Morphometrical Analysis of Nuclear Abnormality of Tubular Tumors of the Stomach with Image Processing

Nuclear size and shape on histological specimens were used to quantify nuclear abnormality in gastric tubular tumors. A valid technique for this morphometrical analysis by image processing was developed. Furthermore, the present study examined the nuclear size and shape to determine if they were reliable criteria to be used for differential diagnosis. Nuclear area, maximum diameter and maximum width on the histological specimens were defined as factors to represent nuclear size. Nuclear aspect ratio represented nuclear shape. Histological features revealed varied degrees of condensation of nuclei; this was due to different protocols for preparing the histological specimens. This finding indicated that the size of intermediate lymphocytes on the same specimens should be measured as a control in order to estimate the degree of nuclear condensation. The present study also demonstrated that at least 200 nuclei should be measured on the histological specimens to obtain accurate nuclear size and shape. Histologically, the nuclei in benign tubular adenoma and borderline lesions tended to be spindle-shaped, while those in well-differentiated type tubular adenocarcinoma tended to be round and swollen. The nuclei in borderline lesions were larger than those in benign tubular adenoma and smaller than those in well-differentiated type tubular adenocarcinoma. Our results for the above 4 factors corresponded well with these histological findings. The nuclear size and shape on histological specimens were concluded to be reliable criteria for assessing nuclear abnormality in gastric tubular tumors and useful ones for differential diagnosis.     

Morphometrical Analysis of Structural Abnormality of Tubular Tumors of the Stomach with Image Processing

Four indices, index of tubular density, index of dispersion of tubular size, degree of complexity of tubular shape and frequency of complex-shaped tubuli, were defined to quantify the structural abnormalities of gastric tumors in morphometrical analysis by image processing. The values of each index corresponded well with the degree of each structural abnormality found pathologically. These indices were considered to be valid for representing their respective structural factors. There were significant differences among the mean values of the scores calculated by a formula using all 4 indices of benign tubular adenoma, borderline lesion and well-differentiated type tubular adenocarcinoma (P<0.05). Therefore, the discriminant formula was considered to be valid for integrating these indices and for representing structural abnormality of gastric tubular tumors.     

Morphometrical analysis of structural abnormality of tubular tumors of the stomach with image processing

Four indices, index of tubular density, index of dispersion of tubular size, degree of complexity of tubular shape and frequency of complex-shaped tubuli, were defined to quantify the structural abnormalities of gastric tumors in morphometrical analysis by image processing. The values of each index corresponded well with the degree of each structural abnormality found pathologically. These indices were considered to be valid for representing their respective structural factors. There were significant differences among the mean values of the scores calculated by a formula using all 4 indices of benign tubular adenoma, borderline lesion and well-differentiated type tubular adenocarcinoma (P less than 0.05). Therefore, the discriminant formula was considered to be valid for integrating these indices and for representing structural abnormality of gastric tubular tumors.     

Objective quantification of grade of atypia in epithelial tumors of the stomach by image processing

An image processing technique to extract tubular and nuclear areas from histological specimens stained with hematoxylin was established. Two indices representing tubular density and nucleus-to-cytoplasm ratio of glands were examined in intestinal-type tumor of the stomach. In specimens clearly stained with hematoxylin, significant differences of brightness among nuclei, cytoplasm and stroma can be obtained. Image processing by computer enabled us to identify nuclei, cytoplasm and stroma by utilizing the differences of brightness among them. Both indices tended to increase with severity of histological atypia and their values in the tubular adenoma group were significantly different from those in the tubular adenocarcinoma group (P less than 0.05). Therefore, these two indices could be regarded as valid and objective measures of the grade of histological atypia. Eighty-four percent of tubular adenoma and tubular adenocarcinoma cases were classified correctly with the discriminant formula and critical value calculated from the two indices on the basis of measurement of the images magnified by 40 times. There is, however, still a non-negligible overlap of discriminant scores between the tubular adenoma group and tubular adenocarcinoma group. More indices representing histological atypia will be required to allow objective differential diagnosis between malignant and benign lesions.     

