Abstract:Objective To observe the sterilization effects of hydrogen peroxide vapor on hard surface and air in laboratory animals barrier environment. Methods BIOQUELL Z sterilizer continuously sprayed 30% hydrogen peroxide vapor by 6 mL/m3、9 mL/m3 and 12 mL/m3 for 60 min at different positions in laboratory animal barrier environment. The numbers of falling bacteria and bacteria on the surface in barrier environment were detected. Rusults The falling bacteria in second changing rooms, clean corridor, storage room, dirt corridor and buffer room were killed completely by amount of 6 mL/m3 H2O2, and the falling bacteria in animal feeding room were killed completely by amount of 9 mL/m3 H2O2, but the bacteria on the surface in barrier environment were not completely killed. The bacteria on the surface in barrier environment were completely killed by amount of 12 mL/m3 H2O2. Conclusion Hydrogen peroxide vapor has very good sterilization effects on laboratory animal barrier environment and housing facilities.

Abstract:Intrauterine growth restriction(IUGR) is the one important complications in the perinatal period, and associated with poor perinatal health outcomes. In animal fetuses, IUGR can be caused by factors that are of maternal, placental or fetal origin.The most common cause of IUGR is placental insufficiency. However, there is a range of issues that require further investigation or investigation in more than one models. The information collected different animal models of IUGR may contribute to investigating its pathogenesis and seeking effective ways. In this article, the authors review the experimental animal models of IUGR, including natural selection, the surgical ablation of the majority of endometrial caruncles, passive smoking, matemal undernutrition, ligation of an umbilical artery.

Abstract:Hand Foot and Mouth Disease (HFMD) is caused by enterovirus family, in which enterovirus 71 (EV71) is the most common pathogen.Serious cases lead to neurological sympotoms and even death, especially in infants. A good EV71 infection animal model will not only help to study the mechanism of pathogenesis, and will become an effective tool for developing vaccines and treatments. Here we review advances in establishment of EV71 infection animal models in different species, and further focuse on states and prospects of EV71 infection models established by genetically modification.

Abstract:Objective In view of better performance of rat than mice in nervous diseases studies, to generate a transgenic Parkinson's disease rat line expressing alpha-synuclein gene with A53T mutation and provide a better PD animal model. Methods The transgenic plasmid was constructed by inserting the alpha-synuclein gene with A53T mutation into the downstream of mouse PDGF promoter. The transgenic rats were produced by microinjection and the genotype was detected by PCR. The protein expression levels in brain tissues were determined by western blotting. The expression of human alpha -synuclein in the brain tissues from transgenic rats was analyzed by immunohistochemistry staining(IHC) and immunofluorescence(IF)．The changes of the number of tyrosine hydroxylase(TH)-positive neurons in the substantia nigra were observed by IHC and IF. The motor performances of transgenic rats were measured by rotating rod experiment and gait analysis system. Results One transgenic rat line with high human alpha-synuclein expression levels was established. Histopathological examination revealed the abnormal aggregation in midbrain dopamine (DA) neurons and significantly reduced number of TH-positive neurons （69%，P<0.05) in substantia nigra from transgenic rats. Rotating rod and rat gait analysis indicated declined motor performances（40% - 60%，P<0.05)） and gait disorder of transgenic rats. Conclusions The human a-synuclein A53T transgenic rat models of Parkinson’s disease were established．

Abstract:Objective Establishment a Sleeping Beauty (SB)transposase-expressing transgenic mice for study of the genetic modification mediated by transposon system in mice. Methods Using the UBC promoter to drive the expression of SB transposase. The transgenic mice were created by microinjection. The gene type of transgenic line was identified by PCR. The expressing level in testis was determined by Western Blot and immunohistochemistry (IHC) staining. Results Four lines of transposase transgenic mice were obtained by microinjection and three can be germline. One mouse line with higher expression level of transposase in the testis was obtained. Conclusion One transgenic mice model with Sleeping Beauty transposase –expressing was successfully established. This model will greatly contribute to the research of genetic modification mediated by transposon in mouse.

Abstract:【Abstract】Objective To evaluate the effect of RNA expression level of tyrosinase family on rabbit (Oryctolagus cuniculus) coat and iris colour. Methods RNA were extracted and RNA expression level of Tyr, Tyrp1, and Dct were examined in skin and iris of 48 rabbits from 4 breeds including WHBE, JW rabbit, Rex rabbit and Chinchilla rabbit. Results Tyr expression in the iris of WHBE and Chinchilla rabbits significantly increased, as did Tyrp1 expression in the skin of Chinchilla rabbits. No significant different were found between the 3 genes in the other tissues. Conclusion Our results indicated the gene expression levels of Tyr1 in dark skin tissue were significantly higher than that in white one. Gene expression levels of Tyr in dark iris tissue were significantly higher than that in colourless iris.

