眼镜王蛇毒单克隆抗体的制备

目的:应用眼镜王蛇毒基因疫苗,免疫BALB/c小鼠,制备单克隆抗体(mAb).方法:应用眼镜王蛇Oh-3基因真核表达质粒pIRES-Oh3,佐以脂质体制成基因疫苗,免疫BALB/c小鼠,取其脾细胞与同源Sp2/0骨髓瘤细胞进行融合,经间接ELISA筛选,有限稀释法克隆,制备mAb.结果:获得了2株分泌抗眼镜王蛇毒mAb的杂交瘤细胞株(F5、 F11),细胞培养上清液的抗体效价分别为3.2×10-1、 6.4×10-1,腹水抗体效价分别为1.28×10-4、 2.56×10-4.结论:成功地制备了2株眼镜王蛇毒mAb.

乳腺增生病p53基因第6外显子突变检测

目的 :探讨p5 3基因在乳腺癌发生早期的作用及早期诊断乳腺癌的分子病理指标。方法 :用PCR -SSCP检测36例乳腺单纯性增生、 31例不典型增生、 30例乳腺癌中p5 3基因第 6外显子突变 ,用DNA直接测序技术确定突变的碱基及其所在的密码子。结果 :乳腺单纯性增生、不典型增生、乳腺癌中p5 3基因第 6外显子的突变率分别为 0、6 5 % (2 / 31)、 13 3% (4/ 30 )。 6个点突变均为碱基替换 ,其中 4个发生于第 192密码子 (CAG→TAG) ,2个发生于第2 13密码子 (CGA→TGA) ,两者均导致多肽链合成提前终止。结论 :乳腺癌不典型增生中存在p5 3基因第 6外显子突变 ,该突变可能在乳腺不典型增生发展到乳腺癌过程中起重要作用 ,可作为早期诊断乳腺癌的辅助指标。

period基因的Thr-Gly区段在果蝇和部分双翅目昆虫中的分子进化特征

以亲缘关系极近的近缘种类群、中等距离的远缘种类群为对象，分析生物钟基因period基因的Thr-Gly区段的分子进化特征，发现Thr-Gly区段在果蝇和部分双翅目昆虫中未曾经历性选择和其他定向的正选择。Thr-Gly区段在果蝇nasuta亚群中的分子进化速率为10.4*10~-9同义替换/位点/年。

Sequence variation and genetic diversity in the giant panda

About 336-444 bp mitochondrial D-loop region and tRNA gene were sequenced for 40 individuals of the giant panda which were collected from Mabian, Meigu, Yuexi, Baoxing, Pingwu, Qingchuan, Nanping and Baishuijiang, respectively. 9 haplotypes were found in 21 founders. The results showed that the giant panda has low genetic variations, and that there is no notable genetic isolation among geographical populations. The ancestor of the living giant panda population perhaps appeared in the late Pleistocene, and unfortunately, might have suffered bottle-neck attacks. Afterwards, its genetic diversity seemed to recover to same extent.

Structure analysis of CCR5 from human and primates

It is well known that the chemokine receptor CCR5 plays very important roles in HIV-1 virus infection. A three-dimensional molecular model of human CCR5 was generated by SYBYL, a distance geometry-based homologous modeling package, using the corresponding transmembrane domain of bacteriorhodopsin as the template. On the basis of human CCR5 model, we also built 18 3D molecular models of CCR5 in primates from Pongo pygmaeus, Pygathrix nemaeus, Macaca assameniss, Trachy-pithecus phayrei, T. francoisi, M. arotoides, Rhinopithecus roxellance, R, bieti, R. avunculus, Hylobates leucogenys, Pan troglodytes, Gorilla gorilla, Cercopithecus aethiops 1, C. aethiops 2, Papio hamadryas M. mulatta, M. fascicularis and M. nemestrina. Structural analyses and statistics results suggested that the main-chains of the primate CCR5 were similar to that of the human CCR5 and that the fit-RMS deviation values of these primate CCR5 were less than 0.1 Angstrom. Moreover, the structures of these CCR5 proteins, except those of the African green monkey 1 (C.aet1), do not have a remarkable difference. It is proved that the 14th residue is possibly very important in the inhibition infections by M-tropic HIV-1, and it is also demonstrated that the 13th residue of human CCR5 was changed from asparagine into aspartic acid in all these primates. It means that the primate CCR5 no longer depend on CD4 for efficient entry, but human CCR5 may have evolved subsequently due to the use of CD4 as a receptor, allowing the high-affinity chemokine receptor-binding site of HIV to be sequestered from host immune surveillance. (C) 2000 Elsevier Science B.V. All rights reserved.

