中国蒙古族人MBL基因启动子区SNP的研究

目的 研究中国蒙古族人群甘露聚糖结合凝集素(MBL)基因启动子区单核苷酸多态性(SNP)。方法 抽提人外周血白细胞基因组DNA，建立SSP-PCR及分子灯塔实时荧光PCR技术，检测MBL基因启动子区SNP位点-550(G／C，称H/L等位基因)、-220(G／C，X／Y等位基因)和 4(C／T，P／Q等位基因)，分析其单倍型及基因型频率。结果 从82人中检出等位基因型LYP／LYP4例(4．9％)，HYP／LYQ5例(6．1％)，LYP／LYQ35例(42．9％)，LXP／LXP1例(1．2％)，LYQ／LYQ11例(13．4％)，LXP／LYQ14例(17．1％)，HYP／LYP2例(2．4％)，HYP／LXP1例(1．2％)，HYP／HYP9例(11．0％)。结论 中国蒙古族人群MBL基因启动子区SNP等位基因型以LYP／LYQ、LXP／LYQ、LYQ／LYQ和HYP／HYP为主。     

中国维吾尔族人群MBL基因启动子及第一外显子区SNP及其单倍型与基因型研究

收集新疆维吾尔族自治区维吾尔族一般人群血标本,提取白细胞基因组DNA,以序列特异性引物-多聚酶链反应技术检测其甘露聚糖结合凝集素(MBL)基因启动子区单核苷酸多态性位点-550G/C(称H/L等位基因)、-221C/G(X/Y)、+4C/T(P/Q)和结构基因第一外显子点突变CGT52TGT、GGC54GAC和GGA57GAA(分别称为D、B、C等位基因,野生型即A等位基因),并分析其单倍型与基因型。发现MBL基因启动子区等位基因主要为L、Y、P,第一外显子等位基因只发现B,未检出C和D;检出5种单倍型,其频率分别是HYPA 0.282、LYPA 0.268、LXPA 0.260、LYPB 0.120、LYQA 0.070。检出12种基因型,其频率分别为HYPA/HYPA 0.183、LXPA/LXPA 0.141、LYPA/LYQA 0.113、LYPA/LYPA 0.112、LYPA/LXPA 0.085、HYPA/LYPA 0.085、LXPA/LYPB 0.085、HYPA/LXPA 0.070、HYPA/LYPB 0.042、LYPA/LYPB 0.028、LYPB/LYQA 0.028、YPB/LYPB 0.028。     

中国白族人群MBL基因SNP及其单倍型与基因型的研究

目的：研究中国云南白族人甘露聚糖结合凝集素（MBL）基因单核苷酸多态性（SNP）及其单倍型与基因型。方法：对MBL基因启动子区SNP位点-550G／C（称H／L等位基因）、-221C／G（X／Y）、＋4C／T（P／Q）已明确的白族DNA样本，采用序列特异性引物．多聚酶链反应技术检测结构基因第一外显子点突变CGT52TGT、CCC54GAC和CCA57CAA（分别称为D、B、C等位基因，野生型即A），并分析MBL基因的单倍型与基因型。结果：只检出GGC54GAC点突变，其频率为0．100；检出的5种单倍型及其频率是：HYPA0．250、LXPA0．107、LYQA0．407、LYPA0．135、LYPB0．100；各基因型及其频率为：LYPA／LYPA0．043、LXPA／LYQA0．143、LYPA／LYPB0．014、HYPA／LYQA0．086、LYPA／LYQA0．157、HYPA／LYPA0．014、LYPB／LYQA0．143、HYPA／LYPB0．043、LXPA／LXPA0．014、HYPA／LXPA0．043、LYQA／LYQA0．143、HYPA／HYPA0．157。结论：中国白族人群MBL基因存在GGC54GAC点突变，单倍型以LYQA和HYPA为主，基因型则多见LYPA／LYQA、HYPA／HYPA、LX—PA／LYQA、LYPB／LYQA和LYQA／LYQA。     

