Abstract:Objective To analyze the genetic purity of swordtail fish (Xiphophorus helleri) inbred strain for directing the breeding of swordtail as laboratory animal. Methods 29 selected single primers were used to amplify randomly genomic DNA of the inbred strain of swordtail fish. The amplified fragment bands were analyzed to calculate the similarity index and the genetic distance. Results 306 bands with the lengths between 200bp and 3kb were obtained, among them there were 29 polymorphic bands, the frequency of polymorphic bands was between 0-50.00% and the average was 9.48%. There was a high frequency of common band in different individuals. Based on RAPD patterns, the calculated similarity index within 12 samples of the inbred strain was 0.9839(0.973-0.997), the mean allelic frequency was 0.8731, and the minimum heterozygosites was 0.1269. Conclusion The inbred strain of swordtail fish has reached a high degree of genetic purity.

Abstract:Objective In order to obtain a model of the HEK293 cells infected by porcine endogenous retroviruses PERV, by means of the G418|resistant HEK293 cell line established. Methods The plasmid pIRESneo containing neo gene was purified and transfected into HEK293 cells with Lipofectin. The transfected cells would be survived in the further culture medium containing G418 antibiotic as the neo gene could express a G418 resistant products. The identification of the G418 resistant HEK293 (G418 RHEK293) cells was conducted by polymerase chain reaction with the specific primers of neo gene. Results The G418 RHEK293 cell line was established successfully. There were no differences among G418 RHEK293 cells and normal HEK293 cells morphologically and propagatively. The neo gene DNA fragment could be amplified by PCR with specific primers in the genomic DNA of G418 RHEK293 cells. Conclusion The G418 RHEK293 cells were established with transfection of plasmid pIRESneo by Lipfectin, which may be useful as a cell model for the research of PERV.

Abstract:Objective To develop a specific RNase Protection Assay to measure black seabream growth hormone (GH) receptor mRNA. Methods GH receptor cDNA fragment of black seabream was subcloned in pGEM|T vector. Then a specific antisense GH receptor RNA probe with radioactivity was prepared to hybridize with sense GH receptor RNA and total RNA from tissues of black seabream respectively. After hybridized solutions were treated with RnaseA/RNAseT1, the protected double strand hybridized RNAs could be detected by Autoradiogram and radioactivity counting. Results A specific RNase Protection Assay to measure black seabream GH receptor mRNA was developed. GH receptor mRNA were detected in brain, liver, kidney, muscle, gonad, gut and gill of black seabream, with highest level in liver. The results of GH receptor mRNA distribution were accorded with our previous results by GH Radioreceptor Assay in black seabream. Conclusion We can study the molecular mechanism of endocrine regulation by the specific RNase Protection Assay in black seabream GH receptor.

Abstract:Objective To study the activities of the seroenzyme of the Smad3 gene knock|out mice.Methods The parameters of ALP,AST,ALT,CK,LDH|L of the 35 days,70 days and 6 months mice were measured by VITALAB made in Holland.Results The parameters of the homozygous mutation mice were the highest among that of the three gene types.ALP decreased with age,but AST,ALT,CK and LDH|L increased with age.Conclusion The activities of the seroenzymes changed due to lacking of the gene.

Abstract:目的　探讨小鼠卵母细胞孤雌激活的最佳作用条件。方法　将MⅡ期小鼠卵母细胞随机分为 2组 ,第一组 ,将小鼠卵母细胞分别放入 2、4、6、8、10mmol ml不同浓度的氯化锶激活液中 ,作用 6h ,观察激活率及囊胚发育率。第二组 ,将小鼠卵母细胞放入 10mmol ml氯化锶激活液中 ,分别作用 3、6、9h ,观察激活率及囊胚发育率。结果　第一组 ,激活率分别为 86 49%、82 6 1%、88 0 0 %、86 6 7%、81 18%。各组间差异无显著性。体内培养 72h回收囊胚 ,囊胚发育率分别为 0、31 42 %、43 33%、6 2 5 0 %、5 0 0 0 %。 6～ 10mmol mlSrCl2 激活液激活后囊胚发育率高于 0～ 4mmol ml(P <0 0 5 )。第二组 ,激活率分别为 82 86 %、89 6 1%、91.40 %。 72h囊胚发育率分别为 2 6 5 3 %、5 0 0 0 %、5 3 2 2 %。激活 6、9h的囊胚发育率高于激活 3h的囊胚发育率 (P <0 0 1)。结论　结果表明 ,6～ 10mmol ml的SrCl2 为卵母细胞孤雌活化的最佳作用浓度 ,6～ 9h的激活时间为最佳作用时间 ;表明SrCl2 的浓度和作用时间对小鼠卵母细胞的活化有显著的影响。

