采用原位杂交法检测生殖系统尖锐湿疣和鳞状细胞癌内乳头瘤病毒的感染

目的 探讨HPV感染与生殖系统尖锐湿疣和鳞状细胞癌的关系。方法 采用HPV6／11、16／18原位杂交试剂盒，对55例生殖系统不同病变中乳头瘤病毒感染状况进行分析检测。结果 20例生殖系统尖锐湿疣19例阳性，其中17例为HPV6／11型，2例为HPVl6／18型。20例生殖系统鳞状细胞癌中，15例阳性，均为HPVl6／18型。10例正常生殖系统鳞状上皮组织和5例外阴白斑组织均呈阴性。结论 HPV6／11多与生殖系统良性疣状病变有关，而HPVl6／18感染多见于生殖系统恶性病变。     

应用组织芯片研究HPV6/11、HPV16/18与子宫颈病变的关系

目的 通过检测低危型HPV6/11及高危型HPV16/18在慢性子宫颈炎、子宫颈尖锐湿疣不伴非典型增生、子宫颈鳞状上皮CIN Ⅰ级、子宫颈鳞状上皮CIN Ⅲ级、子宫颈浸润性鳞状细胞癌五种子宫颈病变中的表达情况,探讨HPV与子宫颈病变的相关性、作用机制及其临床意义.方法 应用组织芯片技术,将150例子宫颈病变患者的标本制成组织芯片,用原位杂交法对其进行6/11型及16/18型HPV的测定,应用SPSS 10.0统计软件进行数据分析.结果 低危型HPV6/11在慢性子宫颈炎、子宫颈尖锐湿疣、子宫颈鳞状上皮CIN Ⅰ级、子宫颈鳞状上皮CIN Ⅲ级、子宫颈浸润性鳞状细胞癌中的阳性表达率分别为:13.33%、90%、33.33%、0、0,其中尖锐湿疣组HPV6/11的阳性表达率显著高于其他各组(均P＜0.001);CIN Ⅰ组与CIN Ⅲ及癌比较差异有统计学意义(P=0.001,＜0.05);运用Spearman相关分析表明,HPV6/11在上述五种病变中的阳性表达率与恶性度呈负相关(rs=-0.370,P＜0.001).高危型HPV16/18在慢性子宫颈炎、尖锐湿疣、CIN Ⅰ级、CIN Ⅲ级、鳞状细胞癌中的阳性表达率分别为0、6.67%、10%、56.67%、76.67%,CIN Ⅲ及鳞癌与其他各组间比较均有统计学意义(P＜0.001),但CIN Ⅲ与鳞癌之间无统计学意义(P=0.10);运用Spearman相关分析表明,HPV16/18在上述五种病变中的阳性表达率与恶性度呈正相关(rs=0.628,P＜0.001).结论 低危型HPV6/11主要引起子宫颈尖锐湿疣及CIN Ⅰ;高危型HPV16/18与CIN Ⅲ及子宫颈浸润性鳞癌关系密切,是引起子宫颈CIN Ⅲ及浸润性癌的主要因素.对HPV在子宫颈病变中检测及分型有助于对子宫颈病变的诊断和监测,尤其是对子宫颈癌的预防、早期诊断及早期治疗具有重要意义.     

