类DNA甲基转移酶蛋白3田DNMT3L)基因SNP rs207O565多态性与少精症的相关性

目的:研究类DNA甲基转移酶蛋白3 (DNMT3L)基因rs207O565多态性与少精症的相关性.方法:应用PCR-RFLP分析方法,在198名少精症患者(少精症组)和249名正常的男性个体(正常对照组)中,对.NMT3L基因rs207O565位点的基因频率和基因型频率分布进行调查.结果:在少精症和正常男性中rs207O565位点的等位基因频率分布存在差异,少精症组中等位基因A的频率显著高于正常对照组(20.7% vs 14.9%,P=0.03).结论:DNMT3L基因rs207O565位点的多态性与少精症相关,等位基因A可能增加少精症的易感性.     

胎盘儿茶酚氧位甲基转移酶基因多态性与子痫前期的关系

目的:探讨胎盘儿茶酚氧位甲基转移酶(COMT)基因rs4680、rs6269位点单核苷酸多态性(SNP)与子痫前期发病的关系。方法:选择2011年3月至2012年8月在南方医科大学附属深圳市妇幼保健院产科住院分娩的孕妇217例,其中子痫前期孕妇92例为子痫前期组,健康孕妇125例为对照组,收集两组孕妇产后胎盘组织,提取胎盘DNA,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)平台及MassARRAY-IPLEX技术对COMT基因rs4680及rs6269位点进行检测,分析其基因型及等位基因分布情况。结果:胎盘组织COMT基因rs4680位点基因型GG、GA、AA的基因型频率和G、A等位基因频率在子痫前期组与对照组之间的分布差异均无统计学意义(P0.05)。两组胎盘COMT基因rs6269位点GG、GA、AA 3种基因型频率差异有统计学意义(P=0.019);且G、A等位基因频率在子痫前期组和对照组分布差异有统计学意义(χ2=6.382,P=0.012),A/G OR=1.707(95%CI=1.125~2.590)。胎盘COMT rs4680与rs6269单核苷酸多态性与孕妇血压无明显相关性。结论:胎盘COMT rs6269单核苷酸多态性与子痫前期发生相关,携带A等位基因是发生子痫前期的危险因素;胎盘COMT rs4680单核苷酸多态性与子痫前期发生无关。     

褪黑素受体1B基因rs10830963位点多态性与妊娠期糖尿病遗传易感性的关系

目的 探讨褪黑素受体1B(melatonin receptor 1B,MTNR1B)基因rs10830963位点单核苷酸多态性与妊娠期糖尿病(gestational diabetes mellitus,GDM)遗传易感性的关系. 方法 选择来自浙江省衢州市妇幼保健院的GDM孕妇87例(GDM组)和同期正常妊娠妇女91例(对照组),应用PCR扩增和DNA测序技术确定所有研究对象MTNR1B基因rs10830963位点等位基因和基因型频率分布,并比较2组之间rs10830963位点等位基因和基因型频率的差异.此外,利用Logistic回归分析判断风险等位基因与空腹血糖和糖化血红蛋白(glycosylated hemoglobin A1c,HbA1c)的关系. 结果 GDM组rs10830963位点G等位基因频率和GG基因型频率均明显高于对照组,G等位基因携带者和GG基因型妇女发生GDM危险性分别是C等位基因和CC基因型的1.53倍(OR=1.53,95％ CI:1.005～2.324,P=0.047)和2.16倍(OR=2.16,95％ CI:1.052～4.434,P=0.034).利用Logistic回归校正年龄、孕前体重指数(body mass index,BMI)和糖尿病家族史等因素之后,GG基因型妇女发生GDM的危险性仍然显著增加(OR=2.07,95％ CI:1.048～4.372,P=0.022).多元线性回归分析显示风险等位基因G与空腹血糖(0.068 mmol/L,P=0.015)水平及HbA1c(0.073％,P=0.028)水平呈显著正相关. 结论 MTNR1B基因rsl0830963位点单核苷酸多态性和GDM遗传易感性有关.rs10830963位点G等位基因为GDM发生的遗传风险因子.     

