GLI3基因与单纯性马蹄内翻足的关联分析及其突变筛查

目的对GLU基因与单纯性马蹄内翻足进行关联分析和突变筛查，探讨GLU基因与单纯性马蹄内翻足的相关性。方法应用限制性片段长度多态性分析技术，分析84个单纯性马蹄内翻足核心家系中GLI3基因内两个单核苷酸多态（single nucleotide polymorphisms, SNP）位点的基因型，并应用ETDT软伯统计分析各SNP位点基因型与单纯性马蹄内翻足的关联；应用变性梯度凝胶电泳技术对103例单纯性马蹄内翻足患者GLI3基因的第9至12外显子进行突变筛查。结果经ETDT分析，位于GLI3基因第4外显子的cSNP rs846266差异无统计学意义（χ^2=3．3582，P〉0．05）；第14外显子的cSNP rs929387差异有统计学意义（χ^2=7．2466，P〈0．05），在单纯性马蹄内翻足核心家系中存在传递不平衡；发现1例患者及其母亲的第9外显子有108（G→A）的同义点突变。结论GLI3基因与单纯性马蹄内翻足相关，其第9至12外显子可能并非该病的突变热点。     

单胺氧化酶A基因多态性与精神分裂症的关联研究

目的探讨河南地区汉族人群单胺氧化酶A（monoamine oxidase A，MAOA）基因多态性与精神分裂症的关系。方法参照CCMD-3诊断标准，选取212例精神分裂症患者与168名正常对照，应用聚合酶链反应及限制性片段长度多态性技术检测MAOA基因多态性，采用病例一对照的关联分析方法对精神分裂症患者及正常对照的基因型和等位基因频率进行分析。结果（1）MAOA基因的基因型在患者组和对照组中均符合Hardy-Weinberg平衡定律（X^2=0．618，dr：2，P〉0、05；X^2=3．173，df=2，P〉0．05）。（2）MAOA基因的基因型和等位基因频率在患者组与对照组间的分布差异无统计学意义（P〉0．05）。（3）按性别分组，男性患者组中CT基因型分布频率显著高于男性对照组（X^2=7．654，P=0．022）。（4）MAOA基因的基因型和等位基因频率在家族史阴、阳性间的分布差异无统计学意义（P〉0．05）。结论没有发现MAOA基因多态性与汉族精神分裂症的发病有关联，但对性别发病有影响，基因型CT可能是男性精神分裂症发病的易感因素。     

强直性脊柱炎患者MICA基因第2、3和4外显子的多态性及其与HLA-B抗原的连锁不平衡

目的探讨皖籍汉族人群MICA基因（major histocompatibility complex class Ⅰchain-related gene A，MICA）第2、3、4外显子的多态性，及其与HLA-B抗原的连锁不平衡在强直性脊柱炎（ankylosing spondylitis，AS）发病中的作用。方法采用聚合酶链反应-序列特异性寡核苷酸探针杂交（polymerase chain reactionsequence-specific oligonucleotide probing，PCR-SS0）技术对56例AS患者和112名正常对照人群进行MICA基因第2、3、4外显子的多态性和HLA-B抗原的检测。结果AS患者和正常对照人群的MICA等位基因分布均以MICA＊008占优势，频率分别为32．14％和30．36％。两组人群MICA＊007等位基因的分布差异有统计学意义（X^2=10．18，P〈0．05，RR=2．50）。单倍型分析显示，AS患者和正常对照人群的MICA等位基因均显示出与多个HLA-B位点的连锁不平衡现象，两组间差异有统计学意义的单倍型为MICA＊007-B27（X^2=18．46，P〈0．05，RR=7．47）。分层分析结果显示，HLA-B27阳性与AS的相关性有统计学意义（P〈0．05），但MICA＊007基因与AS的相关性无统计学意义（P〉0．05）。结论AS患者中MICA＊007等位基因频率的显著升高可能源于MICA基因与HLA-B位点间的广泛连锁不平衡。     

