Abstract:Objective We aimed to evaluate the immunotoxicity of type I hypersensitivity reaction of 5- hydroxymethylfurfural (5-HMF, C3H6O3 ) and its dimer 5,5′-dimethyl oxide (2-furfural) (OMBF, C12H10O5 ) in vivo and in vitro and to explore the mechanisms of action. Methods The in vivo experiments were performed on male Brown Norway (BN) rats, which were randomly divided into seven groups before a week of environmental adaptation. The groups were saline control (NS), adjuvant control (Adj.), model group (Model), low dose of 5-HMF (5-HMF-L) and high dose of 5- HMF (5-HMF-H), low dose of OMBF (OMBF-L) and high dose of OMBF (OMBF-H). BN rats in each group were given saline, adjuvant or adjuvant suspension of the corresponding concentration of drug, respectively. After 30 minutes reaction, blood was collected from the abdominal aorta, and the concentrations of IgE, IgG and IL-4 in the serum were determined by enzyme linked immunosorbent assay ( ELISA). Rat basophil leukemia ( RBL)-2H3 cells were used for the in vitro experiments. Cells were randomly divided into seven groups, as per the animal experiments. The degranulation of cells was observed by toluidine blue staining before and after administration of different concentrations of drug. The release rates of β- hexosaminidase (β-HEX) and histamine (His) in the supernatant of the cells were determined by chemiluminescence after drug administration. The mechanism of 5-HMF and its dimer OMBF-induced type I hypersensitivity was explored by Western blot analysis. Results Type I hypersensitivity can be induced by 5-HMF and its dimer OMBF at both low and high doses. The in vivo experiments demonstrated that there existed significant differences between the NS and all other groups in IgE, IgG and IL-4 in the serum, except IgG in the low dose 5-HMF group. In vitro experimental studies indicated that both the low and high dose groups showed cell degranulation, and the indicators measured showed different degrees of increase. The toluidine blue staining result showed that the degranulation of RBL-2H3 cells under the high dose of OMBF was more severe. The result of the β-HEX assay showed that the release rate of β-HEX from the supernatant of the high- dose OMBF group was higher than that of 5-HMF. The release rate of His was significantly increased when compared with the blank groups, but showed no major differences among the drug-administered groups. From the result of mitogen- activated protein kinase (MAPK) family phosphorylation levels, OMBF can promote the phosphorylation of MAPK more dramatically when compared with 5-HMF, especially in the expression of p-p38. These findings indicated that the immunotoxicity of OMBF was higher than that of 5-HMF. Conclusions Through in vitro and in vivo experimental models, we found that both 5-HMF and OMBF can induce type I hypersensitivity reactions as small molecule allergens. The levels of phosphorylated MAPKs were up-regulated, indicating that the immunotoxicity of 5-HMF and OMBF might be related to MAPK family proteins.

Abstract:Objective To investigate the metabolites of scopoletin using a rat liver microsomal incubation system and the effects of drug-containing serum on rat hepatic stellate cells (HSC). Methods The metabolites of scopoletin from the rat liver microsomal incubation system were analyzed by high-performance liquid chromatography-mass spectrometry (HPLC-MS) and the effects of drug-containing serum on rat HSC were observed. The metabolites of scopoletin were analyzed by high-resolution second-order mass spectrometry with an ACQUITY UPLC HSS T3 column (100 mm×2. 1 mm i. d.d., 1. 8 μm) and 0. 1% formic acid aqueous solution-0. 1% formic acid acetonitrile mobile phase. Annexin V-APC single staining and MTT method were used to observe the induction of apoptosis and inhibition of proliferation of HSC induced by drug-containing serum. Results Three metabolites of scopolactone were identified with the rat liver microsome incubation system, which indicated that the main metabolic pathway was demethylation and glucuronidation. The drug-containing serum had a weak effect on inducing HSC apoptosis and inhibiting HSC proliferation. Conclusions Combined with previous studies, the anti-fibrotic effects of scopolactone do not directly affect HSC, and further study is needed to clarify the possible mechanism.

