Abstract: Objective 　To establish and evaluate an animal model of rheumatoid arthritis with depression. Methods 　Thirty-two rats were randomly divided into normal group (N group), depression group (D group), arthritis group (RA group) and model group (RAD group), with eight rats in each group. The N group was fed normally, and the D group was fed under solitary rearing for 1 week and chronic unpredictable mild stress (CUMS) for 3 weeks. The RA group was subjected to collagen-induced arthritis (CIA) to establish the arthritis model. In the RAD group, the RAD animal model was established on the basis of the CIA animal model with 1 week of solitary rearing and 3 weeks of CUMS. Toe volume, arthritis index (AI), synovial and ankle joint pathology, and serum levels of interleukin-1β(IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were used to evaluate whether the rat model of RA was successfully established. The depression status of rats was evaluated by measuring body weight, food intake, sugar water preference rate, open field behavior (total number of motion cells and number of standing uprights), serum levels of corticotropin-releasing hormone ( CRH), adrenocorticotropic hormone ( ACTH), and corticosterone ( CORT), and pathological changes in the hippocampus. The RAD animal model was successfully established when both depression and RA were present in the rats. Results 　Compared with the N group, the toe volume, AI score, and serum levels of IL-1β and IL-6 in RA and RAD groups were significantly increased (P< 0. 05), joint space had widened, synovial hyperplasia was obvious and extended to the joint cavity, and a large number of inflammatory cells were seen. In D and RAD groups, body weight, food intake, total number of motion cells, number of standing uprights, and sugar water preference rate were decreased by various degrees, serum levels of CRH, ACTH, and CORT were increased by various degrees, and neuronal cells were reduced, accompanied by partial nuclear atrophy, especially in the model group (P< 0. 05). Conclusions CIA combined with solitary rearing and CUMS better simulates the changes in external manifestations and internal indicators of RAD model animals, which provides a reference to study RAD disease animal models.

Abstract: Objective 　To compare differences in chronic obstructive pulmonary disease (COPD) models induced by smoke inhalation through the nose-mouth plus lipopolysaccharide (LPS) or smoke exposure through the whole body plus LPS in rats, providing a new model for COPD. Methods 　Ninety male SD rats were randomly divided into normal control, whole body exposure, and smoke inhalation groups with 30 rats per group. The whole body exposure group was placed in a smoke box, whereby smoke contacted the whole body of the rats, whereas the smoke inhalation group inhaled smoke via the nose-mouth in a quantitative smoking device. Animals in both groups were exposed to smoke once a day for 60 min over 8 weeks. LPS (1 mg/ kg) was injected through the trachea on days 1, 7, 15, and 21 to induce the COPD model. Quality control of the smoke generated by the quantitative smoking device and smoking box included verification of the concentration stability and uniformity of the smoke particles and the particle size distribution of the smoke. At 4, 6, and 8 weeks of modeling, pulmonary function examination, the content of proinflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in alveolar lavage fluid, and histopathological examination were performed to compare the two modeling methods . Results 　Quantitative smoking devices produced smoke with a stable concentration of 1.1 mg/ L (total particles) and 0.1 mg/ L (nicotine), with a median mass particle size of 0.86 μm (nicotine) and a GSD of 2. 12. The deviation in stability and uniformity of the smoke concentration produced by the smoking box was obviously greater than that of the quantitative smoking device. Compared with the whole body exposure group, pulmonary function indexes FEV_0.2 / FVC and pulmonary compliance (Cdyn) in the smoke inhalation group were decreased significantly, and airway resistance (Penh) was increased significantly. IL-6 and TNF-α levels in alveolar lavage fluid of the smoke inhalation group were significantly increased at 6 weeks after modeling, and those in the whole body exposure group were increased at 8 weeks after modeling. The lesion severity of bronchial inflammation after modeling was similar in smoke inhalation and whole body exposure groups, but the degree of emphysema lesions in the nose-mouth inhalation group was more severe, and statistically significant differences in emphysema lesions appeared earlier in the nose-mouth inhalation group ( after 6 weeks of modeling) than in the whole body exposure group (after 8 weeks of modeling). The mean linear intercept (MLI) in the smoke inhalation group was increased significantly at 4 ~ 8 weeks of modeling, and the mean alveolar number (MAN) was decreased significantly at 6 ~ 8 weeks of modeling. MLI was increased significantly and MAN was decreased significantly in the whole body exposure group after 8 weeks of modeling. In the smoke inhalation group, significant abnormal changes were observed in pulmonary function indexes (FEV_0.2 / FVC, Cdyn, and Penh), cytokine levels in bronchoalveolar lavage fluid (IL-6 and TNF-α), and alveolar histopathological changes (bronchial severity and emphysema pathological score, MLI, and MAN) after modeling. However, the coefficient of variation (CV%) of each index was significantly lower than that in the whole body exposure group. Conclusion 　A 1. 0 mg/ kg LPS endotracheal drip combined with whole body smoke exposure or nose-mouth smoke inhalation both establish a typical rat COPD model. Inhaled smoke via the nose-mouth route shortened the modeling period. The model was established after 6 weeks of continuous smoke exposure, presenting typical symptoms of COPD (pulmonary ventilation dysfunction and broncho-lung chronic inflammatory infiltration accompanied by emphysema), and the difference between individual model animals was small (nose-mouth smoke inhalation vs smoke exposure, CV% values were smaller).

