肝细胞涂片在评估图像分析仪的精确性和准确性中的应用

多年的肿瘤病理和肿瘤临床研究已经证明，细胞核DNA倍体的异常变化对恶性肿瘤的诊断、疗效估价、预后评估具有重要的指导意义。图像分析仪已广泛应用于测量和分析肿瘤细胞核DNA倍体。日前，我国图像分析仪的生产厂家众多，但至今没有一个统一的国家或学会的技术标准，不可避免地导致国产图像分析仪在质量和性能上存在差异。     

胃癌细胞p16蛋白表达与DNA定量分析的相关性研究

目的探讨胃癌细胞p16蛋白表达情况及其与DNA含量的相关性及意义.方法应用流式细胞仪(FCM)和图像分析仪(ICM)检测41例胃癌细胞p16蛋白表达量和DNA含量,10例正常胃组织也用FCM测p16蛋白表达量.结果41例胃癌p16蛋白表达量明显低于正常胃组织(荧光指数分别为0.81±0.09和1.00±0.11,P<0.01),p16蛋白表达量与胃癌病理分级有关,随病理分级增高而降低(P<0.05),与胃癌患者的性别、年龄无关.胃癌DNA相对倍体均值(U值)范围为1.14～2.98,U值随病理分级增高而升高(P<0.01).同时,胃癌p16蛋白表达量与DNA含量呈负的直线相关(P<0.01).结论p16蛋白的异常表达与胃癌的发生发展、恶性程度有关,并具有较重要临床意义.p16蛋白的表达水平可作为临床判断预后的指标之一.     

图像分析仪在测定肿瘤细胞DNA含量,倍体中的若干问题

图像分析仪是目前用于肿瘤细胞ＤＮＡ含量与倍体检测的两种主要仪器之一，将其应用于有关的科学研究和辅助临床诊疗涉及有机的３个阶段：（１）器官、组织、细胞标本的制备；（２）图像分析仪的测量和分析；（３）数据结果的应用。第１、３阶段与我们生物医学工作者密切相关，对此，在本文中提出可能存在的部分问题，提请使用者注意。第２阶段的关键是生产厂家提供具有优异工程技术性能、准确应用相关生物医学概念、符合肿瘤细胞ＤＮＡ倍体分析国际标准的算法规则和分析参数的图像分析仪，其相关问题将另文阐述。一、肿瘤细胞的ＤＮＡ含量、…     

肿瘤细胞核DNA含量测定与倍体分析的现状及发展趋势

国内、外大量病理和临床研究资料已经证明，肿瘤细胞DNA非整倍体是恶性肿瘤的特征性标志之一。测量和分析细胞核DNA含量与倍体对恶性肿瘤的病理诊断、恶性程度判定、疗效估价、预测预后具有重要价值’‘－‘’。目前，主要采用流式细胞术（flowcytometry，FCM）和细胞图像光度术（imagecytometryorcytoph。tometry，ICM）测量和分析细胞学样品单个完整肿瘤细胞的DNA含量和倍体。FCM检测的细胞数多，速度快，灵敏度和精度高，采用数学模型分析其群体的各细胞周期比率或倍体分布，使各样本的检测结果之间具有比较高的可比性，是目前肿…     

组织原位肿瘤细胞DNA含量与倍体分析的某些方法学问题

将图像分析仪用于肿瘤细胞ＤＮＡ含量与倍体的分析 ,其结果在辅助临床病理的诊断、治疗和预后评估方面确实能起到一定的参考作用。近几年 ,国内不少医院病理科购买了图像分析仪 ,但在使用中却常常忽视了非常重要的方法学问题 ;也由于该技术本身固有的缺陷尚未被突破以及未采用标准化的检测方法和指标 ,造成该技术未能正式应用于辅助肿瘤的临床诊断、治疗和预后评估 ,这应引起我们的高度重视。在组织切片上 ,原位测量和分析肿瘤细胞核的ＤＮＡ含量与倍体具有许多细胞学涂片不可比拟的优越性。然而 ,由于测量和分析方法本身处于不断完善之中 ,…     

胃癌的细胞增殖、DNA增殖分数和倍性与患者生存的关系

目的：探讨胃癌细胞增殖、DNA增殖分数(S+G2/M期)和倍性与胃癌患者生存和生物学特性的关系。 方法： 应用流式细胞术，分别分析胃癌细胞群体的增殖、DNA增殖分数和倍性。 结果： 胃癌细胞群体的增殖和增殖分数之间无明显相关关系(P>0.05)；它们与胃癌患者的生存亦无明显相关关系(P>0.05)。胃癌细胞群体的增殖和DNA倍性与胃癌临床病理参数之间无明显相关关系(P>0.05)。胃癌患者DNA二倍体组和异倍体组Kaplan-Meier生存曲线经Log-rank检验有统计学差异（P<0.05）。 结论： 胃癌细胞的增殖和增殖分数与胃癌患者的生存无明显关系； DNA倍性分析对胃癌预后的判断具有重要参考价值。     

