宫颈癌组织端粒酶表达与人乳头瘤病毒感染的相关性

目的 探讨宫颈癌组织中端粒酶活性表达与人乳头瘤病毒（human papillomavirus，HPV）感染的关系。方法 采用端粒重复序列扩增（telomere repeat amplification protocal，TRAP）-银染法对45例宫颈癌、30例慢性宫颈炎及20例正常宫颈组织的端粒酶活性进行检测。同时，采用多聚酶链反应（PCR）技术测定上述组织HPV、HPV-16型及HPV-18型的感染情况。结果 宫颈癌组织端粒酶阳性率、HPV和HPV-16检出率高于慢性宫颈炎和正常宫颈组织（P〈0．05）；宫颈癌组端粒酶表达与HPV感染（φ=0．44，P〈0．01）、HPV-16型感染（φ=0．36，P〈0．05）有关联性，与HPV-18型感染未发现关联性（P〉0．OS）。结论 端粒酶的表达、HPV感染包括HPV-16型感染与宫颈癌发生密切相关。     

用聚合酶链反应检测喉鳞癌细胞内人乳头状瘤病毒DNA

用聚合酶链反应（ＰＣＲ）技术检测喉鳞状细胞癌患者的经福尔马林固定、石蜡包埋的组织标本２８例。从石蜡包埋的癌组织提取的人乳头状瘤病毒ＤＮＡ（ＨＰＶＤＮＡｓ）用ＨＰＶ－１６，１８试剂盒进行ＰＣＲ扩增。其中５例（１８％）扩增阳性。声带癌ＨＰＶ－１６，１８的阳性率较其它部位的高（占阳性８０％）。结果提示：ＨＰＶ－１６，１８的感染在喉癌的发生中起到一定的作用。声门区的喉鳞癌与宫颈癌的发生类似，与ＨＰＶ－１６，１８的感染有密切的联系。     

高危型HPV感染与食管鳞癌组织中HLA-G表达的相关性研究

目的研究人乳头瘤状病毒(HPV)在新乡地区食管鳞癌、癌旁组织中的感染情况和人类白细胞抗原系统G(HLA-G)表达的关系。方法采用基因芯片技术检测114例食管鳞癌组织及癌旁食管正常组织中HPV亚型感染情况,免疫组化SP法检测HLA-G的表达。结果 114例食管鳞癌组织及癌旁食管正常组织中HPV感染阳性率分别为63.2%和6.0%,HLA-G的检出率分别为51.8%、0%,差异均有统计学意义(P<0.05);食管鳞癌存在HPV16、18和52三种亚型感染,其中HPV16感染率最高,HPV52感染率最低。结论新乡地区食管鳞癌组织以HPV16、18和52亚型感染为主;HPV感染和HLA-G蛋白表达与食管鳞癌分化程度正相关;HPV感染食管鳞癌组织与HLA-G蛋白表达存在一定的相关性,可能提示与食管鳞癌发生有较密切关系。     

人乳头状瘤病毒和EB病毒在子宫颈上皮内瘤变中的表达

目的:探讨人乳头状瘤病毒(HPV)和EB病毒(EBV)在子宫颈上皮内瘤变(CIN)中的表达.方法:采用原位杂交对130例CIN(CIN1 50例,CIN2 40例,CIN3 40例)、子宫颈癌10例、子宫颈湿疣样病变10例、子宫颈正常黏膜10例的HPV(高危型16/18、低危型6/11)和EBV进行检测.结果:HPV16/18在CIN1、CIN2、CIN3的阳性表达率分别是44.0%、37.5%和62.5%,子宫颈癌为8/10,子宫颈湿疣样病变为10/10,子宫颈正常黏膜为3/10,差异有统计学意义(P<0.01).HPV6/11在CIN1、CIN 2、CIN3的表达率分别是36.0%、52.5%和70.0%,在子宫颈癌中的表达是6/10,子宫颈湿疣样病变9/10,子宫颈正常黏膜3/10,差异有统计学意义(P<0.01).HPV16/18和HPV6/11在CIN1、CIN2、CIN3中联合表达率分别为26.0%、22.5%和50.0%,子宫颈癌为6/10,子宫颈湿疣样病变为7/10,子宫颈正常黏膜为2/10,差异有统计学意义(P<0.01).EBV在CIN1、CIN2、CIN3的阳性表达率分别是24.0%、12.5%和22.5%,子宫颈癌为3/10,子宫颈湿疣样病变为0/10,子宫颈正常黏膜为0/10,差异无统计学意义(P>0.05).结论:HPV感染是引起CIN的主要致病因子,临床上往往是多种亚型混合感染.EB病毒在CIN和子宫颈癌表达的意义仍有争议.     

