TLRs多态性与宫颈癌易感性之间的关系

目的:探讨汉族人群TLRs基因多态性与宫颈癌易感性之间的关系。方法:选取102例宫颈癌患者及100例健康查体正常人作为研究对象,PCR直接鉴定TLR2(-196 to-174 del)基因型;PCR结合限制性片段长度多态性技术(PCR-RFLP)检测分析TLR3c.1377C/T,TLR4 Asp299Gly、Thr399Ile,TLR9 1486T/C及G2848A位点多态性;然后选取具有代表性的标本进行测序,对酶切结果进行验证。结果:宫颈癌组和健康对照组中均未发现TLR4 Asp299Gly和Thr399Ile两个位点存在单核苷酸多态性。病例组和对照组的TLR2(-196 to-174 del)、TLR3c.1377C/T、TLR9 1486T/C、G2848A的基因型和等位基因频率未见显著差异。结论:TLR2(-196 to-174 del),TLR3c.1377C/T,TLR4Asp299Gly、Thr399Ile,TLR9 1486T/C、G2848A多态性与宫颈癌易感性无明显关系。     

白介素-12B基因多态性与原因不明复发性流产的易感性研究

目的:探讨IL-12B基因启动子区多态性与山东潍坊地区汉族育龄期妇女原因不明复发性流产(URSA)的相关性。方法:采用直接测序法检测83例原因不明复发性流产孕妇和90例正常孕妇IL-12B基因的启动子区的多态位点,并分析IL-12B基因多态性与原因不明复发性流产的关系。结果:IL-12B的启动子区检测到2个单核苷酸多态性(SNPs)(-671G/T和-405G/T)。2个SNPs紧密连锁,多态性分布符合Hardy-Weinberg平衡。URSA组和对照组的基因型和等位基因频率分布比较,差异无统计学意义(P0.05)。单体型分析显示,-671T/-405G频率在URSA组和正常对照组分别为0.65和0.41,URSA组显著高于正常对照组,差异有统计学意义(P0.05)。结论:正常孕妇IL-12B基因启动子区存在多态性变异,其中-671G/T和-405G/T构成的单体型-671T/-405G可能与URSA的易感性有关。     

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。     

原因不明复发性流产患者细胞毒性T淋巴细胞抗原4基因第一外显子49位点A/G基因多态性研究

目的探讨细胞毒性T淋巴细胞抗原4（CTLA-4）基因第一外显子49位点A／G基因多态性与原因不明复发性流产（URSA）发病的相关性。方法采用PCR限制性片断长度多态性方法（PCR-RFLP）,检测168例URSA患者（URSA组）和117例有正常生育史的妇女（对照组）CTLA-4基因第一外显子49位点A／G多态性,并比较等位基因G／A、基因型AA／AG／GG、表型A＋（AA＋AG）／G＋（GG＋AG）分布频率的差异。结果URSA组等位基因G的出现频率为68．4％（230／336）,对照组为59．4％（139／234）,两组比较,差异有统计学意义（P〈0．05）;基因型GG的出现频率URSA组为48．8％（82／168）,对照组为33．3％（39／117）,两组比较,差异也有统计学意义（P〈0．05）;URSA组基因型AG、基因表型A＋（AA＋AG）的频率分别为39．3％（66／168）、51．2％（86／168）,对照组分别为52．1％（61／117）、66．7％（78／117）,两组比较,差异均有统计学意义（P〈0．05）。结论CTLA-4基因第一外显子49位点A／G多态性与URSA的发生存在相关性,并可能参与流产发生的免疫病理过程。     