Abstract:Objective To investigate single dose and repetitive dose toxicity of compound Yizhihao particles on pre-weaning Sprague Dawley ( SD) rats ( postnatal day 15; PND15 ) and provide a reference for human clinical use. Methods Five litters of female rats and their offspring were selected that met the requirements of the fostering design. Three litters were randomly divided into solvent control,compound Yizhihao particles dosage groups (70. 4, 88. 0 g / kg), according to the serial numbers of the female rats. A detailed observation of the rat, including examination of viscera morphology by naked eye, was conducted until day 14 post-drug administration. Simultaneously, 20 female rats and their offspring were selected that met the requirements of the fostering design. Each litter was randomly divided into solvent control, lcompound Yizhihao particles dosage groups ( 17. 6, 44. 0, 70. 4 g / kg) according to the serial numbers of the female rats. At PND₁₅ , compound Yizhihao particles was orally administered daily to the juvenile rats for 28 days, with recovery for 28 days after cessation. Parameters were examined including general condition, development index, ophthalmic examination, behavioral test, and histopathological examination. Results In the single dose toxicity study, three animals died in the 88. 0 g / kg dose group at day 2 while the weights of both the 70. 4, 88. 0 g / kg dose groups decreased significantly. In the repetitive dose toxicity study, compared with the solvent control group, the weights of both male and female rats in the 70. 4 g / kg dose group were significantly decreased at day 3. After 28 days of treatment, a decrease in mean crown-rump length and tibia length were noted in the males of the 70. 4 g / kg dose group. Urine bilirubin showed increases in the 44. 0, 70. 4 g / kg dose groups. The ratio of liver and kidney weight to body weight and to brain weight in the 17. 6, 44. 0, 70. 4 g / kg dose groups for both male and female rats were higher than those in the solvent control group. Conclusions The maximum tolerated dose of compound Yizhihao particles in juvenile rats was 70. 4 g / kg in a single dose toxicity study. The no observed adverse effect level was 44. 0 g / kg in a 28 day repetitive dose toxicity study.

Abstract:Objective To compare the changes in reproductive and developmental indexes after administration of different doses of cyclophosphamide before mating in Sprague Dawley ( SD) female rats, and to establish a standardized positive model for fertility and early embryo developmental toxicity test (Phase I). Methods One hundred and fifty SD rats (75 male, 75 female) were randomly divided into three groups ( n = 50; 25 male, 25 female), solvent control, cyclophosphamide 100 mg / kg and cyclophosphamide 20 mg / kg. Female rats were injected intraperitoneally with 20 mg / kg or 100 mg / kg of cyclophosphamide once daily in the 14 days preceding the mating period.The solvent control group was given the same volume of normal saline via the same route. Drug was administered from 14 days before mating until the 7th day of gestation (GD₇, the day found the sperm orthe pudendal embolism seemed as GD₀ ). No drug was administered to male rats in any group. General physical examinationof the animals was performed daily, body weight was measured twice a week and food intake was measured once a week. Female rats were sacrificed at GD₁₄ and pregnancy outcomes including pregnancy rate, loss rate before implantation, rate of implantation, the numbers of corpora lutea, loss rate after implantation, live fetuses, rate of live fetuses, uterine plus fetal weight and rate of absorbed fetuses were recorded. Results Compared with the solvent control group, the body weight and food intake of the two cyclophosphamide groups were all significantly decreased (P<0. 05 and P<0. 01).There was no significant difference in pregnancy, pre-implantation loss and implantation rates, or the average number of corpora lutea and the mean implantation number.The weight of uterus plus fetuses in the cyclophosphamide 20 mg / kggroup was significantly reduced (P<0. 01). The average number of live fetuses, the rate of live fetuses, and uterus plus fetuses weight were significantly reduced, and the rate of absorbed fetuses was significantly higher in both treatment groups compared with the control group (P<0. 05 and P<0. 01). Conclusions One hundred milligrams per kilogram and 20 mg / kg of cyclophosphamide administered intraperitoneally to female rats once and 5 times a day during the 14 days preceding mating, can successfully establish a standardized positive model for fertility and early embryo developmental toxicity in SD female rats, with 20 mg / kg cyclophosphamide representing the best option.

Abstract:Objective To establish a method to verify the performance of a flow cytometer under GLP conditions. Methods Based on the industry standard document, YY/ T 0588 - 2017 flow cytometer, a verification method for performance evaluation of a flow cytometer was established using a BD FACSVerseTM flow cytometer, including the commonly used flow detection settings for immunological assessment in drug safety evaluation, forward angular scattering light and lateral angular scattering light resolution, repeatability, carrying pollution rateand instrument stability, among others. Results Under the test conditionsin our laboratory, the performance parameters of the BD FACSVerseTM flow cytometer were in accordance with the technical requirements of the industry standard. Conclusions The above method is accurate and reliable, and can meet the needs of drug safety evaluation. It can provide a useful reference for the performance verification of flow cytometer under GLP conditions.