Abstract:【Abstract】 Objective To generate and identify the AD model heterozygous mice by hybridization with the mice over-expression of Slit2 protein, for the research of the role of the over-expression of Slit2 protein on AD at the whole animal level. Method The heterozygous Tg2576 transgenic mice was generated and identified. The positive offspring were mated with Slit2-Tg homozygous mouse, which Slit2 protein was over-expression . The gene type of the offspring express were identified. Results The generation of the Tg2576 heterozygous mice and Slit2-Tg mice were successful and Tg2576 /- mouse were identified. 5 Tg2576 /-;Slit2-Tg mouse were acquired through mating Tg2576 /- mouse to Slit2-Tg mouse. Conclusion Generation of Slit2 protein over-expression heterozygous mice by hybridization with AD model heterozygous mice was successful, which can supply a novel animals model for further research of the Slit2 protein over-expression on AD．

Abstract:Aim To assess the sustainability of visceral hypersensitivity model induced by rectal distention with limbs constraint. Methods Thirty-two 8-week Sprague-Dawley(SD) rats were randomly divided into control group (n = 16), model group (n = 16), rectal distention with limbs constraint lasting for 2 weeks was operated on model group, visceral sensitivity was evaluated by abdominal withdrawal reflex(AWR). Based on the AWR score, control group were randomly divided into group A (n = 8) and group B (n = 8), model group were randomly divided into group C (n = 8) and group D (n = 8). Colon specimens of group A and group C were obtained Under anesthesia. Group B and group D received normal breeding until 18 weeks old, and then group D received the same stimulation. AWR score and colon specimens were obtained in the same way. ultrastructure of mast cell was observed by electron microscopy, toluidine blue staining was performed to count mast cell(MC), protease activated receptor 2 (PAR2) was analyzed by Western Blot. Results AWR score of group C was higher than that of group A, group D lower than group C, the differences had significance (P < 0.01). Intestinal MC count of group C was higher than that in group A, group D lower than group C, the differences had significance (P < 0.01). Mast cell degranulation was visible in group C, neither other groups. PAR2 expression level of group C was higher than that of group A, group D lower than group C, the differences had significance (P < 0.01). Conclusion Visceral sensitivity of visceral hypersensitivity model induced by rectal distention with limbs constraint decreased along with the increase of weeks.

Abstract:Objective To construct an adenovirus vector containing eNOS gene，and analyze the expression and location in endothelial progenitor cells from bone marrow. Methods Establish the vector for recombinant adenovirus: the recombinant plasmid PSUCMV-heNOS was established by inserted the heNOS gene in the vector PSUCMV. After identified for correction in sequence, it was transferred to cell 293 with the plasmid including the right arm of 5-type adenovirus by Lipofectamine 2000. EPCs from bone marrow transfected by recombinant adenovirus with eNOS gene: 18 male Wistar rats, aged 4-6w, were elected for EPCs culture. The cells in the third passages were randomly allocated to 3 groups: PBS control group (Cgroup), Lac gene transfection group(Lgroup) and the eNOS gene transfection group(N group). Every group was separated into 3 subgroups following the 1d, 3d and 7d after treatment. The changes in the morphology, the product of NO in the culture medium and the express of eNOS were monitored. Results The DNA of AdCMVheNOS was successfully transferred into the 293 cells by liposome and packaged inside. After confirmed by PCR analysis, the titer was 1.58?010PFU/ml after amplified. The culture of EPCs from bone marrow in rat was observed, and the expression of eNOS product was increased after transfected for 1 day. At the same time point, the content of eNOS in N group was significantly higher than the L group and C group. The N7 and N3 subgroup showed a significant difference compared with the N1 subgroup. The production of NO increased following the time and the difference was statistic between the 3 day and the 7 day subgroups. Conclusion Adenovirus can mediate the high quality expression of eNOS gene in the EPCs and induce the increasement of NO in culture medium and eNOS expression in cells.

