Parkin基因新的多态位点IVS3-20 T→C增加早发性帕金森病的发病风险

目的 确定parkin基因第3内含子区新的多态位点IVS3-20 T→C多态与帕金森病(Parkinson’s disease，PD)的相关性。尤其是该多态与PD发病年龄的关系。方法 经PCR扩增后。用变性高效液相色谱和DNA自动测序等方法分析了312例PD患者和236名正常对照parkin基因IVS3—20 T→C多态性位点分布频率的差异。结果 总体分析未发现C／C纯合型。PD组T／C基因型频率和C等位基因频率与对照组相比有明显升高，但差异无显著性。将PD组按年龄分层后。45岁以下PD患者IVS3—20 T→C多态T／C基因型频率(7．07%)明显高于正常对照组(2．12%)。OR=3．52，95%CI为0．97～13．13(P=0．0263)。C等位基因频率(3．90%)也明显高于正常对照组(1．06％)。OR=3．42。95%CI为0．96～12．57(P=0．0276)．而45岁以上PD组与正常对照组相比差异无显著性。年龄分层后的趋势分析显示parkin基因IVS3-20 T／C多态性相对危险度与PD发病年龄间存在负相关联系．结论 发现了parkin基因IVS3-20 T→C多态位点，并提示IVS3-20 T→C多态位点可能是中国人散发性早发PD的一个危险因素。     

单胺氧化酶B和NAD(P)H醌氧化还原酶基因多态性与帕金森病的关系

目的 探讨参与多巴胺代谢的单胺氧化酶B（monoamine oxidase B,MAO-B)内含子13A／G多态性,NAD（P）H醌氧化还原酶基因[NAD(P)H:quinone oxidoreductase,NQO1]cDNA609C/T多态性与帕金森病（Parkinson's disease,PD)遗传易感性的关系。方法 应用等位基因特异PCR扩增分析MAO－B基因的多态性,用PCR－RFLP分析NQO1基因多态性。结果 MAO－BA／G等位基因频率、各基因型频率PD组与对照组差异无显著性。NQO1基因T等位基因频率PD组（52％）高于正常对照组（43％）带T碱基的NQO1基因型频率PD组显著高于正常对照组（P＜0．05）,其患PD的相对危险度（odds ratio,OR)为3．8。在带有A等位基因MAO－B基因型的个体,带有T碱基因的T等位基因是PD的危险因素。MAO－B基因的A等位基因型与NQO1基因的T等位基因的基因型可相互协同,增加PD发生的风险。     

N-乙酰基转移酶基因多态性与帕金森病相关性研究

目的　探讨N -乙酰基转移酶 (NAT2 )基因多态性与散发性帕金森病 (Parkinson’sdisease ,PD)的关系。方法　应用自动实时荧光Light-Cycler技术 ,分析 88例PD患者和 112例健康人NAT2 4个位点的基因多态性 ,比较PD患者与对照组间频率差异。结果　早发PD组NAT2 6A等位基因频率与对照组比较有显著性差异 (P <0 .0 5 ) ,使患PD的危险度提高了 2 .0 8倍 (P <0 .0 5 ) ,NAT2 5A和NAT2 7A/B等位基因频率与对照组比较无显著性差异 (P >0 .0 5 ) ;晚发PD组NAT2 4个多态位点各等位基因频率与对照组比较无显著性差异 (P >0 .0 5 ) ;未检测到NAT2 14A等位基因。结论　NAT2 6A等位基因可能主要与早发PD的易感性相关 ,并参与了神经毒素的解毒。     

