【作者】 陈志芳；

【导师】 张怡； 丁岩；

【作者基本信息】 中南大学 ， 临床医学， 2013， 博士

【摘要】 第一部分新疆乌鲁木齐市宫颈癌HPV分型流行病学研究目的：调查乌鲁木齐市妇女生殖道不同型别人乳头瘤病毒(HPV)的感染状况以及在人群中的分布。方法：采用整群分层的抽样方法对乌鲁木齐市沙依巴克区18～64岁的女性开展以人群为基础的宫颈癌筛查。签署知情同意书后,对合格对象进行流行病学问卷调查。采集受试者宫颈脱落细胞,进行液基细胞学检测、HPV分型基因芯片检测。液基细胞学(LCT)检测,LCT异常者进行阴道镜及宫颈组织病理活检。以病理结果作为诊断宫颈CIN的金标准。结果：1.该人群HPV-DNA阳性检出率为20.28%,高危型HPV阳性人群中以16、58、52和18型最多,2.高危型HPV阳性率在18～34、35～44、45～64年龄组的阳性率依次为18.07%、29.64%和7.64%(x2=105.02,P<0.05)。其中35～44岁、17～34和45～64岁三个年龄组的阳性率差异均具有显著性(χ2=19.43,P<0.05；χ2=22.62,P<0.01；χ2=66.17, P<0.01；)。3.HPV感染率随着宫颈病变的加重呈趋势性增加(x22=697.72,P<0.05；)4.外来务工人员、白领阶层、居民区人员和服务业从业妇女HPV感染率分别为24.67%,12.19%,12.74和35.11%。服务业妇女HPV感染率明显高于白领人员、居民区人员及外来务工人员(χ2=74.46,P<0.05和χ2=84.93,P<0.05；χ2=13.41,P<0.05)；而外来务工人员HPV感染率明显高于白领人员、居民区人员(x2=27.56,P<0.05和x2=29.88,P<0.05)。结论：乌鲁木齐市HPV高危型别以16、58、52和18型最多,中年妇女是HPV高感染年龄组,不同职业中,服务业人员HPV感染率高,应加强监控。第二部分新疆汉族、维吾尔族、哈萨克族生殖道HPV感染分型的研究目的：调查新疆维吾尔族、汉族、哈萨克族妇女生殖道不同型别人乳头瘤病毒(HPV)的感染状况以及在人群中的分布。方法：采用整群分层的抽样方法对新疆18～64岁的女性开展以人群为基础的宫颈癌筛查。签署知情同意书后,对合格对象进行流行病学问卷调查。采集受试者宫颈脱落细胞,进行液基细胞学检测、HPV分型基因芯片检测。液基细胞学(LCT)检测,LCT异常者进行阴道镜及宫颈组织病理活检。以病理结果作为诊断宫颈CIN的金标准。结果：1.结婚次数,文化程度,吸烟史,人工流产数,避孕套使用,性伴数,近5年内的性伴侣数以及人群来源等与维吾尔族HPV感染相关,四个变量(文化程度、性伴侣数、避孕套使用和人群来源)进入了多因素分析模型。维吾尔族、汉族、哈萨克族比较：2.维吾尔族HPV感染率最高,与汉族、哈萨克族比较有统计学意义。3、汉族高危型HPV阳性人群中以16、58、52、18和31型HPV阳性者最多,维吾尔族高危型HPV阳性人群中以16、58、31、52和68、39型HPV阳性者最多,哈萨克族高危型HPV阳性人群中以16、18、58、33型HPV阳性者最多。4、三个民族HPV感染率随着宫颈病变的加重呈趋势性增加,三个民族C INⅡ及c INⅢ发生率比较,有统计学差异,其中维吾尔族发生率最高,与其他两组比较,均有统计学意义。结论：文化程度和避孕套使用是HPV感染的保护因素,性伴侣数和人群来源是HPV感染的危险因素。维吾尔族HPV感染率最高,三个民族高危型HPV感染型别不同,三个民族HPV感染率随着宫颈病变的加重呈趋势性增加。第三部分维吾尔族宫颈癌HPV与Treg免疫逃避机理的研究目的：探讨维吾尔族宫颈癌患者外周血中CD4+CD25+CD1271o/-调节性T细胞及血清中TGF-β、IL-10及IFN-γ的表达及意义。检测维吾尔族宫颈病变不同时期以及对照组外周血CD4+CD25High Treg及其特异标志物Foxp3的表达,探讨CD4+CD25HighTreg和Foxp3在维吾尔族宫颈癌发病机制中的作用。探讨维吾尔族宫颈癌人乳头瘤病毒与CD4+CD25+CD127-调节性T细胞的相关性。方法：采用流式细胞术检测66例维吾尔族宫颈癌患者、35例宫颈CIN Ⅱ-Ⅲ患者、45例宫颈CIN Ⅰ患者和40例正常对照组外周血中T细胞亚群、CD4+CD25+CD1271o/-Treg含量及CD4+CD25HighTreg含量,采用ELISA法检测血清中TGF-β、IFN-γ和IL-10的表达水平。并比较术前术后宫颈癌患者各组指标的变化。采用逆转录-聚合酶链反应(RT-PCR)检测CD4+CD25+T细胞中Foxp3mRNA表达水平。采用HPV (human papilloma viruse HPV)核酸扩增分型检测试剂盒检测66例维吾尔族宫颈癌患者、35例宫颈CIN(宫颈上皮内瘤样病变cervical intraepithelial neoplasia CIN)Ⅱ-Ⅲ患者、45例宫颈CINⅠ患者和40例正常对照组HPV分型。结果：1.宫颈癌与宫颈CIN Ⅱ-Ⅲ患者外周血中CD4+CD25+CD1271o/-Treg占CD4+T淋巴细胞的百分比以及各组细胞因子的含量与宫颈CIN Ⅰ组及正常对照组比较均有统计学差异(P均<0.05)。2.宫颈癌及宫颈CIN组IFN-γ含量与CD4+CD25+CD1271o/-T reg含量呈负相关(r=-0.585, P=0.000); TGF-β、IL-10与CD4+CD25+CD127lo/-Treg含量呈正相关(r=0.690, P=0.000, r=0.827, P=0.000)。3.宫颈癌患者手术前后外周血中CD4+CD25+CD1271o/-Treg与TGF-β、IL-10及IFN-y含量的比较差异均有统计学意义(P<0.05)。4.CD4+CD25+T细胞中Foxp3mRNA表达水平,宫颈癌组与宫颈CIN I组及正常对照组比较均有统计学差异(P均<0.05)。5.宫颈癌组及宫颈CINII-III组CD4+CD25HighTreg细胞表达率与Foxp3mRNA呈显著正相关(r=0.853, P=0.000; r=0.586, P=0.000)。6. HPV与Treg显著正相关(r=0.560, P<0.05)。结论：维吾尔族CD4+CD25+CD1271o/-Treg及相关因子TGF-β、IL-10及IFN-y及Foxp3参与了维吾尔族宫颈癌Treg免疫逃避的过程。维吾尔族宫颈癌HPV与Treg密切相关。更多还原

