SCN7A 基因单核苷酸多态性与原发性与原发性高血压的相关研究

目的　检测中国人电压调控钠通道 7型α亚单位基因 (sodium channel,voltage- gated,type ,alpha polypeptide,SCN7A)调控区和编码区的单核苷酸多态性 (single nucleotide polymorphisms,SNPs) ,并探讨其与上海汉族人群原发性高血压的关系。　方法　采用直接测序法检测基因启动子、编码区和部分内含子的序列 ,以确定中国人群中 SCN7A基因 SNPs的位置及类型。采用聚合酶链反应 -限制性片段长度多态性及直接测序法 ,对上海汉族 96例原发性高血压患者和 96名正常血压对照者进行 SNP检测和关联研究。对所发现的 P<0 .0 5的 SNP位点 ,进一步扩大样本 (病例、对照组各 2 88例 )加以验证。结果　在 13132 bp的测序长度中 ,共发现 32个 SNP,包括启动子区 7个 ,编码区 10个 (其中改变氨基酸编码的 6个 ) ,3′非编码区 1个 ,内含子区 14个 ,其中 30个为新发现的 SNP。关联研究结果显示 SNP0 2 1在病例和对照组中的分布差异存在显著性 (P <0 .0 1) ,该 SNP多态可改变氨基酸的编码序列。　结论SCN7A基因变异可能与上海汉族人群原发性高血压相关。     

广西人群TLR4基因3′-非编码区单核苷酸多态性的研究

目的通过对TLR4(Toll-like receptor 4)基因3′-非编码区单核苷酸多态性(single nucleotide polymorphism,SNP)基因型和等位基因的研究分析,了解其在广西地区人群的频率分布及其在不同地区、种族之间分布是否存在差异。方法采用DNA测序方法和单碱基延伸PCR技术检测广西地区人群TLR4基因3′-非编码区rs11536889G/C多态性,并比较广西地区人群与人类基因组计划研究的4个人群(欧洲、中国北京、日本、非洲)(http://www.ncbi.nlm.nih.gov/projects/SNP/)以及韩国地区人群SNP分型数据,以分析TLR4基因rs11536889G/C基因型和等位基因在不同地区人群中的分布差异。结果广西地区人群的TLR4基因3′-非编码区rs11536889G/C位点的分布频率在男性、女性组间无差异(P0.05)。与日本、韩国、中国北京地区人群比较分布差异无统计学意义(P0.05);而与欧洲、非洲地区人群比较,分布有显著性差异(P0.05)。结论广西地区人群TLR4基因3′-非编码区单核苷酸多态性与其它地区、种族人群之间存在统计学差异。     

湖北汉族人群Tim-3基因启动子及编码区的分子突变

目的：检测湖北汉族人群Tim-3基因启动子区和编码区的单核苷酸多态性，寻找Tim-3基因的遗传标记。方法：采用分段扩增直接测序的方法检测60名湖北汉族人Tim-3基因的启动子区、全部的外显子区及部分内含子区，将测序结果与NCBI及HapMap计划库中其他人种的数据进行对比，确定湖北汉族人群Tim-3基因突变的位置、类型和频率。结果：在Tim-3基因启动子区和外显子区共发现9个SNPs，包含5个已报道的SNPs和4个新发现的突变位点。湖北汉族人群中检出的4个SNPsrs4704853、rs10515746、rs4704846、rs9313439与Ft本人分布相似（P〉0．05），与欧洲人及非洲人的分布则有统计学意义（P〈0．01）。结论：湖北汉族人群Tim-3基因的SNPs分布有别于其他人种，可为在汉族人群中研究Tim-3基因与疾病关联提供依据。     

中国人群IL-1R1基因变异及其对跨膜区功能影响的预测

目的　检测中国人白细胞介素 1Ⅰ型受体基因 (interleukin 1receptortypeⅠ ,IL 1)调控区和编码区的单核苷酸多态性 (singlenucleotidepolymorphisms ,SNPs) ,并初步探讨其可能对中国人IL 1R1的功能影响。方法　采用直接测序的方法检测基因的 5′区、编码区、部分内含子区和 3′区 ,以确定中国人群中IL 1R1基因SNP的位置和类型 ,用生物信息学方法对编码区SNP的功能进行了预测。结果　在 964 3bp的测序长度中 ,共发现了 16个SNP ,包括 5′区 4个 ,内含子区 4个 ,编码区 1个 ,3′非编码区 7个。其中一个SNP能引起IL 1R1跨膜区氨基酸的替代 ,生物信息学分析显示它能够引起跨膜区结构的改变。结论IL 1R1基因变异可能对IL 1R1的功能产生影响。     

