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Volume 34,2024 Issue 10

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Evaluation study on zebrafish insomnia model by illumination method and combined illumination and caffeine method

san yuqing

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【】 Objective: to establish different sleep deprivation models using zebrafish to provide more reproducible and practical modeling reference solutions for basic research on insomnia. Methods: Zebrafish insomnia model was induced by two interventions: continuous light (150 lux) and light plus caffeine, and the zebrafish were randomly divided into control, light, combined and caffeine (100 μmol/l) groups. The difference in locomotor ability of zebrafish in each group was observed by using open field experiment and circadian rhythm behavioral experiment; the expression and secretion of related sleep genes and 5-HT neurotransmitter were detected by using Qpcr technology and Elisa technology. Results: 1. The sleep time of zebrafish in the light group was significantly reduced compared with that of the blank control group (P=0.01), and there was no statistically significant difference between the sleep time of zebrafish in the combined group and the caffeine group (see Table 1); the resting time of zebrafish during the daytime in the light group was significantly reduced compared with that of the blank control group (P<0.01), and the resting time during the daytime of zebrafish in the combined group and the caffeine group was increased (P<0.01) (Table 3). There was no statistically significant difference in the black light resting time of zebrafish in all groups (Table 3); the daytime moving distance of zebrafish in the light group was significantly increased compared with that of blank control group (P < 0.01), and the 10-h activity distance of zebrafish in the light group in the dark day was significantly increased compared with that of blank control group (P < 0.01), and there was no statistically significant difference in the dark day activity distance of zebrafish in the combined and caffeine groups (Table 4); and the sleep rounds of zebrafish in the light group was blank control group was significantly reduced (P < 0.01), and the sleep rounds of zebrafish in the combined and caffeine groups were significantly increased (P < 0.01) compared to the blank control group (Table 2).2. The percentage of swimming distance in the central region of the light group was reduced (P < 0.05) compared to the blank group, while the percentage of swimming distance in the central region of the caffeine group was significantly reduced (P < 0.01) compared to the light group (see Figure 6); the light group The percentage of swimming time in the central region was lower than that in the blank group (P < 0.05), while the percentage of swimming time in the central region was significantly lower in the caffeine group than that in the light group (P < 0.01) (Fig. 7).3. The expression of HTR1aa mRNA level in zebrafish in the light group was higher than that in the blank group at 6:00 a.m. and 12:00 p.m. (P < 0.05) (Figs. 10 and 11), and the expression of There was no statistically significant difference in the expression of HTR1ab mRNA levels between the light group and the combined group (P > 0.05) (Figs. 12 and 13); compared with the blank group, the amount of 5-HT secretion in the light group was reduced in both groups at 6:00 a.m. (P < 0.01), and at 12:00 p.m. the amount of 5-HT was reduced in both the light and combined groups (P < 0.01), and the amount in both groups remained lower than that in the blank group at 6:00 p.m. (P<0.01).
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The effects of combined exposure to PM2.5 and high-salt diet on liver inflammatory cytokines and lymphangiogenesis in mice

ding shibin, JIANG Jinjin, LI Yang

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Objective The effects of combined exposure to PM2.5 and high-salt diet on hepatic inflammation and lymphangiogenesis in mice. Methods Thirty-two C57BL/6J mice were randomly assigned to: control group, PM2.5 group, high-salt group and PM2.5+high-salt group. Mice in the high-salt group and PM2.5+high-salt group were fed with 8% high-salt diet for 8 weeks, mice in the other groups were fed with control diet containing 0.4% salt. Mice in the PM2.5 group and PM2.5+high-salt group were treated with PM2.5 by tracheal instillation (twice per week); and mice in the mice in the other groups were instilled with equal volume of saline at the same time (twice per week). All mice were sacrificed after the last PM2.5 exposure. TNF-α and IL-6 in liver tissues of mice were determined. Moreover, the LYVE1 expression of liver tissues were?visualized using immunofluorescence staining. The protein expression levels of lymphangiogenesis markers PROX1 and LYVE1, lymphangiogenesis regulatory proteins VEGFR-3 and VEGF-C in liver tissue were measured using Western blot. Results Compared to the control group, the levels of TNF-α and IL-6, as well as the protein expressions of PROX1，LYVE1, VEGFR-3 and VEGF-C in liver tissues of mice in the high-salt group (HSD) were obviously increased (P<0.05). Compared to the HSD, the levels of TNF-α and IL-6 and the protein expressions of PROX1，LYVE1, VEGFR-3 and VEGF-C in liver tissues of mice in the PM2.5+HSD were obviously increased (P<0.05). Moreover, there were significant interaction effects between PM2.5 and high-salt diet on these above indicators. Conclusions Combined exposure of PM2.5 and high-salt diet obviously aggravated hepatic inflammation, and may increase hepatic lymphangiogenesis through upregulating VEGFR-3 and VEGF-C in the liver of mice.
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The application value of dual energy CT imaging in the evaluation of bone repair

sudanyang, mayuanbo, liujinlong, zhanghaoran, yangshenyu, miaoqiuju, baizhen, yangxiaopeng

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Bone defect repair is an urgent problem to be solved in the field of orthopedics, and numerous researchers are working to develop more effective treatment plans. However, accurate evaluation of bone repair after surgery is an crucial step. With the development of computed tomography (CT) imaging, dual energy CT imaging has shown significant advantages in analyzing bone composition and reducing metal artifacts. This article reviews the application value of dual energy CT imaging in the evaluation of bone repair in animals.
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MiRNA investigation of the regulatory effects of Allium guajava and Allium cepa sempervirens soup on the Epsin1-FGFR1 pathway in atherosclerotic mice

wang zhao bo, pan yi, lin qian

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: This study aimed to evaluate the therapeutic effects of Gualou Xiebai Banxia Decoction (GXBD) on atherosclerosis (AS) in a mouse model and to explore its underlying mechanisms. Male APOE-/- mice were used to induce the AS model through a 16-week high-fat diet. The mice were randomly divided into a model group, a control group, different dosage groups of GXBD, and a lipid-lowering drug group. Histological analysis using HE and Oil Red O staining was performed to observe changes in arterial walls and plaque formation. The results demonstrated that GXBD significantly reduced plaque formation. Further miRNA sequencing analysis revealed that GXBD regulated multiple AS-related miRNA genes, influencing the expression of key proteins such as Epsin1 and FGFR1, thereby exerting potential therapeutic effects on AS lesions. Western blot and immunofluorescence experiments further validated these regulatory effects. The findings suggest that GXBD exhibits significant efficacy in the treatment of atherosclerosis, potentially through miRNA regulation and protein expression modulation.
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Effects of RSPO2 on oxidative stress and apoptosis of ovarian granulosa cells

LI Shuo, LI Nian, ZHOU Xiaofeng, YUAN Xiaolong

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Objective Utilizing human ovarian granulosa tumor cells as the cellular model, we tried to investigate the effect of roofplate-specific spondin 2 (RSPO2) on the oxidative stress and apoptosis level of ovarian granulosa cells. Methods We performed transient transfection of RSPO2 overexpression plasmid and small interfering RNA (siRNA) in COV434 cells. Real-time Quantitative PCR (RT-PCR) and Western blot were used to detect efficiency of overexpression and interference. The reactive oxygen species assay kit was used to determine the amount of intracellular reactive oxygen species. The apoptosis level was detected by flow cytometry. RT-PCR and Western blot were used to determine the expression levels of oxidative stress and apoptosis-related genes. Co-immunoprecipitation was used to detect interacting proteins. Results The overexpression and interference of RSPO2 were successfully realized. Overexpression of RSPO2 significantly inhibited the level of reactive oxygen species in COV434 cells (P < 0.05) and extremely significant inhibited the early apoptosis of granulosa cells (P < 0.01), interference of RSPO2 obtained the opposite results. Overexpression of RSPO2 significantly upregulated the mRNA and protein levels of SOD1, SOD2, CAT (P < 0.05), and significant downregulated the levels of caspase 3 and caspase 8 (P < 0.05). On the contrary, interference of RSPO2 significantly downregulated the levels of SOD1 and CAT (P < 0.05), and significantly upregulated the level of caspase 3 and caspase 8 (P < 0.05). We found the protein interactions between RSPO2, SOD1, CAT and caspase 3. Conclusion Interfering with RSPO2 promotes the accumulation of reactive oxygen species within cells, downregulates the expression of SOD1 and CAT, thereby enhancing oxidative stress. Additionally, interfering with RSPO2 upregulates the expression of caspase 3 and caspase 8, promoting cell apoptosis. These findings suggest that RSPO2 plays a crucial role in ovarian granulosa cells.
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Current research status of ferroptosis in endometriosis

pengjiahua, song xin, liangruining, sun yu, jian hui, luoxiaoquan

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Endometriosis (EMs)? is a common challenge in global women"s health. In recent years, ferroptosis, as an emerging form of regulated cell death, has attracted attention in EMs research. In terms of disease mechanisms, ferroptosis drives ovarian EMs fibrosis through iron overload induction, and promotes disease progression by regulating multiple signaling pathways such as p38MAPK/STAT6, enhancing angiogenesis, autophagy, etc. In terms of infertility, ferroptosis affects embryo development and ovarian function, significantly reducing fertility. In terms of diagnosis, the high expression of ferroptosis-related genes provides potential biomarkers for early diagnosis of EMs, effectively distinguishing patients from healthy individuals, and has important clinical application value. In terms of treatment strategies, non-natural compounds like Erastin, as well as natural compounds like resveratrol, ursolic acid, baicalein, etc., show potential therapeutic effects, relieving the pathological process and related symptoms of EMs. Therefore, this article
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Research progress of the ketogenic diet in the field of Parkinson"s disease

LI Boyang, LAN Rui, YANG Huijie, LI Hongyu, LI Chiyang, SHEN Xiaoming

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A ketogenic diet refers to an eating pattern designed to promote individuals to achieve a low-calorie content, minimum carbohydrate intake, high-fat consumption, and standard protein levels. The ketogenic diet is used in clinical practice to treat applications including heart dis-ease, diabetes, obesity, autism, glioblastoma, and other cancers. However, the ketogenic diet has not been recommended for any neurological disorder except epilepsy. With the intensive research on the ketogenic diet in recent years, more and more evidence has proved that the ketogenic diet has a neuroprotective effect on Parkinson's disease, and it is a new dietary ther-apy for the treatment of Parkinson's disease. In this review, we discuss in detail the neuropro-tective effects of the ketogenic diet in Parkinson's disease by different mechanisms, aiming to provide some references for clinical and experimental studies in this field.
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Study on repairing effect of moxibustion at different acupoints

zhouhaiyi, hesiyi, hanruifang, guanyongge, donglijuan, Song Yang

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Objective: To compare the repair effects of moxibustion "Guanyuan" and "Shenshu" on thin endometrium rats. Methods: 32 SD rats were randomly divided into normal group, model group, Guan Yuan group and Shenshu group with 8 rats in each group. Thin endometrium model was established using 95% anhydrous alcohol. After two estrus cycles, the Guanyuan group and Shenshu group moxibusted "Guanyuan" and "Shenshu" points, once a day, and rested for 1 day every 6 days for a total of 21 days. The other two groups were fixed in the same way. The temperature changes in the acupoint area at different time were measured by infrared thermometer, the pathological morphology of uterus was observed by HE staining, and serum levels of IL-1β, IL-10, E2 and PROG were detected by ELISA. The expressions of Ki67, PCNA, LIF and ITG-β3 in uterus were detected by immunohistochemistry. The expression levels of ANG-1、CD34和VEGFA in uterus were detected by Western blot. Results: Immediately after moxibustion and 5min after moxibustion, the body surface temperature at Shenshu point was higher than that at Guanyuan point (P<0.001, P<0.01). Compared with normal group, endometrial thickness, number of glands and blood vessels in model group were decreased (P<0.0001, P< 0.01, P<0.001), The endometrial thickness, number of glands and blood vessels increased in Guanyuan group (P<0.0001, P<0.01, P<0.01) and Shenshu group (P < 0.0001, P<0.01, P<0.05),and the endometrial thickness, number of glands and blood vessels in Guanyuan group were better than those in Shenshu group, but the difference was not statistically significant. Compared with normal group, the serum IL-1β level in model group was increased (P<0.01), and the PROG level was decreased (P<0.001). Compared with model group, the serum IL-1β in Guanyuan group were decreased (P<0.01), E2 level and PROG level increased (P<0.01, P<0.05),and the serum IL-1β were decreased (P<0.05), IL-10,E2 and PROG levels (P<0.05, P<0.05, P<0.01) were increased in Shenshu group. Compared with normal group, the expressions of Ki67, PCNA, LIF and ITG-β3 of model group were decreased (P< 0.0001). Compared with model group, the expressions of Ki67, PCNA, LIF and ITG-β3 in Guanyuan group (P<0.0001) and Shenshu group (P<0.001, P<0.001, P<0.0001, P<0.001) were increased, and PCNA expression in Guanyuan group was higher than that in Shenshu group (P<0.05). The expressions of LIF and ITG-β3 in Shenshu group were stronger than those in Guanyuan group (P<0.01, P<0.05). Compared with normal group, the expression of ANG-1 and CD34 protein in uterine tissue of model group was decreased (P<0.0001). Compared with model group, the expressions of ANG-1, CD34 and VEGFA in Guanyuan group were increased (P<0.0001), and the expressions of CD34 and VEGFA in Shenyu group were increased (P<0.0001). The expression of ANG-1 in Guanyuan group was higher than that in Shenyu group (P<0.0001), while the expression of CD34 in Shenyu group was higher than that in Guanyuan group (P<0.05). Conclusion: Moxibustion "Guanyuan" point and "Shenshu" point could repair thin endometrium, Guanyuan point is superior to Shenshu point in reducing the level of inflammatory factors, increasing the level of E2, promoting the expression of proliferation factors, while Shenshu point is superior to Guanyuan point in increasing the level of anti-inflammatory factors and PROG, and promoting the expression of endometrium receptor related factors. The two points may play a role in promoting angiogenesis through different ways.
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Preliminary Study on the Application of Three-Dimensional Time-of-Flight Magnetic Resonance Angiography (3D-TOF-MRA) to Observe the Cerebral Arteries in Mongolian Gerbils

JIAO Kun, ZHANG Jing, MENG Xia, WANG Zhanjing, LEI Jianfeng, CHEN Baian, LU Jing

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Objective Establishment of a three dimensional time-of-flight magnetic resonance angiography (3D-TOF-MRA) technique for observing cerebral arteries in live Mongolian gerbils. Method 1. Employ a 7.0T MRI scanner to image the cerebral arteries of Mongolian gerbils, with data subsequently processed using the RadiAnt DICOM Viewer software；2. Validate and compare imaging results via the latex perfusion method；Attempt to perform 3D reconstruction of the posterior communicating branch and surrounding cerebral blood vessels of the Mongolian gerbil and rats through analysis software; 4. Attempt to use this method to screen individuals with abnormal vascular development in Mongolian gerbils to verify the effectiveness of this method. Result 1.The 3D-TOF-MRA technology can effectively observe the cerebral arteries of living Mongolian gerbils; 2.The 3D-TOF-MRA technology has high accuracy in observing the main cerebral arteries of Mongolian gerbils, but its observation effect on finer vascular branches is not as good as latex perfusion method; 3.The data obtained from 3D-TOF-MRA technology can be used for three-dimensional reconstruction of blood vessels; 4. The application of this technology has a good effect on screening for abnormal development of arterial structure in Mongolian gerbils. Conclusion The 3D-TOF-MRA technology can be applied to the structural observation and related research of the cerebral arteries in live Mongolian gerbils.
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Macrophage polarization in the research development of renal ischemia-reperfusion injury

SUN Haojie, LIU Xianqin, WANG Suogang ?

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Macrophages are important immune cells in innate immunity, with significant heterogeneity and polarization. It can polarize into various phenotypes (mainly M1 and M2) under the stimulation of various factors in the microenvironment, thus exerting different roles and functions. It plays an important role in the pathophysiological mechanism of renal ischemia-reperfusion injury (RIRI). Excessive immune response will inevitably lead to tissue damage. M1 macrophages are pro-inflammatory cells, which are involved in the clearance of pathogens. M2-type macrophages have anti-inflammatory effects and participate in the repair and remodeling of renal tissue after RIRI. The balance between macrophage phenotypes is important for the outcome and treatment of RIRI. Reason from the macrophage polarization Angle, the macrophages in RIRI pathophysiologic mechanism and the latest treatments, designed to further study the function of macrophage polarization in RIRI for reference, to adjust the macrophage polarization to improve RIRI treatment strategies.
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Research progress of TAM in tumorigenesis, development and drug resistance of gastric carcinoma

Cai Qiuling, Yang Jing, Shen Huiling, xuwenlin

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Tumor-associated macrophages(TAM) are a current focus in the study of the tumor microenvironment. To achieve their functionally distinct roles, macrophages undergo phenotypic polarizations resulting in two major subgroups: M1 macrophages with pro-inflammatory and anti-tumor effects, and M2 macrophages with anti-inflammatory and pro-tumor effects. Among them, M2 polarization, which is the main manifestation of TAM, has been associated with poor prognosis in various cancers and has shown supportive properties for malignancies. Gastric carcinoma patients have a low early diagnosis rate, late disease stage, and poor prognosis, with biological behavior characteristics of easy recurrence and metastasis. Currently, drug resistance and toxic side effects significantly limit the application and effectiveness of gastric carcinoma treatment. Therefore, it is urgent to explore new therapeutic drugs or targets. We summarize in this review the recent progress in the studies of TAM in the occurrence, development and drug resistance of gastric carcinoma, providing new ideas for clinical treatment and prognosis analysis of patients with gastric carcinoma.
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Construction of Lep gene knockout mouse model based on CRISPR/Cas9 system

CAO YUAN, YANG YUANSONG, GU WENDA, ZHAO HAOYANG, ZHAI SHIJIE, SUN XIAOWEI, FAN CHANGFA

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Objective The ob mice (C57BL/6-Lepem1/Nifdc) with the Lep gene knocked out mice (ob/ob) were generated by CRISPR/Cas9 system, establishing an animal model suitable for preclinical drug evaluation in clinical diseases such as diabetes. Methods According to the principle of CRISPR/Cas9 target design, sgRNA targeted mouse Lep gene was designed for transcription in vitro, and micoinjected with Cas9 mRNA into mouse zygotes. Mice tails DNA was extracted and detected by PCR and sequencing, followed by mating positive mice and wild-type mice. Blood biochemistry and liver pathology were assessed in homozygous ob mice. Results 8 positive mice were identified and a stable mouse strain was selected for further breeding. The serum triglycerides (TG), total cholesterol (CHO) and alanine aminotransferase (ALT) levels in homozygous ob mice were significantly higher than wild-type mice. Liver pathology result showed significant inflammatory infiltration and lipid vacuolar transformations. Conclusion The successful establishment of Lep gene knockout mice has enriched the national rodent experimental animal database and provided an animal model for preclinical drug evaluation.
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The role of astrocytes in traumatic brain injury and therapeutic strategies

Huang Yizhen, Cheng hao, Wang Hao-wei, Zhang Qian-yao, Luo Cheng-liang, Zeng Xiao-feng

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Traumatic brain injury (TBI) is an injury caused by the direct or indirect effects of external factors that result in structural or functional loss of brain tissue. Astrocytes, as homeostatic cells of the central nervous system, proliferate and activate rapidly in the early stages of TBI, and participate in a series of pathological processes such as neuroinflammation, blood-brain barrier disruption, glial scarring, and excitotoxicity after injury, and play a crucial role in the secondary neurological injury of TBI. This paper reviews the role of astrocytes in the repair of TBI damage with the aim of providing new strategies for the prevention and treatment of TBI.Traumatic brain injury (TBI) is a kind of craniocerebral injurious disease caused by external force that results in structural destruction of brain tissue and physiological dysfunction, with high morbidity, high disability, high mortality and high treatment burden, for which there has not yet been an effective therapeutic measure. Astrocytes, as homeostatic cells of the central nervous system, are rapidly proliferated and activated in the early stages of TBI to participate in a series of pathological processes such as neuroinflammation, blood-brain barrier disruption, glial scarring, excitotoxicity and other pathological processes after injury, and play a crucial role in the secondary neurological injury of TBI. The article focuses on the role of astrocytes in the pathological damage of TBI and reviews the research progress of astrocyte-targeted drugs, aiming to provide new strategies for the prevention and treatment of TBI.
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Research progress on anti-tumor effects of LRRC8A/VRAC ion channels

yangrunze, huyaohua, qinjing, wangyongfeng, shichanghong

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Volume adjusting anion channel (VRAC) in vertebrate cells and each type of general expression in tumor cells. Leucine-rich repeat containing 8A(LRRC8A) and its four homologous family members (LRRC8B-E), of which LRRC8A is an essential subunit. It has been confirmed that LRRC8A/VRAC is involved in the proliferation, migration, invasion and multi-drug resistance of tumor cells through various signaling pathways. This ion transporter has shown good potential in killing tumor cells and preventing the development of tumor, and can be used as a new target for tumor therapy. Therefore, this paper reviews the latest research on the involvement of LRRC8A/VRAC in tumorigenesis and development, including the molecular structure, function and regulation of LRRC8A/VRAC in tumor and immune cells, with emphasis on the application of LRRC8A/VRAC in tumor diagnosis and immunotherapy, in order to provide reference for exploring LRRC8A/VRAC as a new target of tumor therapy.
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The role of PI3K/AKT signal pathway in peripheral nerve injury and the research progress of traditional Chinese medicine intervention

guo xiaojing, ZHANG bo, Wang Yan, ZHANG Li

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Peripheral nerve injury (PNI), caused by contusions, fractures, and other traumas, leads to abnormal sensory functions, limited motor capabilities, neuropathic pain, and muscle atrophy, severely impacting patients' quality of life. Post-PNI, Wallerian degeneration occurs, involving axonal degeneration and myelin sheath collapse. Notably, the dorsal root ganglia (DRG), as the primary sensory neurons in pain signal transmission, are crucial targets in neuropathic pain (NP) induced by PNI. Therefore, changes in these targets trigger a series of complex signaling pathway alterations. Among them, the phosphatidylinositol 3-kinase/serine-threonine kinase (PI3K/AKT) signaling pathway, a vital regulator of cell survival and death, promotes Schwann cells (SCs) proliferation and migration, thereby enhancing axonal growth and myelination to facilitate nerve regeneration, and supports the survival of DRG neurons to alleviate NP. Current treatment methods include stem cell transplantation and neurotrophic medications, but all have certain limitations. Studies have shown that traditional Chinese medicine (TCM) has gained increasing attention and has been applied to some extent in the treatment of PNI due to its advantages of low cost and fewer side effects. This article discusses the relationship between oxidative stress, apoptosis, autophagy, inflammation, angiogenesis, the cell cycle, and other pathophysiological mechanisms in PNI and the PI3K/AKT signaling pathway, as well as its associated molecular targets. It also delves into the potential mechanisms of action of TCM monomers, compound formulas, and acupuncture based on the PI3K/AKT signaling pathway in the treatment of PNI, aiming to provide systematic and standardized theoretical guidance for the healing of PNI with TCM and a useful reference for the development of related medications.
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Research progress in the mechanism of PPARγ gene in osteoarthritis

Zhang Jian, Wang Dong, Qi Tiantian, Wu Liangbin, Yang Qi, Yu Fei

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Osteoarthritis (OA) is a chronic degenerative disease with a high incidence rate among middle-aged and elderly people. In the early stage, OA can be treated by conservative methods. In the late stage, most patients choose joint replacement and other surgical methods to treat OA, causing pain to patients and their families. Peroxisome proliferator activated receptor γ (PPAR γ) is a gene discovered in recent years that plays an important role in the degeneration or inflammation of OA cartilage, synovium, adipose, and other tissues. Signaling pathways such as AMPK, Wnt, and SIRT1 may be involved in this process. This review focuses on the role and mechanism of PPARγ gene’s function on OA via joint related tissues and signaling pathways.
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Biogenesis of exosomes and their role in neurodegenerative diseases

GuoSixu, MeiXinxin, LiJin

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The incidence of neurodegenerative diseases (NDs) is increasing with the years, and current drugs targeting NDs are basically symptom-based, not therapeutic, so they can’t prevent disease progress. Exosomes are a subset of extracellular vesicles that can be produced by all cells in the human body and exert biological effects in a variety of ways after they are released into the extracellular environment. Many studies have shown that although exosomes have two sides in the central nervous system, they can still be used as a tool for the prevention, remission, treatment and diagnosis of NDs due to their unique biological characteristics. This article reviews the biogenesis of exosomes and their roles in neurodegenerative diseases, so as to provide new ideas for clinical treatment of NDs.
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Research progress on hypoxic cell models

LI Jing, XU Dongyang, LI Changqing, SU Mengyao, WANG Zhijuan, ZHAO Mingjun, ZHAO Jialong, YANG Junyi, YANG Qiaodie, Kang Longli

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Hypoxia is associated with the occurrence and development of many diseases in clinical settings. Cell hypoxia not only serves as a vital marker for disease advancement but also plays a pivotal role in exacerbating the disease process. Therefore, improving tissue hypoxia may provide new strategies for the treatment of related diseases.. To investigate these diseases at the cellular and molecular levels, it is paramount to establish a cellular hypoxia model. At present, the extensively employed hypoxic cell models can be primarily categorized into three types: chemical hypoxia model, physical hypoxia model, and glucose deprivation hypoxia model.This article will overview the various types of hypoxic cell models and scrutinize their application and limitations in disease research.
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Vascular smooth muscle cell phenotype switch and Cell pyroptosis in the role and research progress of Abdominal aortic aneurysm

Liu Xin, Tang hongyue, Guo chang, Ma dong, Zhang mingming

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Abdominal aortic aneurysm (AAA) is a hidden and fatal disease, but its underlying developmental mechanism remains unclear. It has been found that phenotypic switching of vascular smooth muscle cells and pyroptosis are two biological processes closely related to the appearance and progress of AAA, which may become an important theory to reveal the mechanism of AAA and provide a new direction for the diagnosis and treatment of AAA. In this review, we will discuss the phenotypic switch of VSMCs and the regulatory relationship between cell pyroptosis and AAA.
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Research Progress of microRNA in the Field of Deep Vein Thrombosis

LILei, YUAN Qidong, PENG Xitao, ZHU Jin, PENG Juncai, HE Changhai, FU Liqing

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Micro-ribonucleic acid (miRNA) is a class of endogenous RNA molecules with a length typically ranging from 19 to 25 nucleotides.They regulate gene expression levels by identifying homologous sequences and intervening in transcription, translation,or epigenetic processes.miRNAs exhibit potential application value in the pathogenesis, progression,and treatment of deep vein thrombosis (DVT).DVT refers to the abnormal coagulation of blood within the lumen of the deep veins,blocking the venous lumen and leading to venous return obstruction, which is more common in the lower limbs.The detachment of thrombus into the lungs can lead to death.This article comprehensively reviews the diverse mechanisms of action of miRNAs in DVT in recent years.Given that the regulation of miRNA expression through targeted therapeutic approaches may promote the recovery of DVT,this article also discusses in detail the potential applications of miRNAs in the clinical diagnosis and treatment of DVT.This article aims to provide valuable references and insights for clinical and basic research in the field of DVT.
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Recent progress of research into the role of lncRNA in the molecular mechanism of pulmonary hypertension

WU Daoxiong, LI Yanjin, HU Ying, WANG Yuming, HU Wei, MA Run

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Pulmonary hypertension (PH) is a fatal disease characterized by pulmonary vascular remodeling that ultimately leads to right heart failure and death. Current PH treatment is suboptimal, with no substantial improvement in the overall survival of patients with advanced PH. Despite the current partial progress has been made in the pathogenesis of PH, more exploration at the molecular level is needed to develop more effective treatments for PH. Recent researches have shown that long non-coding RNA (lncRNA) plays an important regulatory function in the pathophysiological process of PH, and has become a potential disease biomarker and therapeutic target. In this paper, we review the molecular mechanisms of lncRNA in PH in recent years.
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Research progress of Wnt/β-catenin signaling pathway in programmed cell death after ischemic stroke

Panrui, Zhangming, Zhengxinyue, Wangchunxiao, Renqiongdi

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【】 Ischemic stroke (IS) is a neurological disease that damages brain tissue from insufficient blood supply to the brain due to blockage or stenosis of the brain vessels. In recent years, there has been increasing evidence that the Wnt/β-catenin signaling pathway plays an important role in the pathophysiological response to the occurrence and development of IS. Programmed cell death includes many forms such as apoptosis, necrotic apoptosis, pyrodeath, autophagy, panapoptosis and iron death. In this review, we will elucidate the characteristics of these different cell death modes and their cross-talk relationships with each other, and systematically outline the role of Wnt/β-catenin signaling pathways in the intervention of different cell death modes in IS, aiming to provide reference for future clinical and basic research.
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Comparison of stabilizing effect of different dose-bleomycin-induced pulmonary fibrosis in rats

songqinghua, tanghuimeng, sunxin, liuyang, xieyunyun, tianyange

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Objective: To compare the success rate and stability of rat pulmonary fibrosis (PF) models induced by intratracheal instillation of different doses of bleomycin (BLM). Methods: 150 SD rats were randomly divided into control group, low-dose BLM 3 mg/kg group (BL-L group), and high-dose BLM 5 mg/kg group (BL-H group). The general status, mortality, and weight changes of the rats were observed, the inspiratory capacity (IC), vital capacity (VC), chord compliance (Cchord) and dynamic compliance (Cdyn) of the lungs were detected on the 28th, 42nd, 56th and 84th days. Lung coefficients were recorded, pathological changes in lung tissue were observed by HE and MASSON staining, and collagen type III (COL III) expression in lung tissue was detected by immunohistochemistry. Results: (1) Mortality rate: The mortality rates of BL-L group and BL-H group were 20% and 28%. (2) Weight: The weight of the BL-L group was lower than that of the control group on days 0-56, and the weight loss was most significant on day 14 (P < 0.01), with weight recovery after day 56. From day 0 to day 56, the weight of the BL-H group was significantly lower than that of the control group and BL-L group (P < 0.01), and the weight gain remained relatively slow after day 56. (3) Lung function: compared with the control group, on day 28, the lung function of IC, VC, Cchord, and Cdyn were significantly lower in the BL-L group (P < 0.01, P < 0.05), and IC and Cdyn were significantly lower in the BL-H group (P < 0.01). On day 42, IC, VC, and Cchord decreased significantly in the BL-L group (P < 0.01, P < 0.05), while IC, VC, Cchord, and Cdyn significantly decreased in the BL-H group (P < 0.01, P < 0.05), and IC, VC, and Cchord were significantly lower compared with those in the BL-L group (P < 0.01). On day 56, Cchord was significantly lower in the BL-H group than in the control and BL-L groups (P < 0.01), and on day 84, there was no significant difference in lung function between the groups. (4) Lung coefficient: on day 28, the lung coefficients of the BL-L and BL-H groups were significantly higher than those of the control group (P < 0.01), and from day 42 to 56, the lung coefficients of the BL-H group were significantly higher than those of the BL-L group and the control group (P < 0.01), and on day 84, the lung coefficients of the groups did not differ significantly from those of the control group. (5) Lung histopathology and immunohistochemistry: inflammatory infiltration, fibrotic streaks and COL III expression were observed in the BL-L group from day 28 to 56, with the severity of the lesions decreasing over time, and the fibrotic lesions had almost disappeared on day 84, while in the BL-H group, significant fibrotic lesions could be observed from day 28 to 84, and the degree of COL III expression was significantly higher than that in the control group (P < 0.01). Conclusions: Both 3 mg/kg and 5 mg/kg BLM can successfully induce PF rat models. The PF model prepared with 3 mg/kg BLM developed fibrosis on day 28, which lasted until day 42 and gradually recovered. The PF model prepared with 5 mg/kg BLM showed fibrosis formation on day 28, and the degree of fibrosis was more severe than in the lower dose, with fibrotic lesions persisting stably up to day 56, and moderate-to-severe fibrosis still present in half of the cases until day 84.
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Research progress on the evaluation and intervention of social interaction behavior in animal models of autism

KONG Minghui, LU Liming, XIANG Leiying, CHEN Xiaoyi, ZHU Zhiru

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Autism Spectrum Disorder (ASD) is a highly heterogeneous neurodevelopmental disorder with a complex underlying genetic structure. However, preclinical trials currently under investigation for ASD mainly rely on rodent models to test the effects of non–pharmacological and pharmacological interventions on the core and related symptoms of ASD. This article elaborates on the brain regions that affect social interaction behaviors from the perspective of cognitive neural mechanisms; reviews behavioral testing experiments such as the three–chamber social interaction test, the visible burrow system, and the ecological HAB system; summarizes effective non–pharmacological interventions and pharmacological interventions such as baclofen, oxytocin, and metformin in the core and related symptom areas of ASD. This review aims to provide reference directions for promoting the development of preclinical trials using rodent models in the future.
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Study on the Pathology of Gouty Arthritis Based on Purinergic Receptor P2X7R

Wang Yinfeng, Liu Zeyu, Yin Xiaoyang, Lu Chengjin, Lin Zhijian, Zhang Bing

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Abstract: Objective: To observe the effects of Ice-Water Swimming on pathological changes in gouty rat model and investigate the regulatory mechanism of P2X7R therein. Methods: Male SD rats were divided into Normal group and Experimental groups, which included Gouty Control group, Ice-Water Swimming group and BBG group. Experimental groups were modeled to hyperuricemia and gouty arthritis by inhibiting uric acid metabolism combined with Coderre method. Ice-Water Swimming group were treated with 5 min of endurance swimming in an ice-water mixture at a depth of about 0.5 m for 0 h and 12 h after modeling by Coderre method, while the BBG group were injected intraperitoneally with BBG solution for 1 time immediately after modeling. Formula to calculate ankle swelling index. The serum uric acid level of rats was detected by uricase assay. The expression level of serum inflammatory factors IL-1β, IL-6 and TNF-α was detected by ELISA. HE staining was used to observe the pathological state of the subject ankle joints. Western Blot and Immunohistochemistry for P2X7R and NLRP3 Protein Expression in Synovial Tissue respectively. Results: Compared with Normal group, the serum uric acid level and ankle joint swelling index of Experimental groups were significantly higher (P < 0.05 or P < 0.01), and the synovial tissues all had different degrees of inflammatory infiltration. Compared with Gouty control group, the ankle swelling index of Ice-Water Swimming group was significantly higher at 12h (P < 0.05). The serum IL-1β, IL-6, TNF-α levels (P < 0.01) and the protein expression of P2X7R, NLRP3 in the synovial tissues were all significantly elevated (P < 0.05). Pathology results showed that the cartilage surface was broken. In addition, the synovial tissue showed severe hyperplasia and erosion, accompanied by a large number of inflammatory cell aggregates. There were no significant changes in P2X7R, NLRP3 protein expression and pathological damage in synovial tissues of BBG group compared with Gouty control group (P > 0.05), whereas the expression of serum IL-1β, IL-6, and TNF-α were all significantly suppressed (P < 0.01). Conclusion: Cold stimulation and strenuous exercise simulated by Ice-Water Swimming can exacerbate pathological damage in gouty arthritis by a mechanism that may be related to high localized P2X7R expression in the joints.
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Research progress on animal models of insomnia based on combination of disease and syndrome

ZHAO Zhongmin, LIANG Shuzhi, GAO Xiangye, HOU Suyu, LUO Dandan, WANG SHIjun, ZHANG Yan

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Insomnia, a prevalent clinical condition, not only diminishes patients" quality of life but also has the potential to give rise to further health complications. Traditional Chinese medicine has amassed and passed down knowledge about treating insomnia for thousands of years, resulting[1] in distinct advantages. It utilises syndrome differentiation and therapy to accomplish therapeutic effects by regulating the internal balance of the human body. To get a deeper understanding of how Traditional Chinese Medicine (TCM) treats insomnia, it is crucial to create animal models that exhibit symptoms closely resembling those found in humans with insomnia. This article provides a concise overview of the current disease-syndrome combination models, which may be classified into four categories: liver-depression, liver-depression for fire or liver-depression for spleen deficiency, heart-spleen deficiency, heart-kidney incompatibility, Yin and blood deficiency, This discussion will focus on particular modelling methodologies, assessment indicators, and model performance. The aim is to enhance the understanding of the pathophysiology of this disease in the context of traditional Chinese medicine. Additionally, this discussion aims to provide technical assistance for clinical efficacy evaluation and the creation of novel drugs.
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Current situation and countermeasures of laboratory animal license management in Hubei Province

liuconglin, qiaochuhua, liyanmei, chenxiaoli, zhangjinming, liudan, chenrui

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This paper introduces the daily management service of experimental animal license in Hubei Province and the supervision matters during and after the event. It expounds the current situation of experimental animal license management, achievements, problems and countermeasures in Hubei Province.The focus was on analyzing the legislative situation of experimental animals in Hubei Province, the issuance and distribution of permits, the scale of facilities, and the composition of employees. In recent years, the number of experimental animal licenses issued in Hubei Province has been increasing year by year, and the number of animal production and use by licensed units has continued to rise. While the related industries are flourishing, there are also some regulatory deficiencies. this paper from the perspective of biosafety, combined with the problems encountered in the license management service work, put forward the relevant safety supervision countermeasures and suggestions, so as to better promote the development of experimental animal industry in Hubei Province.
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Application research progress of single-cell sequencing technology in Multiple Sclerosis

MA Zhihong, ZHANG Lihong, CAO Yuan, CHENG Ming

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Multiple Sclerosis (MS) is a chronic autoimmune disease of the central nervous system (CNS) characterized by neuroinflammation and neurodegeneration. The pathogenesis of these diseases is complex, and a variety of central nervous system and peripheral cells are involved. In recent years, single-cell sequencing has gradually been applied in neurological disorder, which is of great significance for understanding intercellular heterogeneity, disease development mechanisms, and treatment strategies. This review will summarize the single-cell sequencing and its applications in MS.
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Mechanism study of mechanical stress mediated KOA synovial fibrosis through Piezo1/ERK1/2 axis

YuLikai, TianDi, SuZishan, JieLishi, GuoShaobo, WangPeimin, ZhangNongshan

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Objective To investigate the effect of Piezo1 activated by mechanical stress on KOA synovial fibrosis via ERK1/2 signaling pathway. Methods (1) Animal experiments: 25 SD rats were divided into blank group, exercise group, exercise +GsMTx4 group, exercise +PD98059 group, exercise +GsMTx4+PD98059 group, 5 in each group. After the modeling was completed, the serum and synovial tissue of rats were extracted and the collagen deposition was evaluated by Sirius scarlet staining and Masson staining. Western Blot and RT-qPCR were used to detect the expressions of Piezo1, ERK1/2, p-ERK1/2, α-SMA, TGF-β, Collagen I and TIMP-1, and the contents of IL-1β, IL-6 and TNF-α were detected by ELISA. (2) Cell experiment: the synovial cells were divided into five groups: blank group, pull group, pull +GsMTx4 group, pull +PD98059 group, pull +GsMTx4+PD98059 group. WB, RT-qPCR and other techniques were used to detect the above indexes in the molded cells. Results (1) Animal experiments: Mechanical stress increased Collagen deposition in synovial tissue of rats, and increased the protein and mRNA expressions of Piezo1, P-ERK /Erk, α-SMA, TGF-β, Collagen I and TIMP-1 of pathway-related and fibrosis-specific indicators (P< 0.05). The expression of Piezo1 was significantly down-regulated with both inhibitors (P< 0.05), but the ERK inhibitor (PD98059) had no significant effect on PIEZO1 expression. The content of serum inflammatory factors in exercise group was significantly higher than that in blank group (P < 0.05), and the improvement was obvious after the use of inhibitors (P< 0.05). (2) Cell experiment: WB and RT-qPCR results showed the same trend as animal experiments. Conclusion The Piezo1 ion channel can sense mechanical stress and activate the ERK1/2 pathway to mediate knee synovial fibrosis.
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Design and Application of Energy Monitoring System for Laboratory Animal Facilities

XU Xiao, XIONG Wenjing, MA Xixiang, MU Dandan, ZHOU Shunchang

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Objective The energy consumption of experimental animal facilities accounts for a significant proportion of the energy consumption in scientific research institutions, and its energy consumption characteristics are significantly different from those of ordinary buildings, thus requiring specialized monitoring and management. Method A set of energy consumption monitoring systems was designed for experimental animal facilities, and was deployed in the SPF-level experimental animal facility of Huazhong University of Science and Technology. Results After the system went online, it achieved real-time collection and recording of the facility"s electricity consumption data, and proposed energy-saving measures for three application scenarios. Conclusion The energy consumption monitoring system for experimental animal facilities has the characteristics of being reliable, efficient, and user-friendly, and has guiding significance and promotion value for the energy management work of experimental animal facilities.
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To study the effect of Houpu Sanwu decoction on exosomes of colorectal cancer model mice based on Rab27a

