with hepatic fibrosis by using the high-lipid-sugar diet (HLSD) combined with the inducement of CCl4 and streptozotocin. Methods SD rats were randomly divided into liver fibrosis (LF) group, type 2 diabetes combined with liver fibrosis (DLF) group, and normal group, 10 rats in each group. All rats in LF or DLE group were fed with HLSD for 4 weeks, and then all rats were intraperitoneally injected 20% carbon tetrachloride (CCl4, once in a week) according to the body weight of 5 mL/kg to induce hepatic fibrosis for another 4 weeks. During the CCl4 treatment, rats in the DLF group were also intraperitoneally injected with 25 mg/kg streptozotocin (STZ, once in a week). After STZ treatment, the content of FBG, INS, HOMA-IR, and HbA1c were evaluated once in a week. The FBG level was detected by blood glucose meter. The contents of INS and HbA1c were detected by ELISA method. Biochemical analysis was used to analyze the serum levels of AST, ALT, Tbil, and HA, as well as the HYP content in liver tissues. HE staining and Masson staining were used to evaluate the pathological changes in liver tissues. Results The contents of FBG, HbA1c and HOMA-IR in DLF group were significantly increased as compared to those in normal or LF group. While the INS content in DLF group was significantly decreased when compared to those in normal or LF group. Comparing with normal group, the high levels of ALT, AST, Tbil, HA and HYP, as well as fibrotic changes and inflammation in liver tissue were all observed in LF group and DLF group. Conclusions HLSD combined with CCl4 and streptozotocin inducement is an effective strategy to establish a rat model of type 2 diabetes mellitus with liver fibrosis.
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The mechanism of airway remolding in mice model of chronic obstructive pulmonary disease induced by cigarette smoke combined with Klebsiella pneumoniae

ZhangZeyu, MEIXiaofeng, TAOLiuying, LIULan, LIJiansheng, ZHAOPeng

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Purpose: Cigarette smoke exposure combined with Klebsiella pneumoniae (KP) infection were used to establish the model of chronic obstructive pulmonary disease (COPD) in mice to investigate the mechanism of airway remodeling. Methods: Male BALB/c mice (n=96) were randomly divided into Control group, CS group, KP group and CS+KP group. The mice were exposed to CS, KP and CS+KP from week 1 to 8, and were sacrificed at week 4, 8, 16 and 24. Minute volume (MV), the validity of the enhanced Pause (Penh), mean linear intercept (MLI), mean alveolar number (MAN) and the changes of lung pathological structure were detected. The expression levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in lung tissue were detected by ELISA kit. Collagen deposition was observed by masson staining and immunohistochemistry. The α-smooth muscle actin (α-SMA) and transforming growth factor-β1 (TGF-β1) expression in lung tissue were detected by immunofluorescence. Human bronchial epithelioid cells were stimulated by cigarette smoke (CS) and lipopolysaccharide (LPS). The expression levels of airway epithelial junction proteins, autophagy-related protein and mTOR signaling were detected. Results: Compared with the Control group, MV of mice in CS+KP group were significantly decreased from week 4 to 24 (P<0.05 or P<0.01), and Penh of mice in CS+KP group were significantly increased from 8 to 24 weeks (P<0.05 or P<0.01), MV were significantly decreased and Penh were significantly increased in CS group from 8 to 16 weeks (P<0.05 or P<0.01), MV were significantly decreased in CS group from 8 to 16 weeks. Compared with the Control group, MAN were significantly decreased and MLI were significantly increased in lung tissue of mice in CS+KP group from week 4 to 24 (P<0.05 or P<0.01). At the week 8, massive inflammatory cell infiltration, alveolar wall thickening, alveolar rupture and fusion, and airway wall thickening were observed in lung tissue of mice in CS+KP group from week 8 to 24. The expression levels of IL-1β and TNF-α in CS+KP group were significantly increased (P<0.05 or P<0.01) from week 4 to 24; massive inflammatory cell infiltration, alveolar wall thickened, alveolar rupture and fusion, and airway wall thickened were observed in the lung tissues of CS group and KP group from week 8 to 16. MAN was significantly decreased, MLI, IL-1β and TNF-α levels were significantly increased in the lung tissues of CS group and KP group from week 8 to 16 (P<0.05 or P<0.01). Col Ⅰ, col Ⅲ, α-SMA and TGF-β1 in lung tissue of mice in CS+KP group were significantly increased from week 8 to 16(P<0.01); col Ⅰ was significantly increased in CS group from week 8 to 16 (P<0.01); col Ⅰ were significantly increased in KP group from week 8 to 16 and col Ⅲ were significantly increased in KP group at 8th week (P<0.01). In addition, increase in E-cad and decrease in N-cad were observed in 16HBE cells induced by CS and LPS (P<0.05), LC3B and Beclin-1 expression were significantly decreased (P<0.05), the expressions of p-mTOR, p-P70-S6K, and p-4E-BP1 were significantly increased in 16HBE cells induced by CS and LPS (P<0.05 or P<0.01). Conclusion: Pulmonary function declined, pathological changes of lung tissue and airway remodeling in COPD mice induced by CS and KP appeared to occur early and last long, and its mechanism may be related to the activation of mTORC1 signaling pathway to inhibit autophagy.
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Mechanisms of Shenlingcao oral liquid against non-small cell lung cancer by network pharmacology combined with molecular docking and experimental verification

LUO Jia xin, ZHAN Yang, SUN Den long, WU Zhen peng, LIU Yu qing, LIU Wen jun

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Objective In this study, we aimed to predict the inhibitory mechanism of shenlingcao oral liquid (SLC) on non-small cell lung cancer (NSCLC) by network pharmacology, and to verify it by molecular docking and in vivo animal experiments. Methods Active ingredients and corresponding targets of SLC and NSCLC were obtained by database and literature search. The common target of SLC and NSCLC was selected to construct the protein interaction network, and GO and KEGG enrichment analysis and molecular docking were performed. Lewis lung cancer mouse model was constructed, and Model, SLC high-dose group and SLC low-dose group, high-dose group and low-dose group were administered by intragastric administration at the dose of 8.75 g SLC lyophilized powder/kg and 3.50 g SLC lyophilized powder/kg, respectively. After 14 days of drug intervention, the tumor growth, pathological changes of tumor tissue and apoptosis of tumor tissue of tumor-bearing mice were observed, the changes of blood routine indexes of mice were detected, and the expression of p-AKT, AKT, p-PI3K, PI3K and Bcl-2 protein in tumor tissues of mice were detected, and the results of KEGG enrichment were verified. Results The network pharmacological analysis showed that there were 77 active ingredients, 618 potential targets, 1498 potential targets for NSCLC, 179 drug and disease intersection targets. Intersection target enrichment analysis showed that it was mainly concentrated in phosphatidylinositol 3 kinase signaling pathway (PI3K-AKTsignaling pathway), mitogen-activated protein kinase signaling pathway (MAPK signaling pathway) and other related pathways. Molecular docking showed that the top 10 core components had good bonding ability with the top 10 core targets. The results of animal experiments confirmed that compared with the Model, the tumor volume and weight in SLC high-dose and low-dose groups were significantly decreased (P < 0.05, 0.01), the expressions of white blood cells (WBC), neutrophils (Neu) and monocytes (Mon) were significantly decreased (P < 0.05, 0.01, 0.001), while the expressions of red blood cells (RBC), platelets (PLT) and hemoglobin (HGB) were significantly increased (P < 0.05, 0.01, 0.001), the apoptotic cells were significantly increased in early tumor tissue (P < 0.05, 0.01), and the protein expression levels of p-PI3K/PI3K, p-AKT/AKT and Bcl-2/GAPDH were significantly decreased (P < 0.05, 0.01). The expression levels of PI3K, AKT1 and Bcl-2 genes were significantly decreased (P < 0.05, 0.01, 0.001). Conclusion The mechanisms of SLC against NSCLC may be related to the activation of PI3K-AKT pathway and the promotion of apoptosis.
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Exploring the Mechanism of Action Huoxue Tongluo Prescription for the Treatment of Spinal Cord Injury Based on Network Pharmacology and Molecular Docking Technology

WANG Jiaxing, CHEN Jiayang, SHEN Tingting, WANG Zhihui, REN Xiaoping

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This study preliminarily investigated the potential mechanisms of the Huoxue Tongluo prescription(HXTLP) in treating spinal cord injury(SCI) through a combination of network pharmacology, molecular docking technology, and in vivo experiment verification. TCMSP database and Swiss Target Prediction platform were utilized to select the active ingredients and targets in HXTLP with therapeutic properties for SCI, and the "active ingredients - targets" network was construted.SCI related targets were searched in OMIM and GeneCards, and the protein interaction network (PPI) of the common targets of HXTLP and SCI was established based on the STRING database. The Metascape database was used for KEGG pathway enrichment and GO analysis of the common targets. Molecular docking of active ingredients and key targets was performed through Autodock1.5.7 software and the results were visualized by Pymol software. Finally, the effect of HXTLP on SCI was verified by animal experiments.The results indicated that A total of 184 intersection targets were obtained, and the key targets were AKT1, STAT3, HSP90AA1, PIK3CA, PIK3R1, HRAS, MAPK1, EGFR, etc. Molecular docking results showed a strong binding ability between the core active components and key targets. Animal experimental results showed that HXTLP group had significant therapeutic potential for SCI compared with model group. Behavioral scores and footprint analysis showed improved hind limb motor function, and histological examination showed more complete structure and morphology of the injured area.WB result revealed that HXTLP can effectively inhibit the key target protein HSP90AA1 and the expression of P-STAT3, promoted the expression of P-AKT1. This study verifies that HXTLP has the characteristics of multi-component, multi-target and multi-pathway synergistic effect in the treatment of SCI, and provides experimental theoretical basis for clinical medication and further research of SCI.
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Japanese encephalitis virus escape type I interferon mechanism in the creation of a mouse infection model

ZHOU Yifan, ZHANG Caiqin, LI Bingrun, BAO Jiaojiao, ZHANG Yanying, SHI Changhong

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Japanese encephalitis virus (JEV) usually evades the inhibitory effect of innate immunity type I interferon (I-IFN) when infects human cells and tissues, then causes a series of serious symptoms such as spasticity, neurodegenerative lesions, neuroinflammation and even death. Generally, JEV escapes innate immunity by inhibiting IFN-α/βproduction and interferon’ Janus kinase (Jak)-signal transducer and activator of transcription (STAT) signaling pathway. Because of this special immune escape mechanism, various mouse infection models have been constructed for the study of the pathogenesis and therapeutic regimen of JEV infection. In this review, based on the exposition the IFN immune escape mechanism of JEV, we systematically introduced the JEV infected mouse models, analyzed the characteristics of these models and the degree of simulation of human symptoms, etc. To sum up, we hope to develop various new JEV infected mouse models based on the new research targets may be found and provide novel ideal animal models for JEV research.
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Study on the molecular mechanism of the effect of phosphatidylcholine on dyslipidemia based on Apoa-I regulation of cholesterol reverse transport

ZHANG Qi, SUI GUOYUAN, SONG NAN, JIA LIANQUN

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[Abstract] Objective: Based on the regulation of cholesterol reverse transport by Apoa-I, to explore the effect of phosphatidylcholine on hyperlipidemic mice. Method: Thirty Apoa-I -/- mice were randomly divided into Apoa-I -/- group, Apoa-I -/-+HFD group, and Apoa-I -/-+HFD+PC group using a random number table method; 30 C57BL/6J mice were randomly divided into WT group, WT+HFD group, and WT+HFD+PC group as a control group, with 10 mice in each group. The Apoa-I -/- group and WT group were fed with basic feed, while the other groups were fed with high-fat feed for 8 weeks to establish a hyperlipidemia model. Starting from the 9th week, the WT+HFD+PC group and Apoa-I -/-+HFD+PC group were given PC 2.5 g·kg-1·d-1, while the remaining mice were given physiological saline by gavage for a total of 4 weeks of intervention. The serum lipid levels of mice were detected using a fully automated analyzer. Hematoxylin eosin (HE)staining and Oil red O staining was used to observe pathological and morphological changes in mouse liver tissue. COD-PAP method was used to detect cholesterol levels in mouse liver tissue. ELISA method was used to detect LCTA levels in mouse serum RT-qPCR and Western Blot methods were used to detect the mRNA and protein expression of cholesterol ATP binding cassette transporter A1,（ABCA1）, ATP binding cassette transporter G1（ABCA1）, lecithin cholesterol acvltransferase（LCAT）, Hepaticlipase（HL）, scavenger receptor class B type I（SR-BⅠ）, low density lipoprotein receptor（LDL-R） in liver tissue. RESULTS: Compared with the WT group, the serum lipid levels of mice in the WT+HFD group were significantly increased (P<0.01), hepatic fat vacuoles were obvious, hepatic lipid deposition was significant, and TC levels in liver tissue were significantly increased (P<0.01). The mRNA and protein expressions of ABCA1, ABCG1, LCAT, SR-B1, HL, and LDL-R were significantly increased (P<0.05, P<0.01). Compared with the WT+HFD group, the serum lipid levels of mice in the WT+HFD+PC group were significantly reduced (P<0.05, P<0.01), hepatic fat vacuoles were significantly reduced, hepatic lipid deposition was alleviated, and TC levels in liver tissue were significantly reduced (P<0.01). The mRNA and protein expressions of ABCA1, LCAT, SR-B1, HL, and LDL-R were significantly reduced (P<0.05, P<0.01); The serum levels of TC, TG, and LDL-C were significantly increased in the Apoa-I -/-+HFD group mice, while the levels of HDL-C were significantly reduced (P<0.05, P<0.01). Hepatocytes underwent balloon like transformation, liver lipid deposition was significantly aggravated, and TC levels in liver tissue were significantly increased (P<0.05). The mRNA and protein expressions of ABCA1, LCAT, HL, and LDL-R were significantly increased (P<0.05, P<0.01). Compared with the WT+HFD+PC group, the Apoa-I -/-+HFD+PC group mice showed a significant increase in serum lipid levels (P<0.05, P<0.01), significant hepatic lipid vacuoles, significant hepatic lipid deposition, and a significant increase in TC levels in liver tissue (P<0.05). The mRNA and protein expressions of ABCA1, ABCG1, LCAT, SR-B1, and HL were also significantly increased (P<0.05, P<0.01). Conclusion: Phosphatidylcholine can improve dyslipidemia in hyperlipidemic mice by interfering with Apoa-I and then regulating cholesterol reverse transport.
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Research progress in large - scale animal experimental research on medical devices

yang guang, gaoyang, cuiyixin, zhuhuaili, hujiawei, yangqian, cuichaoyue, weixufeng

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As China has become the second largest market for medical devices in the world, the domestic medical device industry has been growing. As an important part of preclinical evaluation of medical devices, large animal research directly affects the research and application of medical devices. Large animals are widely used in the evaluation of safety and feasibility of medical devices because they are closer to humans in terms of body size, anatomical structure and physiological functions. In large animal experimental research, the selection of suitable experimental animals and the establishment of suitable animal disease models are the basis for ensuring the smooth progress of experiments. In this paper, the selection of experimental animals and the establishment of disease models in medical device large animal experimental research are systematically sorted out, and the existing problems and deficiencies are pointed out.
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Advances in animal models of rheumatoid arthritis

WANG Junwei, GENG Xingchao, LI Wei

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Rheumatoid arthritis (RA) is a chronic autoimmune disease that significantly impacts joints. Experimental models are crucial tools for studying the pathogenesis, pharmacotoxicological mechanisms, and drug screening evaluation of RA. Currently, commonly used RA experimental models include animal models and in vitro models. With advancements in biotechnology and biomaterials, RA experimental models have evolved from induced and 2D in vitro models to spontaneous gene modification and 3D in vitro models. Some progress has been made in the study of traditional Chinese medicine (TCM) disease and syndrome combination models. This paper summarizes the research progress on modeling, monitoring, and evaluation methods of RA experimental models to provide reference for related research.
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Research progress on the preparation of animal models of non-alcoholic fatty liver disease and the combination of disease and syndrome

ma chun xia, chen zhen dong, tian xu dong

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Non-alcoholic fatty liver disease (NAFLD) is a metabolic syndrome caused by a variety of pathogenic factors and characterized by excessive deposition of fat in liver cells. At present, the pathogenesis of NAFLD is not fully understood, and there is a lack of specific drugs for NAFLD in clinical practice. Therefore, the establishment of an ideal animal model is extremely important for the clarification of the pathogenesis of NAFLD, the development of specific drugs and treatment. In the past five years, in addition to NAFLD animal models, with the continuous renewal and development of traditional Chinese medicine, there have been more and more disease-syndrome combined models related to NAFLD. This article summarizes the NAFLD animal models commonly used in the past five years, their preparation and evaluation methods, in order to provide reference for the future study of NAFLD animal model preparation and the combination of disease and syndrome.
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The role of the Camp signaling pathway in depression

liu jing xia

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Depression is a common mental illness at present,which brings a negative impact on the quality of life of patients, but also brings a serious economic burden to the society. In recent years, the strategy of interfering with depression based on signaling pathways has attracted widespread attention in society. As an important cellular signaling pathway, the cAMP pathway plays a key role in the pathogenesis and intervention of depression. Therefore, this review addresses recent studies on cAMP pathway-based interventions in depression, which provides a theoretical basis for regulating the cAMP pathway and its cascade mechanism on depression.
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The application progress of patient-derived organoids xenograft model in cancer precision therapy

XING Yanzi, ZHAO Jumei, SHI Changhong

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Precision therapy has become an important means of modern medicine, and its goal is to provide individualized treatment according to the characteristics of patients. The successful development of precision medicine depends on the application of preclinical cancer models. Patient-Derived Organoids (PDO) xenograft model display both characteristics of in vitro PDO model and in vivo Patient-Derived Tumor Xenograft (PDX) model. This model can not only maintain the heterogeneity of the original tumor, but also show more advantages, such as the large-scale cultivation, high throughput drug screening in vitro and drug sensitivity test in vitro. It is an innovative, precise preclinical disease model. In this review, we mainly summarize the basic characteristics of PDO xenograft model, analyze its construction methods and influencing factors, and further discuss its application in precision therapy. The aim is to provide a reliable preclinical experimental tool for cancer individual treatment.
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Research progress and application of SNP detection methods in genetic quality testing of laboratory animals

zhangyue, lixiangru, quanjinqiang, gaocaixia, xiachangyou, zhaoshengguo

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Laboratory animals are important experimental materials for life science research, and the uniformity of their quality plays a key role in the accuracy and reliability of the experimental results. Therefore, the genetic quality test of laboratory animals is an important link to evaluate the genetic quality of laboratory animals. With the progress of modern biotechnology, the methods of genetic testing for experimental animals have been updated. Single nucleotide polymorphisms (SNP) as molecular markers have been widely used in genetic testing of laboratory animals, and with the progress of modern biotechnology, the SNP detection methods for laboratory animals have been updated. In this review, we mainly sorted out the current research progress and application of SNP detection methods in genetic quality testing of laboratory animals, as well as the advantages and disadvantages of various detection methods, with a view to providing a reference basis for genetic quality testing of laboratory animals.
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Comparison of three Non-alcohol Steatohepatitis Models

XUE Jingbo, YANG Jinfeng, HUANG Kai, PENG Yuan, TAO Yanyan, LIU Chenghai

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Objective Compare the serological and pathological characteristics of three nonalcoholic steatohepatitis (NASH) models: high fat diet (HFD) with carbon tetrachloride (CCl4) injection, methionine and choline deficient diet (MCD) and Aymlin liver NASH (AMLN) diet induced NASH models, and make primary study on their pathological mechanisms. Methods Three NASH models were established by feeding HFD with CCl4 injection for 10 weeks, MCD for 8 weeks and Amylin diet for 26 weeks. After feeding, serum transaminase (ALT, AST) and glucose (GLU), liver triglyceride (TG), total cholesterol (TC), Superoxide dismutase (SOD) and Malondialdehyde (MDA) levels were measured. Insulin level was measured by ELISA and HOMA-IR index was calculated. Hematoxylin-eosin (HE) staining, Sirius red staining and Oil Red staining were used to indicate the pathological changes of liver. Furthermore, NAS score was used to evaluate the pathology grade. Results Compared to normal control mice, all mice in three model groups had obvious increase in serum transaminase, liver TG, TC, SOD, MDA level. The level of serum FINS, GLU and HOMA-IR index were significantly increased in mice of Amylin and HFD with CCl4 group, but decreased in mice of MCD group. According to HE staining, Oil Red staining and NAS score, all mice in three groups MCD mice had NASH phenotype changes. Liver collagen deposition was most obvious in mice of HFD with CCl4 group. Meanwhile, liver lipid droplets were most abundant in mice of AMLN group. Conclusions All three NASH models show different advantages and disadvantages. NASH models induced by Amylin diet recaptures the key NASH features observed in patients.
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Establishment of an animal model of varicose veins of lower limbs in rats

He Zhiyong

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Objective To construct a nearly clinical animal model of lower limb varicose veins in rats through surgery, providing theoretical support for evaluating drug therapy. Method 30 SD rats, half male and half female, were randomly divided into a control group and a surgical group. In the operation group, the rats' lower limb veins ( including small saphenous vein and femoral vein ) were ligated by the improved lower limb vein ligation operation, that is, the small saphenous vein was completely ligated with femoral vein, and thrombosis resulted in a lasting increase in the internal pressure of the deep veins of the lower limb, thus causing varicose symptoms of the lower limb. At the 6th week after surgery, the varicose veins of the rats in the surgical group were scored to select the rats that were successfully modeled.Then, the successfully modeled rats were randomly divided into a model group and a treatment group: Maizhiling 62.5 mg/kg was orally administered once a day, while the control group and model group received equal volume of physiological saline orally every day for 20 consecutive days. On the day before administration, 7 days, 14 days, and 20 days after administration, macro photography and scoring were performed on the lower limbs of rats. After completion, the model group and the Maizhiling group took approximately 1 cm long saphenous vein above the ankle joint of the lower limb on the surgical side, while the control group took the corresponding saphenous vein of the lower limb on the same side. Pathological tissue observation was performed using HE staining, Masson staining, and immunohistochemical examination for interleukin-2 (IL-2) and tissue inhibitor of metalloproteinases (TIMP-1). Result Out of the 22 rats in the surgical group, 20 were successfully modeled, with a success rate of 91%. According to the manifestations of venous dilation, varicose veins, and redness in the lower limbs of rats, compared with the control group, the varicose vein score in the model group increased significantly (P<0.01). After administering the therapeutic dose of Maizhiling, the varicose vein score in the Maizhiling group decreased significantly compared with the model group (P<0.01).Pathological examination showed significant varicose vein like changes and mild inflammation in the model group; The Maizhiling group reduced varicose veins and inflammation. Conclusion A rat model of lower limb varicose veins has been successfully established, providing a new research method for the study of drugs and treatment methods related to lower limb varicose diseases.
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Transcriptomics-based analysis of the effect of CompoundDancaoGranules on hepatocyte apoptosis in mice with non-alcoholic steatohepatitis

Li Xutao, Wang Siyuan, Ruan Tianyin, Zhang Hao, Peng Yuan, Liu Chenghai, Tao Yanyan

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Objective Transcriptome sequencing technology (RNA-seq) was used to analyze the mechanism of compound Dancao granules in the intervention of high-fat feed combined with carbon tetrachloride (CCl4) induced non-alcoholic steatohepatitis. Methods 45 male C57BL/6 mice were split into two groups at random: normal control group,model control group, obeccholic acid group (10mg/kg/d), Compound Dancao Granules low and high dose groups (10.71g/kg/d, 21.42g/kg/d), with 9 mice in each group. Normal diet was served to the control group, and the mice in the model group were given high-fat diet combined with subcutaneous injection of CC14, 100%CC14 solution (4mL/kg) for the first time, and 40% CC14-olive oil solution (2mL/kg) for the second time, twice a week for a total of 6 weeks. Each drug group was administered from week 3 for a total of 4 weeks. 12h after the last administration, serum and liver tissues of mice in each group were collected and biochemical kit was used to detect serum liver function. Hematoxylin-eosin (HE), Sirius scarlet, and oil red O stainings were used to examine the histopathological changes of the liver. Differential genes were analyzed by RNA-seq and performed functional enrichment analysis, in order to verify the expression of differential gene mRNA, the quantitative real-time polymerase chain reaction was used, and TDT-media atedDuTPnick-endlabeling (TUNEL) staining was used to identify the apoptosis. Results Model control groups had significantly higher levels of serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), total cholesterol(TC), and triglycerides(TG) than normal controls. (P < 0.01). Additionally, There was obvious inflammatory cell infiltration in liver tissue, collagen deposition in sink area and interlobule, and significant increase in lipid droplet area (P<0.01)..The levels of TC, TG, ALT, and AST were significantly lower in the groups receiving compound Dancao granules and obeccholic acid than in the model control group. (P < 0.01), inflammatory cell infiltration, collagen deposition and fat accumulation in the sink area and interlobule were improved (P<0.01). RNA-seq sequencing results showed that 2819 genes in the normal control group were differentially expressed compared with the model control group, 543 up-regulated genes and 2276 down-regulated genes. Between the model control group and the compound Dancao granules group, 240 genes were differentially expressed, including 206 up-regulated genes and 34 down-regulated genes. There were 221 overlapping genes in the two groups, and the functional enrichment mainly focused on cell cycle(CDT1, PLK1, BUB1B, TTK, KNL1, ESCO2, CDC6, NDC80, CDC25B, SGO1, CCNB2, ESPL1, CCNE1, MCM4, MCM5, FBXO5, BUB1, MCM2), apoptosis（Caspase-3，Bax，P53，Apaf1，Bak，Caspase-8）, P53 signaling pathway（P53, CCNB2, APAF1, BAK, BAX, GTSE1, CASPASE-3, CCNE1）, arachidonic acid metabolism（HPGDS, CYP2C54, CYP2B10, TBXAS1, CYP2C50）, galactose metabolism（HK3, GLA, HK2, AKR1B7） and other signaling pathways. RNA-seq sequencing analysis showed that compound Danicao granules mainly regulated apoptosis signaling pathway, and quantitative PCR confirmed that compared with normal control group, mRNA expressions of Caspase-3，Bax，P53，Apaf1，Bak and Caspase-8 in liver tissue of model control group were increased (P < 0.01). Compared with model control group, mRNA expressions of Caspase-3，Bax，P53，Apaf1，Bak and Caspase-8 in liver tissue of compound Dancao granules group were decreased (P < 0.01). TUNEL staining showed that the number of cell nuclear shrinkage and apoptotic bodies decreased in the group of compound Dancao granule administration. Conclusion Compound Dancao Granule has a significant protective effect on non-alcoholic steatohepatitis induced by high fat feed combined with CCl4, and its mechanism might be connected to the control of genes linked to apoptosis.
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Characterisation of cellular expression of GFP and pattern of response to diphtheria toxin in a Lyz2-IRES-DTREGFP mouse strain in vivo

yan

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【】Objective Observation of depletion of GFP-positive cells by diphtheria toxin in Lyz2-IRES-DTREGFP mice in vivo. Methods Transgenic mice of 6-8 weeks of age and wild-type mice of the same genetic background were randomly selected and subjected to intraperitoneal injection of 25 ng diphtheria toxin for three consecutive days, after which myeloid cell depletion was observed at different time points by flow cytometry (FCM). The combination of GFP and CD11b was used as a marker of myeloid cells (macrophages and neutrophils). Cells from bone marrow, peripheral blood and spleen were taken after intraperitoneal injection and subjected to FCM to determine the pattern of GFP-positive cells over time after diphtheria toxin injection. Results GFP-positive cells in peripheral blood decreased significantly on the third day after diphtheria toxin injection, and GFP-positive cells in spleen and bone marrow decreased significantly on the first day after injection, after which GFP-positive cells began to recover gradually. Conclusion Diphtheria toxin has a depleting effect on GFP-positive cells in Lyz2-IRES-DTREGFP mice in vivo, and the time of depletion varies at different sites.
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Effect of Shugan Huazheng-Fang powder on liver fibrosis in rats based on JAK2/STAT3 signaling pathway

Tian Meng-yuan, Chen guang-shun, Zhao Kun-peng, Zhang Qiu-ju, Zhang Ming

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Objective：To investigate the effects and mechanisms of Shugan Huazheng Decoction in rat liver fibrosis model and hepatic stellate cells (HSC-T6). Method：Fifty-four male SD rats were randomly divided into blank group, model group and low-dose (7.38g/kg), medium-dose (14.76g/kg) and high-dose (29.52g/kg) Shugan Huazheng Decoction group and colchicine group (0.2mg/kg), with 8 rats in each group. Rats in the model group and each dosing group were injected intraperitoneally with 40% carbon tetrachloride (CCl4) solution, while each dosing group was given the corresponding dose by gavage daily for 8 weeks. The liver serum biochemical indexes ALT and AST were detected by fully automatic biochemical analyzer; HE and Sirius staining was used to observe the liver tissue injury; ELISA detected the expression levels of α-SMA and Col Ⅰ in liver tissue; Western blot detected the protein expression levels of JAK2, P-JAK2, STAT3, P-STAT3, IL-6, IL-1β, and TNF-α; and RT-qPCR detected the mRNA expression levels of JAK2, STAT3, IL-6, IL-1β, and TNF-α in liver tissue. HSC-T6 cells were divided into blank group, model group, drug-containing serum group, inhibitor group and combined group, except blank group, all were induced by LPS, cell viability was detected by CCK-8; α-SMA, Col Ⅰ expression level was detected by ELISA; JAK2, P-JAK2, STAT3, P-STAT3, IL-6, IL-1β, TNF-α protein expression level in cells was detected by western blot JAK2, P-JAK2, STAT3, P-STAT3, IL-6, IL-1β, and TNF-α protein expression levels in cells; RT-qPCR detection of JAK2, STAT3, IL-6, IL-1β, and TNF-α mRNA expression levels in cells. Result：Shugan Huazheng Decoction dose-dependently improved liver function and attenuated hepatic fibrosis; the expression of JAK2/STAT3 signaling pathway and inflammation-related factors were significantly reduced in the positive control group and all Shugan Huazheng Decoction dose groups compared with the model group(P＜0.05)；Shugan Huazheng Decoction -containing serum significantly down-regulated the expression levels of α-SMA, Col Ⅰ, JAK2/STAT3 signaling pathway and inflammation-related factors in LPS-induced HSC-T6 cells(P＜0.05).Conclusion：Shugan Huazheng Decoction showed anti-hepatic fibrosis effects in rats in vivo and in vitro, and the mechanism of action was related to the expression of JAK2/STAT3 signaling pathway and inflammation.
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Establishment and exploration of a subcutaneous transplanted tumor model of hepatocellular carcinoma in mice with Qi stagnation and blood stasis syndrome

Cao Linna, Li Mingzhe, Shi Zhanhao, Wang Xiangxiang, Pan Zhiqiang, Peng Peike

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Objective: To explore the establishment of a subcutaneous transplanted tumor model of hepatocellular carcinoma in mice with Qi stagnation and blood stasis syndrome. Methods: Forty male C57BL/6 mice were randomly divided into 4 groups: NC group, QZXY group, Tumor group, and QZXY+Tumor group. They were categorized based on the modeling of Qi stagnation and blood stasis syndrome (7 days) combined with the modeling of subcutaneous transplantation tumor of hepatocellular carcinoma (20 days). Observations were conducted on syndrome manifestations, as well as tumor size and weight of the mice after modeling. Results: (1) Body weight: On the 7th day of modeling, the weight of QZXY group and QZXY+Tumor group was significantly lower than that of NC group (P<0.05). (2) Body temperature: On the 7th day of modeling, compared with NC group, the body temperature significantly decreased in QZXY group (P<0.05), while it increased in Tumor group (P<0.05). On the 27th day of modeling, the temperature of QZXY+Tumor group was significantly lower than that of NC group (P<0.05). (3) Syndrome manifestations: According to the syndrome scoring table, on the 7th day of modeling, mice in both QZXY group and QZXY+Tumor group exhibited Qi stagnation and blood stasis syndrome (P<0.05). As the modeling time extended, the score of mice in Tumor group increased with the formation of the tumor, the score of mice in QZXY+Tumor group was significantly higher than that of QZXY group (P<0.05). (4) Claw petechiae: Compared with NC group, the number of the claw petechiae was significantly increased in all three groups of modeled mice (P<0.05), with the QZXY+Tumor group showing the highest number. (5) Claw r value: Compared with NC group, the r value of the claw was significantly lower in all three groups of modeled mice (P<0.05). Additionally, the r value of the claw in the QZXY+Tumor group was consistently lower than that of the other three groups. (6) Open field activity: Compared with NC group, the vertical and horizontal activity of mice in QZXY+Tumor group decreased significantly (P<0.05). (7) Four coagulation indexes: Compared with NC group, APTT, TT and FIB were significantly increased in QZXY+Tumor group (P<0.05 or P<0.01). (8) Tumor size and weight: Compared with Tumor group, the tumor size and weight of QZXY+Tumor group increased significantly (P<0.05). Conclusion: This study successfully established a subcutaneous transplanted tumor model of hepatocellular carcinoma in mice with Qi stagnation and blood stasis syndrome. The findings indicate that Qi stagnation and blood stasis syndrome may occur during the course of liver cancer and could potentially further promote its development.
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Observation of Chinese medicine syndrome type and characterization of plasma metabolome in spontaneously hypertensive rats

Zhangqiuju, wangyu, lingbishi, simeilong, menghaoxian, tianmengyuan, zhangming, liannini, kangwanrong, jinhua

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OBJECTIVE To investigate the characteristics of Chinese medicine syndromes and possible metabolic substance basis of spontaneously hypertensive rat (SHR), a common model animal for hypertensive research. METHODS The general state, temperament, peripheral vascular filling, tongue image, diet and water intake, urine and feces volume and characteristics, blood pressure, heart rate, respiratory rate, pain threshold and open field of SHR rats were observed and tested comprehensively to identify the possible syndrome types of Chinese medicine. At the same time, Liquid Chromatography Tandem mass Spectrometry (LC-MS/MS) was used to analyze non-targeted serum metabolites, so as to preliminarily reveal the material basis of blood pressure elevation and Chinese medicine syndrome manifestations. RESULTS Compared with WKY group, SHR was dark yellow, irritable state and peripheral capillary filling score were significantly increased (P≤0.05); Red tongue color, dry tongue, little body fluid; Quantity of feces, urine, diet and drinking water decreased significantly, and the water content of feces was low (P≤0.05); Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), heart rate (HR) and respiratory rate (RR) were significantly increased(P≤0.05); Pain threshold decreased(P≤0.05); The open field experiment showed that the moving distance and residence time of the edge increased (P≤0.05). Serum non-targeted metabolomics results showed that compared with WKY group, there were 114 metabolites with significant differences in SHR (P≤0.05). These differential metabolites were mainly Lipids and lips-like molecules (40.35%), Organic acids and derivatives (22.8%), and Organoheterocyclic compounds (15.79%). A total of 25 metabolic pathways were obtained by KEGG enrichment analysis. Further differential abundance analysis showed that 16 pathways were activated, only 4 pathways were inhibited, and 5 pathways were not significantly changed. The Glutamatergic Synapse (Glutamatergic Synapse) and gamma-aminobutyric acid Synapse (GABAergic Synapse) were activated, while the 5-serotonergic synapse (Serotonergic synapse) was inhibited. CONCLUSION The symptoms of SHR include impatience and irritability, peripheral vascular dilation and collateral circulation formation, bulbar conjunctival congestive swelling, red tongue color, dry tongue, constipation, red yellow urine with little urine, rapid heart rate and respiratory rate, etc. All these suggest that SHR is a syndrome of hypertension with hyperactivity of liver-yang. The material basis of SHR is not only related to lipid and amino acid and carbohydrate metabolism disorders, but also may be related to the activation of excitatory neurometabolic pathway and inhibition of inhibitory neurometabolic pathway.
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Comparative research of flammatory factors in Differtent specimens of animal model of aspiration pneumonia in rats induced by lipopolysaccharide

songling, gaoyunhang, lihan, chentengfei, houhongping, yezuguang, zhangguangping

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To observe the change of Inflammatory factors in bronchoalveolar lavage fluid(BALF), lung tissue and serum of the rat pneumonia model induced by inhalation of lipopolysaccharide ( LPS ). Methods At 3 days after modeling with inhalation of LPS 4 mg/mL，15min/d, and monitoring the particle size distribution and aerosol concentration of LPS, inflammation degree of HE staining lung tissue of rats in each group was observed, and the neutrophils in BALF counted by microscope, the contents of interferon gamma(IFN-γ), interleukin-1bata(IL-1β), interleukin-4(IL-4), interleukin-5(IL-5), interleukin-6(IL-6), interleukin-10(IL-10), interleukin-13(IL- 13),tumor necrosls factor alpha(TNF-α), KC/GRO in lung tissue, serum and BALF were detected by Meso Scale Discovery( MSD) ．Results Histopathology in lung tissues of model rats behave focal and diffuse alveolar epithelial necrosis with shedding, and induced the aggregation and infiltration of inflammatory cells. The particle size distribution of atomized LPS as follows, Dv(10)=0.6974(μm), Dv(50)=3.387(μm) Dv(90)=8.836(μm). The aerosol concentration of LPS is 4.08g/m3, and the calculated inhalation dose for rats is 47.10 mg/kg. The contents of neutrophils count of model rats) increase significantly in BALF; The contents of IL-1β,IL-6 and KC/GRO in lung tissue of model rats increase significantly(P<0.01, P<0.05, P<0.01). The contents of IL-1β, IL-6 and TNF-α in BAIF of model rats increase significantly(P<0.05, P<0.001, P<0.001). No biological Significant changes was observed in Inflammatory factors of serum. Conclusion IL-1β、IL-6、KC/GRO and TNF-α changed significantly in model rats induced by inhalation of lipopolysaccharide ( LPS ). The model rats are local inflammation of the lungs, and inflammatory factors of serum had no significant changes. Inflammatory factors in lung tissue and lung lavage fluid are more prone to significant changes.
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Study on animal models of systemic lupus erythematosus

laiqin

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【】Systemic lupus erythematosus (SLE) models are mainly divided into four categories: spontaneous, induced, Humanized Mouse Models and gene knockout models. Lupus animal models are of great significance for the study of the etiology, pathogenesis and treatment of SLE. By reviewing and comparing various lupus animal models, this paper elucidates the main features, advantages and disadvantages of different types of SLE models, and provides a reference for researchers to select appropriate models in the process of exploring genetic and environmental factors, new pathogenic mechanisms and therapeutic modalities of systemic lupus erythematosus.
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Establishment and evaluation of qi yin deficiency syndrome model of type 2 diabetes in guizhou miniature pigs

qiuhuaming, lizixiang, tangjie, wuyanjun, zhaohai, wushuguang

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【】Objective The purpose of this paper is to explore the construction method, evaluation system and related indicators of the Qi Yin deficiency syndrome model of type 2 diabetes in Guizhou miniature pigs. Methods Twenty six six month old Guizhou miniature pigs were selected, of which six were fed with basic diet for 10 months in the control group, 20 were fed with high sugar and high fat diet for 10 months in the model group of qi yin deficiency in type 2 diabetes, and three months were fed with Qingpi Fuzi decoction. The model of qi yin deficiency in type 2 diabetes in Guizhou miniature pigs was constructed. Five scales were used to evaluate traditional Chinese medicine syndrome types, including urination, defecation, movement and mental state, response to the surrounding environment, and color of the skin, hair, nose, and disc. Serum CD4+, CD8+, and their ratios, cAMP, cGMP, and their ratios, as well as tongue R, G, and B values were used to evaluate Qi Yin deficiency syndrome. Fasting blood glucose (FPG), glycosylated hemoglobin (HbA1c), glucose tolerance test (OGTT), insulin resistance index (HOMA-IR), TG, TC, HDL-C, LDL-C, PT, APTT and other indicators were used to evaluate and evaluate type 2 diabetes. Results The physical signs of the small pigs in the model group can be summarized as overweight, fatigue and fatigue, tendency to lie down, laziness, lethargy, lack of energy, overeating and drinking, dull fur, and dull fur; The scores for defecation, movement and mental state, response to the surrounding environment, and color of the skin, fur, and nasal disc were relatively low (P<0.05); The tongue appears red with no or little coating; Compared with the normal group, the laboratory indicators of the small pigs in the model group showed FPG HbA1c、HOMA-IR、TC、LDL-C、cAMP、cGMP、 The R, G, and B values of tongue images were significantly reduced, while PT, APTT, CD4, and CD4/CD8 values were significantly reduced. TG was elevated (P>0.05), HDL-C was elevated (P>0.05), and cGMP was elevated (P>0.05); Pathological staining showed destruction of pancreatic tissue. Conclusions The method of using high sugar and high-fat feed combined with traditional Chinese medicine for breaking qi and damaging yin can successfully establish a small pig model of T2DM with qi and yin deficiency. The established model presents the main symptoms of fatigue and fatigue, thirst and thirst, shortness of breath and lazy speech, redness of the tongue and less fluid, lack of moss, and weak pulse in traditional Chinese medicine syndromes, as well as concurrent symptoms such as yellow and transparent urine, dry and dry stools, slow response, curling up, dull and dull fur, gray and dull nasal discs, as well as clinical manifestations of abnormal glucose and lipid metabolism and insulin resistance.
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Nuclear Factor κB Signaling Pathway in Viral Myocarditis

QiuYueqing, CHEN Zhenyi

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Nuclear factors κB (NF-κB) is a multifunctional transcription factor that participates in regulating a wide range of biological processes. Existing evidence suggests that the early pathological changes of viral myocarditis (VMC) are mainly virus induced myocardial cell damage, and the inflammatory response can effectively clear the virus; In the late stage, it is the indirect inflammatory damage caused by the virus activating the immune response. At this time, uncontrolled inflammation can damage the myocardium, leading to cardiac dysfunction. Therefore, the balance between pro-inflammatory and anti-inflammatory factors plays a decisive role in the prognosis of VMC. NF-κB pathway not only mediates inflammatory responses, but also regulates pathways such as cell apoptosis, energy metabolism, oxidative stress, and insulin resistance to participate in the pathological progression of VMC.
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Research progress of targeted CAF strategy in PDAC immunotherapy

LIU Xingyu, QIN Jing, HU Yaohua, GUO Mengtian, ZHAO Jumei, SHI Changhong

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【】 Pancreatic ductal adenocarcinoma (PDAC) is a common type of pancreatic cancer, which is insidious, rapidly development and highly malignant. Traditional treatment strategy are not effective for PDAC due to its rich extracellular matrix (ECM). Cancer associated fibroblasts (CAF) are the most important component of ECM. It can interact with other immune components in the tumor microenvironment (TME) by secreting a large number of effector molecules to form immunosuppressive TME, which could allow cancer cells to evade immune system surveillance, promote tumor growth, invasion and metastasis, induce ECM remodeling and drug resistance. This review provided a summary of the research progress on the application of targeted CAF in PDAC immunotherapy. It focus on exploring research strategies that promote the transition of TME from an immunosuppressive state to an immune activated state through depleting CAF, inhibiting effector molecules secreted by CAF, reprogramming CAF, and limiting CAF-induced ECM remodeling. The aim of this review is expected to produce more effective therapeutic strategies and provide new methods for the immunotherapy of PDAC
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Construction and phenotypes of Apoe gene knockout mice

KE Lu, CAO Yuan, GU Wenda, LIU Susu, SUN Xiaowei, ZHAO Haoyang, ZHAI Shijie, GUO Huaiyong, LOU Yue, FAN Changfa

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Objective: Apolipoprotein E (Apoe) plays a crucial role in lipid metabolism. The CRISPR/Cas9 was utilized to generate an Apoe knockout mouse model for further investigation the role of Apoe in lipid metabolism and atherosclerosis. Method: Two sgRNA designed for the Apoe in C57BL/6J mice were co-injected with Cas9 mRNA into fertilized eggs, followed by transplantation into ICR recipient mice to obtain F0 generation mice. Positive mice were identified by PCR screening of extracted tail DNA. The expression of Apoe mRNA in various tissues of mice was assessed by quantitative real-time PCR, while lipid indexes were measured in serum samples of mice. The lipid accumulation in the inner lining of aortic vessel was detected by Oil red O staining. Result: PCR and sequencing results confirmed the successful construction of Apoe knockout mice (C57BL/6-Apoeem1/Nifdc, referred to as Apoe KO); the results of quantitative real-time PCR exhibited that the expression of Apoe mRNA was significantly reduced in the liver, brain, spleen, kidney, and lung tissues of the Apoe KO homozygote mice (Apoe-/-). Additionally, the elevated serum total cholesterol and low-density lipoprotein cholesterol levels were occurred in Apoe-/- mice, and lower high-density lipoprotein cholesterol levels were also occurred in males. Compared to wild-type mice, the extensive lipid plaques in the inner lining of arteries were appeared in the Apoe-/- mice under normal chow consumption conditions. Conclusion: This study was successfully established an Apoe knockout mouse model exhibiting typical abnormal lipid metabolism phenotype with arterial lipid accumulation even without a high-fat diet intervention. This work provides background data for the Apoe KO mouse resource and a new model for the study of abnormal lipid metabolism phenotype.
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Research status and prospect of animal models of hearing loss

liuzexuan, daijianyu

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As a disease with complex pathogenic factors and unclear pathogenesis,hearing loss can cause low hearing inpatients,which seriously affects the health and quality of life of patients. In order to better explore the pathogenesis of hearing loss and provide a basis for its treatment research, it is of great value to establish an animal model that can simulate human hearing loss. The existing classical methods for modeling are drug injection, including intrabitoneal injection of cisplatin,gentaminic, mitochondrial toxin,D-galactose,furoxide and kanamycin, neomycin and congenital cytomegalovirus, etc. Physical modeling methods include noise, cervical spine injection of shardener, ischemia reperfusion, vasopressin injection, etc. In addition, there are other novel modeling methods such as genetic modification technology. In this paper, the advantages and disadvantages of the above-mentioned modeling methods are reviewed, hoping to provide a basis for further research on the modeling methods of hearing loss.
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Application of mink as a new experimental animal model

hejialei, hesong, zhangqing, zhangxiwen, zhanghe, yuanbao, chenjian, renwenzhi, hujinping, dingyu

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The mink, as a small and valuable fur-bearing animal, not only possesses significant economic value but has also gradually demonstrated immense potential as a novel experimental animal model due to its unique biological characteristics and similarities with humans in respiratory systems, immune responses, and other aspects. This paper comprehensively reviews the applied research on mink as an experimental animal model, encompassing its use in influenza virus animal infection models, COVID-19 animal models, animal behavior models, canine distemper animal models, vomiting models, enzyme digestion models, testicular degeneration models, and self-injurious behavior models. Additionally, the importance of mink animal welfare is emphasized, and the broad prospects for mink as an experimental animal model in scientific research are proposed, offering valuable insights and reference for the extensive application of mink as a novel experimental animal model in the future.
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Progress in the Application of Behavioral Evaluation Methods for Animal Models of Chronic Fatigue Syndrome in the Past Five Years

QU Yuanyuan, FENG Chuwen, SUN Weibo, SUN Zhongren, LI Binbin, LU Jing, SHAO Yuying, GUO Shuhao, CHEN Tao, YANG Tiansong

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Chronic fatigue syndrome (CFS) is a disease that has gradually increased in incidence in recent years, is easily overlooked in clinical diagnosis, and severely affects the quality of life of patients. Elucidating its pathogenesis and treating it from both etiology and symptomatology perspectives are crucial for the clinical recovery of CFS patients. However, due to the unclear pathogenesis, the etiological treatment options are limited, and current treatments mainly focus on improving clinical symptoms. In this context, establishing a good animal model and effectively evaluating it is particularly important. This article comprehensively integrates the diagnostic criteria of CFS and the progress of basic research, summarizes the behavioral experiment methods related to model evaluation involved in CFS basic research in the past five years, and discusses it for the first time from five aspects: general condition evaluation, fatigue state evaluation, cognitive function evaluation, emotional state evaluation, and pain degree evaluation. The purpose is to present the current situation, expose problems, trigger reflection, and promote improvement.
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Optimization of modeling method for experimental lung fibrosis induced by bleomycin in mice

ZHU Dongwei, ZHAO Qi, BAI Le, PAN Tingyu, WANG Jing, WEI Yun, XU Yong, ZHOU Xianmei

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Objective: To compare the effects of intratracheal instillation by lumbar spinal needle (the best modeling approach explored by our group in the early stage) and intratracheal atomization on bleomycin-induced pulmonary fibrosis model in mice, so as to find out the optimal modeling method. Methods: Seventy-two C57BL/6J mice were randomly divided into blank group, lumbar spinal needle group, and aerosolization group, according to body weight, with 24 in each group. Mice in the blank group were given intratracheal instillation of saline. Mice in the lumbar spinal needle group were instilled with bleomycin in trachea, and mice in the aerosolization group was aerosolized with bleomycin in trachea by microsprayer aerosolizer. On 14th day and 21st day, Micro-CT, histopathological changes, hydroxyproline level, collagen immunohistochemistry and α-SMA protein expression were examined to evaluate the degree of pulmonary fibrosis in each group. Results: Compared with the blank group, the mice in the two model groups showed listlessness and slow response. Compared with the blank group, the weight of mice in the two model groups decreased significantly on 14th and 21st days (P < 0.01). Micro-CT results showed that the white shadows in the lumbar spinal needle group surrounded the trachea, while those in the aerosol group were more diffuse. The degree of alveolitis and pulmonary fibrosis was the highest in the aerosolization group, with a time-dependent trend. The hydroxyproline content in the two model groups increased significantly on day 14 and day 21 after modeling (P<0.05). And the increase of hydroxyproline concentrations on day 21 was more significant and stable (P<0.001). The expression of Collagen I in the two model groups increased significantly after 21 days of modeling, especially in the aerosolization group. The results of α-SMA protein detection showed that the expression of α-SMA in the two model groups was significantly higher than that in the blank group after 21 days (P<0.01). However, there was no significant difference between the two model groups. Conclusion: Intratracheal atomization of bleomycin is an optimized scheme for the establishment of pulmonary fibrosis models.
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In vivo study on antibacterial carbon dots modified PEEK to improve osseointergration performance of implants in treating MRSA infection environment

HE Hongxing, ZHANG Xintian, ZHANG Menghan, WANG Yao, DENG Xiaoqin, WENG Shaohuang

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Purpose This study aims to evaluate the bone integration performance of antibacterial carbon dot modified polyether ether ketone (PEEK) in infectious bone defect environments Methods Guanidine based carbon dots (G-CDs) prepared by melting method combined with dialysis purification were applied to modify PEEK implants using polyvinyl butyraldehyde (PVB) by soaking-drying method (PEEK/PVB-G-CDs). The SD rats were divided into: (1) PEEK implanted uninfected group (PEEK (-) group), (2) PEEK/PVB-G-CDs implanted uninfected group (PEEK/PVB-G-CDs (-) group), (3) PEEK implanted infected group (PEEK (+) group), (4) PEEK/PVB-G-CDs implanted infected group (PEEK/PVB-G-CDs (+) group). A hole with a diameter of 2 mm and a depth of 5 mm at the lateral condyle of the vertical femur were drilled on each group of rats to simulate a bone defect. The PEEK (-) and PEEK/PVB-G-CDs (-) groups without infection were injected 30 μL physiological saline into the bone marrow cavity of each rat. The PEEK (+) and PEEK/PVB-G-CDs (+) groups with infection were injected 30 μL MRSA bacterial suspension (1.5 × 104CFU/mL) into the bone marrow cavity of each rat. At 0, 2, and 4 weeks after implantation, animal specific X-ray was used to observe the implantation site of each group of rats. After 6 weeks of surgery, Micro CT was used to evaluate the bone tissue characteristics of the implantation site. Bacterial culture of bone marrow, Hematoxylin eosin staining, Toluidine blue staining, Goldner trichrome staining, and immunohistochemical staining were used to analyze the bone implantation sites of each group of rats. Results (1) X-ray, Micro CT, bacterial culture of bone marrow, and histopathological analysis confirmed that there was no infection in the PEEK (-) and PEEK/PVB-G-CDs (-) groups. The implants were integrated with bone defects. The PEEK/PVB-G-CDs (+) group exerted antibacterial activity to effectively control osteomyelitis caused by MRSA and achieved bone integration, while the PEEK (+) group was unable to achieve bone integration due to persistent infection. (2) Immunohistochemical staining evaluation confirmed that the PEEK (+) group showed lower levels of anti-inflammatory factors such as IL-4 and IL-10, as well as stronger expression of pro-inflammatory factors such as IL-6 and TNF-α compared to the other three groups, indicating that G-CDs modified PEEK can inhibit MRSA infection, regulate the inflammatory level of the local microenvironment, and promote bone integration at the site of bone defects. Conclusions Antibacterial carbon dots modified PEEK exhibits excellent bone integration performance, providing a candidate strategy for future clinical treatment.
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Macrogenomics-based investigation of the mechanism of action of GeGen QinLian Decoction in ameliorating dysbiosis in an SD rat model of antibiotic-associated diarrhea

tang qiumei, han xue, yang guangyong, chen rui, wang wenjia, tu xiaohua, tian weiyi, Cai yanpeng, Chen chi, He guangzhi