肿瘤坏死因子α308G/A位点基因多态性与宫颈癌易感性的Meta分析

目的:系统评价肿瘤坏死因子α(TNF-α)308G/A位点基因多态性与宫颈癌发生风险的关系。方法:检索Pub Med、Embase、Cochrane Library、中国学术期刊全文数据库(1979—2014.1)、维普数据库(1989—2014.1)和万方医学数据库(2000—2014.1)有关TNF-α308G/A基因多态性与宫颈癌发病风险关系的文献,按照纳入、排除标准选择试验并评价质量,从中提取有效数据进行Meta分析。结果:最终纳入16篇病例对照研究。Meta分析结果显示,总人群的TNF-α308G/A中A vs.G基因型与宫颈癌易感性差异有统计学意义(OR=1.242,95%CI为1.044~1.478,P=0.014)。亚组分析显示,亚洲人各基因型均不能增加宫颈癌发病风险(均P0.05)。白种人中基因型A vs.G(OR=1.299,95%CI为1.052~1.603,P=0.015),AA vs.GG(OR=1.627,95%CI为1.044~2.534,P=0.031),AA vs.(GA+GG)(OR=1.616,95%CI为1.033~2.527,P=0.036),与宫颈癌发生差异有统计学意义。非洲人中基因型GA vs.GG(OR=1.533,95%CI为1.023~2.297,P=0.039),GA vs.(AA+GG)(OR=1.531,95%CI为1.024~2.291,P=0.038),与宫颈癌发生有关。通过Bgger′s及Egger′s检验判断发表偏倚,结果可见A vs.G(P分别为0.029及0.030)存在发表偏倚。结论:TNF-α308G/A基因可能增加宫颈癌发病风险,但仍需大样本的高质量研究进一步证实。     

钙蛋白酶10基因多态性与多囊卵巢综合征患者糖耐量及脂代谢异常的相关性

目的 探讨钙蛋白酶10(CAPN-10)基因56位点单核苷酸多态性(SNP-56)与多囊卵巢综合征(PCOS)患者糖耐量及脂代谢异常的相关性.方法 选取山东地区PCOS患者334例(PCOS组),健康妇女304例(对照组),采用熔解温度不同的基因分型法检测CAPN-10基因SNP-56,并采用免疫化学发光法测定泌乳素、卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇、睾酮水平.PCOS组同时测定血糖、血脂、血清胰岛素水平.结果 (1)基因型及等位基因频率分布两组比较,差异均无统计学意义(P＞0.05).(2)在PCOS组,口服葡萄糖耐量试验(OGTF)180 min血糖水平AA基因型者为(5.7±2.2)mmol/L,GA基因型者为(4.9±1.2)mmol/L,GG基因型者为(4.9±1.4)mmol/L,分别比较,差异均有统计学意义(P均＜0.01);总胆固醇(TC)水平AA基因型者为(4.9±1.0)mmol/L,GA基因型者为(4.5±0.9)mmol/L,两者比较,差异也有统计学意义(P＜0.05).(3)在PCOS组,有糖尿病家族史者共44例,AA基因型频率为22.7%(10/44),GA+GG基因型频率为77.3%(34/44),GG基因型频率为34.1%(15/44);无糖尿病家族史者共290例,AA基因型频率为11.0%(32/290),GA+GG基因型频率为89.0%(258/290),GG基因型频率为47.2%(137/290),有无糖尿病家族史者AA与GA+GG基因型频率比较及AA与GG基因型频率比较,差异均有统计学意义(x2=4.751,x2=5.697;P均＜0.05).在PCOS组,有肿瘤家族史者共21例,AA基因型频率为33.3%(7/21),GA+GG基因型频率为66.7%(14/21),GG基因型频率为19.0%(4/21);无肿瘤家族史者共313例,AA基因型频率为11.2%(35/313),GA+GG基因型频率为88.8%(278/313),GG基因型频率为47.3%(148/313).结论 (1)CAPN-10基因SNP-56与PCOS的遗传易感性无明显相关性,但与PCOS患者糖、脂代谢异常有明显相关性.(2)CAPN-10基因SNP-56与PCOS患者糖尿病家族史及肿瘤家族史相关,应重视对高危PCOS个体的随访.     

PDCD6单核苷酸基因多态性与子宫内膜异位症易感性的研究

目的:探讨程序性死亡细胞因子6(PDCD6)基因多态性与子宫内膜异位症(EMT)的遗传易感性的关系。方法:选择我院2013年12月至2014年5月收治的单纯卵巢子宫内膜异位囊肿患者25例及无血缘关系的健康育龄妇女25例为研究对象,利用聚合酶链反应(PCR)及飞行时间质谱分析法对PDCD6基因两个位点rs4957020和rs4957014进行分析。结果:rs4957020基因上携带T等位基因的基因型(CT+TT)可提高EMT发病的风险(OR=3.273,95%CI 1.008~10.621,P0.05),同时,rs4957014基因上携带G等位基因的基因型(GT+GG)可提高EMT发病的风险(OR=3.431,95%CI 1.026~11.476,P0.05)。结论:PDCD6基因可能是导致EMT发生、发展的一个新的易感基因。     

