Journal of Pathology and Translational Medicine

Volume 27(6); December 1993

Original Articles

Application of Argyrophilic Nucleolar Organizer Regions(AgNORs) in the Diagnosi of Hepatocellular Carcinoma.: Cheol Hee Yun, Sang Sook Lee, Eun Sook Chang; Korean J Pathol. 1993;27(6):553-560.

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Abstract: Necleolar organizer regions(NORs) ARE LOOPS OF DNA which transcribe to ribosomal RNA by RNA polymerase I. Since NOR-associated proteins are argyrophilic, silver staining method has been used for demonstration of AgNORs. The numbers and/or configurations of NORs may reflect the activities of cells in hyperplastic and neoplastic conditions. To evaluated the applicability of AgNORs in the diagnosis of hepatocellular carcinoma, the author had performed silver staining on the routinely processed, formalin-fixed, paraffin-embedded sections of 14 cases of normal liver(control), 23 cirrhotic liver, and 21 hepatocellular carcinoma. The results are summarized as follows: 1) The mean number of AgNORs per nucleus(mAgNOR) of normal liver, cirrhotic liver and hepatocellular carcinoma was 1.45+/-0.07, 2.53+/-0.38 and 5.52+/-1.63, respectively. The difference of mAgNOR between normal and cirrhotic liver, and between cirrhotic liver and hepatocellular carcinoma was statistically significant, respectively(p<0.01). 2) The percentage of nuclei showing five or more AgNORs per nucleus(pAgNOR) was 0.07% in normal liver, 7.59% in cirrhotic liver, and 60.49% in hepatocellular carcinoma. 3) AgNORs in hepatocellular carcinoma were large, pleomorphic and irregularly clumped, in addition to increase of mAgNOR and high pAgNOR. In conclusion, the increase of mAgNOR, high pAgNOR and large, irregular AgNORs are regarded as an additional helpful finding for the histopathological diagnosis of hepatocellular carcinoma.

A Pathological Study of Phenol Induced Hepatic and Gastrointestinal Lesions.: Dae Young Kang; Korean J Pathol. 1993;27(6):561-572.

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Abstract PDF: In an attempt to elucidate the pathological effects of phenol, the present study was undertaken in male Sprague-Dawley rats. The control group of animals was fed a basal diet, and potable underground water. The experimental group of animals was fed a basal diet and potable underground water containing 30ppm, 60ppm, and 1% phenol with once a week administration of dimethylnitrosamine(DMN) 10 mg/kg I.P. Each group of animals was sacrificed on the 3rd, 6th, and 9th month. The liver and gastrointestinal tract were examined light microscopically, along with transmission electron microscopic studies of the liver and scanning electron microscopic studies of the gastric mucosa. The results were as follows: 1) In the acute phenol intoxicated group, the liver showed fatty changes in the hepatocytes with mitochondrial membrane destruction and myelin figure formation. 2) In the chronic phenol intoxicated group, fatty changes in the liver were observed. In addition, there was chronic inflammation in the gastrointestinal tract, with gastric mucosal erosion and central necrosis of the hepatic lobules, especially in the high phenol contaminated water treated group. 3) As a result of the examination under the light microscope, the DMN treated group showed hyperplastic nodules and liver cell dysplasia, the degree of which was proportional to the duration of the experiment, and was more severe in the DMN + phenol treated group. 4) As a result of the examination under the electron microscope, fatty changes in the liver, pleomorphism of the mitochondria and loss or shortness of bile canalicular microvilli in the DMN + phenol treated group were more severe than in the group treated only with DMN. In summary, the results obtained by the present study indicate chronic highly concentrated phenol intoxication induce liver cell necrosis and chronic inflammatory with a hepatotoxin such as DMN.

Detection of Chromosome Aberration in Interphase Nuclei of Tumor Cells by Nonradioactive In Situ Hybridization Using Chromosome-specific Probes.: Sang Sook Lee; Korean J Pathol. 1993;27(6):573-580.

