尖锐湿疣凹空细胞的观察

本文对９７例尖锐湿疣（ＣＡ）进行聚合酶链反应检测人乳头瘤病毒（ＨＰＶ）ＤＮＡ及光镜下观察凹空细胞的特征。结果显示，９５例有凹空细胞，占９７．９４％。凹空细胞多分布在棘层的中、上层。根据核的改变将凹空细胞分为九型，以镰刀形核、毛虫样核和核大浓染型出现例数最多，分别为７５／９５、７４／９５和６１／９５，此三型对ＣＡ最有诊断价值。ＨＰＶ６、１１阳性９３例（９５．８８％），ＨＰＶ１６、１８、３１、３３、５２ｂ、５８阳性２例（２．０６％），ＨＰＶ６、１１、１６、１８、３１、３３、５２ｂ、５８阴性２例（２．０６％），但有典型的凹空细胞。２例ＣＡ无凹空细胞而ＨＰＶ６、１１为阳性。     

尖锐湿疣人乳头瘤病毒颗粒、核酸及抗原检测

42例尖锐湿疣进行了HPV-DNA序列,HPV抗原和电镜检查。36例(85.7%)HPV-DNA序列检测阳性。其中单一型感染9例,混合型感染27例。以HPV_(11)阳性最多,计32例,HPV_(6b)26例,HPV_(16)19例,HPV_(18)6例。HPV抗原阳性28例(66.7%)。15例查见病毒颗拉(35.7%)。总计阳性率为90.5%(38/42例)。文中对三项检查结果作了比较和分析。     

尖锐湿疣HPV导流杂交快速基因分型及临床意义

目的：探讨外生殖器及子宫颈尖锐湿疣患者皮损脱落细胞或组织中人乳头瘤病毒(Human PapillomaVirus，HPV)9种常见的基因亚型感染分布状况及临床意义。方法：采集患者生殖器或子宫颈尖锐湿疣组织或脱落细胞样本，分离DNA，PCR扩增后与9种常见HPV亚型探针进行导流快速反斑点印迹杂交检测HPVDNA。结果：经PCR扩增并通过通用探针杂交分析后，108例患者标本中，101例标本呈HPVDNA阳性，7例HPV阴性。HPVPCR阳性标本中，98例成功分型。5种高危型HPV16，18，3l，33，58的检出率分别为31．5％(34／108)、16．7％(18／108)、6．5％(7／108)，4．6％(5／108)，13．0％(14／108)。三种低危型HPV6，11，53型的检出率分别为41．7％(45／108)，56．5％(61／108)和6．5％(7／108)。危险度待定HPV亚型CP8304检出率为4．6％(5／108)。结论：结论HPV11、6、16型感染是尖锐湿疣的主要型别。     

A comparison of histopathologic diagnosis and the demonstration of human papillomavirus-specific DNA and proteins in penile warts.

The extent to which the clinical diagnosis of male condylomata acuminata (CA) will be improved by histopathologic examination, immunohistochemical demonstration of papillomavirus common antigen (BPV), or demonstration of human papillomavirus (HPV)-specific DNA was studied. The relation of nuclear atypia to local cytodestructive therapy and specific HPV types was also analyzed. Altogether, the diagnosis could be histologically verified in 116 of 133 (87%) patients with clinically suspected penile CA. Of these, 69 (59%) had a positive result in in situ DNA hybridization for one or more of HPV types 6/11, 16/18, 31/33/35, or 31/35/51. Atypical cells were observed in 53 CA biopsies (46%), and there was a statistically significant correlation between this finding and the high risk HPV types 16/18, 31/33/35, and 31/35/51. Seventeen patients had neither histologic signs of condyloma acuminatum, nor detectable BPV antigen, and in situ hybridization showed HPV type 31/33/35 DNA in one biopsy, and HPV type 6/11 DNA in another. No correlation between the atypical changes and the type of previous therapy for the warts was found. Our results indicate that histopathologic examination supplemented with new HPV-specific methods is an important tool in diagnosing HPV infections.     