P基因在中国汉族及藏族群体中的多态性研究

目的：研究中国汉族和藏族两个群体中P基因3个外显子多态性。方法：利用PCR－RFLP多态性方法扩增P基因的3个外显子10、14、24,对其产物分别用HaeⅢ、MboⅡ、MaeⅢ酶切,4％琼脂糖凝胶电泳确定基因型。结果：A355A和G780G的等位基因频率在两个群中的差异具有显著性（χ^2＝24．54,χ^2=37.05,P＜0．001）,各基因型频率分布在两个群体间差异具有显著性（χ^2＝28．39,P＜0．001;χ^2=33.72,P＜0．001）;在两个群体内部男女基因型频率差异无显著性;IVS13－15的等位基因频率在两个群体中差异无显著性（χ^2＝2．06,P＜0．05）。汉族与藏族的基因型分布符合Hardy-Weinberg平衡。结论：P基因在不同的人群中具有很高的多态性。     

神经型烟碱乙酰胆碱受体α7亚单位基因多态性与精神分裂症的传递不平衡研究

目的 探讨神经型烟碱乙酰胆碱受体α7亚单位基因(neuronal nicotinic acetyleholine receptor α7 subunit gene,CHRNA7)多态性与精神分裂症的关系.方法 应用聚合酶链反应及聚丙烯酰胺凝胶芯片技术检测129个精神分裂症先证者核心家系CHRNA7基因的rs2337980、rs1909884、rs883473三个单核苷酸多态性,并采用基于单倍型的单倍型相对风险检验(haplotype relative risk,HHRR)、传递不平衡检验(transmission disequilibrium test,TDT)及单倍型分析进行统计.结果 (1)HHRR分析结果显示rs2337980位点精神分裂症患者组与虚拟对照组之间等位基因频率差异有统计学意义(P=0.017);(2)TDT分析发现,rs2337980位点与精神分裂症之间可能存在传递不平衡,杂合子父母过多的传递等位基因C给患病子女(P=0.021).(3)单倍型分析发现,rs2337980、rsl909884及rs2337980、rsl909884、rs883473组成的单倍型与精神分裂症有显著相关(总体P=0.034;glohal P=0.027),其中T-C,T-C-T两个单倍型与精神分裂症可能存在传递不平衡.结论 CHRNA7 基因多态性可能与精神分裂症存在关联,rs2337980的变异等位基因T可能是精神分裂症的保护性因子.     

G-蛋白信号转导调节子4(RGS4)基因多态性与精神分裂症的关联研究

目的：探讨G-蛋白信号转导调节子4（regulator of G-protein signaling-4，RGS4）基因与精神分裂症及临床症状的遗传关联。方法：应用病例对照关联研究设计，采用聚合酶链式反应-限制性片断长度多态性（PCR-RFLP）和DNA测序方法，分析386例精神分裂症患者和390例正常对照者中RGS4基因4个单核苷酸多态性（SNP）位点与精神分裂症的关联。并采用阳性和阴性症状量表（PANSS）评估患者的临床症状，进一步分析PANSS因子分与RGS4多态性的关联。结果：RGS4基因的两个多态性位点rs12753561（T〉G，χ^2=8．970，P=0．002）和rs10759（C〉A，χ^2=13．773，F=0．002，P=0．002）与精神分裂症关联，由上述4个SNPs组成的多个单体型如AAGA（χ^2=11．120，P=0．0008，OR=0．52，95％CI=0．36—0．77）和GGGC（χ^2=10．096，P=0．001，OR=1．43，95％CI=1．15—1．79）均与精神分裂症关联。PANSS量表的阴性症状因子分与rs12753561（t=2．216，P=0．029）和rs10759（t=2．543，P=0．012）关联。结论：RGS4基因多态性与精神分裂症及阴性和一般精神病理症状显著关联。     

