Abstract:目的 构建HLA-DRα和HLA-DRB1*0405真核表达载体,并使其在哺乳动物细胞内得到表达.方法 从含HLA-DRα全长cDNA的质粒中扩增HLA-DRα的ORF序列,通过酶切将载体pIRES2-EGFP中的EGFP切除,将HLA-DRα ORF插入到载体中;采用RT-PCR方法 从HLA-DRB1*0405阳性人外周血单个核细胞中克隆出HLA-DRB1*0405 ORF序列,将其插入pIRES2-DRα载体的多克隆位点处,构建出真核表达载体pIRES2-DRαβ1*0405.对构建的载体进行限制性内切酶鉴定和测序.采用脂质体转染方法 将表达载体转入小鼠成纤维细胞系DAP2.3中,通过流式细胞术和间接免疫荧光法对HLA-DRα和HLA-DRB1*0405的表达进行检测.结果 酶切鉴定和测序证实目的 基因插入正确且序列与GenBank一致.流式细胞术检测约41.41%的转pIRES2-HLA-DRαβ1*0405载体的细胞呈阳性,间接免疫荧光显示HLA-DRα和HLA-DRB1*0405在转基因细胞中得到表达并形成完整的HLA-DR4分子,表达于胞膜及胞质内.结论 成功构建出真核表达载体pIRES2-DRαβ1*0405,并在小鼠成纤维细胞系中得到表达,为下一步进行稳定细胞系的建立和转基因鼠模型的构建奠定了基础.

Abstract:目的：构建SDF-1α真核表达载体，为研究SDF-1α基因功能提供工具。方法：从大鼠平滑肌细胞中提取总RNA，采用RT-PCR技术扩增SDF-1α的基因编码区序列，将序列定向克隆至真核表达载体pcDNA3．1上，并用PCR、酶切法和DNA测序鉴定。结果：PCR和限制性内切酶分析鉴定，表明获得重组质粒pcDNA3．1／SDF-1α。通过对SDF-1α基因编码区cDNA序列测序证实其与GenBank中的已知序列一致。结论：成功构建了SDF-1α真核表达载体，为SDF-1α基因功能研究奠定基础。

Abstract:Objective To identify the effectiveness of McAb(cM-T807) anti-huCD8~+T cell on eliminating the CD8~+ lymphocytes from Chinese original rhesus monkeys.Methods The anti-CD8~+T cell McAb was injected into two rhesus monkeys at 0 d,3 d and 7 d,respectively.Peripheral blood and lymph node samples were collected from the two monkeys at different time after injection.The CD3/CD4/CD8 from both peripheral blood lymphocytes(PBL)and lymph nodes(LN) cell suspension were differentiated by FACS and the effectiveness of elimination of CD8~+ lymphocyte was detected.Results The results showed that 99.9% and 90.0% reduction of CD8~+cells in peripheral blood and lymph nodes from two Chinese-origin rhesus macaques were detected from 24 and 168 hours after injection,respectively,and the duration of low CD8~+ levels maintained for 27 days in PBL and in LN for 21 days.Conclusions CD8~+T cells can be eliminated effectively from PBL and LN in Chinese original rhesus monkeys by injection of McAb cM-T807.

Abstract:目的：测试用抗-HuCD8^＋T细胞单克隆抗体方法去除中国恒河猴体内CD8^＋T细胞效果，为建立CD8^＋T细胞缺失恒河猴模型提供实验依据。方法：抗-HuCD8^＋T抗体cM-T807在0、3、7d注射2只中国恒河猴，不同的时间取外周血，腹股沟和腋下淋巴结。制备CD3／CD4／CD8标记的外周血和淋巴细胞悬液，流式法测定恒河猴外周血和淋巴结中CD8^＋T细胞的去除效果。结果：注射了cM-T807的中国恒河猴，24h后，外周血中的CD8^＋T细胞缺失99．9％，持续27d；168h后，淋巴结中CD8^＋T细胞缺失约90．0％，持续21d。结论：首次证实抗-HuCD8^＋T细胞抗体cM-T807能有效去除中国恒河猴体内CD8^＋T细胞，为建立CD8^＋T细胞缺失动物模型（CD8^＋T Cells del）奠定基础。

Abstract:Objeelive To provide normal physiological and hematological indexes ofWacaca fascicularis. Methods 12 male and 12 femaleWacaca fasc/cu/ar/swere chosen for this study. Results T, GLU, Acidophilic normoblast, the ratio of leukocytes to rhodoeytes, body weight, lung and thymus showed significant difference between both sexes. Other indexes were not significantly different.

Abstract:Objective To evaluate the feasibility of Bama miniture pigs to be used as a model on pharmacokenetic properties of CYP3A by comparing enzyme kinetics of lovastatin metabolism in liver microsomes of Bama miniture pigs, humans and rats. Methods Liver microsomes were prepared from Bama miniature pigs, humans and rats, respectively. To evaluate the biotransformation activity in the minipig liver microsomes and compare with those of humans and rats, incubation with lovastatin was carried out. The inhibitory effects were observed by using chemical inhibitor specific to the CYP3A in humans. Results The enzyme kinetics in liver microsomes of minipigs were more similar to humans than rats. The metabolism of lovastatin in three various genera was significantly inhibited by ketoconazole. Conclusion Compared with rats, Bama miniature pigs are more suitable for the pharmacological studies related to CYP3A.