Abstract: Objective 　To investigate the effect of miR-185-5p on the proliferation, invasion, and migration of mouse breast cancer PY8119 cells. Methods 　miR-185-5p with significantly down-regulated expression in breast cancer was obtained through the preliminary research class; NCBI query human and mouse miR-185-5p gene sequences; EdU proliferation, Transwell invasion, scratch healing and flow cytometry were used to detect the effects of overexpression and knockdown of miR-185-5p on the proliferation, invasion, migration and apoptosis of PY8119 cells. The subcutaneous tumor model and lung metastasis model were established in C57BL/6 mice, and the changes of tumor proliferation ability and lung metastasis in vivo by overexpression of miR-185-5p were observed. Results 　EdU proliferation, Transwell invasion, scratch healing and flow cytometry assays showed that knocking down miR-185-5p significantly enhanced the proliferation, invasion, and migration of PY8119 cells and inhibited apoptosis. Overexpression of miR-185-5p inhibited the proliferation, invasion, and migration of PY8119 cells and promoted apoptosis. The in vivo tumorigenesis experiment in C57BL/6 mice showed that overexpression of miR-185-5p slowed down tumor growth. Lung metastasis analysis showed that overexpression of miR-185-5p inhibited lung metastasis in C57BL/6 mice. Conclusions 　miR-185-5p as a tumor suppressor gene inhibits the proliferation, invasion, and migration of mouse breast cancer PY8119 cells in vitro and in vivo.

Abstract: Objective 　To investigate the effect of perimenopausal depression on the expression of important genes in the Notch1 signaling pathway and DNA methylation of their promoters in the rat hippocampus from the perspective of DNA methylation of epigenetics. Methods 　Twelve-month-old female SD rats were randomly divided into control and model groups. Anxiety- and depression-like behavioral evaluations were conducted on rats using open field and sugar consumption tests, respectively. Real time quantitative PCR (RT-qPCR) was used to measure the expression of important genes in the Notch1 signaling pathway in the hippocampal dentate gyrus. Bisulfite sequencing was used to analyze methylation sites and levels in the promoter regions of important genes in the Notch1 signaling pathway. Results 　In perimenopausal depression rats, horizontal and vertical scores of the open field test and the rate of sucrose consumption were decreased significantly. Expression of important genes Jagged1, Notch1, and Hes5 in the Notch1 signaling pathway was significantly downregulated in the hippocampus of perimenopausal depression rats (P< 0. 01 orP< 0. 05). Some methylation sites and rates of the Hes5 gene were increased significantly, whereas the methylation sites and rates of Jagged1 and Notch1 genes were not significantly different (P> 0. 05). Conclusions 　The expression of important genes in the Notch1 signaling pathway in the hippocampal dentate gyrus of perimenopausal depression rats is significantly downregulated, and downregulation of the Hes5 gene may be related to the increase in the promoter DNA methylation level.

Abstract: Objective 　To establish a stable mouse model of mild-to-moderate closed head injury and evaluate its effectiveness. Methods 　A total of 170 SPF grade ICR mice were divided into a sham group (n= 34) and TBI group (n= 136). The TBI group was further divided into four subgroups, TBI-A, TBI-B, TBI-C, and TBI-D, with a drop height of 20 cm and drop weights of 60 g, 80 g, 100 g, and 120 g, respectively. According to the above conditions, the brain injury percussion was performed on the mice, 34 mice in each group. After modeling, the mNSS, fatigue rotating test, and Morris water maze test were used to assess balance and learning abilities of the mice. Brain water content analysis, evens blue staining, HE staining, GFAP immunohistochemistry, and TUNEL immunofluorescence were used to analyze pathological changes in brain tissue. Results 　Compared with the sham group, the mNSS and water maze escape latency in the TBI group increased to various degrees, whereas continuous walking time on the ratating rod decreased significantly, indicating severe neurological dysfunction in the TBI group. Brain water content and evens blue staining in brain tissue of TBI group mice were significantly higher than those in the sham group, suggesting blood-brain barrier damage and brain edema. Histopathological examination showed that neurons in brain tissue of the TBI group showed obvious degeneration and contraction, astrocyte proliferation, and the proportion of apoptotic cells was increased significantly. The damage degree increased with the increase in the weight of the impact. Comprehensive analysis revealed that weights of 60 g or 80 g with a height of injury at 20 cm simulated mild closed head injury in mice, whereas weights of 100 g or 120 g simulated moderate closed head injury in mice. Conclusions 　A stable mouse model of mild-to-moderate closed head injury was established using the principle of free fall, which lays the foundation to study the mechanism and treatment strategies for mild-tomoderate traumatic brain injury.

