先天性心脏病核心家庭血清同型半胱氨酸水平及相关基因多态性研究

目的:了解MTHFR C677T、MS A2756G、MTHFD G1958A和CBS 844 ins68bp位点在中国北方正常人群中的基因型分布,并评价单一或复合位点基因变异与叶酸、维生素B12、同型半胱氨酸(Hcy)水平及先天性心胖病(CHD)的关系.方法:选择辽宁省192例CHD患者及其父母作为病例组,同一地区年龄、性别匹配的124名正常人及其父母作为对照组,采用PCR-RFLP方法检测其基因型,放射免疫法和荧光偏振免疫法测定血清叶酸、维生素B12和Hcy水平,比较两组差异.结果:中国北方正常人群中,这四个位点的突变等位基因频率分别为MTHFR 51.18%, MS 7.58%, MTHFD 24.32%, CBS插入频率 2.36%;CBS 844 ins68bp位点杂合型频率病例组明显高于对照组(子代为12.57% 和 2.97%, 父亲为10.88% 和 3.09%,母亲为11.54% 和 1.02%),子代的OR值为4.70 (95% CI 1.34～25.15),父亲的OR值为3.83 (95% CI 1.05～20.98),母亲的OR值为12.65 (95% CI 1.92～532.47),其他三个位点两组差异无统计学意义;MTHFR、CBS和MTHFD三个位点联合基因变异的母亲、MTHFR 和 CBS两个位点联合基因变异的母亲(OR=8.44,95%CI:1.23～362.26)、MTHFD 和 CBS两个位点联合基因变异的母亲在病例组中所占的比例明显高于对照组;病例组父亲和母亲的血清叶酸水平明显高于对照组;病例组母亲的血清Hcy水平高于对照组,但差异无统计学意义;MTHFR 和 MTHFD两个位点为纯合型者的血清叶酸和维生素B12水平轻微下降,而血清Hcy水平轻微上升;联合基因变异使血清叶酸、维生素B12和Hcy水平出现下降趋势.结论:这四个位点的基因多态性存在着种族和地区差异,CBS 844 ins68bp位点突变可能是CHD发生的一个危险因素,亲代(尤其是母亲)的插入突变可使其后代发生CHD的危险性升高.     

MTR、MTRR基因多态性与先天性心脏病的相关性研究

目的分析母子甲硫氨酸合成酶(methionine synthetas,MTR)、甲硫氨酸合成还原酶(methionine synthase reductase,MTRR)基因突变与子代先天性心脏病(congenital heart disease,CHD)发生的相关性,进一步探讨CHD的发生机制。方法通过母子配对,采取病例对照方式,选择CHD患儿及其母亲60对,正常儿童及其母亲60对,采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法检测MTR 2 756位点以及MTRR 66位点的基因型,分析基因变异与CHD发生的相关性。结果①子代及母亲CHD病例组与对照组MTR基因2 756位点基因型频率分布及等位基因频率差异均无统计学意义(P>0.05)。②CHD患儿与对照儿童的MTRR 66基因型分布比较以及患儿母亲与对照组母亲基因型分布比较,差异有统计学意义(P<0.05),2组等位基因频率比较,子代之间差异无统计学意义(P>0.05),但母亲之间差异有统计学意义(P<0.01)。③儿童与母亲MTR 2756和MTRR 66位点基因型之间的差异无统计学意义(P>0.05)。④MTRR和MTR位点均发生基...     

