Abstract:Objective Construction of a type 2 diabetic (T2DM) model in rhesus monkey, so that to become the effective substitute for the study of human T2DM. Methods Based high glucose and fat diet, to appearance hyperlipidemia and obesity status followed by injection of 35 mg/kg of streptozotocin (STZ). The body mass index(BMI), lipid profile, fasting blood glucose, insulin, insulin resistance index, urine sugar and oral glucose tolerance test were measured, and some of its clinical features were analyzed. Results The BMI greater than 35 mg/kg reach to severe obesity shown a characteristics of hyperlipidemia, blood glucose, insulin and insulin resistance index were significantly elevated (P <0.01), urine sugar test were positive, and glucose tolerance were impaired, and fasting glucose of higher than 7 mmol/L and glucose levels higher than 11.1mmol/L in two hours in the T2DM model group. Conclusion Analysis some of the clinical features, the model group had the typical T2DM clinical features. So it will be an effective model in human T2DM research.

Abstract:Objective To compare the effect of the two vitrification of embryos frozen invarious mouse strains . Methods The two-cell embryos from six mouse strains(KM、ICR、BALB/c、C57BL/6J、OB/OB、LAP/rtTA0F59) were cryopreserved and thawed respectively with EFS and DAP cryoprotectant . The thawed rate in vitro and the implantation rate in vivo were determined to evaluated the two ways. Results The postthaw survival rates of different strains in EFS were average 69.97% ( 47.9%～83.6% ), and DAP were average 47.23%(26.3%～76.7%). After 2-cell embryos were transferred to the oviducts of pseudopregnant recipient in all strains, the implantation rates in EFS were ranged from 1.75% to 45.0% (average 27.23%) and DAP were ranged from 7.0%～46.3% (average 31.43%). With the respect of thawed rates, EFS was preferred to DAP. No significant differences(P＞0.05) were found in implantation rates of EFS and DAP on KM and ICR strains. But the implantation rates of BALB/c、C57BL/6J、OB/OB、LAP/rtTA0F59 in EFS were higher than those in DAP respectively. Conclusion The both of KM and ICR mouse were suitable for EFS cryoprotectant. EFS and DAP cryoprotectant could be used to froze embryos of C57BL/6J、OB/OB、LAP/rtTA0F59 mouse strain, then DAP was superior to EFS. But it is necessary that the further study on embryos cryopreservation in BALB/c mouse in future.

Abstract:SUMMARY: Objective: Histopathological examination of testes is important in assessing reproductive damage of drugs from toxicity studies. Superior fixative is an important factor for keeping stabilization of histological features. Method: Rat testis fixed in 10% buffered formalin (NBF), 10% NBF and Bouin’s fluid, Bouin’s fluid and Modified Davidson’s fluid (mDF), we compared the morphology of sections using hematoxylin and eosin (HE)-stained, Periodic acid-Schiff (PAS) staining and immunohistochmical (IHC) staining. Result: Fixation in mDF resulted in more sharply defined nuclear detail and better preservation of cellular cytoplasm and noticeably less shrinkage of the seminiferous tubules and superior overall morphologic detail compared to other fixatives. And the mDF also supported excellent staining of acrosomes with Periodic PAS staining reagent and superior detection of androgen receptor with IHC.

Abstract:Objective To investigate the effects of Shenwu capsule (SW), a new combination drug of traditional Chinese medicine, and its component tetrahydroxystilbene glucoside (TSG) on the expression of never growth factor (NGF) and its receptor in the hippocampus of aged rats. Methods The aged rats were intragastrically administrated by SW (0.8g?kg-1 and 1.6g?kg-1) and TSG (0.03g?kg-1 and 0.06g?kg-1) for 3 months (from 21 to 24 months of age), respectively. Real-time PCR was applied to measure NGF mRNA and TrkA mRNA in the hippocampus. Immunohistochemical staining and Western blot methods were used for detecting the expression of NGF and TrkA in the hippocampus. Results From the levels of gene and protein, the mRNA and protein expression of NGF and TrkA showed a decrease in the hippocampus of aged rats. Intragastrical administration of SW to the aged rats up-regulated the expression of NGF mRNA and TrkA mRNA in the hippocampus; SW and TSG enhanced the expression of NGF, and the high dose of SW and TSG elevated the expression of TrkA. Conclusion SW and TSG increased the expression of NGF and its receptor in the hippocampus of aged rats, suggesting that the drugs may act on the beginning of neuronal signal transduction pathway, and may be beneficial to neuronal survival and function.

