鲍恩样丘疹病皮损中HPV16、18的检测

目的：探讨人乳头瘤病毒（HPV）16、18在鲍恩样丘疹病皮损中的感染情况。方法：采用杂交捕获试验筛选高危型HPV和低危型HPV阳性标本,对高危型HPV阳性标本经PCR技术检测其HPV16、18DNA阳性者。结果：45例鲍恩样丘疹病患者皮损中34（75.6%）例为高危型HPV阳性,在高危型HPV阳性标本中82.2%（28/34）为HPV16阳性,5.9%（2/34）为HPV18阳性,8.8%（3/34）HPV16、18均阳性,2.9%（1/34）未检测到HPV16及HPV18DNA。结论：多数鲍恩样丘疹病皮损中存在高危型HPV感染,尤其HPV16,故高危型HPV16感染与鲍恩样丘疹病的发生、发展密切相关。     

利用重组HPV16L1蛋白检测鲍温病和鲍温样丘疹病血清中的抗体

目的用原核表达系统获得人乳头瘤病毒16型(HPV16)主要衣壳蛋白(L1),检测鲍温病和鲍温样丘疹病血清中的HPV16L1IgG抗体。方法PCR方法检测鲍温病和鲍温样丘疹病局部皮损中的HPVDNA,限制性片段长度多肽性方法进行HPV分型;用原核表达载体pET32a表达HPV16L1蛋白,亲和层析法对表达产物进行纯化,获得了HPV16L1蛋白抗原;ELISA法检测鲍温病和鲍温样丘疹病血清中的抗体。结果用原核表达系统和镍柱亲和层析获得了较纯的HPV16L1蛋白。鲍温样丘疹病血清中抗体的检测率为(20/50)40%,鲍温病血清中抗体的检测率为(8/30)26.7%,正常献血员组检测率为(2/50)4%。这两组分别与正常献血员组比较有统计学差异(P0.05)。两组HPV16DNAPCR检测法与血清HPV16L1抗体检测法有较好的一致性(P0.05)。结论利用原核表达系统获得纯化HPV16型L1蛋白,可用于鲍温样丘疹病和鲍温病血清中HPV16抗体的检出,为其临床上血清学诊断提供参考,进一步开发HPV相关皮肤恶性肿瘤高危人群的诊断试剂盒提供前沿基础研究。     

14例鲍温样丘疹病皮损人类乳头瘤病毒DNA原位杂交的检测

应用地高辛标记的 HPV6B/11、16、18型 DNA探针对 14例鲍温样丘疹病进行原位杂交检测。结果显示 14例中 7例 HPV 6B/11和 18型阳性 ,两者反应强度相似 ,即强阳性 3例 ,弱阳性 4例 ,而6B/11、18型阳性同时伴 16型阳性的仅 3例 ,且 16型反应较弱 ,表明有半数鲍温样丘疹病的发病与 6B/11、18型 HPV感染有关     

原位杂交检测鲍温样丘疹病及Bowen病中HPV16 DNA

采用生物素标记的核酸探针原位杂交技术,检测了30例鲍温样丘疹病、15例Bowen病中16型人乳头瘤病毒(HPV16)感染的组织定位及染色模式。结果显示,30例鲍温样丘疹病中,HPV16阳性13例;15例Bowen病中,HPV16阳性6例。鲍温样丘疹病、Bowen病中HPV16感染累及棘细胞全层;主要为核内团块状着色。Bowen病中,HPV16感染尚累及基底层细胞且存在HPV的点状染色模式。提示鲍温样丘疹病与Bowen病比较,HPV16感染的组织定位及染色模式均有所不同。     

细胞周期蛋白E和p27在鲍温样丘疹病和外生殖器表皮原位癌中的表达

目的：探讨细胞周期相关因子细胞周期蛋白E及其依赖性蛋白激酶抑制蛋白p27，在高危型人乳头瘤病毒(HPV)相关的鲍温样丘疹病及外生殖器表皮原位癌中的变化及意义。方法：采用通用型引物聚合酶链反应-限制性片段长度多态性(PCR—RFLP)方法对51例外生殖器部位上述病变进行HPVDNA的检测和分型，应用免疫组化方法检测病变中细胞周期蛋白E和p27的表达。结果：40例鲍温样丘疹病、5例生殖器鲍温病及6例Queyrat增殖性红斑HPVDNA的阳性率分别为55．0％、100．0％和33．3％，HPV16为主要型别；高危型HPV相关的病变中细胞周期蛋白E的表达明显高于正常上皮，且HPV阳性组明显高于HPV阴性组，而p27表达明显低于正常上皮，且HPV阳性组明显低于HPV阴性组。上述疾病中细胞周期蛋白E与p27的表达呈负相关。结论：HPV16感染与鲍温样丘疹病及外生殖器表皮原位癌的发生有密切关系，并可能通过影响细胞周期蛋白E和p27表达及两者的相互作用引起感染上皮的增生及恶变。     

