Demonstration of silver-stained nucleolar organizer region associated proteins (AgNORs) after wet autoclave pretreatment in breast carcinoma: Correlation to tumor stage and long-term survival

Argyrophilic nucleolar organizer region associated proteins (AgNORs) are known to reflect cellular and nucleolar activity. Due to a novel staining procedure, which substantially improves visualisation of AgNORs on formalin-fixed and paraffin-embedded material, AgNORs can be reliably demonstrated as true substructures of the nucleoli. The aim of the present study was to apply a standardized morphometric AgNOR quantification on a large series of breast carcinomas with regard to its prognostic relevance. AgNOR quantity was evaluated on archival tumor tissues of 115 adenocarcinomas of the breast treated with the wet autoclave method prior to standardized silver-staining and morphometric analysis. AgNOR parameters were correlated to prognostic features (steroid hormonal receptor status, tumor type, tumor size, histological grading, pTNM, and UICC stage) carrying out both univariate and multivariate survival analyses. AgNOR number and area were proven to be statistically significantly related (Pearson correlation coefficient: 0.67, Bonferroni adjusted P = 0.0001). Almost all AgNOR parameters, in particular CV (coefficient of variation) of corrected area (-area) and CV of number, were statistically significantly correlated to estrogen and progesterone receptor status as well as histological grading of tumors. Increased AgNOR parameters were statistically significantly associated with early tumor relapse and cancer related death. Univariate and multivariate analysis by means of Cox regression revealed independent prognostic significance for CV of -area and number of AgNORs. Various AgNOR parameters (CV of number, CV of -area, CV of area, mean -area, and mean area of AgNORs per nucleus) determined on wet autoclave pre-treated formalin-fixed and paraffin-embedded breast cancer tissues are statistically highly significantly associated with the prognostic outcome, independently predicting tumor-free and overall survival.     

Significance of immunohistochemistry in breast cancer

The biological characteristics of the tumour are used to estimate prognosis and select appropriate systemic therapy for patients with (breast) cancer. The advent of molecular technology has incorporated new biomarkers along with immunohistochemical and serum biomarkers. Immunohistochemical markers are often used to guide treatment decisions, to classify breast cancer into subtypes that are biologically distinct and behave differently, and both as prognostic and predictive factors. Steroid hormone receptors, markers of tumour proliferation, and factors involved in angiogenesis and apoptosis are of scientific interest. In this review we will provide information on the immunohistochemical markers used in the management of breast cancer patients using available data from the literature. We consider the utility of established immunohistochemical markers, and discuss the challenges involved in integrating novel molecular markers into clinical practice.     

前列腺癌AgNOR计数与DNA含量测定的意义

作者应用银染色技术对３０例前列腺癌、２０例前列腺增生症组织中的核仁组成区相关嗜银蛋白（ＡｇＮＯＲ）进行检测，并采用图像分析技术测定其ＤＮＡ含量。结果显示，在前列腺癌组织中，ＡｇＮＯＲ计数与ＤＮＡ含量均明显高于前列腺增生症组织（Ｐ＜０．００１）；肿瘤分化越差，ＡｇＮＯＲ计数和ＤＮＡ含量越高，并且与患者预后相关（Ｐ＜０．００１）；ＡｇＮＯＲ计数与ＤＮＡ含量呈明显的正相关（Ｐ＜０．０１）。结果提示ＡｇＮＯＲ计数与ＤＮＡ含量可作为反映前列腺癌生物学行为的指标。     

AgNOR变化在移行上皮肿瘤中的诊断意义

应用 AgNOR 染色技术对32例泌尿系移行上皮及其肿瘤组织细胞核内 AgNOR 的定量研究表明,正常移行上皮与移行上皮乳头状瘤之间 AgNOR 差异无显著性(P>0.05),而移行上皮乳头状瘤、不同恶性程度的移行上皮癌各组间 AgNOR 比较,差异均有显著性(0.01相似文献   

Practical applications of immunohistochemistry in the diagnosis of soft tissue neoplasms

Immunohistochemistry is an extraordinarily powerful tool in the armamentarium of the diagnostic surgical pathologist. This article discusses some ways in which immunohistochemistry assists in the diagnosis of soft tissue neoplasms. Four common clinicopathologic scenarios in soft tissue pathology are presented and a practical approach to these rare neoplasms is discussed. The limitations of immunohistochemistry and possible future directions for the application of immunohistochemistry to sarcoma diagnosis are also reviewed.     