Objective Quantification of Grade of Atypia in Epithelial Tumors of the Stomach by Image Processing

An image processing technique to extract tubular and nuclear areas from histological specimens stained with hematoxylin was established. Two indices representing tubular density and nucleus-to-cytoplasm ratio of glands were examined in intestinal-type tumor of the stomach. In specimens clearly stained with hematoxylin, significant differences of brightness among nuclei, cytoplasm and stroma can be obtained. Image processing by computer enabled us to identify nuclei, cytoplasm and stroma by utilizing the differences of brightness among them. Both indices tended to increase with severity of histological atypia and their values in the tubular adenoma group were significantly different from those in the tubular adenocarcinoma group (P< 0.05). Therefore, these two indices could be regarded as valid and objective measures of the grade of histological atypia. Eighty-four percent of tubular adenoma and tubular adenocarcinoma cases were classified correctly with the discriminant formula and critical value calculated from the two indices on the basis of measurement of the images magnified by 40 times. There is, however, still a non-negligible overlap of discriminant scores between the tubular adenoma group and tubular adenocarcinoma group. More indices representing histological atypia will be required to allow objective differential diagnosis between malignant and benign lesions.     

Well-differentiated adenocarcinoma of the papilla of vater--report of a case

Reported is the case of a 64-year-old female who was admitted to our clinic because of hyperamylasemia. Duodenal endoscopic examination revealed a tumor at the papilla of Vater. A biopsy specimens taken from the top of the tumor showed a well-differentiated adenocarcinoma. Under the diagnosis of cancer of the papilla of Vater, pancreaticoduodenectomy was performed. The first histological diagnosis was carcinoma in adenoma, but from the results of nucleus-gland ratio examined from 6 parts of the "adenoma", all of which were over 0.50, we finally concluded this case as de nove well-differentiated adenocarcinoma, and not carcinoma in adenoma. Thus, there is a possibility that past reports of adenoma or carcinoma in adenoma of the papilla of Vater might contain de novo well-differentiated adenocarcinoma.     

大肠隆起性病变核形态计算机鉴别诊断的应用研究

应用自动图像分析的方法，对内镜下检出的大肠隆起性病变１８３例活检标本的细胞核形态特征进行定量研究。结果显示非腺瘤性息肉、腺瘤及腺癌细胞核形态的各项参数值均随病变恶性程度的增加而增大，参数值在正常及各项隆起性病变组间的差异具显著性（Ｐ＜０．０１）。将上述数据输入计算机进行多因素逐类判别并建立函数判别式，通过回代矩阵分析与原诊断的符合率为８９％，其中大肠癌、腺瘤、非腺瘤性息肉分别为８６％、８５％、９５％，显示出一定的鉴别诊断功能和临床应用价值。     

Clinical usefulness of microsatellite instability for the prediction of gastric adenoma or adenocarcinoma in patients with chronic gastritis

To assess a role of microsatellite instability (MSI) in the development of gastric adenocarcinoma or adenoma from chronic gastritis, we analysed mutations of five microsatellite loci in gastritis, adenoma and adenocarcinoma retrospectively. Gastric mucosa was biopsied from the same area in each patient at different periods and examined for MSI. Only one of 55 patients with chronic gastritis revealed MSI-H phenotype and the other 54 patients showed microsatellite stable (MSS) phenotypes. In six of 17 patients with gastric adenoma or well-differentiated adenocarcinoma, MSI-positive phenotypes were demonstrated. Interestingly, all of six patients showing MSI, including three high-level MSI (MSI-H) cases and three low-level (MSH-L) cases, had already revealed MSI at the stage of chronic gastritis. In two of three MSI-H cases, the identical MSI patterns had been observed at the stage of gastritis 1.5-7 years before the final diagnosis of adenocarcinoma. The adjacent gastritis mucosa within 10 mm from the carcinoma demonstrated MSI as well. MSI was not found in any of 35 patients with Helicobacter pylori infection, but found in one of 30 patients without infection. Moreover, two of three cases of gastric adenoma or well-differentiated adenocarcinoma with MSI-H at the stage of chronic gastritis showed no evidence of Helicobacter pylori infection throughout the observation periods. These results indicate that MSI in biopsy specimens at the stage of chronic gastritis may predict the risk of the progression to adenoma and well-differentiated adenocarcinoma, and that Helicobacter pylori infection itself may not induce MSI directly in the gastric mucosa.     