Abstract: Objective 　Bear bile powder (BBP), a natural Traditional Chinese medicine, has antioxidant and anticancer effects. To clarify the effect of BBP on the early period of hepatocarcinoma formation, we conducted anticancer efficacy experiments using short-term carcinogenesis bioassay models. Methods 　Forty 5-week-old male SD rats were randomly divided into DEN-alone, DEN + PBO, DEN + PBO + BBP-L (200 mg/ kg BBP), and DEN + PBO + BBP-H (400 mg/ kg BBP) groups, with 10 animals per group. All rats were injected intraperitoneally with 200 mg/ kg DEN, and the last three groups were provided a diet containing 0. 5% PBO. Additionally, rats of the last two groups were orally administered 200 or 400 mg/ kg BBP for 8 weeks. Results 　The relative and absolute liver weights of PBO-treated groups were significantly increased compared with those of the DEN alone group. However, the number and area of GST-P+ foci were significantly decreased in the DEN + PBO + BBP-L group compared with the DEN + PBO group. Additionally, the Ki-67+ cell ratio was significantly increased by DEN and PBO, and significantly decreased by BBP in both BBP-treated groups. Interestingly, the mRNA levels of Ccne1 and Ccne1b related to the cell cycle and Caspase 8 and Caspase 9 related to apoptosis were significantly increased in the DEN + PBO + BBP-H group. However, these changes were not observed in the DEN + PBO + BBP-L group. Conclusions 　BBP has a suppressive effect on the early period of hepatocarcinoma formation, leading to inhibition of preneoplastic lesions. Additionally, the suppression mechanism of BBP involves inhibition of cell proliferation and induction of apoptosis. A high dose of BBP may influence its inhibitory effect on preneoplastic lesions.

Abstract: Objective 　This paper uses bibliometric methods to reveal and visualize the current research status, hotspots and future trends of animal experiments on Alzheimer’s disease, in order to help researchers quickly understand the latest situation in this field. Methods 　We obtained publications in the field of AD animal experiments from 2018 to 2022 from the Web of Science Core Collection database. We used the analysis software CiteSpace 6. 2.R2 (Advanced) to assess the general information and keywords of publications in this field. Results 　After analyzing 4015 articles from 2018 to 2022, we found that the annual publication volume of studies had gradually increased. In terms of the number of articles, the United States, the University of California System, Saito T, and the International Journal of Molecular Science are the country, institution, author, and journal with the largest number of articles, respectively.“Alzheimer’s disease”had the highest frequency of occurrence, and“anxiety”had the highest central degree. Conclusions 　This field focuses on research on pathogenesis and treatment methods , but research is mostly confined to institutions and organizations, and there is a lack of international cooperation; in the future, more in-depth research should be carried out and international cooperation should be strengthened in order to promote the progress of research in this field.

Abstract:Objective 　To review the animal species, modeling method, and evaluation indexes of dilated cardiomyopathy (DCM) animal models at home and abroad to provide suggestions for improving the success rate of establishing DCM animal models with Chinese medicine characteristics, and to provide a reference to establish future DCM animal models. Methods 　The literature on animal models of dilated cardiomyopathy in the past 10 years was searched, and relevant reports were collected and summarized. Relevant parameters of experimental animals (species, sex, and body weight, et al), modeling method, modeling cycles, positive control drugs and cycles, and model evaluation indexes were summarized and analyzed. Results 　A total of 128 reports were included. Animal models of dilated cardiomyopathy were mostly prepared in SD and Wistar rats using anthracycline and adriamycin, with a cycle of 30 ~ 60 d. Echocardiography and pathological HE staining were usually applied to assess the success of model establishment. Among the studies containing positive control drugs, captopril was the most often used, followed by carvedilol. No studies involved explicit TCM evidence-based modeling. Conclusions 　Although many method are used to model dilated cardiomyopathy in animals, unified evaluation criteria are lacking. Therefore, it is recommended to integrate the result of imaging, pathology, biochemistry, and general conditions into the evaluation index of the model. Few studies on animals with frontier genetic genotypes exist in China, and standard dosages of modeling drugs for drug induction method are limited. Therefore, future modeling method and evaluation systems of dilated cardiomyopathy animal models need to be improved. Compared with animal models of heart failure, there is a lack of relevant evidence modeling, and the animal models of dilated cardiomyopathy in Chinese medicine require further development.

