RF6基因多态性与黔北人群非综合征性唇腭裂的相关性研究

目的 探讨黔北地区人群IRF6基因rs2235371和rs2235373 SNP位点的多态性及其与非综合征性唇腭裂的相关性。方法 采用PCR和测序方法对153个对照儿童和123个NSCL/P儿童的IRF6基因中2个SNP位点rs2235371和rs2235373进行扩增和测序;对样本群体进行Hardy-Weinberg平衡分析,比较2组人群的基因型频率、等位基因频率及OR分析;两位点连锁不平衡分析。结果 对照组与病例组人群rs2235371基因型均含有GG、GA和AA型,rs2235373位点均含有CC、CT和TT型。对于2个位点,对照组和病例组均符合Hardy-Weinberg平衡法则(P＞0.05)。2组人群中,rs2235371和rs2235373位点的等位基因和基因型差异均有统计学意义(P＜0.05);rs2235371位点GGvsGA的OR值(95%CI) =1.725(1.025～2.902),GGvsAA的OR值(95%CI) = 2.100(1.109～4.328);rs2235373位点CCvsTT的OR值(95%CI) = 2.263(1.348～5.015),CTvsTT的OR值(95%CI) = 2.061(1.108～2.169);(P＜0.05)。rs2235371和rs2235373位点存在连锁不平衡,GC单倍型是主要的单倍体型,对NSCL/P均有致病风险,OR值(95%CI)= 1.722(1.219～2.431), (P＜0.05)。结论 在黔北地区人群中,IRF6基因 rs2235371和rs2235373位点均具有多态性;rs2235371位点的GG基因型和rs2235373位点的CC、CT基因型与NSCL/P的发生有相关性;rs2235371和rs2235373位点存在连锁不平衡,GC单倍型对NSCL/P有致病风险。     

新疆维、汉两民族非综合征性唇腭裂IRF6基因多态性分析

目的:探讨非综合征性唇腭裂(NSCL/P)干扰素调节因子6(IRF6)rs2013162位点单核苷酸多态性(SNP)在新疆维、汉两民族内和民族间基因型和等位基因型的频率差异。方法:抽取100例NSCL/P患者作为NSCL/P组(维吾尔族50例、汉族50例),对照组100例(维吾尔族50例、汉族50例),运用聚合酶链式反应-限制性片段长度多态性(PCR-RELF)技术来分析IRF6基因的多态性,病例-对照研究分析两组基因型和等位基因型频率及两民族内和民族间频率的差异。结果:rs2013162位点GG基因型和等位基因G和T频率在NSCL/P组和对照组中分布差异有统计学意义(P<0.05),维、汉两民族内rs2013162位点维吾尔族中GG和TT基因型及等位基因G和T分布差异有统计学意义(P<0.05),维、汉两民族间rs2013162位点NSCL/P组中维吾尔族GG和TT基因型和等位基因G的频率均高于汉族,两民族间基因型和等位基因型分布差异均无统计学意义(P>0.05)。结论:新疆维、汉族NSCL/P与rs2013162位点GG基因型及等位基因G存在相关性。     