【Abstract】 The first chapter Epidemic research of human papillomavirus infection genotypes of cervical cancer in Urumqi in XinJiangObjective:To investigate the human papillomavirus infection genotypes and distribution in Urumqi Female Population reproductive tract.Methods:A questionnaire survey was conducted among the women aged18-64years in Urumqi by randomizing clustered sampling under stratification. All of patients received liquid-based cytology test(LCT) and HPV types. The patients who had abnormal cytological diagnosis were put up digital colposcopic exam. We Used pathologic diagnosis as a gold standard to diagnose CIN.Results:1.The positive rate of HPV was20.28%overall.2.The most prevalent genotypes of high-risk human papillomavirus (HR-HPV) were HPV16,58,52,18. The positive rate of HPV at different age groups, aged18-34,35-44,45-64years were18.07%,29.64%,7.64%, respectivly(P<0.05).3.The risk of cervical lesion significantly increased in women showing positive in cytology and HPV test. The HPV infection rate in different Occupations were different with the highest in service occupation(35.11%) while the lower appeared in works, white-collars and general residents. The HPV infection rate in works was significantly higher than that in general residents and white-collars. Conclusion:The most prevalent genotypes of high-risk human papillomavirus (HR-HPV) were HPV16,58,52,18in Urumqi. People aged35-44years are the most frequent infected groups in Urumqi. The HPV infection rate was highest among women having service related jobs and they should be closely monitored. The second chapter The investigation of reproductive tract human papillomavirus infection genotypes and distribution in Hans, Uygurs and Hazakhs of XinJiangObjective:To investigate the human papillomavirus infection genotypes and distribution in Hans, Uygurs and Hazakhs of XinJiang.Methods:A questionnaire survey was conducted among the women aged18-64years in Xinjiang by randomizing clustered sampling under stratification. All of patients received liquid-based cytology test(LCT) and HPV types. The patients who had abnormal cytological diagnosis were put up digital colposcopic exam. We Used pathologic diagnosis as a gold standard to diagnose CIN. Results:1. Among the total,the frequency of marriage, degree of culture, smoking history, the number of induced abortion, condoms using, the NO.of sexual partner, the NO.of sexual partner in recent five years and source of population had the relevant with human papilloma-virus infection of the Uygurs. Four variables(degree of culture,the NO.of sexual partner, condoms using and source of population)had significant in the multi-factor model.2.The positive rate of HPV of Uygurs was highest.,the comparison with Hans, Hazakhs had statistical significance.3.The most prevalent genotypes of high-risk human papillomavirus (HR-HPV) of Hans were HPV16,58,52,18,31. Uygurs’ were HPV16,58,31,52,68,39. Hazakhs’ were HPV16,18,58,33.4. The risk of cervical lesion significantly increased in women showing positive in cytology and HPV test. The comparison of incidence of C IN Ⅱ and C INⅢ of three nations had statistical significance. Uygurs’ were the highest.Conclusion:Degree of culture and