High incidence of nuclear accumulation of p53 protein in gastric cancer.

The accumulation of p53 protein in the nuclei of cancer cells is known to correlate well with the presence of mutations in the p53 gene. We therefore investigated the immunohistochemical reactivity of the anti-p53 antibody, PAb1801, in specimens taken from 149 cases of primary gastric cancer and processed by acetone fixation, in order to elucidate the incidence and clinicopathological significance of p53 alterations in gastric cancer. Thirty-four out of 99 (34%) advanced gastric cancers and 11 out of 50 (22%) early gastric cancers showed positive reactions in the nuclei. The nuclei of non-cancerous cells, including gastric glandular epithelial cells, however, were not stained. Histopathologically, a nuclear accumulation of p53 protein was seen frequently in papillary adenocarcinoma, well- to moderately-differentiated tubular adenocarcinoma and poorly-differentiated adenocarcinoma with solid nests or focal tubular structures (43/101, 43%), but was rarely seen in signet-ring cell carcinoma, mucinous adenocarcinoma or poorly-differentiated adenocarcinoma growing in a scattered manner (2/48, 4%). There was no correlation between stainability of p53 protein and clinicopathological features such as depth of tumor invasion, microscopic lymphatic invasion, microscopic venous invasion, nodal involvement and clinicopathological stage in papillary adenocarcinoma, well- to moderately-differentiated tubular adenocarcinoma and poorly-differentiated adenocarcinoma with solid nests or focal tubular structures. The results suggest papillary adenocarcinoma, well- to moderately-differentiated tubular adenocarcinoma and poorly-differentiated adenocarcinoma with solid nests or focal tubular structures to share a common carcinogenetic pathway in which mutation of the p53 gene has an important role to play at a relatively early stage. Additionally, we showed the applicability of immunohistochemical detection of p53 protein in endoscopic biopsy material routinely formalin-fixed. The current method may be of some help in routine practice in discriminating between normal, precancerous and cancer cells in the stomach.     

Cx43、Skp2在卵巢上皮性肿瘤组织中的表达及临床意义

背景与目的:间隙连接蛋白43(connexin43,Cx43)是间隙连接蛋白家族的主要成员,在多种肿瘤细胞中表达下降。它在许多细胞系以依赖间隙连接(Gapjunction)的途径发挥抑癌作用。最近研究发现它还可能通过抑制S期激酶相关蛋白2(S-phase kinase associated protein2,Skp2)的表达而起到抑制肿瘤生长的作用。Skp2是F鄄box蛋白家族的成员,它能特异性识别并促进某些调节G1期进程的关键性细胞周期调节物的降解,在许多肿瘤中表达升高。本研究检测Cx43和Skp2在卵巢上皮性肿瘤中的表达,探讨它们的表达与卵巢癌发展的关系,以及这两种蛋白表达的相互关系。方法:收集81例卵巢上皮性肿瘤的外科手术石蜡标本(良性13例、交界性12例、恶性56例),用免疫组化的方法检测Cx43和Skp2蛋白的表达水平。分析它们的表达水平与临床病理参数的关系以及二者的相互关系。结果:Cx43蛋白在卵巢上皮性良性、交界性及恶性肿瘤中阳性率分别为84.6%、66.7%和33.9%,经免疫组化半定量分析,其在上皮性卵巢癌中的表达水平明显低于良性肿瘤(P<0.01)和交界性肿瘤(P<0.01),在不同年龄及组织类型的卵巢癌中表达无显著性差异(P>0.05),而在中低度分化组、Ⅲ～Ⅳ期组及有淋巴结转移组分别明显低于高分化组(P<0.05)、Ⅰ～Ⅱ期组(P<0.05)及无淋巴结转移组(P<0.0     