还原叶酸载体1基因A80G多态性与非综合征型唇腭裂的相关性研究

目的探讨还原叶酸载体(RFC)1基因A80G多态性与非综合症型唇腭裂(NSCL/P)相关性。方法收集97个核心家庭和104个对照家庭,用聚合酶链式反应-限制性片段长度多态性方法,进行RFC1基因A80G位点多态性检测,用人群关联研究分析、NSCL/P核心家庭的TDT、HHRR、FBAT等检验统计分析。结果人群关联研究分析,子代、父亲、母亲病例组和对照组之间基因型和等位基因的分布差异无显著性(P>0.05)。AG基因型相对于AA基因型的比值比OR(95%CI)、P值分别为子代0.87(0.44～1.70)、0.657;父亲1.09(0.54～2.21)、0.788;母亲1.63(0.79～3.36)、0.152。GG基因型相对于AA基因型的OR(95%CI)、P值分别为子代0.48(0.19～1.23)、0.094;父亲0.93(0.38～2.23)、0.850;母亲1.30(0.46～3.67)、0.584。G基因相对于A基因的OR(95%CI)、P值分别为子代1.22(0.78～1.94)、0.386;父亲1.02(0.64～1.61)、0.945;母亲0.91(0.58～1.41)、0.660。携带有突变基因G并不能增加患NSCL/P的危险。NSCL/P核心家庭分析,TDT检验中传递G等位基因给患病子代的为40次,传递A等位基因的为71次,等位基因A比突变等位基因G更易传递给患病子代(χ2=8.658,P<0.05;HHRR检验χ2=10.31,P<0.05;FBAT检验Z=2.942,P<0.05)。结论利用核心家庭资料进行统计分析的结果则认为RFC1基因A80G位点变异存在传递不平衡现象,这与NSCL/P发病危险之间存在有一定的关联关系,等位基因A可能与NSCL/P的高危显性有关系。     

TGFA基因多态性与NSCL/P的遗传易感性

目的:探讨中国汉族人TGFA基因的多态性对非综合征性唇腭裂的遗传易感性的作用。方法:76例核心家庭和60例正常对照儿童的TGFA基因型,采用病例对照研究和传递不平衡检验进行分析。结果:病例对照研究结果表示:(2χ=7.77,P<0.05),TDT(2χ=5.26,P<0.05))。结论:TGFA C2等位基因在NSCL/P中存在连锁不平衡,TGFA基因多态性可能是中国汉族人NSCL/P的遗传易感性因素。     

Evidence for SNP‐SNP interaction identified through targeted sequencing of cleft case‐parent trios

Nonsyndromic cleft lip with or without cleft palate (NSCL/P) is the most common craniofacial birth defect in humans, affecting 1 in 700 live births. This malformation has a complex etiology where multiple genes and several environmental factors influence risk. At least a dozen different genes have been confirmed to be associated with risk of NSCL/P in previous studies. However, all the known genetic risk factors cannot fully explain the observed heritability of NSCL/P, and several authors have suggested gene‐gene (G × G) interaction may be important in the etiology of this complex and heterogeneous malformation. We tested for G × G interactions using common single nucleotide polymorphic (SNP) markers from targeted sequencing in 13 regions identified by previous studies spanning 6.3 Mb of the genome in a study of 1,498 NSCL/P case‐parent trios. We used the R‐package trio to assess interactions between polymorphic markers in different genes, using a 1 degree of freedom (1df) test for screening, and a 4 degree of freedom (4df) test to assess statistical significance of epistatic interactions. To adjust for multiple comparisons, we performed permutation tests. The most significant interaction was observed between rs6029315 in MAFB and rs6681355 in IRF6 (4df P = 3.8 × 10^?8) in case‐parent trios of European ancestry, which remained significant after correcting for multiple comparisons. However, no significant interaction was detected in trios of Asian ancestry.     

六号染色体短臂MHC区DRB3、DRB1基因多态性与精神分裂症症状的关系探讨

目的 探讨六号染色体短臂MHC区DRB3、DRBl基因多态性与精神分裂症症状的相关性。方法 采用聚合酶链反应(PCR)和限制性内切酶片段长度多态性(RFLP)方法检测两个基因位点上的单核苷酸多态性(SNPs)，并对116例精神分裂症患者家系进行连锁不平衡分析。结果 DRBl的SNP(rs707954：G／T碱基互换)等位基因与关系妄想症状呈显著相关(X^2=5．484，df=l，P=0．019)，GG、GT、TT三种基因型频率在关系妄想症状中呈显著相关(X^2=6．771，df=2，P=0．034)。rs707954的三种基因型频率与情感淡漠症状呈显著相关(X^2=12．110，df=4，P=0．017)。结论 DRBl位点等位基因与关系妄想和情感淡漠症状高度相关，DRBl基因型与关系妄想症状高度相关。     

