Abstract:Objective To construct bioreactor with housefly to express human insulin.Methods Four recombinant expression plasmids were transferred respectively into the new fertillized eggs of housefly by electroporation.The first generation was screened with G418 after the founders were incubated to adults.Human insulin was detected by radioimmumoassay, Tricine-SDS-PAGE and Western-blot in samples of maggot of the second generation collected at the time of 96?hours.Results All four recombinant expression plasmids could express human insulin in housefly. Conclusion The housefly maggot has the ability of processing post-translation insulin genomic gene.So housefly can be used as bioreactor to produce human insulin.

Abstract:目的 利用PCR—RFLP和PCR—SSCP技术分别检测犬MC1R基因中两个序列T105A和R306Ter的基因多态性，为遗传育种、培育出更加优良的实验用犬奠定基础。方法 以224只犬为实验材料，对犬MC1R基因T105A片段和R306Ter片段分别进行PCR-RFLP和PCR—SSCP分析，并且分别对具有RFLP和SSCP多态性的片段进行克隆测序，找出等位基因的差异位点。结果 经对犬MClR基因的PCR-RFLP分析，发现犬MC1R基因的T105A序列具有多态性，表现为三种基因型：AA、AB和BB。通过对具有RFLP多态性的片段进行克隆测序发现MC1R基因在编码第105位氨基酸的密码子处存在由G到A的单碱基突变，该突变导致第105位氨基酸发生由精氨酸向苏氨酸的改变，此外在第207位氨基酸的密码子处还发现由A到G的单碱基突变，并产生了一个HhαI限制性内切酶位点。同时，经过犬MC1R基因的PCR—SSCP分析，发现犬MC1R基因的R306Ter序列具有多态性，表现为三种基因型：CC、CD、DD.通过对具有SSCP多态性的片段进行了克隆测序发现，MC1R基因在编码第306位氨基酸的密码子处存在一个由CGA到TGA的终止突变，而使翻译终止。结论 利用PCR—RFLP和PCR—SSCP分析技术证实犬MC1R基因具有明显的多态性。

Abstract:目的　分析传染性喉气管炎病毒 (ILTV)gC蛋白的免疫活性。方法　设计引物扩增编码gC蛋白主要免疫原性区域cDNA ,将其克隆到原核表达载体pGEX_6P_1的EcoRⅠ和SalⅠ位点之间 ,构建重组质粒r_pGEX_gC。将r_pGEX_gC转化到大肠埃希菌BL2 1(DE3)株中。结果　经IPTG诱导后 ,GST gC融合蛋白在重组菌株获得表达 ,表达产物相对分子质量为 6 5× 10 3,与预计相对分子质量大小一致。融合蛋白的表达含量占整个菌体含量的 2 7 8%。West ern_blot结果表明gC蛋白具有良好的反应活性。结论　本研究为传染性喉气管炎诊断抗原研制奠定基础。

Abstract:含有产气荚膜梭菌α毒素基因序列的克隆载体pMD18TCα ,经BamHⅠ和HindⅢ内切酶双酶切后 ,将α毒素亚单位基因亚克隆到原核表达载体pET30b中 ,构建表达载体pET30bPα。经酶切和测序鉴定 ,α毒素基因中的90 0bp的片段正确插入到表达载体中。重组菌株BL2 1(DE3)pET30bPα经IPTG诱导的表达产物经SDS PAGE和Western_blot分析 ,表达出大小为 38× 10 3的重组蛋白具有和α毒素阳性血清发生特异性的免疫反应。

