鲍温样丘疹病中hTERT蛋白表达与高危型HPV感染的相关性研究

目的研究hTERT蛋白在鲍温样丘疹病中的表达及与高危型HPV存在的相关性。方法通过原位杂交法研究HPV的存在和免疫组化法检测hTERT蛋白在鲍温样丘疹病中的表达,并探讨hTERT蛋白表达与高危型HPV存在的相关性。结果在26例鲍温样丘疹病标本中,18例(69.2%)为HPV16/18阳性,其中1例HPV16/18阳性标本也有HPV6/11阳性。3例(11.5%)为HPV31/33/35阳性,2例(7.7%)为未定型HPV阳性,21例有高危型HPV存在的标本是hTERT阳性,16例高危型HPV感染的标本呈现了强而弥漫性hTERT蛋白表达。hTERT蛋白表达与高危型HPV的存在显著相关。结论高危型HPV可能诱导了hTERT蛋白的高表达。     

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     

某些外生殖器表皮肿瘤皮损中HPVDNA的检测和分型

目的探讨人乳头瘤病毒(HPV)感染与外生殖器部位表皮增生性疾病发生的相关性。方法采用通用型引物PCR方法对151例外生殖器表皮增殖性病变进行HPVDNA的检测,应用限制性内切酶片段长度多态性(RFLP)方法进行HPVDNA分型。结果30例尖锐湿疣HPVDNA全部阳性,其中HPV6占60%,HPV11占26.7%;40例Bowen样丘疹病、5例生殖器部位Bowen病、6例Queyrat增生性红斑HPVDNA的阳性率分别为55%、100%、33.3%,主要型别为HPV16;18例外生殖器浸润性鳞状细胞癌中HPVDNA的阳性率为27.8%,均为HPV16;32例黏膜白斑和20例乳房外Paget病HPVDNA均为阴性。结论尖锐湿疣主要由HPV6,其次由HPV11感染所致;Bowen样丘疹病、表皮原位癌包括Bowen病、Queyrat增生性红斑的发生可能与HPV16感染有密切关系;外生殖器浸润性鳞状细胞癌中,HPV16感染可能是多种致癌因素中的一个重要因素;外生殖器部位黏膜白斑和乳房外Paget病的发病可能与HPV感染无关。     

鲍温样丘疹病的诊断与治疗

鲍温样丘疹病(bowenoid papulosis,BP)是由1970年Lloyd首先报道,并命名为多中心性色素性鲍温病(multicentric pigmented Bowen’s disease),之后相继有文献报道,其名称有:生殖器多中心鲍温病、鲍温样非典型增生、可逆性女阴异型和原位变化的色素性丘疹病等。1978年Wade等人将其命名为“鲍温样丘疹病”,并延用至今。但此病作为一种具有独立临床、病理特征的疾病越来越引起人们的重视,对此病的研究和相关报道也是逐年增多。1病因和发病机制绝大部分学者认为BP的发病与HPV(human pap-illoma virus)感染有关,尤其是HPV16,病毒学研究认为是…     

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。     

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。     

鲍温样丘疹病的临床及感染人乳头瘤病毒类型的研究

目的 研究鲍温样丘疹病(BP)的诊治及其感染人乳头瘤病毒(HPV)的类型。方法 对9例BP的临床表现、组织病理及治疗进行总结,用PCR法检测13例BP石蜡组织标本HPV16 L1基因。结果 BP表现为生殖器部位的褐色扁平丘疹,有线状排列趋势。病理表现为角化不全,棘层肥厚,角质形成细胞出现异型性并排列如伏麦状,基底膜完整。8例经疣体破坏、干扰素及免疫增强剂治疗,全部治愈;1例行手术局部扩大切除后复发,再经上述方案治疗而愈13例 BP标本有12例 HPV阳性,都为HPV16。结论 BP主要由HPV16感染引起,临床和病理表现特异,疣体玻坏结合干扰素及免疫增强剂治疗,疗效较好。     