儿童肾透明细胞癌的病理及其癌细胞DNA定量分析

目的：探讨儿童肾透明细胞癌病理形态特点及癌细胞DNA含量和倍体的关系。方法：对4例儿童肾透明细胞癌组织进行病理形态观察,并用图象分析仪对癌细胞DNA进行定量分析。结果：4例癌细胞胞质透明呈透明细胞且有乳头状结构,其中2例乳头状结构超过50％,3例均见钙化小体及明显出血坏死,DNA检测,DNA平均指数1．31,呈2倍体型,高2倍体型或亚4倍体型,结论：儿童肾透明细胞癌病理形态特点为癌细胞胞质透明,以乳头状结构为主,可见钙化小体,常伴出血,坏死,癌周肾小球,肾小管基本正常为特征,癌细胞DNA呈2倍体或高2倍体型或亚4倍体型。     

应用流式细胞仪结合阴道镜检测宫颈组织细胞异倍体的临床意义

本文探讨宫颈组织细胞DNA倍体分析系统在早期宫颈癌诊断中的价值。应用流式细胞仪检测了60例在阴道镜下作宫颈活体组织细胞DNA倍体分析异常的妇女,同期做病理检查并结合临床资料进行分析,发现DNA倍体分析系统作为早期宫颈癌筛查新方法,与阴道镜联合应用可提高诊断宫颈癌前病变一宫颈上皮内瘤样病变（CIN）和早期宫颈癌的敏感性、特异性。     

胃癌DNA倍体分析及其TIMP-2和E-cadherin在胃癌中的表达

目的：研究胃癌中DNA倍体及其TIMP-2和E-cadherin的表达，探索胃癌侵袭转移的分子基础和可能机制。 方法:采用免疫组化技术检测E-cadherin、TIMP-2在99例胃癌，16例癌周正常黏膜，16例胃癌远处转移和25例胃癌转移阳性的淋巴结中表达情况；选取其中47例胃癌，6例癌周正常黏膜及4例胃癌远处转移标本采用流式细胞术检测DNA倍体及S期分数。 结果:TIMP-2表达与Borrmann’s 分型、淋巴结转移和浸润深度有关；E-cadherin表达与肿瘤细胞分化、Lauren’s 分型、Borrmann’s 分型、淋巴结转移和浸润深度有关。DNA异倍体率与分化和淋巴结转移有关，S期分数（SPF）与肿瘤大小、分化及淋巴结转移有关。而且在癌与癌周非癌黏膜之间E-cadherin表达、DNA异倍体率和S期分数的差别具有统计学意义；TIMP-2与 E-cadherin之间无相关性；E-cadherin表达与DNA倍体及S期分数呈正相关。 结论:随着肿瘤的演进和异质化，TIMP-2和E-cadherin 的异常表达及DNA异倍体和高S期分数也相应逐渐增加，提示它们在胃癌演进过程中起着关键作用，可以作为胃癌生物学行为的客观标志物。而且，这几种因素间的相互作用更加速了肿瘤演进过程。     

图像分析系统在检测生后发育期小鼠肝细胞核多倍体化中的应用

目的　探讨发育期小鼠肝细胞核DNA倍体动态变化规律。方法　本文选取不同发育时期健康雄性小白鼠的肝细胞涂片 ,Feulgen染色 ,TIGER细胞图像分析仪测量单个完整肝细胞核的DNA含量与倍体 ,分析发育时期小鼠不同DNA倍体肝细胞核所占的比率。结果　不同鼠龄小鼠同一倍体的肝细胞核DNA含量大致相同 ;二、四、八、十六倍体的肝细胞核间的比值均接近 2、4、8,它们的CV <9% ;鼠龄 2 - 2 0天的小鼠以二倍体肝细胞核为主 ;35天小鼠具有四种不同DNA倍体的肝细胞核 ,各DNA倍体肝细胞核所占比例依次为 :4C >8C >2C >1 6C ;6 0天小鼠偶见三十二倍体肝细胞核。结论　小鼠肝细胞多倍体化出现的时间比已有实验结果更早、程度更强     