DIO2基因单核苷酸多态性与精神发育迟滞相关性研究

目的探讨DIO2基因多态性及单倍型与精神发育迟滞的相关关系。方法本研究以344个汉族精神发育迟滞患者及其亲属组成的家系为研究对象．在DIO2上选择了3个合适的SNPs作为标记．利用PCR—SSCP和PCR—RFLP方法完成基因分型。应用FBAT软件对多态位点及可能组成的单倍型与精神发育迟滞的关系进行分析。结果单个位点家系关联性分析结果提示：rs225015、rs225014和rs225012位点各等位基因从亲代传递给患病子代没有观察到显著性（P〉0．05）。多态性位点间的连锁不平衡检验显示：3个位点rs225015、rs225014和rs225012均具有较高的连锁不平衡（D’〉0．8）。单倍型分析结果显示：3个位点构成的单体型GTG在附加遗传模型（Z=2．226,P=0．026019）和隐性遗传模型（Z=2．651,P=0．008023）与MR相关。结论DIO2基因可能与精神发育迟滞的易感性有关。     

湖北土家族人群HLA-A、B等位基因及单倍型多态性的分布

目的研究湖北五峰县土家族190名无亲缘关系健康个体HLA-A、B等位基因及单倍型频率的分布，为进一步研究HLA基因多态性与疾病的关联奠定基础。方法采用序列分型（sequence-baaed typing，SBT）法对最具多态性的HIA-A、B基因2、3外显子进行了基因型分析，采用Arlequin软件对群体基因、单倍型频率进行最大估计值的计算。结果HLA-A、B等位基因符合Hardy-Weinberg平衡定律（P〉0．05），无显著的连锁不平衡现象，共检测出26个HLA-A等位基因及41个HLA-B等位基因，其中频率最高的为A＊0201（0．16053），A＊110101（0．14737），A＊24020101（0．14211），B＊4001（0．14737），B＊4601（0．13947），其次为A＊0207（0．08947），A＊0206（0．08158），B＊1301（0．07632），B＊5801（0．08947），B＊1501（0．09737），而大于0．05％的为A＊330301（0．05526），B＊1502（0．05526），B＊3501（0．05263），单倍型分布较普遍的为A＊0202-B＊4001（0．04196），A＊0201-B＊4601（0．03625）。结论采用高分辨测序分型法对土家族人群HLA-A、B等位基因进行分型的实验结果可做为湖北土家族人HLA-A、B等位基因、单倍型频率的群体资料，对进一步开展群体遗传学、临床器官移植、疾病关联、HLA遗传学特征、法医学、人类学等研究具有重要意义。     

A2M基因多态性与帕金森病和特发性震颤的关联

目的探讨α2-巨球蛋白基因（α2-macmglobulin gene，A2M）第24外显子1000G／A及第18外显子5’端剪接点5个碱基插入／缺失（insertion／deletion，IZD）多态与中国北方帕金森病（Parkinson’sdisease，PD）和特发性震颤（essential tremor，ET）的关联。方法应用聚合酶链反应-限制性片段长度多态性方法，对87例PD、73例ET和100名健康对照者A2MG／A和I／D两个位点的基因型和等位基因分布频率进行检测。结果（1）A2M基因G／A位点基因型和等位基因分布，在PD与ET、对照组间差异有统计学意义（P〈0．05），PD组的G等位基因和GA基因型明显高于ET、对照组（P〈0．05）；而ET与对照组间相比，差异无统计学意义（P〉0．05）。（2）A2M基因IZD位点基因型和等位基因分布，在PD、ET、对照组间差异无统计学意义（P〉0．05）。结论（1）G／A位点多态与PD的发病有关联，G／A位点多态与ET无关。（2）I/D位点多态与中国北方PD、ET无关。     

