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）．     

宁夏人群MSX1基因CA重复序列多态性与非综合征型唇腭裂相关性的研究

目的 探讨宁夏人群同源异型盒1(MSX1)基因CA重复序列(STR)与非综合征型唇腭裂的相关性.方法 收集宁夏地区非综合征型唇腭裂三人核心家庭(患儿及其双亲)40例,采用聚合酶链反应-单链构象多态性(PCR-SSCP)方法 检测MSX1基因CA 重复序列基因型,进行传递不平衡检验(TDT)和家系为基础的相关性分析(FBAT).结果 运用TDT发现,MSX1基因CA重复序列CA4等位基因在本研究人群非综合征型唇腭裂患儿中存在过传递(P=0.034).FBAT分析表明MSX1基因CA重复序列与本研究人群非综合征型唇腭裂具有相关性(P＜0.05).结论 MSX1基因CA重复序列与宁夏人群非综合征型唇腭裂存在相关性.     

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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。     

先天性唇腭裂发病危险因素的Logistic回归分析

目的 探讨先天性唇腭裂（cleft lip and palate,CLP）的发病危险因素。方法 对189例 CLP患儿的母亲及60例非CLP患儿的母亲进行成组病例对照研究,调查内容包括可能引起CLP发生的危险因素,用非条件Lo-gistic回归方法进行单因素和多因素分析。结果 单因素分析显示与唇裂伴或不伴腭裂（CL ± P）发病有显著性意义的因素共8种,最终进入多因素Logistic回归模型的变量有家族史、O型血、出生季节（5月～7月）和母亲职业:与腭裂（CP）发病有显著性意义的因素共4种,进入多因素 Logistic回归模型的变量为胎次、出生季节（11月～次年1月）和母亲职业。结论 环境和遗传因素是CL±P的主要发病因素,而CP则主要与环境因素有关。     

MSX1基因A274V多态性与非综合征性唇腭裂的关系

目的 研究同源异型盒基因1 (muscle segment homeobox1,MSX1) A274V多态性与非综合征性唇腭裂(non-syndromic cleft lip with or without cleft palate,NSCL/P)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,在162例非...     

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。方法：采用聚合酶链反应-限制性片段长度多态性检测法对200例NSCL/P患者和213例健康人进行基因型检测。结果：MTHFR C677T对照组与病例组在基因型分布无统计学差异（P〉0.05）,TT基因型和携带T等位基因儿童罹患NSCL/P的风险分别是CC基因型儿童的1.84倍及1.57倍。进一步分层分析发现TT基因型和CT基因型能分别显著增加儿童唇裂伴或不伴腭裂和单纯性唇裂的发病风险。MTHFR A1298C病例组和对照组在基因型频率和等位基因频率有统计学差异（P〈0.05）,AC基因型和携带C等位基因的儿童罹患NSCL/P的风险分别比AA基因型儿童降低49%及43%。分层分析中,AC基因型和携带C等位基因可降低罹患唇裂伴腭裂及唇裂伴或不伴腭裂的风险。结论：MTHFR C677T可能为中国江苏地区汉族儿童NSCL/P的危险因素,而MTHFR A1298C有可能是NSCL/P发生的保护因素。     

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.     

MSX1基因多态性与山西地区非综合征性唇腭裂的相关性研究

目的:研究MSX1基因外显子区rs13127820(M146L)、rs62636562(A274V)位点多态性与非综合征性唇腭裂(non-syn-dromic 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方法,检测243例非综合征性唇腭裂(NSCL/P)患者和292例正常对照者全血标本中MSX1基因rs13127820(M146L)、rs62636562(A274V)位点的多态性。结果:MSX1基因位点rs13127820(M146L)、rs62636562(A274V)基因型频率分布符合Hardy-Weinberg平衡;MSX1基因位点rs13127820(M146L)、rs62636562(A274V)等位基因频率分布在NSCL/P组与对照组之间差异有统计学意义(P<0.05)。结论:结果显示MSX1基因位点多态性与NSCL/P的发生有关。     