同型半胱氨酸代谢酶MTHFR、CBS、MS基因多态性与脑梗死患者颈动脉狭窄的相关性

目的 探讨同型半胱氨酸(Hcy)代谢酶亚甲基四氢叶酸还原酶( MTHFR)基因C677T、胱硫醚β-合成酶(CBS)基因844ins68、T27796C和甲硫氨酸合成酶(MS)基因A2756G基因多态性与脑梗死患者颈动脉狭窄的相关性.方法 脑梗死组135例,对照组65例,采用聚合酶链反应一限制性内切酶片段长度多态性技术(PCR-RFLP)检测基因表型.结果 ①脑梗死组MTHFR C677T突变(TT)基因型及T等位基因频率显著高于对照组(均P＜0.05);脑梗死伴中重度颈动脉狭窄组MTHFR C677T突变(TT)基因型及T等住基因频率高于脑梗死伴轻度颈动脉狭窄组和脑梗死无颈动脉狭窄组(均P＜0.05).②脑梗死组CBS T27796C突变(CC)基因型及C等住基因频率显著高于对照组(均P＜0.05);脑梗死伴中重度颈动脉狭窄组T27796C突变(CC)基因型及C等位基因频率高于脑梗死伴轻度颈动脉狭窄组和脑梗死无颈动脉狭窄组(均P ＜0.05).CBS844ins68位点多态性,未发现突变.③脑梗死组MS A2756G突变(AG)基因型及G等位基因频率与对照组比较无显著性差异(均P ＞0.05).脑梗死伴中重度狭窄组MS A2756G突变(AG)基因型及G等位基因频率与脑梗死伴轻度颈动脉狭窄组和脑梗死无颈动脉狭窄组比较均无显著性差异(均P ＞0.05).结论 MTHFR基因C677T位点、CBS基因T27796C位点突变与脑梗死颈动脉狭窄及其程度相关,MS基因A2756G位点突变与脑梗死颈动脉狭窄及其程度无关.     

甲硫氨酸合成酶基因A2756G多态性与2型糖尿病合并脑梗死的相关性

目的研究甲硫氨酸合成酶(MS)基因A2756G多态性与2型糖尿病合并脑梗死的相关性。方法采用聚合酶链反应—限制性片段长度多态性(PCR-RFLP)方法鉴定安徽地区562例汉族人MS基因A2756G多态性基因型,比较对照组、2型糖尿病组、2型糖尿病合并脑梗死组各组间不同基因型及等位基因频率。结果安徽地区汉族人群存在MS基因A2756G多态性,其G等位基因频率与西方人群比较明显降低,2型糖尿病合并脑梗死组的MS不同基因型及等位基因频率与对照组比较,差异无显著性(P>0.05)。结论MS基因A2756G多态性与安徽地区汉族人群2型糖尿病合并脑梗死的遗传易感性无关。     

广西壮族老年缺血性脑梗死患者蛋氨酸合成酶基因A2756G突变频率检测

目的初步探讨蛋氨酸合成酶(MS)基因A2756G突变与广西壮族老年患者缺血性脑梗死的相关性。方法运用聚合酶链反应—限制性片段长度多态性(PCR-RFLP)技术检测64例广西壮族老年患者缺血性脑梗死患者(病例组)及30例正常体检者(对照组)MS基因A2756G突变频率。结果病例组A2756G A/A、A/G及G/G基因型频率分别为82.81%、15.63%及1.56%,对照组分别为86.67%、13.33%及0,病例组A和G等位基因频率分别为90.63%和9.37%,对照组A及G等位基因频率分别为93.33%及6.67%,2组基因型频率及等位基因频率差异均无统计学意义(P>0.05)。结论 MS基因A2756G突变与广西壮族老年缺血性脑梗死的发病无明显相关,该突变可能不足以构成广西壮族老年缺血性脑梗死发病的独立危险因素。     

MTHFR基因多态性与胎儿先天性心脏畸形的关系

目的 研究探讨怀有先天性心脏病胎儿的孕妇叶酸代谢通路基因的多态性,及其与先天性心脏病(congenital heart disease,CHD)易感性之间的关系。 方法 按照纳入标准收集产前诊断B超彩色多普勒心动图诊断先天性心脏病320例,抽取外周静脉血,采用PCR-RELP方法检测MTHFR677位点和MTHFR1298位点的基因多态性。 结果 ①基因位点多态性:对照组和病例组的C和T等位基因、A和C等位基因,2组差异均有统计学意义(χ2=16.589,P<0.001;χ2=5.078,P=0.020)。②基因型多态性:对照组和病例组的CC、CT、TT基因型,2组差异有统计学意义(χ2=15.282,P<0.001),对照组和病例组的AA、AC、CC基因型2组差异无统计学意义(χ2=5.092,P=0.080)。③分层研究中:在法洛四联症小组,病例组的CC、CT、TT基因型及AA、AC、CC基因型同对照组差异均存在统计学意义(χ2=7.794,P=0.020;χ2=8.998,P=0.010);在室间隔缺损小组,病例组的CC、CT、TT 基因型同对照组差异存在统计学意义(χ2=10.407,P<0.001),而AA、AC、CC基因型同对照组差异无统计学意义(χ2=0.667,P=0.720)。 结论 母亲MTHFR C677T多态性与子代先心病发生相关,且分层研究中提示母亲MTHFR C677T多态性与子代法洛四联症、VSD发生相关。另研究结果示病例组和对照组MTHFR A1298C基因型虽然无明显差异,但等位基因A/C差异有统计学意义,提示等位基因A/C为子代发生CHD的一个危险因素。      

