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。     

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。     

中国北方汉族人群中载脂蛋白M基因多态性分布特征及连锁不平衡分析

目的研究中国北方汉族人群中载脂蛋白M基因（apolipoprotein M,APOM）多态性分布特征及其连锁不平衡关系。方法采用PCR扩增基因组DNA直接测序法结合PCR-限制性片段长度多态方法对330名中国北方汉族健康人群的APOM基因单核苷酸多态性（single nucleotide polymorphism,SNP）进行分析。结果中国北方汉族人APOM基因存在1号内含子rs805264位点、5号内含子rs707922位点及rs707921位点3个多态位点,不同种族及地区APOM基因SNP差异有统计学意义。APOM基因rs805264位点、rs707922位点及rs707921位点SNP在中国北方汉族人群中呈现明显的连锁不平衡,主要有G-G-C、A-T-A两种单体型。结论APOM基因SNP在中国北方汉族人群中存在显著连锁不平衡。     

TLR7基因单核苷酸多态性与江苏淮安哮喘患者易感性的相关性研究

目的:探讨Toll样受体7(TLR7)基因rs179009、rs179019、rs5935436位点多态性与江苏淮安地区汉族人群支气管哮喘的相关性。方法:采用病例对照方法,以聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法比较158例支气管哮喘组与137例健康对照组之间基因型、等位基因频率的差异。结果:哮喘组TLR7基因rs179009、rs170019位点基因型及等位基因型频率与对照组相比差异有统计学意义(P<0.05)。rs5935436位点基因型及等位基因型频率在哮喘组和对照组间差异均无统计学意义(P>0.05)。结论:TLR7基因rs179009位点和rs179019位点的多态性可能与江苏淮安地区汉族人群哮喘相关;而rs5935436位点的多态性可能与江苏淮安地区汉族人群哮喘无关。     

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。     

新疆柯尔克孜族过氧化物酶体增殖物激活受体基因单核苷酸多态性

目的:分析柯尔克孜族健康人群过氧化物酶体增殖物激活受体基因(PPARG)的31个单核苷酸多态性(SNP)位点的遗传多态性;方法:利用Hap Map软件筛选31个SNPs位点,利用质谱检测技术进行多态性检测并根据质谱峰图判读样本目标位点基因型,利用χ~2检验确定筛选的SNP位点是否符合Hardy-Weinberg平衡定律并分析柯尔克孜族与其他民族间基因型和等位基因频率差异。结果:在31个SNP位点中,23个位点的最小等位基因频率MAF≥0.05具有多态性;在23个SNPs中rs1175540、rs17036242、rs2881654、rs2959273、rs2972162、rs4135275、rs709151、rs9310401、rs1801282位点在柯尔克孜族和维吾尔族人群间基因型和等位基因分布频率差异均有统计学意义;rs2292101、rs3856806、rs7626560位点在柯尔克孜族和北京汉族人群、犹他州居民、伊巴丹尼日利亚人群间基因型频率差异有统计学意义;rs3856806、rs4135275、rs6782475、rs7626560位点在柯尔克孜族和北京汉族人群、犹他州居民、伊巴丹尼日利亚人群间等位基因频率差异有统计学意义。结论:PPARG基因23个SNP位点多态性在新疆柯尔克孜人群和不同种族问差异具有统计学意义,这种差异可能是导致某些疾病在不同种族间的发现率和临床表现存在显著不同的因素之一。     

TLR4基因多态性在中国人群中的初步研究

目的检测中国人Toll样受体4(Toll—like receptor 4．TLR4)基因调控区和编码区的单核苷酸多态性(single nucleotide polymorphisms，SNPs)．寻找TLR4基因的遗传标记。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TLR4基因SNP的位置和类型，并用聚合酶链反应-限制性片段长度多态性对重庆汉族样本进行了抽样调查。结果在4．98kb的测序范围内，发现5个新的SNP，3个位于5′区．2个位于3′非翻译区。在重庆地区汉族样本中．两个高频分布SNP的等位基因频率分别是0．266和0．404。结论在TLR4基因新发现的两个高频多态性位点在我国人群中比较常见，可以作为关联分析的遗传标记。     