Parkin基因多态性与四川地区散发性帕金森病的相关性研究

目的了解四川地区散发性帕金森病parkin基因多态性与帕金森病(Parkinson’s disease,PD)的相关性。方法采用病例-对照研究,应用聚合酶链反应、限制性片段长度多态性、变性高效液相色谱及DNA测序等技术,对四川地区汉族人群198例帕金森病患者和187名正常对照parkin基因-258T/G及IVS3-20T/C多态性位点进行研究,并通过比较各多态性在PD患者和正常对照人群中的频率分布,探讨基因多态性与PD的可能关系。结果parkin基因-258T/G多态性PD组G等位基因频率明显高于对照组(52.5%vs43.3%),两组差异有统计学意义(χ2=6.17,P<0.025,OR=1.45,95%CI:1.04~1.86);进一步将PD组按发病年龄分层发现,只有50岁以上发病患者与对照组相比G等位基因频率差异具有统计学意义(χ2=9.048,P<0.01,OR=1.57,95%CI:1.08~2.06);同样,与对照组(69.51%)相比,PD组TG GG基因型频率(78.79%)亦增加,两者间差异具有统计学意义(χ2=3.854,P<0.05,OR=1.63,95%CI0.88~2.38);PD组不同基因型组间发病年龄差异具有统计学意义(P<0.05),GG基因型组的平均发病年龄比TT或TG基因型的发病晚近5年左右。parkin基因IVS3-20T/C多态性以CC基因型多见,TC型杂合子的频率较低,该实验未发现TT基因型。结论核心启动子区-258T/G多态G等位基因可能增加了四川地区汉族人群散发性PD发生的风险,尤其是50岁以后的发病风险;四川地区汉族人群IVS3-20T/C位点,以CC基因型多见,TC型杂合子的频率较低,无TT基因型。     

MTHFRC677T基因多态性与出生缺陷的相关性研究

目的探讨亚甲基四氢叶酸还原酶(MTHFR)C677T位点单核苷酸多态性与出生缺陷的相关性。方法回顾性的分析97例有不良孕产史(病例组)和210例无不良孕产史育龄妇女(对照组)MTHFR C677T的基因型,用统计学方法分析该位点单核苷酸多态性与胎儿出生缺陷的关系。结果病例组MTHFR C677T位点TT基因型频率为28.9%,高于对照组(18.5%),T等位基因频率为53.6%,同样高于对照组(42.8%),两组人群中该位点基因型、等位基因分布情况差异具有统计学意义(P0.05)。病例组中B1组、B3组MTHFR C677T位点TT基因型频率及T等位基因频率均高于对照组,且差异具有统计学意义(P0.05);而病例组中B2组、B4组MTHFR C677T位点TT基因型频率及T等位基因频率与对照组比较,差异无统计学意义(P0.05)。结论 MTHFRC677T位点单核苷酸多态性与神经管缺陷及染色体疾病的发生具有一定相关性,与先天性心脏病及唇腭裂的发生无明显相关性。     

徐州地区MTHFR基因C677T多态性与男性不育的相关性研究

目的探讨徐州地区亚甲基四氢叶酸还原酶(MTHFR)基因C677T多态性与男性不育的相关性。方法选取确诊的男性不育症患者715例做为本研究实验组,以有正常生育史且精液检测正常的男性572例做为对照组,根据MTHFR C677T基因多态性的结果,分析此多态性位点在两组中分布的差异。结果 MTHFRC677T位点等位基因T频率在不育症中显著高于健康对照组(P0.05)。MTHFRC677T位点CC基因型频率在不育症组中显著低于健康对照组,TT基因型频率在不育症中显著高于健康对照组(P0.05),CT基因型的频率在两组之间的差异无统计学意义(P0.05)。结论 MTHFR基因C677T单核苷酸多态性在徐州地区男性不育患者和健康人中,分布有显著差异,MTHFR基因C677T多态性与徐州地区男性不育有相关性。     

