p16/Ki-67双染在高级别宫颈上皮内瘤变诊断中的应用价值

目的:比较细胞学p16/Ki-67双染检测和高危型人乳头瘤病毒(hrHPV)检测对细胞学为无明确意义的非典型鳞状细胞(ASCUS)患者的分流作用与诊断效能.方法:收集细胞学诊断为ASCUS,同时行hrHPV检测,并有宫颈组织病理学检查结果的患者信息.将剩余宫颈上皮脱落细胞标本进行p16/Ki-67免疫细胞化学双染检测,...     

p16~(INK4a)在液基细胞学筛查宫颈病变中高危人群的临床价值

目的:利用液基细胞学标本探讨p16INK4a在宫颈癌前病变及宫颈癌中的表达。方法:选取104例经活检证实的TCT阳性病例剩余标本,包括9例非典型鳞状细胞-意义不明(ASC-US)、54例低度鳞状上皮内病变(LSIL)、4例非典型鳞状细胞-不除外上皮内高度病变(ASC-H)、33例高度鳞状上皮内病变(HSIL)和4例鳞状上皮细胞癌(SCC),制作液基薄片进行p16INK4a免疫细胞化学检测,同时用基因杂交捕获法Ⅱ(HC-Ⅱ)检测高危人乳头瘤病毒(HR-HPV)DNA,探讨病毒负荷量与宫颈病变级别的关系。结果:p16INK4a在ASC-US、LSIL、ASC-H、HSIL和SCC中的表达率分别是22.2%、40.7%、50%、81.8%和100%。p16INK4a在ASC-US和HSIL之间、LSIL和HSIL之间表达有差异(P<0.003);结合组织学分析54例LSIL标本中p16INK4a阳性率,在炎症、CINⅠ、CINⅡ～Ⅲ中的阳性率分别是28.6%,25.0%和91.7%,CINⅡ～Ⅲ中p16INK4a的阳性率明显高于炎症组和CINⅠ组,3组之间的差异有显著统计学意义(P<0.001);HR-HPV DNA的荷载量与宫颈病变级别无相关性。结论:p16INK4a在液基细胞学HSIL中高表达,是细胞学诊断为LSIL病例中筛查癌前病变高危人群有意义的生物标记物。     

人乳头瘤病毒L1壳蛋白在筛查宫颈鳞状上皮内病变中的应用

目的:探讨人乳头瘤病毒(HPV)L1壳蛋白筛查HPV阳性妇女宫颈脱落细胞中宫颈鳞状上皮内病变的应用价值。方法:选取2012年5月至2014年12月就诊于温州市人民医院的妇女212例,收集宫颈脱落细胞并行HPV L1壳蛋白检测、HPV DNA分型、TCT(液基细胞学)及阴道镜下活检,比较HPV阳性妇女的宫颈脱落细胞中HPV L1壳蛋白的表达情况。结果:212例细胞学标本中HPV L1壳蛋白阳性率为33.9%,其中未见上皮内病变/恶性细胞组(NILM)、无明确诊断意义的鳞状上皮细胞病变组(ASCUS)、低度鳞状上皮内病变组(LSIL)、不能排除高度鳞状上皮内病变组(ASC-H)、高度鳞状上皮内病变组(HSIL)中阳性率分别为47.1%、35.1%、54.2%、29.2%、16.1%,各组比较差异有统计学意义(P0.05);两两比较,HSIL组与LSIL组和NILM组比较,差异均有统计学意义(P均0.005);进行数据合并后,LSIL/ASCUS组与ASC-H/HSIL组比较差异有统计学意义(P=0.001)。178例宫颈细胞学异常患者中,宫颈低级别病变和宫颈高级别病变的HPV L1壳蛋白阳性率比较,在ASCUS组(P=0.000)、LSIL组(P=0.004)中均有差异,在ASC-H组(P=0.127)、HSIL组(P=0.515)中均无差异。HPV 16/18感染患者的HPV L1壳缺失同宫颈高级别病变有更紧密的关系(P=0.003)。结论:子宫颈脱落细胞HPV L1壳蛋白检测在HPV阳性妇女的子宫颈病变筛查中具有一定的价值,可能成为一种合适的分流方法。     

