胃癌细胞DNA含量、AgNOR计数与其生物学特性关系的研究

本文使用图像分析技术测定了正常胃粘膜、萎缩性胃炎、不典型增生及胃癌共８９例细胞ＤＮＡ相对含量及ＤＮＡ倍体分布;并用银染方法对８９例细胞核仁形成区嗜银蛋白进行定量分析。结果显示：不典型增生往往可观察到与胃癌相似的ＤＮＡ核型即高异倍体的出现,高异倍体的出现可能是重要的癌前标志;在胃粘膜病变中,ＡｇＮＯＲ值随病变异型程度的加重而递增,各组间差异均有非常显著性意义（Ｐ＜０．０１）。早期癌、浸润癌和转移癌间,以及癌转移过程中粘膜层、肌层、淋巴结内癌细胞ＤＮＡ含量无明显差异;ＡｇＮＯＲｓ计数均值差异亦无非常显著性意义（Ｐ＞０．０５）。随癌细胞分化程度升高,ＤＮＡ含量增多及异倍体出现率升高;各期胃癌除粘激腺癌与分化型腺癌差异无显著性意义外,余各型胃癌ＡｇＮＯＲｓ值随分化程度升高而增多,差异有显著性意义（Ｐ＜０．０５或Ｐ＜０．０１）。因此,ＤＮＡ含量及倍体分析,ＡｇＮＯＲｓ计数与胃癌的生物学行为密切相关,可作为胃癌早期诊断,癌前病变预测以及组织分级的一种重要指标。     

AgNORs异型颗粒在乳腺癌细胞核中的观察分析

本文报道了核仁组成区嗜银蛋白（ＡｇＮＯＲｓ）在乳腺癌细胞核中的一组异型颗粒——字母样结构“ＣＥＩＪＲＳＴＵＲＹＸ”等,乳腺良性病变无此结构。研究结果表明,字母样结构在浸润性导管癌中多见,在乳头状癌中未见。结合乳腺癌的临床分期分析,临床分期越高,有字母样结构的乳腺癌所占比分也越高。同时,有字母样结构的乳腺癌组其ＡｇＮＯＲｓ含量和颗粒形态的异型性均明显高于无字母样结构的乳腺癌组。在讨论中认为,ＡｇＮＯＲｓ字母样结构对鉴别诊断乳腺癌可能有一定的形态学参考价值     

乳腺癌AgNOR计数与癌胚抗原的免疫组化研究

应用核仁组成区嗜银蛋白（ＡｇＮＯＲ）技术及癌胚抗原（ＣＥＡ）免疫组化染色对９８例乳腺良、恶性病变进行对比研究。结果表明：ＡｇＮＯＲ计数与肿瘤增殖活跃程度及生物学行为是一致的。乳腺癌中ＡｇＮＯＲ计数显著高于良性病变（Ｐ＜０．０１）。浸润癌ＡｇＮＯＲ计数比其他类型乳腺癌高。ＣＥＡ染色在乳腺良性病变中基本阴性,恶性病变中阳性率８０．８％。乳腺癌中ＡｇＮＯＲ计数与ＣＥＡ分布之间呈线性相关（ｒ＝０．８２,Ｐ＜０．０５）。ＣＥＡ阳性乳腺癌组与ＣＥＡ阴性组ＡｇＮＯＲ计数差异显著（Ｐ＜０．０５）。提示：ＡｇＮＯＲ定量研究和ＣＥＡ分布在乳腺良、恶性病变的鉴别及肿瘤恶性程度的研究中具有相似的参考价值。     

甲状腺良、恶性病变中增殖细胞核抗原、P_(53)蛋白与AgNOR的检测及相关性研究

本文应用ＬＳＡＢ免疫组织化学方法及ＡｇＮＯＲ技术对７３例甲状腺良、恶性病变ＰＣ－ＮＡ、Ｐ５３蛋白及ＡｇＮＯＲ进行了检测并对其相关性进行了研究。结果显示①ＰＣＮＡ表达在良、恶性病变中有显著差异（Ｐ＜０．０１）;②ＡｇＮＯＲ颗粒均数及其形态在良、恶性病变中有较显著差异;③ＰＣＮＡ表达与ＡｇＮＯＲ计数呈显著正相关性（Ｐ＜０．０１）;④Ｐ５３蛋白在良、恶性病变中的表达均呈阴性,只是在淋巴结转移灶内呈阳性表达。根据研究我们认为检测甲状腺良、恶性病变中ＰＣＮＡ及ＡｇＮＯＲ颗粒对甲状腺良、恶性病变的鉴别、预后及对临床治疗均有意义     