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。     

TBX21基因多态性与类风湿关节炎相关性的研究

目的:研究TBX21基因单核苷酸多态性(SNPs)与中国汉族人群类风湿性关节炎(RA)的关系。方法:采用单碱基延伸法(SBE)检测288例RA患者和288名正常健康者TBX21基因的5个SNPs:rs4794067、rs2240017、rs17250932、rs2074190和rs12721470的基因型。结果:5个SNP位点的基因型均符合Hardy-Weinberg平衡(P0.05)。rs12721470位点的基因型频率和等位基因频率在RA组和对照组间的差别具有统计学意义。rs4794067位点的基因型频率在RA组和对照组间无统计学差异,而等位基因频率在RA组和对照组间的差异则具有统计学意义(P0.05)。rs17250932、rs2240017和rs2074190基因型及等位基因频率在RA组和对照组间无统计学意义(P0.05)。结论:TBX21基因单核酸多态性rs12721470与中国汉人群类风湿关节炎是显著相关联的。     

广西壮族及汉族人群CD40配体基因rs3092923G/A和rs3092929A/C遗传多态性的研究

目的探讨CD40配体基因rs3092923G/A和rs3092929A/C多态性位点在广西壮族及汉族人群中的分布,同时比较其基因型及等位基因频率分布在不同种族人群之间以及同一种族不同性别之间存在的差异。方法采用单碱基延伸PCR的检测方法,分析201名广西汉族人和199名广西壮族人的CD40配体基因rs3092923G/A和rs3092929A/C多态性。结果在广西壮族人群中,CD40配体基因rs3092923G/A位点AA、AG与GG基因型频率和rs3092929A/C位点AA、AC与CC基因型频率均为86.4%、7.5%和6.0%,rs3092923G/A位点的A、G等位基因频率和rs3092929A/C位点的A、C等位基因频率均为90.2%、9.8%;在广西汉族人群中,CD40配体基因rs3092923G/A位点AA、AG与GG基因型频率和rs3092929A/C位点AA、AC与CC基因型频率均为93.0%、4.0%、3.0%,rs3092923G/A位点的A、G等位基因频率和rs3092929A/C位点的A、C等位基因频率均为95.0%、5.0%。将这2个多态性位点基因型分布频率在2个民族人群中比较,差异均无显著性(P均>0.05),而等位基因频率却有着显著性差异(P均<0.05)。另外,将这2个位点多态性分布频率在男女性别之间作比较,差异都没有显著性(P均>0.05)。进一步与人类基因组计划公布的4个人群相比,广西汉族人群的rs3092923G/A和rs3092929A/C 2位点基因型和等位基因频率与非洲、日本、欧洲和北京人群比较,差异都具有显著性(P均<0.05)。结论在广西地区壮族及汉族人群中存在着CD40配体基因多态性。广西汉族人群CD40配体基因多态性的分布频率同其他种族人群比较存在着显著性差异,这种差异可能是导致与CD40配体相关的疾病在不同种族人群间的临床表现以及发病率存在明显不同的原因之一。     

IL-1、IL-10及TNF-β单核苷酸多态性与福建泉州地区汉族人群类风湿性关节炎的相关性研究

目的:探讨福建泉州地区汉族人群类风湿性关节炎(RA)患者炎症相关因子的单核苷酸多态性(SNP)位点的分布特征,并研究其与RA易感性的关系。方法:选取福建泉州地区汉族人群RA患者155例(RA组)和健康体检者170例(对照组)为研究对象,采用全血三引物等位基因特异性扩增(TRIP-ASPCR)技术对RA相关的SNP位点进行基因分型。运用SPSS 19.0统计分析软件,采用卡方检验和Logistic法分析Hardy-Weinberg遗传平衡吻合度、SNP位点等位基因与RA发病关联强度,并利用SHEsis软件对所选SNP位点进行连锁不平衡分析和单倍型分析。结果:对照组中有1个SNP位点符合Hardy-Weinberg平衡检验(P>0.05),RA组有1个SNP位点符合Hardy-Weinberg平衡检验(P>0.05);在对照组和RA组之间有4个SNP位点的等位基因频率差异存在统计学意义(P<0.05,OR>1),与RA呈正相关,且相关性明显。结论:RA患者中,存在4个SNP位点(IL-10 rs1800893、IL-1βrs16944、TNF-βrs2009658和...     

