Detection of thiopurine s-methyltransferase mutations by template directed determinator in incorporation with fluorescence polarization （TDI-FP）

Objective: To develop a new method for the detection of TPMT gene mutations and determine the frequencies of four TPMT alleles, TPMT^*1,^*3A,^*3B and ^*3C in a healthy Chinese population. Methods:A TDI-FP assay system was set up in out lab. To evaluate this system, 220 healthy individuals were analyzed for the polymorphic sites at positions 460 (G→A)and 719 (A→G)of the TPMP gene using our new TDI FP method. Results:Three TPMP *3C(G^460→G^719) heterozygotes were identified, TPMP ^*3A and TPMP ^*3B were not found. All mutations were confirmed by conventional DNA sequencing analysis. Conclusion:TDI-FP method has proven to be very efficient as a rapid and accurate approach for TPMP genotyping. TPMP ^*3C was the only polymorphism identified in this clinical samples we have registered.     

苏州地区汉族人血小板糖蛋白Ⅲa基因TaqI多态性与脑梗死的相关性研究

目的　研究苏州地区汉族人血小板糖蛋白Ⅲa基因TaqⅠ多态性与脑梗死发生的相关性。 方法　应用PCR技术结合TaqⅠ酶切分析、聚丙烯酰胺凝胶电泳 ,研究了苏州地区汉族正常人与脑梗死患者血小板糖蛋白Ⅲa基因TaqⅠ多态性特点。 结果　正常对照组TaqⅠ等位基因的频率分别为C 5 3.8%、A 4 6 .2 %。脑梗死组TaqⅠ等位基因频率分别为C 5 0 .6 %、A 4 9.4 %。脑梗死组和对照组之间无显著性差异 (P >0 .0 5 )。结论　苏州地区汉族人血小板糖蛋白Ⅲa基因TaqⅠ多态性与脑梗死发生无关。     

白细胞介素-6基因启动子多态性与子宫内膜异位症的相关性分析

目的 研究白细胞介素-6(IL-6)基因启动子-174G/C、-572C/G及-597G/A的多态性在子宫内膜异位症(EM)患者及非EM育龄女性(对照组)中的分布.方法 采用病例对照研究的方法 ,选取98例EM患者,其中轻、中、重型(即Ⅱ、Ⅲ、Ⅳ期)分别为22、35、41例,以99名非EM育龄女性外周血白细胞为对照,应用聚合酶链反应-限制性片段长度多态性技术检测IL-6基因启动子-174、-572及-597的多态性,比较两组IL-6启动子基因型和等位基因的分布频率,对两组间基因多态性存在明显差异的基因位点进行组间比较.结果 IL-6基因启动子-174位点的GG、CG和CC基因型在EM组中分别为100%、0%、0%,在对照组中分别为98.99%、1.01%、0%,两组的分布频率差异无显著性;-572位点的CC、CG和GG基因型在EM组中分别为55.10%、42.86%、2.04%,在对照组中分别为70.71%、28.28%、1.01%,两组的基因分布频率差异有显著性;所有实验对象-597位点的基因型均为GG型,两组均无GA及AA基因型存在;-572位点的CC、CG和GG基因型在轻型EM组中分别为59.09%、40.90%、0%,在中型EM组中分别为54.29%、42.86%、2.85%,在重型EM组中分别为53.66%、43.90%、2.44%,轻、中、重组间基因型及等位基因分布频率无明显差异.结论 IL-6基因启动子的-174位点变异率极低,-597位点末发现变异,以上两位点基因变异与EM的发病无明显相关性;而启动子-572位点的多态性分布与EM的发病具有一定程度的相关性,但与疾病的严重程度无关.     

Genetic susceptibility to type 2 diabetic nephropathy in human and animal models

SUMMARY:   Diabetic nephropathy is the most common cause of end-stage renal disease (ESRD) in Japan, Western Europe, and the United States. Mega studies such as Diabetes Control and Complication Trial (DCCT), Epidemiology of Diabetes Interventions and Complications (EDIC), and the United Kingdom Prospective Diabetes Study (UKPDS) clarified that poor glycemic and blood pressure control are undoubtedly involved in the development of nephropathy. However, these factors are not sufficient to predict which diabetic patients will develop renal disease, because not all patients with poor glycemic and blood pressure control develop renal disease. Since ethnic variations and familial clustering of diabetic nephropathy have been observed, genetic factors might contribute to susceptibility to this disease. Several methods such as (genome wide) association studies, sib-pair analysis, and quantitative trait loci (QTLs) analysis are available to examine polygenic diseases. However, no mutations that could explain the majority of nephropathy cases have been identified so far. The development of most diabetic nephropathy might be explained by the polygenic effect (i.e. many minor gene-gene interactions might be very important in the development of nephropathy). Identification of candidate genes of nephropathy enables targeting of therapy in patients at risk and development of novel therapeutic agents.     