中国佤族人群MBL基因SNP及其单倍型与基因型的研究

甘露聚糖结合凝集素（mannan-binding lectin，MBL）通过激活补体凝集素途径和调理吞噬作用清除病原体及受感染细胞，在机体天然免疫中起关键作用。已发现MBL基因上存在至少20个单核苷酸多态性（SNP）位点，仅其中6个SNP位点对MBL血清水平有较大的影响，     

甘露聚糖结合凝集素基因启动子区多态性及其意义

甘露聚糖结合凝集素是机体重要的天然免疫防御分子，其血清水平主要受结构基因点突变的影响和启动子区多态性的调控。本文简介其启动子区基因结构、多态性及生物学意义。     

甘露聚糖结合凝集素基因启动子区多态性及其意义

甘露聚糖结合凝集素是机体重要的天然免疫防御分子 ,其血清水平主要受结构基因点突变的影响和启动子区多态性的调控。本文简介其启动子区基因结构、多态性及生物学意义。     

096 甘露聚糖结合凝集素基因启动子区多态性及其意义

甘露聚糖结合凝集素是机体重要的天然免疫防御分子，其血清水平主要受结构基因点突变的影响和启动子区多态性的调控。本文简介其启动子区基因结构、多态性及生物学意义。     

汉族儿童MBL基因启动子-221位点、外显子I区SNP和基因单体型分析

研究浙江省汉族儿童MBL基因启动子-221位点和外显子I区单核苷酸多态性(SNP)、基因单体型与血浆MBL水平的关系。血浆MBL浓度的检测采用ELISA法,MBL基因SNP分析采用序列分析法,基因单体型分析采用SHEsis软件,统计分析采用SPSS软件11.0版。结果105例汉族儿童中MBL基因启动子区-221位点X/Y和Y/Y基因型分别占19%和81%,该位点变异频率为0.095;外显子I基因型A/A、A/B和B/B分别占69.5%、27.6%和2.9%,B型变异频率为0.167。基因单体型有YA、YB、XA三种,其中以YA最多见,频率为0.741,其次是YB,频率为0.164。血浆MBL浓度范围为3~6 025 ng/ml,中位数为1057 ng/ml,外显子A/A型的MBL浓度显著高于A/B型,后者又显著高于B/B型;而启动子Y/Y型的MBL浓度比X/Y型高。本研究中汉族儿童MBL基因外显子I区+230位点的变异频率为0.167,外显子I区+230位点变异可导致血浆MBL水平明显下降。     

蒙古族与回族人群MBL结构基因型及其与血浆蛋白浓度的关系

目的：分析内蒙古自治区蒙古族人群和宁夏回族自治区回族人群血浆甘露聚糖结合凝集素（MBL）的结构基因型,并探讨其与血浆蛋白浓度的关系。方法：用ELISA法检测血浆MBL浓度,用序列分析法分析MBL基因外显子1区52、54和57密码子的单核苷酸多态性;统计分析采用SPSS软件11.0版,遗传学分析采用SHEsis软件。结果：79例蒙古族人和69例回族人血浆MBL浓度中位数分别为1686和1909ng/ml,两组间无显著性差异（Z=0.63,P=0.4）。蒙古族人群外显子1A/A型、A/B型和B/B型分别占62.0%、35.4%和2.5%,而回族人群A/A型、A/B型和B/B型分别占58.0%、30.4%和11.6%,蒙古族和回族人群＋230位点变异频率分别为0.203和0.268,两者变异频率无显著性差异（χ^2 1.772,P=0.183）。所有样本无C或D型外显子发现。A/A型外显子的血浆MBL浓度显著高于A/B型,后者又显著高于B/B型（χ^2 86.526,P〈0.01）。结论：蒙古族和回族人群MBL基因只有A/A,A/B和B/B3种外显子基因型,A/A型结构基因MBL浓度显著高于A/B型,又显著高于B/B型。     