中国人群WNT代谢通路与非综合征型唇腭裂的亲源效应分析

目的 非综合征型唇裂合并或不合并腭裂（NSCL/P）是一类常见的出生缺陷,遗传致病因素一直是其病因学研究的热点。本研究拟基于家系设计在WNT代谢通路基因中探索亲源效应对NSCL/P发病风险的影响。方法 本研究人群为“唇腭裂的基因组学国际合作组研究”项目在中国地区募集的806个NSCL/P核心家系。利用对数线性模型探索WNT基因及其单体型的亲源效应与疾病的关联,采用Wald检验探索亲源效应与环境因素的交互作用。经过Bonferroni多重检验校正后,统计学检验的显著性阈值设为P<3.47×10^-4。结果 质量控制后共纳入7个基因上144个单核苷酸多态性位点进行分析。结果显示,NSCL/P家系中有8个位点具有潜在的亲源效应（P<0.05）,但经Bonferroni多重检验校正后,均未达到统计学显著性水平（P>3.47×10^-4）。NSCL/P家系中位于WNT9A rs4074668-rs12725747单体型（T-A）具有亲源效应,且经Bonferroni校正后差异仍有统计学意义（P=2.74×10^-4）。但该单体型的亲源效应与环境因素（被动吸烟、复合维生素补充）的交互作用并未达到统计学显著水平。结论 WNT代谢通路基因可能通过亲源效应影响NSCL/P的发生风险。位于WNT9A基因rs4074668-rs12725747单体型（T-A）亲源效应与NSCL/P发病风险存在显著关联。未来仍需其他独立样本验证以进一步确认WNT代谢通路在NSCL/P发生中的作用。     

非综合征性唇腭裂部分基因SNPs研究进展

非综合征性唇裂伴或不伴腭裂是人类最常见的先天性畸形之一,是一种遗传、环境因素及两者相互作用所致的多基因多因素遗传疾病.单核苷酸多态性是新一代遗传标记,可被用来寻找各种致病基因,目前认为单核苷酸多态性及其特定组合可能是造成以多基因多因素遗传病为代表的复杂性状疾病易感性的重要原因.     

转化生长因子α基因多态性与唇腭裂关联的研究

目的 探讨中国部分地区人群非综合征型唇裂伴或不伴腭裂（nsCL/P）与转化生长因子α基因（TGFα）TaqI位点多态性之间的关系。方法 采用聚合酶链反应限制片段长度多态性分析方法，对149个nsCL／P核心家庭成员DNA标本进行TGFα Taq I突变位点的基因型检测。利用传递失衡检验和以家庭为基础的关联研究（FBAT）方法，分析TGFα Taq I突变与nsCL／P发生之间的关系。结果 未发现TGFα Taq I突变的致病晓等位基因在nsCL／P核心家庭成员中存在传递不平衡（P〉0．05）；采用FBAT分析，未发现C2等位基因及C2C1基因型与nsCL／P发病危险之间关联有统计学意义（P〉0．05）。结论 TGFα Taq I突变可能不是中国部分地区人群nsCL／P发生的易感基因。     

Genetic variants in IRF6 and the risk of facial clefts: single-marker and haplotype-based analyses in a population-based case-control study of facial clefts in Norway

Mutations in the gene encoding interferon regulatory factor 6 (IRF6) underlie a common form of syndromic clefting known as Van der Woude syndrome. Lip pits and missing teeth are the only additional features distinguishing the syndrome from isolated clefts. Van der Woude syndrome, therefore, provides an excellent model for studying the isolated forms of clefting. From a population-based case-control study of facial clefts in Norway (1996-2001), we selected 377 cleft lip with or without cleft palate (CL/P), 196 cleft palate only (CPO), and 763 control infant-parent triads for analysis. We genotyped six single nucleotide polymorphisms within the IRF6 locus and estimated the relative risks (RR) conferred on the child by alleles and haplotypes of the child and of the mother. On the whole, there were strong statistical associations with CL/P but not CPO in our data. In single-marker analyses, mothers with a double-dose of the 'a'-allele at rs4844880 had an increased risk of having a child with CL/P (RR=1.85, 95% confidence interval: 1.04-3.25; P=0.036). An RR of 0.38 (95% confidence interval: 0.16-0.92; P=0.031) was obtained when the child carried a single-dose of the 'a'-allele at rs2235371 (the p.V274I polymorphism). The P-value for the overall test was <0.001. In haplotype analyses, several of the fetal and maternal haplotype relative risks were statistically significant individually but were not strong enough to show up on the overall test (P=0.113). Taken together, these findings further support a role for IRF6 variants in clefting of the lip and provide specific risk estimates in a Norwegian population.     