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Abstract：Objective: To investigate the changes of intestinal flora as well as function after treatment of rats with antibiotic-associated diarrhea with GeGen QinLian Decoction. Methods: Sixty SPF-grade male and female SD rats were fed for 7 days and randomly divided into blank and modeling groups according to 1:5. Rats in the modeling group were modeled by gavage with clindamycin 250 mg/kg once a day for 7 consecutive days. After successful modeling, the rats were randomly divided into the model group (Mod), the high dose group of GeGen QinLian Decoction (GQD-H), the medium dose group of GeGen QinLian Decoction (GQD-M), the low dose group of GeGen QinLian Decoction (GQD-L), and the Live Bifidobacterium Power group (LBP), with 10 rats in each group. According to the administered doses of GQD-H (10.08 g/kg), GQD-M (5.04 g/kg), GQD-L (2.52 g/kg), and LBP (0.15 g/kg) by gavage, the blank and model groups were given an equal volume of saline by gavage once a day, and feces were collected after 7 consecutive days of administration for macro-genomics sequencing analysis. Results: α-diversity and β-diversity suggested that there was a difference in intestinal microbial diversity between the Mod group and the GQD-treated group; GQD increased the abundance of the phylum Thick-walled Bacteria and decreased the abundance of the phylum Aspergillus at the phylum level. Increased relative abundance of intestinal mucus fungi Blautia, Bacteroides, Thomasclavelia, Mediterraneibacter and decreased relative abundance of Adlercreutzia, Muribaculum, Escherichia at the genus level; GQD up-regulated amino acid metabolic pathway, carbohydrate metabolic pathway, and immune disease pathway. Conclusions: GQD intervention in AAD improves the abundance ratio of beneficial and pathogenic intestinal bacteria, which in turn reduces the intestinal inflammatory response and repairs the intestinal immune system.
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The effect of bear bile powder and ursodeoxycholic acid on preneoplastic lesions of hepatocarcinoma

JIA Guiyang, NI Yuanping, WANG Mengru, MAKOTO Shibutaniand JIN Meilan, jinmeilan

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Objective To investigate the effect of bear bile powder (BBP) and ursodeoxycholic acid (UDCA) on preneoplastic lesions of hepatocarcinoma, we conducted anticancer efficacy experiments of BBP and UDCA using short-term carcinogenesis bioassay models. Methods Forty 6-week-old male SD rats were randomly divided into control, DEN, DEN+BBP (200 mg/kg), and DEN+UDCA (30 mg/kg) groups. At the beginning of the experiment, except for the control group, all rats were injected intraperitoneally with 100 mg/kg DEN once a week for three weeks. Additionally, all rats of the last two groups were orally administered 200 mg/kg BBP or 30 mg/kg UDCA suspended solution daily from the beginning to the end of the experiment. Results In the body and liver weights, there were no significant differences between DEN, DEN+BBP and DEN+UDCA groups. The accumulation of MDA was increased by DEN treatment while the activities of SOD, GSH were decreased in the liver tissue. However, the activities of SOD, GSH were enhanced with UDCA administration, while the accumulations of MDA was decreased in the liver tissue. Contrary, BBP exerted these antioxidant effects in serum. Meanwhile, the number and area of GST-P positive lesions and Ki-67 positive cell ratio in the DEN+BBP and DEN+UDCA groups were significantly lower than those in the DEN group, and more obvious in DEN+UDCA group. In addition, the mRNA expression level of Caespase-9 was significantly increased by UDCA treatment than that of the model group. Conclusion These results suggested that BBP and UDCA have significant inhibitory effects on preneoplastic lesions of hepatocarcinoma induced by DEN, and both have antioxidant effects on DEN-induced oxidative stress. However, their antioxidant mechanisms are different. In addition, further research is needed to determine whether the antioxidant effect is involved in their anticancer mechanisms.
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Histological evaluation and intestinal flora analysis of rat model of colitis-cancer transformation induced by AOM/DSS

WEI Xiunan, SUN Dajuan, LIANG Junwei, LIU Jiahui, LI Gongyi, CHI Lili

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Objective The aim of this study is to establish a rat model of inflammation-cancer transformation of inflammatory bowel disease (IBD) and explore its intestinal flora characteristics. Methods Adult male Wistar rats were randomly divided into control group and model groups (M1, M2, M3) with different DSS intervention cycles. Rats in the model group were all induced colitis-cancer transformation by single intraperitoneal injection of AOM combined with free drinking of DSS in different cycles, during which DAI scores were recorded. Rats in M1, M2 and M3 groups were killed at the end of the first, second and third cycles of DSS, and spleen, colon tissue and colon contents were collected. Hematoxylin and eosin (H E) staining and transmission electron microscope (TEM) were used to evaluate the histological damage and carcinogenesis of colon in each group, and the characteristic changes of intestinal flora were analyzed by 16S rRNA sequencing technology. Results AOM/DSS resulted in significant increase of DAI score, shortening of colon and increase of spleen index. From M1 to M3 group, the intestinal mucosal barrier was gradually destroyed, and the pathological score was gradually increased. Abnormal crypt focus, polyp, low-grade intraepithelial neoplasia, high-grade intraepithelial neoplasia and mucosal carcinoma appeared in turn. At the same time, the pathological evolution process shows similar characteristics to the carcinogenesis of human inflammatory bowel disease. In 16S rRNA sequencing, two differential abundance testing tools, Wilcoxon and ALDEx2, were used to screen out that the changes of flora abundance represented by Bacteroidetes and Monoglobus may be involved in the progress of colitis-cancer transformation, and the functions of differential flora were mainly enriched in metabolic pathways such as lipid metabolism and carbohydrate metabolism. Conclusions The rat model induced by AOM/DSS can dynamically simulate the pathological characteristics of colitis-cancer transformation, accompanied by changes in the abundance of specific intestinal flora, which may be closely related to the metabolic pathway mediated by flora.
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Research Progress on the Role of PADI2 in Autoimmune Diseases

zhengzemin, liuke, zhangyanying, zhaoya, shichanghong

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Peptidyl Arginine Deiminase 2 (PADI2) is an enzyme that catalyzes the conversion of arginine residues to citrulline on protein peptides. Aberrant activation of PADI2 can induce excessive tissue inflammation and immune responses, thereby exacerbating the progression of autoimmune diseases (ADs). Through citrullination, PADI2 modifies protein structure and immunogenicity, influencing the production of autoantibodies and regulating the activity of immune cells such as neutrophils and macrophages. This review provides an overview of PADI2's functions and its pivotal role in ADs, with the objective of elucidating the mechanisms underlying ADs pathogenesis and identifying novel therapeutic targets and strategies for related diseases.
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Establishment and optimization of “atopic dermatitis-asthma” mouse model based on allergic march

Ren Meng Jiao, Yan Yi Ting, Zhang Yu, Qu Min Ye, Liu Tao

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Objective To establish and optimize the animal model of atopic march (AM), the skin and lung tissue sensitization under conditions of single or combined modeling of OVA and MC903 were compared. Methods 40 SPF BALB/c mice aged 6~8 weeks were randomly divided into control group, model group A, model group B and model group C. The three AM models were established with MC903, MC903+OVA and OVA, respectively, through twice skin sensitization and once respiratory sensitization. Skin sensitization severity scoring system, immunohistochemistry and enzyme-linked adsorption assay were used to compare skin lesion morphology, skin or lung histopathology and immunophenotype. Results Compared to OVA or MC903 modeling alone, MC903+OVA modeling mice showed more significant changes in skin morphology, higher score of skin sensitization severity, more severe skin and airway inflammatory cell infiltration, and more significant changes in the expression of related inflammatory factors TSLP, IL-4, IL-13 and IL-10 (all P <0.05). Conclusion The AM animal model optimized by MC903 combined with OVA was successfully constructed, which provided a good methodological basis for AM mechanism research.
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Research on the mechanism of oxidative stress in ovarian dysfunction

LIYUAN, ZHANGYISHA, YOUDIAN, TONGKAIWEN, XIANGYINGYING, CHENYICHEN, PEIYIWEN, ZHANGTING

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Age-related ovarian aging is significantly influenced by oxidative stress (OS) , which has intricate underlying mechanisms. Studies conducted recently have demonstrated that oxidative stress is a mediator of several pathological processes that lead to ovarian dysfunction in aging. These processes include telomere shortening, chronic inflammation, apoptosis, and mitochondrial dysfunction. Under oxidative stress, antioxidant treatment can assist in enhancing?ovarian function.
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Research progress of animal model construction of steroid- induced osteonecrosis of the femoral head

LEI Yang, LI Huiying, MENG Dongfang, ZHANG Xiangbei, WANG Kai

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As a complex and refractory disease, the incidence of steroid-induced osteonecrosis of the femoral head (SONFH) is increasing year by year and showing a younger trend. A good animal model of disease can provide important support for the study of the pathogenesis of SONFH and the development of treatment methods. In this paper, the latest progress in the construction of SONFH animal models was summarized and analyzed from the aspects of animal selection, modeling methods and model evaluation by reviewing and sorting out the experimental studies related to SONFH animal models at home and abroad in recent years, so as to provide reference for the related research of SONFH.
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Progress in Genetically Modified Animal Models of Hyperuricemia

LU Yongxin, LI Jia, LIU Mingyu, TAN Wenbin

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Gene knockout technology is increasingly becoming a powerful tool for establishing animal models of hyperuricemia (HUA). HUA gene knockout animal models are not only helpful to reveal the molecular mechanism of uric acid metabolism, but also of great value for evaluating potential therapeutic strategies. In this paper, the application of gene knockout technology in HUA animal models was discussed in detail by reviewing the domestic and foreign literature, focusing on urate oxidase (UOX), glucose transporter 9 (GLUT9) and ATP-binding cassette transporter G2 (ABCG2) were knocked out in experimental animals in order to provide reference and guidance for the further establishment of HUA animal models by gene knockout technology.
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Monoamine oxidase A: new target for tumor immunity from neural origin

ma yifan, li hui, chen hanmu, SHI Changhong

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Monoamine oxidase A (MAOA) is a membrane-bound mitochondrial enzyme existing in almost all vertebrate tissues that catalyzes the degradation of biogenic and dietary -derived monoamines. MAOA regulates neurotransmitter metabolism and is associated with anti-tumor immune responses. Previous most studies focused on the role of MAOA in tumor cells, while more recent findings suggest that MAOA plays an equally significant role in tumor-associated immune cells. In this review, we summarize the regulatory effect of MAOA on inhibitory tumor microenvironment, and the suppression function on the activity of various types of tumor-associated immune cells (e.g., CD8+T cells and tumor-associated macrophages) by directly affecting monoamines. We propose that developing novel MAOA-inhibitor drugs and exploring multidrug-combination strategies may enhance the efficacy of immune governance. Thus, MAOA may act as a novel target for tumor immunity and influence the effect of tumor immunotherapy
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Analysis of dilated cardiomyopathy animal models based on clinical characteristics of traditional Chinese and western medicine

TAO Shiyi, YU Lintong, Li Jun, YANG Deshuang, WU Jiayun, HUANG Li, Shao Mingjing, Huang Xuanchun

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Dilated cardiomyopathy (DCM) is one of the common diseases leading to heart failure, arrhythmia and sudden death. The etiology of DCM is complex and diverse, the mechanism has not been fully elucidated, and the intervention has limited improvement in the prognosis of patients, with a 10-year survival rate of less than 25%. This study aimed to summarize the construction characteristics of DCM animal model and evaluated the clinical compatibility of the model with traditional Chinese and western medicine based on domestic and foreign research results of DCM animal model, Western clinical diagnostic criteria and TCM syndrome differentiation. It was found that DCM modeling methods mainly involved gene editing, drug induction, immune induction, viral infection, rapid pacing induction. Experimental animals mainly included muroid, zebrafish, drosophila, pigs, thereinto, mice and rats were most commonly used. Gene editing was the most commonly used animal model of DCM, followed by doxorubicin-induced model. The literature review showed that experimental animals, drugs, single or cumulative doses, administration methods, and modeling period varied in different studies about the same DCM animal model. The level of clinical anastomosis of traditional Chinese and western medicine varied considerably, and the clinical anastomosis of traditional Chinese medicine was generally lower than that of Western medicine in the same model. In addition, the modeling standards of DCM animal models were mostly based on Western medicine theories, and the differentiation standards of syndrome models and the information collection standards of the four diagnoses have not been standardized and unified. In the future, stable and homogeneous animal models with high clinical consistency combining of disease and syndrome need to be established to provide a basis for DCM mechanism research and new drug development.
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Research progress of natural products in the treatment of hepatic fibrosis by regulating TGF-β1/Smad pathway

LI Kaiyang

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TGF-β1 is considered to be a key mediator in the formation of hepatic fibrosis, mainly by activating its downstream Smad signaling pathway. Smad2 and Smad3 are two major downstream regulators that promote TGF-β1-mediated tissue fibrosis, while Smad7 is a negative feedback regulator of the TGF-β1/Smad pathway and inhibits TGF-β1-mediated hepatic fibrosis. A growing number of studies have shown that natural products can delay the progression of HF by regulating the TGF-β1/Smad pathway, inhibiting HSC activation and reducing ECM deposition. This article reviews the molecular mechanism of TGF-β1/Smad signaling pathway in hepatic fibrosis, and summarizes the natural products that target the regulation of TGF-β1/Smad signaling pathway, in order to provide reference for the treatment of hepatic fibrosis.
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Research Progress on Rodent Models for Type 2 Diabetes Mellitus and Peri-implantitis

YAN Tingting, LI Hao

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Type 2 diabetes mellitus is a common metabolic disease characterized by hyperglycaemia and insulin resistance. Peri-implantitis is a common complication of oral implant. Type 2 diabetes mellitus can enhance bone destruction in peri-implantitis. However, the mechanism of association between the two has not been fully elucidated, and it is necessary to establish an effective animal model of type 2 diabetes mellitus complicated with peri-implantitis. At present, several methods have been proposed to construct rodent models of type 2 diabetes mellitus combined with peri-implantitis. This article reviews the establishment of rodent models of type 2 diabetes mellitus complicated with peri-implantitis. It can provide reference for the prevention, treatments and the establishments of animal models of type 2 diabetes patients with peri-implantitis.
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Inhibitory effect of Sauchinone on renal fibrosis in UUO mice and its mechanism

WU Pengyang, ZHAO Ling, LI Qiuyue, GUO Hongyan, LI Chaojun, WU Iina, zhaozhenglin

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Objective To investigate the inhibitory effect of Sauchinone (Sch) on renal tissue fibrosis and its mechanism in unilateral ureteral obstruction (UUO) mice. Methods Forty male ICR mice were taken and divided into sham-operated group (Sham group), UUO fibrosis model group (UUO-Model group), Sch low-dose group (Sch-Low group), Sch high-dose group (Sch-High group), and valsartan-positive control group (Val group). Starting on day 2 after surgery, the Sch low-dose group (10 mg/kg), the Sch high-dose group (30 mg/kg), and the valsartan-positive control group (100 mg/kg) were administered by gavage once daily for 4 weeks, respectively. At the end of drug administration, serum levels of creatinine (SCr), blood urea nitrogen (BUN), and inflammatory factors TNF-α and IL-6 were measured; SOD activity, MDA and ROS content were detected in renal tissues; the levels of inflammatory factors TNF-α and IL-6 mRNA were detected in renal tissues by RT-qPCR; HE, Masson staining, and immunohistochemistry were used to detect pathological changes and collagen deposition as well as TGF-β1, Smad3, and CTGF protein expression in mouse renal tissues; Western blot method was used to observe Collagen Ⅰ, CTGF, Smad3, Nrf2, HO-1 and NF-κB protein expression in renal tissues. Results Compared with the UUO model group, the serum levels of SCr and BUN as well as TNF-α and IL-6 were significantly reduced in the Sch low-dose and Sch high-dose groups of mice (P<0.05 or P<0.01); SOD activity was significantly higher and MDA and ROS levels were significantly lower in renal tissues (P<0.05 or P<0.01); The expression of TNF-α and IL-6 mRNA in renal tissues was significantly reduced (P<0.05 or P<0.01); Microscopic observation of swollen renal tubules with thylakoid hyperplasia and collagen fiber deposition was significantly improved; Immunohistochemistry showed a significant reduction in TGF-β1, Smad3 and CTGF protein expression in renal tissues; Western blot showed that Collagen Ⅰ, CTGF, Smad3, NF-κB protein expression was significantly reduced (P<0.05 or P<0.01); meanwhile, Nrf2, HO-1 protein expression was significantly elevated (P<0.05 or P<0.01). Conclusion The mechanism of the amelioration of renal tissue fibrosis by triphalaenone may be mediated through anti-inflammatory, anti-oxidative stress effects that modulate the expression of pro-fibrotic proteins of the TGF-β1/Smad3 signaling pathway.
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A preliminary study on the construction of low birth weight mouse Model based on Abi3bp gene knockout

Huang Yanqiu, Zhang Yue, Shi Liuliu, Zhao Xiaoying, Tang Junming, WU Yan

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Objective To employ the Abi3bp gene deletion mouse model for the detection of postnatal changes in body weight and glucose metabolism. Additionally, It also suggests a different method for low birth weight mice models. Methods A mouse model with the Abi3bp gene deletion was developed using CRISPR/Cas9 technology, followed by breeding and identification processes. To produce Abi3bp gene knockout homozygotes (Abi3bp-/-), heterozygotes (Abi3bp+/-), and wild type (WT) mice, heterozygotes were mated. Adult mice from all three groups were evaluated for glucose metabolism markers such as fasting blood glucose levels, glucose tolerance, and insulin tolerance, in addition to body weight at various postnatal time periods including the weight ratio of critical organs in adulthood. Results Gene identification confirmed successful knockout of the Abi3bp gene in Abi3bp-/- mice, with RT-qPCR analysis demonstrating significantly reduced Abi3bp expression in Abi3bp-/- mice compared to WT mice. Notably, the birth weight of Abi3bp-/- mice (1.25±0.08g) was markedly lower than that of WT mice (1.34±0.12g) (P<0.05). Conversely, the weight of adult (120d) Abi3bp-/- mice (27.70±1.93g) was significantly higher than that of WT mice (23.64±1.34g) (P<0.01). The ratio of key organs to body weight did not exhibit significant differences between the groups (P>0.05). Fasting blood glucose and insulin tolerance tests showed no significant variations between the groups. However, glucose tolerance tests indicated that Abi3bp-/- animals displayed lower blood glucose levels (15.68±7.04mmol/L) compared to WT mice（23.01±5.75 mmol/L）. Conclusion Deletion of the Abi3bp gene results in mice with low birth weight, growth recuperation, and inadequate glucose tolerance in adulthood similar to clinical growth traits of low birth weight neonates, thus presenting a promising choice for low birth weight mouse models.
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Effects of aqueous extract of Curcuma kwangsiensis root tubers on isoproterenol-induced cardiac hyPertroPhy in mice based on the sGC-cGMP-PKG signalling Pathway

He Yongyun, Li Hong, Hu Mingxia, Yao Yue, Mo Xiaodan, Yang Xiufen

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【Abstract】 Objective To investigate the role of sGC-cGMP-PKG signalling Pathway in the amelioration of isoproterenol (ISO)-induced cardiac hypertrophy in mice by aqueous extract of Curcuma kwangsiensis root tubers (GYJS) and the related mechanism. Methods The study involved seventy-two KM male mice, which were randomly divided into six groups: a normal control group, a model group, a propranolol positive control group (40 mg/kg), and three GYJS groups with low (1 g/kg), medium (2 g/kg), and high (4 g/kg) doses. Apart from the control group, mice in each group were subcutaneously injected with ISO (10 mg/kg) on days 1-3 and ISO (5 mg/kg) on days 4-14 to create a mouse cardiac hypertrophy model. After daily subcutaneous injections for 4 hours, the mice in each group were orally administered the corresponding drugs for a dosing cycle of 14 days. The weight of the whole heart and left ventricle in mice was measured. The pathology of mouse myocardial tissues was observed using HE and Masson staining. The expression of GUCY1B3 and TGF-β1 in myocardial tissues was observed using IHC staining. The kit was used to measure serum levels of LDH, CK, and NO, as well as the activity of SOD and the content of MDA in myocardial tissues. Serum levels of cGMP were measured using ELISA, and qPCR was used to determine the content of cGMP. qPCR was also used to determine the content of ANP, BNP, GUCY1B3, PKGⅠ, and PDE5A mRNA expression levels. It is important to adhere strictly to metrics and units when reporting results. Compared to the model group, the propranolol group and each dose group of GYJS significantly reduced the whole heart weight index and left ventricular index of mice (P<0.001 or P<0.0001). They also reversed the hypertrophy of myocardial tissues and myocardial fibrosis of mice and elevated the expression of GUCY1B3 in the myocardial tissues of mice (P<0.05 or P<0.001). The study found that treatment with compound 01) resulted in a significant reduction of TGF-β1 expression (P<0.05 or P<0.01). Additionally, it led to a significant decrease in myocardial injury markers LDH and CK activity (P<0.05 or P<0.01), a significant increase in NO and cGMP content (P<0.05 or P<0.01), a significant decrease in myocardial oxidative stress indicator MDA content (P<0.05 or P<0.01), and a significant increase in SOD activity (P<0.05 or P<0.01).Results Compared with the model group, the propranolol group and each dose group of GYJS could significantly reduce the whole heart weight index and left ventricular index of mice (P<0.001 or P<0.0001), significantly reverse the hypertrophy of myocardial tissues and myocardial fibrosis in mice, significantly elevate the expression of GUCY1B3 in the myocardial tissues of mice (P<0.05 or P<0.01), significantly reduce the expression of TGF-β1 (P <0.05 or P<0.01), myocardial damage markers LDH and CK viability were significantly reduced (P<0.05 or P<0.01), NO and cGMP contents were significantly elevated (P<0.05 or P<0.01), myocardial oxidative stress indicator MDA content was significantly reduced (P<0.05 or P<0.01), SOD viability was significantly increased (P<0.05 or P< 0.01), myocardial hypertrophy markers ANP, BNP and PDEA mRNA expression levels were significantly reduced (P<0.05, P<0.01 or P<0.001), and the mRNA expression of GUCY1B3 and PKG Ⅰ was significantly increased (P<0.01 or P<0.001).Conclusion GYJS may improve cardiac hypertrophy by modulating the sGC-cGMP-PKG signaling pathway.
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A study on the differences in emotional behavior between males and women in mice with Maoa c.1409 T>C synonymous mutants

Likaixin, liuguanglin, Yuanqianqian, Liufanglin, Chenhuan, Houhongwei

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Objective: To investigate the effect of monoamine oxidase A(MAOA)MAOA c.1409 T>C synonymous mutation on anxiety, fear and other emotional behaviors in mice. Methods: In this study, CRISPR/Cas9 technology was used to construct a mouse model of SNP synonymous mutation, which simulated the C and T allele polymorphisms of rs11370770 in the population, and evaluated the differential effect of this gene on male and female differences through animal behavior and gene expression levels in animal models. In terms of animal behavior, open field test (OFT), elevated plus maze (EPM), defensive burial experiment (DBE), and 3D behavioral analysis were mainly used analysis) and other methods were used to evaluate the behavioral differences between male and female males caused by polymorphisms in Maoa synonymous mutant genes, and the MAOA content and enzyme activity in mice were detected to evaluate the effect of Maoa rs1137070 polymorphisms on gene expression in male and female SNP mice. Results: The results of the open field experiment showed that the residence time of female SNP mice in the central area of the open field was significantly higher than that of male SNP mice (P<0.001), the results of elevated cross maze experiment showed that the time and frequency of male SNP mice entering the open arm were higher than those of female SNP mice, but there was no significant difference, and the results of defensive burial test showed that the number and duration of excavation of female SNP mice were significantly reduced to the threat of rat urine (P<0.01). Three-dimensional fine behavior analysis showed that there was no significant male and female difference except for the movement trajectory and climbing behavior of mice. Compared with male SNP mice, the MAOA enzyme content of female SNP mice was significantly lower than that of male SNP mice (P<0.001), but there was no significant difference in enzyme activity between male and female SNP mice. Conclusion: The synonymous mutation of MAOA c.1409 T>C may cause fear, anxiety and mood differences in male and female mice by affecting the expression of MAOA in male and female SNP mice.
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Protective effect and mechanism of Qingerkang Tea on apoptosis of mice cells with acute liver injury induced by carbon tetrachloride

TAN Dianbo, LIU Zhonghua, Du qing, Zeng Hongliang, Li Yuehui

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Objective Based on silencing information regulatory factor 1 (SIRT1)/high mobility group protein B1 (HMGB1)/nuclear transcription factor-κB (NF-κB) to investigate the inhibitory effect and mechanism of Qingerkang tea on hepatocyte inflammatory apoptosis in mice with carbon tetrachloride (CCl4) induced acute liver injury. Methods C57BL/6 mice were randomly divided into normal group, model group, SIRT1 agonist (resveratrol) group, Qingerkang tea low-dose, medium-dose and high-dose groups, and positive drug (dicycloalcohol tablets) group. The mice were given continuous intragastric administration for 14 days. Acute liver injury model was established by intraperitoneal injection of 0.5% CCl4 olive oil solution (5 mL/kg). The levels of alanine transferase (ALT), aspartate transferase (AST), lactate dehydrogenase (LDH) in serum and hydroxyproline (Hyp), malondialdehyde (MDA) and superoxide dismutase (SOD) in liver were determined by biochemical method. Serum levels of inflammatory tumor necrosis factor (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) were determined by enzyme-linked immunosorbent assay (Elisa). Eosin-hematoxylin (HE) staining and TUNEL staining were used to detect the pathological morphology and apoptosis of liver tissues. The protein expressions of SIRT1, HMGB1 and NF-κB were detected by Western blotting. Results Compared with the normal group, the serum ALT, AST, LDH levels and liver tissue Hyp activity in the model group were significantly increased, the activities of MDA and SOD in liver tissue were significantly decreased, the levels of inflammatory factors TNF-α, IL-6 and IL-1β in the liver tissue were significantly increased, and there were obvious pathological injury and hepatocyte apoptosis in the liver tissue. The expression of SIRT1 protein in liver tissue decreased significantly, while the expression of HMGB1 and NF-κB protein increased. Compared with model group, the serum levels of ALT, AST, LDH and Hyp activity of liver tissue were significantly decreased, the activities of MDA and SOD in liver tissue were significantly increased, the levels of serum inflammatory factors TNF-α, IL-6 and IL-1β were decreased, the pathological injury of liver tissue and the apoptosis of liver cells were significantly improved. The expression of SIRT1 protein in liver tissue was increased, while the expression of HMGB1 and NF-κB protein were decreased in Qingerkang tea high-dose group and resveratrol group. Conclusion Qingerkang tea could effectively protect acute liver injury, and its mechanism may be related to regulating SIRT1/ HMGB1/ NF-κB signaling pathway and alleviating hepatocyte inflammatory apoptosis.
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Establishing and Evaluating of Asthma Rat Model with Phlegm and Blood Stasis Syndrome

Yuan Xin, Wang Anqi, Wang Siyu, Pan Lexin, Wang Jiaying, Zou Lu, Yang Aidong

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Objective To explore the way to establish and evaluate the asthma rat model with phlegm and blood stasis syndrome. Methods 60 SD male rats were randomly divided into 5 groups:normal group,asthma model group,combination of disease and syndrome group(combination group),dexamethasone group,Kechuan Liuwei Mixture group),with 12 rats in each group. Asthma models were established by inducing ovalbumin (OVA) . Syndrome model of phlegm and blood stasis was established using a high-fat diet combined with ice water bath method. Evaluate asthma model through animal behavior observation,pathological section observation,inflammation index detection,and airway reactivity measurement; Evaluate phlegm and blood stasis syndrome model by measuring rat body mass,observing symptoms,detecting blood glucose,blood lipids,coagulation function,and hemorheological indexes,as well as syndrome determination by Kechuan Liuwei Mixture. Results ① After OVA inducing,the rats in the asthma model group and combination group showed symptoms such as shortness of breath,open mouth breathing,abdominal movement,restlessness,and irritability; HE staining showed disordered arrangement of the bronchial mucosa in lung tissue,local detachment,thickening of the basement membrane and the bronchial tube wall,narrowing of the lumen,extensive infiltration of inflammatory cells,and congestion of capillaries; compared with the normal group, IL-4,IL-6,TGF- β 1 in serum increased in rats of the asthma model group and combination group (P<0.05); Penh values increased after stimulation with various concentrations of Mch (P<0.05) .②Rats in the combination group showed symptoms such as chills,curling up with little movement,purple and dark claws,purple and black bruises on the tail,loose stools,and unclean perianal area; compared with the asthma model group,the body mass of rats in the combination group increased (P<0.05); blood glucose,triglycerides,and total cholesterol levels increased (P<0.05),thrombin time shortened (P<0.05),fibrinogen content increased (P<0.05),and whole blood viscosity significantly increased at low,medium,and high shear rates (P<0.05); the indexes were significantly improved after Kechuan Liuwei Mixture treating. Conclusion The asthma rat model with phlegm and blood stasis syndrome can be established through OVA inducing and high-fat diet combined with ice water bath. The model can be evaluated through behavioral observation,indexes detection,and syndrome determination by formulas.
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A superior method for modelling primary liver cancer in rats

zhangriyun, wufenglan, maodewen, wangminggang, peihao, lifeiyan

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Abstract：Objective： A rat model of primary hepatocellular carcinoma (HCC) was established using three different doses to establish an efficient, stable, and economical animal model of HCC. Methods： Forty-five male SD rats were randomly divided into four groups, namely the normal group, the diethylnitrosamine (DEN) 50 mg/kg dose group (low dose group), the 70 mg/kg dose group (medium dose group), the 200 mg/kg dose group (high dose group), 6 in the normal group and 13 in each of the remaining groups. The normal control group received no treatment. In the model group, the low dose group was injected intraperitoneally twice a week during weeks 1-4 and once a week during weeks 5-12; the intermediate dose group was injected intraperitoneally once a week for 16 consecutive weeks; and the high dose group was administered once in a single dose and the rats in each group were followed up to 16 weeks. The establishment of the model and optimal evaluation were verified by survival rate, pathological tests, biochemical tests, liver and spleen index calculation, immunohistochemistry, ELISA, and other assays. Results： The survival rate was 100% in the normal group, 46.15% in the low-dose group, 69.23% in the medium-dose group, and 84.61% in the high-dose group. The liver tissues of the rats in the normal group showed no abnormality to the naked eye; the liver of the rats in the low-dose group became darker in color, rougher in surface, with a small number of cancerous nodules and slightly hard texture; the liver of the rats in the medium-dose group was rough in surface, with several small cancerous nodules and scattered massive occupying nodules and hard texture; The liver of rats in the high-dose group became lighter in color, slightly rougher in surface, with no obvious cancerous nodules; HE staining showed that the liver tissues of rats in the low- and mid-dose groups were structurally disorganized, with large cellular heterogeneity and tumor cells. HE staining showed that the liver tissues of rats in the low and intermediate dose groups were structurally disorganized, with large cellular heterogeneity and tumor cell formation, while the structure of the liver lobules of the high dose group was unclear, with different degrees of edema, degeneration and necrosis of liver cells, and no obvious tumor cell formation was seen. Compared with the normal group, serum liver function (alanine aminotransferase, ALT), (aspartate aminotransferase, AST), (and total bilirubin, TBIL) were elevated in the low, medium, and high dose groups, and ALT and AST were significantly elevated in the low dose group, P＜0.05, the difference was statistically significant. The difference was statistically significant, ALT, AST and TBIL were significantly elevated in the middle dose group, P＜0.05, the difference was statistically significant, and the difference was not statistically significant in the high dose group, although it was elevated, P＞0.05; compared with the normal group, the serum (International Standardised Ratio, INR) level of the low dose group was significantly elevated, P < 0. 05, the difference was statistically significant; serum APTT (activated partial thromboplastin time, APTT), PT (prothrombin time, PT), AFP (alpha-fetoprotein, AFP ) were increased, P > 0.05, the difference was not statistically significant; serum APTT, PT, INR and AFP levels were significantly increased in the mid-dose group, P < 0. 05, the difference was statistically significant; serum PT and AFP levels were increased in the high-dose group, P < 0.05, the difference was statistically significant, and serum APTT levels were slightly increased, P > 0.05, the difference was not statistically significant; liver and spleen indexes were increased in the middle-dose group, P＜0. 05, the spleen index increased in the low dose group, P＜0.05, and the liver index increased in the high dose group, P＜0.05, the difference was statistically significant; the optical density value of liver tissue (alpha-fetoprotein, AFP) increased significantly in the low, medium and high dose groups, P＜0.05, the difference was statistically significant. Conclusions： Both the low and mid-dose groups could successfully induce the HCC rat model, but the pathological changes and biochemical findings of the mid-dose group were more in line with the pathogenesis of human HCC, and with fewer administrations and higher survival rate of the rats, it was possible to establish a more cost-effective and superior HCC model.
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Probing the anti-rheumatoid arthritis synovial neovascularization of alcoholic extracts of the Miao drug helleborus thibetanus franchon in rats based on the VEGF/VEGFR2 signaling pathway

aifei

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【Abstract】Objective This project aims to study the "Miao Medicine helleborus thibetanus franchon " as the research subject, investigating its anti-inflammatory activity in CIA rats and the regulatory mechanism on the VEGF/VEGFR2/P38MAPK pathway. Methods Sixty female Wistar rats were randomly divided into six groups: normal, model, positive control, and low, medium, high dose groups of helleborus thibetanus franchon , with ten rats in each group. Bovine type II collagen solution was injected into the tail of rats to replicate the rat model, and the positive drug group was given MTX2.0 mg/(kg-d) by gavage once every other day; the three dose groups of Iron Chopsticks, low, medium, and high, were gavaged with helleborus thibetanus franchon ethanol extract at 0.25 g/(kg-d), 0.5 g/(kg-d), and 1 g/(kg-d) once a day. The normal and model groups were given an equivalent volume of NaCl solution; continuous administration lasted for 28 days. During treatment, general conditions of the rats were observed, body weight changes recorded, and foot thickness measured. After treatment, the rats' hind limbs were taken for microCT to detect bone destruction, H&E staining for pathological study of the synovial membrane, immunohistochemistry to observe the number of neovascularization in the synovium, qRT-PCR to detect mRNA levels of VEGF-A, VEGFR2, TNFα in rat synovial tissue, and Western blot to detect expressions of VEGF, VEGFR2, p-p38, AKT, exploring the potential mechanism of Miao Medicine helleborus thibetanus franchon in treating rheumatoid arthritis. Results Compared to the normal group, the model group showed significant weight loss (P<0.01), increased foot swelling (P<0.01), visible proliferative synovial tissue with inflammatory cell infiltration, erosive lesions on bone surfaces, increased neovascularization in the synovium, and significant bone destruction in micro-CT, with reduced bone percentage, trabecular thickness, and bone density. Levels of VEGF-A, VEGFR2, TNFα mRNA, and proteins VEGF-A, VEGFR2, p-P38, p-AKT were significantly elevated (P<0.01). Compared to the model group, helleborus thibetanus franchon ethanol extract improved these conditions in a dose-dependent manner, with the high-dose group showing the best effect. There was a significant increase in rats' body weight (P<0.05), reduced foot swelling (P<0.05), ameliorated synovial and erosive bone lesions, reduced neovascularization in the synovium, and significantly lowered levels of VEGF-A, VEGFR2, TNFα mRNA, and proteins VEGF-A, VEGFR2, p-P38, p-AKT (P<0.01). Conclusion: Miao Medicine helleborus thibetanus franchon may alleviate joint inflammatory damage in CIA rats by modulating the VEGF/VEGFR2 signaling pathway, thereby exerting therapeutic effects on RA.
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Establishment of a mouse orthotopic lung cancer and lymph node metastasis model

HE Jie, ZHANG Xing

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【】 Objective In this study, a mouse orthotopic lung cancer and lymph node metastasis model was established to simulate the lymph node metastasis during the occurrence and development of lung cancer, and to study the immune cells in the microenvironment before lung cancer metastasis. Methods Mouse Lewis lung cancer cells were cultured, and mouse lung orthotopic transplantation tumor models were constructed by injection through chest wall cells. Samples were taken at 7, 14, 21 and 28 days after tumor transplantation, and the changes of orthotopic transplantation tumor, lymph node tissue structure and B cells were observed by HE staining and immunohistochemistry. Results The orthotopic lung cancer model of C57BL/6J mice was successfully constructed, and the mediastinal lymph nodes were obtained according to the anatomical location. The changes of germinal center, lymphatic vessels and blood vessels in the lymph nodes, the structural changes of lymph nodes with tumor metastasis and the infiltration of tumor cells were observed with the development of lung cancer through HE staining. IHC results also showed an increase in lymph node B cells (CD19+) density in the model group. Conclusions The establishment of this model lays a foundation for the future study of immune cells, especially B cells, in the microenvironment of lung cancer before lymph node metastasis.
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Oxidative damage and epithelial barrier disruption of lung tissue mediated by PM2.5 at different doses

shiyuyu, shaoqi, wangxueqian

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Objective To investigate the dose-dependence of inflammation, oxidative stress, and epithelial barrier disruption in C57BL/6 mice using different concentrations of PM2.5.Methods Thirty-six male C57BL/6 mice were randomly divided into control group, PM5.0 group, PM7.5 group, PM10.0 group, and treated with different concentrations of PM2.5 for 7 days.The control group was treated with intratracheal instillation of normal? saline.After the last exposure, the animals were sacrificed.The histopathological changes of lung tissues were observed using HE staining.The levels of interleukin-4 (IL-4), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in serum of the 4 groups of mice were detected using ELISA kits.The levels of NO, MDA, and SOD in serum and alveolar lavage fluid of the 4 groups of mice were detected using biochemical kits and ELISA kits.The apoptosis level of epithelial cells was observed using TUNEL staining.The expression of tight junction proteins in the epithelial barrier was detected using immunohistochemistry.Results Acute exposure to PM2.5 led to widened alveolar septa and inflammatory cell infiltration in the lungs of mice.The levels of inflammatory cytokines (IL-4, IL-1β, and TNF-α) in serum and NO, MDA, and SOD in lung tissues increased, while the expression of tight junction proteins in the epithelial barrier increased dose-dependently (P < 0.01 or P < 0.05). Conclusion Acute PM2.5 exposure causes acute lung injury in mice by activating lung tissue inflammation and oxidative stress, and PM2.5 exposure induces epithelial cell apoptosis and disrupts the tight junction of the epithelial barrier.This effect was most pronounced at a dose of 10 mg/kg.
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Aerobic exercise and empagliflozin alleviate ISO induced cardiac remodeling by inhibition of ferroptosis

QIN Di, QIN Xuelin, ZHENG Yiwei, LIN Yi, PENG Yong

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Objective The aim of this study was to explore the effect and possible mechanism of aerobic exercise and EMPA on ISO induced pathological cardiac remodeling. Methods The mice were randomly divided into Con group, ISO group, EX+ISO group, EMPA+ISO group and EX+EMPA+ISO group. The exercise group mice were continuously trained for 6 weeks, the EMPA group mice were continuously gavaged for 4 weeks, and all ISO group mice were subcutaneously injected with ISO for 7 days before dissection. The whole heart mass index, left heart mass index, heart mass to tibial length ratio, and left heart mass to tibial length ratio of mice were calculated through weighing and measurement. HE staining, Sirius red staining and WGA staining were used to observe the pathological changes, collagen fiber deposition and myocardial cell cross-sectional area in mice hearts. The qRT-PCR, Western blot and immunofluorescence staining were used to detect the expression of genes and proteins related to cardiac fibrosis and hypertrophy, macrophage infiltration, as well as the expression of genes and proteins related to ferroptosis and PI3K/AKT pathway in mice. Results (1) Compared with ISO group, EX+ISO group had a downward trend in whole heart mass index, left heart mass index, heart mass to tibial length ratio and left heart mass to tibial length ratio. EMPA+ISO group had a significant decrease in whole heart mass index and left heart mass index, heart mass to tibial length ratio and left heart mass to tibial length ratio were both down regulated. EX+EMPA+ISO group had a significant decrease in whole heart mass index, the other three indicators were all down regulated. (2) Compared with ISO group, the myocardial cells in three intervention groups were arranged more orderly, with a significant reduction in inflammatory cell infiltration, as well as a significant reduction in the area of cardiac fibrosis and the cross-sectional area of myocardial cells. (3) Compared with ISO group, the mRNA and protein expressions of Col 1 and Anp were significantly reduced in three intervention groups. The mRNA expression of Col 3 in EMPA+ISO group and EX+EMPA+ISO group was significantly reduced, and the mRNA expression of Col 3 in EX+ISO group showed a downward trend. (4) Compared with ISO group, the number of macrophages infiltration and the mRNA level of Il-6 were significantly reduced in three intervention groups. (5) Compared with ISO group, the mRNA levels of Nrf2 and Gpx4 were upregulated in three intervention groups, with a significant increase in GPX4 protein expression and a significant decrease in HO-1 protein expression. (6) Compared with ISO group, the mRNA level of Pi3k in EX+ISO group was significantly increased, the mRNA level of Pi3k in EMPA+ISO group and EX+EMPA+ISO group was upregulated. The mRNA level of Akt in three intervention groups showed an upward trend. The protein expressions of PI3K and p-AKT in EX+ISO group were significantly increased, and the protein expressions of PI3K and p-AKT in EMPA+ISO group and EX+EMPA+ISO group showed an increasing trend. Conclusions The moderate intensity aerobic exercise, the novel hypoglycemic drug EMPA, and their combination can alleviate ISO induced pathological cardiac remodeling, the mechanism may be related to the activation of the PI3K/AKT signaling pathway and inhibition of cardiac ferroptosis.
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Advancesinnon-surgicalanimalmodelsofvascular cognitiveimpairment

sunrong

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Vascularcognitiveimpairment(VCIvascularcognitiveimpairment)contains aseriesofillnessfrommildcognitiveimpairmenttodementia,comprehensive,attributableto cerebrovascular.Theestablishmentofappropriateanimalmodelsisverynecessaryforclinic opathologicalresearchanddrugdevelopment,butsofar,noanimalmodelcanperfectlysimulate thepathogenesisofVCI.Atpresent,carotidarteryandvertebralarterystenosisorocclusionare themainstreammethodsforVCImodeling,butinrecentyears,moreandmorenon-surgical methodshaveemerged,providingnewideasandprospectsforthestudyofthisdisease.Inthis paper,theconstructionmethod,modelmechanismandmodelcharacteristicsofVCInon-surgical animalmodelwerediscussed,soastoprovidereferenceforresearcherstochooseamoresuitable animalmodel.
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Progress in animal models of sick sinus syndrome

sunran, houping, xu guan zhen, bohuiying, sunyingying, zhangshuhan, wuyantong, liuyue

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Sick sinus syndrome (SSS) refers to the damage of the sinoatrial node and its surrounding tissues, which leads to the excitation and conduction dysfunction of the sinoatrial node, resulting in a series of arrhythmia diseases. It is of great value to better explore the pathogenesis of SSS, provide a basis for its treatment research, and establish an animal model that can simulate human sinus node dysfunction. In this paper, the selection of animals, the principles and methods of modeling, the evaluation methods and detection indicators of the model were reviewed, hoping to provide a basis for further study of the pathogenesis of SSS.
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Progress in the study of bone metastasis mechanism of prostate cancer based on a mouse model

zhangchenyang, huyaohua, zhangyanying, shichanghong

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【Abstract】 Prostate cancer is a prevalent malignant tumor within the male genitourinary system, characterized by a high propensity for bone metastasis. It is a leading cause of mortality, with approximately 70% of deaths attributed to this form of metastasis. Mouse models serve as a crucial tool in the investigation of prostate cancer""s bone metastasis, playing a pivotal role in elucidating the underlying pathophysiological mechanisms and in the development and assessment of therapeutic agents. In this review, we summarized the research progress of prostate cancer bone metastasis mouse models, construction methods, and evaluation strategies used in establishing mouse model. Particularly, this review emphasizes the exploration of the development mechanisms behind prostate cancer bone metastasis using mouse models. It aims to offer significant insights and serve as a valuable reference for f prostate cancer bone metastasis.
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Effects of acute lung injury on apoptosis of rat splenic T cells and the interventional effect of Yifei Jianpi Formula

YANG Yujie, ZHU Zhongbo, ZHANG Yanmei, LIU Xiping, ZHANG Xuhui, ZHANG Zhiming

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objective: To observe the apoptosis of splenic T cells and the expression of XAF1, FAS and TNF-α proteins in rats with acute lung injury (ALI), and to investigate whether the mechanism of protection against ALI by Yifei Jianpi Formula is related to the down-regulation of the expression of XAF1, FAS and TNF-α proteins, and the inhibition of T cell apoptosis.Methods: Sixty male SD rats of SPF grade were randomly divided into blank group, model group, positive group, and high, medium and low dose groups of Yifei Jianpi Formula. The positive group was given 0.5 g/kg dexamethasone by gavage, the high, medium and low dose groups of Yifei Jianpi Formula were given 12, 6 and 3 g/kg of Yifei Jianpi Formula by gavage, respectively, and the model group and the blank group were given equal amounts of saline by gavage, and the medication was administered once a day for 14d.The lung function of the rats in each group was detected by the animal lung function testing system, We observed the imaging characteristics of the lungs and changes in organ index and wet/dry weight (W/D) of lung tissues in each group of rats; hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung tissues in each group; flow cytometry was used to detect the splenic T-cell subpopulations (CD4₊^/CD_⁺8) and apoptosis of splenic T-cells in each group of rats and the Western blot technique was used to detect the levels of XAF1, FAS and TNF-α protein expression levels in the spleen.Results: The rats in the model group had reduced lung function, decreased spleen and thymus organ index, and significantly higher wet/dry weight (W/D) of lung tissue (P<0.01), Inflammatory exudation and alveolar rupture in the lung tissue, accompanied by thickening of the lung texture and large areas of ground-glass shadows, with a significant decrease in T-cell subsets (CD4₊^/CD_⁺8), and a significant increase in the expression of XAF1, FAS, and TNF-α proteins as well as in the rate of apoptosis of T-cells (P<0.01) After the intervention of the Yifei Jianpi Formula, the wet weight/dry weight (W/D) spleen of rat lung tissues was significantly reduced, the organ index of the thymus was significantly increased (P<0.05,P<0.01), the T-cell subpopulation (CD4₊^/CD_⁺8) was significantly increased, and the protein expression of XAF1, FAS, and TNF-α as well as the apoptosis rate of T-cells were significantly decreased (P<0.05,P<0.01)..Conclusion: ALI induced up-regulation of XAF1, FAS, and TNF-α protein expression and T-cell apoptosis in the spleen of rats, and Yifei Jianpi formula may protect against ALI by down-regulating XAF1, FAS, and TNF-α protein expression and inhibiting apoptosis of splenic T-cells.
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Treadmill exercise Up-Regulated BDNF/TrkB-CREB pathway to improve anxiety-like behavior in neuropathic pain rats

WANG Xiaoge, BAO Jinyu, YANG Shuai, LV Yihang, ZANG Weidong, LI Cui

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Objective This study aimed to investigate the effects of low-moderate intensity treadmill exercise on the pain and anxiety-like behaviors in rats with chronic constriction injury of the sciatic nerve (CCI) and to explore the neural mechanism of BDNF/TrkB-CREB pathway involved in exercise relieving pain and anxiety behaviors of CCI rats. Methods 48 SD rats were randomly divided into 4 groups: Sham group, CCI group, Sham+exercise (Sham+Exe) group, and CCI+exercise (CCI+Exe) group. The rats in the exercise group underwent treadmill training for 4 weeks. The paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were measured before and at different time points after the operation. The elevated plus maze (EPM) and open field test (OFT) were used to evaluate the anxiety-like behavior of the rats. The mRNA and protein expressions of BDNF, TrkB, and CREB in the hippocampus were detected by PCR and Western blot. Results (1) The PWT and PWL on the operative side of the rats in the CCI group were significantly lower than those in the Sham group at 7, 14, 21, 28, and 35 days after the operation (P < 0.01). Compared with the CCI group, the PWT on the operative side of the CCI+Exe group was significantly increased after 21 days (P < 0.05). The PWL on the operated side increased significantly after 14 days (P < 0.01). (2) The results of EPM showed that the percentage of time spent in the open arms in the CCI group was significantly lower than that in the Sham group (P < 0.01), and the percentage of time spent in the closed arms was significantly higher than that in the Sham group (P < 0.01). The percentage of open arm time in the CCI+Exe group was significantly higher than that in the CCI group (P < 0.05). (3) The OFT results showed that the percentage of time spent in the central area of the open field in the CCI group was significantly lower than that in the Sham group (P < 0.01), and the CCI+Exe group was significantly increased compared with the CCI group (P < 0.05). (4) Compared with the Sham group, the mRNA and protein expressions of BDNF, TrkB, and CREB in the hippocampus of the CCI group were significantly decreased (P <0.05, P < 0.01). Four-week treadmill exercise increased the mRNA and protein expressions of BDNF, TrkB, and CREB in the hippocampus of CCI rats (P < 0.05). Conclusions 4 weeks of aerobic exercise alleviates mechanical hyperalgesia and thermal hyperalgesia and alleviates the anxieties induced by chronic pain in CCI rats. Up-regulation of the BDNF/TrkB-CREB pathway may be one of the mechanisms of exercise in relieving chronic pain and improving anxiety.
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Comparing a rat model of pulmonary fibrosis brought on by bleomycin injections once or more intratracheally

Tang Huimeng, SONg Qinghua, XIE Yunyun, SUN Xin, YANG Fan, YANG Shuguang, TIAN Yange

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Objective A rat model of pulmonary fibrosis was constructed using single or two intratracheal drops of bleomycin (BLM) to compare the modeling rate and stability of the two modeling modalities. Methods A total of 150 SPF-grade SD rats were randomly divided into blank control group (Control group), single intratracheal drop of bleomycin (BLM-S group) and two intratracheal drops of bleomycin (BLM-M group). BLM (3 mg/kg, single time) was given by noninvasive intratracheal instillation in the BLM-S group. The rats in BLM-M group were intratracheal instilled with BLM (3 mg/kg, 2 mg/kg) on day 1 and day 14, and those in Control group were given intratracheal instillation of 0.9% sodium chloride injection (1 ml/kg). These rats were put to death in batches on the 28th, 42nd, 56th and 84th day after modelling. All of the rats were used to measure deep inspiratory capacity (IC), vital capacity (VC), static lung compliance (Cchord), and dynamic lung complication (Cdyn); The pathological changes of lung tissue were observed, and the extent of alveolitis and fibrosis was graded. Additionally, the expression of collagen-III (COL-III) in rat lung tissue was detected by immunohistochemistry. Results (1) General condition and survival: Respectively, The survival rates of Control, BLM-S and BLM-M groups were 100%, 80% and 66%. Rats in the BLM-S and BLM-M groups had substantially reduced body weights on days 14 to 42 compared to the Control group (P < 0.05, P < 0.01). Rats in the BLM-M group had substantially less body weight on days 28-42 than those in the Control and BLM-S groups (P < 0.05, P < 0.01). (2) Lung function: compared with the Control group, IC, VC, Cchord, and Cdyn were markedly decreased in the BLM-S group ( P < 0 .05, P < 0 .01) and IC, VC, and Cchord were significantly decreased in the BLM-M group ( P < 0 .05, P < 0 .01) on day 28; rats in the BLM-S group on day 42 had IC, VC, Cchord decreased significantly ( P < 0 .05, P < 0 .01); IC, VC, and Cchord decreased significantly ( P < 0 .05, P < 0 .01) in rats in the BLM-M group on days 42 to 84. (3) Lung pathology: inflammatory infiltration and fibrous cords appeared in BLM-S group from day 28 to 84, and then gradually decreased (P< 0.05, P< 0.01). In the BLM-M group, the fibrosis and alveolitis were relatively stable (P< 0.05, P< 0.01). (4) Collagen deposition: at all time points, the expression of COL-III in the lung tissue of rats in the BLM-S and BLM-M groups was significantly higher than that in the Control group (P < 0.05, P < 0.01); The content of COL-III in BLM-S group at 42-84 days was significantly lower than that at 28 days (P < 0.05). Conclusion Both methods are capable of effectively creating pulmonary fibrosis models. The single-dose approach is straightforward and has a reduced death rate, and the degree of fibrosis is clearly visible, in the 28 day, however, after 42 days, it progressively recovers. The two-time method instillation modeling has a greater success rate and better stability. Even, On the 84th day, over half of the rats still exhibited visible fibrosis.
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The effect of Cichorium glandulosum Boiss. et Huet on fecal bile acid profile in obese mice based on targeted metabolomics technique

qishuwen, zhongyewei, Adalaiti Abudurexiti, zhangrui, houwenhui, zhangchunzi, maxiaoyan, maxiaoli

