Abstract:Objective　To investigate the effect of astragaloside IV　in combination with ferulic acid on the proliferation of cobalt chloride-induced hypoxic human umbilical vein endothelial cells (HUVECs) and its mechanism.Methods　A hypoxic injury model of HUVECs was induced by cobalt chloride. CCK-8 and living cell staining techniques were used to detect cell proliferation, fluorescence staining was used to observe the morphological changes of cells, a scratch test was used to determine the migration ability of the cells, and a tube test was used to observe the effect of drugs on endothelial cells. Western blot was used to detect the expression of p-JAK2 and p-STAT3 proteins. Results　Compared with the model group, the combination of astragaloside IV and ferulic acid significantly promoted the proliferation of HUVECs induced by cobalt chloride for 24 hours, improved morphological changes in cells induced by hypoxia, and promoted cell migration and angiogenesis. Concurrently, the protein expressions of p-STAT3 and p-JAK2 were upregulated.Conclusions　Astragaloside IV combined with ferulic acid protects against cobalt chloride-induced hypoxia in HUVECs and promotes angiogenesis. The mechanism may be related to activating the JAK-STAT signaling pathway.

Abstract:Objective 　To detect and analyze the expressions of serum inflammatory factors in chronic renal disease (CKD) rats with vascular calcification by antibody microarrays. Methods　Thirty SD male rats were randomly divided into control and model groups. The model group was treated with adenine gavage and adriamycin caudal intravenous injection to establish a model of CKD with vascular calcification, while the control group was given corresponding saline treatment. Twenty-eight days after administration, the levels of serum creatinine, urea nitrogen, blood calcium and blood phosphorus were measured. Sirius red staining, HE and von Kossa staining were used to detect morphological changes in renal and aortic tissues. Immunohistochemical staining was used to detect the expressions and distributions of α-smooth muscle actin (α-SMA) and Runt related transcription factor 2 (Runx2) in the aorta. Antibody microarray was used to detect 27 inflammatory cytokines in serum samples from the two groups. Results　Compared with the control group, serum creatinine, urea nitrogen, calcium, and phosphorus levels were increased. Sirius red staining showed marked renal fibrosis,and HE and von Kossa staining showed elastic fiber rupture and calcium deposition in aortic media. Immunohistochemical staining showed that the expression of α-SMA was decreased and that of Runx2 was increased in the model group compared with the control group. Antibody microarray results showed that compared with the control group, the levels of CINC, GMCSF,IFN-γ, IL-1α, IL-6, MCP-1, ICAM-1, TIMP-1, L-selectin, IL-2 and IL-10 were increased by different degrees from those in the control group, while IL-4, IL-13, fractalkine, IL-1β and TNF-α were lower than in the control group.Conclusions　There is a significant imbalance in the levels of inflammatory factors in CKD rats with vascular calcification,in which proinflammatory factors play a leading role.

Abstract:Objective　To study the physical fatigue behavioral manifestation of mice after sleep interruption (SI)at different timepoints, to provide a stable and reliable animal model for the research and development of general health products. Methods　Mice were placed into a rotating drum sleep interferometer for different periods of time (5, 10, and 15 days). Then, the rotating wheel and loaded swimming tests were used to assess the behavioral activities of mice after SI.The hepatic glycogen and serum urea nitrogen levels were used to assess the biochemical index after SI. Results　In the rotating wheel exhausted test, the exhausted time of SI in the 5 d, 10 d and 15 d groups were decreased compared with the control group, but with a non-significant difference ( P > 0. 05). In the loaded swimming test, compared with the control group, the mice after SI for 10 and 15 days showed significantly decreased exhaustive time ( P < 0. 05, P < 0. 01), sink frequency and total sink time ( P < 0. 01, P < 0. 01). In addition, our study demonstrated that the content of hepatic glycogen was significantly reduced in mice after SI for 5, 10, and 15 days ( P < 0. 05) compared with the control group,and that the serum urea nitrogen level of mice after SI for 10 and 15 days was increased ( P < 0. 05). Conclusions　A mouse model of physical fatigue behavior can be established by sleep interruption using a sleep interruption apparatus set at rotation speed, 20 s per revolution; and rotation frequency, rotating 1 min after 1 min pause for 10 days and 15 days.