新疆维、汉妇女miR-146a(rs2910164)基因多态性分析

目的:探讨新疆维吾尔族与汉族妇女间miR-146a(rs2910164)基因多态性的差异,分析其与宫颈癌发生发展的关系。方法:PCR-RFLP法检测宫颈炎、CIN及宫颈癌组织中miR-146a(rs2910164)基因分型。结果:维吾尔族和汉族间的miR-146a(rs2910164)基因型构成比差异显著(χ2=57.927,P0.001),维吾尔族GG基因型比例显著高于汉族。Logistic回归分析发现,维吾尔族(OR=3.332,95%CI 1.411~7.868)和肿瘤直径≥4cm(OR=3.792,95%CI 1.162~12.376)与miR-146a(rs2910164)基因型GG/CG基因型显著相关。结论:新疆维吾尔族与汉组妇女间miR-146a(rs2910164)基因多态性差异显著,miR-146a(rs2910164)等位基因G可能参与宫颈癌的病情发展。     

人β防御素-2及Toll样受体基因多态性与早产胎膜早破的相关性研究

目的探讨人β防御素-2(β-defensin-2,HBD-2)及Toll样受体(TLR)基因多态性与早产胎膜早破的相关性。方法选取2015年5月至2017年5月济南市人民医院产科收治的105例早产胎膜早破患者为研究对象(早产胎膜早破组),并选择40例正常足月分娩产妇为对照组,采用聚合酶链反应-限制性内切酶片断长度多态性(PCR-RFLP)法检测两组胎膜组织中HBD-2、TLR受体单核苷酸多态性(SNP)。结果早产胎膜早破组HBD-2基因CC型占58.10%、CG型占28.57%、GG型占13.33%,对照组CC型占72.50%、CG型占22.50%、GG型占5.00%,两组差异显著(P0.05),Arg753Gln位点TLR2基因突变率大于对照组,但差异无统计学意义(7.62% vs 5.00%,P0.05),Asp299Gly/Thr399Ile位点TLR4基因突变率大于对照组,但差异无统计学意义(5.71% vs 0.00%,P0.05)。结论 HBD-2携带GG基因型、等位基因G过多分布女性可能是导致早产胎膜早破的易感人群,Arg753Gln位点TLR2、Asp299Gly/Thr399Ile位点TLR4基因多态性与早产胎膜早破无相关性。     

VEGF基因多态性与不明原因复发性自然流产发生风险相关性的meta分析

目的:评价血管内皮生长因子(VEGF)基因的多态位点与不明原因复发性自然流产发生风险的相关性。方法:检索Pubmed数据库、Medline数据库、Cochrane图书馆数据库、中国知网(CNKI)、维普中文科技期刊数据库、万方数据库、中国生物医学文献数据库中有关VEGF基因多态性与不明原因复发性自然流产的病例-对照研究,对纳入的研究进行质量评价,采用Rev Man5.3软件进行数据分析。结果:最终纳入11篇文献对VEGF基因的-634G/C(rs2010963)、+936C/T(rs3025039)、-2578C/A(rs699947)及-1154G/A(rs1570360)4个位点进行评价,累计病例组1945例,对照组2074例。Meta分析结果显示,在VEGF基因的-634G/C位点,基因型CC发生复发性自然流产的风险高于基因型GG[P=0.03,OR=1.29,95%CI(1.03,1.63)];携带等位基因C妇女的发病风险高于携带等位基因G[P=0.02,OR=1.14,95%CI(1.02,1.27)]。+936C/T位点的CT、TT基因型及携带T等位基因发生复发性自然流产的风险高于CC基因型及携带C等位基因的女性[CT vs CC基因型:P0.0001,OR=1.40,95%CI(1.18,1.65),TT vs CC基因型:P=0.02,OR=1.72,95%CI(1.11,2.66),T vs C等位基因:P0.00001,OR=1.52,95%CI(1.30,1.78)];两组的-1154G/A、-2578C/A各基因型比较,差异均无统计学意义(P0.05)。结论:VEGF基因-634G/C(rs2010963)、+936C/T(rs3025039)位点的单核苷酸多态性与不明原因的复发性流产发生可能相关。     

Foxp3基因多态性与原因不明复发性自然流产易感性的关系

目的 探讨转录因子Foxp3基因多态性与原因不明复发性自然流产(URSA)易感性的关系.方法 分别采用PCR-限制性片段长度多态性技术(针对Foxp3基因的rs3761548、rs2294021位点)和PCR-等位基因特异性扩增技术(针对rs2232365、rs5902434位点),检测146例URSA患者(URSA组...     