Abstract:The pathological changes of dendritic spines and synapses plays an important role in cognitive impairment, and development regulation brain protein(Drebrin) can regulate the morphology and plasticity of dendritic spines and synapses. The decrease of Drebrin can affect the morphology of dendritic spines through the changes of spine cytoskeleton, and it can lead to changes in synaptic structure and function. But specific mechanisms of synaptic pathology changes in brain and the relationship between Drebrin and synapses in Alzheimer’s disease(AD) still remain unclear. Exploring the relationship between Drebrin and cognitive function and internal mechanisms has important significance to early prevention of cognitive impairment and find effective measures for diagnosis and treatment of AD.

Abstract:Objective： To explore the relevance of Wagner electrocardiogram QRS score to myocardial infarction area in diabetes mini-pigs. Methods： 12 Obama mini-pigs were randomly divided into diabetic group (n=6) and normal control group (n=6). The diabetes models were established by administering STZ of 150 mg/kg once intravenous. Fasting blood glucose were measured before and 1 day, 3 days and 7 days after drug administration. When fasting blood glucose consistently elevated (≥7.0 mmol/L), the model pigs were determined to be with diabetes. Second, selectively ligating the left anterior descending artery between diagonal branch 1 and 2 in diabetes and normal control mini-pigs, and perform QRS scoring at 10 min, 30 min, 1 h, 48 h after ligation; At the end point, the myocardial infarction size were measured by Evan 's blue and TTC staining; and the correlation of QRS score and myocardial infarction size were analyzed. Results: All animals happened typical pathological changes of myocardial ischemia; pathological Q wave formed, QRS scores in diabetes group was obviously higher than that in control group (6.9?.4 vs. 4.1?.8, P<0.05); Pathological staining results also showed that the infarction area was significantly larger than that in control group (29.2?.1% vs.15.3?.4%, P<0.05); the correlation coefficient was 0.92. Conclusion: Acute myocardial ischemia in diabetic mini-pigs were more likely to result in myocardial tissue necrosis, infarction area was larger; The myocardial infarction area judged by electrocardiogram QRS scoring had good correlation with pathological situations.

Abstract:Objective Using Morris water maze to evaluate the effect of curcumin on cognitive function of APPswe/PS1dE9 double transgenic mice. Methods 3-month old APPswe/PS1dE9 dtg mice were randomly divided into model group, Rosiglitazone group and curcumin large, medium and small dose group( n=6), with age-matched C57BL/6J mice of the same genetic background as normal control group. Medication was given once a day by intragastric administration for 6 months. Application of Morris water maze to test APPswe/PS1dE9 double transgenic mice spatial learning and memory ability. Results Curcumin to APPswe/PS1dE9 double transgenic mice spatial learning and memory disabilities have improvement effect, especially in curcumin-M dose group in the more obvious effects. Conclusion Curcumin can improve learning and memory ability of APPswe/PS1dE9 double transgenic mice.

Abstract:【ABSTRACT】 Objective To research the antitumor effection.of MIP-1α and B7-1 gene transfection. on mouse lymphoma. Methods Lentivirus-mediated mouse MIP-1α and B7-1 gene vectors . infected EL-4 cells. The MIP-1α和B7-1 mRNA were identified by RT-PCR and. MIP-1α和B7-1 protein were examined by Western blot. Then the infected EL-4 cells were injected into right armpits of mouse, and observed the development of tumor. The transfected EL-4 cells were infected to tumor-bearing mice, and observe the antitumor effection. Results RT-PCR assay showed that there were MIP-1αand B7-1 mRNA in the EL-4/MIP-1α+B7-1 cells, as well as MIP-1αand B7-1 protein examined by Western blot. MIP-1α and B7-1 group occupied much more time than the control group in the tumor generation, and which mean tumor volume was also smaller than the control group, as well as the rate of tumor formation, however there were no tumor generation in unite group. The average tumor volume，weight and organ involvement rate of MIP-1 α and B7-1 group was significantly smaller than that of the control group ( P < 0.05 ), while the average tumor volume and weight of united group was also significantly smaller than that of MIP-1 α and B7-1 group ( P < 0.05 ). In addition, the mean survival time of unite group was significantly longer than single group, and the control group ( P < 0.05 ). .Conclusion Lentivirus-mediated mouse MIP-1α and B7-1 gene transfection could effectively enhance the anti-lymphoma effection in tumor-bearing mice, and significantly prolonged the average survival time. Which also provided a new idea for the lymphoma gene therapy .