Abstract:Objective　To observe the effects of Wenjingzhitong decoction (WJZT) on expression of molecules in the AKT/ mTOR pathway secreted by eutopic and ectopic endometria of endometriosis (EMS) rats. Methods　Autologous transplantation of the uterine was used to establish the rat model of endometriosis. Seventy-five endometriosis rats were randomly divided into five groups: WJZT high-, medium-, and low-dose groups, and Nemestran and model control groups.Sham surgery rats were used for the normal control group. The WJZT high-, medium-, and low-dose groups were treated with different concentrations of Wenjingzhitong decoction, respectively. Nemestran (gestrinone) capsule was administered to the Nemestran group, and 0. 9% NaCl was administered to model and normal groups. AKT and mTOR mRNA and protein expressions were detected by real-time PCR and immunohistochemistry, respectively. Serum levels of vascular endothelial growth factor (VEGF) were detected by ELISA. Results　Compared with the normal control group, protein and mRNA expressions of AKT and mTOR in eutopic and ectopic endometria was increased in the model groups ( P < 0. 01). AKT and mTOR mRNA levels in eutopic and ectopic endometria of the WJZT high-dose group and Nemestran (gestrinone) group was lower than that in the model control group ( P < 0. 05). Compared with the normal control group, serum VEGF level in the EMS model groups was increased ( P < 0. 01). Compared with the model control group, serum VEGF levels in the WJZT high- and low- dose groups and the Nemestran group were decreased ( P < 0. 01), and serum VEGF level in the WJZT medium-dose group was also decreased ( P < 0. 05). Conclusions 　Endometriosis is associated with the AKT/ mTOR signaling pathway and the levels of VEGF. WJZT inhibits AKT/ mTOR and regulates the level of VEGF. Therefore, WJZT can inhibit cell adhesion, invasion, and angiogenesis in the ectopic endometrium.

Abstract:Objective 　To establish a model of oral mucositis in golden hamsters induced by 5-fluorouracil combined with mechanical scratching. Methods　Male golden hamsters were divided into a control group and model group.Animals in the model group were intraperitoneally injected with 60 mg/ kg and 40 mg/ kg of 5-fluorouracil (5-FU) on the first and second days, and the right cheek pouch was slightly scratched by an 18-gauge needle on the fourth day. The control animals were intraperitoneally injected with an equal volume of normal saline on the first and second days. After the treatment, the pathological changes of the right cheek pouch of each animal were scored every day and the body weight and the mean daily food intake of each animal were recorded. The level of proinflammatory cytokines TNF-α and IL-1β in the serum was measured by ELISA. The right cheek pouch tissue was removed for pathological examination on days 8 and 14 after the treatment, and the contents of malondialdehyde (MDA) and superoxide dismutase (SOD) activity in the right cheek pouch tissue were determined by biochemical analysis. Results　The model group developed an ulcer on the surface of the right cheek pouch on the third day after treatment, and the macroscopic scores of right cheek pouch were the highest compared with the control group. The mean daily food intake was significantly reduced and the body weight was decreased within 8 days after the treatment in the model group. On days 8 and 14 after modeling, the histological structures of the right cheek pouch tissues were damaged and had an inflammatory reaction. The levels of TNF-α and IL-1β in the serum of golden hamsters were significantly increased, the MDA content in the cheek pouch tissue was significantly increased, and the SOD activity was reduced on days 8 and 14. Conclusions 　5-fluorouracil combined with mechanical scratching induces the formation of oral mucositis in golden hamsters. The inflammatory response is marked within 8 days after the treatment, and the most obvious is on the third day. Therefore, this model can be used to study the pathogenesis of oral mucositis and for the development of therapeutic drugs.

