6例分化型外阴上皮内瘤变临床病理观察

目的：探讨分化型外阴上皮内瘤变( differentiated-type vulvar intraepithelial neoplasia, dVIN)的临床病理学特征、鉴别诊断、治疗及预后。方法回顾性分析6例dVIN的临床特点、病理形态学和免疫表型特征,并复习相关文献。结果患者均为女性,年龄53~80岁,平均62岁,临床特点大多表现为外阴白斑、激惹、瘙痒、疼痛、溃疡、触血。病理组织学特征表现：基底与副基底层显著增生,上皮角下延、吻合,细胞显著异型,核仁明显,核分裂活跃,伴异常角化;中、表层细胞高度分化,细胞间桥显著,伴明显的胞质嗜酸性改变,表层角化过度与角化不全;邻近上皮真皮水肿与胶原化,带状淋巴细胞浸润,表皮增生与角化过度。免疫表型：p53在dVIN中的阳性率为83．3%(5/6),p16不表达(0/6),dVIN基底层和副基底层细胞Ki-67增殖指数>90%。4例随访时间为6~36个月,平均17个月,其中1例术后9个月死亡,1例术后6个月复发,复发和死亡的2例同时或异时伴浸润性鳞状细胞癌,其余2例术后分别随访18个月和36个月,均无复发。结论 dVIN是一种少见的高级别外阴上皮内病变,具有高的、潜在进展危险性,p53、p16及Ki-67联合使用有助于dVIN的诊断。     

p16、p53和Ki-67蛋白在宫颈上皮内瘤变中的表达及意义

目的 探讨p16、p53和Ki-67蛋白在宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)中的表达及临床意义.方法 采用免疫组化SP法检测正常子宫颈或炎性病变组织、CIN1～3中p16、p53和Ki-67蛋白的表达.结果 p16、p53和Ki-67蛋白在正常子宫颈或炎性病变中罕见表达,在CIN1～3组织中三者表达均较高,随CIN级别升高p16、p53和Ki-67表达增强,各组间表达差异有统计学意义(P<0.05).同时p16、p53和Ki-67三者阳性表达均可见分层现象,在CIN1中大部分阳性细胞位于子宫颈鳞状上皮的下1/3,在CIN2中多累及上皮下2/3,而CIN3则普遍超过上皮的下2/3或全层弥漫阳性,各组间差异具有统计学意义(P<0.05).结论 p16、p53和Ki-67蛋白表达均与子宫颈上皮内瘤变的病变进展密切相关,联合检测p16和Ki-67的抗原表达可作为CIN分级诊断的辅助方法,具有较好的应用价值.     

肺鳞癌人乳头状瘤病毒感染的原位杂交检测和观察

经多重多聚酶链反应，检测４９例肺鳞癌中发现７例人乳头瘤病毒阳性的肿瘤组织，采用生物素标记ＨＰＶＤＮＡ探针，进行原位杂交检测，结果发现在５例肿瘤组织中显示ＨＰＶＤＮＡ阳性信号，其中ＨＰＶ１１型阳性３例；ＨＰＶ１６例阳性１例；１例为ＨＰＶ１１例和１６型均阳性。原杂交ＨＰＶＤＮＡ阳性信号，大多位于凹空样肿瘤细胞或低分化鳞癌细胞的核内，分子生物学研究表明ＨＰＶ感染可能与部分鳞有关。     

女性下生殖道癌临床病理特征与人乳头状瘤病毒型别间的相关性研究

目的：探讨女性下生殖道癌的临床病理与人乳头状瘤病毒（HPV）型别之间的关系。方法：回顾性研究100例下生殖道癌（宫颈癌63例,外阴癌37例）的临床病理特征,并应用PCR技术检测每份标本的HPV状态。结果：在模板内参照阳性的87份中,宫颈癌54份,外阴癌33份,HPV阳性率在宫颈癌中为88．3％,以HPV16型（55．6％）和HPV18型（24．4％）为主,在33例外阴鳞癌中,HPV阳性仅见于基底细胞癌和混疣样癌,阳性率均为83．3％,以HPV16型为主（70．0％）;6例基底细胞样癌中有3例合并宫颈鳞状上皮肿瘤,其中2例宫颈与外阴的肿瘤均为HPV16型阳性;21例角化鳞癌则未检测出HPV－DNA,但有发病年龄高（平均63．3岁）。形态学上角化明显和预后较差等特征。结论：HPV16型和18型在宫颈癌中性阳率较高,而在外阴癌中HPV阳性的意义则因组织学类型而不相同。     