Abstract:Objective According to the technical guidelines for non-clinical safety evaluation of pediatric drugs by both domestic and foreign drug regulatory authorities, this study regarding repeated dose toxicity of compound Taraxacum mongolicum extract (CTME) in juvenile rats was performed to provide reference for clinical trials. Methods One hundred and forty-four juvenile Sprague-Dawley rats (postnatal day(PND) 27) were randomly divided into four groups with 18 of each gender per group. The rats were orally administered twice daily with CTME at doses of 0, 1. 2, 3. 8, 12 g / kg for 4 weeks in a dosage volume of 1 mL/ 100 g. Eighty rats were necropsied after the last administration, and the remaining animals were observed for 4 weeks after drug cessation. Test indicators included routine toxicology indexes, growth development, skeletal development, behavioral testing, sex hormones and immune system development. Results In the high dose group, the male juvenile rats slowly grew, reduced food intake and the juvenile rats of both genders demonstrated salivation, some changes in hematology ( decreased red blood cell count and hemoglobin levels, increased numbers and percentage of reticulocytes ), biochemistry ( decreased total carbohydrates, triglycerides and glucose levels ), urine (increased ketones, protein, leukocytes, specific gravity, pH and microalbumin), immunology ( increased spleen weight and coefficient, and increased IgG in male rats), mild extramedullary hematopoiesis in spleen and liver, and mild proliferation of erythroid hematopoietic cells in bone marrow. The influence could be reversed after 4 weeks of withdrawal. There were no obvious toxic effects on sexual development, skeletal development, growth hormone, behavior, memory, ophthalmology, sex hormones, or bone marrow cell morphology. No significant toxic effects were observed in the middle or low dose groups. Conclusions Under these test conditions, the no observed adverse effect level (NOAEL) of CTME in juvenile rats ( PND27 ~ PND54) was 3. 8 g / kg. However, a dose of 12 g / kg resulted in reversible changes in some indexes, with red blood cells being the main toxicity target.

Abstract:Objective To investigate and compare the angiogenesis ability and transplantation safety of mesenchymal stem cells derived from umbilical artery, umbilical vein and Wharton’ s jelly. Methods We isolated and cultured human umbilical cord artery perivascular stem cells (UCA-PSCs), umbilical cord vein perivascular stem cells (UCV-PSCs) and Wharton’ s jelly mesenchymal stem cells ( WJ-MSCs ). Flow cytometry was used to analyze the expression of cell surface markers at passage 3. The multipotency of cells was analyzed by adipogenic, osteogenic and neural-like differentiation assays. Immunofluorescence was used to detect the perivascular stem cell markers. Angiogenesis ability was analyzed by tube formation assay. Stem cells were transplanted into nonobese diabetic / severe combined immunodeficiency ( NOD-SCID) mice to test the biological safety. Results UCA-PSCs, UCV-PSCs and WJ-MSCs expressed mesenchymal stem cell markers that showed multi-lineage differentiation potential. UCA-PSCs expressed perivascular stem cell markers and had stronger angiogenesis ability in vitro than UCV-PSCs and WJ-MSCs. At 3 months after cell transplantation, hematoxylin and eosin staining showed no obvious morphological damage or abnormality in the organs. Conclusions UCA-PSCs had stronger angiogenesis ability than UCV-PSCs and WJ-MSCs. The transplantation of three kinds of stem cells into NOD-SCID mice result ed in no tumor formation, and appeared to be safe and feasible.