Abstract:Objective　To investigate the mechanism of aspirin on high salt-induced hypertension in Dahl saltsensitive rats. Methods　Two-month-old Dahl salt-sensitive rats (Dahl SS) and salt-resistant rats (SS-13BN) were fed with Low salt (0. 12% NaCl, LS), High salt (8% NaCl, HS), High salt + Aspirin (8% NaCl +10 mg/ (kg·d)aspirin),or HS+ASA for 8 weeks. Blood pressure was measured by the tail cuff method. The expressions of renal inflammatory cytokines (IL-6, IL-1β, TNF-α) and eNOS and vWF in arteries were measured by real-time PCR and western blot,respectively. The number of skin M2 macrophages was measured by immunofluorescence. Results 　Blood pressure,expressions of renal IL-6, IL-1β, TNF-α, and arterial vWF were markedly increased, and the number of M2 macrophages in the skin, and eNOS expression in the arteries were significantly decreased in the high salt group compared with the low salt group. Aspirin inhibited all these events in rats on a high salt diet. However, the influence of high salt and aspirin was not observed in SS-13BN rats. Conclusions　Aspirin attenuated salt-sensitive hypertension and arterial injury induced by a high salt diet in Dahl salt-sensitive rats via inhibiting platelet activated inflammation.

Abstract:Objective 　To breed and identify FOXO 3A gene knockout mice, hybrid mice were bred for conservation and homozygous mice were used for experiments. The effect of FOXO 3A gene knockout on the hematopoietic cell phenotype in mice was preliminarily analyzed, laying the foundation for subsequent studies on the regulation of the FOXO 3A gene during hematopoietic cell injury. Methods 　Hybrid female mice were mated with wild type male mice.Female and male hybrid mice were selected and bred, and homozygous mice were bred using one male and one or two females per cage. Pedicle marking and tail snips of neonatal mice were used to extract genomic DNA, and PCR was used for genotype identification. Heterozygous mice ( FOXO 3A+/ - ) with 100/186 bp bands, homozygous mice ( FOXO 3A-/ - ) with 186 bp bands, and wild type mice ( FOXO 3A+/ + , WT) with 100 bp bands were identified. Wild type and homozygous mice were used for bone marrow cell typing by flow cytometry. Results　 FOXO 3A gene knockout mice were successfully bred and identified. Homozygous and heterozygous mice from the daughters of gene knockout mice had normal growth, with no obvious difference in appearance or development compared with wild type FVB/ N mice. A number of homozygous mice were obtained for subsequent experiments. The bone marrow cell count of FOXO 3A knockout homozygous mice contained similar numbers of nucleated cells compared with wild type mice (6. 167±1. 424 vs 10±1. 732; P = 0. 1625). The results of flow analysis showed that the proportion of hematopoietic progenitor cells (Lin-Scal-1- c-kit+ , HPC) in the bone marrow of FOXO 3A knockout mice was significantly increased ( P <0. 05) compared with the wild type mice. In addition, there was no significant difference in the proportion of hematopoietic stem cells (Lin-Scal-1+ c-kit+ , HSC), or number of hematopoietic stem cells and hematopoietic progenitor cells ( P >0. 05). Conclusions　 FOXO 3A gene knockout mice were successfully obtained by genotype identification. Our preliminarily study demonstrated that FOXO 3A gene knockout had no significant effect on the bone marrow cell count or typing in mice. Future studies will investigate the effect of FOXO 3A gene knockout on hematopoietic cells in mice.

Abstract:Objective 　To explore the expression and mechanism of BDNF and TrkB receptors under acute hypoxia and hypoxic preconditioning, and to provide a reference for the study and clinical application of hypoxic preconditioning. Methods　A model of acute hypoxia and hypoxic preconditioning was generated in ICR mice. After 0-4days, the hippocampus was isolated from the brains of hypoxic mice, and the protein and gene expressions of BDNF and its receptor TrkB were detected by Western blot and real-time PCR. Results　The study found that the tolerance time was increased significantly with the increased amount of hypoxia in mice ( P <0. 05). Compared with the control group, the expression of BDNF and the full-length TrkB receptor in the hypoxia group was increased, and the expression of BDNF protein was significantly increased in the early phase of hypoxic preconditioning ( P <0. 05). Compared with the control group, the expression of the truncated TrkB receptor was decreased, and the expression of mRNA was significantly decreased in the middle and late phase of hypoxic preconditioning ( P <0. 05). Compared with the control group, the activity of the BDNF/ TrkB signaling pathway was inhibited in the acute hypoxia group and increased in the hypoxic preconditioning group. The activity of the BDNF/ TrkB signaling pathway was significantly increased in the late phase ( P < 0. 05).Conclusions 　Hypoxic preconditioning may be mediated by upregulating the binding between BDNF and TrkB,downregulating the expression of truncated TrkB, reducing the formation of heterodimers, and co-activating the BDNF/ TrkB signaling pathway, which ultimately has a neuroprotective effect in mice.