人乳头状瘤病毒感染及感染拷贝数与食管癌的关系

人乳头状瘤病毒感染及感染拷贝数与食管癌的关系吕丽春沈忠英游绍进沈健蔡唯佳一、材料和方法收集１９９４年～１９９５年我院附属医院外科手术切除、病理确诊的新鲜食管癌标本５５例，每例分别取癌组织、癌旁粘膜和远端切缘粘膜３部分，经ＯＣＴ包埋在一起，常规冰冻切片...     

人乳头状瘤病毒感染及其亚型与鼻内翻性乳头状瘤相关性探讨

目的 了解不同亚型的人乳头状瘤病毒(human papillomavirus,HPV)在鼻内翻性乳头状瘤(nasal inverted papilloma,NIP)中的分布,探讨其在NIP发病中的作用.方法 将78例NIP腊包埋组织标本分为单纯NIP组、伴不典型增生NIP组和NIP癌变组,同时选取40例健康体检者鼻腔粘膜组织作为健康对照组,采用聚合酶链反应(PCR)技术检测HPV6、11、16及18型感染情况.结果 78例NIP石蜡包埋组织标本HPV总阳性率为61.5％.单纯NIP组、伴不典型增生NIP组和NIP癌变组阳性率分别达59.5％、73.9％和46.2％.随着NIP临床分期级别的升高,HPV感染率也呈相应的上升趋势.单纯NIP组以低危型HPV6、11型感染为主,NIP癌变组则同时检出高危型HPV16型和HPV18型.结论 HPV感染在NIP发病中起着重要作用,其中高危型HPV感染能扩大NIP病变范围,而HPV16、18型可能与NIP恶变有关联.     

宫颈上皮内瘤变及其治疗对妊娠的影响

宫颈鳞状上皮内瘤变是一种和HPV感染有关的宫颈癌前病变。根据病变程度分为低度和高度鳞状上皮内病变,其中低度病变包括HPV感染和C IN1,高度病变包括C IN2~3。HPV感染和宫颈上皮内瘤变是一种生育年龄妇女常见的妇科疾病,HPV感染、宫颈病变本身及其治疗对妊娠的影响倍受关注。HPV感染在妊娠期间可能增加,经阴道分娩者新生儿暴露于HPV的机会增多,尚不能下结论对所有HPV感染者均采用剖宫产的分娩方式。妊娠期间发现C IN1,可以观察并产后随访。如果为C IN2~3,妊娠可能不会加重病变程度,但对阴道镜检查不满意者或高度怀疑浸润癌者应在孕期明确诊断,可于妊娠中期行宫颈锥切术。妊娠中期行宫颈锥切术可能使剖宫产率增加。C IN保守治疗对于患者受孕能力无显著的影响;宫颈冷刀锥切及LEEP锥切可能增加早产率,早产率的增加可能和胎膜早破的发生有关。     

134例宫颈上皮内瘤变患者HPV分型检测结果分析

目的探讨宫颈上皮内瘤变(CIN)中不同类型人乳头瘤病毒(HPV)的感染及分型情况。方法回顾性研究北京军区总医院妇科门诊134例已经确诊的CIN患者,采用凯普的核酸分子快速导流杂交基因芯片技术对HPV进行分型。结果 1.各级别CIN以HPV高危亚型单一或者多重感染为主;2.在HPV阳性的101例患者中,单一感染者73例(72.28%),双重感染者24例(23.76%),三重感染者3例(2.97%),四重感染者1例(0.99%);3.HPV-16、58感染率最高,分别为30.83%,24.06%。结论 HPV感染以高危型和多重感染为主;随CIN级别的增高,HPV阳性率也是增高的;HPV-16亚型的感染率最高。     