贻贝凝集素抗-HIV活性研究

分离提取海洋无脊椎动物贻贝(Crenomytilus grayanus)凝集素,考察其抗HIV活性。采用半乳糖-Sepharose 6B亲和层析和Sephacryl S-200层析分离提取贻贝凝集素(Crenomytilus grayanus lectin,CGL),以光镜检查合胞体抑制试验,以ELSA测定HIVp24抗原表达水平。从海洋无脊椎动物贻贝中分离出的凝集素(CGL),为N-乙酰半乳糖胺/半乳糖(GalNAc/Gal)特异性的凝集素。CGL在27.88mg.L-1浓度时,对HIV诱导细胞病变的抑制达50%;在45.70mg.L-1时,对HIV-1复制的抑制达50%;同时在35.12mg.L-1浓度时,对HIV感染细胞融合的阻断达50%。

New rotenoids from roots of Mirabilis jalapa

Four new rotenoids named mirabijalone A-D-1) (1-4), together with 9-O-methyl-4-hydroxyboeravinone B (5), boeravinone C (6) and F (7), and 1,2,3,4-tetrahydro-1-methylisoquinoline-7,8-diol (8), were isolated from the roots of Mirabilis jalapa. The structures of these compounds were determined on the basis of their HR-EI-MS, IR, UV, H-1- and C-13-NMR (DEPT). and 2D NMR (HMQC, HMBC, NOESY) data. Among them, 1,2,3,4-tetrahydro-1-methylisoquinoline-7,8-diol (8) showed a 48% inhibition against HIV-1 reverse transcriptase at 210 mug/ml.

芒果柑桔小实蝇[Dacus (Bactrocera) dorsalis (Hendel)]生物学及其综合防治研究

芒果柑桔小实蝇是云南芒果主要害虫之一, 幼虫取食芒果果肉, 对3年芒果危害率为32.4％, 对象牙芒果为34.7％。该虫在云南景谷地区1年发生5代, 世代重叠, 全年均有活动, 以老熟幼虫、蛹入表土中或成虫栖于杂草丛中越冬。采取综合防治措施后, 3年芒果危害率降为2.3％, 象牙芒果降为1.8％。

懒猴能量需求的研究

三组成年懒猴(Nycticebus coucang bengalensis)日均摄入量能值分别为210.19kJ/kG, 177.222kJ/kG和161.608kJ/kG; 日均消化能分别为191.89kJ/kG, 155.131kJ/kG 和146.783kJ/kG。据统计学方差及t测验分析表明:I—Ⅱ组之间和I—Ⅲ之间的日 均摄食量能值及日均消化能均有显著性差异。实验结果表 明: I组所饲喂的配 合饲料的组成优于Ⅱ、Ⅲ组并可做为懒猴 的长期用配方。表4参8

弥散张量成像在精神分裂症的研究

　弥散张量成像(diffusion tensor imaging , DTI) 是目前在活体直视脑白质主要纤维束三维结构的唯一方法〔1〕。国内外已将其应用于神经精神疾病的研究。本文就DTI 在精神分裂症的应用作一综述。

树鼩细胞周期蛋白T1 cDNA的克隆和序列分析

树鼩细胞不能感染 HIV-1, 但支持 HIV-1进入靶细胞后的转录, 可能是因为树鼩细胞周期蛋白T1(tsCycT1)能结合HIV-1的 Tat 蛋白. 通过设计引物, 用 RT-PCR 技术, 获得全长为2175bpts CycT1 基因的cD NA. 其核苷酸序列与人的 CycT1(hCycT1) 基因的 cDNA 有92.6%的相似性; 由此推导出的氨基酸序列有94.1%相似性. 其中, hCycT1 和 tsCycT1 氨基端的1～272个氨基酸的相似性高达98.8%, 氨基酸第261位点为半胱氨酸. 这些结果提示, tsCycT1 会和 HIV-1 的 Tat 结合, 形成高亲和的、锌依赖的复合物, 支持 HIV-1转录。