宫颈低级别鳞状上皮内病变宫颈锥切术后病理升级的相关因素分析

目的 探讨宫颈低级别鳞状上皮内病变（LSIL）宫颈锥切术后病理升级的发生率及其相关影响因素。方法 纳入2014年1月至2022年4月于潍坊医学院附属医院因活检病理提示宫颈LSIL有指征行宫颈锥切的患者97例,分为病理升级组和病理未升级组进行分析。结果 97例患者中,54例（55.7%）宫颈锥切术后病理升级,两组一般资料、术前及术后人乳头瘤病毒（HPV）感染、宫颈液基细胞学（TCT）、阴道镜检查结果、累及腺体及锥体高度等比较,差异均无统计学意义（P>0.05）。将危险因素组合后行卡方检验显示,HPV16/18阳性伴TCT高级别鳞状上皮内病变（HSIL）/不排除高级别鳞状上皮内病变不典型鳞状细胞（ASC-H）或不典型腺上皮细胞（AGC）是宫颈锥切术后病理升级的危险因素（P=0.041）。结论 HPV16/18型阳性伴TCT>LSIL(HSIL/ASC-H/AGC)是影响活检LSIL术后病理升级的影响因素,对LSIL患者进行分流管理时需同时考虑患者术前HPV及TCT结果。LSIL宫颈锥切术后病理是否升级不影响患者术后HPV转归情况。     

人乳头瘤病毒E7蛋白检测子宫颈上皮内病变的价值

目的探讨人乳头瘤病毒(HPV)E7蛋白检测宫颈病变的临床应用价值。方法选取180例高危型HPV阳性妇女,采用免疫细胞化学方法检测宫颈脱落细胞HPV E7蛋白。结果 (1)病理高级别宫颈上皮内病变(HSIL)及浸润癌组HPV E7蛋白阳性率(77.8%,56/72)显著高于低级别宫颈上皮内病变(LSIL)组(52.1%,25/48;P=0.003),病理LSIL组阳性率显著高于宫颈炎组(26.7%,16/60;P=0.007);(2)对于细胞学为ASCUS和LSIL的妇女,HPV E7蛋白诊断HSIL及宫颈癌的灵敏度、特异度、阳性预测值及阴性预测值分别为97.1%、52.1%、58.9%和96.2%;(3)HPV E7蛋白和TCT诊断LSIL及以上病变的灵敏度、特异度、阳性预测值、阴性预测值分别为67.5%vs 51.7%、73.3%vs 58.3%、83.5%vs 71.3%、53.0%vs 37.6%,除特异度外,E7蛋白诊断的价值优于TCT(P0.05)。结论 HPV E7蛋白在检测宫颈病变,尤其是在细胞学轻度异常患者的分流中,具有一定的临床应用价值。     

HPV L1壳蛋白检测联合细胞学、HPV检测诊治宫颈病变的临床价值

目的探讨HPV L1壳蛋白检测联合细胞学检查、HPV检测在宫颈病变诊治中的临床价值。方法:收集2014年7月~2015年6月期间,在新乡医学院第一附属医院妇科门诊因体检或发现宫颈异常就诊的符合入组条件的女性1052例,进行TCT筛查、HPV分型检测,对一项或两项结果异常者进行子宫颈组织活检术,并对TCT结果异常患者采用免疫细胞化学的方法检测HPV L1壳蛋白的表达。结果 156例TCT结果异常中,意义未明确的非典型鳞状上皮细胞(ASCUS)、低度鳞状上皮内病变(LSIL)、高度鳞状上皮内病变(HSIL)、鳞状细胞癌(SCC)的HR-HPV阳性率,分别为19.1%(13/68)、35.3%(18/51)、77.8%(21/27)、100%(10/10)。各组之间比较,差异有统计学意义(P0.05)。TCT异常结果中ASCUS、LSIL、HSIL、SCC在病理组织学CIN及以上阳性率分别为54.4%(37/68),60.8%(31/51),85.2%(23/27),100%(10/10)。各组之间的比较,差异有统计学意义(P0.05)。随着组织病理学诊断病变级别越高,HPV L1壳蛋白阳性表达率逐渐下降。结论 TCT和HPV分型检测在宫颈癌筛查中必不可少。HPV L1壳蛋白检测可以预测宫颈癌前病变的进展趋势。三者联合应用可以及早发现宫颈癌前病变,并对病变进展风险进行合理的评估。     