慢性阻塞性肺疾病TNFA和LTA基因等位基因变异分析

目的 研究中国北方汉族人群中TNF超家族基因等位基因变异是否与慢性阻塞性肺疾病(COPD)相关联.方法 以50例COPD患者为研究对象,应用聚合酶链反应一限制性片段长度多态性(PCR-RFLP)方法研究TNF超家族基因(TNFA和LTA)等位基因变异分布.结果 COPD组TNFA基因多态性位点-308G/A AA基因型频率明显高于对照组(P＜0.01),OR值为10.756(95%CI为9.875～12.640);COPD组-308G/A多态性位点A等位基因频率明显高于对照组(P＜0.01);COPD组LTA基因 252A/G多态性位点AG基因型频率明显高于对照组(P＜0.01),OR值为4.373(95%CI为3.301～6.872).COPD组TNFA基因GG正常基因型和LTA基因AG杂合基因型结合个体频率明显高于对照组(P＜0.05).2组间LTA基因 252 A/G多态性位点G等位基因频率差异无统计学意义(P＞0.05).结论 中国北方汉族人群m基因等位基因变异、TNFA基因多态性变异组合与COPD相关联.     

SHMT1基因启动子的rs638416位点与先天性心脏病的相关性

 目的  通过研究丝氨酸羟甲基转移酶(serine hydroxy methyl transferase,SHMT1)基因启动子区的单核苷酸多态性(single nucleotide polymorphism,SNP)来探讨该基因与先天性心脏病 (congenital heart disease,CHD)的相关性。方法  选取来自山东省的201例CHD患儿(病例组)和192例正常儿童(对照组),采用Sanger测序法对rs638416和rs117940726两个位点的基因分型进行病例-对照研究,进一步用双荧光素酶报告基因分析其是否为功能性SNP。结果  两个SNP位点的基因型分布在病例组和对照组中均符合Hardy-Weinberg平衡;单位点分析显示rs638416位点G等位基因在病例组中分布频率显著高于对照组 (P=0.009)。经非条件Logistic回归分析确定,Log-additive模型最适用于分析rs638416,且该模型下统计学差异最显著 (OR=1.49,95%CI:1.11～ 2.01,P=0.007 4);双荧光素酶报告基因分析显示带有G等位基因的启动子序列转录效率较带有C等位基因的启动子序列低36% (P=0.013)。结论  rs638416位点能影响SHMT1转录效率,并与心脏发育异常相关,提示rs638416位点G等位基因是山东人群CHD发生的遗传风险因子。     

冠心病和深静脉血栓患者蛋氨酸合成酶基因A2756G多态性检测

目的:检测冠心病(CHD)及深静脉血栓(DVT)形成患者蛋氨酸合成酶(MS)基因A2756G的多态性.方法:运用PCR-限制性片段长度多态性(RFLP)对103例DVT患者、208例CHD患者和250例健康对照进行MS基因A2756G多态性分析.结果:DVT组A/G G/G型频率为10.7%,低于对照组的19.6%(x2=4.114,P＜0.05),但2组等位基因频率差异无统计学意义(P＞0.05).结论:A/G G/G型可能对静脉血栓的形成具有保护性作用,该型的分布存在种族和地域差异.MS基因A2756G多态可能不是河南汉族人群CHD发生的遗传危险因素.     

脑钠尿肽基因rs198389单核苷酸多态性与冠心病的相关性研究

目的探讨脑钠尿肽(BNP)基因rs198389位点单核苷酸多态性(SNP)与冠心病(CHD)的关系。方法取病例-对照方法,收集冠心病和健康者各961例,应用Real-timeqPCR(TaqMan探针)技术进行基因分型,分析BNP基因rs198389位点SNP与CHD危险性的关系。结果 CHD组和对照组比较,AA和GG基因型、A等位基因和G等位基因分布差异有统计学意义(P<0.05);GG基因型和G等位基因频率在病例组(3.6%和16.7%)明显高于对照组(2.1%和14.3%)。与A等位基因比较,G等位基因使CHD危险性显著增加(OR=1.20,95%CI=1.01~1.43,P=0.04)。GG基因型者CHD危险性增加(P=0.04),但在调整年龄、性别、吸烟、饮酒、体重指数、血甘油三脂、高血压史、糖尿病史和CHD家族史等变量后结果差异无统计学意义(P=0.12)。结论 BNP基因位点rs198389的G等位基因使CHD危险性显著增加;BNP基因rs198389位点SNP可能与CHD易感性相关,但不是其独立危险因素。     