尖锐湿疣皮损中人乳头瘤病毒基因分型研究

目的 采用反向杂交研究尖锐湿疣皮损中人乳头瘤病毒(HPV)的感染状况。方法 提取尖锐湿疣新鲜标本的HPVDNA,采用PGMY09/11引物系统进行聚合酶链反应(PCR)。PCR产物在标记有37种HPV型特异性探针的尼龙膜条带上进行HPVDNA杂交分型。所有DNA模板采用HPV6和11型特异性引物进行PCR检测验证。数据经SPSS11.0统计软件分析。结果 杂交结果显示201例标本HPVDNA均为阳性,共发现31种HPV基因型,其主要的HPV基因型名称及所占比例分别如下:HPV11(53.7%,108/201)、HPV6(43.8%,88/201)、HPV16(6.5%,13/201)、HPV52(6.0%,12/201)、HPV33和HPVcp6108(均为5.5%,11/201)、HPV42(5.0%,10/201)等。60.2%(121/201)的标本由单一型HPV感染,39.8%的标本由混合型HPV感染。HPV6和11型特异性引物PCR结果显示HPV6和11的阳性率分别为45.8%和56.2%,与杂交结果比较,一致性分别为98.5%和96.5%,资值分别为0.97和0.93,P值均<0.001。结论 至少有31种HPV基因型与尖锐湿疣相关。HPV11阳性率最高,HPV68、40、54、67、73、82、35、64和83在尖锐湿疣中少见,HPVcp6108在尖锐湿疣中首次发现,且阳性率较高(与HPV33并列第5位)。HPV26、69、70、71、72和IS39可能与尖锐湿疣不相关。尖锐湿疣中单一型和混合型HPV阳性率分别为60.2%(121/201)和39.8%(80/201)。     

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。     

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.     

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。     

Extragenital bowenoid papulosis associated with atypical human papillomavirus genotypes

BACKGROUND: Bowenoid papulosis typically appears as grouped violaceous or red-brown papules in the genital or perianal regions and clinically resembles condylomata acuminata. Isolated extragenital bowenoid papulosis is rare and has been reported in only a few case reports. OBJECTIVES: A 51-year-old immunocompetent, healthy woman had two solitary papules on the elbow; a 41-year-old HIV-positive man had a solitary cutaneous plaque on the abdomen. No genital, periungual, or other extragenital sites of involvement were noted in either patient. The diagnosis was confirmed histologically in both cases. Lesional skin from the female patient was tested with the Digenehybrid HPV DNA assay and was positive for a mixture of low-risk HPV subtypes (6, 11, 42, 43, 44). Lesional skin from the male patient was tested with polymerase chain reaction (PCR). Consensus primers targeted for the HPV L1 region, which is a highly conserved sequence common to more than 20 HPV subtypes encoding a viral capsid protein, were used. PCR using the consensus primers was positive, but type-specific probes for HPV types 6, 11, 16, 18, 45, 31, 33, 35, and 39 were negative. CONCLUSIONS: To our knowledge, our male patient represents the first case of isolated bowenoid papulosis of the abdominal skin. Isolated upper-extremity bowenoid papulosis in our female patient is also a unique case in both location and involvement of low-risk HPV types (6, 11, 42, 43, 44), which have not been previously associated with extragenital bowenoid papulosis.     