HPV感染的形态特征及HPV型别与宫颈癌的临床病理联系

目的观察人乳头瘤病毒（HPV）感染的形态学改变,分析挖空细胞形态、HPV型别与宫颈鳞状细胞癌临床病理参数间的联系。方法对594例宫颈鳞状细胞癌进行了形态学观察,按照WHO标准进行组织学分型及分级。运用SPFIO—PCR技术进行HPVDNA扩增,并采用DEIA（DNA enzyme immunoassay）及LiPA（line probe assay）方法进行DNA检测及分型。结果经检测HPVDNA阳性者581例,其中394例具有HPV感染的组织学特点,即出现典型（104例）或不典型（290例）的挖空细胞,检出率为67．8％。36．2％的角化型及72．2％的高分化宫颈鳞状细胞癌中可见典型挖空细胞,而34．0％的非角化型及50．7％的低分化者中无挖空细胞。在HPV型别检测中,以HPV16阳性最多见为453例,其中高、中、低分化者分别为16例、389例及48例;其次为HPV18共43例,其中中分化者32例,低分化者11例;其他高危型HPV按照阳性数多少依次为HPV31、52、59、58、39、45、33、56、66、68、73,分别为17例、12例、12例、11例、6例、5例、5例、3例、2例、1例、1例;低危型HPV包括6及53各1例,且均同时伴有高危型HPV感染。结果显示,HPV16、HPV18阳性者较其他高危型HPV阳性者发病年龄轻;HPV18阳性者较HPV16阳性者分化程度低;HPV18阳性者较HPV16阳性者无挖空细胞病例所占比例较高（分别为48．8％及31．8％）;患者年龄越大FIGO分期越晚。结论挖空细胞形态与宫颈鳞状细胞癌的分化程度密切相关,典型挖空细胞多出现在分化较成熟的癌组织中,而分化较差者常无挖空细胞。HPV16、18是宫颈鳞状细胞癌最重要且最危险的HPV型别。HPV18阳性可能提示宫颈鳞状细胞癌恶性程度高。     

肺鳞状细胞癌中人类乳头瘤状病毒16/18感染和Bcl-2、Bax蛋白表达的关系

背景与目的病毒和肿瘤的关系越来越引起人们的重视,且我国可能是HPV的高感染区,肺癌中是否存在HPV感染以及其是否与肺癌的发生发展有关仍然未明。本研究旨在探讨肺鳞状细胞癌中HPV16/18型感染状态和Bcl-2、Bax蛋白表达的关系。方法采用原位杂交检测肺鳞状细胞癌中HPV16/18DNA感染状态,应用免疫组化检测Bcl-2、Bax蛋白表达。结果44例肺鳞状细胞癌中,HPV16/18型23例(52.27%)呈整合状态,9例(20.45%)呈大部分游离而少许为整合状态,12例(27.27%)阴性。15例非肿瘤肺组织标本中未检测到HPV16/18型,差异有统计学意义(P<0.001);Bcl-2蛋白20例(45.5%)阳性,其表达与HPV16/18在癌细胞中的存在状态有关(P<0.05)。结论HPV感染可能是部分肺鳞状细胞癌的致癌因素之一,在这部分癌中Bcl-2的表达显著高于HPV阴性和存在游离状态HPV的病例。     

合肥地区食管鳞状细胞癌患者HPV感染调查及HR-HPV感染与预后的关系

目的:对合肥地区食管鳞状细胞癌患者人乳头瘤病毒(HPV)感染情况进行调查分析,并探讨高危型HPV(HR-HPV)感染与患者预后的关系。方法:选取2013年06月至2019年02月本院收治的384例合肥地区食管鳞状细胞癌(包括食管鳞状上皮高级别上皮内瘤变伴癌变)患者,均行HPV检测。统计分析纳入研究患者HPV感染情况,另手术/放化疗治疗后随访1年,根据预后情况将患者分为预后良好组及预后不良组,并采用多因素Logistic回顾性分析法分析HR-HPV感染与食管鳞状细胞癌患者预后的关系。结果:384例食管鳞状细胞癌患者中HPV感染率为36.72%,其中低危HPV感染占比为9.93%,HR-HPV占比为90.07%,基因型由高到低分别为16型、18型、52型、33型、6型、51型、58型、11型;随访1年,384例食管鳞状细胞癌患者预后不良率为57.03%(219/384),预后不良组HR-HPV感染率明显高于预后良好组(P<0.05);经Logistic回顾分析显示,癌症低分化或未分化或中分化、临床分期Ⅳ期或Ⅲ期、淋巴结转移、饮酒史、HR-HPV感染均是食管鳞状细胞癌患者预后不良的危险因素(OR=3.916、2.581、4.080、3.238、4.821、2.986、3.062,P<0.05)。结论:合肥地区食管鳞状细胞癌患者HPV感染率较高,并以HPV16、18等高危型为主,且HR-HPV感染可增加食管鳞状细胞癌患者预后不良发生风险。     

