阴道微生物与子宫颈HPV感染及子宫颈病变的关系

子宫颈癌是威胁全球女性健康的第四大癌症,年轻女性的子宫颈癌发病率及死亡率呈上升趋势,人乳头瘤病毒(HPV)特别是高危型HPV(HR-HPV)持续感染是导致子宫颈癌前病变及子宫颈癌的主要原因。阴道微生物在女性生殖道健康中发挥重要作用,研究表明阴道微生物失调如乳杆菌数量减少、厌氧菌数量上升、微生物多样性增加等与女性子宫颈HPV感染、子宫颈癌前病变及子宫颈癌之间存在相关性,本文综述了常见阴道微生物与子宫颈HPV感染及子宫颈病变之间的关系并总结其相关机制,为防治子宫颈疾病提供新的切入点。     

HPV联合宫颈细胞学检查在宫颈癌筛查中的应用

持续性高危人乳头瘤病毒（HPV）感染是宫颈癌发生的危险因子。由于宫颈癌筛查技术不断提高,使宫颈癌的发病明显下降。目前许多国家推荐应用HPV DNA检测联合宫颈细胞学检查进行宫颈癌筛查。尽管HPV DNA检测应用于宫颈癌的筛查在临床上显示了明显优势,但由于HPV感染到发展为癌症要经过几十年的潜伏期,即使有HPV感染也不一定发展为宫颈癌。而滥用HPV DNA检测导致的错误诊断将给患者带来不必要的焦虑、额外的开支、过度的治疗。     

基于高危型HPV检测的子宫颈自取样标本固体介质应用的可行性研究

目的:探讨基于高危型人乳头瘤病毒(HPV)检测的子宫颈自取样标本固体介质与传统液体介质的一致性,评价其应用的可行性。方法:选择河北省邢台市平乡县参与子宫颈癌筛查的30～59岁数据资料完整的2027例女性自然人群为研究对象。均经专业培训后行HPV自取样,标本先后应用于固体介质POI卡和传统液体介质,分别采用Cobas 4800 HPV检测(简称Cobas检测)及Seq HPV检测(简称Seq检测)。所有HPV阳性者召回行阴道镜检查及子宫颈活检,以组织病理结果作为诊断金标准,分析自取样固体介质与液体介质采用两种检测方法HPV检测的一致性及检出子宫颈高级别病变的诊断效能。结果:(1)Cobas检测、Seq检测对两种介质HPV阳性率检测结果的一致率分别为96.8%、99.4%,固体介质、液体介质对两种检测方法HPV阳性率检测结果的一致率分别为96.5%、96.9%。(2)任一HPV阳性者255例,组织活检检出子宫颈上皮内瘤变2级及以上(CIN2⁺)29例,子宫颈上皮内瘤变3级及以上(CIN3⁺)共13例。4种检测方法检出CIN2⁺     

HPV联合宫颈细胞学检查在宫颈癌筛查中的应用

持续性高危人乳头瘤病毒(HPV)感染是宫颈癌发生的危险因子.由于宫颈癌筛查技术不断提高,使宫颈癌的发病明显下降.目前许多国家推荐应用HPV DNA检测联合宫颈细胞学检查进行宫颈癌筛查.尽管HPV DNA检测应用于宫颈癌的筛查在临床上显示了明显优势,但由于HPV感染到发展为癌症要经过几十年的潜伏期,即使有HPV感染也不一定发展为宫颈癌.而滥用HPV DNA检测导致的错误诊断将给患者带来不必要的焦虑、额外的开支、过度的治疗.     

宫颈癌机会性筛查人群中高危型HPV感染状况及其与宫颈病变关系的研究

目的:分析宫颈癌机会性筛查人群中高危型HPV感染情况及其与宫颈病变的关系。方法:统计2015年6月至2016年11月于北京大学第三医院行宫颈癌机会性筛查人群中HPV 16型、18型及其他12种HPV亚型的阳性率,计算高危型HPV阳性转诊阴道镜人群中发生CINⅡ及以上(CINⅡ~+)的比率及CINⅡ~+人群中HPV的感染构成情况。结果:宫颈癌机会性筛查人群中,高危型HPV总阳性率为19.0%,HPV 16阳性率为5.0%,HPV 18阳性率为1.6%,其他12种HPV阳性率为15.2%。HPV 16阳性、HPV 18阳性及其他12种HPV阳性患者TCT分流检测≥ASCUS者均转诊阴道镜病理活检,其发生CINⅡ~+的比率依次为55.2%、35.5%及38.9%;发生CINⅢ及以上(CINⅢ~+)的比率依次为25.3%、7.7%及6.9%。CINⅡ~+中45.0%为其他12种HPV感染、46.3%为HPV16感染、8.7%为HPV 18感染,其余病例为HPV阴性。结论:宫颈癌机会性筛查人群中,CINⅡ~+患者HPV 18阳性所占比率较低,其他12种HPV阳性所占比率高。需对各亚型阳性率进行细致分型以协助评估发病风险。转诊阴道镜人群中,HPV 18阳性者及其他12种HPV阳性患者TCT检查为ASCUS/LSIL者发生CINⅡ~+风险相对较低,可考虑通过行分流检测以提高筛查的特异性。     