子宫颈液基细胞学检查异常的涂片中人乳头状瘤病毒L1蛋白的表达及其意义

目的 探讨人乳头状瘤病毒(HPV)L1蛋白在官颈液基细胞学检查异常涂片中的表达及其意义.方法 选择2006年9月-2008年9月间,在中日友好医院就诊的官颈液基细胞学检查诊断为≥未明确诊断意义的不典型鳞状上皮细胞(ASCUS),且其第2代杂交捕获试验(HC-Ⅱ)榆测HPV DNA结果均为阳性,同时有组织病理学诊断的患者共274例.其中,宫颈液基细胞学检查诊断为ASCUS 105例、低度鳞状上皮内病变(LSIL)119例、不除外高度病变的不典型鳞状上皮细胞(ASC-H)9例、高度鳞状上皮内病变(HSIL)36例、官颈鳞癌5例;组织病理学检查(作为金标准)诊断为炎症96例、宫颈上皮内瘤变(CIN)Ⅰ 85例、CIN Ⅱ 55例、CIN Ⅲ 32例、官颈鳞癌6例.对此274例患者的官颈涂片,采用伞反应抗体的免疫细胞化学染色进行HPV L1蛋白的榆测,分析其对官颈病变进展的预测价值.结果 274例患者中,组织病理学检杏诊断为炎症的组织中HPV L1蛋白阳性表达率为69.8%(67/96),CIN Ⅰ为83.5%(71/85),CIN Ⅱ为41.8%(23/55),CIN Ⅲ为3.1%(1/32),宫颈鳞癌为0(0/6),除CIN Ⅲ与官颈鳞癌比较,差异无统计学意义(P>0.05)外,其他不同病变问比较,差异均有统计学意义(P<0.01);细胞学检查诊断为LSIL的细胞中HPV L1蛋白阳件表达率(75.6%,90/119)最高,其次为ASCUS细胞(63.8%,67/105)和HSIL+宫颈鳞癌细胞(9.8%,4/41),3者问比较,差异有统计学意义(P<0.01).71例未经治疗的ASCUS、LSIL患者中,55例HPV L1蛋白阳性表达者中无一例疾病进展,16例HPV L1蛋白阴性表达者疾病进展的发生率为19%(3/16),两者比较,差异有统计学意义(P<0.01).结论 HPV L1蛋白在宫颈液基细胞学检查异常涂片中的表达情况可以帮助了解宫颈的病变程度,预测宫颈病变的发展趋势,尤其对细胞学检查诊断为ASCUS和LSIL的患者可协助指导临床处理.     

HPV E6/E7 mRNA、p16/Ki67检测在ASCUS中的诊断价值

目的:探讨HPV E6/E7 mRNA、p16/Ki67检测在宫颈细胞学检查为意义不明确的不典型鳞状细胞(ASCUS)中的诊断价值。方法:回顾分析2015年12月至2017年5月在郑州大学第三附属医院就诊的液基薄层细胞学检查(TCT)结果为ASCUS,并行阴道镜下宫颈组织活检的患者200例。应用杂交捕获技术(HC2)和支链DNA技术(b DNA)行HPV DNA、HPV E6/E7 mRNA检测。免疫组化法检测宫颈组织中p16/Ki67表达。结果:宫颈高级别病变组(包括CIN2,CIN3,宫颈癌,简称CIN2+)中p16/Ki67、HPV E6/E7mRNA的阳性率与炎症/CIN1组比较,差异有统计学意义(χ2=31. 952,P=0. 000;χ2=11.231,P=0.001),且p16/Ki67表达与CIN2+具有一致性(kappa=0.400,P=0.000)。炎症/CIN1组中,HPV E6/E7 mRNA检测与p16/Ki67检测结果间的差异有统计学意义(P=0.000),但在CIN2+中两者差异无统计学意义(P=0.375)。ROC曲线分析p16/Ki67检测、HPV E6/E7 mRNA检测诊断CIN2+的准确性分别为(AUC=0.800,0.625),均高于HPV DNA检测(AUC=0.579)。结论:HPV E6/E7 mRNA、p16/Ki67表达与宫颈高级别病变密切相关,HPV E6/E7 mRNA检测可望代替HPV DNA成为分流ASCUS的一种有效手段,而p16/Ki67与宫颈高级别病变显著一致,可辅助用于ASCUS患者宫颈组织的病理诊断。     