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【】 Objective To investigate the effect of ethanolic extracts of chicory on the fecal bile acid profile of high-fat diet-induced obese mice using targeted metabolomics technology based on UPLC-Orbitrap-MS/MS. Methods Twenty-four 6-week-old C57 BL/6 male mice were selected and randomly divided into normal group, model group, drug administration group and metformin group. The normal group was given a regular diet, and the remaining three groups were fed daily with high-fat chow for 10 weeks to create an obesity model. After successful modeling, the dosed group was gavaged with 200 mg/kg ethanolic extract of Cichorium glandulosum Boiss. et Huet. daily and the metformin group was gavaged with 100 mg/kg metformin daily for 10 weeks.After 10 weeks, liver tissues were collected from the mice for the detection of hepatic total triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C); The bile acid metabolic profiles of mice in each group were further examined by ultra-high liquid chromatography-mass spectrometry (UPLC-MS/MS) in collected feces; Results Compared with the normal group, body weight (P<0.0001) and hepatic TG (P<0.05) of the model group mice were significantly elevated, while TC showed a trend of increasing (P>0.05), LDL-C showed a trend of decreasing (P>0.05), and HDL-C was significantly decreased (P<0.001),showing weight gain and abnormal lipid metabolism. The alcoholic extract of Cichorium glandulosum Boiss. et Huet.significantly reduced body weight (P<0.0001) and significantly decreased liver TG (P<0.0001) and LDLC (P<0.05) in mice. The results of methodological validation indicated that the UPLC-Orbitrap-MS/MS method established in this study could accurately quantify 52 bile acids in feces. Analysis of the concentration of each type of bile acids revealed that the alcoholic extract of Cichorium glandulosum Boiss. et Huet. significantly increased the secondary bile acid/primary bile acid ratio (P<0.05). The results of multivariate statistical analysis showed significant changes in bile acid metabolism pattern between normal, model, administered and metformin groups. Screening for differential bile acids in the administered group relative to the model group with VIP>1 and P<0.05 showed that the alcoholic extract of Cichorium glandulosum Boiss. et Huet.significantly elevated Lithocholic Acid in fecal bile acids (P<0.001, VIP=2.187), 23-Nordeoxycholic Acid (P<0.05, VIP=1.391), and significantly decreased Deoxycholic Acid in fecal bile acids (P<0.05, VIP=1.78). VIP=1.78), 3-Epideoxycholic acid (P<0.01, VIP= 1.881), Isolithocholic Acid(P<0.05, VIP=1.705),Cholic Acid-3-Sulfate (P<0.05, VIP=1.581), ω-Muricholic Acid(P<0.05, VIP=1.533), and Glycochenodeoxycholic Acid-3-O-β-glucuronide(P< 0.05, VIP=1.516), and by searching the KEGG database, it was found that differential bile acids were mostly involved in the secondary bile acid biosynthesis pathway. Correlation analysis showed that four differential bile acids, deoxycholic acid (rs=0.6445, P<0.001), Isolithocholic Acid (rs=0.5879, P<0.01), 3β-deoxycholic acid (rs=0.6649, P<0.001) and ω-Muricholic Acid (rs=0.5387, P<0.01), in feces were strongly and positively correlated with body weight. Conclusion Cichorium glandulosum Boiss. et Huet.alcohol extract exerts weight loss and ameliorates lipid abnormalities by regulating fecal bile acid metabolism.This may be due to the fact that Cichorium glandulosum Boiss. et Huet. alcohol extract regulates secondary bile acid biosynthesis and exerts a fat-reducing effect.
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Therapeutic effects of quercetin on the pain responses of a mouse model of paclitaxel-induced peripheral neuropathy and related mechanisms

Jin Ting, Li Peiyi, Nie Huimin, Yin Chengyu, Pan Yushuang, Zhu Zhihui, Liu Boyi, Liu Boyu

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Objective: To observe the effect of quercetin on mechanical allodynia, activation of astrocytes and upregulations of pain-related TRPV1 and P2X3 in mouse with paclitaxel-induced peripheral neuropathy. Method: 24 C57BL/6 mice were randomly divided into Control group, Model group, Model+Quercetin group, 8 mice per group. A total dosage 8 mg/kg of paclitaxel were injected intraperitoneally into mice of Model and Model+ Quercetin group to establish the model. Mice in Control group were injected intraperitoneally with the same volume of vehicle. At the 8th day after 1st injection, mice in Model+Quercetin group were injected with 60mg/kg quercetin solution orally and mice in other group were also injected with the same volume of vehicle. Mechanical pain were measured with von Frey. Immunofluorescence was used to detect the activation of astrocytes in the spinal dorsal horn and. Immunofluorescence and western blot were used to measure the expressions of TRPV1 and P2X3 in DRGs. Results: ① Compared with Model group, the mechanical pain of mice in Model+ Quercetin group were relieved. ②Compared with Model group, the activation of astrocytes and the expressions of TRPV1 and P2X3 in mice of Model+Quercetin group were alleviated (p<0.05). Conclusion: Our result indicate that quercetin can significantly reduce mechanical pain of paclitaxel-induced peripheral neuropathy. This mechanism maybe related to alleviating activation of astrocytes in spinal dorsal horn and reducing expressions of TRPV1 and P2X3 in the dorsal root ganglia.
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Safety and effect of Tibetan hemorrhoid ointment on rats with similar hemorrhoids

Zhuxinghao, Ren Qingjia, Zhuo Ga, Li Shanshan, Wu Zongyao, Miao Mingsan, Qi Dongmei

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Objective To investigate the acute toxicity, irritation and effects of Shexiang Anhe Hemorrhoids ointment on hemorrhoid-phased model rats via TLR4/p38 MAPK/NF-κB signaling pathway. Methods Sixteen New Zealand rabbits were randomly divided into the intact skin Shexiang Anhe Hemorrhoids ointment, the intact skin control group, the broken skin Shexiang Anhe Hemorrhoids ointment group and the broken skin control group, with four rabbits in each group. In the experimental group, 20 g of Shexiang Anhe Hemorrhoids ointment (1 g/mL) was evenly applied to the hair removal area of rabbits, and an equal volume of solvent (a mixture of glycerol, lanolin and water) was evenly applied to the hair removal area of the back of rabbits in the control group once a day for 14 d. Another 40 rats were taken and randomly divided into a normal group, a model group, a maillard group, and a hemorrhoid cream group of 10 animals in each group. The cream was applied once a day for 14 d. The acute toxicity of the cream in intact and broken skin of rabbits was observed by HE staining and other methods after treatment. In situ photographs were taken of the perianal tissues of rats with similar hemorrhoids to observe the efficacy of Shexiang Anhe Hemorrhoids ointment, and the length and width of the ulcers were measured with vernier calipers to calculate the area of the ulcers, and RT-qPCR was used to detect the expression of TLR4, p38 MAPK, and NF-κB mRNA in the perianal tissues of the rats. Results Compared with the control group with intact skin and broken skin, there was no significant change in the body mass of rabbits in the administered group; compared with the control group, the mean values of the irritation evaluation points of Shexiang Anhe Hemorrhoids ointment on rabbits with broken skin with the administration of the drug for 1 h, 24 h, 48 h, and 72 h were 1.5, 1, 0.5 and 0.25, respectively, and there was no obvious skin irritation.On the effect of hemorrhoidal similar rats, compared with the model group, Shexiang Anhe Hemorrhoids ointment group rats after 14 d administration of symptoms significantly reduced; rats ulcer area are significantly reduced, the difference is statistically significant (P<0.05); TLR4, P38MAPK, NF-κB mRNA levels were significantly reduced, the difference is statistically significant (P<0.05).Conclusions Shexiang Anhe Hemorrhoids ointment is a safe topical ointment with less acute toxicity and irritation to the skin, and has achieved good efficacy in the treatment of hemorrhoidal-adjacent rats, and its mechanism of action may be related to the inhibition of the TLR4/p38 MAPK/NF-κB signaling pathway.
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A comparative study of three ways to construct a rat model of cerebral hypoperfusion

Chen Qiu Yan

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【】 Objective: By comparing the cerebral blood flow, the morphological changes of nerve cells in brain tissue and the levels of serum oxidation and inflammatory factors in three different rat models of cerebral hypoperfusion, the experimental animal models were provided for the study of mechanisms and therapeutic drugs of cerebrovascular diseases and neurodegenerative diseases. Method: A total of 88 male SD rats were randomly divided into sham operation group (n=16), classic bilateral common carotid artery occlusion group (2-VO group, n=24), modified 2-VO group (n=24) and intraluminal thread technique group (n=24). Bilateral common carotid artery ligation was performed in the classic 2-VO group, while blood was drawn from the common carotid artery before ligation in the modified 2-VO group (1mL/100g). Middle cerebral artery occlusion was performed in the intraluminal thread technique group. In the sham operation group of the first two models, the common carotid artery was separated but not ligated, while the proximal end of the common carotid artery and the external carotid artery were ligated but not insert the bolt thread in the sham group of intraluminal thread technique model. Cerebral blood flow, infarct volume, serum inflammatory factor level, HE staining and ultrastructure of hippocampal tissue were measured at 1, 3 and 7 days after operation. Results: Laser speckle showed that the cerebral blood flow of rats of the three groups decreased rapidly after surgery, and then increased slowly. The decrease of cerebral blood flow of the modified 2-VO group was more obvious than that of the other two groups. At day 7, only the modified 2-VO group still had significant differences in cerebral blood flow compared with the sham group, and remained in a state of hypoperfusion (cerebral blood flow decreased by 30% compared with the operation). The results of TTC staining showed that the infarcts in the striatum of the three groups gradually increased with the extension of the time after operation, among which 4 rats (about 26.7%) in the modified 2-VO group had infarcts in both the cortex and striatum, while 10 rats (about 66.7%) in the intraluminal thread technique group. ELISA showed that the levels of inflammatory factors like TNF-α, IL-1β and hs-CRP in the three groups were increased after operation, and the levels of pro-oxidation factors ROS were also increased. In contrast, the level of antioxidant factor SOD decreased. At postoperative day 7, there was no significant difference in hs-CRP between the classic 2-VO and the intraluminal thread technique groups compared to the sham group. However, the modified 2-VO group still exhibited significant differences in all the above indicators compared to the sham group. HE staining showed that the modified 2-VO group caused more severe damage to the hippocampal CA1 and CA3 regions compared to the classic 2-VO and the intraluminal thread technique groups. Transmission electron microscopy demonstrated that the modified 2-VO group induced more severe damage to mitochondrial and endoplasmic reticulum in the hippocampal region compared to the the classic 2-VO and the intraluminal thread technique groups. Conclusion: Compared to the classic 2-VO and the intraluminal thread technique, the modified 2-VO method can induce more complete cerebral hypoperfusion and more severe neural damage within the same time frame, resembling the pathogenesis of human cerebral hypoperfusion more closely.
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A review of animal models of non-specific low back pain

ZHANG Qiang, DONG Bao-qiang, LIN Xing-xing, LIU Zi-wei, WANG Lei-Chao, ZHANG Feng, ZHANG Dan-ning, ZHANG Kai-xuan

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Nonspecific low back pain is a common clinical disease, whose pathogenesis and causes are still unclear, and the advantages and disadvantages of various therapeutic programs are controversial. Current research on this disease is mostly limited to clinical studies, and there is an urgent need to invest in a large number of animal experiments to analyze its underlying mechanisms. The construction of animal models is an important means to study the pathogenesis of non-specific low back pain and to explore therapeutic methods, but there are certain limitations and lags in the establishment of models for this disease. Therefore, this paper reviews the selection of animals, construction methods, and evaluation methods of relevant indexes of animal models of non-specific low back pain, and analyzes the advantages and disadvantages of various models, to clarify the existing problems of the current basic research of this disease, and tries to provide new ideas, aiming at laying a theoretical foundation for the study of the mechanism of non-specific low back pain and the improvement of the therapeutic strategy.
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Research progress of delirium animal models and evaluation methods

yangxiaotong, guolongfei, chenli, wangwenjuan, zhaoyinzhi, yuanyuan

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Delirium is an acute brain dysfunction syndrome characterized by confusion, confusion, and difficulty concentrating attention, which mainly affects ICU and elderly inpatients. Treatment is not only costly, but also leads to an increased risk of serious complications and death. Due to its complex etiology and unknown pathological mechanism, clinical drug treatment of delirium is largely ineffective. The construction of animal models is a powerful tool to understand the mechanism of delirium, screen new drugs and study intervention measures. This article reviews the experimental research related to delirium animal models at home and abroad in recent years, and summarizes the latest progress in the construction and evaluation methods of delirium animal models from three aspects: animal selection, model construction methods and model evaluation, so as to provide reference for experimental research based on delirium animal mode
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Establishment and evaluation of mice models of autoimmune thyroiditis with depression based on the NLRP3/Caspase-1/GSDMD pathway

LIZhengzheng, LIUZiyu, WANGZhimin, JINZhe, ZHAOZhuo, YANGXiao

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Objective: To prepare and evaluate an animal model of autoimmune thyroiditis with depression based on the NLRP3/Caspase-1/GSDMD pathway. Methods: 32 NOD.H-2H4 mices were randomly divided into normal group (N group), depression group (DP group), autoimmune thyroiditis with depression group (AIT+DP group), autoimmune thyroiditis group ( AIT group), 8 animals in each group. The N group was fed normally, the DP group was subjected to chronic unpredictable mild stress (CUMS) for 5 weeks, the AIT group was given 0.05% sodium iodide water to establish an autoimmune thyroiditis model, and the AIT+DP group was subjected to 5 weeks of chronic unpredictable mild stress (CUMS) on the basis of establishing the AIT animal model. 5 weeks of CUMS to establish the AIT+DP animal model. Evaluate whether the mouse autoimmune thyroiditis model has been successfully prepared by observing the thyroid tissue structure and lymphocyte infiltration of mice and serum TGAb and TPOAb levels; by measuring body weight, sugar water preference rate, open field behavior (central quadrant time, central quadrant time Proportion, number of standing, frequency of defecation, hair grooming time), and hippocampal pathological changes were used to evaluate the depression status of mice. If the model mice meet the above-mentioned indicators related to depression and autoimmune thyroiditis, it indicates that the AIT+DP animal model has been successfully prepared. Results: Compared with the N group, the serum TGAb and TPOAb levels in the AIT group and the AIT+DP group were significantly increased (P<0.01). A large number of inflammatory cells were infiltrated in the thyroid gland. The central quadrant time and central quadrant time of the mice in the DP group and the AIT+DP group Proportions, times of standing, frequency of defecation, and hair grooming time were reduced to varying degrees, glial cells in the cerebral cortex increased, neuronal cells decreased, accompanied by some nuclear atrophy, and the expression levels of NLRP3, IL-1β, Caspase-1, and GSDMD-N Significantly increased, especially in the AIT+DP group (P<0.01). Conclusion: 0.05% sodium iodide water and CUMS can better simulate the external performance and internal index changes of AIT+DP model animals, and can provide an animal model reference for research on AIT+DP disease.
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Progress in the study of animal models of dwarfism

Liu Yifan, Huang Nan, Liu Yuxuan, Yang Xi, Gan Lin

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Dwarfism is a globally rare growth disorder usually caused by genetics or disease, with the most prominent phenotype being short stature. Animal models are important tools for studying its pathogenesis, prevention, treatment options and identifying potential therapeutic targets and biomarkers. The development of genetic engineering technology has greatly promoted the application of gene-edited animal models in the study of dwarfism. In this review, we summarize and discuss the existing animal models of dwarfism from the theoretical basis, model characteristics and research applications for the reference of researchers and clinicians, so as to better carry out the research on the pathogenesis and prevention of dwarfism.
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Research progress on animal model of maternal separation and its related mechanism affecting learning and memory

LiNa, SHE Guifang, WANG Yaxin, LEI Qin, SUN Hongyan

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【】 Maternal separation is a kind of social deprivation in early life, which will damage the learning and memory ability of rats in adulthood. The animal model of mother-infant separation is one of the commonly used animal models to study the manifestations and mechanisms of learning and memory impairment. In this paper, the animal model of maternal-infant separation and the related mechanism of maternal-infant separation affecting offspring"s learning and memory are reviewed, in order to provide basis for subsequent related research on maternal-infant separation.
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Progress in resource utilization and model animal research of Daurian ground squirrel

XING Xin, FU Youtao, LIU Xinyu, YANG Ming

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Daurian ground squirrel (Spermophilus dauricus) is the main animal model used in hibernation researches. This species has many fantastic physiological adaptive characteristics, such as adaptation to extreme hypothermia, low metabolism, resistance to ischemia-reperfusion injury, muscle atrophy, and unique mechanisms of body fat accumulation. These characters make the Daurian ground squirrel as the unique model with important reference significance and research value in clinical and space flight applications. Application of Daurian ground squirrel as a model animal will help to further explore the physiological mechanisms mentioned above. In this review, we first introduced the biological characteristics and research values for clinical and space flight applications of the Daurian ground squirrels. Then, we introduced the current progress in the breeding and experimental animalization of the Daurian ground squirrels in the laboratory and field researches. Finally, we proposed precautions for the experimental animalization process of this species.
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Effects of Mosla chinensis seed oil the physiological behavior and antioxidant index of Drosophila were observed with Drosophila melanogaster as model animal

Xu Mengting, Zhu Yuchen, Su Dan, Song Yonggui, Zhang Wenkai, Xu Lei, Ma Qiuting, Liu Zhiyong

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Objective To investigate effects of Mosla chinensis seed oil on physiological behavior and antioxidant index in Drosophila melanogaster. Methods One-day-old wild-type Drosophila melanogaster was divided into control group, treatment group and model group. Drosophila model group was deprived of sleep by repetitive light stimulation at night to establish insomnia model, while the control group was exposed to the basal culture data. The negative geotaxis ability was used to explore the optimal concentration and period of drug treatment, and appetite test, negative geotaxis ability, stress test, olfactory memory test and detection of sleep-wake rhythm were used to explore the effects of Mosla chinensis seed oil on physiological behavior. In addition, activities of Super Oxidase Dimutase (SOD), Catalase (CAT) and Propanedial (MDA) were detected by enzyme-linked immunosorbent assay (Elisa). Results The Mosla chinensis seed oil enhanced the locomotion ability of 30-day-old Drosophila melanogaster (P < 0.01). Increased the activity of SOD and CAT (P < 0.01%) and decreased the content of MDA(P < 0.01); The sleep time of Drosophila was prolonged (P < 0.05 or P < 0.01%);Improved olfactory memory in fruit flies.After sleep deprivation, the night sleep time of female Drosophila model group decreased (P < 0.05%), and that of male drosophila model group decreased (P < 0.01). After feeding Mosla chinensis seed oil, the night sleep time of sleep deprived female drosophila was extended (P < 0.05), and that of male drosophila was extended (P < 0.01%). Conclusions Using Drosophila as model animal, it was found that Mosla chinensis seed oil had certain antioxidant ability in vivo, prolonging sleep time and improving olfactory memory of drosophila sleep-deprived drosophila.
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Changes in glycolipid metabolism, oxidative stress and myogenic differentiation level of skeletal muscle satellite cells in rats with type 2 diabetes mellitus intervened by exercise combined with probiotics

CHEN JuanJuan, ZHANG Yong Fa, MO WeiBin, LI GuoFeng, LI MingLiang

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OBJECTIVE: To observe the effects of exercise combined with probiotic intervention on glycolipid metabolism, oxidative stress and skeletal muscle satellite cell expression in type 2 diabetic rats.Methods: Sixty 8-week-old SPF-grade SD male rats, 10 rats were randomly selected as the normal control group (NC), and the rest of the rats were injected intraperitoneally with streptozotocin, of which 40 rats with blood glucose concentration ≥16.7 mmol /L were shown to have successful establishment of diabetes mellitus, and were randomly divided into the model group (TN), the exercise + diabetes group (YTN), the probiotic + diabetes group (GTN), and the exercise + After 6 weeks of aerobic treadmill exercise with incremental loads, glycemic and lipid metabolism indexes, oxidative stress indexes, and the expression of skeletal muscle satellite cell proteins were measured in rats.RESULTS: ①The levels of FBG, FINS, HBA1C and HOMA-IR in rats in the TN, YTN, GTN and YGTN groups were higher than those in the NC group (P < 0.01); the levels of FBG in rats in the YGTN group were lower than those in the TN, YTN, GTN and YGTN groups (P < 0.05); the levels of serum FINS in rats in the YTN, GTN and YGTN groups were decreased compared with those in the YTN, GTN and YGTN groups (P < 0.05); serum FINS levels of rats in YTN, GTN and YGTN groups were all decreased compared with those in TN group (P < 0.01); serum HBA1C levels of rats in YGTN group were all lower than those in TN, YTN and GTN groups (P < 0.01). The HOMA-IR indices of rats in the YTN and GTN groups were all lower than those in the TN group (P < 0.01), and the HOMA-IR indices of rats in the YGTN group decreased more significantly when compared with those in the TN, YTN and GTN groups (P < 0.01).②The serum T-CHO, TG, LDL and FFA levels of rats in the TN group were higher than those in the NC group (P < 0.05), and those of rats in the YGTN group were lower than those in the TN group (P < 0.05), and basically restored to the level of the NC group.The serum HDL levels of rats in the TN group were lower than those in the NC group (P < 0.05), and those of rats in the YGTN group The serum HDL level of rats in TN group was lower than that of NC group (P < 0.05), and that of rats in YGTN group was higher than that of TN group (P < 0.05), which was basically restored to the level of quiet group.③The serum MDA and 8-isoPGF2α levels of rats in the TN group were higher than those in the NC group (P < 0.05 or P < 0.01), and the serum MDA and 8-isoPGF2α levels of rats in the YGTN group were lower than those in the TN group (P < 0.05 or P < 0.01); the serum CAT, SOD, GSH-Px and T-AOC levels of rats in the TN group were lower than those in the NC group (P < 0.01), and serum CAT, SOD, GSH-Px and T-AOC levels of rats in the YGTN group were higher than those in the NC group (P < 0.01), but basically restored to the level at quiet time.④The protein expression of Pax7, MyoD, MyoG and Myf5 in the skeletal muscle of rats in the TN group was lower than that in the NC group (P < 0.05 or P < 0.01), and the protein expression of Pax7, MyoD, MyoG and Myf5 in the skeletal muscle of rats in the YGTN group was higher than that in the TN group (P < 0.05 or P < 0.01); basically, it was restored to the level of the NC group.In the TN group, the protein expression of MSTN protein expression was higher in the TN group than in the NC group (P < 0.05), and that in the YGTN group was lower than that in the TN group (P < 0.05); it was basically restored to the level of the NC group.CONCLUSION: It was shown that probiotics and aerobic exercise had a synergistic effect in improving blood glucose and insulin levels in type II diabetic rats, attenuating insulin resistance, improving lipids and oxidative radicals, and preventing the phenomenon of disturbed glucose metabolism in diabetic rats, thus achieving the regulation of the dynamic equilibrium within the organism. In addition, aerobic exercise combined with probiotics promoted myoblast differentiation, which played a role in preventing T2DM-induced loss of muscle mass and strength as well as muscle tissue complications.
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Liangxue-Jiedu decoction alleviates acute-on-chronic liver failure by inhibiting TLR4/JNK/NF-κB signaling pathway

LIU Zhengfang, ZHOU Wen, LI Zhenting, LI Qin, LIAO Naishun

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Objective To investigate the therapeutic effect of liangxue-jiedu decoction on acute-on-chronic liver failure (ACLF) model mice and its effect on Toll-like receptor 4 (TLR4) /c-Jun amino terminal kinase (JNK)/nuclear factor κB (NF-κB) signaling pathway. Methods An ACLF mouse model was established using the combined treatment of carbon tetrachloride (CCl₄), lipopolysaccharide (LPS), and D-galactosamine (D-GalN). Biochemical analysis was performed to evaluate liver function indicators including alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (TBIL). Histopathological examination was conducted to assess liver tissue morphological changes. Quantitative PCR was used to detect the mRNA expression of tumor necrosis factor α (TNFα), interleukin-6 (IL-6), interleukin-1β (IL1-β), and TLR4 in liver tissues. CCK8 assay was used to evaluate the interventional concentration of Liangxue-Jiedu decoction on Raw 264.7 cells. Enzyme-linked immunosorbent assay was used to detect the contents of TNF-α, IL-6 and IL1-β in the cell supernatant. Protein immunoblotting was performed to measure the expression of TLR4/JNK/NF-κB signaling pathway-related proteins, including TLR4, NF-κB, p-ERK1/2, ERK1/2, p-JNK and JNK. Results Compared with the ACLF model group, the Liangxue-Jiedu decoction reduced cell necrosis, fibrosis, and inflammatory cell infiltration in liver tissues, and decreased serum levels of ALT, AST and TBIL, as well as mRNA the expression of TNFα, IL-6, IL1-β, and TLR4. Liangxue-Jiedu decoction reduced TLR4/JNK/NF-κB signaling pathway-related protein expression in liver tissues. Furthermore, the in vitro result also showed that Liangxue-Jiedu decoction reduced TNFα, IL-6, IL1-β secretion in macrophage cells, and down-regulated the protein expression of TLR4/JNK/NF-κB signaling pathway. Conclusions Liangxue-Jiedu decoction effectively improved liver function in ACLF mice, which is closely related to the downregulation of the TLR4/JNK/NF-κB pathway.
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Study the effect of PM2.5 on hepatic lymphangiogenesis in C57BL/6J mice and metabolic-associated fatty liver disease model mice

DING Shibin, LI Yang, CHEN Yuping, JIANG Jinjin

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Objective To study the effect of PM2.5 exposure on hepatic lymphangiogenesis in C57BL/6J mice and metabolic-associated fatty liver disease model mice, and to provide a novel target for prevention and treatment of PM2.5-induced liver injury. Methods Forty male C57BL/6J mice were randomly divided into: control group, PM2.5 group, metabolic-associated fatty liver disease (MAFLD) group and PM2.5-MAFLD group. Mice in the MAFLD group and PM2.5-MAFLD group were fed with high-fat diet for 12 weeks, mice in the other groups were fed with normal chow diet. From 13 weeks to 16 weeks, mice in the PM2.5 group and PM2.5-MAFLD group were treated with PM2.5 by tracheal instillation (twice per week); and mice in the mice in the other groups were instilled with saline at the same time. All animals were killed 24 h after the last PM2.5 treatment. Serum ALT and AST were measured, and the expression of LYVE1 of liver tissues were?visualized using immunofluorescence staining. The levels of hepatic oxidative stress markers (4-HNE and GSH/GSSG) were measured. The protein expression levels of lymphangiogenesis markers (PROX1 and LYVE1), lymphangiogenesis regulatory protein VEGF-C and lymphatic junctional function VE-cadherin in liver tissue were determined using Western blot. Results PM2.5 exposure significantly increased the levels of serum AST and ALT, and obviously decreased the protein expressions of PROX1，LYVE1 and increased the protein expression of VEGF-C, and VE-cadherin in liver, as well as increased the level of 4-HNE and decreased the ratio of T-GSH/GSSG in liver of mice in the MAFLD group (P<0.05). However, PM2.5 exposure did not affect the levels of serum AST and ALT, the protein expressions of PROX1，LYVE1, VEGF-C and the level of 4-HNE and the ratio of T-GSH/GSSG in liver of the MAFLD model mice (P<0.05). Conclusions PM2.5 exposure obviously aggravated hepatic oxidative injury, reduced hepatic lymphangiogenesis through reducing VEGF-C in the liver of MAFLD model mice.
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Mechanism of Smilax glabra Roxb. flavonoids ameliorating ISO-induced cardiac hypertrophy and cardiac inflammation in mice

Fu danting

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【】Objective To study the effect and mechanism of Smilax glabra Roxb. flavonoids (SGF) on myocardial hypertrophy and cardiac inflammation induced by isoproterenol (ISO) in mice. Methods C57/6J mice were randomly divided into normal group, ISO model group (1.25mg/kg.d), SGF low-dose group (50mg/kg.d) and SGF high-dose group (100mg/kg.d). SGF administration groups were given prophylactic administration for 7 days before modeling, and then ISO subcutaneous injection was performed in each group, and echocardiography was performed after continuous injection for 7 days; Serum was separated from orbital blood, and the contents of NT-proBNP and inflammatory factors IL-1β, IL-6 and TNF-α in blood were detected. Myocardial specimens were collected and the pathological changes of myocardial tissue were observed, the myocardial ROS levels were detected by DHE staining. And the mRNA levels of heart-related hypertrophy genes and the changes in the expression of key factor proteins in the inflammatory grade pathway of NLRP3/Caspase-1/IL-18 were detected in myocardial tissue. Results SGF prophylactic administration decreased IVSd, IVSs, and EF in echocardiography and increased LVIDs, LVIDd, and ESV. The contents of IL-1β, IL-6, TNF-α and NT-proBNP in blood were decreased, and the mRNA expression levels of heart-related hypertrophy genes ANP, BNP and β-MHC were inhibited. The increase of myocardial ROS level was inhibited. The protein expression of NLRP3, Caspase-1(p20) and IL-18, which are key factors in the NLRP3/Caspase-1/IL-18 inflammatory pathway, was down-regulated in myocardial tissue. The hypertrophy and disordered arrangement of cardiomyocytes were improved, the increase of fibroblasts outside myocardial fibers was reduced, and the infiltration of inflammatory cells and fibrosis of myocardial tissue were alleviated. Conclusion SGF can improve ISO-induced myocardial hypertrophy and cardiac inflammation in mice, which may act through the NLRP3/Caspase-1/IL-18 inflammatory grade pathway.
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Experimental study on constructing the three-dimensional scaffold of rabbit cartilage shedding cells and evaluating their compatibility with stem cells

xujuju, xieyanyan, guozhiyi, mayukai, wanglinhong, yangmeirong, dengzhaoling, huabaolai, yanzhenyu

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Objective To prepare decellularized scaffolds from rabbit cartilage at various concentrations and assess their physicochemical properties and compatibility with stem cells, thereby providing an experimental basis for cartilage repair. Methods Bone marrow mesenchymal stem cells (BMSCs) were cultured using the Percoll density gradient separation method, followed by flow cytometric analysis and testing of their osteogenic and chondrogenic differentiation capabilities.Cartilage pieces were excised from rabbit knees and hip joints, subjected to physical crushing, repeated freeze-thaw cycles, and mixed enzymatic digestion for decellularization.To compare and observe the physicochemical properties of decellularized scaffolds at different concentrations, three groups were designed with concentrations of 100% (Group A), 50% (Group B), and 30% (Group C), with three replicates each.Third-generation PKH26-labeled BMSCs were seeded onto the optimally concentrated scaffolds and cultured for one week to observe cell growth. Results Flow cytometry detected BMSC surface antigens with positive expression of CD44 and CD90 and negative expression of CD45; osteogenic induction stained with alizarin red showed red calcific nodules; chondrogenic induction stained with alcian blue showed blue cartilaginous nodules; no apparent cell morphology was observed in the three groups of scaffolds stained with hematoxylin, eosin, and toluidine blue.There is a significant difference in DNA concentration between decellularized samples and non-decellularized scaffolds（P<0.05).The content of glycosaminoglycans is slightly lower than the normal values.Significant differences were observed between the three groups of scaffolds in terms of pore size, water absorption, porosity, tensile strength, and Young's modulus(P<0.05).After co-cultivation of stem cells with the scaffolds, cell adhesion was good. Conclusion Percoll density gradient separation can obtain high-purity rabbit BMSCs; the mixed decellularization method is more thorough. Group C (30%) scaffolds are the most suitable for constructing tissue-engineered cartilage repair.BMSCs cultured in vitro grew well on Group C scaffolds.
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Comparative study on SD rat models of rapid high-altitude entry under different hypoxia stress modes

Shen Dong-Shuai, Lu-Lu, Wang Hong-Yi, Zhang Mei, Chen Ke-ming, Niu Ting-Xian, Xiao-Pan

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Objective: To compare and study the acute high-altitude model established by SD rats under two types of hypoxic stress modes: high-altitude field and simulated high-altitude environment. Methods: SD rats were rapidly entered into a simulated high-altitude animal experimental chamber (4000 m) or a high-altitude field laboratory (4010 m) to establish a rat rapid high-altitude model. After 24 or 72 hours of exposure, physiological and pathological indicators related to high-altitude changes were collected and measured, mainly including blood routine, blood biochemistry, blood gas, oxidative damage indicators, inflammation indicators, and pathological tissue analysis. Finally, differential analysis was conducted on the results to obtain a differential evaluation report. Results：At the same altitude, both high-altitude field and simulated high-altitude exposure for 72 hours can cause significant lung and brain damage. Under the same exposure time, the blood routine, blood biochemistry, and blood gas results of animal bodies are similar. There was no significant difference in the detection results of inflammation indicators（IL-6, IL-1β, MCP-1 and IFN-γ）and oxidative damage indicators (MDA, SOD, and GSH). However, the PCO2 and BE of the simulated 72-hour group were significantly higher than those of other treatment groups, and the high-altitude field brain coefficient was significantly higher than that of the simulated high-altitude group. These results suggest that there may be slight differences between high-altitude field and simulated high-altitude environments. Conclusion: The simulated high-altitude animal experimental chamber can successfully establish a rapid high-altitude animal model. The simulated altitude can be appropriately increased on the basis of 4000 meters. If an altitude of 4000 meters is used, the exposure time should be greater than 24 hours but slightly shorter than 72 hours. If conditions permit, it is advisable to go to the plateau for on-site experiments as much as possible.
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Overview of animal models of non-steroidal anti-inflammatory drugs induced gastric ulcer

WANG Wen, HOU Yujun, SHI Yunzhou, WANG Lu, ZHENG Qianhua, ZHOU Siyuan, CHEN Ying, SUN Luqiang, CHEN Shuai, YAN Xiangyun, LI Yanqiu, LI Ying

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Gastric ulcer is a common digestive system disease, long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs) is the second most important cause of this disease. The NSAIDs-induced gastric ulcer animal models are the key to the experiments and are important for studying the pathogenesis, corresponding treatment methods, and effective mechanisms of NSAIDs-induced gastrointestinal injury. However, there is currently a lack of summary on the NSAIDs-induced gastric ulcer animal models. This review summarizes and compares the relevant literatures on animal researches on indomethacin and aspirin induced gastric ulcer in the past 10 years, including the selection of experimental animals, drug solvents, and specific modeling methods. It points out the limitations of current models, such as cumbersome modeling methods, incomplete modeling details, inadequate model fit for clinical use, and lack of comparative researches between drugs. Feasible solutions are proposed, aims to provide effective references for researches in this field.
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Application of Neurocircuit Identification Technology in Traditional Chinese Medicine Brain Science

Cui Yaru, Gao Qian, Li Zifa, Hu Minghui, Zhang Hao, Geng Xiwen, Wang Xinyu, Wei Sheng

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The neural circuit is the material carrier for the realization of brain function, consisting of a complex network of different neurons. Neural circuit identification technology tracks the structure and activity manipulation of specific neural circuits to study their adequacy and necessity for brain function, which is particularly crucial for understanding the pathogenesis of brain diseases. As a high-tech tool in the fields of neuroscience and brain science, it has been gradually introduced into the basic research of Traditional Chinese Medicine (TCM) in recent years. This article systematically reviews the principles of neural circuit identification technology and its application research progress in the field of TCM neuroscience. It is pointed out that the future development direction of this field should be based on the overall concept of TCM characteristics and the design of syndrome differentiation and treatment. Conducting research on the neural circuit mechanisms of diversified methods of TCM to intervene in diseases will help promote the deep integration of TCM and modern neuroscience.
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Study on the mouse model of aldosterone-induced multi-organ damage

LuoY, Zhang HT, Zheng YW, Meng XZ, Fang Z, Wang YT, FangZY

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Objective Establishment and evaluation of aldosterone-induced multiple-organ damage mouse model. Methods Twenty mice were randomly divided into four groups, with five mice per group: blank control Group A given 0 μg/(kg·d), low-dose aldosterone model Group B given 150 μg/(kg·d), medium-dose aldosterone model Group C given 300 μg/(kg·d), and high-dose aldosterone model Group D given 450 μg/(kg·d). An osmotic micro-pump containing aldosterone was surgically implanted under the skin that infused the mice with aldosterone for four weeks to establish an aldosterone damage model. The body weight and blood pressure of the mice were recorded weekly. After four weeks of modeling, blood pressure and histology in various organs were observed and analyzed. Results (1) After four weeks of infusion with aldosterone, the serum aldosterone levels of mice in groups C and D significantly increased, whereas there was no significant increase in Group B. (2) After the mice were implanted with osmotic pumps, the systolic blood pressure in each model group increased significantly in the second and third weeks, but in the fourth week, the blood pressure in the model groups decreased. (3) Renal and cardiac injuries, interstitial edema, collagen deposition, and fibrosis occurred to varying degrees in each model group. A small amount of collagen deposition occurred in the liver of Group B, and liver cell injury, collagen deposition, and fibrosis occurred to varying degrees in groups C and D. Conclusions Aldosterone can induce multi-organ damage in mice. Under this modeling method, organ damage mainly manifests as edema, collagen deposition, and fibrosis lesions.
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Research progress on experimental animal models of Huntington’s disease

FU Shuo, ZHANG Wen, SONG Junke, DU Guanhua

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Huntington’s disease (HD) is a type of autosomal dominant neurodegenerative disease, where the main symptoms in patients include chorea-like involuntary movements, psychiatric behavioral abnormalities, and cognitive impairments, severely affecting their lives and consuming extensive social and medical resources. To better understand the pathological mechanisms and explore treatment methods, a variety of HD experimental animal models have been successfully established. This article outlines the establishment and application of various animal models ranging from Caenorhabditis elegans, Drosophila melanogaster and zebrafish to mice, rats and miniature pigs, and analyzes the characteristics and advantages of different animal models. By reviewing different animal models and their relevant evaluation indicators, this article emphasizes the importance of utilizing a combination of multiple animal models to promote a deeper understanding of the disease mechanisms and develop effective treatment strategies.
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Analysis of Animal Models of Sequelae of Pelvic Inflammatory Disease Based on Data Mining

WANG Yuchao, ZHAGN Bing, PAN Jing, LI Yalin, YU Xiao, Liu Jinxing

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Objective:The characteristics of animal models of sequelae of pelvic inflammatory disease (SPID) were reviewed to provide a reference for standardizing the modeling process and improving the molding rate.Methods:The literature on animal models of SPID in the past 40 years was searched, and animal species, modeling methods, modeling cycles, modeling substances, positive control drugs, and evaluation indexes were summarized and analyzed.Results:A total of 243 literatures were included, most of which were induced by phenol paste method and microbial infection method to establish SPID model in rats. The modeling cycle was mainly 14-16 days, and the success of the model was mostly determined by pelvic tissue morphology observation and pathological HE staining. There are few researches on TCM disease combination model.Conclusion:The method of SPID animal model modeling lacks consistent criteria, so it is recommended to evaluate the changes of animal behavior, pelvic histomorphology and pathology. Secondly, TCM syndrome modeling lacks effective methods and evaluation standards, which need further research and development; Finally, the selection and use cycle of positive control drugs need to be further explored and perfected.
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Exploring the mechanism of action of sour jujube nut compound formula for depression based on network pharmacology and experimental validation

Zheng Hanwen, Liu Xinyue, Zhao Haiyan, Wang Jiayin, Luo Fulong, Fan Bei, Alberto Carlos Pires Dias, Wang Fengzhong, Wang Qiong

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Objective:This study was to use network pharmacology techniques to predict the key targets of Compound prescription of sour jujube kernel (ZSSF) for depression, and to verify its mechanism of action using a zebrafish model of rifampicin-induced depression. The drug targets of ZSSF were retrieved from the TCMSP database, and the target names were corrected using the UniProt database. Targets related to depression were identified using the GeneCards, OMIM, and NCBI databases. Protein-protein interaction information for the shared targets was predicted using the String database. The collected data was then analyzed using the Metascape database to determine the enrichment of GO and KEGG pathways. The results were visualized using microbiotics. Behavioral experiments and RT-qPCR experiments were conducted to verify the therapeutic effect of ZSSF on depression using a zebrafish depression model induced by Risperdal.Results: A total of 188 targets were screened to find the intersection of depression and ZSSF. The protein-protein interaction results showed that ZSSF primarily targeted TNF-α, IL-2, IL-6, IL-1β, and IL-10 for its antidepressant effect. KEGG pathway enrichment analysis revealed that ZSSF exerted its therapeutic effect on depression through various signalling pathways such as the TNF signalling pathway, PI3K-Akt signalling pathway, and cGMP-PKG signalling pathway. The results of the animal experiments showed that the treatment groups with high, medium, and low doses of ZSSF exhibited significant improvements in distance of movement under acoustic and light stimulation compared to the model group. The speed of movement was also significantly faster. Additionally, the mRNA expression levels of TNF-α, IL-2, IL-6, IL-1β, and IL-10 were up-regulated in the brain tissues of zebrafish in the high, medium, and low dosage groups of ZSSF compared to the model group.Conclusion: ZSSF exerts its antidepressant effect through multiple components and targets. Its antidepressant effect may be associated with the inhibition of inflammatory factors.
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Experimental study on the inhibitory effect of dendrobium on corneal neovascularization in rats after corneal alkali burn

Jiang Bo, Tian Sha, Li Tao, Li Chunxia, Zhou Xiaodong

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Objective To investigate the inhibition of dendrobium on corneal neovascularization (CNV) in rats after corneal alkali burn. Methods Animal models of corneal alkali burn were made in Sprague-Dawley(SD) rats, which were divided into normal control group, model control group, low-concentration gigantol group, high-concentration gigantol group and aflibercept group, with 10 rats in each group. The rats in low-concentration gigantol group, high-concentration gigantol group and aflibercept group were treated with 2.5 mg/0.05 mL gigantol, 5 mg/0.05 mL gigantol, 2 mg/0.05 mL aflibercept by subconjunctival injection after modeling. The CNV, corneal opacity score and the thickness of the cornea was observed and compared on the 3rd, 7th and 14th day after alkali burn. The ratio of CNV area to corneal area was calculated. On the 14th day, all rats were sacrificed. HE staining and immunohistochemistry were used to detect the expression of CD34 and VEGF. The proteins expression of VEGF, IL-1β and TNF-α was detected by ELISA. Results On the 3rd, 7th, and 14th day after alkali burn in rats, the percentage of CNV to total corneal area in low-concentration gigantol group, high-concentration gigantol group and aflibercept group were significantly smaller than that in model control group (all P < 0.05). On the 14th day, the corneal opacity score was lower in high-concentration gigantol group than that in model control group (P < 0.05). The corneal thickness in model control group and low-concentration gigantol group were significantly thicker than that in normal control group (both P < 0.001). However, the corneal thickness in high-concentration gigantol group and aflibercept group were not significantly different from that in the normal control group (both P > 0.05). In addition, compared with model control group, the protein expressions of VEGF, IL-1β, and TNF-α in corneal tissues in low-concentration gigantol group, high-concentration gigantol group and aflibercept group were significantly lower (all P<0.01). Conclusion Gigantol by subconjunctival injection can inhibit the formation of CNV in rats after alkali burn and promote the absorption of corneal edema.
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The effect of sex difference on the prevalence of inflammatory bowel disease in offspring and its mechanism

wuyuhong, chenbihua, wuhuan, liuhanwen, ouyangpuyu, mengxuanyi, chenhongbing

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Objective To investigate whether there is a sex difference in the offspring of Inflammatory Bowel Disease (IBD) mice with IBD. Methods BALB/c female mice were randomly divided into normal control and IBD groups. The mice in the normal control group drank autoclaved water freely, and the IBD group drank 2% Dextran sodium sulfate (DSS) for 7 days and then switched to autoclaved water for 10 days for 3-4 cycles, and at the same time, they were mated with healthy males. When the pups were 8 weeks old, they were divided into the Con group and IBD group, and the IBD group drank 3% DSS for 7 days. Disease activity index was scored by monitoring body weight, fecal consistency and blood in stool; pathological sections of colon tissues, changes in cup cells and mucus layer were observed; interleukin-6 (IL-6), IL-1β, IL-33, and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA); the real-time fluorescence quantitative PCR was used to determine the mRNA expression levels of tight junction proteins and MUC-2 in the colon. Results Compared with the IBD female mice, IBD male mice had higher DAI scores, significantly shorter colons, large inflammatory infiltrates, crypt abnormalities, and absence of cup cells in the colon, significantly lower relative mRNA expression of OCLN, significantly higher levels of IL-6 and IL-33, and significantly lower levels of IL-10. Conclusion The symptoms of colitis in the offspring of IBD mice were more severe in males than in females, which was mainly related to the severe impairment of the intestinal epithelial barrier function.
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Evaluation of Inhalation Anesthesia Induction for Tracheal?Intubation in Miniature Pigs

Leng Yan, Dai Na, Qiao qianqian, Zhao xiaoshuai, Tian Hao, Fan Mingxia

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Objective Medical Research cannot be separated from miniature pigs animal models. Due to the special anatomical structure of the oropharynx and the glottis is difficult to be exposured by laryngeal, the establishment of the airway is the most critical step in the perioperative period of miniature pigs. How to perform intubation while preserves spontaneous breathing has been one of the hot spots in the experimental research field of miniature pigs. This study intends to investigate the effect of small-dose Zoletil combined isoflurane inhalation in preserved spontaneous breathing endotracheal intubation of miniature pigs and to explore the minimum alveolar effective inhaled concentrations in 50% (MAC EI50) and minimum alveolar effective inhaled concentrations in 95% (MAC EI95) to provide a reference for clinical practice. Methods 26 miniature pigs were plan to be operated under general anesthesia. Anesthesia was conducted by Zoletil intramuscular?inject and isoflurane inhalation. Heart rate, blood pressure, respiration rate, body temperature, oxygen saturation, end-tidal CO2 partial pressure and end-tidal isoflurane concentration were monitored. The initial end-tidal isoflurane concentration was set 2.0% isoflurane increased or decreased according to a modified sequential method with an adjacent concentration ratio of 1.1. Tracheal intubation conditions and efficacy were evaluated by Cooper's score. MAC EI50, MAC EI95 and corresponding 95% CI of alveolar isoflurane concentration during endotracheal intubation with direct laryngoscope was calculated by Probit regression. Results The MAC EI50 of alveolar isoflurane concentration in miniature pigs during endotracheal intubation with direct laryngoscope was 3.07% (95% CI 2.70-3.61%), MAC EI95 was 3.82% (95% CI 3.41-7.49%). Conclusion Small-dose Zoletil combined with isoflurane inhalation is a better and safe anesthesia method for preserved spontaneous breathing endotracheal intubation in miniature pigs. The MAC EI50 of alveolar isoflurane concentration in miniature pigs during endotracheal intubation with direct laryngoscope was 3.07% (95% CI 2.70-3.61%), MAC EI95 was 3.82% (95% CI 3.41-7.49%).
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Gender difference in epileptic seizure and neuropsychiatric behavior abnormalities induced by kainic acid in rats

MA Mengdie, FAN Min, XU Songlin, ZHENG Qiang, HE Shuai, GE Jinfang

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Objective To observe the neuropsychiatric behavioral performance in kainic acid (KA)-induced epilepsy rats, based on which to investigate the gender difference in the acute seizure and behavioral performance in tasks of sense, motor, learning and memory in the remission phase, and explore the potential neurobiological mechanism. Methods Healthy SD rats aging 4 weeks were randomly divided into a control group and a model group, with 22 rats in each group, half male and half female. The epileptic rat model was induced by intraperitoneal injection of KA. The seizure latency and seizure frequency within 2 h of each rat were observed, seizure grade was assessed by Racine grade standard, and cortical electroencephalogram (EEG) was recorded. Behavioral performance were observed via a series of tasks including open field testing (OFT), balance beam walking, elevated plus maze (EPM), Y-maze, and novel object recognition (NOR). The level of GABA in the hippocampus was detected by ELISA, the injury of hippocampal neurons was observed by Nissl staining, and the protein expression of Synapsin-1 and Synaptotagmin1 in the hippocampus were detected by Western Blot. Result Both male and female rats presented typically epileptic behaviors after KA injection. However, compared with that of the male ones, the latency to the first seizure (P = 0.014) or grading Ⅳ-Ⅴ was advanced in female model rats (P < 0.01), and the frequency of epileptic seizures within 2 h was significantly reduced (P = 0.019). In the OFT, compared with the control group, KA induced epileptic model rats presented more motor but less hedonic behaviors, as indicated by the decreased moving distance in total and in the central area. Moreover, the grooming frequency was significantly reduced in the female model rats, as compared to not only the control, but also the male model rats (P < 0.01). The model rats spent more time to complete the task with higher score in the balance beam walking task, indicating a poorer ability of stability and balance. In the EPM, the exploration times in the closed arm of the male model rats was increased; The preference index of the novel arm or object was decreased in the Y maze and NOR, suggesting an impairment of learning and memory ability. Moreover, neuronal injuries were found in the hippocampus of the model rats, accompanied with a declined concentration of GABA and protein expression of Synapsin-1 and Synaptotagmin1, with no gender difference. Conclusion Intraperitoneal injection of KA could successfully induce an epilepsy rat model. However, there is a gender difference in not only the characters of acute seizures, but also the behavioral performance in sensory, motor, and learning memory during epileptic remission, Moreover, there was no gender difference about the hippocampal GABA concentration and expression of synaptic plasticity-related proteins, which could not declare clearly about the mechanism underlying the gender differences.
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Comparative study on the structure and function of aged macaca fascicularis hearts in different age groups

LIU Shu-hua, ZHOU Dan, LI Yun-feng, HUANG Zhong-qiang, GUAN Ya-lun, WU Chang-lin, LI Xue-jiao, LI Ge, ZHANG Yu

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Objective Conduct basic research on the structure and function of the heart in cynomolgus monkeys above 10 years old, in order to provide basic data for animal selection in elderly disease research. Methods A total of 144 cynomolgus monkeys above 10 years old were selected as the research subjects, including 37 females and 66 males in the 10-15 age group, and 21 females and 20 males in the 16-20 age group. The basic data of cardiac structure and function in middle-aged and elderly cynomolgus monkeys were obtained through comparative analysis of general indicators (body mass index, blood pressure, heart rate), blood biochemical indicators (blood glucose, blood lipids, and ion indicators), cardiac structure and function indicators. Results Comparing the general indicators between the 10-15 year old group and the 16-20 year old group, as age increases, the blood pressure and heart rate of female and male monkeys show an increase, while there is a significant difference in blood pressure changes between male monkeys. Comparison of two sets of blood biochemical indicators showed that as age increased, blood glucose, triglycerides, total cholesterol, low-density lipoprotein cholesterol, blood calcium, blood sodium, and blood potassium in female and male monkeys all showed an increase. Among them, blood glucose, triglycerides (male), total cholesterol, high-density lipoprotein cholesterol (male), low-density lipoprotein cholesterol (male), blood calcium, blood sodium, and blood potassium showed significant differences in changes. Lactate dehydrogenase showed a significant decrease. Comparison of cardiac contractile function between two groups showed that as age increased, the anterior and posterior diameters of the left atrium in both female and male monkeys significantly decreased. Female monkeys showed a significant decrease in interventricular septal end systolic diameter, left ventricular end diastole and systolic diameter, left ventricular end diastolic and systolic volume, and left ventricular mass index, while male monkeys showed no significant changes. Comparison of diastolic function between two groups showed that as age increased, the late diastolic velocity of the mitral valve in male monkeys decreased significantly. The early diastolic velocity of the left ventricular sidewall in female monkeys increased significantly. Correlation analysis was conducted between metabolic indicators and cardiac structure and function indicators of female and male monkeys, and the results showed that the correlation between metabolic indicators and cardiac structure and function indicators was weak in female monkeys, the maximum absolute Γ value does not exceed 0.39, and the correlation between metabolic indicators and cardiac structure and function indicators was relatively strong in male monkeys, the maximum absolute Γ value reaches 0.66. Conclusions Based on ultrasound diagnosis combined with metabolic indicators, the heart function of cynomolgus monkeys was studied, and basic data related to the structure and function of the heart in middle-aged and elderly cynomolgus monkeys were obtained. As age increases, blood glucose and lipid indicators in cynomolgus monkeys increase, while cardiac systolic and diastolic functions show a downward trend, similar to changes in the middle-aged and elderly population. This provides data support for animal selection for research on age-related diseases related to heart function.
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Establishment of a Grading Model for Acute Gastric Mucosal Injury in Rats based on Cutaneous Neurogenic Exudation Response