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。     

鲍温样丘疹病35例临床分析

目的分析鲍温样丘疹病临床诊断治疗中的特点与意义。方法回顾分析我院近10年来35例鲍温样丘疹病患者的临床资料。结果（1）各种合并症约占鲍温样丘疹病总数的80％；（2）误诊者约占鲍温样丘疹病总数的71％。结论（1）鲍温样丘疹病与HPV感染有密切的关系；（2）本病鉴别诊断上需要病理与临床相结合；（3）免疫力下降与本病有关；（4）对于顽固性鲍温样丘疹病加以配合口服中药治疗，可取得良好效果。     

鲍温样丘疹病患者人类乳头瘤病毒分型及疗效分析

目的探讨鲍温样丘疹病(BP)皮损中人乳头瘤病毒(HPV)感染类型与患者疗效的关系。方法用基因芯片法检测BP皮损组织中HPV感染类型,再用艾拉光动力和二氧化碳激光两种不同手段治疗鲍温样丘疹病患者并进行对照。结果92例BP中HPV高危型感染48例,占52.17%;高低危混合感染者28例,占30.43%;低危感染16例,占17.4%;采用不同的治疗方法治疗表明,艾拉光动力是治疗鲍温样丘疹病的有效方法,总有效率达92.75%,其中治愈率为85.5%。结论 BP的发生与发展和HPV感染密切相关。治疗效果与病毒感染类型无直接关系。采用二氧化碳激光加艾拉光动力治疗BP疗效优异,副作用小,是值得推广的治疗方法。     

Detection of mucosal human papilloma virus DNA in bowenoid papulosis, Bowen's disease and squamous cell carcinoma of the skin

Human papilloma virus (HPV) is known to be an etiologic agent for benign warts of the skin. Recently, HPV have been detected in malignant skin and mucosal diseases suggesting that HPV infection can induce malignant skin tumors. In the present study, we examined the presence of mucosal HPV DNA in normal tissue, Bowen's disease (BD), Bowenoid papulosis (BP) and squamous cell carcinoma (SCC) of the skin. We detected the HPV DNA with polymerase chain reactions, and identified the type by DNA sequencing. In the results, we detected HPV DNA in none of the 17 normal controls, two of the three BP (66.7%), one of the 21 BD (4.8%), and six of the 26 SCC of the skin samples (23.0%). The occurrence rates of HPV in BP and SCC were significantly elevated compared to that of normal controls (P < 0.01 and P < 0.01, respectively). In addition, the occurrence rate of HPV in BP was significantly elevated compared to that of BD (P < 0.05). The reproducibility was confirmed with a polymerase chain reaction (PCR) with another primer pair. Of the two cases of BP with positive HPV DNA, one case showed HPV 31 and the other case HPV 16. The case of BD with positive HPV DNA showed HPV 31. Of the six cases of SCC with positive HPV DNA, one case showed HPV 16, another case HPV 34, and the other four cases HPV 31. These results showed that mucosal HPV, including HPV 31 and 16, could be detected in SSC of the skin. Mucosal HPV, not only the epidermodysplasia verruciformis type, appear to induce malignant skin tumors.     

尖锐湿疣、鲍温样丘疹病及鲍温病皮损中人乳头瘤病毒感染与P16蛋白表达的关系

目的：探讨鲍温样丘疹病（BP）、鲍温病（BD）及尖锐湿疣（CA）皮损中高、低危人乳头瘤病毒（HPV）感染对P16蛋白表达的影响。方法：用荧光定量PCR（FQ－PCR）方法检测30例CA病损及12例正常人皮肤粘膜中HPV6／11、HPV16／18DNA；用原位杂交方法检测30例BP、15例BD中HPV16DNA；用免疫组化方法检测上述组织中P16蛋白、Ki-67蛋白的表达。结果：在HPB感染皮损中，P16蛋白、Ki-67蛋白的表达均增高（P＜0．05），且在HPV16阳性的BP、BD中的表达高于HPV6／11感染的CA（P＜0．05）；P16蛋白表达与Ki-67蛋白表达呈正相关（P＜0．05）。结论：HPV感染皮损中，P16蛋白的过度表达可能是机体对HPV感染引起细胞异常增殖的一种反馈调节；P16蛋白表达的不同可能是由于高、低危HPV引起的Rb蛋白功能失活不同。     