Abstract:Objective The pharmacokinetics of cellular drugs were studied with the mouse umbilical cord mesenchymal stem cells ( mUCMSC) tracing method and mdx mice. Methods mUCMSC were labeled by GFP transfection, and a 0. 4 mL (5×10⁶ GFP-mUCMSC) cell suspension was injected locally into the abdominal cavity of mdx mice with damaged gastrocnemius and diaphragma. The fluorescence signal in the mice was observed with a live imaging system at five timepoints including 1 h, 3 h, 5 h, 24 h, and 1 wk post injection. The mice were euthanized and their diaphragms were collected to obtain frozen sections for fluorescence microscopy. Results mUCMSCs were successfully labeled when they were co-cultured with the viral vector at an MOI of 150 for 72 h. One hour after local intraperitoneal injection of the GFP-mUCMSCs, a strong green fluorescent signal was observed with the live imaging system under the injection position on the abdominal wall of the mdx mice, and the fluorescent area gradually decreased with time. After 24 h, the fluorescence intensity was obviously weakened, and the fluorescent area was greatly reduced. These fluorescent signals had almost disappeared after one week. At this time, the fluorescent signals could still be observed in the diaphragm of the mdx mice under the fluorescence microscope, and this signal was significantly higher than that of the control group without cell injection (P<0.05). Conclusions Umbilical cord mesenchymal stem cells injected into the abdominal cavity of the mdx mice entered the injured diaphragm and their GFP fluorescent signals gradually disappeared within 1 week. Thus, a live imaging system is an effective tool for monitoring the dynamic distribution and clearance rate of stem cells in vivo.

Abstract:Objective To establish a stably expressing human GPC3 SK-Hep-1 cell line. Methods The eukaryotic expression vector pcDNA3. 1-GPC3 was constructed and transfected into SK-Hep-1 cells by electroporation. The G418 optimum screening concentration for SK-Hep-1 cells was 700 μg / mL. Transfected cells with the target gene were screened with G418 for 20 days. Analysis of GPC3 expression in the stable cell line was performed by Western blot and flow cytometry. Results Western blot result showed that the GPC3 expressing level of SK-Hep-1/ GPC3 was markedly higher than that of HepG2. Flow cytometry demonstrated GPC3 protein on the surface of SK-Hep-1/ GPC3 cells. Conclusions SK-Hep-1/ GPC3 cells that stably expressed GPC3 protein were successfully established. These result provide a solid foundation for further research on GPC3 therapeutic antibodies.

Abstract:Objective Differences in peripheral blood and immune cell phenotype were compared between aged and young Sprague-Dawley rats. Methods The peripheral blood cell count, leukocyte classification, immune organ coefficient, immune cell phenotype in peripheral blood and spleen, and p16 expression in spleen T cells were detected in 8-week-and 12-month-old rats. Results Compared with young rats, the peripheral blood leukocyte count and percentage of lymphocytes in aged rats were decreased, and the percentages of erythrocytes, hemoglobin, neutrophils, eosinophils, and basophils were increased. However, there was no difference in the percentages of platelets and monocytes between groups. Additionally, there were no differences in thymus coefficient and spleen coefficient between young and old rats. Peripheral blood immunophenotyping indicated helper T cells, regulatory T cells, natural killer cells, and monocytes were increased, whereas B cells were decreased in aged rats compared with young rats. However, there was no significant difference in cytotoxic T cells. In old rats, markedly lower numbers of B cells (CD45RA) were present in the spleen compared with young rats, and there was no significant difference in CD3 cells between the groups. In addition, the expression of P16mRNA in spleen T cells of aged male rats was significantly higher than in young male rats. Conclusions In aged rats, the differentiation of peripheral blood cells was deviated, the proportion of neutrophils was increased, and the proportion of lymphocytes was decreased compared with young rats. The proportion of regulatory T cells in the peripheral blood was increased, and the proportion of B lymphocytes was decreased in the peripheral blood and spleen in aged rats compared with young rats. These changes in innate immunity and acquired immunophenotype suggest that the immune function of aged rats was decreased and the regulation of immune function was disordered, similar to changes in the immune system of elderly population. The expression of P16 in spleen T cells was increased in aged rats, and therefore might be used as a biomarker for immune senescence. This study provides primary data for the study of age-related diseases and ageing animal models.

Abstract:Objective To genotype clinical strains of Pasteurella multocida isolated from conventional rabbits bred in different laboratory animal facilities. Methods From 2000 to 2018, 14 strains of P. multocida were isolated from laboratory rabbits obtained from different laboratory animal facilities; these were then cultured and identified using biochemical testing. Using capsular multiplex PCR and multilocus sequence typing, serotypes and genotypes of the clinical strains of P. multocida were identified. Results Capsular multiplex PCR revealed capsular serotypes A and B among the 14 clinical strains of P. multocida, which comprised 71. 4% ( 10 / 14) and 28. 6% ( 4 / 14), respectively. Six sequence types (STs) were detected by multilocus sequence typing: ST12, ST35, ST72 and ST73, and two new types, ST76 and ST77, described for the first time here. Conclusions Genotype diversity was observed in P. multocida isolated from laboratory rabbits, and the capsular serotype A, which is associated with respiratory disease, accounted for a relatively high proportion of isolates. This study reinforces the importance of quality control for rabbit breeding.

