Abstract:Objective To clone and analyze the whole sequence of mitochondrial cytochrome b (cyt b) gene of swordtail fish ( Xiphophorus helleri ).Methods\ Total DNA from the liver tissue of swordtail fish was extracted and a pair of specific primers were designed and processed for PCR amplification.Analyzed by agarose gel electrophoresis,the amplification products were purified and cloned into T vectors of pGEM\|T easy vector system.With transformants selected and recombinant plasmids extracted,the inserted gene was sequenced.Results\ The whole sequences of cyt b gene of swordtail fish were obtained.Conclusion\ Compared with sequences of cyt b genes in GenBank with BLAST,there were good homologies among swordtail fish and other fishes.The evolution tree built from the homology of cyt b genes of swordtail fish and other 13 kinds of fishes was consistent with the traditional taxonomic results.

Abstract:Objective To isolate ES cell colonies in 615 mouse. Method\ ES cell colonies in 615 mouse were isolated by PMEF feeder layer and the primary examination of these colonies without feeder layer was made. Results\ The formation rate of ES cell colonies and the rate of successful propagation were 22.6% and 0.94%,with positive ALP staining, stable karyotype and differentiation ability. Conclusion\ ES cell colonies in 615 mouse were isolated successfully.

Abstract:目的　研究中国版纳小型猪近交系动脉的异种靶抗原α -Gal的分布和半定量分析 ,为研究异种移植超急性排斥反应和异种生物材料提供资料。方法　通过亲和免疫组织化学法和图象分析对 10头版纳小型猪的四级动脉进行α Gal的分布和半定量研究。结果　 (1)各级血管组织中血管内皮细胞阳性表达明显 ,弹性纤维、胶原纤维和平滑肌细胞未见表达 ,外膜有微弱表达 ;(2 )图象分析显示 ,管径越细 ,其内皮细胞的表达越高 (P <0 .0 1) ,各级动脉内皮细胞阳性表达的面积密度比的组间差异显著 (P <0 .0 1) ,t检验发现每两组间的动脉内皮细胞阳性表达面积密度比相差显著 (P <0 .0 1)。结论　 (1)版纳小型猪动脉内皮细胞均有异种抗原α Gal分布 ,但是分布不均匀 ;(2 )微血管血栓是异种移植超急性排斥反应中的重要环节 ,小型猪的异种器官移植、彻底解决异种移植超急性排斥反应的关键是防治微血管栓塞

Abstract:Objective To study the overspeed method of cryopreservation of mice ovarian tissue, and find out a convenient and practical method of its storage. Methods After thawing, the ovarian tissues were transplanted. The recovered time and rate of estrus, level of serum estradiol of recipient mice, enzyme histochemistry of transplantation tissues, the ultrastructure of thawed tissues were studied to compare the cryopretective effect of cryopreservation methods and types of CPA. Results and conclusion In comparison with procedure freezing method,though the same CPA,the liquid nitrogen vapour freezing method showed the advantage of well ultrastructure of ovarian tissue in addition to the same results of enzyme histochemistry.There were no significant difference between the procedure freezing method and liquid nitrogen vapour freezing method in recovered rate of estrus, level of serum estradiol.

Abstract:Objective The cellular immunity and humoral immunity of the mice with gene knock out Smad3 were studied. Methods The parameters of CD4 ,CD3 ,CD8 ,CD19 ,IgG were detected by FCM and ELISA.Results The parameters of CD8 ,CD4 /CD8 ,IgG of homozygous mice were significant differences between genders. But the parameters of the CD4 ,CD8 ,CD3 ,CD19 ,IgG were lower than those of wild types. Conclusions It is reference value in using the mice lacking Smad3 for the study of disease of human immunity.

Abstract:Objective To make pathological observation of MMTV Wnt 1 transgenic mice with mammary tumor.Methods The mammary tumor of MMTV Wnt 1 transgenic mice was observed and the tumor tissue was transfered to nude mouse in site to check the pathological changes .Results Tumorgenesis occurred in spleen and liver while no obvious lesion appeared in other organs. No metastasis occurred at the time mammary tumors were detected.Transplanted tumors were with the similar biological features of primar tumor.Conclusion The MMTV Wnt 1 transgenie mice conld be an animal model with mammary tumor.