妊娠期妇女子宫颈细胞学检查结果异常的相关因素分析

目的 探讨妊娠期妇女宫颈细胞学检查结果异常的相关因素.方法 选择2007年9月至2008年9月在首都医科大学附属北京妇产医院产科定期产前检查的妊娠12～36周的妇女,除外先兆流产、胎膜早破、前置胎盘等并发症共12 112例,于初次产前检查时常规行宫颈液基细胞学检查(TCT),同时记录妊娠期妇女的职业、受教育程度、户口地址、家庭收入、民族、初次性生活年龄、性伴侣个数、避孕方法、孕产史、妇科伴随症状、相关肿瘤家族史、既往妇科病史、吸烟史,并记录当天妇科检查情况,分析TCT结果异常的相关危险因素.结果 资料完整的妊娠期妇女共11 906例,资料满意率为98.30%(11 906/12 112).其中,TCT结果正常10 354例,占86.96%(10 354/11 906)、未明确诊断意义的不典型鳞状上皮细胞(ASCUS)1134例,占9.52%(1134/11 906)、未明确诊断意义的不典型腺上皮细胞(ACUS)112例,占0.94%(112/11 906)、低度鳞状上皮内病变(LSIL)229例,占1.92%(229/11 906)、高度鳞状上皮内病变(HSIL)74例,占0.62%(74/11 906),宫颈鳞癌3例,占0.02%(3/11 906).多因素非条件logistic回归分析结果显示,与ASCUS及AGUS相关的危险因素包括初次性生活年龄(OR_(ASCUS)=2.90、OR_(AGUS)=7.32)、性伴侣个数(OR_(ASCUS)=1.49、OR_(AGUS)=2.02)、流产次数(OR_(ASCUS)=1.68、OR_(AGUS)=3.50);与LSIL、HSIL相关的危险因素包括初次性生活年龄(OR_(LSIL)=6.34、OR_(HSIL)=9.26)、性伴侣个数(OR_(LSIL)=1.69、OR_(HSIL)=1.65)、流产次数(OR_(LSIL)=1.53、OR_(HSIL)=5.33)、吸烟(OR_(LSIL)=1.84、OR_(HSIL)=1.77);与TCT结果异常(包括ASCUS、AGUS、LSIL、HSIL)相关的感染因素包括滴虫性阴道炎(P<0.01)、人乳头状瘤病毒(HPV)感染(P<0.01);柱状上皮异位与TCT结果异常密切相关(χ~2=43.269,P=0.000),但与柱状上皮异位的程度无关.结论 妊娠期妇女宫颈细胞学检查结果异常发生的相关因素与非妊娠期相同.     

细胞块技术检测宫颈脱落细胞p16INK4A表达对评价宫颈病变的意义

目的探讨将细胞块技术应用于宫颈细胞学诊断并检测宫颈脱落细胞p16INK4A表达,对评价宫颈病变的意义.方法采用链霉素抗生物-过氧化物酶(SP)法,检测79例细胞块上的宫颈脱落细胞及相应的宫颈活检组织中p16INK4A表达.结果在宫颈脱落细胞中,p16INK4A在正常、未明确意义的不典型鳞状细胞(ASCUS)、低度鳞状上皮内病变(LSIL)、高度鳞状上皮内病变(HSIL)、鳞状细胞癌(SCC)阳性表达率分别为0%、18.18%、46.43%、89.66%、100%.在相应活检组织中,p16INK4A在宫颈炎、CINⅠ、CINⅡ、CINⅢ、SCC中阳性表达率分别为0%、61.90%、87.50%、100%、100%.p16INK4A在宫颈脱落细胞和组织中表达的符合率为87.34%,宫颈脱落细胞p16INK4A免疫组化染色使检出宫颈高度病变的敏感性提高了21.95%.结论细胞块技术可用于宫颈细胞学诊断,检测宫颈脱落细胞p16INK4A表达可以提高宫颈高度病变的检出率,减少漏诊率.     

