Abstract:目的　研究单纯疱疹病毒胸苷激酶 (HSV1 TK)基因转染并联合抗病毒药更昔洛韦 (Ganciclovir ,GCV)对人胶质母细胞瘤细胞的杀伤效应。方法　采用基因工程技术构建带HSV1-TK基因的逆转录病毒重组体pLX SN TK ,采用脂质体介导入PA317包装细胞 ,建立重组逆转录病毒载体分泌细胞株PA317 TK ;用该细胞上清液转导人胶质母细胞瘤细胞 ,用不同浓度GCV作用人胶质母细胞瘤细胞SW0 38 C2 TK和野生型SW0 38 C2 ,采用噻唑蓝(MTT)比色法检测 72h后细胞存活率 ,并求出半杀伤浓度。结果　重组逆转录病毒载体pLXSN TK能有效地将TK基因导入SW0 38-C2细胞内 ,并使其获得对GCV的敏感性 ;体外细胞毒实验 :在 2 0 μmol LGCV存在的情况下 ,野生型细胞无明显改变 ,而实验组细胞明显死亡 ,半杀伤浓度IC50 为 0 8μmol L ,与SW0 38 C2相比 ,对GCV的敏感性提高了 6 0 0倍 ,旁杀效果也较明显。结论　SW0 38 C2转染TK基因后 ,能有效地被GCV杀灭。显示了其潜在的临床应用价值

Abstract:目的　原核胚是转基因等生物技术所必需的主要材料 ,其冷冻保存使操作不受时间和空间的限制。另外 ,冷冻保存还可以避免体外培养过程中的细胞发育阻断期。方法　在室温 (2 0℃或 2 5℃ )条件下 ,以乙二醇、DMSO为主体抗冻保护剂配制成的玻璃化溶液 (EFS、EDT、EDFS) ,不借助冷冻仪 ,对小鼠原核胚进行一步法和二步法玻璃化冷冻保存。结果　 2 0℃室温条件下 ,用EDFS4 0平衡 1min一步法冷冻解冻后的原核胚 ,经培养后囊胚发育率最高仅为 4 7% ,与新鲜原核体外培养的对照组 (75 % )的差异极显著 (P <0 0 1) ;当原核在 10 %E +10 %D溶液中预处理 5min ,移入EDFS30中平衡 30s二步法冷冻保存 ,解冻后的囊胚发育率达 6 8%。而室温升至 2 5℃ ,二步法冷冻保存后原核胚的囊胚发育率高达 77% ,与对照组差异无显著性 (P >0 0 5 )。用最佳冷冻组的原核胚或解冻后培养到囊胚移植给受体母鼠均获得产仔。结论　本研究对小鼠原核胚实施玻璃化冷冻保存 ,经体外培养和移植结果与对照组无显著性差异 ,证明了本方法的可行性

Abstract:目的　研究实验小鼠H - 2抗原单克隆抗体在初建时和经冷冻保存后的效价变化规律 ,为长期保存小鼠H - 2抗原单抗及规模性推广这一遗传监测方法奠定技术基础。方法　制备 6种抗小鼠H 2抗原的单克隆抗体 ,将具备一定抗体效价 (16 0倍以上 )的细胞株保存在 - 196℃的液氮中 ,数月后将复苏后的部分细胞株培养 4 8h后 ,取其上清液用微量细胞毒试验法测定抗体的效价 ,并与其原有效价进行比较。结果　单抗初建时效价达到 16 0倍以上的细胞株占总株数的 37%～ 5 3% ,H 2K抗体复苏后效价持平的细胞株占 13% ,效价下降的为 74 % ,效价上升的是 13% ;H 2D抗体效价持平的细胞株占 5 3% ,下降的为 31% ,上升的是 16 %。复苏后的抗体效价大部分与原效价持平或略有下降 ,个别的会有所提高。结论　单克隆抗体的效价略低于同种单价特异性抗血清 ;能否使冻存后的抗体保持原有的效价是抗体能否大量生产并推广的关键 ;要长期保存和生产H 2抗原单抗 ,应加强克隆和检测工作 ,建立稳定的制备、保存和检测系统 ,才能为实验动物的遗传监测提供技术服务