精神分裂症与六种候选功能基因的关联研究

目的：探讨多巴胺D2受体基因（dopamine D2 receptor,DRD2)、多巴胺D4受体基因（DRD4）、5－羟色胺2A受体基因（5－hydroxytryptamine 2A receptor,5-HT2A),5-羟色胺6受体基因（5－HT6)、儿茶酚胺氧位甲基转移酶基因（catechol-O-methyltransferase,COMT)和多巴胺转运体基因（dopamine transferase,DAT1)多态性与精神分裂症的关系。方法：应用基因扩增片段长度多态和基因扩增的限制性片段长度多态技术，对中国汉族人群中67例精神分裂症患者与上述6种候选功能和基因扩增的限制性片段长度多态技术，对中国汉族人群中67例精神分裂症患者与上述6种候选功能基因进行遗传关联分析。结果：（1）DRD2、5－HT2A、5－HT6和KCOMT的基因型和等位基因频率在患者组和对照组中差异无显著性（P＞0．05）。（2）DRD4基因中6次重复序列等位基因、DAT1基因中480bp等位基因和480／520基因型在两组中差异有显著性，Z分别为2．03、2．05和2．05；P均小于0．05。（3）经关联分析后，仅DAT1基因的480bp等位基因的比值比为0．441，95％可信区间为0．202－0．963，并有显著性意义（Z＝2．05，P＜0．05），而DAT1的480／520基因型和DRD4和6次重复序列等位基因的比值比分别为0．128和0．123，但Z均小于1．96，无显著性意义（P＞0．05）。因此，6个功能基因中仅DAT1的480bp等位基因与精神分裂症呈负关联。结论：中国汉族人群中DAT1基因的480bp等位基因与精神分裂症间存在负关联，支持精神分裂症的多巴胺假说。     

The transmission disequilibrium analysis between neuronal nicotinic acetylcholine receptor alpha 7 subunit gene polymorphisms and schizophrenia]

OBJECTIVE: To investigate the association between neuronal nicotinic acetylcholine receptor alpha 7 subunit (CHRNA7) gene and schizophrenia. METHODS: The three polymorphisms rs2337980, rs1909884, rs883473 in CHRNA7 gene were detected based on PCR and polyacrylamide gel microarray in 129 schizophrenic trios. The results of genotyping were analyzed by haplotype relative risk analysis based on haplotype(HHRR), transmission disequilibrium test(TDT) and hyplotype analysis. RESULTS: (1)The HHRR analysis suggested that there was significant differences in rs2337980 allele frequencies between schizophrenia group and dummy control group(P= 0.017); (2)In TDT test, there may be transmission disequilibrium between rs2337980 and schizophrenia, the heterozygous parents excessively transferred the C allele to patients (P= 0.021); (3)The haplotype between rs2337980 and rs1909884 as well as the hyplotype among rs2337980, rs1909884 and rs883473 may have significant association with schizophrenia (global P= 0.034; global P= 0.027), the T-C and T-C-T hyplotype may have transmission disequilibrium with schizophrenia. CONCLUSION: There may be association between CHRNA7 gene polymorphisms and schizophrenia, the variant allele T in rs2337980 may have a protective effect to schizophrenia.     

Neuregulin 1基因多态性与中国汉族精神分裂症关联分析

目的 探讨Neuregulin 1(NRG1)基因多态性与精神分裂症的关联.方法 在258个中国汉族精神分裂症核心家系(患者及其亲生父母)中,应用实时定量PCR技术检测位于NRGl基因5'端的4个单核苷酸多态(single nucleotide polymorphism,SNP)位点:rs221533(C/T)、rs7820838(C/T)、433E1006 (A/G)和rs3924999(C/T),进行基因分型,应用传递不平衡检测(transmission disequilibrium test,TDT)分析等位基因传递情况,分析该基因与精神分裂症易感性的关联.结果 在258个中国汉族核心家系中,rs221533、433E1006、rs3924999三个SNP均存在有统计学意义的传递不平衡,优先传递的等位基因分别是:C、A、T(rs221533:X2=27.45,P=0.000;433E1006:X2=56.08,P=0.000;rs3924999:X2=10.53,P=0.001).rs7820838未检到不平衡传递(X2=3.31,P=0.081).频率大于1%单倍型进行分析,rs221533-rs7820838-433E1006联合分析,单倍型C/C/G和C/C/A优先传递(C/C/G:X2=5.26,P=45.08;C/C/A:X2=0.026,P=0.000);rs221533-rs7820838-433E1006-rs3924999联合分析,单倍型C/C/G/T、C/C/A/C和C/C/A/T优势传递(C/C/G/T:X2=10.71,P=0.001;C/C/A/C:.)X2=8.83,P=0.006、C/C/A/T:X2=27.00,P=0.000).213个阳性亚型的精神分裂症核心家系中传递不平衡得出基本一致的结果 .结论 Nrg1基因多态性与中国汉族人群精神分裂症存在关联,尤其是支持与阳性亚型精神分裂症存在关联.     