α2-Heremans-Schmid糖蛋白基因多态性与其血清含量的关联研究

目的 探讨α2-Heremans-Schmid糖蛋白(α2-Heremans-Schmid glycoprotein,AHSG)基因单核苷酸多态(single nucleotide polymorphism,SNP)与其血清含量的相关性及其参与调节基因转录的机制.方法 对AHSG基因进行重测序,构建连锁不平衡模式,选择标签SNP在192名北京汉族和424名广州汉族个体中进行了基因分型;应用报告基因荧光素酶活性检测不同等位基因的转录活性;应用酶联免疫吸附试验检测北京地区192名个体血清中AHSG的浓度.结果 AHSG重测序共检出8个SNP,连锁不平衡分析显示广州和北京地区人群的连锁不平衡模式不同,但选择的标签SNP和具有潜在功能的SNP的等位基因和基因型频率分布在两组人群中差异无统计学意义.荧光素酶活性实验结果显示,AHSG基因启动子区-799A等位基因比-799T等位基因的转录活性高;多态性位点rs2248690、rs4917和rs4918与血清中AHSG的含量存在相关性;多元回归分析结果表明只有rs2248690与AHSG含量存在相关性.结论 位于AHSG基因启动子区的rs2248690位点与其血清中含量存在相关性.     

Novel exonic mu-opioid receptor gene (OPRM1) polymorphisms not associated with opioid dependence.

The mu-opioid receptor (MOR) mediates reward and dependence associated with opioids and other commonly abused substances. Variability in the MOR gene, OPRM1, may influence risk for opioid dependence. In this study, associations between two single nucleotide polymorphisms (SNPs), dbSNP rs540825 and dbSNP rs562859, and opioid dependence were investigated. The two SNPs are located in the protein coding region of the novel exon X of an alternative splice variant of OPRM1, and can be detected using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. Genotyping at the two SNPs was performed for 170 severe opioid dependent individuals and 128 carefully screened controls. Although no differences were found between cases and controls, there were significant prevalence differences between African-American (AA) subjects and European-American (EA) subjects for SNP 540825 allele and genotype frequencies. The 540825 and 562859 polymorphisms were found to be in complete linkage disequilibrium (LD) for both ethnic groups, and LD existed between the 562859 SNP and the A(-1320)G SNP in the promoter region of OPRM1 in AAs, based on genotyping data previously carried out on the same subjects. LD between these two markers, separated by 55 kb, links the entire distance studied in this project. The results indicate that polymorphisms in the novel splice variant are not associated with opioid dependence, but are in LD with other polymorphisms in OPRM1.     

DVL2基因多态性与中国汉族人群先天性脊柱侧凸遗传易感性的关联研究

背景：近20年来小鼠的分子胚胎学研究进展获得了大量关于脊椎发育的分子信息,用同线性分析法确立先天性脊柱侧凸的候选基因已成为可能。 目的：通过候选基因DVL2上关键单核苷酸多态性位点的筛查,探索DVL2与中国汉族人群先天性脊柱侧凸及其不同临床表型之间的关联。 方法：采用病例-对照研究,入选127例中国汉族先天性脊柱侧凸患者和127例对照组。根据国际人类基因组单体型图计划提供的基因型数据,应用Haploview 4.1软件选取DVL2的标签和功能单核苷酸多态性。根据椎体畸形特点、畸形部位、畸形受累程度、有无合并肋骨畸形和椎管内畸形将病例组进一步分为不同临床表型。对所有样本应用SNPstream UHT Genotyping系统对所选单核苷酸多态性位点进行基因型鉴定;进一步进行基于基因型/等位基因频率的关联分析,并用Haploview 4.1软件分析对照组单核苷酸多态性位点间是否存在连锁不平衡。 结果与结论：共筛选5个位点：单核苷酸多态性1(rs2074222)、单核苷酸多态性2(rs222837)、单核苷酸多态性3(rs222835)、单核苷酸多态性4(rs10671352)和单核苷酸多态性5(rs222836),其基因型分布在病例和对照组中均符合Hardy-Weinberg平衡;5个位点处于完全连锁不平衡状态;5个位点的基因型/等位基因/单倍体型与先天性脊柱侧凸的发生风险之间不存在相关性。在进一步与先天性脊柱侧凸临床表型的关联分析中没有发现阳性位点。提示在中国汉族人群中DVL2基因可能不是引起先天性脊柱侧凸及其不同临床表型的主要因素,有待于进一步深入研究。     

Positive association of the <Emphasis Type="Italic">FTSJ1</Emphasis> gene polymorphisms with nonsyndromic X-linked mental retardation in young Chinese male subjects