Differential expression of peroxisome proliferator-activated receptor in histologically different human gastric cancer tissues

Gastric cancer cell lines express peroxisome proliferator-activated receptor gamma (PPARgamma), and treatment with PPARgamma ligands suppresses growth of subgroup of these cell lines. However, expression and subcellular distribution of PPARgamma in human gastric cancer tissues is still unknown. Therefore, expression and subcellular localization of PPARgamma were examined among different histological types of gastric cancer tissues. Immunohistochemical staining for PPARgamma was performed using biopsy specimens of human gastric cancer of various histological types, gastric adenomas, and intestinal metaplasia. All samples of intestinal metaplasia and most samples of gastric tumors, except for signet ring cell carcinoma, expressed PPARgamma in the epithelial cells. Most samples of signet ring cell cancer lacked PPARgamma expression. All samples of intestinal metaplasia expressed PPARgamma only in the cytosol. For adenoma, 90% was positive for PPARgamma in cytosol, and 40% was positive in nuclei, for well-differentiated adenocarcinoma, 80% was positive in cytosol, and 20% was positive in nuclei. For moderately differentiated adenocarcinomas, 70% was positive for cytosol, and 80% was positive for nuclei; for poorly differentiated adenocarcinoma, 30% was positive in cytosol, and 70% was positive in nuclei. The frequency of samples with positive cytosolic staining decreased as the differentiation stage turned from intestinal metaplasia to adenoma, well-, moderately-, and poorly-differentiated cancers. Simultaneously, there was a tendency toward an increased frequency of samples with positive nuclear PPARgamma staining as the differentiation stage transformed from intestinal metaplasia to poorly-differentiated cancer. There was a striking difference in subcellular localization according to the differentiation levels of gastric dysplastic cells. The findings also supported an intestinal metaplasia-adenoma-well-differentiated gastric cancer sequence, and signet ring cell cancer was suggested to be of a different lineage from other types of gastric cancers.     

Small-sized adenocarcinoma of the lung. Cytologic characteristics and clinical behavior

BACKGROUND: Although the cytologic characteristics of advanced adenocarcinomas of the lung have been described, the unique cytologic features of early adenocarcinomas have not been established. METHODS: Cytologic specimens from 193 patients with small-sized lung adenocarcinoma, up to 2 cm in dimension, were reviewed. Cytologic investigations were performed on sputum, bronchial brushings or washings, or fine-needle aspirates obtained from the patients preoperatively. The cytologic characteristics of early adenocarcinoma were confirmed with Image Processor for Analytical Pathology from Sumitomo Chemical Co., Ltd. RESULTS: Nuclear size, variations in nuclear size, appearance of nucleolus, and nuclear atypia of the adenocarcinoma cells were found to differ between the various histologic subtypes of adenocarcinoma. The nuclei in small but advanced adenocarcinoma were generally larger and showed more variation in size than those in early adenocarcinoma (localized bronchioloalveolar carcinoma). CONCLUSIONS: Surgically curable, early adenocarcinomas of the lung were found to have a unique cytologic appearance, including small nuclear size and slight variation in nuclear size. Using these unique characteristics, they can be distinguished easily from advanced adenocarcinomas.     