系统性红斑狼疮患者MBL基因启动子区多态性的研究

探讨系统性红斑狼疮(SLE)患者甘露糖结合凝集素(MBL)基因启动子区-550(H/L)和-221(X/Y)位点多态性及其与血清MBL含量的关系.用序列特异性引物聚合酶链(SSP-PCR)法分析SLE组62例和正常组54名MBL基因启动子区-550(H/L)和-221(X/Y)位点多态性,同时用ELISA检测血清MBL水平.两组均存在HY、LY、LX单倍型,SLE组单倍型LX显著高于对照组(OR 2.23;95%CI 1.09～4.55;P=0.02);SLE组HY/HY基因型MBL水平显著高于其他基因型患者的MBL水平(LY/LY,LX/LX,HY/LY,HY/LX,LY/LX,P＜0.05);SLE组和对照组MBL含量低于1000μg/L者分别为40.32%和14.81%,两组相比有显著性差异(OR 3.89;95%CI 1.57～9.62;P=0.002).SLE组的血清MBL含量明显降低,与基因启动子区单倍型LX增高相关.     

幼年特发性关节炎患儿甘露糖结合凝集素基因启动子区SNP研究

目的探讨甘露糖结合凝集素(MBL)基因启动子区单核苷酸多态性(SNP)与幼年特发性关节炎(JIA)易感性的关系。方法对50例JIA患儿和48名正常健康儿童MBL基因启动子区SNP位点-550(G/C,称H/L等位基因)和-221(G/C,称X/Y等位基因)采用等位基因特异性PCR法(PCR-SSP)检测,并分析其单元型及基因型频率。结果共检出HY、LY和LX三种单元型,在JIA患儿中的频率依次为0.540、0.270和0.190,而在正常儿童中频率分别为0.594、0.292和0.114;两组间各单元型比较均无显著性差异。结论MBL基因启动子区单核苷酸多态性与JIA无相关性。     

Toll样受体9基因启动子单核苷酸多态性与儿童变应性哮喘的相关性

Objective To investigate the distribution characteristics of the single nucleotide polymorphisms (SNPs) in the promoter region of the toll-like receptor 9 gene (TLR9)in Chinese Han children from Zhejiang province, and their associations with asthma susceptibility and phenotypes. Methods A case-control study was conducted. A total of 312 asthmatic children aged between 1.9 and 11.6 and 339 age matched healthy controls were enrolled in this study from April 2007 to November 2008. The -1486 C/T in rs187084 and -1237 C/T in rs5743836 loci of the TLR9 gene were genotyped by direct DNA sequencing of the PCR products. Serum levels of IFN-γ, IL-12 and IL-4 were detected by enzyme linked immunosorbent assay. Serum levels of total IgE were detected by chemiluminescence, and serum levels of ildren (P＜0.01). The CC genotype had the lowest levels of serum IFN-γand the highest levels of serum IL-4 among the three genotypes. There were no significant differences in these cytokines among the healthy controls (P＞0.05). No statistical differences of serum IL-12 were found among the three genotypes in the two groups (P＞0.05). (4) There were no significant differences of total IgE (log-transformed) among the three genotypes in the asthmatic children (P＞0. 05). Conclusion The -1237 C/T polymorphism of TLR9 gene was not detected in Chinese Han children in this study. The - 1486 C/T polymorphism was associated with the levels of serum IFN-γ and IL-4 in children with asthma.However, there were no correlations between the -1486C/T polymorphism and serum IL-12 levels, total IgE levels or asthmatic susceptibility.     