CTLA4基因rs231775位点多态性与淮安地区汉族孕妇子痫前期遗传易感性相关性的研究

目的探讨CTLA4基因单核苷酸多态性位点rs231775与淮安地区汉族子痫前期发病风险的相关性。方法选择2008年6月至2014年4月在江苏省淮安市妇幼保健院产科住院分娩的子痫前期患者392例,选择同期正常妊娠孕妇317例作为对照组,采用基质辅助激光解吸附电离飞行时间质谱对CTLA4基因rs231775位点进行基因分型。结果子痫前期组CTLA4基因rs231775多态性位点GG、AG和AA基因型频率分别为53%,39%和8%,等位基因频率G,A分别为72%和28%;而在对照组中GG、AG和AA频率分别为45%,41%和14%,等位基因频率G,A分别为65%和35%。经卡方检验rs231775位点在两组的基因型和等位基因频率之间差异有统计学意义(P=0.025)。结论淮安地区的人群CTLA4基因rs231775位点的多态性与子痫前期有一定的关联。     

转录因子FOXP3基因多态性与不明原因复发性自然流产的易感性研究

目的:基于不明原因复发性自然流产(URSA)的免疫耐受学说,探讨转录因子FOXP3基因多态性与URSA的易感性。方法:采用等位基因特异性扩增-聚合酶链反应(PCR-SSP)法检测FOXP3基因rs2232365A/G和rs5902434del/ATT多态性在146例URSA患者(URSA组)和112例健康体检者(对照组)中的基因型分布。结果:①rs2232365A/G的3种基因型在URSA组和对照组的分布差异具有统计学意义(P<0.05),且携带G等位基因明显增加URSA的风险(P=0.01,OR=1.61)。②rs59024343种基因型在两组的分布差异无统计学意义(χ2=4.62,P=0.10),但等位基因del缺失型频率URSA组较对照组高(χ2=4.40,P=0.036)。③单倍体型分析:G/del单体型明显增加URSA的发病风险(P=0.02,OR=1.53),而A/ATT单体型则对URSA的发病具有保护作用(P=0.02,OR=0.63)。结论:转录因子FOXP3基因启动子区rs2232365和rs5902434多态性与URSA的遗传易感性有关,携带G和del等位基因明显增加URSA的发病风险。     

尾加压素Ⅱ基因多态性与妊娠期糖尿病遗传易感性的关系

目的探讨尾加压素Ⅱ基因rs228648位点单核苷酸多态性与妊娠期糖尿病（GDM）发病的关系。方法应用聚合酶链反应-限制性内切酶片段长度多态性（PCR—RFLP）技术;采用病例一对照研究的方法,对中国北方地区无血缘关系的70例GDM孕妇（GDM组）和70例正常妊娠妇女（对照组）的尾加压素Ⅱ基因rs228648位点（G—A）进行单核苷酸多态性分析。结果（1）两组孕妇尾加压素Ⅱ基因rs228648位点各基因型频率分布均符合Hardy-Weinberg平衡榆验,具有群体代表性。（2）GDM组孕妇尾加压素Ⅱ基因rs228648位点G等位基因频率为70．7％,对照组为57．9％,两组比较,差异有统计学意义（P〈0．05）;GDM组孕妇尾加压素Ⅱ基因rs228648位点A等位基因频率为29．3％,对照组为42．1％,两组比较,差异也有统计学意义（P〈0.05）。而两组孕妇的G／G基因型频率比较（分别为52．9％和41．4％）,差异无统计学意义（P〉0．05）。（3）尾加压素Ⅱ基因rs228648位点的A／A基因型频率与GDM组呈负相关关系,经多因素logistie同归分析显示,其OR值为0．312,OR值的95％可信区间为0．108～0．900（P=0．031）。结论尾加压素Ⅱ基因rs228648位点单核苷酸多态性可能在GDM遗传易感性中起重要作用。G等位基因可能与GDM发生有关,而尾加压素Ⅱ基因rs228648位点的A型纯合子可能是GDM的重要保护因素。     