Abstract:Objective To investigate the regulatory effect of resveratrol ( RSV) targeting of SIRT1 ( silence information regulator) to activate the kelch-like ECH associated protein (Keap) 1-nuclear factor erythroid 2-related factor (Nrf)2-heme oxygenase (Ho)-1 pathway on calcium oxalate stone formation. Methods In this experiment, the optimal drug dose of RSV for human renal epithelial cells (HK-2) was screened by the Cell Counting Kit (CCK)8 method . HK-2 cells were randomly divided into four groups: control, oxalic acid, oxalic acid + RSV, and oxalic acid + RSV+ EX527. The cell viability of the preparation group was determined by the CCK8 method . Flow cytometry was used to detect the reactive oxygen species (ROS) production in each experimental group. The expression of Sirt1, Keapl, Nrf2, HO-1, transforming growth factor (TGF)-β1 and Smad2 were explored by western blotting and PCR experiments. A HK-2-small interfering RNA (siRNA) was constructed, and the above indicators were re-tested. Results Drug toxicity screening result showed that 500 nM was the optimal drug dose for the experiment. CCK8 suggested that RSV relieved the damage of HK-2 by oxalic acid. The Sirt1 inhibitor EX527 reversed the protective effect of RSV. The measurement of active oxygen levels showed that RSV reduced the production of active oxygen induced by sodium oxalate, and EX527 blocked the effect of RSV. Western blotting result showed that, compared with the sodium oxalate group, Sirt1, Keap1, Nrf2 and HO-1 expression was upregulated (P < 0. 01) while Smad2 expression was downregulated ( P < 0. 01). However, after applying EX527, the expression of related proteins upregulated by Sirt1 decreased. Quantitative PCR result showed that, compared with the sodium oxalate group, mRNA expression of TGF-β1 (P<0. 01), Smad2 (P<0. 05), Smad3 (P<0. 05) and Smad4 (P< 0. 01) were downregulated (P<0. 01) after RSV application, while Sirt1 mRNA expression was upregulated (P<0. 01). The application of EX527 reversed these effects. Western blotting result showed that, compared with the sodium oxalate group, Sirt1 expression was downregulated, and keap1, Nrf2 and HO-1 expression were downregulated after siRNA interference. When RSV treatment was applied after silencing of Sirt1 expression, RSV could not inhibit Smad2 activation. Conclusions This study showed that RSV had antioxidant effects on HK-2 cells through a mechanism that may involve Sirt1 activation to promote the expression of Keap1, Nrf2 and Ho-1, which further inhibits the production of TGF-β1, thereby inhibiting the generation of ROS.

Abstract:Objective To investigate the protective effects of curdione on cognitive and neurological function in mice with cerebral ischemia-reperfusion injury. Methods A middle cerebral artery occlusion mouse model was developed using middle cerebral artery embolization ( equivalent normal saline; model group; 20). Mice received the following treatments: low-molecular-weight heparin sodium [1 mg / ( kg·d); HS group; 20], low-dose curdione [20 mg / ( kg·d); CD-L group; 20] or high-dose curdione [ 60 mg / ( kg·d); CD-H group; 20]. We observed changes in behavioral evaluation, Morris water maze, cerebral infarction volume and cerebral water content, and measured the content of 6-keto prostaglandin F1α (6-keto PGF1α), thromboxane B2 (TXB2), 6-keto-PGF1α/ TXB2 in serum, and cAMP and p-CREB in brain homogenates. Results Compared with the sham operation group, the escape latency in the model group was significantly increased. Number of passes through the platform was obviously reduced (P< 0. 01). Behavioral evaluation, cerebral infarction volume, and content of cerebral water(%)and TXB2(pg / mL)were obviously increased. The content of 6-keto-PGF1α, 6-keto-PGF1α/ TXB2 ratios, cAMP and p-CREB were all significantly decreased (P< 0. 01). Compared with the model group, the escape latency in the CD-L and CD-H group was significantly decreased. The numbers of passing through the platform were obviously increased (P<0. 05). Behavioral evaluation, cerebral infarction volume, the content of cerebral water and TXB2 were obviously decreased. The content of 6-keto-PGF1α, 6-keto-PGF1α/ TXB2 ratios, cAMP and p-CREB were significantly decreased (P<0. 05), and appeared dose-dependent. Conclusions Curdione had a protective effect on cognitive function and neurological function in mice with cerebral ischemia-reperfusion injury. Its mechanism may be related to the reduction of TXB2, the increment of 6-keto-PGF1α, 6-keto-PGF1α/ TXB2 ratio, cAMP and p-CREB, thereby improving microcirculation disorders in cerebral ischemia reperfusion injury in a manner involving activation of the cAMP / CREB/ BDNF signaling pathway.