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Abstract PDF: An improved protocol for in situ hybridization(ISH) to routinely processed, paraffin-embedded tissue sections from lung carcinoma is presented. For this study, DNA probes for alpha-satellite chromosome 7 and 17 were used. The protocol to detect numerical chromosome aberrations involved treatment of sections with 1 M sodium thiocyanate prior to pepsin digestion, resulting in reproducible ISH reactions. The effect of avidin-biotin detection system. Four layer avidin methods and triple biotin methods, using avidin-PO, goat antiavidin, biotinylated antigoat IgG, avidin-PO or anti-biotin, biotinylated antirabbit IgG, avidin-PO, markedly enhanced the intensity of positive signals. More than 80% of the tumor and stromal cells showed distinct chromosome hybridization signals in 6 micrometer-thick sections. Lung carcinoma cells showed multiple chromosome signals(2~5 spots), contrasted by one or two signals in the stromal cells in the same section. These results suggest that chromosome polysomy can be reliably detected in tissue sections using in situ hybridization. This capability will prove to be an important tool for determining the underlying genetic basis for tumor development, tissue phenotype heterogeneity and progression by allowing genetic determination to be made on paraffin-embedded tissue sections where tumor histologic architecture is preserved.

Flow Cytometric DNA Analysis of Hepatocellular Carcinoma.: Young Lyun Oh, Yong Il Kim; Korean J Pathol. 1993;27(6):581-589.

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Abstract PDF: A flow cytometric analysis of the nuclear DNA content of solid tumors using paraffin-embedded tissues has become available since 1983, and its ploidy pattern has been designated as an important prognostic parameter in many human tumors. Hepatocellular carcinoma(HCC) is one of the most common malignant tumors among Koreans, but little information is consolidated about the significance of ploidy pattern. We measured the nuclear DNA content of 62 surgically resected HCCs and 45 non-neoplastic tissues from the surrounding parenchyma by flow cytometry. Aneuploid was detected in 18 cases(29.0%) in HCCs and 2 cases(4.4%) in nonneoplastic hepatic parenchyma(p<0.005). Correlations between the DNA ploidy pattern and various clinicopathologic findings of HCCs were analized. The mean tumor size was significantly different(p<0.05) between the aneuploid group(8.8 cm) and the diploid group(6.1 cm). Mean age of the aneuploid group was younger(47 year) than the diploid group(51 years), but the difference was not statistically significant(p=0.052). The DNA pattern did not show any meaningful correlation with the gross and microscopic features of HCC except for the presence of capsule. These results suggest that DNA ploidy correlates with growth rate of the tumor and it may be a possibly useful prognostic factor in HCCs.

The Epidermal Proliferation and the Number of Langerhans Cells in 7, 12-dimethylbenzanthracene Induced Epidermal Changes.: Chang Soon Han, Young Nyun Park, Kwang Gil Lee, In Joon Choi; Korean J Pathol. 1993;27(6):590-604.

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Abstract PDF: Chemically induced epiderml carcinogenesis is usually divided into two stages, the initiation and promotion. The initiation involves conversion of some epidermal cells into latent neoplastic cells and the promotion is proliferation of the transformed cells. As immunosurveillence is thought to be a host defense against tumors, Langerhans cells, being essential in initiation of local cutaneous immunologic reaction, is suggested to be important in the carcingenesis of the epidermis. This study is attempted to investigate the epidermal proliferative changes in mice induced by application of 12-0-tetradecanoy1-phorbol-13-acetate(TPA) on the skin initiated with 7, 12-dimethylbenzanthracene(DMBA) and its relationship with Langerhans cell. Ninty five male inbred BALB/c mice weighing 20~25 g were divided into five groups; the 33 week-group, the 21 week-group, the 12 week-group and the 4 week-group according to the duration of carcinogen application, and the control group. The carcingen was applied with a brush on the dorsal skin of mice after depilation. Ten days after application of 800 nmole DMBA in 0.4 cc acetone, 20 nmole TPA in 0.4 cc acetone was applied twice per week and the control group was applied with the same amount of acetone for 4 weeks. Animals were sacrificed 3 days after the last application of TPA. One hour before sacrifice, bromodeoxyuridine(BrdU) (1 mg/g) was injected via the tail artert for BrdU stain of S phase cells. A strip of dorsal skin was used for hematoxylineosin stain, immunohistochemical stain for BrdU and la antigen of Langerhans cell, and flow cytometry. The results are as follows: 1. Cellular proliferation, hyperkeratosis and dysplasia of the epidermis were increased in relation to duration of carcingen application. Papillomas were developed 12 weeks after application of the carcingen. 2. BrdU labelling and proliferative indices of the 20 weeks' application group were significantly higher than those of the 12 weeks' application group. The number of Langerhans cell was decreased markedly ater 4 week' application of the carcinogen. 3. All epiedrmal lesions including a case of squamous cell carcinoma were diploidy in flow cytometry. It is thought that disturbance of immunosurveillence, caused by depletion of Langerhans cell, may permit proliferation of epidermal cells. Although abnormal quantitative change of nuclear DNA has not occurred even when the epidermal proliferative activity and dysplastic change were increased markedly, it is thought that the occurrence of structural change of chromosome is remained to be clarified.