CHEN shu, RONG kang, ZHANG nan

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Objective: TO investigate THE effect and mechanism of Houpu Sanwu Decoction on subcutaneous tumor in C57BL/6J mice. Methods: Mouse colorectal cancer MC-38 cells were subcutaneously injected into C57BL/6J mice to establish a subcutaneous tumor model of colorectal cancer. The mice with successful model were randomly divided into model control group, Houpu Sanwu decoction low-dose group, Houpu Sanwu decoction medium-dose group, Houpu Sanwu decoction high-dose group, 5-fluorouracil group (5-FU group), and combination group (5-fluorouracil + Houpu Sanwu decoction medium-dose group), with 6 mice in each group. The mice were treated with normal saline and Houpu Sanwu decoction by gavage or normal saline and 5-FU by intraperitoneal injection once a day for 24 consecutive days. During the intervention period, the mental state, diet and other general conditions of mice were regularly detected, and the changes in tumor volume of mice were regularly detected. After sampling, the subcutaneous tumors of each group were weighed and the pathological changes of subcutaneous tumors were observed by HE staining Transmission electron microscopy, Western blot and NTA were used to identify the exosomes derived from colorectal cancer tissues. RT-qPCR and WB were used to detect the mRNA and protein expression levels of Rab27a and exosome markers CD63, TSG101, and ALIX in tumor tissues.Results: Houpu Sanwu decoction could inhibit the growth of tumor in mice. The tumor growth was slow in all dose groups of Houpu Sanwu decoction and its combination group, and the tumor growth was inhibited to varying degrees. The anticancer effect of middle dose of traditional Chinese medicine was the best, and the combination group showed synergistic effect with 5-FU. Houpu Sanwu decoction could induce tumor necrosis, showing the same anticancer effect as 5-FU group, and the higher the dose, the more serious the necrosis was. Moreover, Houpu Sanwu decoction combined with 5-FU showed synergistic effect. Houpusanwu decoction can inhibit the protein and mRNA expression of Rab27a and exosome specific markers CD63, ALIX and TSG101 in mice.Conclusion: Houpu Sanwu Decoction inhibits the secretion of exosomes regulated by Rab27a possibly by inhibiting the secretion of exosomes.
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Analysis of an animal model of hepatic encephalopathy based on the characteristics of clinical diseases in Chinese and Western medicine

liyaqing, wangcan, miaomingsan

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Objective To classify and analyze the existing animal models of hepatic encephalopathy based on the characteristics of Chinese and Western medicine clinical conditions, with a view to providing ideas for improving and perfecting the existing animal models of hepatic encephalopathy, so as to prepare animal models of hepatic encephalopathy that better meet the characteristics of the clinical conditions of Chinese and Western medicine combined with the conditions of hepatic encephalopathy. Methods In China Knowledge Network and Wanfang database, the Keywords "hepatic encephalopathy", "hepatic coma", "animal", "animal model" were used as Keywords, and in Pubmed database, "hepatic encephalopathy", "hepatic coma", " animal model", "animal" as the Keywords to search, collate effective literature with specific animal model preparation methods, collate and classify according to different modeling factors and methods, summarize the modeling methods and characteristics, and assess the clinical match of the animal model based on the characteristics of clinical diseases of hepatic encephalopathy in Chinese and western medicine. The clinical match of the animal model was assigned and analyzed according to the clinical symptoms of hepatic encephalopathy in Chinese and western medicine. Results There were 128 valid literature articles with 11 animal models, including 15 different modeling methods, among which, the CCl4 oil solution model had a high degree of agreement, and the highest degree of agreement in Western medicine was the CCl4 oil solution intraperitoneal injection+gavage+ammonium chloride intraperitoneal injection model (92%), and the highest degree of agreement in traditional Chinese medicine was the CCl4 oil solution intraperitoneal injection+ethanol aqueous solution self-drinking model (65%), but there was a lack of Chinese and Western medicine clinical disease characteristics.Conclusion The existing animal models of hepatic encephalopathy are all in accordance with the characteristics of the disease, but generally lack the manifestation of Chinese medicine, and lack of animal models combining the characteristics of Chinese and Western medicine. Combining the clinical characteristics of Chinese and Western medicine in the preparation of animal models is expected to provide a model basis for clinical diagnosis and treatment of hepatic encephalopathy, and point out the direction for the combination of Chinese and Western medicine in the treatment of hepatic encephalopathy.
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Establishment and optimization of a method for transferring golden hamster embryos using true pregnancy receptors

LAI Ya-na, ZENG Wen-tao

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Objective By optimizing the technical parameters of golden hamster embryo transfer, explore the optimal number and developmental period of transplanted embryos for golden hamster true pregnancy recipients. Methods By establishing a population of true pregnant albino golden hamsters as embryo transfer recipients, the effects of different embryonic development stages, recipient embryo reduction, and secondary transfer recipients on litter yield, donor embryo yield, and offspring survival rate were compared. Results Compared to wild-type golden hamsters as transplant recipients, true pregnancy albino receptors can quickly determine the origin of offspring and are suitable for various reproductive experimental programs based on embryo transfer. Fertilized eggs or two cell embryos have no significant impact on the transfer effect (P>0.05); The rate of donor embryos in the recipient embryo reduction group was significantly increased (22%, P<0.05), and the lower total number of births resulted in a significant decrease in cage survival rate (56%, P<0.01). The second transfer recipient"s non pregnancy rate was significantly increased (42%, P<0.01); The highest embryo yield rate (27%, P<0.01) and normal survival rate (89%) were observed in the transfer of 6-10 embryos. Conclusion The transfer of 6-10 donor embryos can help improve the yield and survival rate of donor embryos, We have successfully established an embryo transfer method using albino pregnant golden hamsters as recipients, providing technical support for the application and development of gene modification models in golden hamsters.
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Progress in the study of the regulation of rheumatoid arthritis-related pathways by external treatment of traditional Chinese medicine

ZHAO Ziying, WANG Guangyi

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Rheumatoid arthritis (RA) is an autoimmune disease with symmetric small joints pain as an early clinical manifestation, which gradually worsens and often involves multiple joints, ultimately resulting in arthritic deformities and loss of labor capacity. Molecular signaling pathways have always been the focus of research on the prevention and treatment of RA. Studies have found that the progression of RA disease is closely related to a variety of signaling pathways which include the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway, mitogen-activated protein kinases (MAPKs) signaling pathway, the NOD-like receptor protein 3 (NLRP3) signaling pathway, the toll-like receptors (TLRs) signaling pathway, the Wnt signaling pathway, Notch signaling pathway, hypoxia-inducible factor 1-alpha and vascular endothelial growth factor (HIF-1alpha/VEGF) signaling pathway, programmed death factor 1 and ligand 1 (PD-1/PD-L1) signaling pathway and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. In recent years, the studies on the treatment of RA with external treatment of traditional Chinese medicine based on the above signaling pathways have gradually increased and deepened. In this paper, we reviewed the relevant data and reports to explain the relationship between the above pathways and the pathogenesis and prevention of RA and summarized the research results on the modulation of the above signaling pathways by external treatment of traditional Chinese medicine to prevent and treat rheumatoid joints with a view to providing a scientific basis for the clinical selection of this treatment.
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Study on optimization of preparation method of frozen section of skeletal muscle tissue

zhangjieyun, liangxinhua

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Objective To explore the optimal frozen section method for fresh and fixed skeletal muscle tissue, and to lay an experimental foundation for the rapid diagnosis of skeletal muscle diseases and the study of the pathogenesis of skeletal muscle diseases.Methods The tibialis anterior muscle of C57BL/6J mice was extracted, and the fresh tissue was treated by liquid nitrogen direct quick freezing, embedding agent combined with liquid nitrogen quick freezing, and foreign body alkane combined with liquid nitrogen quick freezing. The fixed tissues were pre-treated by direct embedding, embedding agent combined with liquid nitrogen quick freezing. After making frozen sections, HE staining was performed, and TISSUEFAX200 was used for scanning. The cross-sectional area of ice crystals and muscle fibers was calculated to evaluate the effect of different pre-treatment methods. Results The morphology of muscle fiber bundles disappeared and a large number of ice crystal vacuoles were observed in fresh tissues after liquid nitrogen direct freezing and foreign body alkane combined with liquid nitrogen freezing. The embedding agent combined with liquid nitrogen quick freezing ( the surface of the tissue is not in direct contact with liquid nitrogen ) shows that the muscle fiber bundle is complete and dense, and there is no ice crystal. It is a pre-treatment method suitable for fresh tissue. Fixed tissue directly embedded is not easy to slice ; after embedding agent combined with liquid nitrogen quick freezing treatment, the muscle fiber bundle was complete and there was no ice crystal.Conclusions The muscle fiber bundles of fresh and fixed tissues treated with embedding agent combined with liquid nitrogen quick freezing are complete and free of ice crystals, which is convenient for further experiments such as immunohistochemistry and immunofluorescence, and is conducive to the accurate and rapid diagnosis of skeletal muscle diseases and the exploration of the pathogenesis of skeletal muscle diseases.
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WEffect of Fuling Xingren Gancao granule on Polarization Phenotype Transformation of Atherosclerotic Macrophages

pengchaojie, wuhong

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Objective To explore the regulatory effect of Fuling Xingren Gancao granule (FXG) on macrophage polarization in atherosclerosis (AS) mice. Methods ApoE -/- mice were used to construct an AS model, while RAW264.7 macrophages were used to construct a polarized model. Oil red O and hematoxylin eosin staining (HE) were used to observe the total area of aortic plaques and the degree of aortic stenosis. PCR and Western blot were used to detect the expression levels, signal transduction, and transcription of M1 polarization factor inducible nitric oxide synthase (iNOS), monocyte chemotactic protein 1 (CCL2), M2 polarization factor arginase-1 (Arg-1), chitinase like protein 3 (YM1), and mannose receptor 1 (Mrcl or CD206) in vitro and in vivo. The phosphorylation level of signal transducer and activator of transcription (STAT3). Result: FXG significantly reduced the total area of aortic plaques in ApoE -/- mice, decreased the expression levels of M1 macrophage polarization factors iNOS and CCL2, and increased the expression levels of M2 macrophage polarization factors Arg-1 and YM1 (P<0.05). The phosphorylation levels of STAT3 in the model group mice and M1 macrophages decreased, but were upregulated after FXG intervention (P<0.05). The P-STAT3 inhibitor Stattic partially eliminated the regulatory effect of FXG on iNOS and Arg-1 (P<0.05).Result FXG significantly reduced the total area of aortic plaques in ApoE -/- mice, decreased the expression levels of M1 macrophage polarization factors iNOS and CCL2, and increased the expression levels of M2 macrophage polarization factors Arg-1 and YM1 (P<0.05). The phosphorylation levels of STAT3 in the model group mice and M1 macrophages decreased, but were upregulated after FXG intervention (P<0.05). The P-STAT3 inhibitor Stattic partially eliminated the regulatory effect of FXG on iNOS and Arg-1 (P<0.05).Conclusion FXG has an inhibitory effect on the progression of AS, which is related to the targeted regulation of macrophage polarization by STAT3.
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Research progress of nuclear factor E2-related factor 2 in oxidative stress after spinal cord injury

xihuilin, wujiajun, wangqingyan, baixinyue, hemengze, yangyanling

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Spinal cord injury (SCI) is a central nervous system disease with high morbidity, high disability and high mortality. A series of pathological and physiological changes after SCI, such as oxidative stress, will promote the further deterioration of the microenvironment at the site of injury, resulting in impaired neurological function. Nuclear factor E2-related factor (Nrf2) is a factor highly correlated with oxidative stress, so targeting the regulation of Nrf2 and alleviating oxidative stress may be one of the treatments for SCI. This article discusses the application of Nrf2 in post-SCI oxidative stress based on the occurrence of oxidative stress after SCI and the relationship between oxidative stress and Nrf2, and summarizes the strategies for targeting and regulating Nrf2, such as genes, non-coding RNAs and drugs, in the hope of providing new ideas for targeted therapy of SCI.
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Advances in the pathogenesis of abnormal copper metabolism disorder in Parkinson's disease

YANG Huijie, LAN Rui, WANG Manman, Wang Weiwei, LI Hongyu, TANG Chen, LIU Shuang, YANG Jieli, SHEN Xiaoming

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Parkinson's disease is a neurodegenerative disease associated with abnormal copper metabolism in the brain,which leads to misfolding and aggregation of α-synuclein-copper complexes, which is an important pathological sign of Parkinson's disease. Copper metabolism refers to the cellular metabolic process involving copper ions, which is closely related to the pathogenesis of α-synuclein aggregation, dopamine metabolism, mitochondrial dysfunction, oxidative stress and iron death in Parkinson's disease. In this review, we describe the molecular metabolic mechanism of copper toxicity by studying the pathological role of copper metabolism in Parkinson's disease，in order to provide evidence and help for further improvement of the mechanism of action and drug development.
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Study on the changes of intestinal permeability secondary to pig’s firearm penetrating wound of abdominal intestine in cold environment at high altitudes

QU Jinquan, LI Jiajia, LU Hongnan, YANG Xingyue, SUN Jiu, LIANG Feixing, WANG Yan, LIU Jiangwei

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Objective To observe the changes and possible mechanism of intestinal permeability in pigs without direct injury after abdominal intestinal firearm penetrating injury in plateau cold environment. Methods 55 experimental pigs were divided into two groups: plateau cold group (HC) and plain normal temperature group (LN). According to the observation time, each group was divided into five experimental subgroups: 0h, 2h, 4h, 8h and 24h. There were 6 pigs in each subgroup in HC group and 5 pigs in each subgroup in LN experimental group. According to the time point of euthanasia, intestinal tissues were taken and the levels of inflammatory factors TNF- α, IL6 and IL-10 in intestinal homogenate and the concentrations of intestinal permeability related proteins DAO and D-lactic acid in blood were detected by ELISA method. The intestinal tissues of experimental pigs were taken at LN-0h, LN-8h and HC-8h time points, and the intestinal pathological changes were observed and scored after HE staining. The contents of Occludin, ZO-1, Claudin-3 proteins related to intestinal permeability and TLR4, NF- κ B and MLCK proteins related to intestinal permeability were detected by WB method to explore the effect of cold environment at high altitude on secondary intestinal permeability changes after injury and its possible mechanism.
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Strain and gender differences in the behavioral models of chronic fatigue syndrome induced by Polyinosinic-polycytidylic acid and swimming in mice

TIAN Jiao, SU rui, WANG tingting, PEI Hailuan, WANG Jing

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Objective To compare the strain and gender differences between the ICR and C57BL/6J mice in the chronic fatigue syndrome model, and provide experimental evidence for the selection of model animals for chronic fatigue syndrome. Methods Mice were injected intraperitoneally with polyinosinic-polycytidylic acid (poly I:C) every three days and swam daily. The modeling was performed for 15 consecutive days, during which the weight and food intake of the mice were measured, and fatigue scores were assessed. After the modeling was completed, behavioral tests were carried out, including exhaustive swimming, tail suspension, mechanical pain threshold, and elevated plus maze. Results Compared with the control group, the exhaustion time of both strains of model mice in exhaustive swimming significantly decreased, the immobility time in tail suspension significantly increased, the mechanical pain threshold significantly decreased, the open arm entry time and frequency of the elevated plus maze in model male mice significantly decreased. The weight of model male C57BL/6J mice significantly decreased. The weight of model female ICR mice increased after the significant reduction. The exhaustion time of control C57BL/6J mice was significantly lower than that of control ICR mice. The immobility time of model C57BL/6J mice was significantly higher than that of model ICR mice. Conclusion There are differences between the two strains of mice in weight, fatigue level, and depression. Among the same strain of mice, there are differences between males and females, and the anxiety level of males is higher than that of females.
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The study of the mechanism by which EFHD2 regulates the NOX4/ROS pathway to initiate glycometabolism reprogramming and promote breast cancer progression

Xu ming fei, Zhang Yi, Kang ru yi, Liu chao yue

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Objective To investigate the mechanism of EFHD2 affecting the occurrence and progression of breast cancer based on NOX4/ROS signaling pathway. Methods The cells were divided into NC-shRNA group and EFHD2-shRNA group. The lentiviral vector silencing EFHD2 expression and its control vector were constructed and transfected MDA-MB-23 and MCF-7 breast cancer cells. The transfection efficiency was verified by qRT-PCR.CCK8 assay was used to detect cell proliferation activity. Plate cloning assay was used to detect cell colony formation ability. Scratch test was used to detect cell migration. Transwell assay was used to detect cell invasion. Flow cytometry was used to detect apoptosis and ROS levels. qRT-PCR was used to detect the mRNA expression of GLUT1, PDK1, PFK1, PKM2, PDH, and LDH. Western blot was used to detect the expression of Cleaved caspase-3, MMP-2 and NOX4 proteins. Results Compared with NC-shRNA group, EFHD2 expression was significantly decreased in EFHD2-SHRNA group, and cell survival and colony formation ability were weakened. The apoptosis rate and the expression of pro-apoptotic protein Cleaved caspase-3 increased. The cell migration distance was shortened, the number of cell invasion and the expression of MMP-2, which promotes migration and invasion, were decreased. The levels of lactic acid and GLUT1, PDK1, PFK1, PKM2 and LDH decreased, while the levels of ATP and PDH increased. Streaming results showed that ROS levels were reduced and NOX4 protein was down-regulated after silencing EFHD2. Conclusion EFHD2 can inhibit ROS production by regulating NOX4/ROS signaling pathway, cause lactic acid and glucose accumulation, promote apoptosis of breast cancer cells, and inhibit cell proliferation, migration and invasion.
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The Current Situation and Suggestions of Research on Laboratory Animal Welfare and Ethics in China

lihuiping, DENG Shaochang, YANG Jingchun, WEN Jingyin, ZHAO Weibo

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Objective? Exploring the current research status, research context, evolution process, and future research directions of laboratory animal welfare and ethics in China. Methods? Using literature related to laboratory animal welfare and ethics collected from source journals in the China National Knowledge Infrastructure (CNKI) database from 2001 to 2023 as the data source, a combination of qualitative description through literature review and CiteSpace visual bibliometric analysis was used to summarize the achievements, hot topics, and directions of laboratory animal welfare and ethics research. Results? Research shows that the overall popularity of research on the laboratory animal welfare and ethics in China is on the rise. The main research institutions include the systematic units of the Chinese Academy of Medical Sciences, the Nanjing General Hospital of the Nanjing Military Region, and the National Institutes for Food and Drug Control. The hot research topics in this field include the basic theories of laboratory animal welfare and ethics, legislation on laboratory animal welfare and ethics, technologies for laboratory animal welfare and ethics, censorship of laboratory animal welfare and ethics, and teaching research on laboratory animal welfare and ethics. In response to ethical issues arising from emerging interdisciplinary fields, continuous innovation is being made in research content. Conclusions? Suggestions have been put forward from the perspectives of legal system, censorship mechanism, education and training, and innovative research of laboratory animals welfare and ethics, with the purpose to provide reference and guidance for welfare and ethics work of laboratory animals.
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Evaluation of a nude mouse model of endometriosis fibrosis by the human xenograft subcutaneous transplantation method

zhouxue, wanguiping, zhangzhenzhen

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Objective To assess the feasibility of establishing a nude mouse model of endometriosis fibrosis of human origin and to determine whether endometrial mesenchymal stem cells are involved in inducing endometriosis fibrosis. Methods Four endometrial tissue specimens were collected and transplanted 1∶3 into 12 BABL/c nude mice by subcutaneous injection. The morphology and volume of the lesions were recorded, and the nude mice were executed on the 15th day after transplantation to observe the morphology of the lesions and their adhesion to the periphery of the abdominal wall, and HE staining was used to judge the results of modelling, Masson staining was used to assess the extent of fibrosis, and immunofluorescence was used to track the role of endometrial mesenchymal stem cells in the fibrotic process of endometriosis. Results The volume of ectopic lesions in nude mice increased over time, showing restricted, vesicular-like changes, with tight adhesion to the abdominal wall, endometrioid glands and collagen fiber deposition were seen microscopically, the success rate of modelling was 83.4%, and the collagen fiber volume fraction of the lesions was significantly higher after modelling (P<0.01), confocal imaging suggested that endometrial mesenchymal stem cells (SUSD2+) could differentiate into myofibroblasts (α-SMA+) in vivo. Conclusion The nude mice model established by the method of human allogeneic subcutaneous injection transplantation is consistent with the lesion characteristics of endometriosis fibrosis, which is simple and feasible, in addition to the in vivo observation that endometrial mesenchymal stem cells are involved in inducing the formation of endometriosis fibrosis, and it provides a better model reference for the further investigation of its pathogenesis.
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Research progress of the mechanism of medial prefrontal cortex layered structure in general anesthesia

lijia, yushouyang, luotianyuan, yutian

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The medial prefrontal cortex engages in various higher brain functions, including attention, transition of consciousness, and information integration, and becomes the central structure of action of general anesthetics. In addition, it is considered both the end point of the bottom-up model of layer-by-layer transmission of information and the starting point of the top-down modulation and integration of information. Classically, the medial prefrontal cortex can be divided into six layers in the vertical direction, with rich neural connections and different functions between layers. In this review, it describes laminar flow structures in the medial prefrontal cortex. Then, the review discusses that the general anesthetics change consciousness via cortical laminar flow structures. To this aim, the article reviews the available evidence in current studies from rodents, primates, and human studies that highlight the role of the medial prefrontal cortex in anesthesia.
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Transcriptome analysis of ethanol extract of Akebia trifoliate (Thunb.) Koidz on cell proliferation inhibitory functions and promote apoptosis in human liver cells

Wang Chunling, WenXiaodong

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Objective To investigate the effects and molecular mechanism of EEATK on proliferation, apoptosis of human hepatocellular carcinoma Hep3B cells and Huh-7 cells and to explore its underlying mechanism . Methods Human hepatoma cells Hep3B cells and Huh-7 cells cultured in vitro, blank control group, Sorafenib group (5uM), EEATK groups (0.10 mg/mL、0.15 mg/mL、0.20 mg/mL、0.3 mg/mL)were set and given corresponding drug intervention, respectively. CCK-8 assay was used to measure the viability of different interventions on the proliferation of Hep3B cells and Huh-7 cells, to screen the optimal action concentration for subsequent experiments. Hep3B cells and Huh-7 cells were divided into the blank control group and EEATK group(0.15mg/mL) treated with for 24 hours,EdU staining assay, colony formation assay were used to explore the effect of EEATK on the proliferation , the apoptosis rate,were detected by Annexin V-FITC/PI double staining assay.The transcriptome sequencing(RNA-seq)technology was used to analyze differentially expressed genes (DEGs) related to cell proliferation, apoptosis on Hep3B cells in blank control group and EEATK group(0.15mg/mL)of Hep3B cells, DEGs were analyzed for Gene Ontology(GO) function and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment. CTD (Comparative Toxicogenomics Database) was used to validate the expression of key proteins related to cell proliferation, apoptosis. Results Compared with the blank control group, different concentration of EEATK could significantly inhibit the activity of Hep3B and Huh-7 cells (P <0. 01) ; After treated with 0.15 mg/mL EEATK, the EdU positive cell rate and cell clone formation rate were significantly decreased (all P <0. 01), at the same time ,the apoptotic rates of EEATK group were significantly increased(P < 0.01). Compared with the blank control group, The transcriptome sequencing of Hep3B cells showed that EEATK induced significant changes in the expression of 1577 DEGs (P <0. 01) , of which 942 were up-regulated and 635 were down-regulated. GO functional enrichment analysis revealed that DEGs were mainly enriched in cholesterol synthesis, inflammation, and extracellular matrix. KEGG pathway analysis showed that EEATK may mainly pass through TFG- β Signal pathways and NF-κB signaling pathway plays an anti-tumor role.DEGs enrichment analysis showed that EEATK can significantly downregulate the expression of apoptosis related genes such as BIRC5 and CDK1 (P<0.01), the expression of CDKN1A and EGLN3 were significantly upregulated (P<0.01), At the same time, EEATK can cause significantly downregulation of cell proliferation related genes such as FAM83D and Ki-67 (P<0.01), mean while, the expression of MYC and FoxC1 were significantly upregulated (P<0. 01).
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Research progress on the improvement effect of Morinda officinalis oligosaccharides on depression

HE Mengjie, DAI Xueling, HUO Qing, LI Xin

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Depression is a psychosomatic disorder. In recent years, the rising incidence of depression around the world, bringing a heavy economic burden on society. The active substances extracted from the Chinese herb Morinda officinalis can soothe depression and calm the mind, tonify the kidney and benefit the intellect, and can also regulate monoamine neurotransmitter levels, enhance neuroplasticity, regulate the HPA axis, and regulate cytokine levels in antidepressant, as well as improve cognitive disorders in patients to a certain extent. Based on the hypothesis of the pathogenesis of depression, this paper summarises the role of Morinda officinalis oligosaccharides in antidepressant, and provides theoretical basis for the research and development of Morinda officinalis oligosaccharides antidepressant drugs.
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Research on potential dominant diseases of Traditional Chinese Medicine in the treatment of geriatric diseases based on bibliometrics

ZUO Yiming, SHI Guohua, LV Shichao, LIU Xuezheng

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Objective In order to effectively use the characteristics and benefits of traditional Chinese medicine in clinical therapy, bibliometric approaches are being used to investigate the Dominant diseases of traditional Chinese medicine in the treatment of Geriatric diseases. Methods The clinical research literature of traditional Chinese medicine in the treatment of Geriatric diseases in the past 10 years was retrieved from CNKI, Wanfang, VIP and CBM. The research trend and clinical efficiency of each disease were statistically analyzed to determine the potential dominant diseases of TCM treatment. Results A total of 22,859 articles were collected, with 3768 being included in the research. According to the International Statistical Classification of Diseases and Related Health Problems (ICD-11) of the World Health Organization, the diseases were classified into 17 categories and 149 diseases, primarily affecting the circulatory system, the skeletal musculoskeletal system or connective tissue system, and the digestive system. Conclusion Traditional Chinese medicine clinical trials on the treatment of geriatric illnesses cover a diverse spectrum of diseases, although the distribution is unequal. Potential dominant illnesses are eventually identified to include osteoporosis, constipation, and hypertension, with heart failure, stroke, coronary heart disease, diabetes and its complications, and insomnia being potential sub-dominant diseases.
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Exploring the effect and mechanism of action of secukinumab on calcific aortic valve disease based on IL-23/Th17 inflammatory pathway

lixinxin, zhangning, fengguangling, lanzhenzhen, guojiao, liuxincan

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Objective To observe whether the IL-23/Th17 inflammatory pathway is involved in the development of calcific aortic valve disease, and whether secukinumab can delay the progression of calcific aortic valve disease by inhibiting the IL-23/Th17 inflammatory pathway. Methods Forty-seven mice were divided into a blank control group, a model group, and a secukinumab group according to the random number table method. The blank control group was fed with normal chow, the model group and the secukinumab group were fed with pro-calcification chow for 16 weeks to establish a calcific aortic valve disease model, and the peak flow velocity changes of the aortic valves were detected under Doppler ultrasonography in all the mice after 4 weeks of intervention by secukinumab on the secukinumab group; relevant indexes were determined by HE staining, Von kossa staining, immunohistochemical staining, ELISA, and PCR. PCR were used to determine the relevant indexes. Results Compared with the model group, 4 weeks of drug intervention in the secukinumab group significantly reduced peak flow velocity ( P<0.05) and serum IL-6 levels ( P<0.05) in the aortic valve; compared with the secukinumab group, the leaflet thickness in the model group was significantly increased and there were more calcium deposits; Immunohistochemical results showed that macrophages ( P<0.05), IL-17A ( P<0.05), and IL-23 ( P>0.05) infiltration in the valve leaflets were reduced in the secukinumab group compared with the model group.PCR results suggested that the expression of STAT3, BMP-2 and α-SMA mRNA were significantly reduced in the secukinumab group compared with the model group ( P<0.05).Conclusion The IL-23/Th17 inflammatory pathway is involved in the pathogenesis of calcific aortic valve disease, and the inflammation, fibrosis, osteogenic differentiation, and calcification of mouse valves were alleviated after the intervention of secukinumab, which may delay the disease progression by inhibiting the IL-23/Th17 inflammatory pathway.
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Research development on the interaction between neuroimmunity and inflammation in the treatment of depressive disorder by traditional Chinese medicine

GUO Dong-jing, PAN Wen-chao, WANG Si-jia, MAO Qian-cheng, ZHAGN Hong-xiu, MA Ke

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As a common mood disorder, depression has a complex pathogenesis. In recent years, with the accelerated pace of people’s lives, the prevalence rate has increased year by year, attracting people extensive attention. External stress and inflammation jointly damage blood vessel and brain functions, induce immune disorders, cause microglia activation and increase the expression of pro-inflammatory cytokines and their receptors, inflammatory reaction continues to occur, which affects the normal metabolism and metabolism of the neurotransmitter system. Abnormal functioning of molecular pathways leads to mutations in brain cells and nerves, and the further formation of the immune-inflammation-neuron cycle path becomes an important mechanism in the occurrence and development of depression. a large number of studies have shown that Traditional Chinese medicine treatment can improve depression symptoms by restoring neuroimmune inflammation homeostasis. From the perspective of neuroimmune inflammation, this article explains its close connection with depression and its pathogenesis, and reviews the role of various traditional Chinese medicine antidepressant therapies in improving and treating depression by participating in the regulation of neuroimmune inflammation to provide a new angle of view for the precise treatment of depression and the development of immune-targeted drugs.
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Research progress on the mechanism of D-galactose-induced brain aging model: A review

ZHU Ziyue, JIN Meiling, XU Xiangyue, LIU Qing, ZHU Jiaxue, FU Mingjun, LEI Xia, ZHANG Ning

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As a normal physiological substance, D-galactose (D-gal) can induce a process similar to natural brain aging in vivo and in vitro under excessive intake, so it is widely used to induce brain aging model in China. The model of brain failure induced by D-galactose has the advantages of short time, low cost and significant effect, but its inducting mechanism is complex and diverse, and the relationships between the mechanisms are not clear, which leads to the limitation of the practical application of the model. This article reviews the in vivo metabolism of D-galactose and the various mechanisms of inducting brain aging, as well as the links between the mechanisms, in order to provide reference for the application and development of this model and the in-depth study of brain aging.
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Role of NLRP3-pyroptosis in experimental sepsis-induced lung injury

LI Yuting, NIU Chunyu, ZHAO Zigang

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Nod-like receptor pyrin domain-associated protein 3 (NLRP3) -mediated pyroptosis of pulmonary parenchymal and immune cells plays a key role in the pathogenesis of acute lung injury (ALI) during sepsis, and NF-?B, JAK2/STAT3, MAPK signaling pathways are involved in NLRP3-mediated pyroptosis. Targeting NLRP3-pyroptosis and its related signaling pathway, the pharmacological intervention of Physalin B, Schisandrin, 4-hydroxynonenal, Baicalin, GYY4137, and Erythropoietin, the physiatrics of acupuncture at Zusanli and Feishu, and the NLRP3 specific inhibitors such as Ergolide, play a good anti-septic effect on ALI. This article reviews the role and mechanism of NLRP3-pyroptosis in sepsis-induced ALI, as well as the experimental progress of targeting NLRP3-pyroptosis in the prevention and treatment of sepsis-induced ALI, hoping to take NLRP3-pyroptosis as the entry point to provide thinking for the formation of a new prevention and treatment strategy for sepsis-induced ALI.
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Effect of folic acid on the expression of Flotillin-1 and Aβ in the brain of mice with AD inflammation

Zewei MA, Li HUANG, Yunqin ZHENG, Meilin ZHANG, Huan LIU

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【Objective】To observe the effects of folic acid (FA) supplementation on the expression of Flotillin-1 and β-amyloid protein (Aβ) metabolism-related proteins in the brains of inflammation-stimulated AD mice.【Methods】Twenty-seven 6-month-old male APP/PS1 mice were randomly divided into AD, AD+LPS, and AD+LPS+FA groups, with 9 mice in each group. 9 same-month-old C57BL/6J male mice were used as the wild control group (Control). The AD+LPS+FA group was given folic acid supplemented feed (8 mg/kg) for three months of intervention, while the other three groups were fed normal feed. Lipopolysaccharide solution (LPS, 250 μg/kg/day) was injected intraperitoneally into mice in the AD+LPS group and AD+LPS+FA group one week before the end of the experiment, and saline was injected into the remaining two groups. The serum inflammatory factors TNF-α and IL-6 levels and brain tissue Aβ1-40 and Aβ1-42 levels of mice in each group were detected by ELISA, Flotillin-1 protein expression in brain tissue was detected by Western blot, and immunofluorescence double-labeling was used to detect the co-expression of Flotillin-1 in the cortical region of the brain and Aβ1-42/APP/PS1/ BACE1 co-expression. 【Results】 After ANOVA analysis, compared with the Control group, mice in the AD group had elevated serum TNF-α and IL-6 levels (P < 0.05), elevated levels of Aβ1-40 and Aβ1-42 (P < 0.05), increased expression of Flotillin-1 protein (P < 0.05), and increased co-expression of Flotillin-1 and Aβ1-42/APP/PS1/BACE1 in the cortical region of brain tissue (P < 0.05); Compared with the AD group, mice in the AD+LPS group had further increases in serum inflammatory factors and Aβ levels in the brain (P < 0.05), and increased expression of double-labeled positive proteins in the cortical region of brain tissue for Flotillin-1 and Aβ1-42/APP/BACE1 (P < 0.05) ; Compared with the AD+LPS group, mice in the AD+LPS+FA group had lower in vivo inflammation levels and Aβ content in the brain (P < 0.05), lower brain tissue Flotillin-1 protein expression (P < 0.05), and lower co-expression of Flotillin-1 with Aβ1-42/APP/PS1/BACE1 proteins in the cortical region of brain tissue (P < 0.05). 【Conclusion】 Folic acid supplementation may reduce Flotillin-1 protein expression and Aβ deposition in the brain of AD inflammatory mice.
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Practice and reflection on combining teaching and training to promote the standardization of animal experiments for medical postgraduates

CHEN Qin, LI Jianmin, GAO Yun, QIU Chen

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Standardized animal experiments are the basic prerequisite and important guarantee for eliminating potential biosafety risk factors and obtaining accurate medical research results. Professional training for graduate students who conduct animal experiments and strengthening publicity and education on animal experiment management regulations can help establish a sense of responsibility for standardizing animal experiments among medical graduate students. The innovative teaching-training model will fundamentally help to ensure the smooth implementation of animal experiments, and truly promote the healthy development of laboratory animal work in universities.
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The effect of Angelica sinensis polysaccharides on Th17/Treg cell balance and expression of related inflammatory factor proteins in bone marrow of mice with aplastic anemia

chenxiubao, xufei, yinxi, chenweida, cuixing, liuxiujuan, chenzetao

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Objective：To investigate the effect of Angelica sinensis polysaccharides on the balance of Th17/Treg cells and the expression of related inflammatory factor proteins in bone marrow of mice with aplastic anemia.Methods 50 male BALB/c mice were randomly divided into normal group,model group,cyclosporineA group,APS low and high dose groups.A mouse model of aplastic anemia was prepared by combining radiation irradiation with allogeneic lymphocyte infusion.After modeling,the mice were orally administered for 28 days.The general condition,weight changes,and spleen index of the mice were observed.Blood samples were taken to test the changes in basic hematological parameters of peripheral blood,and HE staining was used to evaluate the pathological changes in mouse bone marrow tissue;ELSIA method for detecting bone marrow TGF-β1.The expression levels of proteins such as IL-10,IL-17A,and IL-6;FCM detection of bone marrow Th17,Treg cell ratio,and Th17/Treg cell balance changes.Results Compared with the normal group,the model group mice showed clumsiness in activity,lethargy,pale eyelids,lips,and ears,and reduced food and water intake;Both body weight and spleen index significantly decreased;The peripheral blood red blood cell,white blood cell,platelet count,and hemoglobin concentration were significantly reduced;Disordered bone marrow tissue structure and significantly reduced proliferation of hematopoietic tissue;The expression of IL-17A and IL-6 proteins is significantly upregulated,and TGF-β1.The expression of IL-10 protein was significantly downregulated;The proportion of Th17 cells in bone marrow significantly increased,the proportion of Treg cells significantly decreased,and the ratio of Th17/Treg cells significantly increased(P<0.01).Compared with the model group,the general condition of the low and high dose APS groups improved,with an increase in food and water intake,a significant increase in body weight and spleen index,a significant increase in peripheral blood red blood cell,white blood cell,platelet count,and hemoglobin concentration(P<0.05),an improvement in bone marrow tissue structure,and an increase in hematopoietic tissue proliferation (P<0.05);The expression of IL-17A and IL-6 proteins is significantly downregulated,and TGF-β1. The expression of IL-10 protein was significantly upregulated(P<0.05 or P<0.01);The proportion of Th17 cells in bone marrow significantly decreased,the proportion of Treg cells significantly increased,and the ratio of Th17/Treg cells significantly decreased (P<0.01).Conclusion Angelica sinensis polysaccharides can improve bone marrow hematopoietic function in AA mice,and its mechanism may be achieved by regulating Th17/Treg cell balance.
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Analysis of ankylosing spondylitis’ animal model based on clinical characteristics of traditional Chinese and western medicine

Zhang Fangzhi, Miao Furui, Fan Yushan, He Yujun

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Ankylosing spondylitis is an autoimmune disease with sacroiliac arthritis and spinal arthritis as the main manifestations. The disease mainly occurs in young men, with a high disability rate and a serious threat to the life and health of patients. Biological agents are expensive, and there are many adverse reactions of hormones, non-steroidal anti-inflammatory drugs and anti-rheumatic drugs. Traditional Chinese medicine can regulate immunity and anti-inflammatory, and has a good clinical effect. In order to further study the pathogenesis of ankylosing spondylitis and the development and screening of therapeutic drugs, this paper summarizes the modeling method and mechanism of the existing animal model of ankylosing spondylitis, and analyse advantages and disadvantages of the model. To evaluate the coincidence of Clinical Characteristics of Chinese Medicine and Western Medicine,the paper compares the characteristics of Chinese and Western medical syndromes of the animal model. To build an animal model of ankylosing spondylitis with a higher degree of coincidence between traditional Chinese and western medicine is the key to innovative research of traditional Chinese medicine for treating ankylosing spondylitis. In order to lay a foundation for the research of traditional Chinese medicine for ankylosing spondylitis，this paper analyzes the degree of coincidence between Clinical Characteristics of Chinese Medicine and Western Medicine of animal model.
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Advances in bee venom research: novel drug candidate library for the treatment of inflammatory skin diseases

WU Qiang, Yang Yue, LI Peng, GU Jiajie, DONG Degang, YI Jun

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Inflammatory skin diseases (ISD) are characterized by persistent inflammatory infiltration and lingering and intractable skin lesions. At present, corticosteroids are the main drugs used in the treatment of ISD. However, due to the characteristics of recurrent and intractable ISD, long-term use of these hormone drugs may cause serious side effects to patients. In recent years, more and more studies have confirmed that bee venom (BV) has significant anti-inflammatory, anti-apoptosis, anti-fibrosis, antibacterial and other effects, so as to effectively treat ISD. In this paper, the main active components of BV are reviewed and its anti-inflammatory mechanism, and the latest status of BV in acne, atopic dermatitis, psoriasis, urticaria and systemic lupus erythematosus is discussed, which provides reference for the basic research and clinical treatment of ISD.
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Effect and Mechanism of Qishishenshu Capsule on Renal Fibrosis in Mouse Early Diabetic Nephropathy

LAO Xiao-qing, CHEN Chen, ZHANG Hong-min, YANG Xiu, SHI Jiang-shan, SU Hong-wei, SHEN Hong-ping, WANG Li, YOU Man-rui, LI Xiao-bin, ZHAO Chang-ying

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【Abstract】Objective To investigate the therapeutic effect and underlying mechanism of Qishishenshu capsule on renal fibrosis in mice with early diabetic nephropathy (DN). Methods DN mouse model was established by multiple injection of streptozotocin (STZ). The mice were randomly divided into the normal group (NC), model group (DN) and Qishi group (QS)(0.9 g/kg/d), 8 mice in each group, gavaged continuously for 4 weeks, and fasting blood glucose (FBG) was measured weekly. 4 weeks later, urinary microalbumin creatinine ratio (UACR), serum creatinine (SCr) and blood urea nitrogen (BUN) were measured. Hematoxylin-eosin staining (HE staining), periodic acid-Schiff staining (PAS staining) and Sirius red staining were used to analyze renal pathological changes. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect the Message RNA (mRNA) levels of fibronectin (FN), collagen type I alpha 1 (Col1a1), and α-smooth muscle actin (α-SMA). Immunohistochemistry (IHC) and Western blot (WB) were performed to detect FN, collagen type I (Collagen I), collagen type III (Collagen III), α-SMA, Podocin, Nephrin and transforming growth factor-β1/SMAD family member2/3 (TGF-β1/Smad2/3) pathway-related proteins. Results Compared with the NC group, mice in the DN group showed significantly higher levels of FBG and UACR (P<0.001). Mesangial hyperplasia, basement membrane thickening and collagen deposition occurred in renal tissue. The mRNA levels of FN, Col1a1 and α-SMA were increased (P<0.05). Protein levels of Podocin and Nephrin were decreased (P<0.05). The levels of FN, Collagen I, Collagen III, α-SMA and TGF-β1/Smad2/3 pathway protein were increased (P<0.05). Compared with the DN group, the level of UACR in QS group was decreased (P<0.05), and renal pathological injury was alleviated. The mRNA levels of FN, Collagen1 and α-SMA were attenuated (P<0.05). The protein levels of Podocin and Nephrin were elevated (P<0.05). Protein levels of FN, Collagen I, Collagen III, α-SMA and TGF-β1/Smad2/3 pathway protein were also decreased (P<0.05). Conclusions Qishishenshu Capsules improved renal fibrosis in DN mice probably through the inhibition of TGF-β1/Smad2/3 signaling pathway.
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Research progress of diarrhea animal model and its drug therapy

Shi Yucun, Dong Xiaoli, Hou Xiaoying, Yin Kai, Gao Fan, Wu Guotai

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Diarrhea is a common and frequently occurring disease in clinic, and there are many factors causing diarrhea. At present, there are many methods to study the animal model of diarrhea, but there are limited kinds of drugs to treat diarrhea in clinic, some of the drugs available also apply to single-factor diarrhea. Therefore, the establishment and selection of diarrhea animal model is not only an important basis for further study of the pathogenesis of diarrhea, but also an effective means for clinical screening and evaluation of all-round anti-diarrhea drugs. This article reviews the establishment and evaluation of animal models of diarrhea and the progress in the research of therapeutic drugs, in order to provide reference for the animal experimental research and drug prevention and treatment of diarrhea.
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Overexpression of TFB1M inhibits hepatocellular carcinoma cell migration and invasion