APOA5基因单核苷酸多态性与冠心病相关性研究

目的　探讨APOA5基因多态性与冠状动脉粥样硬化性心脏病 (coronaryheartdisease ,CHD)的相关性、与血脂关系及其在中国汉族人群中的分布。方法　用聚合酶链反应 限制性片段长度多态性分析APOA5与APOA4交界区域的T/C单核苷酸多态。结果　T/C单核苷酸多态位点等位基因T、C频率在CHD组和正常对照组分别为 0 .43 5、0 .5 65和 0 .3 74、0 .62 6。等位基因频率和基因型频率分布均符合Hardy Weinberg平衡定律。T/C基因多态性基因型频率 ,等位基因T、C频率在两组间差异有显著性(P <0 .0 5 ) ,且基因型CC的冠心病患者其血浆高密度脂蛋白水平显著高于其他基因型患者 (P <0 .0 1)。中国人T/C单核苷酸多态位点T、C等位基因频率与欧洲白人比较 ,差异存在非常显著性 ( 0 .3 74vs 0 .663、0 62 6vs 0 .3 3 7;P <0 .0 0 1)。结论　T/C单核苷酸多态性与CHD存在相关性 (P <0 .0 5 ) ,患者组CC基因型与血浆高密度脂蛋白水平密切相关 (P <0 .0 1)。     

食管癌发病风险与NAD(P)H:醌氧化还原酶1C609T基因多态性

目的 研究NAD(P)H：醌氧化还原酶l[NAD(P)H：quinone oxidoreductase l，NQO1]C609T基因多态性与食管鳞状上皮癌(esophageal squamous cell carcinoma，ESCC)发病风险的关系。方法 应用聚合酶链反应-限制性片段长度多态性方法检测193例ESCC患者及141名正常对照的NQOl C609T多态性位点的基因型。结果 ESCC患者的突变型(T)等位基因频率明显高于健康对照组(X^2=4．86，P=0．028)。ESCC患者的NQO1C／C和C／T基因型频率与健康对照组相比差异无显著性(X^2值分别为2．27和0．127；P值分别为0．132和0．721)，而ESCC患者的T／T基因型频率明显高于对照组(X^2=4．39，P=0．036)。与NQOlC／C及C／T基因型相比，T／T基因型可明显增加患ESCC的风险性(校正OR=1．8l，95％CI：1．04～3．15)，且在有上消化道肿瘤家族史的患者中尤为明显(校正OR=2．22，95％CI：1．18～4．17)。结论 对NQO1 C609T多态性位点的基因型检测可能对判断ESCC高危个体具有指导意义。     

TNF-α基因多态性与妊娠高血压的相关性研究

目的　探讨肿瘤坏死因子-α(tum or necrosis factor- alpha,TNF-α)基因启动子- 30 8G>A、- 85 0 C>T多态性与妊娠期高血压疾病的相关性。方法　应用聚合酶链反应-限制性片段长度多态性技术检测10 6例患者和10 8名健康孕妇的TNF- α基因启动子- 30 8G>A、- 85 0 C>T多态性。对两组之间的基因型频率和等位基因频率进行比较。结果　TNF-α基因启动子- 30 8位点TNF2等位基因频率和TNF2 / 1基因型频率在病例组明显升高(P<0 .0 5 )。- 85 0位点T等位基因频率和CT+TT基因型频率在对照组明显升高,差异有统计学意义(P<0 .0 5 )。由这两个多态性位点组成的不同基因型中,病例组TNF2 / 1CC基因型频率明显高于对照组,差异有统计学意义(P<0 .0 5 ) ,而TNF1/ 1TT基因型频率在对照组明显增高,差异有统计学意义(P<0 .0 5 )。结论　TNF- α- 30 8、- 85 0位点多态性与妊娠期高血压疾病相关,其中TNF- α基因- 30 8位点的突变是危险因素,- 85 0位点的突变可能是保护性因素。TNF2 / 1CC基因型可能是妊娠期高血压疾病的易感基因型。     