Dong Lin, Shi-yi Qi, Jin-wen Lin, Shi-hao Wang, Li-li Lin

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Objective: This study aims to establish an acute gastric mucosal injury (AGMI) rat model induced by different concentrations of hydrochloric acid (HCl) and investigate the effects of intravenous injection of different concentrations and doses of Evans Blue (EB) on the survival rate and surface exudation of the model. Methods: Study 1: Wistar rats were randomly divided into five groups based on body weight: 150-180g group, 180-200g group, 200-250g group, 300-400g group, and 400-500g group, with a total of 40 rats in 8 subgroups based on HCl concentration (0.40M, 0.45M, 0.50M, 0.55M, 0.60M, 0.65M, 0.70M) and a control group treated with physiological saline, with 3 rats in each group and a total of 120 rats. The survival rate of rats in each group was evaluated after modeling for 24 hours, and the interaction between body weight and HCl concentration on rat survival was analyzed. On the basis of determining the five gradient HCl concentrations, the pathological changes of gastric mucosa were observed under microscopy using hematoxylin-eosin staining. Study 2: The highest concentration of HCl was selected for model preparation, and AGMI rats were randomly divided into six groups based on different concentrations and doses of EB: 0.5% (0.4ml) group, 1% (0.1ml/100g) group, 1% (0.2ml/100g) group, 2% (0.1ml/100g) group, 2% (0.2ml/100g) group, and 5% (0.1ml) group, with 5 rats in each group and a total of 30 rats. The survival rate (24 hours after injection) and degree of surface exudation of rats in each group were evaluated. Results: Study 1: The symptoms of AGMI rats after modeling were positively correlated with HCl concentration, and the 24-hour survival rates of rats in the 0.65M and 0.70M groups were both 0% across all body weight groups. The Kaplan-Meier survival curves of rats of different body weights showed significant differences in survival rates among rats treated with different concentrations of HCl (P<0.00). The interaction analysis showed a significant interaction between HCl concentration and body weight in terms of observed survival time (P<0.00). The histopathological observation of gastric mucosa in rats treated with the determined five gradient HCl concentrations (0.40M, 0.45M, 0.40M, 0.45M, 0.50M, 0.55M, and 0.60M) showed that the gastric mucosal tissue of rats in the physiological saline group had intact structure without inflammatory cell infiltration. The degree of inflammatory cell infiltration in AGMI rats was positively correlated with HCl concentration. Study 2: In AGMI rats prepared with 0.60M HCl, the survival rate after intravenous injection of 5% EB (0.1ml) was only 40% at 24 hours. The Kaplan-Meier survival curves showed no statistically significant difference in survival rates among AGMI rats treated with different concentrations and doses of Evans Blue (EB) (P>0.05). The results of surface exudation in AGMI rats after intravenous injection of EB showed that rats in the 0.5% (0.4ml) group did not have any color change in the skin and eyes, and had fewer surface exudation points, or even no exudation points. Rats in the other groups had more obvious exudation points. Among them, rats in the 2% (0.1ml/100g) and 2% (0.2ml/100g) groups had better skin color and exudation conditions. The skin color of rats in the 2% (0.2ml/100g) group further deepened over time at 24 hours after EB injection (28 hours after modeling). The results of surface exudation in AGMI rats after intravenous injection of EB showed that rats in the 0.5% (0.4ml) group did not have any color change in the skin and eyes, and had fewer surface exudation points, or even no exudation points. Rats in the other groups had more obvious exudation points. Among them, rats in the 2% group had better skin color and exudation conditions. The surface exudation at 24 hours after EB injection further showed that the skin color of rats in the 2% (0.2ml/100g) group deepened over time. Conclusion: Modeling with HCl can achieve the establishment of multiple precise concentration gradients. Fasting rats with a body weight of 180-200g can be administered HCl at concentrations of 0.40M, 0.45M, 0.50M, 0.55M, and 0.60M to prepare the AGMI model. Intravenous injection of 2% EB (0.2ml/100g) will be conducive to the study and analysis of the distribution characteristics of surface exudation points in AGMI rats over time and with changes in the condition.
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Study on the preventive and therapeutic effect and mechanism of Caryopteris incana on primary dysmenorrhea rats with cold coagulation and blood stasis syndrome

chenmeian, huangfeng, liangxiaoqing, lanxiaoqi, cuixiangxiang, zengxueqi

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Objective To explore the prevention and treatment effect of Caryopteris incana decoction on primary dysmenorrhea rats with cold coagulation and blood stasis syndrome. Methods Sixty healthy SPF-grade SD female rats were randomly divided into normal group, model group, ibuprofen group, high-dose group of Caryopteris incana, medium-dose group of Caryopteris incana and low-dose group of Caryopteris incana, with 10 rats in each group. Except for the normal group, the other groups were treated with cold stimulation combined with estradiol benzoate and oxytocin to establish a rat model of primary dysmenorrhea with cold coagulation and blood stasis syndrome. On the 5 th day of modeling, the rats were given intragastric administration for 10 days. The symptoms of rats were observed and recorded. The writhing response of rats was measured. The hemorheological indexes of rats were measured. The levels of plasma thromboxane B2 (TXB2), 6-keto-prostaglandin F1α (6-Keto-PGF1α), estradiol (E2) and progesterone (PROG) in serum of rats were measured. The levels of prostaglandin F2α (PGF2α), prostaglandin E2 (PGE2), NO and Ca2+^{in uterine}tissues of rats were measured. The organ indexes of uterus and ovary were calculated and the histopathological changes were observed. Results Compared with the model group, the symptoms of the rats in the high and middle dose groups were significantly improved. Each dose group of Caryopteris incana decoction could significantly prolong the latency time of writhing reaction (P＜0.01) and reduce the number of writhing times (P＜0.01). The high and medium dose groups of Caryopteris incana decoction could significantly reduce plasma viscosity, hematocrit of rats, and the whole blood viscosity at high, medium and low shear rates (P＜0.01). The high and medium dose groups of Caryopteris incana decoction could significantly reduce the levels of TXB2 and E2 and E2/PROG ratio in serum of rats and increase the levels of 6-Keto-PGF1α and PROG in serum of rats. The high and medium dose groups of Caryopteris incana decoction could significantly reduce the uterine index and ovarian index of rats (P＜0.01, P＜0.05). The high and medium dose groups of Caryopteris incana decoction significantly improved the morphological lesions of uterus and ovary. The high and medium dose groups of Caryopteris incana decoction could significantly reduce the levels of PGF2α, Ca2+ ^andPGF2α/PGE2 ratio in uterine tissue of rats (P＜0.01, P＜0.05) and and increase the levels of PGE2 and NO in uterine tissue of rats (P＜0.01). Conclusion Caryopteris incana decoction can effectively improve the clinical symptoms of primary dysmenorrhea rats with cold coagulation and blood stasis syndrome, and has a good control effect. Its mechanism may be correlated with the levels of TXB2, 6-Keto-PGF1α, E2 and PROG in serum and PGF2α, PGE2, NO and Ca2+^{in uterine tissue.}
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Effect of acute ozone exposure on miRNA expression profile of exosomes in alveolar lavage fluid of rats

yangqingcheng, zhaojiao, yangrenxin, yanzhangong, xizhuge, liuxiaohua

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Objective To clarify the changes of microRNA ( miRNA ) expression profile in rat alveolar lavage fluid-derived exosomes after acute ozone exposure, and to analyze its potential relationship with lung injury. Methods Adult male Wistar rats were exposed to 0 ppm, 0.5 ppm and 2 ppm ozone for 6 h in a gas exposure chamber. After 24 h, the rats were anesthetized, alveolar lavage fluid was collected and exosomes were extracted. The differentially expressed miRNAs in exosomes were detected by miRNA-seq technology. The miRDB databases were used to predict the target genes of differentially expressed Top 10 miRNAs between groups, and GO analysis and KEGG analysis were performed on the target genes. The differentially expressed miRNAs with high expression abundance and good uniformity in the group were verified by qRT-PCR. Results Compared with the 0 ppm group, there were 8 up-regulated and 55 down-regulated miRNAs in the 0.5 ppm group, while 32 up-regulated and 61 down-regulated miRNAs in the 2 ppm group. Compared with the 0.5 ppm group, 90 exosomal miRNAs were up-regulated and 74 were down-regulated in the 2 ppm group. The results of enrichment analysis showed that the target genes of differentially expressed Top 10 miRNAs were mainly involved in regulating DNA binding, heterocyclic compound binding and affecting transcription, and were closely related to cancer, MAPK, PIK3-AKT, TNF, ErbB, mitophagy and FoxO signaling pathways. The results of qRT-PCR showed that the sequencing results were basically reliable. Conclusion Acute ozone exposure can lead to changes in the expression profile of exosomal miRNA in alveolar lavage fluid of rats. Differential miRNA may be involved in pathological processes such as lung tissue inflammation and lung injury caused by ozone exposure.
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Overview of experimental methods for behavioral studies of Parkinson's disease in rats and mouse and commonly used animal models of Parkinson's disease

huyilong, zhaoyinan, miaojinxin, miaomingsan

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Parkinson's disease is the second most common neurodegenerative disease in middle-aged and elderly people, with a long disease cycle and complex treatment process, which is a great challenge in today's society. Behavioral changes in animal models of Parkinson's disease can intuitively reflect the modeling situation of experimental animals and the changes after drug intervention. Therefore, the selection of standardized animal models and appropriate behavioral assays is the basis for Parkinson's disease mechanism research and anti-Parkinson's disease drug development. In this paper, we summarize the Parkinson's behavioral experimental methods in mice and rats at home and abroad, and systematically summarize the experimental equipment, experimental methods, evaluation indexes, and precautions of commonly used Parkinson's behavioral experiments, as well as provide an overview of the commonly used Parkinson's models, and analyze the modeling mechanism, the match with the clinical features of Parkinson's disease, and the advantages and disadvantages, which can help researchers choose the appropriate Parkinson's model according to the research purpose. It can help researchers to choose appropriate Parkinson's models and behavioral testing methods according to the purpose of the study, which is of some significance to the in-depth study of Parkinson's.
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Advances in the study of the role of glycyrrhetinic acid and its derivatives in neurodegenerative diseases

LIU Yixiao, HAN Xinyuan, XIAO Yitong, YU Xinzhuo, YE Tianyuan

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Accompanied by the aggravation of global population aging, the incidence of neurodegenerative diseases has been increasing year by year, which seriously affects the quality of life of elderly patients and imposes a heavy burden on the society. Glycyrrhetinic acid is one of the main active ingredients of the traditional Chinese medicine Glycyrrhiza glabra, which inhibits neuroinflammation and protects neurons, etc. There is an increasing number of studies on the mechanism of action of glycyrrhetinic acid and its derivatives in neurodegenerative diseases. In this review, the studies on the effects and mechanisms of action of glycyrrhetinic acid and its derivatives in Alzheimer"s disease, Parkinson"s disease, amyotrophic lateral sclerosis, multiple sclerosis, and cerebellar atrophy are summarized, and on the basis of which, the future applications of glycyrrhetinic acid and its derivatives in neurodegenerative disorders are discussed and prospected.
Keyword:glycyrrhetinic acid;derivative;mechanism of action;neurodegenerative disease;progress of research
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Effect of galectin-3 gene knockout on abscess development and mast cell activation in MRSA-infected mouse skin model

Wang shu jun, Zhang ding, Li yi ming, Zhang si yi, Zhou jing, Chen zi han, Cheng mei qi, Han shan shan, Wang de cheng, Chao jin

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【Abstract】Objective The study aims to investigate the effects of Galectin-3 (Gal3) on the skin abscess development and activation of mast cell (MC) in mice skin infected with methicillin-resistant Staphylococcus aureus (MRSA). Methods Gal3 knockout (Gal3-/-) mice, at 6-8 weeks of age, were subcutaneously injected with MRSA or the same volume of phosphate buffer saline (PBS) PBS, five mice per group. The development and pathological changes of skin abscess were monitored and recorded. The bacterial load in skin tissues with bacterial culturing, the expression of associated cytokines, morphological differences of MC, and the distribution of MC activation marker 5-hydroxytryptamine (5-HT) were detected respectively. Results the inoculation area of the skins in both WT + MRSA and Gal3-/- + MRSA groups showed progressive abscess, accompanied by abscess exudation after subcutaneous infection with MRSA. However, compared to WT + MRSA group, Gal3-/- + MRSA mice got smaller abscess areas (P<0.01) and less infiltrating inflammatory cells in the skin with histopathological observation. The bacterial load in the skin tissues of Gal3-/- + MRSA group was significantly lower than that of WT + MRSA group (P<0.01). Meanwhile, the cytokines including IL-1β, TNF-α, IL-33, TGF-β and IL-10 were significantly less (P<0.05) in Gal3-/- + MRSA mice than that in WT + MRSA mice (P<0.05). MC were active and degranulated in the skin tissue of the WT + MRSA mice, while Gal3-/- + MRSA mice got less degranulated mast cells than WT + MRSA mice (P<0.05). Immunohistochemical staining showed that the expression of 5-HT was significantly increased in the WT + MRSA group, while decreased when Gal3 was deficient in mice. Conclusion Gal3 deficiency can reduce the activation and degranulation of mice skin MC after MRSA infection, which changes inflammatory responses and alleviates the abscess of skin tissue.
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The use of ferrets in non-clinical studies of anti-infective drugs

liuxuewu, tangzining, pengdongdong, duyanhua, jiangdejian

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The advantage of ferrets in non-clinical studies of anti-infective drugs is that pathogenic microorganisms (especially viral strains) can infect and spread without host adaptation, and the clinical symptoms of infected animals are very similar to those of humans. Although ferrets have played a very important role in the research and development of antiviral drugs, their application scope is still limited, which may be related to the lack of corresponding national standards for laboratory animal feeding and application of ferrets, and the lack of specific diagnostic and detection reagents. This paper summarized the characteristics of ferrets as infectious disease models, and summarized and analyzed the application direction of ferrets in anti-infective drug research, aiming to promote the further standardization and standardization of the use of ferrets.
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Isorhamnetin inhibit the inflammatory apoptosis of AKI tubular cells by inhibiting LncRNA33782 to block the NF-κB/Bcl-2/Bax signaling pathway

Jia Jian, Tan Ruizhi, Zhong Xia, Su Hongwei, Wang Li

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Objective To investigate the role of LncRNA-33782 in AKI tubular injury and the mechanism by which Isorhamnetin ameliorates AKI tubular inflammatory cell apoptosis. Methods Twenty-four male C57BL/6J mice were randomly assigned into four groups: control group, AKI model group, electroporation + AKI group, and treatment group (30 mg/kg orally administered Isorhamnetin). Renal function, histopathological alterations, and renal inflammatory cytokines (IL-1β, IL-6, TNF-α) were evaluated to assess the role of LncRNA-33782 in AKI and the therapeutic effect of Isorhamnetin following electroporation-mediated LncRNA-33782 knockdown and Isorhamnetin treatment in mice. Subsequently, an in vitro experiment involving LncRNA-33782 overexpression was conducted to investigate the mechanism of Isorhamnetin treatment. NF-κB was considered a key signaling pathway mediating inflammation, while Bax, Bcl-2 protein expression levels, and flow cytometry apoptosis detection results were employed to assess tubular cell apoptosis. Results Renal injury and abnormal renal function significantly improved after electroporation-mediated LncRNA-33782 knockdown, accompanied by a marked decrease in inflammatory cytokine expression levels. Isorhamnetin treatment also notably attenuated AKI-induced renal injury, downregulated LncRNA-33782 expression, inhibited the NF-κB signaling pathway, and alleviated tubular cell apoptosis. However, Isorhamnetin"s therapeutic effect on AKI was suppressed after in vitro LncRNA-33782 overexpression. Conclusions LncRNA-33782 promotes AKI renal inflammatory response, while Isorhamnetin can inhibit AKI tubular inflammatory cell apoptosis mediated by LncRNA-33782-induced NF-κB activation.
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Comparison of guinea pig tuberculosis models established by two respiratory infection routes

LI Xinyu, LI Haifeng, WANG Yu, QU Peijie, WANG Junfei, ZHAN Lingjun

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Objective: This study aims to explore and compare guinea pig tuberculosis models using different doses of intranasal and aerosol infection routes, providing a foundation for establishing a standardized guinea pig tuberculosis model of respiratory tract infection. Methods: 24 female guinea pigs were randomly divided into 6 groups, with 4 guinea pigs in each group. The guinea pigs were infected with Mtb through either aerosol or intranasal at two doses respectively, and were equally divided into 3 groups (A/B/C, D/E/F). The general manifestations of the guinea pigs were observed after infection. All guinea pigs were dissected on the 14th day, and lung, spleen, and liver tissues were obtained for gross examination as well as histopathological analysis using HE staining to identify characteristic lesions associated with tuberculosis. Acid-fast staining was performed on in situ tissues and organs followed by bacterial culture to analyze bacterial load. Results: The guinea pigs in the four infection groups (B-C, E-F) exhibited macroscopic tuberculosis lesions in the lung, spleen, and liver. Histopathological examination revealed the presence of tuberculous granuloma lesions. Acid-fast staining and bacterial load analysis demonstrated that the bacteria were primarily localized in the lung tissue of the aerosol-infected group (B/C), with a few also present in the spleen and liver, and the bacterial load ranged is 10^4-5 CFU/ml. In the intranasal infection group (D/E), bacteria were found in the lung, spleen, and liver with a similar bacterial load of 10^4-5 CFU/ml. There was no significant difference observed in lesion severity or bacterial load between groups B-C and E-F; however, groups B, C, and F displayed low standard deviations for both pathology and etiology. Conclusion: A guinea pig model of acute tuberculosis has been successfully established using two different doses administered through distinct routes of infection. Pathological examination and pathogenic analysis demonstrated that an aerosol dose of 500 CFU effectively prepared a homogeneous model of acute tuberculosis with good consistency among subjects. Additionally, intranasal infection with 50,000 CFU Mtb also produced a relatively uniform model of tuberculosis; however, it should be noted that an aerosol infection at 500 CFU developed into an acute tuberculosis model more rapidly compared to intranasal infection at 50,000 CFU.
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The establishment and utilization of a fluorescent living genetically engineered mouse model of glioma-mural cells using a two-photon microscope

MA Chengyan, YANG Xingjiu, SHI Xudong, GAO Ran

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Objective The aim of this study was to develop and assess a genetically engineered mouse model for visualizing in vivo fluorescence of glioma, mural cells, and blood vessels using two-photon microscopy. Methods PDGFRβ-Cre(+/-):Rosa26-tdTomato(+/-) genetically engineered mice were underwent skull clearance and injected with GL261-CFP. This was done to study the dynamic alterations in blood vessels and mural cells during the progression and invasion of glioma using a two-photon microscope. Results PDGFRβ-Cre(+/-):Rosa26-tdTomato(+/-) mice were successfully bred and subjected to HE section analysis of functional organ tissues, revealing no discernible differences from C57BL/6 mice in terms of appearance and morphology. Cre recombinase activity was fully induced following Tamoxifen treatment on day 7. Subsequent GL261-CFP inoculation demonstrated the dynamic progression of glioma proliferation and invasion, as well as vascular abnormalities and increased mural cell detachment within the tumor. Conclusions The genetically engineered mice that mural cell expressing fluorescent were bred successfully. Utilizing fluorescein isothiocyanate-dextran labeling of blood vessels and blue fluorescent tumor cells, glass discs and fixed rings were employed to replace the skull of the mice. This allowed for the tracking of morphological and structural changes in blood vessels and vascular supporting cells following the development of brain tumors in vivo over an extended period. This model offers a valuable tool for studying brain diseases through pathological visualization.
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Establishment of Ifitm3 knockout mice and exploring the effects of Ifitm3 on proliferation and differentiation of mouse neural stem cells

wang kaiyu, lei xuepei, huang yiying, shi guiying, yue hanwei, wang jie, lin yifan, tang jiaming, bai lin

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Objective To establish interferon-induced transmembrane protein 3 (Ifitm3) knockout mice for exploring the effects of Ifitm3 on the proliferation and differentiation of mouse neural stem cells. Methods The Ifitm3 knockout mice were established by CRISPR/Cas9 method and identified by PCR and Western Blot. The phenotypic differences between the Ifitm3 knockout mice and wild-type mice were analyzed by immunohistochemistry and flow cytometry. Adult mouse neural stem cells were isolated and cultured, the number and size of neurospheres were detected, and markers of proliferation and differentiation of neural stem cells were detected by RT-qPCR, Western Blot and immunofluorescence. Results Ifitm3 knockout mice were successfully established, and the Ifitm3 knockout mice developed normally, and there were no obvious abnormalities in histopathology and immune system. In vitro experiments showed that Ifitm3 affected the self-renewal potential of mouse neural stem cells, led to a decrease in the proliferation ability of neural stem cells, and inhibited the differentiation of neural stem cells into immature neurons and end-stage astrocytes. Conclusion Ifitm3 knockout mice were established using CRISPR/Cas9 method, and neural stem cells were isolated and cultured in vitro, which proved that the lack of Ifitm3 had the effect on the self-renewal, proliferation and differentiation of neural stem cells, providing insights into the biological function of IFITM3.
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Dynamic changes of MAOA during the neuroendocrine differentiation of prostate cancer

HAN Hao, LI Hui, ZHANG Caiqin, ZHAO Jumei, WEI Xiaoli, SHI Changhong

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Objective Exploring the dynamic changes of monoamine oxidase A (MAOA) and forkhead box A1 (FOXA1) during neuroendocrine differentiation (NED) in prostate cancer, providing new strategies for the treatment of neuroendocrine prostate cancer (NEPC). Methods Cell models and mouse transplantation models of NED were established through long-term sustained induction of enzalutamide (ENZ); Dynamic expression of MAOA, FOXA1 in NED detected by Western blot and qPCR methods; Selection of GEO database to analyse the dynamic trends of MAOA and FOXA1 in multiple NED models; Construction of mouse transplantation model of human prostate cancer cell lines and analysis of the dynamic expression of MAOA and FOXA1 in NED in the in vivo model by immunohistochemistry; Intervention of MAOA by lentiviral transfection and detection of MAOA's regulation of FOXA1. Results Both MAOA and FOXA1 showed the dynamic characteristics of increasing and then decreasing during NED process; Knockdown of MAOA in prostate cancer cells can lead to decreased expression of FOXA1, which may be the reason why MAOA plays different roles through FOXA1 at different stages of NED.Objective To establish cellular and animal models of neuroendocrine transdifferentiation (NED) in prostate cancer, to further explore the role and mechanism of monoamine oxidase A (MAOA) in the process of NED. Methods The human prostate cancer cell line C4-2 was selected to induce the NED process by long-term stimulation of enzalutamide (ENZ); The changes in the levels of MAOA and Forkhead Box A1 (FOXA1) during the NED process were detected by Real-time PCR and Western blot; and a xenograft model of prostate cancer was established to validate the changes in MAOA and FOXA1 in vivo. The expression of MAOA and FOXA1 was further analyzed by utilizing GEO database sequencing data GSE8702 and GSE59986. Results ENZ can be used as a method to induce NED cellular model and animal model; during the process of ENZ-induced NED, the expression of MAOA and FOXA1 showed a dynamic change character, firstl increasing and then decreasing; Knocking down MAOA expression in prostate cancer can lead to the decrease of FOXA1 expression, which may be the various roles of MAOA through regulation of FOXA1 at different stages of NED. Conclusions Both MAOA and FOXA1 showed a trend of increasing and then decreasing during NED, and the expression of MAOA could affect the level of FOXA1, and MAOA/FOXA1 may play a dynamic regulatory role in the NED process.
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Research Progress of animal models of audiogenic seizures

ZHAI Wenji, WU Jie, ZHENG Tihua, ZHENG Qingyin

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Audiogenic seizures (AGS) is an epilepsy caused by strong acoustic stimulation with generalized muscle spasms during seizures, and its model is of great significance in the study of epileptogenesis, the search for causative genes and regulatory channels, and the screening of new antiepileptic drugs(AEDs). This paper summarizes the current progress of research on common animal models of AGS in terms of pathogenetic features, possible pathogenesis and causative genes to provide new research directions and targets for the development of AEDs.
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Using allostatic load as a tool for evaluating aging and the aging-delaying effects of Zuogui Pill

jiayuxin, dengxiaohong, sunleifeng, chendandan, duanxiaohong, yanshikai, huangjianhua

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Objective Using the concept and method of allostatic load (AL) to evaluate aging and the effectiveness of Zuogui Pill intervention in aging. Methods Natural aging rats were tested at the ages of 2, 5, 8, 14, and 18 months, and were divided into an elderly control group, a low-dose group of Zuogui Pill, and a high-dose group of Zuogui Pill. Intervention with Zuogui Pill for 3 months. Blood samples were taken from the tail of rats of each month and group, and the number of T lymphocytes and apoptosis rate were measured by flow cytometry. Low-density lipoprotein cholesterol (LDL-c), High-density lipoprotein cholesterol (HDL-c), triglycerides (TG), total cholesterol (tCHO), free fatty acids (FFA), 25 hydroxyvitamin D (25-OHD), corticosterone (CORT), C-reactive protein (CRP), Interleukin-6 (IL-6) in rat serum were detected. By identifying the collinearity between indicators and professional considerations, LDL-c, tCHO, HDL-c, FFA, TG, CORT, IL-6, CRP, 25-OHD, CD3+ T cell count, and CD3+ T cell apoptosis rate were included for AL scoring. The threshold for each indicator was established on data from 5-month-old rats, and the score was 1 point below or/and above the threshold. Results The serum LDL-c, TG, tCHO, 25-OHD, CRP, and IL-6 of rats showed significant changes with age, but the change patterns were different. The CD3+ T lymphocyte count significantly decreased with age (P < 0.01), while the apoptosis rates of CD3+, CD4+, and CD8+ T lymphocytes significantly increased with age (P < 0.01). Zuogui Pill can significantly increase the serum CORT level in elderly rats (P < 0.01) and reduce the apoptosis rate of CD8+ T lymphocytes (P < 0.05). The AL score began to increase at 5 months of age and reached its peak at 18 months of age. Conclusion AL can better characterize the aging process compared to a single indicator. Zuogui Pill can improve the stress response ability of aging rats and alleviate their immune aging.
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The Impact of Maternal Stress during Pregnancy on Placental Glucose Transporters

songqi, du zhixin, yang liping, houjunlin, zhaojiajia, wuyongye, wangyaohui, lixiaolin

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Objective Investigate the effect of fear stress during pregnancy on the expression of glucose transporter (GLUT) in the placenta, providing evidence for the theory of fetal damage caused by fear stress during pregnancy. Methods 20 pregnant Wistar rats were randomly divided into a control group and a model group, with 10 rats in each group. Among them, the fear stress model was established according to modified bystander electroshock method for 19 days, and the normal group was kept normally. After the experiment, the number of offspring, placental and fetal rats weights were measured, and placental efficiency was calculated. Transmission electron microscopy was used to observe the morphological changes of placental cells. Bioinformatics analysis was performed to screen for differential genes in pregnancy stress-phobic placentas, and GSEA enrichment analysis was performed. Protein immunoblotting (Western blot), real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and immunohistochemistry were used to detect GLUT1, GLUT3, GLUT6, and GLUT7 protein and gene expression levels. Results The placental efficiency was significantly reduced in the model group compared with the control group; the results of transmission electron microscopy showed that the placental microvilli were sparse and short, and the mitochondria and endoplasmic reticulum were swollen in the model group; GSEA analysis revealed that placental genes were significantly enriched in cellular glucose homeostasis in the model group compared with the control group; The results of Western blot, Real-time PCR and immunohistochemistry indicate a decrease in both protein and gene expression levels of GLUT1, GLUT6, and GLUT7 in the placenta of pregnant rats. Conclusions Prenatal exposure to fear stress may lead to adverse pregnancy outcomes, potentially associated with reduced levels of three key glucose transporters in the placenta: GLUT1, GLUT6, and GLUT7.
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Research progress on the application of zebrafish model in Alzheimer"s disease

manhaishuo, chenhongli, heyue, sunyaxuan, huoqing, daixueling

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【】 Alzheimer"s disease (AD) is a degenerative disease of the central nervous system, which is mainly manifested by cognitive dysfunction and speech loss, and its pathogenesis is associated to many factors. In recent years, zebrafish has attracted extensive attention due to its high homology with human in brain structure and function, nerve conduction and AD pathogenic genes. In this paper, we reviewed the advantages of zebrafish as an animal model to explore the pathogenesis of AD, AD drug screening, and drug evaluation, to provide new insights for the pathogenesis of AD and new drug development.
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Advances in animal models of intestinal fibrosis

wangxi, sunxin, zoudefang, gurenjun

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Intestinal fibrosis is a complication of inflammatory bowel disease, and its refractory and recurrent nature impose a serious disease burden on patients. The disease’s pathogenes is not clear, and there is no effective treatment. Moreover, there is still a lack of recognized intestinal fibrosis models. In this paper, we review the methods used to establish animal models of intestinal fibrosis both at home and abroad, and consider the clinical relevance, key characteristics, and advantages and disadvantages of the procedures. Intestinal fibrosis models were summarized according to the modeling period and methods.
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Application of Micro-CT in experimental animal disease model

LI Shuzhen, DAI Wenjing, YU Qingqing, TIAN Miao, ZHANG Qian, LI Bei

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Micro-CT, as a non-invasive technology, is widely used in animal experiments to assist in the detection of bone diseases, lung diseases, oral diseases, metabolic diseases, middle and inner ear diseases, tumors and other animal disease models, providing diverse, scientific and reliable image data for animal experiments. It has become one of the indispensable experimental methods in animal experiments. This review will introduce the imaging principles of Micro-CT, review its application in the study of animal disease models, summarize the limitations of Micro-CT technology, and look forward to the future prospects.
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Methodological study on constructing a mouse model of bile stasis caused by partial bile duct ligation

TU Haiye, BAO Fangqi, ZHANG Lizong, JIANG Chen, WEN Sisi, ZHAO Ziyu, FANG Mingsun, CHEN Minli

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【Abstract】 Objective To observe the effects of different ligation sites and fasting methods on a C57BL/6J mouse model of partial bile duct ligation (pBDL) induced cholestasis, and study a pBDL modeling method with high modeling rate, typical symptoms, and good stability. Methods C57BL/6J mice were subjected to left bile duct ligation (L-pBDL) and left to median bile duct junction ligation (ML pBDL) for modeling, and the effects of different pBDL ligation methods on serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin (TBIL), total bile acid (TBA), and liver histopathology of the model were observed; And the effects of different fasting methods on the symptoms and liver injury of the ML-pBDL model were observed by fasting for 12 and 16 hours before surgery, and fasting and fasting for 4 hours after surgery. Results (1) The incidence of jaundice in the ML-pBDL group was 52.94%, and the survival rate within 3 weeks after surgery was 64.71%, while the incidence of jaundice in the L-pBDL group was 11.76%, and the survival rate within 3 weeks after surgery was 82.35%; Compared with the sham surgery group, the serum liver function indicators in the L-pBDL group and the ML-pBDL group were significantly increased (P<0.01), and the ALP activity in the ML-pBDL group was significantly higher than that in the L-pBDL group (P<0.05); Compared to the L-pBDL group, the ML-pBDL group had more severe liver fibrosis at 3 weeks post surgery (P<0.01). (2) The incidence of jaundice in the 16 hour fasting group was 93.33%, and the survival rate within 3 weeks after surgery was 73.77%. However, the incidence of jaundice in the 12 hour fasting group was 42.86%, and the survival rate within 3 weeks after surgery was 71.42%; Compared with the normal group, the ALP activity, ALT/AST ratio, TBA level, and proportion of collagen fiber area were significantly increased in the 16 hour and 12 hour fasting groups (P<0.05) The observed indicators in the 16 hour fasting group were higher than those in the 12 hour fasting group, but there was no significant difference (P>0.05). Both the 12 hour and 16 hour fasting groups showed significant bile duct hyperplasia and liver fibrosis (P<0.01), and the liver fibrosis in the 16 hour fasting group were more severe (P<0.01). Conclusion Both L-pBDL and ML-pBDL ligation methods can establish a mouse model of cholestasis. However, the symptoms of the L-pBDL model only exhibit transient damage characteristics, while the liver lesions of the ML-pBDL model are typical and stable. Prolonging preoperative fasting time can improve the modeling rate and stability of the ML-pBDL model, and the pathological symptoms are more typical.
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Research Progress on Osteoporosis Combined with Animal Models

LI Yongjin, ZHANG Xiaoyun, ZENG Hao

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With the intensification of aging, the incidence of osteoporosis (OP) has increased year by year and become one of the global public health problems. Its main features are decreased bone mass and destruction of bone microstructure, and clinically it often presents with pain, kyphosis, decreased height, and even fractures. At present, the clinical treatment methods are mainly Western medicine, but there are many drawbacks, and traditional Chinese medicine has the advantages of low price and small toxic side effects in the process of prevention and treatment of OP. As an important means in the basic research process of traditional Chinese medicine, the construction of animal models plays a very important role in the process of medical development, and the combination of disease evidence and animal models has become one of the indispensable methods for the modern research and development of traditional Chinese medicine in China. The purpose of this paper is to summarize the model construction and evaluation methods based on the current research status of disease evidence combined with OP animal models, in order to provide certain ideas and references for the research of disease evidence combined with OP animal models.
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Progress on mitochondrial injury in mdx mice

gaotian

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【Abstract】Duchenne muscular dystrophy (DMD) is a lethal, progressive, X-linked recessive hereditary muscle disease caused by a mutation in the gene encoding dystrophin. Currently, no cure for DMD is available, and clinical research is progressing slowly. The establishment of animal models is becoming increasingly important for experimental research on DMD. Following the research findings that mdx mice have the same pathogenesis as DMD patients, this model is widely used in the study of DMD pathogenesis and drug development. Mitochondrial injury is one of the most important pathological mechanisms of DMD, and mitochondrial protection is a potential therapeutic strategy for DMD, and thus it is significant to study mitochondrial injury in mdx mice. This article reviews the progress of research into mitochondrial injury in DMD model mdx mice in recent years to provide a reference for related experiments.
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Prevalence, prevention and control of tuberculosis in monkey

zhaiwei, LiuDonghui, Xu Zhengzhong, Zheng Chengkun, Jiao Xinan, Chen Xiang

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Non human primates (NHPs) are susceptible hosts of tuberculosis(TB). After infection, TB will not only spread among populations, but also spread to humans. Currently, there is no effective vaccine to prevent NHPs from TB. Although current prevention and control protocols are more mature and have reduced the incidence of NHPs TB in captivity, outbreaks continue to occur. This article summarized the epidemiological situation of TB in monkeys in captivity and wild environment around the world, analyzed the advantages and disadvantages of the current common detection methods, and summarized the most common practice of the current prevention and control of NHPs, indicating that TB poses a great threat to NHPs, so as to improve the awareness of TB among NHPs breeding workers and managers, provide a basis for improving the current management procedures, and provide a reference for the diagnosis, prevention and control of monkey tuberculosis in China.
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Study on the sweat secretion and the biochemical indexes in Yin-deficiency ovariectomized rats

Wang Jie, Chen Wenjia, Lin Wenwu, Wang Yingzheng, Wang Yinghao, Huang Meixia

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Objective To assess the sweat secretion of Yin-deficiency ovariectomized rats and to investigate the changes of biochemical indexes. Methods Eighteen SD female rats were randomly divided into sham operation group, model group, and positive control group with six rats in each group. The sham operation group were taken sham operation, the model group and the positive control group underwent bilateral ovary removal surgery, and L-thyroxine (92mg·kg-1) was given once a day from the 7th day after the operation for 7 consecutive days to establish Yin-deficiency ovariectomized model. The positive control group was orally administered Qinggu San Tang (7.3g·kg-1) daily, while the sham operation group and model group were orally administered an equal amount of distilled water once a day for a total of 14 days. Sweat secretion from the rat foot-plantar region was determined using the Wada-Takagaki reagent colouring method. At the end of the experiment,blood was taken from the abdominal aorta, and the tissue of the paw pads of rats were separated. Serum cAMP, cGMP, LH, GnRH, and E2 levels were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was used to determine the expression levels of M3 receptor, β2 receptor, and aquaporin-5(AQP5) in the paw pad. Results (1) Compared with the sham operation group, rats in the model group were more irritable and aggressive, their body weights decreased while their average temperature increased significantly, sweat secretion increased significantly; (2) Serum cAMP and cAMP/cGMP ratio increased, LH and GnRH increased significantly, and E2 level decreased; (3) M3 receptor expression was down-regulated and β2 receptor, aquaporin (AQP5) expression up-regulated in paw pads of rats. After 2 weeks of positive control treatment, serum cAMP and the ratio of cAMP/cGMP decreased significantly, LH and GnRH levels decreased, but no statistical difference was observed for serum E2 level. The expression levels of M3 receptor, β2 receptor, and aquaporin (AQP5) were down-regulated in paw pads of rats. Conclusion: Sweat secretion was significantly increased in the perimenopausal syndrome kidney yin deficiency "combined disease and evidence" model established by desiccation combined with thyroxine, and the mechanism may be related to the changes of cGMP, cAMP, and its key proteins M3R, β2AR, and AQP5 in sweat glands that regulate sweat secretion.
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Comparison and optimization of special staining methods for myocardial fibrosis observation

wang ya heng, ma jia xin, lei yu, zhang lian feng, lv dan

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Objective Myocardial histopathological detection is the gold standard for the diagnosis of myocardial fibrosis, and special staining is a commonly used detection method in pathological examination.However, at present, there are some problems in the software analysis of commonly used special staining methods for myocardial fibers, such as the difficulty in identifying the location of collagen and the large error in quantitative analysis, and it is of great clinical significance to explore the best special staining method for the diagnosis of myocardial fibrosis. Methods Paraffin sections of cardiac tissue were prepared using a transgenic mouse model of cardiomyopathy with specific cTnT R141W gene mutation caused by myocardial tissue mutation constructed in the laboratory, and four staining methods were performed, includind Masson's trichrome staining (Masson), Picric Sirius red staining (PSR), van Gieson staining (VG), and Sirius red/Fast Green staining(SR/FG), and comparative observation. Subsequently, Image J software was used to quantitatively compare the area of collagen fibers.The Sirius red/Fast green staining method was optimized from three aspects: dye concentration, staining time, and acid solution prestaining, and the subsequent quantitative analysis of collagen fibers was applied and verified. Results The distribution of collagen fibers could be observed by the four special staining methods, among which the Sirius red/Fast green staining method, 0.1% Sirius red-picric acid acidic solution was used to pre-stain for 5 min, and the concentration of the dye solution was adjusted to 0.1% Sirius red-picric acid and 0.04% fast green mixture, and the sections were placed in the mixed staining solution for 1h, has the lowest missed reading and loss in determine the proportion of collagen fibers. Conclusion Compared with other traditional collagen fiber special staining methods, the optimized Sirius red/Fast green staining method in this paper has bright coloring between collagen fibers and myocardial tissue, obvious color contrast, good stability, convenience and speed, and is more suitable for subsequent quantitative analysis and application of collagen fiber proportion.
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Construction of immortalized corneal stromal cell line of tree shrew and study on viral infectivity

DING Xiangrong#, CHEN Liu、#, HUO Shurui, QI Mengdi, LIU Xin, WANG Wenguang, LI Na, DAI Jiejie, LU Caixia

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Objective To establish an immortalized tree shrew corneal stromal cell line and to study the virus infection. Methods The primary corneal stromal cells (CSCs) of tree shrew were isolated and cultured by tissue block adhesion method. CSCs were transfected a lentivirus carrying SV40T gene, monoclonal clones were selected for passage culture. The characteristics of CSCswere tested through morphological observation、immunofluorescence、Karyotypeand cell proliferation curve. Using Herpes simplex virus-1(HSV-1)、ZIKV(GZ01 strain)、Dengue virus typeⅡand H1N1 A/Puerto Rico/8/1934(A/PR/8)to infect the CSCs. Virus titers were detected by CCDI50. Results the immortalized tree shrews CSCs transmitted to more than 50 passages were spindle-shaped and had good cell morphology and structure. High level of Vimentin and SV40T were detected by immunofluorescence. The cell growth curve showed the cells were in the logarithmic phase on days 4-5, and grew vigorously. The number of chromosomes of the primary cell karyotype was stable at62, while immortalized CSCs was 64 at P21 and P56. The results of virus titer showed that the immortalized tree shrew CSCs were sensitive to HSV-1、ZIKV(GZ01 strain)、Dengue virus typeⅡand H1N1 A/Puerto Rico/8/1934(A/PR/8),and the virus titers were 1.32×105^TCID50/mL、5.62×106^TCID50/mL、2.69×107^TCID50/mL、7.76×104^TCID50/mL respectively. Conclusions The immortalized tree shrew CSCs were successfully established, suggesting that the cell line can be used in study of the mechanism of herpes simplex virus, Zika virus, Dengue virus and influenza A virus infection of corneal diseases and antiviral drugs.
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Establishment of a genetically diverse hypertensive mouse model and analysis of gene transcription regulation

huangzhibin, panjirong, zhanglingyan, zhaodalu, wangqian, weichengzhi, maxu, bailin, qinchuan

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Objective To investigate the differences in blood pressure phenotype, renal pathological changes and related pathogenic pathways in genetically diverse hypertensive mice from 13 strains. Methods The mice with different genotypes were obtained by crossing 13 genetically diverse strains with Cckbr-/- mice, and then the blood pressure was measured by BP-2000 noninvasive blood pressure analysis system, the expression of CCKBR protein in mice kidney was detected by Western blot, the pathological changes of mice kidney tissue were detected by HE staining and immunohistochemistry (IHC), and the pathogenic pathways related to essential hypertension were screened by RNA-Seq. Results In 3 specific mouse strains (A/J, LOT, FIM), the systolic blood pressure (SBP) of Cckbr-/- group was significantly different from that of Cckbr+/+ group. HE staining and immunohistochemistry showed that hypertension caused a certain degree of renal injury in mice. GO and Pathway enrichment analysis showed that differentially expressed genes were enriched in metabolism and circadian rhythm regulation. Conclusion Genetically diverse mice can better simulate the genetic background of the population and provide a new resource for studying the pathogenic genes related to essential hypertension.
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Establishment and evaluation study of non-obese polycystic ovary syndrome rat model

Qiao Shiqing, Wang Ting, Yan Yonghuang, Yang Jiusi, Yu Yuling, Wang Yanmin, Sun Yateng, Wu Yujie, Zhu Peixuan, Li Min, Chen Cong, Su Zeqi, Zhang Cai

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Objective To establish a stable rat model of non-obese polycystic ovary syndrome (PCOS) with clinical characteristics. Methods The dehydroepiandrosterone (DHEA) was used to establish a PCOS rats model by subcutaneous injection. 3-week-old SD rats were divided into a normal group, a 6 mg/kg DHEA model group, and a 60 mg/kg DHEA model group, and the model groups were subcutaneously injected with DHEA according to the corresponding dose daily, while the normal group was subcutaneously injected with glycerol daily for 21 consecutive days. The model was evaluated with ovarian histopathology as the gold standard to determine the optimal dosage of DHEA-induced PCOS rat model. On this basis, the optimal DHEA modeling dose was selected, and the stopping and continuing modeling groups were set up respectively, to observe the model for 28 days to investigate maintenance of model. The stopping modeling group was no longer given DHEA, and the continuing modeling group was subcutaneously injected with 60 mg/kg DHEA every 48 h. The evaluation indicators include body mass, estrous cycle, fasting blood glucose and serum insulin, histopathologic morphology of the ovaries and serum sex hormone levels. Results 1) Compared with the normal group, body mass in the 6 mg/kg DHEA model group and the 60 mg/kg DHEA model group showed no significant difference, and the estrous cycles were irregular in both groups. And there were more cystically dilated large follicles in theovaries, with fewer mature follicles, reduced layers of granulosa cells in a sparse and disorganized manner, and fewer luteum in the 6 mg/kg DHEA model group and the 60 mg/kg DHEA model group. Furthermore, serum T and E₂ levels were significantly higher in the 60 mg/kg DHEA model group (P<0.05). 2) The stopping modeling group: the model group resumed regular estrous cycles after two weeks compared with the normal group, various growth follicles and corpora lutea were observed in ovarian tissues, the number of cystic follicles were reduced, the number of granulosa cell layers was increased, mature follicles were visible, and oocytes were locally intact in morphology, furthermore, the levels of E₂ and AMH were reduced in the model group (P<0.05). 3) The continuing model group: Compared with the normal group, the model group was in the late stage of estrous cycle for a long period, and there were more large follicles with cystic dilatation, fewer mature follicles, fewer layers of granulosa cells with sparse and disordered arrangement, the number of corpus luteum was significantly reduced in the ovarian. Furthermore, the levels of serum LH , LH/FSH and T were elevated (P<0.05). Conclusion Subcutaneous injection of 60 mg/kg DHEA for 21 consecutive days can successfully construct a non-obese PCOS rat model that meets clinical characteristics. On this basis, subcutaneous injection of 60 mg/kg DHEA every 48 hours can maintain the stability of the model.
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Network pharmacological study and experimental validation of Rhizoma Chuanxiong volatile oil in the treatment of angina pectoris

YangMing, WangChaoping, LeiZhiqiang, LiuBo, ZhangHua, LuoJing

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Objective Based on network pharmacology and animal experiments, this study aims to study and verify the therapeutic effect of the volatile oil of Rhizoma Chuanxiong on angina pectoris. Methods Volatile oil components were screened out by steam distillation, GC-MS and ADME parameters. Using Pubchem, SwissTarget, DisGENET, DrugBank database and R language to obtain the target of volatile oil components, angina pectoris target and intersection target respectively; Protein-protein interaction is completed through the String database; The clusterProfiler package of R language was used to analyze the GO and KEGG enrichment of the intersecting targets and construction of herbal-component-target-pathway networks by Cytoscape. Molecular docking using AutoDock vina, Pymol and Discovery Studio 2016 software, molecular docking was carried out to analyze the affinity between key targets and main volatile oil components. Finally, the therapeutic effect of volatile oil of Rhizoma Chuanxiong on angina pectoris was further verified by animal experiments. Results There were 10 volatile oil components and 22 key targets, which were closely related to neurotransmitters, receptors on synaptic membranes, material metabolism, neuroactive ligand-receptor interaction, retinol metabolism, drug metabolism-cytochrome P450 pathways. Molecular docking results showed that 3-butylidenephthalide, Alpha-selinene, Trans-ligustilide and other volatile oil components combined with several key targets play a therapeutic role. Animal experiments show that the volatile oil of Rhizoma Chuanxiong can regulate the levels of EF, FS, SV, LVIDs and the activities of LDH, CK and AST; and promote the expression of ADRA1A and CHRM5 proteins in damaged cardiomyocytes; improve the state of myocardial fibers, reduce intercellular space, and reduce inflammatory cells Infiltration appears to protect the myocardium. Conclusion In summary, the volatile oil of Ligusticum wallichii can effectively protect the damaged myocardial tissue and play a role in treating angina pectoris.
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Establishment and optimization of C57BL/6J mice liver fibrosis model induced by carbon tetrachloride

Sun jingran, Lu Bingjiu, Zheng Jialian, Sun Xiaoning, Xu Junchao

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Objective To optimize C57BL/6J mouse liver fibrosis model induced by different doses of carbon tetrachloride through imaging, molecular biology and pathology methods.Methods Thirty-six healthy C57BL/6J male mice were randomly divided into control group, 2 weeks group, 3 weeks group, 4 weeks group, 6 weeks group and 8 weeks group (n=6) after adaptive feeding for 1 week. The control group was intraperitoneally injected with 0.2 mL olive oil injection three times a week, and the positive control groups were intraperitoneally injected with 0.2 mL 20% CCl4-olive oil solution three times a week. The changes in body weight of mice in each group were recorded. Liver stiffness was measured on days 15, 22, 29, 43 and 57, and blood samples were collected, the mice were measured cereal third transaminase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), layer mucins (LN), type Ⅲ before collagen (PC-Ⅲ) and collagen type Ⅳ (Ⅳ-C) content, The liver tissues were stained with HE, Masson and Sirius red.The Metavir scoring system was used to evaluate the degree of liver fibrosis.Results Compared with the control group, the mice in the positive control groups were listless and clustered together. In terms of body weight, the 4-week group, 6-week group and 8-week group were significantly lower than the control group, while the 2-week group was significantly higher than the control group. Liver elastography showed a progressive increase in stiffness with the increase of administration time. Biochemical results show that, compared with the control group, the other groups ALT, AST levels were significantly higher, with the increase of drug delivery time, the positive control group HA, LN, PC-Ⅲ and Ⅳ-C levels showed a trend of rise. The pathological results showed that the degree of liver fibrosis was progressively aggravated with the increase of administration time. At 4 weeks, the pathological diagnosis of liver fibrosis was consistent with that of liver fibrosis, there was a trend of pseudolobules formation at 6 weeks, and pseudolobules were formed at 8 weeks, suggesting early cirrhosis.Conclusions The liver fibrosis model of C57BL/6J mice can be successfully established by intraperitoneal injection of 20% CCl4-olive oil solution three times a week for 4 consecutive weeks, which has good stability and rapid modeling. It can be used as an optimized scheme for the establishment of liver fibrosis model.
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To explore the mechanism of Xuanfei Jiedu Formula against multidrug-resistant Pseudomonas aeruginosa pneumonia based on TLR4/Myd88/NF-κB signaling pathway