肿瘤核仁形成区形态定量诊断的误差及规范化分析

本文采用包括自动图象分析技术在内的ＡｇＮＯＲ形态定量学方法,以大肠肿瘤为模型,进行了ＡｇＮＯＲ定量形态学研究的误差分析,以探讨肿瘤ＡｇＮＯＲ定量诊断规范化的可能。结果表明,染色条件、视场目标选择和细胞计数量是引起ＡｇＮＯＲ定量诊断的主要误差;恒定染色环境,正确选择欲测细胞及测定足够量的细胞是使ＡｇＮＯＲ形态定量诊断规范化的途径。     

正常造血细胞的核仁组成区计数定量研究

本文应用银染技术对正常造血细胞的核仁组成区进行了计数定量研究。结果显示,在粒系和红系中,随细胞的成熟,细胞中簇状ＡｇＮＯＲ的数量逐渐减少．而点状ＡｇＮＯＲ数量逐渐增多,无分裂能力的成熟细胞仅有少数点状ＡｇＮＯＲ。淋巴细胞中为一完全集结的银染颗粒,而巨核细胞内为各自分离的点状银染颗粒。本结果为正常造血细胞的核仁组成区提供了基础数值。     

介绍AgNORs的退染及复染技术

介绍ＡｇＮＯＲｓ的退染及复染技术朱建伟（山东省中医药研究所病理科济南,２５００１４）ＡｇＮＯＲｓ嗜银染色是临床及实验病理学特别是肿瘤病理学方面常用的染色方法。但有时因种种原因造成染色效果不理想,影响观察,如背景着色过深以及银染液沉渣等,需要进行重染。...     

AgNORs染色在病原菌检测中的特殊应用

ＡｇＮＯＲｓ染色在病原菌检测中的特殊应用胥维勇,杨红,李科,杨群（四川省人民医院病理科．成都６１００７２）核仁组成区嗜银蛋白ＡｇＮＯＲｓ染色法是近年来广泛采用的银浸润染色技术,它在判断细胞增殖状态,区别良恶性肿瘤研究时有一定意义［１］。我们应用ＡｇＮ...     

核仁组成区相关嗜银蛋白染色革新法

核仁组成区相关嗜银蛋白（ＡｇＮＯＲ）银染技术已广泛用以研究细胞生长活性,根据ＡｇＮＯＲ数目多少来判定肿瘤的良恶性和对肿瘤进行鉴别诊断。然而ＡｇＮＯＲ技术至今还存在背景非特异性银颗粒沉积问题而影响结果判定。本研究发现影响背景染色结果的主要因素是明胶的质量,采用Ｆａｒｍｅｒ’ｓ液可以清除背景染色,运用微波炉染色不仅可以缩短染色时间而且可以减少银用量。     

中国对虾cDNA文库的构建

中国对虾ｃＤＮＡ文库的构建ＣＯＮＳＴＲＵＣＴＩＯＮＯＦｃＤＮＡＬＩＢＲＡＲＹＯＦＳＨＲＩＭＰＰＥＮＡＥＵＳＣＨＩＮＥＮＳＩＳ（ＣＲＵＳＴＡＣＥＡ,ＤＥＣＡＰＯＤＡ）关键词中国对虾ｃＤＮＡ文库构建ＫｅｙｗｏｒｄｓＰｅｎａｅｕｓｃｈｉｎｅｎｓｉｓ,ｃＤＮ...     

Heterogeneity of DNA ploidy in gastric cancer.

Heterogeneity of DNA content was analyzed in 389 samples from 65 resected gastric cancers. Analysis of the samples revealed that there were 14 homogeneously diploid tumours. Six tumours were uniformly DNA aneuploid, each tissue block containing the same DNA index. The other 45 tumours (69%) varied in DNA content heterogeneity. In 39 of 45 tumours, there was a mixture of diploid and aneuploid samples, and 25 of the 39 tumours had a single aneuploid stemline. In 14 out of 39 tumours, there was also a mixture of diploid and aneuploid samples having two or more DNA aneuploid stemlines. In the remaining six tumours, different DNA aneuploid stemlines were contained in different samples without evidence of diploidy. When four or fewer samples were analyzed, only 50% of the tumours were diagnosed as having DNA content heterogeneity. On the other hand, 78% of the tumours showed DNA heterogeneity when 5 or more samples were analyzed. If the tumours had not been widely sampled, about a quarter of the tumours would have been mislabeled as diploid. The patients with tumours showing homogeneous diploidy survived longer than those with tumours showing a mixture of diploid and aneuploid stemlines. The survival rate was lowest for the patients with tumours having a mixture of diploid and multiple aneuploid stemlines, compared with those showing homogeneous diploid or a mixture of diploid and single aneuploid stemlines. The data from the current study clearly demonstrate the importance of adequate sampling in assessing the ploidy status of gastric cancers to identify groups of patients running different clinical course and prognosis.     