端粒酶活性在人乳癌中的表达

目的：比较端粒酶活性在乳腺良,恶性病变中的异同,探讨其在乳癌诊断中的意义和与普遍认定的预后指标间的关系。方法：用端凿重复扩增（ＴＲＡＰ）检测了５４例乳腺癌、４４例乳腺良性病变组织的端粒酶活性。结果：５４例乳癌中４９例（９０．７４％）显示端粒酶活性,１例纤维腺瘤有弱端粒酶活性。端粒酶活性与肿瘤大小及分期有关,而与腋淋巴结转移及雌,孕激素受体之间无关。结论：端粒酶活性见于绝大多数浸润性乳癌及极少数纤维     

人乳腺浸润性导管癌端粒酶活性的检测

目的比较乳腺浸润性导管癌及乳腺良性病变组织端粒酶活性的异同,探讨端粒酶活性在恶性肿瘤诊断中的意义。方法用端粒酶重复扩增法（ＴＲＡＰ）检测了６１例乳腺浸润性导管癌及１４例乳腺良性病变组织端粒酶活性。结果６１例中４９例（８０％）显示端粒酶活性,端粒酶活性与浸润性导管癌的分级、肿瘤大小、淋巴结转移及肿瘤组织雌激素受体及孕激素受体表达无关。５例乳腺囊腺病无一例端粒酶活性,９例乳腺腺瘤中４例端粒酶弱阳性。结论端粒酶活性见于绝大多数乳腺浸润性导管癌及极少数的乳腺腺病中,该酶活性可能在乳腺癌的发生发展中起重要作用     

西安交通大学第一附属医院8581名妇女生殖道人乳头瘤病毒感染筛查

目的 调查陕西地区妇女生殖道人乳头瘤病毒(HPV)感染情况,了解本地区妇女生殖道HPV感染特征。方法 采用PCR+导流杂交技术检测2009年2月至2012年12月在西安交通大学第一附属医院妇科就诊的8581名陕西地区妇女生殖道HPV-DNA,必要时进行宫颈细胞学或组织学检查。结果 8581名陕西地区妇女中,生殖道HPV感染率为33.06%,高危型HPV(HR-HPV)感染率为30.08％,复合型别感染者占27.18%;最常见感染型别依次为HPV-16、HPV-52、HPV-58、HPV-6和HPV-18。HPV感染存在年龄差异,感染高峰年龄为25岁以下和51岁以上。HR-HPV感染者中宫颈组织学正常、轻度上皮内瘤变(CINⅠ)、中度上皮内瘤变(CINⅡ)、重度上皮内瘤变(CINⅢ)和宫颈癌(CC)组中,感染率分别为88.72%、97.48%、97.35%、95.88%和99.23%;常见致癌型别依次为HPV-16、HPV-18、HPV-58、HPV-52和HPV-33。HPV-16、18和58共同引起94.62%的宫颈癌。结论 陕西地区妇女生殖道HPV感染比较普遍,最常见的感染型别是HPV-16、HPV-52、HPV-58、HPV-6和HPV-18,常见致癌型别依次为HPV-16、HPV-18、HPV-58、HPV-52和HPV-33。HPV-16、18和58引起94.62%的宫颈癌。高危型HPV感染和宫颈病变显著相关,HR-HPV复合感染不是宫颈病变的危险因素。     