广东汉族正常人群TLR4基因单核苷酸多态性研究(英)

目的：人类Toll样受体4（TLR4）是先天免疫系统中一个重要的病原微生物识别受体。本研究将建立中国汉族正常人群TLR4基因座位的单核苷酸多态性图谱。方法：收集191例健康、无亲缘关系的中国广东汉族人外周血液,通过对TLR4基因的启动子区、3个外显子区以及它们周围的内含子区进行PCR扩增和测序,得到汉族正常人群TLR4基因座位单核苷酸多态性图谱及其频率分布特点。结果：共发现8个单核苷酸多态性位点,其中5个是首次发现的新位点。分布频率最高（0.283）的单核苷酸多态性位点是-1607 C/T。常见于高加索人中的2个非同义突变Asp299Gly和Thr399Ile在汉族人中没有被发现。中性检验显示汉族人群TLR4基因符合中性进化模型。结论：本研究建立了汉族正常人群TLR4基因座位的单核苷酸多态性图谱,发现了一些种族特异性的单核苷酸多态性位点,这些工作将为今后开展汉族人基因多态性与疾病相关性研究以及人群进化研究提供一定的帮助。     

抗肌萎缩蛋白基因内MPIP位点多态性的银染法检测

应用银染聚丙烯酰胺凝脏法．对抗肌萎缩蛋白基因3′非翻译区MPIP位点进行了扩增片段长度多态性分析。此位点在中国人中有多态性，共检出3个等位片段，汉族人群中3个等位片段的频率分别为0.141、0．703、0．156，PIC为0．462；回族人群中的频率分别为0．182．0．591．0．227，PIC为0．566。此位点与3′不翻译区的3′CA位点存在连锁不平衡性．在连锁分析中可替代3′CA作为基因3′端的遗传标志。     

广东汉族人群TLR2基因的多态性研究

目的:人类Toll样受体2(TLR2)是先天免疫系统中一个重要的病原微生物识别受体。本研究将建立广东汉族人群TLR2基因座位的功能性多态性图谱,为下一步疾病相关性研究打下基础。方法:收集200例健康、无亲缘关系的中国广东汉族人外周血液,随机抽取其中24例样品,对TLR2基因的启动子区、3个外显子以及它们周围的部分内含子序列进行聚合酶链式反应(PCR)扩增和直接测序,找出多态性位点,对剩余176例样品分别用序列特异性引物聚合酶链反应(PCR-SSP)及PCR技术对发现的单核苷酸多态性(SNPs)和插入/缺失(INDEL)多态性位点进行基因分型,分型结果进行Hardy-W e inberg平衡分析、中性进化分析以及连锁不平衡分析。结果:发现5个SNPs位点,其中2个位于启动子区的SNPs是首次发现,位于编码区的3个SNPs位点均为同义突变,频率最高的SNP是rs3804099,其次要等位基因频率为26.3%;在第1外显子区发现1个长度为22bp的INDEL多态位点(-196到-174),其缺失等位基因所占的频率为31.8%。所有多态性位点均符合Hardy-W e inberg平衡。中性检验显示广东汉族人群TLR2基因符合中性进化假说。连锁不平衡分析显示位于调控区的-18945 C/T和-18883 C/G 2位点之间完全连锁,而位于编码区的rs3804099和rs3804100两位点之间紧密连锁。结论:本研究首次建立了汉族正常人群TLR2基因座位的功能性多态性图谱,并研究了其分布频率,发现了一些种族特异性的多态性位点,为今后开展汉族人基因多态性与疾病相关性研究以及人群进化研究提供了重要资料。     

浙江地区汉族人群caspase-3基因三个位点的单倍型研究

目的分析汉族人群caspase-3基因的单核苷酸多态性(singlenucleotidepolymorphisms,SNPs)位点及其构成的单倍型,为研究caspase-3基因对凋亡调节机制的个体差异提供线索。方法用变性高效液相色谱技术和DNA测序技术检测caspase-3基因的调控区、第2～7外显子及部分侧翼序列的多态性位点;分析位点间的连锁不平衡关系,估算它们构成的单倍型。结果共检出3个SNP位点(C829A、A17532C、C20541T),分别位于caspase-3基因的5′端调控区、第4内含子和3′调控区;3个位点间存在强连锁不平衡,其中位点A17532C与C20541T呈完全连锁不平衡;54.3%的C-829/A-17532/C-20541是汉族人群的主要单倍型。结论浙江地区汉族人群caspase-3基因上的3个SNP位点间存在强连锁不平衡,它们构成的主要单倍型不同于北美人群。     