尖锐湿疣及鲍恩样丘疹病皮损中高危型人乳头瘤病毒感染与端粒酶表达的关系

目的:研究尖锐湿疣(CA)与鲍恩样丘疹病(BP)皮损中高危型人乳头瘤病毒(HPV)感染与端粒酶表达的关系。方法:采用PCR方法检测HPV类型,免疫组化方法检测CA和BP皮损中人类端粒酶反转录酶(hTERT)的表达。结果:CA和BP皮损中hTERT表达均比正常对照组高(P均<0.01),BP又较CA表达高(P<0.01)。CAHPV16阳性组hTERT表达较HPV16阴性组高,其差异有统计学意义(P<0.05);BP不同HPV型之间hTERT表达的结果与CA相同;以上各组均比正常对照组表达强度高(P<0.05)。结论:非恶性增殖性疾病中亦有端粒酶不同程度的表达;以HPV16为代表的高危型HPV感染与端粒酶表达有相关性。     

病毒性皮肤病

980603 14例鲍温样丘疹病皮损人类乳头瘤病毒DNA原位杂交的检测/纪华安（天津市长征医院病理室）…//中国皮肤性病学杂志.-1997,11（5）.-266 为探讨人类乳头瘤病毒（HPV）在鲍温样丘疹病（BP）皮损中的表达特点,应用地高辛标记的HPV 6B/11、16、、18型DNA探针,对14例BP患者皮肤和粘膜病理组织进行了原位杂交检测。结果:BP患者HPV6B/11阳性7例（50.0％）,其中强阳性3例     

外生殖器部位皮肤肿瘤临床病理和HPV感染的相关性

目的:探讨外生殖器部位皮肤肿瘤和人乳头瘤病毒(human papilloma viruses,HPV)感染的相关性。方法:采用通用型引物-PCR方法对89例外生殖器部位皮肤肿瘤进行HPV DNA的检测,应用限制性内切酶酶切片段长度多态性(RFLP)方法进行HPV DNA分型。结果:40例鲍温样丘疹病中22例(55%)HPV DNA阳性,其中20例为HPV16,1例为HPV31,1例为HPV6+16;5例鲍温病HPV DNA全部阳性(100%),其中4例为HPV16,1例为HPV6+16;6例Queyrat增殖性红斑中2例(33.3%)HPV DNA阳性,均为HPV16。18例外生殖器鳞状细胞癌中5例(27.8%)HPV DNA阳性,均为HPV16;20例乳房外Paget病HPV DNA为阴性。结论:外生殖器部位皮肤肿瘤中鲍温样丘疹病、鲍温病、Qu-eyrat增殖性红斑的发生可能与HPV16感染有密切关系;外生殖器鳞状细胞癌中,HPV16感染可能是多种致癌因素中的一个重要因素;外生殖器部位乳房外Paget病的发病可能与HPV感染无关。     

鲍恩样丘疹病HPV感染型别与组织病理及临床表现的相关性

目的:探讨鲍恩样丘疹病HPV感染型别与病理组织图像、临床表现的相关性。方法:通过基因芯片技术对44例鲍恩样丘疹病患者的病理样本进行HPV检测及分型,根据HPV高危程度以及是否含HPV16型分为低危混合组(A组,6例)、不含HPV16型高危混合组(B组,8例)、含HPV16型高危混合组(C组,30例),分别观察A、B、C 3组相应的组织病理图像及临床表现。结果:(1)44例患者的病理标本HPV DNA检测均为阳性,检出总亚型数为76例次,低危型占36.8%,高危型占63.2%。(2)A、B、C3组患者病理组织图像均有不同程度的不典型增生改变,不典型增生程度与HPV低高危致癌性基本一致。结论:鲍恩样丘疹病患者HPV感染高危型以HPV16型为主,低危型以HPV6为主。HPV型别与组织病理图像、临床表现有一定的相关性,该研究为通过组织病理图像结合临床表现反推HPV感染的可能型别研究打下基础。     