一级亲属伴有代谢异常疾病的PCOS患者易感基因研究

目的:探讨基因单核苷酸多态性(SNP)与多囊卵巢综合征(PCOS)的相关性。方法:收集2018年1月至2019年12月就诊于新疆医科大学第一附属医院妇科的289例PCOS患者的临床资料。选取其中一级亲属患有代谢异常性疾病的35例PCOS患者和41例健康人群作为研究对象,采用Infinium Asian Screening BeadChip Array芯片检测全基因组SNP。通过KEGG富集分析和PPI分析确定易感基因和候选SNP位点,候选SNP位点进行群体遗传学分析。结果:芯片检测总差异SNP位点1141个(P≤0.001),生物信息学分析筛选出的易感基因和SNP位点为GNAO1(rs2587885),PIK3CD(rs6541017)和FGF13(rs527687、rs683357和rs7060413)。群体遗传学分析结果显示,rs527687位点A等位基因、rs683357位点G等位基因、rs6541017位点A等位基因、rs2587885位点C等位基因、rs7060413位点G等位基因在PCOS组中的频率显著低于对照组(P<0.05,OR<1且95%CI<1)。在共显性、显性、隐性遗传模型中rs527687、rs683357、rs6541017、rs2587885、rs7060413不同的基因型跟一级亲属有代谢性疾病的PCOS发病有相关性。结论:一级亲属患有代谢性疾病的PCOS患者rs2587885、rs6541017、rs527687、rs683357和rs7060413 SNP位点多态性与PCOS的发病风险具有相关性。     

子痫前期患者内皮素受体A基因G-231A多态性的研究

目的:探讨内皮素受体A(EDNRA)G-231A基因多态性与子痫前期有无关联。方法:用聚合酶链反应-限制性片段长度多态性分析法(PCR-RFLP),对成都地区220例子痫前期患者(其中轻度71例,重度149例)和270例健康对照组孕妇EDNRA基因G-231A多态性进行检测。结果:子痫前期组和正常孕妇组EDNRA基因G-231A位点A等位基因频率分别为35.9%、31.3%,两组之间等位基因频率分布无显著差异(P>0.05);重度子痫前期组与轻度子痫前期组比较,A等位基因频率分别为36.3%、35.2%,二者无显著差异(P>0.05)。此外,子痫前期组和正常孕妇组不同基因携带者收缩压和舒张压水平无统计学差异(P>0.05)。结论:本研究未发现EDNRA基因G-231A多态性与子痫前期发病有关联。     

IDE基因和TCF7L2基因多态性与妊娠期糖尿病的相关性研究

目的探讨TCF7L2基因rs10885410、rs7100927位点和IDE基因rs10882066、rs11187007、rs7078413位点单核苷酸多态性(SNP)与妊娠期糖尿病(GDM)遗传易感性的关系。方法选取2011年9月至2013年8月在中国医科大学附属盛京医院住院分娩的GDM孕妇178例为病例组,同期121例葡萄糖耐量正常的妊娠妇女作为对照组。提取受试者全基因组DNA,对TCF7L2基因rs10885410、rs7100927位点和IDE基因rs10882066、rs11187007、rs7078413位点进行基因分型,并进行相关分析。结果 TCF7L2基因rs10885410、rs7100927位点各基因型和等位基因频率分布,两组间差异无统计学意义(P0.05)。IDE基因的rs11187007位点,病例组AA、AG、GG 3种基因型频率分别为4.49%、56.18%、39.33%,对照组分别为10.74%、38.84%、50.42%,两组比较,P0.05。AG基因型GDM的发病风险与AA基因型相比,OR值为3.457(95%CI 1.342~8.909,P=0.010),经Logistic回归校正混杂因素后,OR值为3.403(95%CI 1.072~10.803,P=0.038)。结论 IDE基因的rs11187007位点多态性可能与GDM遗传易感性有关;AG基因型可能是其发生的危险性因素。     