人膀胱移行细胞癌HPV16/18型感染与c-erbB_2、H-ras、c-myc表达的关系

目的　研究人膀胱移行细胞癌 (TCC)中HPV 16 / 18型感染与c erbB2 、H ras、c myc蛋白产物表达的相互关系。方法应用免疫组织化学法检测经聚合酶链反应证实的 34例HPV 16 / 18感染阳性、2 0例HPV 16 / 18感染阴性的TCC组织和 7例正常膀胱组织中c erbB2 、H ras、c myc蛋白产物的表达 ,并经统计学处理。 结果　HPV 16 / 18感染阳性组c erbB2 、H ras、c myc蛋白产物表达阳性率分别为 5 5 .9% (19/ 34)、5 8.8% (2 0 / 34)、6 1.8% (2 1/ 34) ;HPV 16 / 18感染阴性组分别为 5 5 .0 % (11/ 2 0 )、6 5 .0 % (13/2 0 )、6 5 .0 % (13/ 2 0 ) ;正常膀胱粘膜上述 3种蛋白产物表达阳性例数依次为 0、1、0例。HPV 16 / 18感染与c erbB2 、H ras、c myc蛋白产物表达无关 (P >0 .0 5 )。c erbB2 、H ras、c myc蛋白产物阳性表达率在癌组织和正常膀胱粘膜之间有显著性差异 (P <0 .0 5 ) ,且其阳性表达率与TCC的病理分级相关 (P <0 .0 5 )。癌组织内c erbB2 与c myc蛋白产物表达呈正相关 (P <0 .0 1)。 结论　在TCC的发生、发展过程中 ,HPV 16 / 18可能不是主要通过c erbB2 、H ras、c myc蛋白产物的改变来发挥作用。c erbB2 、H ras、c myc蛋白产物的改变有可能为TCC发生的晚期事件 ,提示应注意其与临床预后的关系     

宫颈鳞状上皮病变组织中乳头状瘤病毒和Ki-67及p16INK4a蛋白表达意义的研究

目的:探讨Ki-67、p16INK4a和人乳头状瘤病毒(HPV)在不同程度宫颈鳞状上皮病变组织中的表达及临床病理意义.方法: 采用原位分子杂交及免疫组化方法,检测HPV的不同亚型、Ki-67、p16INK4a蛋白在182例不同程度宫颈病变组织中的表达.结果: HPV在不同程度病变中总检出率52.19%(95/182);在宫颈高级别上皮内瘤变及鳞癌组中检出最多的感染类型为HPV16/18,而HPV6/11在尖锐湿疣组检出率87.50%(21/24)最高;随着宫颈病变严重程度的增加,级别升高,Ki-67、p16INK4a阳性程度呈递增趋势.Ki-67、p16INK4a与HPV16/18型感染关系密切,χ2=11.779 8, P<0.01;Ki-67也与HPV6/11型有关.结论: HPV16/18型及Ki-67、p16INK4a 在宫颈高级别上皮内瘤变及鳞癌中表达明显升高,可能对宫颈鳞癌的发生、发展具有协同作用.     

食管鳞状细胞癌HPV 16/18型感染与p53表达的相关性研究

目的 探讨人乳头状瘤病毒（HPV）16/18型在食管鳞状细胞癌中的感染情况及其与p53的关系。方法 采用聚合酶链式反应（polymerase chain reaction,PCR）检测56例食管鳞状细胞癌手术切除组织（鳞癌组）和24名健康者正常食管组织（对照组）中HPV l6/18 E6、E7基因的表达,并根据其判断HPV感染情况;采用免疫组化SP法检测p53蛋白的表达水平。 结果 鳞癌组HPV感染率和p53蛋白阳性表达率均高于对照组（44.6% vs 12.5%,χ²=7.630,P=0.006;41.1% vs 4.2%,χ²=10.896,P<0.001）。在食管鳞癌组织中,HPV 16/18感染与病理分级、TNM分期和淋巴结转移有关（P<0.05）;且HPV16/18感染与p53蛋白表达呈正相关（r=0.565,P<0.001）。Kaplan-Meier生存分析结果显示,HPV感染阳性、p53表达阳性患者中位OS均小于HPV感染阴性、p53表达阴性者（21个月 vs 39个月,χ²=4.306,P=0.038;17个月 vs 41个月,^χ2=5.868, P=0.015）。控制相关的潜在混杂因素后,Cox回归模型显示HPV感染（HR=1.834,95%CI：1.010~3.330,P=0.046）和p53阳性表达（HR=2.189,95%CI：1.182~4.054, P=0.013）均可增加食管鳞状细胞癌患者的死亡风险。结论 食管鳞状细胞癌中HPV感染和p53阳性表达均较高,可能共同促进食管鳞状细胞癌的发生、发展并影响其预后。     