Detection of human papillomavirus in verrucous carcinoma from HIV-seropositive patients

Anogenital squamous cell carcinoma has been noted with increased frequency in HIV-seropositive patients. Verrucous carcinoma is a variant of squamous cell carcinoma that tends to be locally invasive and non-metastasizing. Although human papilloma-virus (HPV) has been strongly implicated in other squamous neoplasms, it has been variably associated with verrucous carcinoma and has not been examined in these lesions in the HIV-positive population. The aim of this study was to examine the association of HPV with anal verrucous carcinoma in patients with the human immunodeficiency virus (HIV). HPV DNA in situ hybridization for HPV Types 6/11, 16/18, and 31/33/35 was performed on formalin-fixed, paraffin-embedded tissue from six cases of verrucous carcinoma and four cases of condyloma acuminatum in perianal specimens from HIV-seropositive patients. HPV DNA sequences were identified in five of six cases of verrucous carcinoma and in all cases of condyloma acuminatum. Of the five verrucous carcinomas that harbored detectable HPV DNA, four contained HPV 6/11 and two contained HPV 16/18. One contained both HPV 6/11 and HPV 16/18. All four cases of condyloma acuminatum were positive for HPV 6/11. One patient included in this series had three chronologically separate verrucous carcinomas. The initial lesion was negative for HPV DNA. Subsequent verrucous carcinomas were positive for HPV type 6/11 and type 16/18, respectively. The data presented support the concept that verrucous carcinoma in the HIV-seropositive population is associated with HPV, which may indeed play an important role in its pathogenesis.     

尖锐湿疣患者疣体HPV多型别原位杂交检测

目的　探讨原位分子杂交技术多型别检测尖锐湿疣患者疣体HPV DNA的敏感性。方法　应用原位杂交法 ,通过广谱生物素标记的HPV DNA探针 ,对尖锐湿疣患者疣体HPV 6,11,16,18,3 0 ,3 1,3 3 ,3 5 ,45 ,5 1,5 2型DNA进行检测。结果　在 3 5例标本中 ,有 3 3例HPV DNA呈阳性反应 ,阳性率 94.3 %。阳性反应物主要分布于表皮浅层空泡细胞的细胞核内 ,多呈片状或灶状分布。结论　用广谱生物素标记的原位杂交技术 ,可检测HPV多型别 ,具有敏感性高、特异强的优点 ,并能进行病毒的组织学定位     

Papillomavirus infection among abortion applicants and patients at a sexually transmitted disease clinic.

The base-line prevalence of human papillomavirus (HPV) infection among applicants for first trimester induced abortion and among a sexually transmitted disease (STD) clinic population with macroscopically visible condyloma is investigated. Cervical cells were collected from 505 women applying for induced abortion. Cell scrapes were obtained from the surface of the warts from 81 female and 32 male STD patients. HPV DNA 6/11, 16/18, and 31/33/35 were detected by a dot blot technique (ViraPap and ViraType, Life Technologies, Gaithersburg, MD). Of the 505 abortion applicants, 31 (6.1%) had HPV DNA. In the STD population, 64 (79.0%) patients had positive test results for HPV DNA from cervical, introital cell scrapes, or both. Of the 32 male STD patients from whom preputial, urethral samples, or both were taken, 24 (75%) had HPV DNA. After typing, the relative proportion of HPV 6/11 was 9.7% in abortion applicants, 72.2% in female STD patients, and 91.3% in male STD patients. The relative proportion of high-risk HPV types, such as 16/18 and 31/33/35, was 89.7% among abortion applicants, 38.9% among female STD patients and 13.0% among male STD patients. The majority of abortion applicants infected with HPV were thus infected with a potentially oncogenic HPV type, and the prevalence of HPV type 16/18 and 31/33/35 was also high among female STD patients.     

Simultaneous human papillomavirus 6 (HPV 6) -positive condyloma acuminatum, HPV 31-positive Bowen's disease, and non HPV-associated extramammary Paget's disease coexisting within an area presenting clinically as condyloma acuminatum

An 83-year-old Japanese man presented with multiple verrucous papules clustering on a plaque located on the frontal aspect of the scrotum. Histologically, there were three distinct epithelial changes compatible with condyloma acuminatum, Bowen's disease, and extramammary Paget's disease (EMPD). By in situ hybridization, the zone of condyloma acuminatum was positive for HPV 6 and well demarcated from HPV 31-positive Bowen's disease. EMPD was negative for targeted HPV 6/11/16/18/31/33 probes. Immunohistochemically, Paget's cells expressing cytokeratin 7 were distributed as scattered single cells or clusters mainly in the lower part of the HPV 6/31-positive epithelium. To the best of our knowledge, this is the first reported case of the occurrence of condyloma acuminatum, Bowen's disease, and EMPD within the same lesion.     