宫颈尖锐湿疣组织中HPV分型与端粒酶活性的研究

目的由于宫颈尖锐湿疣(CA)皮损中HPV高危型感染常与其癌变有关，而端粒酶活性增高多见于恶性疾病。该研究探讨宫颈CA皮损中HPV分型与端粒酶活性的关系。方法采用荧光定量PCR检测HPV病毒分型，端粒重复序列扩增文件-酶标法(TRAP—ELISA)检测端粒酶活性。结果尖锐湿疣患者皮损中端粒酶活性明显高于正常皮肤组织；24例(40．0％)HPV16／18型阳性，34例(56．7％)HPV6／11型阳性，2例未检测到HPV；HPV16／18型感染的皮损中端粒酶活性明显高于HPV6／11型感染者。结论认为端粒酶活性与HPV型别有关，可能与HPV病毒协同参与CA发病，HPV16／18型阳性者在临床可能有癌变倾向，抑制端粒酶活性可能可以阻碍疣体生长，为临床治疗提供一个新的方向。     

酶的活性及其与凹空细胞分型的关系

目的 研究宫颈上皮内瘤样病变（CIN）及宫颈癌的组织和脱落细胞中端粒酶的活性变化,探索端粒酶激活与凹空细胞分型及人乳头状瘤病毒（HPV）感染之间的关系。方法 采用端粒酶重复扩增技术（TRAP）,检测24例正常宫颈、34例宫颈炎症、78例CIN及16例宫颈癌的宫颈组织和脱落细胞中端粒酶的活性。同时采用双盲法检测上述宫颈病变组织中凹空细胞的分布,并将其分为Ⅰ型和Ⅱ型。结果 组织中端粒酶阳性率分别为：正常宫颈组织为16.7%（4／24）,炎症组织为17.6%（6／34）,CIN1组织为51.6%（16／31）,CIN2组织为72.0%（18／25）,CIN3组织为86.4%（19／22）,宫颈癌组织为100％（16／16）。端粒酶阳性率CIN明显高于正常宫颈及炎症组织（P＜0.01）;宫颈癌组织明显高于CIN组织（P＜0.05）;CIN3高于CIN1（P＜0.05）;而正常宫颈与炎症组织比,CIN1与CIN2比,CIN2与CIN3比较,差异均无显著意义（P＞0.05）。相应的宫颈脱落细胞中端粒酶阳性率同组织标本比,差异无显著意义（P＞0.05）。含Ⅰ型、Ⅱ型凹空细胞的宫颈病变组织中端粒酶阳性率分别为66.7%和100％,两者差异有显著意义（P＜0.01）。结论 （1）端粒酶的激活与宫颈癌及CIN的发展有密切关系。端粒酶可作为宫颈癌及CIN的标志物。（2）宫颈脱落细胞可以替代组织标本进行端粒酶活性检测。（3）端粒酶的激活与Ⅱ型凹空细胞的出现及HPV16／18感染有密切关系。     

胃良恶性病变组织中端粒酶的活性研究

目的:探讨胃癌和胃良性病变中端粒酶的活性及其临床意义.方法:应用TRAP-PCR-ELISA方法检测36例胃癌组织,20例胃溃疡组织,10例胃炎组织和20例正常人胃黏膜组织中端粒酶的活性.结果:胃癌组织中端粒酶阳性率分别为80.6%(29/36),胃良性病变组织中端粒酶阳性率为10%(其中2例胃溃疡和1例胃炎组织标本检测到端粒酶活性),正常人胃黏膜组织未检测到端粒酶阳性.胃癌组织与胃良性病变组织中端粒酶阳性率比较,差异有统计学意义(P＜0.01).结论:端粒酶活性是判断胃良恶性肿瘤的一个重要指标,对胃良性病变患者端粒酶检测阳性者应加强随访.     

Correlation between physical status of human papilloma virus and cervical carcinogenesis