Abstract:Objective To explore the role and mechanism of inhibitor of differentiation 1 (Id1) on replication of the hepatitis B virus, and proliferation and apoptosis of hepatoma cells. Methods The Id1 gene in the HepG2. 2. 15 hepatoma cell line was silenced by RNA interference (siRNA-Id1 group). An unrelated sequence was designed and used for the siRNA-Control group. The expression of p53 in siRNA-Id1 cells was inhibited by pifithrin-α ( siRNA-Id1+Pifithrin-α group) and HepG2. 2. 15 hepatoma cells were used as the control group. The replication of hepatitis B virus, cell proliferation, and apoptosis were detected by qRT-PCR, western blot, ELISA, MTT, and flow cytometry, respectively. Results The mRNA and protein expressions of Id1 in the siRNA-Id1 group were lower than those in the siRNA-Control group (P<0. 001). Compared with the siRNA-control group, the mRNA and protein expressions of HBx in the siRNA-Id1 group were decreased (P<0. 001), the contents of HBsAg and HBeAg in the supernatant were decreased (P<0.001), the proliferation rate of cells was decreased (P<0. 05), the proportions of early apoptosis cells and late apoptosis cells were increased (P<0. 001), the protein expressions of Bax and cleaved caspase-3 in cells were increased (P<0. 001), and the protein expression of BcL-2 was decreased (P<0. 05). Pifithrin-α partly inhibited these biological effects induced by Id1 gene silencing. Conclusions The RNA interference of Id1 reduced the replication of hepatitis B virus, inhibited the proliferation of hepatocellular carcinoma cells, and induced cell apoptosis. The mechanism may be related to the overexpression of p53.

Abstract:Objective To investigate the effects of different types of exercise on emotion and behavior in aggressive rats. Methods Fifty-six 12-week-old SPF male SD rats were randomly divided into the following five groups after 1 week of adaptive feeding: quiet ( A), aggressive model ( G), aggressive jumping ( GT), aggressive treadmill (GP), and aggressive (R). Rats were fed in single cages, subjected to invasion by foreign rat to establish the aggressive model, and observed for changes in emotional and behavior. Interventions with jumping and treadmill exercises for 8 weeks the aggressive jumping were training for 3 days a week, and the treadmill were training for 5 days a week. in the middle and late stage of training, the open-field experiments were conducted to observe changes in emotion and behavior after exercise intervention. Results The exploratory behaviors, degrees of activity and autonomous activity of the rats in the aggressive model (G) group were significantly reduced, the anxious mood and tension were higher. After 4 weeks of jumping and treadmill exercise intervention, there were significant differences in the number of upright times, crossing numbers, washing and shaving times between the aggressive jumping (GT) and the aggressive treadmill (GP) groups. The average speed, central activity time and total distance of the two groups of rats were significantly increased. The excitability and exploration effects of the two kinds of exercises on the aggressive model (G) group were significant, and the influence of the jumping exercise on rat behavior was more obvious. After 8 weeks of jumping and treadmill exercise intervention, the excitability of rats in the aggressive jumping (GT) and aggressive treadmill (GP) groups increased significantly, but the excitability, exploratory behavior and spontaneous activity of rats following the treadmill exercise intervention decreased significantly, and there was no significant difference with the aggressive model (G) group. Conclusions Proper jumping and treadmill exercise significantly improved the anxiety, nervousness and proactive behavior of the aggressive rats.