人乳头瘤病毒DNA在人喉癌细胞中表达的研究

为研究喉癌与人乳头状瘤病毒(HPV)感染的关系,本研究探讨了HPV在喉癌中的致病作用和基因组型的分布与表达。应用共有引物和多重引物PCR的方法,对160例喉不同病变的新鲜组织标本,进行HPV6、11、16、18、31、33、35、42、58共9型HPVsDNAs感染的检测。结果在喉癌组HPV感染的阳性率为49.3％(35/71),喉癌颈转移淋巴结组为22.7％(5/22),喉癌前病变组为11.8％(2/17),声带息肉组为6.7％(2/30),癌周正常喉组织为0％(0/20)。HPV DNA型别分布在喉癌中以HPV16、18型为主,喉良性病变中以HPV6、11型为主。表明喉癌发生发展与HPV感染相关。     

宫颈鳞状上皮病变组织中乳头状瘤病毒和Ki-67及p16^INK4a蛋白表达意义的研究

目的：探讨Ki-67、p16^INK4a和人乳头状瘤病毒（HPV）在不同程度宫颈鳞状上皮病变组织中的表达及临床病理意义。方法：采用原位分子杂交及免疫组化方法,检测HPV的不同亚型、Ki-67、p16^INK4a蛋白在182例不同程度宫颈病变组织中的表达。结果：HPV在不同程度病变中总检出率52.19%（95/182）;在宫颈高级别上皮内瘤变及鳞癌组中检出最多的感染类型为HPV16/18,而HPV6/11在尖锐湿疣组检出率87.50%（21/24）最高;随着宫颈病变严重程度的增加,级别升高,Ki-67、p16^INK4a阳性程度呈递增趋势。Ki-67、p16^INK4a与HPV16/18型感染关系密切,χ^2=11.779 8,P〈0.01;Ki-67也与HPV6/11型有关。结论：HPV16/18型及Ki-67、p16^INK4a在宫颈高级别上皮内瘤变及鳞癌中表达明显升高,可能对宫颈鳞癌的发生、发展具有协同作用。     

Two new human papillomavirus types (HPV54 and 55) characterized from genital tumours illustrate the plurality of genital HPVs

The genomes of two new human papillomavirus (HPV) types, named HPV54 and HPV55, were cloned from penile lesions of 2 patients. HPV54 was isolated from a verrucous carcinoma (Buschke-L?wenstein tumour) together with full-length HPV6 genomes and HPV6 DNA molecules with a deletion of about 0.3 kb located in the non-coding region. HPV55 was isolated from a condyloma acuminatum. No cross-hybridization was observed between HPV54 DNA and the DNAs of the known cutaneous and genital HPVs by blot hybridization experiments performed under stringent conditions. In contrast, significant cross-hybridization was detected between HPV55 DNA and the DNA of HPV13, associated with benign oral lesions, and, to a lesser extent, with the DNAs of HPV6, 11, and 44, associated with benign genital proliferative lesions. The DNA sequence homology between HPV55 and HPV6, 11, and 13 was estimated at 12%, 12%, and 20%, respectively, by hybridization in liquid phase at saturation, followed by nuclease S1 analysis. The physical maps of HPV54 and 55 were aligned with the genetic maps of HPV16 and 11, respectively, by heteroduplex mapping and partial DNA sequencing. HPV54 is thus only weakly related to the known HPVs, while HPV55 represents an additional HPV6-related HPV type. HPV54 and HPV55 are uncommon genital HPV types since, in a survey of a large series of specimens of benign, pre-malignant or malignant anogenital and orolaryngeal tumours, HPV54 was not detected, and HPV55 was found in another case of condyloma acuminatum.     

High-risk and multiple human papillomavirus infections associated with cervical abnormalities in Japanese women.