用流式细胞术检测胃癌p53蛋白的表达

目的：检测并比较胃癌及癌前病变p53蛋白表达水平,探讨p53蛋白在胃癌发病中的意义。方法;采用间接免疫荧光标记,流式细胞术分析。以DNA指数、增殖指数、荧光指数为分析指标。结果：胃癌、不典型增生及肠上皮化生的荧光指数（FI）值与正常胃粘膜相比差异均有显著性（P＜0．05）,前三者与浅表性胃炎相比差异也都有显著性（P＜0．05）。不同病变间的比较,可见胃癌FI高于不典型增生及肠上皮化生（P＜0．05）。不典型增生p53蛋白阳性率为27％,胃癌为68％,在不典型增生及胃癌病例中,其异倍体的FI值、增殖指数（PI）值和p53蛋白阳性率与二倍体者相比差异都有显著性（P＜0．05）。结论：胃癌组织p53蛋白的表达水平高于癌前病变及正常胃粘膜组织,随病变向恶性转化,p53蛋白、PI及异倍体率均增高。检测p53蛋白表达水平对胃癌的诊断具有一定意义。     

Clinical Application of Flow Cytometry for DNA Analysis of Solid Tumors

Recent developments of flow cytometry (FCM) technology which make multiple correlative biological measurements on normal and neoplastic cells is affecting areas of diagnostic pathology as well as research fields, and a general understanding of FCM techniques is essential for pathologists. Today, FCM DNA measurement of tumors also becomes routine in the clinical and/or pathological laboratory for aid in cancer diagnosis and cancer treatment. It can also contribute to diagnosis of tumors as a supplemental method to conventional histopathology, and DNA ploidy and the percentages of S-phase fraction are considered as complementary prognostic parameters independent of the stage of disease. This article reviews clinical applications of flow cytometry focusing on the DNA measurements of solid tumors, and related practical issues, such as the methodology for nuclear DNA measurement, interpretation of DNA histograms and the relationship of DNA ploidy and S phase fraction to clinical and pathological features of human solid tumors.     

DNA倍体分析及Ki-67检测对肾实质小肿瘤性质和意义的探讨

目的 探讨肾实质小肿瘤的性质及其生物学行为,为临床进一步治疗和预后判断提供病理学依据。方法 应用流式细胞术和免疫组织化学方法,对48个直径≤3cm的小肿瘤和39个直径＞3cm的大肾癌的DNA倍体形式及Ki-67增殖状态进行了比较研究。结果 48个小肿瘤中,6个腺瘤均为二倍体,Ki-67增殖指数（PR）均＜1％。42个透明、颗粒细胞型小肿瘤异倍体出现率为16.7%,与39例大肾癌的33.3%差异无统计学意义;42个小肿瘤部Ki-67 PR与分级相关。结论 小肿瘤中既有腺瘤也有腺癌,肿瘤大小不能作为区分它们的标准。DNA倍体及Ki-67 PR可作为肾实质肿瘤生物学行为和预后判断的重要指标。     

p53 and c-erbB-2 expression in schistosomal urinary bladder carcinomas and schistosomal cystitis with premalignant lesions

The purpose of this study was to ascertain whether DNA ploidy status and S-phase fraction affected the prognosis of patients with carcinoma of the thyroid gland. We reviewed all malignant thyroid tumours diagnosed in Iceland from 1955 to 1990. In all, 494 thyroid carcinomas were diagnosed during that period. By analysing tumour material from paraffin blocks by flow cytometry we were able to evaluate the ploidy status in 424 tumours and the S-phase value in 417 tumours. We detected aneuploid cell populations in 9.7% of papillary carcinomas, 24.3% of follicular carcinomas, 42.9% of medullary carcinomas and 78.6% of anaplastic carcinomas. Some 57% of tumours, mainly papillary carcinomas, had an S-phase value of less then 3%, whereas most of the other histological types of carcinoma, including all the anaplastic tumours, had an S-phase value of 3%. Univariate analysis indicated that both ploidy status and S-phase fraction were significant variables. When taking into account known prognostic variables of thyroid carcinoma in a multivariate analysis, however, neither ploidy status nor S-phase value proved significant. We conclude that DNA ploidy status and S-phase values are not independent prognostic factors in thyroid carcinoma.     

瘤体内DNA倍体异质性与食管癌组织学分级的关系

采用多点取材的方法，应用流式细胞术对８０例食管癌瘤体内ＤＮＡ倍体异质性进行研究。结果显示，倍体异质性肿瘤的检出率为４７．５％（３８／８０）；倍体异质性与组织学分级无关（Ｐ＞０．０５），而与患者的预后有关（Ｐ＜０．０５）。本研究提示，深入研究肿瘤ＤＮＡ倍体异质性将对判断患者预后，协助组织学分级具有重要的意义。     

Comparison of ploidy analysis by flow cytometry and image analysis in hydatidiform mole and non-molar abortion

Determination of DNA ploidy is useful in the diagnosis and classification of hydatidiform mole. Most reports of ploidy analysis in molar tissue have used DNA flow cytometry. Although image analysis cytometry offers theoretical advantages over flow cytometry, there have been few reports of ploidy analysis by image analysis in hydatidiform mole. We selected 47 cases and measured DNA ploidy by flow cytometry and image analysis cytometry in complete hydatidiform mole, partial hydatidiform mole and non-molar abortion. The two cytometry modalities were compared using kappa statistics. There was reasonable overall agreement between the two modalities (κ= 0.69) and when ploidy was stratified into diploid/polyploid and triploid categories there was near perfect agreement (κ= 0.93). Aneuploid cell populations, which were not evident on flow cytometry, were identified by image analysis in a significant proportion of complete and partial hydatidiform moles and in a small number of non-molar abortions. Flow cytometry and image analysis cytometry yield comparable ploidy information, useful in the diagnosis and classification of hydatidiform mole. Image analysis cytometry offers greater sensitivity in the detection of small non-diploid cell populations but the significance of this latter finding is uncertain.     