Polymorphism of methionine synthase gene in nuclear families of congenital heart disease

Objective To investigate the relation of methionine synthase (MS) gene variation with congenital heart disease (CHD) phenotype. Methods One hundred and ninety three CHD patients (94 males and 99 females) and their biological parents (nuclear families) in Liaoning Province were selected as the case group, and another 104 normal persons (60 males and 44 females) and their parents without family history of birth defects as the control group. For all subjects the polymorphism of MS gene A2756G locus was examined by PCR-RFLP method. Results In offspring of the control group the frequencies of MS genotype ( / -) and allele ( ) were 10.7% and 5.3%, without existence of homozygote. The MS genotype distribution and allele frequencies of CHD patients and their mothers were not significantly different from the control (P > 0.05). The frequency of allele ( ) in case fathers (5.0 %) was apparently lower than that in the control (9.1%, P=0.060), and the odds ratio (OR) was 0.53 (95% CI: 0.25-1.09). There was no diffe     

Relationship between polymorphism of methylenetetrahydrofolate dehydrogenase and congenital heart defect

Objective To investigate the relationship between G1958A gene polymorphism of methylenetetrahydrofolate dehydrogenase (MTHFD) and occurrence of congenital heart disease (CHD) in North China. Methods One hundred and ninety-two CHD patients and their parents were included in this study as case group in Liaoning Province by birth defect registration cards, and 124 healthy subjects (age and gender matched) and their parents were simultaneously selected from the same geographic area as control. Their gene polymorphism of MTHFD G1958A locus was examined with PCR-RFLP, and serum folic acid and homocysteine (Hcy) levels were tested with radio-immunoassay and fluorescence polarization immunoassay (FPIA). Results There existed gene polymorphism at MTHFD G1958A locus in healthy subjects living in North China. The percentages of GG, GA, and AA genotype were 57.98%, 35.57%, and 6.45% respectively, and the A allele frequency was 24.23%, which was significantly different from Western population. No difference was observed when comparing genotype distribution and allele frequency between the case and control groups, so was the result from the comparison between genders. The A allele frequency of arterial septal defect patients‘ mothers (10.87%) was significantly lower than that of controls (28.15%) (P=0.014), with OR=0.31 (95% CI: 0.09-0.84), and no difference in the other subgroups.The percentage of at least one parent carrying A allele in arterial septal defect subgroup (43.48%) was significantly lower than that in controls (69,64%) (P=0.017), with OR=0,34 (95% CI: 0.12-0.92). The analysis of genetic transmission indicated that there was no transmission disequillibrium in CHD nuclear families. Their serum folic acid level was significantly higher than that of controls (P=0.000), and Hcy level of the former was higher than that of the latter with no statistical significance(P＞0.05). Serum Hcy and folic acid levels of mothers with gene mutation were lower than those of mothers with no mutation.Conclusion No significant difference of genotype distribution and allele frequency existed between CHD patients and healthy population. MTHFD G1958A mutation in parents (particularly in mother) can decrease the risk of arterial septal defect in offspring. The possible mechanism of protection might be mutation, which can increase MTHFD enzyme activity, folic acid metabolism and homocysteine remethylation, and decrease Hcy level.     

Relationship between polymorphism of cystathionine beta synthase gene and congenital heart disease in Chinese nuclear families

Objective To study the relationship between polymorphism of cystathionine beta synthase （CBS） gene and development of congenital heart disease （CHD）. Methods One hundred and twenty-seven CHD case-parent triads were recruited from Liaoning Province as patient group, and 129 healthy subjects without family history of birth defect were simultaneously recruited as control group together with their biological parents. For all subjects the polymorphism of CBS gene G919A locus was examined by PCR-ARMS method, Results The frequencies of three genotypes （w/w, w/m, and m/m） in control group were 27.2%, 58,4%, and 14.4%, respectively, with no significant difference in gender. A significant difference in the allele frequency was found between CHD patients and controls, the wild allele frequency was 67,9% in patients and 55.7% in controls CHD parents＇ genotype distribution was significantly different from that in controls. Further comparison of each type of CHD showed that genotype frequencies in several CHD subtypes were significantly different from those in their corresponding controls. The results of TDT analysis showed that no allele transmission disequilibrium existed in CHD nuclear families. Conclusions CBS gene G919A mutation is associated with the development of CHD, and the mutated allele may decrease the risk of CHD.     