Abstract:目的　构建人胰岛素基因真核高效表达载体 ,为胰岛素转基因研究奠定基础。方法　用限制性内切酶SpeⅠ和HindⅢ消化pEF1α GFP ,回收 1 2kbEF1α启动子 ,插入到pCMV mINS的NruⅠ和HindⅢ位点中 ,获得重组质粒pEF1α mINS ;将pCMV mINS和pEF1α mINS分别转染BHK细胞 ,用G418筛选 ,阳性克隆传至 2 0代后 ,分别用放免方法和免疫组化法分析胰岛素和 或胰岛素原在BHK细胞中的表达情况。结果　经放免测定 ,pCMV mINS和pEF1α mINS在BHK细胞中胰岛素和 或胰岛素原的表达量分别为 4 0 77μIU ml和 6 897μIU ml。经免疫组化分析 ,pCMV mINS在BHK细胞质中胰岛素表达水平的灰度值为 190 0± 19 5 6 ;pEF1α mINS在BHK细胞质中胰岛素表达水平的灰度值为 181 4± 18 45 ,在BHK细胞核中表达水平的灰度值为 15 5 4± 11 6 6。结论　在BHK细胞中启动子EF1α启动胰岛素基因表达的活性比启动子CMV高。

Abstract:Objective The mutant gene of BALB/c hairless mice affects skin and coat structure, but whether can it affect the immune function. Methods the parameters of CD4 +,CD3 +,CD8 +,CD19 +,IgG were tested by FCM and ELISA.Results 1.There were no significant differences in mice between sexes. 2. The parameters of the hairless mice were lower than those of other phenotypes. The parameter of CD8 + was significantly different by statistics. There were no significant differences among IgG of three phenotypes.Conclusion The mutant gene of BALB/c hairless mice affected the immune function.

Abstract:目的　建立实验大小鼠金黄色葡萄球菌的快速检测方法———PCR法。方法　根据已公布的金黄色葡萄球菌耐热核酸酶nuc基因的序列 ,设计并合成一对特异性的引物 ,利用PCR技术扩增nuc基因片段。对金黄色葡萄球菌和其他非金黄色葡萄球菌菌株抽提的DNA进行扩增。结果　金黄色葡萄球菌PCR产物出现 6 6 8bp的特异性DNA扩增片段 ,而其他非金黄色葡萄球菌未出现扩增片段 ,证实了合成的引物对金黄色葡萄球菌具有特异性。将抽提的金黄色葡萄球菌DNA进行系列稀释 ,测定此PCR体系的敏感性 ,结果显示 ,该PCR体系能检出 3pg金黄色葡萄球菌DNA ,且从抽提DNA到PCR扩增及电泳结束仅需 4h。结论　本研究所建立的扩增耐热核酸酶nuc基因检测鼠金黄色葡萄球菌的PCR方法 ,具有快速、可靠、敏感和特异的特点 ,可用于临床样品和金黄色葡萄球菌感染时的检测 ,适合应用于实验大小鼠的监测。

Abstract:目的　研究链脲菌素 (STZ)处理新生期大鼠后诱发成年发病糖尿病的量效关系及其胸腺淋巴细胞增殖活性的变化。方法　STZ分别以 2 0、40、6 0及 10 0mg kg ,ip ,给予Wistar新生期大鼠 ,于注射STZ后第 3天、第 7天、第 17天、第 42天观察体重、血糖、血浆胰岛素及胸腺淋巴细胞增殖反应的动态变化。结果　 10 0mg kg剂量组可于第 42天形成慢性糖尿病模型 ,血糖值 (16 8± 4 6 )mmol Lvs (5 8± 1 6 )mmol L ,P <0 0 0 1;血浆胰岛素水平(5 2± 1 2 )mIU Lvs(8 4± 1 6 )mIU L ,P <0 0 1) ,胸腺淋巴细胞增殖呈现升高、受抑制再恢复的变化过程。其他各剂量组 ,特别是 40mg kg虽无明显高血糖形成 ,但可使胸腺淋巴细胞增殖活性增强。结论　STZ(10 0mg kg,ip)处理新生期大鼠诱发糖尿病的最适剂量为 10 0mg kg ,诱发时间为 42d。诱发过程中伴有胸腺增殖活性的变化。

Abstract:目的　研究慢性束缚应激时大鼠行为学和免疫功能的变化以及疏肝、健脾、补肾三种中药复方对其影响。方法　用特制束缚架连续束缚 2 1d(或 7d) ,每天 3h的方法制作大鼠束缚应激模型 ,观察大鼠行为学和免疫功能的变化。结果　慢性束缚应激后 ,大鼠的行为学发生了明显的变化 ,血清IL - 1β 上升 ,IL - 2和IL - 6下降 ,并且随着束缚时间的延长变化越明显。结论　逍遥散、四君子汤和金匮肾气丸能改善大鼠行为学的变化 ,明显增强慢性束缚应激大鼠的免疫功能。