Comparative Study

A Comparative Study of PCNA Immunostaining, AgNOR Scores Hormone Receptors and Histology in Human Breast Cancer.: Eun Sook Chang, Kwan Kyu Park; Korean J Pathol. 1993;27(6):605-617.

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Abstract PDF: This study was performed on 50 cases of primary breast cancer removed surgically during the period 1990~1991 and compared the results of four morphologic methods developed for the detection of estrogen receptors, progesterone receptors, monoclonal antibody PCNA immunoreactivity, and the mean number of argyrophilic nucleolar organizer regions(mAgNORs) to ascertain the prognostic significance and also to detect highly malignant heterogenous cancer. To determine the validity of these measurements, a variance analysis was done with Kruskal-Wallis 1-way ANOVA. The results showed that the significant correlation between histologic grade and ER/PR status(P=0.005/P=0.015), the borderline significant correlation between mAgNORs and ER/PR status(P=0.08), and between AgNORs and stage(P=0.07), PCNA has no correlation with ER/PR status(P=0.25), clinical stage and histologic grade. In follow up, four cases of invasive ductal carcinoma with ER/PR, developed early and rapid metastasis within 2 years, three of them were classified as histologic grade 3 and another case was grade 2, whereas two of them were classified ans stage III(+) and the rest were in stage IIB(+) and stage IIA(-). The mAgNOR count of these four cases were ranged from 3.32 to 4.29 which were in the aneuploid category most likely. One of them had rather stormy rapid course with multiple organ metastases resulting death within one year. These results indicated that ductal carcinoma(>2cm size) with ER-/PR- and high mAgNOR level or high PCNA grade, and hihg histologic grade had h highly malignant course, marked by rapidly developing metastases. Thus we concluded that the status of ER/PR alone in tumor tissue is an important information for selecting hormone treatment btu, for the ultimate prognosis, is a weak indicator. Therefore for early detection of such heterogenous tumor, besides ER/PR, AgNORs or PCNA, histologic grade and tumor size are very useful as biological indicators of prognosis. The status ER/PR in combination with these indicators are more accurate and provide better basis on which to base a decision for early implementation of chemotherapy from which to give probably benefit by inhibition of rapid progress.

Original Article

A Morphologic Study on the Bile Duct Changes Induced by Common Bile Duct Ligation in Rats.: Jin Young Jeong, Dae Young Kang, Seung Moo Noh; Korean J Pathol. 1993;27(6):618-629.

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Abstract PDF: In an attempt to elucidate the pathological changes following common bile duct ligation, the present study was undertaken in male Sprague-Dawley rats. Morphologic studies of the livers were performed at 1, 2, 3, 5, 7, 9, 11, 13 and 15 weeks after common bile duct ligation. In an attempt to clarify the relationship between the process of bile duct formation and the nature of primitive cells observable around the primitive biliary structure, light microscopic, immunohistochemical and electron microscopic studies were performed. The results were noted as follows: 1) Light microscopically, proliferation of biliary cells began in the periphery of portal areas and expanded toward hepatic lobules. In severe cases of biliary structure proliferation, hepatocytic cords and classic hepatic lobules were inconspicuous. 2) Immunohistochemically, CK-19 expression was limited to biliary structures in protal areas and proliferated biliary epithelial cells. In the serial sections of paraffin block, proliferated intrahepatic biliary structures were associated with those of portal areas. Some oval cells in the ductular hepatocytes were stained for both CK-19 and MNF 116. 3) Ultrastructurally, the proliferated biliary epithelial cells divided into three patterns: absence of lumen, formation of incomplete lumen, and formation of complete lumen. Furthermore these patterns had spectral continuity of maturation in their structures. 4) In some biliary structures, individual biliary cells pushed the basement membrane toward neighboring tissue with accompanying destruction of basement membranes, patterns of budding. Sometimes these cells and hepatocytes comprised the same lumen. In summary, the results obtained by the present study indicate that proliferated biliary structures may be derived from the preexisting intralobular or portal biliary system.

Case Reports

Peutz-Jeghers Syndrome with Extensive Epithelial Misplacements and Adenomatous and Carcinomatous Transformation: A case report.: Jeong Ja Kwak, So Young Jin, Dong Wha Lee; Korean J Pathol. 1993;27(6):630-637.