应用基因芯片诊断宫颈石蜡组织样本中人乳头状瘤病毒感染及其临床意义

目的探讨应用基因芯片检测宫颈石蜡组织标本中人乳头状瘤病毒（HPV）感染的可能性及其临床意义。方法收集解放军总医院诊断为宫颈鳞状上皮病变的石蜡组织标本40例,其中宫颈浸润性鳞癌18例,宫颈上皮内瘤变（CIN）Ⅲ12例,CINⅠ4例,CINⅡ6例。从组织中提取DNA后采用基因芯片检测23种常见HPV基因亚型,即PCR扩增后产物在基因芯片上进行杂交。同时选用10例经基因芯片检测16型和18型基因阳性的宫颈鳞癌的石蜡组织切片做原位杂交。基因芯片检测结果与部分原位杂交结果进行比较并分析。结果基因芯片检测的18例宫颈鳞癌HPV高危亚型均为阳性（100％）,其中1例为混合阳性;12例CINⅢ中11例为高危亚型阳性（91．7％）,1例阴性;6例CINⅡ的宫颈病变中高危型5例阳性,低危型1例阳性;4例CINⅠ中有2例低危型阳性、2例阴性;宫颈鳞癌和CINⅢ组与CINⅠ和Ⅱ组比较,差异有统计学意义（U=80．0,P〈0．01）。10例宫颈鳞癌基因芯片HPV16型和18型阳性组织中,原位杂交同型探针6例检测显示阳性。结论HPV基因芯片技术可用于检测多种亚型,特异性强,敏感性高,对HPV感染亚型的鉴别及宫颈癌的预防和治疗具有重要意义。     

p16INK4a在宫颈细胞学鳞状上皮内瘤变中的意义

目的探讨p16^INK4a的表达在宫颈细胞学中鳞状上皮内瘤变中的意义及其与人乳头状瘤病毒（HPV）型别之间的关系。方法对88例经活检证实的液基细胞学标本[包括20例慢性宫颈炎、18例低度鳞状上皮内瘤变（LSIL）、34例高度鳞状上皮内瘤变（HSIL）及16例鳞状上皮细胞癌（SCC）],分别用免疫细胞化学（EnVision方法）检测其p16^INK4a的表达,并对所有标本用聚合酶链反应（PCR）方法检测HPV型别（包括HPV16、18、31、33、35、39、45、51、52、53、56、58、59、66、68、6、11、42、43及44型）。结果p16^INK4a在慢性宫颈炎组呈阴性,在LSIL、HSIL及SCC组的表达率分别为27．8％、100％及100％,LSIL、HSIL及SCC组p16^INK4a的表达均显著高于慢性宫颈炎组（P〈0．01）;p16^INK4a在高危型HPV感染组的表达率（96．4％）显著高于低危型HPV感染组（7．7％）,差异有统计学意义（t=4．32,P〈0．01）。结论p16^INK4a是一种敏感性较高,特异性较好的标记物,可以识别与高危型HPV有关的非典型鳞状上皮细胞。     

Specific intraepithelial localization of mast cells in differentiated vulvar intraepithelial neoplasia and its possible contribution to vulvar squamous cell carcinoma development

van de Nieuwenhof H P, Hebeda K M, Bulten J, Otte‐Holler I, Massuger L F A G, de Hullu J A & van Kempen L C L T
(2010) Histopathology 57, 351–362
Specific intraepithelial localization of mast cells in differentiated vulvar intraepithelial neoplasia and its possible contribution to vulvar squamous cell carcinoma development Aims: The aetiology of vulvar squamous cell carcinomas (SCC) that are not causally associated with high‐risk human papillomavirus remains largely elusive. The aim of this study was to analyse the inflammatory response in its presumed precursor lesions, lichen sclerosus (LS) and differentiated vulvar intraepithelial neoplasia (dVIN), and provide evidence that dVIN is a likely precursor of vulvar SCC. Methods and results: Immunohistochemical analyses for CD4+, CD8+, CD20+, CD68+, S100+ and tryptase‐positive immune cells were performed and quantified in LS (n = 7), dVIN (n = 19), SCC (n = 11), and normal vulvar tissue (n = 8). The subepithelial inflammatory response in dVIN and SCC was comparable, but absent in LS. Abundant intraepithelial mast cells were observed in dVIN only, and confirmed by electron microscopy, toluidine blue staining and cKIT expression. Adjacent keratinocytes displayed increased proliferation as determined by MIB‐1 positivity. Electron microscopy revealed intraepithelial mast cell degranulation. Intraepithelial mast cells were not or infrequently observed in vulvar hyperplasia (n = 13), condylomata acuminata (n = 5), keratinocytic intraepidermal neoplasia of sun‐exposed skin (n = 15), epidermal hyperplasia of head and neck (n = 12), and psoriasis (n = 3). Conclusions: These data indicate that dVIN can be recognized by intraepithelial mast cells and that they might promote the progression of dVIN to SCC.     