Identification of 16q21 as a modifier of nonsyndromic orofacial cleft phenotypes

Orofacial clefts (OFCs) are common, complex birth defects with extremely heterogeneous phenotypic presentations. Two common subtypes—cleft lip alone (CL) and CL plus cleft palate (CLP)—are typically grouped into a single phenotype for genetic analysis (i.e., CL with or without cleft palate, CL/P). However, mounting evidence suggests there may be unique underlying pathophysiology and/or genetic modifiers influencing expression of these two phenotypes. To this end, we performed a genome‐wide scan for genetic modifiers by directly comparing 450 CL cases with 1,692 CLP cases from 18 recruitment sites across 13 countries from North America, Central or South America, Asia, Europe, and Africa. We identified a region on 16q21 that is strongly associated with different cleft type (P  = 5.611 × 10^?8). We also identified significant evidence of gene–gene interactions between this modifier locus and two recognized CL/P risk loci: 8q21 and 9q22 (FOXE1 ) (P  = 0.012 and 0.023, respectively). Single nucleotide polymorphism (SNPs) in the 16q21 modifier locus demonstrated significant association with CL over CLP. The marker alleles on 16q21 that increased risk for CL were found at highest frequencies among individuals with a family history of CL (P  = 0.003). Our results demonstrate the existence of modifiers for which type of OFC develops and suggest plausible elements responsible for phenotypic heterogeneity, further elucidating the complex genetic architecture of OFCs.     

乙型肝炎患者人类白细胞抗原-DRB1、-DQA1、-DQB1等位基因多态性分析

目的 探讨人类白细胞抗原(HLA)-DRB1、-DQA1和-DQB1等位基因多态性与乙型肝炎(乙肝)之间的关系。方法 采用聚合酶链反应/序列特异性引物(PCR/SSP)技术对52例慢性乙肝患者、30例急性乙肝患者和106名正常人的HLA-DRB1、-DQA1和DQB1等位基因多态性进行了分析。结果 HLA-DRB1*0301、-DQA1*0501和-DQB1*0301在慢性乙肝患者组的等位基因频率(17.31％、25.96％、35.58％)明显高于正常对照组(5.67％、13.36％、18.87％),两者相比差异有显著性(X_1~2=12.3068,P_(c1)=0.0074;X_2~2=9.2002,P_(c2)=0.0157;X_3~2=15.5938,P_(c3)=0.0075)。HLA-DRB1*1101/1104和-DQA1*0301在慢性乙肝患者组的等位基因频率(0.96％、14.42％)明显低于急性乙肝患者组(13.33％、30％),两者相比差异有显著性(X_1~2=11.9206,P_(c1)=0.0145;X_2~2=8.7396,P_(c2)=0.0167)。结论 HLA-DRB1*0301、-DQA1*0501和-DQB1*0301与慢性乙肝患者的易感性密切相关,HLA-DRB1*1101/1104和-DQA1*0301与慢性乙肝的抗性密切相关,说明宿主的HLA-Ⅱ类基因是决定乙肝病毒感染转归的重要因素。     