维生素D受体基因ApaⅠ和TaqⅠ位点多态性与帕金森病的相关性研究

目的 探讨维生素D受体(vitamin D receptor,VDR)基因ApaⅠ和Taq Ⅰ位点多态性与帕金森病(Parkinson's disease,PD)遗传易感性的相关性.方法 采用聚合酶链反应-限制性片段长度多态性技术和基因测序方法,检测285例中国北方汉族散发PD患者与285名正常对照VDR基因ApaⅠ和TaqⅠ位点多态性,并比较两组基因型和等位基因频率的差异.结果 ApaⅠ和Taq Ⅰ位点基因型和等位基因频率在PD组和对照组之间差异均无统计学意义(P＞0.05).将样本按性别及发病年龄分组后比较,ApaⅠ位点各亚组间基因型频率和等位基因频率差异亦无统计学意义(P＞0.05),而TaqⅠ位点的基因型分布在男性PD组(168例)与男性对照组(1 60名)之间差异有统计学意义(x2=4.187,P=0.032,OR=2.149,95％CI:1.011～4.567),男性PD组T等位基因频率较男性对照组显著增高(x2=3.867,P=0.036,OR=2.064,95％CI:0.989～4.307).结论 VDR基因ApaⅠ位点多态性与PD风险间无相关性,但TaqⅠ可能是男性PD的风险因素.     

单胺氧化酶A基因Fnu4HⅠ酶切位点多态性与帕金森病相关性研究

目的 探讨多巴胺代谢酶—单胺氧化酶A(monoamine oxidase A,MAO—A)基因Fnu4H Ⅰ酶切位点多态性与帕金森病(Parkinson‘s disease,PD)遗传易感性的关系。方法 应用聚合酶链反应—限制性片段长度多态性技术分析了MAO—A基因Fnu4H Ⅰ酶切位点多态性在PD患者与正常对照之间分布频率的差异。结果 PD组和对照组的G等位基因的频率分别为0．613和0．527(P＝0．039),TT基因型频率分别为0．303和0．415(P＝0．014),GG基因型频率分别为0．564和0．451(P＝0．021)。根据发病年龄分组后,表明这种差异主要存在于早发性PD和对照组之间,G等位基因在早发PD组和对照组中的频率分别为0．642和0．527(P＝0．029);GG基因型频率分别为0．585和0．451(P＝0．032);按性别分组后发现差异仅存在于男性PD和对照组之间,G等位基因的频率在男性PD组和对照组中分别为0．669和0．500(P＝0.005)。结论 MAO—A基因Fnu4H Ⅰ位点在对照组和PD患者之间的分布差异有显著性,G等位基因和GG基因型频率在PD组中较高,支持MAO—A基因多态性与PD相关的假说,而且与PD发病年龄和性别有关。     

Association analysis of NAD(P)H:quinone oxidoreductase gene 609 C/T polymorphism with Alzheimer's disease

Alterations of the NAD(P)H:quinone oxidoreductase (NQO1) activity are associated with Alzheimer's disease (AD). A polymorphism consisting of a single nucleotide (C-->T) change at position 609 of NQO1 influences the NQO1 activity. Therefore the NQO1 C609T polymorphism may confer susceptibility for AD developing. To test the hypothesis, we have performed an association study between the NQO1 gene polymorphism C609T and late-onset Alzheimer's disease (LOAD) in Chinese population. Totally 104 LOAD patients and 128 controls were enrolled in our data set. All subjects were genotyped for NQO1 and Apolipoprotein E (APOE). There were no significant differences in NQO1 genotype or allele frequencies between cases and controls. Likewise, with the stratification of APOE psilon4 status, no statistical difference was observed between cases and controls. Our findings suggested that this polymorphism might not represent additional genetic risk factor for LOAD. However, the present study cannot exclude NQO1 as a possible candidate for LOAD. Further study in a larger population and biological functional analysis of NQO1 gene is required to verify the role of NQO1 in LOAD.     