Xp1区内定位注意缺损多动障碍易感位点的研究

目的探讨注意缺损多动障碍(attention deficit hyperactivity disorder, ADHD)与单胺氧化酶(monoamine oxidase, MAO)A型基因的遗传关系。方法采用基于单体型相对风险(haplotype-based haplotype relative risk,HHRR)和传递不平衡检验(transmission disequilibrium test, TDT)的方法,在60个ADHD儿童和双亲中进行了MAOCA微卫星多态性的关联和连锁分析。结果经HHRR和TDT分析,ADHD儿童与MAOCA位点114 bp相关联和连锁(χ2分别为4.90和4.84,P＜0.05)。结论 ADHD与MAO A型基因相关联和连锁,其易感位点可能定位于Xp1区。     

No association between 5HT-2A and bipolar disorder irrespective of genomic imprinting.

Recent evidence that 5HT-2A may be subjected to genomic imprinting prompted us to examine a collection of Irish family trios (an affected individual and both parents) for evidence of an association between 5HT-2A and bipolar disorder. Family trios offer an advantage over case control studies in regard to genomic imprinting since with family trios it is possible to trace the path of alleles from the parents to the offspring. Using haplotype-based haplotype relative risk (HHRR) and transmission/disequilibrium (TDT) analyses, no evidence was found for an association of 5HT-2A with bipolar affective disorder under the assumption of no imprinting and of imprinting.     

VEGF C-634G polymorphism is associated with protection from isolated ventricular septal defect: case-control and TDT studies

The ventricular septal defect (VSD) is the most common congenital heart defect and no candidate susceptibility gene has been identified. Endocardial cushion and outflow septal morphogenesis, malalignment of which induces VSD, have been suggested to be mediated by the vascular endothelial growth factor (VEGF). Three single-nucleotide polymorphism (SNP) variants in promoter and 5'-UTR region of the VEGF gene, C-2578A (rs699947), G-1154A (rs1570360) and G-634C (rs2010963), were reported to alter its expression. We assessed the association in a Chinese population between these SNPs and VSD using a double approach: case-control and TDT designs. Among the three SNPs, only -634C allele was less frequently present in 222 patients compared to 352 controls (odds ratio: 0.76, 95% CI: 0.59-0.97, X(2)=5.06, P=0.024, not significant after a Bonferroni correction). This was significantly less transmitted to VSD patients (trios: 142) (odds ratio: 0.39, 95% CI: 0.25-0.62, X(2)=8.11, df=1, P=0.004, corrected P=0.024). A similar result was observed for haplotype -2578C/-1154G/-634C allele in both studies (in TDT: X(2)=7.51, df=1, P=0.006, corrected P=0.048). All these associations for the first time demonstrated that -634C allele was in a significant protective association against VSD, suggesting that VEGF dysregulation was involved in the pathological processes of VSD.     

No evidence for involvement of KCNN3 (hSKCa3) potassium channel gene in familial and isolated cases of schizophrenia

Several studies have reported in schizophrenia a decrease of age of onset in successive family generations, and this observation is consistent with anticipation. Anticipation is known to result from expansion of CAG repeats in several neurodegenerative disorders. Longer alleles of the KCNN3 gene, which contains a highly polymorphic CAG repeat, and encodes a neuronal small conductance calcium-activated potassium channel, have recently been shown to be over-represented in sporadic cases of schizophrenia. In this report, we tested the hypothesis of an association between longer alleles of CAG repeat in the KCNN3 gene and schizophrenia in 20 families with clinical evidence for anticipation and in 151 unrelated schizophrenic cases. No significant difference in the distributions of allele frequencies was observed between familial cases of schizophrenia and controls, and between unrelated cases and controls. Furthermore, no intergenerational CAG repeat instability was detected in the 20 families. Our results do not support the involvement of the KCNN3 (hSKCa3) gene in the etiology of schizophrenia.     