SHEN Ting ting, LI Ya, LI Su yun, LI Gao feng, HAN Bin Yang

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【Abstract】Objective To investigate the mechanism of Xuanfei Jiedu Formula on multi-drug resistant Pseudomonas aeruginosa pneumonia. Methods In addition to the blank control group, the other groups used tracheal intubation to establish a rat model of MDR-PA (9×108 CFU/ML, 0.5 mL) pneumonia, and were randomly divided into the model group, Xuanfei Jiedu formula low dose group, Xuanfei Jiedu formula medium dose group, Xuanfei Jiedu formula high dose group, and Imipenem cilastatin group. After 1 day of modeling, 1.725g/kg/d, 3.45g/kg/d and 6.9g/kg/d of XFJDF were given by gavage in the low, medium and high dose groups respectively, 410 mg/kg/d of IPM was given by intraperitoneal injection in the imipenem cistatin group, saline was given by gavage in the blank control group and the model group; 2 times/day for 7 days. We observed the state changes, body weight changes, and wet-to-dry weight ratio (W/D) of the lung tissues of the rats, the histopathological changes of the lung tissues in each group under the light microscope using Hematoxylin-Eosin staining (HE) , detected the serum of the rats in each group by using enzyme immunosorbent assay (ELISA) to detect the interleukin-1β, interleukin-6, tumor necrosis factor-α (TNF-α) inflammatory factors , the content of reduced glutathione (GSH) and the activity of myeloperoxidase (MPO) were detected by colorimetric assay, the content of myeloperoxidase (MPO) was detected by TBA.The localization and semi-quantitative observation of TLR4, Myd88, and NF-κB p65 proteins in lung tissues were carried out by immunohistochemistry (IHC).Real-time fluorescence quantitative PCR (RT-qPCR) and protein immunoblotting (Western Blot) were used to detect the expression levels of TLR4, Myd88, NF-κB mRNA and protein in the lung tissues of rats in each group. Results Compared with the Control group, rats in the Model group showed delayed response, increased respiratory rate and murmur, different degrees of chills, decreased diet and water intake, weight loss; the W/D of the lung tissue (P<0.01) was significantly higher, and the alveolar cavities and peribronchioles of the lung tissue contained a large number of inflammatory cell infiltration, some of the alveolar walls showed fracture and fusion to form an airspace with inflammatory exudation, while the interstitial spaces of the lung tissue showed a large number of inflammatory cell infiltration. Inflammatory exudation, interstitial thickening and localized pulmonary fibrosis were observed. Serum levels of IL-1β, TNF-α and TGF-β were significantly elevated (P < 0.01) and the levels of MDA, MPO and GSH were increased (P<0.01) the expression of mRNA and protein of TLR4, Myd88, and NF-κB was significantly elevated (P<0.01) in the lungs. Compared with the Model group, all the treatment groups of the intervention group improved the general state of MDR-PA rats, which showed significantly better mental state, enhanced response sensitivity, increased body weight, decreased lung W/D (P<0.01), significantly improved pathological injury of lung tissues, significantly decreased levels of TNF-α, TGF-β and MDA, increased levels of GSH in serum (P < 0.01); except for the Except for the XFJDF low-dose group, the serum levels of IL-1β and MPO activity were significantly reduced (P<0.01); the mRNA and protein expression levels of TLR4, Myd88 and NF-κB were significantly reduced in the lung tissues (P< 0.01, P< 0.05), the most significant reduction was observed in the XFJDF high-dose group and the IPM group. Conclusion Xuanfei Jiedu formula significantly improved the general status, body weight, lung W/D and lung histopathology, reduced inflammation and oxidative stress in MDR-PA rats, its mechanism may be related to the inhibition of the TLR4/Myd88/NF-κB pathway expression in the lungs by Xuanfei Jiedu formula.
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Advances in the study of neural mechanisms in animal models of anger

ZHANG Chunpeng, GAN JIA HONG, ZHANG Yonghua

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The negative emotion of anger can have many effects on the body. The brain regions associated with the production of anger are mainly related to the central gray matter, amygdala, and hypothalamus. Animal models are an important tool for understanding diseases and mechanisms, and in recent years there has been a gradual increase in research on animal models of anger and the mechanisms of anger. Most studies on anger models have focused on the mid-suture dorsal nucleus, hypothalamus, and hippocampal regions, and related neurotransmitter studies have mainly been related to GABA expression and monoamine neurotransmitter content. The aim of this paper is to summarize the intracerebral neural mechanisms of anger onset by retrieving anger-related animal models, hoping to provide useful references for the study of angry emotions.
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Exploration of establishing the dynamic time model of acute lung injury in mice based on the NLRP3/caspase-1/GSDMD signal pyroptosis pathway

Fan Yixuan, Yang Zhixu

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Objective: Based on the NLRP3/Caspase-1/GSDMD pyroptosis pathway, a mouse model of LPS-induced acute lung injury was established over time, and the lung injury of mice was observed at different time points. According to the changes in injury at different time points and the expression of pyroptosis pathway-related proteins, the best time point for modeling was screened comprehensively, which laid the foundation for animal models for subsequent experiments. Methods: 54 six to eight weeks of SPF male BAL b/c mice were divided into nine groups randomly, which including control group and building time group (1, 3, 6, 12, 18, 24, 48 and 72 h group), body weight, lung tissue was detected in general and pathological observation and semi-quantitative score, lung index, lung water content of wet and dry weight ratio; The white blood cell count and the concentrations of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), IL-18 and BCA protein in bronchoalveolar lavage fluid (BALF) were detected. Using western blot method to detect the lung tissue of classic coke death pathways related NLRP3, pro - Caspase 1, Caspase 1, GSDMD protein expression. Results: According to the body weight statistics, the body weight of each group decreased, and the 24h and 48h groups showed the most significant weight loss. The results of gross observation and pathological examination of lung tissue showed that the injury of lung tissue in 24h to 72h was severe, and the difference between each group and Con group was statistically significant. Lung index, lung water content and wet/dry weight ratio were significantly increased at 24-72 hours. Alveolar lavage fluid white blood cells from 6 h after building began to rise, 48 h can reach a peak, 72 h all keep increasing; IL-18 in lavage fluid began to increase at 24 hours and continued to increase at 72 hours. TNF alpha, beta, IL - 6, IL - 1 inflammatory factors were keep the highest level at the time of 6 h, 48 h significantly reduced; A lavage in protein concentrations within 24 h, 48 h and 72 h compared with Con group were significantly increased; Found by western blot method and ALI model each time series jiao wu pathway proteins NLRP3, pro - Caspase 1, Caspase 1 and GSDMD protein expression were raised, 24 h - 72 - h group of channel protein expression enhanced significantly compared with control subjects. Conclusions: Comprehensive analysis of the experimental indicators, inflammatory factors and the expression of pathway proteins in WB in different time groups showed that the mechanism of pyroptosis was closely related to the occurrence and progression of ALI, and it moved with time. From the experimental results, it was found that the expression of pyroptosis pathway was the most obvious and the lung injury was the most serious in 24h-48h. This study provides a model reference and experimental basis for the subsequent study of the specific mechanism and intervention targets of ALI.
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Effects of cultured Mycelium Cordyceps Sinensis on carbon tetrachloride induced liver fibrosis mice by regulating AMPK/SirT1 signaling pathway

Yang Zhao, Yan Ruanyu, Wu Hongyu, Huang Kai, Shen Li, Tao Yanyan, Liu Chenghai, Peng Yuan

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【Abstract】 Objective To investigate the effects of cultured Mycelium Cordyceps Sinensis (CMCS) on AMPK/SirT1 signaling pathway in carbon tetrachloride(CCl4)-induced liver fibrosis in mice. Methods Forty male C57BL/6 mice of SPF grade were randomly divided into normal control group, CMCS control group (3.0g/kg/d), Model control group, CMCS1.5g/kg group(1.5g/kg/d) and CMCS 3.0g/kg group (3.0g/kg/d). Mice were intraperitoneally injected with 10% CCl4 (2ml/kg) to induce liver fibrosis. 2 weeks later, serum levels of ALT, AST and TBil were measured. Inflammation and collagen deposition in liver tissue were observed by H&E and Sirius red stainings, respectively. The content of Hyp in liver tissue was detected by Jamall's hydrochloric acid hydrolysis method. The levels of IL-6, MCP-1, IFN-γ, TNF, IL-10 and IL-12p70 in liver tissue were detected by CBA analysis system. Immunohistochemistry was used to observe the expression of Collagen I and SirT1 in liver tissue. Real-time fluorescent quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to observe the levels of Prkaa1, Prkaa2, Lkb1 and p53 in liver tissue. Results Compared with the normal control group, the serum levels of ALT, AST and TBil in the model control group were significantly increased(P<0.05). HE and Sirius red staining showed a large number of inflammatory cell infiltration and collagen deposition in the liver, respectively. The levels of Hyp content and the expressions of IL-6, MCP-1, and TNF in the liver tissues exhibited a significantly higher level (P<0.05). Conversely, the expressions of IL-10 and IL-12p70 displayed a decrease in significance (P<0.05). Immunohistochemical staining revealed an increase in the expression of Collagen I. And the staining of SirT1 was decreased in the hepatic sinusoidal space, while it was increased in the collagen deposition. RT-qPCR presented that the expression of Prkaa1, Prkaa2, and Lkb1 in liver tissue decreased, and the expression of p53 increased(P<0.05), respectively. CMCS could significantly reduce serum ALT and AST levels, decrease the expression of IL-6, MCP-1 and TNF in liver tissue(P<0.05), up-regulate the levels of IL-10 and IL-12p70(P<0.05), alleviate liver inflammation, collagen deposition and Hyp content, up-regulate the expression of SirT1 in the hepatic sinusoidal space, enhance the levels of Prkaa1, Prkaa2 and Lkb1(P<0.05), down-regulate collagen I and p53(P<0.05) in liver, respectively. In comparison with the CMCS 1.5g/kg group, the CMCS 3.0g/kg group exhibited a significant inhibitory effect on liver inflammation, collagen deposition and up-regulate AMPK/SirT1 expression(P<0.05). Conclusion CMCS could improve CCl4-induced liver fibrosis, and its mechanism was associated with the up-regulation of AMPK/SirT1 signaling pathway.
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Establishment of inhibition model of bone formation induced by lipid toxicity in zebrafish induced by palmitic acid

Wang Xiaoyi, LI Miao, WANG Linxia, YU Bin, HUA Yongqing

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Objective To establish a palmitic acid-induced bone formation inhibition model of zebrafish lipotoxic injury. Methods The embryos of AB strain zebrafish were randomly divided into blank Control group (Control group), palmitic acid group (PA group) and simvastatin group (SIM group). From 3 dpf (days post fertilization), PA and SIM groups were given PA . From 5 dpf, the SIM group was administered continuously for 4 days. Calcein staining, Nile red staining, triglyceride and total cholesterol content determination and q-PCR to determine whether the model was successfully established on the 9 dpf. Results PA significantly decreased the number of vertebrae, promoted lipid accumulation, increased triglyceride triglyceride and total cholesterol content, promoted lipid-related genes and inhibited the expression of osteogenic genes. SIM can improve the inhibitory effect of PA on bone formation in zebrafish. Conclusion PA can successfully create the inhibition model of lipotoxic injury bone formation similar to the pathological process of OP, and the method is simple, sensitive and controllable, which can be used for drug screening of OP and related diseases.
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Establishment and Evaluation of a Canine Vertebral Augmentation Puncture Model under Fluoroscopic Guidance

wanghaotian, liujia, huangjian, qijunqiang, xuguohua

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Objective: To establish a fluoroscopic percutaneous vertebral augmentation model in dogs through measurement and analysis of canine spinal anatomy. Furthermore, the effectiveness and safety of this modeling method are assessed through post-operative radiological analysis. Methods: Morphological measurements and their parameters of the lumbar vertebrae of six dogs aged around 12 months were taken: height of the L1-L7 vertebrae, width of the vertebral base, distance from the upper edge of the intervertebral disc to the narrowest part of the vertebra, distance from the vertical line of the spinous process to the upper edge of the intervertebral disc, and the vertical distance from the midpoint of the transverse process to the lower edge of the intervertebral disc. This was done to clarify the anatomical characteristics of the canine vertebrae and to determine the optimal location, direction, and depth for bone cement injection. Subsequently, a percutaneous vertebral augmentation model was established in the L4, L5, and L6 vertebrae of six healthy Beagle dogs weighing between 20-25kg. Four weeks post-surgery, the subjects were euthanized and radiologically examined. Primary observations included: surgical duration, post-operative distribution of the implanted bone cement, and the integrity of the vertebral canal and anterior edge of the vertebrae. Results: Anatomical observation of the canine vertebrae revealed that the vertebral height gradually increased from L1 to L5 and then decreased from L5 to L7. The width of the vertebral base consistently increased from L1 to L7. The distance from the vertical line of the spinous process to the upper edge of the intervertebral disc showed an increasing trend from L1 to L7 (1.9 to 4.0 mm). The distance between the midpoint of the base of the transverse process and the lower edge of the intervertebral disc gradually increased from L1 to L5 (4.7 to 6.9 mm). There was no significant statistical difference in the distance between the midpoint of the base of the transverse process and the lower edge of the intervertebral disc in the L4, L5, and L6 segments among the dogs (P=0.925). The midpoint of the root of the transverse process of the spine was taken as the puncture point, and the insertion direction and horizontal plane were at an Angle of 20° - 30 °, with a head tilt of 5° - 15° and a puncture depth of 1.2 cm to 1.5 cm. If the puncture was directed towards the caudal side of the vertebra, the angle of the needle tail was 30° to 35°, with a penetration depth of 1.5 cm to 1.8 cm. This technique allowed for the successful construction of a canine vertebral puncture surgical model. A total of 15 canine vertebral puncture surgical models were successfully created, with an average surgery time of 22.7 ± 4.6 minutes (ranging from 15 to 30 minutes) per vertebral segment. During surgery, one vertebral segment experienced spinal cord injury resulting in paralysis of the hind limbs and incontinence of both bowel and bladder. Two vertebral cortical bones fractured, but there were no deaths due to anesthesia or infection. Four weeks post-surgery, Micro-CT three-dimensional reconstructions consistently showed bone cement distributed within the trabecular bone of the canine vertebrae, with newly formed bone tissue enveloping the implanted material. There was no leakage, and no complications such as damage to the vertebral canal or the anterior wall of the vertebrae were observed. Conclusions: Based on the anatomy of the canine lumbar vertebrae (L4 to L6), using the midpoint of the base of the transverse process as a bony landmark, a safe and reliable canine vertebral augmentation puncture model can be successfully established.
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Differential expression analysis of mRNAs in the pneumoconiosis based on transcriptome sequencing

liang xiaoqiao, yang ying, he zhouyang, ning li

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【Abstract】Objective Utilizing transcriptomic sequencing, this study aimed to monitor the expression alterations of GIMAP8 and SEC14L5 throughout the progression of pulmonary fibrosis, thereby providing insights into the underlying mechanisms of its pathogenesis and evolution. Methods C57BL/6 male mice were assigned in a randomized manner to either the experimental or control cohorts. The model group underwent non-exposed endotracheal intubation on days 0 and 14 with 50 μL 100 mg/mL silica suspension, while the control group received 50 μL phosphate-buffered saline solution. On day 28, lung function was detected and the mice were sacrificed, and lung morphology, fibrosis, and mRNA levels were observed. Results The pneumoconiosis group was found to have a total of 2,988 mRNAs compared with the control group, of which 626 mRNAs showed large expression differences, including 242 up-regulated and 384 down-regulated genes. The enrichment analysis indicated that the primarily affected mRNAs with altered expression were associated with pathways such as p53, nuclear factor-κB, tumor necrosis factor, AMP-activated protein kinase, and other signaling pathways. In the model mice, the alveolar structures were compromised, characterized by the presence of collagen fiber accumulation and the formation of fibrous masses. In contrast, the control mice maintained a normal pulmonary architecture. GIMAP8 expression was up-regulated whereas SEC14L5 expression was down-regulated in lung tissues in the model mice, and mice in the model group had poorer lung function. Conclusions The onset and progression of pulmonary fibrosis may be significantly influenced by GIMAP8 and SEC14L5 expression in the blood in patients with pneumoconiosis and in silicosis animal models.
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Development of Microsatellite Markers and Analysis of Genetic Characteristics in Closed Colony of Apodemus peninsulaeDevelopment of Microsatellite Markers and Genetic Diversity Analysis of Apodemus peninsulae

ZHANG Qing, ZHANG Xiwen, HE Song, YUAN Bao, CHEN Jian, REN Wenzhi, QUAN Fushi, HU Jinping, DING Yu

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Objective The development of polymorphic microsatellite markers of A.peninsulae can enrich the genetic data of A.peninsulae and lay a foundation for genetic quality control and gene mapping of A.peninsulae.Based on genomic data, microsatellite loci of A.peninsulae were developed for genetic quality analysis of A.peninsulae and enriched the genetic data of A.peninsulae. Methods Microsatellite loci were screened based on the genome sequence of Apodemus peninsulae, and microsatellite primers were mined. The genetic diversity of the population were analyzed by multiplex PCR. Results Thirty microsatellite markers were successfully developed, and 30 microsatellite loci were evaluated using 60 samples of Apodemus peninsulae. A total of 152 alleles were detected, with an average number of observed alleles of 5.067. The average observed heterozygosity was 0.592. The average Shannon index was 1.2645 ; the average polymorphism information content ( PIC ) was 0.598. Conclusions Based on the microsatellite loci developed in this study, the genetic diversity of A.peninsulae population can be effectively analyzed, which lays a foundation for the establishment of genetic quality standards and genetic quality detection methods of A.peninsulae.
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Analysis of gut microbiota in a rat model of polycystic ovary syndrome induced by letrozole

YANG Hong, ZHANG Zhen, FU Xufeng

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Objective: To investigate whether letrozole (an estrogen synthesis inhibitor) can affect the composition, diversity and abundance of intestinal microbiota in patients with polycystic ovary syndrome (PCOS). Methods: The PCOS rat model was induced and constructed by letrozole, an estrogen synthesis inhibitor. The gut microbiota structure of normal and PCOS rats was analyzed by 16S rRNA sequencing. Results: At the phylum level, the PCOS group had a lower abundance of Firmicutes (77.2% vs. 73.3%) and a higher abundance of Bacteroidetes (17.2% vs. 23.9%) than the normal group. At the genus level, the abundance of Rochella (16.1% vs. 5.6%) and Zurichella (10.0% vs. 1.8%) in PCOS group was lower than that in the normal group. However, the abundance of Lactobacillus (15.9% vs. 18.8%)、 Prevotella (0.5% vs. 4.6%) and Ruminococcus (1.2% vs. 3.0%) increased. The KEGG signaling pathways of the two groups of differentially expressed bacteria were further analyzed. The results showed that the signaling pathways related to bile acid biosynthesis in the intestine were changed in the PCOS rat model. Conclusion: These results provide a basis for further in-depth study of microbial structure and function and regulatory mechanisms in PCOS patients.
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Study on the Micro-CT based animal model of lung adenoma in BALB/c mice

jian qin, xiang sirui, wang chuchu, chen wu, fu xi, you fengming, zheng chuan, lin junzhi

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Objective To establish an animal model of lung adenoma in BALB/c mice based on dynamic characterization by micro-computed tomography (CT). Methods Eighty female SPF-grade BALB/c mice were divided randomly into four groups: model low-dose group (1 mg/g urethane, intraperitoneal [ip], once), model medium-dose group (1 mg/g urethane, ip, once a week, followed by 2 weeks), model high-dose group (1 mg/g urethane, ip, once a week, followed by 4 weeks), and blank group (equal volume of saline). Growth of lung nodules in the mice was monitored regularly using micro-CT. Three-dimensional images of the lungs were drawn using the Analyze 12.0 system, and lung tissues were taken for histopathological examination (hematoxylin and eosin). Results Lung nodules with round high-density shadows were observed at week 11 in all model groups compared with the findings in the blank group. The rate of nodule formation increased with increasing modeling weeks, with rates of nodule formation in the model high-, medium-, and low-dose groups of 93.8%, 93.8%, and 87.5%, respectively, at week 21. Most mice had two to four, followed by one, and one to two nodules, respectively. The average maximum diameter of the lung nodules in the low-dose group was significantly higher than the diameters in the medium- and high-dose groups (P < 0.05), but there was no significant difference in lung nodule volume among the three groups. Regarding pathological type, hematoxylin and eosin staining revealed that the tumors in all the model groups were lung adenomas. Conclusions Lung adenomas were successfully induced in all urethane-dose groups of mice and growth of the lung nodules could be characterized by micro-CT. The rate of nodule formation was highest in the medium-dose group, which developed a moderate number of lung adenomas and provided a stable model, and was thus considered the most suitable model for the study of lung adenomas in mice.
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Mechanism of Jianpi Huatan Fang in Intervention of Lipid Metabolism in PCOS-IR Rats by Regulating FOXO1/PDK4 Signal Pathway

yangluyu, yuximing

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Objective: To investigate the effects of Jianpi Huatan recipe on lipid metabolism and expression of FOXO1 and PDK4 in rats with polycystic ovary syndrome (PCOS) and insulin resistance (IR) . Methods: Fifty female rats were randomly divided into a blank control group of 10 and a model group of 40. A PCOS-IR rat model was established using a high-fat diet (8 weeks) combined with letrozole (added at weeks 4-8), and the model was evaluated through HOMA-IR and ovarian tissue pathology observation. Thirty successfully modeled rats were randomly divided into a model control group, a Jianpi Huatan Fang group (11.07g/kg), and a Metformin group (0.2g/kg), with 10 rats in each group. Drug intervention was given for 4 weeks. Hematoxylin eosin (HE) staining was used to observe the histopathology changes of the ovary; Fasting blood glucose (FBG) and blood lipids (triglyceride TG, cholesterol (TC), high-density lipoprotein (HDL-C), low-density lipoprotein (LDL-C) were measured by blood biochemical analyzer; Enzyme linked immunosorbent assay (ELISA) was used to measure the levels of sex hormones follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T) and insulin (FINS); Immunoblotting (WB) and real-time fluorescence quantitative PCR were used to detect changes in the expression of FOXO1 and PDK4 in the ovaries. Results: Compared with the blank control group, the model control group showed significant weight gain, significant polycystic ovarian changes, and dysregulation of glucose and lipid metabolism (P<0.01). Compared with the model control group, the weight growth rate of rats in the Jianpi Huatan Fang group slowed down, and the development of follicles improved. Serum LH, T, LH/FSH, FBG, FINS, HOMA-IR, TC, TG, LDL-C decreased, E2 and FSH increased, while ovarian FOXO1, PDK4 mRNA and protein expression were down regulated (P<0.05, P<0.01); It can increase the serum HDL-C content, but there is no statistically significant difference between groups (P>0.05). Conclusion:Jianpi Huatan recipe can effectively regulate the secretion of sex hormones, improve the ovarian reproductive function and regulate glucose and lipid metabolism in obese PCOS-IR rats, which may play a role by inhibiting FOXO1/PDK4 pathway.
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Analysis of Henoch-Schonlein Purpura Nephritis Animal Model Based on Data Mining

ChengQi, DING Ying

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Objective: To investigate the modeling of Henoch-Sch?nlein purpura nephritis based on data mining, and to provide a reference for the preparation of a standardized Henoch-Sch?nlein purpura nephritis animal model. Methods: We searched the China HowNet, Wanfang, Weipu, China Biomedical Literature Database, and PubMed Chinese–English Database by computer to obtain studies of animal experiments relating to Henoch-Sch?nlein purpura nephritis in the past 20 years. The species, modeling methods, dosage, dosing cycle, modeling standards, and detection indexes were screened manually, and a database was established by using Microsoft Excel 2021 software for statistical analysis. The association rules of high-frequency indicators were analyzed using SPSS Modeler 18.0, and Cytoscape 3.6.1 was used to visually upgrade the association network diagram. Results: A total of 106 articles that met the inclusion criteria were summarized. Sprague Dawley rats and Kunming mice were the mostly commonly used animal models of Henoch-Sch?nlein purpura nephritis and most studies used drug-induced models. Bovine serum albumin (BSA) + lipopolysaccharide + carbon tetrachloride + castor oil, ovalbumin + Freund’s complete adjuvant, gliadin + Indian ink, and BSA + staphylococcal enterotoxin B were used to produce the animal models, generally with cycles of 5–14 weeks. The standard of modeling was skin purpura and increased numbers of urine red blood cells. Proteinuria, glomerular mesangial hyperplasia in kidney tissue, and immune complex mainly composed of immunoglobulin A (IgA) deposited in small blood vessels indicated successful modeling. There were 36 medical indexes, including 23 indexes related to the kidney and urine and nine indexes related to blood. Among these, 10 indexes, such as 24-h urine protein quantification, interleukin, renal pathology, urine red blood cell count, IgA, circulating immune complex and creatinine were used in ≥ 10% of cases. Cluster analysis of high-frequency indicators showed that the comprehensive evaluation model of 24-h urinary protein quantification + interleukin + renal pathology + urinary red blood cell count + IgA was mostly used. Conclusion: Most existing animal models of Henoch-Sch?nlein purpura nephritis have used male Sprague Dawley rats or female Kunming mice, and most models were induced by drugs. Among these, the method of stasis-heat syndrome combined with IgA nephropathy (disease-syndrome combination method) has the advantages of good repeatability and a high modeling rate, and may thus provide a reference for the selection of animal experimental models of Henoch-Sch?nlein purpura nephritis.
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Construction of macrophage-specific KLF2 gene knockout mice

MENG Xue, WANG Xinzhong, GAO Shuibo, WU Hong

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Objective To establish a macrophage-specific KLF2 gene knockout mouse model, and explore the regulatory effect of KLF2 on macrophage inflammatory response. Methods The KLF2flox/+mice were constructed using CRISPR/Cas9 gene editing technology. By breeding with Lyz2-Cre+/+ mice and screening genotypes through PCR, the target genotype mice were obtained, and the KLF2 knockout efficiency was verified using genotyping, qPCR and western blotting. BMDMs were Separated and cultivated, and the mRNA levels of inflammation-related factors in LPS-induced BMDMs were detected. Results A KLF2flox/flox/ Lyz2-Cre+ mouse model was established. The levels of KLF2 mRNA and protein in mouse bone marrow and BMDMs were significantly lower than those in the control group, while the expression of KLF2 in heart, liver, and kidney showed no significant changes compared to the control group. No significant differences in body weight, diet, drinking water and appearance were found between the two groups. Under the stimulation of LPS, the expression level of IL-6 mRNA in KLF2 deficient BMDMs was significantly lower than those in the control group, IL-1, iNOS, and CD86 mRNA were significantly higher than those in the control group. Conclusions The macrophage-specific knockout KLF2 mouse model was constructed, laying the foundation for further research on the regulatory effect and mechanism of macrophage KLF2 on clinical inflammatory-related diseases.
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The models of adenoviral transfection and hypoxia/reoxygenation-induced injury in AMCMs

LI Xiaoru, YAO Xinye, LIU Jia, ZHANG Xiaoyu, ZHANG Yiman, LAI Baochang, MA Qiang, WANG Yidong, TIAN Hongyan, YIN Qian

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Objective To build the models of viral transfection and hypoxia-reoxygenation induced cellular injury in AMCMs which are isolated by a non-Langendorff method. Methods AMCMs were isolated, extracted, sedimented and wall-plated by a non-Langendorff method. The morphology and survival rate of isolated cells was evaluated at 2 h, 24 h, 48 h and 72 h after wall-plating. The isolated AMCMs were infected with adenoviruses carrying RFP-expressing vector. Fluorescence photographs were taken at 36 h and 48 h post infection and used for calculation of transfection efficiency. The cells were cultured under hypoxic for 45 min and reoxygenated for 24 h. The cells were then stained with propidium iodide (PI) to verify the establishment of hypoxia-reoxygenation injury model. Results the survival rate of AMCMs at 2 h, 24 h and 48 h after plating were comparable, whereas this number at 72 h was significantly reduced. More than 80% of cells were transfected with adenovirus at 48 h. The hypoxia-reoxygenation treatment induced 42% of cells stained by PI, suggesting successful establishment of AMCMs injury model. Conclusions In this study, we developed a non-Langendorff method for quickly and easily isolating AMCMs with high cell viability. The isolated cells can be efficiently infected with adenovirus and are response to hypoxia-reoxygenation injury. Our findings provide a systematic method for isolating AMCMs as well as gene modification and hypoxia-reoxygenation injury in these cells.
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Regulation of tryptophanmetabolism instress-relatedgastrointestinaldisorders

chenjingqing, zhengjianhua, dongqiaoyan, zhangwensheng, maliansu, qiuyefeng

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Exposure to different types of stressors has been known to increase susceptibility to diseases and disrupting the body homoeostasis. It is necessary for healthy animals and humans to maintain the physiological functions under stressful conditions. Intestine, the largest immune and metabolicsite, is important for animal health. The integrity of mucosal barrier and function are fundamental to protect the health of the intestine. It is reported that stress has profound effects on the gastrointestinal tract including alterations in gut permeability，intestinal barrier and homeostasis. Tryptophan, a functional essential amino acid，that alters the gut microbiota, regulates intestine structural and functional changes，thus contributing to host physiology and metabolism. Tryptophan metabolism perturbations and its metabolites alterations in brain and intestinal tissues during stress suggest that tryptophan may play an important role in stress response. We therefore review the literature on the underlying mechanisms of stress with diseases, and the role of tryptophan metabolic pathway in the regulation of gut homeostasis, with particular focus on functional bowel disorders and their relationship to stress, and laid a theoretical foundation for the treatment of tryptophan in stress-related intestine diseases.
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To explore the protective effect of EGCG on acute kidney injury induced by cisplatin in rats based on Network Pharmacology

YANG CHUNXUE, XU ENSHUANG, ZHANG FENG, ZHENG JIASAN

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Objective To evaluate the protective effect of epigallocatechin gallate (EGCG) on acute kidney injury induced by cisplatin (CIS) in rats based on network pharmacology. Methods 32 male Wistar rats were randomly divided into four groups as follows: control group (CON group),EGCG group, CIS group (cisplatin group) and CIS+EGCG group. CON group and CIS group were given normal saline every day, EGCG and CIS+EGCG group were given EGCG (40 mg/kg) every day for 28 days. On the 26th day, CIS group and CIS+EGCG group were intraperitoneally injected with cisplatin (7 mg/kg).On the 29th day, blood and tissue were taken from each group. The levels of serum urea nitrogen (BUN) and creatinine (SCr) were detected, renal pathological changes were observed by HE staining, TUNEL was used to detect apoptosis in renal tissue. Drugs and disease targets were screened by TCMSP, Gene Cards and OMIM websites, protein-protein interaction network (PPI) was constructed after intersection, gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Gene and Genome (KEGG) enrichment analysis were carried out, and the results were verified by Western Blot 、RT-QPCR and immunohistochemistry. Results The results showed that EGCG preconditioning significantly decreased the levels of BUN and SCr in serum of AKI rats and improved the nephropathy of AKI rats, and relieved the renal tissue apoptosis of AKI rats. 87 EGCG and AKI intersection genes and 25 core targets were screened by network pharmacology, which affected the development of AKI through PI3K/AKT and other signal pathways and a variety of biological processes. Conclusions EGCG alleviates CIS-induced acute kidney injury in rats through PI3K-AKT signaling pathway.
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Study on establishment of mouse model of medial temporal lobe epilepsy with kainic acid

YE Zuliang, MIAO Yujing, LIU Quanlei, ZHU Quan, WEI Penghu

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Objective An ideal animal model of epilepsy can be used to study the pathogenesis of epilepsy and evaluate novel antiepileptic drugs. To optimize the simulation of the pathological characteristics and seizure behavior of medial temporal lobe epilepsy (MTLE), we aimed to establish a chronic epilepsy model of MTLE by unilateral, single hippocampal injection of kainic acid (KA) via stereotactic surgery, and to validate this epilepsy model in terms of behavior, electrophysiology, and pathology. Methods A total of 22 healthy C57BL/6 wild-type male mice were divided randomly into a control group (n = 6) and an experimental group, which received KA injections (n = 16). Both groups underwent microinjection of equal doses of saline or KA in the hippocampal CA3 area via stereotactic surgery. One week later, all mice were implanted with electrodes in the hippocampal CA3 area to facilitate electroencephalogram (EEG) recording. Seizure frequency and duration were analyzed statistically. The chronic epilepsy model was assessed in terms of behavior, electrophysiology, and pathology. Results No mice in the control group experienced seizures, while all surviving mice in the experimental group developed seizures. Adult model mice exhibited chronic spontaneous seizure behaviors, such as staring, chewing, head and facial muscle twitching, and limb spasms. Two mice died as a result of the surgery, four mice died during the acute seizure phase, and 10 model mice were successfully established. EEG recordings showed epileptiform changes consistent with MTLE. Immunofluorescence staining revealed neuronal loss in the CA3 area and astrocytic changes, consistent with characteristic pathological changes of hippocampal sclerosis. Conclusions We successfully established a chronic epilepsy model of MTLE by unilateral, single intracranial injection of KA and confirmed the validity of the model in terms of behavior, electrophysiology, and pathology. This KA model possesses several advantages, including being time-efficient, easy to perform, cost-effective in terms of drug usage, easily replicated, and being KA-dose-dependent. This epilepsy model exhibits similarities to human MTLE in terms of EEG, behavior, and neuropathological changes, and can thus be used to study effective drugs for treating temporal lobe epilepsy and as an ideal animal model for predicting the outcome of epilepsy surgery.
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Establishment and Optimization of Rapid Model of Osteoporosis in zebrafish

MAO Hongyun, LIU Yutong, ZHAO Xinyue, JIANG Deli, Wang Xiaoyi, ZHAO Kexuan, HUA Yongqing, Xu Huiqin

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Objective In this paper, a fast, stable and sensitive model of zebrafish osteoporosis (OP) was established by comparing different methods of establishing common zebrafish osteoporosis models. Methods The OP model was induced by iron overload and prednisolone (Pred). The bone formation and animal mortality were observed. Ammonium ferric citrate (FAC) was used as the model drug in iron overload induction method. The Pred induction method was Pred-3 dpf (day post fertilization) method, the modeling time was 3 ~ 9 dpf; Pred- 5 dpf method, the modeling time was 5 ~ 10 dpf; Pred withdrawal method, 3 dpf began to make the model, and 7 ~ 9 dpf was removed every day. Results The significant effect of FAC on bone formation was not observed at the dose of 500 μg/mL. Compared with the control group, the number of bone and the length of the first vertebra in the model group induced by Pred-3dpf decreased significantly (P < 0.01, P < 0.05), but the mortality of zebrafish was higher. There was no significant difference between Pred-5 dpf method (P > 0.05) Pred withdrawal method model group were significantly reduced (P < 0.01), and no death. AC, CA and AL all had anti-OP effect, and CA had more sensitive and stable anti-OP effect. The optimization results of dyeing parameters of alizarin red (ARS) showed that the dyeing concentration was 0.02%, and the washing conditions of 0.5% KOH and glycerol for 2 hours were the best. The results of chemical staining showed that calcein was more sensitive to the staining of bone nodes and ARS was more sensitive to the staining of the first vertebra. Conclusions The zebrafish OP model induced by Pred withdrawal method is more rapid, stable and sensitive, and it is a stable and reliable model for OP study.
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The impact of embryonic uveitis exposure on the offspring mice response to IRBP-induced experimental autoimmune uveitis (EAU)

Xufei, Liu Jianping, Dong Shifang, Huanghui, Gong xinyi, Hu kaijiao, Chen feilan

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Objective To investigate the effect of embryonic inflammatory exposure on the offspring response to IRBP-induced experimental autoimmune uveitis (EAU). Methods RNA transcriptome sequencing data from eyeball in the C57BL/6J offspring gestated from active parental EAU were used to screen immune-associated differentially expressed genes (DEGs) in the eyes of exposed offspring. Gene fragments overlapping in the two datasets were screened using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses to identify biological pathways associated with gene fragments. Hub genes were identified from these intersecting genes by protein-protein interaction networks (PPI) analysis. EAU models of offspring affected by maternal uveitis were established by immunization with IRBP651-670, and the expression levels of the pivotal genes in the offspring exposed to inflammation by maternal uveitis were examined by fluorescence quantitative polymerase chain reaction (q-PCR). The EAU severity, T lymphocytes proliferation and serum cytokines were detected to investigate the immune effect of offspring gestated from materinal active inflammation response to 150 μg IRBP induction. Results Microarray analysis identified 72 immune-related genes in ocular tissue DEGs compared with control samples. These genes were mainly enriched in pathways involved in Toll-like receptor signaling pathway, MAPK signaling pathway, and B cell receptor signaling pathway. Further PPI network interaction analysis screen out four hub genes, namely, Psmc5, Psmc3, Psmd4, and Psmd8. Meanwhile the adult offspring gestation from materinal active uveitis expressed increased mRNA level in those four hub genes compared with those healthy offspring. In addition, under 150 μg IRBP induction, an increase in the severity of EAU in the offspring during the inflammatory activity period was observed, with significant differences in clinical and pathological aspects compared with the control group. Meanwhile, the proliferation of T cells in the offspring during the inflammatory activity stage was enhanced, and the secretion of inflammatory cytokines IL-17 and IL-6 was increased. Conclusions Psmc5, Psmc3, Psmd4, and Psmd8 may be the important genes exacerbating offspring uveitis associated with the increased T cell proliferation and production of IL-17 and IL-6.
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Establishment of an orthotopic implantation model of hepatocellular carcinoma in mice

pan rui, yu kun, zhang hailiang, zheng yongren, zhao xiaoyu, tang junze, wu jianming, cheng xin

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【Abstract】Objective To explore the advantages and disadvantages of orthotopic injection of cell suspensions and transplantation of tumor tissue blocks to establish an orthotopic implantation model, and to provide technical reference for the establishment of an orthotopic implantation model. Methods A total of healthy Kunming mice were divided into four groups. Direct injection H22 cells was used in group A. Direct injection H22 ascitic cells was used in group B. Transplantation of tissues was used in group C. Direct injection was saline used in group D. The activity and weight change of each group were observed regularly. The survival time of the four groups of mice was recorded. Liver tumor formation, tumor size, abdominal organ adhesion degree and metastasis of the mice were observed. B-ultrasound imaging was performed to detect the concentration of alpha fetoprotein (AFP), abnormal prothrombin (DCP) and HE staining to observe liver histopathological changes. Results The mice in groups A, B and C were (3.36±0.44) min, (3.30±0.41) min and (5.68±0.65) min, respectively. After 25 d of modeling, the rate of mice in groups A, B and C was 100.0%, the severe abdominal adhesion in group A was 40.0%, 60.0% in group B, and neither group C or D. The rates of ascites in groups A, B and C were 40.0%, 100.0% and 0.0%, respectively; the rates of abdominal wall tumors were 30.0%, 60.0% and 0.0%, respectively. Mice in group B also had liver metastasis 40.0%). B ultrasound imaging, detection of serum AFP and DCP levels, histopathological results showed that mice in group A, B, C had smooth liver margins, uneven echo and slightly lower echo mass, maintaining AFP and DCP high secretion; large number of inflammatory cells and tumor cells. Conclusion H22 cells and H22 ascitic cells have the advantages of simple operation and high survival rate. transplantation of tissues has the advantages of less metastasis and light abdominal adhesion. All three methods can establish an orthotopic transplantation model of HCC, which can be an ideal model to evaluate the effect of drugs against HCC.
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Three Tiao-Bu Fei-Shen therapies can improve airway mucus hypersecretion in COPD rats by inhibiting ERK1/2 signaling pathway

LI Gaofeng, LIU Shujuan, LI Ya, LI Suyun, FAN Zhengyuan, SHEN Tingting

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Objective To investigate the three Tiao-Bu Fei-Shen therapies in improving airway mucus hypersecretion in stable chronic obstructive pulmonary disease (COPD) rats. Methods 90 rats were randomly divided into nine groups: control group, COPD group, Bu-Fei Jian-Pi (BJF) group, Bu-Fei Yi-Shen (BYF) group, Yi-Qi Zi-Shen (YZF) group, ERK1/2 inhibitor (PD98059) group, BJF + PD98059 group, BYF + PD98059 group and YZF + PD98059 group. The rat model of COPD was established by exposing them to cigarette smoke and repeated bacterial infection from the first to the eighth week. From the ninth to the sixteenth week, the Control group and the COPD group were given normal saline 2 ml, the TCM treatment group was given three Tiao-Bu Fei-Shen formulas by gavage, and the inhibitor group and the combination group were given PD98059 by intraperitoneal injection for 7 days. Lung function tests were conducted at the end of 16 weeks along with assessments on lung tissue morphology, lung water content,inflammatory cell count in bronchoalveolar lavage fluid (BALF), and level of inflammatory factors in serum. AB-PAS staining was used to determine goblet cell proportion while immunohistochemistry was employed to measure Muc5AC and Muc5B expression levels. PCR analysis was performed to detect ERK1/2 mRNA expression as well as ENaC, CFTR, AQP5 mRNA expressions. Western Blot analysis measured protein expression levels of ERK1/2and P-ERK1/2 in lung tissue. Results Compared with the Control Group, TV, MV, FVC, FEV0.1, FEV0.1/FVC significantly decreased (P<0.01) in COPD Group.Lung pathology revealed alveolar disorder, massive fracture of alveolar wall, and severe shrinkage/thickening of airway wall accompanied by extensive infiltration of inflammatory cells. Water content in lung tissue increased significantly (P<0.01). The proportion of macrophages in BALF was significantly reduced (P<0.01), whereas the proportions of neutrophils and lymphocytes were significantly increased (P<0.01). The level of TNF-α and IL-1β in serum were significantly increased (P<0.01). The percentage of goblet cells and expression levels of Muc5AC and Muc5B in airway epithelial cells exhibited a significant increase (P<0.01). Expression levels of ERK1, ERK2, and ENaC mRNA in lung tissue were significantly elevated (P<0.01), while expression levels of CFTR and AQP5 mRNA were significantly decreased (P<0.01). Compared to the COPD group, the treatment groups demonstrated improvements in the aforementioned indicators (P<0.05), with the three Tiao-Bu Fei-Shen formulas combined PD98059 group showing superior efficacy compared to single treatment groups (P<0.05). Conclusions Three Tiao-Bu Fei-Shen therapies can ameliorate airway mucus hypersecretion in COPD rats by inhibiting the ERK1/2 signaling pathway.
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Anti-inflammatory and analgesic effects of small intestine Ski-overexpressing on acetic acid-induced writhing mice

xiong ao, luo fei

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Objective: To study the anti-inflammatory and analgesic effects of Ski protein overexpression on mice induced by acetic acid.? Methods: Eight-week-old male ICR mice were selected and given 0.7% acetic acid solution (0.1ml/10g body weight) to induce writhing reaction.Experimental mice were divided into Sham group, acetic acid group, acetic acid + ibuprofen group, acetic acid +Ad-EGFP group, acetic acid +Ad-ski-1 group and acetic acid +Ad-ski-2 group, with 10 mice in each group. Observe and record the incubation period for the first appearance of twisting, while also recording the number of twisting times within 15 minutes. Obtain small intestine tissue, identify adenovirus transfection effect, detect protein expression levels of pro-inflammatory factors and pain biomarkers, and further analyze the protein expression of NF-kB p65 and its binding with Ski protein. Results:Ski protein was successfully overexpressed in small intestine after intraperitoneal injection of Ad-ski adenovirus. Overexpressed Ski protein prolonged the incubation period of writhing and decreased the frequency of writhing reaction, which was comparable to ibuprofen (P > 0.05). Compared with acetic acid group, overexpressed Ski protein significantly inhibited the protein expression of pro-inflammatory factors and pain biomarkers (*P < 0.05,**P < 0.01). Moreover, NF-kB p65 forms complexes with Ski. Conclusion: Overexpression of Ski protein has anti-inflammatory and analgesic effects on acetic acid induced inflammatory pain by inhibiting the expression of inflammatory factors and pain biomarkers, and is related to Ski regulation of the NF-kB signaling pathway.
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Exploration of the Establishment of an Experimental Animal Platform for Intelligent Acupuncture Robotic Acupuncture Feasibility and Safety Evaluation

MA Weigang, PAN Xingfang, QIU Jiwen, GAO Weifang, ZHANG Yonglong, DONG Yuge, TANG Yuzi, REN Haiyan, LI Zhongzheng

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Objective:The development of intelligent acupuncture robots is in the early exploratory stage and lacks a reliable experimental research platform. This study aims to explore the experimental animal platform for the acupuncture feasibility and safety evaluation of intelligent acupuncture robots, and to lay the foundation for further research.Methods: Six 2-month-old Guangxi Bama miniature pigs were used as experimental subjects for acupuncture verification after anaesthesia. Firstly, manual acupuncture verification was carried out, and 6 acupoints were selected for each experimental animal, and the needles were left for 20 min after the lifting, inserting and twisting manipulation, and before and after control was carried out. The experiment was carried out for 28 days, and the experiment was conducted once every two days for a total of 10 times. After the verification of manual acupuncture, a point was taken 10 mm beside each of the 6 selected acupoints, and a total of 12 points were taken, and the intelligent acupuncture module of the acupuncture robot was used to carry out acupuncture operations on the experimental animals at different frequencies and angles to further verify the stability and feasibility of the animal platform. Results: The safety-related indicators in the blood routine were not abnormal, and were stable compared with the before and after comparisons of themselves. Blood biochemistry indicators were not abnormal, and were stable compared with the before and after comparisons. The average heart rate of the animals was 126 beats/min, and the average blood pressure was 87/36 mmHg. The average blood pressure of the animals was 87/36 mmHg, and the average body temperature of Bama miniture pigs was 36℃ at room temperature of 25℃. There was no significant change in body temperature during and after the experiment. On the basis of the experimental platform, the pre-test of acupuncture manipulation with the intelligent acupuncture module of the acupuncture robot was successfully completed, and there were no abnormalities related to acupuncture such as bending, breaking, or stagnation of needles during the experimental process, and the experimental animals did not show any obvious abnormality.Conclusion:In this study, a stable experimental animal platform for the evaluation of acupuncture feasibility and safety of intelligent acupuncture robots was initially established by drawing on the existing experimental method of miniature pigs, laying a foundation for further research related to intelligent acupuncture robots to be carried out.
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The effect and mechanism of olaparib on the senescence of MCF-7 breast cancer cells

Wang Dawei, Guo Jing, Bian Jichun, Wang Shasha, Lu Meichao, Zhang Daizhou, Jia Yuping

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Objective To study the performance and related molecular mechanism of Olaparib-induced senescence in MCF-7 breast cancer cells. Methods Real-time cell analysis (RTCA) was used to dynamically detect anti-proliferation and anti-migration activities in real time. Senescence-associated β-galactosidase (SA-β-gal) staining was used to observe the senescence-inducing activity of cells. The effects of olaparib on the expression of senescence-associated genes p16INK4a, p21, C/EBP homologous protein (CHOP) , interleukin (IL) -6, IL-8, plasminogen activator inhibitor 1 (PAI-1) , phosphatase and tensin homolog deleted on chromosome 10 (PTEN) , p27, retinoblastoma protein (RB1) , Ki67 and E2F1 were analyzed by qPCR. The effects of olaparib on the expression of senescence-associated proteins p21, γH2AX, pRB, cyclin D1, insulin-like growth factor binding protein 3 (IGFBP3) and Ki67 were analyzed by Western blot. Results Olaparib could inhibit the proliferation and migration of MCF-7 breast cancer cells and induce the senescence of MCF-7 cells. The gene expression levels of p16INK4a, p21, p27, CHOP, IL-6, IL-8, PAI-1, PTEN and RB1 in MCF-7 cells treated with olaparib for 96 h were significantly up-regulated (P < 0.01) , and the gene expression levels of Ki67 and E2F1 were significantly down-regulated (P < 0.01) . The expression levels of p21, γH2AX and IGFBP3 proteins in MCF-7 cells were significantly increased (P < 0.01, P < 0.01, P < 0.05) , and the expression levels of cyclin D1, pRB and Ki67 proteins were significantly decreased (P < 0.05, P < 0.01, P < 0.05) . Conclusion Olaparib can produce anti-MCF-7 breast cancer cells by anti-proliferation, migration and induction of cell senescence.
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The Protective Effect and Mechanism of BBD on Doxorubicin-induced Myocardial Injury in Zebrafish model

LI Xianmei, ZENG Laifeng, HUANG Bin, YU Lanxin, LIN Jiumao

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Objective To investigate the protective effect and mechanism of BBD on Doxorubicin (Dox) induced myocardial injury in zebrafish. Method A zebrafish model of myocardial injury induced by chemotherapy drug Dox was established. The effects of BBD at different concentrations on pericardial edema ratio and heart rate were observed under a stereo microscope. Transgenic zebrafish Tg(mpx:EGFP) was used to observe the inhibitory effect of BBD on neutrophil infiltration in the heart in situ., and observe the effect of BBD on SOD, CAT and MDA. Real-time qPCR was used to detect the expression of ferroptosis related factors gpx4a, ptgs2, alox5a, acsl4. The accumulation of ferrous ion in zebrafish heart was detected using a ferrous ion fluorescent probe. Results BBD significantly improved Dox induced pericardial edema, heart rate and neutrophil infiltration in zebrafish (P<0.05), significantly increased the reduction of SOD and CAT activity (P<0.05), and decreased the concentration of MDA (P<0.05). Compared with the Dox group, the Dox group showed significantly increased expression of gpx4a (P<0.05) and decreased expression of ptgs2, alox5a, and acsl4 (P<0.05), and significantly inhibited Dox induced ferroptosis in zebrafish hearts. Conclusion BBD can attenuate Dox induced myocardial injury in zebrafish by inhibiting the occurrence of lipid peroxidation and regulating ferroptosis.
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The metabolic profiles of serum lysophosphatidylcholine and amino acid in rats with non-alcoholic fatty liver disease

ZHAO Mei-yu, SHI Xin-yue, ZHOU Shu-ling, LI Hai-jun, FAN Shu-ting, XIONG Yin-hua

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Objective: Using metabolomics methods to study the metabolic profiles of amino acids and lysophosphatidylcholine (LPC) in serum of rats with non-alcoholic fatty liver disease(NAFLD), and to identify biomarkers that can characterize NAFLD and speculate on the possible mechanism of its occurrence. Methods: NAFLD rats were prepared by feeding high-fat diet feed and intraperitoneal injection of carbon tetrachloride. The levels of 15 LPCs and 18 amino acids in the serum of the control and NAFLD groups were determined using liquid chromatography-mass spectrometry.SPrincipal component analysis and orthogonal partial least squares discriminant analysis were used to analyze the changes in serum LPC and amino acid metabolic profiles in NAFLD rats. Pearson correlation analysis was used to analyze the correlation between biomarkers and NAFLD.SResults: The metabolic profiles of serum LPC and amino acid in the NAFLD group significantly deviated from the control group and could be completely distinguished.SLPC (20:1), arginine and glutamic acid had significant contributions to NAFLD, and these species were identified as biomarkers. Furthermore, LPC (20:1) and arginine were significantly correlated with serum biochemical indicators such as AST, ALT, LDL, and TBIL.SConclusions: The metabolic profiles of serum LPC and amino acid were closely related to NALFD.
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The creation and translational relevance of abdominal aortic aneurysm animal models

Xia Congcong, Liu Haole, Hou Haiwen, Liu Enqi, zhao sihai

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With the population aging in our country, the incidence of abdominal aortic aneurysm(AAA), an aged disease, has gradually increased. Because of the high mortality of ruptured AAA, which became one of the most severe life-threaten diseases. In histopathology, main AAA pathological features include elastin degradation, smooth muscle cells (SMCs) depletion, extracellular matrix digestion, and mural leukocytes accumulation. In clinics, open/endovascular repair remains the effective strategy for AAA management, and the drug therapy is still lacking. The detailed molecular mechanism of AAA is still unclear, which is one of the important bottlenecks affecting the development of potential drug targets. Animal models are the most powerful tools to clarify the pathogenesis of AAA. Though some medium to large laboratory animal models (such as rabbits, guinea pigs, dogs and pigs) are established for AAA study, rodent models (mice and rats) still take the first place in this field. At present, the methods to induce AAA include intra-infrarenal aortic infusion of elastase, subcutaneous infusion of angiotensin II (Ang II), periaortic calcium chloride painting, decellularized aortic xenograft and so on. For the cease of pre-induced stimulation (medical or surgical), AAA tends to stabilize in most models. Therefore, the development of ideal animal models, which maintains the continuous aortic dilation and even rupture, remains a problem. AAA animal models are helpful in elucidating the pathogenesis, screening new drug targets and promoting its clinical translation. This review aims to discuss the application of current AAA modeling methods and their translational relevance.
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Establishment of Liver-specific RBP4 Transgenic Mice and Analysis of Glucose Metabolism Characteristics

LU Wanxian, MA Qi, WANG Li, LIU Mengdi, GUO Baoping

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Objective To construct a liver-specific Rbp4 knockout mouse model and to explore the effect of liver RBP4 on glucose metabolism. Methods Homozygous liver-specific Rbp4 knockout (Rbp4flox/flox:Cre+) mouse model was obtained by multiple breeding hybridization between LoxP-labeled Rbp4flox/+ mice and Alb-Cre tool mice. The genotype of mice was identified by agarose gel electrophoresis. Eighteen-week-old male mice were selected and divided into liver-specific Rbp4 knockout mice group (Rbp4flox/flox:Cre+, n=10), littermate control mice group ( Rbp4flox/flox, n=10) and wild-type mice group (WT, n=10). Western blot and qRT-PCR were used to detect the expression levels of Rbp4 protein and mRNA in liver and the expression levels of Rbp4 mRNA in other tissues to verify the specific knockout results. HE staining was used to observe the morphological changes of liver tissue in mice. Glucose tolerance and insulin tolerance were measured at the end of the 18 th week. The expression of liver glucose metabolism genes phosphoenolpyruvate carboxylase ( Pepck ) and glucose-6-phosphatase ( G6pase ) was detected by qRT-PCR. Results Western blot and qRT-PCR results showed that the expression of Rbp4 in the liver of Rbp4flox/flox:Cre+ group was decreased (P< 0.05 ). HE staining, glucose tolerance and insulin tolerance results showed that liver-specific Rbp4 knockout had no significant effect on liver morphology, glucose tolerance and insulin tolerance. Compared with WT group, the relative expression of Pepck mRNA in Rbp4flox/flox group was increased, and the difference was statistically significant (P＜0.05). Compared with WT group, there was no significant difference in the relative expression of Pepck mRNA in the liver of Rbp4flox/flox:Cre+ group (P> 0.05 ).Compared with the Rbp4flox/flox group, the relative expression of Pepck mRNA in the liver of the Rbp4flox/flox:Cre+ group was decreased, and the difference was statistically significant (P＜0.05). There was no significant difference in the expression of G6pase mRNA in the liver of the three groups (P> 0.05 ). Conclusion A liver-specific Rbp4 knockout mouse model was successfully constructed. Under physiological conditions, liver-specific Rbp4 knockout had no significant effect on liver tissue morphology, glucose tolerance and insulin tolerance in mice, and reduced the expression of liver glucose metabolism gene Pepck, suggesting that RBP4 plays a role in liver glucose metabolism.
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Progress in the study of animal models of hemophilic arthritis