Abstract:目的　观察松木、玉米秸、麦秸 3种垫料物质对大鼠血液生化指标和肝脏细胞色素P4 50 (CytP4 50 )酶系的影响。方法　将SD大鼠随机分为松木组、玉米秸组、麦秸组和对照组 ,于饲养 1、2、3、4周时测定血液中的丙氨酸转氨酶 (GPT)、白蛋白 (ALB)和总蛋白 (TP) ,测定肝脏细胞色素P4 50 酶 (CytP4 50 )、细胞色素b5酶 (Cytb5)和NADPH_细胞色素C还原酶。结果　 3个实验组与对照组的GPT、ALB和TP值相比无统计学差异 (P >0 0 5 ) ,且均在正常范围之内。松木在 2～ 4周对CytP4 50 有显著诱导作用 (P <0 0 5 ) ,第 3周差异极显著 (P <0 0 1) ;在 1～ 4周对NADPH_细胞色素C还原酶有诱导作用 (P <0 0 5 ) ,其中 2～ 4周差异极显著 (P <0 0 1) ;在 2、3周对Cytb5有诱导作用 (P <0 0 5 )。麦秸在第 2、3周可以诱导CytP4 50 和Cytb5的活性增加 (P <0 0 5 ) ,在 3、4周可以诱导NADPH_细胞色素C还原酶的活性增加 (P <0 0 5 ) ;玉米秸在 2、3周时对CytP4 50 和NADPH_细胞色素C还原酶均有诱导作用 (P <0 0 5 ) ,而仅在第 3周表现对Cytb5的诱导 (P <0 0 5 )。将玉米秸组和麦秸组的酶诱导作用与松木组进行比较 ,2～ 4周 ,玉米秸组CytP4 50 的值低于松木组的值 (P <0 0 5 ) ;2、3周麦秸组CytP4 50 的值低于松木组的

Abstract:Objective To observe the effect of lycopene(LP) on blood glucose and lipid peroxiditionin in diabetic rats. Methods The diabetic model was induced by alloxan in Wistar rats. The glucose oxidase method was used to determine blood glucose. The xanthines oxidase method and TBA were used to determine SOD and MDA in serum and pancreas respectively. {Results} Compared with model rats, blood glucose in the group of LP administration was decreased, SOD was increased and MDA was decreased significantly (%P%<0^05). Conclusion LP could effectively decrease blood glucose and increase antioxidative ability in the diabetic rats.;

Abstract:Twenty microsatellite DNA markers were selected from poultry genome in this study. The genomes of 100 Lingnan yellow chickens were analysed. the results from this study indicated that 13 of 20 microsatellite DNA markers were polymorphic.The number of alleles was 4-7, the average value of polymorphism information content was 0^6109 and the average value of heterozygosity was 0^6712.;

Abstract:Objective To study the distribution and morphology of 5-HT pisitive immunoreactive endocrine cells in the digestive tract of %Oryctolagus cunieulus% rabbits by the method of immunhistochemistry technique. Methods The method of the ABC immunhistochemistry technique was performed. Results There was negative reaction in esophagus and cardiac part, and positive reaction in the other parts. The cells' distribution of density was as !M". The shape of 5-HT positive immunoreactive endocrine cells was round and ellipse mainly, but shuttle with process in small intestine part and tapering in rectum. The cells were found in between epithelial cells and between gastric glands, but in the lamina propria in rectum. Conclusion The feeding habit and digestive character of %Oryctolagus cunieulus% rabbits affect the distribution of 5-HT positive immunoreactive endocrine cells in some extent, the morphology of 5-HT positive immunoreactive endocrine cells are conformable with the endocrine and exocrine functions.;

Abstract:目的研究松木、麦秸、玉米秸、玉米芯和国外商品垫料5种垫料物质的细胞毒性作用.方法将五种垫料物质的丙酮提取物按1、3、5、10、20和40mg/ml的终浓度梯度加入到H22-H2D8细胞中,培养72h后用考马斯亮蓝法测定细胞的总蛋白量,计算细胞的半数致死量(LD50);用MTT法测定H22-H2D8细胞在492nm的吸光度值(A492),计算细胞的相对增值率,用6级毒性分类法进行毒性分级.结果松木的LD50<5mg/ml,麦秸、玉米秸、国外商品垫料的LD50<20mg/ml,玉米芯的LD50>40mg/ml.麦秸和商品垫料的细胞毒性为2～3级,松木的细胞毒性2～5级,玉米秸的细胞毒性为1～3级,玉米芯的细胞毒性为1～2级.在垫料提取物浓度为5mg/ml、10mg/ml时,麦秸组的A492值显著低于玉米秸组和商品垫料组的A492值(P<0.05),而玉米秸组和商品垫料组相比无显著差异(P>0.05);在浓度为20mg/ml、40mg/ml时,王米秸组的A492值大于麦秸组和商品垫料组的A492值(P<0.05),麦秸组和商品垫料组相比差异无显著性(P>0.05).结论松木的细胞毒性最大,玉米芯的细胞毒性作用最小,玉米秸、国外商品垫料、麦秸三种物质均显示有细胞毒性作用,但它们的毒性显著低于松木,高于玉米芯.