Role of Cervical Cancer Biomarkers p16 and Ki67 in Abnormal Cervical Cytological Smear

IntroductionCervical cancer is the most common cancer in India. Screening for cervical cancer helps in marked reduction of invasive cervical cancers. The low sensitivity of Papanicolaou cytology (Pap smear) and high-risk human papillomavirus (HR-HPV) in excluding high-grade intraepithelial lesion (ASC-H) leads to unnecessary referrals to colposcopy-guided biopsy. The combined cervical cytology screening and HR-HPV have its own limitations and still need further standardization. Using additional biomarkers like staining with p16 and Ki-67 might help in triaging abnormal pap smear.Materials and MethodsA prospective, cross-sectional study was performed over a period of 16 months in the Department of Obstetrics and Gynaecology, in collaboration with Department of Pathology. Study was conducted to know the efficacy of immunostaining with p16/Ki-67 in predicting the presence of significant lesion in cases of mild cytological atypia. PAP smears (conventional and LBC) along with P16, Ki-67 and available biopsies were correlated.ResultsLiquid-based cytology (LBC) was done in 2134 cases, out of which 46 cases showed abnormal cytological findings such as [22 atypical squamous cells of undetermined significance (ASCUS), 3 low-grade squamous intraepithelial lesion (LSIL), 8 atypical squamous cells cannot exclude high-grade lesion (ASC-H), 6 high-grade squamous intraepithelial lesion (HSIL), 5 squamous cell carcinoma (SCC), 2 adenocarcinoma, 1 atypical glandular cells of undetermined significance (AGUS)]. Immunostaining with p16 and Ki-67 was performed on 38 cases of abnormal cytological smears. Out of 38 abnormal cytology cases, 28 cases had shown co-staining for both p16 and Ki-67, suggestive of true HPV infection of the cells. Of the 38 cases, 07/14 ASCUS, 06/06 HSIL, 07/08 ASC-H, 05/05 squamous cell carcinoma and 02/02 adenocarcinoma also showed dual positivity for p16 and Ki-67. One case of AGUS was diagnosed, but the smear was unsatisfactory for immunocytochemical evaluation and excluded from the study. Three cases of LSIL were also diagnosed on cytological evaluation, and 1 of them however showed positivity for p16 and Ki-67 on immunocytochemistry (ICC). In the ASC-US group, the sensitivity and specificity of the immunostaining in diagnosing CIN2 + lesions were 87.51%, and in LSIL group, the sensitivity and specificity of the immunostaining in diagnosing CIN2 + lesions were 100%. p16/Ki-67 positivity also increased with cytological severity which in turn corresponded with histological findings: it reached from 50% in ASC-US to 100% in both HSIL and SCC categories.ConclusionThis immunostaining with p16 and Ki67 can be a useful method in the triaging of the ASC-US and the LSIL group as considering the high sensitivity and specificity values.     

p16/Ki-67双染在宫颈癌及癌前病变初筛中的应用价值研究

目的:评价采用p16/Ki-67双染检测技术作为宫颈癌及癌前病变初筛方法的效果及应用价值。方法:对重庆市万州区982例年龄35~64岁有性生活的妇女进行宫颈癌筛查。每位妇女均接受了人乳头瘤病毒DNA(HPV DNA)检测、Thin Prep液基细胞学检查、p16/Ki-67双染检测,对结果异常者进行阴道镜检查,阴道镜下在可见病变处直接取活检,无可见病变时,行宫颈管搔刮术(ECC)。比较3种方法分别作为初筛手段识别宫颈癌前病变(高级别鳞状上皮内病变)及浸润癌的灵敏度、特异度、阳性预测值和阴性预测值等指标,以受试者工作特征(ROC)曲线下面积(AUC)综合分析3种方法作为宫颈癌初筛手段的应用价值。结果:最终966例妇女进入研究,共检出高级别鳞状上皮内病变及浸润癌患者42例。HPV DNA检测、液基细胞学检查和p16/Ki-67检测对宫颈癌及癌前病变患者的灵敏度分别为97.6%、88.1%、92.9%;特异度分别为84.1%、78.8%、82.8%;阳性预测值分别为21.8%、15.9%、19.7%;阴性预测值分别为99.9%、99.3%、99.6%。p16/Ki-67检测的AUC分别与HPV DNA检测、液基细胞学检查相比,差异均无统计学意义(P0.05)。结论:p16/Ki-67双染检测初筛宫颈癌及癌前病变的效果与HPV DNA检测及液基细胞学检查相似,因其具有简便、客观、高效、易于重复的特点,p16/Ki-67双染检测为宫颈癌及癌前病变的有效初筛提供了一种新选择。     