中国汉族人CC趋化因子受体-5基因新的突变位点及多态性分析

目的：调查中国汉族人群人类免疫缺陷病毒－1协同受体CC趋化因子受体-5[chemokine (CC)receptor 5,CCR5]编码区的基因多态性位点，为艾滋病的防治提供依据。方法：CCR5编码区用两对引物进行PCR扩增，设计测序引物依次测序，样本数为42份，用DNAstar分析测序结果，寻找单核苷酸多态性（single nucleotide polymorphism,SNP)位点。结果：在编码区共发现6个SNP位点，4个引起氨基酸改变：A184G、G503T、G668A、G999T；1个单碱基缺失，引起移码突变和提前终止。A184G、G503T、G999T3个中国汉族人所特有的SNP位点为首次发现，等位基因频率分别为1．2％，39．0％和9．5％；其中G503T分布明显不符合Hardy-Weinberg平衡。结论：中国汉族人CCR5编码区SNP位点有自己的特点，与高加索人和非洲人明显不同，与日本人也不完全一致。     

中国北方汉族人群FUT5基因编码区序列特征

目的 揭示中国北方汉族人群岩藻糖基转移酶基因V(fucosyltransferase 5 gene,FUT5)编码区序列特征.方法 对160名健康中国北方汉族人群血液样品进行研究,DNA测序分析其中30例FUT5基因编码区序列;突变基因亚克隆后测序鉴定其单倍型;聚合酶链反应-限制性片段长度法分析130名C560T(rs778970)和C484A位点的遗传多态性.结果 DNA测序共鉴定出7个单核苷酸多态性(single nucleotide polymorphism,SNPs)及2个新突变点C484A(Leu162Met)和T684C;9种碱基替换共鉴定出7种单倍型;160名样本的rs778970位点遗传多态性分析结果显示等位基因频率C为0.3031,T为0.6969;而C484A未发现其具有多态性.结论 中国北方汉族人群FUT5基因编码区序列呈现出高度的变异性;rs778970位点等位基因分布具有较好多态性.     

广东地区汉族人群TLR1 基因多态性的测序研究

目的: 人类Toll样受体1(TLR1)在先天性免疫中起着重要作用。本文将着重研究广东地区汉族正常人群中TLR1 基因功能区的单核苷酸多态性(SNPs)图谱和频率分布。方法: 随机收集50例健康、无亲缘关系的中国广东地区汉族人外周血液,对TLR1 基因的启动子区、5'和3'非翻译区、4个外显子区的序列进行PCR扩增和直接测序,找出多态性位点及其频率分布规律。在此基础上对多态性位点进行Hardy-Weinberg平衡分析、中性进化分析和连锁不平衡分析。结果: 共发现17个SNPs以及2个插入/缺失多态位点,其中2个是首次发现的新多态性位点。位于编码区的新SNP位点+1 378 A/G为非同义突变位点,能导致460位丝氨酸(Ser)残基替换为甘氨酸(Gly)残基,并且这个氨基酸残基的替换处于TLR1胞外区的LRR结构域中,从而有可能影响蛋白的识别功能。另外,频率最高的SNPs是+743 A/G和+1 518 A/G,其次要等位基因频率均达到48%。所有多态性位点均符合Hardy-Weinberg平衡。中性检验显示广东汉族人群 TLR1 基因不符合中性进化假说,很可能是其调控区受到平衡选择作用的原因。连锁不平衡分析显示多态性位点-6 912 C/TA、-6 876 C/T、-6 399 C/T和-6 375 C/T之间,-6 847 A/G和-6 737 A/T之间,以及-5 984 -/CT、-5 531 A/G和-5 490 C/G之间完全连锁。结论: 本研究首次报道了汉族正常人群TLR1 基因的功能性多态性图谱,发现了一些种族特异性的多态性位点及频率分布规律,为今后开展汉族人基因多态性与疾病相关性研究打下一定的基础。     

Toll-like receptor gene polymorphisms are associated with allergic rhinitis: a case control study