原因不明复发性流产患者血中亚甲基四氢叶酸还原酶基因C677T和A1298C位点突变的研究

目的　探讨 5 ,10 亚甲基四氢叶酸还原酶基因C6 77T和A12 98C位点突变与原因不明复发性流产 (unexplainedrecurrentspontaneousabortion ,URSA)易感因素的相关性。 方法　采用PCR-限制性片段长度多态性方法 ,检测 14 7例原因不明复发性流产患者 (URSA组 )和 82例有正常妊娠史的妇女 (对照组 )血中亚甲基四氢叶酸还原酶基因C6 77T和A12 98C位点突变。结果　 ( 1)C6 77T的 3种基因型在URSA组和对照组总体分布存在显著性差异 (P =0 0 12 ) ,其中URSA组 :基因型CC占 33 3% ,CT占 5 3 1% ,TT占 13 6 % ,对照组 :基因型CC占 5 2 4 % ,CT占 5 1 5 % ,TT占 6 1%。两组 6 77CC基因表达差异有显著性 (P =0 0 0 5 ) ,URSA组C和T等位基因分别为 4 0 1%、5 9 9% ,两组基因分布情况比较 ,差异有显著性 (P <0 0 0 5 ) ;( 2 )A12 98C的 3种基因型在URSA组和对照组中总体分布情况比较 ,差异无显著性 ,12 98AA/AC/CC基因型和A/C等位基因频率比较 ,差异无显著性 (P >0 0 0 5 ) ;( 3)C6 77T/A12 98C连锁基因分析显示 ,8种连锁基因型中 ,URSA组 6 77CC/ 12 98AA表达频率显著降低 ,而 6 77(CT TT) / 12 98CC仅在URSA组中表达。结论　URSA与亚甲基四氢叶酸还原酶基因C6 77T和A12 98C位点突变有关。     

复发性流产患者外周血T淋巴细胞亚群及CD4^＋辅助性T淋巴细胞亚型的变化

】     

复发性流产患者外周血T淋巴细胞亚群及CD_4~ 辅助性T淋巴细胞亚型的变化

目的　探讨不明原因的复发性流产 (URSA)与T淋巴细胞免疫功能的关系。方法　采用酶联免疫斑点 (ELISPOT)法对 30例URSA患者和 2 4例正常妇女 (对照 )的外周血T淋巴细胞亚群及CD 4 辅助性T淋巴细胞 (Th细胞 )亚型进行检测。结果　与对照相比 ,URSA患者外周血中CD 4 辅助性T淋巴细胞百分率增多 (分别为 45 %、6 0 % ,P <0 0 5 ) ,Th1细胞的百分率升高 (14%与 2 6 % ,P<0 0 1) ,Th2细胞的百分率降低 (2 0 %与 7% ,P <0 0 5 ) ,Th1/Th2细胞比值增大 (0 73与 3 80 ,P<0 0 1)。结论　URSA的发生可能与CD 4 辅助性T淋巴细胞的增多及Th1细胞介导的细胞免疫功能的增强有关。     

亚甲基四氢叶酸还原酶基因多态性与不明原因的重复性自然流产的相关性研究

目的 :探讨同型半胱氨酸代谢酶基因亚甲基四氢叶酸还原酶 (MTHFR)基因多态性C6 77T、A12 98C及其联合基因型与不明原因重复性流产 (URSA)的关系。方法 :运用聚合酶链反应 -限制性片段长度多态性技术检测MTHFRC6 77T、A12 98C基因多态性。结果 :①患者组C/C基因型频率显著低于正常对照组者 ,总的突变T等位基因频率显著高于对照组者。MTHFRC6 77T基因型分布与不同年龄、地区、流产时间、流产性质无关 ,与流产次数显著相关。URSA组MTHFRA12 98C三种基因型频率与对照组相比差异无显著性 ,与患者不同临床特征无明显关联。A12 98C杂合子联合C6 77T杂合子基因型发生URSA的危险性无显著增高。结论 :MTHFRC6 77T基因多态性是URSA发病的遗传风险因素 ,而A12 98C基因多态性不是URSA发病的危险因素。     

Significance of Toll-like Receptor 4 Signaling in Peripheral Blood Monocytes of Pre-eclamptic Patients

