Abstract: Objective To establish a myocardial infarction mouse model for systematic evaluation. Methods A total of 42 C57BL/ 6J mice were divided into two groups: sham group ( n= 8) and model group ( n= 34). Myocardial infarction was induced by squeezing the heart and left anterior descending coronary artery ligation. Echocardiography was performed to evaluate cardiac functions at 24 h, 3 d and 7 d after surgery. Then, the chest was opened and the occurrence of myocardial infarction was evaluated. Heart samples were collected for TTC staining to confirm the occurrence of infarction at 24 h after surgery. Heart sections were prepared for HE staining at 7 d after surgery. mRNA expression of inflammatory and fibrotic related genes in the infarct area was detected at 3 and 7 d after surgery, respectively. Results After surgery, the left ventricular anterior wall of the model group showed a whitish myocardium compared with the sham group. TTC staining showed a white infarct area, suggesting that the model was established successfully. Echocardiography indicated that cardiac functions of the model group were significantly decreased and gradually deteriorated over time. Expression of inflammatory genes, such as TNF-α and MCP-1, was increased significantly at 3 days postoperatively in the infarct area (P< 0. 05). Histological analysis showed that many inflammatory cells had infiltrated into the infarct area at 7 days postoperatively, and the boundary between the infarct area and non-infarct area was clear. Expression of fibrotic genes, including FN and COL3A1, in the infarct area were increased significantly ( P< 0. 01). Conclusions The MI mouse model and its evaluation were successfully established and refined, which provide insights for model establishment.

Abstract: Objective To establish an endothelial conditional connexin 43 (Cx43)-knockout mouse model and use it to examine vascular relaxation / constriction. Methods Eight Cx43flox / flox mice were crossed with C57BL/ 6 wild type (WT) mice and the offspring were backcrossed with C57BL/ 6 mice. This backcross process was repeated nine times, after which the Cx43flox / + mice were crossed with vascular endothelial-cell-specific Tie2-Cre recombinase mice to obtain Tie2-Cre /Cx43flox / + mice. The genotype was confirmed by mouse tail direct polymerase chain reaction (PCR). Expression of Cx43 in the superior mesenteric artery ( SMA) was measured by Western Blot and immunofluorescence. SMA rings were used to measure vascular relaxation and the contractile response to acetylcholine (ACh) and norepinephrine. Results Tie2-Cre /Cx43flox / +offspring were obtained and identified by PCR, Western Blot and immunofluorescence. Cx43 expression and AChinduced endothelium-dependent relaxation reactivity of the SMA were significantly decreased in Tie2-Cre / Cx43flox / + mice compared with WT mice ( P < 0. 01). Conclusions A mouse model with endothelium-specific Cx43-knockout was successfully established by intercrossing Tie2-Cre mice with Cx43^{flox / flox} mice. Cx43 expression in vascular tissues and the relaxation reactivity of SMA were decreased by conditional Cx43 knockout. These mice may thus provide an animal model for studying the link between Cx43 and vascular function.

Abstract: Objective To review the method used to prepare and evaluate animal models of perimenopausal depression, including animal selection, ovariectomy (OVX), chronic unpredictable mild stress (CUMS) stimulator use, modeling and treatment start time and behavioral testing. Methods We searched the CNKI and PubMed databases for studies of animal models of perimenopausal depression from the establishment of the databases to May 2022. Studies were screened according to defined inclusion and exclusion criteria, the relevant information was extracted and summarized, and charts were drawn. Results A total of 88 articles were identified, including 71 Chinese and 17 English articles. There were 65 rat models and 23 mouse models. The modeling method were OVX+CUMS and OVX+CUMS+solitary culture. Most studies in rats used SD rats (OVX), aged 90 ~ 120 days and weighing > 200 g; most studies in mice used KM mice, weighing about 20 g; 76% of the studies selected seven or eight stimulators, one or two of which were selected randomly each day and were not repeated within 3~ 7 days, with fasting, water deprivation, tail clipping, ice-water swimming, and day night reversal recommended method of simulation. Grouping according to behavioral result reduced the influence of individual differences. Vaginal smears were generally started 3 days after OVX and repeated for 5 days; 89% of the studies chose “treatment and modeling at the same time” or “ treatment after modeling”, and modeling and treatment should be consistent, generally 21 or 28 days. Positive control drugs included fluoxetine or fluoxetine combined with estrogen. The most common behavioral experiments were open field, sugar preference, and forced swimming experiments. Conclusions We reviewed the most widely used preparation method and evaluation indicators for animal models of perimenopausal depression, including details of the modeling processes, to improve the repeatability of preparing animal models of perimenopausal depression.