初善仪和子宫颈液基细胞学及人乳头瘤病毒检测在子宫颈疾病筛查中的应用价值研究

目的:研究子宫颈癌筛查系统初善仪(TS)、子宫颈液基细胞学(TCT)和人乳头瘤病毒(HPV)检测在子宫颈疾病筛查中的应用价值,寻找更为有效的子宫颈癌筛查方案。方法:选取自2018年5月至2019年1月就诊于郑州大学第三附属医院妇科门诊进行子宫颈疾病筛查的已婚女性共315例,分别行TS、HPV和TCT检查,任一结果阳性均行阴道镜下子宫颈组织活检术,以子宫颈组织病理结果作为诊断标准,评价3种筛查方法的灵敏度、特异度、假阴性率、假阳性率和Kappa值。结果:①3种筛查方法单独应用时,与TCT、HPV相比,TS灵敏度、假阴性率差异无统计学意义,但特异度、假阳性率差异有统计学意义(P0.05);TS的Kappa值最高为0.59(P0.05)。②HPV+TS与TCT+TS联合筛查的灵敏度、特异度、假阳性率及假阴性率比较差异均无统计学意义(P0.05);Kappa值分别为0.73、0.57、0.00,TS联合HPV筛查结果与病理结果一致性显著。③HPV阳性患者再行TS检查,筛查特异度较HPV单独筛查时提高至93.33%(P0.05);TCT阳性患者再行TS检查,TS筛查灵敏度、特异度均有提高,但与TCT单独筛查比较,差异无统计学意义(P0.05);两种筛查方案的Kappa值分别为0.79、0.51(P0.05)。结论:3种筛查方法单独应用时TS的特异度及与病理结果一致性均较其他两种方法好。TS联合HPV筛查结果与病理结果一致性显著提高。HPV或TCT检测阳性患者再行TS筛查时均能提高筛查的特异度及病理结果一致性。     

298例宫颈病变中高危型HPV亚型分布情况分析

目的 统计分析宫颈癌及癌前病变中高危型人乳头瘤病毒（HPV）亚型分布情况。方法 选取298例无锡市锡山区9个乡镇筛查出的宫颈癌及癌前病变患者,统计分析本地区宫颈癌及癌前病变中高危型HPV亚型分布情况。结果 298例宫颈癌及宫颈病变中有220例单一型高危型HPV感染,其中前三位亚型感染为16型、58型,52型和33型并列第三,18型排名第六位,无59型感染。CINⅠ级病变中最多感染为四重感染,CINⅡ级病变最多为三重感染,CINⅢ及宫颈癌均最多为双重感染,且各仅有1例。结论 不同级别宫颈病变均以单一型感染为主,且随着病变的程度加重,单一型感染呈上升趋势;多重感染并不促进宫颈病变的发生发展,病变的严重程度与高危型HPV感染的数量无关。     

HPV感染的诊断及其临床意义

人乳头瘤病毒（human papilloma virus，HPV）与宫颈病变的发生有着密切的关系。目前已鉴定的HPV型别约有130余种，不同型别甚至不同亚型的HPV病毒其致病力不同，依据HPV病毒与宫颈病变的关系可将其分为高、中、低危型。其中高危型HPV，如HPV16，18，45，46常引起宫颈上皮内瘤样病变（CIN）和宫颈癌。因此，对HPV感染进行检测和分型在宫颈癌筛查及癌前病变病情随访监测中有着重要的意义。     

HPV单一感染与多重感染对宫颈病变影响的比较

目的：探讨单一高危HPV亚型与多重高危HPV亚型感染对宫颈病变的影响。方法：选取2010年1月—12月在天津医科大学总医院妇科门诊就诊的有性生活史并出现临床症状的患者137例,比较患者细胞学、病理学结果出现的概率;比较单一高危HPV亚型与多重高危HPV亚型感染对宫颈病变影响的差别。结果：①在全部感染病例中,以20～49岁者居多,并且随着年龄的增长,感染率呈现下降趋势。②感染病例中,高危型别以HPV16感染率最高。③在宫颈高级别病变中,以HPV16感染率最高。④单一高危与多重高危HPV感染相比较,宫颈细胞学和病理学结果差异无统计学意义。⑤高低危混合HPV感染中低级别病变概率高于单纯高危HPV感染,而高级别病变则相反。结论：①与单一高危HPV亚型相比,多重高危HPV亚型感染对宫颈病变严重程度的影响不明显。②与单纯高危HPV亚型感染相比,高低危HPV亚型混合感染更多表现为低级别宫颈病变。     