胃黏膜活检组织中腺瘤的临床病理及免疫表型分析

目的探讨胃黏膜活检组织中腺瘤的临床病理学特点、免疫表型、生物学行为。方法应用光学显微镜、免疫组织化学和随访的方法对52例胃黏膜活检病理诊断腺瘤的标本进行研究。结果胃腺瘤的病理组织学特点：核间变,表现为核的体积增大及形态不规则,并出现1或多个核仁,核分裂≤2/10HPF。腺体结构变化：腺瘤的腺管多比较一致,偶有腺体的大小及轮廓不规则,可伴有肠上皮化生。当核体积增大明显,核分裂＞2/10HPF,腺体出现不规则的扭曲、分枝状结构,提示腺瘤有发展。免疫表型：不同分子量角蛋白和肿瘤相关糖类抗原均有较高的阳性表达。CDX-2、APC、PTEN和CEA高表达与腺瘤的形成密切相关。黏蛋白Muc-2高表达与黏蛋白Muc-1低表达呈负相关,两种结合有助于判断胃腺瘤的进展情况。survivin、bcl-2和p53蛋白的阳性表达率低,三者结合对胃腺瘤的恶性转化的判别有重要意义。Ki-67阳性细胞数在20%~40%之间,可作为细胞增殖程度,评价预后的有用指标。37～45月随访结果发现,治愈42.3%（22/52）、仍有腺瘤样增生38.5%（20/52）、有发展显示异型增生15.4%（8/52）、发展为黏膜内癌者3.8%（2/52）。结论胃腺瘤出现核体积明显增大,核分裂＞2/10HPF,腺体不规则的扭曲、分枝状结构;免疫组织化学染色显示survivin、bcl-2、PTEN、p53蛋白阳性表达增强,细胞增殖指数Ki-67＞40%,提示有癌变倾向,应内镜下黏膜切除。     

Nuclear/Nucleolar morphometry and DNA image cytometry as a combined diagnostic tool in pathology of prostatic carcinoma

Paraffin tissue sections from 50 patients with prostate adenocarcinoma were used to study nuclear and nucleolar morphometric features by image analysis. The results were compared to DNA ploidy and Gleason grade. In the examined histological samples nuclear and nucleolar areas were positively interrelated. It was also noticed that the higher the percentage of nucleolated nuclei, the bigger the nuclear and nucleolar areas. The morphometric characteristics did not differ significantly among the four grades of the examined specimens. In well-differentiated carcinomas the DNA index was lower than in the rest at a statistically significant level. Hypodiploid carcinomas were found to possess significantly bigger nuclear areas than any other DNA index group. Morphonuclear evidence of anaplasia and DNA aneuploidy may be used as diagnostic tools in prostate cancer in addition to Gleason grade.     

Analysis of gastric biopsy specimens group III by a revised group classification

A revised group classification of histology in gastric biopsy specimens was recently proposed by the Japanese Research Society For Gastric Cancer. The specimens judged as Group III by the revised classification contained; adenoma that used to be judged as Group III by the former classification, lesions difficult to distinguish between adenoma and carcinoma, lesions difficult to distinguish between atypical regenerative epithelium and carcinoma, and so on. Pathologists are requested to describe whether the lesion is adenoma or not and to consult with clinicians about the lesion in order to reduce the possibility that a case of well-differentiated adenocarcinoma is misdiagnosed as Group III by the revised classification.     

Quantitative nuclear morphology in the diagnosis of ovarian tumors of low malignant potential (borderline)

Ovarian tumors of low malignant potential or of borderline malignancy are characterized histologically by the association of malignant type of epithelial proliferation with a noninvasive pattern of growth. Because epithelial proliferation relates to features of nuclear morphology and chromatin structure and because of the difficulties to distinguish nuclear atypism seen in borderline malignancy from that in frankly invasive tumors by usual microscopic study, the authors concentrated their studies exclusively on computerized analysis of cell nuclei images. Using specially developed methods of analysis the authors described geometrical, optical, and structural differences among the epithelial and stromal cell nuclei of benign, borderline, and malignant ovarian tumors. The structural differences concern the pattern of chromatin condensation and suggest that the cytokinetic properties of the borderline tumors are intermediate to those of benign and malignant. The results demonstrate that the quantitative evaluations provide objective and reproducible data useful in the differential diagnostic of the borderline malignancy.     

Nucleolar prominence as a diagnostic variable in prostatic carcinoma.