BMP4、TGF-β3基因多态性与环境暴露在非综合征性唇腭裂发生中的交互作用

目的 探讨环境暴露因素、骨形态生成蛋白4(BMP4)基因、转化生长因子β3(transforming growth factor beta-3,TGF-β3)基因之间的交互作用在非综合征性唇腭裂(nonsyndromic cleft lip and cleft palate,NSCLP)发生中的可能作用.方法 通过问卷调查获取环境暴露资料.用聚合酶链反应(PCR)-限制性片段长度多态性(restriction fragment length polymorphism,RFLP)技术对对照组(200例)和NSCLP组(200例)各基因位点的多态性进行检测.采用多因子降维法(multifactor djmensionality reduction,MDR)分析基因之间、基因与环境之间的交互作用关系,并对筛选的交互作用关系用Logistic回归进行验证.结果 BMP4 T538C、TGF-β3 C641A和TGF-β3G15572-三个单核苷酸多态性(single nucleotide polymorphism,SNP)位点间的交互作用与NSCLP的发生无关联.基因与环境交互作用分析发现,BMP4 T538C与母亲妊娠早期被动吸烟、母亲妊娠早期感染史对NSCLP的发生具有交互作用;TGF-β3G15572-与母亲妊娠早期被动吸烟、母亲妊娠早期感染史、父亲知晓妊娠前吸烟、父亲知晓妊娠前饮酒、母亲妊娠早期补充维生素对NSCLP的发生具有交互作用.经Logistic回归验证,结果一致.结论 NSCLP是基因与环境因素共同作用的结果,易感基因多态性影响着个体对环境因素的反应,研究它们之间的相互关系对阐明NSCLP的病因及发病机制具有重要意义.     

Linkage disequilibrium between MSX1 and non-syndromic cleft lip/palate in the Chilean population

Non-syndromic cleft lip/palate (NSCLP) is a complex genetic trait. Linkage and association studies have suggested that a clefting locus could be located on chromosome 4p. Sixty Chilean families were recruited for this study; from these, we used unrelated trios to evaluate the possible linkage disequilibrium between MSX1 and NSCLP. An intragenic marker, MSX1-CA, and an extragenic marker, D4S432 at a distance of 0.8 cM from MSX1, were analyzed by means of polymerase chain-reaction with fluorescent-labeled forward primers, followed by electrophoresis on a laser-fluorescent sequencer. We carried out a transmission/disequilibrium test (TDT) for multiple alleles to evaluate the presence of linkage disequilibrium. Results showed a preferential transmission of the 169-bp allele of MSX1 (p = 0.03). Although there was no preferential transmission for the D4S432 marker, the overall extended TDT (ETDT) showed a significant result (p = 0.01). The authors' findings support the hypothesis of the contribution of MSX1 in the etiology of NSCLP in the Chilean population.     

Association of MSX1 799 G>T variant with nonsyndromic cleft lip/palate in South Indian adolescent patients

International Journal of Paediatric Dentistry 2012; 22: 228–231 Background. Nonsyndromic cleft lip/palate (NSCLP) is a common congenital anomaly with significant medical, psychological, social, and economic ramifications. It is an example of complex genetic trait. There is sufficient evidence to hypothesise that disease locus for this condition can be identified by candidate genes. The purpose of this study was to test whether MSX1 (799 G>T) gene variant was involved in the aetiology of NSCLP. Methods. Blood samples were collected with informed consent from 25 subjects having NSCLP and 25 controls. Genomic DNA was extracted from the blood samples, polymerase chain reaction was performed (PCR), and digestion products were evaluated. Results. The Results showed a positive correlation between MSX1 (799 G>T) gene variant and NSCLP patients. Conclusion. MSX1 (799 G>T) gene variants may be a good screening marker for NSCLP.     

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.     

No evidence for linkage and association between 4q microsatellite markers and nonsyndromic cleft lip and palate in chilean case-parents trios.