AgNOR检测对宫颈癌预后的意义

 对73例随访10年以上的宫颈鳞癌进行A_gNOR计数分析，发现组织学D级的肿瘤细胞核内A_gNOR均值高于组织学Ⅰ级者为5.53± 1.47及4.40±1.34)。10年生存率亦有差异性，A_gNOR均值≥5者10年生存率为18%，而<5的10年生存率为46%(P<0.02)。结果表明，A_gNOR计数与宫颈鳞癌的组织学分级及临床预后有关。本文认为A_gNOR计数可作为一种肿瘤定量指标，对判定肿瘤恶性程度，预测宫颈癌预后具有一定参考价值。     

Utility of immunohistochemistry in breast cancer practice

Immunohistochemistry (IHC) has been used to search for specific protein-products, to observe the unique characteristics of breast cancer, to differentiate breast cancer from cancers of other sites origin, to obtain information regarding genetic alterations, to investigate prognostic factors, and to provide predictive factors for efficient treatments. Technical developments make it possible to utilize IHC as a convenient diagnostic tool, and to use it in advanced protocols in combination with other histochemical techniques as well. In this review article, the development of IHC, IHC for differential diagnosis and IHC for providing information about hormone receptors and HER2 in breast cancer practice will be introduced. Standardization and quality control are critical issues to utilize IHC adequately in breast cancer practice.     

Value of liver scintigraphy in pretreatment staging and in follow-up of patients with malignant melanoma

Liver scintigraphy (99Tcm sulphur colloid) was performed in 118 patients with malignant melanoma. In 73 patients diagnosed as stage I, the pretreatment evaluation showed one false-negative and one false-positive examination. During follow-up there were ten abnormal liver scintigraphies; one was later correlated to liver metastases. In 46 patients diagnosed as stage II-IV, the pretreatment liver scintigraphy yielded false-negative results in 36% and false-positive results in 15%. The predictive values of positive and negative tests were 44% and 81%, respectively. The yields of liver tests (S-alkaline phosphatase, S-gamma-glutamyl-transferase) in patients with liver metastases were low. This study demonstrated the limitations of liver scintigraphy for diagnosis of liver metastases in patients with malignant melanoma.     

乳腺癌细胞的DNA含量、AgNOR计数及与预后的关系

 本文采用流式细胞术检测了54例乳腺肿瘤细胞的DI、PI和S期细胞比例等项参数，其中34例乳腺癌进行了A_gNOR计数和5年～14年随访。经分析表明：DI、PI和S期细胞比例可区分肿瘤的良恶性，A_gNOR计数、DI、PI和S期细胞比例与乳腺癌的预后有关。     

Comparative study of outcome of type I tympanoplasty in dry and wet ear

Chronic suppurative otitis media is a common condition seen in patients attending the otolaryngology clinic. The discharging ear presents the otologist with the dilemma of operating on it or not. This due to the widespread belief that the success rate while doing tympanoplasty on wet ears is decidedly inferior. To evaluate this fact we conducted a study to compare outcomes of type 1 tympanoplasty in dry and wet ears. Wet ear meant that the patient had a mild mucoid discharge which was negative on culture. Type 1 tympanoplasty was done in all patients under local anesthesia using temporalis fascia graft and by underlay technique. We conclude that the presence of discharge in the ear at the time of operation does not interfere with the results of tympanoplasty, but it should be mucoid and scanty.     

免疫组织化学在淋巴组织增生性病变病理诊断中的应用

免疫组织化学是淋巴组织增生性病变病理诊断与鉴别诊断不可缺少的方法之一,但是必须正确应用和评估其免疫表型结果.重点应注意下列几点:(1)同类抗体的合理配伍;(2)免疫表型假阳性与假阴性的识别;(3)免疫表型阳性定位正确性的识别;(4)抗体非特异性的识别;(5)瘤细胞与反应性细胞免疫表型的识别;(6)免疫表型在淋巴组织增生性病变良恶性鉴别诊断中的正确应用.强调免疫组织化学技术须与其他淋巴组织增生性病变诊断要素相结合才能获得满意的结果.     