Abstract: Objective This article provides a reference for choosing a reasonable, reliable animal species,modeling method and evaluation index system, and provides a reliable animal model combining disease and syndrome for Chinese medicine syndrome differentiation. Methods We systematically searched the last 30 years for literature on animal models that combine a disease and a syndrome for six diseases related to diabetes mellitus, obesity, hyperlipoidemia, hypertension, hyperuricemia, and gout. Then we collected and organized the relevant literature, and summarized animalrelated parameters, modeling method, evidence types and evaluation systems. Results Among the 243 literatures retrieved, 132 literatures involved diabetes. The syndromes include deficiency of both qi and yin,damp heat trapped spleen type, internal heat due to yin deficiency, excessive internal heat; 3 literatures involved obesity. The syndromes include Spleen deficiency dampness resistance type; 9 literatures involved hyperlipidemia. The syndromes include Phlegm turbidity blocking type, qi stagnation and blood stasis type; 7 literatures involved hyperuricemia and gout. The syndromes include spleen deficiency phlegm dampness type, blood stasis type and damp heat accumulation type; 92 literatures involved hypertension. The syndromes include phlegm dampness essential, hyperactivity of the liver yang and vigorous liver fire type. Male rats (SD rat,Wistar rat) are involved in the most widely used experimental animals, quail is also applied to hyperuricemia and gout. The modeling method involved are mainly single-factor, dual-factor and hybrid-factor. The main evaluation method of creditial card combining disease and syndrome is to judge and characterize animal model by macroscopic symptoms, signs ( behavior experiments, mental status, livestock manure, tongue images, ect ) and pathological indicators ( glucolipid metabolism, serum biochemistry, liver and kidney function, ect). Conclusions The current modeling method and the main evaluation method of animal models that that combine a disease and a syndrome need to be improved to provide a reliable animal model combining diseases and syndromes for Chinese medicine syndrome differentiation and to establish conditions to study Chinese medicine syndrome differentiation.