Abstract:目的：探讨黄芪多糖对小鼠1-细胞胚胎体外发育的作用。方法：以mCZB添加抗生素为对照组，以添加不同浓度黄芪多糖（Astragulus Polysacharin，Aps）为试验组，观察胚胎体外发育率、孵化胚胎细胞数目及切割取样后桑椹胚发育率和细胞数目。结果：体外培养96h，试验组囊胚发育率（86．0％、85．8％、89．5％）均显著高于对照组（75．8％）（P〈0．05），各试验组间差异无显著性（P〉0．05）；体外培养144h，试验组胚胎孵化率（28．9％、31．0％、25．4％）均高于对照组（22．6％），以Aps2（100μg／mL）组孵化率为最高（31．0％）。孵化胚胎细胞数试验各组（76．00±6．25、81．73±7．01、75．89±4．54）均极显著高于对照组（69．06±5．15）（P〈0．01）。切割取样（≤5或〉5个）后桑椹胚体外培养72h，试验组发育率和细胞数分别为（≤5组为50％和44±2．7、50％和53±2．7、44．4％和55±2．7；〉5组为23．1％和42±1．6、20．0％和44±1．0、14．3％和41±1．0）均高于对照组（33．3％和41±1．7；0％和0）。结论：中药有效成分黄芪多糖能促进小鼠早期胚胎的体外发育及胚胎细胞增殖，并对切割取样后胚胎的体外培养有一定促进作用，且切割样本数≤5对胚胎发育比较有利。

Abstract:Objective To establish an effective method for superovulatory treatment of rats with pregnant mare serum gonadotrophin(PMSG) and human chorionic gonadotropin(hCG). Methods Sprague-Dawley rats at different age(4-5 weeks,67 weeks and 8-9 weeks) were superovulated with PMSG and hCG at a dosage of 10 IU, 20 IU,30 IU,40 IU or 50 IU,respectively.The results of superovulation of those rats were analyzed.Finally,the 2-cell embryos fertilized in vivo were transferred to recipients to observe the rate of pregnancy.Results Rats aged 4-5 weeks showed the best effect of PMSG/hCG at a dosage of 20 IU on superovulation,with an average number of 54.6 per rat,while rats aged 6-7 weeks and 8-9 weeks were 32.7 and 21.5 per rat,respectively.After embryo transfer,the rate of pregnancy was 63.3%,showing no significant difference with that of control rats(60%).Conclusion Rats at 4-5 weeks of age are more sensitive to superovulation,giving a highest number of oocytes with 20 IU PMSG and hCG.Such oocytes obtained by superovulation are good in quality and can complete the whole term development after fertilized in vivo and embryo transfer.

Abstract:目的：探讨丙泊酚的遗传毒性及对遗传物质可能产生的损伤作用。方法：应用鼠伤寒沙门氏菌回复突变实验、单细胞凝胶电泳实验及小鼠骨髓微核实验，测丙泊酚对基因突变、染色体改变和原发性DNA损伤3个遗传学终点的影响。结果：丙泊酚在每皿1．0mg～0．01mg范围内，有或无代谢活化系统时，TA97、TA98、TA100和TA1024种菌株的每皿平均回变菌落数未见阳性的剂量-反应关系。单细胞凝胶电泳实验：小鼠经丙泊酚1．5mg／kg、3．0mg／kg、6．0mg／kg和12．0mg／kg染毒后外周血淋巴细胞彗星率分别为4％，6％，5％和6％；彗星尾长分别为：（21．51±0.48）,μm、（21．67±0．58）,μm、（21．89±0.48）,μm、（22．01±0.66）,μm。各组彗星率、彗星尾长与阴性对照组比较差异无显著性（P〉0．05）。骨髓微核实验：4个剂量组的微核发生率分别为（2．87±0．64）‰、（3．0±0.53）‰、（3．12±0.64）‰和（3．25±0.89）‰，各组微核率与阴性对照组微核率（2．50±0.53）‰比较差异无显著性（P〉0.05）。结论：在3个配套遗传学终点实验中未见丙泊酚有遗传毒性作用。

Abstract:Objective To evaluate the safety of cornstalk bedding material for SD rat breeding.Methods All SD rats were randomly divided into control group and cornstalk groups.The rats were killed after being raised for 6 months.The blood biochemical indexes were measured,and histopathological examination of a number of organs were performed.Results The eight blood biochemical indexes of SD rats in the two groups were all in normal range and showed no significant difference(P>0.05).No significant pathological changes were observed in the examined organs.Conclusion The cornstalk bedding material can be safely used in SD rat breeding.