The prevalence of human papilloma virus (HPV)-16 in patients with cervical cancer,the physical status of HPV-16 in patients with cervical lesions,and the role of HPV-16 integration in cervi-cal carcinogenesis were investigated.HPV genotyping was performed by using PCR approach with the primer GP5+/GP6+ and type-specific primer on biopsy specimens taken operatively from 198 women.Multiple PCR was done to detect physical status of HPV-16 in a series of cervical liquid-based cytology samples and biopsy specimens obtained from different cervical lesions with HPV-16 infection,includ-ing 112 specimens with cervical cancer,151 specimens with CINⅠ,246 specimens with CINⅡ and 120 specimens with CINⅢ.The results showed that there were 112 cervical cancer samples (56.57% of total cervical cancer patients) with HPV-16 infection.The frequency of HPV-16 pure integration was 65.18% (73/112),56.57% (47/120),23.58% (58/246) and 7.95% (12/151) in cervical cancer,CINⅢ,CINⅡand CINⅠ patients respectively.In situ hybridization was performed on some paraffin-embedded sections of CINⅡ,CINⅢ and cervical cancer to verify the physical status of HPV-16 infection.Sig-nificant difference was observed between cervical cancer and CINⅠ,CINⅡ,CINⅢ in the frequency of HPV-16 integration (P<0.01).It is suggested that HPV-16 is the most prevalent type and is associated with cervical cancer.In the case of HPV-16 infection there are close associations between the severity of cervical lesions and the frequency of HPV-16 integration.The application of testing HPV genotyping and physical status based on detection of HC-Ⅱ HPV DNA would be in favor of predicting the progno-sis of cervical precancerosis and enhancing the screening accuracy of cervical cancer.     

Telomerase activity in cervical intraepithelial neoplasia

Background It was reported that telomerase expression is closely associated with cellular immortality and cancer. This study was designed to investigate the relationship between telomerase expression and the carcinogenesis of cervical cancer, the possible use of telomerase as a marker of cervical intraepithelial neoplasia (CIN) progression or regression, and the natural history of CIN.Methods Telomeric repeat amplification protocol (TRAP) assay was used to measure telomerase activity in cervical scrapings and biopsy samples obtained from 105 cases affected with various cervical conditions, including chronic cervicitis (n =20), CIN (n =64, 16 cases of CIN Ⅰ , 20 cases of CIN Ⅱ, and 28 cases of CIN Ⅲ）, and invasive squamous cell carcinoma ( n =21 ).Results In exfoliated cell samples, telomerase activity was detected in 5 of 20 (25.0%) cases of cervicitis, 10 of 16 (62.5%) cases of CIN Ⅰ , 11 of 20 (55.0%) cases of CIN Ⅱ, 23 of 28 (82. 1%) cases of CIN Ⅲ, and 13 of 21 (61.9%) cases of carcinoma. In cervical biopsy samples, telomerase activity was detected in 6 of 20 (30.0%) cases of cervicitis, 8 of 16 (50.0%) cases of CIN Ⅰ, 9 of 20 (45. 0%) cases of CIN Ⅱ, 27 of 28 (96.4%) cases of CIN Ⅲ, and 20 of 21 (95.2%) cases of carcinoma. Telomerase activation was significantly higher in CIN samples than in cervicitis samples.Telomerase activity was detected at similar frequency in samples from cervical scrapings and cervical biopsies.Conclusion These results seem to suggest that telomerase expression may be associated with carcinogenesis of the cervix. TRAP assay of cervical scraping samples could be used to monitor and predict the development of CIN in clinical practice.     

高危型人乳头状瘤病毒在子宫颈癌组织和外周血表达的意义

目的: 分别检测子宫颈癌组织和外周血标本中人乳头状瘤病毒(HPV)(16型,18型),探讨HPV感染与子宫颈癌的关联性以及在肿瘤微转移中的表达。方法: 采用荧光定量PCR(FQ-PCR)和逆转录PCR (RT-PCR)技术检测24份子宫颈癌组织及其治疗前后48份外周血标本,慢性子宫颈炎组织及其外周血标本各20份。结果: 24份子宫颈癌组织(Ⅰ~Ⅱb期)中HPVDNA 16型阳性率为66.7%,HPV DNA 18型阳性率为29.2%,两型均阳性的阳性率为8.3%;48份外周血标本HPV mRNA 16型阳性率为2.1%;20份慢性子宫颈炎组织HPV DNA 16型阳性率为15.0%,HPV18型为阴性,其外周血均为阴性。子宫颈癌HPV DNA 16型组与慢性子宫颈炎HPV DNA 16型组比较差异有统计学意义(P<0.005~P<0.05)。结论: 高危型HPV感染与子宫颈癌发生高度相关。外周血HPV检出率低,临床价值有待进一步研究。     

DNA sequences of human papillomavirus types 11, 16, and 18 in lesions of the uterine cervix in the west of Scotland