转化生长因子α基因多态性和父亲吸烟的交互作用与唇腭裂发生的关联研究

目的探讨中国部分地区人群非综合征型唇裂伴或不伴腭裂（nsCL／P）与转化生长因子α基因（TGFα）TaqI位点多态性之间的关系,及其与父亲吸烟之间的交互作用。方法采用PCR-RFLP方法,对170个nsCL／P核心家庭成员DNA标本进行TGFα TaqI突变位点的基因型检测。利用TDT检验分析该突变与nsCL／P发生之间的关系,采用TDT检验的logistic回归模型分析TGFα基因突变与父亲吸烟之间的交互作用。结果未发现TGFα TaqI突变的致病C2等位基因在nsCL／P核心家庭成员中存在传递不平衡,但是父亲吸烟的nsCL／P核心家庭中C2C1基因型的父母将致病的C2等位基因传递给子代的频率是父亲不吸烟的nsCL／P核心家庭父母的约1／5（0．062～0．711）,控制其他环境因素后发现,父亲是否吸烟与TGFα TaqI突变位点C2等位基因传递之间是偏离乘法模型的负交互作用OR=0．102（0．017～0．619）。结论父亲是否吸烟与中国部分地区人群TGFα基因突变存在交互作用,但还有待于进一步研究加以验证。     

非综合征型单纯腭裂遗传流行病学研究进展

单纯腭裂是一种较为常见的出生缺陷,其中非综合征型单纯腭裂（NSCPO）占50%。NSCPO是受遗传和环境共同作用的复杂疾病,与非综合征型唇裂伴或不伴腭裂（NSCL/P）不同,通过全基因组关联研究发现的与NSCPO相关的常见遗传变异相对较少。本文对NSCPO的遗传流行病学研究进展进行综述。根据现有研究证据将已发现的NSC...     

胞浆型磷脂酶A2家族基因多态性与精神分裂症的关联性研究

目的 探讨中国北方汉族人群胞浆型磷脂酶A2(cPLA2)家族基因多态性与精神分裂症的遗传关联性.方法 采用聚合酶链反应(PCR)和连接酶检测反应(LDR)方法,在201个精神分裂症患者核心家系中检测cPLA2家族基因上的10个单核苷酸多态性(SNPs),对结果进行单倍型相对风险分析(HRR)、传递不平衡分析(TDT)、单倍型分析和多位点联合分析.结果 各位点在精神分裂症病例组和对照组中基因型分布均符合Hardy-Weinberg平衡.HRR和TDT分析表明,检测的10个SNPs位点与精神分裂症无关联性(P＞0.05).单倍型分析结果显示,由同一染色体上各位点组成的单倍型与精神分裂症均无关联性(P＞0.05).多位点联合作用分析显示,rs2162886与rsl668589,rs891014与rsl668589,rs2307279与rs7542180位点的联合作用与精神分裂症相关联(χ²=6.913,P=0.032;χ²=8.393,P=0.015;χ²=8.447,P=0.038).结论 cPLA2家族基因中存在多个与精神分裂症关联的易感位点.     

候选基因SNP位点与非综合征性唇腭裂的相关性

目的探讨亲代12个候选基因SNP位点与子代非综合征性唇腭裂(NSCLP)的关系,为开展NSCLP的基因诊断和针对性的防治提供基础资料。方法选取41例NSCLP患儿母亲和109例非患儿母亲分别作为病例组、对照组,提取其血液中DNA,采用改良多重高温连接酶检测法(iMLDR)对CENPJ、c14orf49和YOD1等12个基因的SNP位点进行基因分型,计算各基因型在病例组、对照组中的频率分布,并采用χ2检验分析12个SNP位点基因型与NSCLP的遗传关系。结果 C14orf49、EIF2B3、HEATR8、KIF20B、PARVA、PKP1、RECQL5、REG3A、SEC16A、TEX11基因的10个SNP位点的基因型频率分布在病例组、对照组间的差异均无统计学意义(P值均0.05)。CENPJ基因rs35498994位点的基因型C在病例组、对照组中分布频率分别为46%、33%;YOD1基因chr1_207224322位点的基因型GT和T在病例组分布频率分别为7%、4%,在对照组中分布频率均为0%,以上差异均有统计学意义(P值均0.05)。结论母亲携带CENPJ基因(rs35498994)等位基因C,或YOD1(chr1_207224322)突变基因型GT和T会增加子代患NSCLP的风险。     