TLRs在宫颈HPV16持续性感染及宫颈病变中的作用研究

目的:探讨Toll样受体(TLRs)在宫颈人乳头瘤病毒(HPV)16持续性感染及宫颈病变中的作用。方法:以PCR法检测宫颈脱落细胞中TLR3、TLR7、TLR8、TLR9及干扰素β(IFN-β)基因在宫颈HPV16持续性感染及宫颈病变中的表达,并分为HPV16持续感染组与HPV16非持续感染组及宫颈炎症及CINⅠ组、高级别CIN和原位癌组、宫颈浸润癌组进行比较分析。结果:1在HPV16持续感染组与HPV16非持续感染组中,TLR3 DNA表达量分别为0.622±0.160、0.696±0.155,干扰素β(IFN-β)DNA表达量分别为0.640±0.169、0.735±0.151,TLR3、IFN-βDNA在HPV16持续感染组表达量低于HPV16非持续感染组(t=6.574,P=0.011;t=4.378,P=0.038);TLR3 DNA与IFN-βDNA呈正相关(r=0.693,P0.001),TLR 9DNA与IFN-βDNA呈负相关(r=-0.384,P0.001)。2在宫颈炎症及CINⅠ组、高级别CIN和原位癌组、宫颈浸润癌组中,TLR3DNA表达量分别为0.661±0.159、0.606±0.143、0.507±0.160,IFN-β表达量分别为0.722±0.169、0.659±0.144、0.483±0.143,TLR7DNA表达量为0.737±0.198、0.754±0.206、0.864±0.167,TLR9DNA表达量为0.531±0.172、0.606±0.192、0.701±0.200。TLR3、IFN-βDNA表达量随宫颈病变进展而逐渐下降(F=7.983,P0.001;F=12.163,P0.001),而TLR7、TLR9 DNA表达量则随宫颈病变进展而逐渐增高(F=3.647,P=0.028;F=9.415,P0.001)。结论:TLRs可能参与宫颈局部免疫系统对HPV16的清除过程,TLR3表达减低可能通过下调IFN-β表达而影响宫颈局部免疫功能,从而导致HPV16持续性感染。TLR3、TLR7、TLR9异常表达在宫颈病变发生及进展中可能发挥重要作用。     

不同女性人群生殖道人乳头瘤病毒感染状况分析

目的:调查不同女性人群生殖道人乳头瘤病毒(HPV)感染状况,为制定不同人群宫颈癌筛查策略提供理论依据。方法:选择2010年1月至2015年12月在重庆市妇幼保健院妇产科门诊体检和就诊的3376例妇女,分为5组:普通人群组770例,妇科疾病患者组1069例,宫颈上皮内瘤变(CIN)患者组560例,不孕症患者组648例,产褥期妇女组329例。采用HPV基因分型技术进行HPV检查,分析HPV DNA亚型感染状况。结果:(1)共检出HPV 22个型别,最常见类型依次为6型(14.81%,500/3376),16型(14.16%,478/3376),52型(12.59%,425/3376),58型(11.91%,402/3376),43型(6.66%,225/3376),未检测出HPV 44型。(2)不同人群HPV总阳性率及高危型阳性率不同,由高到低依次为CIN患者组[89.46%(501/560),85.89%(481/560)],不孕症患者组[66.67%(432/648),58.80%(381/648)],妇科疾病患者组[62.77%(671/1069),55.57%(594/1069)],普通人群组[24.03%(185/770),19.22%(148/770)],产褥期妇女组[13.07%(43/329),10.03%(33/329)]。(3)不同人群中HPV DNA亚型分布有差异,CIN患者组及不孕症患者组以高危型HPV感染为主,感染HPV型别构成比相似,最常见类型均为HPV16、58、52型,且HPV多重感染百分率高于其他组(P0.01),但妇科疾病患者组总体高危型HPV感染率及多重感染率明显高于普通人群及产褥期妇女组。(4)CIN患者组中随CIN病变级别增高,HPV总阳性率及高危型HPV阳性率皆增高,差异有统计学意义(χ~2总=57.49,P0.01;χ~2高危型=58.40,P0.01)。三重及以上HPV感染在高级别病变(CINⅡ、Ⅲ)中更常见(χ~2=8.96,P0.05),在CINⅡ、CINⅢ组中HPV16型检出率高于CINⅠ组,差异有统计学意义(χ~2=33.24,P0.01)。结论:不孕症患者及妇科疾病患者是高危型HPV感染及CIN的高危人群,重视机会性筛查是提高CIN检出率的有效手段。不同女性人群HPV感染状态不同,应针对不同女性人群制定特异筛查模式。     