Abstract:On the basis of the compound factors of clinical practice, some progress has been made in acute exacerbation models of respiratory diseases by the two-hit method. In this review, we summarize the current research on acute exacerbation models of pulmonary fibrosis, chronic obstructive pulmonary disease, and bronchial asthma established by compound factors. The characteristics and application scope of each model are compared and analyzed in terms of animal selection, model preparation method, and major histopathological changes, providing a reference for researchers to further improve and perfect the model or rationally select animal models.

Abstract:Animal models of TCM are the experimental basis and necessary tools to conduct TCM research. Minipigs have stable genetic characteristics, a small size, easy breeding, docile nature, and similar physiology, biochemistry, and disease pathological manifestations to humans, which are ideal to establish experimental animal models and have broad application prospects and research value in the field of medicine. Researchers use small pigs to establish models of simple symptoms such as deficiency cold, blood stasis, actual heat, and spleen Qi deficiency. Models of atherosclerosis with spleen deficiency and phlegm turbidity and coronary heart disease with phlegm and phlegm stasis have achieved good result. However, the application of minipigs as TCM animal models is not widespread and worthy of further exploration.

Abstract:Animal models are widely used to study the pathogenesis of diseases and the safety, efficacy, and mechanisms of drug treatments. The selection and preparation of animal models that conform to the law of disease incidence are the basis and premise of experimental research. Diabetes macroangiopathy is characterized by extensive and rapidly progressing atherosclerosis of vascular walls. Mice are the most widely used experimental animals in atherosclerosis research. On the basis of the current research information and in accordance with the characteristics of type 1 and 2 diabetes, the development of diabetic atherosclerotic plaques, and the current monitoring technologies, this article systematically reviews diabetic atherosclerosis mouse models.

Abstract:Sex determination in fish is regulated by environmental and genetic factors, both of which can affect gonad differentiation and development. However, the way in which the environment affects gonad development-related genes to regulate the sex of fish remains unclear. Current studies suggest that high expression levels of the upstream genes sox9 and amh can induce primordium differentiation into spermatocytes in fish, sox9 promotes and amh inhibits spermatocyte proliferation, and elevated expression levels of the downstream genes ar and er can induce spermatocyte division, differentiation, proliferation, and spermatogenesis in fish. Exogenous estrogen inhibited the expression of sox9 and amh but induced ar and er at low concentration and inhibited them at high concentration. In conclusion, male sex differentiation in fish is regulated by sox9,amh, ar and er, and changes in environmental factors can affect the differentiation process. This report therefore reviews the upstream regulatory genes sox9 and amh and downstream regulatory genes ar and er and their relationships with sperm development. We also consider the effects of environmental factors such as stress on these genes and sperm structure, to explore sperm development and the gene-regulatory effects of environmental factors during this developmental process.

Abstract:It is widely believed that metacognition is a specialized cognitive capacity that only humans possess. A contentious debate in the field of comparative psychology is whether animals have metacognition. The exploration of this question will help researchers to uncover the phylogenetic roots of metacognition and to map the evolutionary history of higher cognitive functions in humans. This article reviews the main animal metacognition studies in the past 25 years. We summarize three common research paradigms and corresponding measures: uncertainty monitoring, information seeking, and confidence judgment paradigms. Two opposing explanatory tendencies and their rationales are outlined in the field of animal metacognition research: associative and metacognitive models. In response to the debate between associative and metacognitive models, two future research orientations are proposed to improve the research paradigm to assess the metacognitive model and to innovate theory to integrate these models into a new one.

Abstract:Systemic juvenile idiopathic arthritis (sJIA) is the most frequent rheumatic disease complicated by macrophage activation syndrome (MAS), which can be life-threatening in children. Current animal models show only some of the characteristics of sJIA and MAS, and there is no perfect animal model that can replicate the whole process of sJIA and MAS. In this paper, the clinical features, advantages and shortcomings of the currently proposed animal models of sJIA and MAS are reviewed, aiming to provide ideas for exploring better-represented animal models of sJIA and MAS.

Abstract:As an important intermediate link between in vitro cell experiments and clinical experiments, animal models are often preferred in researching tumor mechanisms, prevention, diagnosis, and treatment. The welfare of animals is closely related to the outcomes of animal experiments, serving as a crucial factor influencing the scientific validity and accuracy of research. Improving animal welfare can affect the reasonableness, feasibility, and ethical acceptability of animal experiments, thereby enhancing the reliability of research outcomes. This article summarizes the relevant aspects of experimental animals involved in tumor research, including tumor animal models, animal welfare related to tumor models, and humane endpoints.