细胞周期素D1和抗原Ki-67在宫颈上皮内瘤样病变和宫颈鳞癌的表达及与人乳头瘤病毒感染转归的关系

目的 研究细胞周期素D1(cyclin D1)、抗原Ki-67在宫颈上皮内瘤样病变(CIN)和宫颈鳞癌发生发展中作用及其与人乳头瘤病毒(HPV)感染转归的关系.方法 2002年1月至2006年12月广州医学院第一附属医院HPV阳性患者104例,分2组:(1)研究组:82例,即病理确诊CIN Ⅰ组17例、CINⅡ组19例、CINⅢ组23例、宫颈鳞癌组23例.(2)对照组:柱状上皮异位22例.应用EnVision法检测宫颈病变组织中cyclin D1、Ki-67蛋白的表达,杂交捕获试验检测宫颈分泌物或阴道残端中HPV感染情况,随访各组患者术后1年内的HPV变化.结果 (1)cyclin D1在各组宫颈组织细胞核内均有表达.其阳性率CINⅢ组[82.61%(19/23)]、宫颈鳞癌组[86.96%(20/23)]与对照组[27.27%(6/22)]、CINⅠ组[58.82%(10/17)]比较,差异有统计学意义(P<0.05),在宫颈鳞癌组与CINⅡ组[68.42%(13/19)]阳性率比较差异有统计学意义(P<0.05).(2)Ki-67在各组宫颈组织细胞核内均有表达,其对照组阳性率[31.82%(7/22)]与CINⅢ组[86.96%(20/23)]、宫颈鳞癌组[91.30%(21/23)]比较差异有统计学意义(P<0.05),而在宫颈鳞癌组与CINⅠ组[58.82%(10/17)]、CIN Ⅱ组[63.16%(12/19)]比较差异有统计学意义(P<0.05).(3)术后1年内各组HPV的转阴率分别与cyclin D1、Ki-67的表达强度呈负相关(rs=-0.299,rs=-0.367,P<0.05).结论 cyclinD1和Ki-67在CIN和宫颈鳞癌的细胞增殖活动中起作用,且两者可能与HPV感染转阴率有关.     

Successful treatment of recurrent vulvar intraepithelial neoplasia resistant to interferon and isotretinoin with cidofovir

Vulvar intraepithelial neoplasias are difficult to eradicate completely without extensive surgical intervention. Cidofovir, a deoxycytidine monophosphate analog, may have a therapeutic role in this disease. A 43-year-old woman with a 20-year history of genital warts presented with extensive vulvar intraepithelial neoplasia III, and refused surgical resection. Topical cidofovir 1% in Beeler base completely eradicated the lesion. Successive treatment applications, however, were necessary. Cidofovir is a promising topical antiviral compound for HPV induced vulvar intraepithelial neoplasia.     

Ki-67 and ProExC are useful immunohistochemical markers in esophageal squamous intraepithelial neoplasia