Abstract:Objective To investigate the protective effect and mechanism of activated protein C ( APC) on myocardial ischemia reperfusion injury in rats. Methods Thirty male SD rats were randomly divided into sham operation group (10), I/ R group (10) and apc-treated group (10). The anterior descending branch of the left coronary artery was ligated for 60 min, and the myocardial ischemia reperfusion model was established 6 h after reperfusion. The expression levels of TNF-α, MPO and ICM-1 were detected by ELISA and immunohistochemistry. HE staining was used to detect the changes of myocardial pathology in each group.Results Compared with the sham group, the expression of TNF-α, MPO and ICAM-1 were increased significantly in the I/ R group (P< 0. 01); but compared with the I/ R group, the expression of TNF-α, MPO and ICAM-1 were obviously decreased in the APC group (P< 0. 01). Conclusions APC has a protective effect on myocardial I/ R injury, in a mechanism that may involve inhibiting the expression of inflammatory factors and reducing the infiltration of inflammatory cells.

Abstract:Intrauterine growth retardation ( IUGR) can cause persistent impaired development of the pancreas in fetuses. Meanwhile, IUGR can cause dyspepsia and disorganized glucose metabolism, increasing the risk of metabolic diseases in offspring and even leading to the premature death of animals. As a secretory organ, impairments in internal and external secretory functions of the pancreas are to a large extent due to the improper development of the pancreas. This paper reviews the regulatory mechanisms of IUGR that affect transcription factors important at various stages of pancreas development, facilitating research to clarify the pathophysiological mechanisms associated with IUGR and impaired pancreas development. Ultimately such research will lead to new, reasonable and effective measures to prevent and treat IUGR.

Abstract:Brucellosis remains one of the most common zoonosis globally, affecting both animal and human health. In China, the number of human brucellosis cases continues to grow due to the lack of an effective licensed vaccine. After infection by Brucella, the bacteria can adapt to the host macrophage internal environment through regulating the expression of its own genes, ensuring its resting survival and subsequent replication, which hinders effective therapeutics for brucellosis. This review focuses on the issues of Brucella species and its host range, animal infection models and vaccine investigation, summarizes the mechanisms of Brucella intracellular survival within the host, and proposes key points for the prevention and control of brucellosis.

Abstract:Premature ovarian insufficiency ( POI) is a clinical syndrome characterized by menstrual disorders accompanied by high gonadotropin and low estrogen in women under the age of 40. POI causes destructive damage to a woman’s endocrine function, fertility and even normal life. Understanding the signal transduction pathways related to POI is of great significance in explaining the causes of POI, restoring ovarian function and finding potential drug treatment targets and new therapeutic technologies. In this review, we summarize the recent research advances of POI-related signaling pathways, mainly focusing on the transforming growth factor-β ( TGF-β) superfamily, PI3K-AKT and Hippo signaling pathways.

Abstract:Allergic diseases have become a global public health problem that not only affects the quality of life of patients, but can also be life threatening. Using mouse models, a large number of molecular mechanisms and conformational changes of various allergic diseases have been revealed, such as allergic conjunctivitis, allergic asthma, contact dermatitis and food allergy. Although the types and phenotypes of immune responses in allergic mice with different diseases vary, they are similar to the immune responses and phenotypic changes that occur in humans. In this review, we summarize the common method for establishing mouse models of allergic diseases and describing the phenotypic changes of each. We hope that this review will provide a reference for the continued establishment and quality evaluation of allergic animal models in mice.

Abstract:Polycystic ovary syndrome ( PCOS) is a gynecological metabolic and endocrine disorder of unknown etiology. Exploring different PCOS animal models might help reveal the etiology of PCOS and establish prevention and treatment measures. In this paper, we searched China National Knowledge Infrastructure (CNKI) for “ polycystic ovary syndrome + animal model” and PubMed for “ polycystic ovary syndrome + animal model ”, excluding reviews and theoretical literature. We found 231 articles from 1989 to 2018, and PCOS animals included research literature on rodent, non-human primate, and ruminant models. This study evaluated similarities between different animal models and human PCOS, to indicate the optimal model for different research purposes of PCOS disease research.

Abstract:Wild-type p53 induced phosphatase 1 ( Wip1) is encoded by Ppm1d, a member of the protein phosphatase type 2C ( PP2C) phosphatase family and has an important role in several stress signaling pathways. Many previous studies have shown that Wip1 has a role in regulating transcription factors as well as being a p53 target. For example, Wip1 is crucial for cellular homeostasis by creating a negative feedback loop with p53, p38 / MAPK, and other factors. Thus, Wip1 plays a vital role in the regulation of the cell cycle and inhibiting apoptosis, DNA repair, and inflammation. In this review, we will discuss the function Wip1 related to these functions.