Quantitative DNA ploidy analysis of breast carcinoma: A study of the effects of joint photographer experts group (JPEG) compression on DNA ploidy images

Telepathology usage in the past has typically been a qualitative procedure rather than a quantitative measurement. DNA ploidy using image analysis has been favorably compared to DNA ploidy analysis by flow cytometry in numerous publications. A step from DNA ploidy analysis using conventional image analysis to DNA ploidy analysis using stored images allows DNA ploidy analysis by image cytometry to become a powerful tool in telepathology. Remote DNA ploidy analysis using stored images has an impact on the field of pathology, as not every hospital or laboratory can afford to perform this type of specialized testing. However, images have large data files and require lengthy transmission times over communication systems to other computers. Joint Photographer Experts Group (JPEG) compression is a computer algorithm that allows the file size of an image to be reduced in order to decrease transmission times to another computer. A study was initiated to investigate the effects of JPEG compression on images of Feulgen stained breast tumor touch preps and the resulting DNA ploidy histograms. Diagn Cytopathol 1996;15:231–236. © 1996 Wiley-Liss, Inc.     

Application of the crypt isolation technique to the flow cytometric analysis of DNA content in gastric carcinoma

An inevitable limitation of conventional flow cytometric analysis of gastric cancer DNA content is that the preparations of tumor cell nuclei are contaminated with stromal cell nuclei. Using the crypt isolation technique, we separated tumor tissues from stromal tissues and analyzed the DNA content in samples of pure gastric cancer cells (64 intestinal-type and 46 diffuse-type) by flow cytometry. Morphologically, crypts from well-differentiated and moderately differentiated adenocarcinomas usually showed large tube-like or sheet structures, whereas tumor tissues isolated from poorly differentiated adenocarcinomas usually exhibited small tumor cell clumps or clusters of varying sizes. Tumor ploidy was divided into diploid, aneuploid, and multiploid subgroups. Aneuploidy and multiploidy were observed in 12% (13 of 110) and 64% (71 of 110) of gastric cancers, respectively. A high frequency of DNA aneuploidy or multiploidy was associated with intestinal-type tumors, but not with any of the other clinicopathologic variables tested. In contrast, high S-phase fraction values demonstrated a close association with tumors with abnormal ploidy, advanced stage, intestinal type, and late TNM stage. Our results suggest that S-phase fraction may be a more useful indicator of aggressive behavior in gastric cancers than DNA aneuploidy. To our knowledge, the present study is the first to report flow cytometric DNA content in a large number of gastric cancer samples obtained using the crypt isolation technique.     

p57KIP2表达结合流式细胞术分析对葡萄胎的辅助诊断价值

目的 研究父源性印迹基因p57KIP2蛋白表达结合DNA倍体分析对葡萄胎的辅助诊断意义.方法 收集32例具有DNA多态性遗传学分析资料的胎盘绒毛水肿病例,以遗传学分析结果作为葡萄胎的诊断依据.全部病例进行常规组织学检查、p57KIP2免疫组织化学染色(EnVision法)及流式细胞术DNA倍体分析,采用双盲法与遗传学分析结果对照.结果 32例绒毛水肿病例的遗传学分析证明,21例为完全性葡萄胎(CHM),7例为部分性葡萄胎(PHM),4例为水肿性流产.CHM p57KIP2免疫组织化学表达的阴性率为95.2%(20/21).非CHM的p57KIP2免疫组织化学全部阳性(11/11).p57KIP2免疫组织化学与DNA多态性分析结果的吻合率为96.9%(31/32).流式细胞术分析发现,7例PHM均为三倍体核型;21例CHM中有14例为二倍体,7例为四倍体;4例水肿性流产均为二倍体或近二倍体.结论 p57KIP2免疫组织化学阴性是CHM的可靠标志物.p57KIP2免疫组织化学结合流式细胞倍体分析可明确区分三种胎盘绒毛水肿性病变.p57KIP2免疫组织化学阴性支持CHM,p57KIP2免疫组织化学阳性且为三倍体为PHM,p57KIP2阳性且为二倍体支持水肿性流产.