Tao Jia, He linli, wang li, ge pengsheng, sai shunhai

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Objective To explore the effect of transcription factor B1,mitochondrial,(TFB1M) on cell survival and migration of hepatocellular carcinoma cells and related molecular mechanisms. Methods TFB1M gene was overexpressed by transfection of eukaryotic recombinant plasmid. Cell survival and cell cycle distribution were detected by CCK-8 and flow cytometry. Cell migration and invasion were assessed by scratch healing and transwell chamber assays. Results The expression of TFB1M was significantly decreased in HCC, and the expression of TFB1M was more lower in female HCC patients than that tin male HCC patients. The mRNA expression of TFB1M was significantly increased after transfection of recombinant plasmid. Moreover, our results showed that overexpression of TFB1M had no effect on the growth and cell cycle distribution of HepG2 and Huh7 cells. However, overexpession of TFB1M remarkably attenuated the migration and invasion activity of HCC cells(all P<0.05). The results from the bioinformatics presented that the low expression of TFB1M might be related to the extracellular matrix assembly gene set in HCC. Conclusions TFB1M regulates the cell metastasis and invasion through influencing extracellular matrix assembly in HCC.
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Research progress on the mechanism of STAT3 in diabetic kidney disease

liuruidi, xujiangyan, xiezhishen, zhangxiaowei, chenjie, zhangzhenqiang

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Signal transducer and activator of transcription 3 (STAT3) is known to modulate expression related to cell transformation, proliferation and survival making it a significant target for cancer therapy. Recent research has also highlighted the crucial involvement of aberrant STAT3 activation in the pathogenesis of diabetic kidney disease (DKD). This paper aims to explore the therapeutic implications of targeting STAT3 in DKD, encompassing its structural characteristics, regulatory mechanisms of activity, pathological activation in DKD, and an overview of current research progress, with the objective of advancing understanding of DKD pathogenesis.
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Research Progress on Macrophage Metabolism and Immune Function in Coronary Heart Disease (CHD) with Phlegm and Blood Stasis Syndrome

CAO Panxia, PENG Zining, LI Jian, WU Hong

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In traditional Chinese medicine, coronary heart disease falls under the categories of "chest impediment and heart pain" and "true heart pain", with lipid metabolism disorder and inflammatory response acting as biochemical manifestations of "phlegm and stasis" throughout the disease. The energy metabolism of macrophages is closely related to their immune function and is an important factor in regulating the metabolic disorder and inflammatory response in coronary heart disease. This article reviews the role of macrophages in the pathophysiology of coronary heart disease, discusses how these metabolic pathways affect the immune response of macrophages, and how they influence the disease through energy metabolism pathways. It delves into the different modes of macrophage energy metabolism, especially the metabolic characteristics and immune regulatory functions of pro-inflammatory M1 and anti-inflammatory M2 macrophages in the syndrome of phlegm and blood stasis in coronary heart disease. This provides theoretical guidance for understanding the pathogenic mechanism of the syndrome of phlegm and blood stasis in coronary heart disease and developing new treatment strategies.
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Research progress on pharmacological mechanism of traditional Chinese medicine in the treatment of cerebral ischemia-reperfusion injury based on regulating AMPK signaling pathway

hechenfei, machiyuan, ranchunlong, chenghaoge, liuxiangzhe

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Cerebral ischemia-reperfusion injury(CIRI)refers to the recovery of blood supply after cerebral ischemia, leading to further damage and dysfunction of brain tissue. At present, modern medicine has made some progress in the prevention and treatment of CIRI, but it still faces some challenges and limitations. Therefore, it is of great clinical value to find effective interventions to prevent and treat CIRI. AMP-activated protein kinase(AMPK)and its downstream proteins are important targets for the treatment of CIRI and play a key role in the regulation of cellular energy homeostasis. Traditional Chinese medicine has the characteristics of multi-target, multi-pathway and multi-effect. It can activate the cascade reaction of AMPK signaling pathway, and treat CIRI by regulating autophagy, oxidative stress, inflammatory response and apoptosis, and has achieved certain results. Therefore, this paper summarizes the structure and mechanism of AMPK-related signaling pathway, elaborates its relationship with CIRI, and systematically summarizes the research status of traditional Chinese medicine regulating AMPK signaling pathway in the prevention and treatment of CIRI, in order to provide new ideas for the prevention and treatment of CIRI in traditional Chinese medicine and the development of new drugs.
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Effects of hypoxic preconditioning on NR2B in mouse hippocampal cell and HT22 cell

Wang Zhigang, Liu Xiaolei, yanlei, Wang Zhiguang, zhang Zhiyong, JIANG shuyuan, yang jing, Shao Guo

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objective: N-methyl-D-aspartate (NMDA) receptor subunit 2B (NR2B) and its phosphorylation are involved in cerebral ischemia/hypoxic neural injury. Hypoxic preconditioning (HPC) serves as an endogenous protective intervention to protect the brain from ischemic/hypoxic injury. This study intends to explore the effect of HPC on NR2B and its two tyrosine sites phosphorylation (1252 and 1336) in hippocampal cells through in vivo and in vitro experiments, and explore its role in HPC neuroprotection.
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Research progress on the mechanism of epigenetics in improving Parkinson"s disease through exercise

Liu Bo, Chen XiangHe, LuPengcheng, YangKang

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PD is a highly complex neurodegenerative disease, and its pathogenesis is influenced by DNA methylation, histone modification, and non-coding RNA. In research on Parkinson"s brain tissue and blood, it has been shown that changes in DNA methylation/histone modification levels, variations in ncRNA and its target genes may trigger or exacerbate the pathogenesis of neurodegeneration, and serve as potential non-invasive biomarkers for Parkinson"s disease. Exercise can improve neurodegenerative diseases caused by aging and PD by reversing epigenetics. This article reviews the pathogenesis of epigenetics in Parkinson"s disease, explores the role of exercise in it, and provides theoretical reference for improving Parkinson"s disease by regulating epigenetics through exercise.
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Inhibition of Boschniakia rossica Boschnaloside on Epithelial-Mesenchymal Transition of Hepatoma SMMC-7721 Cells

JIN Aihua, ZHU Jiebo, QUAN jishu

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Objective To investigate the inhibitory effect of Boschniakia rossica boschnaloside (BRBN) on epithelial-mesenchymal transition (EMT), invasion and migration of human hepatoma SMMC-7721 cells. Methods Transforming growth factor β1 (TGFβ1) was used to induce EMT model of SMMC-7721 cells. The cells were divided into the control group, model group and BRBN group. The cell migration was detected with wound healing test, the cell invasion was observed by transwell chamber assay, and the expressions of E-cadherin, N-cadherin, vimentin, Slug, Twist1, ZEB1 and signal transducers and activators of transcription 3 (STAT3) were determined with the western blot method. Results TGFβ1 resulted in the spindle-shaped changes in SMMC-7721 cells, down-regulated the expression of E-cadherin, and up-regulated the expression of N-cadherin and vimentin, suggesting that SMMC-7721 cells have obtained the mesenchymal phenotype. Compared with the model group, the expression of E-cadherin was significantly increased, expressions of N-cadherin and vimentin were reduced, expressions of Slug, Twist1, ZEB1 as well as STAT3 phosphorylation were also down-regulated in SMMC-7721 cells of BRBN group. Conclusions BRBN could inhibit the invasion and migration of human hepatoma SMMC-7721 cells, possibly by reversing EMT through STAT3 pathway.
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The Relationship Between Insulin Resistance, Hyperandrogenism, and Mitochondrial Oxidative Stress in PCOS

zhaoqi, yangliping, chenping, sunjianhua, mazibo

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Polycystic ovarian syndrome (PCOS) stands as a prevalent reproductive endocrine metabolic disorder within obstetrics and gynaecology. Two pivotal features of PCOS are insulin resistance (IR) and hyperandrogenemia (HA), both instrumental in its pathogenesis. There exists a discernible correlation between PCOS patients and mitochondrial oxidative stress, a relationship heavily influenced by various factors, notably insulin resistance and hyperandrogenemia. This article endeavors to elucidate the interplay between insulin resistance, hyperandrogenemia, and mitochondrial oxidative stress, while also exploring the impacts of insulin resistance and hyperandrogenemia on mitochondrial oxidative stress among PCOS patients.
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Research Progress on the Role of Microglia Efferocytosis in Alzheimer's Disease

Machiyuan, Wangxinzhi, Liuxiangzhe, Hanzhenzhen, Guoming, Lixiao

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Microglia efferocytosis is the process by which microglia phagocytose damaged and dead cells, with anti-inflammatory and pro-damage repair effects. Recent studies have shown that microglia efferocytosis plays a crucial role in the pathogenesis of Alzheimer"s disease (AD) and may be a novel therapeutic target for AD. This paper reviews the relationship between microglia efferocytosis and AD pathogenesis and the potential of efferocytosis-related molecules as therapeutic targets for AD. The aim is to provide new ideas and approaches for the treatment of AD.
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Research Progress on the Intervention of Tryptophan Metabolism in Alzheimer"s Disease

WU Cui, LI Canwei, ZHAO Hairong, YANG Zizhong, GAO Pengfei, ZHAO Yu, ZHANG Chenggui

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Alzheimer"s Disease (AD) is a degenerative neurological disorder that can lead to cognitive decline, mental behavior abnormalities, and reduced ability to daily lives. Tryptophan is an essential amino acid for the human body and is produced by three main metabolic pathways, namely kynuridine, 5-hydroxytryptamine (5-HT) and indole derivatives. Influencing the metabolites of tryptophan can lead to ameliorated neuroinflammation in the brain and significantly improvdcognitive ability, while the occurrence and development of AD.is affected and intervened. In this paper, we reviewed the research literature on tryptophan metabolism intervention in AD in recent 3 years from CNKI, PubMed and other databases.
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Role and mechanism of DNMT1 regulating LSM4 in HCY-induced hepatocyte apoptosis in mice

Xia Tongtong, Ma Fang, Liu Honglin, Zhang Zhenghao, Ding Hanshuang, Hao Yinju, Zhang Huiping, Wu Kai, Jiao Yun, Jiang Yideng, Li Guizhong

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Objective To study the effect of DNA methyltransferase 1 (DNMT1) on sm-like protein-4 (LSM4) in the hepatocytes apoptosis of mice induced by Hcy. Methods 12 ApoE-/- mice were divided into two groups: normal diet (ND, n = 6), high methionine diet (HMD, n = 6). Normal hepatocytes of NCTC1469 were divided into normal group (control, 0 μL/L Hcy), Hcy intervented group (Hcy, 100 μL/L Hcy), transfected NC siRNA control group (si-NC group, 0 μmol/L Hcy), transfected LSM4 siRNA group (si-LSM4 group, 0 μmol/L Hcy), transfected DNMT1 siRNA group (si-DNMT1 group, 0 μmol/L Hcy), transfected NC siRNA in the Hcy intervened group (Hcy＋si-NC group, 100 μmol/L Hcy), transfected LSM4 siRNA in the Hcy intervened group (Hcy＋si-LSM4 group, 100 μmol/L Hcy) and transfected siRNA DNMT1 in the Hcy intervened group (Hcy＋si-DNMT1 group, 100 μmol/L Hcy); Analysis of the expression of LSM4 in various tissues by NCBI database; quantitative real-time PCR (qRT-PCR) and Western blot (WB) were used to detect the difference of LSM4 protein expression in mice tissues (HMD and ND) and hepatocytes (control and Hcy). Western blot was used to detect the protein expression of Bcl2-associated X (Bax) and B-cell lymphoma-2 (Bcl-2). The cell apoptosis rate of control, Hcy, Hcy＋si-NC and Hcy＋si-LSM4 were detected by flow cytometry. MethPrimer online software was used to analyze the CpG islands of LSM4 promoter region. The expression of LSM4 in si-DNMT1 group was detected by qRT-PCR and Western blot. Results The expression of LSM4 in HMD/Hcy group was higher than that in the control group (P＜0.05). Bax protein expression in Hcy＋si-LSM4 was significantly higher than that in the control group (P＜0.05), but Bcl-2 was significantly lower (P＜0.05). The expression of Bax in Hcy＋si-LSM4 was significantly lower than that in Hcy＋si-NC group (P＜0.05), and the level of Bcl-2 was significantly increased. The cell apoptosis rate in Hcy group was higher than that in the control group (P＜0.05). The cell apoptosis rate in Hcy＋si-LSM4 group was lower than that in the Hcy＋si-NC group (P＜0.05). MethPrimer database analysis showed that the promoter region of LSM4 was GC rich and there was one CpG island. Compared with the Hcy+si-NC group, the expression of LSM4 protein in the Hcy+si-DNMT1 group was increased (P＜0.05). Conclusions DNMT1 regulates LSM4 hypomethylation to increase its expression, thereby promoting Hcy-induced apoptosis of mouse hepatocytes.
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Establishment of a multi factor induced hyperuricemic nephropathy rat model and study on the intervention effect of Qiling granules

Zhang Qian, Tu Hai Ye, Zhu Ke Yan, Yu Chen, Cai Yue Qin, Rong Yi Li, Zhang Li Zong, Chen Min Li, Fang Ming Sun

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Objective To establish a rat model of hyperuricemic nephropathy (HN) using a multifactorial induction method of potassium oxazinate combined with adenine and yeast feed, and to observe the intervention effect of Qiling granules (QLG). Methods Fifty-eight SPF grade male SD rats were selected, and ten rats were randomly selected as the normal control group. The remaining rats were induced by multiple factors to establish HN rat models. After 2 weeks of modeling, submandibular blood samples were taken to detect serum UA, CREA, BUN, TG, and TC. Forty HN rats with bleeding clearance UA and body weight close to the mean were selected. They were randomly divided into model control group, QLG low and high dose groups, and positive control group using a stratified randomization method, with ten rats in each group. Each group was given corresponding drugs by gavage daily, and after continuous administration for 4 weeks, submandibular blood samples were taken to detect serum UA, CREA, BUN, TG, and TC. After euthanasia of rats, and liver tissue was taken to detect XOD and ADA activity. Renal tissue was taken for HE and Gomori hexamine silver staining, and the protein expression of GLUT9, OAT1, VCAM-1, and TGF-β in the kidneys was observed using immunohistochemistry and WB methods. Results Compared with the normal control group, the serum levels of UA, CREA, BUN, TC, and TG, as well as liver XOD and ADA activities, were significantly increased in model control Group (P<0.01). The renal tissue of rats shows significant pathological changes. The positive area of urate in renal tubules and the expression of GLUT9, VCAM-1, and TGF-β proteins in the kidneys were significantly increased (P<0.01, P<0.05), while the expression of OAT1 was significantly reduced (P<0.01). Compared with model control group, the serum UA levels, liver XOD, ADA activity, and renal VCAM-1 protein expression of rats in each treatment group were significantly reduced (P<0.01, P<0.05). The serum CREA, BUN levels, and renal TGF-β protein expression of rats in the low-dose QLG group were also significantly reduced (P<0.05, P<0.01). The serum CREA, BUN levels, and renal GLUT9 protein expression of rats in the high-dose QLG group were also significantly reduced (P<0.01, P<0.05), and the urate deposition and renal injury caused by them in each treatment group were reduced to varying degrees, but there was no significant difference (P>0.05). Conclusion A stable HN rat model can be induced by gavage of potassium oxyzinate and adenine in combination with yeast feed to rats; QLG can effectively treat HN by improving UA metabolic disorders, reducing renal inflammation and urate deposition, as well as causing renal damage in HN model rats. Its mechanism of action is related to reducing serum UA, CREA, BUN, TG levels, liver XOD and ADA activities, as well as the expression of GLUT9, OAT1, VCAM-1, and TGF-β proteins in the kidneys.
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Ferroptosis and its role during experimental acute lung injury

guohuanrong, wang jing, zhao zi gang, niu chun yu

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Ferroptosis is a non-apoptotic mode of cell death characterized by iron-dependent lipid peroxides accumulation-induced membrane lipid peroxidation and mitochondrial atrophy, and differed from other programmed cell deaths in morphological and biochemical properties. Ferroptosis is regulated by a variety of metabolic pathways, involved in acute lung injury induced by hemorrhagic shock, ischemia-reperfusion, sepsis, and radiation. This article reviews the main regulatory mechanisms of ferroptosis and its role in the pathogenesis of acute lung injury in various animal models, with the aim of providing new strategies for the prevention and treatment of acute lung injury.
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Study of the mechanism of Honokiol protecting pulmonary microvascular endothelial barrier in LPS induced acute lung injury

Liu Jinxing

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Objective: To study the effect of Honokiol (HKL) on pulmonary microvascular endothelial cells in lipopolysaccharide (LPS) induced acute respiratory distress syndrome (ARDS) and its potential mechanism.Methods: Mouse lung microvascular endothelial cells (PMVEC) were cultured with DMEM+10%FBS in a six-well plate and divided into control group (Con group), Honokiol group (HKL group), Lipopolysaccharide treatment group (LPS group) and LPS+HKL treatment group (HKL+LPS group). The levels of Malondialdehyde and Reactive Oxygen Species in cell lysates were determined by TBARS assay kit and H2DCF-DA, respectively. TUNEL/DAPI double staining was used to detect apoptosis. The cell junction was detected by VE-cadherin/DAPI and Claudin-5/DAPI double staining. Western blotting was used to detect caspase-3, cleaved caspase-3, Sirt3, SOD2, and acetylated SOD2 (Ac-SOD2). 32 mice were randomly divided into control group (Con group), Honokiol group (HKL group), Lipopolysaccharide treatment group (LPS group) and LPS+HKL treatment group (HKL+LPS group). HE staining was used to observe the pathological changes of lung tissue.Results: Honokiol pretreatment could significantly reduce LPS-induced increase of ROS and MDA levels, and decrease LPS-induced increase of SOD2 acetylation and SIRT3 down-regulation. TUNEL and Caspase analysis showed that Honokiol could protect apoptosis of PMECs induced by LPS. VE-cadherin fluorescence staining demonstrate thag Honokiol pretreatment could prevent LPS from damaging cell adhesion. Claudin-5 fluorescence staining show that Honokiol pretreatment could prevent LPS from disrupting tight connections of cells. In animal experiments, HE staining showed that Honokiol significantly inhibited the typical pathological changes of ARDS in the lung tissue of mice in LPS group.Conclusion: HKL can significantly inhibit LPS-induced oxidative stress, apoptosis and cell connection breakdown of pulmonary microvascular endothelial cells, thereby alleviating ARDS symptoms.
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Preliminary studies on the mode of CdCl2 in combination with nicotine on spermatogonial cells

han xue, jiang li, zhang yuan, wu de sheng, huang hai yan, liu jian jun

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Objective：To investigate the combined effect of CdCl2 and nicotine on the cell cycle and apoptosis of mouse spermatogonium(GC-1).Methods：CCK-8 assay was used to detect the inhibition of GC-1 cell proliferation in the CdCl2 groups, the nicotine groups, and the combined groups at 24 hours. Flow cytometry was used to detect the effects of the CdCl2 groups, the nicotine groups, and the combined groups at 24 hours on the cell cycle and apoptosis of GC-1 cells. Results：The inhibitory effect on the proliferation of GC-1 cells was increased with the rise of CdCl2 and nicotine concentrations , the IC50 values of CdCl2 and nicotine alone was 5.409μmol/L, 2814μmol/L. When CdCl2 and nicotine were used in combination, the IC50 value of GC-1 cells was 4.422 μmol/L for 0.175 mmol/L nicotine, 4.532 μmol/L for 0.35 mmol/L nicotine, 3.309 μmol/L for 0.7 mmol/L nicotine, and 2.532 μmol/L for 1.4 mmol/L nicotine ,all of which were lower than that of the groups in which CdCl2 was used alone. The combination of CdCl2 and nicotine had a synergistic effect. At a CdCl2 concentration of 2.5 μmol/L, the IC50 concentration of the combination group was lower than that of nicotine alone. The results showed that percentage of G0/G1 phase cells was decreased in the 2.5 μmol/L CdCl2 group and 1.4 mmol/L nicotine group; compared with the CdCl2 groups and the nicotine groups, percentage of G0/G1 phase cells was decreased in the CdCl2 and nicotine combination groups; percentage of G2/M phase cells was increased in the 2.5 μmol/L CdCl2 group, and compared with the CdCl2 groups and nicotine groups, the cells were blocked in G2/M phase in the combined groups. Combined effects of CdCl2 and nicotine on the cell cycle had a synergistic effect , and the effect of CdCl2 on the synergistic effect was stronger. Proportion of cell apoptosis was increased in 2.5μmol/L CdCl2 group. Compared with the CdCl2 groups and the nicotine groups, percentage of cell apoptosis in the CdCl2 and nicotine combined groups were increased significantly, and the effect of CdCl2 was stronger for the synergistic effect. Conclusion：Combination of CdCl2 and nicotine had a synergistic effect on mouse spermatogonium(GC-1) which could lead to enhanced cytotoxicity, G2/M cell cycle arrests, and cell apoptosis. And CdCl2 was the main effect factor of synergistic effect.
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Molecular mechanism of autophagy mediated by AKT/mTOR pathway involved in exercise rehabilitation of rotator cuff tear-related muscle atrophy

tangjing

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objective: To explore the molecular mechanism of autophagy mediated by protein kinase B(AKT)/ mammalian target protein of rapamycin (mTOR) pathway in the rehabilitation of muscle atrophy associated with rotator cuff tears (RCTs). Methods: Forty male C57 mice were randomly assigned to the following four groups: sham group, RCTs group, RCTs+ exercise group and RCTs+ exercise+rapamycin group, with 10 mice in each group. At the eighth week after grouping, the healing of bone-tendon interface and muscle cell atrophy were analyzed by histology. The mRNA expression levels of muscle atrophy related genes (Atrogin-1, Bnip 3, MuRF-1) in supraspinatus muscle tissue were measured by real-time quantitative reverse transcription polymerase chain reaction. The expression of LC3 and AKT/mTOR signal pathway in supraspinatus muscle tissue of different groups was detected by western blot, and the production of autophagy in each group was analyzed by transmission electron microscope. Results: Compared with sham operation group, the maturity score of bone-tendon interface at supraspinatus tendon anchorage and the cross-sectional area of supraspinatus muscle fibers in RCTs group decreased significantly (P<0.001), and the muscle loss and the expression of Atrogin-1, Bnip 3 and MuRF-1 genes increased significantly (P<0.001). Compared with RCTs group, RCTs+ exercise group significantly increased the bone-tendon interface maturity score and the cross-sectional area of supraspinatus muscle fibers (P<0.01), and decreased the muscle loss and the expression of Atrogin-1, Bnip 3 and MuRF-1 genes (P<0.01). Compared with the sham group, the LC3Ⅰ/LC3Ⅱ and number of autophagy in supraspinatus muscle of RCTs group increased significantly (P<0.001), while the p-Akt/Akt, p-mTOR/mTOR decreased significantly (P<0.01). Compared with RCTs group, the LC3Ⅰ/LC3Ⅱ and number of autophagy in RCTs+ exercise group decreased significantly (P<0.01), and the number of p-Akt/Akt and p-mTOR/mTOR increased significantly (P<0.001). The addition of rapamycin significantly inhibited the improvement of exercise on bone-tendon interface healing, muscle atrophy and loss, AKT/mTOR signaling pathway and autophagy number induced by RCTs. Conclusion: This study confirme the anti-atrophy effect of exercise rehabilitation in RCTs diseases, and its mechanism is related to activating AKT/mTOR signal to inhibit autophagy.
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The overview of traditional Chinese medicine in regulating the mechanism of regulatory cell death in vascular dementia

lihui, Dongxingang, Liweifeng

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Vascular dementia (VD) is a neurodegenerative disease caused by brain injury, the research on the process of its occurrence is not yet in-depth. The regulatory all death (RCD) mechanism is related to the pathological mechanisms of VD, including apoptosis, pyroptosis, autophagy, ferroptosis, and copper mediated cell death proposed in recent years. Therefore, explain the role between the mechanisms of regulatory death and the pathological mechanisms of VD, is of beneficial for the study of VD. The article provides a review of the roles of five mechanisms of RCD in VD, and summarizes the recent research progress in the treatment of VD with traditional Chinese medicine, in order to development of new traditional Chinese medicine drugs.
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Research progress on animal models of malignant mesothelioma

YONG Sheng, JIN Dacheng, dong xin chun, GOU Yunjiu

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Malignant mesothelioma cases are rare and have a long incubation period, making it difficult to conduct clinical research.? Animal models of malignant mesothelioma are crucial for experimental research and elucidating the pathogenesis for malignant mesothelioma. Common animal models include spontaneous, inducible, transplantable, and genetically engineered models, but the applicability of different animal models varies. This article reviews studies related to the establishment of animal models of malignant mesothelioma in the past 10 years and summarizes the recent progress in the establishment of four animal models of malignant mesothelioma from three aspects: modeling methods, modeling results, and model advantages and disadvantages. This review summarizes and analyzes the current progress in the establishment of inducing animal models of malignant mesothelioma, this provides a reference for malignant mesothelioma basic research using animal models.
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The effect of ovariectomy on cognitive function and hippocampal estrogen receptor expression in APP/PS1 mice

congchao, Gu zuxi, Wu panqing, Chen siru, Lin guangyao, Xu lianwei

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Abstract: Objective: The present study aims to observe the changes in Aβ deposition and estrogen receptor levels in the hippocampal tissue of female APP/PS1 mice with Alzheimer's disease (AD) after ovariectomy (ovx) at different ages. Method: 3-month-old APP/PS1 female mice were divided into 3 groups, with 20 mice in each group. 10 mice in each group were treated with bilateral ovarian resection via abdominal surgery as the model group. 10 mice in the control group (sham group) were treated with only equivalent volume fat removed from the corresponding area during surgery as the control group (sham group). Feed APP/PS1 mice at different ages for 3 months, 4 months, and 5 months as the 6-month, 7-month, and 8-month age groups. Evaluate the cognitive function of APP/PS1 mice at different ages using Morris water maze. Observe the morphological changes of intracellular organelles in the CA1 area of the hippocampus of the two groups of mice using transmission electron microscopy. Immunofluorescence staining is used to detect the deposition of Aβ in the CA1 area of the hippocampus of the two groups of mice. ELISA is used to detect the levels and contents of serum estrogen, ROS, SOD, and MDA, and Western blot is used to determine the expression levels of ERα and ERβ. Result: As the age increased, the escape latency of the ovx group mice in the water maze test gradually prolonged, the deposition of Aβ in the hippocampal CA1 area increased, the mitochondrial swelling of hippocampal neurons increased, and a large amount of lipofuscin and amyloid deposition were observed in the cytoplasm. The serum estradiol level and SOD activity decrease, while the ROS level and MDA content increase. The expression of ERα and ERβ in hippocampal tissue decreases. Conclusion: Ovariectomy in mice with low estrogen status may exacerbate hippocampal Aβ deposition and age-dependent cognitive decline.
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The Effect of Western Dietary Feed on the Model of Atherosclerosis in APOE Mice

wanglei

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Objective To study the rapid modeling of Western dietary feed and its impact on biological indicators and histopathology in APOE-/- mice.Methods Using 48 female、48 male APOE-/- mice, 48 female、48 male C57BL/6J mice was tested and divided into 8 groups, namely the APOE-/- ordinary breeding feed group (24 female and 24 male each）、 APOE-/- Western Dietary Feed Group (24 female and 24 male each）、 C57BL/6J ordinary breeding feed group (24 female and 24 male each）、C57BL/6J Western Dietary Feed Group. Feed from 3w until the end of the experiment at 20w. After the experiment, serum was collected for biochemical indicators, and the aorta was separated for gross oil red staining and analysis. The root of the aorta was paraffin sectioned and HE stained. Results Western diet did not significantly increase APOE- / -body weight, western diet could significantly improve the blood lipid index of APOE- / -mice, total cholesterol, low density lipoprotein, high density lipoprotein, promote the formation of atherosclerotic plaque, male mice are suitable for the modeling of gross aortic plaque, and female mice are suitable for plaque modeling of aortic arch root. Conclusion Western dietary diet can promote APOE- / -mice, increase the aortic plaque area ratio, shorten modeling time, and improve modeling uniformity.
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Screening of antidiarrhea active parts and study on the “spectrum-effect” relationship of salt-processed Alpiniae oxyphyllae Fructus -Foeniculi Fructus medicines

li hongwei

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Objective To screen the antidiarrheal active components of salt-processed Alpiniae oxyphyllae Fructus -Foeniculi Fructus medicines (YZYH), and speculate on its pharmacological substance basis and quality markers through the study of the “spectrum-effect” relationship. Methods A model of diarrhea due to spleen and kidney yang deficiency was established in rats, and the pharmacodynamic indexes of YZYH on petroleum ether, ethyl acetate, and water fractions were measured, including behavioral indicators such as body weight, anal temperature, diarrhea incubation period, diarrhea index, and biochemical indicators such as NOS, cGMP, and CPK; Establish HPLC chromatograms of different parts of YZYH, and use grey correlation method to analyze the “spectrum-effect” relationship between the chemical components of each part of YZYH and the antidiarrheal efficacy indicators. Results In the aspect of pharmacodynamics, the ethyl acetate fraction had a better regulatory effect on the activities of DIP, DI, and serum NOS, cGMP, CPK in model rats compared to other treatment groups; In terms of the study of the “spectrum-effect” relationship, the correlation between components such as anisaldehyde and chromatographic peak 4 with the five efficacy indicators of DIP, DI, NOS, cGMP, and CPK was strong. Conclusion The ethyl acetate fraction was the active anti diarrhea fraction of salt-processed Alpiniae oxyphyllae Fructus -Foeniculi Fructus medicines. Its mechanism may be realized by regulating gastrointestinal peptide hormones such as MTL and GAS, inhibiting gastrointestinal hyperfunction, regulating energy metabolism and immunity, and cytoprotection; The ingredients such as anisaldehyde and peak 4 served as the material basis for its antidiarrheal effect, and anisaldehyde and peak 4 could be used as quality indicators for the medicines.
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Exploring the analgesic initiation mechanism of tuina on the dorsal root ganglion of minor CCI model rats via the TRPV1/TRPA1 -cGMP signaling pathway

YANG Zhenjie, YU Tianyuan

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Objective: To explore the analgesic initiation mechanism of the three-manipulation and three-acupoint of tuina on rats with minor CCI model. Methods: 56 SD rats were randomly divided into 8 groups: normal group, sham group, model 1 group, model 2 group, tuina 1 group, tuina 2 group, tuina 1 + TRPA1 antagonist group, and tuina 2 + TRPV1 antagonist group. The model group, tuina group, and tuina + antagonist group were established with minor CCI models. The tuina group and tuina + antagonist group received the three-method three-point intervention (point method, dial method, kneading method, Yinmen point, Chengshan point, Yanglingquan point) 7 days after modeling. The model group and sham group were subjected to grasping restraint, and the normal group was not intervened. After the intervention, each group was tested for changes in thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) to detect different types of pain. The nitrate reductase method was used to observe the NO content in the DRG of each group on the modeling side, and ELISA, Western blot, qPCR, and other methods were used to observe the changes in the protein and gene expression of the TRPV1/TRPA1-NO-cGMP-PKG signaling pathway in the DRG of each group on the modeling side. Results: Compared with the model group, behavioral testing showed that the MWT and TWL were prolonged to varying degrees in the tuina 1 group and tuina 2 group. The expression of TRPV1, TRPA1, NO, sGCβ, cGMP, and PKG1 in the DRG of the tuina 1 group, tuina 2 group, tuina 1 + TRPA1 antagonist group, and tuina 2 + TRPV1 antagonist group were significantly decreased. Conclusion: Tuina intervention can effectively improve the symptoms of thermal and mechanical hyperalgesia caused by peripheral nerve injury after one intervention. Tuina can exert immediate analgesic effects through the TRPV1/ TRPA1-NO-cGMP-PKG signaling pathway.
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Establishment of intestinal polyps animal model with Apc-KRAS-Cre genetic mutation

TAN WEI SHAN, WANG SHU YUN, WANG HAO YUE, SUN KE XIANG, GAO JIA MIN, DENG WAN LI

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Abstract: Objective To create the mice model of colorectal polyps with Apc-KRAS-Cre gene mutation with the best tamoxifen induction method. Methods C57CL/6J mice were used as healthy control group. Genetically mutated mice were grouped and intraperitoneally injected with tamoxifen of different concentration and dosage for different durations. The survival rate and change in weight of mice were observed. 4 weeks post-tamoxifen induction, mice were euthanized and the length of colons, amount and size of intestinal polyps were observed. Through H&E staining, the histopathological changes of the intestinal tissue and polyps were observed. Results The survival rate of male mice is higher than that of female mice(P<0.05), while the survival rate between the 4 groups shows statistical significance(P<0.05). When compared to the healthy control group, the changed in weight and the length of the colons of each group of mice showed statistical significance（P<0.001）, (P<0.05), the difference in weight change between group 1 and group 2, group 2 and group 3, group 1 and group 4; the difference in length of colons between group 1 and group 3 showed statistical significance (P<0.05). Each group of mice induced by tamoxifen injection showed intestinal polyps of varying sizes, mostly occuring on the distal end of the colon, while those in group 3 and group 4 have a higher amount and larger polyps. Comparing to healthy control group, histopathology of tamoxifen-induced mice showed growth of intestinal polyps, with uneven and misaligned glandular and epithelial arrangements, loosely-packed intestinal mucosal barrier and irregularly-distributed crypts, group 3 and group 4 mice showed signs of inflammation, while regions of group 4 showed necrosis of cells. Conclusion Tamoxifen-induced Apc-KRAS-Cre mice models have been successfully established, with group 3 induction method being the most suitable.
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The method and improvement of implantable osmotic pump for intraventricular drug administration in rats

SUN Heyong, ZHANG Gangli, WU Jiliang, CAO Peili, LI Shuo, SUN Haoqin

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Objective To introduce and enhance the experimental technique for intraventricular drug delivery via an implantable osmotic pump. Methods Eight-week-old male SD rats were selected, and the requisite equipment and reagents were prepared. Initially, the osmotic pump was assembled and brought to operational status before conducting the implantation surgery. Following anesthesia, the animals underwent skin preparation, and the skull's upper surface was surgically exposed. Utilizing a brain stereotaxic apparatus, a point directly above the ventricle was located, and a small hole was drilled at that location with a high-speed cranial drill. Subsequently, the pump body was implanted subcutaneously in the neck, the needle was inserted into the drilled hole, and secured with dental cement. Once solidified, the needle base was removed, the subcutaneous soft tissue and scalp were sutured in layers, and the animal was returned to its cage for isolated rearing. Results The osmotic pump was successfully implanted subcutaneously in the rat's neck, the needle was securely affixed to the skull, and the catheter interface remained intact. Upon examination of the extracted brain tissue, no significant hematoma was observed around the puncture site or needle tract, and the presence of blue dye in the ventricular and adjacent tissues indicated successful drug delivery to the ventricle. Conclusion The introduction of a brain stereotaxic apparatus for localization assistance, coupled with enhancements to the operational procedure, has rendered the implantation process more accurate and safe, achieving a high success rate in intraventricular drug administration.
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The positive effect of the good operation of the CNAS-CL06 system on the scientific nature of animal experimental

maliying, wanghong, guomeng, liangchunnan, yuebingfei, gongwei

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The animal laboratory of pharmaceutical and biological product inspection and testing institutions undertakes important basic support tasks for testing and scientific research work. This article summarizes the management experience accumulated in the operation of our institution's CNAS-CL06 quality management system, providing reference for similar institutions in operation to ensure the scientific nature of animal experimental data. In order to ensure the scientific nature of animal experimental data, the experimental animal institution has successfully passed the CNAS accreditation of the experimental animal institution, and has completed rectification on time during on-site supervision and evaluation. At the same time, regular self inspections of the system have been carried out in accordance with the "Quality and Capability Accreditation Guidelines for Experimental Animal Breeding and Use Institutions" (CNAS-CL06). During the self inspection process, we examined issues while improving the content of the system. Starting from animal procurement, occupational health and safety, animal disease treatment and care, and facility operation emergency drills, we enriched the content of the quality management system, ensured the continuous and effective operation of the system, and ensured the effectiveness and standardization of animal experiment data. At the same time, we contributed to the management ideas of our institution in sharing animal experiment platforms, Provide hardware and software support for the construction of cloud platforms for animal experiments. This article aims to explore and form a quality management model for the experimental animal industry based on practical work, Being focused in the standardization strategy， continue to deepen the reform of standardization work, and give full play to the basic and strategic role of standardization in the modernization of the animal experiments system.
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Management and experience exchange of three-level laboratory of animal biosafety in University

tianxiaoyan, Yangwenhao

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Animal Biosafety Level 3 laboratory (Absl-3) is the basic supporting platform of the national biosafety system for new and recurrent infectious diseases, it provides an important hardware basis for the study of biological characteristics, prevention and treatment of highly pathogenic microorganisms. ABSL -3 laboratory also provides an important support for the combination of production, teaching and research. The normal operation of ABSL -3 laboratory and the smooth development of scientific research work can not be separated from the careful planning and effective implementation of emergency planning work. In addition, the reasonable design and layout of the laboratory is also a key factor to avoid and reduce accidents. Based on the practice of construction and management of animal biosafety level III Laboratory in Yangzhou University, starting with the preparation of emergency plan, this paper introduces the points for attention in the layout of laboratory construction, daily operation and emergency treatment, it is expected to provide useful reference for biosafety laboratories operating various pathogenic microorganisms in the future.
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Research progress on the mechanism of miR-138-5p in osteoarthritis

Wu Liangbin, Weng Jian, Li Aikang, Qi Tiantian, Zhang Geng, Zeng Hui, Yu Fei

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MiR-138-5p, as a microRNA, plays an important regulatory role in the pathogenesis of osteoarthritis. It regulates various biological processes such as inflammation, cell apoptosis and proliferation, and matrix degradation in osteoarthritis by modulating signaling pathways including NF-κB, Wnt/β-catenin, and PI3K/AKT. This review summarizes the research progress on the mechanism of miR-138-5p on osteoarthritis.
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Preliminary Discussion on the Existing Issues of Ethical Review of Laboratory Animals Welfare in Medical Research

wang xiao xiao

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Laboratory animals have played an important role in maintaining human health. Advocating ethics and welfare of laboratory animals is a dual manifestation of technological progress and human civilization, and establishing ethical review of laboratory animals welfare is an effective guarantee for achieving the welfare of laboratory animals. Although the construction of animal welfare ethics in China has developed rapidly in recent years, there are still many problems that need to be continuously improved at this stage. This article is based on the understanding and recognition of the ethical concepts related to the welfare of laboratory animals. It summarizes and discusses some issues discovered during the ethical review of laboratory animals welfare in medical research, and puts forward personal suggestions for exchange and mutual refrence.
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Research progress of the role of ferroptosis in infectious diseases

YUE Linzhi, MA Tao, DAI Yumei, DU Wenya, WANG Guofu, WU Lixian

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Ferroptosis is a newly discovered mode of programmed cell death characterized by the accumulation of intracellular iron-dependent lipid peroxidation. At present, the research on ferroptosis mainly focuses on the field of cancer, but more and more evidence shows that ferroptosis is related to the occurrence of infectious diseases, and ferroptosis has been found in Corona Virus Disease 2019, tuberculosis and cryptococcal meningitis and other diseases. This article reviews the role of ferroptosis in infectious diseases, in order to provide new ideas for the prevention and treatment of ferroptosis related infectious diseases.
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Oxidative stress-based exploration of the zebrafish in cardiovascular disease and research progress

Fei Tiantian, Liang Tengyun, Wu Hong

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Cardiovascular disease has been a serious threat to human life, and oxidative stress is an important factor in the occurrence and development of cardiovascular disease. The construction of reliable animal models of oxidative stress is helpful for in-depth study of the pathogenesis of cardiovascular diseases and the development of therapeutic drugs. Zebrafish, with the advantages of easy reproduction, short developmental cycle, transparent embryos for easy observation, and highly homologous cardiovascular gene background with humans, has often been used in recent years for research related to the field of cardiovascular diseases. In this paper, we focus on the application of zebrafish oxidative stress model in cardiovascular disease and the related research progress, which provides reference for its further application in cardiovascular disease related research.
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Effects of ezrin protein on Helicobacter pylori-induced nodular gastritis

Wang Peng, Zhu Hong-wei, Jiang Shuy-uan, Liu Xiao-lei, Gao Bin, Shao Guo

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The ERM protein family (ezrin, radixin, and moesin) plays a pivotal role in cell morphology, migration, and signal transduction. Ezrin, a prominent member of this family, is highly involved in these processes. Particularly, the phosphorylation of ezrin is crucial, as it regulates the interaction between ezrin and the actin cytoskeleton. This interaction is a key mediator of cytotoxicity in host cells infected with Helicobacter pylori, significantly impacting cell morphology. This review comprehensively summarizes the multifaceted role of ezrin protein in Helicobacter pylori-induced nodular gastritis. It delves into the relationship between ezrin"s structure, function, phosphorylation, and signaling pathways in the context of nodular gastritis. Moreover, this paper highlights ezrin protein as a potential therapeutic target, offering novel insights for the prevention and treatment of nodular gastritis.
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Research progress on mitochondrial-associated membranes (MAMs) in the pathogenesis of Alzheimer's disease

liushiyu, xuyumin, xuhongcai, lufangmei

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Mitochondrial-associated membranes (MAMs) is a special domain for communication and material exchange between mitochondrial outer membrane and endoplasmic reticulum membrane, which performs various biological processes of cell life activities under different conditions. MAMs dysfunction-mediated calcium homeostasis imbalance, endoplasmic reticulum stress, mitophagy defects, mitochondrial fission/fusion dynamics imbalance, lipid metabolism disorders, and inflammatory responses are the key pathogenesis of Alzheimer’s disease (AD). This article reviews the structure and function of MAMs, their involvement in AD pathology and drug intervention targets, and discusses the role of MAMs in the pathogenesis of AD and the latest mechanism research progress, in order to provide new ideas for the prevention and treatment of AD.
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Research progress on Mitochondrial Damage in Inflammatory Bowel Disease