Abstract:【Abstract】 Objective To investigate the effect of microRNA-424 (miR-424) on neuronal apoptosis after cerebral ischemia in mice. Methods The lentiviral mediated overexpression of miR-424 (109U/ml, 8ul) was injected into the lateral ventricle in mice for 7d, then the middle artery occlusion (MCAO) model was established, and the mice were divided into 4 groups: sham group, sham miR-424 group, MCAO model group and MCAO miR-424 group (N=6). At 8h after MCAO, the ischemic brain tissue was removed and paraffin sections were used to perform TUNEL staining to observe neuronal apoptosis, and the protein levels of transcription factor PU.1, hypoxia inducible factor-1α (HIF-1α), apoptosis-related proteins p53 were detected by western blot. Results The TUNEL immunofluorescence indicates that miR-424 can reduce neuronal apoptosis at 8h after cerebral ischemia in mice, and western blot showed that, before ischemia or at 8h after ischemia, the effect of miR-424 on three proteins expression is the same for normal or MCAO model brain tissue, increasing the levels of PU.1, HIF-1α and apoptotic protein p53. Conclusion miR-424 can reduce neuronal apoptosis after cerebral ischemia partly through increasing expression of transcription factor PU.1 and HIF-1α, as well as apoptosis-related proteins p53 in the brain tissue of mice.

Abstract:【Abstract】: Objective: To establish a method for the detection of mRNA transcription of SLAM as CDV receptors in different tissues of canine. Method: By using GAPDH as the house keeper, The △△Ct relative quantification method was used to detect the transcription levels of SLAM mRNAs in different tissues of canine. Result: The inter-assay variability (CV %) were0.89 %- 2.35 %. The results indicated that the qPCR had advantages of high reproducibility. Results of the study showed that the SLAM mRNA were transcripted at high level in the spleen, hilar lymph node, lymphonodi mesosteniales, inguinal glands and low level in the urinary bladder. Conclusion: The quantitative RT-PCR method was established successfully for the detection of mRNA transcription of SLAM in different tissues of canine.

Abstract:Objective To observe myocardial infarction area, cardiac troponin -T (cTn-T) expression, expression of Bax, Bcl-2 mRNA of blueberry anthocyanins (Blueberry anthocyanin BBA) pretreatment on experimental acute myocardial infarction in rats and explore the intervention of the mechanism of myocardial infarction. Methods 40 Wistar rats were randomly divided into sham operation group, myocardial infarction model group, BBA low, middle, high dose group, drug intervention for 4 weeks, after 30 minutes of the last administration, ligate branch ligation of the left anterior descending coronary artery to establish myocardial infarction animal model. After 24 hours, TTC detection of myocardial infarction area; Western blotting detection of myocardial cells cTn-T protein expressing; expression of real time PCR for detection of Bcl-2 mRNA, Bax mRNA. Conclusion Protective effect of blueberry anthocyanins on myocardial cells after myocardial infarction is clear, its mechanism may be related to reduce the area of myocardial infarction; up-regulate the expression of cTn-T protein, up-regulate the Bcl-2 mRNA expression, down-regulate the Bax mRNA expression, inhibite myocardial cell apoptosis after myocardial infarction.

Abstract:Alzheimer’s disease (AD) is a progressive and fatal neurodegenerative disorder. The aggregation of hyperphosphorylated tau into neurofibrillary tangles (NFT) is a hallmark in the histopathology of AD. A number of mouse models have been created to study the major neuropathological mechanism and the research on drugs. We provide here some considerations for selecting a mouse model of AD, including tau transgenic mice, hyperphosphorylated tau animal model iduced by imbalance of the activities of kinases and phosphatases, hyperphosphorylated tau animal model induced by decreasing O-GlcNAcylation.

Abstract:【Abstract】Objective To research the characteristics of SHIVKU-1 infection in Chinese rhesus macaques and to provide an animal platform in evaluation of the effectiveness of HIV-1 drugs or vaccines with SHIVKU-1/rhesus animal model. Methods Two healthy Chinese original rhesus macaques, serum negative to SIV, SRV and STLV-1, were infected with SHIVKU-1 intravenously. CD4 and CD8 T cell subsets including memory CD4 T cells subsets in peripheral blood (PBL), inguinal lymph nodes (ILN), intestinal lamina propria (LP) were analyzed by ?ow cytometry. Viral loads was detected by real time RT-PCR, so was the specific IgG antibody titer against SHIVKU-1 in plasma using ELISA. ResuLts Both of the two rhesus macaques established systematic infection, keeping the steady viral loads in 3 months after infection. G1302V died 457 days post infection. Peripheral CD4 T lymphocytes decreased significantly post infection, while CD4 / CD8 went upside down. The proportion of CD4 Tcm cells experienced a decline in early stages of infection and rose sharply, especially in the peripheral blood and lymph nodes. CD4 Tem increased significantly in the intestinal lamina propria. Conclusion In this study, the establishment of SHIVKU-1 infection model of Chinese rhesus macaques provided us a choice to evaluate the efficiency of the candidate vaccines or the drugs aiming at the X4 tropic HIV, so did for the pathogenesis studies as well.