Punch biopsy specimens of the cervix were examined both histologically and for the presence of human papillomavirus (HPV) DNA sequences. The presence of HPV DNA sequences was sought with the Southern blot technique using radioactively labelled HPV-6, 11, 16, and 18 DNA probes, both together and separately. Twenty six biopsy specimens were examined. Histological examination showed cervical intraepithelial neoplasia grade 2 or 3 in 16 specimens, viral changes (koilocytosis) in four, and inflammation or a normal appearance in three. Eleven specimens were negative for HPV DNA sequences, 10 contained HPV-16 DNA, four contained HPV-18 DNA, and one contained both HPV-18 and HPV-11 DNA. Episomal HPV-16 DNA was detected in one case of cervical intraepithelial neoplasia grade 3 and in five cases of cervical intraepithelial neoplasia grade 2/3 with koilocytosis; and episomal HPV-18 DNA was found in two specimens classed as cervical intraepithelial neoplasia grade 2/3, one of which also contained HPV-11 DNA, and in one specimen that showed viral changes alone. Integrated HPV DNA was found in six specimens (four with HPV-16 DNA and two with HPV-18 DNA), including two cases of chronically inflamed cervix with no histological evidence of viral infection or cervical intraepithelial neoplasia. Detection of viral DNA in early lesions may identify patients at risk of malignant progression. This is the first report of HPV-18 DNA in cervical intraepithelial neoplasia in Scotland.     

口腔鳞状细胞癌中端粒酶活性的表达

目的 了解端粒酶活性在口腔鳞状细胞癌中的表达情况,并探讨端粒酶活性在口腔鳞状细胞癌演变过程中的作用。方法 对３７例口腔鳞状细胞癌、４０例口腔良性肿瘤及１２例正常口腔组织,采用聚合酶链反应－酶联免疫吸附法检测其端粒酶活性。结果 口腔鳞状细胞癌组的端粒酶活性阳性率明显高于良性肿瘤组和正常组织组。结论 端粒酶可稳定染色体末端的端粒结构,端粒酶活化在口腔肿瘤恶变过程中起了重要作用;端粒酶可作为判断癌变能力     

Comparison of polymerase chain reaction and catalyzed signal amplification in situ hybridization methods for human papillomavirus detection in paraffin-embedded cervical preneoplastic and neoplastic lesions

BACKGROUND: Two molecular methods for HPV genotyping in formalin-fixed, paraffin-embedded tissue were evaluated: in house polymerase chain reaction assay (PCR) with consensus and type-specific primers and a novel procedure of in situ hybridization-a catalyzed signal amplification system (CSA-ISH, Genpoint, DAKO, Glostrup, Denmark). The number of HPV positive cases and detected viral types were compared in cervical biopsies and cone specimens according to histopathological diagnosis. Primer efficiency in detecting various types of HPV by PCR method was evaluated. METHODS: DNA samples (101) were used as a template to amplify with three pairs of consensus (MY09/11, GP5+/6 +, CPI/IIG) and four type-specific HPV primers (HPV-6/11, 18, 16 and 33). The according histological tissue sections were analyzed with CSA-ISH method, using commercial HPV biotinylated probes HPV-6/11, 16/18 and 31/33/51. RESULTS: The degree of concordance for PCR and CSA-ISH was 64.4%. In 63 of 101 samples (62.4%), HPV was detected by PCR, while only 35 (34.7%) were positive using CSA-ISH. CSA-ISH found lower percentages for all HPV types, except HPV-6/11. A lower percentage of positive results in all high-grade lesions was detected by CSA-ISH. Multiple infections were detected by PCR in only one sample and in three samples by CSA-ISH. Detection with My09/11 primers followed by Gp5+/6+ primers, in nested reaction, gave the highest number of positive results: 58 of 63 (92%). None of the samples diagnosed as condylomata planum or CIN I was positive for HPV-6/11 (low risk type), which was detected exclusively in condylomata acuminatum group. CONCLUSIONS: A significantly higher number of positive samples was detected with PCR than with CSA-ISH method. CSA-ISH method should be improved, especially in detecting HPV in high-grade lesions. CSA-ISH may be more accurate in detection of multiple infections. GP5+/6+ in nested reaction after MY09/11 detected the highest number of positive results. Samples diagnosed as benign lesions positive on HPV-X must be monitored as possible candidates for progression. CIN I lesions, which were HPV negative, probably will not progress. This finding may be important in planning therapy and avoiding unnecessary treatment.     