WU Xiaoting, XUAN Ang, SHEN Juan

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Inflammatory bowel disease (IBD) is a chronic and relapsing inflammatory disorder of the gastrointestinal tract that poses a significant threat to human health. The pathogenesis of IBD remains unclear and is believed to involve various factors such as genetics, immune dysregulation, and environmental triggers. In recent years, increasing evidence has highlighted the role of mitochondrial damage and dysfunction in the development of IBD. This review provides a comprehensive review and overview of studies related to mitochondrial damage in inflammatory bowel disease, focusing on the effects of mitochondrial oxidative stress damage, autophagy dysfunction, kinetic disturbances, and respiratory defects on IBD. The aim is to identify potential therapeutic targets and provide new insights for the scientific prevention and treatment of IBD.
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miR-22-3P targets GSDMD to inhibit homocysteine induced pyroptosis of vascular smooth muscle cells

ZHONG Yingyi, Ding Ning, Wang Yicheng, Liu Chao, Ma Shengchao, Xiong Jiantuan, Hao Yinju, Bai Zhigang, Jiang Yideng

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Objective To investigate the effect of miR-22-3p on pyroptosis of vascular smooth muscle cells induced by homocysteine. Method Human vascular smooth muscle cells (VSMC) were cultured in vitro and divided into the Control group (0 μmol/L Hcy) and the Hcy group (100 μmol/L Hcy). After intervention, Western Blot was performed to detect the expression of Pro Caspase-1, GSDMD, N-GSDMD and NLRP3. The qRT-PCR was used to measure the expression of miR-22-3p. ELISA was utilized to survey the concentration of IL-1β and IL-18 in the supernatant. After transfection with control of miR-22-3p (miR-22-3p-NC) ，mimic of miR-22-3p (miR-22-3p-mimic) , and inhibition of miR-22-3p (miR-22-3p-inhibitor) ， to observe the change of VSMC pyroptosis induced by homocysteine. Result Compared with Control group, the expression levels of Pro Caspase-1, GSDMD, N-GSDMD, NLRP3 in VSMC of Hcy Group were increased (P<0.05) , the concentration of Il-1β,IL-18 were higher (P<0.01) , and the relative expression level of miR-22-3p was lower (P<0.01) . After transfection with miR-22-3p-mimic, the expression levels of Pro Caspase-1, GSDMD, N-GSDMD and NLRP3 in VSMC decreased significantly (P<0.01) , and the concentrations of il-1β and IL-18 decreased significantly (P<0.01) . After transfection with miR-22-3p-inhibitor, the expression levels of Pro Caspase-1, GSDMD, N-GSDMD and NLRP3 in VSMC increased significantly (P<0.01) , and the concentrations of il-1β and IL-18 were higher (P<0.05) . Conclusion miR-22-3p delayed Hcy induced VSMC pyroptosis.
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Dioscin promotes apoptosis of HepG2 cell through inhibiting Wnt/β-catenin signaling pathway

liangyuqiong, huangqing

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Objective ?To detect the effects of apoptosis on HepG2 cell induce by Dioscin and its possible mechanism of anti-hepatocellular carcinoma. Methods The HepG2 human hepatocellular carcinoma cells were exposed with 0.25,0.5,1,2,4,6,8 μmol/L dioscin, and then the cell proliferation ability was measured with
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Evaluation of pulmonary arteriolar lesions in rats induced by cigarette smoke exposure combined with Klebsiella infection

REN Zhouxin, ZHAO Peng, LI Jiansheng

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Objective To analyze the morphological and structural changes of pulmonary arterioles in rats induced by smoke exposure combined with Klebsiella infection, and to evaluate the severity of pulmonary arteriolar lesions. Methods Pulmonary arteriolar images of lung sections of control rats and model rats induced by smoke exposure combined with Klebsiella infection were analyzed by qualitative and quantitative methods. Victorian blue stained sections were used for the detection of pulmonary arteriolar muscularization, vascular wall thickness, vascular occlusion score, intima thickness and media thickness of muscular arterioles, and neointima proliferation. HE stained sections were used for the observation and detection of inflammatory cell infiltration and plexiform lesions around arterioles. VG stained sections were used for the observation of collagen fibers in the intima and detection of the percentage of collagen fibers area in arteriolar wall area. Based on the above results, the degree of pulmonary arteriolar lesions was rated according to the Heath-Edwards criteria. Results For diameter ≤ 50 μm arterioles, compared with the control group, the percentage of non-muscular vessels was significantly decreased, the percentage of muscular vessels was increased, the percentage of partial muscular vessels was not significantly different, the wall thickness of non-muscular vessels and the wall thickness of muscular vessels were significantly increased, and the occlusion scores of both non-muscular and muscular pulmonary arterioles were significantly increased in the model group. For 50 < diameter ≤ 100 μm arterioles, compared with the control group, the percentage of non-muscular vessels was significantly decreased, the percentage of muscular vessels and the percentage of partial muscular vessels were not significantly different, the wall thickness and occlusion score of muscular vessels were significantly increased, the wall thickness and occlusion score of non-muscular vessels were not significantly different in the model group. Compared with the control group, the intimal thickness and media thickness were significantly increased and the perivascular inflammatory infiltration score was significantly increased in muscular arterioles with both diameter of ≤ 50 μm and 50 < diameter ≤ 100 μm of the model group. In the control group (n=9), only one section with neointimal lesion was found with 1.61 % of proliferation degree of neointima. In the model group (n=10), five sections had neointima lesion from 1.04 % to 17.14 % of proliferation degree of neointima, respectively. No plexiform lesions were found in a11 sections. For pulmonary arterioles with diameter of ≤ 100 μm, there was no change in the expression of intimal collagen fibers in the model group compared with the control group, and there was no significant difference in the percentage of collagen fiber area in the vessel wall area. According to the Heath-Edwards criteria, the pulmonary arteriole lesions in the model rats did not reach grade III. Conclusions The model rats show pathological manifestations such as pulmonary arteriolar muscularization, thicking intima and media, and mild to moderate inflammatory reactions around arterioles. The low degree of neointimal proliferation and collagen fiber in the vascular wall and no plexiform lesions suggest that the model is up to grade II lesion according to the Heath-Edwards criteria.
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Progress of animal experimental research on the treatment of Henoch-Schonlein Purpura with TCM

Su Hang, Zhang Shuzi, REN Xianqing

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Henoch-Schonlein purpura (HSP) is the most common systemic vasculitis in childhood, which is called "spot toxin", "purpura wind" and "grape plague" in TCM. Modern medical research suggests that the disease is related to factors such as imbalance of cellular and humoral immunity, abnormal secretion of cytokines, disturbance of coagulation and fibrinolysis mechanism, but the exact pathogenesis is still unclear. The incidence of this disease increasing year by year, with a high recurrence rate and a high proportion of kidney damage, which seriously affects the physical and mental health of the[[基金项目] 国家自然科学基金（82374519,U2004107）；河南省青年科学基金项目（232300421309）；河南省中医药科学研究专项（20-21ZY2161） [作者简介] 苏杭（1988-），男，副主任医师，硕士研究生，研究方向：中医药防治小儿肾病及风湿免疫性疾病 Email：362504467@qq.com [通信作者] 任献青（1973-），男，教授，主任医师，博士研究生，研究方向：中医药防治小儿肾病及风湿免疫性疾病 Email：renxq723@163.com ] affected children and poses a great social risk. Numerous studies have shown the clinical efficacy of TCM in treating HSP, but its mechanism of action is not yet completely clear. In recent years, a large number of animal experiments have been carried out to study the efficacy mechanism of TCM with the establishment of animal models of HSP, but there is a lack of a more systematic and detailed reviews. Therefore, the animal experimental literature related to the treatment of HSP with TCM in the past decade were reviewed, and the effects of TCM in alleviating abnormal glycosylation of IgA1, regulating the immune imbalance of Th1/Th2 and Treg/Th17 cell, reducing circulating immune complex, and inhibiting inflammatory responses were compiled and summarized, In order to providing a reference for the further research on TCM for the treatment of HSP and inspiring new research ideas.
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Progress of miRNA in the pathogenesis of pulmonary hypertension

LI Yanjin, XIAO Liying, WU Daoxiong, GUAN Rong, YAN Chunlang, LEI Wen

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Pulmonary hypertension(PH) is a progressive disease characterized by pulmonary vascular remodeling. Current treatments for PH remain suboptimal, and there is an urgent need to better decipher the underlying pathomechanisms in order to find new therapeutic targets. miRNA(microRNA) are key components in the post-transcriptional machinery that mediate cellular functions mainly by regulating the expression of downstream target genes. Numerous in vivo and in vitro studies have demonstrated the involvement of miRNA and their regulators in PH development. However, there is no unified conclusion about the mechanism of miRNA regulation of pulmonary vascular remodeling. Therefore, this article provides a review on the mechanisms of miRNA in PH in recent years.
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[ ]Establishment of sterile golden hamster model

GUO ya-xi, duxiaopeng, liukaihui, wangzhaohua, zhuhua

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Abstract：【Objective】 A sterile golden hamster model was established by cesarean section purification method.【Methods】The donor female golden hamsterrats selected SPF grade golden hamster, male and female were mated 1:1, and were separated from male rats after meeting the peg. The cage time of the surrogate mothers rats should be one week earlier than that of the donor mothers. The pregnant parturient golden hamsters underwent hysterectomy in a super-clean workbench. at the time of delivery, andThe uterus were transferred into isolation kits for purification. The uterus wasand stripped to obtain the milk mice to be purifiedfor purification, . and sterile Sterile ICR mice and sterile SD rats were respectively used for milk replacement, and transferred to isolation kits for feeding after successful milk separation. The sterility status of feeding isolation kits is tested monthly. 【Results】Three caesarean sections were performed, but the first and second lactation failed. The third milk replacement was successful, and 18 young rats hamsters were obtained with survival rate of 88% and survival rate of 66% after milk release. All of them hamsters were qualified according totested by GB /T 14926.41-2001.【Conclusion】By cesarean section purification technique, sterile ICR mice and sterile SD rats were used for milk replacement to achieve the purpose of microbial purification, and the sterile golden hamster model was obtained.
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Inhibitory effect of 17-DMAG on PD-1 humanized mouse liver cancer transplantation tumor

LI Xiaojuan#, XIU Ye#, LI Xingjie, SUN Yanfeng, LI Ruisheng

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Objective Explore the inhibitory effect of 17-DMAG on the growth and angiogenesis of PD-1 humanized mouse liver cancer transplantation tumor. Methods 30 PD-1 humanized mice were selected and human HepG2 cell suspension was injected into the subcutaneous tissue of the right inguinal region of the mice to construct a human liver cancer transplant tumor model. Tumor bearing humanized mice were randomly divided into three groups (10 mice per group): ① model group (injected with 10 mg/kg of physiological saline); ② 17-DMAG group (intraperitoneal injection of 17-DMAG at 25 mg/kg, 3 times/week); ③ The cisplatin group (intraperitoneal injection of 20 mg/kg, 2 times per week), the experiment lasted for 4 week. After injection, the length and short diameter of the humanized mouse transplanted tumor were measured to calculate the volume, and the tumor mass were measured to calculate to calculate the tumor inhibition rate. At the same time, immunohistochemical methods were used to detect the expression of CD31 (tumor microvessel density, MVD) and vascular endothelial growth factor (VEGF) in tumor tissue. Results The tumor volume and mass of the 17-DMAG group and the cisplatin group were significantly reduced compared to the model group (P<0.05), and the tumor inhibition rate of the 17-DMAG group was slightly higher than that of the cisplatin group. However, there was no significant difference in tumor mass, volume, and tumor inhibition rate between the 17-DMAG group and the cisplatin group. The number of MVD labeled microvessels and VEGF expression in the 17-DMAG group and cisplatin group were lower than those in the model group (P<0.05), and the 17-DMAG group was also lower than those in the cisplatin group (P<0.05). Conclusions 17-DMAG can inhibit the growth of humanized mouse liver cancer xenografts by reducing the expression of vascular endothelial growth factor in liver cancer xenograft tissue, thereby inhibiting the generation of tumor neovascularization.
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Research progress of liver fibrosis based on Notch signal pathway

jishaoxiu, zhanghengyao, zhangyuan, wenli

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Liver fibrosis is a necessary pathological process from chronic liver disease to cirrhosis and even liver cancer. The occurrence of liver fibrosis is a highly integrated and dynamic pathological process resulting from the interaction of many cells and cytokines. Notch signaling pathway is an evolutionarily conserved intercellular signal transduction mechanism, which plays an important role in regulating the development and tissue renewal of multicellular animals. More and more studies have shown that Notch signal transduction participates in the formation of liver fibrosis through a variety of ways. Therefore, this paper reviews the role of Notch signaling on cells involved in the formation of liver fibrosis to explain the function of Notch signaling pathway in liver fibrosis.
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Construction and evaluation of an immunosuppression-mediated model of invasive Aspergillus niger lung disease in rats

TANG Zining, CHEN Xiangchi, LIU Xuewu, ZHOU Zhimin, LI Qiao, XIAO Sa, JIANG Dejian, PENG Dongdong

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Abstract: Objective This study establishes a model of invasive Aspergillus niger lung disease in immunosuppressed rats to provide theoretical support for the pharmacodynamic evaluation of anti-invasive pulmonary aspergillosis drugs and mechanism studies. Methods Sixty SD rats were randomly divided into a normal control group, a cyclophosphamide control group, and a cyclophosphamide + fungal infection low, medium, and high dose group, with 12 animals in each group. General clinical observations were performed daily, and the serum levels of immunoglobulin G (IgG), immunoglobulin M (IgM) and serum galactomannan (GM) were detected by ELISA on the 3rd and 7th days of modeling, respectively, as well as the ratio of CD4+ and CD8+ cells and the content of white blood cells (WBCs) and neutrophils (Neu) in peripheral blood, and the load of Aspergillus niger in alveolar lavage were observed simultaneously and morphological changes of rat lung tissue. Results The rats in the cyclophosphamide control group and the cyclophosphamide+fungal infection group showed reduced voluntary activity and erect hair after modeling, and the rats in the cyclophosphamide+fungal infection group were accompanied by shortness of breath and audible wet rhonchi in the lungs; compared with the normal control group, the rats in the cyclophosphamide control group showed significant reductions in the levels of CD4+, WBC, Neu, IgG and IgM in the blood, and the proportion of CD8+ was significantly higher (P<0.05, P<0.01); compared with the cyclophosphamide control group, rats in the cyclophosphamide+fungal infection group showed significant decreases in blood levels of CD4+, WBC, Neu, IgG, IgM, and significant increases in levels of CD8+, GM, and the alveolar lavage load of Aspergillus niger (P<0.05, P<0.01); and the lung tissues of the cyclophosphamide+fungal infection rats in the low-, medium-, and high-dose groups showed mycelial distribution and destruction of alveolar epithelium, increase of bronchial epithelial cup cells in the alveoli, and infiltration of inflammatory cells, and the degree of lesions was positively correlated with the modeling dose. Conclusion In this study, we used Aspergillus niger combined with cyclophosphamide immunosuppressant to construct a model of invasive Aspergillus niger lung disease, and the duration of the disease was positively correlated with the concentration of the bacterial fluid and the modeling time, which confirms that the cellular immunity plays an important role in the pathogenesis of the disease, and at the same time the immunoglobulin can also affect the development of the disease process of invasive pulmonary aspergillosis, and it is speculated that the pathogenesis of invasive pulmonary aspergillosis may be related to the level of immunoglobulin in the humoral immunity.
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Educational reform of animal experiment course for medical graduate students: cultivating students" ability of active learning and innovative thinking

MIAO Jinxin, MIAO Mingsan

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Animal experiment course is an important practice teaching link in medical and life science. It is of great significance to the cultivation of students' experimental skills, scientific thinking and innovative ability. However, the traditional teaching method of animal experiment course has the problems of single content and low participation of students. In order to promote students' active learning and innovative thinking, this paper aims to discuss the teaching reform of animal experiment course. Firstly, the paper introduces the background and significance of the teaching reform of animal experiment course and emphasizes its importance to the cultivation of students' ability. Secondly, it puts forward the principles of teaching reform of animal experiment course, including student-centered, paying attention to problem solving and practice exploration, and promoting interdisciplinary integration. Then, from the aspects of course design, teaching methods and evaluation methods, this paper expounds the concrete measures of teaching reform of animal experiment course in detail. In the course design, attention should be paid to the selection of challenging and exploratory experimental projects, and students' interests and professional needs should be fully considered. On teaching methods, students should be encouraged to actively participate in, exploration and cooperation, and guide them to problem solving and the cultivation of innovative thinking. In terms of evaluation methods, diversified evaluation methods, such as experimental report, group discussion and project presentation, should be used to comprehensively evaluate students' comprehensive ability and innovative thinking. Finally, through practical verification and effect evaluation, the experience and results of the teaching reform of animal experiment course were summarized, and suggestions for further improvement were put forward.
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Progress of mitochondrial mass control system in the pathogenesis of septic cardiomyopathy

xie you cheng, xiao shu fang, lin xue mei, chen shun, yu xiaohui

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Septic cardiomyopathy (SIC) is a frequently observed organ dysfunction in sepsis, characterized by high mortality and poor prognosis. Understanding the complex pathogenesis of SIC and developing effective therapeutic tools are critical issues that require attention. Previous studies have demonstrated the significant role of mitochondrial dysfunction in the development of SIC. In the presence of SIC and resulting mitochondrial dysfunction, aberrant regulation of the mitochondrial quality control system (MQC) can exacerbate cardiomyocyte injury. Recent studies have demonstrated that the mitochondrial quality control system (MQC) maintains mitochondrial dynamic homeostasis through its regulation of mitochondrial biogenesis, fusion/fission, and autophagy. Consequently, this article provides an overview of the role of MQC in SIC pathogenesis, reviews the latest studies, and analyzes its potential as a therapeutic target.
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Animal Model Analysis of Atherosclerosis Based on Clinical Symptoms in Traditional Chinese Medicine and Western Medicine

Luo Shiwen, Yang Dongmei

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Objectives Based on the clinical characteristics of atherosclerosis in? traditional Chinese medicine (TCM) and Western medicine, this paper analyzes the common animal models of atherosclerosis(AS), and the coincidence of the models with clinical characteristics was scored, in the hope of providing new ideas and reference for studying animal models of AS. Methods This paper reviewed the varieties, modeling methods, modeling principles and characteristics of common animal models of AS. Moreover, according to the clinical diagnostic criteria and symptom characteristics, the similarity of the common animal models was analyzed. Results High-fat feeding type, mechanical injury combined with high-fat feeding type, genetic engineering combined with high-fat feeding type, chemical induction combined with high-fat feeding type and the models of AS combining Chinese clinical syndrome and western disease are widely established in animal models of AS. Comparative analysis result showed that balloon injury combined with high fat feeding type, ApoE receptor knockout mouse combined with high-fat diet type and phlegm and blood stasis type in models of disease and symptom combination showed a comparatively high level of clinical agreement between Chinese and western medicine. Conclusions Presently, most of the animal models of AS had the high degree of coincidence with western medicine, evaluation criterion is dominated by the western medicine as well, which lack of the combination between disease and syndrome, so as to hinder development of concept of Wholism and treatment by differentiation of syndromes in TCM. Therefore, establishing the exact and high coincidence degree animal model, which combines the disease and its TCM symptoms, is the top priority of studying the prevention and treatment of atherosclerosis.
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The role of UBC9 mediated SUMO modification in homocysteine-induced pyrodeath of macrophages

Ma Lingju, Chi Hongyang, Wu xinxue, Ma Fujun, Tian Yancheng, Zhao Caiqi, He Tianyu, Peng hongjian, JiangYideng, Yang Li, Huang Hui, Ma Shengchao

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Objective To study the role of ubiquitin Conjugating Enzyme 9 (UBC9) in the pyrodeath of homocysteine-induced macrophages mediated by SUMO modification. Methods Firstly, The effects of homocysteine at different concentrations (0μmol/L, 50μmol/L, 100μmol/L, 150μmol/L, 200μmol/L) on the viability and pyrodeath of mouse macrophages (RAW264.7) were detected by CCK-8 and Western blot ;Western blot was used to detect the expression levels of UBC9, SUMO-modified protein SUMO-1 and inflammatory cytokine IL-1β in different groups of cells. qRT-PCR was used to detect the mRNA expression of UBC9 before and after interference, and to detect the expression of UBC9, pyrogen-related protein and SUCO-1 after interference.Results After stimulation with 100μmol/L Hcy, the activity of macrophages was not affected, and the expressions of NLRP3 and caspase-1 were the most obvious (P < 0.05).Compared with Control group, the expression levels of and IL-1β were increased in Hcy group (P < 0.01), and the expression of SUMO-1 was increased (P < 0.01). Compared with Control group, UBC9 protein level and mRNA level in Hcy group were increased (P < 0.05). After transfection with si-UBC9, the expressions of NLRP3, caspase-1, IL-1β, UBC9 and SUMO-1 in si-UBC9+Hcy group were decreased compared with si-NC+Hcy group (P < 0.01). Conclusion Hyperhomocysteamine (Hcy) induces pyrodeath in macrophages, and its mechanism is related to the up-regulation of ubiquitin binding enzyme 9 to SUMO modification.
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Visualization Analysis of Research Progress on Neutrophils in Diabetes Based on Citespace

WANG Jin-xi, YU Rong, HUANG Juan, Liu Yangyang, ZHANG Tian-tian, LI Wei, YANG Hui, LEI Shi-hui

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Objective This article aims to clarify the development of neutrophils and diabetes fields, as well as the evolving characteristics, potential trends, and research hotspots of neutrophils in the diabetes field. Methods 2998 relevant literatures on neutrophils in the diabetes field indexed in Web of Science from 2010 to 2023 were retrieved, and visual analysis of the relevant literatures on neutrophils in the diabetes field was conducted using Citespace6.1.R6. Results Since 2012, publications on this topic have grown rapidly. Bayat Mohammad, Liu Tong, Amini Abdollah, Zhang Rui are high-yield authors, with 7 related articles published. China and Shanghai Jiao Tong Univ are the countries and institutions with the most published papers. The most influential journal in this field is "PLOS ONE", cited 1380 times. Literature co-citation analysis shows that the highest focus in topics related to diabetes currently are "extracellular trap" and "covid-19 patient". Co-occurrence analysis, clustering analysis, and keyword burst analysis indicate that "lymphocyte ratio" (13.08) and "neutrophil extracellular trap" (7.2) are the most researched topics in the field of neutrophils and diabetes. Literature in this field mainly focuses on "myocardial infarction", "endothelial", "oxidative stress", and "apoptosis". Conclusion This article analyzes the evolving trends of neutrophils in the diabetes field using Citespace, providing new insights for researchers to conduct future research in the diabetes field.
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Observation on the progress of the murine models of chronic ulcerative colitis induced by dextran sodium sulfate

KONG Weijiao, YAN Yiyue, ZHAO Peikai, MAO Xiaojian, WANG Ting

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Objective To investigate the progress of chronic Ulcerative Colitis (UC) model induced by dextran sulfate (DSS) in mice. Methods 2.5% DSS solution were provided ad libitum for 5 days, and tap water for were supplied for another 5 days in one induction cycle. Male C57BL/6 mice were undergone continuous three cycles of induction to establish the chronic UC model. The weight, disease activity index (DAI), organ index, colon length of mice and pathological changes of colon tissue were determined. The level of myeloperoxidase (MPO) in colon tissue, cytokines such as IL-1β in serum and colon were detected. Results During the three cycles, all mice had diarrhea, bloody stool, weight loss, colon length shortening and pathological injury, which were aggravated after DSS administration and relieved after drinking water administration. The content of MPO in colon gradually increased. The spleen index and the levels of IL-1β, IL-6, IL-17A, TGF-β and TNF-α in serum increased continuously or fluctuating. The contents of IL-1β, IL-6 and IL-17A in colon increased significantly in the first or second cycle, and then decreased slowly. There was no significant change in TNF-α in colon. The concentrations of IL-10 in serum and colon fluctuated up and down, and decreased to a level lower than or remained the same as that in normal group at the end of the experiment. Conclusion Chronic UC model can be successfully induced by 2.5%DSS solution, which is characterized by alternating symptoms of "attack-remission". The persistent systemic inflammatory reaction and overactivation of colonic neutrophils reflect the characteristics of recurrent UC to some extent. It can be used as a tool for analying the pathological changes and interrention opportunities of UC, as well as for discovering and evaluating UC therapeutic drugs.
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Research progress of ferroptosis in sepsis-associated acute lung injury

SHI Yanglin, YANG Jianya, CHANG Qingqing, WANG Qianqian, WANG Minghang, LI Suyun

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Sepsis is defined as life-threatening organ dysfunction caused by a dysregulated host response to infection, with an extremely high mortality rate. It is the main risk factor for acute lung injury (ALI). However, the pathophysiology and pathogenesis of sepsis-associated ALI are not fully understood, and effective drugs are extremely limited. Therefore, the urgent task is to explore the pathogenesis of sepsis associated ALI and attempt to discover effective intervention measures to improve the prognosis of sepsis associated ALI patients. In recent years, ferroptosis has been considered closely related to the pathological and physiological processes of sepsis associated ALI, and inhibiting related cell ferroptosis can effectively slow down the occurrence and development of the disease. In this paper, the therapeutic strategies targeting ferroptosis of related cells were reviewed, in order to provide reference for the research on ferroptosis of sepsis associated ALI and provide a new perspective for the treatment of sepsis associated ALI.
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Effect and mechanism of ICAM5 on alcohol dependence behavior of mice

Hu Jiajia, Yang Zhuanfang, Sun Xizhe, Yuan Juanjuan, Cheng Yan, Zhang Yu, Yin LITIAN

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【Abstract】 Objective We investigated the effects of ICAM5 in hippocampus of mice alcohol drinking preference, and potential mechanisms. Methods A alcohol two-bottle choice model was developed by 8-week-old male C57BL/6J mice, randomly divided to two groups, the control group and alcohol group. The protein expression of ICAM5 in the hippocampus, amygdala, and medial prefrontal cortex was detected. ICAM5 overexpressed adeno-associated virus was constructed and injected into the hippocampus by stereotaxic method. The expression level of ICAM5 protein in hippocampus was detected by immunofluorescence and Western blot. Then detected the alcohol preference and locomotor activity of mice with the open field test, conditioned place preference experiment, and loss of righting reflex test. Western blot analysis was used to identify neuron F-actin/G-actin ratio. Using Golgi staining, the morphology of dendritic spines was identified. Results In the alcohol two-bottle choice model, the expression of ICAM5 in the hippocampus of mice in the alcohol group was considerably lower than the control group. The specific expression of ICAM5 in the hippocampus of mice was observed by fluorescence microscopy. In the open field experiment, the staying time and moving distance of AAV-ICAM5 group were significantly increased compared with the control group (P < 0.01). In the CPP experiment, the residence time of AAV-ICAM5 mice in the alcohol-paired compartment was significantly lower than that of control mice (P < 0.001). In the lose the righting reflex experiment, overexpression of ICAM5 significantly reduced the sedation latency (P < 0.01), but significantly shortened the duration of sedation (P < 0.001). Compared with the control group, the ratio of F-actin/G-actin in hippocampus was significantly increased after drinking (P < 0.01). But after ICAM5 overexpression, the F-actin/G-actin ratio was significantly decreased (P < 0.001). Compared with the control group, the density of dendritic spines in hippocampal CA1 region was increased after drinking alcohol (P < 0.001), but the density of dendritic spines in AAV-ICAM5+EtOH group was significantly decreased (P < 0.01). Conclusions ICAM5 may modulated the expression of cytoskeleton protein to change structural plasticity of dendritic spines, which contribute to alcohol drinking and locomotor behaviors change of mice.
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Combined with systematic pharmacology and metabonomics to explore the mechanism of baicalein in the treatment of hyperuricemia

liangjingzhen, gaoyingjie, yewenqian, weibingyan, cheenzhaoyang, yangfan

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Objective to explore the effect and mechanism of baicalein in the treatment of hyperuricemia. Methods the mouse model of hyperuricemia was established by yeast extract combined with potassium oxazinate. The effect and potential mechanism of baicalein in the treatment of hyperuricemia were studied by biochemical indexes, pathological changes, non-target metabonomics and network pharmacology. Results baicalein could reduce the contents of uric acid, creatinine and blood urea nitrogen, reduce the inflammatory injury of renal tissue, up-regulate the expression level of uric acid excretion protein and down-regulate the expression level of uric acid reabsorption protein. Nine disease-related targets such as BCL2, SIRT1 and XDH were screened by network pharmacology. Six key metabolic pathways including nicotinic acid and nicotinamide metabolism, caffeine metabolism and purine metabolism were screened by metabonomics analysis. Conclusion baicalein can treat hyperuricemia and reduce renal injury, and its mechanism may be related to the metabolic pathways of nicotinic acid and nicotinamide regulated by SIRT1 and quinolinate.
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Progress in the use of gene editing technologies in the research of immunodeficiency animal models

mayunhui, wangxiaotang, gaojiping, songguohua

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Immunodeficiency animal models play an important role in preclinical research and are important experimental tools in modern biomedical research, widely used in immunology, genetics, oncology and microbiology, and other research fields. Gene editing is a technology of targeted modification of biological genomes, from emergence to application, it has greatly promoted the development of biomedical research. Gene editing technology mainly includes homing endonucleases (HEs), zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9 systems. At present, researchers has used these technology to establish a variety type of immune deficiency animal models, each with its own advantages and limitation. In recent years, a large number of studies have confirmed that the human immunodeficiency animal model can accurately simulate the function of cancer cells, drugs and immune system in human body, and can better simulate human diseases, and is widely used in the study of human immunobiology and the potential mechanism of human complex diseases. In this paper, we reviewed the progress in the research and application of gene editing technology in the construction of immune deficiency animal models, discussed the problems and optimization strategies of gene editing technology in the preparation of immune deficiency animal models in depth, and prospected its future development prospects, in order to provide references for researchers to select and build immune deficiency animal models.
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Extreme animal protectionism and experimental animal welfare ethics

Tian Xuemei, Sachiko Kubo, Wu Yingjie, Su Tao

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Experimental animals, as "living reagents", promote the development of life science and medicine, and also make the public pay more attention to the welfare ethics of experimental animals. Experimental animal welfare ethics is not only an important part of animal protection, but also plays an important role in protecting the rights and interests of experimental animals and promoting the rationality and morality of scientific research. However, in the evolution of animal protectionism, some extreme theory such as extreme animal protectionism had also emerged. This article summarized the main theories of animal protectionism and the development of animal welfare ethics and analyzed the main problems of extreme animal protectionism. This article expounds the present situation and problems of experimental animal welfare ethics and puts forward some suggestions to promote the practice of experimental animal welfare ethics, in order to provide references for our country's experimental animal welfare ethics practice and system construction.
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Comparative studies of transcriptomics in two murine liver fibrosis models induced by hepatotoxic chemicals

YAN Ruanyu, WU Hongyu, HUANG Kai, SUN Xin, XUE Jingbo, TAO Yanyan, LIU Chenghai, PENG Yuan

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Objective To compare the transcriptomic differences between carbon tetrachloride (CCl4)-induced and Diethyl 1,4-dihydro-2,4,6-trimethyl-3,5-pyridinedicarboxylate (DDC) diet-induced mouse models of liver fibrosis in order to provide a framework for future research utilizing mouse liver fibrosis models. Methods Mouse models of liver fibrosis were induced by 10%CCl4 (2ml/kg) injection and 0.1%DDC diet, respectively. After the 4 -week induction, the serum levels of ALT, AST, and TBil were detected. HE and Sirius red staining were observed to analyze the hepatic inflammation and collagen deposition. Jamall's method was used to evaluate the hydroxyproline (Hyp) content in the liver tissues. Hepatic tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) were measured by the elisa kits. Total RNA was extracted from murine liver tissues for RNA sequencing (RNA-Seq). The differentially expressed genes of the two models were analyzed by R software and then GO and KEGG enrichment were analyzed. Then, cwas used to verify the significantly different genes. Results Compared with normal mice, the serum levels of ALT, AST, TBil and the expressions of hepatic TNF-α, IL-6 and IL-1β were significantly increased in mice received CCl4-induction and DDC diet mice respectively, while the serum level of Alb was decreased. Pathological staining showed that the structures of liver tissues were destroyed and a large number of hepatocytes around the central vein were hyalinized and necrotic in CCl4-treated mice. In DDC diet mice, a lot of porphyrins were deposited in the liver and a large number of inflammatory cells were infiltrated in the portal area and the bile duct. Different degrees of collagen deposition were observed in the liver tissues of the two model mice. Different genes(DEGs) of CCl4-treated and DDC-diet mice were screened according to a filter (|logFC|> 2 times and P < 0.05). 1820 and 2373 DEGs in CCl4-treated and DDC-diet were analyzed, including 1302 and 1978 up-regulated genes , 518 and 395 down-regulated genes, respectively. GO annotation showed that the two models had important functions in molecular function (MF), biological process (BP) and cell component (CC). KEGG analysis showed that 22 and 29 signaling pathways were activated in CCl4-induction and DDC diet model, respectively. Among these, 16 signaling pathways such as extracellular matrix receptor interaction, cell cycle, protein digestion and absorption, focal adhesion, and PI3K-Akt were significantly enriched in the two models (P < 0.05). Cluster analysis showed that Mup11, Mup15, Mup17, Mup1 were significantly down-regulated in both models, which were identified by RT-qPCR (P < 0.05). Conclusions This study reported and compared the RNA-Seq transcriptomic characteristics of liver fibrosis models induced by CCl4-induction and DDC-diet, respectively. By observing the locations of gene expression and the pathways regulated by the genes, an example was established for the subsequent selection of animal models to study the pathogenesis and treatment of liver fibrosis.
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Mechanism of Juanxiao decoction regulating type 3 innate lymphoid cells in treating obese asthmatic mice

TIAN Min ping, ZHANG Qing yuan, XIANG Shuang di, CHENG Ling ling, SUN Peng, XUE Han-rong

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Objective: Explore the mechanism of Juanxiao Decoction in regulating the type 3 innate lymphoid cells (ILC3s) in the treatment of obese asthma. Method: Sixty male BALB/C mice were randomly divided into a normal group, a model group (High Fat Diet+OVA), and Juanxiao Decoction groups (low-, middle-, and high doses of 8.5, 17, and 34 g?kg_-1^{), dexamethasone group (}1 mg?kg_-1^{), with10 in each group.} Except for the normal group, other groups were fed high-fat diet for 12 weeks, and OVA sensitization and atomization inhalation were used to establish an obesity asthma model. From the first atomization, the low-, medium-, and high dose groups of Juanxiao decoction and the dexamethasone group were given corresponding drugs by gavage, while the normal group and model group were given equal amounts of saline by gavage for 7 days. Observing the state of each group of mice and changes in typical symptoms of obesity asthma. Within 24 hours after the last challenge, a fully automated biochemical analyzer was used to detect the four items of blood lipids and the classified count of inflammatory cells in alveolar lavage fluid (BALF). Hematoxylin- eosin (HE) staining was used to observe the morphological changes of lung tissue and abdominal fat in mice. Enzyme linked immunosorbent assay (ELISA) for detecting levels of immunoglobulin E (IgE) in BALF and serum of mice, as well as the levels of interleukin-1β(IL-1β)、Interleukin-13(IL-13)and mouse thymus activation regulating chemokine (CCL17) in lung tissue. The levels of IL-17A₊^{ILC3 and IL-22}_⁺ILC3 cells in lung tissue and peripheral blood of mice were analyzed by flow cytometry. Western blot was used to detect the expression of P-STAT3 protein in lung tissue. Results: Compared with the model group, the general state and typical obese asthma symptoms in the middle and high dose groups of Juanxiao decoction were obviously improved. Pathological staining showed that the airway inflammatory cells infiltrated and the airway wall thickened in the model group. However, compared with the model group, the lung inflammation in dexamethasone group and Juanxiao decoction group was improved, especially in the middle and high dose groups. The levels of IL-1β, IL-17A_⁺ILC3, IL-13 and CCL17 in lung tissue of mice in model group were higher than those in normal group (p＜0.05), while the proportion of IL-22_＋^ILC3 and the expression of P-STAT3 were significantly lower than those in normal group(p <0.01,p＜0.05). After the intervention of middle and high dose of Juanxiao decoction, the levels of IL-1β, 17A₊^ILC3, IL-13 and CCL17 decreased significantly(p＜0.05,p＜0.01,p＜0.001)and the the proportion of IL-22_＋^ILC3 and the expression of P-STAT3 increased significantly(p＜0.05,p＜0.01,p＜0.001).Conclusion: Juanxiao decoction can alleviate lung inflammation and allergic reaction in obese asthmatic mice, and improve the inflammatory environment of the body, and its mechanism may be related to regulating the secretion of cytokines by ILC3.
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Adra1a regulates LPS-induced inflammation in primary hepatocytes of Lbp-/-mice

MI Chuanliang, FU Bin, LI Sidi, CHEN Zhida, GUO Zhongkun, WANG Kezhou

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Objective To Explore the Adra1a regulation of inflammatory response in primary hepatocytes of LBP knockout mice (Lbp-/-) induced by LPS. Method Using two-step perfusion method to extract primary hepatocytes and constracting an primary hepatocyte inflammation model from WT group and Lbp-/- group. Inhibition of Adra1a expression in Lbp-/- mouse primary hepatocytes by adding inhibitor prazosin and transfecting siRNA .The inhibitor method divided the cells into control group (blank control), LPS group (LPS stimulation), and inhibitor piperazine group (LPS stimulation added after 1 hour of prazosin intervention). The method of siRNA transfection divided primary hepatocytes into four groups: control group (blank control), LPS group (LPS stimulation for 12 hours), negative control group (si-NC interference for 12 hours followed by LPS stimulation for 12 hours), and interference group (si-adra1a interference for 12 hours followed by LPS stimulation for 12 hours). The primary hepatocytes of WT mice were divided into two groups: the control group (blank control) and the LPS group (LPS stimulation for 12 hours). This study used WT type and Lbp-/- type mouse primary liver cells as the research subjects to verify the changes of Adra1a under LPS stimulation using Western blot method. Experimental methods such as CCK8, qPCR, and Western blot were used to verify the improvement of inflammation and survival rate of primary liver cells in Lbp-/- mice treated with prazosin and si-adra1a. Results under LPS stimulation, the expression of Adra1a protein in primary hepatocytes of Lbp-/- mice was significantly increased (P < 0.01), while the wild-type did not change; The cell survival rate of the inhibitor prazosin group and the interference group was significantly increased (P < 0.01; P < 0.05); In the inhibitor prazosin group and interference group, the expression of inflammatory factors TNF-α、IL-1β significantly decreased (P < 0.01). The expression levels of proteins p-p38, p-ERK, and p-JNK related to cell damage and inflammation were also significantly reduced (P < 0.01). Conclusion upregulation of Adra1a expression in Lbp -/- mouse primary liver cells induced by LPS compensates for the role of lipopolysaccharide binding protein (LBP) in conducting injury and inflammatory signals, inhibiting the expression of Adra1a gene can significantly reduce the occurrence of inflammation and cell damage in primary liver cells.
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Study on the effect of radiation on mouse salivary gland tissue damage based on NLRP3

wuyuqi, huangguilin, xiaolijun, zhangmin, zhangnini

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Objective: To study the effects of radiation on the morphology, function, and NLRP3 expression of mouse salivary gland tissue, and provide new ideas for repairing radiation-induced damage to salivary gland tissue. Methods: To establish a mouse model of radiation-induced submandibular gland injury and record the weight of drinking water. Salivary flow rate was detected, HE staining was used to observe the submandibular gland injury and Immunohistochemistry and real-time PCR were used to detect the expression of NLRP3 and Caspase-1 in the radiation-induced submandibular gland injury of mice at 1, 3, 7 and 14 days after radiation. Results: With the accumulation of time, the amount of water consumed by the radiation group mice gradually increased, the salivary flow rate decreased, and inflammatory cells in the submandibular gland continued to increase. acinar cells gradually showed lesions such as nuclear pyknosis and vacuolization. At 7 and 14 days after radiation, the expression levels of NLRP3 and Caspase-1 protein and gene in the radiation group were significantly higher than those in the normal group (P < 0.05). Conclusion: Radiation can induce damage to mouse submandibular gland tissue and activate NLRP3 inflammasome to increase its expression level.
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Study on the efficacy of 1470 nm semiconductor laser therapy instrument for canine prostatectomy

HUANG Heng, PAN Yongming, HUANG Junjie, ZHANG Hui, YU Chen, CHEN Minli, XU Qingfeng, ZHENG Guo