To estimate the risk of human papillomavirus (HPV) infection for cervical malignancies, we conducted a case-control study in Japan. Abnormal cervical cell (366) and normal cell samples (1562) were tested for the presence of HPV DNA using a new PCR-based test (LCR-E7 PCR). When single HPV infections were considered, 26 different HPV types were identified in normal cervices and in low-grade squamous intraepithelial lesions (LSIL); whereas HPV-16, -18, -31, -33, -35, -45, -51, -52, -56, -58 and -67 were detected in high-grade squamous intraepithelial lesions (HSIL) and in squamous cell carcinoma (SCC) of the cervix, and HPV-16 and -18 were detected in cervical adenocarcinoma. HPV-6 and -11 were detected in condyloma acuminatum tissue. In HSIL and SCC, HPV-16 was the most prevalent type and HPV-51, -52, and -58 were the next most prevalent; whereas HPV-39, -59, and -68 were not detected. Analysis by odds ratio (OR) revealed that HPV-11, -39, -42, -44, -53, -59, -62, and -66 (HPV-66: OR,139; 95% confidence interval (CI) = 6.7-168) were associated with LSIL; HPV-16, -18, -31, -51, -52 and -58 (HPV-16: OR, 69; 95%CI = 36-131) were associated with SCC; and HPV-16 and -18 (OR, 94; 95% CI = 28-317) were associated with adenocarcinoma. Multiple HPV infection was associated with LSIL (OR, 24; 95%CI = 13-44), HSIL (OR, 16; 95%CI = 8.4-32), and SCC (OR, 8.3; 95%CI = 3.2-22), although the prevalence decreased with the grade of the lesions. All results suggest that HPV-6 and -11 are condyloma types, HPV-16, -18, -31, -51, -52, -58, and perhaps -33, -35, -45, -56, and -67, are the high-risk HPV types, and many other types are LSIL-associated types in Japan. HPV typing and detection of multiple HPV infections in clinical samples may be useful as surrogate markers for cervical cell abnormalities.     

A Correlative Analysis of Cervical Lesions in Patients with Vulva Condyloma Acuminatum

T here are at least 40 subtypes of human papillomavirus (HPV) re- lated to infections and diseases of the female lower genital tract, including 13 definitely high-risk subtypes and 5 low-risk subtypes. In- fections by high-risk subtypes are associated wit…     

宫颈上皮内肿瘤(CIN)的克隆性状态与HPV感染的关系

目的:应用原位杂交方法检测不同克隆性状态的宫颈上皮内肿瘤(CIN)内人乳头状瘤病毒(HPV)感染率以及类型,探讨不同克隆性状态的CIN与HPV感染的关系,明确HPV(特别是高危型HPV)在单克隆性增生CIN和宫颈癌发生、发展中的作用。方法:采用原位杂交方法检测24例CINⅠ标本(2例单克隆性增生、22例多克隆性增生)、20例CINⅡ标本(13例单克隆性增生、7例多克隆性增生)、19例CINⅢ标本(均为单克隆性增生)HPV16/18、HPV6/11的表达状况。结果:34例单克隆性增生CIN中HPV16/18的阳性表达率为58.8%(20/34),HPV6/11的阳性表达率为26.5%(9/34),两者均为阳性5.9%(2/34),两者均为阴性20.6%(7/34);29例多克隆性增生CIN中HPV16/18的阳性表达率为10.3%(3/29),HPV6/11的阳性表达率为24.2%(7/29),两者均为阳性6.9%(2/29),两者均为阴性72.4%(21/29)。结论:单克隆性增生CINHPV16/18感染率明显高于多克隆性增生CIN(P<0.01)。HPV16/18感染是诱发单克隆性增生CIN重要因素。HPV16/18可能通过引起宫颈鳞状上皮的克隆性增生,促进细胞的转化和增殖,导致CIN的发生,并最终引起宫颈癌的发生、发展。     

Presence of human papilloma virus types 16 and 18 in genital warts and cervical neoplasias

