人自然杀伤细胞抑制性受体P58.2基因克隆与鉴定

目的：P58．2基因克隆与鉴定。方法：采用RT—PCR技术，从正常人外周血单个核细胞中扩增自然杀伤细胞抑制性受体P58．2基因全长cDNA，经酶切后构建重组克隆载体，双酶切和测序鉴定。结果：RT—PCR获得了预期的扩增产物P58．2全长cDNA，成功构建了pSPORT1-P58．2重组克隆载体，酶切、酶谱分析与预期结果相符。DNA测序结果与GenBank登记的人P58．2cDNA全长碱基序列一致。结论：pSPORT1-P58．2重组克隆载体构建成功；P58．2基因序列与文献报道一致，为下一步构建表达载体和基因转染等工作打下良好的基础。     

A 1-year study of the epidemiology of hepatitis A virus in Tunisia

This 1-year (September 2000 to August 2001) prospective study investigated the presence of hepatitis A virus (HAV) in the population of Monastir, Tunisia (86 serum samples), in the influents and effluents of two wastewater treatment plants, and in shellfish harvested in the coastal areas of Monastir, Bizerte and Sfax (January 2001 to May 2001). The virus was detected by RT-PCR using primers targeted at the VP3-VP1 region. An epidemic of HAV infection was observed during the winter months, with a peak in January. The presence of the virus was relatively constant in the influents and effluents of the wastewater treatment plants, and the virus was found in shellfish from the Monastir area during the months of January and February. The genotype IA strain was recovered most frequently from human serum and wastewater samples. The observation that the peak of the epidemic was during the winter months suggests that transmission of HAV is related to climatic factors and, presumably, to shellfish consumption.     

卵巢癌细胞多种细胞因子基因表达的研究

本文应用RT-PCR方法,检测5例刚分离的晚期上皮性卵巢癌患者肿瘤细胞和3例卵巢痛患者腹水中肿瘤细胞IL-2、IL-2R、TNF-a,IL-6、TGF-p、IL-10等细胞因子基因的表达,用免疫学方法检测卵巢癌细胞上清液中IL-6活性。结果发现:卵巢癌肿瘤细胞表达IL-6mRNA和抑制性细胞因子TGF-p、IL-10。腹水中存在较多量lL-6可能来自肿瘤细胞。     

毒蕈碱样乙酰胆碱受体m1～m5亚基在大鼠前庭终器组织中的表达

目的：检测大鼠前庭终器中毒蕈碱样乙酰胆碱受体（mAChRs）m1～m5哑基的表达。方法：根据GeneBank公布的大鼠mAChR m1～m5亚基的mRNA序列设计特异性引物，再运用RTPCR方法检测大鼠前庭终器中编码mAChR m1～m5亚基的mRNA的表达，之后行PCR产物直接测序。以大鼠脑千组织提取RNA为阳性对照，以解剖液为阴性对照。结果：扩增出标志mAChR m1～m5亚基表达的PCR产物，并经测序证实与预计扩增的cDNA片段序列一致。结论：在大鼠前庭终器组织中有mAChR m1～m5亚基的表达，为乙酰胆碱作为大鼠外周前庭系统冲经递质提供了最直接的证据。     

Molecular epidemiology of norovirus variants detected in children under five years of age in Hyderabad,India

PurposeNoroviruses are common viral agents in acute diarrhea in all age groups worldwide. Norovirus has been classified into 10 genogroups, GI to GX with over 48 genotypes among them the GII.4 genotype has evolved over time with a clear pattern of periodic variant replacement. Immunity is strain or genotype specific with little or no protection conferred across genogroups. The present study was aimed to determine the epidemiology, prevalent genotypes of norovirus in children below five years of age in the Hyderabad region, India.MethodsThe stool samples and clinical data were collected from 458 children below 5 years of age comprising of cases with acute gastroenteritis (n ?= ?366) and a control group (n ?= ?92) admitted to the pediatric ward. All the samples were tested for Norovirus by ELISA and RT-PCR. Sequencing was done for predominant strains.Results10.3% (n ?= ?38) of cases and 3.2% (n ?= ?3) of the control group were found to be Norovirus positive. Predominant genotypes were GII-82.5% followed by GI-12.5%.ConclusionSequencing and Phylogenetic analyses of 20 GII.4 strains was done. All of the isolates are clustered away from published the GII.4 variants thus suggesting the appearance of a new variant.     

细胞因子Midkine融合蛋白的表达及其单克隆抗体的制备与应用

目的:克隆细胞因子midkine(MK)基因,表达其融合蛋白,制备抗MK单克隆抗体(mAb)并检测MK在肿瘤细胞中的表达。方法:利用MK基因的特异性引物,通过RT-PCR从人肾癌组织mRNA中扩增人MKcDNA分子。将其定向克隆于原核表达载体中,在大肠杆菌中表达MS2-MK融合蛋白。以纯化的融合蛋白免疫BALB/c小鼠,用传统的杂交瘤技术,进行细胞融合、筛选、克隆化并制备mAb腹水。用ELISA法分析其Ig亚类,用免疫细胞化学染色法检测MK在肿瘤细胞中的表达。结果:成功地克隆出MK基因并表达了MS2-MK融合蛋白。通过免疫和筛选,获得2株分泌小鼠抗人MKmAb的杂交瘤细胞,其分泌的mAb的Ig亚类分别为IgG1和IgG2a。免疫细胞化学染色显示,2株mAb与人胃癌细胞株MGC803和胃癌组织呈强阳性反应。结论:在原核细胞中获得MS2-MK融合蛋白的表达,并制备出抗MK的mAb,为研究MK的生物学活性提供了条件。     

Lumican基因在结直肠癌中表达的相关研究

目的探讨Lumican基因的表达与结直肠癌发生、发展之间的关系。方法应用RT- PCR方法检测49例结直肠癌组织及其癌旁正常黏膜上皮组织内Lumican mRNA的表达水平,并进行统计学分析。结果RT-PCR结果表明结直肠癌组织中Lumican mRNA均呈阳性表达,癌旁正常黏膜上皮组织内Lumican mRNA的表达显著低于癌组织(P<0.01)。Lumican mRNA的表达水平与患者的性别、年龄、组织病理分型、淋巴结转移及Dukes分期无关。结论结直肠癌组织中Lumican mRNA的表达增强提示Lumican mRNA在结直肠癌组织中的高表达对结直肠癌的发生发展可能起重要作用。