Toll样受体9基因启动子单核苷酸多态性与儿童变应性哮喘的相关性

目的 研究Toll样受体9(toll-like receptor 9,TLR9)基因启动子区单核苷酸多态性在浙江汉族儿童中的分布,探讨其与哮喘易感性及其表型之间的相关性.方法 对312例变应性哮喘患儿(哮喘组)和339名健康儿童(对照组)采用DNA直接测序法检测TLR9基因-1486(rs187084)和-1237(rs5743836)单核苷酸多态性;采用ELISA法检测两组不同基因型血清干扰素γ(interferon-γ,IFN-γ)、白细胞介素12(interleukin-12,IL-12)和白细胞介素4(interleukin-4,IL-4)水平;采用化学发光法检测血清总免疫球蛋白E(immunoglobulin E,IgE)水平;采用酶免疫荧光法检测血清变应原特异IgE.结果 (1)哮喘组和对照组均存在-1486位点T→C突变,哮喘组TT、TC和CC 3种基因型的频率分别是3 8.8%、48.4%和12.8%,对照组分别是41.0%、44.3%和14.7%;未发现-1237位点存在多态性.(2)哮喘组和对照组-1486位点各基因型的频率分布差异无统计学意义(P＞0.05),年龄分层后比较差异也无统计学意义(P＞0.05).(3)哮喘组-1486位点3种基因型的血清IFN-γ和IL-4水平差异有统计学意义(P＜0.01),CC基因型的IFN-γ水平较低而IL-4水平较高;对照组2种细胞因子的差异无统计学意义(P＞0.05).哮喘组和对照组血清IL-12水平在3种基因型间差异均无统计学意义(P＞0.05).(4)哮喘组-1486位点不同基因型血清总IgE水平差异无统计学意义(P＞0.05).结论 浙江汉族儿童不存在TLR9基因-1237位点多态性.TLR9基因-1486 C/T位点单核苷酸多态性与浙江汉族儿童哮喘易感性、血清IL-12及总IgE水平无关;-1486 C/T位点多态性与哮喘患儿血清IFN-γ和IL-4水平有关联,CC基因型的IFN-γ水平较低而IL-4水平较高.

Abstract：

Objective To investigate the distribution characteristics of the single nucleotide polymorphisms (SNPs) in the promoter region of the toll-like receptor 9 gene (TLR9)in Chinese Han children from Zhejiang province, and their associations with asthma susceptibility and phenotypes. Methods A case-control study was conducted. A total of 312 asthmatic children aged between 1.9 and 11.6 and 339 age matched healthy controls were enrolled in this study from April 2007 to November 2008. The -1486 C/T in rs187084 and -1237 C/T in rs5743836 loci of the TLR9 gene were genotyped by direct DNA sequencing of the PCR products. Serum levels of IFN-γ, IL-12 and IL-4 were detected by enzyme linked immunosorbent assay. Serum levels of total IgE were detected by chemiluminescence, and serum levels of ildren (P＜0.01). The CC genotype had the lowest levels of serum IFN-γand the highest levels of serum IL-4 among the three genotypes. There were no significant differences in these cytokines among the healthy controls (P＞0.05). No statistical differences of serum IL-12 were found among the three genotypes in the two groups (P＞0.05). (4) There were no significant differences of total IgE (log-transformed) among the three genotypes in the asthmatic children (P＞0. 05). Conclusion The -1237 C/T polymorphism of TLR9 gene was not detected in Chinese Han children in this study. The - 1486 C/T polymorphism was associated with the levels of serum IFN-γ and IL-4 in children with asthma.However, there were no correlations between the -1486C/T polymorphism and serum IL-12 levels, total IgE levels or asthmatic susceptibility.     