ABSTRACT: BACKGROUND: The Toll-like receptor proteins are important in host defense and initiation of the innate and adaptive immune responses. A number of studies have identified associations between genetic variation in the Toll-like receptor genes and allergic disorders such as asthma and allergic rhinitis. The present study aim to search for genetic variation associated with allergic rhinitis in the Toll-like receptor genes. METHODS: A first association analysis genotyped 73 SNPs in 182 cases and 378 controls from a Swedish population. Based on these results an additional 24 SNPs were analyzed in one Swedish population with 352 cases and 709 controls and one Chinese population with 948 cases and 580 controls. RESULTS: The first association analysis identified 4 allergic rhinitis-associated SNPs in the TLR7-TLR8 gene region. Subsequent analysis of 24 SNPs from this region identified 7 and 5 significant SNPs from the Swedish and Chinese populations, respectively. The corresponding riskassociated haplotypes are significant after Bonferroni correction and are the most common haplotypes in both populations. The associations are primarily detected in females in the Swedish population, whereas it is seen in males in the Chinese population. Further independent support for the involvement of this region in allergic rhinitis was obtained from quantitative skin prick test data generated in both populations. CONCLUSIONS: Haplotypes in the TLR7-TLR8 gene region were associated with allergic rhinitis in one Swedish and one Chinese population. Since this region has earlier been associated with asthma and allergic rhinitis in a Danish linkage study this speaks strongly in favour of this region being truly involved in the development of this disease.     

No association of toll-like receptor 2 polymorphisms with Alzheimer's disease in Han Chinese

Toll-like receptor 2 (TLR2) represents a reasonable functional and positional candidate gene for Alzheimer's disease (AD) as it is located under the linkage region of AD on chromosome 4q, and is functionally involved in the microglia-mediated inflammatory response and amyloid β (Aβ) clearance. In the current study, 7 single nucleotide polymorphisms (SNPs) that span the TLR2 were selected and their associations with late-onset AD (LOAD) risk were assessed in a case-control sample comprising 785 individuals in a Han Chinese population. No significant differences in the frequency of TLR2 alleles, genotypes, and haplotypes in the AD cases were detected compared with the controls. TLR2 gene might not play a major role in the genetic predisposition to late-onset Alzheimer's disease in this population.     

SNPs in the exons of Toll-like receptors are associated with susceptibility to type 1 diabetes in Chinese population

Objective

Toll-like receptors (TLRs) recognize a wide range of pathogen-associated molecular patterns (PAMP) and mount the initiation of immune response. Single nucleotide polymorphisms (SNPs) in exons of genes encoding TLRs might be responsible for the generation of an abnormal immune response which could lead to autoimmune diseases. In this study, we investigated the SNPs in TLRs in a Chinese population, and we hypothesized that SNPs in TLRs are associated with type 1 diabetes (T1D), an autoimmune disease caused by destruction of insulin producing pancreatic β-cells, in the studied population.

Research design and methods

We selected 28 SNPs in exons of TLRs with an aim to identify those that might have a direct correlation with T1D etiology and many have not been included in previous GWAS studies. Genotyping of those SNPs in TLRs was performed in 429 T1D patients and 300 age and gender-matched healthy controls in Chinese Han population which was not included in the earlier GWAS studies.

Results

Among the SNPs genotyped, the T allele of TLR1–626 was positively associated with T1D (OR = 1.98, Pc = 0.01). We identified another T1D association locus in TLR6: the homozygous AA genotype of TLR6-1329 was negatively and heterozygous GA was positively associated with T1D (OR = 0.54, Pc = 0.02 and OR = 1.70, Pc = 0.03, respectively). We also identified the haplotype T-G-A in TLR1 gene to be positively associated with T1D (OR = 2.22, Pc = 0.03). Additional haplotypes in TLR-6 also showed significant positive and negative association. In addition, our haplotype analysis and conditional analysis showed that these two SNPs are the primary T1D associated loci among the SNPs tested in our cohort in each TLR gene.

Conclusion

SNPs and haplotypes in TLR1 and TLR6 gene were associated with T1D in Chinese Han population. Our study, for the first time, indicates that TLR1 and TLR6 gene might play important roles in the etiology of T1D.     