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Abstract PDF: Peutz-Jeghers syndrome is an autosomal dominant disease characterized by gastrointestinal ployposis and mucocutaneous melanin pigmentation involving the lip, oral mucosa, digits, palms and soles. The polyps are almost hamartomatous. The relationship of gastrointestinal carcinoma and the Peutz-Jeghers syndrome has been discussed for many years. The question is unsettled whether gastrointestinal carcinoma arise in hamartomatous polyps itself. Recently, there are a few reports that adenomatous and carcinomatous changes were superimposed upon the background of the hamartoma. Occasionally epithelial misplacement of the epithelium is found in the small intestinal polyps. Since the epithelial misplacement may involve submucosa, muscularis propria and serosa, a difficulty of histopathologic differential diagnosis between the epithelial misplacement and invasive adenocarcinoma cause overdiagnosis of cancer in the gastrointestinal polyps of Peutz-Jeghers syndrome. We present a case of Peutz-Jeghers syndrome of 39-year-old woman with multiple gastrointestinal polyps, two of which showed extensive epithelial misplacement even into the pancreas and another one at the colon showed carcinomatous change at the tip portion. Areas of hamartoma, adenoma and in situ carcinoma were noted in this colonic hamartomatous polyp. This case support that adenoma and carcinomatous changes may evolve directly within a hamartomaous polyp itself.

Limb-Body Wall Malformation Complex: Two autopsy cases and its pathogenesis.: Geun Shin Lyu, Nam Hoon Kim, Eun Kyung Hong, Jung Dal Lee; Korean J Pathol. 1993;27(6):638-644.

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Abstract PDF: Limb-body wall malformation complex(LBWC), also know as the amniotic band syndrome, is a poorly defined, sporadic group of congenital anomaly characterized by a collection of protean fetal malformation, deformation and disruption. Accurate diagnosis is often difficult because of its variable presentation pattern and the absence of exactly same case. We report two autopsy cases. One revealed body wall, cardiac, and craniofacial anomalies with anencephaly, and amniotic adhesive band attached to craniofacial defect of the fetal part without evidence of amniotic rupture. The other exibited abdominal wall defect with omphalocele, visceral, postural, and limb anomalies together with neural tube defect in the lumbosacral region. The pathogenesis of this syndrome was discussed in detail.

Duodenal Somatostatinoma: A case report.: Mee Hye Oh, Yi Kyeong Chun, Hye Jae Cho, Jin Ho Lee, Hong Yong Kim, Ill Hyang Ko; Korean J Pathol. 1993;27(6):645-649.

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Abstract PDF: Somatostatinoma is rare endocrine tumor that was first described in 1977 by Ganda et al. and Larsson et al. simultaneously. It seems nonfunctioning at clinical level. But it may present with diabetes, diarrhea, cholelithiasis, steatorrhea, indigestion, hypochlorhydria, and anemia. In contrast with pancreatic somatostatinoma, duodenal somatostatinoma, in general, is clinically silent. Duodenal endocrine tumors show similar histologic pattern. Therefore, the definite diagnosis is performed by immunohistochemistry and electron microscopic examination. We have experienced a case of somatostatinoma of duodenum in a 62-year-old male. He has complained generalized pruritus for one year and jaundice for 2 weeks. Grossly, the mass was a intraluminary protruding, polypoid lesion with focal mucosal erosion at immediately distal to Ampulla of Vater. Histologically, it showed tall, cylindrical cells with distinct cell membranes, having granular cytoplasm and small innocent looking nuclei. No mitosis was seen. The tumor cells were arraged in small solid groups and trabeculae, separated by fibrovascular stroma. Immunohistochemically, the tumor cells were strongly positive with somatostatin and negative with several other hormonal and neuroendocrine markers. Ultrastructurally, the cytoplasm contains numerous, homogeneous low electron dense secretory granules, which are essentially similar to those seen in normal delta cells.

Malignant Melanoma Arising in Giant Congenital Melanocytic Nevus: A case report.: Jung Sun Kim, Sang Yong Song, Kye Yong Song, Je G Chi; Korean J Pathol. 1993;27(6):650-655.