Toll样受体9基因启动子单核苷酸多态性与儿童变应性哮喘的相关性

Objective To investigate the distribution characteristics of the single nucleotide polymorphisms (SNPs) in the promoter region of the toll-like receptor 9 gene (TLR9)in Chinese Han children from Zhejiang province, and their associations with asthma susceptibility and phenotypes. Methods A case-control study was conducted. A total of 312 asthmatic children aged between 1.9 and 11.6 and 339 age matched healthy controls were enrolled in this study from April 2007 to November 2008. The -1486 C/T in rs187084 and -1237 C/T in rs5743836 loci of the TLR9 gene were genotyped by direct DNA sequencing of the PCR products. Serum levels of IFN-γ, IL-12 and IL-4 were detected by enzyme linked immunosorbent assay. Serum levels of total IgE were detected by chemiluminescence, and serum levels of ildren (P＜0.01). The CC genotype had the lowest levels of serum IFN-γand the highest levels of serum IL-4 among the three genotypes. There were no significant differences in these cytokines among the healthy controls (P＞0.05). No statistical differences of serum IL-12 were found among the three genotypes in the two groups (P＞0.05). (4) There were no significant differences of total IgE (log-transformed) among the three genotypes in the asthmatic children (P＞0. 05). Conclusion The -1237 C/T polymorphism of TLR9 gene was not detected in Chinese Han children in this study. The - 1486 C/T polymorphism was associated with the levels of serum IFN-γ and IL-4 in children with asthma.However, there were no correlations between the -1486C/T polymorphism and serum IL-12 levels, total IgE levels or asthmatic susceptibility.     

纤维蛋白原Bβ基因启动区单核苷酸多态性及连锁不平衡分析

目的　探讨纤维蛋白原 B(fibrinogen,FGB)基因启动区 - 14 8C/ T、- 4 5 5 G/ A、- 85 4 G/ A3个位点单核苷酸多态性 (single nucleotide polymorphism,SNP)在中国南方汉族人群的分布特征及连锁不平衡关系。方法　应用聚合酶链反应 -限制性片段长度多态性技术结合 DNA测序分析检测 377名中国南方汉族人 FGBβ基因型和等位基因的分布频率 ,群体数理遗传学方法分析 FGBβ 3个基因位点 SNP的遗传平衡吻合度和相互间连锁不平衡关系。结果　 3个 FGBβ SNP位点等位基因频率分布符合 Hardy-Weinberg平衡。检出了 FGBβ3个位点 SNP的共 9种基因型 ,- 14 8CC、CT、TT基因型频率分别为0 .5 97、0 .35 8和 0 .0 4 5 ;- 4 5 5 G/ A SNP各基因型频率与 - 14 8C/ T SNP相同 ;- 85 4 GG、GA、AA基因型频率分别为 0 .82 0、0 .178和 0 .0 0 2。各 SNP位点的少见型等位基因频率分别是 0 .2 2 4 (- 14 8T)、0 .2 2 4 (-4 5 5 A)、0 .0 92 (- 85 4 A) ;常见型等位基因频率分别为 0 .776 (- 14 8C)、0 .776 (- 4 5 5 G)、0 .90 8(- 85 4 G)。男女性别间各基因型和等位基因分布频率差异无显著性 (P>0 .0 5 )。经连锁不平衡检验 ,- 14 8C与 - 4 5 5 GSNP为完全一致型 ,- 85 4 G/ A与 - 14 8C/ T、- 4 5 5 G/ A为随机分布。结     

Detection of three single nucleotide polymorphisms in the gene encoding mannose-binding lectin in a single pyrosequencing reaction

Mannose-binding lectin (MBL) is a key molecule of innate immunity. Binding of MBL to carbohydrates present on pathogens activates the lectin pathway of complement activation, resulting into opsonization and anti-microbial protection. Three frequently occurring single nucleotide polymorphisms (SNPs) are described in the coding region of the MBL2 gene that are associated with abnormal polymerization of the MBL molecule, decreased serum concentrations of high molecular weight MBL, and strongly impaired function. Clinical studies have shown that these MBL SNPs are associated with increased susceptibility to infections, especially in immune-compromised persons, as well as with accelerated progression of chronic diseases. The present study describes a novel method to detect the three major MBL SNPs by pyrosequencing. The close proximity of these SNPs allows their detection in one single pyrosequencing reaction, resulting in clearly distinguishable patterns for each allele combination described until now. This method can be used for the easy and reliable detection of MBL SNPs to identify the basis of functional MBL deficiency in clinical diagnostics and research.