LIN KUN

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Hemophilia Arthritis (HA) is caused by recurrent bleeding, which seriously affects the life of patients and consumes a lot of social and medical resources. Limited by ethical requirements, it is necessary to establish an animal model of HA for research. In order to summarize the research progress of animal models of hemophiliac arthritis, this paper reviewed the literature on animal HA models at home and abroad in recent years, from the aspects of species selection, modeling methods, histopathology, imaging evaluation methods, etc.
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A review of studies on the construction of experimental animal models and evaluation methods of spleen deficiency syndrome

ZHANG Yonglong, MA Weigang, QIAN Xingyu, ZHAO Suhong, LI Shanshan, GUO Yi, GUO Yongming, XU Zhifang, PAN Xingfang, QIU Jiwen, LI Zhongzheng

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The construction of experimental animal models of Chinese medicine syndromes plays an important platform support role in the research of the mechanism of action of Chinese medicine. In recent years, there are more and more reports in the literature on the construction and evaluation of the animal model of spleen deficiency evidence, but there are different standards of the construction method and insufficient objectification of the evaluation indexes. In this paper, we will summarize the construction and evaluation methods of the animal model of spleen deficiency from the aspects of animal selection, model establishment, macroscopic characterization, behavioral experiments, and objective indexes of spleen deficiency. evaluation methods, with a view to providing theoretical guidance for the construction of experimental animal models of spleen deficiency and literature reference for the selection of animal model platforms for spleen deficiency.
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Research Progress in Mouse Model of Sarcopenia

Jia Xinao, Tang Yujiao

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Through the study of literature retrieval related to sarcopenia, the modeling method and model evaluation scheme of sarcopenia mice were summarized. The operation methods, advantages and disadvantages, and application scope of the four modeling methods of drug injection, aging, muscle atrophy, and transgenic were reviewed. The evaluation methods of muscle function, muscle strength, and muscle endurance were summarized, and their advantages and disadvantages were compared to provide reference for the later research of sarcopenia.
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Therapeutic effects of Isaria feline combined with cyclophosphamide in hepatoma H22 tumor-bearing mice

SHI Xiaowei, CHEN Jingjing, YU Guoyan, ZHANG Yiyin, CHEN Lixia, ZHAO Lili, YANG Yongming, WANG Jing, YAN Lei, YANG Xihua

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【Abstract】Objective To investigate the therapeutic effects of Isaria felina originated from cordyceps sinensis combined with cyclophosphamide in hepatoma H22 tumor-bearing mice. Methods The model of H22 tumor-bearing mice was established and twenty-five Kunming mice were randomly divided into one blank control（NC）and four H22 tumor-bearing groups including tumor model control（MC）, positive control（CTX）, Isaria felina group（IF）and combined administration group（IF + CTX）. The mice in the blank control group and the model control group received distilled water by oral gavage, the mice in the positive control group were intra-abdominally injected with CTX olution（25 mg/kg body weight every day）, the mice in the IF group were orally administered Isaria felina（dispersed in distilled water） at 400 mg/kg body weight, and mice in the combined administration group were administerd the above two medications per day. The adminisation cycle was 10 d. At the end of the experiment, the mean tumor volume and weight, tumor inhibition rate, q value and immune organ index were calculated, and the levels of blood routine and cytokines were determined. The pathomorphological changes of tumor tissues were observed by HE staining. Results the tumor volume and mass of mice in each treatment group were significantly lower than those in MC group（P ＜ 0.05）. The tumor inhibition rates of CTX, IF and IF + CTX groups were 49.3%, 34.2% and 72.8%, respectively and the q value was 1.09. The number of WBC, Lymph and PLT in IF + CTX group was significantly higher than that in CTX group（P ＜ 0.05）. The spleen index of MC group was significantly higher than that of NC group, and the spleen index of IF + CTX group was significantly lower than that of MC group（P ＜ 0.05）. The level of serum IFN- γ in MC group was significantly lower than that in NC group, while the levels of IFN- γ in IF and IF + CTX groups were significantly higher than that in MC group and CTX group（P ＜ 0.05）. Pathologically，the tumor cells in the MC group grew well, numerous and closely arranged. While in the CTX, IF and IF + CTX groups, tumor cells arranged loosely, focal necrosis and nuclear pyknosis of necrotic cells could be seen in many places. Conclusion The combination of IF and CTX has an additive anti-tumor effect on H22 tumor-bearing mice, which can reduce immunosuppression and myelosup-pression caused by CTX, and have immunomodulatory function. 【Key words】 Isaria felina; H22 tumor-bearing mice; combined administration; immunomodulation
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Establishment of qPCR method for detection of Staphylococcus xylosus and its application

YU LINGZHI, FENG LIPING, KONG ZHIHAO, ZHU QI, WEI XIAOFENG

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Objective To establish and evaluate a method for rapid and sensitive Staphylococcus xylosus detection using qPCR. Methods This study selected the specific gehM gene fragment as the target for Staphylococcus xylosus, synthesized a set of specific primers and established a qPCR method for detection of Staphylococcus xylosus. Staphylococcus xylosus standard strain and other non taeget strains were chosen for specific analysis. Diluted the DNA of Staphylococcus xylosus to 10 times to determine its sensitivity. Clinical samples were tested and positive products were sequenced. The results were compared with those of bacterial culture. Results Staphylococcus xylosus had a specific amplification curve, while other non-Staphylococcus xylosus species did not show it, indicating that the primers were specific for Staphylococcus xylosus. The sensitivity was 100 fg DNA. The repeatabilities within and between groups are less than 3%. A total of 60 clinical samples were detected, of which 5 samples had a typical S curve. The PCR products were sequenced and BLAST. The similarity of the gene sequence was 99.63%, indicating that the sample was positive for the nucleic acid of Staphylococcus xylosus gehM gene, with a positive rate of 8.3%. However, the positive rate of bacterial culture was 6.7%. The positive rate of qPCR was slighterly higher than the culture. Conclusions The qPCR method established has the advantages of fast, high sensitivity and specificity, and can be used for the detection of Staphylococcus xylosus.
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Guanxinning tablet improves doxorubicin-induced cardiomyocyte pyroptosis in rats with heart failure by inhibiting NLRP3/ASC/Caspase-1 pathway

shi jia jun, yang qin qin, fu dan ting, zheng chun wei, zhang yan, chen yu

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【摘要】目的研究冠心宁片对扩张型心肌病（dilated ca rdiomyopathy，DCM）的保护作用，并从NLRP3/ASC/Caspase-1通路深入探讨其对心肌细胞焦亡的作用机制。方法随机将大鼠分为冠心宁片低剂量组、冠心宁片高剂量组、阳性药地高辛组、模型对照组和正常对照组，通过阿霉素（doxorubicin，DOX）累积注射17.5 mg/kg诱导大鼠DCM模型，同时连续给药10周。超声心动图检测心功能指标，ELISA检测血清IL-1β和IL-18水平，RT-PCR法检测心肌组织NLRP3、ASC、Caspase-1、NF-κB、TXNIP、IL-1β和IL-18的mRNA表达，免疫组化、免疫荧光染色和western blotting检测NLRP3、ASC、Caspase-1、IL-1β、IL-18、GSDMD、GSDMD-NT的蛋白表达及TUNEL染色结果，透射电镜观察心肌细胞显微结构的变化。结果与正常对照组比，模型对照组的IVSs、IVSd、LVPWs、FS、SV、EF和HR显著降低，LVIDs、ESV及血清IL-1β、IL-18显著增加，NLRP3、ASC、Caspase-1、NF-κB、TXNIP、IL-1β和IL-18的mRNA表达均显著增加，NLRP3、ASC、Caspase-1、IL-1β、IL-18的蛋白表达及TUNEL染色面积明显增加，显微结构明显改变。与模型对照组相比，冠心宁片可显著增加IVSs、SV、FS、EF和HR，显著降低LVIDs、ESV和血清IL-1β、IL-18水平，降低心衰大鼠的NLRP3、ASC、Caspase-1、NF-κB、TXNIP、IL-1β、IL-18的mRNA和NLRP3、ASC、Caspase-1、IL-1β、IL-18的蛋白水平及TUNEL染色面积，显微结构明显改善。结论冠心宁片可以有效改善阿霉素诱导的扩张型心肌病，可能是通过抑制NLRP3/ASC/Caspase-1通路改善扩张型心肌病大鼠的心肌细胞焦亡实现的。
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Preparation of a rat model of diarrheal irritable bowel syndrome induced by acetic acid enema combined with binding tail-clamping stress

laibiyu, hongmengying, heyongjia, chenyao, lixinwu, lixing, she chang, lidan, shijia, tianzihan

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【Abstract】 Objective To establish an ideal modeling method for Diarrhea Predominant Irritable Bowel Syndrome (IBS-D) with anxiety and depression in rats, and to provide an animal model for clinical study of IBS-D (IBS-D). Methods In the first stage, 20 rats were randomly divided into blank group, 3% acetic acid enema group, 4% acetic acid enema group and 5% acetic acid enema group. On the first day of intervention, the rats were given acetic acid enema with 3%, 4% and 5% concentration, respectively. After observation for 1 week, the appropriate acetic acid concentration was evaluated. In stage 2, 40 rats were randomly divided into blank group, acetic acid enema group, acetic acid + binding group, acetic acid + binding + tail clip group, and were treated with different interventions for a total of 8 days. After the intervention, the general condition, diarrhea-related index, open field test score and colonic histopathology of rats were evaluated. Results In the first stage, compared with the blank group, the fecal trait score of 4% acetic acid enema group increased at 1-3 days after intervention (P < 0.05), and gradually decreased at 4-7 days after intervention, and there was no significant difference between the group and the blank group at 7 days (P > 0.05), and the body mass was relatively lower at 1 week after intervention (P < 0.05). Fecal water content increased (P < 0.01); Compared with blank group, body mass of 5% acetic acid enema group was decreased (P < 0.05), fecal trait score, fecal water content and diarrhea index were increased (P < 0.01). There was no significant difference between 3% acetic acid enema group and blank group. Compared with the blank group in the second stage, the body mass of the simple acetic acid enema group was lower (P < 0.01), and the fecal water content and diarrhea index were increased (P < 0.01). Compared with the blank group and the simple acetic acid enema group, the body mass of the acetic acid + restraint group and the acetic acid + restraint + tail clip-on group was lower (P < 0.01). Fecal trait score, fecal water content and diarrhea index increased (P < 0.01), and colon running time decreased (P < 0.01). In the open field test score, compared with the blank group and the simple acetic acid group, the open field test distance, standing times and upright times in the acetic acid + bound group and the acetic acid + bound + clip group were all the same (P < 0.01). Compared with acetic acid + binding group, the open field experiment distance, standing times and upright times in acetic acid + binding + tail group were all different (P < 0.05). The pathological tissue of colon showed that compared with the blank group, the mucosa structure of 4% acetic acid enema group was complete with a small amount of inflammatory cell infiltration, and the pathological tissue score had no significant difference (P > 0.05), while the 5% acetic acid enema group had a medium to large amount of inflammatory cell infiltration, and the pathological tissue score was increased (P < 0.05). The mucosal structure of colon tissues in each group in stage 2 was intact, and a small amount of inflammatory cells infiltrated in different degrees. Conclusion 4% acetic acid concentration is the appropriate concentration for IBS-D modeling. After superposition and binding, IBS-D diarrhea and internal hypersensitivity characteristic state can be better simulated. After superposition of tail clip, IBS-D model of liver-stagnation and spleen-deficiency can be successfully prepared.
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The protective effection on allergic asthma rats of Xiebaisan based on Non-targeted Metabolomics and Intestinal bacterial flora

yangzongtong, Xu Dongchuan, Liu Jin, LI Xiaojing, Zhang Huimin, WANG Wenhui, Sui Zaiyun

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【Abstract】Objective To explore the possible mechanism of Xiebaisan in protecting allergic asthma rats from the perspective of host-intestinal flora-metabolism. Methods The SPF SD rats were divided into normal group (NC Group) , model group (M group) and Xiebaisan Group (Xiebaisan Group); Allergic asthma rat model was established by ovalbumin (OVA); The changes of lung histopathology in rats were observed by stained with Hematoxylin-HE. Colon contents were harvested for 16S rRNA high-throughput sequencing to observe the changes of intestinal flora structure and function; Serum and lung tissue samples were collected for non-targeted metabolomics analysis by UHPLC-Q-TOF/MS. Results The results of HE showed that, Xiebaisan group could improve the lung histomorphology of asthmatic rats to some extent compared with M Group; The results of 16S rRNA high-throughput sequencing showed that, the diversity of intestinal flora decreased in M group, and the diversity of intestinal flora was improved in Xiebaisan group compared with M group; The results of serum non-targeted metabolomics test showed that, Xiebaisan group could regulate amino acid metabolism and mTOR pathway, and reversed the differential metabolite expression induced by M group partially; The results of non-targeted metabonomics of lung tissue samples showed that, the Xiebaisan group could regulate carbon metabolism, vascular smooth muscle and cAMP signaling pathways, and reversed the differential metabolite expression induced by M group partially. Conclusion The protective effects of Xiebaisan on allergic asthma in rats may be related to the improvement of the morphological structure of lung tissue, the diversity of intestinal flora, and the regulation of mTOR pathway, Vascular smooth muscle contraction pathway and cAMP pathway, which affect the metabolism of amino acids and carbon.
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Construction and Evaluation of a Mouse Model of Chronic Restraint Intestinal Stress Injury

ZHENG Jianhua, CHEN Jingqing, DONG qiaoyan, FA Yunzhi, QIU Yefeng

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Objective Given that psychosocial stress can contribute to a series of diseases, such as inflammatory bowel disease and irritable bowel syndrome, we aimed to establish an experimental chronic restraint mouse intestinal stress injury model as a basis for exploring the pathogenic mechanism of chronic restraint stress-induced gastrointestinal diseases, and for developing preventive and curative measures. Methods Eighteen male SPF-grade BALB/c mice were acclimatized for 7 days and then divided into a control group and a chronic restraint stress group according to body weight, using a randomized numerical table method. The mice were subjected to restraint stress for 3 hours per day for 14 days to establish an intestinal injury model. The model was evaluated by observing body weight, pathological changes in intestinal histomorphology, expression of tight junction proteins, apoptosis of intestinal epithelial cells, and mRNA expression levels of inflammatory cytokines. Results After 14 days of chronic restraint stress, model mice showed weight loss, shortened duodenal villus height, abnormal crypt structure, a decreased villus/crypt ratio, colonic mucosal inflammatory cell infiltration, and irregular crypt structure. Protein immunoblotting, immunohistochemistry, and immunofluorescence staining showed that the expression levels of the duodenal and colonic tight junction proteins occludin and claudin-1 were significantly decreased in mice after chronic restraint stress (P < 0.05), while expression levels of the apoptotic protein cleaved-caspase-3 in intestinal epithelial cells were significantly increased (P < 0.05). Regarding the mRNA expression levels of intestinal inflammatory factors and chemokines, chronic restraint stress for 14 days significantly increased the gene expression levels of interleukin (IL)-1β, IL-6, monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α, and IL-10 in the duodenum of mice (P < 0.05), and significantly increased the gene expression levels of IL-1β, IL-6, and MCP-1 in the colon (P < 0.001). Conclusions The use of a behavioral restriction device to restrain mice continuously for 14 days led to abnormal intestinal tissue structure, intestinal barrier dysfunction, and intestinal epithelial cell apoptosis, and triggered an intestinal inflammatory response in the stressed mice, indicating successful establishment of a mouse model of intestinal injury by chronic restraint stress.
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Establishment and Analysis of NLRP3-/- Mouse Models of Ulcerative Colitis

wangzhuhuan, zhangerxin, zhengqinwei, haoweiwei

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To induce NLRP3-/- mouse ulcerative colitis (UC) model using different concentrations of Dextran Sulfate Sodium (DSS) and administration time, and to analyze and evaluate the advantages and disadvantages of its preparation, in order to provide a more clinical animal model for the study of UC pathogenesis and the development of therapeutic drugs.Method: 48 NLRP3-/- mice of SPF level were randomly divided into groups and induced UC mouse models using a combination of different concentrations of DSS and administration time (blank group, 2.5% 7 days, 3% 7 days, and 3% 5 days). The physical weight, DAI score, HE staining, measurement of colon length and related indicator molecules (IL-6, TNF) of the mice were observed and evaluated- α Compare the effect of modeling with the expression level of tight junction protein.Result: 1. Each group was able to induce UC membrane type at different concentrations and administration times; 2. As the concentration gradient increases and the administration time prolongs, the physical weight of the mice decreases more significantly, the fecal occult blood is more positive, the DAI score is higher, and the mice die; 3. HE staining showed that the longer the time and the higher the concentration of DSS, the more severe the damage to the intestinal mucosal tissue in mice; 4. Using immunohistochemistry to detect inflammatory factors and tight junction proteins, compared to the blank group, the inflammatory factors (TNF-α) in the model group were detected，The expression levels of IL-6 and IL-6 increased, while the expression level of tight junction protein (ZO-1) decreased compared to the blank group.Conclusion: 1. NLRP3-/- mice can induce UC models under conditions of 2.5% 7 days, 3% 7 days, and 3% 5 days; 2. Combining DAI score, HE staining, and related indicators detection, as well as mouse survival rate, NLRP3-/- mice induced UC model under 3% DSS-5d conditions are more suitable for clinical manifestations of UC and more favorable for later drug intervention.
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The AOM/DSS Inflammation-cancer Transformation Model Optimized and Explored the Changes of Intestinal Microbiota in Mice with Colon Cancer

ZHU Lin, WANG Dunfang, FENG Xue, ZHANG Caijuan, LIU Haifan, LIU Yaqing, LIU Bin, LIU Li, YANG Weipeng

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【Abstract】 Objective To optimize the method of combining azomethane oxide (AOM) and dextran sodium sulfate (DSS) to create a colitis-associated colon cancer (CAC) model, and to explore the pathogenesis of the intestinal flora in CAC. Method Model groups A and B were established by one and two injections of AOM, respectively, combined with free drinking of DSS, and a normal control group was injected intraperitoneally with normal saline combined with purified water (n=10 mice per group). The better modeling scheme was selected by comprehensive evaluation of the disease activity index score, colon length, tumor rate, and mortality. Serum levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and tumor markers CA199, CEA, and CA724 were detected by enzyme-linked immunosorbent assay. Colon lesions were evaluated by hematoxylin and eosin (HE) staining. Changes in the intestinal microbiota in CAC mice were detected by 16S rDNA high-throughput gene sequencing analysis of mouse feces. Results Both single and enhanced AOM injections combined with DSS induced CAC mice; however, colon growths were larger, more closely arranged, and their morphological size was more consistent in group B compared with group A, with a tumor-formation rate of 100%. IL-6 and TNF-α levels were increased in the model group compared with the normal group (P<0.05, P>0.05, respectively). The CA199 and CEA levels were also significantly increased (P<0.05), but CA724 levels were not. Infiltration of inflammatory cells in the colon detected by HE pathology was accompanied by high-grade intraepithelial tumor-like changes on the surface of the lumen. The diversity and abundance of intestinal bacteria were decreased in CAC mice compared with normal mice: phyla Verrucomicrobiota and Actinobacteriota were significantly increased (P<0.05), Bacteroidota and Campilobacterota were significantly decreased (P<0.05). Akkermansia, Prevotellaceae, Ruminococcus, and Bifidobacterium were significantly increased (P<0.05), and Roseburia, Rikenellaceae_RC9_gut_group, Anaeroplasma, and Muribaculaceae were significantly decreased (P<0.05). Conclusion Two injections of AOM combined with 1.5% (1.5 g/100 ml) DSS induced CAC model mice with a high colon-tumorigenesis rate, uniform tumor morphology, and low mortality, and may thus be the preferred modeling scheme for pharmacodynamic experiments. Disorders or dysfunction of the intestinal flora may lead to increased permeability, loss of intestinal mucosal barrier function, and the release of enterogenic endotoxins, resulting in a sustained inflammatory response, as an indirect or direct cause of CAC pathogenesis.
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Construction of HIF-1α gene knockout plasmid and functional verification of naked mole rats based on CRISPR/Cas9 system

Zhang Jingyuan, JIANG Xiaolong, CUI Shufang

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Objective Plasmids construction and knocking out HIF-1α gene of NSF cell line by CRISPR/Cas9 genomic editing technology, providing an in vitro cell model for studying the mechanism of hypoxia tolerance and the occurrence and development of hypoxia related diseases in naked mole-rats. Methods Design four pairs of sgRNA sequences which targeted exon 1-4 of NSF HIF-1α gene and successfully construct an expression plasmid. The plasmid with optimal sgRNA was identified and transfected into 293T cells, and the supernatant was used for detecting the virus titer. Furthermore, lentivirus particles carring sgRNAs of HIF-1α infected NSF cells which have been infected with Cas9 lentivirus particles based on the protocol of manufacture previously. After drug screening of post transfection, fluorescence signals were observed under fluorescent microscope, and the expression of HIF-1α in NSF cells were detected by Western blot and T7E1 enzyme. Results The Sanger sequencing results showed that the designed sgRNA was successfully inserted into pX459 and pKLV2-U6-sgRNA2 vectors, demonstrating that the recombinant plasmid used for the transfection were successfully constructed;The T7E1 digestion experiment successfully removed 3 bands, the target efficiency of sgRNA was 54%, and the Western blotting results showed that the HIF-1α gene was successfully knocked out and the protein level was significantly reduced in NSF cells of naked mole rats (P=0.0019). Moreover, no obvious changes in the morphology of HIF-1α knockout cells were found under the microscope, and gene knockout had no obvious effect on cell proliferation. Conclusion The HIF-1α knockout cell line was successfully constructed using CRISPR/Cas9 technology, which will provide experimental basis for the further study of the HIF-1α biological function. Furthermore, it will be beneficial for the study of the mechanism of hypoxia tolerance in naked mole mice and provide theoretical foundation for the prevention and treatment of hypoxia related diseases.
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Development of inflammation and coagulation disorders in Sepsis

hou yuan lu, zhao ru ru, gao lei, li qi feng, yao zheng, li ming hong

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Objective:To study the changes in coagulation function and inflammation levels during sepsis. Methods: Multiple infection sepsis model (MIM), based on cecal ligation and puncture (CLP), was apllied to establish a rat model of sepsis. Forty-eight male SD rats were randomly divided into blank group (control group, n=8), sham operation group (sham group, n=8), 4-hour sepsis group (4h group, n=8), 8-hour sepsis group (8h group, n=8), 12-hour sepsis group (12h group, n=8), and 16-hour sepsis group (16h group, n = 8). ELISA and coagulation analysis were used to detect inflammatory markers and coagulation-related indicators. Results: (1) Compared with the sham group, the content of LPS and IL-6 in the model rats of all time points increased significantly(p<0.001). The LPS and IL-6 gradually increased as the disease progressed, however no significant changes after 12 hours. (2) In the middle and late stages of the septic model(starting from the 8h), PT was significantly prolonged compared with that in the sham groups(p<0.01). (3) Compared with the sham group, APTT time was significantly prolonged in 8h,12h,and 16h groups(P<0.05,P<0.01). The APTT time gradually lengthened from 8h group, and gradually approached control group after that. (4) Except for 8h group, the Fbg content in septic groups of all time points increased significantly than sham group (p < 0.01). (5) There was a significant difference between control group and sham group in FDP (p < 0.01), but no significant difference between sham group and septic groups. (6) Compared with the sham group, the level of AT-III in each period of progression in sepsis was decrease(p < 0.01), and there were significant differences in the proportions of 4h group,8h group, and 16h group. Conclusion: In the progression of sepsis, infection and inflammation levels gradually increase and cause coagulation dysfunction. The Fbg increases rapidly in the disease progresses, indicating the rapid activation of ogenous after model surgery. However, the PT and APTT time significantly increase in the middle and late stages of sepsis, suggesting the coagulation factors gradually depleted to induce DIC. At the same periods, multiorgan damage caused by sepsis leads to reduced expression of AT-III, further aggravating the coagulation/anticoagulant disorders. In addition, during the disease progression, a large amount of FDP interferes with fibrin polymerization, resulting in hemorrhagetendency and worsening DIC.
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WNK2 delays the proliferation and invasion of HCC by inhibiting tyrosine phosphatase activity

wuxueliang

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Abstract: Objective To investigate the effect of WNK2 on the expression of SHP2 (tyrosine phosphatase) in HCC and to explore the effect on the proliferation and migration of HCC. Methods HepG2 cells were transfected with WNK2-mimic, WNK2-inhibitor, and corresponding negative control. The effect of WNK2 on the proliferation of HCC was examined by subcutaneous tumorigenesis assay in Balb/c nude mice. The expressions of WNK2, p-SHP2, p-AKT, and p-ERK1/2 in tumor tissues were detected by Western blot. After treatment with SHP2 inhibitor PHPS1, the expressions of WNK2, p-SHP2, p-AKT, and p-ERK1/2 in HepG2 cells were detected by Western blot. The migration ability and invasion ability of HepG2 cells were detected by cell scratch assay and Transwell. The proliferation ability of HepG2 cells was detected by monoclonal proliferation assay. Results Compared with the NC group, the tumor volume of nude mice in the sh-RNA WNK2 group was significantly increased (p<0.01), while that of nude mice in the WNK2-mimic group was significantly decreased (p<0.01). Western blot results showed that compared with the NC group, the expression of WNK2 in the sh-RNA WNK2 group was significantly decreased (p<0.01), while the expressions of p-SHP2, p-AKT, and p-ERK1/2 were significantly increased (p<0.01). However, the expression of WNK2 was significantly increased in the WNK2-mimic group (p<0.01), and the expressions of p-SHP2, p-AKT, and p-ERK1/2 were significantly decreased (p<0.01). In vitro experiment, compared with the NC group, the expression of WNK2 was significantly decreased in the WNK2-inhibitor group (p<0.01), while the expressions of p-SHP2, p-AKT and p-ERK1/2 were significantly increased in the WNK2-inhibitor group (p<0.01). Compared with the NC PHPS1 group, the expression of WNK2 was significantly decreased in the WNK2-inhibitor PHPS1 group (p<0.01), while the expressions of p-SHP2, p-AKT, and p-ERK1/2 were reversed and had no significant differences compared with the NC PHPS1 group (p>0.05). The cell scratch assay and Transwell results showed that the migration and invasion ability of HepG2 cells in the WNK2-inhibitor group was significantly increased compared with the NC group (p<0.01). The migration and invasion ability of HepG2 cells in the NC PHPS1 group and WNK2-inhibitor PHPS1 group were significantly decreased with no significant difference (p>0.05). The results of the monoclonal proliferation experiment showed that the proliferation capacity of HepG2 cells in the WNK2-inhibitor group was significantly increased compared with the NC group (p<0.01), while the proliferation ability of HepG2 cells in the NC PHPS1 group and WNK2-inhibitor PHPS1 group was significantly decreased with no significant difference (p>0.05). Conclusion WNK2 inhibits the expression of SHP2 in HCC by regulating the level of oxidative stress, delaying the proliferation and migration of HCC.
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IL2rg-/- knockout rats support human RSV a prolonged infection

FAN CHANGFA, XIONG RUI, WU YONG, YANG YANWEI, QU ZHE, LIU SUSU, WANG YUYA, MA LIYING, FU RUI, PENG YIHONG, LIANG CHUNNAN

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Human respiratory syncytial virus (hRSV) is the second leading aetiology of lower respiratory infection deaths in the world. Disease animal models play an important role in the development of vaccine and therapeutic drugs, and a variety of animals, including non-human primates such as chimpanzees, have been made to develop models of hRSV infection. However, the limitations such as semi-permissiveness and short duration of infection have impeded their applications in both the pathogenesis of hRSV and therapeutics development. Here, we presented an hRSV infection model based on IL2rg gene deficient rat using TALEN technology. IL2rg-/- knockout rats can sustain high viral loads in nasal cavity upon intranasal inoculation with hRSV. The average peak titer rapidly reached 1?010 copies/g in nasal tissue, as well as 1?07 copies/g up to 5 weeks post infection. Since IL2rg-/- knockout rats is genetically heritable, with stable traits and easy production, which facilitate the standardization of disease models. Compared with mice, rats have the advantages of being larger and more convenient for collecting sufficient samples. This IL2rg-/- rat model can be used to study the transmission and pathogenesis of hRSV and is a useful tool for evaluating therapies.
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Methodological discussion based on preclinical studies of fecal bacteria transplantation in rodents

sunshiqi, Liu Lu, Hu Shuangyuan, Wang Yuyan, Sun Mingsheng, Zhao Ling

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Fecal microbiota transplant (FMT) is a therapeutic modality that targets intestinal microorganisms by transplanting fecal microorganisms from the donor into the recipient's gastrointestinal tract in order to reconstitute the recipient's intestinal flora. However, the mechanism of action and adverse effects of FMT in response to different diseases have not yet been clarified, and thus it cannot be widely applied in the clinic, and still needs to rely on in-depth preclinical studies. The highly inconsistent or incomplete experimental details in current reports, coupled with the lack of authoritative standards and recommendations, not only seriously affect the interpretation of their findings and the replication of experimental procedures, but also hinder the clinical translation of the results. We review and discuss the key steps of recipient selection and graft sample collection, storage, graft material preparation, and grafting route, with the aim of improving the utilization of experimental animals, consumables, and labor, and providing methodological recommendations and references to achieve replicability and standardization of FMT preclinical studies.
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Comparison of different Regimens in Isoprenaline-induced Chronic Heart Failure Models

Tanyuquan, huzhixi

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Objective: To compare animal models prepared by three different protocols and to explore a stable, reliable and reproducible mouse model of chronic heart failure. Methods: Twenty-five male C57BL/6J mice were randomly divided into four groups, namely blank group (ZC), model A group (MA), B group (MB) and C group (MC). The model group adopted different preparation protocols for continuous injection of ISO, in which the MA group and the MB group were dose-decreasing modeling, and the MA group (10 mg·kg-1 on the first day, 5 mg·kg-1 on the second day, 2.5 mg·kg-1·d-1 on the 3rd~30th day; subcutaneous injection for 30 days); MB group (20 mg·kg-1 on day 1, 10 mg·kg-1 on day 2, 5 mg·kg-1·d-1 on day 3~14; Subcutaneous injection for 14 days), MC group (constant dose 7.5 mg·kg-1·d-1, intraperitoneal injection for 28 days), CHF model was constructed. The day after the end of injection, the survival and mold formation rates of each group of mice were calculated. Cardiac function was measured by cardiac ultrasound and serum levels of N-Terminal Pro-Brain Natriuretic Peptide (NT-pro BNP), interleukin 6 (IL-6), and tumor necrosis factor (TNF-α) were measured in serum. Results: After the end of injection at the end of the fourth week, all model groups were effective in inducing CHF. However, based on the comprehensive test results, it was found that the mold-making situation of the MC group with a concentration of 7.5mg/kg was the most stable, which was more suitable for subsequent research on traditional Chinese medicine. Conclusion: ISO prepared mouse CHF model with constant 7.5 mg·kg-1·d-1 and continuous intraperitoneal injection for 28 days.
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Research advances on the use of pigeon in animal robot

Li Mengmeng, Yang Long, Yang Lifang, Wan Hong, Shang Zhigang

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Pigeons have the habits of clustering and homing, which are good at long-distance weight-bearing and continuous flight with excellent navigation and spatial cognitive abilities. Pigeons have been widely used in animal robot research in recent years. Pigeon robot achieves motor behavior control by applying neural information intervention to specific neural targets in the pigeon’s brain. According to the distribution of hierarchical multi-level neural regulatory targets in the pigeon’s brain, this review summarizes the research progress of pigeon robot based on the sensory system, motivation and emotional system or cortex and midbrain motor area respectively. This review aims to provide reference and guidance for further application research of pigeon robot in space perception, reconnaissance and anti-terrorism search and rescue.
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Analysis of the regulatory effect of Angelica dahurica on the MrgprD-TRPA1 pathway in neuropathic pain

Gu Leying, Yang Niuniu, Yu Kangying, Meng Yaqin, Song Shaozheng

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Objective Neuropathic pain was a kind of chronic pain caused by central or peripheral nervous system injury, dysfunction or transient disorder. Its mechanism was related to some receptors. In order to analyze the analgesic effect of Angelica dahurica in neuropathic pain and its regulation on MrgprD-TRPA1 signaling pathway. Methods Firstly, the CCI mouse model was prepared by using sterile surgical ligation and wrapping of the sciatic nerve in 30 mice. Secondly, the VonFrey experiment was used to detect the behavioral changes in pain induced by mechanical stimulation in mice, and the thermal radiation experiment was used to evaluate the thermal hyperalgesia of Angelica dahurica in mice. Then, the effects of Angelica dahurica on the expression of MrgprD and TRPA1 proteins, the number of DRG positive neurons, and the mRNA levels of MrgprD and TRPA1 in mice were detected by Western blot, immunofluorescence, and RT-PCR, respectively. Finally, the differences in fluorescence signal intensity were analyzed through calcium imaging experiments on HEK293 cells after single transfection and co-transfection of MrgprD and TRPA1 plasmids, respectively. Results A total of 25 CCI mouse models were successfully prepared, with a modeling rate of 83.33% (25/30). After the 7th day, CCI mice showed the most significant differences in mechanical stimulation threshold and thermal radiation foot contraction latency, and reached the lowest value.The mechanical threshold and foot retraction latency of CCI mice treated with Angelica dahurica were significantly higher than those treated with pure water and CCI mice not treated with Angelica dahurica (P<0.05). The expression levels of MrgprD and TRPA1 proteins in CCI mice treated with Angelica dahurica were significantly lower than those in CCI mice treated with pure water (0.73 ± 0.11 vs 2.69 ± 0.23, 0.42 ± 0.09 vs 2.03 ± 0.18, P<0.05). The number of MrgprD and TRPA1 positive neurons in DRG of CCI mice treated with Angelica dahurica was significantly lower than that of CCI mice treated with pure water (654 ± 47 vs 1162 ± 63, P<0.05). The relative expression levels of MrgprD and TRPA1 mRNA in CCI mice treated with Angelica dahurica were significantly lower than those in CCI mice treated with pure water and CCI mice not treated with Angelica dahurica(P<0.05). The fluorescence intensity in HEK293 cells co transfected with MrgprD and TRPA1 plasmids was significantly higher than that in single transfected and blank controls (P<0.05). Conclusion This study successfully established a mouse CCI model through ligation and winding, exploring the analgesic effect and mechanism of Angelica dahurica in the CCI model, and proving that MrgprD-TRPA1 is an important target for neuropathic pain. Angelica dahurica can inhibit the degree of neuropathic pain by regulating the signal transduction pathway of MrgprD-TRPA1, which laid a foundation for further research on the development of new clinical analgesic drugs and analgesic mechanisms.
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Mechanism of Icariin regulating NLRP3 inflammasome against Cerebral Ischemia Reperfusion

zengqi, WU Yachen, LIU Ming

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Objective: To investigate the therapeutic effect of Icariin on cerebral ischemia reperfusion injury in rats. Methods: The rat model of focal cerebral ischemia reperfusion was established by the method of thread bolt. After 24 hours，rats were randomly divided into sham operation group，model group，Butylphthalide group（70 mg穔g-1），and high，medium and low dose groups （80、40、20 mg穔g-1） of Icariin. The volume of gastric administration was 10 mL穔g-1 once a day for 13 d. The changes in neurological deficit symptoms of rats were detected by neurological function score，while the pathological damage of cerebral cortex were detected by HE，the expression changes of IL-1β and IL-18 protein were detected by Immunohistochemistry，the protein expressions of NLRP3，ASC and Caspase-1 in cerebral cortex were detected by Western blot. Results: Compared with the sham operation group，the neurological score of the model group increased. The pathological results showed that neurons necrosis or glial cell proliferation in different degrees could be seen in the marginal area of each group. The contents of IL-18 and IL-1β protein in the brain tissue of the model group increased significantly，and the expressions of NLRP3，ASC and Caspase-1 protein in the brain tissue of the model group increased significantly (P < 0.01, P < 0.05). After Icariin treatment，compared with the model group，the neurological function score of the treatment group was improved. The pathological results showed that the degree of neuronal necrosis was significantly reduced，and the contents of IL-1β and IL-18 protein in the brain were significantly reduced.；The expression of NLRP3，ASC and Caspase-1 protein in rat cerebral cortex decreased significantly (P < 0.01, P < 0.05). Conclusions: Icariin in the treatment of cerebral ischemia reperfusion injury may be related to regulation NLRP3 inflammasome.
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Effects of resistance exercise on lncRNA and mRNA gene expression profiles of SAMP8 mice skeletal muscle

FU ZETING, LI ZHONGHAO, LI LUNYU, LIU HONGZHENG, DING HAILI

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【Abstract】Objective To explore the potential regulatory mechanism of resistance exercise on senescence accelerated-prone mouse (SAMP8) by comparing the differential expression of lncRNA and mRNA in quadriceps between exercise group and control group by RNA-seq technology. Methods: Twenty-eight-week-old male SAMP8 mice were divided into a model group and resistance exercise group, with six mice in each group. Another eight SAMR1 mice of the same age were used as the control group. The resistance exercise group received 8 weeks of increasing weight climbing exercise training. The relative grip strength was performed every 1 weeks and the rotarod test was performed every 2 weeks. Hematoxylin-eosin staining was used to observe the histological changes of the right quadriceps femoris, and take the left quadriceps for RNA-seq (RNA-sequencing). The differentially expressed long non-coding RNA (lncRNA) and mRNA were screened. These Genes were then analyzed for enrichment by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Finally, the key differentially expressed genes were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) to verify the accuracy of RNA-seq results. Results: (1) Compared with the control group, the relative grip strength and rotarod test time of model group were decreased significantly(P < 0.01). After 8 weeks of resistance exercise, compared with model group, the relative grasping strength and the rotarod test time of resistance exercise groups were increased significantly(P < 0.01). (2) The results of Hematoxylin-eosin staining showed that compared with the C group, the cross-sectional area of muscle fibers in M group was significantly lower(P < 0.01), compared with the M group, the cross-sectional area of muscle fibers in R group was significantly increased (P < 0.01). (3) Through differential expression analysis, we found 182 upregulated and 218 downregulated lncRNAs, and 454 upregulated and 289 downregulated mRNAs in the comparison between the M group and the R group. The KEGG pathways of lncRNA target genes between the M group and the R group were significantly enriched in Intestinal immune network for IgA production、NF-kappa B signaling pathway、inflammatory bowel disease, etc. Conclusions: (1) This study demonstrated that resistance exercise can improve skeletal muscle function in SAMP8 mice with sarcopenia, and evaluated the differentially expressed lncRNA and mRNA in M group and R group by RNA-seq. These genes may be the target of sarcopenia therapy. (2) By analyzing the biological information of the target genes of differentially expressed lncRNAs and mRNAs, it is possible to further understand the mechanism of resistance exercise to improve sarcopenia.
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Research Progress in experimental models of Idiopathic Pulmonary Fibrosis

LI Zhi-hui, YU Xue-qing, yanshuguang, Yu Ning xia, 臧丹阳

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Idiopathic pulmonary fibrosis ( IPF ) is a chronic, progressive interstitial lung disease. The incidence of IPF is increasing year by year, with high mortality and poor prognosis. At present, the pathogenesis of IPF is still unclear, and its treatment measures are limited. The experimental model is a key tool for further studying the pathogenesis of IPF and exploring effective prevention and treatment measures. In recent years, its modeling methods have been continuously developed and optimized. In view of this, this study summarizes the construction methods and research progress of IPF experimental models in recent years, in order to provide ideas and references for preclinical research to select appropriate experimental models.
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Construction and evaluation of the orthotopic transplantation tumor model derived from transgenic mouse with spontaneous pancreatic cancer

AN Qingling, TAN Dengxu, ZHAO Ya, ZHANG Caiqin, SHI Changhong

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Objective To construct the orthotopic transplantation tumor model with pancreatic cancer derived from transgenic LSL-Kras^G12D/+, LSL-Trp53^{R172 H/ +}, Pdx1-Cre(KPC) mice. To provide a stable and reliable preclinical animal model for studying the development mechanism and treatment strategy of pancreatic cancer. Methods The tumor tissue derived KPC transgenic mice with spontaneous pancreatic cancer was transplanted into C57BL/6J mouse pancreas. Ultrasound detection was used to monitor the growth of tumor. H E staining and immunofluorescence staining were used to evaluate the pathological characteristics of this model. Results The tumor derived from KPC mice could grow steadily on the pancreas of C57BL/6J mice. Tumor proliferation index Ki67, matrix fibrosis marker αSMA, immune cell markers CD45 and CD206 were all stably expressed in the tumor. The model stably retains the pathological features of primary pancreatic cancer. Widespread tumor metastases were development in this model, which was similar to those observed in patients with pancreatic cancer. Conclusion The orthotopic transplantation model derived from transgenic mouse with spontaneous pancreatic cancer was established successfully. The model can simulate the stromal environment and immune cell infiltration of pancreatic cancer, and retains strong stability and uniformity with original tumor. It can be used as an effective preclinical model to study pancreatic cancer progression and treatment strategies.
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Based on TRL4/MyD88/NF-κB pathway to explore the protective effect of Periplaneta americana powder on rats with spinal cord injury.

Li jie, ZHOU Bang-yu, MA Yan-bo, RUAN Yu-shan, LI Shao-bo

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Abstract: Objective To explore the protective effect and possible mechanism of Periplaneta americana powder on rats with spinal cord injury. Methods Forty-eight male SD rats were randomly divided into sham operation group, saline group, periplaneta americana powder group, and Toll-like receptors-4 inhibitor group. Except for the sham operation group, the rat spinal cord hemitransection injury model was constructed in the other three groups. The sham operation group received no treatment after operation, the saline group and the periplaneta americana powder group were given intragastric administration of equal volumes of normal saline and periplaneta americana powder (630 mg/kg), and the TLR4 inhibitor group was given intraperitoneal injection of TLR4 inhibitor (3 mg/kg ) deal with. On the 1 d, 3 d, 7 d, and 14 d after the operation, the motor function of the rat hind limbs was evaluated by BBB score, the histopathological changes of the spinal cord were observed by hematoxylin-eosin staining, and the changes in the number of neurons were observed by immunohistochemistry. The expressions of inflammatory factors IL-1, IL-6, IL-10 and TNF-α were detected by ELISA, and the expressions of TLR4, myeloid differentiation factor 88 (MyD88) and NF-κB p65 were detected by Western blot. Results Compared with the sham operation group, the BBB score and the number of neurons in the saline group were significantly decreased, while the degree of pathological damage, IL-1, IL-6, TNF-α, TLR4, MyD88, and NF-κB p65 levels were significantly increased (P < 0.05), compared with the saline group, the periplaneta americana powder group and the TLR4 inhibitor group increased the BBB score, the number of neurons, decreased the degree of pathological damage, IL-1, IL-6, TNF-α, TLR4, MyD88, NF-κB p65 levels (P < 0.05), compared with the TLR4 inhibitor group, the periplaneta americana powder group better increased the BBB score, the number of neurons, decreased the degree of pathological damage, and the expression of IL-1 and TNF-α. Conclusions Periplaneta americana powder can reduce the production of inflammatory factors after spinal cord injury by inhibiting the TLR4/MyD88/NF-κB pathway, and play a role in protecting nerves and promoting motor recovery.
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Establishment of a mouse model of thoracic aortic dissection with acute lung injury by β-aminopropionitrile

maizhiyan, jiang li qing, zhuhanzhao, zhangliyun, wangyun, duanweixun

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【Abstract】Objective A feasible and stable mouse model of thoracic aortic dissection (TAD) combined with acute lung injury (ALI) was constructed using β-aminopropionitrile monfumarate (BAPN,1g/kg/d) administered by drinking water. The mouse model of TAD combined with acute lung injury (ALI) was constructed to provide a rational platform for the study of TAD combined with ALI. Methods Forty-five SPF-grade 3-week-old C57BL/6J male mice were selected and randomly divided into 15 mice in the CON group (normal dietary water) and 30 mice in the BAPN group (drinking water administration with sterile water configured as a solution of 1 g/kg/d) for 4 weeks. During the experimental period, the general condition and modeling rate of mice in the two groups were observed, and the TAD model of mice was validated and the BAPN group was divided into TAD and Non-TAD groups by measuring the maximum diameter of the ascending aorta and H&E staining of the aortic tissues of the mice. The pathological staining of H&E, wet/dry weight ratio (W/D) and total protein level in alveolar lavage fluid (BALF), and interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) expression in the alveolar lavage fluid (BALF) to validate the TAD combined ALI model of mouse. Results BAPN intervention significantly delayed the increase in body mass and water intake in mice. Compared with the CON and Non-TAD groups, the maximum diameter of the ascending aorta of mice in the TAD group was significantly thickened (P < 0.05), H&E staining of the aorta showed significant thickening of the aortic wall, reduction in the number of smooth muscle cells, and fracture and disorder of elastic fibers, and H&E staining of the lung tissues showed significant interstitial edema and inflammatory exudation accompanied by thickening of the alveolar wall and enlargement of alveolar lumen, and a significant increase in the pathological scores of lung injury (P < 0.05), and total protein level and expression of IL-1β, IL-6, and TNF-α in lung tissue W/D and BALF were also significantly increased (P < 0.05), while there was no significant difference in the above indexes between the other two groups. Conclusion A mouse model of thoracic aortic dissection combined with acute lung injury can be successfully established by BAPN drinking water administration.
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Establishment of the human α-synuclein nuclear localization signal transgenic mice

wei meng chen

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【Abstract】 Objective To establish a human α-synuclein nuclear localization signal transgenic mouse model and investigate the effects of α-synuclein nuclear localization on the behavior of mice. Methods Human α-synuclein nuclear localization signal and EGFP lentiviral vectors were constructed.The transgenic mice were created with the microinjection method. Using PCR and Western blotting methods to identify the genotypes and protein expression of the transgenic founder mice and their offsprings. The immunofluorescence was used to examine the localization of human α-synuclein in the mouse brain tissue . The behavioral changes of the transgenic mice were evaluated by the open field test, rotarod test, and O maze test. Results Transgenic mice with human α-synuclein nuclear localization signal were successfully established. Human α-synuclein was widely expressed in various tissues of mice and showed obvious nuclear localization. Further studies found that human α-synuclein nuclear localization signal transgenic mice had significant motor dysfunction, astrocyte proliferation and inflammatory response at 2 months of age and exhibited significant anxiety-like symptoms and reduced expression of the γ-aminobutyric acid(GABA) gene at 9 months of age, which persisted until 12 months of age. Conclusion A human α-synuclein nuclear localization signal transgenic mouse model has been successfully established, human α-syn is widely expressed in various tissues of the mice with evident nuclear localization. The mice exhibit significant motor dysfunction and anxiety-like symptoms. The successful establishment of this model provides a foundation for studying the role of α-syn nuclear localization in Parkinson's disease.
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Establishment and evaluation of a rat model of heart failure with preserved ejection fraction induced by compound factors

shiyujiao, Yang chen guang, Qiao wen bo, Liu yong cheng, Liu si yu, Dong guo ju

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【Abstract】 Objective To evaluate the characteristics of a rat model of heart failure with preserved ejection fraction (HFpEF) induced by combined factors and to investigate the correlation of myocardial strain parameters with myocardial hypertrophy and fibrosis. Methods Eight WKY rats and eight spontaneously hypertensive rats (SHR) served as control groups and were given normal feed until the end of the experiment. Thirty two SHR rats were equally divided into SHR+S, SHR+F, SHR+SF, and SHR+ combined groups. They were fed with high salt feed, high fat feed, high salt-fat feed, and high salt-fat-sugar feed in combination with streptozotocin intraperitoneally for 30 weeks, respectively. After modeling, heart weight/body weight (HW/BW), systolic blood pressure (SBP), and diastolic blood pressure (DBP) were measured; Echocardiography was performed to measure left ventricular (LV) end-diastolic internal diameter (LVIDd), LV anterior wall thickness (LVAWd), LV posterior wall thickness (LVPWd), LV ejection fraction (LVEF), isovolumetric diastolic time (IVRT), and peak early diastolic passive filling velocity (E) / early diastolic mitral annular velocity (e'); Speckle tracking echocardiography was conducted to determine global longitudinal strain (GLS) and strain rate (GLSr), global radial strain (GRS) and strain rate (GRSr), as well as global circumferential strain (GCS) and strain rate (GCSr); Serum is used to detect triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), glucose (GLU), and glycated serum protein (GSP); ELISA was used to examine serum B-type brain natriuretic peptide (BNP), angiotensin II (AngII),and Galectin-3 (Gal-3); Myocardium was subjected to HE and Masson staining for cardiomyocyte and myocardial fibrosis, and cardiomyocyte cross-sectional area (CSA) and collagen volume fraction (CVF) were calculated; In addition, the correlation of myocardial strain parameters with CSA and CVF was analyzed. Results Compared with the control group, in the model groups, especially in the SHR+Combined group, HW/BW, SBP, DBP, serum indexes (TC, TG, LDL-C, GLU, GSP, BNP, AngII, and Gal-3), and echocardiographic parameters (LVIDd, LVAWd, LVPWd, IVRT, and E/e') were significantly up-regulated; Absolute values of speckle-tracking echocardiographic parameters (GLS, GLSr, GRS, GRSr, GCS, and GCSr) were decreased considerably; HE and Masson staining of myocardial tissues suggested marked cardiomyocyte hypertrophy and fibrosis, and there were significant increases in CSA and CVF (P<0.01 or 0.05). Correlation analysis showed that GLSr, GCS, and GCSr were strongly linked to CSA; GLS, GLSr, and GCSr were strongly linked to CVF (P<0.01). Conclusions A rat model of HFpEF induced by hypertension and dysregulation of glucolipid metabolism replicated the basic characteristics of HFpEF in terms of etiology, clinical features, and myocardial pathologic changes and might be a reliable animal model of metabolic syndrome-related HFpEF. Moreover, myocardial strain indices are closely related to myocardial hypertrophy and fibrosis and might indirectly reflect subtle myocardial lesions and dysfunction.
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Effect of Qinji Liangxue mixture on similar animal models of hemorrhoids

ZHOU Mengen, LI Peng, JIN Wenqi, WANG Ruolin, GUO Xiutian

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Objective To explore the effect of Qinji Liangxue mixture (QLM) on similar animal models of hemorrhoid. Methods Animal models of acute hemorrhoid in rats were made by using croton oil preparation (COP) and glacial acetic acid respectively. Eighty SD female rats were randomly divided into control group, model group, low-dose QLM group (QLM-L), high-dose QLM group (QLM-H) and diosmin group (DOSM), with 8 rats in each group. Except for the control group, all the other groups were treated with COP or glacial acetic acid, and then treated with different drugs for 7 days. HE staining was used to observe the anorectal histomorphology induced by COP and glacial acetic acid, and ELISA was used to detect the levels of IL-8 and TNF-α in serum of rats induced by COP, and the area of perianal ulcer induced by glacial acetic acid was recorded. Twenty-four C57 mice were randomly divided into control group, QLM-L group, QLM-H group and adrenal chromazone tablet group (CT), with 6 mice in each group for 7 days. The hemostatic effect of QLM was evaluated by capillary method and tail cutting method. Results QLM can significantly improve the pathological injury of animal models with similar hemorrhoid diseases, reduce the score of pathological changes induced by COP (P < 0.05), reduce the levels of serum IL-8 and TNF-α(P < 0.05),, reduce the area of perianal ulcer induced by glacial acetic acid (P < 0.05),, and shorten the time of coagulation and bleeding(P < 0.05). Conclusions QLM has good anti-hemorrhoid activity, which may be achieved by anti-inflammation, hemostasis and reducing tissue damage.
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Effects of exercise regulating Nrf2/HO-1 pathway on improving HFFC diet-induced oxidative stress in hepatocytes

ping ye, zhang pei wen, yuan xin meng, xiang meng qi, yang meng fan, lin xiao xia, dong shi ru, liu yu ting, zhang yuan