Objective: Preeclampsia (PE) is a serious condition affecting pregnant women and placing both the mother and fetus at risk. While little is known about the pathogenesis of PE, there is evidence for involvement from the maternal innate immune system, specifically, signaling arising from monocytes. The present study was to explore the role of Toll-like receptor (TLR) 4 on peripheral blood monocyte in the pathogenesis of PE. Methods: This study included 22 patients with established preeclampsia and 23 healthy pregnant women (HP). All participants gave informed written consent for 4?mL of fresh venous blood to be collected into a tube containing heparin. The expression of TLR4 on monocytes was evaluated by flow cytometry (FCM). Monocytes were stimulated with LPS for 18?h and cytokine secretion (IL-6, IL-12P70, IL-10, and TNF-α) in supernatants was analyzed with Luminex platform (Luminex Corporation, Austin, TX). The expression of TLR4 and cytokine secretion (IL-6, IL-12P70, IL-10, and TNF-α) was compared between women with PE and healthy pregnant women. Results: Compared with controls, the percentage of TLR4+ monocytes was significantly higher in PE patients. Collected monocytes stimulated with lipopolysaccharide (LPS) to induce inflammation had increased cytokine production, and monocytes from PE patients produced more IL-6 and TNF-α, and less IL-10 than cells from healthy participants. PE patients also showed a positive correlation between the percentage of TLR4+ monocytes and serum levels of IL-6 and TNFα. Conclusions: These results suggest that TLR4 signaling may play a role in the pathogenesis of preeclampsia.     

Dendritic cells in the circulation of women with preeclampsia demonstrate a pro-inflammatory bias secondary to dysregulation of TLR receptors

Toll-like receptors (TLRs) are central components of the innate immune system that recognize both microbial ligands and host products released during tissue damage. Data from epidemiologic studies and animal models suggest that inappropriate activation of the immune system plays a critical role in the development of preeclampsia. This study evaluates in a systematic fashion the expression and function of TLRs in the circulation of patients with preeclampsia compared to healthy pregnant controls. We evaluated TLR expression and function in primary dendritic cells (DCs) of 30 patients with preeclampsia and 30 gestational age-matched healthy pregnant controls. DCs were stimulated with the different TLR ligands engaging TLR1/2, TLR2/6, TLR3, TLR4, TLR5, TLR7, TLR8 and TLR9. The expression of TLR-induced production of TNF-α, IFN-α, IL-6, and IL-12 were measured by multicolor flow cytometry. Basal expression of TLR3, TLR4 and TLR9 was significantly increased in DCs isolated from women with preeclampsia. Preeclamptic DCs also expressed significantly higher basal levels of cytokines. In contrast, preeclamptic DCs demonstrated a less robust response to stimulation with various TLR ligands as compared with healthy pregnant controls. Under basal conditions, DCs from preeclamptic individuals express higher levels of select TLRs and produce more pro-inflammatory cytokines as compared with healthy controls. As such, the ability of these cells to mount an inflammatory reaction in response to a TLR ligand is limited. These data demonstrate a dysregulated pattern of TLR expression and cytokine production in DCs from PE patients that may limit further activation by TLR engagement.     

Association between a functional single nucleotide polymorphism in the MDM2 gene and sporadic endometrial cancer risk

OBJECTIVES: MDM2 is an important negative regulator of the p53 tumor suppressor protein. A naturally occurring T/G single nucleotide polymorphism (SNP) in the MDM2 gene promoter, SNP309, causes an increase in MDM2 protein levels and impairment of p53 tumor suppressor activity. SNP309 occurs at a relatively high frequency in the general population and has been associated with accelerated tumorigenesis in hereditary Li-Fraumeni associated cancers as well as in sporadic soft tissue sarcomas. The objective of this study was to examine the association between SNP309 and sporadic endometrial cancer risk. METHODS: Genomic DNA was isolated from 73 patients with endometrial cancer and 79 healthy, female controls. The MDM2 gene promoter region was amplified by PCR and the SNP309 genotype determined by restriction enzyme digestion of the amplified DNA fragment. Unconditional logistic regression analysis was used to determine the relationship between genotypes and endometrial cancer risk and histopathologic features. RESULTS: The homozygous G/G genotype was found in 25% of endometrial cancer cases and 11% of controls. In an age-adjusted analysis of cases and controls, the G/G genotype increased the risk of endometrial cancer 2.76-fold (95% CI: 1.06, 7.20; p=0.03) compared to presence of a wild-type T allele (T/G and T/T genotypes). No association was found between the SNP309 G/G genotype and either endometrial cancer histology, grade, stage, or age at diagnosis. CONCLUSIONS: The MDM2 SNP309 homozygous G/G genotype may be a genetic variant that influences sporadic endometrial cancer susceptibility.