Abstract: Objective To examine the potential mechanism of Ling Zhu Tusizi Pills in a rat model of adriamycininduced proteinuria. Methods Thirty-six specific-pathogen-free male SD rats were divided randomly into six groups (n= 6 rats per group), including blank, model, low-dose Ling Zhu Tusizi (14 g / (kg·d)), medium-dose Ling Zhu Tusizi (28g / (kg·d)), high-dose Ling Zhu Tusizi ( 56 g / ( kg·d)), and pyrrolidine dithiocarbamate ( PDTC; nuclear factor-κB inhibitor; 100 mg / (kg·d)) groups. The last five groups also received two injections of adriamycin into the tail vein to develop the nephropathy model, with one dose of 4 mg / kg and another dose of 2 mg / kg given 1 week apart. After successful establishment of the model, Ling Zhu Tusizi Pills were administered for 6 weeks. We examined general vital signs and body weights of the rats, as well as 24 h urine protein, plasma albumin, serum creatinine, blood urea nitrogen, and C-reactive protein. Morphological alterations of the renal tubules and glomeruli, renal fibrosis and basement membrane thickness were observed by hematoxylin and eosin, Masson, and periodic-acid Schiff staining. Modification of the foot process in glomerular podocytes was observed by transmission electron microscopy. Serum levels of pro-inflammatory molecules including interleukin (IL)-6 and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay. I-κB kinase β( IKKβ) / nuclear factor (NF)-κB/ monocyte chemoattractant protein-1 ( MCP-1) pathway-related proteins and nephrin and podocin proteins were detected by Western Blot. Results General health was poorer in the model group compared with the control group, as evidenced by body weight loss, increased 24 h urine protein, and decreased albumin. Renal tubule enlargement, disorganized glomerular visceral cell arrangement, diffuse foot process fusion, podocyte death,extensive fiber deposition, and basement membrane thickening were also noted in the model group. Serum levels of the proinflammatory markers IL-6, TNF-α and CRP were increased, renal-tissue phosphorylated (p)-IKKβ, p-NF-κB and MCP- 1 protein expression levels and the p-IKKβ/ IKKβ and p-NF-κB/ NF-κB ratios were increased, while nephrin and podocin expression levels were decreased ( P< 0. 05 ) in model compared with control rats. Rats in each treatment group experienced variable degrees of symptom relief compared with the model group, with medium-dose Ling Zhu Tusizi Pills having the best therapeutic effect. Rats in this group were generally healthy, with increased body weight and albumin and decreased 24 h urine protein. There were also notable reductions in fibrosis and renal tubule edema, a smooth arrangement of cells in the glomerular visceral layer, enhanced foot process fusion, and no thickening of the basement membrane. Serum levels of IL-6, TNF-α, and CRP were decreased, nephrin and podocin expression were increased, and p-IKKβ, p-NF-κB,MCP-1, p-IKKβ/ IKKβ and p-NF-κB/ NF-κB expression were all significantly decreased in renal tissues in this group (P<0. 05). Conclusions Ling Zhu Tusizi Pills reduced 24 h urine protein and serum pro-inflammatory markers in rats with adriamycin-induced nephropathy, possibly via inhibition of the IKKβ/ NF-κB/ MCP-1 signaling pathway.

Abstract: Objective Establish an animal model of small cerebral vascular disease (CVD) in SHR rats induced by D-galactose at various times. Methods Eighteen SHR rats were randomly divided into three groups according to body weight: D-galactose 150 mg / (kg·d) + 4 week, D-galactose 150 mg / (kg·d) + 8 week, and D-galactose 150 mg / (kg·d) + 12 week groups with six rats in each group. Another 6 WKY rats were used as the blank control group. A noninvasive sphygmomanometer was used to monitor blood pressure of rats every week during modeling. The Morris water maze was used to assess the cognitive function of rats after modeling. Brain, thymus, spleen, and liver indexes were measured by the weighing method . T-SOD, GSH-Px, MDA, NEFL and CALB/ SALB contents in rat serum were determined by ELISA. HE and LFB staining were used to observe cell morphology of the prefrontal cortex, ventricular microhemorrhage, and myelin sheath injury of the corpus callosum. Results Compared with the WKY group, the blood pressure of SHR rats was increased with the increase in D-galactose injection time. Learning and memory abilities were decreased significantly. Brain, thymus, spleen, and liver indexes were decreased. T-SOD and GSH-Px contents in serum were decreased, while MDA and NEFL, and CALB/ SALB levels were increased. The number of cytopathic lesions in the prefrontal cortex was increased, the amount of perivascular space and dorsal microbleeding of the third ventricle were increased, and vacuolization of the myelin sheath of the corpus callosum was increased. The most significant pathophysiological changes were observed in rats treated with 150 mg / ( kg·d) D-galactose for 12 weeks. Conclusions SHR rats injected with 150 mg / (kg·d) D-galactose for 12 weeks are a CSVD animal model similar to the human CSVD disease status.