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Objective To evaluate the effectiveness of 1470 nm semiconductor laser therapeutic instrument (referred to as curestar therapeutic instrument) for prostatectomy in Beagle dogs. Method 28 adult male Beagle dogs were randomly divided into 3 groups: sham group (n=3), experimental group (n=15), and control group (n=10). The experimental group was further divided into 3 subgroups: 120 W/50 W, 150 W/50 W, and 160 W/50 W for vaporization cutting/coagulation hemostasis, while the control group was divided into 2 subgroups: 120 W/50 W and 150 W/50 W. 5 in each subgroup. Both the experimental group and the control group underwent canine prostatectomy through the entrance of the bladder neck under electrocision. The operational suitability and effectiveness of the product during surgery were observed. After the operation, the general condition of the dogs was observed, and blood biochemical and hematological indicators were measured at before, immediately, 3 days, 7 days, and 28 days after operation. At 1h and 4 weeks after surgery, B-ultrasound and electric resection were performed under anesthesia to observe the condition of the urethra and prostate, and the prostatic tissue was subjected to H&E staining for pathological observation. The thickness of the coagulation layer at 1h after the operation and the repair of the urethelial epithelium at 4 weeks were analyzed. Results During the operation, the experimental and control groups had good operability and showed good vaporization cutting and coagulation hemostasis performance. After the operation, there were no significant effects on the general condition, blood biochemical, and hematological indicators of the dogs. Ultrasound showed that the urethral expansion was visible immediately after the operation, and the echo of the urethral epithelium was slightly enhanced. At 4 weeks, the prostate tissue was slightly low-echo with uniformly distributed small point-like echoes inside, and the capsule was linearly high-echo, consistent with the sham group. The measurement and analysis showed that the weight of the vaporized prostate tissue in the experimental and control groups was about 0.91-1.33 g, with a resection rate of 17.11-20.27 %. And as the power of vaporization cutting increases, the laser emission time gradually decreases, while the vaporization cutting speed and efficiency both increase.However, there was no significant difference between the experimental and control groups (P>0.05). Under the electrocision microscope, a burn-like change was observed in the surgical wounds of the prostate urethra in the experimental group and the control group at 1 h after surgery, and the boundary between the wound and the normal urothelium was clearly visible. At 4 weeks, the urothelium of the prostate had been repaired and flattened, and the boundary with the surrounding normal urothelium was blurred. Similarly, pathological observations also showed that at 1 h after surgery, both the experimental and control groups showed significant damage to the prostate urethral orifice, with a small amount of carbonization and coagulative necrosis on the surface of the wound, a small amount of inflammatory cell infiltration, and a coagulation layer thickness of about 0.4 mm. At 4 weeks, the prostate urethral morphology of the sham group was normal, while the experimental group and the control group both showed new epithelial growth covering the wound, with uniform thickness and no coagulative necrosis tissue attached to the wound. Mild inflammatory reaction was still present in the surrounding area, fibroblast proliferation was obvious, and stromal and epithelial cell proliferation was visible in the surrounding prostate, some of which showed squamous metaplasia. The prostate capsule was intact and the morphology of the surrounding nerves and blood vessels was normal. Conclusion The curestar therapy instrument is effective for prostatectomy in Beagle dogs, with good vaporization cutting and coagulation hemostasis performance, and there was no significant difference in postoperative physiological indicators compared to the sham group.
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Characteristics燼nd營nnovations爄n燛ducation爋f爈aboratory animal science 爄n燭raditional燙hinese燤edicine燙olleges燼nd燯niversities

ZHANG Min, LIU Yanyun, GU Xin, TIAN Daizhi, YANG Yang

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Laboratory animal science plays燼燾rucial爎ole爄n爐he爀ducation燼nd爐raining爋f爉edical爏tudents.燭o爀nsure爐he爀ffective爐eaching爋f爈aboratory animal science爄n燙hinese爉edicine爄nstitutions,爄t爄s爄mperative爐o爄ncorporate爐he爌rinciples爋f燙hinese爉edicine燾haracteristics燼nd爄nnovation.燭his爌edagogical燼pproach燼ims爐o爀nrich爐raditional燾ultural爈iteracy爓ithin燙hinese爉edicine燼nd爁oster爏tudents'燾apacities爁or爄nnovation燼nd燾ritical爐hinking.燭he燿aily爐eaching爋f爈aboratory animal science爏hould爀ncompass爒arious爀ducational燼spects,爄ncluding爄nheritance爀ducation,爀thical爀ducation,爉igration爀ducation,爀xploration爀ducation,爄nspiration爀ducation,燼nd爀xtension爀ducation.燘y爄ntegrating爐hese燾omponents燼nd爀mphasizing爐he爄ntersections爋f燙hinese爉edicine爓ith爈aboratory animal science,爄mportant燼reas爏uch燼s燼nimal爓elfare爄n燙hinese爉edicine爎esearch,爐he燼pplication爋f燙hinese爉edicine爐heories爄n燼nimal爀xperiments,燾onsiderations爁or燼nimal爉odels爄n燙hinese爉edicine,燼nd爐he爏election爋f爀xperimental燼nimal爏pecies,燾an燽e燼ddressed.燗dditionally,爐his燼pproach爌rovides爂uidance爁or爊ew燿evelopment燿irections燼nd燾ultivates爏tudents'爏cientific爎esearch,爄nnovation,燼nd爌ractical燼bilities爀ffectively.燭he爌rimary爋bjective爋f爐his爌rogram爄s爐o爊urture爏cientific爄nnovation燼nd爌ractical燾ompetence燼mong爏tudents爓hile爌rioritizing爐he爏uperior爍uality爋f爄nnovative爀ducation爓ithin爐he燾ontext爋f燙hinese爉edicine.燬uch爀fforts爋ffer燼爏olid爁oundation爁or燼dvancing爄ndigenous爉edical爌ractices燼nd爊urturing燼爊ew爂eneration爋f爌rofessionals燿edicated爐o爌reserving爐raditional爉edicine.
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Effects of acute sleep deprivation on behavior and synaptic biomarker expression in rats at different times

zhangshibin, wang lu, wang chu, guo pengcheng, yan xusheng, huo dongsheng, yang zhanjun, wang yanguo, jia jianxin

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Objective: To investigate the effects of acute sleep deprivation on the behavior and synaptic protein expression in rats at different times. Methods: Seventy healthy male Wistar rats were randomly divided into 7 groups, control group, different time sleep deprivation groups (24 hour, 48 hour, 72 hour, 96 hour, 120 hour and 144 hour). The sleep deprivation rat model was established by modified multi-platform water environment sleep deprivation method. Spatial learning and memory was detected by Morris water maze, anxiety was detected by open field test, and the morphology and quantity of hippocampal neurons were observed by Nissl staining. Western blotting and Real-time PCR determined the expressions of synaptophysin (SYN), post-synaptic density protein-95 (PSD-95) and brain-derived neurotrophic factor (BDNF) in rats. Results: Compared with the control group, the numbers of standing and modification all significantly increased accompanied by the prolongation of sleep deprivation time (P<0.05). The escape latency and path length significantly increased in rats in the 120 h and 144 h groups (P<0.05), while the number of crossing platforms and the percentage of target quadrant time all significantly decreased (P<0.01), and were negatively correlated with sleep deprivation time. The expression levels of BDNF, SYN and PSD-95 all significantly decreased accompanied by the prolongation of sleep deprivation time (P<0.01), and was negatively correlated with sleep deprivation time. Conclusion: With the increase of sleep deprivation time, cognitive dysfunction and anxiety gradually deteriorated, which may be related to decrease in expression level of synaptic biomarkers.
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Exploring the mechanism of Inonotus obliquus extract in treating Crohn?s disease based on proteomics

han lihua, Zhao Xiaoru, Peng Lili, Hao Miao, Yuan Hongxia

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Objectives To investigate the effect of Inonotus obliquus extract on Crohn's disease and its mechanism by proteomics technology. Methods Crohn's disease (CD) model was induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). A total of 48 SD male rats were randomized into control, model, mesalazine (225 mg/kg), and low-dose (200 mg/kg), medium-dose (400 mg/kg), high-dose (800 mg/kg) Inonotus obliquus groups. The disease activity index (DAI) score and the colonic mucosal injury index (CMDI) score were assessed after one week of drug intervention. HE staining was used to observe the histopathological changes in the colon, and ELISA was used to detect the levels of IL-1β, IL-6, and TNF-α in the blood. Proteins were extracted from the colonic tissues of the control group, model group, and Inonotus obliquus extract high-dose group, and bioinformatics analysis was performed for the proteins identified by quantitative proteomics. Finally, WB and RT-PCR were employed to verify the key proteins. Results The Inonotus obliquus extract improved the symptoms of colitis induced by TNBS in rat, and decrease the levels of IL-1β, IL-6, and TNF-α in the blood. Proteomic tests showed that there were 199 differentially expressed proteins (DEPs) between the Inonotus obliquus extract high-dose group and the model group, of which 63 DEPs were related to CD. Bioinformatics analysis showed that these 63 DEPs were mainly involved in NOD-like receptor signaling pathway, calcium signaling pathway, necroptosis, and other pathways. Consistent with proteomic results, expressions of Vdac1 and Trpv2 were confirmed by WB and RT-PCR in colon tissue. Conclusion Inonotus obliquus extract may regulate NOD-like receptor signaling pathway, calcium signaling pathway, and necroptosis by interfering with the expression of Vdac1 and Trpv2, so as to achieve the effect of treating CD.
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The research progress of traditional Chinese medicine treatment of breast cancer depression and co -disease

PENG Mengwei, WU Yaosong, LIU Gaoyuan, KUANG Luoyi, LUO Zilong, CHEN Yulong, LIU Yan

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Pathological mood changes, mainly depression, occurring during the diagnosis and treatment of breast cancer are clinically called breast cancer related depression(BCRD). A large number of epidemiology and clinical studies have confirmed that BCRD is complex， difficult to treat,and has a poor prognosis. Most of the existing clinical treatments are postoperative chemotherapy for breast cancer and antidepressant drugs, which treats breast cancer and depression as two independent diseases,with many defects such as low efficiency and strong adverse reactions. Traditional Chinese medicine（TCM）has a unique value in the prevention and treatment of breast cancer-related depression by virtue of its significant advantage of regulating multiple pathways and targets with its multiple components at the same time.In this paper,we review the mechanism of BRCD and the therapeutic mechanism of TCM from the aspects of neurological disorders, inflammatory immune response, and intestinal flora disorders, with a view to providing certain references for the clinical application and research of TCM in the treatment of BCRD.
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Study on the therapeutic effect of Qingre Jiedu formula on gout mice based on immune inflammation and intestinal microecology

hexianshun, linkun, weiyurou, tianjiaqing, jiangyulai, weitengfei, lintianye, hemincong, weiqiushi

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Objective: To explore the anti-inflammatory effect of Qingre Jiedu（QRJD） Formula on gout mice and its effect on gut microbiota.Method: Forty 20-22 g C57BL/6 were divided into Control group（CON）, model group（MOD）, allopurinol group（Allo）, QRJD Formula group（QRJD）, and ig 10 g/ 0.1ml carboxymethyl cellulose in blank group every morning from 1 to 28 days. Hyperuricemia mouse model was prepared by potassium oxyazinic acid (500 mg/kg) + yeast extract (10 g/kg) suspension intragaically. On the 29th day, 80ul sterile carboxymethyl cellulose was injected into the left ankle of mice in the CON group under isoflurane anesthesia, and gouty arthritis model was prepared by injecting the same amount of sodium urate solution into the left ankle of mice in the other groups. At the same time, each group was treated with corresponding drugs every day. On the 35th day, samples were taken from mice who had been fasting for 6 hours without water. Blood indexes such as uric acid, creatinine and urea nitrogen were detected. Hematoxylin-eosin staining was performed on ankle joints. The cecum contents of mice were collected and the changes of gut microbiota were detected by 16S rDNA high-throughput sequencing method. Results: ① After 7 days of treatment, compared with MOD group, QRJD formula can effectively reduce the concentrations of blood uric acid (P < 0.001), creatinine (P < 0.001) and urea nitrogen (P < 0.05), and effectively protect renal function; ② The pathological results indicated that compared with the MOD group, HE staining showed that the synovial hyperplasia and inflammatory cell infiltration were reduced in the QRJD formula group after treatment. It was found that the cartilage arrangement of the compound was more orderly than before, the cartilage destruction was less than that of the MOD group, and no matrix loss was observed.③ The immunohistochemical results of ankle joint indicated that IL-10 and TGF-β1 were not significantly increased in CON group and MOD group; Compared with MOD group, the expressions of IL-10 and TGF-β1 of QRJD recipe were increased (P < 0.05).④ In terms of biodiversity, there was no significant difference in α diversity among the four groups (P < 0.05), while β-diversity was found to be more similar to the CON group (P=0.001).⑤Compared to the CON group, the MOD group exhibited increased abundances (P < 0.05) of Ruminococcaceae spp., Dubosiella sp., Tyzzerella sp., Ileibacterium sp.and Bacteroidales spp.In contrast to the MOD group, the QRJD group showed elevated abundances (P < 0.05) of Lactobacillus sp., Ligilactobacillus sp., and Bacteroides sp. Furthermore, the interaction network of the gut microbiota indicated mutual interactions among these microorganisms.⑥In the correlation analysis between gut microbiota and renal function, as well as anti-inflammatory factors, it was observed that the relative abundance of Dubosiella sp., Tyzzerella sp.and Bacteroidales spp. was significantly positively correlated with SUA and SCR (P < 0.05). On the other hand, Lactobacillus sp., Ligilactobacillus sp., and Mitochondria spp. exhibited a positive correlation with anti-inflammatory factors IL-10 and TGF-β1, with a more significant association observed with TGF-β1 (P < 0.05).⑦COG function prediction suggested that the functions of QRJD formula group were concentrated on inorganic ion transport and metabolism, carbohydrate transport and metabolism, etc. Conclusions: QRJD effectively modulates immune inflammation and gut microbiota dysbiosis, thereby treating gout. It is hypothesized that its mechanism of gout prevention and treatment involves the regulation of gut microbiota diversity and abundance, as well as the control of the abundance of differential bacterial species such as Ruminococcaceae spp., Dubosiella sp.and Lactobacillus sp. to achieve the goal of gout therapy.
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Research progress on the involvement of macrophages in myocardial ischemia reperfusion injury mediated by NF-κB signaling pathway

lishilong, wanghe

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Myocardial ischemia reperfusion injury（MIRI）has become one of the most serious complications affecting the clinical outcome of patients with cardiovascular diseases. The immune inflammatory response of macrophages is closely related to the occurrence and development of MIRI.Many studies have shown that the NF-κB signaling pathway can participate in MIRI regulation by influencing the polarization and inflammatory state of macrophages, pyrodeath, infiltration and other functions, and is a potential target for MIRI therapy.Therefore, this article will review the research progress of NF-κB signaling pathway between macrophage function and MIRI regulation.
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Comparison of liver and adrenal orthotopic transplantation models of neuroblastoma

chen Hongxia, Tan Zhigang, Lin Huiran, Feng Luping, Zheng Chuya, Liao Wenfeng, Zeng Rufeng, Liu Jinxin, Zhuo Zhenjian

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【Abstract】Objective The Neuroblastoma (NB) liver transplantation model was established and compared with the adrenal orthotopic transplantation model to explore the characteristics of the model.. Methods 5×105 (SK-N-SH) cells was implanted into the left lobe of mouse liver along the long axis of liver lobe with a micro-injection needle. The growth, metastasis, expression of related genes and histopathological changes of tumor were detected after the modeling. Results The tumor formation rate of mice inoculated with tumor cells reached 100% after 21 days, and the tumor growth, metastasis, related gene expression and pathological characteristics were obvious. Conclusion In this paper, neuroblastoma liver transplantation model was successfully constructed by a relatively simple surgical modeling method, which provides more ideal model choices for future scientific experiments of NB researchers.
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A comparative study of two methods for the clearance of macrophages from the rat endometrium

XIA Liangjun, CUI Chuting, LI Junwei, CHENG Jie, XIA Youbing

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Objectives A comparison of different administration routes of clodronate disodium liposomes to determine which is the most effective method of rat endometrial macrophage clearance. Methods Female 8-week-old SD rats were randomly divided into the unilateral control group (injected 100μL PBS liposomes into the left uterine cavity), unilateral clearance group (injected 100μL clodronate liposomes into the right uterine cavity), bilateral control group (injected 100 μL PBS liposomes into the bilateral intrauterine), bilateral clearance group (injected 100 μL clodronate liposomes into the bilateral intrauterine), and whole-body control group (injected 500 μL PBS liposomes into the caudal vein ) and whole-body clearance group (injected 500 μL clodronate liposomes into the caudal vein). Haematoxylin and eosin (H&E) staining was used to observe the morphology and structure of uterine and ovarian tissues, immunohistochemistry was used to observe the positive presence of macrophages in uterine and ovarian tissues, and flow cytometry was used to detect changes of the macrophages cell counts in uterine and ovarian tissues. Results There was no significant difference in the structure and morphology of the uterus and ovary among all groups. Immunohistochemical staining showed that compared to the control group, the positive expression of uterine macrophages in unilateral and bilateral clearance groups was significantly decreased (P<0.001), and had no effect on the accumulation of macrophages in the ovary (P>0.05). The numbers of macrophages in both uterine and ovarian tissues decreased of whole-body clearance group (P<0.01, P<0.01). Compared with unilateral and bilateral clearance groups, whole-body clearance group had a greater effect on the positive expression of macrophages in ovarian tissues (P<0.05). Flow cytometry showed that compared with the control group, the percentage of macrophages in the uterine tissue was significantly reduced in each clearance group (P<0.001, P<0.001, P<0.05). Compared to the whole-body clearance group, the clearance effect of endometrial macrophages was better in the unilateral and bilateral clearance groups (P<0.05，P<0.05). In addition, the number of macrophages in ovarian tissue decreased in all clearance groups compared to the control group. The decrease in macrophage number was most pronounced in the whole-body clearance group (P<0.05), and there was no statistically significant difference between the unilateral and bilateral clearance groups compared with the control group (P>0.05).
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Effect of Alzheimer"s disease on auditory function in APP/PS1 transgenic mice

GUI Fei, SONG Dan-dan, WANG Hai-yan, SUN Xiao-pin, Yang Lei

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Objective To explore the hearing and cochlear himorphological changes in APP/PS1 transgenic mice during the process of AD, and to determine whether the occurrence and development of AD will affect their hearing function. Methods APP/PS1 transgenic mice and littermates wild type mice were selected at the age of 4, 8 and 12 months. The changes of auditory function in APP/PS1 transgenic mice at different months were detected by immunofluorescence staining and auditory brainstem response (ABR). Results Compared with the control group, the number of Aβ plaques in the hippocampus of 4-month-old APP/PS1 transgenic mice was significantly increased, indicating that AD symptoms were already present at this time. At the age of 4, 8 and 12 months, there was no significant difference in the hearing threshold between APP/PS1 transgenic mice and wild type mice. The histomorphological examination of the cochlea showed that there was no significant difference in the key cells of the inner ear such as cochlear hair cells and spiral ganglion between the two groups of mice. ABR test results of APP/PS1 transgenic mice and littermate wild-type mice showed that the hearing threshold of both mice increased significantly with age, both of them showed age-related hearing loss. Conclusion There is age-related hearing loss in APP/PS1 transgenic mice, but the occurrence and development of AD has little effect on the hearing function of APP/PS1 transgenic mice.
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Improvement of Depressive-like Behavior in Hemerocallis citrina Baroni Using a Zebrafish Model

Zheng Hanwen, Wang ZiHang, Liu Xinyue, Yu Jiawei, Alberto Carlos Pires Dias, Wang Qiong

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Abstract OBJECTIVE: This study aimed to investigate the potential positive effects of the alcoholic extract of Hemerocallis citrina Baroni on depressive behaviors in zebrafish larvae (Danio rerio) induced by reserpine. METHODS: Zebrafish larvae were divided into various groups, which included a control group, a model group, a model group with fluoxetine (a positive drug), a model group with a low dose of HCE (1.5 mg/L), a model group with a medium dose of HCE (3 mg/L), and a model group with a high dose of HCE (4.5 mg/L). Depression-like behaviors were analyzed using sound and light stimulation. RT-qPCR was utilized to investigate the effects of the alcoholic extracts of HCE on depression-related astrocyte markers (GFAP, C3, C4B, EMP-1, S100α-10) as well as the neurotrophic factor (BDNF) and its receptor genes (P75, TrkB). RESULTS: In comparison to the control group, the model group demonstrated significantly shorter movement distance and reduced movement time under sound and light stimulation (P<0.05, P<0.0001). Following the administration of the alcoholic extract of HCE, zebrafish larvae exhibited significantly heightened sensitivity to light and sound stimulation when compared to the model group (P<0.05, P<0.01). Astrocyte marker genes were up-regulated in the zebrafish brain in the model group compared to the control group (P<0.01). However, when the model group was administered with the alcoholic extract, the expression of astrocyte markers was significantly down-regulated compared to the model group (P<0.01). Neurotrophic factor and its receptor genes (BDNF, P75, TrkB) were down-regulated in the zebrafish brain in the model group compared to the control group (P<0.01). However, in the group administered with Brassica campestris alcoholic extract, the expression of BDNF, P75, and TrkB was significantly up-regulated compared to the model group (P<0.001). These findings suggest that the alcoholic extract of HCE suppresses the inflammatory response caused by astrocyte activation and promotes the production of neurotrophic factors and their receptor genes, thereby exerting an ameliorative effect on depression. CONCLUSION: Alcoholic extracts of HCE can improve depression-like behavioral changes induced by reserpine in zebrafish larvae . They reduce the expression of astrocyte markers in the zebrafish brain and promote the production of neurotrophic factors and their receptor genes, playing an antidepressant role.
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Establishment and validation of a liver injury model in mice induced by chronic low-dose exposure to atrazine

ZHU Yu, SU Ying-shi, HE Bao-guo, LIU Xi, QIN Lei

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Objective To establish a model of long-term atrazine (ATR)-induced liver injury in mice and to evaluate the hepatotoxic effects induced by ATR. Methods C57BL/6-N male mice were randomly divided into drinking control group, 1.5mg/L and 150mg/L ATR dose groups. Serum liver function biochemical indexes and inflammatory factors were detected after 35 and 63 d, hepatosomatic ratio was calculated, and histopathology and ultrastructure of the liver were observed. The levels of lipid peroxidation and antioxidant capacity, the activities of major phase I metabolic enzymes and phase II detoxification enzymes, the expression of related proteins in liver tissues were detected. Results Compared with control group, AST/ALT ratio, pro-inflammatory factors CCL2, TNF-α and IL-6, H2O2 content and the activities of metabolic enzymes NCR, CYPb5 and UDPGT in ATR groups had significant changes (P < 0.05). In 150mg/L ATR group, GGT content, MDA level of peroxide and CYP1A2 expression were significantly increased (P < 0.01), while GSH content was significantly decreased (P < 0.05). Moreover, hepatocyte injury and mitochondrial vacuolation were more serious. Conclusions In a mouse model of low-dose ATR- liver injury, both 1.5 mg/L and 150 mg/L atrazine exposure induced liver injury in mice, with 150 mg/L ATR inducing greater metabolic toxicity in the liver after 63 d.
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Research progress on roles of ferroptosis in chemotherapy resistance of leukemia

anlijuan, haozheng

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Chemotherapy resistance in leukemia is an urgent clinical therapeutic challenge. Ferroptosis is a unique mode of cell death driven by iron-dependent phospholipid peroxidation. Since leukemia is characterised by increased oxidative stress and iron overload, it can be hypothesised that leukemia cells are susceptible to ferroptosis, suggesting therapeutic potential. In recent years, ferroptosis has been extensively studied and used in the treatment of various types of leukemia. Several studies have demonstrated the association between the regulatory pathways of ferroptosis and the mechanisms of leukemia drug resistance. The induction of ferroptosis through different pathways can effectively reduce the resistance of various types of leukemia cells to chemotherapeutic drugs, and thus improve the clinical efficacy. This article summarizes the regulatory mechanisms of ferroptosis, analyzes in detail the association between oxidative stress pathways and iron metabolism pathways of ferroptosis and the mechanism of leukemia drug resistance, and compiles the experimental studies and clinical applications of ferroptosis in the treatment of various types of drug-resistant leukemias, with the aim of providing new ideas and directions for the study of ferroptosis and a new strategy to reverse chemotherapy resistance for leukemia patients in the future.
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Analysis on the Construction and Practice of Animal Biosafety Level-3(ABSL-3)Simulated Laboratory

Li Xiaoyan, Lu Xuancheng, Lu Shuangshuang, Wang Jiaqi, Kalibixiati Aimulajiang, Liu Mei, Liu Keliang, Zainawudong·Yushan

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ABSL-3 laboratory is Animal Biosafety Level-3 laboratory where highly pathogenic microorganisms can be done. In recent years, with the continuous emergence of emerging and re-emerging infectious diseases, high-level biosafety laboratories have played an increasingly important role in the hardware support of pathogenic mechanism, drug and vaccine research and development. The demand for ABSL-3 laboratories is increasing year by year. At the same time, there is also an increasing demand for personnel competence of working in ABSL-3 laboratories. The standardization of pre-service training has become an important guarantee to reduce the risk of personnel working in the ABSL-3 laboratory. The systematized, normalized and standardized training work needs to be carried out in a specific place. Simulated laboratory is the best place. Therefore, it is necessary to establish animal biosafety level-3 simulated laboratory and establish scientific and effective operating standards and mechanisms. This paper introduces the design, construction, operation and function of ABSL-3 simulated laboratory.
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Measurement and Comparative Analysis of Blood Physiological and Biochemical Indexes of Spontaneous Type 2 Diabetes Chinese Hamster in the SHANXI MU Colony

yangyuting, yangyichun, weibingyan, wujianqin, zhangruihu, chenzhaoyang

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Objective: Investigate the blood physiological and biochemical changes in the spontaneously developed type 2 diabetes mellitus (T2DM) animal model of the SHANXI MU colony inbred strain established by the research team in the preliminary stage. Methods: A cohort of spontaneously developed T2DM Chinese hamsters and normal hamsters, each comprising 10 individuals at 12 months of age, were selected for the study. Fasting blood samples were collected and analyzed using the Sysmex XT automated hematology analyzer and the Hitachi fully automatic biochemical analyzer to measure and statistically analyze 15 physiological parameters and 16 biochemical indicators in the blood. Results: Compared with the control group, WBC, RBC, PLT, HGB, ALT, AST, GLU, TC, TG and UA in diabetic group were significantly different(P < 0.05). Conclusion: The blood physiological and biochemical indexes of the Chinese hamster spontaneous T2DM model established by the research group in the early stage were detected, and the changes were in line with the trend of human T2DM incidence, which provided basic data for the application and application of the model.
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Progress in the optimization of CRISPR/Cas9 technology for the construction of genetically modified mouse models

wangyu, wuyong, liangchunnan, fanchangfa

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The rise of CRISPR/Cas9 technology has driven the development of various fields of life science. With the continuous deepening of its understanding, people have made multiple improvements and optimizations to adapt to different application scenarios. The optimization of CRISPR/Cas9 technology has also brought many breakthroughs in the production of genetically modified mouse models. This article briefly reviews the development process of CRISPR/Cas9 technology, and summarizes optimization strategies at the level of CRISPR/Cas9 element optimization, construction of conditional knockout/knockin gene modified mouse models, and delivery systems for CRISPR/Cas9 elements and HDR templates. It looks forward to the future development of this technology.
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The correlation of periodontitis and central nervous system inflammation in hypobaric hypoxia environment at plateau

chen lu, jiang xiao xia, zhu ling ling

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In recent years, with the increasing number of people active in the plateau and mountainous areas, high altitude exposure has become increasingly common, and many patients with underlying diseases are affected by the hypobaric hypoxia of the plateau environment, which further aggravates the disease process and even leads to cognitive disorders. Periodontitis is a common inflammatory disease that induces periodontal local inflammatory responses and even causes central nervous system inflammation. At high altitudes, the body suffers from decreased immunity and tissue hypoxia, which can promote the occurrence and development of periodontitis and may even increase the risk of periodontitis leading to central nervous system inflammation. With the deepening of plateau medicine research, the relationship between periodontitis and central nervous system inflammation in the hypobaric hypoxia of plateau environment has attracted more and more attention. This work reviews the research progress of periodontitis and central nervous system inflammation and discusses the correlation between periodontitis and central nervous system inflammation in the hypobaric hypoxia environment at plateau.
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Research progress of exercise on muscular atrophy based on mitochondrial quality Control

GUO Xiaojing, WANG Yan

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Skeletal muscle wasting refers to the loss of skeletal muscle mass and function. Mitochondrial quality control (MQC) is the basis for maintaining normal physiological mitochondrial function, which mainly involves the regulation of mitochondrial biogenesis, mitochondrial dynamics (fission/fusion) and mitophagy. It maintains muscle homeostasis by regulating the relative stability of mitochondrial shape, quantity and quality. As an economical and effective treatment for muscular atrophy, exercise intervention has been widely used, but the relationship between exercise intervention and MQC is not clear. This paper discusses the role of mitochondrial biogenesis, mitochondrial dynamics, and mitophagy in skeletal muscle atrophy and related molecular targets, and thoroughly analyzes the mechanism of MQC-mediated exercise in improving skeletal muscle atrophy caused by aging, disuse, and cancer cachexia, in order to provide theoretical guidance for exercise intervention in skeletal muscle atrophy.
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A comparative study of enzymatic digestion methods for the preparation of single cell suspensions from the lamina propria of the mouse intestine

zengqin, liangying, wangxinhui, yanlei, wangxiangpeng, yangjiayi, yurenhuan

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Objective To investigate the best digestion method for the preparation of single cell suspensions from mouse small intestinal lamina propria. Methods Ten mouse small intestines of uniform length were collected and randomly divided into two groups. Each group prepared the lamina propria single cell suspensions by enzymatic digestion with collagenase A or collagenase VIII, and compared the effects of these two enzymatic digestion methods on the cell yield, cell activity and cell surface antigen of the single cell suspensions, and then the single cell suspensions prepared by the superior enzymatic digestion method were subjected to flow assay. Results Compared to collagenase VIII-based enzymatic digestion, collagenase A-based enzymatic digestion resulted in higher cell yields (9.48 ?1.10) ?109 vs. (4.18 ?1.02) ?109, live cell proportions (86.36 ?3.32) % vs. (61.62 ?10.93) % and active CD45 cells (57.19 ?5.11) ) % vs (26.01?1.44) %, active CD3 cells (8.73?.89) % vs (4.52?.49) %, active CD4 cells (6.19?.09) % vs (3.22?.91) % and active B220 cells (15.06?.27) % vs (5.07?.20) %, providing high quality cells for subsequent flow assays. Conclusion The collagenase A-based enzymatic digestion method is more suitable for the preparation of single cell suspensions from the lamina propria of the mouse small intestine.
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Establishment of chimeric rabbits with FOXN1 gene knockout

liu tian ping, li gui ling, liu ke, chen bang zhu, wang gang, gu wei wang

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Objective The aim of this study was to establish F0 generation chimeric rabbits with FOXN1 gene knockout to explore methods for in vivo conservation of immunodeficient rabbits in a conventional housing environment. Methods Initially, the CRISPR/Cas9 technology was employed to inject the constructed sgRNA and Cas9 protein into a single cell of rabbit two-cell stage embryos to obtain chimeric embryos with FOXN1 gene editing. Subsequently, the embryos were transferred into surrogate does. Finally, the F0 generation offspring were genotyped using PCR and Sanger sequencing, and their growth and development in a conventional housing environment were observed. Results The PCR and Sanger sequencing results confirmed the successful establishment of chimeric rabbits with FOXN1 gene knockout. Observation revealed that the chimeras exhibited normal growth and development in a conventional environment without any immunodeficient phenotypes. Conclusion This study has initially established a chimeric rabbit model with FOXN1 gene knockout, which can grow and develop normally in ordinary environments. This lays the foundation for further breeding of FOXN1 immunodeficient rabbits in the future.
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Characterisation of the immune microenvironment and immunotherapy for cholangiocarcinoma: mechanisms, challenges and prospects

wuxueliang

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Cholangiocarcinoma is a malignant tumour with biliary epithelial features. Currently, early diagnosis of cholangiocarcinoma is difficult and the outcome of treatment is poor. Its microenvironment includes abundant fibrotic mesenchyme and a variety of cell types, and these components promote the development and metastasis of cholangiocarcinoma by interacting with tumour cells through mechanisms such as facilitating migration, suppressing immune response and inducing angiogenesis and lymphangiogenesis. Immunotherapy is one of the important means of current tumour treatment, and immunotherapy for cholangiocarcinoma has made some progress. This article reviews the characteristics of the immune microenvironment of cholangiocarcinoma, its relationship with immunotherapy and cutting-edge therapeutic strategies.
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RANBP9 targeted the expression of TGF-β1 induced the cell apoptosis in colorectal cancer Colo320 Cell

wuxueliang

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objective: To investigate Ran binding protein in microtubule-organization center (RANBP9) targeted the expression of transforming growth factor β1 (TGF-β1) and its effect on colorectal cancer Colo320 cell apoptosis. Methods: The expression data of RANBP9 (gene)? in 625 cases of colon cancer tissues and 20 cases of normal colon tissues in the TCGA database (TCGA) were analyzed. KMPLOT was used to analyze the relationship between the expression of RANBP9 and the survival time of colon cancer patients. The human protein immunohistochemistry database was used to analyze the expression of TGF-β1 in normal colon tissues and colon cancer tissues. The UALCAN database was used to analyze the relationship between the expression of TGF-β1 and the survival of colon cancer patients.The dual luciferase experiment analyzes the relationship between RANBP9 and TGF-β1.The pcDNA3.1-GFP-RANBP9 and pcDNA3.1-GFP-RANBP9-NC were transfected into the cells in the experimental group and the control group,respectively. And the normal group of cells will be established without transfection operation and routinely cultured.MTT method was used to detect the growth viability of each group of cells, flow cytometry was used to detect the apoptotic rate of each group of cells, JC-1 staining was used to detect the mitochondrial membrane potential of each group of cells, and Western blot was used to detect the expression of RANBP9 and TGF-β1 in each group of cells. Results: The expression of RANBP9 in colon cancer tissue was significantly reduced. Compared with patients with low RANBP9 expression, colon cancer patients with high RANBP9 expression had a higher survival curve, and the expression of TGF-β1 in colon cancer tissue was significantly increased. Compared with patients with high TGF-β1 expression, patients with low TGF-β1 expression had a higher survival curve, and the differences were statistically significant (all P<0.05).In colon cancer, RANBP9 can target the expression of TGF-β1. Compared with the normal group, the growth viability of the experimental group, the mitochondrial membrane potential, and the expression of TGF-β1 were significantly down-regulated, and the apoptosis rate and the expression of RANBP9 were significantly increased. The differences were statistically significant (all P<0.05).Conclusion: RANBP9 can target the expression of TGF-β1, promote the growth of colon cancer cell Colo320 and decrease the mitochondrial membrane potential, and induce its apoptosis.
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Hypoglycemic effect of Chinese Yam polysaccharide on DEX-induced IR-3T3-L1 fat cell

LIU Xinyue, WANG Fengzhong, ZHENG Hanwen, Alberto Carlos Pires Dias, FAN Bei, Wang Qiong

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Objective To examine the hypoglycemic effect of Chinese yam polysaccharide on 3T3-L1 insulin resistance cell model. Methods After the creation of the IR-3T3-L1 cell model stimulated by DEX (1 μmol/L). Followed the Chinese yam polysaccharide treatment, the glucose intake and the levels of TC, TG, LDL-C, HDL-C, GSH-Px, MDA, HK and PK were measured. Additionally, the AMPK-PI3K/Akt signaling pathway"s associated genes were identified using qRT-PCR. Results The glucose consumption of IR-3T3-L1 cells was considerably decreased after 48 hours of treatment with 1 μmol/L DEX compared to the control group (P < 0.01), and the reduction persisted for 60 hours. When CYPS at 0.05, 0.15, and 0.45 mg/mL was applied to IR-3T3-L1 cells, the following outcomes were observed: the significantly increased glucose consumption (P < 0.01), HK, PK, GSH-Px enzyme activities (P < 0.01), and HDL-C content (P < 0.01). There was a reduction in the MDA activity (P < 0.01) and the concentrations of TG, LDL-C, and T-CHO (P < 0.01). Additionally, CYPS administration dramatically decreased the levels of PI3K, Akt, GLUT-4, AMPK, IRS-2, PPARa, and Adiponectin mRNA expression (P < 0.05), and reduced the IRS-1, GSK-3β, ACC, and FAS mRNA expression levels (P < 0.01). Conclusions In IR-3T3-L1 cells, CYPS can reduce oxidative stress, control lipid metabolism disorders, and enhance DEX-induced glucose intake. The AMPK-PI3K/Akt signaling pathway"s associated genes may be connected to the process.
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Progress in the study of the mechanism of acupuncture in regulating ferroptosis after ischemic stroke

XUE jing, GAO ying, WANG shu

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Ischemic Stroke(IS)presents a complex pathogenesis,posing numerous challenges in treatment and rehabilitation.Acupuncture, as a traditional Chinese medicine treatment, has garnered widespread attention for its effectiveness in treating IS, particularly in regulating ferroptosis. Ferroptosis is a type of cell death dependent on iron and lipid peroxidation, closely linked to neurological impairment following IS. This article reviews the primary mechanisms of ferroptosis post-IS, emphasizing the interconnections between iron metabolism, lipid peroxidation, mitochondrial damage, and ferroptosis. It further explores the regulatory mechanisms of acupuncture on ferroptosis after IS, including its roles in enhancing mitochondrial function, modulating iron metabolism, alleviating oxidative stress, suppressing inflammatory responses, and influencing various ferroptosis signaling pathways. This aims to provide more scientific evidence and theoretical support for comprehensive therapeutic strategies for IS.
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Effects and comparison of ginsenosides Rg1 and Rb1 depression and anxieti-like behavior induced by chronic unpredictable stress in rats by regulating inflammation

Bei Xueyi, Jiang Ning, Yao Caihong, Zhang Yiwen, Sun Xinran, Luo Yanqin, Li Liang, Xie Mengzhou, Liu Xinmin

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Objective: To study the effect and comparison of ginsenosides Rg1 and Rb1 on depression-like and anxiety-like behavior in chronic unpredictable stress-induced rats by regulating inflammation. Methods: Seventy SPF grade SD male rats were tested for sugar and water preference after 5 days of adaptation. The animals were divided into 7 groups according to the sugar and water preference index, namely blank control group, model group, positive drug group, ginsenoside Rg1 30 μmol/kg dose group, ginsenoside Rg1 60 μmol/kg dose group, ginsenoside Rb1 30 μmol/kg dose group, ginsenoside Rb1 60 μmol/kg dose group. Except for the blank control group, the other rats were randomly subjected to 1-2 different stimulating factors every day for a total of 35 days. On the 36th day, behavioral experiments such as sugar and water preference, open field experiment, novel environment feeding inhibition experiment, elevated cross maze experiment and forced swimming were conducted to investigate its anti-depression and anti-anxiety effects. The serum and hippocampal levels of IL-1β, IL-6, TNF-α and serum corticosterone (CORT) were measured by Elisa. Result: Compared with model group, ginsenoside Rg1 and Rb1treatments significantly increased the sucrose consumption in the sucrose preference test and decrease in immobility in the forced swimming test., the latency to eat in novelty-suppressed feeding test of ginsenoside Rg1 (60 μmol/kg) group was significantly reduced, and percentage of open arm entries and time in elevated plus maze test of ginsenoside Rg1 (30 and 60 μmol/kg) groups was significantly increased. The content of serum corticosterone in ginsenoside Rg1 and Rb1 dose groups was significantly decreased. The levels of serum IL-1β and IL-6 in ginsenoside Rg1 (30 μmol/kg) dose group were significantly decreased, The levels of TNF-α and IL-6 in serum of ginsenoside Rb1 (30 μmol/kg) group were significantly decreased. And the contents of IL-1β, IL-6 and TNF-α in hippocampus of ginsenoside Rg1 and Rb high dose group were significantly decreased. Conclusion: Both ginsenosides can regulate the HPA axis and inhibit neuroinflammation, improving depression and anxieti-like behavior in rats induced by chronic unpredictable stress. In addition, ginsenoside Rg1 has a significantly better anti-anxiety effect than Rb1.
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Gualou Xiebai Baijiu Decoction regulating intestinal microflora and its metabolites to improve atherosclerosis in mice

chenzhifan, chenyulin, niesha, sunwenhao, lichang, mazishan, hukai, heyingying, liuying, tangyaoping

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Objective To explore the mechanism of Gualou Xiebai Baijiu Decoction in improving atherosclerosis (AS) in mice by regulating gut microbiota (GM) and its metabolites. Methous Thirty two male Apoe-/- mice were randomly divided into blank group, model group, atorvastatin(Ato) group and Gualou Xiebai Baijiu Decoction (GXB) group, with 8 mice in each group. Except for the Blank group, three groups of mice were established with AS models and administered by gavage according to their respective groups. Oil red O staining was used to detect the area of aortic plaques. HE staining was used to observe pathological changes in aortic tissue. Using 16S rRNA gene sequencing technology to analyze mouse GM. Detection of mouse GM metabolites trimethylamine oxide (TMAO), short chain fatty acids (SCFA), and serum levels of TG, TC, LDL-C, HDL-C, and NO. Results Compared with the Blank group, both the Model group and Ato group showed an increase in AS plaque area (P<0.05); The serum levels of TG, TC, and LDL-C in the Model group increased (P<0.001), while the levels of HDL-C and NO decreased (P<0.01, P<0.001). Compared with the Model group, the Ato group and GXB group showed a decrease in plaque area (P<0.05); The levels of serum TG, TC, and LDL-C decreased (P<0.001) and the levels of NO were increased (P<0.01) in the Ato and GXB groups; The HDL-C level in the GXB group was increased (P<0.05). Compared with the Ato group, the plaque area in the GXB group was decreased (P<0.05) and the NO level increased (P<0.01).There are 6345 characteristic sequences obtained from 16srRNA. α Diversity analysis suggests that GXB can reduce the richness of GM in AS mice (P<0.001) and improve its uniformity (P<0.05). β Diversity analysis suggests that the microbial community structure of the GXB group is similar to that of the Blank group. There are differences in the abundance of microbial communities among different groups at the phylum and genus levels. At the phylum level, the abundance of Proteobacteria was increased (P<0.01) in mouse AS modeling, while GXB intervention reduced its abundance (P<0.01) and increased the abundance of Verrucomimicrobiota (P<0.05).. At the genus level, GXB intervention can effectively increase the abundance level of Akkermansia (P<0.05). Compared with the Model group, the SCFA level in the GXB group significantly increased (P<0.01), while the TMAO level significantly decreased (P<0.01). Conclusions Gualou Xiebai Baijiu Decoction can regulate intestinal flora and intestinal flora metabolites SCFA, TMAO to improve AS.Akkermansia may be a key bacterial genus for GXB to improve AS through gut microbiota.
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Chronic myocardial ischaemia mouse model of coronary artery disease and evaluation methods

wang zhao bo, panyi, linqian, zhongjuying

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Objective To establish a stable mouse model of chronic myocardial ischemia, and to preliminarily elaborate the electrophysiological principle of T-wave flattening under ischemic state. Methods 1. C57BL/6 J mice were used as control group with normal diet for 3 months. APOE-/- mice were randomly divided into a model group and a Lipid-lowering Drug (LLD) group, and were fed a high-fat diet for 3 months; 2. Plaque condition was assessed by Hematoxylin-eosin staining (HE) and oil red staining; 3. Electrocardiograms of the mice before and after modelling; and 4. Recordings of the ventricular myocytes' action potentials. Results 1. Cholesterol (CHO) and low-density lipoprotein (LDL-C) were significantly elevated in the model group, and lipid plaques appeared in the aorta of the model mice after modelling, while the lesions in the LLD group were significantly reduced, and no plaques were found in the control group; 2. There was no significant change in the T/QRS wave (T/QRS) in the ECG after modelling in the control group, whereas a significant reduction in T/QRS was observed in the model and LLD groups. 3. There was a significant positive correlation between the degree of T/QRS depression and the levels of lipids and the degree of atherosclerosis (AS) pathology. 4. The inner layer of the cardiomyocyte action potential repetitions more slowly than the outer layer of cardiomyocytes, whereas the repetitive rate was accelerated by the ischemic inner layer of cardiomyocytes. cardiomyocytes accelerated the repolarisation rate. Conclusions APOE-/- mice can be used as a mouse model of chronic myocardial ischemia, and the increased repolarisation rate of the inner myocardium in ischemia may be the main reason for the depression of the electrocardiographic "T wave" in ischemia.
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Analysis and reflection on the quarantine and supervision policy of imported experimental animals in Japan