Abstract:目的　观察博来霉素 (Bleomycin ,BLM)致大鼠肺纤维化模型的形态学及胶原含量的变化 ,探讨其发生机制。方法　健康SD大鼠 ,BLM 5mg (kg·bw)复制大鼠肺纤维化模型 ,于 1、3、7、14、2 8d ,观察肺纤维化形成的病理变化和肺匀浆羟脯氨酸 (HYP)的含量变化。结果　光镜所见 :给予BLM后 1- 7d肺部病变以肺泡炎为主 ,14d以后则进入慢性纤维化阶段。电镜所见 :模型组 3- 7dⅠ型细胞受损 ;Ⅱ型细胞增生 ,其内板层小体明显增多 ,线粒体嵴消失甚至出现空泡样变 ;14 - 2 8dⅡ型细胞数目减少 ,纤维组织增生。模型组肺组织匀浆HYP含量于 7d开始明显增加 ,2 8d达高峰。结论　博来霉素诱发大鼠肺纤维化早期以肺泡炎为主 ,7d开始出现肺纤维化 ,2 8d肺纤维化为主要病变

Abstract:Objective To analyze the differences between the two kinds of hepatic fibrotic animal models in time effect and pathologic features in the course of development of liver fibrosis. Methods\ CCl 4 and PS ten week methods were used to make rat models of liver fibrosis. The development of pathologic features of liver fibrosis was observed at 6, 10, 14 and 20 weeks of model making respectively, and these presentations were analyzed by Masson stain and computerized image analyzing system. Results\ In CCl 4 method group, fibrotic pseudolobuli developed at 6 weeks during model making, became typical at 10 weeks, and got tranquilization thereafter, with ballooning degeneration of liver cells outstanding. On the contrary in PS method group, fibrosis occurred late in 10 weeks during model making, and developed from 10 weeks through 14 weeks though the model making had stopped; meanwhile ballooning degeneration was not presented. Conclusion\ There was indeed something different in the development of liver fibrosis in time effect relation and pathologic features between the two groups of model making. This indicated different mechanisms in the pathogenesis of liver fibrosis. The two methods of model making should be combined in the coming animal study of liver fibrosis; the prophylactic and treatment studies by anti fibrotic drugs should last no less than 14 weeks, and should begin 4 weeks ahead of the beginning of and the end of model making.

Abstract:Objective To study expression and location of a novel gene that might be related to cell differentiation in normal tissues,cancer tissues and cell lines.Methods\ Northern blotting method was used to detect the expression of the gene in 10 embryo tissues,6 cancer cell lines,4 cancer tissues.The immunohistochimical method was used to detect the gene location.Results\ The gene showed high level expression in embryo tissue,cancer tissue and cell line of human,the expression in normal tissue and paratumor tissue decrease obviously.The gene is located on the membrane of K 562 cell.Conclusion\ The gene may play an important role in cell differentiation and carcinogenesis.

Abstract:目的　探讨一氧化氮供体—硝普纳 (SodiumNitroprusside ,SNP)对昆明小鼠卵母细胞体外自发成熟的影响。方法　利用体外培养方法 ,在培养的不同时间观察卵母细胞的成熟情况 ,研究SNP对卵母细胞自发成熟的动力学影响。结果　 (1) 1mmol LSNP能够明显延迟卵丘卵母细胞复合体 (CEOs)的自发成熟 ,与对照组相比 ,CEOs的生发泡破裂 (GVBD)和第一极体 (PB1)释放分别延迟了 8h和 10h。但在培养结束 (2 4h)时 ,CEOs处理组的PB1释放率与对照组相比有明显的降低 ,而处理组的生发泡破裂 (GVBD)百分率和对照组相比没有明显变化 ;1mmol LSNP能够明显延迟裸卵母细胞 (DOs)的自发成熟 ,与对照组相比 ,DOs处理组的GVBD和PB1释放分别延迟了 6h和 12h。且在培养 2 4h后 ,DOs处理组的GVBD和PB1释放率与对照组相比有显著的降低。 (2 ) 1mmol LSNP能够明显促进CEOs中卵丘细胞的离散 ,造成CEOs互相粘连在一起。 (3) 1mmol LSNP能够明显影响体外培养的DOs的形态 ,但对CEOs的卵母细胞却没有影响。而 1μmol LSNP对CEOs和DOs的自发成熟都没有影响。 结论高浓度的SNP对小鼠卵母细胞体外自发成熟有抑制作用

Abstract:Objective To explore the application of multiplex PCR method in the detection of microsatellite in mice. Methods\ Four pairs of specific primers were selected to perform multiplex DNA amplification in ten inbred stains of mice. Results\ Duplex DNA amplification got two expected bands under the same condition of the PCR reaction, but triplex DNA amplification did not get three expected bands.Conclusion\ The duplex DNA amplification is a fast and economical method to analyse genotype of inbred mice.

