Abstract:Objective To prepare the rabbit polyclonal antibodies against sixteen stains of birds，and to label the secondary antibodies with horseradish peroxidase for developing the serological test system of birds． Methods IgYs of birds were purified by water diluted，combining precipitation with ammonium sulfate，or affinity chromatography and precipitation with ammonium sulfate，or precipitation with ammonium sulfate and cutting the SDS-PAGE gel slices that contain the IgY bands． Rabbit anti-sera were prepared by repeatedly immunized rabbits with purified IgYs． The antibodies against bird IgY in rabbit sera were detected by agarose double immunodiffusion and purified using Protein A affinity chromatography． The products were labeled using HRP by periodate oxidation method． And the activities of the labeled rabbit polyclonal antibodies against sixteen strains of birds were detected by ELISA． Western blots were performed to investigate the specificity of IgG-HRP against the IgY of birds． Results The IgYs from the strains of birds were purified，including greylag goose，great cormorant，ostrich，little grebe，pigeon，rich bird，goose，peacock，quail，ladyhood-chicken，heron， night-heron，red-crested pochard，tern，black-winged stilt，parrots，common shelduck． The titers of sixteen rabbit antisera were up to 1 ∶ 32 examined by agarose double immunodiffusion． Sixteen kinds of purified rabbit anti-bird secondary antibodies were labeled with HRP and their activities were up to 1∶ 800 － 80000 examined by ELISA． The specific activities of the sixteen rabbit anti-bird IgGs-HRP were indicated with western blot． Conclusions Rabbit polyclonal antibodiesagainst sixteen birds labeled with HRP were prepared successfully; It was an useful tool for developing the serological test system for birds．

Abstract:Objective To investigate the effect and probable mechanism of icaritin ( ICT) on EL-4 cells in vitro．Methods MTT、transmission electron microscope ( TEM)、flow cytometry analysis were used to examine the effect of ICT on the proliferation of EL-4 cells． Reverse transcript-polymerase chain reaction ( RT-PCR) assay and colorimetric method was used to reveal the probable mechanism． Results ICT could inhibit the proliferation of EL-4 cells significantly in a dose-and time-dependent manner． Apoptotic cells were observed by TEM and flow cytometry analysis． RT-PCR analysis showed that the expression of bcl-2、P21 were down-regulated． Besides ，Caspase-3 and Caspase-9 activity in EL-4 cells elevated remarkably． Conclusions ICT could inhibit the proliferation of EL-4 cells and induce EL-4 cells apoptosis in vitro． The probable mechanism may be down-regulate the expression of bcl-2、P21and elevate Caspase-3 and Caspase-9 activity in EL-4 cells．

Abstract:Objective To consummate genetic biochemical marker of inbred mice，we study Catalase-2 of biochemical markers of inbred mice． Methods Filial generation 1 CCBF1 were abtained by intercrossing of CBA /Ca and BALB / c; and filial generation 1 B6CBF1 were also abtained by intercrossing of CBA /Ca and C57BL /6． Filial generation 2 were obtained by intercrossing between filial generation 1 and by backcrossing between filial generation 1 and their parents．Then filial generation 2 were monitored by biochemical markers． Result Ce-2b is detected among filial generation 2 mice，while not among filial generation 1 mice． Conclusion Ce-2a is fully dominant．

Abstract:Objective To analyse breeding and genetic stability in closed Colony of uncleaning Mongolian Gerbil (Meriones unguiculatus ) ． Methods Fourty pairs Mongolian Gerbil were selected randomly，recorded amount about reproduce and growth． Thirty three were selected for analysis genetic stability． Mitochondria D-Loop were amplified respectively by using PCR of a pair primers． The results from DNA sequencing were compared with the standard sequence of Mongolian Gerbil． Results The average 7． 01 Mongolian Gerbil were reproduced in one time． Male is higher than famale in body weight． The sequence of D-Loop gene was same in thirty three Mongolian Gerbil． Conclusion Polymorphism of mitochondrial DNA was not found． The Mongolian Gerbil colony showed good genetic stability．