DING Ning, LIANG Jiaqi, FU Hongkun, WANG Ying, ZHU Daozhong, WANG Chunxia, REN Jiao, LIN Zhixiong, TAO Yufeng, MA Lidan, GAO Zhiqiang, WU Xiaowei, Yu haiqiong

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Japan is an important trading partner of China's import and export of experimental animals, and its quarantine supervision policy of experimental animals is meticulous and strict. In this paper, experimental dogs, cats and monkeys are taken as examples to make an in-depth analysis of the regulatory policies of major experimental animals for export to Japan. Through the current experimental animal related laws and regulations, inbound and outbound management measures, experimental animal breeding production and management situation and import and export business trend thinking, from the perfect experimental animal laws and regulations and inbound and outbound management measures, safeguard security, improve the management level of experimental animal breeding and promote experimental animal import and export trade. In order to provide reference for comprehensively improving the level of production, use and management of laboratory animals in China and strengthening the work exchange of laboratory animals between China and Japan.
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Inhibition of gallic acid on lipopolysaccharide-induced inflammatory response in human THP1 macrophages

DIAO Jiawen, XU Hui, MA Xinyue, QUAN Jishu

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Objective To investigate the inhibitory effect of gallic acid (GA) on lipopolysaccharide (LPS)-induced inflammatory responses in human THP1 macrophages. Methods THP1 monocytes were differentiated into macrophages with phorbol myristate acetate (PMA) and then the macrophage inflammation model was established with LPS induction，and GA was given in different concentrations. The safe concentrations of LPS and GA on THP1 cells were screened by CCK-8 method，and the normal group, model group (100 μg/L LPS ) and GA group（100 μg/L LPS + different concentrations of GA）were set up. ELISA was used to detect the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-1β(IL-1β) in the cell culture fluid of each group. Microplate method was used to detect LDH activity and NO content in cell cultures of each group, and fluorescence staining was used to detect ROS levels, cell damage and changes in mitochondrial transmembrane potential in each group.Western blotting assay was performed to detect the levels of cyclooxygenase-2 (COX-2), HMGB1,c-Jun amino-terminal kinase (JNK), extracellular-regulated protein kinase (ERK), P38 mitogen-activated protein kinase (P38), nuclear transcription factor-κB (NF-κB), NOD-like receptor protein 3(NLRP3), Caspase-1,IL-1β、Gasdermin D(GSDMD). Results Compared with the normal group, the secretion of IL-6, TNF-α, IL-1β and NO in cell cultures was increased in the model group (P<0.05), and the secretion of COX-2, HMGB1, p-ERK, p-JNK, and p-P38 and p-NF-κB, NLRP3, Caspase-1, IL-1? protein expression was elevated (P<0.05), the expression level of GSDMD protein activation fragment was reduced (P<0.05), ROS generation and cellular damage were significantly increased, mitochondrial transmembrane potential was significantly lower than that of the normal group, and the activity of LDH was elevated (P<0.05); in comparison with the model group, the cell culture fluid of the GA group IL-6, TNF-α, IL-1β and NO secretion were decreased (P<0.05), COX-2, HMGB1, p-ERK, p-JNK and p-P38 and p-NF-κB, NLRP3, Caspase-1, IL-1β protein expression was decreased (P<0.05), GSDMD protein activation fragment expression level was increased (P<0.05), ROS generation and cellular damage were decreased, mitochondrial transmembrane potential was gradually increased, and LDH activity was decreased (P<0.05).Conclusion GAhas an inhibitory effect on LPS-induced inflammatory response in THP1 macrophages, and its anti-inflammatory mechanism may involve MAPK and NF-κB signalingpathways.
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Sanguisorbae Radix alleviates damage in Ulcerative Colitis Mice Based on PPARG and SCL7A11/GPX4

Xiaoxi Yin, Zixuan Chen, Yi Yuan, Jingnan Ma, Jing Wang, Tianyi Lv, Miaomiao Tong, Li Li

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Objective: To investigate the mechanism of Sanguisorbae Radix (SR) in the treatment of Ulcerative Colitis (UC). Methods: The study analyzed GSE92415 from the Gene Expression Omnibus (GEO) database using differentially expressed genes analysis, weighted gene correlation network analysis (WGCNA) and FerrDb. Core genes were identified through protein-protein interaction (PPI) network and correlation analysis. The efficacy of SR in UC was evaluated in a dextran sodium sulphate (DSS)-induced colonic inflammatory mouse model by analyzing DAI, histopathology and colon length. ELISA was used to examine levels of inflammatory cytokines and lipid peroxidation. The expression levels of ZO-1 tight junction protein, PPARG, SLC7A11 and GPX4 protein were examined through Western blotting or immunofluorescence labeling. Results:S Nine differentially expressed genes associated with ferroptosis were screened and PPARG was identified as a key gene. The experimental animal results showed that SR significantly prevented colon shortening and ameliorated histological injuries of colons in DSS-treated mice. SR inhibited cytokine levels of IL-6 and TNF-α, improved the reduction of ZO-1 and levels of MDA and GSH in colon tissues of DSS mice. Meanwhile, it enhanced the activation of PPARG, SLC7A11 and GPX4, which reversed the therapeutic effect of DSS in mice with colitis. Conclusion: Iron death is closely related to UC. SR can inhibit iron death by regulating the PPARG and SCL7A11/GPX4 expression, thereby improving colon epithelial injury and dysfunction in UC mice. This provides ideas and direction for UC treatment strategies.
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Lipidomics driven biomarker discovery in chronic lung disease

Zhang Ang, Liu Xinguang, Du Yan, Xie Yang

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Chronic lung diseases (CLD) include chronic obstructive pulmonary disease, asthma, idiopathic pulmonary fibrosis, etc. Studies have shown that CLD is closely related to the disorder of lipid metabolism. Therefore, lipids, as biomarkers of CLD, may be of great value in the diagnosis, prevention and monitoring of disease treatment.This review mainly discusses four aspects of lipidomics, selection of clinical samples for lipidomics, discovery of biomarkers for CLD, and differentiation of TCM syndromes for CLD.
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Establishment of WHBE rabbit knee osteoarthritis model and intervention study of platelet-rich fibrin releasates

Huang Jun-jie, Chen Min-li, Zhu Ke-yan, Jiang Jing-ao, Zhang Jing-hui, Jiang Chao, Pan Yong-ming

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Objective WHBE rabbit KOA model was established by excision of medial collateral and partial patellar ligaments and intervention with platelet-rich fibrin release solution (PRFr). To investigate the mechanism of cartilage injury and inflammation in WHBE rabbit KOA model and the effect of PRFr intervention on KOA process. Methods Twenty-four WHBE rabbits were randomly divided into three groups: normal control (NC) group (n=6), KOA group (n=12) and PRFr group (n=6). The KOA group and PRFr group injected 0.5 mL of saline and PRFr into both joint cavities on the 7th and 14th postoperative days, respectively. At 4 and 8 weeks of modeling, knee joint grade score, X-ray imaging observation and gross score were performed. Serum levels of IL-1β, TNF-α and MMP-13 were detected by ELISA. Pathological sections were made after decalcification, and then HE staining, toluidine blue staining, safranin o-fast green staining and immunohistochemical expression of TGF-β, BMP3 and NF-κB were detected. Results Lequesne MG behavioral score, Mankin's score and Pelletier score of WHBE rabbits after the operation were significantly increased vs NC group (P<0.01); Pathological observation shows surface defects of cartilage and partial loss of chondrocytes; These results above indicated that the KOA model was established successfully. In KOA rabbits, knee joint swelling, joint pain stimulation and movement limitation were obvious, and X-ray showed high-density soft tissue shadow, indicating more joint effusion, and rough articular surface in general. After PRFr treatment, the levels of serum inflammatory factors (IL-1β, TNF-α, MMP-13) in KOA model rabbits were significantly reversed (P<0.05, P<0.01), and the surface of cartilage became smooth, and most of the chondrocytes were neatly distributed. At the same time, the expression levels of TGF-β, BMP3 and NF-κB induced by KOA were significantly decreased(P<0.01). Conclusion This study successfully established the KOA model of WHBE rabbits, and PRFr can improve the cartilage injury and inflammation of WHBE rabbits KOA model through TGF-β/BMP and NF-κB pathways.
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Potential risks of biosafety and prevention strategies in medical and pharmaceutical research laboratories

Chen Mo, Tao Jiayi, Wang Hongxu, Zhang Qingjian

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Medical and pharmaceutical research laboratory usually involves wide range of study aspects. The materials used in the laboratory are various, including animals, microorganisms and nano-particles and other kinds. Meanwhile, with the production of more and more waste, more biosafety risks would be created. Here, we outlined the potential safety risks related with gene amplification, gene recombination, pathogenic microorganism research, nanotechnology, animal experiments, genetically altered animals, and experimental waste, etc. It also proposed preventive measures for laboratory biosafety risks, which mainly associate with developing strict legal frameworks, bettering hardware infrastructure, strengthening safety awareness, and enhancing education and training programs.
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Research progress on related complications after snake bite

LI Yumei, YANG Yue, YI Jun, YAN Zhangren, WANG Wanchun, DONG Degang

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Snake bite is a common clinical emergency, which has the characteristics of acute onset, rapid changes in condition, high disability and mortality rates. In addition to the common systemic and local tissue damage, snake envenomation can also cause significant complications, including immediate and delayed effects. These complications are also the main causes of disability and even death caused by snake bite, which seriously affect the long-term prognosis and quality of patients’ life. This paper summarizes the symptoms, diagnosis and treatment of snakebite complications from the aspects of blood system, nervous system, motor system, endocrine system, reproductive system and other aspects, in order to provide references for effective and precise treatment of snake bite in clinical practice.
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Research progress of ferroptosis in the mechanism of cerebral ischemia reperfusion injury

jianghuan, BAI Wenya, SHAO Jianlin

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Ferroptosis is a newly cell death mode discovered in recent years, involving in a variety of pathophysiological processes, such as ischemia reperfusion injury, neurodegenerative diseases and tumors, etc. At present, there is a lack of effective methods to prevent and treat ischemic stroke worldwide, and ferroptosis which is involved in cerebral ischemia reperfusion injury. 50 articles were included in this paper after searching the related literature, which published in databases such as PebMed, Wanfang, CNKI and VIP in recent years. Discussing theIorn metabolism and the concept, mechanism and regulation of ferroptosis, the role of ferroptosis in the mechanism of cerebral ischemia reperfusion injury, and the methods of inhibiting ferroptosis, this paper attempts to provide reference for finding a new potential treatment for ischemic stroke from the direction of inhibiting ferroptosis.
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Research progress on MHC function and transgenic mouse models

CAO Xiangwen, LI Min, YIN Qi, HAN Xueliang, WANG Yuan, ZHAO Guangyu

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The major histocompatibility complex (MHC) is closely related to immune regulation, and not only has genetic polymorphisms, but also has species differences in MHC restriction. The construction of humanized MHC transgenic mouse models is an important strategy to break through the differences of MHC species and simulate the characteristics of human immune response. MHC transgenic mice are mainly divided into MHC I or MHC II. single transgenic mouse models and MHC I and MHC II double transgenic mouse models. At present, the strategy of knocking out H-2Kb and H-2Db or knocking out murine β2m is adopted to eliminate the competitive inhibition of HLA I molecules by endogenous H-2 class I molecules, and the construction of HLA II transgenic mouse model is to knock out the β strand of murine origin and transfer it to HLA II. class genes.With the optimization of construction strategies, MHC transgenic mouse models have been applied to epitope vaccine development, tumor treatment, and disease genetic association studies, becoming a powerful tool for preclinical trials. In this paper, we summarize the relevant data of MHC transgenic mouse models, and summarize the construction strategies of MHC transgenic mouse models and their application progress in vaccine development and disease treatment.
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Advances in the study of glucose metabolic reprogramming in the pathogenesis of Alzheimer's disease

Xu Qian, Yang Qiong, Tan Zihu

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Alzheimer's disease (AD) is a neurodegenerative disorder characterized by widespread dementia. Despite extensive research on the pathogenesis of AD over the past fifty years, the underlying mechanisms responsible for AD-related cellular damage and cognitive impairment remain elusive. Currently, multiple studies have confirmed alterations in glucose metabolism patterns within nerve cells in individuals with AD. This metabolic transition plays a crucial role in cell survival and disease progression, even occurring decades before pathological changes and cognitive dysfunction manifest. This article provides an overview of potential mechanisms through which glucose metabolism reprogramming contributes to AD development in various types of nerve cells and brain regions, as well as their interplay, aiming to establish a foundation for further investigation into AD while offering insights and ideas for the development of novel preventive and therapeutic approaches.
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Analysis of animal model of psoriatic arthritis based on the characteristics of Chinese medicine and Western medicine

Shi Yajin, LI Hanbing, LI Genlin, WU Suhui

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Purpose: In recent years, with the incidence of psoriatic arthritis gradually increasing and incurable in China, it has become a major problem in the medical career. Therefore, In this study, we actively explored the pathogenesis of psoriatic arthritis based on the characteristics of Chinese and Western medicine clinical evidence, summarized the currently available animal models and analyzed and evaluated them. Methods: Literature detected in line with this topic was collated and summarized, and the existing models were given an agreement score through the study of the etiology and pathogenesis of psoriatic arthritis in Chinese and Western medicine, the diagnostic criteria of Chinese and Western medicine, and the analysis of the characteristics of the animal models and their degree of agreement with the clinic. Results: This thesis found that the human leukocyte antigen transgenic mouse model, the multiple hybridization transgenic mouse model, and the mannan-induced mouse model had the highest fit scores. Conclusion: Since psoriatic arthritis is mostly seen in Europe, the preparation methods of animal models are mostly imported from abroad, and there are very few animal models prepared with the characteristics of Chinese medicine, therefore, the model agreement scores of western diagnosis are higher than those of Chinese medicine diagnosis as a whole. It is hoped that in the future, we can combine the unique diagnosis and treatment methods of Chinese medicine to further improve the types of animal models of psoriatic arthritis, so as to provide a basis for the construction of more ideal animal models for the treatment of psoriatic arthritis by combining traditional Chinese medicine and Western medicine.
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Research progress of mitochondrial quality control in respiratory diseases

xujingjing

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Respiratory diseases (lung inflammation, pulmonary fibrosis) are a serious threat to human health. As organelles unique to eukaryotic cells, mitochondria not only have important functions in energy production, biosynthesis and maintenance of intracellular homeostasis, but also act as diverse signaling organelles involved in inflammation, proliferation, differentiation, cell repair and other processes. The mitochondrial quality control system includes mitochondrial biogenesis, mitochondrial dynamics, and mitochondrial autophagy. It has been found that certain pathological mechanisms of respiratory diseases such as oxidative stress and inflammation are closely related to dysregulation of mitochondrial quality control. Therefore, this paper summarizes the research progress of mitochondrial quality control dysregulation in respiratory diseases (chronic obstructive pulmonary disease, pulmonary fibrosis, acute lung injury, asthma and bacterial pneumonia), in order to seek new ideas for the prevention and treatment of respiratory diseases from the perspective of mitochondria.
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Neuro-pathological study of the intrathalamic neurovirulence test of poliomyelitis vaccine in rhesus macaques

YU Pin, XU Yan-feng, HAN Yun-lin, ZHAO Wen-jie, QIN Chuan

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Objective To study the effect and pathological mechanisms of the neuro-immune response of viral encephalomyelitis caused by the virulence reversion of the intrathalamic neurovirulence test of poliomyelitis vaccine in rhesus macaques. Methods The stock solution (not less than 7000 lgCCID50?L-1) of the inactivated polio vaccine (Vero cells) of type I, type II and type III Sabin strains and 10-1 dilution of each type of polio vaccine were carried out using intrathalamic neurovirulence test. We observed the pathological changes of polio. Via immunohistochemical method, the distribution of receptor CD155 and CD4+ T lymphocytes, CD20+ B lymphocytes and CD68+ microglia were detected. Results Lesions were observed on the virulence reverted polio case. Inflammatory cells infiltrating, neuronal degeneration and necrosis, satellite phenomenon, perivascular cuffing and glial cell proliferation were observed in the spinal cord. The inflammatory cells were mainly CD4+ T lymphocytes, CD20+ B lymphocytes, and CD68+ microglia in perivascular cuffing and proliferative glial nodules. There is no significant difference on distribution of poliovirus receptor CD155 in neurons and glial cells of monkeys with and without polio, while no expression was observed in vascular endothelial cells. Conclusion Polio caused by virulence reversion of the intrathalamic neurovirulence test is viral encephalomyelitis.
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Effects and mechanism of Five-elements music on the social behavior of offspring from Stress-Injured pregnant rats

WU Yongye, YANG Liping, ZHAO Jiajia, SONG Qi, HOU Junlin, WANG Yaohui, LI Xiaolin

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Objective To investigate the effects and mechanism of Five-element music on the social behavior of Pregnant Children with Allergic Stress during Pregnancy, in order to provide a basis for the early prevention and treatment of clinical fetogenic affective disorders. Methods Forty-five pregnant mice were randomly divided into three groups: a blank group, a model group, and a Five Element Music group. In the model and Five Element Music groups, the model was induced using the bystander electric shock method. Additionally, the Five Element Music group was exposed to Palace Tune Five Element Music daily from 17:00 to 19:00 during pregnancy. On the 19th day of pregnancy, ELISA was employed to assess the levels of adrenocorticotropin (ACTH) and cortisol (CORT) in the serum of pregnant mice in each group for modeling evaluation. The offspring were subsequently grouped with their mother mice and underwent an 8-week-old three-box social experiment to observe social behavior. We utilized the immunofluorescence double-labeling method to detect glutamatergic neuron activity in the mPFC region of the offspring mice. Furthermore, high-performance liquid chromatography was employed to measure the total glutamate (Glu) content in the mPFC region of the offspring mice, while Gorky staining was used to observe changes in dendritic spines of mPFC neurons in the offspring mice.Results Compared to the blank group, the model group exhibited a significant increase in the levels of ACTH and CORT in the serum of pregnant mice, alongside a significant decrease in the social interaction time and social novelty preference index of the offspring mice. Additionally, there was a significant decrease in the activity of glutamate neurons, glutamate content, and neuronal dendritic spines.In contrast, when compared to the model group, the Wuxing Music group demonstrated a reduction in the levels of ACTH and CORT in the serum of pregnant mice, as well as improvements in the social behavior, glutamate neuron activity, glutamate content, and the condition of neuronal dendritic spines in the offspring. Conclusions The intervention of Five-element music effectively ameliorates offspring"s social behavior disorder resulting from prenatal fear stress, with its mechanism potentially linked to enhancing glutamate neuron activity in the mPFC region.
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Immune cell composition analysis of CAST/EiJ mice

LIUQI, HOU Yongzhi, LI Na, ZHANG Jingjing, LU Jiahan, CONG Zhe, ZHU Lin, XUE Jing

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Objective To explore the possible causes of susceptibility to multiple pathogens in CAST/EiJ mice, immune cell phenotypes in the peripheral blood and spleen of CAST/EiJ mice were analyzed to clarify the immune cell composition of CAST/EiJ mice. Methods Classical dendritic cells (cDCs), natural killer (NK) cells, B lymphocytes, T lymphocytes and their subsets in peripheral blood and spleen of CAST/EiJ mice and C57BL/6J mice were detected by flow cytometry using the cell surface markers CD3, CD4, CD8, CD11b, CD11c, CD19, CD27, CD49b and TCRβ. Results There was no significant difference in the proportion of cDCs between CAST/EiJ and C57BL/6J mice, but the cDC1 cell subset was low. The proportion of NK cells (mainly mature NK cell subsets) and T lymphocytes (mainly CD8+ T cells) were both lower in CAST/EiJ mice than in C57BL/6J mice, while the proportion of B cells in CAST/EiJ mice were higher than those in C57BL/6J mice. Conclusion The proportion of NK and T lymphocytes in CAST/EiJ mice was lower than those in C57BL/6J mice.
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Evaluation of an animal model of chronic fatigue syndrome based on data mining

Li Ya-qing, Wang Can, miaomingsan

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Objective To study the application of the animal model of chronic fatigue syndrome, and to provide reference for its animal experimental methods and model improvement. Methods Using "chronic fatigue syndrome, animal model" and "chronic fatigue syndrom, animal model" as search terms, we searched in China Knowledge, Wanfang and PubMed databases. Literature on animal models of chronic fatigue syndrome was compiled from July 2000 to July 2023, and experimental animals, modeling methods, positive drugs, and detection indexes were sorted out, and a database was established for statistics and analysis. Results A total of 155 articles were included; the most experimental animals were male SD rats of SPF grade and 6-8 weeks of age, with weights of 180-220g; the most used mice were male KM mice of 18-22g and 6-8 weeks of age; the most used positive drug was ginseng saponin tablets; the most used modeling method was the method of "forced cold water swimming + chronic restraint"; the most used modeling method was the method of "forced cold water swimming + chronic restraint"; the most used modeling method was the method of "forced cold water swimming + chronic restraint". +The most widely used modeling method is the "forced cold water swimming" method, and the modeling cycle is mostly concentrated in 14 days, and most of them adopt the method of intervention while modeling; the testing indexes are mainly behavioral, which mainly include the exhaustion swimming experiment, the open field experiment, the rat-tail suspension experiment, the Morris water maze experiment, and the observation of the general state of the animals, and the second is the serum biochemical indexes, and the most frequently detected indexes are serum MDA, SOD, TNF-alpha, IOD, and IBP, TNF-α, IL-1β, CORT, IL-2, IFN-γ, ACTH, GSH-Px, IgG, IgA, IL-6, etc. The rest of the indicators were selected according to the research purpose, including the indexes of spleen, thymus, etc., the morphological indicators detected by HE staining and ultrastructural electron microscopy, the immunohistochemistry-related indicators, the CRH and Smilax detected by fluorescence PCR, the CRH, the Smilax, the Smilax, and the Smilax. The mRNA expression indexes of CRH, Smad4, BDNF, CREB, IFN-γ, NRF-1, TFAM, Tlr4, etc., and the protein expression indexes of BDNF, CREB, ERK, Bmal1, Clock, NGF, PGC-1α, SIRT1, Smad4, Caspsae-3, etc., were determined by Western blot method. and other related protein expression indicators. Conclusion At present, the pathogenesis of chronic fatigue syndrome is still unclear, and the animal model is mainly based on the stress modeling method of "forced cold water swimming + chronic restraint", which simulates the physical and mental fatigue state of human beings; there is no uniform regulation on the criteria for the formation of animal models, and the evaluation of the model is based on the application of a variety of behavioral experiments individually or in combination, and the objective evaluation indexes are mostly used to argue the etiology of the disease and evaluate the effectiveness of interventions. The objective evaluation indexes are mostly used to prove the conjecture of etiology and evaluate the effectiveness of interventions; the results of the application of various test indexes show that chronic fatigue syndrome may be related to inflammatory response, neurological dysfunction, and mitochondrial dysfunction, and there may be abnormalities in the pathogenesis of immune function, energy metabolism, cell proliferation, and cell death, which is expected to provide reference for the application of the model, and to provide ideas for the model"s refinement. The excavated content is expected to provide reference for the application of the model and provide ideas for the improvement of the model.
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Application and prospect in experimental animal model of chronic stress

YUE Xiaoqi, Li Yuting, JIANG Miao, ZHAO Zigang, NIU Chunyu

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The effect and mechanism of chronic stress on psychosomatic diseases is a hot topic in the field of stress research. The establishment of animal models to simulate human chronic stress state is of great significance in the principle of stress response, the pathogenesis of related diseases, clinical treatment and drug development. In this paper, the commonly used animal models of chronic stress at home and abroad were reviewed, the classification of stressors, animal selection, model construction, pathological manifestations and evaluation indexes were summarized, and the advantages and disadvantages and application progress of various models were discussed, hoping to provide references for the study of model selection of chronic stress response.
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Technological advances in the study of post-stroke neural loop

Chenyanxi, Xuzhidong, Liutingting, Maliansu, Sunfangling, Wangwen

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Neural loops are formed by interconnections between neurons through synaptic structures, Which are the basic units of information transmission and processing in the brain and play an important role in the regulation of neural functions. After Stroke, the neural connections between the infarcted area and the peri-infarct region and the remote area are damaged, resulting in patients at risk for neurological dysfunction or even disability. However, with advances in detection technology, more and more studies have demonstrated that patients with stroke can produce some functional recovery during the chronic phase, possibly related to the re-establishment of synaptic connections and neural circuits. Therefore, the development of specific technology to identify and manipulate neuronal activity patterns, as well as the use of high temporal and spatial resolution imaging strategies to decipher these neurological processes allows us to understand the whole-brain network dynamics of stroke recovery and the mechanisms by which neural loop reestablishment occurs. Furthermore, we could neurobiologically comprehend the closed-loop relationships that underlie the development of stroke pathology to behavioral outcomes. Current technology for studying neural circuits focus on optogenetics, chemical genetics, in vivo calcium imaging, and functional magnetic resonance imaging technology. This article will introduce the working principles of these four major technologies, focus on summarizing the results of their respective applications in resolving neural remodeling after stroke, and briefly analyze the application scenarios, advantages and disadvantages, and future development trends of each technique. This paper will help clinical and basic researchers to to utilize these technologies in discovering new therapeutic strategies as well as evaluating the effectiveness of rehabilitation strategies.
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Analysis of the Role of Brain Plasticity in Improving Depression by Exercise

Chen Xianghe, LU Pengcheng, SHEN Ziming, LIU Chi, ZENG Xinyu, YIN Rongbin

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The research on the mechanism of depression is currently a focus in the field of neuroscience, degeneration of brain plasticity (decrease in volume, structural degradation, and functional disorder of the hippocampus, PFC, CG, etc.) leads to depression. Exercise is an important means of improving depression. Current research has confirmed the important mechanism of improving the volume, structure, and function of the hippocampus, PFC, CG, etc. in this process, but related research results only focus solely on changes in a certain brain volume or connectivity function, lacking a comprehensive understanding Systematic analysis of the mechanism of improving brain plasticity in exercise antidepressant effects. Therefore, this study aims to explore the role of brain plasticity in the occurrence of depression and the improvement of depression through exercise. This helps to comprehensively understand the role of brain plasticity in the occurrence of depression, and will also provide new ideas for exercise intervention in depression.
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Construction and application of patient-derived pancreatic tumor organoid model

LI Peng, HUANG Minli, TAN Dengxu, ZHANG Caiqin, ZHANG Yongbin, SHI Changhong

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Objective To construct a patient-derived pancreatic tumor organoid (PDO) and evaluate its effectiveness. Methods Collect fresh surgical specimens from pancreatic cancer patients for PDO culture; Compared the pathological and genetic characteristics of the PDO model with the primary tumors; Using the PDO model to evaluate the efficacy of clinical chemotherapy drugs and evaluate the effectiveness of the model. Results A PDO model of pancreatic cancer was successfully established. Histomorphological analysis displayed that the PDO model basically maintained the pathological characteristics of the primary tumor. Whole-exon sequencing showed that both organoids and original tumor tissue remained consistent in the gene mutation type and characteristics. Drug screening tests have exhibited that the PDO model had good sensitivity to gemcitabine and irinotecan. Conclusion A case of pancreatic cancer PDO was successfully constructed, which can reflect the histological and genetic characteristics of the original tumor, and can be used for drug sensitivity experiments in vitro, which proves the effectiveness of PDO model and is expected to imply precision medicine.
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Advances in the study of epigenetic regulatory mechanisms of astrocytes

xu liuqing, zhaopeiyuan, liuxihong, duxiaodan, fangmengyang, yuchenyang, houjunlin

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Astrocytes (AS) are the most abundant glial cells in the central nervous system and are involved in many physiology and pathology processes in the nervous system. Alterations in their phenotype are particularly important for the health of the CNS. Epigenetic mechanisms, including DNA methylation, histone modification, non-coding RNA regulation and chromatin remodeling, are closely linked to alterations in AS proliferation, differentiation, inflammation and other phenotypic features, but how these mechanisms function still needs to be explored and summarized. By reviewing the recent advances in the role of epigenetic mechanisms in AS under different physiological and pathological states, aim to provide new ideas for the understanding and treatment of related diseases.
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Experimental study of ursolic acid to ameliorate pancreatic β-cell injury in type 1 diabetic rats based on the TLR4/NF-κB pathway and Th17/Treg cell balance

songyu, zhangxiaoli, chenhuanhuan, tangcong

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[abstracts] Objective To investigate the effects of ursolic acid (UA) on TLR4/NF-κB signaling pathway and Th17/Treg cells in type 1 diabetes (T1DM) rats. Mmthods The T1DM rat model was prepared by intraperitoneal injection of streptozotocin, and randomly divided into blank (Control group) group, model group (Model group), metformin group (MET group) and UA group. General conditions such as body weight and blood glucose were recorded, and peripheral blood and pancreatic tissues were collected after six weeks of gavage to assess insulin intervention. Immunohistochemistry was used to observe the pathological changes in pancreatic tissues; horseshoe crab reagent was used to detect the changes in serum LPS content; qRT-PCR was used to detect the expression of pancreatic TLR4, MyD88, IκBα, and NF-κB p65 mRNAs, as well as the expression of the transcription factors RORγt and Foxp3 mRNAs; and Western blot was used to detect the expression of pancreatic TLR4, MyD88, IκBα , NF-κB p65 protein expression, and transcription factors RORγt, Foxp3 protein expression; flow cytometry to detect changes in the ratio of peripheral blood Th17, Treg cells; ELISA to detect changes in serum TNF-α, IL-6, IL-1β levels. Results After STZ-induced diabetic rats were treated by gavage for 6 weeks, compared with the Model group, fasting blood glucose (FBG) decreased significantly in both the MET and UA groups, and body weights increased; inflammatory infiltration of pancreatic β-cells was reduced; and the expression of TLR4, MyD88, IκBα, NF-κB p65, RORγt mRNA and protein expression was significantly decreased; LPS content was significantly decreased; IκBα, Foxp3 mRNA and protein expression was significantly increased; Th17/Treg ratio was significantly decreased; and TNF-α, IL-6, and IL-1β content was significantly decreased. Conclusion UA can improve the symptoms of rats by reducing LPS shift, inhibiting TLR4/NF-κB pathway, down-regulating RORγt and up-regulating Foxp3 expression to correct the imbalance of Th17/Treg cell ratio in T1DM rats.
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Effects of Shaoyao Gancao granule on hair growth, behavior and hypothalamic pituitary adrenal axis in mice with alopecia areata

QU Baoquan, LYU Shuying, LIN Wenjun, YANG Dingquan

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Objective To explore the effects of Shaoyao Gancao granule on hair growth, behavior and hypothalamic pituitary adrenal (HPA) axis in mice with alopecia areata (AA). Methods Forty-two C3H/HeJ mice were randomly divided into control, model, Shaoyao Gancao granule, corticotropin-releasing hormone receptor 1 (CRHR1) antagonist, and compound glycyrrhizin tablet (CGT) group. Photography, dermoscopy photography, weight weighing, behavioral measurement were taken, as well as corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH), cortisol, glucocorticoid receptor (GR), and brain derived neurotrophic factor (BDNF) were assessed. Results Compared with the model group, high-dose Shaoyao Gancao granules could improve hair regeneration and weight gain (P<0.05), increase the percentage of total exercise distance and central area exercise distance in the open field experiment (P<0.05), reduce the immobility time in the forced swimming experiment and tail suspension experiment (P<0.05), reduce peripheral blood CRH, ACTH, Cortisol levels (P<0.05), and increase the expression of GR and BDNF in the hippocampus (P<0.05). Conclusions Shaoyao Gancao granule can promote hair growth and improve behavioral performance in mice with AA. The effects may be related to downregulating the levels of CRH, ACTH, and Cortisol, upregulating the expression of GR and BDNF, and inhibiting excessive activation of the HPA axis.
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Advances in the study of CAV1 in digestive tract tumors

Wu Zhihang, Pan Haibang, Tang mingzheng, lixiaofeng, rong yao, cuiyan

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[Abstract] Digestive tract tumour is currently one of the most common types of cancer in the world, including esophageal cancer, gastric cancer, hepatocellular carcinoma, pancreatic cancer and colorectal cancer, etc. Their prognosis is not good and the treatment still needs further improvement. Caveolin-1（CAV1）has a dual regulatory effect in digestive tract tumors, which is both a tumor suppressor and a carcinogen. CAV1 plays an important role in cell proliferation, invasion, metastasis, angiogenesis and drug tolerance of digestive tract tumors, the regulation of CAV1 protein and its related signaling pathways may become one of the strategies for the treatment of digestive tract tumors. Therefore, the review analyzes the relationship between CAV1 and digestive tract tumors in terms of structure, function, expression regulation, regulation of epithelial-mesenchymal transition（EMT）and drug resistance in digestive tract tumors in order to provide new ideas for the diagnosis and treatment of clinical digestive tract tumors.
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Advances of Programmed Cell Death in Post-stroke Cognitive Impairment

sunkexin, xiaoyuqian, wanjun, chenshuying, chenlimin, wangyan, baiyanjie

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Post-stroke cognitive impairment (PSCI) is one of the common complications after stroke, which has a significant impact on the quality of life of stroke patients. However, the pathogenesis has not been fully explained. More and more evidence shows that the mechanism of programmed cell death (PCD) is related to PSCI, including apoptosis, necrotic apoptosis, pyroptosis, generalized apoptosis, PARP-1 dependent cell death and iron death. Therefore, it is crucial to have a clear understanding of various mechanisms of PCD and their relationship with PSCI, and to elucidate the role of PCD in the pathogenesis of diseases. The article reviews six PCD pathways related to PSCI, summarizes their mechanisms of action in PSCI, and elucidates the possible crosstalk between different pathways, in order to provide data basis for clinical targeting of regulatory factors in the PCD pathway for the treatment of PSCI.
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Experimental study on the health hazards of brake pad particles and their main component antimony sulfide respiratory exposure in mice

LI Yanhong, XIU Jinghui, ZHOU Li, GUO Jianguo

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Objective To study the potential harmful effects of brake source particles. Methods Grind the brake pad particles or antimony sulfide particles. Subsequently, the mice were exposed through tracheal perfusion for 1 month to observe pathological changes in in the lungs, heart, and liver, and flow cytometry was used to analyze the changes in peripheral blood macrophages and regulatory T cells (Treg). Results After exposure, there was deposition of foreign substances in the alveolar wall of mice, with obvious infiltration of inflammatory cells around blood vessels, which worsened with increasing exposure concentration. In addition, pathological changes such as vascular inflammation and microthrombosis in the heart, and hepatocyte swelling in the liver were observed. Treg cells in peripheral blood decreased and macrophages increased in the antimony sulfide group and low-dose brake pad particles group, while the proportion of M2-type macrophages decreased. Conclusions Exposure to brake pad particles and their component antimony sulfide particles has varying degrees of toxic effects on the lung, heart, and liver of mice, and has a impact on the immune system, indicating the potential health hazards of brake-derived air pollution.
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Effect of Sanjie Quban Recipe on the Keloid Model of BALB/C Nude Mice and Influence on Transforming Growth Factor-β1

Huang ChuHan, Yang DingQuan, Zhong ShiYi, Wu RuiYing, Yang ZhiShan, Fang HuiJuan, Liu QingWu

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Objective This study aimed to investigate the therapeutic efficacy of the Sanjie Quban Recipe on a keloid nude mice model and its impact on Transforming Growth Factor-β1 (TGF-β1). Methods Keloid tissue after surgical resection was transplanted into the subcutaneous back of healthy SPF BALB/C female nude mice aged 6-8 weeks, and the keloid nude mice model was established. The mice were randomly divided into 3 groups, 5 in each group. They were respectively treated with Sanjie Quban Recipe, Asiaticoside tablet and sterile pure water. After 28 days of continuous gavage, keloid tissue was exfoliated and weighed, HE staining, Masson staining, and immunohistochemical staining (TGF-β1). The difference of keloid weight between the three groups before and after treatment was compared, and the difference of collagen fiber, fibroblast number and TGF-β1 expression between the three groups after treatment was also compared. Results The weight difference of keloid in the Asiaticoside tablet group was greater than that in the control group, and the weight difference before and after keloid treatment was the largest in the Sanjie Quban Fang group compared with the Asiaticoside tablet group and the control group (P < 0.01). Compared with the control group, the collagen fibers in Sanjie Quban Fang group were loose and the number decreased. The number of fibroblasts decreased. The expression of TGF-β1 in Sanjie Quban Formula group was decreased compared with control group (P < 0.01). Conclusion Sanjie Quban Recipe has certain therapeutic effect on keloid, the mechanism may be through reducing the expression of TGF-β1 in keloid tissue, thereby reducing the proliferation of fibroblasts and the synthesis of extracellular matrix. This study provides experimental basis and theoretical basis for the clinical treatment of keloid with Chinese medicine.
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MiR-379-5p inhibits proliferation, invasion and migration of mouse breast cancer 4T1 cells

Song Yanmei, Sun Ningxin, Liu Chen, Song Yifen, Li Hongli, Yin Chonggao

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By investigating the effects of miR-379-5p on the proliferation, invasion and metastasis of mouse breast cancer 4T1 cells, we provide new therapeutic targets for clinical inhibition of breast cancer proliferation, invasion and metastasis. Methods After plasmid transfection, 4T1 cells were utilized to detect the expression of miR-379-5p using real-time fluorescence quantitative polymerase chain reaction (qRT-PCR); 5-ethynyl-2' doxyuridine (5-ethynyl-2'-deoxyuridine, EdU) cell proliferation assay and Transwell assay to detect changes in proliferation and invasion ability of 4T1 cells in each group; Migration ability of 4T1 cells after overexpression and knockdown of miR-379-5p was examined by scratch healing assay; BABL/c mice were used to establish the transplanted tumor model of breast cancer mice, and the tumor growth in vivo after miR-379-5p overexpression was observed.The transplanted tumor model of breast cancer was established in BABL/c mice, and the effects of overexpression of miR-379-5p on tumor growth and the number and size of lung metastases were observed . Results EdU results showed that knockdown of miR-379-5p enhanced the proliferation ability of cells compared with the control group, and miR-379-5p overexpression reduced the capacity of breast cancer cells to proliferate (p<0.05). Transwell and wound healing assays showed that knockdown of miR-379-5p enhanced the invasion and migration ability of breast cancer cells, and overexpression of miR-379-5p significantly inhibited the invasive and migratory ability of cells (p<0.01). In vivo tumorigenesis experiment of BABL/c mice showed that overexpression of miR-379-5p could significantly slow down the growth rate of tumor (p<0.05) and inhibit lung metastasis (p<0.01 ).In vivo experiments demonstrated that overexpression of miR-379-5p significantly slowed tumor growth (p<0.05). Conclusions miR-379-5p plays the role of tumor suppressor gene in breast cancer and inhibits the proliferation, invasion and migration of mouse breast cancer 4T1 cells.
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Study on pathogenesis and animal model building mechanism of ovarian dysfunction

yangzhihui, Hu Yang, Zong Zheng, Sun Xiangming, Song Hui, Chen Yingxiang, Xu Beilei, Zhang Wenjun, Chen Luning, Li Wenlan

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Premature ovarian insufficiency (POI), also known as "ovarian insufficiency", has a high incidence of 1%-5%, and has been on the rise in recent years, which has seriously affected women's physical and mental health and quality of life. At present, the cause and mechanism of POI are still unclear, and the methods and applications of model construction are also confusing, and most models have some shortcomings in pertinence and stability. This also greatly limits the related research and clinical diagnosis and treatment of POI. Therefore, this paper summarizes and discusses the etiology and pathogenesis of POI and the construction of POI animal models, in order to provide more reference for the study of POI etiology and pathogenesis and the selection and construction of models.
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The effect of AIM in inflammatory reaction and lipid metabolism diseases

Zhang Fan, Tian Chun yu, Wang Jingcun, La Xiaojin

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Apoptosis inhibitor of macrophage (AIM), belongs to the group B of scavenger receptor cysteine rich-super family (SRCR-SF). AIM is a soluble protein secreted by macrophages. The expression of this protein is controlled by the liver X receptor (LXR), and it plays an important role in the immune response of the body. AIM, as a secreted protein of macrophages, plays a broad role. It not only inhibits the apoptosis of macrophages, but also participates in the regulation of macrophage polarization. In addition, relevant studies have revealed that AIM is involved in various physiological and pathological processes such as inflammation, obesity, atherosclerosis and cancer; It was used as a biological diagnosis marker for diseases such as tuberculosis, liver cirrhosis and the like; Besides, it can promote lipolysis of adipose cells by inhibiting the activity of fatty acid synthase (FAS), playing an important role in the regulation of lipid homeostasis, lipid metabolism and autoimmune diseases. In this paper, we reviewed the molecular characteristics of AIM and its effects on inflammation, lipid metabolism and other related diseases, showing multiple functional characteristics of AIM to provide the basis for relevant medical research.
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Autophagy and Neurological Diseases