Abstract:目的　以不同性腺发育阶段的雌性鲤鱼为材料 ,采用脑垂体碎片离体灌流孵育的方法 ,研究 17β 雌二醇对由神经内分泌因子—促性腺激素释放激素和促甲状腺素释放激素诱导的生长激素分泌的调节作用的季节性变化特点。方法与结果　用 10 - 8mol L和 10 - 1 0 mol L 17β 雌二醇对鲤鱼脑垂体碎片进行 10～ 12h的过夜孵育后 ,性腺处于退化期和发育期的的鲤鱼脑垂体对浓度依次为 10 - 9,10 - 8和 10 - 7mol L的促性腺激素释放激素类似物的脉冲刺激的反应性显著大于对照组 ,但在成熟期 17β 雌二醇作用组的反应性与对照组无显著性差异 ;经过同样浓度的 17β 雌二醇预孵育后 ,鲤鱼脑垂体碎片对浓度依次为 10 - 1 0 ,3× 10 - 8和 10 - 7mol L的促甲状腺素释放激素脉冲刺激的反应性也表现出季节性差异 ,反应性的增强作用表现为 :成熟期 >发育期 >退化期。结论　性类固醇激素具有调节脑垂体对刺激生长激素分泌的神经内分泌因子的反应性的作用 ,这种调节作用与鱼体的性腺发育状态有关 ,具有季节性变化的特点

Abstract:目的　建立饮食诱导新西兰兔糖尿病模型 ,探讨脂肪毒性和葡萄糖毒性在 2型糖尿病发病中的意义。方法　将雄性新西兰兔 30只按血糖血脂浓度随机分为 2组 ,15只饲以基础饲料作为正常对照组 (C组 ) ;15只饲以高糖高脂饲料作为实验组 (DD组 ) ,共观察 32周 ,每 4周从禁食过夜的兔耳静脉抽取血样 ,测定血清中血糖、胰岛素、甘油三酯。于 32周时 ,全部处死动物 ,取动物胰腺。取部分胰腺用 10 %中性甲醛液固定 ,常规石蜡包埋切片 ,H、E染色 ,光学显微镜下观察 ;另一部分胰腺用 2 5 %戊二醛固定 ,常规电镜样品制备 ,透射电镜下观察并照相。结果　实验组 4周后即出现高血糖、高甘油三酯 ,并随着喂养时间延长 ,而有所升高 ,基础饲料组血糖、甘油三酯未见明显升高 ,两组比较差异具有显著性意义 (P <0 0 1)。以不同喂养时间作方差分析 ,整体上差异具有显著性意义。实验组血清胰岛素水平整体上差异没有显著性意义 (P >0 0 5 )。实验组胰腺组织表现为胰岛萎缩 ,胰岛边缘皱缩 ,胰岛数目减少 ,胰岛内细胞数量亦减少 ,多数细胞呈梭形 ,细胞核呈杆状 ,胰岛周围少量淋巴细胞浸润。透射电镜下观察到实验组胰岛 β细胞体积略小 ,核较小 ,部分分泌颗粒呈空泡状 ,细胞内可见局部结构较模糊 ,粗面内质网少 ,未见明显淀粉样变

Abstract:Objective To observe the effect of LINGGUANG injection(compound anisodine)on the mucosa of gastrointestinal tract after hemorrhagic shock in rats Methods 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 hemorrhagic shock at 4 kPa blood pressure with or without LINGGUANG injection treatment Half of the rats were sacrificed at 6 hour after resuscitation,the rest were killed at 12 hour Some organs were qualitatively cultured for translocating bacteria Stomachs,jejunums and ileums were examined under light microscope, and jejunum mucosas were observed under transmission and scan electron microscope Results and Conclusion Bacteria translocating to mesenteric lymph nodes,livers and spleens was less when treated with compound anisodine compared with control shock rats,the damages of gastrointestinal tract also became slighter in treated rats The mechanisms of compound anisodine treatment may be related with microcirculation improving and free radical scavenging