Abstract: Objective To investigate miR-146b-5p expression in mice model of renal fibrosis induced by unilateral renal ureteral ligation, and to suppress miR-146b-5p expression to improve renal fibrosis induced by unilateral renal ureteral ligation in mice. Methods Twenty-four 8-week-old C57BL/ 6 male mice were randomly divided into sham operation group (sham), UUO model group (UUO), UUO+kidney miR-146b-5p knockdown group (UUO-KD), 8 mice in each group. In the sham group,the skin was only cut to expose and free the right kidney and ureter without ligation or disconnection. In the UUO group, the animal model of unilateral ureteral obstruction (UUO) was performed. In the UUOKD group, miR-146b-5p was specificly knocked down by electrotransferring the CRISPR/ RfxCas13 d plasmid in the mouse kidney. After 24 hours, the UUO mouse model was established according to the method of the model group, and the mice were sacrificed 7 days later to collect kidney samples. HE staining was used to observe renal pathological changes, Masson was used to detect the degree of renal interstitial fibrosis, immunohistochemistry was used to detect the expression of fibrosis-related proteins (α-SMA, FN, Col-1), and Western Blot and Real-time PCR were used to detect miR-146b-5p,α-SMA, FN, IL-1β, IL-6, TNF-α and other gene changes. Results Real-time PCR showed that miR-146b-5p was significantly increased in UUO model, and the gene was significantly decreased after electroporation knockdown of miR-146b-5p (P< 0. 05). Meanwhile,the expression of IL-1β, IL-6, TNF-α and other inflammatory factors was significantly down-regulated (P< 0. 05). It was observed after HE and Masson staining. Compared with the UUO group, the UUO-KD group had a better kidney structure, slightly deformed renal tubules, and less severe renal damage. The degree of fibrosis was significantly improved. And the results of immunohistochemistry showed that α-SMA, FN, Col-1 and other fibrosis indicators were also significantly reduced in the UUO-KD group ( P< 0. 0001). Conclusions Inhibition of highly expressed miR-146b-5p in UUO can significantly improve renal fibrosis, and miR-146b-5p may be a potential therapeutic target for renal fibrosis.

Abstract: Objective The aim of this study was to explore the mechanism of Yiqi Huoxue recipe in regulating immune inflammatory response in rats with coronary heart disease syndrome of qi deficiency and blood stasis. Methods Thirty SD rats were randomly divided into control group, model group, and recipe group. The model of qi deficiency and blood stasis syndrome of coronary heart disease was prepared by exhaustion, food control, and ligation of the left anterior descending branch of the coronary artery in model group, and recipe group. recipe group was pretreated with Yiqi Huoxue Recipe for 14 days from day 8 of modeling. On day 25, serum, heart, spleen, and aorta were collected for cytokine quantification and histopathological examination. Results (1)Serum levels of IL-37 and IL-35 were significantly lower in model group, than control group ( P< 0. 00001 ). ( 2 ) After treatment, the serum levels of IL-37 and IL-35 were significantly higher in recipe group than model group(P< 0. 00001). HE staining showed a large degree of inflammatory cell infiltration in the myocardial stroma in model group. The red pulp area of spleen tissue was smaller, and a large number of lymph nodes between marginal zone ( MZ) and the periarterial lymph sheath were seen in white pulp. Hyperplasia, structural disorder, the presence of foam cells, and lipid deposition were seen in the aortic intima in model group. Inflammatory cell infiltration of myocardial tissue was significantly lower in recipe group than in the model group. The intima of the aorta was thin, and scars were seen at damaged fibers. A normal distribution of trabecula(T)and splenic sinus(SS) was seen in spleen tissue and central antery(CA) morphology was regular in recipe group. ( 3) The average optical densities and protein contents of IL-35 and IL-37 in the myocardium, aorta, and spleen of model group were significantly lower than those in control group(P< 0. 00001). The average optical densities and protein contents of IL-35 and IL-37 in the myocardium, aorta, and spleen of rats in recipe group were significantly higher than those in control group (P< 0. 00001). Conclusions CHD model rats with qi deficiency and blood stasis syndrome support the theory of simultaneous treatment of heart and spleen, Yiqi Huoxue recipe improves the inflammatory state of CHD with qi deficiency and blood stasis syndrome, and its mechanism is related to the upregulation of IL-35 and IL-37.