Prognostic Significance of Two Dimensional AgNOR Evaluation in Local Advanced Rectal Cancer Treated with Chemoradiotherapy

The prognostic significance of AgNOR proteins in stage II-III rectal cancers treated with chemoradiotherapy was evaluated. Silver staining was applied to the 3μm sections of parafin blocked tissues from 30 rectal cancer patients who received 5-FU based chemoradiotherapy from May 2003 to June 2006. The microscopic displays of the cells were transferred into the computer via a video camera. AgNOR area (nucleolus organizer region area) and nucleus area values were determined as a nucleolus organizer regions area/total nucleus area (NORa/ TNa). The mean NORa/TNa value was found to be 9.02±3.68. The overall survival and disease free survival in the high NORa/TNa (>9.02) patients were 52.2 months and 39.4 months respectively, as compared to 100.7 months and 98.4 months in the low NORa/TNa (<9.02) cases. (p<0.001 and p<0.001 respectively). In addition, the prognosis in the high NORa/TNa patients was worse than low NORa/TNa patients (p<0.05). In terms of overall survival and disease-free survival, a statistically significant negative correlation was found with the value of NORa/TNa in the correlations tests. Cox regression analyses demostrated that overall survival and disease-free survival were associated with lymph node status (negative or positive) and the NORa/TNa value. We suggest that two-dimensional AgNOR evaluation may be a safe and usable parameter for prognosis and an indicator of cell proliferation instead of AgNOR dots.     

免疫组化在胸膜上皮性恶性肿瘤鉴别诊断中的价值

背景与目的 胸膜上皮性恶性肿瘤鉴别诊断难度较大,本文旨在探讨免疫组化在胸膜恶性间皮瘤与转移性肺腺癌鉴别诊断中的作用.方法 用免疫组化S-P法检测56例胸膜上皮性恶性肿瘤中波形蛋白、间皮细胞(MC)、钙结合蛋白(CR)、甲状腺转录因子-1(TTF-1)、癌胚抗原(CEA)、表面活性蛋白-B(Sp-B)、细胞角蛋白(CK)的表达情况.结果 用免疫组化方法确定56例胸膜上皮性恶性肿瘤中恶性间皮瘤24例,转移性肺腺癌22例.波形蛋白、MC、CR、TTF-1、CEA、Sp-B在恶性间皮瘤与转移性肺腺癌中表达差异具有显著性(P＜0.001或P＜0.002).结论 用免疫组化方法鉴别胸膜恶性间皮瘤与转移性肺腺癌,波形蛋白、MC、CR、TTF-1、CEA、Sp-B是较为理想的标志物.     

10例肛管直肠恶性黑色素瘤临床病理免疫组化研究

目的研究肛管直肠恶性黑色素瘤（简称恶黑）的临床病理特点及免疫组化染色在恶黑诊断中的作用。方法对肛管直肠恶黑的临床资料进行回顾性分析，用免疫组化s—P法作HMB45、S-100、Vimetin等染色。结果10例肛管直肠恶黑临床初步诊断恶黑1例，误诊其它疾病9例。病理形态：上皮样细胞为主7例，梭形细胞为主2例，小细胞似淋巴细胞样细胞为主1例。免疫组化染色：10例HMB-45、s-100均阳性，9例Vimetin阳性，其中1例CK阳性，LCA阴性。结论肛管直肠恶黑临床表现大便带鲜血，无明显黏液，肛门异物及息肉样突出肛门为特征，临床极易误诊。形态观察支持恶黑起源于表皮基底层黑色素细胞，免疫标记提示黑色素细胞起源于神经嵴。HMB45、S-100、Vimetin，三者联合应用能提高恶黑病理诊断的准确性。     

免疫组织化学在乳腺癌分子分型中的作用及目前存在的问题

目前，被公认的乳腺癌分子分型主要包括4型：腔面A型、腔面B型、人类表皮生长因子受体2（human epidermal growth factor receptor 2，HER2）过表达型和基底样型。由于基因表达谱检测常规应用受到限制，在日常的病理学诊断中，免疫组织化学替代分型在全世界范围内被广泛认可和应用。对当前免疫组织化学在乳腺癌分子分型中的作用及存在的问题进行了讨论，旨在进一步规范及促进免疫组织化学的应用。     

AgNOR计数对宫颈良恶性病变的鉴别诊断意义

目的 :探讨核仁形成区嗜银蛋白 (AgNOR)计数对宫颈良恶性病变鉴别诊断价值。方法 :采用AgNOR银染法对比 15例宫颈鳞癌与 2 2例慢性宫颈炎的AgNOR计数。结果 :宫颈鳞癌与宫颈良性病变的AgNOR计数差异有显著性 (P <0 0 1) ,宫颈不典型增生AgNOR计数也较宫颈良性增生上皮增高(P <0 0 5)。结论 :宫颈良恶性病变的AgNOR计数明显不同 ,此方法简便易行 ,可用于宫颈良恶性病变的鉴别。     