Common human Toll-like receptor 9 polymorphisms and haplotypes: association with atopy and functional relevance

BACKGROUND: Toll-like receptor 9 (TLR9) is a pattern-recognition receptor that detects unmethylated CpG motifs prevalent in bacterial and viral DNA. TLR9 stimulation is a key event after bacterial infection, triggering innate immunity and T-helper type 1 skewed adaptive immunity. Synthetic CpG-oligodeoxynucleotides (CpG-ODNs) represent a promising and novel class of immune adjuvants for allergy treatment, vaccination, and cancer therapy. However, common functional TLR9 gene variants could interfere with the clinical utilization of CpG-ODN in immunotherapy. Recently, a possible association of TLR9 polymorphism C-1237T with asthma has been reported. OBJECTIVE: The aim of the present study was to investigate whether TLR9 polymorphisms or haplotypes have functional relevance and are associated with atopy. METHODS: We genotyped five common TLR9 single-nucleotide polymorphisms (SNPs) in promoter, exon, and intron regions of the gene in 527 healthy blood donors, and estimated four common haplotypes. The total IgE and specific IgE levels against the most common aeroallergens were measured (n=303). IFN-alpha production by plasmacytoid dendritic cells (pDCs) was analysed after stimulation with TLR9 ligand CpG-ODN (n=220). RESULTS: No significant influence of common TLR9 polymorphisms and haplotypes on the total and specific IgE levels was found. Functional analysis of CpG-ODN-induced IFN-alpha did not indicate a significant role for common TLR9 gene polymorphisms in TLR9 function. CONCLUSION: We conclude that common genetic differences in the TLR9 gene exert no major influence on allergy susceptibility, and are unlikely to have on impact on clinical application of CpG-ODNs.     

代谢酶基因多态性与胰腺癌易感性研究进展

代谢酶基因多态性与肿瘤的关系是近来国内外研究的热点。代谢酶基因多态性使酶的活性发生改变，是胰腺癌遗传易感性的重要机制。现综述了与胰腺癌癌变过程中有关的代谢酶基因多态性与其易感性的关系。     

载脂蛋白A1、B基因多态性对非创伤性股骨头坏死发生的影响

目的:探讨载脂蛋白A1、B基因多态性对非刨伤性股骨头坏死(avascular necrosis of the femoral head,ANFH)发生的影响.方法:应用聚合酶链反应对中国北方汉族143例ANFH患者和92例正常人分别扩增含Apo AI基因启动子-75 bp和第一内舍子 83 bp及Apo B基因Eco RI、XbaI和3-VNTR的DNA片段,限制性内切酶酶切扩增产物,琼脂糖凝胶电泳分离基因多态性.结果:Apo A1基因启动子-75 bp处,ANFH患者中A/A基因型频率明显高于正常组(P＜0.01),而G/A基因型频率明显低于正常组(P＜0.01).Apo AI内舍子 83 bp位点,Apo B基因Eco RI、Xba I位点和3-VNTR区域ANFH患者组和正常组基因型及等位基因频率分布无统计学差异.结论:Apo A1基因启动子区域-75bp位点A/A型可能是非创伤性股骨头坏死易感基因之一,但未能发现Apo A1第一内舍子 83 bp位点及Apo B基因Eco RI、XbaI和3-VNTR位点多态性与非创伤性股骨头坏死发生有明显的关系.     

华东地区汉族群体D10S2325基因座的遗传多态性研究

目的 研究D10S2325基因座等位基因和基因型频率在华东地区汉族群体的分布情况，评价该基因座在华东汉族人群的法医学应用价值。方法 用PCR扩增、非变性聚丙烯酰胺凝胶垂直电泳、银染显色方法检测D10S2325基因座的遗传多态性。结果 在华东地区汉族人群中共检出12个等位基因(6～17)，其CE值和DP值分别为0．628和0．948。结论 D10S2325等位基因频率分布好，提供的信息量大，对华东地区汉族人群具有较高的非父排除率和个人识别能力，是群体遗传学研究和法医学应用理想的STR位点。     

血管紧张素原基因与黎族原发性高血压并脑梗死的相关性(英文)

目的研究血管紧张素原基因(angiontensinogen,AGT)4个位点的单核苷酸多态及其构成的单倍型与海南黎族原发性高血压及原发性高血压并发脑梗死的相关性。方法选择海南黎族人群为研究对象,利用PCR直接测序法,在100例原发性高血压患者、100例原发性高血压并脑梗死患者和100例健康对照者中,对AGT基因启动子区域的-152(G-A),-20(A-C),-18(C-T),-6(A-G)多态进行基因分型。结果C-18T多态的基因型和T等位基因频率分布在原发性高血压组(P=0.003,P=0.004)和原发性高血压并脑梗死组(P=0.002,P=0.002)均比健康对照组显著增高。-6(A-G)多态的G等位基因频率在原发性高血压并脑梗死组显著高于健康对照组(P=0.016),而与原发性高血压组比较无显著性差异。单倍型分析提示,由-20C和-18T构成的H5单倍型在原发性高血压组中增加,与对照组相比有显著性差异(P=0.006);而由-20C和-6G构成的H6单倍型在原发性高血压并脑梗死组中明显增加,与对照组相比有显著性差异(P=0.003)。结论在海南黎族人群中,AGT基因的-20(A-C),-18(C-T)多态可能对原发性高血压的发病起了重要作用;-20(A-C),-18(C-T)和-6(A-G)多态可能是原发性高血压并脑梗死发病的遗传危险因素。     

IL-1Ra基因多态性在冠心病病人心血管事件中的作用

目的 探讨白细胞介素-1受体拮抗剂(interleukin-1 receptor antagonist,IL-1Ra)基因多态性与冠心病患病率及心血管事件发生的关系.方法 利用聚合酶链式反应(PCR)技术对220例冠心病患者及100例正常对照者IL-1Ra基因进行扩增;检测冠心病患者的高敏感性C反应蛋白、血清总胆固醇、甘油三酯、低密度脂蛋白和体重指数;所有冠心病患者随访12个月,了解患者在出院后1年内的心血管事件.结果 冠心病患者和正常人的IL-1Ra基因多态性分布的差别没有统计学意义;出院后未发生心血管事件的冠心病患者Ⅱ型基因型携带率高于其他基因型.结论 监测IL-1Ra基因多态性不能预测冠心病的发生;但其多态性中Ⅱ型基因对已患冠心病患者的预后有一定保护作用.