非综合征型唇腭裂与MTHFR基因多态性的相关性研究

目的:研究亚甲基四氢叶酸还原酶(methylenetetrahydrofolate reductase,MTHFR)基因位点C677T和A1298C与中国江苏地区汉族人群非综合征型唇腭裂((nonsyndromic cleft lip with or without cleft palate,NSCL/P)发生的相关性...     

非综合征性唇腭裂与亚甲基四氢叶酸还原酶基因A1298C相关性的研究

目的 研究亚甲基四氢叶酸还原酶基因(methylenetetrahydrofolate reductase,MTHFR)A1298C多态性与中国华北人群非综合征性唇腭裂(non-syndromic cleft lip with or without cleft palate,NSCL/P)的关系.方法 通过聚合酶链反应-限制性片段长度多态性,在158例NSCL/P患者和192名健康对照中,对MTHFR基因A1298C单核苷酸多态性(single nucleotide polymorphism,SNP)rs1801131进行检测.利用拟合优度卡方检验分析基因型分布频率是否符合Hardy-Weinberg平衡定律;应用Unphased软件分析等位基因频率与NSCL/P的相关性.结果 MTHFR基因A1298C多态性基因型频率分布符合Hardy-Weinberg平衡;等位基因和基因型频率在唇裂合并或不合并腭裂组和健康对照组之间差异无统计学意义;基因型分布单纯腭裂(AA 78%、AC+CC 22%)与健康对照组(AA 74%、AC+CC 26%)比较,差异有统计学意义(χ2＝4.256,P=0.039),AC+CC基因型频率健康对照组(26%)高于单纯腭裂组(22%)(OR=0.8,95%CI=0.381～1.683).结论 MTHFR A1298C多态性位点可能与中国人群非综合征性单纯腭裂的发生有关.

Abstract：

Objective To investigate the association between a polymorphism of methylenetetrahydrofolate reductase with Non-syndromic cleft lip with or without cleft palate(NSCL/P)in Chinese population. Methods The polymerase chain reaction (PCR)-based restriction fragment length polymorphism(RFLP)technique was used to detect a single nucleotide polymorphism(SNP), rs1801131, at the methylenetetrahydrofolate reductase(MTHFR)gene in both 158 patients with NSCL/P and 192 healthy individuals. The Hardy-Weinberg equilibrium for genotypic distributions was estimated by the goodness-of-fit test. The UNPHASED program was applied to perform the association analysis. Results The genotypic distribution of A1298C was not deviated from the Hardy-Weinberg equilibrium in both controls and patients. No association was found between cleft lip with or without palate(CL/P)and controls. There was significant difference of cleft palate only(CPO)and the healthy individuals(χ2＝4.256, P=0.039). The frequency of AC+CC genotype was higher in control group than that in CPO group(OR=0.8, 95%CI=0.381-1.683),26 among 100 healthy individuals carried AC+CC genetypes,which were carried by 22% of CPO patients. Conclusions The polymorphism of MTHFR A1298C may be involved in the occurrence of non-syndromic cleft palate only in Chinese population.     

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目的    研究亚甲基四氢叶酸还原酶（MTHFR）基因 C677T多态性与山东地区非综合征性唇腭裂（NSCL／P）的相关性。方法    于2008 年 9月在山东省优生技术重点实验室采用聚合酶链反应-限制性片段长度多态性（ PCR-RFLP）分析,对2006 年8月至2008年8月曾在齐鲁医院治疗的来自山东地区NSCL／P患儿家庭34户和健康查体的正常儿童家庭46户的家庭成员MTHFR基因的C677T基因型进行检测。结果    携带T等位基因的父母,其子代患NSCL／P的危险性是不携带T等位基因父母的子代的2.420倍;母子都是TT突变纯合子,子代患NSCL／P的危险性是母子为非TT纯合子的4.162倍;子代是TT突变纯合子患NSCL／P的危险性是非TT纯合子的3.812倍。结论    山东地区NSCL／P与MTHFR基因 C677T的多态性相关,与父母的基因型存在联系;T基因在母子组合的研究中有统计学意义,父母传递给子代的T等位基因对后代的患病有重要作用。     