1955例武汉市农村妇女HPV感染状况与危险因素分析

目的:探讨武汉市农村妇女HPV在宫颈炎、宫颈上皮内瘤变(CIN)、宫颈癌中的感染分布情况及HPV感染危险因素。方法:在2011年武汉市20万农村妇女宫颈癌初筛阳性患者中随机抽取1955例,采用飞行时间质谱检测法对宫颈刷片进行HPV分型测定,分析HPV在宫颈炎、CIN、宫颈癌中的感染情况。结果:1955例宫颈刷片标本中发现HPV感染1115例,感染率为57.03%。HPV在不同宫颈疾病中的感染率不同,HPV在宫颈炎中的感染率为40.72%,CIN中为89.64%,宫颈癌中为92.86%,差异有统计学意义(P<0.01);HPV感染患者中,单纯高危型及单纯低危型HPV感染所占比例分别为92.74%、3.23%,差异有统计学意义(P<0.01)。1955例患者中HPV 16型感染率最高(18.16%),其次是HPV 58型(13.35%)、HPV52型(11.00%)、HPV 33型(5.68%)。1955例患者中单一亚型感染率为41.43%,双重及多重感染率为15.60%。不同初婚年龄、临床症状、宫颈癌家族史及是否熬夜等患者HPV感染率比较,差异有统计学意义(P<0.05)。结论:武汉市农村妇女高危型HPV感染是宫颈癌及CIN发生的必要因素。初婚年龄较早、有宫颈癌家族史、经常熬夜等为HPV感染的危险因素。     

Self-collection for high-risk HPV detection in Brazilian women using the careHPV™ test

Objective

Cervical cancer is the second most common cancer among Brazilian women. High-risk human papillomavirus (hr-HPV) persistence is the primary cause of cervical neoplasia. Early detection of hr-HPV is important for identifying women at risk for developing cervical lesions. Approximately 85% of new cases of cervical cancer worldwide and 50% of the total cervical cancer deaths occurred in developing countries. Here, a new methodology to support a cervical cancer screening program was evaluated in women from various Brazilian regions.

Methods

Two thousand women aged 18–77 years were enrolled in an opportunistic cervical cancer screening program and were randomized into self-vaginal or health professional-guided cervical sampling groups. The Qiagen careHPV™ test was performed on all samples. Pap tests were performed on all women using liquid-based cytology.

Results

Positive hr-HPV results were obtained in 12.3% (245/2000) of women; similar rates were observed in self- or health professional-collected samples. Eighty-nine percent (1719/2000) of cervical cytologies classified as normal were negative to hr-HPV. Among the cytological samples, 36.6% classified as ASC-US + were positive to hr-HPV, 78.8% were LSIL and 75.0% were HSIL.

Conclusions

Self-sampled and health professional-sampled vaginal/cervical specimens did not differ in their rates of detection of hr-HPV. Therefore, HPV DNA testing in self-sampled vaginal cells is an alternative to primary screening in low-resource settings.     

宫颈癌患者外周血HPV16抗原特异性CD8~+细胞毒性T细胞的检测及意义

目的:初步探讨人乳头瘤病毒16型(HPV16)阳性宫颈癌患者细胞免疫状况,了解其外周血抗原特异性CD8~+细胞毒性T细胞的水平.方法:采取免疫荧光标染重组MHC Ⅰ类分子-肽五聚体技术,运用流式细胞术定量检测患者和正常人外周血CD8~+细胞毒性T细胞、记忆性细胞毒性T细胞、活化细胞毒性T细胞及HPV16抗原特异性CD8~+细胞毒性T细胞.结果:宫颈癌患者CD8~+细胞毒性T细胞计数、记忆性细胞毒性T细胞计数、体内抗原特异性CD8~+细胞毒性T细胞计数和对照组比较差异无统计学意义,但活化细胞毒性T细胞计数低于对照组(P=0.322),体外经抗原肽刺激后抗原特异性CD8~+细胞毒性T细胞计数高于对照组(P=0.0068).结论:HPV16 阳性宫颈癌患者外周血中活化细胞毒性T细胞数目减少,表明患者细胞免疫功能受到一定的影响,通过抗原表位有效的诱导是提高患者的特异性细胞免疫功能的重要途径,运用五聚体技术检测宫颈癌患者抗原特异性细胞毒性T细胞,可初步反映患者特异性细胞免疫状态并有利于进行复发和预后分析.     