Esophageal squamous intraepithelial neoplasia has been widely recognized as a precursor lesion for esophageal squamous cell carcinoma. Early detection offers the best prognosis for esophageal squamous cell carcinoma. The differentiation of squamous dysplasia from reactive change and the classification of squamous dysplasia into high-grade or low-grade are sometimes subjective and challenging. In this study, we sought to evaluate multiple biomarkers and to develop clinically useful adjunct tools for difficult esophageal squamous intraepithelial neoplasia cases. Immunohistochemical stains using antibodies against Ki-67, ProExC, p16, and p53 were performed on esophageal biopsy or resection specimens from 25 patients including 35 foci of high-grade dysplasia and 25 foci of low-grade dysplasia, and from 10 control cases containing 52 foci of normal/reactive hyperplasia. In situ hybridization tests for human papillomavirus were performed in 11 cases. The immunostains for all 4 markers were scored as negative, intermediate, and strong according to established criteria. Intermediate and strong Ki-67 and ProExC staining showed similar detecting power and exhibited very high sensitivity and specificity for distinguishing normal/reactive hyperplasia from esophageal squamous intraepithelial neoplasia and normal/reactive hyperplasia from low-grade esophageal squamous intraepithelial neoplasia. Strong Ki-67 staining was exclusively seen in high-grade esophageal squamous intraepithelial neoplasia, which provided additional value in distinguishing high-grade from low-grade esophageal squamous intraepithelial neoplasia. Strong ProExC staining was also seen in most high-grade esophageal squamous intraepithelial neoplasia foci (80%). Although the frequencies of intermediate/strong staining patterns of p53 increased with increasing degree of dysplasia, the sensitivity of p53 was much lower than that of Ki-67 and ProExC. p16 did not show consistent immunostain pattern in the normal/reactive hyperplasia and esophageal squamous intraepithelial neoplasia. Two (18%) of 11 tested cases were positive for human papillomavirus infection. This study demonstrates that both Ki-67 and ProExC can be used as an adjunct tool for diagnosing difficult cases of esophageal squamous intraepithelial neoplasia.     

宫颈瘤变程度与其相对应的HPV基因型分布规律的研究

目的研究不同级别宫颈上皮内瘤变（cervical intraepithelial neoplasia,CIN）与人乳头瘤病毒（Human papil-lomavirus,HPV）基因型分布之间的关系。方法收集321例经组织病理学诊断确定为CIN2及以上（CIN2＋）病人的宫颈分泌物标本,其中CIN2 67例,CIN3/ACIS 247例,浸润性宫颈癌7例。采用深圳亚能生物技术有限公司的人乳头瘤病毒（HPV）基因分型检测试剂盒对所选标本进行HPV检测及基因分型。结果在所有321例标本中,HPV阳性300例,阳性率93.5%,两型及两型以上感染138例,占比43.0%。HPV16最常见,检出率为41.1%（132/321）,其次为HPV31,33,52,18和51。HPV16和33的单型感染在CIN3＋中比CIN2更常见,具有统计学差异（P〈0.05）。在包含有HPV16或HPV33的多型感染中,含有HPV16或HPV33的多型感染在CIN3＋中比CIN2更常见,具有统计学意义（P〈0.05）。结论 HPV16和HPV33比其他型别HPV具有更强的潜在致癌性。     

High-risk human papillomavirus infection of the genital tract of women with a previous history or current high-grade vulvar intraepithelial neoplasia

Human papillomavirus (HPV) infection is associated with high-grade vulvar intraepithelial neoplasia (VIN-3). The prevalence of anogenital HPV infection in women with previously treated VIN-3 has not been documented yet. This cross-sectional study compared high-risk HPV DNA detection rates in women with past (n = 30) and current (n = 22) VIN-3 to those without current or past VIN (n = 86). HPV DNA was detected in vulvar and cervical samples with Hybrid Capture 2 (HC-2). Smoking was associated in multivariate analysis with current VIN-3 (odds ratio (OR) 8.3, 95% confidence interval (CI) 2.0-8.2) and any VIN-3 history (OR 6.5, 95% CI 2.5-16.5). High-risk HPV DNA was found on the vulva of 64%, 33%, and 20% of women with current VIN-3, past VIN-3, and without previous or current VIN, respectively. After controlling for age and smoking, high-risk HPV vulvar infection was associated with cervical high-risk HPV infection (OR 8.6, 95% CI 2.8-26.5; P = 0.001). After controlling for age, HPV infection was more often multifocal in women with current VIN-3 compared to women with previous but no current VIN-3 lesion (OR 17.6, 95% CI 1.4-227.2). Multifocal vulvar HPV infection was detected in women with previous or active VIN-3. Longitudinal studies are required to determine if the multifocality of HPV infection on the vulva could explain the high recurrence rate of VIN-3.     