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目的探讨亚甲基四氢叶酸还原酶（MTHFR）基因A1298C多态性与山西人群非综合征性唇腭裂（nonsyndromic cleft lip with or without cleft palate,NSCL／P）的相关性。方法选取2010年9月至2012年4月山西地区150例NSCL／P患者及其父母作为病例组（其中有135个完整的NSCL／P核心家系）,150例正常新生儿作为对照组,应用聚合酶链式反应一限制性片段长度多态性（PCR—RFLP）分析方法,对MTHFR基因A1298C位点的多态性进行检测,利用人群关联研究分析、病例组核心家系的传递不平衡检验（TDT）、单体型的相对危险度（HHRR）分析来研究该突变与NSCL／P的相关性。结果病例组和对照组人群基因型均未偏离Hardy-Weinberg遗传平衡定律（P〉0．05）;病例组与对照组进行子代间比较,AA、AC、CC3种基因型分布差异有统计学意义（P〈0．05）,A等位基因和c突变等位基因的分布差异均有统计学意义（P〈0．05）;NSCL／P核心家系TDT检验,差异有统计学意义（P〈0．05）,表明突变等位基因c存在着传递失衡的现象;HHRR检验结果表明,MTHFR基因A1298C位点多态性由双亲传递给患病子女的等位基因（C／A）频率差异有统计学意义（P〈0．05）。结论MTHFR基因A1298C位点多态性与山西人群NSCL／P的发生存在关联。     

非综合征型唇腭裂与新疆维吾尔族、汉族 MTHFR 基因多态性的研究

目的：探讨新疆维吾尔族、汉族非综合征型唇腭裂与 MTHFR 基因多态性的关系和民族差异。方法：170例 NSCL／P儿童患者和100例健康儿童对照,SNaPshot 分型方法检测 rs1801131、rs1801133位点多态性,分析基因型、等位基因频率和2个位点联合作用与 NSCL／P 的关系以及民族间差异。结果：rs1801133TT 与 T 等位基因在维、汉两民族间有统计学差异（P ＜0．05）;rs1801133CT 与 CT ＋TT 基因型在总病例组和总对照组间均有统计学差异（P ＜0．05）;rs1801131和 rs1801133的联合分析在维族内、汉族内、两民族间、维汉总人群中病例组和对照组均有统计学差异（P ＜0．05）;rs1801131多态性在维族内、汉族内、两民族间以及维汉总人群中均无统计学差异（P ＞0．05）。结论：rs1801133TT 与 T 等位基因的汉族比维吾尔族更易罹患 NSCL／P;rs1801131AC 和 rs1801133CC 联合作用与 NSCL／P 相关且维吾尔族罹患风险高于汉族;rs1801131多态性与 NSCL／P 不相关并无民族差异;rs1801133CT 与 CT ＋TT 基因型均是避免疾病的保护因素。     

亚甲基四氢叶酸还原酶C677T多态性与非综合征性唇腭裂的关系

目的研究亚甲基四氢叶酸还原酶基因C677T多态性与非综合征性唇腭裂的关系。方法利用聚合酶链反应-限制性片段长度多态性方法（PCR-RFLP）,在168例非综合征性唇腭裂患者和192名正常对照中,对MTHFR基因C677T单核苷酸多态性（SNP,rs1801133）进行检测。利用拟合优度卡方检验,分析基因型分布频率是否符合Hardy-Weinberg平衡定律;应用UNPHASED软件包分析多态性位点与非综合征性唇腭裂的相关性。结果MTHFRC677T多态性基因型频率分布符合Hardy-Weinberg平衡;MTHFR C677T等位基因分布在NSCL／P组与对照组之间有显著性差异（P〈0.05）,正常组中T等位基因的频率明显高于NSCL／P组。结论MTHFR C677T多态性位点在中国人群中与非综合征性唇腭裂形成的发生相关联。     