condoms using were the protect factors, the NO.of sexual partner and source of population were the risk factors. The positive rate of HPV of Uygurs was highest.The genotypes of high-risk human papillomavirus (HR-HPV) of three nations were different. The risk of cervical lesion significantly increased in women showing positive in cytology and HPV test in three nations. The third chapter Research of mechanism of immune evasion in human papilloma viruse and CD4+CD25+CD127-Treg in cervical cancer of UygursObjective:To evaluate the expressions of CD4+CD25+CD127lo/-T cells in peripheral blood and TGF-β、IL-10and IFN-γ in serum from patients of Uygurs with cervical cancer. Through detect the level of CD4+CD25High T cell and the Foxp3in peripheral blood of cervical cancer patients of Uygurs,to investigate the effect of cervical cancer pathogenesis of CD4+CD25High T cell and the Foxp3of Uygurs. To investigate the correlation between human papilloma viruse(HPV) and CD4+CD25+CD127-T cells (Treg) of Uygurs.Methods:66patients with cervical cancer,35patients with CINII-Ⅲ,45patients with CINI and40normal controls were included in this study. The percentage of CD4+CD25+CD127lo/-T cells、CD4+CD25HighT population in CD4+T cells and T cells were evaluated by flow cytometric analysis. TGF-β、IL-10and IFN-γ were measured by ELISA.To compare the change of targets of preoperative and posto- perative of cervical cancer patients. The expression of Foxp3was evaluated by reverse transcription PCR. Typing of HPV (human papil-loma viruse) was evaluated by Nucleic acid amplification typing kit.Results:1.The percentage of CD4+CD25+CD127lo/-T cells of CD4+T lymphocytes in peripheral blood from patients with cervical cancer and CINⅡ-Ⅲ and contents of cytokine were significantly difference in comparison with that in patients with CINI and in normal controls (P<0.05).2.The CD4+CD25+CD127lo/-T cells percentage in cervical cancer and CIN patients was negatively correlated with IFN-γ (r=-0.585, P=0.000).The CD4+CD25+CD127lo/-T cells percentage in cervical cancer and CIN patients was positive correlated with TGF-β、 IL-10(r=0.690, P=0.000, r=0.827, P=0.000).3. The percentage of CD4+CD25+CD127lo/-T cells of CD4+T lymphocytes in peripheral blood from patients with cervical cancer and contents of cytokine of preoperative were significantly difference in comparison with that in postoperative (P<0.05).4.The expression of Foxp3in CD4+CD25+T cell from patients with cervical cancer were significantly difference in comparison with that in patients with CINI and in normal controls (P<0.05).5. The CD4+CD25HighT cells percentage in cervical cancer and CINⅡ-Ⅲ patients was positive correlated with Foxp3mRNA (r=0.853, P=0.000; r=0.586, P=0.000).6. Treg was positive correlated with HPV (r=0.560, P<0.05) Conclusion:CD4+CD25+CD1271o/-Treg, TGF-β,IL-10and IFN-y participate the process of immune evasion of cervical cancer of Uygurs. HPV was intimate correlated with Treg in cervical cancer of Uygurs.更多还原