Clinical relevance of tag single nucleotide polymorphisms within the CAT gene in patients with PTSD in the Chongqing Han population

Background: Free radical-induced oxidative damage of the brain has been implicated in a number of psychiatric disorders, including post-traumatic stress disorder (PTSD). Catalase (CAT) is a major antioxidant enzyme and a number of polymorphisms in CAT have been shown to be associated with several diseases, including hypertension, diabetes mellitus, Alzheimer’s disease, and vitiligo. The aim of this study was to evaluate the association of CAT gene polymorphisms with PTSD in a case-control study. Materials and methods: A total of 460 unrelated adult Chinese Han adults, including 287 healthy volunteers and 173 patients with PTSD. Six tag single-nucleotide polymorphisms (tSNPs) were selected from the entire CAT gene through construction of haplotype bins, and they were genotyped using an improved multiplex ligation detection reaction (iMLDR) technique. Allelic frequencies and clinical characteristics were compared in two independent Chinese Han populations. Results: Six tag SNPs were identified in the Chinese Han population and all were common SNPs. However, we could detect no evidence of genetic association between six tag SNPs in the CAT gene and PTSD in the Chinese Han population. Conclusions: This result suggests that six tag SNPs of the CAT gene may not be associated with PTSD, and that CAT gene might not influence the development of PTSD in patients following exposure to a traumatic event, also may be the sample sizes too small to allow a meaningful test.     

新疆维吾尔族人STEAP4基因遗传变异的分布及其与代谢综合征的关联

目的 探讨前列腺六次跨膜上皮抗原4(six transmembrane epithelial antigen of prostate 4,STEAP4)基因多态与维吾尔族人代谢综合征(metabolic syndrome,MetS)的关联.方法 采取以流行病学调查为基础的病例对照研究,选择682例MetS和1228名对照为研究对象.测序筛查维吾尔族MetS患者STEAP4基因功能区的变异,选择代表性变异在维吾尔族自然人群中进行关联分析.结果 维吾尔族人STEAP4基因单核苷酸多态(single nucleotide polymorphism,SNP)的最小等位基因频率不同于中国北京汉族人和欧洲人.rs34741656多态与MetS相关[P=0.034,控制年龄、性别后OR=0.757(95%CI:0.584～0.982)],在对照组,该位点与空腹和餐后2 h血糖水平有关(分别P=0.049,P=0.027).常见单倍型H4(rs8122/rs1981529/rs34741656,G-A-A)可能与MetS有关(permutation P=0.089).结论 STEAP4基因遗传变异可能与维吾尔族人代谢综合征相关.

Abstract：

Objective To investigate the genetic variations of the six transmembrane epithelial antigen of prostate 4 gene (STEAP4)in Chinese Uygur patients with metabolic syndrome (MetS) and to analyze the association of the representative genetic variations of STEAP4 gene with MetS in the population.Methods The sequences of STEAP4 gene functional region (all exons, exon-intron boundaries and the putative promoter region, including the -1 kb 5' and 3'untranslated regions) were amplified and sequenced for patients with MetS. The representative variations were selected based on the function (missense mutation) and linkage disequilibrium (γ2 ＞0. 8) and genotyped with TaqMan-PCR method in 1910 general populations (682 MetS and 1228 non-MetS controls). The subjects were selected from the cross-sectional study of obesity, hypertension, diabetes, dyslipidemia from January to February 2007 among Uygur people,a relatively isolated population with a relatively homogeneous environment, in Hetian area in Xinjiang Uygur Autonomous Region. Results (1) Fourteen novel and six known single nucleotide polymorphisms (SNPs)or mutations, including 2 missense mutations, were identified at the functional region of STEAP4 gene in 96 Uygur patients with MetS. The minor allele frequencies of the SNPs of STEAP4 gene in Uygur population were different from that in European and Chinese Han in Beijing area. (2) The SNP 364G/A (rs34741656,Ala122Thr) was significantly associated with MetS [dominant model P = 0. 034, OR = 0. 757 (95% CI:0. 584-0. 982) adjusted for age and gender], and was associated with fasting blood glucose (FBG) (P =0. 049) and 2-hour postprandial glucose (2HPG) (P = 0. 027) levels in controls. In this SNP, the AA carriers had lower blood glucose levels compared with subjects carrying GG and GT genotypes. (3) The common haplotype H4 (rs8122/rs1981529/rs34741656, G-A-A), may be associated with MetS (permutation P= 0. 089). Conclusion STEAP4 genetic polymorphisms may be associated with MetS risk in Chinese Uygur population.