Abstract:Objective To investigate the mechanism of expression of TNFα of renal tubular epithelial cells with hypoxia reoxygenation (H/R) and to evaluate the effects of extracts of Picorhiza scrophlariiflora Pennell on it. Methods The cutueled renal epithelial cells were divided into groups: normal control grounp,H/R group, 10μg/ml extracts of Picorhiza scrophlariiflora Pennell H/R group, 100μg/ml extracts of Picorhiza scrophlariiflora Pennell H/R group, 1000μg/ml extracts of Picorhiza scrophlariiflora Pennell H/R group. The flow cytometry was used to detect the oxidative stress as well as expressions of TNFα protein.Real time quantitative reverse transcription-polymerase chain reaction(RT-PCR) was used to detect expression of mRNA for TNFα. Result The oxidative stress in renal tubular epithelial cells was induced after H/R. TNFα protein were increasd after H/R. TNFα mRNA were increasd after H/R.Oxidative stress in renal tubular epithelial cells was inhibited by extracts of Picorhiza scrophlariiflora Pennell with a dose-dependent manner. TNFα protein were inhibited by extracts of Picorhiza scrophlariiflora Pennell with a dose-dependent manner. TNFαmRNA were inhibited by extracts of Picorhiza scrophlariiflora Pennell with a dose-dependent manner. Conclusions Extracts of Picorhiza scrophlariiflora Pennell protected renal epithelial cells against TNFαby inhibiting oxidation after H/R injury.

Abstract:Objective To explore the Effect of remote ischaemic preconditioning on renal function in rats with renal ischemia reperfusion. Method Methods: SPF male SD rats (n=30) were randomly divided into three equal groups: sham group (group S), ischemic reperfusion group (group IR), remote ischaemic preconditioning group (group RIPC). For group IR, cutting off the right kidney, and lefting the right one ischemia for 45min, then transfusing blood for 60min. For group RIPC, occlusion femoral artery for 5min, reperfusion 5min, repeated three times, making IR models after 1h. For group S, we lift right kidneys after anesthesia and laparotomy. Blood sample was collected in 6 h to Scr，BUN，IL-1β，TNF-α and Kim-1. Histopathological changes of kidney were observed by using of pathological section and HE staining. Results: Histopathological observations of kidney shows no obvious change in kidneys of group S. Serious kidney injury was found in group IR, which showed up as obvious inflammation and glomerulonephritis, While the kidney injury was significantly alleviated in group RIPC, and no serious inflammatory response was detected. Compared to group S, levels of Scr，BUN，IL-1β，TNF-α and Kim-1 in group IR and RIPC had increased significantly (P <0.05); which had significant decreased in group group IR and RIPC (P<0.05). Conclusion RIPC can reduce inflammatory reaction, thus alleviating renal ischemia reperfusion injury and protecting kidney to some degree.

Abstract:[Abstract] Objective To establish a real-time RT-PCR method for detection of simian parainfluenza Virus 5 type (SV5) in biological material from monkeys and explore its application．Methods According SV5 virus NP gene fragments, a pair of specific primers and TaqMan probe were designed and synthesized; SV5 RNA standards were prepared using some molecular biological methods. Then the linearity, sensitivity, specificity and stability of the method were measured. Last, the method was used for testing lung samples of 24 monkeys. Results we established successfully SV5 real-time PCR method. The linear range of 6.8 ?109 copies/μL ~ 6.8 ?105 copies/μL, a sensitivity of 6.8 copies/μL, coefficient of variation (CV) was 0.63% ~ 8.71%, inter-assay CV 1.52% - 12.38%, and the method is high specific. There wasn’t detected positive in lung samples of 24 monkeys. Conclusions Establishment of the method provides the SV5 quantitative detection assay for the experimental animals and animal-derived biological products.

Abstract:Objective To make sure the mutation sites of simian immunodeficiency virus (SIVmac239) resistance to 9-(2-methoxy propyl phosphate) adenine (PMPA), to guide the using of SIV/rhesus monkey model in the evaluation of HIV-1 drug. Method Rhesus macaque (RM) were infected with a RM-adapted SIVmac239 strain, were treat with PMPA. And plasma viral load and peripheral CD4 T cells was measured at different time points. Then the drug sensitivity of the virus strain was detected using CEMx174 cells. At last, we screened the mutation sites of rt gene from the virus strain by single genome amplification. Results PMPA treatment can’t effectively reduce the plasma viral load and peripheral CD4 T cells count had no significant chance. Drug sensitivity experiments confirmed that the virus strain is of insensitive to PMPA. Sequence alignment found that S205L and M502I of viral rt gene may affect PMPA sensitivity. Conclusions This study provided some information for the rational use of SIV/rhesus monkey model and clinical treatment of HIV-1.