Abstract:目的：不同剂量氯胺酮麻醉大鼠后，进行麻醉效果评价及生理指标观察。方法：3组不同剂量氯胺酮（100、120、150mg／kg）分别腹腔注射大鼠，进行大鼠左肾切除术。记录麻醉起效时间、麻醉维持时间；无创心电监护仪监测大鼠心率（HR）、呼吸（RR）、氧饱和度（SO2）、体温（T）；进行镇痛评价、呼吸道评价、肌肉松弛度评价。结果：3组剂量中，以氯胺酮120mg／kg体重腹腔注射大鼠，麻醉持续时闯45±9．59min，HR、RR波动小，镇痛效果、肌肉松弛度较好；以氯胺酮100mg／kg体重腹腔注射大鼠，麻醉效果较差，麻醉时间在27．0±3．033min，HR、RR、S02指标波动大；以氯胺酮150mg／kg注射大鼠，麻醉效果虽好，但呼吸受抑制，体温较另外两组低。结论：以氯胺酮120mg／kg体重腹腔注射大鼠，麻醉效果评价较其他两组好，可满足于45min左右的大鼠实验。

Abstract:Objective To assess the toxic effect of 2% peracetic acid aerosol disinfection on laboratory animals and the staff. Method Isolator was disinfected with 2% peracetic acid aerosol at concentration of 1 g/m^3 density for 4 hours. Rats were kept in the isolator and their behavior and pathological changes were observed to assess whether inspiration of the aerosol lead to toxicity in the rats. Results After 2 % peracetic acid aerosol inspiration for 4 hours, the rats depicted abnormal behavior activity during 4 hours to 7 days. Conclusions 2% peracetic acid aerosolat using a density of 1 g/m^3 is toxic to laboratory animals.

Abstract:Objective To report the experience in establishing a swine model of acute myocardial infarction(AMI) by coronary occlusion with balloon angioplasty.Methods Fourteen pigs aged 3-5 months and weighed 25 kg-40 kg were anesthetized.Balloon angioplasty was positioned in the left anterior descending(LAD) coronary artery at the beginning of the second septal branch.The artery was blocked for 1 min,2 min,5 min by turns,with an interval of 60 seconds every time,as a way of ischemic preconditioning in group A(n=7).Then the balloon was inflated at 2 atm to occlude the LAD for 60 min.The balloon was inflated at 2 atm to occluded the LAD for 60 min without ischemic preconditioning in group B(n=7).ECG and blood pressure were monitored.The ischemic change was comfirmed by MRI.Results Twelve animals were set up the acute myocardial infarction model successfully.Two died of ventricular fibrillation in group B.During operation,twelve pigs generated ventricular fibrillation,among them 10 were salvaged successfully.Ten pigs were confirmed as anterior septal AMI and two was confirmed as widespread anterior AMI by ECG.MRI comfirmed AMI in tweleve pigs.Conclusion Swine model of acute myocardial infarction can be estattished successfully by coronary occlusion with balloon angioplasty.Ischemic preconditioning is necessary to prevent ventricular fibrillation.

Abstract:目的：探索一种复制小鼠系膜增生型。肾小球。肾炎动物模型的方法。方法：80只C57BL／6小鼠随机分为4组（每组20只）：A-正常对照组；B-竹叶青蛇毒2mg／kg注射组；C-竹叶青蛇毒4mg／kg注射组；D-竹叶青蛇毒6mg／ks注射组。按照分组要求计算B、C、D3组小鼠中每只小鼠需注射的竹叶青蛇毒的总量，配制3种不同浓度的竹叶青蛇毒液，B、C、D3组小鼠分别尾静脉注射不同浓度的竹叶青蛇毒液0．2mL，A组小鼠尾静脉注射生理盐水0．2mL以进行对照。结果：竹叶青蛇毒注射后的小鼠均出现中毒性症状。C、D组小鼠因蛇毒注射剂量较大，死亡率高达50％，且集中于前3d内死亡，3d后中毒小鼠的活动基本如常。B组与正常对照组小鼠无死亡。中毒小鼠的肾功能与正常对照组相较无明显差别，但均出现了肾小球系膜细胞增生，系膜基质增多，肾小球及间质内炎性细胞浸润等病理学改变。结论：竹叶青蛇毒尾静脉注射是一种复制小鼠系膜增生型肾小球肾炎动物模型的较好的方法。

Abstract:Sleep apnea syndrome(SAS) causes various diseases and especially impacts cardiovascular system.Currently,different animal models related to cardiovascular diseases were established with varying pathophysiological state of human beings.However,none of them was supposed to be perfect.Along with the researches,SAS animal models are being well developed.

Abstract:Objective In addition to a fundamental role in very diverse processes such as vision and growth and differentiation of numerous types of cells,vitamin A(retinol) is involved in the regulation of testicular functions in rats and mice.An excess of vitamin A leads to spermatogenetic disorders,and its deficiency induces early cessation of spermatogenesis.Furthermore,mouse mutant are steriles.In this review,the location of retinoic acid receptors,the metabolisms of action in the testis and its effects on fetal and adult testis are summarized.