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Objective To explore whether voluntary wheel running affects liver oxidative stress by regulating Nrf2/HO-1 pathway, thereby alleviating HFFC diet-related lipid deposition in liver. Methods 8-week-old C57BL/6J mice were randomly divided into normal diet group (NC group, n=10) and high fat, fructose and cholesterol diet group (HFFC group, n=20) after a week of adaptive feeding. Ten weeks of feeding later, mice in HFFC group were divided into quiet group (HFFC group, n=10) and HFFC combined with exercise intervention group (HFFC+EX group, n=10). HFFC+EX group mice were caged with voluntary running wheels for free movement, and the number of running wheels was recorded every day for 8 weeks. After the last intervention, the mice were sacrificed by fasting for 12 hours at an interval of 24 hours, and the blood and liver were taken for detection. Results (1) The body weight, liver weight and liver index of mice induced by HFFC diet were significantly higher than those of NC group, and significantly decreased after exercise intervention (P<0.05). (2) Compared with NC group, HDL-C and LDL-C in HFFC group were significantly increased, and LDL-C level was significantly decreased after 8 weeks of exercise intervention (P<0.05). (3) The liver fat drop area and liver TG content in HFFC group were significantly higher than those in NC group, while those in HFFC+EX group were significantly decreased (P<0.05). (4) Compared with NC group, the content of oxidase MDA and the expression level of HO-1 in HFFC group were significantly increased, and the nuclear translocation and gene expression of Nrf2 were significantly decreased. After exercise intervention, the activities of SOD and T-AOC were significantly decreased, and the nuclear translocation and gene expression of Nrf2, the expression levels of HO-1 and SOD-1 were significantly increased (P<0.05). (5) The number of hepatocyte apoptosis and CHOP expression in HFFC diet group were significantly higher than those in NC group, while the number of hepatocyte apoptosis, CHOP and Bax/Bcl-2 expression in exercise group were significantly lower than those in NC group (P<0.05). Conclusions Voluntary wheel running can alleviate liver lipid deposition induced by HFFC diet by regulating Nrf2/HO-1 pathway, thereby alleviating oxidative stress and reducing apoptosis in liver cells.
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Research progress on evaluation of diarrhea irritable bowel syndrome animal model

zhangjiahe, zhuwang, shendanting, yangxiling, liufengbin, houqiuke

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Irritable bowel syndrome (IBS) is one of the most common functional gastrointestinal disorders, of which diarrhea-predominant IBS (IBS-D) accounts for the largest proportion. The pathogenesis of IBS-D is complicated and diverse, and there is currently a lack of clinically effective drugs. The establishment of animal models is an essential tool for further studies of the disease mechanisms, evaluation of clinical efficacy, and drug development, and the preparation and evaluation standards of models are important factors affecting the quality of the research. Based on the currently accepted pathogenesis of IBS-D and the previous modeling experience of our research group, this review systematically summarizes the evaluation methods used in animal models of IBS-D in terms of diarrhea observation, visceral sensitivity tests, and intestinal motility tests, to provide a reference for future studies.
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Research progress on the construction and evaluation indicators of skin wound animal models based on physical methods

OU Xue, YU Zhijie, HE Yao, ZHENG Xiaoyuan

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Skin wounds are global public problems, especially in terms of the difficult healing of chronic wounds, which may seriously affect patients’ lives. Most skin-wound animal models are currently established by physical methods, and different animal models have different biological characteristics. This review therefore classifies mouse, rat, and other animal models of skin wounds according to the literature, summarizes and analyzes the construction of skin-wound animal models based on physical methods and evaluation indicators, and considers the advantages and disadvantages of different animal models, to provide a basis for the rational construction of skin-wound animal models and drug research and development.
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Screening and validation of age-related DNA methylation microhaplotypes in mouse blood

TIAN Yibo, WU Yujing, XIAO Junhua, ZHOU Yuxun, LI Kai

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Objective DNA methylation microhaplotype (DMH) refers to the combination of multiple methylation sites in a very short range, and its haplotypes are rich in diversity. Screening and validation of age-related DNA methylation microhaplotypes in mouse blood. Methods First, a theoretical dataset of DNA methylation microhaplotypes based on the mouse reference genome was constructed. Second, age-related DNA methylation microhaplotypes were screened by Spearman's rank correlation analysis using high-throughput sequencing information of DNA methylation in mouse blood from a network database. Finally, cross-validation was performed with a validation dataset. Results The number of DNA methylation microhaplotype sites within 50 bp in the mouse genome totaled 6,787,142, and DNA methylation microhaplotypes consisting of single-digit CpG sites accounted for 98.64 %. A total of 5,835 age-associated DNA methylation microhaplotypes were screened in 58 mouse blood samples (Spearman's rank correlation test, |rho| > 0.5, P < 0.01), and they accounted for 0.086 % of DNA methylation microhaplotypes. Finally, the top one hundred age-associated DNA methylation microhaplotypes with high correlation were validated in 95 independent samples, resulting in 44 loci. Conclusion The age-associated DNA methylation microhaplotypes screened in this study can be useful for future mouse blood apparent age prediction and aging studies.
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Analysis of animal models of vertigo based on the characteristics of clinical conditions in Chinese and Western medicine

liyaqing, wangxiaoyi, wangcan, miaomingsan

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【Abstract】Objective To classify and analyze the existing animal models of vertigo based on the clinical characteristics of Chinese and Western medicine, evaluate the clinical fit of the models and their advantages and disadvantages, in order to improve the existing animal models and provide a more intuitive reference for clinical research. Methods The existing animal models of vertigo in the database were searched, and the models were classified according to the modeling method, combined with the characteristics of Chinese and Western medicine clinical conditions of vertigo and the existing animal model evaluation methods, and assigned values to evaluate the clinical fit and the advantages and disadvantages of the models. Results The existing animal models of vertigo were neck surgery model, motor stimulation model, otogenic stimulation model, sclerotherapy injection model, flight variation pressure model, vertebral artery ligation model, and stasis injection model; among them, the Western medical fit was high (fit ≥ 60%) for neck surgery model (65%) and otogenic stimulation model (65%), and all models had moderate Chinese medical fit (50% ≤ fit < All models had moderate (50% ≤ 60%) or low (< 50%) TCM anastomosis, and there was no model with both high TCM and WCM anastomosis. Conclusion At present, the animal models of vertigo are mainly Western medicine disease models, and there is a lack of Chinese medicine evidence models, and there are few models with high clinical fit between Chinese and Western medicine, which fail to highlight the characteristics of Chinese medicine. Therefore, the construction of animal models of vertigo that are closely integrated with the clinical characteristics of Chinese and Western medicine can provide more reasonable and comprehensive experimental support for the development, screening and clinical evaluation of new anti-vertigo drugs.
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A mice model of hypothyroidism: comparison of two surgical procedures

Lan Haomiao, Che Hongying, Zhang Li, Mao Yu, Xie LinJun

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Objective Hypothyroidism is a serious endocrine disease and an independent risk factor for systemic diseases. Studies of hypothyroidism can't be done without establishing animal models. C57BL/6 and KunMing (KM) mice are ideal animal vectors for the study of endocrine and other diseases. However, there are few studies describing total thyroidectomy in mice in detail. Methods In this study, total thyroidectomy was performed in C57BL/6 and KM mice by ligation (operation method I) and hemostasis (operation method II), and the detailed operation process was recorded. Elisa was used to detect and compare the serum TT3, TT4 and TSH levels, body weight and neck tissue HE staining of mice before and after surgery to verify the model. Results The serum TT3 and TT4 were decreased (P< 0.05) and TSH was increased (P< 0.001) in the two model groups. The survival rate within 28 days after operation in group I and Group II was 40% and 60%, respectively. In KM mice, it was 50% and 40%. The body weight of the two groups of mice was significantly higher than that of the sham group. HE staining and microscopic observation showed that the cervical tissue of the two species of mice was thyroid tissue, and the back membrane of the thyroid was intact after isolation. Conclusion Both surgical methods can cause hypothyroidism in both types of mice. However, it is necessary to be familiar with the anatomical relationship of mouse thyroid gland and surrounding tissue, improve the proficiency of surgical operation, prevent the occurrence of postoperative hypocalcemia and infection, and thus improve the survival rate of mice after modeling.
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Analytical methodology of human umbilical cord mesenchymal stem cells in mouse tissues and their tissue distribution of NOG mice after single intravenous injection

Chen Guoyu, Pan Ruolang, Ye Zhichao, Shi Yuhua, Gu Liqiang, Xia Lijuan, Lin Xiaobo, Zhang Qiang, Xu Shasha, Shao Jinjin, Zhang Lijiang

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ABSTRACT: Objective To establish a quantitative PCR method for the analysis of human-derived SRY gene DNA in mouse tissues, and to study the tissue distribution of human umbilical cord mesenchymal stem cells in immunodeficient NOG mice after a single intravenous injection. Method A quantitative PCR method for the analysis of human SRY gene DNA in mouse tissues was established, and the methodological validation of standard curve and linear range, accuracy and precision, and stability was performed. Thirty-six NOG mice, one half of each sex, were given HUCMSCs 3.5?07 cells/kg by single intravenous injection, and six mice were anesthetized at 6 h, 12 h, 24 h, 72 h, 1 w and 2 w, respectively, and dissected after blood collection (EDTA anticoagulation). DNA was extracted from lung, kidney, heart, liver, brain, spinal cord, stomach, small intestine, fat, skin, spleen and testis (uterus and ovary), and the distribution of HUCMSCs in each tissue was determined by the validated method of quantitative PCR analysis of human-derived SRY gene in mouse tissues. In addition, 18 NOG mice, half males and half females, were divided into control group (6 mice) and treating group (12 mice), which were injected intravenously with 0.9% sodium chloride injection and HUCMSCs 3.5?07 cells/kg. The acute toxic reactions of mice were observed during the administration period, and four animals were dissected at 72 h, 2 w and 4 w after the administration to observe the gross organs, and the mitochondrial protein expression was detected by immunohistochemistry in paraffin sections of lung tissues to analyze the colonization of HUCMSCs in lung tissues. Results The established quantitative PCR method for human-derived SRY gene DNA in mouse tissues met the validation criteria for each index. After a single intravenous injection in NOG mice, HUCMSCs were mainly distributed in the lungs and bloodwithin 1w after administration, with higher concentrations in lung tissues than in blood, and the concentration of HUCMSCs in lung tissue and blood maintained relatively stable levels within 6h~24h and 6h~72h, respectively, and then decreased over time. The distribution of HUCMSCs was not measured in other tissues at all sampling points . The colonization results showed that HUCMSCs were detected in lungs 72h after intravenous injection, but not at 2w and 4w. No obvious acute toxicity was observed in NOG mice after single intravenous administration of HUCMSCs. Conclusion The established method for analyzing the distribution of HUCMSCs in mouse tissue is reliable and feasible. HUCMSCs were mainly distributed in lung and blood of NOG mice within 1w after single intravenous injection, and mainly colonized in lung tissue at 72h. The single intravenous administration of HUCMSCs has a good safety profile.
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Progress of animal model of bleomycin-induced pulmonary fibrosis

wangbo, songqinghua, tanghuimeng, liuyang, xieyang, tianyange

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Pulmonary fibrosis (PF) is a progressive, interstitial fibrotic lung disease characterized by persistent scar formation in the lung parenchyma and a reduced quality of life and poor prognosis for patients. The pathogenesis of PF is unknown and there is a lack of effective therapeutic agents, and animal models are the main tool to explore the pathogenesis of the disease and to find effective therapeutic agents. A variety of factors can induce fibrosis formation, and PF models can be induced to different degrees according to known etiology. Among them, bleomycin-induced models are widely used because of their reproducibility and similarity between fibrosis pathology and clinical conditions, and their induction methods mainly include intratracheal drip, intratracheal nebulization, tail vein injection, intraperitoneal injection and transnasal inhalation, and they are classified into single dose and multiple doses according to the frequency of induction. Based on the relevant literature, this paper summarizes the characteristics of the PF model established by BLM with different induction frequencies and induction methods, and provides a basis for the application of this model.
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Effects of Oral Probiotics on Gut Microbiota Structure in Subcutaneous Tumor of Colon Cancer Mice Based on 16S rRNA Sequencing Technology

Zhang Shuling, Wang Junwei, Zuo Luyu, Hu Shiliang, Sun Junzhi, Wang Chun

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Objective To investigate the effects of oral probiotics on gut microbiota diversity, colony structure, and intergroup differences in mice with subcutaneous colon cancer tumors, based on 16S rRNA sequencing technology. Method Twenty-four 6-week-old male BALB/c mice were divided randomly into normal control (NC, n=8), model (M, n=8), and probiotic+model groups (PM, n=8) after adaptive feeding for 1 week. Mice in the PM group were given 200 μL probiotic mixed solution (Bifidobacterium longum and Lactobacillus delbrueckii subsp. bulgaricus mixed lyophilized powder, 2×108 colony-forming units) by gavage three times/week for 7 weeks, while the M and PM groups received 200 μL normal saline. At 10 weeks old, 0.2 mL CT26.WT cell suspension (1×107/mL) was inoculated subcutaneously into the left hind limbs of M and PM mice, while NC mice were inoculated with 0.2 mL normal saline. The general condition of the mice and growth of subcutaneous tumors, and changes in the gut microbiota structure by 16S rRNA sequencing were monitored. Results The subcutaneous tumors of the M group were prominent, and the subcutaneous tumor volume and weight of the PM group were significantly reduced (P<0.05). Compared with NC group, Alpha diversity index was lower in the M group, and a significant difference of Beta diversity inter groups (P<0.01).And supplementation of probiotics had a certain effect on gut microbiota diversity in the M group. Compared with M group, the relative abundance of Bacteroidetes, Proteobacteria, Muribaculaceae, Bacteroides were higher in the PM group, while the relative abundance of Firmicutes, Desulfobacterota, Lachnospiraceae_NK4A136_group, Alistipes were lower in the PM group. LEfSe analysis showed that Muribaculaceae and Bacteroides in the PM group were different species with high abundance (LDA values >4). Conclusions Oral probiotics may improve the gut microbiota by increasing the relative abundance of beneficial Muribaculaceae and Bacteroides in subcutaneous tumors in mice with colon cancer.
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Advances in the application of 5XFAD transgenic mice in Alzheimer's disease research

liu yi duan, ye tian yuan

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5×FAD mice (Transgenic mice with five familial Alzheimer's disease) are APP/PS1 transgenic mice carrying five familial gene mutations. With beta-amyloid precursor protein (amyloid precursor protein, APP) related to the mutation of K670N/M671L (Swedish), 1716 v (Florida) and V7171 (London), and the old element - 1 (early presenilin 1, PS1) were MI46L and L286. 5×FAD mice had A large amount of β-amyloid (Aβ) in the brain at 1.5 months of age, and neuritic plaques (NP) began to appear at 2 months of age. The pathological phenotypes of 5×FAD mice included amyloid plaque aggregation, neuronal loss, gliosis, and memory dysfunction.The biological characteristics of 5×FAD mice may involve changes in the formation of brain Aβ plaques, hyperphosphorylation of Tau protein, synaptic dysfunction, neuroinflammatory response, mitochondrial dysfunction, blood-brain barrier injury, neuronal injury, endoplasmic reticulum stress and eye lesions. As a classic animal model of Alzheimer's disease, 5×FAD transgenic mice can simulate the neuropathological process and behavioral manifestations of AD patients in the late stage, and are widely used in the pathogenesis of AD and the development of new drugs for AD. In this paper, the model construction, biological background, biological characteristics of 5×FAD transgenic mice and the development and application of AD prevention and treatment drugs were summarized and reviewed, in order to provide reference for the application of 5×FAD transgenic mice in AD research.
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Effect of different sterilization methods on the nutritional composition of pig specific formula milk powder

GUO ya-xi, Liuruixue, Duxiaopeng, Zhuhua

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【Abstract】Objective Using different sterilization methods to sterilize pig specific formula milk powder, exploring the sterilization method and conditions that minimize the loss of nutritional components in formula milk powder. Method Pig-specific formula milk powder was divided into high-pressure sterilization and irradiation sterilization groups. Formula milk powder in the high-pressure group was sterilized using different sterilization conditions and that in the irradiation group was sterilized using different 60Co γ-radiation doses. The sterility and the nutritional contents of the sterilized formula milk powders were determined according to national standards. Results The sterility tests for both groups of formula milk powder were negative. Compared to control group, the crude protein contents were significantly lower in formula in the high-pressure group sterilized at 121℃ for 30 min and in the irradiation liquid group sterilized at 50 kGy (P < 0.01). The water, crude protein, and calcium contents were significantly lower (P < 0.001) in the irradiation group sterilized at 50 kGy. There was no significant difference in the valine, isoleucine, or leucine content under 50 kGy sterilization conditions in the irradiation sterilized group, but all amino acid contents were decreased in the high-pressure sterilization and irradiation sterilized liquid groups (P < 0.001). Analysis of trace elements showed an increased iron content (P < 0.001) in formula sterilized at 121℃ for 30 min in the high-pressure sterilization group, increased iron and potassium contents (P < 0.001) under 25 kGy sterilization conditions in the irradiation sterilization liquid group, and increased magnesium content (P < 0.01). The magnesium (P < 0.05) and sodium contents (P < 0.01) differed significantly in formula treated under 50 kGy sterilization conditions in the irradiation sterilized powder group. Vitamin E and Vitamin B2 contents were increased in formula sterilized at 121℃ for 30 min in the high-pressure sterilization group (P < 0.001), the Vitamin E content was increased (P < 0.05) and the Vitamin B2 content was decreased (P < 0.001) in formula sterilized under 50 kGy conditions in the irradiation sterilization liquid group, and the Vitamin E and Vitamin A contents were decreased in formula sterilized at 25 kGy in the irradiation sterilized powder group (P < 0.001). Conclusion Sterilization at 121℃ for 30 min resulted in the least loss of nutritional components in the high-pressure sterilization group, while irradiation sterilization resulted in the least loss of nutrients at a dose of 50 kGy. Comparing the two sterilization methods, irradiation of milk powder at 50 kGy resulted in the least loss of nutrient content.
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Research progress of pig in biology as model animal

wangyan, Xing Kai, Cao Yong Chun

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Animal models of human diseases include experimental animals and related materials established during biomedical research, which in turn play a vital role in medical research. Pigs and humans are similar in terms of their anatomy, physiology, immunology, and genetics. Pigs are thus suitable model animals for biomedical research and have various advantages compared with other model animals. Recent advances in biotechnology, such as genetic engineering, have contributed to a rapid increase in the use of pig models for human disease research. In addition to serving as xenotransplant organ donors and as tools in drug-design studies, pigs can also be used as model animals to study human developmental processes, congenital diseases, and disease-response mechanisms, thus making important contributions to improving human health. This review considers the current status and future applications of pigs as research models for studies of human cardiovascular diseases, cancer, ophthalmology, craniofacial, musculoskeletal, and skin research, reproductive and fetal development, nutrition, microbiome research, brain and neurodegenerative diseases, diabetes, infectious diseases, and vaccine design, as well as for xenotransplantation.
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Syndrome evaluation of hot flash model rats induced by bilateral oophorectomy and tamoxifen administration

Xie hiana, Pan zhiqiang

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【Abstract】Objective To explore the animal model of syndrome used in the study of hot flash phenomenon in women. Methods Twenty-four female SD rats were randomly divided into three groups: sham operation group, bilateral oophorectomy group and tamoxifen group, with 8 rats in each group. Two kinds of hot flash animal models were induced by bilateral oophorectomy and tamoxifen intragastory at 10 mg/kg/d, respectively. On the 14th and 28th days of modeling, the open field activity, anal temperature and body surface infrared thermogram of rats were detected. On the 29th day, the rats were killed, and the uterus was weighed and pathological sections were made. Blood estradiol and epinephrine were determined by ELISA. The gene expressions of adrenal sex hormone synthetase (Star, Cyp11a1, Cyp17a1, Cyp19a1, Por, Hsd3b2, Hsd17b1) and catecholamine substance synthetase (Th, Ddc, Dbh, Pnmt) in adrenal medulla were detected by RT-qPCR. Results Compared with sham operation group, the body weight of rats in bilateral ovariectomy group increased significantly (P < 0.01), while the body weight of tamoxifen group increased slowly. In the bilateral ovariectomies group, the maximum body surface temperature was significantly decreased on day 28 (P < 0.01), the difference between the maximum temperature and the minimum temperature in the abdomen was significantly increased on day 14 (P < 0.05), the difference between the maximum temperature and the minimum temperature in the back was significantly increased on day 28 (P < 0.01), and the open field activity of the rats was decreased (P < 0.01). The maximum body surface temperature of the tamoxifen group was significantly decreased (P < 0.01), but the open field activity of the rats was increased (P < 0.01). The uterine index of rats decreased significantly in both models (P < 0.01). Compared with sham operation group, E2 in bilateral oophorectomy group and tamoxifen group was significantly decreased (P < 0.01), NE and Epi were significantly decreased (P < 0.05), β-EP in bilateral oophorectomy group was also significantly decreased (P < 0.05). Compared with sham operation group, the adrenal Cyp11a1 gene expression in bilateral ovariectomized rats was significantly increased (P < 0.05), while the Cyp17a1 and Hsd17b1 gene expressions were significantly decreased (P < 0.05). The gene expressions of Star and Por in tamoxifen group were significantly increased (P < 0.01), while the gene expression of Cyp17a1 was significantly decreased (P < 0.01). Pnmt gene expression was significantly down-regulated in bilateral oophorectomy group (P < 0.01). conclusion Bilateral ovariectomized rats can be used for the study of perimenopausal hot flashes, whose syndromes are similar to those of kidney Yang deficiency and Yin deficiency in traditional Chinese medicine.
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Transcriptomic analysis of C57BL/6N-Tg(1.28HBV)/Vst Hepatitis B virus transgenic mice

li rongrong, sunxin, huangkai, zhaozhimin, pengyuan, liuchenghai

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【Abstract】Objective To observe the characteristics of C57BL/6N-Tg (1.28mer HBV)/Vst transgenic Hepatitis B virus (HBV-Tg) mouse model and analyze the transcriptomic characteristics of HBV-Tg mouse model. Methods 10 male HBV-Tg mice were used as experimental group and 10 wild-type mice as control group. The level of HBV DNA, HBsAg, HBeAg in serum and the expression of HBsAg and HBcAg in liver tissue were used to evaluate the virological characteristics of the model mice. The levels of serum ALT, AST, HE, Sirius red staining and Hyp in liver tissue were detected to evaluate the degree of liver inflammation and fibrosis. Liver tissue samples from 3 mice in each group were randomly selected for RNA extraction for high-throughput transcriptome sequencing. The significantly expressed differential genes were obtained by R software analysis, the functional enrichment of differential genes was obtained by GO and KEGG analysis, and then the genes with significant differences were verified by real-time fluorescence quantitative PCR (qRT-PCR). Results Compared with normal group, ALT and AST levels in model group were increased, and ALT was more significant (P<0.05). HE staining of liver tissue in model group showed the enlargement of liver nucleus and swelling of some hepatocytes. The results of Sirius red staining showed that there was a small amount of collagen deposition in the sink area and interlobule of HBV transgenic group, which was in the shape of thin lines. A total of 1352 differential genes were obtained by screening conditions (logFC>2x and P<0.05), including 703 up-regulated genes and 649 down-regulated genes. KEGG analysis suggested that differential genes were mainly enriched in PPAR signaling pathway, retinol metabolism, fatty acid degradation and other pathways (P<0.05). Significantly up-regulated differential genes mainly included Cyp4a10, Cyp4a14, Acot1, Acot3, Ehhadh, etc. Significantly down-regulated genes included Adh4, dnajb11, hspa5, scn5a, apol10b, etc. The trend was consistent after qRT-PCR detection (P<0.05). Conclusions HBV-Tg mice have a tendency of spontaneous fibrosis.Transcriptomic analysis shows that the potential mechanism of CHB mainly involves PPAR signaling pathway, retinol metabolism,fatty acid degradation,drug metabolism and other pathways.
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Research new progress in aging models of rodent animal

LI Xuechan, HAN Le, WANG Xuewen, LIU Lijun, WANG Jing

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Aging is a process of degenerative change that occurs as a result of time-related accumulation, associated with age-related diseases. Understanding the causes and mechanisms of aging and finding drugs that can effectively delay aging and prevent and cure age-related diseases currently present a great challenge for humans. Aging animal models thus represent an important tool in aging research, and various aging animal models have been created using different aging mechanisms. These different models having specific advantages and disadvantages, making them suitable for different research purposes. This review considers aging rodent models to provide information for aging research.
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Construction and evaluation for chemotherapeutic phlebitis rats induced by vinorelbine through dorsalis pedis vein

王, jihaijie

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To establish and evaluate for chemotherapeutic phlebitis rats induced by vinorelbine through dorsalis pedis vein. Methods The rats were randomly divided into the control and model group. Rats injected 0.1ml normal saline through the dorsalis pedis vein of the hind limb served as the control. While the rats in model group were injected with different concentrations of vinorelbine (2-5mg/ml) as above. General observation was performed and the hind limb volume was measured daily to calculate the swelling rate for 7 consecutive days. Then the rats were killed to collect the dorsalis pedis vein, and the histological changes were observed by HE staining. The microstructure changes on the surface of the vascular endometrium were observed by scanning electron microscopy. Results Rats injected with 2-5 mg/ml vinorelbine via dorsalis pedis vein could significantly induce hind limb swelling. The swelling rate increased with increasing concentration, and the peak of each group is reached on 3rd day. On the 7th day, the phlebitis rate in the 2 mg/ml group was 50%, while in the 3 mg/ml group of 83.3%. The 4 mg/ml group and 5 mg/ml group were all sucessful with grade Ⅲ accounting for 66.7% and grade Ⅳ accounting for 83.3%, respectively. Histopathology showed inflammatory cell infiltration, wall thickening, lumen stenosis and thrombosis in the surrounding tissues of veins. Scanning electron microscopy also showed that the tight junction of venous endothelial cells were destroyed, and the surface of the vascular lining were rough that resulting in blood cells adhesion. Conclusion Rats injected with 0.1ml 3-5mg/ml vinorelbine through the dorsalis pedis vein could induce red, swollen and cord like veins in lesions. Inflammatory cells infiltration around the vein, thickened vein wall, lumen stenosis and thrombosis were seen. Also, the surface of the vein intima was rough and adhered to a large number of blood cells. All of those were consistent with clinical chemotherapeutic phlebitis in terms of symptoms and pathological structure.
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Establishment and evaluation of a rat model of coronary microvascular disease with qi deficiency and blood stasis syndrome

kangjing, yang li li, wang ziyan, You yue, shiyue, ma yanlei, meng hongxu, lilei

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Objective To explore the establishment and evaluation of a rat model of coronary microvascular disease with Qi deficiency and blood stasis syndrome. Methods 45 male SD rats were randomly divided into sham operation group, coronary microvascular disease group and coronary microvascular qi deficiency and blood stasis syndrome group, with 15 rats in each group. The rats in the group of coronary microvascular disease with Qi deficiency and blood stasis syndrome were randomly deprived of sleep for 14 to 16 hours every day for 6 weeks, and the model of Qi deficiency syndrome was established. The animals in the sham operation group and the coronary microvascular disease group were fed normally for 6 weeks. Six weeks after the experiment, rats in the coronary microvascular disease group and coronary microvascular qi deficiency and blood stasis syndrome group were anesthetized, and then opened their chest and injected embolic microspheres (40~120μm) into the left ventricle,The animals in the sham operation group underwent thoracotomy without injection of embolic microspheres. On the 7th day after operation, relevant detection indicators were measured in each group. Results In the group of coronary microvascular disease, microsphere embolism was found in coronary microvascular, left ventricular ejection fraction and left ventricular shortening rate were significantly decreased, and the activities of creatine kinase MB isoenzyme (CK-MB) and lactate dehydrogenase (LDH) were significantly increased. The heart function, hemorheology, myocardial enzyme index and the degree of myocardial cell damage in the coronary microvascular Qi deficiency and blood stasis syndrome group were significantly different from those in the sham operation group. Conclusion Sleep deprivation combined with intraventricular injection of embolic microsphere can successfully establish a rat model of coronary microvascular disease with qi deficiency and blood stasis syndrome, which is more suitable for the study of pathogenesis and mechanism of traditional Chinese medicine
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The ferroptosis pathway mediated by GSH/GPx4 might play an important role in preventing hepatocyte peroxidative injury by aerobic exercise in elderly mice model

Liu Yuqian, Yang Wenqian, Wang haitao

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Objective To elucidate the role of Glutathione (GSH)/Glutathione peroxidase 4 (GPx4) -mediated ferroptosis pathway in preventing age-related hepatocyte peroxidation injury by aerobic exercise in mice, and to provide a new target for improving liver aging and metabolism disorders. Methods 20 SPF C57BL/6 male mice aged 52 weeks were randomly divided into the elderly control group, (EC) and the elderly exercise group (EE), with 10 mice in each group. The mice performed 16 w moderate intensity exercise with the incremental load (1~2 w 14 m/min, 3~4 w 15 m/min, 5~10 w 16 m/min, 11~16 w 17 m/min, 60 min/d, slope 0?. After perfusion of the ascending aorta, the lateral lobes of the liver were harvested for HE sections and ultrathin transmission electron microscope sections. The levels of 8-hydroxy-2 deoxyguanosine (8-OHdG), 4-Hydroxynonenal (4-HNE) in liver and serum interleukin-6 (IL-6) were detected by ELISA. Hepatic glycogen, triglyceride (TG), malondialdehyde (MDA), nicotinamide adenine dinucleotide phosphate (NADPH), and glutathione (GSH) were determined by colorimetry. Hepatic GPx4, glucose transporter (GLUT2) and NAD(P)H:quinone oxidoreductase 1(NQO1) and solute carrier protein 7 family member 11 (SLC7A11) were detected by Western blot. Results ① The oxidative damage of hepatocytes in elderly exercise mice were effectively delayed, and the normal structure of mitochondria and glycogen storage in hepatocytes were maintained. ② Compared with the elderly control group, the content of hepatic GSH and NADPH in the elderly exercise group were increased significantly (P<0.01). ③ Compared with the elderly control group, 8-OHdG, 4-HNE, MDA and non-heme iron in liver of the elderly exercise group were decreased significantly (P<0.01). ④ The expression of GPx4, NQO1 and SLC7A11 in the liver of the elderly exercise group were increased (P<0.01), while the expression of NOX2 was decreased (P<0.01). Conclusions The synthesis of GSH was increased in aged mice after aerobic exercise, which provided sufficient reaction substrates for GPx4, and GSH /GPx4 pathway was activated. The ferroptosis process was inhibited, which improved hepatocyte peroxidation damage caused by aging, and maintained the normal structure and physiological function of hepatocytes.
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Role of monoamine oxidase A (MAOA) in prostate cancer progression

CHEN Hanmu, LI Hui, ZHAO Jumei, SHIChanghong

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Monoamine oxidase A (MAOA) is a mitochondrial enzyme that catalyzes the oxidative deamination of monoamine neurotransmitters and dietary amines. It plays a crucial role in the pathogenesis, progress, and treatment of neuropsychiatric disorders. Recent studies have revealed that elevated expression of MAOA in prostate cancer (PCa) is closely associated with tumor progression and drives the heterogeneity of PCa. In this review, we summarize the role of MAOA in the development of PCa in different disease stages, including oncogenesis, development, invasion, metastasis, and drug resistance. We also discuss the involvement of MAOA in the tumor microenvironment and explore the potential utility of MAOA inhibitors. We further propose therapeutic strategies based on targeting MAOA in preclinical models to promote relevant clinical trials. This review aims to provide new potential therapeutic targets for the treatment of PCa.
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Research progress on the application of zebrafish in breast cancer

ZHANG Min, ZHANG Jingjing, NING Guozhu

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Zebrafish xenograft model is of great importance in cancer modelling, especially xenografting of breast cancer. It facilitates real-time observation of tumor cell growth, metastasis, and interactions with the immune system, thus providing novel insights and experimental foundations for breast cancer treatment. Furthermore, the zebrafish xenograft model offers a valuable tool for high-throughput drug screening. This review provides an overview of how zebrafish xenograft model contribute in elucidating the underlying mechanisms in breast cancer development, as well as their use in screening anti-tumor drugs and conducting therapeutic research.
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Establishment and evaluation of anterior cervical discectomy fusion optimization model in small tailed Han sheep model

DOU Xinyu, LIU YU, LIU Xiao, ZHU Bin, JIA Fei, WANG Linbang, SHEN Fei, LIU Xiaoguang

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Objective Cervical disc herniation (CDH) is one of the common orthopaedic diseases. With the in-depth study of it and the development of cervical implants, the establishment of cervical fusion animal models has become an indispensable part. At present, there are few reported researches about the establishment and evaluation of cervical fusion animal models in China. The aim of this study is to provide a good animal model and evaluation scheme of implants for cervical spine-related research. Methods Small tailed Han sheep were chosen for anterior cervical discectomy fusion (ACDF) after modified operation and polyetheretherketone (PEEK) interbody fusion cage (Cage) (control group), 3D-printed Ti6Al4V Cage (experimental group 1) and new method Ti6Al4V Cage (experimental group 2) were implanted in different cervical segments (C2/3-C4/5) of each sheep respectively. After surgery, hematology and histopathological analysis were performed to evaluate the recovery of sheep and the biosafety of the materials. X-ray, CT, Micro-CT and quantitative analysis, hard tissue section staining and biomechanical tests were executed to assess the conditions of bone in-growth and bone fusion. Results The modified ACDF ovine model was established successfully. There was no significant difference in the important indexes of hematology (P< 0.05), and histopathological analysis showed there were no pathological changes such as inflammatory cell infiltration. The implants had a good biosafety..Furthermore, X-ray and CT showed the position of internal fixation and the interbody fusion were good. The results of Micro-CT and quantitative analysis at 3 and 6 months after operation showed that compared with PEEK Cage group, the bone volume/total volume and trabecular number in the new method Ti6Al4V Cage group and 3D-printed Ti6Al4V Cage group were significantly increased (all P < 0.01), while the trabecular spacing decreased significantly (all P < 0.01). Moreover, the new method Ti6Al4V Cage group had more bone growth (P < 0.01). Hard tissue section staining demonstrated that the pores of the new method Ti6Al4V Cage and 3D-printed Ti6Al4V Cage had obvious bone growth and were relatively dense, and the combination was slightly better than that of PEEK Cage. Biomechanical evaluation indicated that compared with PEEK Cage, the new method Ti6Al4V Cage and 3D-printed Ti6Al4V Cage reduced the range of cervical flexion-extention , lateral bending and axial rotation to a certain extent (all P < 0.05), at the same time enhanced the stability of cervical vertebra, and the new method Ti6Al4V Cage was more advantageous (all P < 0.05). Conclusions After the establishment of the modified ACDF ovine model, reasonable and effective assessment methods were used to demonstrate the rationality and effectiveness of the model, and good biosecurity of the Cages of the three materials. Compared with PEEK Cage, the new method Ti6Al4V Cage and 3D-printed Ti6Al4V Cage had better performance in bone growth and bone fusion, which could enhance the stability of cervical vertebrae, and the new method Ti6Al4V Cage had more advantages.
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Establishment and Evaluation of Constipated Irritable Bowel Syndrome Model with Liver Depression Syndrome

He Jiahui, He Jieying, Zhang Bairong, Zhang Shuidi, Wei Yuanjun, Yang Chaoyan, Chen Yanfen

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Objective Through single factor modeling method and multi factor combined modeling method, to prepare the rat models of liver stagnation syndrome constipation type irritable bowel syndrome (IBS-C), and provide ideal experimental animal models of IBS-C by comparing different indicators. Methods 42 sprague-dawley rats were randomly divided into blank group (Normal group), cold water gavage group (Cold group), restraint group (Restrain group), tail clipping group (Tail group), cold water gavage+restraint group (C+R group), and cold water gavage+tail clipping group (C+T group). During the modeling period, the body weight, food intake, water intake, and survival status of each group of rats, and open field behavior, fecal Bristol score, visceral sensitivity, and small intestine propulsion were observed. HE staining was used to observe the pathological changes of the rat colon, and ELISA method was used to detect the content of 5-HT and VIP in serum and colon. Results After modeling, the weight loss of rats in each model group decreased (P < 0.05 or P < 0.01), the amount of food and water decreased, and the content of 5-HT in serum of each model group increased. In the Cold group, the number of fecal particles and Bristol score decreased, while the content of 5-HT in the colon increased (P < 0.05 or P < 0.01); the total distance and average speed of the restraint group in the open field decreased (P < 0.01); the preference for sugar water in the Tail group decreased (P < 0.01); in the C+T group, the preference rate for sugar water, total open field distance, small intestine propulsion rate, defecation particles, and Bristol score decreased, while the content of 5-HT in the colon increased and the VIP content decreased (P < 0.05 or P < 0.01); the total distance, average speed, and VIP content in the colon of the C+R group decreased (P < 0.05). Except for the Tail group, all other model groups showed visceral hypersensitivity (P < 0.05 or P < 0.01) at various pressure values on the 7th and 14th day of modeling; pathological observation showed that no significant inflammatory cell infiltration or pathological changes were observed in each model group. Conclusions The combination of ice water gastric lavage and tail clamping method can successfully establish a rat model of liver depression syndrome in IBS-C, which maybe the first choice of five methods and lays the foundation for systematic and in-depth research on the mechanism of traditional Chinese medicine in preventing and treating IBS-C.
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Research progress of candidate pathogenic genes related to depression

Li Mengyao, Gao Feng, Zheng Fanfan, He Mengze, He Zhao, Li Youlei

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Depression is a complex mental disease with polygenic inheritance and a high incidence. Our understanding of the clinical manifestations and pathogenesis of depression has recently improved. Continuous progress in gene-editing technologies has increased the construction efficiency and reduced the cost of gene-knockout animals, leading to their increasing use in the fields of basic research and drug development for depression and providing a powerful tool for revealing the pathogenesis of depression. In this review, we summarize recent progress in understanding the roles and mechanisms of candidate genes in depression using knockout model mice.
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Prox-1 induces new lymphatic vessels to participate in the repair of spinal cord acupuncture injury

chendandan, Meng fanwei

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Objective To observe the distribution and characteristics of lymphatic vessels in normal and injured mouse spinal cord, and to explore whether lymphatic vessels participate in the repair of spinal cord injury. Methods Adult male Kunming mice were randomly divided into two groups (n=36), the normal group did not damage the spinal cord, the experimental group used acupuncture to prepare a spinal cord injury model. The distribution of lymphatic endothelial cells（LECs） in the spinal cord was detected by immunohistochemistry, and the expressions of lymphatic endothelial cell markers Prospero-related homobox-1 (Prox-1), lymphatic vessel endothelial cell hyaluronic acid receptor-1 (LYVE-1), flat foot protein (podoplanin) and the vascular endothelial cell marker CD34 in the spinal cord of normal and acupuncture-injured mice were observed. The spinal cord samples were subjected to immunofluorescence staining, and the source of new LECs was explored by observing the co-expression of LYVE-1/prox-1, LYVE-1/podoplanin and CD34/Prox-1. Results Lymphangoid structures are present in the spinal cord of normal adult mice, and are distributed in segments, walking laterally between white matter and gray matter; nascent lymphangoid-like structures appear in the spinal cord at the site of acupuncture injury, and Prox-1, podoplanin, LYVE-1, and CD34 are expressed simultaneously, and the expression of Prox-1 is time-ordered; after scarring at spinal cord injury, the nascent lymphangoid-like structure disappears. Conclusion: Segmental, transversely distributed lymphangoid-like structures are present in the spinal cord of normal adult mice, and the neonatal lymphangoid-like structures are involved in the reconstruction of spinal cord injury, and the nascent lymphatic endothelial cells may originate from the surrounding existing lymphatic vessels or vascular endothelial cells.
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Establishment of a novel composite rat model of chronic skeletal muscle injury

lujingjing

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:The purpose of this study was to establish a new composite rat model of chronic skeletal muscle injury to explore the mechanisms and treatment methods of skeletal muscle injury. The model was established by combining vertical impact with prolonged forced sitting, and significant pathological changes and physiological dysfunction were observed. The model group rats had significantly lower open-field distances than the blank group, and HE staining showed muscle atrophy and rupture, as well as infiltration of inflammatory cells. The volcano plot in the sequencing results suggested significant differences in the Pf4 gene. The muscle tissue Go analysis of the model group rats showed upregulation of the oxygen binding and oxygen carrier activity pathways. KEGG analysis suggested that the chemokine signaling pathway and AMPK signaling pathway were activated. In conclusion, this study successfully established a composite model of skeletal muscle injury in rats, which can be used to study the pathological changes of skeletal muscle injury and evaluate the therapeutic efficacy of treatment methods, with practical application value.
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A Brief Review of the cultivation and utilization of the rat resources

Zhang Leying, Xu Wei, Zuo Qin

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Rats are one of the representative experimental animals since the genetic and environmental effects is comparatively easy to control and standardize. The rat is rather suitable as model for physiology, neuroethology, pathology and toxicology due to an approximately ten times larger body size compared to a mouse, offering several unique advantages in surgical procedures and clinical sampling of blood, tissues, etc. This paper review the cultivation and utilization history of the rat resources abroad to provide references of collecting, sharing and utilizing laboratory rat resources in our country.
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NK cells and the application of their immunotherapy in tumor immunity

wuxueliang

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Abstract Natural killer cells (NK) are important immune cells that recognize and eliminate virus-infected cells and tumor cells. NK cells play a crucial role in anti-tumor immunity by modulating the immune response through a variety of cytotoxic mechanisms and cytokine production. This article reviews the development and classification of NK cells, the mechanism of action, and the application of NK cell-based immunotherapy in tumor immunity, and clarifies the principle, status quo and development trend of NK cell-based anti-tumor immunotherapy, in order to provide ideas for the future research and development of NK cells.
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Analysis of glaucoma animal models based on data mining

yuanxifan, miaomingsan

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Objective ：To study the application of animal model of glaucoma and provide reference for the improvement of animal experimental methods and model. Methods Methods With“glaucoma ”and “animal models” as the main topics, literature related to animal models of glaucoma from 2012 to 2022 was collected from CNI and PubMed, and the species, gender, modeling methods and detection indexes of experimental animals were summarized, and the database was established for statistical analysis.Results A total of 400 articles conforming to the criteria were selected. Most of the experimental animals were C57BL / 6J mice, and most of them were male. Most of the modeling methods were anterior chamber injection induced type, transgenic type and laser photocoagulation induced type. The high frequency detection indicators mainly included intraocular pressure measurement, histopathology, western blot analysis and immunohistochemistry. Conclusions: at present, there are many methods to construct glaucoma animal models, but the intervention of related TCM factors is less. It is suggested to increase the glaucoma animal model combined with disease and syndrome. In this study, different animal models were evaluated by mining and analyzing the animal model experiments of glaucoma, and the mining contents provided references for the construction of animal models with high success rate of modeling, good reproducibility and high clinical coincidence, and provided ideas for the model improvement.
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Exploring the protective effect of Helleborus thibetanus Franch alcohol extract on bone destruction in CIA rats based on OPG/RANK/RANKL signaling pathway

shanwenting, liuxia

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【Abstract】 Objective To investigate the anti-inflammatory effect of Helleborus thibetanus Franchon on collagen induced arthritis (CIA) model rats and the effect on OPG/RANK/RANKL signaling pathway. METHODS Sixty female wistar rats were divided into (1) normal group (Control), (2) model group (Model), (3) positive drug group (methotrexate, MTX), (4) low-dose group, (5) medium-dose group, and (6) high-dose group. The CIA rat model was established by injecting bovine type II collagen into the tail root of rats using the collagen antibody induction method, and the drugs were administered by gavage after the model was successfully established. Normal group: 10 mL/(kg·d) of saline was given; model group: 10 mL/(kg·d) of saline was given; the positive drug group was given 2.0 mg/(kg·d) of MTX each time, three times a week; the iron chopsticks low, medium, and high dose groups were given 0.25g, 0.5g, 1.0g/(kg·d) each time; continuous gavage treatment for 25 day. Body mass of rats was recorded to observe the degree of foot swelling;rat ankle arthritis index score;micro-CT observation of histopathologic changes in ankle joint bone; hematoxylin-eosin (HE) staining for pathological changes in ankle joint bone tissue and synovial membrane of the rats;tests to observe changes in the number of osteoclasts were performed with anti-tartaric acid phosphatase (TRAP) ; PCR to detect the mRNA levels of osteoprotegerin (OPG), nuclear factor-κB receptor activator (RANK), RANK ligand (RANKL), tumor necrosis factor (TNF-α), and bone-forming protein 2 (BMP-2);Relative expression of OPG, RANK, RANKL, TNF-α and BMP-2 proteins were detected by protein immunoblotting (Western blot)Helleborus thibetanus Franchon on rats with rheumatoid arthritis and its potential pharmacological mechanism for osteoarthritic protection. Results The statistical results of the data showed that compared with the normal group, CIA rats showed slower growth of body mass (P<0.05), increased thickness of the plantar foot (P<0.05), narrowing of the joint cavity in the ankle joint, gnawing-like bone destruction of the bone tissues, and pathologic changes in the synovium,Such as inflammatory cell proliferation and abnormal synovial hyperplasia infiltration; Micro-CT results showed that: compared with the normal group, the model group, low and medium dose group, the ankle joint surface was uneven, incomplete in shape, and the ankle joint surface showed gnawing-like bone destruction, and the high-dose group had no significant changes in all indexes compared with the normal group; HE staining found that the ankle joint of the model group showed joint cavity narrowing, gnawing-like bone destruction, synovial tissue proliferation and inflammatory cell infiltration. The pathological changes such as destruction of bone tissue, synovial tissue hyperplasia and inflammatory cell infiltration were found in the ankle joint of rats in the model group; compared with the model group, bone tissue hyperplasia and inflammatory infiltration in the ankle joint of rats in the low, medium and high dose groups were significantly improved; TRAP staining observed that compared with the normal group, the model group had the largest number of osteoclasts, and the number of TRAP staining positive osteoclasts was reduced under the intervention of the low, medium and high dose groups; the results of qRT-PCR showed that, compared with the model group. The relative expression of OPG, BMP-2 mRNA was increased in the low, medium and high dose groups (P<0.05), and the relative expression of RANK, RANKL, and TNF-α mRNA was decreased in the low, medium, and high dose groups (P<0.05); the results of WB, compared with the model group, the relative expression of OPG, BMP-2 protein was increased in the low, medium, and high dose groups (P<0.05), and the relative expression of RANK, RANKL , and TNF-α protein relative expression decreased (P<0.05). Conclusion Helleborus thibetanus Franchon may alleviate the inflammatory response in vivo in RA rats by regulating the OPG/RANK/RANKL signaling pathway and exert its role as an anti-rheumatoid arthritis mechanism.
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Biochemical characteristics of type 2 diabetes mellitus and its microvascular complications in animal models

daihaoran, wangyue, wangxu, luijing, wuxiuhong

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Type 2 diabetes mellitus (T2DM) is a metabolic disease characterized by hyperglycemia, which is seriously harmful to human health. Its prevalence rate is increasing throughout the country. Patients with chronic hyperglycemia develop severe microvascular complications such as diabetic nephropathy (DN), diabetic foot (DF), and diabetic retinopathy (DR). Nowadays, T2DM has become a public health problem in the world. How to effectively treat T2DM and prevent microvascular complications has become an important medical issue to be solved urgently. So far, animal models are still the main tools to study the pathophysiology and treatment of T2DM and its microvascular complications, and it is urgent to establish highly consistent animal models. Biochemical indicators are the most direct reflection of the physiological state and body health of animals, and are crucial in evaluating the success of models, often affecting the experimental results. At the same time, biochemical indexes are also important reference basis for disease diagnosis and drug intervention. In this study, the experimental data of the animal model of T2DM and its microvascular complications were summarized, and the biochemical indexes were analyzed, in order to provide theoretical reference for future related experiments.
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Establishment and evaluation of mild to moderate closed traumatic brain injury mouse model

buyuanjing, xudonggang

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【Abstract】Objective To establish a stable mouse model of mild to moderate closed head injury and evaluate its effectiveness. Methods A total of 170 SPF-grade ICR mice were divided into the sham group(n=34) and the TBI group (n=136). The TBI group was subjected to brain injury by impact under conditions A-D, with 34 mice in each subgroup. After modeling, the score of mNss, fatigue rotating test, and Morris water maze test were used to assess the balance and learning ability of the mice; brain water content experiment, Evans blue experiment, HE staining, GFAP immunohistochemistry, and TUNEL immunofluorescence were used to analyze pathological changes in brain tissue. Results Compared with Sham group, mNss score and water maze escape latency were increased to varying degrees in TBI group, while stick stay time was decreased significantly, indicating that severe neurological dysfunction occurred in TBI group. Brain water content and evans blue content in brain tissue of TBI group mice were significantly higher than those in sham group, suggesting that they suffered from blood-brain barrier damage and brain edema. Histopathological examination showed that the neurons in the brain tissue of the TBI group showed obvious degeneration and contraction, the astrocyte proliferation and the proportion of apoptotic cells increased significantly, and the damage degree increased with the increase of the weight of the blow. Comprehensive analysis revealed that a weight of 60 g or 80 g with a height of injury at 20 cm could simulate mild closed head injury in mice; while a weight of 100 g or 120 g could simulate moderate closed head injury in mice. Conclusion A stable mouse model of mild to moderate closed head injury was established based on the principle of free fall, which laid the foundation for studying the mechanism and treatment strategies for mmTBI.
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Based on modified bilateral carotid artery ligation way to establish vascular dementia rat model and to investigate the changing rules of cerebral blood flow and its effects on angiogenesis related pr

chen jie, tang xin, chen pan, xie ziwei, xie haihua, zhang hong, zou yinjie, tan jie

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【Abstract】 Objective Through the modified bilateral carotid artery ligation way to establish VD rat model to observe the changes of cerebral blood flow and the expression of angiogenic proteins. Methods 36 Sprague-Dawley male rats were randomized into sham group（n=18）and model group（n=18）. In the sham group, only the bilateral carotid artery was isolated without ligation, while in the model group, bilateral carotid artery were ligated to establish VD model. Morris water maze behavior test was applied before and 14 days after modeling, the variation of cerebral blood flow was detected by laser speckle contrast imaging. The protein expressions of HIF-1α, VEGF and HO-1 were detected by Western Blotting and the contents of IL-4 and IL-10 were detected by ELISA. Results 14 days after modeling,the escape latency was significantly prolonged and the frequency of crossing the platform had significant decrease for model group as compared with the sham group (P < 0.05). 2 hours,3 days,7 days after modeling, the cerebral blood flow of the model group was significantly lower than that of the sham group (P < 0.05). 14 and 21 days after modeling, there was no significant difference in cerebral blood flow between the sham group and model group (P > 0.05).In the model group, cerebral blood flow decreased to a minimum at 2 hours after modeling (P < 0.05), and then began to recover. The peak of recovery occurred at 3 to 7 days after modeling, and returned to the level before modeling on the 14th day after modeling.At day 21 postoperatively, the expression of HIF-1α, VEGF and HO-1 proteins in hippocampus of the model group were increased remarkably (P < 0.05)and the contents of IL-4 and IL-10 in serum of the model group were significantly rose in comparison with sham group(P < 0.05). Conclusion The variation of cerebral blood flow in VD rats model establish by the modified bilateral carotid artery ligation way showed a certain time rule.At day 21 postoperatively, HIF-1α, VEGF and HO-1 in hippocampus increased significantly, accompanied by increased levels IL-4 and IL-10.
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Preliminary study on the mechanism of infertility in female SD rats with spontaneous dwarfism

long hong, Huo Chunmao, Li Kang, Bao FengYun, Qin TingYang, Zhao YuJia, Zhang ShiBin