胰岛素降解酶基因多态性与多囊卵巢综合征相关性的研究

目的:探讨胰岛素降解酶基因(IDE)多态性与多囊卵巢综合征(PCOS)发病的关系。方法:用聚合酶链反应-限制性片段多态性技术,检测315例PCOS患者(PCOS组)和327例健康妇女(对照组)的IDE基因多态性。结果:IDE基因rs1887922位点基因型分布及等位基因频率在PCOS组总体以及各亚组(胰岛素抵抗组及非胰岛素抵抗组),与健康对照组之间比较均无显著差异。而IDE基因rs2209972位点基因型分布虽然在P- COS组总体及非胰岛素抵抗亚组与健康对照组之间比较无显著差异;但PCOS组总体相应位点的C等位基因频率高于对照组,差异有显著性(P<0.05),PCOS胰岛素抵抗亚组相应位点的基因型频率及等位基因频率均高于对照组,差异有统计学意义(P<0.05)。rs2209972位点不同基因型PCOS患者空腹胰岛素水平及HOMA-IR存在显著差异(P<0.05)。结论:胰岛素降解酶基因多态性可能与PCOS患者胰岛素抵抗相关。     

Association between single-nucleotide polymorphisms of DNMT3L and infertility with azoospermia in Chinese men

The gene for DNA methyltransferase 3-like protein (DNMT3L) is essential for normal spermatogenesis and may be involved with spermatogenetic impairment and male infertility. To explore the possible association between the DNMT3L gene and male infertility, this study investigated allele, genotype and haplotype frequencies of three single nucleotide polymorphism (SNP) loci, rs2070565, rs2276248 and rs7354779, of DNMT3L in 233 infertile patients with azoospermia and 249 fertile controls from a population of Chinese men using polymerase chain reaction/restriction fragment length polymorphism. Results showed that the frequencies of allele A (20.6% versus 14.9%; P = 0.022) and the allele A carrier (GA + AA; 37.8% versus 28.1%; P = 0.027) in azoospermic patients were significantly higher than those in controls at the rs2070565 locus. The haplotype AAA frequency was significantly higher (18.1% versus 12.4%; P = 0.02) while the haplotype GAA frequency was significantly lower (53.2% versus 62.1%; P = 0.007) in infertile patients compared with fertile controls. These results indicated that SNP rs2070565, as well as haplotypes AAA and GAA, may be associated with male infertility and suggest that DNMT3L may contribute to azoospermia susceptibility in humans.     

Association of common SNP rs1136410 in PARP1 gene with the susceptibility to male infertility with oligospermia

Purpose

This study aims to explore possible associations between polymorphisms of common SNP rs1136410 and rS1805405 in PARP1 gene and male infertility with spermatogenesis impairment.

Methods

The polymorphic distributions of SNP rs1136410 and rS1805405 were investigated by polymerase chain reaction and restriction fragment length polymorphism analysis in a Chinese cohort including 371 infertile patients with idiopathic azoospermia or oligospermia and 231 controls.

Results

Significant differences in the frequencies of allele and genotype of SNP rs1136410 were observed between patients with oligospermia and controls. The allele C (46.3 % vs. 36.4 %, P = 0.003) and genotype CC (22.6 % vs. 13.4 %, P = 0.014) significantly increased, whereas genotype TT (30 % vs. 40.7 %, P = 0.021) significantly decreased in patients with oligospermia compared with controls at this SNP locus.

Conclusions

These results indicated that genotype CC of SNP rs1136410 may increase the risk of oligosoermia and genotype TT of rs1136410 may have some protective effect from oligospermia, suggesting that the polymorphism of SNP rs1136410 in PARP1 gene may modify the susceptibility to male infertility with oligospermia.     