α-共核蛋白基因启动子区微卫星多态与晚发散发性帕金森病的关联研究

目的　探讨α 共核蛋白基因启动子区微卫星多态与晚发散发性帕金森病 (Parkinson’sdis ease ,PD)发病风险间的关系。方法　采用扩增片段长度多态方法和微卫星荧光标记 半自动基因分型技术 ,对上海汉族 13 5例晚发性散发性PD患者和 170名正常人进行α 共核蛋白基因微卫星多态分析 ,并通过比值比 (oddsratios ,OR)与PD进行相关分析。结果　病例 对照组间各等位基因频率分布差异有显著性( χ2 =14 .73 ,df =7,P =0 .0 4)。PD组 2 69bp等位基因频率最高 ( 0 .3 89) ,对照组中 2 71bp等位基因频率最高 ( 0 .3 5 9)。≤ 2 67bp等位基因与PD呈正关联 (OR =5 .2 2 8,95 %CI :1.2 48～ 2 7.2 0 2 ,χ2 =6.416,P =0 .0 11) ,而 2 73bp等位基因与PD呈负关联 (OR =0 .63 8,95 %CI :0 .44 0～ 0 .92 6,χ2 =5 .64 4,P =0 .0 18) ;病例 对照组间各基因型的分布差异无显著性 ( χ2 =16.3 68,df =12 ,P =0 .175 )。但含有≤ 2 67bp等位基因的基因型可增加PD的发病风险 (OR =4.5 94,95 %CI :0 .94～ 2 2 .49,χ2 =4.2 2 4,P =0 .0 4)。PD组杂合度为 40 % ,对照组为 5 0 %。结论　α 共核蛋白基因启动子区的微卫星多态与晚发性散发性PD的发病风险有关。     

The NAD(P)H:quinone oxidoreductase I C609T polymorphism modifies the risk of Barrett esophagus and esophageal adenocarcinoma.

PURPOSE: The role of genetic susceptibility to esophageal adenocarcinoma and its precursor lesion Barrett esophagus has not been fully elucidated. This study investigated the effect of polymorphisms in the manganese superoxide dismutase (MnSOD) and NAD(P)H:quinone oxidoreductase 1 (NQO1) genes in modulating the risk of developing Barrett esophagus or esophageal adenocarcinoma. METHODS: A total of 584 patients (146 esophagitis, 200 Barrett esophagus, 144 esophageal adenocarcinoma, and 94 controls) were genotyped for the MnSOD C14T and NQO1 C609T polymorphisms using polymerase chain reaction and restriction fragment length polymorphism analysis. RESULTS: The NQO1 TT genotype was less common in Barrett esophagus (2.0%) and esophageal adenocarcinoma (1.4%) patients, compared with both esophagitis patients (7.6%) and controls (5.4%). After adjustment for sex, age, body mass index, reflux symptoms, and smoking status, patients with the homozygous TT genotype had a 4.5-fold decreased risk of developing Barrett esophagus (odds ratio = 0.22, 95% confidence interval = 0.07-0.76, P = 0.01) and a 6.2-fold decreased risk of esophageal adenocarcinoma (odds ratio = 0.16, 95% confidence intervals = 0.03-0.94, P = 0.04) compared with individuals with the TC and CC genotypes. No significant differences between groups were observed for the MnSOD polymorphism (P = 0.289). CONCLUSIONS: Overall, the results of this study suggest that the NQO1 TT genotype may offer protection from reflux complications such as Barrett esophagus and esophageal adenocarcinoma.     