Abstract:Objective: To generate and identify the mammary gland Scrib gene specific knockout heterozygous mice by Cre-LoxP recombination system, for the research of the role of the Scrib gene on breast cancer at the whole animal level. Method: The heterozygous Scrib conditional knockout was generated and identified. The positive offsprings were mated with MMTV-Cre homozygous mouse, which express Cre recombinase in mammary epithelial cells. The gene type of the offsprings were identified. Results: The genetation of the Scrib heterozygous mice and MMTV-Cre mice were successful and Scrib /fl mouse were identified. 5 Scrib /fl;MMTV-Cre /- mouse were acquired through mating Scrib /fl mouse to MMTV-Cre mouse. Conclusion: Generation of mammary epithelial cells Scrib gene knockout heterozygous mice by Cre-LoxP recombination system was successful, which can supply a novel animals model for further research of the polarity protein Scrib down-regulation on breast cancer．

Abstract:Objectives: To construct recombinant adeno-associated Virus and lentivirus carrying siRNA of TIMP-1 and to investigate the antifibrotic effects of these two recombinant viruses on CCl4 induced liver fibrosis in rats. Methods: One pair of siRNA which could effectively inhibit expression of the TIMP-1 gene in HSC-T6 was screened and cloned into AAV vector and lentiviral vector to construct the recombinant AAV/siRNA-TIMP-1 and Lenti/siRNA-TIMP-1. AAV/EGFP and Lenti/EGFP as negative control were also obtained. Wistar rats were randomly divided into six groups: Control group, CCl4 group, AAV/EGFP, Lenti/EGFP, AAV/siRNA-TIMP-1 and Lenti/siRNA-TIMP-1 group. After four weeks administration of CCl4, liver samples were collected for the histochemical stain and the detection of TIMP-1 expression. Results: Livers from control rats showed normal H&E and Masson staining around vessels. In contrast, livers from the model, AAV/EGFP and Lenti/EGFP groups showed severe fibrosis, including septal fibrosis, extensive bridging, fatty degeneration. The mRNA and protein expression of TIMP-1 were also elevated in the livers from these groups. Compared with the fibrosis model group, the AAV/siRNA-TIMP-1 and Lenti/siRNA-TIMP-1 group showed good preservation of liver acini architecture and only mild bridging fibrosis, accompanied by the decreased mRNA and protein expression of TIMP-1. Semi-quantitative analysis of the fibrosis stage indicated that most rats in the model, AAV/EGFP and Lenti/EGFP groups were at S3 and S4 (80%), while 20% of rats were at S5. In contrast, most rats (90%) in the AAV/siRNA-TIMP-1 and Lenti/siRNA-TIMP-1 group were at stages S2 and S3, with only one at S4. There was no significant difference between these recombinant virus therapy groups. Conclusions: AAV/siRNA-TIMP-1 and Lenti/siRNA-TIMP-1 could suppress the expression of TIMP-1 in rat fibrotic liver, which may be effective antifibrotic gene therapy agents.

Abstract:【Abstract】 Objective To study the potential role of FKBP52 in the development of prostate in FKBP52 knockout (KO) mouse model. Methods The prostates of WT and FKBP52 KO were collected at different stage, E17.5, P0 and 3-week. HE staining was used to observe the morphogenesis of prostate at different periods of prostatic development. Results It was found that FKBP52 deficient mice still can initiate the prostatic lobes. Branching morphogenesis is almost entirely complete by 3 weeks after birth in the WT mouse. In FKBP52 KO mouse, however, at the same time the prostate is totally underdevelopmental. Conclusion FKBP52 plays an important role in the postnatal development and its function can influence the maturity of prostate.

Abstract:Shower of dressed in protective clothing is an important part to ensure laboratory safety during work in the high level of bio-safety laboratory. The article introduce the chemical shower facilities and the equipment from several aspects such as structure、technical skill、characteristics and so on. The article also discuss about the necessity of domestic chemical shower technology research and several issues should be focused on during the development process.