To investigate the possible genetic association of nonsyndromic X-linked mental retardation (NS-XLMR) with FTSJ1 gene polymorphisms, a case-control association study was performed focusing on the Chinese Han population in the Qinba mountain region. Three common single nucleotide polymorphisms (SNPs) (rs2268954, rs2070991, rs5905692) in the gene were selected and genotyped using the polymerase chain reaction single-strand confirmation polymorphism (PCR-SSCP) method. Pairwise linkage disequilibrium (LD) analysis showed that the three SNPs were in strong LD (all D' > 0.8). There were significant differences between cases and controls in allele frequency distribution of rs2268954 (P = 0.036), rs2070991 (P = 0.043), and rs5905692 (P = 0.014) and in the distributions of common haplotypes combined by these SNPs (global P = 0.01236) in male subjects. In female subjects, however, no positive results were found. Our results suggest a positive association between the genetic variants of the FTSJ1 gene and NS-XLMR in young male subjects in the Chinese Han population in the Qinba region.     

Toll- like receptor 2 polymorphism and IL-6 profile in relation to disease progression in chronic HBV infection: a case control study in Egyptian patients

Chronic hepatitis B (CHB) has a wide range of outcomes depending on host immune responses mainly Toll-like receptors (TLRs) signaling and released cytokines. Toll-like receptor 2 (TLR2) single nucleotide polymorphisms (SNPs) and interleukin 6 (IL-6) may influence the course of CHB. We aimed to elucidate the relation between TLR-2 polymorphism, IL-6 profile, and CHB progression. We analyzed TLR-2 polymorphism (SNP; rs3804099) in 185 CHB patients and 60 controls using TaqMan allelic discrimination assay. Serum IL-6 levels were assessed by ELISA. IL-6 levels were considerably higher in active CHB and cirrhotic patients compared with inactive carriers and controls (P < 0.001). IL-6 showed positive correlation with ALT and advanced fibrosis in active CHB patients (r = 0.31, P = 0.02). A significant positive correlation was noticed between IL-6 and HBV DNA PCR in all CHB groups. TT genotype of rs3804099/TLR-2 was significantly more prevalent in inactive carriers compared to active hepatitis patients (P = 0.04, OR = 0.39 and 95% CI: 0.16–0.95). Both heterozygous CT and mutant TT genotypes were significantly more frequent among inactive carriers compared to cirrhotic patients (P = 0.01, OR = 0.33, 95% CI: 0.13–0.81 and P = 0.009, OR = 0.32, 95% CI: 0.13–0.77). TT genotype was significantly related to lower IL-6 levels in active hepatitis and cirrhotic groups (P = 0.005 and P = 0.001, respectively) showing that TLR mutations would be associated with milder hepatitis activity and lower possibility for disease progression. There may be a positive association between TLR2 rs3804099 polymorphism and hepatitis B activity. IL-6 is a good indicator of CHB disease progression.

     

Confirmation of an association between single nucleotide polymorphisms in the VDR gene with respiratory syncytial virus related disease in South African children

Respiratory syncytial virus is a leading cause of lower respiratory tract infection in infants. Disease severity has been linked to host immune responses and polymorphisms in genes associated with innate immunity. A large‐scale genetics study of single nucleotide polymorphisms (SNPs) in children in the Netherlands identified SNPs in the vitamin D receptor (VDR) and JUN genes which have a strong association with an increased risk of developing bronchiolitis following the first respiratory syncytial virus (RSV) infection. The Toll‐like receptor 4 (TLR4) gene has two SNPs which have been associated previously with RSV disease severity in various populations. The aim of this study was to determine if these SNPs may be associated with RSV disease in African children in South Africa. RSV patient (n = 296) and control (n = 113) groups were established (median ages: 3 and 3.5 months) and DNA extracted from the collected specimens. Real‐time polymerase chain reaction using hydrolysis probes was used to screen for SNPs in the VDR (Thr1Meth; rs10735810), TLR4 (Asp299Gly; rs4986790 and Thr399Ile; rs4986791) and JUN (c.750G/A; rs11688) genes. Carriers of the VDR (Thr1Meth) SNP minor T allele were more prone to RSV disease than individuals in the control group. The TLR4 (Asp299Gly), TLR4 (Thr399Ile), and JUN (c.750G/A) SNPs showed no significant association with RSV disease. It is concluded that children carrying the minor T allele of the VDR (Thr1Meth) SNP may be predisposed to RSV disease, as this SNP was identified as a risk factor for severe RSV disease in South African children, confirming the findings in the Netherlands. J. Med. Virol. 83:1834–1840, 2011. © 2011 Wiley‐Liss, Inc.