染色体10q25位点多态性与中国非综合征性唇腭裂的关联研究

目的:研究染色体10q25上2个单核苷酸多态性(single nucleotide polymorphisms,SNP)位点rs7078160、rs4752028与中国人群非综合征性唇腭裂(non-syndromic cleft lip with or without cleft palate,NSCL/P)发病的相关性。方法:收集180例NSCL/P患者作为病例组,并按照表型分为单纯唇裂组、唇腭裂组、单纯腭裂组,将单纯唇裂组和单纯腭裂组合并为唇/腭裂组;选取360名健康人作为对照组。采集病例组和对照组的外周血血样并提取DNA。对上述2个SNP设计引物,PCR扩增其序列,通过二代测序进行基因型分型。利用SPSS19.0软件包中的χ2检验对病例组与对照组的基因型以及等位基因频率进行分析。结果:rs7078160的等位基因频率在唇/腭裂组与对照组中的差异最为显著(P=0.008,OR=1.500,95%CI=1.116~2.016),rs4752028位点的等位基因频率在唇/腭裂组和对照组间亦存在显著差异(P=0.028,OR=1.424,95%CI=1.041~1.948)。结论:染色体10q25区域的rs7078160和rs4752028位点与中国人群非综合征性唇腭裂的发病相关。     

非综合征性唇腭裂与MSX1基因传递不平衡研究

目的探讨MSX1基因与湖南汉族人群非综合征性唇腭裂（nonsyndromic cleft lip and palate，NSCLP）遗传易感性的关系。方法以MSX1基因内含子区的CA重复微卫星作为遗传标记，采用聚合酶链式反应（polymerase chain reaction，PCR）-变性聚丙烯酰胺凝胶（polyacrylamide gel electrophoresis，PAGE）基因分型技术对湖南汉族129个NSCLP核心家系387名成员进行基因型分析，并行传递不平衡检验（transmission disequilibrium test，TDT）及Logistic回归分析。结果TDT分析显示，MSX1基因CA4等位基因在唇裂伴（不伴）腭裂（cleft lip with or without palate，CL／P）和单纯性腭裂（cleft palate only，CPO）组均被优势传递给患病后代（P=0．018，P=0．041）。Logistic回归分析结果支持隐性遗传模式，CA4本身或其作为一致病基因的遗传标志以隐性遗传模式被遗传（P=0．009）。结论MSX1基因与湖南汉族人群NSCLP相关联，可能是其易感基因或与之存在连锁不平衡。     

侧腭突生长发育和腭裂的分子调节机制

唇腭裂是一类常见的先天性畸形．可单独发生,也可与300多种已知的畸形伴发于综合征。唇腭裂又分为4型：综合征性唇裂伴或不伴腭裂（cleft of lip with or without palate,CL／P）、综合征性腭裂（cleft palat,CPO）、非综合征性唇裂伴或不伴腭裂（nonsyndromic cleft of lip with or without palate．nsCL／P）和非综合征性腭裂（nonsyndromic cleft palate．nsCPO）．     

WNT10A基因rs10177996位点单核苷酸多态性在新疆地区维族和汉族人群中的分布

目的:研究新疆地区维吾尔族和汉族人群中WNT10A基因rs10177996位点单核苷酸多态性的分布特征。方法:采用横断面调查,以新疆乌鲁木齐地区154例汉族个体及新疆喀什地区134例维吾尔族个体为研究对象,采用棉拭子法获取颊黏膜脱落的上皮细胞,用专用试剂盒提取DNA并检测,利用PCR（酶合聚链反应）技术扩增合格样品中含有的相应SNP的序列片段,基因测序后,结合软件进行基因分型。采用SPSS 23.0软件包对所得数据进行统计学分析。结果:新疆喀什地区维吾尔族人群中WNT10A基因rs10177996位点CC、CT和TT的基因型频率分别为8.21%、30.60%和61.19% ,C/T等位基因频率分别为C=23.51%,T=76.49%。新疆乌鲁木齐地区汉族人群中WNT10A基因rs10177996位点CC、CT和TT的基因型频率分别为9.74%、43.51%和46.75%,C/T等位基因频率分别为C=31.49%,T=68.51%。新疆喀什地区维吾尔族人群与新疆乌鲁木齐地区汉族人群相比,TT型基因型频率分布高,CC型分布低,差异具有显著性（P=0.046）。新疆喀什地区维吾尔族人群与欧洲人群相比,维吾尔族人群中SNP位点的TT型基因型频率分布低,CC型分布高,差异具有显著性（P=0.050）。新疆乌鲁木齐地区汉族人群与欧洲人群相比,TT型基因型频率分布低,CC型分布高,差异具有显著性（P<0.01）。在上述3种人群中,关于C/T等位基因频率的分布,新疆乌鲁木齐地区汉族人群与欧洲人群相比,C等位基因频率的分布高,T等位基因频率的分布低,差异具有显著性（P=0.033）。但新疆喀什地区维吾尔族人群与新疆乌鲁木齐地区汉族人群比较,以及新疆喀什地区维吾尔族人群与欧洲人群比较,均无显著差异（P>0.05）。新疆乌鲁木齐地区汉族人群和新疆喀什地区维吾尔族人群不同性别间也无显著差异（P>0.05）。结论:在新疆乌鲁木齐地区汉族人群、新疆喀什地区维吾尔族人群以及已报道的欧洲人群中,WNT10A基因rs10177996位点单核苷酸多态性分布特征各不相同。     