Dry mass, DNA and non-histone protein determinations in lung cancer cells

Summary Cell nuclei from two biopsies of bronchial mucosa, seven squamous-cell carcinomas and six small-cell carcinomas of the lung were investigated. DNA and non-histone protein (NHP) content were simultaneously determined in Feulgen—Naphthol Yellow S-stained smears by means of multiple plug cytophotometry. In addition, the nuclear dry mass of cells originating from the same populations was measured by interference microscopy.DNA histograms of carcinomas were characterized by DNA stemlines being situated in the diploid range in four out of six small-cell carcinomas and in the hyperdiploid to hypertetraploid range in six out of seven squamous-cell carcinomas.Polyploid values (up to 8 c) were seldom found in small-cell carcinomas whereas squamous-cell carcinomas showed a broader dispersion approaching the 16 c level.The similarity of the NHP distributions with the DNA histograms was more marked in small-cell carcinomas than in the squamous-cell carcinomas. In comparison with the NHP distributions of normal bronchial epithelial cells, those of carcinomas were shifted to higher values. The increased NHP content was found to be a more prominent sign of malignancy in small-cell carcinomas than the DNA mass. The increase in nuclear dry mass in carcinomas was mainly caused by the accumulation of NHP in tumour cell nuclei.     

Flow cytometric analysis of DNA aneuploidy in primary and metastatic human solid tumors

DNA histograms were measured by flow cytometry for 656 human solid tumors (365 primary and 291 metastatic). The proportion of aneuploid cells in cell suspensions obtained by mechanical disaggregation was significantly higher than those obtained after enzymatic disaggregation (collagenase + DNAse) of the same tumor. A strong correlation was observed between the values of DNA-indices measured after staining with propidium iodide and with 4',-6-diamidino-2-phenylindole (r = 0.97). Aneuploid cells were observed in 430 tumors (66%); 30 of these had two aneuploid stemlines, and two had three aneuploid stemlines. The overall frequency of aneuploidy was 61% among primary and 71% among metastatic tumors. The median value of the DNA index was 1.67 for 224 primary aneuploid tumors and 1.68 for 206 metastatic aneuploid tumors. For most diseases, the largest proportion of aneuploid primary and metastatic tumors had DNA-indices in the hypertriploid region. No major differences in frequency and degree of aneuploidy was observed between primary and metastatic tumors. For carcinomas of the bladder and prostate, frequency of aneuploidy was higher among poorly differentiated, than among moderately and well-differentiated tumors. For carcinomas of the breast and for sarcomas, tumors with DNA-indices of greater than 2.0 were observed mostly in the poorly differentiated group. For patients with carcinomas of the bladder and prostate most tumors at earlier stages of disease were diploid; whereas most tumors at later stages of disease were aneuploid. For patients with carcinomas of the ovary, colon, and kidney, no relationship between stage of disease and aneuploidy was evident.     

Molecular characterization of Entamoeba histolytica RNase III and AGO2, two RNA interference hallmark proteins

Entamoeba histolytica, a protozoan parasite with variable DNA content and complex ploidity, has defied most efforts aimed at gene depletion using classical genetic methods. In this study, we identified and characterized two proteins involved in the RNA interference (RNAi) pathway, RNase III and AGO2. Our results strengthen the findings that an RNAi pathway does exist in this parasite.     

Modal DNA values of normal and malignant urothelial cells of the bladder in relation to nuclear size

In a study of the correlation between mean nuclear size and DNA content in urinary bladder carcinoma, the modal DNA values of cell suspensions from 125 biopsies, obtained from 86 patients with malignant or normal urinary bladder epithelium, were analyzed by flow cytometry (FCM). Light microscopic measurements of nuclear size were carried out on smears from the same material. The results were correlated to the histopathologic stage and grade. The mean nuclear volumes were significantly larger in diploid tumor cells than in cells of normal epithelium. Aneuploid tumors showed significantly larger nuclei than did diploid tumors. Although there was a significant correlation between increases in the nuclear volume and in the DNA content, there was some overlapping between various grades of malignancy: mean nuclear volumes in aneuploid grade 2 tumors did not differ from those in aneuploid grade 3 tumors. A combination of FCM and morphometry discriminated all but 16% of the tumors from the normal cases. It is concluded that FCM and morphometry are complementary and can be used for the objective characterization of urinary bladder carcinomas.     