女性鲍恩样丘疹病患者外阴皮损及宫颈HPV感染情况

近年研究发现,鲍恩样丘疹病的发生与人乳头瘤病毒(HPV)感染有关,尤其是高危型HPV。高危型HPV与宫颈癌的发生密切相关。为探讨女性鲍恩样丘疹病外阴皮损及宫颈HPV的感染情况,我们对18例女性鲍恩样丘疹病患者的外阴皮损及宫颈进行了低危型HPV6/11及高危型HPV16、18DNA检测。     

外阴鳞状细胞癌中人乳头瘤病毒感染与端粒酶和生存素的关系

目的:探讨人乳头瘤病毒(HPV)6/11、16/18在外阴鳞状细胞癌(vulvarsquamouscellcarcinoma,VSCC)组织中的感染情况及其与人端粒酶逆转录酶(hTERT)和凋亡抑制基因生存素(survivin)表达的关系。方法:采用PCR检测HPV6/11、16/18在31例VSCC及13名正常人皮肤组织中的感染情况,同时用原位杂交法检测hTERTmRNA的表达,免疫组化法检测生存素蛋白的表达。结果:①HPV6/11、16/18在VSCC患者的阳性率分别为25.81%和38.17%,正常对照者为阴性。VSCC患者与正常对照者HPV16/18阳性率的差异有统计学意义(P<0.05)。②VSCC患者hTERTmRNA、生存素蛋白表达的阳性率与正常对照者比较,差异均有统计学意义(P<0.05),且VSCC患者hTERTmRNA表达与生存素蛋白表达的阳性率呈明显的正相关(P<0.05)。③hTERTmRNA在HPV16/18阳性组中的表达明显强于HPV16/18阴性组,生存素蛋白在HPV16/18阳性组中的表达低于其在HPV16/18阴性组中的表达。结论:HPV感染及hTERT、生存素表达在VSCC发生发展中起一定作用;VSCC中hTERT与生存素的表达具有一定的相关性。     

原位分子杂交检测尖锐湿疣组织中HPV DNA的研究

本文采用HPV DNA原位分子杂交法检测了30例尖锐湿疣组织标本，结果显示HPV6/11型阳性检出率为86.6%，HPV16/18型为16.6%，HPV31/33/35则为阴性，总的阳性检出率为90%。阳性结果作见于表皮浅层外，还见于棘细胞层中下部及基底细胞层内，作者认为原位分子杂交法是目前检测HPV感染及其分型的一种敏感、特异、快速且相对简便的方法，又能进行感染的组织学定位，同时这一方法还适应于对以往病例进行回顾性调查。     

Detection of human papillomavirus in cutaneous extragenital Bowen's disease in immunocompetent patients

INTRODUCTION: A specific link between human papillomavirus (HPV) types 16, 18, 31, and 33 and genital carcinomas and between HPV type 5 and cutaneous extragenital carcinomas in patients with epidermodysplasia verruciformis and renal transplant has been previously found. The aim of this prospective study was to detect HPV in cases of cutaneous extragenital Bowen's disease (BD) from non-immunosuppressed patients. PATIENTS AND METHODS: Twelve cases of cutaneous extragenital BD or Bowen's carcinoma (BC), seen in the period 1994-1996 and confirmed by histologic examination, were included in the study. Tissue sections were studied by in situ hybridization with a mixture of HPV DNA probes and specific HPV DNA probes. In addition, study on fresh materiel from 1995 included: Southern blot hybridization with various usual HPV probes (6, 11, 16, 18, 31, 33, 35, 39, 42), polymerase chain reaction (PCR) with hybridization using consensus HPV probes and probes specific for HPV types 6, 11, 16, 18 and 33. In positive samples with conventional PCR, in situ PCR with probes specific for HPV types 6/11 and 16 was performed on tissue sections. RESULTS: In situ hybridization was negative in all the cases. Southern blot hybridization was negative in our 9 studied cases. Three cases studied by consensus PCR were positive. PCR with specific HPV probes revealed positivity on two of these cases: HPV 6 in one, and HPV 16 in another. In situ PCR was positive with a mixed 6/11 HPV probe in the third positive consensus PCR case. DISCUSSION: Our study revealed the presence of HPV in 3 out of 12 cases of cutaneous extragenital BD and BC. HPV type 16, found in BC of skull, was the most usually found type in the literature. HPV types 6/11, detected in 2 cases, were rarely found in cutaneous extragenital BD and BC and these results are in favor of the oncogenic effect of these virus types. In our study, in situ hybridization and Southern blot hybridization were negative in all the cases; HPV was only found in 3 cases by conventional PCR and in 1 case by in situ PCR. The low range of detection of HPV in cutaneous extragenital BD may be due to the used methods, to difficulties related to sampling and/or to a low number of copies of the HPV genoma.     