矽肺与人类白细胞抗原-DRB1·和-DQB1·基因相关性的研究

目的 探讨我国北方汉族人矽肺的易感性与人类白细胞抗原（HLA）－DRB1＊、DQB1＊位点等位基因多态的相关性。方法 用序列特异性引物的聚合酶链式反应（PCR－SSP）方法，分析48名汉族矽肺患者及100名汉族无血缘关系对照者（均为有过14年以上接尘史的井下凿岩工）HLA－DRB1＊、DQB1＊位点的基因频率分布，以相对危险度（RR）代表相关程度。结果 在矽肺患者组中，DRB1＊1401、DQB1＊05的等位基因频率明显高于对照组，经统计学处理，两组间差异具有显著性（χ^2=5.61,P=0.0066,RR=17.40;χ^2=10.70,P=0.0011,RR=3.81)；而DRB1＊09的等位基因频率则明显低于对照组，差异有显著性（χ^2=5.70,P=0.0187,RR=0.21)；其他等位基因频率的分布差异无显著性（P＞0．05）。结论 HLA－DRB1＊1401、DQB1＊05可能与矽肺的易感性有关，而DRB1＊09则可能与机体抗矽肺的保护性有关。HLA－DR位点与机体易感及保护的双重作用有关，等位基因之间的共同作用可能是影响矽肺发生的原因之一。     

Examining markers in 8q24 to explain differences in evidence for association with cleft lip with/without cleft palate between Asians and Europeans

In a recent genome-wide association study (GWAS) from an international consortium, evidence of linkage and association in chr8q24 was much stronger among nonsyndromic cleft lip/palate (CL/P) case-parent trios of European ancestry than among trios of Asian ancestry. We examined marker information content and haplotype diversity across 13 recruitment sites (from Europe, United States, and Asia) separately, and conducted principal components analysis (PCA) on parents. As expected, PCA revealed large genetic distances between Europeans and Asians, and a north-south cline from Korea to Singapore in Asia, with Filipino parents forming a somewhat distinct Southeast Asian cluster. Hierarchical clustering of SNP heterozygosity revealed two major clades consistent with PCA results. All genotyped SNPs giving P < 10(-6) in the allelic transmission disequilibrium test (TDT) showed higher heterozygosity in Europeans than Asians. On average, European ancestry parents had higher haplotype diversity than Asians. Imputing additional variants across chr8q24 increased the strength of statistical evidence among Europeans and also revealed a significant signal among Asians (although it did not reach genome-wide significance). Tests for SNP-population interaction were negative, indicating the lack of strong signal for 8q24 in families of Asian ancestry was not due to any distinct genetic effect, but could simply reflect low power due to lower allele frequencies in Asians.     

Maternal reproductive history and the risk of isolated congenital malformations

We examined the relationship between maternal reproductive history and the newborn's risk of isolated congenital malformations in a large case-control cohort from the Polish Registry of Congenital Malformations. Congenital malformations were classified into four categories: isolated congenital heart defects (n=1673), isolated cleft palate (n=255), cleft lip with or without cleft palate (n=448) and renal agenesis (n=103). The case groups were compared with a shared group of 2068 controls recruited in the same time period and geographic area. Multivariable logistic regression was used to assess the risk associated with maternal gravidity and of previous miscarriages after accounting for maternal age and other potential risk factors. In unadjusted analyses, maternal gravidity was significantly associated with increased risk of all four classes of congenital malformations. After adjustment, a significant association persisted for congenital heart defects [odds ratio (OR)=1.22, [95% confidence interval (CI) 1.09, 1.36], P=0.0007] and cleft lip with or without cleft palate (OR=1.21, [95% CI 1.09, 1.36], P=0.0005). A similar trend existed for isolated cleft palate (OR=1.18, [95% CI 1.02, 1.37], P=0.03). There was no appreciable increase in the risk of congenital malformations associated with a maternal history of miscarriages, but a trend for a protective effect on the occurrence of cleft lip with or without cleft palate was observed (OR=0.72, [95% CI 0.52, 0.99], P=0.045). Based on our data, maternal gravidity represents a significant risk factor for congenital heart defects and cleft lip with or without cleft palate in the newborn infant. Our data do not support an increase in risk because of past history of miscarriages.