Abstract:目的　建立成年犬股骨微波灭活骨缺损的实验动物模型 ,为骨缺损修复的研究提供实验依据。方法应用自行研制的骨肿瘤微波治疗仪 ,以 1 5kHz频率、70W功率加热至 5 0～ 5 5℃ ,维持 2 0min ,造成犬股骨中段不同大小的骨缺损。结果　在保持犬股骨连续性的前提下 ,长度 1 5cm的骨缺损 9个月时有 1 2被新生骨填充 ,长度2 5cm和 3 5cm的骨缺损 9个月时无愈合倾向 ,但后者骨折的发生率高。结论　成年犬股骨微波高温造成的骨缺损 ,长 2 5cm、宽 1 0cm 9个月不能自愈 ,适宜于各种骨修复材料的填充 ,是理想的实验模型

Abstract:Objective To evaluate the feasibility of intrauterine fetal surgery in sheep. Methods Intrauterine surgery was performed at 116-125 gestational days on 4 ewes with twin fetuses each. Fetal toe or lip was excised in one of twin to observe the results of gestation and to evaluate histopathologic changes in newborn lambs. Results Up to full term gestation, all fetuses in the experiment were delivered through vagina and alive. The surfaces of the fetal wounds were without inflammation and scar formation. Histological examination of the skins and brains from both control and test groups demonstrated that all fetal lambs were normal except that only one brain tissue of the test group showed mild pathologic change. Conclusion Intrauterine fatal surgery is of feasibility.

Abstract:目的　建立检测犬细小病毒的PCR诊断试剂盒。方法　通过在犬细小病毒 (CPV)的基因组中设计的特异性寡核苷酸引物 ,利用PCR技术研制犬细小病毒的PCR诊断试剂盒。结果　在特定的反应条件下 ,使用该试剂盒能特异地扩增含有犬细小病毒的样品 ,且能检出痕量 (10ng)的犬细小病毒DNA ,被测粪样只需进行简单煮沸就能进行PCR反应 ,该试剂盒能在常温下保存 1周以上、4℃保存 1年以上。同血凝试验 (HA〕及单克隆抗体ELISA方法比较 ,对 6 6份样品进行检测 ,显示该PCR试剂盒具有特异、灵敏等优点。结论　成功研制了犬细小病毒的PCR诊断试剂盒 ,利用该试剂盒能从DNA水平上对犬细小病毒进行监控

Abstract:目的　构建pMal N重组表达载体 ,转化E .coliDH5α ,诱导表达犬瘟热病毒 (CDV)重组核衣壳 (N)蛋白。方法　采用RT PCR技术 ,从CDVRNA中扩增编码N蛋白的基因片段 ,通过连接反应 ,构建重组克隆载体和重组表达载体 ,转化感受态。E .coliDH5α细胞。通过IPTG诱导表达CDV重组N蛋白。结果　扩增出约 1 7kbCDV全长的主要结构蛋白N蛋白基因 ,通过PCR获得 5 36bpN蛋白基因片段。将N蛋白基因片段克隆入原核表达载体pMal C2 ,表达产物麦芽糖结合蛋白 (MAP)与N蛋白的融合蛋白的相对分子质量约 6 0× 10 3,与预期大小一致。结论　构建的pMal N重组载体所表达的CDVN蛋白为进一步研究CDV的特异、敏感的抗体检测方法打下基础

Abstract:目的　研究犬小肠缺血再灌注后氧自由基的改变、血管内皮细胞中的Bcl 2、Bax和血管内皮细胞生长因子 (VEGF)表达改变的意义及其相关性。方法　阻断分布于小肠较小范围的小肠动脉 ,建立小肠缺血再灌注模型。检测血中NO(Nu ml)和SOD(μmol L)浓度并以免疫组织化学方法对小肠组织中的Bcl 2、Bax和VEGF的表达及它们的相关性进行观察研究。结果　小肠缺血再灌注后 ,NO和SOD的浓度 ,Bcl 2、Bax和VEGF的表达有明显的改变。小肠再灌注 0min ,NO和SOD的浓度分别是 12和 75 ,Bcl 2、Bax和VEGF的阳性细胞率分别为 85 % ,5 5 %和10 % ,再灌注 30min ,NO和SOD的浓度达最底 (11和 4 3) ,Bcl 2、Bax和VEGF分别为 6 6 %、5 4 %和 6 5 % ,而再灌注 6 0min ,NO和SOD的浓度恢复至 15和 90 ,三种基因的表达则分别为 4 4 %、75 %和 5 %。在对照组仅有少量阳性细胞。结论　小肠缺血再灌注可引起NO和SOD浓度降低、Bcl 2和VEGF表达增加 ,但随着血流恢复NO和SOD的浓度有所回升而Bcl 2和VEGF阳性细胞逐渐减少。而凋亡基因Bax的表达则逐渐增加。小肠缺血再灌注能引起氧自由基的增多、凋亡基因表达增强且有明显的相关性