宫颈薄层细胞学中应用细胞块及Ki-67免疫组化染色技术的价值分析

目的:探讨在宫颈薄层细胞学中应用细胞块及免疫组化染色技术评价宫颈病变的意义。方法:采用免疫组化SP法,检测79例患者宫颈脱落细胞及其宫颈活检组织中K i-67的表达。结果:在细胞块上检测宫颈脱落细胞K i-67的表达,正常、ASCUS、LSIL、HSIL、SCC阳性表达率分别为0%(0/2)、27.27%(3/11)、57.14%(16/28)、93.10%(27/29)、100%(9/9)。K i-67免疫组化染色检出宫颈高度病变的敏感性、特异性、阳性预测值、阴性预测值分别为90.24%、76.31%、80.43%、93.54%。结论:宫颈薄层细胞学中应用细胞块及免疫组化染色技术可以提高宫颈高度病变的检出率。     

宫颈鳞状上皮内瘤变二级命名法的临床应用

目的探讨2013年WHO关于宫颈鳞状上皮内瘤变二级命名法的临床应用及相关问题。方法采用免疫组化技术检测p16、Ki-67蛋白在111例宫颈良性反应性病变（良性病变组）、81例CIN1、39例CIN2患者组织中的表达情况。并分析宫颈低级别及高级别鳞状上皮内瘤变二级命名法中,不同级别病变与p16、Ki-67蛋白表达的关系。结果 p16在良性病变、CIN1和CIN2组中的阳性表达率分别为18.92%、41.98%和64.10%,良性病变组与CIN1、CIN2组比较,差异均有统计学意义（P〈0.05）;Ki-67在良性病变、CIN1和CIN2组中的阳性表达率分别为32.43%、32.10%和69.23%,CIN2组与CIN1组、良性病变组比较,差异均有统计学意义（P均〈0.05）。p16和Ki-67蛋白联合检测：在良性病变、CIN1、CIN2组中,p16（＋）/Ki-67（＋）的表达率分别为11.71%、9.88%和43.59%,p16（＋）/Ki-67（-）的表达率分别为7.21%、32.10%和20.51%;p16（-）/Ki-67（＋）的表达率分别为20.72%、22.22%和25.64%;p16（-）/Ki-67（-）的表达率分别为60.36%、35.80%和10.26%。三组中p16阴性着色特点不同,Ki-67的阳性着色特点不同。结论 p16蛋白联合Ki-67可以很好地区分低级别和高级别宫颈鳞状上皮内瘤变,但其有赖于免疫组化判读标准的严格执行;同一级别不同的染色特点可能代表组织的不同生物学特性,但尚需大量的临床验证。     

The expression of p16 (INK4a) and Ki-67 in relation to high-risk human papilloma viral load and residual disease after conization with positive margins

The purpose of this study was to investigate the correlations between high-risk human papillomavirus (HPV) load and p16 (INK4a) or Ki-67, and to identify biomarkers that may predict residual disease after conization with positive margins. The following samples were analyzed: 49 paraffin-embedded specimens from patients with cervical intraepithelial neoplasia (CIN), including 12 CIN 2 conization specimens and 37 CIN 3 conization specimens. Immunohistochemical analysis was performed with antibodies to p16 (INK4a) and Ki-67. Hybrid Capture II testing was used to detect high-risk HPV DNA. The mean HPV loads within each of the p16 (INK4a)-staining cases were 9.5 (relative light units/positive control) RLU/PC for negative staining, 531.8 RLU/PC for 1+ staining, 140.2 RLU/PC for 2+ staining, and 545.1 RLU/PC for 3+ staining. HPV loads differed significantly according to p16 (INK4a) expression (P = 0.0021). The mean HPV loads within Ki-67 staining cases were 28.2 RLU/PC for 1+ staining, 189.6 RLU/PC for 2+ staining, and 563.3 RLU/PC for 3+ staining. HPV loads differed significantly according to Ki-67 expression (P = 0.0259). The expression of p16 (INK4a) (P = 0.0012) and Ki-67 (P = 0.0006) were significantly associated with the CIN grade. In univariate and multiple logistic regression analysis, age, parity, cytology, lesion grade in the cone, high-risk HPV load, and the expression of p16 (INK4a) and Ki-67 were not significantly associated with residual lesions after conization with positive margins (P > 0.05). In conclusion, high-risk HPV load showed significant differences according to the expression of p16 (INK4a) and Ki-67, while none of the prognostic factors were significantly associated with residual disease after conization with positive margins.     