Simultaneously detected aberrant p53 tumor suppressor protein and HPV-DNA localize mostly in separate keratinocytes in anogenital and common warts

Abstract The E6 oncoprotein of human papillomavirus (HPV) is known to inactivate the control function on cell cycle exerted by p53 tumor suppressor protein in vitro by binding to p53 and thus facilitating the degradation of p53. We have applied a simultaneous in situ demonstration method for detecting p53 protein and HPV-DNA on formalin-fixed tissue sections, and investigated the in vivo interrelationship of p53 protein and HPV-DNA. Immunohistochemical staining for p53 protein with polyclonal and monoclonal antibodies, recognizing both wild-type (wt) and mutated p53 protein, was performed first and in situ DNA hybridization (ISH) for HPV types 6/11 or 16/18 with digoxigenin-labelled probes thereafter. 47% (25/53) of 48 histologically confirmed primary or recurrent condylomata acuminata (CA), 2 Bowenoid papulosis (BP) and 3 common wart (CW) biopsies, positive for HPV 6/11 or HPV 16/18 DNA, showed keratinocytes immunopositive for p53 protein. Of these. 11 lesions with abundant numbers of p53-positive cells were further analyzed with the double method. Signals for abnormal p53 protein and HPV-DNA were detected in separate cell nuclei in all biopsies and, additionally, in the same cell nuclei in 3 biopsies (1 BP, 1 CA, 1 CW). Usually the p53 positivity localized more basally in the epidermis than HPV-DNA, although p53- and HPV-positive keratinocytes were always located closely. The findings were similar for HPV-types 6/11 and 16/18. Our finding of both p53 and HPV-6/11 signals in the same cell nuclei may indicate complexing of p53 and low-risk HPV's without degradation of p53. Our results show abnormal p53 expression in HPV-infected skin lesions, and suggest that p53 protein is susceptible to aberrations even in the cells in the vicinity of productive HPV infection. However, it is not yet fully understood how HPV interferes with p53 protein in these cells.     

Oncogenic human papillomaviruses in extra-genital Bowen disease revealed by in situ hybridization]

BACKGROUND: The association between mucosal oncogenic human papillomaviruses (HPV) and bowenoid papulosis or genital Bowen's disease is well documented. In contrast this association with extra-genital Bowen's disease is poorly studied. The aim of this study was to detect oncogenic (16/18, 31/33/51) and non oncogenic (8/11) mucosal HPV using a in situ hybridization method in 28 skin biopsy specimens of extra-genital Bowen's disease.PATIENTS AND METHODS: Twenty-eight cases of extra-genital Bowen's disease seen in the period 1990-96 in the Dermatology department were included: 19 women and 9 men (mean age: 72 years). Bowen's disease locations were: hands and feet (8 cases), limbs (11 cases), face (8 cases), trunk (1 case). Blinded histopathologic examination confirmed the diagnosis of Bowen's disease and signs of HPV infection (koilocytosis). In situ hybridization was performed using three biotinylated probes detecting HPV types 6/11, 16/18, 31/33/51.RESULTS: Oncogenic HPV genoma was detected in 8 skin samples (28.6 p. 100). In all these cases, 16/18 probe was positive and in two cases, both 16/18 and 31/33/51 probes were positive; 4/8 Bowen's diseases of the extremities were positive for HPV. Koilocytes were found in 6/8 of skin samples with positive HPV detection.DISCUSSION: Mucosal oncogenic HPV are detected by in situ hybridization in 28.6 p. 100 of extra-genital Bowen's disease. In situ hybridization is an easier technique than Southern-Blot hybridization which is the gold standard. Five studies reported similar results and three studies reported different results that we discuss. A precise understanding of oncogenic HPV implication in the development of extra-genital Bowen's disease could lead to the development of new therapeutic strategies (topical cidofovir or imiquimod).     