Abstract: Objective To investigate the effect of an MST1 / 2 kinase inhibitor on expression of YAP, GAP43,Caspase 3, and GFAP in rats with acute spinal cord contusion ( SCC). Methods A total of 105 adult female SD rats weighing 180 ~ 200 g were used, of which 35 rats were subjected to a laminectomy and no drug injection as the sham operation group, whereas 70 rats were subjected to spinal cord contusion and divided into normal saline and MST1 / 2 inhibitor groups injected with normal saline or 1 mg / kg XMU-MP-1, respectively. Western Blot and immunofluorescence were used to observe YAP, GAP43, Caspase3 and GFAP expression. BBB and inclined plane tests were used to observe motor functions in rats. Results The inclined plane test and BBB scoring showed the scores of the MST1 / 2 inhibitor group were significantly better than those of normal saline group from day 7 to 28 (P< 0. 05). Western Blot showed that YAP expression in the MST1 / 2 inhibitor group was significantly higher than that the normal saline group from day 14. GFAP and Caspase3 expression in the MST1 / 2 inhibitor group was significantly lower than that in the normal saline group from day 14.GAP43 expression was also observed in the MST1 / 2 inhibitor group from day 14. These differences were statistically significant (P< 0. 05). Immunofluorescence showed that the MST1 / 2 inhibitor group had less infiltration of inflammatory cells and formed a structural framework of nerve tissue, whereas the normal saline group had obvious infiltration of inflammatory cells and formed a large number of glial scars. Immunofluorescence also showed that YAP was expressed in nerve cells in the Sham operation group. Moreover, the number of YAP and GAP43-positive cells in the MST1 / 2 inhibitor group was significantly higher than that in the other two groups, and the number of mature and hypertrophic astrocyte cells in the MST1 / 2 inhibitor group was significantly lower than that in the normal saline group; double immunofluorescent staining showed that YAP and GFAP were co-expressed. Conclusions A high dose of the MST1 / 2 inhibitor decreased YAP expression, activated reactive astrocytes, reduced the inflammatory injury response, inhibited apoptosis of nerve cells, improved the microenvironment of the injured area, and promoted neurite regeneration and function recovery.

Abstract:Calcium ions are important messengers in the nervous system, which produce various cellular signals and play an important role in the regulation of neuronal excitability. Calcium ions in the clear cell area perform highly specific functions. Calcium signals indirectly reflect the activity of neurons, calcium signal detection in neurons is especially important to study cerebral cortex functions. Two-photon calcium imaging in the cerebral cortex has a unique advantage by revealing real-time in vivo activity of cortical neurons at the single cell resolution. This article reviews the application of two-photon calcium imaging in the study of cortical functions in non-human primates.

Abstract:Acute kidney injury (AKI) is a common disease in clinical practice, and its incidence is increasing yearly along with a high mortality rate. Therefore, establishment of reliable, stable and reproducible AKI models is the basis to study kidney diseases. AKI modeling method are diverse in accordance with different experimental purposes. In this review, we describe the approaches to establish AKI models, namely drugs, unilateral ureteral obstruction, ischemiareperfusion, 5 / 6 nephrectomy, and infection. The purpose of this review is to discuss and summarize the current status and method to establish various AKI models, and to discuss the advantages and disadvantages of the induction method and their corresponding clinical signs in terms of their pathological mechanisms. This review provides a foundation for standardization of AKI animal models and the theoretical principles and experimental basis for in vivo studies of kidney diseases.

Abstract:Zebrafish has become a widely used model organism to study vertebrate development, genetic mechanisms of diseases, and drug toxicology. There is high similarity in the morphology, physiology, functions, and gene expression of zebrafish and human eyes. Furthermore, zebrafish embryos are easy to obtain, develop rapidly, and easily observed and manipulated. Therefore, zebrafish has become a good animal model for human ophthalmic diseases and provide new insights to further explore the pathogenesis of diseases and treatment method . This review introduces the ocular characteristics of zebrafish, the application of zebrafish animal models in ophthalmic diseases, and evaluation of ophthalmic drug efficacy and ocular toxicity of drugs.