Comparison between slide and flow cytophotometric DNA measurements in breast tumors

Nuclear DNA analysis was performed in 37 human mammary adenocarcinomas in order to elucidate the difficulties and pitfalls connected with the interpretation of DNA histograms obtained using different methodologic approaches. For each tumor, DNA profiles were obtained by means of slide microspectrophotometry on a fine needle aspirate, slide cytophotometry on a 4-micron histologic section and flow cytometry on a suspension prepared from a cube of fresh tissue. When the DNA histograms were interpreted according to criteria usually applied to discriminate low-grade malignant tumors from high-grade malignant tumors, some tumors classified as euploid by one method were classified as aneuploid by another method. The main reasons for this weak correlation seem to be in specimen preparation and in tumor cell representation within the specimen between the methods. Another reason is that slide and flow techniques exhibit different sensitivities for malignancy-associated nuclear DNA changes: minor alterations of the DNA content of the tumor stemlines seem to be more exactly reported by means of the flow technique whereas structural alterations of the nuclear chromatin seem to be more sensitively recorded by means of the slide technique. It is suggested that thorough control of each step of the various DNA analysis procedures and the use of information obtainable by slide and flow techniques taken together may significantly improve the prognostic value of DNA measurements.     

Comparative cytofluorometric DNA analysis of pleomorphic adenoma and adenoid cystic carcinoma of the salivary glands.

The nuclear DNA content in the tumor cells of 17 pleomorphic adenomas (PA) and 26 adenoid cystic carcinomas (ACC) was assayed by cytofluorometry to clarify the etiology of the differences in their biological nature. Aneuploidy was found in only two solid-pattern ACC samples; all the other samples had diploid stemlines. With respect to the polyploid cell rate and the S + G2M fraction, the differences between the four histological patterns of PA were not significant. In the ACC, the S + G2M fraction was significantly higher in solid pattern than in cribriform or trabecular pattern tumors, which may indicate that proliferative activity is higher in the solid pattern than in the other two. The polyploid cell rate in ACC was significantly higher in the solid pattern than in the cribriform pattern carcinomas. There was no significant difference in the mean polyploid cell rate between PAs and ACCs. The mean S + G2M fraction was significantly higher in all histological patterns of ACCs than in PAs. These results suggest that proliferative activity is greater in any one of the histological patterns of ACC than in PA. These findings may largely explain the biological differences between the two tumors.     

Cytophotomeric DNA-analysis of aspirated cells from prostatic carcinoma.

In the present study material obtained from prostatic lesions by transrectal aspiration biopsy was subjected to a comparative morphologic and cytophotometric DNA analysis. Based on the morphologic pattern, the clinical material was divided into benign lesions (prostatic hyperplasia), suspected prostatic malignancy and highly, moderately and poorly differentiated prostatic carcinoma. The cytochemical analysis, based on quantitative cytophotometric measurements of Feulgen stained nuclei, showed that cell nuclei from benign lesions (prostatic hyperplasia) exhibited the normal diploid amount of DNA. Contrary to this, cell populations from prostatic malignancies, were characterized by various degrees of heteroploidy, i.e. the existence of cells with nuclear DNA quantities increased above the normal diploid level. A general correlation between degree of heteroploidy (frequency of heteroploid cells) and degree of clinical malignancy seemed to exist; high grade malignant prostatic carcinoma (poorly differentiated) exhibited a pronounced degree of heteroploidy with more or less distinct aneuploid stemlines, whereas low-grade prostatic carcinomas (highly differentiated) were more similar to benign cell populations, in showing a large proportion of cells with a diploid DNA quantity suggesting the existence of diploid or near diploid stemlines. Cases morphologically classified as moderately differentiated prostatic carcinoma, which previously have been shown to exhibit individual variations in degree of clinical malignancy, also showed large individual variations in degree of hetyroploidy. Approximately half of these cases had cytochemical DNA characteristics similar to that of highly differentiated prostatic carcinoma in showing a modal DNA value in the diploid region, while the other half showed cytochemical characteristics similar to that of poorly differentiated prostatic carcinoma, i.e. aneuploid DNA distributions. However, no morphologiifferentiated prostatic carcinoma. In conclusion it can be stated that the present investigation suggests the possibility that quantitative cytochemical DNA analysis may be used in combination with, and offer additional information to, morphologic analysis in the malignancy grading of prostatic carcinoma. A future clinical follow-up, now in progress, will hopefully give a more definite answer to that idea.