Abstract:Objective The aim of this study was to isolate LaRV (lapine rotavirus) from dejection suspected of infection.Methods The isolate was determined by cellular morphology,biochemistry and molecular biology.Results The dejection suspected of infection contained virus of LaRV.Conclusion It was successful to isolate the strain of LaRV.;

Abstract:The flagellates,mainly pathogens Giardia muris and Hexamita muris and non|pathogens %Tricomonas spp.% and others in intestines of small laboratory animals,were reviewed for the reference of parasitology checkout of laboratory animals.;

Abstract:Objective To establish an model of experimental acute pancreatitis(AP) in rats by operational method.Methods Wistar rats were divided randomly into three groups.The end of bile duct was ligated in the test group;in the control group placket of duodenum was ligated one centimeter far and near to the bile duct;the normal group as a control:the normal Wistar rats.Test and control groups rats were phlebotomized at 10,24,48h respectively,for detection of the amylase concentration,calcium and blood sugar.At the same time,the pathological change and the histological change were examined.Results The level of amylase concentration of test and control groups increased significantly,and higher than the normal group as a control;the level of calcium reduced significantly,but blood sugar had no remarkable change.Conclusion Biochemical analysis and histological examination demonstrated that the AP model was successfully constructed.;

Abstract:Objective To investigate the toxicity of Caulis Aristolochiae Mansshuriensis(CAM).Methods 50 SD rats were divided into three groups:S and B groups which were given daily dose of 6g and 1g CAM in 2ml respectively,and control group,which fed with 2ml water.At 7 day after,urinary volume,urinary sugar,urinary protein,blood creatinine were measured,and B group was sacrificed for kidney pathological examination.At one and two month after,those of S and control groups were measured again.Of S group,10 rats were sacrificed for kidney check.At 3 month after,those did again,and all of them were sacrificed.Results 2 rats of B group died showing the kidney necrosis.The difference between S and control groups was of no significance (P>0.05).Conclusion High dose of CAM damaged the kidney,resulting in acute renal tubules necrosis.;

Abstract:The argyrophil cells in digestive tract of Pelophylax nigromaculata were studied with the silver nitrate method of Grimelius.The argyrophil cells could be observed throughout the digestive tract,distributing between epithelial cells of mucosa and epithelial cells of glands,with the highest distributive density in pyloricus and lowest in duodenum.The morphology of argyrophil cells differs in accordance with distributive parts.Argyrophil cells might have endocrine,exocrine,paracrine functions;and configuration of argyrophil cells might be related to the pressure.;

Abstract:Objective To establish an easy, economical, practical rat C6 glioma model.Method After fixation by self-made frame, C6 gliomaous cells were implanted into the specific plot of brain. The behavior and survival time of the rats were observed.Result After inoculation, diminished food intake, weight loss, intraocular hemorrhage, ocular proptosis, per se whirl, epileptic attack and hemiplegia ocurred in model rats. After 2-3weeks, pathological dissection showed oncogenesis in cranium.Conclusion The rat model of brain tumor we made is similar to the human's. The tumor is quick to form, and relatively unchangeable during the survival, so it can be used for studing the tumor's foundational mechanism, experimental cure and pharmacodynamics.

Abstract:Objective To establish an animal model of positive antisperm antibody (AsAb) in rats. Methods The animal model was set up by artificial immunity with iso-Wistar rat sperm antigen. The level of AsAb in serum was determined by ELISA. The morphological changes of the epididymis tissue in rats were determined by optical microscope with HE. The rats sperm was visualized by staining acrosome with the dye Coomassie brilliant blue. The changes of ATPase activity of the rats sperm were detected by chemical colorimetry. Results The AsAb of rats serum in the model accorded with the experimental demand. The percentages of acrosome reaction, Na -K ATPase activity, Ca 2-ATPase activity in AsAb positive rats sperm significantly decreased as compared with the control group (P<0.01; P<0.05; P<0.05).Conclusion The results demonstrated it was successful to establish an animal model of AsAb positive rats.

Abstract:目的 建立间接ELISA方法对兔轮状病毒抗体进行检测。方法 确定抗原和阴、阳性参考血清的最佳工作浓度；进行特异性、敏感性以及中间试验。结果 抗原的最佳工作浓度为1：12000倍；参考血清的最佳工作浓度为1：200倍；本次实验建立的ELISA方法具有很好的敏感性和特异性；中间实验表明，LaRV在兔群中具有很高的感染率。结论 本次试验成功地建立了用于兔轮状病毒抗体检测的ELISA方法。