Abstract:Objective　To investigate the inhibitory effect of Shenyi capsules combined with 5-FU on the growth and angiogenesis of colon cancer and its possible mechanism. Methods　Fifty BALB/ c mice with LoVo colon cancer were randomly divided into five groups: model group, 5-FU group, Shenyi capsule low-dose group, Shenyi capsule high-dose group, and Shenyi capsule+5-FU group. Differences in disease indicators were compared. Results　1) The Shenyi capsule high-dose and Shenyi capsule+5-Fu groups had the highest inhibition rate, and with the extension of administration time,the inhibition was more marked. (2) The thymus mass, thymus index, spleen mass and spleen index of the Shenyi capsule +5-FU group were improved significantly ( P <0. 05). (3) The inhibitory effect in the Shenyi capsule+5-FU group on MVD was the most obvious, followed by the Shenyi capsule high-dose group and low-dose group. An inhibitory effect of Shenyi capsule+5-FU group on the MVD of tumor was not observed. (4) Compared with the model group, the metastasis rates in various organs in the high dose Shenyi capsule group and Shenyi capsule+5-FU group were significantly lower ( P <0. 05).(5) Compared with the model group, the expressions of Gli1 protein and VEGF protein were decreased significantly ( P <0. 05) in the Shenyi capsule high-dose group and Shenyi capsule+5-FU group. Conclusions　Shenyi capsule inhibits the growth and angiogenesis of colon cancer cells, and VEGF may be its main target.

Abstract:Objective　To detect and compare the clinical outcomes of multiple serum cytokines in healthy Rhesus and Cynomolgus monkeys. Methods　Fifteen healthy Rhesus and 15 healthy Cynomolgus monkeys were chosen for sample collection. Serum concentrations of 43 cytokines were measured using xMap technologies. The results were analyzed by t-test or non-parametric Mann-Whitney U-test according to the W-test for normality. Results　Among 43 cytokines, one proinflammatory cytokine (IL-15), three anti-inflammatory cytokines (IL-1RA, IL-6R α, and sCD27) and one chemokine (CCL5/ RANTES) were significantly different between Rhesus and Cynomolgus monkeys. Conclusions 　There is no significant difference in basic serum cytokine levels between Rhesus and Cynomolgus monkeys, which is comparable in biomedical research. The results of this study provide basic data for related studies.

Abstract:Objective 　To observe the effect of Maca combined exercise therapy on mitochondrial respiratory enzyme activity and renal function in diabetic rats. Methods　Forty diabetic rats were randomly divided into model control group, Maca group, training group and Maca + training group. The training and Maca + training groups underwent treadmill exercise training every day (5 g/ kg). After 6 weeks, rats in each group were subjected to exhaustive exercise (35 m/ min)followed by immediate extraction of renal mitochondria under anesthesia. Activity of the mitochondrial respiratory chain enzymes RCCI, RCCII, RCCIII and RCCIV, blood urea nitrogen (BUN), serum creatinine (SCr), and blood glucose contents were measured. Results　Compared with the control group, the Maca group RCCII, training group RCCI and RCCII, and Maca + training group RCCI, RCCII and RCCIV activities were significantly increased. Compared with the Maca group, RCCI and RCCII activities in the Maca + training group were significantly increased. Compared with the training group, the RCCII activity in the Maca + training group was significantly increased. Compared with the control group, the content of blood urea nitrogen in the Maca and Maca + training groups, and the levels of serum creatinine and blood glucose in the training group and Maca + training group were significantly decreased ( P < 0. 05 or P < 0. 01).Compared with the Maca group, the levels of serum creatinine and blood glucose were significantly decreased in the Maca +training group ( P < 0. 05). Compared with the training group, the levels of blood urea nitrogen and blood glucose in the Maca + training group were significantly decreased ( P < 0. 05). Conclusions　Maca combined with exercise therapy improved the respiratory function, increased the level of oxidative phosphorylation, and improved kidney function in aged diabetic rats.