DNMT1基因多态性和被动吸烟与宫颈癌的相关性

目的：研究甘肃地区汉族女性DNA甲基转移酶1（DNMT1）基因多态性和被动吸烟与宫颈癌的相关性。方法：采用病例对照研究方法,选取甘肃地区100例宫颈癌患者和100例健康对照者,应用聚合酶链反应和基因测序法检测DNMT1基因多态性,比较不同基因型、被动吸烟与宫颈癌之间的关系。结果：被动吸烟者发生宫颈癌的相对危险性是无被动吸烟者的2.705倍（P＜0.05）;宫颈癌组rs16999593位点C等位基因频率高于对照组（P＜0.05）。结论：DNMT1基因的单核苷酸多态性和被动吸烟增加了宫颈癌的发病风险。     

Susceptibility to male infertility: replication study in Japanese men looking for an association with four GWAS-derived loci identified in European men

Purpose

A previous genome-wide association study in European men identified four single nucleotide polymorphism (SNP) loci associated with male infertility. Our aim was to replicate, if possible, the association of these SNPs with Japanese male infertility.

Methods

We genotyped four SNPs (rs5911500, rs10246939, rs2059807, and rs11204546) in 517 Japanese patients with male infertility and 369 fertile controls using SNP-specific real-time polymerase chain reaction TaqMan assays. Subsequently, we divided patients with male infertility into azoospermia (n = 417) and oligospermia subgroups (n = 70).

Results

The four SNPs previously identified in European men showed no significant association with collective male infertility in our Japanese cohort. However, allele frequency analysis did indicate a significantly higher frequency of the rs11204546 C allele of the OR2W3 gene in the oligospermia subset of infertility patients compared with controls (p = 0.0037; odds ratio = 1.74; 95 % confidence interval, 1.21–2.53).

Conclusions

Although this study was somewhat limited by overall sample size, the OR2W3 gene polymorphism rs11204546 was significantly associated with oligospermia in Japanese men, suggesting that OR2W3 might be involved in genetic susceptibility to Japanese male infertility as well as in European males.

Electronic supplementary material

The online version of this article (doi:10.1007/s10815-015-0468-4) contains supplementary material, which is available to authorized users.     

Relation of locus 1p13 rs646776 polymorphism with the risk of preeclampsia

Objective: This study aimed to assess the relation of locus 1p13 rs646776 (T/C) polymorphism with preeclampsia in Egyptian women. Methods: The study included 100 healthy pregnant female subjects and 100 preeclampsia patients. The genotypes of the polymorphisms were assessed. Endothelin-1 level was determined in plasma. Results: The major T allele of the 1p13.3 genomic region rs646776 polymorphism had a higher frequency in preeclampsia patients. Carriers of C allele had significantly lower endothelin-1 levels, lower systolic and diastolic blood pressure, decreased proteinuria, and increased HDL-C in the patients. Conclusion: The rare C allele of rs646776 polymorphism in chromosomal locus 1p13.3 is associated with decreased risk of preeclampsia.     

Polymorphic variants of DNMT3A and the risk of endometriosis

Objective

Overexpression of DNA methyltransferase 3A (DNMT3A) and aberrant methylation of various genes in eutopic endometrium have been demonstrated in women with endometriosis. We aimed to study whether DNMT3A polymorphisms could be a genetic risk factor for endometriosis and endometriosis-related infertility.

Study design

We studied 5 SNPs (rs2289195, rs7590760, rs13401241, rs749131 and rs1550117) located in the DNMT3A gene in 357 women with endometriosis and 640 controls.

Results

We did not observe significant differences between genotype and allele frequencies of rs2289195, rs7590760, rs13401241, rs749131 and rs1550117 SNPs in women with endometriosis, endometriosis-related infertility, and controls. The lowest p values of the trend test were observed for DNMT3A rs1550117 in endometriosis and endometriosis-related infertility (p_trend = 0.049 and p_trend = 0.055, respectively).