p16/Ki-67 dual staining in cervico-vaginal cytology: correlation with histology, Human Papillomavirus detection and genotyping in women undergoing colposcopy

Objectives

To evaluate the CINtec® PLUS assay (mtm laboratories), a new immunocytochemical method for the simultaneous detection of p16^INK4a and Ki-67, in liquid-based cervico-vaginal cytology, investigating the association of the dual staining with HPV infection and genotyping as well as cytological and histological abnormalities.

Methods

140 women with a cervico-vaginal sample obtained immediately before the colposcopy were enrolled. This cytological sample was used for HPV testing with the Linear Array HPV Genotyping Test, the dual staining with the CINtec® PLUS kit and the morphology assessment.

Results

Cytology results were 38 NILM, 16 ASC-US, 32L-SIL, 54H-SIL or worse. 113 patients also had a colposcopy-guided biopsy, classified as 14 negative, 35 CIN1, 24 CIN2, 37 CIN3, 3 invasive SCC. A strong association between p16/Ki-67 and HR-HPV infection was found (COR = 6.86, 95% CI: 1.84-31.14). Importantly, the association between p16/Ki-67 positivity and HPV16 and/or 18 infection was 2-fold stronger compared to that with the infection by other HR-HPV types (COR = 9.92, 95% CI: 2.39-47.77 vs COR = 4.20, 95% CI: 0.99-20.87). In addition, p16/Ki-67 positivity rate significantly increased with the severity of the cytological and histological abnormalities (p < 0.05 in both cases). p16/Ki-67 positivity resulted strongly associated with a CIN2 + diagnosis (COR = 10.86 95% CI: 4.16-29.12).

Conclusions

This preliminary study evidenced that p16/Ki-67 immunostaining might have a relevant clinical role, since the dual staining was significantly associated with HR-HPV infection, particularly with HPV 16 and 18, and the increasing grade of the cervical lesions, the positivity for this biomarker being strongly related to the presence of a CIN2 + lesion.     

Ki-67、p63、bcl-2蛋白在葡萄胎妊娠与水肿性流产中的诊断价值

目的:研究Bcl-2、p63和Ki-67蛋白在鉴别完全性葡萄胎(CHM)、部分性葡萄胎(PHM)、水肿性流产(HA)中的应用价值。方法:收集山东大学齐鲁医院病理科2012年1月至2018年6月的妊娠早期葡萄胎(HM)与水肿性流产样本共139例。对以上病例样本重新阅片,同时结合p57免疫组化染色,筛选出CHM 55例,PHM 29例,HA 40例。对筛选出的样本行Bcl-2、p63、Ki-67免疫组化染色。结果:Ki-67用于鉴别HA与PHM的敏感度、特异性、阳性预测值(PPV)、阴性预测值(NPV)分别为82.5%、72.4%、80.5%和75.0%;p63用于鉴别HA与PHM的敏感度、特异性、PPV、NPV分别为95.0%、79.3%、86.4%和92.0%;p63联合Ki-67用于鉴别HA与PHM的敏感度、特异性、PPV、NPV分别为95.0%、79.3%、86.4%和92.0%。p63联合Ki-67用于鉴别HA与PHM的受试者工作特征(ROC)曲线下面积(0.934)显著高于p63(0.898),差异有统计学意义(P<0.05)。结论:Ki-67与p63用于鉴别PHM与HA具有一定的应用价值,联合使用Ki-67与p63鉴别PHM与HA比单独使用两者的准确性更高。     