Abstract:Objective　To investigate the protective effect of dexmedetomidine to regulate TLR4 expression on oxygen glucose deprivation injury in neurons and its mechanism. Methods　PC12 cells were cultured in vitro, and the oxygen glucose deprivation cell model was generated. The cell survival rate was detected by CCK-8 test, the expressions of Bax, Bcl-2 and TLR4 proteins were detected by Western blot, the expression of TLR4 mRNA was detected by RT-PCR,and the levels of TNF-α and IL-6 were detected by ELISA after treatment with low (0. 1 μmol/ L), medium (1. 0 μmol/ L)and high doses (10. 0 μmol/ L) of dexmedetomidine for 24 h. In the replicating oxygen-deprived cell model, the expression of TLR4 was inhibited by the TLR4 inhibitor TAK-242. Dexmedetomidine altered the cell survival rate, and levels of Bax,Bcl-2, TLR4, TNF-α and IL-6 proteins. Results　The survival rate of PC12 cells was increased, the expressions of Bax,TLR4, TNF-α, and IL-6 proteins, and TLR4 mRNA were decreased, and the expression of Bcl-2 protein was increased ( P <0. 05). This was concentration-dependent after treatment with low, moderate, and high dose dexmedetomidine. After TAK-242 treatment, the expression of TLR4 was decreased, the survival rate of PC12 cells was increased, the expressions of Bax, TNF-α, and IL-6 proteins were decreased, while the expression of Bcl-2 protein was increased ( P <0. 05). The effect of TAK-242 was enhanced after treatment with dexmedetomidine ( P < 0. 05). Conclusions 　Dexmedetomidine protected neurons from oxygen glucose deprivation by inhibiting the expression of TLR4.

Abstract:Objective　To explore the effects of exposure to perfluorooctane sulphonate (PFOS) at different doses on embryonic development and expression of placental insulin-like growth factor 1 ( IGF-1) in rats. Methods 　At gestational day 12, pregnant rats were exposed to different doses of PFOS (0, 5, 10, and 20 mg/ kg) by gavage for 7 d. At gestational day 19, the body masses of pregnant and fetal rats, liver organ coefficient of fetal rats, serum PFOS and glucocorticoid (GC) levels of pregnant rats, and IGF-1 mRNA level of placenta were measured. Results　Compared with those in the control group, the serum PFOS levels of pregnant rats in the treatment groups were significantly higher with increasing PFOS dose, while the body mass of pregnant rats and the body mass and length of fetal rats exposed to 20 mg/ kg PFOS were significantly lower ( P < 0. 001). Fetal rat liver organ coefficient in the treatment group showed the same trend of a marked decrease ( P < 0. 05). Blood biochemical alanine aminotransferase (ALT) and aspartate aminotransferase (AST) also increased with the dose of PFOS ( P < 0. 05). Treatment with 10 mg/ kg PFOS increased the serum GC level of pregnant rats ( P < 0. 05). IGF-1 mRNA level decreased with the increasing PFOS dose in placenta. Conclusions　PFOS exposure during pregnancy can lead to liver toxicity in fetal rats through the maternal accumulation of PFOS in the serum,increased GC concentration, and decreased IGF-1 mRNA expression, thereby affecting the growth and development of offsprings.

Abstract:Objective　To detect 25% cyhalofop-butyl emulsifiable concentrate(EC) skin sensitization by a local lymph node assay (BrdU-ELISA) in mice. Methods　Female BALB/ C mice were tested and the samples were set up to 100%, 50%, and 25%. The negative control group received acetone: olive oil (AOO)= 4 ∶1, and the positive control group received 25% basal cinnamaldehyde. Test samples were laid on the dorsum of bilateral mouse ears for three consecutive days, then the mice received an intraperitoneal injection of BrdU (5 mg/ mouse) on the sixth day. After 24 hours, the lymph nodes were removed, weighed, and mashed into a cell suspension. Lymphocyte proliferation was measured by BrdU-ELISA. Results　The body weights of female mice in all treated groups were similar to the control group. The increase of ear thickness and ear weight in each dose group was not more than 25%. The stimulus indexes of solvent and blank control groups were SI₁and SI₂< 1. 6, and the stimulation indexes of the positive control group (25% hexyl cinnamaldehyde) were SI₁and SI₂ > 1. 6. This indicated the test system was reliable. The stimulation indexes of 25%cyhalofop-butyl EC in the low, medium and high dose groups were SI₁and SI₂ < 1. 6, and there was no dose-response relationship. Conclusions 　The results of 25% cyhalofop-butyl EC in mouse local lymph node assays were negative,indicating that 25% cyhalofop-butyl does not cause skin sensitization.