Liang YuYing, Huang Yong, Liu JunSheng, Ou YiLin, Li YiWen, Zhang Rui, Li Zheng, Zhang Zhi Nan

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Autophagy is the main degradation and recycling pathway of abnormal aggregates and damaged organelles in cells, which maintain the normal metabolic balance and material renewal of cells. Autophagy has neuroprotective effects and can affect the functional state of the nervous system by regulating the homeostasis, development, apoptosis and other physiological processes of neurons and glial cells. In recent years, a large number of studies have shown that nervous system diseases are closely related to abnormal autophagy, and inhibition or overactivation of autophagy affects the occurrence and development of depression, neurodegenerative diseases and schizophrenia. Understanding the mechanism of autophagy in nervous system diseases is of great significance for the prevention and treatment of related diseases. This review mainly reviews the current research progress of autophagy and the above diseases of nervous system, in order to provide reference for further research of these diseases.
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Research progress in the effects of Δ9-tetrahydrocannabinol on the blood-brain barrier

Zhang Qianyao, CHENG Hao, HUANG Yi-zhen, TENG Han-xin, ZHANG Yue, ZAHNG Rui-lin

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The psychoactive properties of cannabinoids are well known, and there has been controversy about whether cannabinoids can be used for therapeutic purposes worldwide.Δ9-tetrahydrocannabinol(tetrahidrocannabinol, THC), the main psychoactive substance in cannabis, its neurological mechanism has not been discovered until recently, and its neurological mechanism of action is still not fully understood. The blood-brain barrier(blood-brain barrier, BBB) is a very important barrier structure to protect the brain and is the first line of defense against foreign substances entering the brain. THC's lipophilic nature and its interaction with the endocannabinoid system make it more likely to act on the BBB. In this paper, we review the neurotoxic effects of THC, focusing on the effect and mechanism of THC on the BBB, and provide a theoretical basis for elucidating the neural mechanism of THC.
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Establishment and evaluation of animal model of Varicocele and Erectile dysfunction

lujie, liqunsheng, zhoulei, lidongrun, shenzhennan, lininghua, zhufanyu, chendong, tangwenping, yangwentao

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Objective By surgery inducing Varicocele(VC) in SD rats and Apomorphine Test(APO Test),we screened the rats with Erectile dysfunction(ED) after VC,and explored the method of establishing VC and ED model.Methods Randomly divide 60 rats into control group,sham group,and model group,with 20 rats in each group.Referring to the Turner method,partially ligate the left renal vein to induce left VC.And conduct three APO Tests to screen rats with ED after inducing VC,observe and record the number of erection,genital grooming,and yawning.Measuring the diameter of spermatic vein.Weighing both testises and kidneys.HE staining and observing the pathological changes in the penis and left testis.The success rate of modeling was calculated in the model group.Results 15 out of 20 rats in the model group were successfully established VC and ED model,with a success rate of 75%.After modeling,the diameter of the left spermatic vein in the model group increased significantly and were significantly higher than before-modeling,and the diameter of the right spermatic vein in the model group increased and were higher than before-modeling.The weight of the left testises in the model group significantly decreased compared to the right testises.There was no significant difference between or within groups in the comparison of bilateral kidney weight. In the model group,the number of erections,yawning and genital grooming decreased significantly,and decreased with the time of modeling.The pathological changes of the left testis and penis were significant in the model group.Conclusions Turner method can induce the increase of the diameter of spermatic vein in rats,causing testis injury and weight loss,and APO Test can screen the rats with ED after VC induction.The combination of the two methods is suitable for establishing an animal model with VC and ED status similar to human.
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Establishment and evaluation of hyperuricemia mouse model based on triple method

ni jianyu, bai ningning, liu xianli, gong lihong, shou qiyang

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Objective: To investigate whether a stable and reliable hyperuricemia model can be established in mice with ICR background by triple modeling method (combined with potassium oxazine, hypoxanthine and 30% yeast paste), and to evaluate the modeling method with positive drug febuxoesta. Methods: The hyperuricemia model of ICR mice was established by using potassium oxazine, hypoxanthine and 30% yeast paste diet, respectively, and the serum uric acid, creatinine, xanthine oxidase (XOD), uric acid oxidase (UOX) and other indicators were detected to evaluate the success of hyperuricemia model.Results: The serum uric acid level of ICR mice was not significantly changed by potassium oxazinate alone, which showed an increasing trend but no significant difference with that of 30% yeast paste diet and hypoxanthine combined groups, while the serum uric acid level in triple administration group was significantly increased at 7 days (P<0.01). After 14 days of dynamic monitoring, blood uric acid level of triple dose induced ICR mice peaked at 7 days. Meanwhile, the activity of XOD enzyme was increased, while that of UOX enzyme was decreased (P<0.001). In addition, triple-dose induced hyperuricemia in ICR mice was sensitive to the positive drug febuxoat, with a significant decrease in blood uric acid levels (P<0.001). Conclusion: The hyperuricemia model of ICR mice can be stably induced by triple administration at 7 days.
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Optimization and evaluation of an acute obstructive suppurative cholangitis model in rats

yukui, Liangxiaoqiang, hanmian, zhangjingzhe

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Objective A stable model of acute obstructive suppurative cholangitis in rats was established to detect its pathophysiological indexes and provide a stable and reliable standardized animal model for the study of acute cholangitis and cholestasis.Methods SPF grade male SD rats were selected and the model was constructed by injection of toxoid into the lower bile duct and ligation of the common bile duct, and the changes in body weight, mortality, major indexes of liver function and histopathological changes in the liver were evaluated before and after modeling.Results After modeling, the body weight of rats in the model group decreased significantly; the number of rats died was less; the main indexes of liver function and liver pathology showed obvious changes of cholestasis and liver function damage.Conclusion In this study, a rat acute obstructive suppurative cholangitis model was successfully established, which has the advantages of easy operation, low damage, low mortality and high successful modeling rate, and can provide a standardized experimental animal model for the study of the mechanism of many common diseases and drug development.
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Research progress of pyroptosis in renal ischemia-reperfusion injury

sunhaojie, zhujunlei, liwei, wangsuogang

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Pyroptosis is a programmed cell death mediated by cysteine proteases, which is characterized by mitochondrial participation, inflammasome assembly, plasma membrane perforation, and inflammatory release. As an important mechanism mediating the body's inflammatory response, pyroptosis plays a key role in renal ischemia-reperfusion injury (RIRI). This article reviews the molecular mechanism of pyroptosis, the mechanism of pyroptosis in RIRI and the research progress of therapeutic drugs in recent years, aiming to provide theoretical reference for the early treatment of RIRI.
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Research progress of mitochondria function regulation by TRP channels and its relationship with cardivascular diseases

zhumeiping, zhangshizhong

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TRP channel is a non-selective cation channe. In recent years, a large number of studies have found that TRP channels take participate in variety of cardiovascular diseases, and the study on mitochondrial function regulation by TRP channel and its relationship with cardiovascular diseases has become a research hotspot. Up to date, the relative references are mainly focused on TRPV, TRPM and TRPC channels. This review focuses on above TRP channels in the regulation of mitochondrial function and their relationship with cardiovascular diseases.
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Exploration of mouse model construction of breast cancer complicated with depression induced by using method of 4T1 breast cancer cell line inoculation combined with chronic restraint stress

liu yan, pengmengwei, Liu gaoyuan, Yang Tiezhu, Zhang wenxian, Wuyaosong, Chen Yulong

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【Abstract】Objective To study the core behavioral symptoms, biological indicators and pathological changes of the mouse model construction of breast cancer complicated with depression induced by using method of 4T1 breast cancer cell line inoculation combined with chronic restraint stress (CRS). Methods BABL/c mice were randomly divided into control group, stress group, tumor group and stress combined with tumor group (S T). 4T1 cell lines were inoculated into the underarm of mice in tumor and S T groups. After tumor formation, mice in the stress and S T groups were subjected to CRS for 21 days. Body weight and food intake of each group were monitored during modeling. After the experiment, depression-like behavior of mice in each group was evaluated by sucrose preference test, open field test, elevated plus-maze test, and forced swimming test. After the mice were decapitated, the weight and volume of mouse tumor were measured. Serum tumor markers including carbohydrate antigen (CA199), carcinoembryonic antigen (CEA), vascular endothelial growth factor (VEGF) and related neurotransmitters including 5-hydroxytryptamine (5-HT), norepinephrine (NE) and corticosterone (CORT) were determined by using Elisa method. HE staining was used to observe the histopathological changes of hippocampus and tumor of mice. Results In mice of S T group, the body weight and food intake were significantly decreased, tumor weight and volume were significantly increased, serum tumor markers (CA199, CEA, VEGF) levels were significantly increased, pleasure and desire to explore the new environment were weakened, stress and despair behaviors were significantly increased, serum neurotransmitters 5-HT and NE levels were significantly decreased, and CORT levels were significantly increased. In addition, the cell arrangement of tumor tissue was loose, the interstitial was reduced, the pathological nuclear classification phase was increased, the arrangement and morphology of neurons in the CA3 region of hippocampus were disordered, and the nuclear vacuolation-like changes were obvious. Conclusion The mouse model of breast cancer complicated with depression induced by 4T1 breast cancer cell line inoculation combined with chronic restraint stress showed the dual typical symptoms and biological indicators of breast cancer and depression, which can provide a good model reference for the experimental study of breast cancer complicated with depression.
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Study on the effect of Buyang Huanwu Decoction in reducing oxidative stress and protecting cerebral ischemia-reperfusion injury in rats' blood-brain barrier

maxian

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Objective To explore the mechanism of Buyang Huanwu Decoction (BYHWD) in reducing oxidative stress levels to protect the blood-brain barrier (BBB) in cerebral ischemia/reperfusion injury (CIRI) rats. Methods: Middle cerebral artery occlusion / reperfusion （MCAO/R）model was established by wire embolization method in rats, and PeriCam PSI laser speckle flow imaging system was applied to detect whether the model establishment was successful. The neurological deficits of rats were evaluated by Zea longa score; The histopathological changes in the rat brain were observed by HE staining; The degree of brain edema was detected by dry and wet weight method; The BBB permeability was detected by Evansblue (EB) staining; Ultrastructural changes of BBB were observed by transmission electron microscopy; The levels of ROS, MDA, and SOD activities, which are related to oxidative stress, were detected by kits; The expression levels of MMP-9 were detected by immunohistochemical staining and Western blot; The protein expression levels of Occludin, ZO-1, and Claudin-5 in TJP were determined by double staining of immunofluorescence and Western blot. Results:BYHWD can reduce neurological deficit scores, alleviate the brain histopathological damage, alleviate the disruption of blood-brain barrier structure, prolonged the dense region of tight junction structure, attenuated the edema of the brain in the ischemic side, and reduce the BBB permeability in MCAO / R rats. BYHWD can decrease the levels of ROS and MDA, increase the activity of SOD, decrease the expression levels of matrix metalloproteinase-9 （MMP-9）, and increase the protein expression levels of Occludin, Claudin-5, and ZO-1. Conclusions: BYHWD can increase BBB tight junction protein expression levels, reduce the permeability of the BBB, protect the ultrastructure of the BBB, and reduce brain edema, and its mechanism may be related to the antioxidant stress of BYHWD and inhibition of MMP-9 activation.
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Effects of decoction of Euphorbia fischeriana Steud. and jujuba against breast cancer of different molecular phenotypes via PI3k/Akt pathway

maliwei, YAO Hongyu, CHEN Zhe, NI Shiyu, CHEN Song, LI Jing, LIU Jicheng

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Objective To explore the effect of decoction of Euphorbia fischeriana Steud. and jujuba (DEFSJ) against estrogen receptor (ER) negative (-) and ER positive (+) of breast cancer via PI3k/Akt pathway, and to provide reference for targeted treatment of breast cancer. Methods DEFSJ extract was prepared and analysed by using UHPLC-Triple Quad. DEFSJ containing serum (CS) was prepared by a method of serumal pharmacology. Different concentrations of DEFSJ-CS were applied to (ER-) MDA-MB-453 cells and (ER ) MCF-7 of breast cancer in vitro for 48 h. Flow cytometry was used to detect the distribution of cellular cycle，DNA ladder assay was used to assess the degree of apoptosis，and Western blot was used to detect the expression of PI3k/Akt pathway-related proteins. The expressions of FoxO3a, FoxO1a and Bim mRNA were detected by a real-time quantitative polymerase chain reaction (qrt-PCR) method. Nuclear translocation of FoxO3a protein was detected by a confocal laser microscopy. Results Five batches of the DEFSJ extract were analysed by using UPLC, and the results showed that the preparation technology was feasible and the quality was controllable, ensuring the accuracy of pharmacological experiments results. DEFSJ-CS can block cells in G2/M phase (P<0.05，P<0.01). The cells treated with DEFSJ-CS emerged the typical apoptotic ladder in DNA ladder experiment. Compared with negative control group, DEFSJ-CS can decrease the proteins expression of p-PI3k，p-Akt，p-FoxO3a and p-FoxO1a (P<0.05, P<0.01) and can increase the proteins expression of Bim (P<0.05, P<0.01), and can decrease the mRNAs expression of FoxO3a and FoxO1a (P<0.05, P<0.01) and can increase the mRNA expression of Bim (P<0.05, P<0.01), and can enhance nuclear translocation of FoxO3a protein in the cells. Besides, these datas all showed that DEFSJ-CS had better effect on (ER-) MDA-MB-453 cells than (ER ) MCF-7 cells. Conclusion The regulatory effect of the DEFSJ extract on anti-breast cancer is involves the PI3k/Akt pathway, and the effect is varies with phenotypic differences.
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To explore the effect of Bufei Jianpi Formula on mitochondrial damage of skeletal muscle in COPD rats based on IRS-1/PI3K signaling axis

shentingting, lisuyun, liya, xuanyinshuang, lijingmei, Ligaofeng, hanbingyang

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Objective To explore the effects of Bufei Jianpi Formula (BJF) on mitochondrial damage of skeletal muscle in COPD rats by regulating IRS-1/PI3K signaling axis. Methods 60 SPF SD rats were randomly divided into blank group, model group (COPD stable stage group), BJF group, pioglitazone group (PIO group), BJF PIO group and aminophylline group, with 10 rats per group. The stable COPD rat model was established by the method of smoking and nasal drip (Klebsiella pneumoniae). The samples were taken from the 9th week to the end of the 20th week, and the weight of the rats was measured every week. Routine sections and HE staining were performed on lung tissue and skeletal muscle tissue respectively, and the corresponding pathological changes were observed under light microscope. Lung function of rats was observed by whole body plethysmography (WBP) at week 0, 8 and 20, including tidal volume (VT) and peak expiratory flow, respectively. PEF), 50% tidal volume expiratory flow (EF50). The mRNA expression of IRS-1, Leptin, PGC1-α and PI3K in rat skeletal muscle was detected by qPCR. The expression of PGC-1α, TFAM, IRS-1, PI3K, AKT, p-AKT and Leptin in rat skeletal muscle tissue was detected by WB technique. Results Compared with the Control group, there were a large number of inflammatory cells infiltrated in the alveolar interstitium and bronchus in the Model group, some alveolar walls were broken and fused to form air cavities, and the fibrous network was destroyed. Compared with Model group, the rupture of alveolar wall and destruction of fibroid network were improved in all groups after medication treatment, and the inflammatory cell infiltration in bronchus was reduced, especially in BJF group and Am group. Compared with the Model group, the skeletal muscle pathology of each group after medication treatment could improve the arrangement space, atrophy and fracture of muscle fibers in different degrees, and the cytoplasmic staining of muscle cells was uneven, among which the BJF group had a more significant effect. Compared with Control group, PEF, VT and EF50 in Model group were significantly decreased from week 8 (P<0.01), while PEF and EF50 in BJF, BJF PIO and Am groups were significantly increased (P<0.01). Compared with Control group, mRNA expression levels of IRS-1, PGC1α and PI3K in Model group were significantly decreased(P<0.01), while Leptin mRNA expression level was significantly increased (P<0.01). Compared with Model group, Leptin mRNA expression levels in four medication groups were significantly decreased (P<0.01), while IRS-1 mRNA expression levels in BJF, PIO and BJF PIO groups were significantly increased (P<0.01). The mRNA expression level of PGC-1α in BJF and BJF PIO groups was significantly higher than that in Model group (P<0.01). Compared with Model group, the expression level of PI3K mRNA in BJF, BJF PIO and Am groups was significantly increased (P < 0.01). Compared with Control group, the protein expression levels of PGC-1α, IRS-1 and PI3K in Model group were significantly decreased (P<0.01), while the protein expression level of Leptin was significantly increased (P<0.01). Compared with Model group, the expression level of PGC-1α protein in BJF group was significantly increased (P<0.05), and the expression level of IRS-1 protein in BJF PIO and BJF groups was significantly increased compared with Model group (P<0.05, P<0.01). The expression level of PI3K protein in Am, BJF and BJF PIO groups was significantly higher than that in Model group (P<0.01), especially in BJF PIO group. Compared with Control group, the protein expressions of TFAM and P-Akt in quadriceps femoris tissue of Model group were significantly decreased, while the protein expressions of TFAM and P-Akt were increased in all treatment groups, but there was no statistically significant difference (P>0.05).Conclusion By regulating IRS-1/PI3K signaling axis, Bufei Jiempi can improve the mitochondrial damage of skeletal muscle, increase the expression of mitochondrial coactivator PGC-1α and mitochondrial transcription factor TFAM, enhance mitochondrial biosynthesis, and reduce the pathological damage of lung and skeletal muscle tissue.
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LIMK1 promotes the progression of cervical cancer by up-regulating the ROS/Src pathway

Jia Yajing, Du Naiyi, Zhao Wei, Li Yongping, Liu Yakun

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Abstract: Objective To explore the effect of LIMK1 on the progression of cervical cancer (CC). Methods LIMK1-overexpressed human cervical cancer HeLa cells were constructed and inoculated subcutaneously in nude mice. The tumor volume was observed and the expressions of NOX2, NOX4, p-Src, p-RUNX3, RUNX3, and MMP-9 proteins in the tumor cells were detected by Western blot assay. LIMK1-overexpressed HeLa cells were cultured under 5%O2 and added with antioxidants. The protein expressions of LIMK1, NOX4, p-Src, p-RUNX3, RUNX3, and MMP-9 in the cells were detected by Western blot assay. The cell migration ability was detected by scratch assay. Transwell assay was used to detect cell migration and invasion ability. Monoclonal proliferation assay was used to detect cell proliferation ability. Results The tumor volume of nude mice inoculated with LIMK1-overexpressed HeLa cells increased significantly, and NOX2, NOX4, p-Src, p-RUNX3, and MMP-9 proteins increased, while RUNX3 protein expression decreased. In LIMK1-overexpressed HeLa cells, the protein expressions of LIMK1, NOX4, p-Src, p-RUNX3, and MMP-9 were increased, RUNX3 protein expression was decreased, while the cell migration, invasion, and proliferation ability were increased. However, after adding antioxidants, the expression levels of NOX4, p-Src, p-RUNX3, RUNX3, and MMP-9 proteins, and the ability of cell migration, invasion, and proliferation were not different from those of HeLa cells with normal expression of LIMK1. Conclusion LIMK1 promotes the progression of cervical cancer by promoting the ROS/Src pathway, thereby promoting the migration, invasion, and proliferation ability of cervical cancer cells.
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Research progress of SHP2 in digestive system tumors

WANG Peng, FAN Jianchun, JIA Juming, DIAO Qingfei, WU Xueliang, Xue Jun

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【Abstract】 Currently malignant tumors have become one of the major diseases threatening human health,with disability and mortality rates increasing year by year.Protein tyrosine phosphatase 2 (SHP2) of Src homology 2, an important member of the PTP family, is a tyrosine phosphatase with a wide range of functions,whose expression is elevated in a wide range of solid tumors,and plays a important regulatory role in invasion,metastasis,proliferation,apoptosis and drug resistance.A large number of studies have shown that SHP2 plays a very important role in the genesis and development of many solid tumors,but there is no systematic report on the role of SHP2 in digestive system tumors.Based on this paper,we reviewed the biological functions and clinical significance of SHP2 in seven different tumors of the digestive system,explored its roles and mechanisms in different cancer development stages,and summarized and looked forward to the development of SHP2 inhibitors,to further search for potential targets for effective early diagnosis and gene therapy,which is of great significance for the improvement of cancer patients' survival rate.
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Research progress on the relationship between regulatory cell death and Dilated cardiomyopathy

QiuYueqing, WangZhentao

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Dilated cardiomyopathy (DCM) is one of the common primary diseases of heart failure and arrhythmias. With the continuous deepening of research in recent years, the intrinsic molecular mechanism of regulatory cell death (RCD) has gradually become clear. Researchers have found that RCD plays a very important role in the occurrence and development of DCM. RCD mainly includes apoptosis, necrotic apoptosis, Pyroptosis, Ferroptosis, autophagy, etc. They have certain correlation and can interact and regulate each other. This article provides an overview of the current research status on the mechanisms of the five RCD modes involved in DCM, in order to provide reference for future related experiments and clinical studies.
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Research progress on the interaction between gut microbiota and microRNA in inflammatory bowel disease

KONG Binghui, BAI Longzhou, YANG Li

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Inflammatory bowel disease (IBD) is a chronic intestinal disorder characterized by an immune response to factors in the intestinal environment. Dysregulation of the intestinal flora may lead to the development of inflammation. Studies suggest that fecal transplants, probiotics, prebiotics, and dietary interventions may play a potential role in reshaping the intestinal flora and treating the disease. MicroRNAs (miRNAs) participate in physiological processes, including cell development, proliferation, and apoptosis. In addition, miRNAs are important in inflammatory processes and play a role in regulating pro- and anti-inflammatory pathways. MiRNA profiles may serve as a diagnostic tool for IBD and prognostic markers for the disease. The relationship between miRNAs and intestinal flora has not been fully elucidated, and recent studies have demonstrated their roles in regulating the intestinal flora and inducing ecological dysbiosis. In turn, the flora can regulate miRNA expression and improve intestinal homeostasis. It is important to continue exploring this relationship. Therefore, the purpose of this review is to analyze the relationship between gut microbiota and miRNAs in IBD and identify possible precision-targeted therapies for IBD.
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Study on the effects of various herbs and different proportions of herbs in Huidu Yinhua Powder on methicillin-resistant Staphylococcus aureus

LI Yufen, JIANG Shuang, SONG Wu, JIANG Tao, LIU Chang, ZHOU Haofang, TANG Yating, SU Xin

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Objective: To study the inhibitory ability of Huidu Yinhua Powder, from “Orthodox Manual of External Medicine”, on methicillin?resistant staphylococcus aureus (MRSA) as well as the inhibitory effect of virulence factor α-hemolysin (Hla) activity and biofilm formation, and at the same time, to explore the optimal ratios of Huidu Yinhua Powder, to provide experimental support for the new use of the ancient formula. Methods: The inhibitory effects of Huidu Yinhua Powder and the herbs in the formula on USA300 were analyzed by minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and disk diffusion assay (K-B method). Hemolysis assay, neutralization assay, Oligomerization assay, Western Blot (WB) to verify in which form the drug inhibits the activity of the virulence factor alpha-hemolysin (Hla). The biofilm assay was performed to evaluate the inhibitory effect of Huidu Yinhua Powder on biofilm, while finally orthogonal experiments were performed to explore the optimal ratios of Huidu Yinhua Powder. Results: Huidu Yinhua Powder inhibited the MRSA strain with a MIC₉₀ of 64 mg/mL and an MBC of 256 mg/mL, with antibacterial circle diameter of 7.50±0.50 mm. Huidu Yinhua Powder inhibits Hla activity by inhibiting Hla secretion and the minimum effective concentration (MEC) was 16 mg/mL, while the MEC of biofilm was 8 mg/mL. In Huidu Yinhua Powder, Honeysuckle and Astragalus only affected the hemolytic activity of MRSA and the formation of biofilm without inhibiting the growth of bacteria, the hemolytic activity MEC and the biofilm MEC of both of them were 32 mg/mL. Glycyrrhiza had a strong bacterial inhibitory capacity, with a MIC₉₀ of 8 mg/mL and a biofilm MEC of 1 mg/mL, without showing inhibitory hemolytic activity at subinhibitory concentrations. The final orthogonal experiment showed that when the ratio of honeysuckle: astragalus: glycyrrhiza in Huidu yinhua Powder was 1:2:4, the MIC₉₀ was 16 mg/mL, the hemolytic activity MEC was 8 mg/mL, and the biofilm MEC was 4 mg/mL, all of which were the lowest among the nine groups. Conclusion: Huidu Yinhua Powder affects the hemolytic activity and biofilm formation of MRSA at sub inhibitory concentrations, with the optimal ratio of honeysuckle, astragalus, and glycyrrhiza being 1:2:4.
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Effect of vitamin D on the structure and diversity of intestinal flora in spontaneously diabetic rats

chen lin, tan hong xing, hao li ping, li yan yan

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[Abstract] Objective To investigate the effect of vitamin D (VD) on intestinal flora in spontaneously diabetic rats. Methods Zucker diabetic fatty rats (ZDF rats) were randomly divided into control group (Con), VD control group (VD), model group (T2DM) and VD intervention group (VD+T2DM). Fasting blood glucose profile and oral glucose tolerance level were detected in rats in each group. The 16S rDNA sequencing technology was used to detect the changes in rat intestinal flora, and OTU analysis (Venn diagram), Alpha diversity analysis (chao1, observed species, PD whole tree, shannon and simpson), Beta diversity analysis [principal coordinate analysis (PCoA)], flora structure and colony species variability analysis [linear discriminant analysis and influence factor (LEfSe) analysis]. Results VD intervention significantly improved fasting blood glucose levels and insulin resistance in T2DM rats (P<0.05).Alpha diversity results showed no significant differences in chao1, observed species, PD whole tree, shannon and simpson indices between T2DM and VD+T2DM groups (P>0.05) ; Beta diversity analysis showed that the VD+T2DM group had more species similarity with the Con group compared with the T2DM group. The dominant bacteria of rat intestinal flora in each group were significantly different; compared with the T2DM group, the VD+T2DM group showed decreased abundance of Bacteroidetes and increased abundance of Firmicutes and Clostridium XIVa. Conclusion VD improved fasting glucose elevation and insulin resistance in T2DM rats; VD improved the structure of intestinal flora, decreased Bacteroidetes, and elevated Firmicutes and Clostridium XIVa abundance in T2DM rats.
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Overview of in vitro skin models of transdermal drug delivery system

LiuYan, HuXiaolei, XuKeHong, ZhaoHaiRong, WuXiuMei, YangZiZhong, ZhangChengGui, ZhaoYu, GaoPengFei

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Skin modeling of transdermal drug delivery systems refers to the experimental models which can mimic the structure and function of human skin to explore and evaluate the absorption, penetration and efficacy of medicines in transdermal drug delivery. It provides alternative to traditional human skin experiments and reducing the utility of human skin in medicines research, which is convenient, controllable, and cost-effective . For the skin models of transdermal drug delivery systems, this paper introduces commonly used animal skin models, artificial skin models, and recombinant human skin models from the perspective of the transdermal absorption pathway of medicines, and analyzes its advantages and disadvantages as well as its applications, so as to provide references for the research and development of transdermal formulations and topical therapies and so on.
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AMPK: Diverse Regulatory Mechanisms and New Perspectives for Disease Treatment

Ning Jinghua, zhang xin, zhang Yuzhe

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The AMP-activated protein kinase (AMPK) is a conserved cellular energy Adenosine monophosphate, plays an important role in regulating cell growth, proliferation, differentiation, autophagy, phosphorylation, crosstalk, and glucose and lipid metabolism. AMPK is activated when the body is low in energy or under certain extreme conditions, and it's suppressed when there's an excess of nutrients to maintain the energy balance. In addition, the regulatory mechanism of AMPK signaling pathway mediating ferroptosis also reflects its unique role. AMPK plays a special regulatory function in different organelles, which provides a new direction for disease therapy. It is also a therapeutic target for the prevention of diseases such as reproductive system diseases, aging, cancer, inflammation and cardiac dysfunction. This article reviews the damage of cellular energy imbalance to the body's microenvironment. AMPK stimulates its potential therapeutic potential in diseases and drugs through diverse signal regulatory mechanisms, it provides a new way of treatment for different system diseases. This review summarizes the diverse regulatory mechanisms of AMPK signaling pathway, and provides theoretical reference for cancer therapy and other diseases therapy targeting AMPK.
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Discussion on the Mechanism of "Brain-Spleen Inflammation Coupling" in Rat Model of Acute Ischemic Stroke Stasis Toxin Syndrome

dongyilei, Liu Yue, Li Junyuan, Fu Jianhua, Zhang Yunling, Yao Mingjiang

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【Abstract】Objective: To investigate the correlation between brain injury and spleen damage in rat models of acute ischemic stroke and stasis interaction, and its effect on the signal axis of MCP-1/CCR2 chemokine, and to provide experimental basis for the mechanism of "brain-spleen inflammatory coupling" in spleen lesions caused by acute ischemic stroke. Methods: Forty male SD rats were randomly divided into sham group, carrageenan/yeast stasis syndrome group (Carrageenan/Yeast, CA/Y), middle cerebral artery occlusion group (MCAO), middle cerebral artery occlusion (MCAO), middle cerebral artery stasis syndrome group (MCAO CA/Y), 10 rats in each group. The CA/Y group and the MCAO CA/Y group were injected with 10 mg穔g-1 carrageenan, 10 mg穔g-1 intraperitoneally on the first day of molding, and 2 mg穔g-1 of dry yeast suspension were injected subcutaneously on the second day, and the MCAO group and MCAO CA/Y group were established by wire embolism on the second day. 24 h after cerebral infarction model, the neurological deficit score was performed on each group of rats, the percentage of cerebral infarction area was observed by TTC staining, the spleen weight was determined, and the correlation between the percentage of cerebral infarction area and spleen weight was further analyzed by Spearman correlation coefficient, the pathological morphology of brain tissue and spleen tissue was observed by hematoxylin-eosin (HE) staining, and monocyte-1 (monocyte) was detected in rat plasma by enzyme-linked immunosorbent assay (ELISA). chemotactic protein 1, MCP-1), interferon-γ γ (IFN-γ) content, western blot method to detect ischemic side brain tissue chemokine C-C-motif receptor 2 (CCR2) protein expression. Results: Compared with the ham group, the neurological deficit score, cerebral infarction area, MCP-1 and IFN-γ content in plasma were significantly increased (P<0.01), spleen weight decreased significantly (P<0.01), and the expression of CCR2 protein in brain tissue was also significantly up-regulated (P<0.05) in the MCAO group and the MCAO CA/Y group. The area of cerebral infarction increased significantly (P<0.01), the weight of the spleen decreased significantly (P<0.01), and the expression of CCR2 protein in brain tissue and spleen tissue was also significantly up-regulated (P<0.05), compared with the MCAO group, the area of cerebral infarction in the MCAO CA/Y group was significantly increased (P<0.01) and the weight of the spleen decreased significantly (P<0.05). Spearman correlation analysis showed that spleen weight was negatively correlated with the percentage of cerebral infarction area (P<0.01, r=-0.9711). The pathological morphological observation results showed that the pathological changes in the MCAO CA/Y group were the most serious, cerebral liquefaction necrosis foci could be seen in the brain tissue cortex, the arrangement of neuronal cells in the lesions was sparse, disordered, volume atrophy, a small number of vacuoles and nuclear solidification, most of the neuronal cells were red degeneration and necrosis, microglia hyperplasia was obvious, small blood vessels were significantly increased, and interstitial lipid degeneration was superb; The density of periarterial lymph sheath cells in part of the spleen tissue is reduced, and the marginal area is widened. Conclusion: A correlation between brain and spleen injury could be found after acute ischemic stroke with stasis and toxins syndrome, and the chemokine signaling axis of MCP-1/CCR2 might be involved in the mechanism of "brain-spleen inflammation coupling".
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Effect of lidocaine regulating Hippo-YAP signal pathway on ischemia-reperfusion injury in orthotopic liver transplantation rats

Liu Yue, Akbar Nurmaimaiti, Ye Jian-rong

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Objective: To explore the effect of lidocaine (LID) on ischemia-reperfusion injury in orthotopic liver transplantation (OLT) rats and to analyze its mechanism of action. Methods: 60 rats were randomly divided into control group, model group, low-dose LID group, medium-dose LID group, high-dose LID group, and Verteporfin group, with 10 rats in each group, except for the control group, and other rats were used to construct OLT models. The pathological changes in liver tissue was observed by hematoxylin-eosin (HE) staining, the serum aspartate transaminase (AST), alanine transaminase (ALT), total bilirubin (TBIL) and lactate dehydrogenase (LDH) activities were detected by automatic biochemical analyzer, the liver tissue inflammatory factor tumor necrosis factor-α (TNF-α), interleukin (IL) -6, IL-1β and IL-10 levels were detected by enzyme-linked immunosorbent assay (ELISA), the reactive oxygen species (ROS) was detected by fluorescence probe method, the malondialdehyde (MDA) was detected by thiobarbituric acid colorimetric method, the superoxide dismutase (SOD) was detected by nitrogen blue tetrazole colorimetry, the glutathione peroxidase (GSH-Px) was detected by spectrophotometer method, the apoptosis of liver histiocyte was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) , and the expression of mammalian STE20 like protein kinase (MST1), phosphorylation (p) - MST1, large tumor suppressor factor 1 (LATS1), p-LATS1, Yes associated protein (YAP), p-YAP, as well as apoptosis related proteins B-cell lymphoma 2 (Bcl-2) and Bcl-2 related X protein (Bax) were detected by western blotting (WB). Results: Compared with control group, the liver tissue in model group rats showed injury, liver cells necrosis and a large number of inflammatory cells infiltration, the cell apoptosis rate, serum AST, ALT, TBIL, LDH activity, liver tissue TNF-α, IL-6, IL-1β, MDA, ROS, and Bax levels significantly increased, the liver tissue IL-10, SOD, GSH-Px, Bcl-2, p-MST1/MST1, p-LATS1/LATS1 and p-YAP/YAP proteins expression levels were significantly reduced (P<0.05). Compared with model group, the liver tissue injury was reduced in low-dose LID group, medium-dose LID group, and high-dose LID group, the cell apoptosis rate, serum AST, ALT, TBIL, LDH activity, liver tissue TNF-α, IL-6, IL-1β, MDA, ROS and Bax levels were significantly reduced, the liver tissue IL-10, SOD, GSH-Px, Bcl-2, p-MST1/MST1, p-LATS1/LATS1, and p-YAP/YAP proteins expression levels were significantly increased (P<0.05). Hippo-YAP signaling pathway inhibitor Verteporfin reversed the improvement effect of LID on ischemia-reperfusion injury in OLT rats (P<0.05). Conclusion: LID may activate the Hippo-YAP pathway, which reduce inflammatory response, oxidative stress, and liver cell apoptosis, and improve liver ischemia-reperfusion injury in OLT rats.
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Overview of the research of Q fever in animal models

li jinchun, yin jiaxiang

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Q fever is a zoonotic disease caused by Coxiella burnetii infection, which is widely spread in nature. Animal models are important tools to study the etiology and pathogenesis of infectious diseases and evaluate the effectiveness of vaccines. In recent years, a variety of animal models such as invertebrates, rodents and non-human primates have been used to study Q fever. This paper summarizes the research status of different Q fever animal models, discusses the advantages and disadvantages of these models, and summarizes the requirements and standards of future modeling work.
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Advances in the application of laser speckle contrast imaging in animal models of chronic cerebral hypoperfusion

Tangxin, tanjie

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Laser speckle contrast imaging technology can be used to dynamically measure microcirculation blood flow in tissues or organs in a visual and quantifiable way. Because of its advantages of in vivo detection, real-time imaging and simple operation, it has been applied in many fields of microcirculation detection.The animal model of chronic cerebral hypoperfusion is mainly constructed by reducing the bilateral common carotid artery blood flow to the brain, which can cause long-term insufficient cerebral blood supply. Laser speckle contrast imaging technology can observe and analyze the cortical cerebral blood flow, collateral circulation opening, neurovascular coupling response, etc., in the animal model of chronic cerebral hypoperfusion , so as to judge the success of animal model modeling and the therapeutic effect of intervention methods. In this paper, the application and advantages and disadvantages of laser speckle contrast imaging technology in the animal model of chronic cerebral hypoperfusion were summarized in order to provide new ideas for clinical treatment and scientific research.
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Research progress of berberine in the field of neuroprotection in ischemic stroke

lihongyu, lanrui

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Berberine is a natural Isoquinoline alkaloid, which can be initially used as a broad-spectrum antibacterial agent in clinical treatment of enteritis, peptic ulcer, chronic gastritis, pneumonia and other diseases. In recent years, with the in-depth study of the pharmacological effects of Berberine, more and more evidence shows that Berberine has neuroprotective effects on ischemic stroke. In this review, we introduced the intervention of Berberine on risk factors of ischemic stroke, and discussed the neuroprotective effects of Berberine in different mechanisms of ischemic stroke in detail, aiming to provide some reference for clinical and basic research in this field.
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Advances in Research on Mechanisms Related to Myocardial Regeneration in Neonatal Rats

chenmengqi

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Cardiovascular disease is a health hazard to humans and systolic heart failure due to myocardial infarction is a major cause of death. It was previously thought that myocardial cells of adult mammalian heart possess a limited ability to proliferate and renew themselves, and that after ischemic injury, a large number of myocardial cells are lost and eventually replaced by non-contractile scar tissue. In contrast, it has been widely reported that mammals have the ability to regenerate myocardium which is restricted to early postnatal life, and that it is strong enough to repair damaged heart tissue. The discovery of myocardial regeneration in neonatal heart has provided an ideal animal model to investigate the mechanisms that affect myocardial cell proliferation, and subsequently many mechanisms that can reverse myocardial cell cycle arrest and promote myocardial cell proliferation have been revealed. In this paper, we review the factors affecting myocardial regeneration gene expression (ncRNAs, transcription factors, etc.), myocardial regeneration-related signaling pathways, and the regulation of myocardial regeneration by non-myocardial cells (extracellular matrix, immune response, epicardium, etc.), so as to provide directions for achieving myocardial regeneration after myocardial injury in adult mammals.
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Establishment of genetic monitoring methods for experimental quail

heyang, zhangxiulin, zhangqiuyu, zhangxiaolu, fubo, wangwendong, duxiaoyan, chenzhenwen, lichanglong

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Objective To establish a genetic monitoring method for laboratory quails. Methods Quail microsatellite loci were screened through literature search, and microsatellite DNA loci suitable for quail were screened by interspecific transfer method in quail closely related species, chickens and ducks. Quail liver tissue DNA was extracted as a template, and the corresponding loci were screened by PCR amplification and agarose gel electrophoresis. Finally, based on the amplification of the selected microsatellite loci, the number of alleles, and polymorphism, microsatellite loci combinations for quail genetic quality detection were selected and detection methods were established. Results We preliminary determined 23 microsatellite loci for genetic monitoring of closed-colony laboratory quails. Conclusion A genetic monitoring method for laboratory quails is preliminary established.
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Innovation and exploration of Medical Laboratory Animal Science teaching based on smart teaching environment

YangZiHao

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Objective In the traditional laboratory zoology lecture environment, there is less teacher-student interaction, less student interest and less engagement in learning.In order to further improve the teaching quality of laboratory zoology, Methods this teaching and research department based on the different teaching environment of multimedia classroom and intelligent classroom, the theoretical course teaching of Medical Laboratory Zoology as the research ,object, the course lecture format, teaching mode, teaching methods and other aspects of innovation and exploration. The study used questionnaires to understand changes in student engagement in learning and preferences for smart classroom use, and NVivo qualitative analysis software to code student classroom behaviour. Results The smart teaching environment resulted in higher student interest and more frequent teacher-student interaction in the classroom. At the same time, students are significantly more engaged in learning than in traditional teaching, with higher correct rates on in-class and post-lesson exercises and a better grasp of concepts related to laboratory zoology. Conclusions In summary, the smart teaching environment can bring students a better feeling and experience, improve their interest in laboratory zoology, increase classroom learning engagement and achieve good teaching results.
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Progression of tripterygium wilfordii polyglycoside in mice with premature ovarian insuffici-ency model at different time points

马林纳, makun, fanxiaodi, luojie, lijiani, zhanghan

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Objective The changes of relevant indexes in the mice model of early-onset ovarian insufficiency caused by tripterygium wilfordii polyglycoside were discussed, and the best time point for intervention was determined. Methods Forty female ICR mice were randomly divided into blank group, A, B, C, and D model groups, with 8 mice in each group. The blank group was given gavage purified water for 14d (0.01ml/10g), and the remaining groups were given tripterygium wilfordii polyglycoside suspension (80mg/kg, 0.01ml/10g) for 1d (A model group), 3d (B model group), 7d (C model group), 14d (D model group), and the materials were taken in batches. Weigh the body weight, wet weight uterus and bilateral ovaries of mice in each group; Serum FSH, LH, E2, P, AMH, INH-B and T contents were detected by enzyme-linked immunoassay. HE staining observed the number and development status of follicles and corpus luteum at all levels of mice in each group. TUNEL fluorescence staining method detected the apoptosis area in the ovaries of mice in each group. The IHC method detected the positive expression of VEGFA, CD34 and EPO proteins in the ovaries of mice in each group. The mRNA expression of HIF-1α, SDF-1 and CXCR4 in each group of mice was detected by PCR. Results Compared with the blank group, the changes of indicators in model A did not meet the POI modeling standard. The ovarian index, uterine index and body weight of mice in the B model group decreased significantly (P<0.01), the weight of the C model group decreased significantly (P<0.01), and the ovarian index of the D model group decreased significantly (P<0.05). The serum indexes of the B, C and D model groups increased the contents of FSH and LH (P<0.01 or P<0.05), and the contents of E2, PROG, AMH, INH-B and T decreased (P<0.01). The number of basal follicles, presinus follicles, sinusoidal follicles, antral follicles, preovulatory follicles and corpus luteum decreased significantly (P<0.01 or P<0.05) and the number of atresia follicles increased significantly (P<0.01) in the B, C and D model groups. The apoptosis area of TUNEL fluorescent staining in A, B, C, and D model groups increased significantly (P<0.01 or P<0.05). The positive expression of CD34, VEGFA and EPO in the B, C and D model groups decreased significantly (P<0.01 or P<0.05). The expression of mRNA in the A and B model groups of HIF-1α, SDF-1 and CXCR4 was significantly increased (P<0.01 or P<0.05). Compared with the B model group, the relevant indexes of the C and D model groups changed significantly, indicating that the C and D models were more serious and tended to develop POF. Conclusion The B model group is the turning point of ovarian function from impaired POI to irreversible POF, suggesting that 3d administration of tripterygium wilfordii polyglycoside is the best time to induce POI disease model and effective drug intervention.
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Research progress of noncoding RNA in methamphetamine-induced addiction and neurotoxicity

zhang shu wei, cheng hao, wang hao wei, miao lin, li yi, guan li na, zeng xiao feng