OBJECTIVE: Nonsyndromic cleft lip/palate (NSCLP) has the characteristics of a complex genetic trait. Linkage and association studies have suggested that one or more clefting loci may be located on chromosome 4q. The goal of this study was to evaluate the possible linkage and association due to linkage disequilibrium between five microsatellite markers located on 4q28 to 4q33 and NSCLP, using the case-parent trio design. SUBJECTS AND METHODS: A total of 56 Chilean families (32 simplex and 24 multiplex) were recruited. Microsatellite markers were analyzed using polymerase chain reaction with fluorescent-labeled forward primers, followed by electrophoresis on a laser-fluorescent sequencer. Case-parents trios were ascertained to assess linkage and linkage disequilibrium through a multistage procedure. Transmission disequilibrium tests for multiple alleles were carried out to assess the statistical significance of 4q28 to 4q33 microsatellite markers. RESULTS: Only weak evidence for linkage was obtained for the FGA marker (asymptotic uncorrected p value = .08 and empirical p value = .05). Only the FGA and UCP1 markers were selected for association analysis in trios, with unrelated cases achieving a nearly significant result for the UCP1 marker (asymptotic uncorrected p value = .07 and empirical p value = .19). CONCLUSION: Though the FGA and UCP1 markers showed nearly significant p values for linkage and association, respectively, the results of the present study provided insufficient evidence of the existence of a major susceptibility locus in the 4q region that was analyzed in the present study.     

Association Between PAX9 Single-Nucleotide Polymorphisms and Nonsyndromic Cleft Lip With or Without Cleft Palate

ABSTRACT: The purpose of this study was to investigate the contribution of PAX9 gene to the risk of nonsyndromic cleft lip with or without cleft palate (NS-CL/P). The samples consisted of 142 Korean NS-CL/P families (90 males and 52 females; 9 cleft lip, 26 cleft lip and alveolus, and 107 cleft lip and palate; 76 trios and 66 dyads). A total of 10 single-nucleotide polymorphisms (SNPs) were tested for association with Korean CL/P case-parent trios using transmission disequilibrium test (TDT) and conditional logistic regression models. The minor allele frequency, heterozygosity, and a χ test for Hardy-Weinberg equilibrium at each SNP were computed between parents. Pairwise linkage disequilibrium was computed as both D' and r for all SNPs. Both allelic and genotypic TDTs were performed for individual SNPs using family-based association test program. Sliding windows of haplotypes consisting of 2 to 8 SNPs were tested using haplotype-based association test program. Genotypic odd ratios were obtained from conditional logistic regression models using STATA software. The family-based TDT using individual SNPs and 2- to 8-SNP haplotypes of the gene indicated a significant association at rs17104928 (P = 0.014). The haplotype analysis revealed that the association was most significant for the haplotype consisting of 3 SNPs (rs2073247, rs17104928, and rs17176643; P = 0.007). G/A heterozygote at rs17104928 had a significantly increased association with NS-CL/P (genotypic odd ratio, 2.88; 95% confidence interval, 1.42-5.84; P = 0.0014, dominant model). The high-risk SNP and genotype may provide a better understanding of the etiologic role of PAX9 gene in NS-CL/P and potential options for genetic counseling.     

小泛素化修饰基因-1多态性与非综合征型唇腭裂的相关性研究

目的探讨小泛素化修饰基因-1（SUMO-1）rs6709162、rs7599810和rs7580433位点单核苷酸多态性与非综合征型唇腭裂（NSOC）的相关性。方法收集宁夏地区NSOC患者208例、患者父亲189例、患者母亲176例、完整核心家系（患者及其父母）172个进行研究,并收集正常新生儿284例作为对照。采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法检测SUMO-1基因多态位点rs6709162、rs7599810和rs7580433基因型,并进行病例对照分析、传递不平衡检验（TDT）和以家系为基础的相关性检验（FBAT）。结果病例对照研究发现：SUMO-1基因rs7599810位点的TT基因型频率在唇裂、腭裂组与对照组比较有统计学差异（P=0.01,P=0.01）。TDT分析结果：rs7599810位点的T等位基因在唇腭裂组中存在过传递（P=0.00）;rs6709162位点的C等位基因在腭裂和唇腭裂组中存在过传递（P=0.00,P=0.01）;rs7580433位点的G等位基因在唇裂组中存在过传递（P=0.05）。FBAT分析结果：rs7599810位点TT基因型和T等位基因的分布具有统计学意义（P=0.00,P=0.00）。结论SUMO-1基因多态性与NSOC存在相关性。     