外阴鳞状细胞癌中人乳头状瘤病毒感染与基质金属蛋白酶9和金属蛋白酶抑制剂1的关系

目的 探讨人乳头状瘤病毒(HPV)6/11、16/18在外阴鳞状细胞癌(简称外阴鳞癌)组织中的感染情况及与基质金属蛋白酶9(MMP-9)和金属蛋白酶抑制剂1(TIMP-1)蛋白表达的关系.方法 用原位杂交法检测HPV6/11、16/18在26例外阴鳞癌、21例外阴上皮内瘤病变(VIN)及10例外阴正常皮肤组织中的感染情况.同时用免疫组化法检测MMP-9、TIMP-1蛋白的表达.结果 HPV6/11、16/18在外阴鳞癌、VIN中的感染率分别为69.23%(18/26)和38.46%(10/26),42.86%(9/21)和28.57%(6/21),正常对照组没有感染.MMP-9、TIMP-1蛋白在外阴鳞癌、VIN、正常对照组中的阳性表达率分别为92.31%(24/26)和76.92%(20/26),90.48%(19/21)和80.95%(17/21),80.00%(8/10)和50.00%(5/10).HPV6/11的外阴鳞癌组、VIN组与正常对照组比较差异有显著性(P<0.05),HPV16/18的外阴鳞癌组与正常对照组比较差异有显著性.外阴病变各组MMP-9、TIMP-1阳性表达率与正常对照组比较差异均有显著性.在外阴鳞癌中MMP-9蛋白表达较TIMP-1蛋白表达强(P<0.05).MMP-9在有HPV6/11和HPV16/18感染患者中的表达较无感染者强,而TIMP-1蛋白的表达差异无显著性.结论 在外阴鳞癌的发生发展中HPV的感染起一定作用,当MMP-9的表达强于TIMP-1表达时可能是外阴鳞癌侵袭的一个指标,HPV感染可能会进     

MDM2蛋白和MDM2 mRNA在尖锐湿疣中的表达

目的 研究尖锐湿疣中MDM2蛋白及MDM2 mRNA的表达情况,探讨MDM2在尖锐湿疣增殖及癌变中的作用。方法 分别采用免疫组化染色及原位杂交方法检测外阴尖锐湿疣和正常人皮肤组织中MDM2蛋白及mRNA的表达情况,同时用PCR法检测人乳头瘤病毒(HPV)型别。结果 32例外阴尖锐湿疣中MDM2蛋白阳性表达18例(56.25%),MDM2 mRNA阳性表达22例(68.75%)。MDM2蛋白与mRNA共同阳性表达14例。PCR检测示尖锐湿疣皮损中HPV6/11型28例(87.5%),HPV16/18型4例(12.5%)。在18例MDM2蛋白阳性表达的标本中,HPV6/11型15例,HPV16/18型3例。在22例MDM2 mRNA阳性表达的标本中,HPV6/11型18例,HPV16/18型4例。而正常人皮肤中MDM2蛋白及MDM2 mRNA均未见表达。结论 MDM2的异常表达可能与尖锐湿疣过度增殖及癌变有关。     

尖锐湿疣人乳头瘤病毒的基因分型

目的：检测尖锐湿疣中人乳头瘤病毒（HPV）的基因型别。方法：用通用引物进行聚合酶链反应,扩增尖锐湿疣中HPV DNA,用生物素标记的特异性基因分型探针,对聚合酶链反应扩增后的HPV DNA进行斑点杂交,结果：50例尖锐湿疣标本经PCR扩增后48例HPV DNA阳性,阳性标本中HPV6,11,16,18型的检出率分别为16．7％（8／48）,37．5％（18／48％）,14．6％（7／48）,4．2％（2／48）,多重型别检出率为22．9％（11／48）,结论 HPV11型感染是尖锐湿疣发病的首要原因。     

Melanocytic proliferation in condylomata acuminata. A report of two cases and investigation by in situ hybridisation

Two cases of melanocytic lesion occurring in condylomata acuminata are described. In situ hybridization with human papillomavirus (HPV) DNA probes specific for 6b, 11, 16, 18 revealed positivity with HPV6 and 11, in the non-dysplastic condylomata, and HPV 18 positivity in the case showing severe dysplasia. The HPV localization was confined to the superficial parakeratotic zones, remote from the melanocytic proliferation. The relationship between human papillomavirus and the melanocytic lesions is discussed.