Abstract:Objective To establish the model of diabetic retinopathy complication in rhesus monkey. Methods Four adult, healthy and male rhesus monkeys were injected intravenously with different STZ dosage(one of the four with 60 mg/kg, one with 45 mg/kg, the other two with 30 mg/kg twice fifteen days apart). Result All of the monkeys were made animal models of the insulin dependent diabetes mellitus(IDDM) and the state of chronic hyperglycemia(SCH). These diabetic monkeys had long term higher preprandial plasma glucose, and they showed tiny blood vessel dilation in eyeground, early retinal hemorrhage dot, tiny angioma and cataract, during 63-91days after inducing STZ. Conclusion Animal model of diabetic retinopathy complication disease was established. It would be useful for studying the mechanism of diabetes mellitus and its complication, and observing the efficiency, safety of the diabetic drugs instead of human being.

Abstract:Objective Structure of myocardial mitochondria between FMMU albino guinea pigs and pigment ones was compared after exposed to hypoxia. Methods FMMU albino guinea pigs and pigment ones were placed in hypoxic chamber at normal pressure for 7 d,14 d,21 d respectively to make hypoxic model. Cardiac muscle was taken out and the structure of myocardial mitochondrial was observed under electron microscope. Results In the same hypoxia the myocardial ultrastructure of FMMU albino guinea pigs and pigment ones changed in varying degrees. In different hypoxic stage the ultrastructure of the same guinea pigs changed in varying degrees, too. Conclusion Change degrees of myocardial ultrastructure of FMMU albino guinea pigs were lighter than that of pigment ones under hypoxic circumstances. It hinted that the anoxia tolerance of FMMU albino guinea pigs was higher than that of pigment ones.

Abstract:Objective To compare protein expression of EAC stored in different institutes and its cloned cells. Methods According to Fdler and Kripke's experimental evidence and theory, 5 cloned cells were isolated from EAC in Beijing Cancer Institute by single cell cloning technique. Proteins of EAC from Wuhan, Shandong,Beijing Cancer Institute and its cloned celles were compared by SDS PAGE and immunohistochemistry staining. Results The number of protein bands of EAC in Wuhan, Shandong and Beijing Cancer Institute was 22,25 and 28 respectively. The number of protein bands of cloned cell E2G8 was 26. The EAC cells in three institutes had different in the reaction to 5 kinds of antibodies and 5 cloned cells had different reactions to 10 kinds of antibodies. Cloned cell E2G8, E2F4 had positive reaction to 7 antibodies,while E2C6 to 8 antibodies and E1G5,E2B5 to 6 antibodies. And the positive reaction rates were all beyond 85%. Conclusion There were remarkable difference among EAC from different institutes and its cloned cells.

Abstract:Objective To study the effects of herbicide LeCaoLong on DNA damage of red crucian carp lymphocyte in vivo . Method Single cell gel electrophoresis (SCGE) assay was used to detect DNA single strands breaks (DNA SSBs) when red crucian carps were exposed to different concentration of LeCaoLong. Results At the concentration of 0 88, 1 75, 3 50, 7 00 mg/L, LeCaoLong could induce DNA damage in the peripheral blood lymphocyte of red curcian carp, the comet cell tail length of every group exposed to LeCaoLong was longer than the control group( P<0 05 ). The DNA damage was signficantly positively correlated with concentrations in the range of 0-7 0 mg/L,( r=0 982,P<0 01 ), when red crucian carps exposed to LeCaoLong for 12 h,24 h,48 h,96 h and 10days. The comet cell rates of every concentration group were higher than relative controls. Conclusion All the results suggest that the herbicide can cause significant genetic damage to the lymphocyte of red crucian carp at certain concentration and time.

Abstract:The technique route for making transgenic animals by somatic cell mediated transgene has been developed based on the technique of nuclear transfer. Although it came into being for only about 4-5years, it has shown broad application prospect and great potential. In this review, we compared advantages and disadvantages of different methods for making transgenic animals, expounded the basic technique route for making transgenic animals by somatic cell mediated transgene, emphasized the key points of somatic cell transfection and cloning of integrated cells, and the limitation and potential application of this technique.