Human papillomavirus-associated induction of human β-defensins in anal intraepithelial neoplasia

Background  Antimicrobial peptides and proteins (AMPs) are widely distributed effector molecules of the innate immune system with well-known antibacterial activity. However, there is a paucity of information regarding antiviral effects of AMPs.
Objectives  The present study was performed to analyse expression of AMPs in human papillomavirus (HPV)-associated anal skin lesions of human immunodeficiency virus (HIV)-positive men who have sex with men (MSM), a special high-risk group for persistent HPV infections and anal dysplasia.
Methods  Skin lesions were analysed for the presence of LL-37, RNase 7, and human β-defensin (hBD)-1, hBD-2 and hBD-3. Moreover, HPV typing and HPV DNA load determination for HPV types 6, 11, 16, 18, 31 and 33 were performed to evaluate possible correlations between expression of AMPs and lesional HPV types.
Results  Skin biopsies of 45 HIV-positive MSM with anal intraepithelial neoplasia (AIN), anal condylomata acuminata or unaffected anal mucosa, as well as condylomata acuminata of eight HIV-negative MSM, were analysed for AMP mRNA expression. Additionally, immunohistochemical analysis for hBD-2 and hBD-3 was performed in a total of 45 samples. hBD-2 and hBD-3 gene and protein expression was significantly increased in both AIN and condyloma, whereas LL-37, RNase 7 and hBD-1 gene expression did not differ significantly from unaffected anal mucosa. AMP expression correlated neither with the number of HPV types nor with the high-risk and low-risk HPV DNA loads of the quantified types. No significant differences in AMP expression were observed in condylomata of HIV-positive and HIV-negative MSM.
Conclusions  hBD-2 and hBD-3 expression was shown to be significantly upregulated in HPV-associated anal skin lesions of both HIV-positive and HIV-negative MSM. Their biological significance in the innate immunity against these lesions needs further research.     

Papillomavirus-induced anogenital lesions in 121 HIV seropositive men. Clinical, histological, viral study, and evolution]

OBJECTIVE: To determine the prevalence of the Human Papillomavirus (HPV) in Human Immunodeficiency Virus (HIV) infected men, using clinical examination and molecular hybridization in situ. PATIENTS AND METHODS: From May 1995 to May 1997 we studied the prevalence, clinical and histological characteristics, the types and the evolution of the HPV lesions among 121 HIV-infected men. The HPV DNA was determined by molecular hybridization in situ, using biotinylated probes which recognized HPV types 6/11, 16/18 and 31/33/35 in 79 p. 100 (5/19) of the patients (17 biopsies). RESULTS: Sixteen per cent (19/121) of the patients are HPV infected: genital warts in 37 p. 100 (7/19), anal warts in 37 p. 100 (7/19), and ano-genital warts in 26 p. 100 (5/19) of the patients. In every case of anal codyloma, intracanalar lesions were found. In 47 p. 100 (9/19) of the cases, histological exam showed an intra-epithelial neoplasia. The HPV types 6/11, 16/18 and 31/33/51 were positive in 53 p. 100 (9/17), 35 p. 100 (6/17) and 35 p. 100 (6/17) biopsies respectively. High-risk types of HPV have been noted in 71 p. 100 (12/17) of the biopsies. The evolution of the clinical lesions was: recovering in 47 p. 100 (9/19) of the patients (after 3 months of treatment), recurrence in 16 p. 100 (3/19) of the anal warts (after 1 to 3 months of treatment), stabilization in 16 p. 100 (3/19) of the genital warts (after 6 months of treatment) and extension in 11 p. 100 (2/19) of the anogenital warts (after 3 months of treatment). CONCLUSION: The high prevalence of condyloma and dysplasia emphasizes the importance of the anogenital exam in HIV-positive patients. In case of anal lesions, anuscopy and biopsy are required. We insist on the need to closely follow these patients with HPV lesions in order to adapt treatment. Anal cytology and HPV-DNA detection by Hybrid Capture Assay, should be developed for screening and prevention of the malignant transformation of HPV lesions in this population.     

