小鼠轮状病毒EW株VP7基因的克隆及其在重组腺病毒中的表达

目的：构建表达小鼠轮状病毒(EDIM)EW株VP7基因的重组腺病毒，以在小鼠模型上研究轮状病毒的免疫保护机制。方法：用RT-PCR方法扩增EDIM VP7全长cDNA并进行基因序列分析，将所得的VP7基因片段插入腺病毒穿梭质粒pShuttle-CMV，获得重组质粒pShuttleCMV-EVP7，将该质粒与腺病毒骨架质粒pAdEasy-1共转化大肠杆菌BJ5183获得重组腺病毒质粒pAdCMV-EVP7，将该质粒线性化后转染293细胞包装重组腺病毒rvAdEVP7。以rvAdEVP7感染293细胞，并分别应用电子显微镜、PCR、RT-PCR和Western blot等方法观察rvAdEVP7的形态、VP7基因整合、遗传稳定性、VP7基因在细胞内转录及表达等有关生物学特性。结果：获得了小鼠轮状病毒的全长cDNA，重组腺病毒rvAdEVP7具有典型的腺病毒形态，PCR、RT-PCR和Western blot等分子生物学方法分析显示：rvAdEVP7中确有特异性的VP7基因稳定整合；有较好的遗传稳定性；感染293细胞后能有效转录；可表达轮状病毒主要结构蛋白VP7。结论：成功构建含VP7基因的重组腺病毒rvAdEVP7，为进一步研究轮状病毒的免疫保护机制打下了基础。     

脑动静脉畸形血管重构的分子生物学机制

脑动静脉畸形存在活跃的血管重构,本文就其重构的分子生物学机制作一综述。     

抗凝治疗肺心病血栓前状态的临床研究

目的探讨肺心病患者的血栓前状态，观察并分析低分子肝素抗凝干预的疗效。方法将54倒肺心病患者分为低分子肝素治疗组和常规治疗组各27例。分别检测治疗前后血浆血管性假血友病因子(VWF)、血浆凝血酶片段1 2(F1 2)、纤堆蛋白原(Fg)、血小板颗粒膜蛋白(GMP-140)和D-二聚体(DD)。结果肺心病患者VWF、F1 2、Fg、GMP-140、DD血浆浓度显著高于对照组；肺心病治疗组经LMWH治疗后上述各凝血分子标记物及PaCO2明显降低。而PaO2显著增高。常规组虽血气指标略有改善。但凝血分子标记物未见改善。结论尽早检测肺心病血栓前状态，及时给予低分子肝素抗凝干预，有望从根本上改善肺心病惠性加重期患者的不良预后。     

Loss of constitutional heterozygosity in human astrocytomas

Summary Inactivation of tumour suppressor genes or anti-oncogenes as well as activation of dominant acting oncogenes seem to be important mechanisms in the pathogenesis of gliomas. We compared constitutional and tumoural genotypes at different restriction fragment length polymorphism loci (RFLP) on chromosomes 10 and 17 in 15 unrelated individualsLoss of heterozygosity (LOH) pointing to chromosomal loss or deletions was detected for at least one chromosme 17 marker in 11 gliomas (astrocytomas grades I–III and glioblastoma multiforme), whereas LOH for chromosome 10 loci was only detected in 3 out of 9 cases of glioblastoma multiforme and was not detected in low grade gliomas. Since LOH for chromosome 10 loci seems to be restricted only to glioblastoma multiforme, it is possible that recessive mutations on chromosome 10 are engaged in tumour progression from astrocytomas to glioblastoma multiforme. As LOH of chromosome 17 markers occurs in astrocytomas as in glioblastoma multiforme, chromosome 17 loci probably are involved in early tumour development.     

展望分子谱系地理学在道地药材研究中的应用

作者在介绍分子谱系地理学的概念、研究方法和研究现状的基础上,探讨了分子谱系地理学推断的3种植物遗传分化模式--异域片断化、受距离影响的有限基因流和分布区快速扩展与道地性形成的相关性;阐述了基于分子谱系地理学研究的道地药材分子鉴定对以往分子鉴定局限的突破;阐明了分子谱系地理学在道地药材产地变迁历史研究中的应用;论述了分子谱系地理学在解决道地药材栽培中种质退化问题等方面的作用.这些方面的应用展现了分子谱系地理学在道地药材研究中的应用前景,为道地药材的研究提供了新的理论和方法.     

A new nematode, <Emphasis Type="Italic">Pongobius hugoti</Emphasis> gen. et sp. n. from the orangutan <Emphasis Type="Italic">Pongo abelii</Emphasis> (Primates: Hominidae)

Summary Light and scanning electron microscopical examination and molecular analysis of pinworm samples collected from the Sumatran orangutan (Pongo abelii) from Sumatra (Indonesia) revealed the presence of a new taxon, Pongobius hugoti gen. et sp. n. (Oxyuridae: Enterobiinae). The monotypic genus Pongobius is characterised by the presence of a triangular mouth, three rounded teeth and three chisel-like inner teeth in the buccal cavity, an oesophagus with a long pharyngeal part, corpus posteriorly slightly enlarged and continuing as a pyriform bulb, isthmus absent. These features differentiate P. hugoti from all members of the subfamily Enterobiinae. The male has a long caudal appendix (> 50% of total tail length). Molecular analysis of the new taxon is presented for more precise identification.     

结直肠癌淋巴结转移与部分肿瘤分子标志物表达的相关性研究

目的 探讨部分肿瘤相关分子标志物免疫组织化学的表达与结直肠癌淋巴结转移的相关性.方法 应用免疫组织化学技术检测65例结直肠癌手术标本Ki-67、p53的表达情况,对照手术所见和手术标本的病理检查结果 ,研究这些肿瘤相关分子标志物与肿瘤的生物学特性如浸润和淋巴结转移等的关系.结果 65例结直肠癌手术标本Ki-67、p53免疫组织化学的表达与肿瘤肠壁浸润深度无明显相关性(P＞0.05).Ki-67的表达及Ki-67标记指数的表达与淋巴结转移及Dukes分期有明显的相关性(P＜0.01);p53标记指数的表达与淋巴结转移有相关性(P＜0.05),与Dukes分期有明显的相关性(P＜0.01).结论 作为反应细胞增殖活性的肿瘤相关分子标志物Ki-67,其免疫组织化学的表达程度可间接反映结直肠癌淋巴结转移状况,可能成为反映淋巴结转移的一个标志物.     

Construction of Eukaryotic Expression Plasmid of Human PRX3 and Its Expression in HEK-293FT Cells

To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could he expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.     

sFGFR1基因的克隆与在RTS系统中的高效表达

目的：克隆sFGFRl(soluble fibroblast growth factors receptor-1)基因，并在RTS(rapid translation system)系统中高效表达相应蛋白．方法：培养Swiss rat 3T3 fibroblast细胞株，提取总RNA，用RT—PCR方法获取鼠sFGFR1 cDNA片段，酶切后克隆到pIVEX2．3d载体并进行序列分析；采用Roche RTS ProteinMaster500系统，高效表达sFGFR1蛋白并用Western Blot鉴定表达的蛋白．结果：克隆了sFGFR1基因，测序证实序列正确；Western Blot证实sFGFR1蛋白在RTS系统中高效表达．结论：克隆了sFGFR1基因并在RTS系统获得高效表达．