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Abstract PDF: Giant congenital melanocytic nevus is found in 0.1% of live born infants. If present, this lesion has a 6.3% chance to develop malignant melanoma. We report such a case in a 22-year-old woman who had multiple pigmented skin lesions since birth. Rapidly growing masses were recently detected in the 19 cm-sized occipital pigmented lesion. Removed scalp lesion revealed yellowish white lobulated soft nodules in the background of pigmented nevus. Microscopically, the nodules consisted of epithelioid cells with prominent nucleoli, and pleomorphic cells including signetring cells. These cells seldom contained melanin pigment. There were metastatic aggregates of tumor cells in the cervical lymph node, which were reminiscent of germinal centers of lymph nodes. S-100 protein immunostaining was helpful to distinguish them. Incidentally, focally scattered pigmented spindle cells were seen in the capsule of a lymph node

Breast Mass as a Manifestation of Ectopic Paragonimiasis: A case report.: Yung Suk Lee, Seung Yeon Ha, Hyun I Cho, Han Kyeom Kim, Jung Won Bae, In Sun Kim; Korean J Pathol. 1993;27(6):656-658.

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Abstract PDF: Paragonimus westermani can cause extrapulmonary parasitism in various sites such as abdominal organ, brain, eye, periorbital tissue, heart and pericardium, mediastinum, and subcutaneous tissue. We experienced a case of subcutaneous paragonimiasis involving the breast. The lesion exhibited chronic granuloma with scattered eggs of paragonimus westermani. The adult worm was not found within the lesion which should be disintegrated a year ago by praziquantel treatment. This relatively rare involvement of ectopic paragonimiasis should be differentiated from breast malignancy.

Bednar Tumor: A case report with discusion of histogenesis.: Mee Soo Chang, Kye Yong Song; Korean J Pathol. 1993;27(6):659-662.

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Abstract PDF: The authors examined a case of Bednar tumor(pigmented dermatofibrosarcoma protuberans). A 35-year-old woman presented with protruding black back mass 3x2.5 cm in surface dimension and 1 cm in heigh, which has grown slowly for 30 years. Microscopically the lesion is characterized by spindle cells arranged in storiform pattern and admixed with melanin containing dendritic cells. Ultrastructural study revealed three cell populations: (1) cells resembling fibroblasts, (2) cells resembling perineural cells, and (3) melanocytes. The immunohistochemical study using S-100 protein alpha-1 antitrypsin, antichymotrypsin and EMA revealed negative reaction in spindle fibroblastic tumor cells. The histogenesis of this neoplasm remains controversial; neuroectoderm, perineural cell, fibroblast and histiocyte. Through the ultrastructural study and immunohistochemical study, the authors suggested that Bedner tumor arises form the neuroectoderm with bidirectional to perineural and fibroblastic differentiation.

Squamous Metaplasia in Tubular Adenoma of Sigmoid Colon: A case report.: Soo Min Kang, Weon Seo Park, Woo Ho Kim, Yong Il Kim; Korean J Pathol. 1993;27(6):663-665.

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Abstract PDF: The occurrence of squamous metaplasia(morule) in colorectal mucosa and adenocarcinoma, althrough rare, has been well documented. In contrast, very little mention has been given to mature squamous cells seen in colorectal polyps or adenomas. A 42-year-old woman presented with a 2-month history of diarrhea and melena. Proctosigmoidoscopy revealed a 4 cm-sized polypoid tumor 20 cm above the anal verge. Colonoscopic biopsy showed tubular adenoma, and a segmental resection of sigmoid colon was done. Microscopically, the tumor was c classical tubular adenoma containing multiple solid nests of squamous cells scattered only in the neoplasm; the squamous nests were generally small, and some showed direct continuity with adenomatous glands. The squamous cells were keratinizing and had regular nuclei with no mitotic activity. The importance of this phenomenon lies in its pathologic recognition, and the findings suggest that awareness of this rare occurrence in colorectal polyps should avert such overdiagnosis, and consequently prevents unnecessary radical surgery.

Pleomorphic Xanthoastrocytoma: A Case Report.: Seong Ki Min, Dong Wook Kang, Kyu Sang Song, Dae Young Kang, Seong Ho Kim; Korean J Pathol. 1993;27(6):666-669.

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Abstract PDF: Pleomorphic xanthoastrocytoma is histologically characterized by marked cellular pleomorphism of lipid-laden neoplastic astrocytes and bizarre giant cells showing mitotic figures and high cellularity. Inspite of its ominous-looking microscopic features, howerver, the prognosis is usually favorable. This tumor develops mainly in the supratentorial area of young people and frequently involves the leptomeninges. We experienced a case of pleomorphic xanthoastrocytoma in 18 year-old-male. In addition to the cellular pleomophism, the prominent reticulin fibers surround the individual tumor cells or the tumor cells nests. Immunohistochemical staining and electron microscopy revealed glial fibrillary acidic protein(GFAP) expression and pericytoplasmic basal lamina in the tumor cells.

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