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Objective To investigate the causes of infertility and its pathogenic mechanism in female SD rats with spontaneous dwarfism (short stature rat, SSR）.Methods Adult wild-type and SSR female SD rats were used for the study. Vaginal smear was used to observe the changes of motile cycle; ovulation promotion was compared using the simultaneous oestrus supernumerary ovulation method; ovarian and uterine weight and body weight, ovarian and uterine indices were measured; AMH, E2, FSH, LH, FSH/LH levels in serum were measured; transcriptomic sequencing of ovarian tissues will be performed to analyze gene expression differences.Results There were no abnormalities in the estrous cycle of SSR female rats., the body weight of SSR female rats was significantly lower than that of wild type, and their ovarian index and uterine index were significantly higher than that of wild type. The mean number of ovulation was significantly higher in wild-type SD rats than in SSR female infertile rats (p<0.001); serum AMH (p<0.01) and E2 (p<0.05) levels were significantly higher in wild-type SD rats than in SSR female infertile rats, and serum FSH (p<0.01), LH (p<0.01) and FSH/LH (p<0.05) The levels of FSH, LH and FSH/LH (p<0.05) were significantly lower in SSR infertile females than in SSR infertile rats, while PROG was not significant; transcriptome sequencing yielded 250 differentially expressed genes, including 190 up-regulated genes and 60 down-regulated genes. p53 signaling pathway and cytokine-cytokine receptor interaction. The MCC, MNC, EPC and Degree calculations of CytoHubba plug-in were used to screen the top 10 significant nodes, respectively, and the intersection was taken to finally obtain 9 Hub genes, namely Cxcl1, Cxcl2, Il1a, Il1b, Cd80, Mmp13, Mmp8, Fgf3 and Ptgs2.Conclusions Infertility in SSR female rats may be related to decreased ovarian reserve function and poor ovarian response. At the same time, Cxcl1, Cxcl2, Il1a, Il1b, Cd80, Mmp13, Mmp8, Fgf3, and Ptgs2 were screened to be associated with infertility, laying a theoretical foundation for further exploration of infertility mechanisms.
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Research progress on the effects of different ketogenic diets on skeletal muscle performance and fatigue recovery

Li tingting, ZHANG Hui, ZHANG Mingchen, WANG Hongying, ZHANG Pingping, WANG Xiaowen, SUN Zhongguang

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Objective: To summarize the effects of ketogenic diet on sports performance and fatigue recovery of animals and human beings, so as to provide a diet plan for competitive sports and rehabilitation training. Methods: Database PubMed, Web of Science, Embase and CNKI, VIP, WANFANG, CBM were selected. With "(ketogenic diet) and (athletic performance) or (exercise fatigue recovery)"as the retrieval formula, the retrieval period is not limited, and according to the inclusion criteria and exclusion criteria, 42 related literatures were finally included. Results: Ketogenic diet can increase blood ketone, provide energy for skeletal muscle, and play a certain regulatory role in skeletal muscle performance and fatigue recovery. ①Ketogenic diet transforms muscle fiber Ⅱb into Ⅱa through axonal germination and nerve reinnervation, improves the quality and function of mitochondria of fast muscle, and increases histone acetyltransferase to enhance skeletal muscle strength; ②Ketogenic diet uses ketone bodies to provide energy, which can reduce glycolysis and improve the ability of fatty acid oxidation in slow muscles to improve skeletal exercise endurance; ③Ketogenic diet can reduce endoplasmic reticulum stress, oxidative stress and inflammatory reaction of skeletal muscle, protect the body from injury, reduce the consumption of muscle glycogen and the accumulation of lactic acid, relieve fatigue after exercise and promote fatigue recovery. Conclusion: Ketogenic diet has low negative effects on the body, can improve the sports performance and fatigue recovery of animals, plays a maintenance role in humans, and can be used as a diet scheme in competitive sports and rehabilitation training.
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Advances in the Application of Cortisol in Evaluating Stress in Dogs

SUN Ning, FU Jialin, XU Shu, YU Xi, SHUI Yingyi, ZHU Qiwen

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In recent years, cortisol had been used as a biomarker to assess stress in dogs. In order to evaluate the welfare of dogs,we reviewed the cortisol levels and changes in dogs under various stress and explored the influential factors which could relieve stress according to the variation law of cortisol levels ,so?as?to improve the measures to reduce stress in dogs. It is recommended to apply cortisol measurement and behavioral observation comprehensively to evaluate stress in dogs more accurately.
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Mouse and monkey animal models for SARS-CoV-2 infection and pathology

Dong E, ZhangBo, ChenTingwei, Li-Xiaozhuo, Li-Tianqing

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As the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread globally, its genome undergoes ongoing mutations and evolutions, giving rise to different strains such as α, β, γ, δ, etc. Therefore, continuously development of drugs and vaccines targeting different strains becomes pivotal in addressing the COVID-19 pandemic. Constructing animal models of SARS-CoV-2 not only enables studying the pathogenesis of the virus but also stands as a crucial means for evaluating the efficacy of drugs and vaccines related to SARS-CoV-2. However, commonly used animal models such as mice exhibit limited susceptibility to wild-type SARS-CoV-2, underscoring the urgent need for animal models that can be infected with the novel coronavirus and better simulating human pathological and physiological conditions. This review summarizes the animal models used for studying SARS-CoV-2 infection and transmission, as well as their progress in characterizing the viral immunopathology.
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The improvement of the preparation method for type II cardiorenal syndrome rat model

Liu Qian, Wang Xin Ting, Cheng Pei Pei, Rong Jing Feng, Yang Tian Shu, Zhou Hua

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Objective To establish an improved type II cardio-renal syndrome rat model and evaluate it. Methods Twenty male SD rats were randomly divided into the sham group and the model group，with 7 rats in the sham group and 13 rats in the model group. The model group used the method of squeezing the heart under a small animal anesthesia machine to permanently ligate the left anterior descending branch of the coronary artery to cause myocardial infarction. One week later，unilateral nephrectomy (right nephrectomy) was performed. Two groups of rats underwent cardiac echocardiography，pathological staining，and blood and urine tests three weeks after right nephrectomy to verify the establishment of the model. Results Compared with the sham group，the cardiac function detected by echocardiography and the endogenous creatinine clearance rate in the model group rats significantly decreased (P<0.01)，the level of brain natriuretic peptide，blood creatinine, urea nitrogen，and 24-hour urine protein in the model group significantly increased (P<0.01). HE staining revealed disordered myocardial arrangement，atrophy of glomerulus，and infiltration of inflammatory cells in the model group rats. Picric acid Sirius red staining showed a significant increase in myocardial collagen fibers，irregular arrangement of renal tubules，and a large amount of collagen deposition in the model group rats. The positive staining area ratio was significantly increased (P<0.01). Conclusions This improved modeling method can provide a type II cardio-renal syndrome rat model with simple operation，minimal surgical trauma，and low mortality rate. This model simulates the early onset of cardiac and renal function damage and pathological changes in type II CRS，laying the foundation for systematic and in-depth research on the pathogenesis and pharmacological mechanism of type II cardio-renal syndrome.
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Model construction and progress of focused gene therapy for dilated cardiomyopathy

chenyulin, zhaoxinke

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【Abstract】 Dilated cardiomyopathy is one of the main diseases causing heart failure, and there is etiological heterogeneity. Nearly 1/4 of the patients with dilated cardiomyopathy are related to genetics, and ventricular dilation and myocardial systolic dysfunction are the main characteristics of the disease.LMNA mutation is an important cause of hereditary dilated cardiomyopathy, and arrhythmia is an important clinical manifestation of hereditary dilated cardiomyopathy with LMNA mutation.In recent years, the construction and intervention of rodent dilated cardiomyopathy model based on the focused gene therapy of mice with C57/B6 genetic background has been a focus of research, and some important conclusions have been drawn from the construction of large animal models of dogs and pigs. However, large animals, especially non-human primates, are still more close to human models.At present, dilated cardiomyopathy is not involved in the heart disease model of non-human primates. Therefore, this paper reviews the studies on the rodent and large animal models of dilated cardiomyopathy at the genetic level, and also proposes the idea of developing a non-human primate dilated cardiomyopathy model based on the current research.It provides a new idea to study the pathogenesis and clinical treatment in the future.
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Study on the pathogenesis of hyperlipidemia rats under different dietary conditions based on intestinal flora and short-chain fatty acid metabolism

yangzongtong, SUN Tiefeng, lixiaojing, xudongchuan, yuanmin, jinguangqian, wangwenhui

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Objective The purpose of this study was to compare the changes of intestinal flora and the characteristics of short-chain fatty acid metabolism in hyperlipidemic rats induced by two different high-fat diets, and to explore the possible microscopic mechanism of hyperlipidemia from the point of view of host-intestinal flora-metabolism. Methods SPF SD rats were divided into two groups: (1) normal diet group (CG group), fed with high fat diet group (HFD1 group), fed with high fat diet group (HFD1 group), and fed with high fat diet group (HFD2 group). Rats were fed with 80g high fat diet and unlimited maintenance diet.燗fter 8 weeks, the levels of serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) were measured, and the pathological changes of liver tissue and perirenal fat were observed by hematoxylin-eosin (HE) staining.燭he contents of colon were taken for 16s rRNA high-throughput sequencing to observe the changes of the structure and function of intestinal flora and the content of short-chain fatty acids in colon contents. Result Compared with CG group, food intake decreased and body weight increased in HFD1 group and HFD2 group, serum TC, TG and LDL-C increased significantly, liver tissue showed obvious steatosis and perirenal fat showed inflammatory lesions. After high-fat intervention, the relative abundance of intestinal microflora in rats changed significantly and there were gender differences, in which the relative abundance of Lactobacillus decreased significantly, and the structure and function of Lactobacillus decreased significantly, including total short-chain fatty acids, acetic acid, butyric acid and isobutyric acid. Conclusion The two kinds of high-fat diet can cause hyperlipidemia in rats, and the pathogenesis is basically the same, which is related to lipid metabolism and intestinal flora disorder. Daily restriction of a certain amount of high-fat diet can not only reduce the effect of high-fat diet on rat appetite, but also improve the stability of rat hyperlipidemia model preparation.
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Effects of bear bile powder on promotion stage of early hepatocarcinogenesis in SD rats

dongrui, zhangmei, hongzexuan, tangqian, zouxinyu, jiaguiyang, niyuanping, Shibutani Makoto, jinmeilan

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Objective Bear bile powder (BBP), a natural Traditional Chinese medicine (TCM), has antioxidant and anticancer effects. To clarify the modification effect of bear bile powder (BBP) on early period of hepatocarcinoma formation, we conducted an anticancer efficacy experiment using short-term carcinogenesis bioassay models. Method Forty 5-week-old male SD rats were divided randomly into the following 4 groups with 10 animals per group: DEN alone, DEN+PBO, DEN+PBO+BBP-L (200mg/kg BBP), DEN+PBO+BBP-H (400mg/kg BBP). All rats were injected intraperitoneally with 200 mg/kg N-diethylnitrosamine (DEN) and three groups given a diet containing 0.5% piperonyl butoxide (PBO). In addition, the rats of last two groups were orally administered 200 or 400 mg/kg BBP for 8 weeks. Results The relative and absolute liver weights in PBO-treated groups significantly increased compared to DEN alone group. However, the number and area of GST-P+ foci were significantly decreased in only DEN+PBO+BBP-L group compared to DEN+PBO group. In addition, the Ki-67+ cell ratio which are significantly increased by DEN and PBO were significantly decreased after treatment with BBP in both BBP-treated groups. Interestingly, the mRNA levels of Ccne1, Cdkn1b related to cell cycle Caspase 8 and Caspase 9 related to apoptosis were significantly increased only in the BBP-H group. However, these changes were not observed in the BBP-L group. Conclusion These results indicated that BBP has a suppressive effect in the early period of hepatocarcinoma formation, leading to the inhibition of preneoplastic lesions. In addition, these results suggested that the suppression mechanism of BBP is strongly involved in the inhibition of cell proliferation activity and induction of apoptosis. Furthermore, that a high dose of BBP may influence BBP’s inhibitory effect on the preneoplastic lesions.
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Plantar injection of complete Freund's adjuvant induced behavioral and monoamine neurotransmitter changes in pain and depression comorbid model rats

Yang Pu

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Objective To observe the effects of plantar injection of complete Freund's adjuvant (CFA) on pain-depressive behavior and changes in hippocampal monoamine neurotransmitters in rats, with the aim of establishing an animal model of related comorbidity. Methods 16 male, 8-week-old, SPF-grade healthy SD rats were randomly divided into a model group and control group with 8 rats in each group. In the model group, rats were anesthetized and injected with 100 μL of CFA in the left hind paw to induce the comorbid pain and depression model. In the control group, rats were injected with the same volume of saline. Pain thresholds were measured using the von Frey hair and thermal radiation instrument, and depressive-like behaviors were assessed using open field test (OFT), tail suspension test (TST), and forced swim test (FST). Enzyme-linked immunosorbent assay (ELISA) was used to measure the content of 5-hydroxytryptamine (5-HT), dopamine (DA), and norepinephrine (NE) in the rat hippocampal tissue, and histological changes in the hippocampal area were observed by hematoxylin-eosin (HE) staining. Results Compared with the control group, the mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) in the model group were significantly decreased at 3, 7, and 14 days (P < 0.01); the total distance in the OFT was significantly reduced at 7 and 14 days (P < 0.01), and the time spent in the center zone was significantly decreased at 14 days (P < 0.01); the immobility time in the TST was significantly increased at 14 days (P < 0.01), and the immobility time in the FST was significantly increased at 7 and 14 days (P < 0.05, P < 0.01); the content of 5-HT, DA, and NE in the hippocampal tissue of the model group rats was significantly reduced compared with the control group (P < 0.01), and the hippocampal tissue in the model group showed pathological changes, including irregular neuronal shapes, loose and disordered arrangement, increased intercellular space, some unclear cell nuclei, and some neuronal contraction and apoptosis. Conclusion Injection of 100 μL of CFA in the footpad can cause pain hypersensitivity, depressive-like behavior, significant reduction of monoaminergic neurotransmitters in the hippocampus, and histological changes in the hippocampus, effectively simulating the manifestations of comorbid pain and depression, and is an experimental model for studying the pathological mechanisms of comorbid pain and depression.
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The welfare and untilization of experimental animals in tumor research

zhanglingli

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As an important intermediate link between in vitro cell experiments and clinical experiments, animal models are often preferred in the research of tumor mechanisms, prevention,diagnosis, and treatment. The results of animal experiments are directly related to animal welfare, and are important factors affecting the scientific and accurate results of the research. This article summarizes the relevant aspects of experimental animals involved in tumor research, including tumor animal models, animal welfare related to tumor models and humane endpoints.
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Progress in the study of animal models of systemic juvenile idiopathic arthritis and macrophage activation syndrome

Li Feng Ming, Tang Xue Mei

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Systemic juvenile idiopathic arthritis (sJIA) is the most frequent rheumatic disease complicated by macrophage activation syndrome (MAS), which can be life-threatening in children. Current experimental animal models show only some of the characteristics of sJIA and MAS, and there is no perfect animal model that can replicate the whole process of sJIA and MAS. This article reviews the clinical features, advantages and shortcomings of these models, with the aim of providing ideas for exploring more representative animal models of sJIA and MAS.
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Research Progress of Neurotransmitter Detection Technology in Living Brain

Zhu Mingyu, Cui Lili, Chen Huan, Hou Hongwei, HU Qingyuan

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As an endogenous chemical substance, neurotransmitter plays a vital role in maintaining normal life activities of people. Abnormal levels of neurotransmitters can lead to physical, mental and some neurodegenerative diseases. However, the ultra-low concentration, complex chemical properties and release modes of neurotransmitters make their accurate detection in vivo a great challenge.In order to accurately monitor neurotransmitters in the brain and accurately understand the release kinetics of neurotransmitters, several commonly used methods for detecting neurotransmitters in vivo in the past five years and their research progress were reviewed.The basic principle and applicability of microdialysis, electrochemical sensor and fluorescence sensor are introduced in detail.
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Research progress on animal models of acute exacerbation of various respiratory diseases induced by compound factors

Qiu Zhiguang, SHAO Xuejie, LU Ruilong, TAN Yange, REN Zhouxin

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Based on the compound factors of clinical practice, some progress has been made in acute exacerbation models of respiratory diseases by "two-hit" method. In this review, the current research on acute exacerbation models of pulmonary fibrosis, chronic obstructive pulmonary disease and bronchial asthma constructed by compound factors is summarized. Furthermore, the characteristics and scope of application of each model are compared and analyzed in animal strain selection, model preparation methods and major histopathological changes, providing reference for researchers to further improve and perfect the model or rationally select the animal models.
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Effect of miR-185-5p on proliferation, invasion and migration of mouse breast cancer PY8119 cells

Liu Lu, Zhao Yanqiao, Wang Xiaona, Wang Kun, Yin Chonggao, Liu Qinghua, Li Hongli

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Object To investigate the effect of miR-185-5p on the proliferation, invasion and migration of mouse breast cancer PY8119 cells. Methods Mirna: miR-185-5p with significantly down-regulated expression in breast cancer was obtained by previous research group. The sequence of miR-185-5p gene from human and mouse was consistent by NCBI. The effects of overexpression and knockdown of miR-185-5p on the proliferation of PY8119 cells were detected by EdU proliferation experiment. The effects of overexpression and knockdown of miR-185-5p on the invasion ability of PY8119 cells were detected by Transwell invasion experiment. The effects of overexpression and knockdown of miR-185-5p on the migration ability of PY8119 cells were detected by scratch healing experiment. The effects of overexpression and knockdown of miR-185-5p on the apoptosis of PY8119 cells were detected by flow cytometry. The effect of overexpression of miR-185-5p on tumor proliferation in vivo was detected by C57BL/6 mouse subcutaneous tumor model. Lung metastasis of mice overexpressing miR-185-5p was observed by tail vein injection. Results The results of EdU proliferation experiment, Transwell invasion experiment, scratch healing experiment and flow cytometry showed that knocking down miR-185-5p significantly enhanced the proliferation, invasion and migration of PY8119 cells and inhibited apoptosis. Overexpression of miR-185-5p inhibited the proliferation, invasion and migration of PY8119 cells and promoted apoptosis. In vivo tumorigenesis experiment of C57BL/6 mice showed that overexpression of miR-185-5p slowed down the tumor growth rate in C57BL/6 mice. Lung metastasis experiment showed that overexpression of miR-185-5p inhibited the lung metastasis of C57BL/6 mice. Conclusion miR-185-5p can inhibit the proliferation, invasion and migration of mouse breast cancer PY8119 cells in vitro and in vivo as a tumor suppressor gene.
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Research hotspots and trends of Alzheimer's disease animal experiments: CiteSpace bibliometric analysis based on WOS database

YANG Jia-hui, LIN Yin, CHEN Lai, luoxiaoquan

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Objective Alzheimer's disease (AD) brings a huge burden to patients and society. As an important means of studying AD, animal experiments provide a lot of scientific basis for clinical and scientific researchers. This paper adopts the method of bibliometric analysis to reveal the research status, hotspots and trends in the field of AD animal experiments. Methods We obtained publications in the field of AD animal experiments from 2018 to 2022 from the Web of Science Core Collection (WoSCC) database. We use the analysis software CiteSpace 6.2.R2 (Advanced) to analyze the general information and keywords of publications in this field. Results After analyzing 4015 articles included in this study from 2018 to 2022, it was found that the annual publication volume of papers has gradually increased. In terms of the number of articles, the United States, the University of California System, Saito T, and the International Journal of Molecular Science are the countries, institutions, authors, and journals with the largest number of articles. "Alzheimer's disease" had the highest frequency of occurrence, and "anxiety" had the highest central degree. Conclusion This field focuses on the study of pathogenesis and treatment methods, and the plasticity of entorhinal cortex, neurons and synapses is a hotspot of current research. Future research trends may mainly focus on Aβ plaque formation, tau hyperphosphorylation, neurofibrillary tangle accumulation, glial inflammation, and ultimately loss of proper neuroplasticity. This article visually analyzes the hotspots and trends of AD animal experiments to help researchers understand the latest situation.
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Study on the expression and methylation level of hippocampal Notch signaling pathway genes in perimenopausal depression rats

shen jian ying, liang wen na, chen hui fang, zhang ling yuan, zhuang yuan ying, chen xiao yang, chen ya ru, xiao qiong qiong, yang min, gong lin, min li

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[Abstract] Objective To investigate the effect of perimenopausal depression on the expression of key genes of Notch signaling pathway and DNA methylation of its promoter in rat hippocampus from the perspective of DNA methylation of epigenetics. Methods 12-month-old female SD rats were randomly divided into control group and model group. Open field tests, sugar consumption tests, and forced swimming tests were used for behavioral evaluation. Real-time PCR was used to detect the expression of key genes of Notch signaling pathway in the hippocampal dentate gyrus of different groups. Bisulfite sequencing (BSP) was used to analyze methylation sites and methylation levels in the promoter region of key genes of the Notch signaling pathway. Results In perimenopausal depression rats, the horizontal and vertical scores of open field test and the rate of sugar water consumption decreased significantly; the expression of the key genes Jagged1, Notch1 and Hes5 on Notch signaling pathway was significantly down regulated in the hippocampus of perimenopausal depression rats (P＜0.01 or P＜0.05) ; among them, some methylation sites and methylation rates of Hes5 gene increased significantly, while the methylation sites and methylation rates of Jagged1 and Notch1 genes were not statistically significant (P>0.05) . Conclusion The expression of key genes of Notch1 signaling pathway in the hippocampal dentate gyrus of perimenopausal depression rats is significantly down regulated, and the down-regulation of Hes5 gene may be related to the increase of promoter DNA methylation level.
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Establishment and evaluation of rats model of rheumatoid arthritis with depression

liyuanyuan, zhangbitao, fanpeijian, cuizilong, wangshaoxian

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Objective To prepare and evaluate the animal model of rheumatoid arthritis with depression (RAD). Methods Thirty-two rats were randomly divided into normal group (N group), depression group (D group), arthritis group (RA group) and model group (RAD group), with 8 rats in each group. The N group was fed normally, while the D group was fed solitary rearing for 1 week and chronic unpredictable mild stress (CUMS) for 3 weeks, RA group was induced by collagen-induced arthritis (CIA) to establish the arthritis model. In RAD group, RAD animal model was established on the basis of CIA animal model, meanwhile, adding 1 week solitary rearing and 3 weeks CUMS. Toe volume, arthrits index (AI), Synovial and ankle joint pathology, serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were used to evaluate whether the rat model of rheumatoid arthritis was successfully established; the depression status of rats was evaluated by measuring body weight, food intake, sugar water preference rate, open field behavior (total number of motion cells, number of standing uprights), serum levels of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), corticosterone (CORT), and pathological changes of hippocampus. The RAD animal model was successfully prepared if depression and rheumatoid arthritis were both tested in the rats. Results Compared with the N group, toe volume, AI score and the levels of IL-1β, IL-6 in serum of RA group and RAD group were significantly increased (P＜0.05 or P＜0.01), the joint space widened, synovial hyperplasia was obvious and extended to the joint cavity, and a large number of inflammatory cells were seen; in D group and RAD group, body weight, food intake, total number of motion cells, number of standing uprights and sugar water preference rate of rats were decreased to different degrees, and the levels of CRH, ACTH, CORT in serum were increased in different degrees (P＜0.05 or P＜0.01), the neuron cells arranged loosely, gap increased, the number decreased, accompanied by some nucleus pyknosis, dissolution, rupture, especially in the model group. Conclusion CIA combined with solitary rearing and CUMS can better simulate the changes in the external manifestations and internal indicators of RAD model animals, which can provide reference for the study of RAD disease animal models.
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Quantitative inhalation method to established the model of chronic obstructive pulmonary disease in rats

liuxuewu

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Objective To compare the differences of chronic obstructive pulmonary disease (COPD) models induced by smoke inhalation through nose-mouth plus LPS or smoke exposure through wholebody plus LPS in rats, providing a new model for COPD model construction. Methods 90 male SD rats were randomly divided into normal control group, wholebody exposure group and nose-mouth inhalation group, with 30 rats/group. The wholebody exposure group were exposed in a homemade smoke box where smoke contacted with whole body of rats , and the smoke nose-mouth inhalation group were inhaled with somke via nose-mouth only in a "quantitative smoking device". Animals in both groups were exposed to smoke once a day for 60mins/time for 8 weeks, and LPS (1mg/kg) was injected through the trachea on day 1, 7, 15 and 21, respectively, to induce the COPD model. The quality control of smoke generated by quantitative smoking devices included the verification of the stability and uniformity of the concentration of smoke particles and the size distribution of smoke particles. At 4, 6 and 8 weeks of modeling period, pulmonary function examination, the content of inflammatory factor interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) in alveolar lavage fluid and histopathological examination were performed to compare the differences between the two modeling methods. Results Quantitative smoking devices could produce smoke with stable concentrations of 1.1mg/L (counted as total particles) and 0.1mg/L (counted as nicotine), respectively, with a median mass particle size of 0.693μm (in nicotine) and a GSD of 1.463. Compared with the whole body exposure group, the indexes of pulmonary function FEV0.2/FVC and pulmonary compliance (Cdyn) in the nose-mouth inhalation group decreased more significantly, and the airway resistance (Penh) increased more significantly. The levels of IL-6 and TNF-α in the alveolar lavage fluid of rats in the nose-mouth inhalation group were significantly increased at 6 weeks after modeling, while those indexs in the wholebody exposure group were increased at 8 weeks after modeling. The lesion severity of bronchial inflammation after modeling was similar between the oral-nasal inhalation group and the wholebody exposure group, but the lesion severity of emphysema was more serious in the oral-nasal inhalation group (the time when appeared statistic difference: oral-nasal inhalation vs. wholebody exposure = 6 weeks of modeling vs. 8 weeks of modeling). The mean linear intercept (MLI) in the oral-nasal inhalation group increased significantly at 4-8 weeks of modeling, and the mean alveolar numbers (MAN) decreased significantly at 6-8 weeks of modeling. MLI increased significantly and MAN decreased significantly in the whole body exposure group after 8 weeks of modeling. In the oral-nasal inhalation group, significant abnormal changes were observed in pulmonary function indexes (FEV0.2/FVC, Cdyn, Penh), cytokine levels in Bronchoalveolar lavage fluid (IL-6, TNF-α), and alveolar histopathological changes (bronchial severity and emphysema pathological score, MLI, MAN) after modeling. However, the coefficient of variation (CV%) of each index was significantly lower than that of the whole body exposure group. Conclusion LPS (1mg/kg) endotracheal drip combined with smoke wholebody exposured or via oral-nasal inhalation both could establish a typical rat COPD model. Rats Inhalated smoke via oral-nasal could shorten the modeling period. The model could be completed after 6 weeks of continuous modeling, presenting typical symptoms of chronic obstructive pulmonary disease (pulmonary ventilation dysfunction, broncho-lung chronic inflammatory infiltration, accompanied by emphysema), and the difference between individual model animals were smaller (vs smoke exposure, CV% values were smaller).
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Research progress of animal metacognition

WANG WANNING, JIA NING

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It is widely believed that metacognition is a special cognitive capacity that only humans possess. A contentious debate in the field of comparative psychology is whether animals have metacognition. The exploration of this question has become an important way to uncover the phylogenetic roots of metacognition and to map the evolutionary history of higher cognitive functions in humans. This paper reviewed the main animal metacognition studies in the past 25 years. We summarize three common research paradigms and corresponding measures: the uncertainty monitoring paradigm, the information seeking paradigm, and the confidence judgment paradigm. Two opposing explanatory tendencies and their rationales are outlined in the field of animal metacognition research: the associative model and the metacognitive model. In response to the debate between the associative model and the metacognitive model, two future research orientations are proposed to improve the research paradigm to test the metacognitive model and the innovation theory to integrate the model.
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Data mining based animal model for dilated cardiomyopathy

gejunxi

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Objective To review the animal species, modelling methods and evaluation indexes of dilated cardiomyopathy (DCM) animal models at home and abroad, to provide suggestions for improving the success rate of modelling and establishing DCM animal models with Chinese medicine characteristics in the future, and to provide reference for the establishment of future DCM animal models. Methods The literature on animal models of dilated cardiomyopathy in the past 10 years was searched, and relevant reports were collected and summarized. Relevant parameters of experimental animals (species, sex, body weight, etc.), modelling methods, modelling cycles, positive control drug species and cycles, and model evaluation indexes were summarized and analysed. Results A total of 128 papers were included. Animal models of dilated cardiomyopathy were mostly prepared in SD rats and Wistar rats using the anthracycline adriamycin, with a cycle of 30 d
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To investigate the effect of TMAO on lipid metabolism in spleen deficiency hyperlipemia rats and the intervention effect of Xiangsha Liujunzi decoction

lengxue, 李阳

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Objective To investigate the effect of TMAO, a metabolite of intestinal flora, on hepatic lipid metabolism in rats with splenic deficiency and hyperlipidemia, and to further explore the possible mechanism of Xiangsha Liujunzi Decoction in the intervention of hepatic lipid metabolism disorder. Methods SD rats were divided into: (1) Blank control group (group C), (2) blank control group DMB (group C D), DMB is TMAO inhibitor, (3) spleen deficiency hyperlipidemia group (PG group), (4) spleen deficiency hyperlipidemia DMB group (PG D), (5) Spleen deficiency hyperlipidemia Xiangsha Liujunzi Decoction group (PG XS group), Except group C and group C D, the other groups were constructed to establish spleen deficiency hyperlipidemia model (12 weeks of modeling) by combining excessive fatigue and high-fat diet. After model establishment, group C D and PG D were given 1%DMB in drinking water every day, and group PG XS was given Xiangsha Liujunzi decoction (11.34 g crude drug /kg/ day) every day. The other groups were given the same amount of normal saline. The blood lipid level of each group was detected by automatic biochemical method after 4 weeks of intragastric administration. The morphological changes of liver were observed by HE staining. The lipid deposition in the liver of each group was observed by oil red O method. Liver FFA, TG and TC were detected by ELISA. The plasma TMAO content was detected by LC-MS. The relative expression levels of PERK, FOXO1, SREBP-2, ABCA1 and miR-33 mRNA in liver were detected by qRT-PCR. The contents of SREBP-2 and ABCA1 gene protein in liver were detected by WB. Results (1) The content of TC, TG and LDL-C in serum of PG group was significantly higher than that in C group, and the content of HDL-C was significantly lower than that in C group; The contents of FFA, TG and TC in liver tissue of PG rats were significantly increased compared with those of C group. Compared with group C, lipid deposition in liver was aggravated and fat vacuoles were increased significantly. There was no difference between group C D and group C in the above indexes. Compared with the PG group, PG D and PG XS groups could significantly reduce the contents of TC, TG and LDL-C in serum, increase the content of HDL-C, reduce the contents of FFA, TG and TC in liver tissue, alleviate the phenomenon of lipid deposition in liver tissue and reduce liver vacuoles. There was no significant difference between PG D group and PG XS group. (2) Compared with group C, the plasma TMAO content of PG group was significantly increased, the liver PERK, FOXO1 and miR-33a mRNA expressions were significantly increased, and the liver SREBP-2 and ABCA1 mRNA and protein expressions were significantly decreased; There was no difference between group C D and group C in the above indexes. Compared with the PG group, PG D and PG XS groups could significantly reduce the plasma TMAO content, decrease the liver PERK, FOXO1 and miR-33a mRNA expressions, and increase the liver SREBP-2 and ABCA1 mRNA and protein expressions. There was no significant difference between PG D group and PG XS group. Conclusion TMAO may regulate the SREBP-2/miR-33a/ABCA1 signaling pathway through PERK/FOXO1 axis to cause liver lipid metabolism disorders in rats, and Xiangsha Liujunzi Decoction may inhibit liver lipid metabolism disorders by inhibiting TMAO content.
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Study on the effect of Huanglian Jiedu Decoction on bile acid metabolism in SD rats based on serum metabolomics

yangguangyong, duhaiyang, zhanggengxin, sugang, tuxiaohua, heguangzhi

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Objective To study the effect of Huanglian Jiedu Decoction(HLJD) on the physiological function of healthy SD rats through serum non-targeted metabolomics. Methods After 7 days of adaptive feeding, 24 rats were randomly divided into blank group (group N) and high-dose HLJD group (group H), medium-dose HLJD group (group M), and low-dose HLJD group (group L) with six rats per group. Group N was gavaged with physiological saline, and H, M, and L groups were gavaged with 6.250, 3.125, and 1.560 g/kg HLJD, respectively. Rats were gavaged with 1 mL twice per day for 21 d. Before collecting serum samples, fasting and water deprivation were performed for 12 h. After anesthesia, blood was collected from the abdominal aorta, and serum was separated. Changes in metabolic products in the serum were detected by Gas Chromatography Time-of-Flight Mass Spectrometry(GC-TOF-MS). Multivariate analysis by Orthogonal Projections to Latent Structures Discriminant Analysis(OPLS-DA) and univariate statistical methods were used to screen differential metabolites, and then metabolic pathway analysis was conducted using the Metabo Analyst platform. Results Compared with the group N, metabolic small molecules were significantly altered in the serum of group H, group M, and group L, which mainly included cholesterol, cholic acid, ribose, paclitaxel, cortisol, oleic acid, succinic acid, linolenic acid, acetic acid, and acetone alcohol (P < 0.05). Correlation analysis showed that HLJD significantly affected the metabolism of primary bile acids, alanine, aspartic acid, and glutamate in α-linolenic acid and glycerol phospholipid metabolism, and its in vivo effects may be related to the intake dose, providing a reference to further reveal the mechanism of HLJD in promoting bile acid metabolism. Conclusions HLJD promotes the production of primary bile acids in vivo and enhances the effects of the bile acid metabolic pathway.
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The model of ulcerative colitis with spleen deficiency and dampness syndrome and the changes of T lymphocytes

qiuping xiao, zhong youbao, yu songren, li shangshang, luo xiaoquan, chen liling, liu xuan

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Objective A mouse model of ulcerative colitis with spleen deficiency and dampness syndrome was established, and the role of T lymphocytes in this model was studied. Methods C57BL/6 mice were randomly divided equally into Control, FXY, 3% DSS, FXY+3% DSS and FXY+2% DSS groups. A mouse model of ulcerative colitis with spleen deficiency and dampness syndrome was established by DSS and senna leaf. At the end of moulding, HE staining was performed to observe structural changes in the colon tissue. Elisa assay was performed to detect the concentrations of inflammatory cytokines in the colon tissues. Flow cytometry was used to detect the levels of CD4/CD8 T lymphocytes, Th1/Th2, Th17/Treg cells. Results In the FXY group, the clinical symptoms of mice with spleen deficiency and dampness were mainly diarrhea, and mice with 3% DSS colitis mostly showed blood in stool, while mice with spleen deficiency and dampness colitis in the FXY+3%DSS and FXY+2%DSS groups mostly showed diarrhea and blood in stool, but the survival rate of mice in the FXY+3%DSS group was as low as 50%. Compared with the Control group, body weight, colon length and concentrations of anti-inflammatory cytokines IL-10 and TGF-β1 were significantly lower in the 3%DSS, FXY+3%DSS and FXY+2%DSS groups, while concentrations of DAI, colon weight, colon weight index, colon weight/colon length, pro-inflammatory cytokines TNF-ɑ, IL-1β and IL-6 were significantly increased, and significant ulcer formation and inflammatory cell infiltration were observed under light microscopy. Compared with the Control group, the percentages of CD4, Th1 and Th17 cells in the mesenteric lymph nodes of FXY+3% DSS and FXY+2% DSS groups were significantly increased, while CD8, Th2 and Treg cells were significantly decreased. Conclusion The combination of Senna and 2% DSS could successfully construct a model of mice with ulcerative colitis with spleen deficiency and dampness evidence, showing typical CD4/CD8, Th1/Th2 and Th17/Treg cell imbalance characteristics.
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Progress in the application of the zebrafish model in the study of neurodegenerative diseases

Liu Yanying

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【Abstract】 With the process of the aging society, the incidence rate of neurodegenerative diseases is increasing year by year. Zebrafish have many advantages that other model organisms do not have, such as unique growth and development mode, low price and easy reproduction, and highly similar to human genes. In the past decades, zebrafish as a model organism has been widely used in various research fields of medicine and life science. This article reviews the application and progress of zebrafish in the research of several common neurodegenerative diseases in recent years, including Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinal muscular atrophy, hereditary spastic paraplegia, and other diseases related to the nervous system. This review article analyzes and summarizes the advantages and disadvantages of zebrafish as an ideal model organism. In future research, especially in the mechanism research of neurodegenerative diseases and large-scale screening of therapeutic drugs, zebrafish has considerable application prospects.
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Research progress on animal model construction and evaluation of different TCM syndromes of gastric ulcer

Wang Wen, Hou Yujun, Wang Lu, Yan Xiangyun, Li Yanqiu, Sun Luqiang, Chen Shuai, Shi Yinzhou, Zheng Qianhua, Li Ying

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Gastric ulcer is a common digestive system disease. Traditional Chinese medicine (TCM) has a significant effect in treatment of this disease. However, its treatment mechanism is unclear. Establishing a scientific and objective animal model and model evaluation system is of great significance to promote development of TCM in this field. This article organizes the literature related to treatment of gastric ulcer by TCM in the past 10 years and summarizes methods for establishing and evaluating animal models of TCM syndromes relating to gastric ulcer. The methods were reviewed for liver and stomach disharmony syndrome, spleen and stomach dampness-heat syndrome, blood stasis in stomach syndrome, deficiency cold of spleen and stomach syndrome, stomach Yin deficiency syndrome, and liver depression and spleen deficiency syndrome. The current animal models of gastric ulcer had the following problems: the typing of TCM syndromes was not unified, the methods and syndrome types of the models were limited, the standards of model establishment methods were different, the combination of disease and syndrome of models was not close enough, and the model evaluation system was not standardized. Suggestions for model improvement are proposed to provide a reference for research in this field.
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Analysis of pathological changes of knee joint and intestinal tissue in cynotonic spondylitis monkeys

huangyuye, caiyanzhen, caichunmei, zhuhepeng, dinghuangguan, xiaowende, chenjun, luli

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Objective: Ankylosing spondylitis (AS) is an immune-mediated chronic disease involving axial joints, peripheral joints and intestines, but the etiology of peripheral knee and intestine has been unknown. This study aimed to study the main pathological changes and mechanisms of knee joint and intestine in cynomolgus monkeys with AS. Methods: Through histomorphological analysis and immunohistochemical experiments, the key characteristics of peripheral knee joint and intestinal tissues of AS were studied and the possible pathogenesis was preliminarily analyzed. Results: The pathological characteristics of peripheral knee joints in AS cynomolgus monkeys mainly showed cartilage erosion on the joint surface, exposed subchondral bone, and jagged joint surface in the early stage. In the late stage, it is mainly manifested as ectopic hyperplasia of cartilage superficial hypertrophy of chondrocytes, and osteophytes are formed through chondroplaginous osteogenesis and fibroogenesis, and cartilage is basically lost. The expression of MMP-3 in cartilage and blood vessels is upregulated, causing cartilage destruction and stimulating angiogenesis. AS cynomolgus monkeys have severe atrophy of small intestinal villi and obvious crypt hyperplasia, and a large number of γδ T cells can be seen in the mucosal intestinal glands of the jejunum and ileum. Conclusion: In this study, the pathological analysis of knee joint and intestine of AS cynomolgus monkey obtained the key characteristics of knee joint and intestinal tissue of this spontaneous model, and proposed a possible pathogenesis. This paper provides new insights into the potential link between autoimmunity in bone tissue and intestinal tissue lesions in AS.
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Langendorff Cardiac Perfusion Technique (for Myocardial Ischemia Reperfusion) applications and developments

Linan, Zhang Chen, Sun yanjun, Gao Shengwei, Li Duojing, Wang xinrui, Wang Baohe

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The Langendorff ex vivo cardiac perfusion model technique is reproducible, easy to administer and has low technical requirements, and is often used to study myocardial ischemia-reperfusion injury diseases. This article reviews the steps of this technique, selection of animals, perfusion patterns, selection of perfusate, indices of detection, ischemic modalities, and selection of ischemia and perfusion times.
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Study on the antidiarrheal effect of new "Ershen pills", a composition of new sliced nutmeg Koji

Fan Mengmeng, Li Kai

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Objective To evaluate the antidiarrheal effect of new ershen pills composed of nutmeg koji instead of bran stewed nutmeg.Method Kunming mice were randomly divided into normal group, model group, Ershen pills Ⅰ (salt psoralen + bran stewed nutmeg), Ershen pills Ⅱ (salt psoralen + nutmeg koji), Ershen pills Ⅲ (salt psoralen + nutmeg raw product), salt psoralen group, and nutmeg koji group. The combined modeling method of "hydrocortisone + senna leaf" was used to establish the diarrhea mouse model with spleen-kidney Yang deficiency. The general signs and pathological changes of each organ were observed. The levels of various organs index, small intestine propulsion rate, gastric residual rate, serum motilin (MTL), gastrin (GAS), adrenal ketone (CORT), thyroid stimulating hormone (TSH), testosterone (T), tumor necrosis factor-α (TNF-α) and interleukin 1β (IL-1β) were detected. 16S rDNA sequencing and information analysis were conducted for fecal microorganisms. Results After modeling, compared with normal group, the weight and activity of mice in model group were reduced, small intestine propulsion rate was significantly increased, gastric residual rate and organ index were significantly decreased, serum GAS, CORT, TSH and T levels were significantly decreased, MTL, TNF-α and IL-β levels were significantly increased, and intestinal flora species diversity was decreased. After administration, the above indexes and symptoms were improved in different degrees in each administration group, and Ershen Pills group Ⅱ was better than Ershen Pills group Ⅰ, Ershen Pills group Ⅲ, salt-psoralea and myristoqu groups. Conclusion The combined use of nutmeg koji and salt psoralen has a remarkable effect on the diarrhea of spleen and kidney Yang deficiency. The fermented nutmeg can reduce its toxic risk and enhance its effect of warming spleen and preventing diarrhea, which laid a foundation for the development of nutmeg koji.
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Characteristics and application analysis of ovarian cancer animal model

yi chu, zhan sha, ma xin yi, wu yang jie, bu jun yi, ma min, yan xian xin

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【Abstract】Objective To study the characteristics and application status of animal models of ovarian cancer and provide guidance for standardized preparation of ovarian cancer models. Methods Using “ovarian cancer” and “animal model” as the main topics in CNKI, Wanfang, VIP, and PubMed databases, the databases were searched for animal experimentation literature on ovarian cancer from establishment of the database to March 1, 2023. A total of 1428 relevant articles were collected. From the experimental animal species, age, experimental modeling methods, detection indicators, and other aspects of the summary, we established a database for systematic analysis. Results A total of 178 studies were obtained after screening. Female BALB/c mice were selected to establish the animal model of ovarian cancer. Most choices were 4 ~ 6 and 6 ~ 8 weeks of age. The most common modeling method was ectopic transplantation, which was mostly subcutaneous injection in the axilla. In terms of modeling methods of ovarian cancer in animals, transplantable models were the most used, and ectopic transplantation was more often employed than orthotopic transplantation. The ovarian cancer cell lines used were mostly human. Most detection indexes were the appearance of tumor tissue, tumor pathology, and immunohistochemistry. Conclusion Animal models of ovarian cancer are widely used in ovarian cancer research, but standard preparation and evaluations are lacking. We analyzed the application status of animal models of ovarian cancer in detail through literature collation and data classification analysis to provide a reference to establish standardized animal models of ovarian cancer. BALB/c mice are the most selected, and human ovarian cancer cell line SKOV3 at 1 × 107 cells/ml is commonly injected into the subcutaneous axillary area for 10 days. A large number of models can be obtained in a short time with a high tumor formation rate and small individual differences, which provide a reference to study ovarian cancer.
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Model establishment and evaluation of rats with chronic obstructive pulmonary disease complicated with chronic kidney disease

Fanzhengyuan, Li Ya, LI Suyun, LI Gaofeng, LI Jingmei

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Objective To establish and evaluate a model of chronic obstructive pulmonary disease (COPD) combined with chronic kidney disease (CKD) in rats. Methods Forty SPF grade SD rats were randomly divided into control, COPD, CKD, and COPD combined with CKD model (COPD+CKD) groups, with 10 rats in each group. The COPD rat model was prepared by cigarette smoke exposure combined with the bacterial drip method, the CKD rat model was established by adenine induction, and the COPD combined with CKD rat model was prepared by both methods. Results After successful modeling, lung function indexes incluidng forced vital capacity (FVC), forced expiratory volume in 0.1s (FEV0.1), and FEV0.1/FVC were significantly reduced in the COPD+CKD group (P < 0.01), and lung histopathology showed emphysematous changes with alveolar wall fracture and fusion as well as inflammatory cell infiltration. Serum Cr, BUN, and 24 h urine protein were significantly increased (P < 0.01). Renal histopathology showed glomerular mesentery proliferation, basement membrane thickening, tubular dilatation, and interstitial fibrosis. The ultrastructure showed that glomerular capillary loops were partially closed, foot processes were fused, renal tubule mitochondria were fused and disintegrated, and lysosome was increased. Serum IL-6, IL-13, IL-1β, and TGF-β1 levels were significantly increased (P < 0.01) and were significantly higher than those in single model groups (P < 0.01). Conclusions Cigarette smoke exposure combined with bacterial infection and 2.5% adenine induction successfully establishes a model of COPD complicated with CKD in rats, and the inflammatory response might play a major role in the process of COPD complicated with CKD.
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Research progress of functional dyspepsia animal model based on the combination of disease and syndrome

liuhao, wangxinai, liwenjing, wuyaxuan, wangchenqi, xiaohongling

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Functional dyspepsia (FD) is a non-organic disease mainly due to regional dysfunction of stomach and duodenum. In traditional Chinese medicine(TCM), it is classified as "epigastric fullness" and "epigastric pain", and divided into the syndrome types of spleen deficiency and qi stagnation, spleen-stomach deficiency-cold(weak), spleen-stomach damp-heat, liver-stomach disharmony, and cold- heat complicated. TCM has a significant effect and high acceptance in the treatment of FD, and the animal model with the combination of disease and syndrome is the basis and prerequisite for relevant research. Therefore, this paper organized and summarized the existing research on animal models of FD combined with disease and syndrome, explored the reasons for the selection of specific modeling methods and evaluation indicators, and proposed the shortcomings of the current research, in order to provide support for the future research on animal models of FD combined with disease and syndrome.
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Collagen peptide combined with sodium alginate improved intrauterine adhesions in mice by down-regulating the expression of NLRP3 inflammasome

chengyiyi, zhaoweiwei, song zhenfeng, tongfuwen, liugang, zhanghua, shileilei, guozhikun, wangxianwei, panying, sunyongkun

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【Abstract】Objective To investigate the repair effect and mechanism of collagen peptides (CP) combined with sodium alginate (SA) on endometrial injury. Methods Forty-eight SPF female C57BL/6N mice were randomly divided into sham group (16 mice), model group (16 mice) and CP SA group (16 mice). In the model group, 95% ethanol was injected into the uterine cavity through the cervix to induce endometrial injury, and the intrauterine adhesion model was established. The sham operation group was injected with 0.9% normal saline into the uterine cavity. The treatment group was injected with the mixture of CP SA after the injection of 95% ethanol into the uterine cavity. After 7 days of modeling, 8 mice in each group were selected to collect samples for each test. HE staining was used to observe the pathological changes of mouse uterine tissue. Modified Masson tri-color staining was used to observe endometrial fibrosis. Western blot was used to detect the protein expression levels of inflammatory factors NLRP3, ASC, Caspase-1, IL-1β and IL-18 in uterine tissues. ELISA was used to detect the expression of IL-1β and IL-18 in serum. The remaining 8 mice in each group were cohoused according to the male-female ratio of 1:2, and the number of pregnant mice in each group was recorded. Pregnant mice were euthanized in the 14th day of pregnancy, and the number of embryos was recorded. Results (1) Compared with the sham operation group, the model group showed thinner endometrial thickness, increased endometrial fibrosis, and inflammatory cell infiltration. In addition, the protein expression levels of inflammatory factors NLRP3, ASC, Caspase-1, IL-1β and IL-18 in uterine tissue and the levels of IL-1β and IL-18 in serum in the model group were significantly lower than those in the sham operation group (P<0.05). (2) Compared with the model group, the treatment group showed that the degree of endometrial injury, endometrial fibrosis and inflammatory cell infiltration were reduced. In addition, the protein expression levels of inflammatory factors NLRP3, ASC, Caspase-1, IL-1β and IL-18 in uterine tissue and the levels of IL-1β and IL-18 in serum in the model group were significantly decreased than those in the sham operation group (P< 0.05). (3) Compared with the sham group, the reproductive ability of the model group decreased (P<0.05). Compared with the model group, the reproductive ability of the treatment group was significantly improved(P<0.05). Conclusions The combined application of CP and SA can effectively improve the pathological changes of uterine tissue, reduce endometrial injury, inflammatory response and fibrosis, and improve the reproductive ability of mice. The mechanism may be related to the down-regulation of NLRP3 inflammasome expression, which provides a reliable experimental basis for the combined application of CP and SA in the treatment of intrauterine adhesion.
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Research progress of bronchoalveolar lavage in mice

Wu Zhihao, Yang Luyin, Ren Wei, Zhou Yanan, Wang Hong, LeiYun, Yu Hong, Yang Sijin

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Bronchoalveolar lavage is an important technique for studying respiratory diseases and has been widely used in disease research. Currently, the operation procedure of bronchoalveolar lavage in clinical patients has been gradually standardized, while mice, as one of the important model animals for studying lung diseases, still need a standard collection and testing procedure for bronchoalveolar lavage fluid for mice. An unstandardized procedure can hinder the promotion and application of this technique in research, and also affect the accuracy and reliability of experimental results. This paper presents a systematic summary of bronchoalveolar lavage methods for mice in domestic and international studies, with the aim of providing reference and guidance for researchers to apply and establish a standardized lavage procedure in the future.
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Advances in Animal model for studying albumin-related drugs

Ziyin, Ning Li, Tang Chengcheng, Zhou Xiaoqing

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The main challenges of therapeutic peptides or proteins is their short plasma half-life. The drugs combining these peptides or proteins were defined as albumin-related drugs. Such combination can effectively prolong the plasma half-life. Animal model is an important and indispensable tool for studying drug pharmacokinetics. This review summarizes two main animal models; rodents (wild-type mice, gene-modified mouse models, rat models) and non-rodents (monkeys), and their development and application in albumin-related drugs. This review can provide a reference for further studies on albumin-related drugs.
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Research progress on the establishment of mouse atherosclerosis model by different intervention factors

baixufeng, cao qingyu, 胡, wang shuwen, liu yali

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Atherosclerosis (AS) is a potential risk factor for common cardiovascular and cerebrovascular diseases. Prevention and treatment of AS and research and development of anti-AS drugs have been a focus in the field of medicine. Selection of the ideal AS animal model is important to study prevention and control measures of AS and to research and develop traditional Chinese medicine for AS. The ideal animal model of AS should have similar pathogenesis to human AS, consistent pathological and biochemical reactions, high repeatability, simple operation, and easy adoption. Mice have the characteristics of strong reproduction, high heritability, good plasticity, and fast modeling of AS, and they have become ideal animals for AS research. In this article, the replication methods of the mouse AS model established by various intervention factors were compared and analyzed to provide a reference and ideas for prevention and treatment of AS and research and development of traditional Chinese medicine for AS.
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dong

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Application and development of germ-free animals for the gut microbiome research

Hu Yaqian, Wang Shanshan, Zhang Tong, Yuan Jiali, 胡

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In recent years, with the introduction of the concept of metagenomics, human metagenomics has also become a new approach to reveal the development and health of humankind. As the largest microflora in the human body, the gut microbiome has been found to play an important role in the normal physiological functions of the human body and the occurrence and development of diseases. The variety and quantity of gut microbiome are abundant. In the study of gut microbiome, the application of germ-free animals makes the experiment more accurate and convenient. It is an ideal basic animal model. Not only that, models developed on this basis are also of great significance to this type of research. This review summarizes the understanding of germ-free animals, the application of germ-free animals in experimental research and the prospects of germ-free animal models, to review the application and development of germ-free animals in the gut microbiome.
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Volume 32,2024 Issue 10

Volume 32,2024 Issue 10

Volume 32,2024 Issue 10

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