Human papillomavirus infection in Bowen disease: Negative p53 expression,not p16INK4a overexpression,is correlated with human papillomavirus‐associated Bowen disease

Genital Bowen disease (BD) has been linked to the high‐risk types of human papillomavirus (HPV) infection. Recently, it has been recognized that HPV also can be associated with extragenital BD. HPV oncoproteins E6 and E7 interfere with the function of p53 and pRb, respectively, leading carcinogenesis. p16^INK4a overexpression induced by inactivation of pRb is recognized as a surrogate marker for HPV‐associated cervical cancer. In this study, we examined the presence of HPV DNA in 142 BD lesions by polymerase chain reaction (PCR), and determined the type of HPV by PCR restriction fragment length polymorphism or direct DNA sequencing. HPV DNA was detected in 66.7% of genital BD and 8.3% of extragenital BD. The types of HPV detected were HPV types 6, 16, 33, 52, 56, 58 and 59. We also investigated the expression of p16^INK4a, pRb and p53 by immunohistochemistry. Positive expression was detected in 88.6% for p16^INK4a, 25.2% for pRb, and 63.8% for p53. There was no significant difference in p16^INK4a and pRb expression between HPV‐positive and ‐negative BD. However, a strong correlation of HPV positivity with p53 negativity was found. A total of 66.7% of HPV‐positive BD showed no p53 expression, whereas the corresponding rate was 32.8% of HPV‐negative BD. This study demonstrated that HPV can participate in the development of BD, not only in the genital lesion, but also in extragenital lesion. p16^INK4a overexpression is not a marker for HPV infection in BD. Instead, negative p53 expression is correlated with HPV‐associated BD.     

聚合酶链反应及转印杂交对尖锐湿疣复发的病因学研究

用通用引物介导的聚合酶链反应及其产物斑点杂交技术，对１５例复发及相应的原发尖锐湿疣（ＣＡ）皮损中人乳头瘤病毒相关序列进行了检测和分型；同时还对３５例原发ＣＡ周围外观正常皮肤组织ＤＮＡ作了转印杂交分析。结果表明隐性感染是ＣＡ复发的非常重要原因，再感染也是ＣＡ复发不容忽视的原因。     

Human papillomavirus-32-positive extragenital Bowenoid papulosis (BP) in a HIV patient with typical genital BP localization

OBJECTIVES: Bowenoid papulosis (BP) is characterized by multiple maculopapular lesions of the genitalia; extragenital localization is rarely found. Human papillomavirus (HPV) type 16 has been found in most cases of BP, but the other HPV genotypes associated with BP had been poorly characterized. GOAL: We describe an extragenital BP with concomitant genital involvement in an HIV-positive woman. STUDY: Several HPV gene-specific amplifications and sequencing were performed on DNA extracted from biopsy samples. RESULTS: The presence of HPV-16 DNA in anogenital and HPV-32 DNA in lip lesions was demonstrated in the absence of any other HPV type and with no coinfection. Semiquantitative polymerase chain reaction analysis revealed that HPV-16 and -32 DNA sequences are in the episomal state. CONCLUSIONS: This is the first report of BP associated with HPV-32 (the causative agent of a benign hyperplasia) and a rare case of BP associated with 2 HPV types in the same patient. The immune depression could have modified the normal benign progression of the extragenital lesions.     