Role of HPV DNA testing in modern gynaecological practice

The identification of some types of human papillomavirus (HPV) as necessary, but not sufficient, cause of cervical cancer has suggested the use of HPV testing in cervical cancer prevention. A large number of studies has provided evidence supporting its application (1) as primary screening test, (2) for triaging borderline cytology, (3) for follow-up after positive primary test but no abnormal histology and (4) as a test of cure. They also allowed a reasonably good definition of the appropriate policies and protocols, leading to the delivery of evidence-based guidelines resulting from a systematic review of the literature. In this chapter, we present a critical analysis of the recommendations of the main European and North American guidelines relative to industrialised countries.     

宫颈鳞癌中Cyclin E的表达和HPV16感染的相关研究

目的探讨Cyclin E在宫颈鳞状细胞癌中的表达及其与HPV16感染的关系.方法应用免疫组化S-P法对34例宫颈鳞状细胞癌组织进行Cyclin E与HPV16的检测,并与宫颈不典型增生和正常宫颈组织进行比较.结果正常宫颈组织无Cyclin E表达,宫颈不典型增生组织仅有少量袁达(11.11%),宫颈鳞状细胞癌有58.52%表达(P＜0.001),且与HPV16感染有明显相关性(P＜0.05).结论高危HPV感染是宫颈癌的重要病因,HPV感染通过多种途径改变了细胞的周期性调控,诱发细胞周期GI-S期检测点异常,引起细胞恶性增生,HPV感染与Cyclin E异常表达在宫颈癌的发生发展中起重要作用.     

Methylation of HPV16 genome CpG sites is associated with cervix precancer and cancer

Objective.

Invasive cervix cancer (ICC) is the second most common malignant tumor in women. Human papillomavirus 16 (HPV16) causes more than 50% of all ICC and is a major cause of cervix intraepithelial neoplasia (CIN). DNA methylation is a covalent modification predominantly occurring at CpG dinucleotides. Such epigenetic modifications are associated with changes in DNA-protein interactions and gene activation. This study examined the association of viral and host genomic methylation patterns and cervix neoplasia.

Methods.

Exfoliated cervical lavage samples positive for HPV16 from women with and without cytomorphic changes of infection (n = 46), CIN2 (n = 12), and CIN3+ (n = 27) were used to interrogate the methylation patterns of the HPV16 L1 gene and upstream regulatory region (URR), five host nuclear genes (TERT, RARB, DAPK1, MAL, and CADM1), and mitochondrial DNA (mtDNA). DNA isolated from exfoliated cervicovaginal cells was treated with bisulfite, specific regions of the viral and host genome were PCR amplified and CpG methylation was quantified using EpiTYPER and pyrosequencing.

Results.

Methylation at 14 of the tested CpG sites within the HPV16 L1 region were significantly higher in CIN3+ compared to HPV16 genomes from women without CIN3+. In contrast, 2/16 CpG sites in HPV16 URR, 5/5 in TERT, 1/4 in DAPK1 and 1/3 mtDNA, and 2/5 in RARB were associated with increased methylation in CIN3+.

Conclusions.

These results indicate that increased methylation of CpG sites in the HPV16 L1 ORF is associated with CIN3+ and, thus, may constitute a potential biomarker for precancerous and cancerous cervix disease.     

Persistent HPV infection in postmenopausal age women.

OBJECTIVE: Persistence of human papillomavirus (HPV) is associated with an increased risk of developing cervical SIL and cancer in young women. Because this association in older, postmenopausal age women has received little attention, we evaluated persistence of HPV among women in this age group. METHODS: Women (n=105) ages 45-64 were examined annually for 7 years to evaluate HPV in cervical cytologic specimens. PCR, dot blot hybridization and DNA sequencing were used to detect HPV types. RESULTS: The cumulative prevalence of HPV was 34%, and 24% had HPV high-risk oncogenic types which are associated with genital cancers. The most common oncogenic types were HPV-16 (72%) and HPV-31 (16%). The persistence rate of HPV infection was 16%. No specific risk factors were associated with repeat viral positivity. CONCLUSION: Postmenopausal women are infected with persistent oncogenic HPV at a substantial rate, supporting the need for continued screening in postmenopausal women to detect preneoplastic genital lesions.