宫颈癌患者血液中人乳头瘤病毒检出率的临床意义

目的:探讨宫颈癌患者血液中人乳头瘤病毒(HPV)DNA检出率对宫颈癌的诊断及病情进展评价的临床意义。方法:应用巢式PCR检测48例宫颈癌,55例宫颈高度病变(HG),6例宫颈低度病变(LG),29例宫颈炎症患者血浆中HPV-16,18DNA。分析各组患者血中HPVDNA检出率的差异,以及检出率与宫颈癌临床分期的相关性。结果:血浆中HPVDNA在宫颈癌组、HG组、LG组及宫颈炎症组中的检出率分别为56.3%、41.8%、16.7%和37.9%,其中HPV-16分别为20.8%、5.5%、0.0%和6.9%;HPV-18分别为50.0%、38.2%、16.7%和31.0%。HPV-16和HPV-18均为阳性者8例,其中宫颈癌Ⅰ期3例,Ⅱ期4例,CINⅢ1例。在9例有淋巴结转移的患者中,6例血浆HPVDNA阳性。血浆HPVDNA在宫颈癌组中检出率高于其他3组,但无统计学差异。结论:宫颈癌患者血中HPV检出率高于CIN和宫颈炎症患者,并且在发生转移的宫颈癌患者中检出率更高,但血浆HPVDNA检出率与宫颈癌临床分期无关。宫颈癌患者血中HPV双重感染率较高。     

450例宫颈活检组织中人乳头瘤病毒的分布特征

[目的]了解阴道镜检查患者的宫颈活检组织中HPV的感染率和亚型分布。[方法]对宫颈脱落细胞学异常的患者450例进行阴道镜下定位活检。活检组织同时行HPV基因分型和病理诊断,HPV基因分型采用斑点杂交方法进行。[结果]在450例宫颈活检患者中,其HPV感染率为65.3%。在HPV-DNA阳性患者中,检出率最高的HPV-16占36.7%,依次HPV-58为22.1%、HPV-18为18.7%、HPV-52为16.0%。在宫颈炎、CINⅠ、CINⅡ、CINⅢ中HPV-DNA阳性率分别为55.6%、69%、82.7%和88.9%。在宫颈炎、CINⅠ、CINⅡ,CINⅢ组织中均检测到HPV-16、-58、-18和-52。[结论]本研究中的宫颈活检组织中不仅检测到HPV-16和-18,还检测到HPV-58和-52感染,后者的发现为开发宫颈癌多价复合型疫苗提供了事实依据。     

新疆维吾尔族宫颈癌组织中HPV—DNA表达及存在状态的研究

目的 探讨高危型人乳头瘤病毒（HR—HPV）在新疆维吾尔族宫颈癌患者中的表达、存在状态及其与宫颈癌的相关性。方法 选择第2代杂交捕获（Hybrid capture,HC2）检测HR—HPV阳性的30例维吾尔族宫颈鳞癌（SCC）患者,利用巢式PCR检测其组织标本中的HPV16-DNA的表达,多重Taqman探针荧光定量PCR检测HPV16～DNA在组织中的存在状态。结果 巢式PCR检测结果 显示宫颈鳞癌患者HPV16-DNA检出率为96．7％,与HC2法的HPV—DNA检出率差异无统计学意义（P〉0．05）;HPV16-DNA在宫颈癌组织中以整合态存在,其整合率为89．7％;HPV16-DNA的表达水平与宫颈癌患者的年龄、有无淋巴结转移无关（P〉0．05）,但其在病理分级高的患者中表达高于病理分级低的患者（P〈0．05）,在临床分期高的患者中表达高于低分期患者（P〈0．05）。结论 HR—HPV感染是导致宫颈癌的主要因素,而HPV16是新疆维吾尔族宫颈癌主要的病毒类型,其在组织中多以整合态存在,在宫颈癌发病中起主要作用,HPV—DNA整合状态与宫颈细胞的恶性转化密切相关。