DNA analysis indicates patient-specific human papillomavirus type 16 strains in Bowen's disease on fingers and in archival samples from genital dysplasia

Human papillomavirus (HPV) type 16 is casually involved in the pathogenesis of anogenital cancer and has also been demonstrated in some patients with Bowen's disease (BD) on the fingers. From two women with HPV 16 in BD on the fingers, and in archival samples from genital dysplasia, collected as long as 26 years ago, the non-coding region of the virus was amplified by the polymerase chain reaction and sequenced. The HPV 16 DNA sequences found in the finger lesions and in the genital archival samples showed no diversities within single patients. Compared with an HPV 16R reference sequence, one patient showed a unique T nucleotide at position 78, whereas the other patient exhibited T and A nucleotides at positions 7193 and 7521, respectively. In one of the patients, the same strain of HPV 16 was found in a digital tumour 26 years after its clearance from the genital tract. DNA sequence analysis indicated patient-specific HPV 16 strains. Auto-inoculation from the genital tract was favoured as a plausible explanation of why HPV 16 caused BD on the fingers.     

Nestin expression in Bowen's disease and Bowen's carcinoma associated with human papillomavirus

Human papillomaviruses (HPVs) have been detected in lesions of Bowen's disease (BD) and Bowen's carcinoma (BC); the invasive tumor retains the cytological characteristics of BD. Previous reports suggest that nestin-expressing hair follicle stem cells are undifferentiated and pluripotent, and nestin expression in some tumors indicates poor differentiation and high grade of malignancy. We identified HPV-DNA in BD (n=25) and BC (n=23) by in situ hybridization (ISH) analysis with INFORM(?) HPV III (Ventana Medical Systems. AZ, USA) and determined nestin expression by indirect immunohistochemical staining with anti-nestin polyclonal antibody (IBL, Gunma, Japan). We detected HPV-DNA in 68% of BD and in 87% of BC. In BD, 13 cases demonstrated the punctuate pattern, and four showed nestin expression. In BC, 19 cases showed the punctuate pattern and 16 showed nestin expression. HPV-DNA integrates into the host genome, and this is observed as the punctuate pattern on ISH. The nestin expression was statistically high in group of BC than BD (P<0.01). These results therefore suggest that HPV-DNA integrated in the genome of tumor cells of these diseases and contributed to malignant alteration. From the standpoint of tumorigenesis, BC might represent one type of poorly differentiated, high-grade squamous cell carcinoma.     

Expression of minichromosome maintenance 5 protein in proliferative and malignant skin diseases

BACKGROUND: The entire minichromosome maintenance (MCM) family (MCM2-7) play roles in the initiation and elongation of DNA replication. Many studies have demonstrated that MCM proteins may be better indicators of a wide variety of proliferative or cancer cells in malignant tissues. OBJECTIVES: To characterize the pattern and frequency of MCM5 expression in proliferative and malignant skin diseases in comparison with those of proliferating cell nuclear antigen (PCNA). METHODS: Twelve normal skin specimens, 12 specimens of psoriasis, 21 specimens of bowenoid papulosis (BP), 16 specimens of Bowen's disease (BD), 38 specimens of skin squamous cell carcinoma (SCC), and 11 specimens of basal cell carcinoma (BCC) were subjected to immunohistochemical staining for MCM5 and PCNA. Results MCM5 protein was expressed in the lower layers of epidermis in psoriasis, while MCM5 protein were present throughout the tumor cells in BP, BD, and moderately/poorly differentiated SCC. MCM5 protein was preferentially expressed in the periphery of well-differentiated SCC or bigger nests of BCC, although some small nests of BCC seemingly showed diffuse staining patterns. The percentages of MCM5-positive cells were 15.7% in normal skin, 21.8% in psoriasis, 75.9% in BP, 83.8% in BD, 63.5% in well-differentiated SCC, 77.5% in moderately differentiated SCC, 79.8% in poorly differentiated SCC, and 21.2% in BCC in average. Well-differentiated SCC showed a significantly lower percentage of positive cells than did moderately differentiated SCC or poorly differentiated SCC. MCM5 staining basically show a similar staining pattern to that of PCNA, but more cells tended to be stained with MCM5 than with PCNA. CONCLUSIONS: Our results demonstrate pattern and frequency of MCM5 expression in various skin diseases and suggest that MCM5 may be a useful marker to detect cell proliferation in skin tissue sections.