Abstract:Objective This study is aimed to investigate the effect of soy isoflavones on initial oestrus and some physiological characteristics of female mice． Methods Total two hundred and ten 3-week old female ICR mice were randomly divided into 3 groups，and fed with the control (0 mg / kg) ，low soy isoflavone level (50 mg / kg) and high soy isoflavone level ( 400 mg / kg ) diet separately． The state of vagina opening and vagina plug after treatment with soy isofalvones，the concentration of estrogen in their serum on Day45 and Day65 as well as their body，uterus and ovarian weight， were analyzed and recorded separately． The ER ( estrogen receptor ) on uterus was detected by immunohistochemical staining． Results There were significant differences of the durations for murine vagina opening between those treatment groups and control one． The body weight of two treatment groups was significantly lower than that of the control one． The positive rate of ER on the stroma in control group was significantly lower than that of the other treatment ones，but that on the glandular epithelium in high level group was much higher than the other groups’，and there were no significant differences on the luminal epithelium among all the groups． Conclusions Collectively，it can be concluded that the 400mg / kg soy isoflavones in the diet can simulate murine vagina opening，increase their body weight and degrade the estradiol level in their serum on Day65． affect the expression of estrogen receptor on their utrine stroma， luminal epithetlium after 8 weeks． however，50mg / kg soy isoflavones in the diet only affect the body weight of 45d，and the expression of estrogen receptor on their utrine stroma after 8 weeks

Abstract:Objective To establish the background database of non-proliferative spontaneous lesions of F344 rats for drug safety evaluation． Methods SPF F344 rats 336，male and female in half，8 months old，kept under the center barrier，were randomly divided into four groups，animals were killed32，64，64，176 at respectively 6，12，18 and 24 months time points． Tissues were examined under microscopic，and statistics 4 time points observed spontaneous animal disease type and disease incidence． Results Non-proliferative lesion consists of murine progressive cardiomyopathy，BALT，tubular lesions，and genital tissue degeneration such as reduction of ovarian cells or inhibition of spermatogenesis．Conclusions Under the condition of center barrier，the incidence of non-proliferative lesions of F344 rat increased． The present study enriched the existing F344 rat pathology background information，and laid the foundation for carrying out drug safety evaluation and research．

Abstract:Objective Swrodtail，Xiphophorus hellerii was an aquatic experimental animal authorized by the original thoroughbred committee and published by the ministry of agriculture． It shows a good prospect in genetic research experiment，water environment pollution monitoring and bacterial disease research． In order to monitoring the germplasm resources of swordtail，distinguish the inbred lines and wild swordtail and identify the purity of inbred lines，microsatellite DNA fingerprint was used in this study． Methods 50 pairs of microsatellite primers were synthesized according to related reports． Among which，train-special primers used to identify the inbred lines of swordtail and core primers used to draw the DNA fingerprinting schema graph were screened． Then we constructed the DNA fingerprint using scatter diagram method with EXCEL 2003 software． Digital fingerprint barcode data had input software of PRIFRI V1． 0，which had formed a standardization fingerprint identification database of swordtail． Results We together received five pairs of strain-special microsatellite primers and 46 core primers． The DNA fingerprinting of RR-B，RW-H and wild swordtail were constructed．And a fingerprint identification database of swordtail was formed． Conclusion Using these microsatellite markers and DNA fingerprinting，we can identify varieties，purity and genetic monitoring of RR-B，RW-H and wild swordtail．

Abstract:Objective To explore the roles of antigen presenting cells (APC) in the lung of chronic inflammation by investigating the effects of smoking on the expression of B7-1 /B7-2 and its associated ligand in the lung tissue of rats．Methods Wistar rats ( n = 30 ) were randomly divided into non smoking group and smoking 6 weeks group，12weeks group，the number of each group was ten． Immunohistochemistry was used to semi-quantitatively analysis the changes of B7-1 /B7-2 and its associated ligands expression on the cytomembrane of chronic inflammation cells which were on the pulmonary interstitium around airways of rats． Results The expression of B7-1，B7-2，CD28 and CTLA-4 in the lung tissue of the smoking 6 weeks and 12 weeks groups' rats were higher than those of nonsmoking group ( P ＜ 0. 01) ，with the smoking 12 weeks group higher than the smoking 6 weeks group ( P ＜ 0. 01) ． Their expressions were increased in time-dependent manner． Conclusions Smoking can induce increasing expression of B7-1 / B7-2 and its related ligands in the lung tissue of rats; the APC may play an important role in the process of chronic pulmonary inflammation．