Enhanced APOE2 transmission rates in families with autistic probands

We have previously described linkage/association between reelin gene polymorphisms and autistic disorder. APOE also participates in the Reelin signaling pathway, by competitively antagonizing Reelin binding to APOE receptor 2 and to very-low-density lipoprotein receptors. The APOE2 protein variant displays the lowest receptor binding affinity compared with APOE3 and APOE4. In this study, we assess linkage/association between primary autism and APOE alleles in 223 complete trios, from 119 simplex Italian families and 44 simplex and 29 multiplex Caucasian-American families. Statistically significant disequilibrium favors the transmission of epsilon2 alleles to autistic offspring, over epsilon3 and epsilon4 (allele-wise transmission/disequilibrium test [TDT], chi2 = 6.16, 2 degrees of freedom [d.f.], P<0.05; genotype-wise TDT, chi2 = 10.68, 3 d.f., P<0.05). A novel epsilon3r allele was also discovered in an autistic child and his mother. Autistic patients do not differ significantly from unaffected siblings (allele-wise TDT comparing autistic patients versus unaffected sibs, chi2 = 1.83, 2 d.f., P<0.40, not significant). The major limitation of this study consists of our small sample size of trios including one unaffected sibling, currently not possessing the statistical power necessary to conclusively discriminate a specific association of epsilon2 with autism, from a distorted segregation pattern characterized by enhanced epsilon2 transmission rates both to affected and unaffected offspring. Our findings are thus compatible with either (a) pathogenetic contributions by epsilon2 alleles to autism spectrum vulnerability, requiring additional environmental and/or genetic factors to yield an autistic syndrome, and/or (b) a protective effect of epsilon2 alleles against the enhanced risk of miscarriage and infertility previously described among parents of autistic children.     

No association between 5HT‐2A and bipolar disorder irrespective of genomic imprinting

Recent evidence that 5HT‐2A may be subjected to genomic imprinting prompted us to examine a collection of Irish family trios (an affected individual and both parents) for evidence of an association between 5HT‐2A and bipolar disorder. Family trios offer an advantage over case control studies in regard to genomic imprinting since with family trios it is possible to trace the path of alleles from the parents to the offspring. Using haplotype‐based haplotype relative risk (HHRR) and transmission/disequilibrium (TDT) analyses, no evidence was found for an association of 5HT‐2A with bipolar affective disorder under the assumption of no imprinting and of imprinting. © 2001 Wiley‐Liss, Inc.     

hSKCa3: no association of the polymorphic CAG repeat with bipolar affective disorder and schizophrenia

hSKCa3 is a neuronal small conductance calcium-activated potassium channel, which contains a polyglutamine tract, encoded by a polymorphic CAG repeat in the gene. Since an association between longer alleles of this CAG repeat and bipolar disorder or schizophrenia has been reported, we genotyped the polymorphic CAG repeat in 91 German family trios of patients with bipolar disorder I and used the transmission disequilibrium test (TDT) to test for association. Applying a dichotomized model (< or = 19 or > 19 CAG triplets), we found no evidence for an association of longer alleles with bipolar disorder (TDT = 0.75, P = 0.386). Regarding the whole range of alleles, there was no preference in transmitting the larger of the two observed alleles from parents to the affected offspring. In parallel we performed an independent case-control study on German patients with bipolar disorder and schizophrenia. Again we did not detect an overrepresentation of longer CAG repeats in patients. Thus, our data do not support the hypothesis that longer CAG repeats in the hSkCa3 gene contribute to the susceptibility for bipolar disorder and schizophrenia.