免疫组织化学标记在子宫内膜间质肉瘤诊断和治疗中的价值

 目的 探讨免疫组织化学标记在子宫内膜间质肉瘤（ESS）中的表达及其在诊断和治疗中的价值。方法 用免疫组织化学EnVision二步法对15例原发及3例转移复发ESS进行CD10、SM-MHC、h-caldesmon、AE1／3、CD99、Ki-67、CD34、c-kit、ER、PR的检测，并与其病理组织形态、临床并发症、鉴别诊断及预后对照。结果 17例CD10阳性，其中13例强阳性，7例伴平滑肌分化，3例伴上皮样分化，7例伴性索样分化；ER13例阳性，PR16例阳性； Ki-67 36 %～78 %。结论 子宫内膜间质肿瘤可伴多方向分化，主要为平滑肌和性索样分化；CD10是诊断子宫间质肿瘤的特异抗体，结合组织形态及SM-MHC、h-caldesmon等标记可增加其特异性；ER、PR常规检测主要用于指导孕激素辅助治疗。     

乳腺癌患者外周血T淋巴细胞AgNOR活性表达及其临床意义

目的:探讨乳腺癌患者外周血T淋巴细胞AgNOR的活性表达及其与临床病理因素的相关性.方法:利用KL型图像分析系统检测80例乳腺癌患者,40例乳腺纤维腺瘤患者,25例乳腺炎症患者和75例健康人外周血T淋巴细胞AgNOR的活性表达(T淋巴细胞核仁银染面积与细胞核面积比值,即I.S%),并与乳腺癌患者临床病理因素进行相关性分析.双抗体夹心法检测乳腺癌患者CA153和CEA.结果:发现乳腺癌患者外周血T淋巴细胞AgNOR的活性表达(5.88±6.05%)明显低于健康人(7.07±0.81%)、乳腺纤维腺瘤患者(6.58±0.72%)和乳腺炎症患者(9.58±0.68%)(P=0.000);乳腺炎症患者AgNOR的活性表达明显高于健康人、乳腺纤维腺瘤和乳腺癌患者(P=0.000);健康人与乳腺纤维腺瘤患者之间比较无显著性差异(P＞0.05).乳腺癌患者AgNOR阳性率(65.00%)明显优于CA153(17.50%)和CEA(12.50%)的阳性率(P=0.000).AgNOR表达与乳腺癌临床分期有关(P＜0.05),而与年龄、肿瘤大小、淋巴结转移状况无相关性(P＞0.05).结论:AgNOR阳性表达可作为评价乳腺癌发生、发展的客观生物学指标;在乳腺癌诊断、鉴别诊断和病情监测等方面有重要意义.     

骨科几种恶性肿瘤热休克蛋白表达免疫组化初步观察

目的 对骨科几种常见的恶性肿瘤进行HSPs免疫组化观察，以确定热休克蛋白在骨科肿瘤中的表达特点。方法 随机抽取33例骨科恶性肿瘤手术后病理标本石蜡块，行抗HSP60、70、90d、grp94免疫组化染色。采用半定量法评分，对四种热休克蛋白表达强度进行观察.结果 HSP60、grp94阳性表达例数多，表达强度大；HSP90表达也比较广泛，但一般以低一中强度表达为主，在骨肉瘤巾有中一高强度的表达；HSP70仅在部分肿瘤中有表达，但表达低强度为主，表达不均匀。结论 不同亚型的热休克蛋白存骨科恶性肿瘤中有不同强度的表达。     

膀胱癌变过程中PCNA表达的研究

为了解膀胱癌变过程中细胞增殖状态及其与肿瘤生物学行为、复发的关系，作者采用免疫组化方法检测了正常上皮、增生上皮、乳头状瘤和癌组织中的增殖细胞核抗原表达。增殖细胞核抗原指数在四种组织中呈增高趋势，并且与肿瘤病理分级、分期密切相关，但与肿瘤大小、数目无相关。复发者的增殖细胞核抗原指数显著高于无复发者。结果提示，膀胱癌变过程中细胞增殖能力不断增强，增殖细胞核抗原指数可以作为肿瘤生物学行为、判断肿瘤复发的一项有价值的标记物。