Abstract:Objective The effects of glucose, pyruvate and lactate on in vitro development of bovine embryos produced in vitro in chemical defined culture system (SOF+PVA) were observed in this research. Method Bovine oocytes were matured and fertilized in vitro, then the fertilized ova were cultured in vitro in chemical defined culture medium. Results Experiment 1: Bovine IVF embryos were cultured in SOF+PVA that was not supplemented with glucose, then were cultured in SOF+PVA supplemented with 0, 1.50, 3.30 and 5.00mmol/L glucose respectively after being cultured 120 h, the embryos in control group were cultured in SOF+PVA supplemented with 1.50 mmol/L glucose during all the culture time. The results showed that the development rates of blastocysts were 9.2% a, 12.1%, 19.2% b, 18.9% and 11.7% respectively (a

Abstract:Objective To study the genetic variation and inbreeding of Xishuangbanna miniature pig.Methods The polymorphism of 12 microsatellites loci in the Xishuangbanna miniature pig inbreed(XMI) was analysed. Results The dendrogram results based on genetic distance suggested no difference between XMI and their pedigree, and that rates of homologous genes in them were high. Conclusion XMI has many substrains with different characteristics and high inbreeding degree.

Abstract:Objective To synchronize the estrus of female mouse model for avoidance of being conceived by mating. Mothods Levonorgestrel and quinestrol tablets were used to establish the female mouse model of synchronized estrus, with diethylstilbestrol as control. Results With dosage of levonorgestrel and quinestrol tablets administrated for mouse twice as person's once every five days, the probability of synchronized estrus of female mouse would come to or exceed 80%. Conclusion This experiment provided simple and convenient method for establishment of Kidney Yang Deficiency due to !excess of sexual intercourse".;

Abstract:Objective For the purpose of studing superovulation in C 57BL/6J mice. Method 5IU+5IU or 10IU+10IU PMSG and HCG were used to stimulate superovulation of C 57BL/6J mice. After PMSG given at 4:30 p.m. on the first day, HCG was injected on the third or fourth day (48 h or 72 h later). Ova were flushed out of the oviduct 16h after administration of HCG. The ova's amount of each group was compared. Result The superovulation in first group (5IU+5IU PMSG and HCG, 48 h) was best. The ova's amount of mice in 8-10 weeks by superovulation was more than that in 6-8 weeks. Conclusion In C 57BL/6J mice, the best dosage of hormone for superovulation is 5IU PMSG + 5IU HCG, and the best interval of time for the administration is 48h.

Abstract:Objective To study physiological data of food and water intake, urination, body weight, plasma glucose and glycosuria with streptozotocin(STZ) in rhesus monkeys, which was essential to establish animal model for diabetes mellitus(DM). Methods Seven rhesus monkeys were injected intravenously with different doses of STZ. Results The seven monkeys showed the same clinical features as those of diabetes mellitus patients at a different time and extent, such as polyphagia, polydipsia, polyuria, hyperglycemia, hyperglycosuria, body weight loss. The clinical characteristics were lightened and controlled by using insulin. Especially the clinical symptoms of mid-dose and high-dose of STZ were more obvious than those of low-dose in monkeys. The body weight of low-dose monkeys rose in a short time, then dropped rapidly. Plasma glucose and glycosuria of monkeys were increased with STZ injected, especially mid-dose and high--dose monkeys varied obviously. Conclusion The acute animal model similar to human DM was induced by mid-dose and high-dose of STZ injected. The course of DM animal model may be extended by using insulin treatment or low-dose of STZ, which is suitable for studying complications of DM.

Abstract:Objective To observe the effect of LINGGUANG injection(compound anisodine)on heart and liver after hemorrhagic shock in rats. Method Fifty-six male Wistar rats were divided into 4 groups randomly, which were sham shock, shock, shock treated with LINGGUANG injection in low dose and high dose groups separately. Animals were subjected to sham shock, or 70 minutes shock at 4-kPa blood pressure with or without LINGGUANG injection treatment. Half of the rats were sacrificed at 6 hours after resuscitation, the rest were killed at 12 hours. The CK,CK-MB,LDH,ALT,AST of sera were detected. The morphological changes of hearts and livers were observed under light microscope and transmission electron microscope. Results and Conclusion Compound anisodine is effective for curing the damages of heart and liver caused by hemorrhagic shock and reperfusion. The mechanism of compound anisodine treatment may be related with microcirculation improving, free radical scavenging.