基于核心家系的黏附连接相关蛋白1基因多态性与广西白裤瑶族儿童龋病遗传易感性关联分析

目的 探讨黏附连接相关蛋白1(adherens junctional associated protien 1,AJAP1)基因多态性与广西白裤瑶族儿童龋病遗传易感性之间的关联。方法 选取2018年广西河池市南丹县里湖瑶乡12岁白裤瑶族高龋儿童（30人）及其生物学父母（60人）组成的三人核心家系为研究对象。利用SNPscan技术对AJAP1基因rs3896439、rs4654438位点进行分型,通过FBAT 2.0.4软件进行基于核心家系的传递不平衡检验（transmission disequilibrium test,TDT）及单倍型关联分析。结果 TDT分析结果显示,rs3896439位点单核苷酸多态性与龋病的遗传易感性显著相关,G等位基因从杂合子父母向高龋子代过度传递,为龋病的危险因素（加性模型：Z=2.263,P=0.024;显性模型：Z=2.064,P=0.039）。单倍型关联分析结果显示,rs3896439-rs4654438的单倍型G-C从杂合子父母向高龋子代显著过度传递,与龋病的遗传易感性相关联（加性模型：Z=2.180,P=0.029;显性模型：Z=2.206,P=0...     

Association of MSX1 with nonsyndromic cleft lip and palate in a Colombian population.

OBJECTIVE: To evaluate the association between MSX1 CA polymorphism and nonsyndromic cleft lip with or without cleft palate (CL+/-P) in a group of patients from Operation Smile Colombia. DESIGN: Four alleles from MSX1 CA microsatellite sequence were analyzed. Polymerase chain reaction was carried out. The amplifications were performed by Short Tandem Repeats (STRs) in ABI PRISM 310 genetic analyzer. Chi-square and odds ratio tests were used to determine the association between genotype frequencies and the risk to the cleft lip/palate in a Colombian group population. SETTING: Operation Smile Colombia. PARTICIPANTS: Ninety-four affected patients (49 men and 45 women with CL+/-P) and 93 control individuals (43 men and 50 women). RESULTS: A significant statistical difference (p<.0106) was found between the patients who carried allele 3 and CL+/-P. In addition, allele 4 (heterozygous and homozygous form) was the most frequent in CL+/-P (74%) patients and in the control group (82%). CONCLUSIONS: These findings show a positive association between the MSX1 CA polymorphism and CL+/-P in a Colombian group population.     

Association of Wnt3A gene variants with non-syndromic cleft lip with or without cleft palate in Chinese population

Objective

Non-syndromic cleft lip with or without cleft palate (NSCLP) is one of the most common birth defects all over the world. Both genetic and environmental factors may contribute to NSCLP. Recent studies have demonstrated that Wnt/β-catenin signalling pathway is required for lip and palate formation. WNT family may play an important role in the development of NSCLP. This study aimed to evaluate the association between Wnt3A gene polymorphisms and NSCLP in Chinese population from Northwest China.

Design

216 patients with NSCLP and 233 normal controls were genotyped for two SNPs of Wnt3A by PCR-RFLP. Both SNPs genotype frequencies were analysed between cases group and controls group.

Results

The frequencies of rs752107 TT and rs3121310 AA were significantly higher in NSCLP cases group (7.4%, 15.3%) than that in controls group (2.1%, 9.5%) with p-value = 0.013, 0.014, corrected p value (p-corr) <0.05 and with odds ratio (OR) = 3.49, 95% confidence interval [CI]: 1.244–9.79, OR = 2.27, 95% CI: 1.17–4.38, respectively; the frequency of rs3121310 GA was also higher in NSCLP cases group (57.4%) than in controls group (52.0%) with p-value = 0.042 and OR = 1.56 (95% CI: 1.02–2.39). And the frequency of rs752107 TT of Wnt3A showed higher risk in female patients, while the frequency of A allele of rs3121310 showed stronger association in male patients.

Conclusions

This is the first report that two SNPs of Wnt3A (rs752107 and rs3121310) are significantly associated with NSCLP in Chinese population. These findings provide a context for understanding the genetic aetiology of NSCLP.