Abstract:Objective　To verify the reproducibility of microsatellite detection for the genetic quality control of laboratory animals in three different laboratories. Methods 　DNA of six laboratory mice was amplified using 12 microsatellites in three laboratories. PCR products were analyzed by electrophoresis and sequencing. Results 　Clear electrophoresis bands and 72 alleles were obtained in the Beijing laboratory of National Institutes for Food and Drug Control (NIFDC), the Zhejiang laboratory, and the Guangdong laboratory. Sequencing results showed that the alleles identified were the same in the three different laboratories. Conclusions　Microsatellites can be used for the genetic quality control of laboratory animals, with a good reproducibility between different laboratories.

Abstract:近些年大量的文献对实验大小鼠采血方法进行了比较研究,以探究出最适合的采血方法,从而获得高质量的血液样本。本文主要介绍了几种实验大小鼠常用的采血方法,及不同采血方法对动物福利的影响,为选择最适的采血方法提供相关的理论依据,提高实验效率,降低实验影响。

Abstract:Objective　To summarize the characteristics of two main individually ventilated cage (IVC) air supply modes in the construction of laboratory animal facilities, and analyze their different uses. Methods　According to typical cases, the specific characteristics of the two air supply modes were analyzed in detail by five aspects: safety and reliability,space utilization, operation energy consumption, monitoring and regulation and initial investment. Results 　For each comparison item, the advantages and disadvantages of the two air supply modes were compared. Conclusions　The two method of air supply had their individual advantages. Air supply mode A is suitable for small-scale facilities and projects that require a decentralized and flexible control of cage parameters. Air supply mode B is suitable for medium-sized and large-scale facilities.

Abstract:Hand, foot and mouth disease (HFMD), which has spread worldwide, is a common childhood infectious disease and a serious threat to children’s health. HFMD is caused by more than 20 enteroviruses, especially enterovirus 71 (EV71). Many studies have shown that HFMD results from the combined effects of virus, host, and environment. After virus invasion, various pathological reactions are caused by escape from surveillance of the host’s natural or acquired immune surveillance. This is not only related to the infection characteristics of the virus, but also to host-related genes such as receptor genes and immune response genes. In recent years, pathogenic host genes related to HFMD have become hot spots for researchers. However, some of the pathogenic related host genes are closely related to regions, races, or populations, and the research conclusion regarding HFMD in different populations in different regions are often not consistent. Therefore, it is necessary to find widely applicable host genes that are closely related to HFMD.Genetic diversity mice are a new tool for simulating population genetic diversity and studying complex traits or diseases. In this paper, we review research on HFMD pathogenicity and related host genes, as well as the advantages of genetic diversity mice in HFMD research.

Abstract:To select the appropriate mouse model of liver injury is of great significance for the research on prevention and treatment of liver disease, the study of its pathogenesis and the screening of drugs to protect the liver.According to references published internationally, mouse models of liver injury were classified into chemical,pharmacological, immunological, alcoholic, and partial hepatectomy studies. Based on the classification of models of liver injury in mice, the pathogenesis, dosage of drugs, time of model formation, applicable scope, and advantages and disadvantages were reviewed to provide a reference for the selection of experimental mouse liver injury models.

Abstract:Nervous system diseases in the central nervous system, peripheral nervous system and autonomic nervous system lead to sensory, motor, consciousness, and autonomic dysfunctions, which are the main manifestations of these diseases. They have a marked impact on a patient’s life and work. In traditional medicine, cinnamon has the function of “ nourishing the primary yang, warming the spleen and stomach, removing internal cold, and promoting blood circulation.” Modern research suggests that cinnamon has anti-oxidation, free radical scavenging, anti-inflammation, antiintermittency,and anti-ulcer effects. Recent studies have shown that cinnamon has a medicinal effect on nervous system diseases, such as Alzheimer’s disease, Parkinson’s disease and multiple sclerosis. Therefore, this paper summarizes the research progress and potential medicinal value of cinnamon for the treatment of nervous system diseases.