Abstract:Objective Effects of Cu2 + and SDS on the survival rate of Bareneck goby larva were studied．Methods Eight Cu2 + and seven SDS were designed． Survival rate data was collected during the 96 h's experiment．Results showed that the effects of Cu2 + and SDS on the survival rate of Bareneck goby larva were significant ( P ＜ 0. 05) ．With the increase of the daily age，the survival rate of the larva was increased． Conclusion The results indicated thatBareneck goby was more fitting than the parallel aquatic used to do some toxicity examination．

Abstract:Objective To investigate the effect of IL-19 on airway smooth muscle cells ( ASMCs) proliferation in asthmatic rats． Methods The rat model of asthma was made through ovalbumin ( OVA) sensitization and excitation． Primary cultures of ASMCs were established for experiments． ASMCs were treated with 1 μg /L、10 μg /L 、100 μg /L IL- 19． Adopt flow cytometry and MTT method to detect the proliferation of ASMCs，and observe the influence of different concentrations of IL-19 on the proliferation of ASMCs． Results Compared with normal rats，the proliferation of ASMCs in chronic asthmatic rats and the percentage of cells at S phase was obvious increased． The percentage of cells at S phase of ASMCs in chronic asthmatic rats intervened by 10 μg /L、100 μg /L IL-19 was decreased and the proliferation of ASMCs was reduced compared with the asthmatic group，and there is difference between 10 μg /L IL-19 group and 100 μg /L IL-19 group． But these two indexes of ASMCs in chronic asthmatic rats have no obvious change after intervening by 1 μg /L IL-19． Conclusion L-19 may inhibit the proliferation of ASMCs in chronic asthmatic rats．

Abstract:Objective To study immunogenicity of fusion gene Ag85B-Esat6-HspX of Mycobacterium tuberculosis． Methods Forty BALB / c mice were randomly divided into four groups (10 per group) : the plasmid pcDNAHspX group，the pcDNA-Ag85B-Esat6-HspX plasmid group，the BCG group and the saline group． The immunization was repeated three times two weeks apart． A single dose of BCG (1 × 106 CFU /mouse) was injected subcutaneously． Mice in other three groups were administered intramuscularly in the hind limb． Total IgG levels were detected at 2，4 and 6 weeks after immunization． Meanwhile，cell-mediated immunity responses were examined two weeks after the final immunization as well． Result Mice immunized with recombinant plasmid expressing the Ag85B-Esat6-HspX fusion protein generated specific cellular and humoral immunologic response to the stimulation of Ag85B，Esat6 and HspX． Conclusion The fusion gene Ag85B-Esat6-HspX of Mycobacterium tuberculosis could be DNA vaccine to carry out the further research of protective efficacy．

Abstract:Objective To study the role of WIP1 gene in mouse B cell and T cell development． Methods The proportion of the B cell in bone marrow and T cell in the thymus were determined by the flow cytometry． Results Although the proportion of different subsets in B cell development was normal，the total number of B cell in bone number was decreased． WIP1 gene knockout mice had defect in thymus development，the proportion of CD8 /CD4 double negative cells was increased，but the CD8 /CD4 double positive cells was decreased． Conclusion WIP1 gene plays an important role in the development of B cell and T cell．

Abstract:Objective To study the preventive immunity effects of myelin-associated neurite outgrowth inhibitors (NGI) recombinant DNA vaccine on APP /PS1 co-transgenic Alzheimer's model mice． Methods The recombinant DNA vaccine was intramuscularly injected in APP /PS1 co-transgenic mice． Wild type ( WT) group，blank group and blank vector group as control． The behavioral tests were examined by the Morris Water Maze． Result The behavioral impairments of vaccine group were significantly approved in Morris Water Maze ( MWM ) ( P ＜ 0. 05 ) ． Conclusion Intramuscularly injected with NGI DNA vaccine attenuated behavioral impairment in a co-transgenic mouse model of Alzheimer's disease．