Association between methylenetetrahydrofolate reductase polymorphisms and non-syndromic cleft lip with or without palate susceptibility: an updated systematic review and meta-analysis

Methylenetetrahydrofolate reductase (MTHFR) polymorphisms are thought to be involved in the development of cleft lip with or without cleft palate (NSCL/P), but published results are contradictory. We therefore designed an updated meta-analysis to pool eligible studies and to evaluate further the possible relations between MTHFR polymorphisms (c.677C > T and c.1298A > C) and susceptibility to NSCL/P. A comprehensive search based on PubMed, Medline, Web of Science, and Embase databases was made up to February 2018. Twenty-three case-control and 10 case-parent trio studies (including 1149 cases and 1161 controls) were retrieved. Odds ratio (OR) with 95% CI were used to estimate the pooled strength of association under different genetic models. The Q test and I² test were used to estimate heterogeneity among studies, the quality of which was assessed using the Newcastle-Ottawa scale. In the MTHFR c.677C > T polymorphism group, there were significant overall results for the recessive (OR 1.231, 95%CI 1.092 to 1.387) and homozygote (OR 1.252, 95%CI 1.078 to 1.456) models. Subgroup analysis by subjects and ethnicity identified only associations in European mothers for the recessive model and the homozygote model. For the c.1298A > C group, there were no significant results for either European or Asian patients for all genetic models. The MTHFR c.677C > T polymorphism might increase susceptibility to NSCL/P in European mothers, but was negatively associated in Asian patients, and the MTHFR c.1298A > C polymorphism is not involved in the development of NSCL/P in either European or Asian patients.     

5,10‐Methylenetetrahydrofolate reductase single nucleotide polymorphisms and gene–environment interaction analysis in non‐syndromic cleft lip/palate

Non‐syndromic cleft lip/palate (NSCL/P) is a common congenital defect in Mexico. Periconceptional intake of folic acid (FA) may reduce the risk of this malformation. Although the 5,10‐methylenetetrahydrofolate reductase (MTHFR) enzyme participates in folate metabolism, several studies failed to find any association between NSCL/P and the MTHFR C677T and A1298C polymorphisms. However, interactions among NSCL/P, MTHFR gene polymorphisms, and FA intake have not been explored in Mexican populations. This case–control study included 132 patients with NSCL/P and 370 controls from Mexico City. Maternal FA consumption during pregnancy was examined, as were the MTHFR C677T and A1298C polymorphisms and gene–FA interactions. Maternal FA intake during the periconceptional period was lower in cases (1.5%) than in controls (13%), with the risk of delivering a child with NSCL/P lower in mothers who consumed FA (OR = 0.29, 95% CI: 0.19–0.44). In addition, the risk of NSCL/P was lower in children with the TT than the CC genotype of MTHFR C677T (OR = 0.39, 95% CI: 0.23–0.68), after Bonferroni correction and exclusion of stratification. No evidence of gene–FA interaction was found. These results indicate that maternal FA intake and the TT genotype of the MTHFR C677T polymorphism in children independently reduced the risk of NSCL/P in our population.     