The diagnosis of well-differentiated adenocarcinoma of the prostate can be difficult on needle biopsy specimens. Nucleolar prominence has proven to be a useful diagnostic variable, but its objective evaluation has had limited study. To find nucleolar criteria that might differentiate benign from malignant conditions, we examined 41 open prostatectomy specimens, 25 of which were removed for well-differentiated adenocarcinoma and 16 of which were removed for benign prostatic hypertrophy (BPH). Four acini of carcinoma, prostatic intraepithelial neoplasia (PIN), atypical adenomatous hyperplasia (AAH), nodular hyperplasia, and normal tissue were examined. The total number of nuclei with nucleoli 3 microns or greater (N'), the fraction of nuclei with nucleoli 3 microns or greater (N'/N), and the average diameters of nucleoli 3 microns or greater (AVG) were recorded. Hyperplastic and normal areas, when compared with carcinoma, had significantly smaller N',N'/N, and AVG values (P less than 0.005). The N' and N'/N values were significantly higher in hyperplasia when compared with normal acini (P less than 0.005). In addition, N' and N'/N values in PIN were significantly greater than those in AAH (P less than 0.0001). In comparing prostates with and without carcinoma, N' and N'/N were significantly different for hyperplastic areas. In only two cancer areas and one PIN area was the N'/N ratio less than 0.31, which was the highest value for either hyperplastic or normal areas. Although AVG also were significantly different, they did not improve discrimination between the groups. We conclude that N'/N ratios are useful in diagnosing well-differentiated prostatic adenocarcinoma on small tissue samples.     

Expression of placental alkaline phosphatase in gastric and colorectal cancers. An immunohistochemical study using the prepared monoclonal antibody

The authors developed monoclonal antibodies (MoAb) against human placental alkaline phosphatase (PLAP). Four specific MoAb reacting only with PLAP and two nonspecific MoAb reacting equally with isozymes of alkaline phosphatase (hepatic, intestinal, and placental) were obtained. Immunohistochemical staining with the specific MoAb showed that the cell membrane and cytoplasm of cancer cells were stained in gastric and colorectal carcinoma. The incidence of PLAP positivity was 23% (25 of 107) of all gastric carcinomas. Among gastric carcinomas, the 42% (13 of 31) positivity of highly differentiated carcinoma (papillary adenocarcinoma and well-differentiated tubular adenocarcinoma) was a significantly higher rate than that found in poorly differentiated carcinoma (poorly differentiated adenocarcinoma and signet-ring cell carcinoma, five of 41, 12%). The incidence of PLAP positivity was 11% (four of 35) in colorectal carcinoma. In contrast, gastric adenoma, intestinal metaplasia, and noncancerous tissue adjacent to cancer did not show staining. These results indicated that expression of PLAP was apt to occur in more highly differentiated gastric carcinoma and was highly specific for carcinoma in the gastrointestinal tract, although its incidence was not high.     

Flow cytometric analysis of nuclear dna content in tissues of colon cancer using endoscopic biopsy specimens

Flow cytometric assay of nuclear DNA in endoscopic biopsy specimens was evaluated in colon cancer patients. When the cells were divided into diploid cells and aneuploid cells, aneuploidy was observed in 63% (58 of 92) of the colon cancer patients. However, no clear relation was observed between the frequency of aneuploidy and the invasive depth, size, or histological type of colon cancer. Noncancerous portions of the colon tissues including colon adenoma or normal mucosa were mostly (96%, 87 of 91) diploid. Nuclear DNA content could be analyzed in the fresh biopsy specimens of colon cancer tissues and such investigation might be possibly valuable for further biological characterization of colon cancer in the usual procedure of clinical diagnosis for colonic malignancy before surgical operation or other treatment.     

Intraepithelial mononuclear lymphocytes and progression of stomach and colon cancers

Levels of intraepithelial lymphocytes were assayed in 60 bioptic samples taken from the stomach and colon. The pathologies included non-specific inflammation, benign epithelial proliferation, adenoma with grave epithelial dysplasia and adenocarcinoma. Among the methods used were routine histological staining, direct immunostaining with FITC-conjugated monoclonal antibodies against the human antigen CD4 and antibody Fab-fragment as well as an original procedure for DNA identification. In the colon, adenocarcinoma was distinquishable a high level of intraepithelial CD4(+) lymphocytes while, in the stomach, this was true for tubular adenoma with grave epithelial dysplasia, too.