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Methamphetamine (METH) is highly addictive and neurotoxic, which can cause cognitive and memory dysfunction in abusers. The harm of METH lies not only in its own toxicity, but also in the high physical and mental dependence of drug addicts, often causing mental disorders and causing violent behavior, bringing great safety risks to society. Non-coding RNA (ncRNA) does not code for proteins and is an important factor in regulating gene expression at the post-transcriptional level. Studies have shown that ncRNA plays an important regulatory role in methamphetamine-induced addiction and neurotoxicity, but the specific mechanism is unclear. This article reviews the current research progress of ncRNA in regulating METH-induced addiction and neurotoxicity, in order to provide a reference for ncRNA as a forensic identification index and potential drug intervention target for METH abusers.
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Advances in the mechanism of action of cyclic RNA in Parkinson's disease

liujuan, Li Yanjie, Qin hewei, Ma luyao, Zhao nannan, Xu zhenhua

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Parkinson's disease (PD) is a neurodegenerative disease characterized by the degenerative death of dopaminergic neurons in the substantia nigra, exhibiting a range of motor and non-motor symptoms that have a serious impact on the quality of life of patients. Circular RNA (circRNA, CircRNA) is a covalent closed-loop noncoding RNA that plays an important role in the progression of PD disease. This article reviews the involvement of circRNA in oxidative stress, regulation of transcriptional levels, neuroinflammation, autophagy, and α-synuclein.
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Analysis of application characteristics of acute liver injury animal model based on data mining

huyilong, zhangshuangli, qiuguangnan, fengyifan, miaomingsan, miaojinxin

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Objective To investigate the modeling elements of various types of animal models for acute liver injury, and to provide references and suggestions for the establishment and evaluation of animal models for ALI. Methods The animal experimental literature of acute liver injury from 2002 to 2022 was searched in the databases of China Knowledge Network, WanFang, Chongqing Vip, Chinese Medical Journal Full Text Data, PubMed, etc. The animal species, positive control drugs, modeling methods, modeling drugs and drug administration of the animal models of acute liver injury involved in the literature were summarized and summarized. The results were analyzed using Excel, SPSS Modeler18.0 and Cytoscape3.8.2. Results A total of 896 papers were included in the database. The most used animal models for acute liver injury were male KM mice; the modeling methods were mainly chemical liver injury, alcoholic liver injury, drug-related liver injury and immune liver injury, and the corresponding main modeling methods were : ① intraperitoneal injection of 10.0 mL 0.1% CCl4 vegetable oil, 24 h before the experiment; ② gavage of 12.0 mL 50.0%-56.0% ethanol, 16 h before the experiment; ③intraperitoneal injection of APAP 300 mg/kg, 24 h before the experiment; ④ tail vein injection of Con A 20 mg/kg, 8 h before the experiment. The evaluation of the model was based on liver pathological indexes as the gold index, combined with biochemical indexes Serum ALT, AST, liver tissue homogenate SOD, MDA content and activity were used as direct indicators. Conclusions Since the causes of acute liver injury vary in clinical practice, the preparation of animal models of acute liver injury should be based on the specific study content and characteristics of the study, and the corresponding modeling methods should be selected.
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The mechanism of Yantiao fang regulating Rho/ROCK signaling pathway on apoptosis of intestinal epithelial cells in mice with acute gastrointestinal injury caused by sepsis

Chen Qian, Wang D, Zhang DY, Jia SL, Wang LH, Cao YJ, Li YH

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Objective To investigate the effect of Yantiao Formula on apoptosis of intestinal epithelial cells in mice with acute gastrointestinal injury caused by sepsis by regulating Rho/ROCK signaling pathway. Methods 70 BALB/c mice were randomly divided into blank group (10 mice), sham operation group (10 mice) and model group (50 mice).Mice in blank group were fed normally without treatment.Sham operation group mice only open free cecum without perforation ligation;A mouse model of acute gastrointestinal injury caused by sepsis was established by cecal ligation and perforation (CLP). The mice were randomly divided into model group, Yantiao formula low-dose, medium-dose and high-dose groups, and ROCK inhibitor group.Hematoxylin-eosin (HE) staining was used to observe the histopathological changes of mice ileum.Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) were determined by ELISA.The expression of PCNA and Ki-67 in mouse ileal cells was detected by immunohistochemistry.The expressions of Caspase 3 and Bax were detected by western blotting.The expression of ROCK mRNA and MLC mRNA in the Rho/ROCK pathway of mouse ileum were detected by RT-qPCR. Results The intestinal mucosa of the ileum tissue of mice with acute gastrointestinal injury caused by sepsis was atrophied obviously, villi were disordered, rupture and shedding were observed, epithelial cells were necrotic and shedding, inflammatory cell infiltration was obvious, and Chiu's pathological score was increased.Compared with blank group and sham group, the serum levels of pro-inflammatory mediators (IL-1β, IL-6, TNF-α) and mRNA levels of Caspase 3, Bax, ROCK and MLC in peripheral blood of mice in model group were increased, while the levels of anti-inflammatory mediators (IL-10) and expressions of PCNA and Ki-67 in ileum were decreased.Compared with model group, the histopathological changes of ileum in all Yantiao fang groups were improved to varying degrees with the increase of dose: the pathological score of Chiu's was decreased, the levels of IL-1β, IL-6 and TNF-α in serum were decreased, and the levels of IL-10 were increased.In addition, the expressions of PCNA and Ki-67 were increased, while the expressions of Caspase 3, Bax, ROCK mRNA and MLC mRNA were decreased. Conclusion Yantiao fang can inhibit the apoptosis of intestinal epithelial cells in mice with acute gastrointestinal injury caused by sepsis by regulating Rho/ROCK signaling pathway, so as to reduce the inflammatory reaction of intestinal tissue and ultimately prevent the damage of intestinal mucosal tissue.
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Research status and trend of animal models of depression: a bibliometric analysis based on CiteSpace

YANG JIAHUI, LIN MENG, LUO XIAOQUAN, LUO TAO, GONG MEIFU

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【Abstract】Objective With the help of literature metrology, the scientific literature in the field of animal models of depression was analyzed, so as to better understand the development trends and hot spots in this field.Methods We obtained publications on depression and animal models from 2013 to 2022 from the Web of Science core set (WOSCC) database. Citespace 6.1R 1(64-bit) Basic was used to analyze the annual publications, countries, institutions, authors and keywords related to this field.Results A total of 1000 articles were included in this study. From 2013 to 2022, the number of papers published gradually increased and stabilized. In terms of the number of articles, the United States has the most published articles (256) and the most Polish Academy of Sciences (50) . Wegener Gregers is considered the most influential author in the field, with 23 published articles. When analyzing keywords, “Animal Model” was the most frequent, followed by“Major depression”. Conclusions This study highlights the important research trends and hot spots in the field of animal models of depression. The US, China and Japan are leading the way in terms of publication, indicating their significant contribution to the development of animal models of depression. However, international cooperation is limited and there is more cooperation within institutions and groups. The field focuses on signaling pathways and therapeutic approaches to determine the pathogenesis of depression and better treatments. The study provides a visual analysis of trends in depression research to help researchers keep abreast of the latest developments.
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Effect of gender difference on the model of rheumatoid arthritis induced by bovine type II collagen

Sun Wenting, Wan Yingying, Yang Jiaxi, Wang Wenqian, Wang Haonan, Ye Wanting, Kou Qiuai

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Objective To construct female and male bovine collagen-induced arthritis (CIA) models and compare the effects of gender differences on the joint and extra-articular manifestations of CIA models. Methods The CIA model was induced by injection of bovine type II collagen and Freund's complete adjuvant into female and male SD rats. The general condition, arthritis index, foot swelling, serum inflammatory factors and plasminogen activator inhibitor levels, spleen index, knee and ankle joint pathology, right rear paw bone destruction, and pulmonary interstitial lesions of rats in each group were evaluated. Results The arthritis index of female CIA rats was significantly higher than that of male CIA rats on the 21st day after initial immunization (P<0.05), but there was no significant difference in the degree of foot swelling between the two groups at any time point of observation (P>0.05); Serum tumor necrosis factor α, interleukin1β and spleen index of female CIA rats were significantly higher than those of male CIA rats (P<0.001; P<0.001; P<0.05), there was no significant difference in plasminogen activator inhibitor levels (P>0.05); The scores of inflammatory cell infiltration and synovial hyperplasia in the knee and ankle pathology of female CIA rats were significantly higher than those of male CIA rats (P<0.05), and the cartilage damage of knee joint and the bone damage of right rear paw of female CIA rats were significantly higher than that of male rats (P<0.05); Both male and female CIA rats showed pulmonary interstitial inflammatory cell infiltration and mild fibrosis, but the pulmonary interstitial lesions in females were more severe than those in males. Conclusion Both female and male CIA models constructed with SD rats can have both arthritis and pulmonary interstitial lesions, but the lesion degree of female CIA rats is more serious. When using CIA models for RA related research, attention should be paid to the impact of gender differences.
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Effects and mechanism of NCTD on proliferation and apoptosis of human leukemia cells by targeting PPP5C

zhangxin, cuibingjie, yuguoxing, wangfei, zhaoliang, gaona, dujing

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Objective To study the effects and mechanism of norcantharidin (NCTD) on proliferation and apoptosis of human leukemia NB4 and K562 cells by targeting phosphoprotein phosphatase 5 catalytic (PPP5C). Methods NB4 and K562 cells were cultured in vitro, and the pcDNA3.1 and PPP5C-pcDNA3.1 plasmids were electroporated into NB4 and K562 cells. NB4 and K562 stable cell lines were screened with geneticin (G418). The protein and mRNA expression levels of PPP5C were identified by Western blot and qRT-PCR. The proliferation ability, migration ability and apoptosis rate of NB4、K562 cells were determined by CCK-8 assay, transwell assay and the Live DeadTM animal cell viability/toxicity detection kit. NB4 and K562 cells were divided into control group and different doses of NCTD group, and cultured in 1640 medium containing 0, 8, 16, 32 μg/ml NCTD. The Live DeadTM animal cell viability/toxicity detection kit detected the number of dead and live cells and cell morphology was recorded by microscope. Western blot was used to detect the protein expression levels of caspase 3, Cleaved caspase 3, JNK, p-JNK, p38, p-p38 and α-tubulin in cells of each group. Results? the proliferation ability of cells, the migration ability and apoptosis rate of NB4、K562 cells are enhanced after overexpression of PPP5C in human leukemia NB4 and K562 cells; Compared with the control group, each concentration group of NCTD promoted apoptosis in a dose-dependent manner; PPP5C overexpression antagonizes the killing effect of NCTD on leukemia cells. Conclusions NCTD can promote the apoptosis of NB4 and K562 cells and inhibit the proliferation of cells by inhibiting PPP5C.
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Research on the Mechanism of LSC Immune Escape with Intervention by TCM

ZHANG Meiling, 章, CuiYaru, ChengShupeng, LingZhiming

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Leukaemia is a critical disease with a high incidence in clinic at present, with an extremely high fatality rate. Among them, immune escape from leukaemia stemcells(LSC) is the main factor for recurrence and progression after leukaemia-induced remission. The clinical diagnosis and treatment of TCM has distinct advantages of syndrome differentiation and treatment. Based on the purpose of diagnosis and treatment, the treatment of leukaemia by TCM emphasises the "harmony of yin and yang" to estore human function, which is conducive to improving the body's autoimmunity and conforms to the mechanism of intervention for tumour cell immunity escape. This article discusses the echanism and research progress of TCM intervention in Lsc immune escape from literature research and TCM theory.
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Effects of polysaccharide content and anti-inflammatory, hemostatic and antioxidant activity before and after preparation of Mosla chinensis-jiangxiangru

Maqiuting, Xulei, Zhuyuchen, Xumengting, Zhangwenkai, Liuzhiyong

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Abstract：Objective:Exploring the changes in polysaccharides in Jiangxiangyu before and after giner juice, and evaluation of polysaccharide anti -inflammatory and antioxidant antioxidant activity before and after the processing. Methods: The content of polysaccharides beforeand after processing was determined by phenol-sulfuric acid method（Elsholtzia chinensis polysaccharide-JXRPs, Ginger Elsholtzia chinensis polysaccharide-JZJXRPs); The body internal uses the swelling model of rats and the body fat polysaccharide (LPS) to induce mousemacrophages RAW264.7 inflammation model.The best administration concentration is selected by using cell proliferation (MTT) method, Elisa method measures the expression of IL-6, IL-12, NO, IL-4, and IL-10 in the cells;Observe the bleeding time of mice to evaluate its hemostatic effect by observing the tail -cutting method; finally, use the ability to remove DPPH and ABTS to evaluate its in vitro antioxidant activity.Results: The contents of JXRPs and JZJXRPs were 13% and 22%,As a result of the swelling of rats, after 4h in injection, compared with the model group, JXRPs ppear significant difference in dosage doses of 200 mg/kg (P <0.05), and JZJXRPs When the dose is 100 mg/kg, you can significantly reduce the swelling of rats (P <0.05);The in vitro anti-inflammatory results show that the polysaccharides before and after the processing can significantly inhibit cell secretion of IL-6, IL-12 and NO (P <0.01), promote cell secretion of IL-4 and IL-10 (P <0.01), and the processing of the processing The post -effects are stronger.The results of the mouse break the hemostatic experiment show that: compared with the control group, the JXRPs group can speed up the hemostatic effect but there is no significant difference, and there is asignificant difference in the dosage of the JZJXRPs (P <0.05) in the medium dose (P <0.05), and the High doses have accelerated trend but there is no significant difference; The results of antioxidant activity in vitro showed that JXRPs and JZJXRPs had different degrees of scavenging ability for DPPH and ABTS, with IC50 values of JXRPs of 0.2215 mg/ml and 0.2110 mg/ml of JZXRPs, respectively, and IC50 values of 0.1651 mg/ml and 0.1884 mg/ml of JZJXRPs, respectively.Conclusion:After the Jiangxiangyu is produced by ginger juice, it can promote the dissolution of polysaccharides and increase the polysaccharide content. JZJXRPS anti -inflammatory hemostasis and antioxidant capacity are stronger than JXRPS, which can lay the foundation for the follow -up research and clinical application of JXRPS.
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Research progress of SHP-2 in tumor-associated macrophages

wuxueliang

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Abstract: Tumor-associated macrophages (TAMs) are the dominant cell group in the tumor microenvironment (TME) and are the most important regulatory cells of immune system suppression and tumor cell proliferation in Tmes. Src homologous 2 protein tyrosine phosphatase 2 (SHP-2) is a non-receptor protein tyrosine phosphatase that plays an important role in signaling from the cell surface to the nucleus and is a key intracellular regulatory factor mediating cell proliferation and differentiation, involved in a variety of growth factor and cytokine signaling pathways. Recent studies have shown that SHP-2 is a key enzyme that determines the function of tumor-associated macrophages, but because of its variable function, it plays different or even opposite roles in different solid tumor microenvironments. Based on this, this paper reviews the function of SHP-2 in TAMs and the role of ShP-2 in related solid tumors, providing a solid scientific basis for tumor immunity and targeted therapy.
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Antitumor mechanism of Ardisia Crenata

renqunli

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Ardisia Crenata Radix is a traditional Chinese medicine, which belongs to Myrsinaceae, and its main components are coumarins, saponins, flavonoids and volatile oil. Bergenin, ardisicrenoside A, ardisicrenoside B,ardisiacripin A,ardisiacripin B and embelin are active anticancer compounds with the in-depth study of the anti-tumor effect of Ardisia Crenata Radix. They show high potential in oral cancer, nasopharyngeal carcinoma, liver cancer, colon cancer, bladder cancer, cervical cancer and leukemia mainly through inducing tumor cell apoptosis, increasing tumor cytotoxicity, inhibiting cell proliferation, inhibiting tumor cell metastasis and migration, and inducing cell regulatory enzyme cascade reaction. However, the preclinical experimental data of cinnabar root anti-tumor mechanism are lack of high-quality, multi-sample and multi-repeated randomized controlled trials, and the clinical research data are lack of tumor prognosis, pharmacodynamics and pharmacokinetic data. Accurate research experiments and clinical trials should be designed to further explore the pharmacological effects of mining cinnabar root.
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Research progress in immunoregulation mechanism of exosomes on bacterial infection

李, Lizhiwei

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Exosomes are small endosomal derived extracellular vesicles with lipid bilayer structure, which contain substances such as protein, lipids, DNA, RNA, miRNA and lncRNA. Exosomes participate in pathogen recognition, antigen presentation, autophagy regulation, immune activation and immunosuppression in bacterial infection. Studies have shown that miRNA, lncRNA and protein in exosomes play important roles in regulating antibacterial reaction of organism. We reviewed the immunomodulatory effects of exosomes on several intracellular and extracellular bacterial infections to provide evidence for studying the interaction between exosomes and bacterial infections.
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Application and challenges for NHP laboratory animals in neuroscience

HuLingfeng, FanShengtao, LiuJie

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【Abstract】: Currently, non-human primates (NHPs) are becoming increasingly important in laboratory animals, especially in the field of neuroscience, where many significant breakthroughs have been made, including research on brain development, neurodegenerative diseases and psychiatric disorders, but as their breeding and use grows, biosafety and animal ethics issues should be taken into account. This review summarizes the application and challenges of NHP laboratory animal in the neuroscience field from the aspects of NHP overview, feeding and operation, biosafety, and animal ethics.
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Research progress on evaluation of type 2 diabetic peripheral neuropathy animal model

zhuziyue

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Diabetic peripheral neuropathy is a common diabetic complication. At the moment, our understanding of its pathogenesis is incomplete, and there are no effective treatment options. In-depth research requires the use of animal experiments. The criteria for modeling success and the evaluation method for peripheral nerve function recovery are critical for carrying out animal experiments on type 2 diabetic peripheral neuropathy, but there is a lack of sorting out and analyzing the evaluation methods for type 2 diabetic peripheral neuropathy models. As a result, the author reviewed recent data, summarized and analyzed the evaluation methods of animal models of type 2 diabetic peripheral neuropathy from small and large nerve fibers, and proposed future development directions in order to provide a reference for related research.
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A review and promising future directions of machine learning in laboratory animal facility management

xuxiao, Chen Qi

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As the intelligence level gradual improving in domestic laboratory animal facilities，a large amount of valuable data has been accumulated. The data has not been fully exploited due to lacking of analytical means. In the context of Big data, Machine learning algorithms have achieved remarkable results in biomedical, building science and other fields, and also provide a reference for their application in laboratory animal facility management. In this paper, the contents, methods and models of machine learning methods applied to various systems of laboratory animal facilities at home and abroad are reviewed and prospected.
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Optimization of the preparation method of lung tissue cryosections for multiplex immunofluorescence staining

Ye Qianchen, Xu Dan, Wen Fuqiang, Chen Jun, WANG TAO

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Objective Optimizing the preparation method of mouse lung cryosections to improve the quality of lung cryosections helps to enhance the specificity of immunofluorescence staining and obtain more accurate and reliable experimental results. Methods C57BL/6 mice were used to make cryosections by the traditional post-freezing fixation method, the pre-freezing fixation method, and a modified perfusion pre-freezing fixation method, respectively. A laser scanning confocal fluorescence microscope was used to observe lung tissue immunofluorescence staining. The whole areas of mouse lung slices were scanned by fluorescence microscope, and then the numbers of intact airways per unit area of lung tissue were calculated. Results For the lung cryosections made by the traditional post-freezing fixation method, the alveoli structure was damaged, and the airway wall was seriously broken with non-specific staining. The lung cryosections made by the pre-freezing fixation method showed relatively intact alveolar and airway structures but collapsed alveoli and several destroyed airways. The structure and morphology of the alveoli and airways were intact and clear in the lung cryosections prepared by the modified perfusion pre-freezing fixation method. In addition, the locations of target genes were accurate with multiple immunofluorescence staining. The number of intact airways (diameter ≥100 μm) per unit area in the lung cryosections from the modified perfusion pre-freezing fixation method was higher than from the pre-freezing fixation method (0.66±0.15 /mm2 vs. 0.33±0.14 /mm2, P<0.05), and was also significantly higher than that from the traditional post-freezing fixation method (0.66±0.15 /mm2 vs. 0.02±0.04 /mm2, P<0.01). Conclusions The modified perfusion pre-freezing fixation method is beneficial to maintain mouse lung tissue's morphological integrity and obtain high-quality multiplex immunofluorescence staining results.
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To explore the effects of different types of drinking water on the growth and fecal flora of mice

wuxuying

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【Abstract】 Objective to explore the effects of different types of drinking water on growth and fecal flora of mice. Methods SPF NIH mice were selected and divided into 5 groups with 32 males and females in each group. they were fed with pure water (control group), acidified water, alkalized water, weak acid water and solid water, respectively. diet and body weight were monitored continuously for 20 days. after the experiment, animal feces samples were collected, and the V3-V4 region was amplified with bacterial16S rDNA universal primers. Illumina Miseq high-throughput sequencing platform was used for high-throughput sequencing, and microbial community, alpha diversity and beta diversity were analyzed by bioinformatics method. Results in terms of body weight, the water of weak acid in female mice was higher than that in the control group at different pH values, while the other groups were lower than that in the control group(P﹥0.05). in male mice, the body weight of acidified water group was higher than that of control group, while that of other groups was lower than that of control group, but there was no statistical difference between groups(P﹥0.05). the body weight of female and male mice with solid water was lower than that of control group(P < 0.05). in terms of food intake and water intake, the water intake and water intake of female mice in the alkalized water group were lower than that of other groups, and the water intake of female and male mice in the solid water group was lower than that of other groups. through OTU cluster analysis, the dilution curve showed that the data volume of this sequencing was reasonable. the fecal flora of NIH mice was divided into five phyla, among which bacteroidetes and firmicutes were the main phyla. at the genus level, pseudopurpuromonas, lactobacillus and mycobacterium were the main genera. there were differences in fecal flora abundance and diversity among the five drinking water types. alpha analysis showed that the acidified water group had the highest flora abundance and diversity, while the solid water group had the lowest flora diversity. beta analysis showed that the fecal flora of the solid water group and the control group were the most similar, followed by alkalized water group, acidified water group and weak acid water group, respectively. Conclusions through the exploration of different types of drinking water, it is revealed that the solid form of drinking water has a great effect on the body weight, feed intake, water intake and fecal flora of mice, and the abundance and diversity of fecal flora of mice are affected by different pH values of drinking water, especially acidified water has a greater effect on the fecal flora of mice.
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The therapeutic effect of licorice zinc on chloasma mice was explored based on nrF-2 /HO-1 pathway

Du Xiaoshuang, Liu Ping, Deng Ying, Yang Hongqiu, Du Yu

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[Abstract] Objective To investigate the therapeutic effect of licorice zinc on melasma. Methods Melasma was induced by 100 mJ/cm2 UVB irradiation combined with 15 mg/kg progesterone injection. Tranexamic acid (0.065 g/kg) and low (0.65 g/kg)/medium (1.3 g/kg)/high (2.6 g/kg) zinc licorice were treated for 14 days. Skin was taken for HE and Masson-Fontana staining; and SOD, MDA, GSP-Px, TNF-α, IL-1β and IL-6 contents; plasma protein Nrf-2, nuclear protein Nrf-2, and HO-1 expression levels were measured. Results High dose of zinc licorice significantly reduced melanocyte formation, collagen cell necrosis and inflammatory infiltration, significantly up-regulated the expression of GSP-Px and SOD, decreased the expression of MDA, IL-6, IL-1β and TNF-α, up-regulated the expression of nuclear proteins Nrf-2 and HO-1, and down-regulated the expression of plasma protein Nrf-2. Conclusion Zinc licorice activates Nrf-2/HO-1 pathway to initiate high expression of HO-1, SOD and GSP-Px against oxidative stress, thereby reducing melanogenesis.
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Study on the mechanism of oral microflora involved in areca nut extract induced oral ulcer in rats

pengdongdong, chenxiangchi, tangzining, liuxuewu, yuanxiangzhong, limeng, liqiao, zhangzeheng

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Objective The model of oral ulcer in rats was constructed by areca nuts extract, and the structural changes and diversity characteristics of oral flora were observed, so as to explore the pathogenesis of oral flora involved in oral ulcer in rats induced by areca nuts extract, providing theoretical support for clinical prevention and treatment of oral ulcer. Methods Thirty SD rats were randomly divided into normal group, model group, drug treatment group, 10 rats/group. The rat oral mucosa was injected with 10g/mL areca nuts extract subcutaneously to replicate the rat oral ulcer model. Ulceration area, ulceration score, and levels of TNF-α, IL-2, and IL-8 in local tissues were observed, and HE staining was performed on oral mucosal tissues to observe the morphological changes of oral mucosal tissues. High-throughput sequencing method was used to analyze the structure distribution of oral microbial flora and microbial community diversity. Results Compared with normal group, the ulcer area of rats in model group was significantly increased (P<0.01), the ulcer score was significantly increased (P<0.01), the levels of IL-2, IL-8 and TNF-α in oral mucosal tissue were significantly increased (P<0.01), and the shannon index of oral saliva of rats was significantly increased (P<0.05).simpson index was significantly decreased (P<0.01), Strep-tococcus and Veillonella were significantly decreased (P<0.05,0.01), oral mucosal epithelial cell proliferation or focal necrosis, mucosal lamina propria edema and hemorrhage were observed in model group. It was accompanied by abundant infiltration of neutrophils and monocytes. Compared with model group, the ulcer area and ulcer score of model group were significantly decreased (P<0.05), the levels of IL-2, IL-8 and TNF-α in oral mucosal tissue were significantly decreased (P<0.05,0.01), and the shannon index of oral saliva of rats was significantly decreased (P<0.05). Simpson index was significantly increased (P<0.01), Strep-tococcus was significantly increased (P<0.001), Staphylococcus was significantly decreased (P<0.01), and the pathological degree of oral mucosa was significantly improved. Conclusion Areca nuts extract can successfully reproduce the rat model of oral ulcer, and it is speculated that the development of oral ulcer disease induced by areca nuts extract may be related to the imbalance of oral flora and local tissue inflammatory mediators.
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Effects of 5-Aza-CdR on hippocampal neurogenesis in mice via the Notch1 pathway

wangbaojie, suliqing, yanlei, zhangzhiyong, 王志广, baosuya, shaoguo

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【 Abstract】 Objective To investigate the effect of 5-Aza-CdR on Notch1 pathway and neural regeneration, and to explore the effects of 5-Aza-CdR on learning memory ability in mice by exploring active avoidance behavioral experiments on mice. Methods Sixty 6- to 8-week-old ICR male SPF-grade mice were divided into two groups, and 5-Aza-CdR was administered to one group of mice via lateral ventricular injection (i.c.v.), while the other group was injected with BSA as a control group. 5-Aza-CdR was administered to mice by lateral ventricular injection (i.c.v). mRNA and protein expression levels of Notch1 and HES1 were detected by real-time PCR and Western blot 24 hours after injection; 5-bromo-2'-deoxyuridine (BrdU)-positive cells were observed by laser confocal microscopy and Notch1 expression in hippocampal DG by laser confocal microscopy; Notch1 methylation changes were detected by MS-PCR; and learning and memory behaviours of mice were assessed by measuring passive avoidance tests and shuttle avoidance assays. RESULTS Injection of 5-Aza-CdR increased hippocampal Notch1 pathway activity and promoted neuronal regeneration in the DG region, decreased methylation levels in the Notch1 promoter region, and enhanced the ability of mice to perform active avoidance behaviour. Conclusion The effect of 5-Aza-CdR on active avoidance behavior may be related to the influence of hippocampal neural regeneration through the Notch1 pathway.
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Effects of emodin on autophagy and cycle arrest in fibroblast-like synoviocytes in rheumatoid arthritis

Tao Bonan, WANG Yonglan, HONG Lan, YUAN Lin

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Objective: Based on the cGAS/STING signaling pathway to investigate the potential effect of emodin (EMD) on autophagy of human rheumatoid arthritis fibroblast synovial cells (MH7A). Methods: CCK-8 method was used to detect the results of MH7A cell proliferation, and the concentration of drug was screened according to cell survival rate. Then, autophagy inhibitor 3-MA was added to further verify the effect of emodin on autophagy. Autophagy of MH7A cells was detected by MDC method. The protein expression levels of cGAS, STING, p-STING, LC3-I, LC3-II, p62 and Beclin-1 were detected by Western blot. Results: MDC staining indicated that emodin could enhance autophagy of MH7A cells. Western Blot results indicated that emodin could decrease the expressions of autophagy related proteins cGAS, STING, p-STING and P62, and increase the expressions of LC3-II and Beclin-1 in MH7A cells. After addition of autophagy inhibitor 3-MA, the expression of P62 protein in MH7A cells increased, while the expression of LC3-II and Beclin-1 protein decreased. Conclusion: Emodin may accelerate autophagy and inhibit MH7A cell proliferation by down-regulating cGAS/STING signaling pathway.
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Progression of the Relationship between Pyrozosis and Atherosclerosis

huangshuning

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Pyroptosis is a programmed mode of death, and activated aspartase-1 (caspase-1) can induce the occurrence of pyroptosis, promoting the release of inflammatory factors. In recent years, studies have found that pyroptosis may be closely related to the occurrence, development and outcome of atherosclerosis. This article reviews the relevant studies of endothelial cells pyroptosis, vascular smooth muscle cells pyroptosis and macrophages pyroptosis in the development of atherosclerosis, aiming to provide new ideas for the pathogenesis and diagnosis and treatment of atherosclerosis.
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Gli2 regulates the activation of Hedgehog pathway on proliferation, metastasis and epithelial mesenchymal transformation of Tca8113 cells

Liu Maolin, Wang Xiaotang, Song Xiaona, Ma Yunfei, Chang Xiaoqi, Song Guohua

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Objective: To investigate the effects of Gli2 on the proliferation, growth, migration and invasion of oral cancer cells (Tca8113) at the cellular level, and to clarify the molecular mechanism of Gli2 regulation on the migration and invasion of oral cancer cells. Methods: siRNA was used to inhibit Gli2 expression in Tca8113 cells. The effects of Gli2 on the proliferation, growth, migration and invasion of Tca8113 cells were examined by CCK-8, plate cloning and transwell chamber assay. Further, qRT-PCR and Western blot were used to explore the mechanism of Gli2 regulation of malignant proliferation and metastasis of Tca8113 cells. Results: The mRNA and protein expression of Gli2 in oral cancer cells (Tca8113) were increased. Interference with Gli2 expression inhibited the proliferation, growth, migration and invasion of Tca8113 cells. Further studies showed that interfering with Gli2 expression inhibited mRNA and protein expression of key factors in Hedgehog (Hh) pathway. In addition, interference with Gli2 expression can significantly affect mRNA and protein expression of key factors in Epithelial mesenchymal transformation (EMT) pathway. Conclusion: Gli2 is abnormally activated during oral cancer, and interference with Gli2 expression significantly inhibits the proliferation, growth, migration and invasion of oral cancer cells. Gli2 influences the migration and invasion of oral cancer cells by regulating Hh pathway and EMT pathway. This study provides a new way to elucidate the pathogenesis of oral cancer and a new perspective for clinical treatment of oral cancer.
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A model study of blood in urine (bladder cancer) under scorching humid fever under MNU-induced SD rats

changtuo, wangjie, linan, wangtianqi

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Objective: To investigate the modeling process and mold formation rate of N-methyl-N-nitrosourea (MNU)-induced lower jiao dampness-heat syndrome uremia bladder cancer (BC) model. Methods: Batch samples were collected by bladder perfusion (MNU) and H&E staining and pathological observation of pathological tissues were carried out at a total of 6 time points before, during and after molding, so as to understand the formation and development of carcinoma in situ of the bladder. Results: The experimental process was smooth, and no obvious urinary tract bleeding in rats occurred during the operation. By this modeling technique, considerable bladder tumors can be obtained after the eighth week, and there are obvious epithelial hyperplasia, damage and large-scale tumor formation in the bladder of rats in the model group. Conclusion: MNU can induce SD rat bladder cancer model with obvious model formation and low mortality, and the experimental data can provide a certain reference for the establishment, improvement and application of bladder cancer model.
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Protective effects of HIF-1α pathway in cold storage of isolated organs

wangzhiyang, zhengsiyang, zhuliang

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Organ transplantation is the main treatment for organ failure. The functional protection of donor organs during ex vivo transportation is the key to the success of organ transplantation. How to protect the functions of donor organs during in vitro transportation is one of the key issues in the field of organ transplantation research. In a hypoxic environment, the transcriptional activity of a series of genes in cells is activated. These genes are mainly involved in angiogenesis, iron metabolism, glucose metabolism, and cell proliferation/survival. In aerobic organisms, hypoxia-inducible factor-1α (HIF-1α) is involved in the regulation of the expression of various genes to maintain the homeostasis of tissues and cells under hypoxic conditions, thereby adapting to the hypoxia. Many studies have shown that the HIF-1α pathway plays an important role in protecting isolated organs from cold ischemia injury during cold storage. HIF-1α has been a hot spot in the research on the protective mechanism of cold ischemia injury of isolated organs. The intervention of the HIF-1α-related signal pathway is expected to become a new strategy for maintaining organ function during cold storage of isolated organs.
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Advances in animal models of diet-induced nonalcoholic fatty liver disease

Wang Jialin, Wang Chongyang, Li Yang, Chen Xianwei, Yuan Zhongnan, Zou Chaoxia, Zou Chendan

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The establishment of scientific and effective animal models is a key step to complete disease research. Diet induction is a common method to establish animal models of nonalcoholic fatty liver disease (NAFLD). The common animal models of diet-induced NAFLD mainly include high-fat and high-sugar diet-induced model, high-fat and high-cholesterol diet-induced model, and choline-deficient diet-induced model. Due to the different nutrients ingested by different modeling methods, the pathological characteristics such as fatty deformation, inflammation and fibrosis of the liver are different. In addition, different animal models vary in disease progression, disease severity, and applicable studies. This article analyzes and compares the common animal models of NAFLD induced by different diets in modeling methods, modeling time, pathological characteristics, applicable research, and related advantages and disadvantages, so as to provide reference for NAFLD researchers to select animal models.
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Isolation and extraction of Kupffer cells and hepatic stellate cells in mouse liver

mawenmeng, wuhao, yanyutong, sunxun, zhengqianqian

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【】Objective It is to explore the extraction and purification methods of Kupffer cells and hepatic stellate cells from mouse liver, and provide reference and suggestions for the separation and extraction methodology of primary non parenchymal cells from mouse liver.Methods Based on in vivo collagenase perfusion digestion, different reagents and methods such as Percoll and OptiPrep were used to extract C57BL/6 mice Kupffer cells and hepatic stellate cells and evaluate the purity through flow cytometry and immunofluorescence methods. Results The two-layer Percoll method for extracting Kupffer cells and the two-layer OptiPrep method for extracting hepatic stellate cells are feasible,and the purity can reach more than 90%.The cell yield was 1~2×107/liver, the cell survival rate was more than 90%.After 48 hours of primary cell culture, the number of Kupffer cells F4/80-positive cells and hepatic stellate cells α-SMA-positive cells reached more than 75%. Conclusions The separation and extraction method of Kupffer cells and hepatic stellate cells from mouse liver is perfect, reliable, cost-effective and reproducible.
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Application Research Progress of Single-cell RNA-sequencing Technology in Neurodegenerative Diseases

Wujianfei, Wangbinglong

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Neurodegenerative diseases (NDs) are the mental diseases closely related to the central nervous system, characterized by morphological abnormalities and slowly progressive loss of function in specific neuron groups. The main diseases include Alzheimer's disease (AD), Parkinson's disease (PD), Amyotrophic lateral sclerosis (ALS), Multiple sclerosis (MS) and Huntington's disease (HD). However, the directly therapies for NDs are not founded. In recent years, single-cell RNA-sequencing (scRNA-seq) technology has been widely used in various neurodegenerative diseases. It has been found that the pathogenesis of NDs are involved in conditions of immune cells and biological processes such as mitochondrial function, angiogenesis, inflammation and synaptic transmission. The antilepsis of induced pluripotent stem cells (iPSC) is a potential treatment for NDs. Ultimately, the application of scRNA-seq technology in various NDs was reviewed in this paper ,and provided reference for the future prevention and treatment of NDs.
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Research progress on modeling methods and evaluation indexes of kidney injury in zebrafish

lanailin, Huangchunhua, Xiaming, Limaojuan, Wangsiqi, Loudidong

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Renal function is to filter blood , regulate fluid and electrolyte balance, which is crucial for health of the body, moreover, the kidney is susceptible to toxicity of various compounds in and out of the body, resulting in renal function damage, hence the prevention and treatment of renal function injury caused by various factors is also a hot research topic. Zebrafish is an ideal animal model as it is closely related to humans in terms of morphology, physiology and function of kidney and its response to compounds. In this paper, we will review the methods and evaluation indexes of zebrafish kidney injury modeling.
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Microvascular morphometric analysis of cerebral cortical functional areas in C57BL/6 mice of different months of age

guomin

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Abstract: Objective To observe and analyze the different functional areas of cerebral cortex in C57BL/6 mice of different months. Methods Improved alkaline phosphatase staining was used to reveal the microvascular morphology of the cerebral cortex of C57BL/6 mice, including motor cortex (primary motor cortex, secondary motor cortex), sensory cortex (primary somatosensory cortex, secondary somatosensory cortex), visual cortex (primary visual cortex, secondary visual cortex), auditory cortex (primary auditory cortex, secondary auditory cortex), olfactory cortex (extrorhinal cortex, entorhinal cortex). Images were captured by OLMPUS BX51 microscope combined with Image-Pro Plus 5.1 software. The microvascular length density (Lv), microvascular surface area density (Sv) and microvascular volume density (Vv) were analyzed by Image-Pro Plus 5.1 software. Results The levels of alkaline phosphatase in microvessels of different functional areas of cerebral cortex increased with age and reached the peak in adults. There were four patterns for pia vessels to enter the cortex including T shape, Y shape, large arc, small arc. The Lv, Sv and Vv in different functional areas of the same aged mice showed a downtrend trend in the motor cortex, sensory cortex, visual cortex, auditory cortex and olfactory cortex. Moreover, the microvascular density of Lv, Sv and Vv in the motor cortex and sensory cortex was statistically significant compared with those in the olfactory cortex (P<0.05). The vascular densities of different functional areas in the aged mice were lower than those in the adult ones, but there was no statistical significance (P>0.05). Conclusion The improved alkaline phosphatase staining may clearly reveal the microvascular architecture in cerebral cortex of C57BL/6 mice and it provides morphological reference for the research of cerebrovascular diseases and the preparation of animal models.
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Research progress of traditional Chinese medicine for stroke based on epigenetic regulation

Fu Xue Qin

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Epigenetics is a branch of molecular biology, and is a research hotspot in the post-gene era in the field of life sciences. The application of epigenetic modification research methods is the frontier field of the pathogenesis of ischemic stroke, and it is the interpretation of traditional Chinese medicine in the treatment of ischemia. Targets of stroke and effective approaches to their mechanisms of action. At present, the etiology and pathogenesis of ischemic stroke has not been fully clarified, and modern medical treatment is still insufficient. The treatment of ischemic stroke with traditional Chinese medicine has unique advantages and broad prospects. The latest research has found that traditional Chinese medicine can participate in the regulation of abnormal epigenetic modification in the treatment of stroke. This article mainly starts from the epigenetic basis of traditional Chinese medicine theory of stroke and the application of DNA methylation, non-coding RNA and histone modification in stroke treatment and traditional Chinese medicine research, and explains the appearance of traditional Chinese medicine in the treatment of stroke. The role of heredity in order to provide new ideas and scientific basis for traditional Chinese medicine treatment of the disease
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Geniposide promotes skin ulcer wound healing in diabetic rats through PI3K/Akt pathway

chenxiaoyan

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Objective To investigate the protective effect of geniposide on diabetic skin ulcer and its mechanism. Methods Rats were divided into normal group, model group and geniposide subgroup (Gen-L: 200 mg/kg; Gen-H: 500 mg/kg). The diabetic rats were given normal saline or geniposide by intragastric administration (n = 6). Treatment was administered once a day, and the wound healing and inflammation of each group were recorded every day. After 7 days of treatment for diabetic skin ulcers, the wound area, tissue sections, TUNEL staining and Western blot were used to quantitatively analyze the changes of wound healing, apoptosis and related regulatory protein expression. Results Compared with the model group, geniposide (200 mg/kg and 500 mg/kg) orally could significantly promote the wound healing and increase the contraction of the injured area in diabetic rats. In the study of skin wound apoptosis in diabetic rats, TUNEL staining positive cells in geniposide subgroup were significantly reduced (P<0.05). Geniposide can significantly inhibit skin inflammation and promote wound repair, which may be related to the promotion of PI3K and Akt phosphorylation. Conclusion Geniposide can promote skin wound repair in diabetic rats by inhibiting inflammatory response and apoptosis.

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Volume 34,2024 Issue 10

Volume 34,2024 Issue 10