Abstract:Objective To generate the interleukin 34 ( IL34 ) transgenic mice and investigate its functions on mouse＇s immune system． Method The transgenic plasmid was constructed by inserting the human IL34 gene in the downstream of CMV promoter． The transgenic mice were produced by microinjection method and the genotype was detected by PCR． The expression level of the protein was determined with Western blotting． The pathologic changes of important organs were observed by histochemical stain． Result Two lines of transgenic C57BL /6J mouse of IL34 gene were established． The expression levels of the IL34 gene was higher than the endogenous IL34 in the spleen tissue． Histology analysis demonstrated the important organs of the IL34 transgenic mice such as brain，heart，liver，kidney，intestine et al were nomal except spleen． Compare with the wild type mice，the germinal center of spleen was more activity，the area of white pulp were larger． Conclusion The IL34 transgenic mice were established successfully． Over-expression of IL34 in the immune systerm needs further investigating

Abstract:Objective To compare three methods for setting up artificial airway ( AA) of rabbit． Methods 30 New Zeland White rabbits were chosen and randomly divided into three groups: Group BOI ( group blind orotracheal intubation) ;Group COI ( group classic orotracheal intubation) ; Group TO ( group tracheotomy) ． The time for setting up artificial airway，the achievement ratio for the first time and the incidence of complication were observed． Results The shortest time for setting up artificial airway is Group BOI，showing a significant difference compared with other two groups (P ＜ 0. 05) ． The highest achievement ratio is Group TO，then Group BOI，and the lowest Group COI(P ＜ 0. 05) ． The highest incidence ratio of complication is Group TO，then Group COI，the lowest is Group BOI． Conclusions Blind intubation can be the first choice for setting up artificial airway for rabbit compared with classic oratracheal intubation and tracheotomy．

Abstract:Simian adenovirus( SAdV) ，as common viral agent persistent in respiratory and gastrointestinal tract of non-human primates，can cause pneumonia，gastroenteritis and conjunctivitis and spread mainly by Fecal-oral way． SadV can infect macaques，chimpanzees，baboons，etal belonging to genus Mastadenovirus ． Some resarchers agured that SadV had strict species specificity． Other reports suggested that cross species transmission may be a common occurance and is zoonotic transmission． No report says SadV can infect human，but found a novel adenovirus serotype that may be recombination with a human adenovirus． SadV and HadV are close，casuing similar clinical symptoms，which make it a good model for adenovirus research． To date，the goal of developing a non-human primate adenovirus model has been made to study the mechanisms of infection and promote progress on gene therapies and genetic vaccines with adenovirus vector． However，little is known about the prevalence of adenovirus infection in non-human primates． This article reviewed discovery and classification，biological characterization，and detection of SAdV so as to provide some useful insights for researchers．

Abstract:Through post-transcriptional regulation，microRNA can affect the expression of the proteins in the cell．MicroRNA play an important role in neuronal development，differentiation and normal function． In Alzheimer's disease the expression of microRNA is changed，the deregulated microRNA can affect the development of the disease in many aspects Study on the role of microRNA in Alzheimer's disease not only benefit exploring the pathogenesis of the disease but also will lead to better understanding of the regulation effect of microRNA．

Abstract:Heparin-binding epidermal growth factor-like growth factor ( HB-EGF) is a member of the epidermal growth factor (EGF) family． HB-EGF is a potent mitogen for a number of cell types and has been implicated in a variety of physiological and pathological processes，including myocardial remodeling，fibroblast hyperplasia and collagen expression． In this review，the HB-EGF in myocardial remodeling was summarized．

Abstract:This article summarizes some experience in teaching eight-year program medical students laboratory animal science and puts forward suggestions of related teaching method，teaching content，teaching focus，etc． The author hopes to construct a bridge for the future scientific research of eight-year program medical students and cultivate the outstanding scientific innovative ability of them．