Certain types of human papilloma viruses (HPV) are associated with human genital proliferative diseases, and among them HPV16 and HPV18 seem to play an important role in the occurrence of cervical cancer. We used restriction enzyme analysis and molecular hybridization, in order to investigate the type of viral infection and the physical state of viral DNA in gynecological benign, pre-malignant and malignant lesions. HPV6/11 specific sequences could only be detected as episomes and this in benign lesions classified as condylomata acuminata. On the other hand, HPV16 and HPV18 sequences were detected in non-malignant lesions such as flat condylomata (7 out of 14 cases), pre-malignant lesions including cervical intra-epithelial neoplasias (10 out of 20 cases), and most frequently in cervical invasive cancers (21 out of 27 cases). In a large number of virus-positive cases, HPV16 and HPV18 could only be discerned in forms consistent with the existence of episomes and/or randomly integrated head-to-tail oligomers. However, some invasive carcinomas and cervical intra-epithelial neoplasias contained, in addition, clonal outgrowths with detectable virus-cellular junction fragments of the integrated viral genomes. In the light of these data, monitoring the type of viral infection proves to be an important adjunct to histological analysis when assessing those patients affected by condyloma or cervical intra-epithelial neoplasia who are at risk for developing invasive carcinoma.     

Cloacogenic carcinoma of the anal canal and associated viral lesions. An in situ hybridization study for human papilloma virus.

Five cases of cloacogenic carcinoma were analyzed for microscopic human papilloma virus (HPV)-induced changes and with in situ hybridization technique for HPV types 6/11, 16/18 and 31/35/51. Four of the five cases showed epithelial foci of koilocytotic atypia. HPV type 16/18 was present in four of the five cases. The surface epithelium in two of the four cases with koilocytotic changes showed HPV type 16/18. HPV type 6/11 was seen in surface epithelium in one case in which invasive carcinoma showed HPV type 16/18. This double infection with double morphologic expression could mean that the same behavioral pattern (anal intercourse) may contribute to both anal condyloma and carcinoma and, although patients with condyloma are at risk for carcinoma, condyloma may not be the precursor lesion in all cases with coexistent condyloma and carcinoma. Because of the similarity between pathogenesis of anal and cervical carcinomas, a periodic cytologic screening of anal mucosa could be indicated in populations at risk: homosexual men, patients with condyloma, women having dysplasia or carcinoma of the uterine cervix, and patients with immunosuppressive disorders.     

Occurrence of human papillomavirus DNA in primary lung neoplasms.

The occurrence of human papillomavirus (HPV) DNA in primary lung carcinomas and in squamous metaplasia of the bronchus was studied using in situ hybridization techniques and commercially available biotinylated DNA probes to HPV subtypes 6/11, 16/18, and 31/33/35. The authors found HPV DNA in six of 20 cases of squamous cell carcinoma and one of six cases of large cell undifferentiated carcinoma. There were two cases each of the 6/11 serotypes and the 16/18 serotypes and three cases of the 31/33/35 serotypes. Infected cells of the squamous carcinomas uniformly showed koilocytosis. No case of adenocarcinoma, bronchioloalveolar carcinoma, or small cell carcinoma was positive (of 32 cases). Areas of squamous metaplasia in infected tumors showed similar HPV DNA expression in 15% of cases, especially in those with condylomatous atypia. In 5.8% of random bronchial biopsies of squamous metaplasia, HPV DNA was identified. The relationship of HPV infection to the development of upper and lower respiratory tract carcinomas is discussed.     

Histological types of carcinoma of the uterine cervix and the detectability of human papillomavirus DNA

Using the Southern DNA hybridization technique, tissues from 17 cases of invasive carcinoma of the uterine cervix, including nine cases of squamous cell carcinoma, four cases of adenocarcinoma, one case of adenosquamous carcinoma, and three cases of undifferentiated carcinoma, were examined for the presence of human papillomavirus (HPV) DNA. None of the studied cases had histologically confirmed association of condyloma acuminatum or cervical intraepithelial neoplasia in the vicinity. HPV DNA was detected in two of 17 cases under low stringency conditions. One lesion was undifferentiated carcinoma, and another was squamous cell carcinoma. Hybridization under high stringency conditions with a variety of HPV DNA probes indicated the presence of HPV-16 in these two lesions. The other HPV-positive lesion was adenocarcinoma, demonstrating weak hybridizations with HPV-2 and HPV-16 DNA probes only under high stringency conditions. Altogether, three of 17 cases (17.6%) contained HPV DNA. This observation contrasts to the rate of HPV DNA present in 15 of 18 cases (83.3%) of the tissues of cervical intraepithelial neoplasia. Our data suggest that HPV was not consistently detected in invasive squamous cell carcinoma, despite the frequent association of HPV with its supposed precursor lesions of cervical intraepithelial neoplasia.