Abstract:Objective To observe the different cell surface characteristics of the breast carcinoma in closed breeding Microtus fortis and Kumming mouse.Method Tissues were observed under the scanning electron microscope. Results There were a number of folds and very small irregular popular processes on the surface of normal mammary gland cells of Microtus fortis ;while the surface of breast carcinoma cell in Microtus fortis was densely covered with a large number of long and irregular microvilli. In Kunming mouse, the cell surface of its breast carcinoma was also covered with microvilli, whose morphologic characteristics were very distinct and the end of them was round in shape. Conclusion There are distinguishing characteristics of microvilli on the surface of the breast carcinoma cells between Microfus fortis and Kunming mouse.

Abstract:目的　研究不同鼠龄组组间及组内肝腺泡Ⅰ、Ⅲ带肝细胞rDNA活化之形态计量的差异。方法　以甲酸明胶硝酸银混合制成Ag NORs染色液 ,活化的rDNA被染成深棕色 ,Ag NORs颗粒由Mais 2 0 0 0图像分析仪对各鼠龄组 (3、13、2 5、33月 )肝腺泡Ⅰ、Ⅲ带肝细胞核之Ag NORs分别进行了Ag NORs数、平均灰度与核面积比的形态计量测定。结果　Ⅲ带肝细胞核Ag NORs平均数 ,以 33月龄组分别少于 3、13、2 5月龄组 (P <0 0 5 )。Ag NORs平均灰度以 33月龄组分别小于 3、13、2 5月龄组 (P <0 0 5 )。结论　 33月龄肝腺泡Ⅲ带肝细胞核rDNA活性分别较 3、13、2 5月龄低

Abstract:Objective To understand the injury mechanism of Vero cells(African green monkey kidney cells) exposing to chloroform(CCl 3)for 24hours.Methods\ The content of reactive oxygen species(ROS) and cytoplasmic Ca 2+ concentration in Vero cells were measured with microfluorometry. Meanwhile, the injury of Vero cells was detected with lactate dehydrogenase(LDH) activity in the supernatant of the medium for Vero cells. Results\ There were no any differences for the above indices between CCl 3 group of 4 0mM concentration and control group statistically( P >0 05). Moreover, there were no any differences in cytoplasmic Ca 2+ concentration of Vero cells between all groups exposing to CCl3 and control group( P >0.05). In the other hand, the contents of ROS in the Vero cells exposing to CCl 3 at 8 0mM and 12 0mM concentration were higher than those in the control group( P <0 01)respectively, and the activity of LDH in supernatant of the medium for Vero cells exposing to CCl 3 at 8 0mM and 12 0mM concentration was higher than that of latter( P <0.05, P <0 01)respectively. Conclusion\ The possible injury mechanisms of Vero cells exposing to CCl 3 of high concentration are partly mediated by increasing the content of ROS in Vero cells.

Abstract:Objective To establish the inbred strain of the coagulation factor IX knockout mice. Methods\ The mice were bred by brother sister first mating with reproductive performance recorded.The homogeneous mice were selected after F8 Results\ Factor IX knockout mice were developed in the 12th generation.The age of mating was 70-90 day.The average birth litter size was over 5,average weaning litter size was over 3 Conclusion\ The coagulation factor IX knockout mice can be generated steadily.

Abstract:Objective\ To explore the effect on left ventricle haemodynamics of Wistar rat caused by complex factors including slimming, forced running and large dosage of propanolol.Methods Every animal in experiment group was forced to run with slimming. From 17th day ig propanolol was given and lasted for 4 days. At 21st day haemodynamics was measured in 6 rats. Then the rest rats were divided into supplement Qi group and nourishment Yin group. Supplement Qi group was given ig \%Buxinqi\% oral liquid for 12 days. Nourishment Yin group ig \%Zixinyin\% oral liquid for 12 days. In the end, haemodynamics measured.Results The indexes of experiment group such as intraventricular pressure max ,+dp/dtmax mean ,Vpm mean were less than those of control group significantly. The indexes of experiment group such as intraventricular pressure min ,-dp/dtmax mean were higher than those of control group significantly.Conclusion The complex factors including slimming, forced running and large dosage of propanolol could restrain the heart function of Wistar rat.