山西人群MTHFR基因rs1801133位点多态性与非综合征性唇腭裂的关联性研究

目的:研究亚甲基四氢叶酸还原酶(methylenetetrahydrofolate reductase,MTHFR)基因rs1801133位点多态性与非综合征性唇腭裂(NSCL/P)的关系。方法:采用聚合酶链反应-限制性片段长度多态性方法,检测334例非综合征性唇腭裂患者和314例正常对照组的MTHFR基因rs1801133位点的多态性。结果:MTHFR基因rs1801133位点基因型频率分布符合Hardy-Weinberg平衡;MTHFR基因rs1801133位点基因型及等位基因的分布在NSCL/P组与对照组之间均无统计学意义(P>0.05)。结论:MTHFR基因rs1801133位点多态性与山西人群非综合征性唇腭裂的发生无关。     

VAX1 gene associated non-syndromic cleft lip with or without palate in Western Han Chinese

ObjectiveNon-syndromic cleft lip with or without palate (NSCL/P) is one of the most common human birth defects, it results from multiple genetic and environmental risk factors. Recently, GWA studies identified associations between NSCL/P and two genetic risk loci, rs7078160 and rs4752028, at VAX1.DesignCurrently, we tried to investigate the roles of the two loci among 302 NSCL/P trios (129 non-syndromic cleft lip only (NSCLO) trios and 173 non-syndromic cleft lip and cleft palate (NSCLP) trios) from Western Han Chinese. The two SNPs were genotyped by SNPscan method; Hardy–Weinberg equilibrium test, allelic TDT and parent-of-origin effect were performed by PLINK software, and genotypic TDT and haplotype by FBAT software.ResultsAllelic TDT analysis revealed allele A at rs7078160 was over-transmitted among NSCL/P group (P = 0.0086, OR_transmission = 1.36, 95%CI: 1.08–1.72). Parent-of-origin effect analysis revealed a paternal special over-transmission of allele A at rs708260 in NSCL/P group (P = 0.0079). Haplotype AC of rs7078160-rs4752028 was significant over-transmitted in the NSCL/P group.ConclusionsOur study firstly confirmed that allele A at rs7078160 at VAX1 gene was a risk factor for NSCL/P in Western Han Chinese population.     

中国人群非综合征性唇腭裂患者IRF6基因突变检测

目的 探讨干扰素调节因子6(interferon regulatory factor 6, IRF6) 在非综合征性唇腭裂(non-sydromic cleft lip and/or cleft palate,NSCL/P)患者中的突变情况。方法：收集119例NSCL/P患者及288名健康人对照样本的外周血血样并提取DNA。在IRF6基因的全部外显子分别设计引物,PCR扩增其序列,通过测序找出IRF6基因突变,并将这些突变在对照样本中进行验证。结果：共发现5种在正常人中没有的突变,其中4种是新发现的突变。结论：IRF6基因突变在中国人群中参与了非综合征唇腭裂疾病的发生。     

宁夏回汉族人群Wnt3基因单核苷酸多态性与非综合征型唇腭裂相关性的研究

目的 探讨宁夏回汉族人群中Wnt3基因rs142167和rs7216231位点单核苷酸多态性（SNP）与非综合征型唇腭裂（NSCL/P）的相关性。方法 收集宁夏地区回汉族人群非综合征型唇腭裂患者371例为病例组,其中汉族患者166例,回族患者205例;收集患者父亲196例,患者母亲224例,其中150例患者为NSCL/P核心家系;258例健康新生儿为对照组,其中汉族190例,回族68例。采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法检测Wnt3基因多态位点rs142167和rs7216231基因型,对比分析2组的基因型和等位基因,并进行传递不平衡检验（TDT）和以家系为基础的相关性检验（FBAT）分析。结果 回汉族人群病例组与对照组比较及其民族分层比较,唇裂、腭裂、唇腭裂及总病例组rs142167和rs7216231位点均无统计学差异（P＞0.05）。TDT分析结果显示：rs142167和rs7216231位点的等位基因均不存在过传递（P＞0.05）。FBAT分析结果显示：单倍型G-G具有统计学意义（P<0.05）。结论 Wnt3基因多态性与宁夏地区回汉族人群非综合征型唇腭裂不存在相关性。