Abstract:Atherosclerosis (AS) is a chronic inflammatory metabolic disease closely associated with risk factors such as hypertension, hyperlipidemia, hyperglycemia, and hyperuricemia. The intestinal barrier function plays an important regulatory role in maintaining intestinal homeostasis, and intestinal flora is an important component. Disturbance of intestinal flora can lead to intestinal barrier damage and an increase of intestinal permeability. The related products, such as lipopolysaccharide, short chain fatty acids, bile acids, and trimethylamine oxide change and can cause inflammation, oxidative stress, and other adverse reactions that then affect the occurrence and development of AS. In this article, we review the interaction between intestinal flora and the intestinal barrier in AS development and its risk factors, and provide a reference for the study and prevention of AS.

Abstract:Tooth development involves reciprocal interactions between oral epithelium and mesenchyme, and is achieved through complex regulatory networks. Abnormal spatiotemporal expression of the molecules involved in regulating tooth development may lead to abnormities of tooth development initiation and successional tooth replacement, which in turn affect the number of teeth,resulting in tooth agenesis and hyperplasia. In addition to using traditional mouse models, in recent years, animals such as miniature pigs, monkeys, and ferrets have been increasingly used in tooth development research. Here, we summarize tooth development models and molecular regulatory networks of vertebrate successional teeth to systematically understand the causes of abnormalities of the tooth number in humans.

Abstract:Human respiratory syncytial virus ( hRSV) is a major cause of respiratory disease and infant hospitalization worldwide. At present, there is no effective hRSV vaccine or therapeutic drug. A candidate vaccine for the hRSV susceptible population is currently being developed, and an animal model of hRSV would therefore play an important role in the preclinical trials of a hRSV candidate vaccine. Although many models have shown effectiveness in preclinical studies, few have entered clinical trials and their success rate has been low. Animal models to date have been unable to fully reproduce the pathogenesis of hRSV infection. Here, we analyze the advantages and limitations of hRSV animal models, including the use of hRSV to infect non-human mammalian hosts and the development of non-human pulmonary virus animal models. The aim of this review is to provide a rational strategy for the construction of an effective animal model of hRSV infection.

Abstract:This review summarizes the research progress of joint cartilage pathogenesis in hemophilia patients to further understand the unique mechanism of the development of cartilage damage to the occurrence of blood friends, and aims to provide new ideas to repair cartilage. English language databases, such as PubMed, Web of Science, SpringerLink, Google Scholar and Chinese language databases, such as CNKI, VIP and Wanfang, were selected. The English search key words were hemophilic, Joint bleeding, cartilage, Pathogenesis, cytokine, immunocyte, iron, synovitis, blood, plasma and signaling pathway. The Chinese search key words were hemophilia, joint hemorrhage, cartilage, pathogenesis, cytokines, immune cells, iron, synovitis, blood, plasmin and signal pathway. In accordance with the inclusion and exclusion criteria, literature with a high correlation of joint soft bone pathophysiology of hemophilia from January 1981 to March 2022 were searched, older or repetitive literature was excluded, and 60 related articles were finally included. and Conclusion Various cytokines, immune cells, iron ion deposition, chronic synovitis, blood, plasmin, and signaling pathways lead to chondrocyte apoptosis, leading to hemophilia cartilage injury in which iron ion deposition is critical for hemophiliac cartilage injury. Various influencing factors are interrelated and complex. How to regulate the damage of various influencing factors in articular cartilage is expected to become an important method to repair cartilage.

Abstract:Diabetic osteoporosis (DOP) is a chronic bone metabolic disease induced by diabetes mellitus, which is characterized by a decreased bone mass, increased bone fragility, decreased strength, and fracture tendency. To better explore the pathogenesis of DOP and develop treatments, it is important to establish an animal model that mimics the pathology of human DOP. The method of establishing a DOP animal model induced by streptozotocin has the advantages of simple operation, short time, high molding rate and relatively high stability. This article reviews animal selection, feeding,streptozotocin application, modeling method, modeling criteria, and testing indexes to provide a basis for more in-depth research of DOP pathogenesis.