Common Association of HPV 2 with Anogenital Warts in Prepubertal Children

Abstract: Anogenital (AG) warts in 31 prepubertal children were HPV typed by nonisotopic in situ hybridization (NISH) using digoxigenin labeled probes for human papilloma virus (HPV) types 1–5, 6,11,16,18, 31, and 33. Mode of transmission was determined from historical, clinical, and laboratory data independent of HPV typing. HPV 2 was detected most commonly (13/31 warts) followed by HPV 6 (7/31), HPV 11 (5/31), and HPV 16 (1/31). Although not reaching statistical significance, our results suggested that a mucosal HPV type (6,11,16) in a child's AG warts implied transmission from mucosal warts and conversely cutaneous HPV 2 transmission from warts at a cutaneous site. HPV typing provided no helpful information regarding actual mode of transmission of AG warts In these children. The high prevalence of HPV 2 in children's AG warts and the low prevalence of sexual abuse (2 of 31 children) found in this study suggest innocent auto- or heteroinoculation from cutaneous warts may be a common means by which children acquire AG warts.     

临床表现为假性湿疣的高危型HPV感染

目的 探讨12例临床表现酷似假性湿疣的HPV感染的临床及HPV分型特点.方法 检测12例有非婚性接触史临床表现为外阴假性湿疣的女患者,以反向杂交法检测活检或局部刮除物中HPV核酸扩增并分型.结果 12例HPV全部阳性.12例患者中共查出13种HPV亚型.其中3种低危型11、42、43,10种高危型16、18、31、33、35、39、51、52、58、59.每例患者至少有1种以上高危HPV亚型.4例患者为1种HPV亚型的感染,6例患者为2种HPV亚型的感染,另有两例分别有3或4种HPV亚型的感染.结论 在临床表现为假性湿疣患者中有可能包含高危型HPV的感染.     

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。     

温州地区尖锐湿疣组织人乳头瘤病毒检测及型别分析

用人乳头瘤病毒（ＨＰＶ）通用引物（ＧＰ）及型特异性引物（ＳＰ）６、１１、１６、１８对４２例尖锐湿疣（ＣＡ）组织和４８例对照组进行多聚酶链反应（ＰＣＲ）检测。结果：ＣＡ组织五组引物扩增阳性率分别为８５．７％（３６／４２）、７３．８％（３１／４２）、１９．１％（８／４２）、２．４％（１／４２）、４．８％（２／４２），多重型别阳性率为１１．９％（５／４２）；对照组ＨＰＶ阳性率为６．３％（３／４８）。表明本地区ＣＡ以ＨＰＶ６型感染为主     

尖锐湿疣组织HPV基因芯片分型及型别分析

目的研究尖锐湿疣(CA)组织中人乳头瘤病毒(HPV)感染的病毒亚型分布,CA患者中高危型HPV感染的情况。方法采用基因芯片检测和分型方法,对武汉市302例CA患者的皮损组织进行HPV检测及分型。结果302份CA组织标本中HPV阳性286例,检出率为94.7%,单一型别阳性率为73.8%,多重型别阳性率为20.7%,低危型别(HPV6,11,42,43,44)阳性率为80.5%,高危型别(HPV16,18,31,33,35,39,45,51,52,53,56,58,59,66,68,73,83,MM4)阳性率为14.2%,对照组阳性率为16.7%。结论本地区尖锐湿疣以单一型别及低危型别HPV感染为主,高危型HPV感染阳性率为14.2%。