ATM基因rs227060位点单核苷酸多态性与肺癌易感性的相关性

目的:探讨共济失调毛细血管扩张症突变基因(ataxia telangiectasia mutated,ATM)rs227060位点单核苷酸多态性(single nucleotide polymorphisms,SNPs)与肺癌易感性之间的相关性.方法:采用聚合酶链反应-SNP敏感性分子开关方法,检测225例肺癌患者和128例健康体检者ATM基因rs227060多态位点等位基因以及基因型频率分布特点;并应用非条件Logistic回归法统计分析rs227060单核苷酸多态性与肺癌的相关性.结果:rs227060多态位点共检测出CC,CT,TT三种基因型和C,T两种等位基因,其在肺癌组与对照组的基因型分布频率为:CC基因型17.3％与29.7％、CT基因型61,4％与59.3％、TT基因型21.3％与11％,两组间基因型频率和等位基因频率分布差异均有统计学意义(P＜0.05).在对ATM rs227060基因型的多态性分析过程中发现:吸烟史在肺癌组与对照组相比差异无统计学意义(P＞0.05),而年龄、性别、肿瘤家族史在肺癌组与对照组相比差异均有统计学意义(P＜0.05);且以CC基因型作为对照,携带TT基因型的个体患肺癌的风险是携带CT基因型个体的3.49倍(OR=1.829;95％CI:1.045～3.199).结论:ATM基因rs227060位点单核苷酸多态性与肺癌易感性存在相关性,且携带TT基因型可增加肺癌的发病风险.     

建立单管双向荧光PCR方法检测NQO1基因609C/T多态性

目的 建立单管双向荧光PCR方法快速测定人NAD(P)H:醌氧化还原酶1[NAD(P)H:quinone oxidoreductase 1,NQO1]基因609C/T多态性.方法 以人NQO1基因中的609C/T位点,设计双向引物,优化反应条件,应用SYBR GreenⅠ双向荧光PCR扩增191份人基因组DNA标本,并通过对产物进行熔解曲线分析,根据产物Tm值进行等位基因单核苷酸多态性分型.对其中62份标本用经典的PCR-限制性片段长度多态方法进行基因型分型,验证结果准确性.结果 62份样本的单管双向荧光PCR方法的基因型结果与PCR-限制性片段长度多态法分型结果符合率100%.191份样本中,纯合野生型(CC)占28%,杂合型(CT)占50%,纯合突变型(TT)占22%.结论 单管双向荧光PCR方法检测NQO1基因609C/T多态性,操作简便、反应过程快速,结果直观,敏感性、准确性和稳定性好,适用于临床样本检测及流行病学调查研究.     

Vascular endothelial growth factor gene +813CC polymorphism of foetus is associated with preterm labour but not with pre-eclampsia in Turkish pregnant women

We investigated the effect of polymorphism at +813 locus of vascular endothelial growth factor (VEGF) gene on predisposition to preterm labour and pre-eclampsia (PE). We examined polymorphism of the VEGF +813 gene of foetuses from umbilical cord blood in 31 cases of preterm labour, 34 pre-eclamptic and 58 healthy term labour. VEGF +813 gene polymorphisms were studied using a polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. In preterm group, foetal CC genotype was found at 80.6%, and CT genotype was seen at 19.4%. No any TT genotype was detected in preterm group. CC genotype of VEGF 813 gene was significantly more frequent than CT genotype (P = 0.04). Foetuses with CC genotype VEGF+813 gene have an increased risk for preterm labour. C allele frequency was 90.3 and 76.7% in preterm and control groups, respectively. T allele frequency was 9.7 and 23.3% in preterm and control groups, respectively. C allele was significantly associated with preterm labour (P = 0.02). OR of C and T alleles for preterm labour was 2.8 (CI: 1.1-7.2). In PE group, foetal CC genotype of +813 locus was found in 67.6%, and CT genotype was seen in 29.4%. Only one TT genotype was detected in 2.9% of PE group. There was no association between PE and VEGF gene genotypes and alleles at +813 locus. These results suggest that foetal VEGF gene polymorphism of +813 CC seems to be highly associated with preterm labour, whereas in PE, foetal VEGF gene polymorphism at +813 locus is not related. Especially, C allele was significantly associated with preterm labour. Carriage of the +813C allele of the VEGF gene has been found 2.8 times increased susceptibility to the development of preterm labour in Turkish women and may be an independent risk factor for prematurity. There was no association between PE and VEGF gene genotypes and alleles at +813 locus. We suggest to search for foetal aetiologies or genetic susceptibility in preterm labour, whereas in PE, not foetal, but maternal susceptibility is to be investigated.