Abstract:Objective To Comparative study of three anesthetic methods in hemorrhagic shock experiments of Bama Minipigs. Methods Bama Minipigs 24, the average into three groups are Intramuscular ketamine group (A group), intravenous propofol group (B group), propofol intravenous anesthesia with sevoflurane group (C group). Respectively, before anesthesia (T0), after induction (T1), after catheter (T2), HS instantly (T3), HS after 30min (T4), recovery (T5), resuscitation after 30min (T6) and pull the end of the experiment tube after 10 min (T7), pigs in each group were observed and recorded heart rate (HR), mean arterial pressure (MPAP), nasal temperature (NT), pH value and oxygen saturation (SPO2) and other indicators, while recording the induction of anesthesia time, restoration of spontaneous breathing time, intraoperative anesthetic additional number of cases, number of cases of postoperative adverse reactions. Results Three groups of Bama pig experiments predecessor length, weight, volume and put the blood hemoglobin general condition showed no significant difference (P> 0.05); A group of anesthesia to be ineffective, surgery and have to be repeated dosing restless phenomenon surgery after vomiting and other adverse reactions; B group anesthesia is relatively stable, but still restless phenomenon during surgery and need additional anesthetic; C group anesthesia best short induction period, intraoperative safety, restoration of spontaneous breathing short time, intraoperative no adverse effects. Conclusion Propofol intravenous anesthesia combined with sevoflurane anesthesia induction time is short, the anesthetic effect is good, smooth and safe surgery, postoperative recovery fast, suitable for large animal anesthesia.

Abstract:Objective Using behavioral testing to study the influence of simulated astronautic environment on cognitive function in rats.Methods Tail suspension, separation, isolation and circadian rhythms change are used to simulate the stronautic environment. Reward conditioning test and Morris water maze (MWM) test are used to measure cognitive function after 14days and 28days of simulated compound astronautic environment. Results Upon reward conditioning test, compared with control (Con) group , the compound environment (CE) group showed the lower nose pokes(P＜0.05) in 14 days. In 28d, CE group was significantly less than Con group and Tail restraint(TR) group in nose pokes(P＜0.01). Upon the Morris water maze(MWM) test , compared with Con group and TR group, CE group exhibited longer escaping latency and total distance in 14d (P＜0.01，P<0.05). At 28d, the difference became larger (P＜0.01). Con group and TR group were no difference in reward conditioning test and MWM test. Conclusions Simulated compound astronautic environment can induce serious cognitive function deficit, and the cognitive function deficit was more stable in 28d.

Abstract:Objective Atfu-B*sv1 with α2 domain deletion is a splice variant of major histocompatibility complex class I B in Ateles fusciceps. In this experiment the basic information about expression and cellular location of Atfu-B*sv1 was got, and aimed at compared with the full-length Atfu-B genes, observed the different from them. Methods Eukaryotic expression vectors of Atfu-B*sv1、 Atfu B*02:03 and Atfu-B*03:01 were constructed and transfected to 293T cells. The expressions of the three genes in cells were detected by Western blotting methods, and the cellular locations of the three genes were analyzed by immunofluorescence and laser confocal microscopy. Results Atfu-B*sv1、Atfu-B*02:03 and Atfu-B*03:01 were all expressed in the 293T cell and were glycosylation . All the three genes were expressed on the cell membrane. Conclusion The expression and cellular localization of Atfu-B*sv1 was not significantly different from the full length Atfu-B genes. Further researches about functions of Atfu-B *sv1 need to be done.

Abstract:Objective To study the effect of SIVmac251-infection on apoptosis induction of macrophage from rhesus macaque by galectin-3. Methods Monocytes were isolated from peripheral blood of healthy or SIVmac251-infected rhesus macaques. Cells were induced to differentiate to macrophages by GM-CSF and IL-6. Macrophages were cultured for 5.5 hours at the presence of galectin-3. Cells were stained by PE-Annexin V/ PerCP-7ADD and apoptosis was examined upon flow cytometry. Results The percentages of Annexin V cells without galectin-3 induction were 5.08% and 4.87% for macrophages from healthy or SIVmac251-infected rhesus macaques respectively. The percentages of them with galectin-3 induction increased to 17.62% and 45.56% respectively. The increase for SIVmac251-infected rhesus macaque was significantly greater than that for healthy monkey (p﹤0.01). Conclusion SIVmac251-infection results in sensitivity enhancement to apoptosis induction of macrophage from rhesus macaque by galectin-3.