宫颈上皮内瘤变中p16INK4A和Ki-67的表达

摘要：目的探讨p16^INK4A、Ki-67和HPV抗原表达及高危型HPV（HR—HPV）检测在宫颈上皮内瘤变（CIN）病理诊断中的意义。方法选取宫颈活检病理确诊为CIN的组织蜡块101例,重新切片,应用免疫组化两步法检测p16^INK4A、Ki-67和HPV抗原表达,并取正常宫颈组织50例进行对比研究。同时,应用第二代杂交捕获法对其中25例CIN组织样品进行HR—HPV检测。结果p16^INK4A蛋白表达水平在CINI、CINⅡ、CINⅢ级之间差异有非常显著性（P〈0．001）,其表达水平随着CIN级别的增高而增加,呈现良好的线性相关性（P〈0．001）;Ki-67蛋白表达阳性细胞多少与CIN分级之间无显著相关性（P〉0．05）,但其在宫颈鳞状上皮中的位置分布与CIN级别之间却有显著相关性（P〈0．05）;HPV抗原免疫组化染色阳性反应仅呈现于CIN鳞状上皮表层挖空细胞内,其阳性率在不同级别CIN之间的差异无统计学意义（P〉0．05）;HR—HPV在CINI、CINⅡ和CINⅢ级的检出率分别为81．8％、80．0％和100．0％,但其检出率在不同级别CIN之间差异也无显著性（P〉0．05）。结论p16^INK4A和Ki-67染色对CIN的病理诊断和分级具有一定诊断价值,对于CINI级形态结构不典型的病例,HR—HPV的检测结果对病理诊断有辅助意义。     

The utility of Ki-67 expression in the differential diagnosis of prostatic intraepithelial neoplasia and ductal adenocarcinoma

Cribriform and/or papillary prostatic lesions observed on limited tissue, such as needle biopsy, can pose diagnostic dilemmas. One such area of difficulty is the distinction between papillary and/or cribriform prostatic high-grade prostatic intraepithelial neoplasia (HG-PIN) and ductal adenocarcinoma. Over 48 months, we identified 17 cases of ductal adenocarcinoma and 17 cases of HG-PIN from radical retropubic prostatectomy specimens. The HG-PIN lesions were in all cases associated with an acinar prostatic adenocarcinoma component. For each case, we evaluated the proliferative activity, assessed by Ki-67 immunohistochemistry. The majority (82%) of ductal adenocarcinomas were composed of mixed papillary and cribriform patterns, with the remaining demonstrating pure papillary or cribriform patterns. The HG-PIN lesions showed a papillary, cribriform, or mixed papillary/cribriform architecture. The proliferative activity, defined as Ki-67 labeling index, was statistically higher in ductal adenocarcinoma (mean 33%, range 21%-66%) as compared with HG-PIN (mean 6%, range 2%-15%), with no overlap in the Ki-67 indices (P = 0001). A combination of histological features and measurements of cellular proliferation may be helpful to distinguish HG-PIN from ductal adenocarcinoma in limited prostatic tissue samples.     

p53 immunostaining in lichen sclerosus is related to ischaemic stress and is not a marker of differentiated vulvar intraepithelial neoplasia (d-VIN)

AIM: To analyse p53 immunoreactivity in 207 biopsy specimens of lichen sclerosus (LS) and "differentiated vulvar intraepithelial neoplasia" (d-VIN), a postulated precursor lesion for LS-associated vulvar squamous cell carcinoma (SCC), which is characterized by atypical basal keratinocyte proliferations with p53+ basal/suprabasal keratinocyte nuclei. METHODS AND RESULTS: Forty early, 78 classic, 30 hypertrophic vulvar LS, 26 paediatric vulvar and penile LS, 33 vulvar LS-associated SCC and 30 vulvar/penile control specimens were examined for p53 expression and the presence of d-VIN. Nuclear p53 staining was observed in 175/207 LS biopsy specimens. Eighty percent of early and 69% of paediatric LS showed discontinuous/continuous p53 staining in basal keratinocytes. Classic LS showed no p53 staining in 17%, discontinuous basal keratinocyte staining in 20%, continuous basal keratinocyte staining in 58%, basal/suprabasal staining in 5%. Hypertrophic LS revealed basal keratinocyte staining in 32% and basal/suprabasal staining in 61%. p53 staining was associated with sclerosis of blood vessels and dermis, lymphoid infiltrates, vasculitis and hypertrophic LS. d-VIN was seen in 2% of LS alone and in 24% of LS-associated SCC. CONCLUSION: d-VIN in LS is rare, while p53 staining is common and best explained as an ischaemic stress response due to poor oxygenation, vasculitis and inflammation rather than as a marker of a precancerous lesion in LS.