Progression of cervical low grade squamous intraepithelial lesions: in search of prognostic biomarkers

Objective

It has been reported that approximately 10% of low grade squamous intraepithelial lesions (LSIL) progress to high grade squamous intraepithelial lesions (HSIL) within a 2-year follow up. The factors related to lesion progression are currently unknown. The aim of the study was to identify prognostic markers of the course of LSIL. This retrospective study was designed to correlate regression, persistence and progression of biopsy-proven LSIL with patients’ age, HPV genotypes and immunohistochemical expression of the main cell cycle regulating proteins: p53, pRb, p16, and Ki-67.

Study design

A total of 584 consecutive patients with biopsy proven LSIL and 2-year follow-up were included in the age analysis. HPV genotyping was performed in 328 LSIL cases using the SPF10 PCR-LiPA25 (version 1), 238 LSIL cases were immunostained for Ki-67 and p16, and 101 cases were immunostained for pRb and p53.

Results

The odds of LSIL persistence and progression were significantly higher in women 30–39, 40–49 and 50+ years old, as compared to women 20–29 years old (OR 1.89, 2.52 and 2.39, respectively). The odds of persistence and progression were higher in women infected with HPV16, 18, 33 and 52 (OR 3.5, 3.1, 3.5 and 2.9, respectively). There were no significant differences in expression of immunomarkers (p16, p53, pRB and Ki-67) between the lesions that regressed versus the lesions that persisted or progressed.

Conclusions

Patients 30 years of age and older have a higher risk of LSIL progression or persistence as compared to 20–29 year olds. In addition, HPV genotyping, but not the cell cycle markers, may aid in prognosis of LSIL course.     

Expression of p53, Ki-67, and CD31 in the vaginal margins of radical hysterectomy in patients with stage IB carcinoma of the cervix

OBJECTIVES: This study was undertaken to evaluate the expression of p53, Ki-67, and CD31 both in the tumor and in the vaginal margins of radical hysterectomy in patients with stage IB squamous cell carcinoma of the cervix, as an attempt to use these proteins as possible markers for residual tumor in cervical cancer. METHODS: Thirty patients with stage IB squamous cell carcinoma of the cervix were submitted to radical hysterectomy (study group), and thirty patients with uterine myoma were submitted to vaginal hysterectomy (control group) and were prospectively studied from November 2001 to September 2002. Tissue samples were taken from the tumor or cervix, anterior vaginal margin (AVM), and posterior vaginal margin (PVM) and were immunohistochemically evaluated by monoclonal antibodies for p53, Ki-67, and CD31. Vaginal samples in which the histological examination showed tumor involvement were excluded from the study. RESULTS: Patient's mean age was 48.7 +/- 10.4 years (27-73 years). The clinical stage was IB1 in 22 patients (73.3%) and IB2 in eight patients (26.7%). The expressions of p53, Ki-67, and CD31 were significantly higher in the tumor than in the benign cervix (P < 0.001). Higher expressions of these markers were noted in the vaginal margins of radical hysterectomy in patients with cervical carcinoma compared to the vaginal margins of control patients. This association was demonstrated for p53 in the AVM proximal (P = 0.045), for Ki-67 in AVM proximal (P < 0.001), AVM distal (P < 0.001), PVM proximal (P = 0.009), and PVM distal (P < 0.001), and for CD31 in AVM proximal (P = 0.003) and AVM distal (P = 0.018). There was no difference in p53, Ki-67, and CD31 expression between the proximal and distal regions of the vaginal margins in patients with carcinoma of the cervix. CONCLUSION: The expressions of p53, Ki-67, and CD31 were significantly higher in both the histologically positive (cervical tumor) and negative (vaginal margins) tissues of patients who had undergone radical hysterectomy for cervical cancer compared to the benign control tissues.     

Triaging HPV-positive women with p16/Ki-67 dual-stained cytology: Results from a sub-study nested into the ATHENA trial

Objectives

In addition to genotyping for HPV16/18, dual-immunostaining for p16/Ki-67 has shown promise as a triage of HPV-positive women. We assessed the performance of p16/Ki-67 dual-stained cytology for triaging HPV-positive women undergoing primary HPV screening.