河南汉族冠心病患者内皮型一氧化氮合酶基因多态性分析

目的:探讨河南汉族冠心病患者内皮型-氧化氮合酶(eNOS)基因第4内含子VNTR的多态性.方法:依据VNTR位点侧翼序列设计引物,采用PCR方法检测河南汉族正常对照组(n=240)和冠心病人群(n=207)基因型,并对2组基因型与等位基因频率进行对比.结果:正常对照组和冠心病人群eNOS基因VNTR均存在a、b2种等位基因以及aa、ab、bb 3种基因型,冠心病组发现1例罕见ac基因型,但等位基因与基因型的分布频率存在差异.冠心病人群携带a等位基因的频率(20.29%)及aa纯合子(3.87%)的基因型频率均高于正常对照组(P＜0.05).结论:eNOS基因的a等位基因与冠心病相关,携带a等位基因者具有易患冠心病的危险性.     

缺血性脑血管病患者蛋氨酸合成酶基因A2756G多态性检测

目的:检测缺血性脑血管病患者蛋氨酸合成酶(MS)基因A2756G多态性.方法:运用PCR-限制性片段长度多态性(RFLP)技术对512例缺血性脑血管病患者及500例健康对照者进行MS基因多态性检测.结果:缺血性脑血管病患者组MS A2756G A/A、A/G、G/G基因型频率分别为88.9%、10.7%和0.4%,A和G等位基因频率分别为94.2%和5.8%.对照组A/A、A/G、G/G基因型频率分别为84.6%、15.0%和0.4%,A和G等位基因频率分别为92.1%和7.9%.2组基因型及等位基因频率差异均无统计学意义(P＞0.05).结论:MS基因A2756G多态可能不足以构成河南汉族人群缺血性脑血管病发病中的一个独立的遗传危险因子.     

13q33LOC122330位点与精神分裂症的关系

目的 :在中国北方汉族精神分裂症患者和其健康父母组成的 1 0 5个核心家系中探讨1 3q33LOC1 2 2 330位点与精神分裂症的关系。方法 :从全血中提取基因组 DNA,应用 PCR- RFLP方法检测 LOC1 2 2 330位点编码序列第 864个碱基从异亮氨酸到蛋氨酸的错义突变单核苷酸多态性 ( rs942 35 8)的分布。结果 :1 rs942 35 8基因型频率的分布符合 Hardy- Weinberg平衡 ;2单倍型相对风险分析 ( HRR)表明 ,rs942 35 8与精神分裂症无关联 (χ2 =0 .61 9P>0 .0 5 ) ;3传递不平衡检验( TDT)显示 ,父母和受累子女之间不存在显著的传递不平衡 (χ2 =0 .2 2 2 ,P>0 .0 5 ) ,即杂合父母传递给受累子女的等位基因无差异 ;4等位基因频率与精神分裂症的各种临床症状不相关。结论 :LOC1 2 2 330位点 rs942 35 8可能与精神分裂症无关 ,但不能排除该位点其它 SNPs与精神分裂症有关联     

基因芯片技术对血管紧张素转换酶、血管紧张素原及内皮型一氧化氮合酶基因多态性与冠心病关系的研究

目的:评估血管紧张素转换酶(ACE)基因、血管紧张素原基因(AGT)及内皮型一氧化氮合酶基因(eNOS)多态性与冠心病的关系,并分析ACE、AGT和eNOS基因多态性在冠心病的发生和发展中是否存在协同作用。方法:选择冠心病患者133人及对照者154人,用基因芯片技术检测ACE、AGT和eNOS基因多态性,并对比其基因型及等位基因频率。结果:冠心病组ACE DD,AGT TT和eNOS TT基因型频率与对照组相比有显著性差异,分别为OR=2.17,P<0.01,OR=2.63,P<0.001,OR=8.50,P<0.05。ACE、AGT和eNOS基因多态性与冠心病明显相关。同时携带ACE DD和AGTTT基因型或AGT TT和eNOS TT基因型者与冠心病明显相关(OR=3.47,P<0.01,OR=1.05,P<0.01)。结论:ACE、AGT和eNOS基因多态性可能是中国人冠心病的危险因素。基因芯片技术为研究多种易感基因与冠心病的相关性提供了一项高效、敏感的方法。