Conclusions

Our results did not supply evidence for the contribution of SNPs located in DNMT3A to either endometriosis or endometriosis-related infertility.     

DNA methyltransferase 3A promoter polymorphism is associated with the risk of human spontaneous abortion after assisted reproduction techniques and natural conception

Purpose

The aim of this study was to explore the association of the DNA-methyltransferase (DNMT)-3A and DNMT3B promoter polymorphisms with the risk of human spontaneous abortion after assisted reproduction techniques (ARTs) and natural conception.

Methods

We collected tissues from women who underwent abortion procedures: (a) chorionic villus samples (CVS) and muscle samples (MS) from spontaneous abortions conceived by ART and natural cycle (study group), n?=?152; and (b) CVS and MS from normal early pregnancy and second trimester (control group), n?=?155. The single-nucleotide polymorphism (SNP) ?448A?>?G in the DNMT3A promoter region and ?149C/T polymorphism of DNMT3B were determined by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) and confirmed by sequencing.

Results

The allele frequency of ?448A among pregnancy loss group and control group was 34.2 % vs. 16.5 %, respectively. Compared with GG carriers, the DNMT3A ?448AA homozygotes had an about 16-fold increased risk of spontaneous abortion [odds ratio (OR)?=?16.130, 95 % confidence interval (CI), 3.665–70.984], and AG heterozygotes had an OR of 2.027 (95 % CI, 1.247–3.293). However, the distribution of ?448A?>?G in individuals derived from ART pregnancies was not statistically significantly compared with those derived from spontaneous pregnancies (P?=?0.661). For DNMT3B, we observed genotype frequencies of 100 % (TT) in the study group and the control group.

Conclusions

The DNMT3A ?448A?>?G polymorphism may be a novel functional SNP and contribute to its genetic susceptibility to spontaneous abortion in Chinese women, and ART may not affect the distribution of ?448A?>?G in pregnancy loss and normal pregnancy. The observed TT genotype of DMNT3B suggests that this is the predominant genotype of this population. The findings provide new insights into the etiology of human spontaneous abortion.

     

DNA甲基转移酶在胚胎停育绒毛组织中的表达差异及临床意义

目的：探讨胚胎停育绒毛组织中DNA甲基转移酶（DNMTs）4种亚型DNMT1、DNMT3A、DNMT3B与DNMT3L的mRNA及蛋白表达差异,并探讨其临床意义。方法：随机选取2013年1月-2014年6月在青岛市立医院妇产科门诊就诊的经B型超声（B超）证实为胚胎停育而行清宫术的40例患者为观察组,并选取同期正常早孕要求人工流产的40例患者为对照组。①采用实时荧光定量聚合酶链反应（qRT-PCR）方法检测2组患者绒毛组织中DNMT1、DNMT3A、DNMT3B与DNMT3L mRNA的表达量;②用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶连结法（SP法）及蛋白质印迹法（Western blot）检测DNMT1、DNMT3A、DNMT3B与DNMT3L蛋白在观察组和对照组患者绒毛组织中的表达部位及定量差异。结果：①qRT-PCR结果显示,2组患者绒毛中DNMT1、DNMT3A、DNMT3B和DNMT3L mRNA的表达量比较差异无统计学意义（P＞0.05）;②免疫组化结果显示,DNMT1、DNMT3A、DNMT3B与DNMT3L在2组绒毛滋养细胞的细胞核或细胞质呈不同程度的阳性染色,同时,半定量分析结果显示观察组的DNMT3A蛋白表达量低于对照组,差异有统计学意义（P＜0.05）;③Western blot分析结果显示,观察组患者绒毛中DNMT3A蛋白的相对表达量低于对照组,差异有统计学意义（P＜0.05）;④胚胎停育绒毛组织中DNMT3A蛋白的表达量与DNMT1、DNMT3B和DNMT3L蛋白表达量之间无明显关联（均P＞0.05）。结论：胚胎停育绒毛组织中,DNMT3A蛋白水平的低表达可能参与了胚胎停育的发病机制。