Journal of Pathology and Translational Medicine

Volume 34(3); March 2000

Original Articles

Pulsating Magnetic Field Effects on in vitro Culture of Human Osteogenic Sarcoma Cell Lines.: Hyo Sook Shin, Jin Young Lee, Suk Keun Lee, Sang Chul Park, Je G Chi; Korean J Pathol. 2000;34(3):169-180.

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Abstract PDF: In order to elucidate the biological effects of pulsating magnetic field in in vitro culture system we designed a pulsating magnetic apparatus using 120 Hertz, 24 Volt direct current. It can generate 63~225 Gauss in the experimental area of 90 mm petri dish, and has little thermal effect on the culture media in 37.5oC, 5% CO2. Human osteogenic sarcoma (HOS) cells were cultured in the pulsating magnetic field and the nuclear changes of cultured cells were observed routinely by hematoxylin staining, and apoptotic change was detected by ApopTag staining using both peroxidase and fluorescein labelings. Compared to the control group which formed well organized whorling pattern of HOS cell line in 3 days culture, the HOS cells cultured in the pulsating magnetic field for 12 hours or 24 hours grew irregularly and showed increased number of apoptotic cells. When the flow of pulsating magnetic field was interrupted by insertion of strong permanent magnetic bar (1000 Gauss, 5530 mm) beneath the petri dish during in vitro culture, the area of sparse pulsating magnetic field showed active proliferation and aggregation of HOS cells even in 24 hour exposure group. These data suggest that the pulsating magnetic field may play a role in inducing growth retardation and apoptosis of HOS cells. Furthermore, the hazardous effects of pulsating magnetic field can be lessened or nullified by the interruption of pulsating magnetic field with a strong permanent magnetic bar.

Three Dimensional Reconstitution of Oral Mucosal Keratinocytes and Its Biologic Characteristics.: In Ho Cha, Jong In Yook, Young Sook Son, Eun Ha Lee, So Young Jeong, Kyung Joo Kim, Jin Kim; Korean J Pathol. 2000;34(3):181-189.

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Abstract PDF: The purposes of this study were to develop an in vitro co-culture model of epithelial tissue with dermal equivalent, cultured at an air-liquid interface, and to evaluate the effects of extracellular matrix and concentration of calcium and fetal bovine serum in medium to find optimized culture condition. Oral keratinizing epithelial cells in monolayer culture were grown in Mitomycin-treated 3T3 feeder. Primary cultured oral epithelial cells were reconstituted onto the dermal equivalents consisting of 3T3 fibroblast and type I collagen, and co-culture was grown at the air-liquid interface. The histomorphological development of reconstituted oral epithelium in vitro for 21 days revealed 10~12 layered statified epithelium, closely similar to the parakeratinized gingival epithelium. Neither laminin nor type IV collagen was able to induce keratinocyte differentiation. But a mixture of laminin and type IV collagen induced well-polarized keratinizing tissue with anchoring structure of basal cells. When the reconstituted oral epithelium was incubated in 1.0% and 0.5% serum-containing medium, the granular cell layers with orthokeratinization developed. The reconstituted epidermis generated in serum-free keratinocyte growth medium (KGM)-containing pituitary extract showed features of incomplete differentiation. The present study shows that the dermal equivalents containing fibroblasts will support epidermal morphogenesis and differentiation. And these results suggest that extracellular matrix and calcium concentration are important factors during the reconstitution of keratinizing epithelium in vitro.

VEGF Expression and Microvessel Density in Oral Squamous Cell Carcinomas.: Ji Jun Lim, Sam Pyo Hong, Jae Il Lee, Seong Doo Hong, Chang Yun Lim; Korean J Pathol. 2000;34(3):190-198.

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Abstract PDF: Angiogenesis is an essential process in tumor growth and metastasis. VEGF has been considered a leading candidate inducing tumor angiogenesis. VEGF expression was significantly correlated with clinical stage, lymph node matastasis, and prognosis of cancers of various parts of body. However, little has been known about the correlation between VEGF expression and clinicopathologic parameters in oral squamous cell carcinoma. The aim of this study was to correlate VEGF expression with the clinicopathological parameters and microvessel density. Forty six oral squamous cell carcinomas were analyzed using immunohistochemical method with primary antibodies to VEGF and CD31. VEGF expression was detected in 33 (71.7%) of the 46 cases. The microvessel density was significantly correlated with VEGF expression (P=0.002). There was no correlation between microvessel density and tumour size, clinical stage, and lymph node metastasis, respectively. VEGF expression did not correlate with the histological grade of tumour differentiation, tumour size, and clinical stages. The VEGF-positive rate seemed to be higher in patients with cervical lymph nodal metastasis than in those without it, but it was not statistically significant. In conclusion, the overexpression of VEGF in the oral squamous cell carcinoma seemed to be associated with a more aggressive course of the disease. Further study is necessary to define the role of VEGF in oral squamous cell carcinoma.

Expression of Insulin-like Growth Factor-I-Receptor in Colorectal Adenomas and Carcinomas.: Young Chae Chu, Hye Seung Han, Jee Young Han, Joon Mee Kim, Young Bae Kim, Tae Sook Hwang; Korean J Pathol. 2000;34(3):199-207.

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Abstract PDF: The activation of the insulin-like growth factor-I-receptor system (IGF-IR) has recently emerged as critical events in transformation and tumorigenicity of several human tumors. In this study we investigated the expression of IGF-IR in 33 colorectal adenomas, 88 primary colorectal carcinomas, and 30 normal colonic mucosa adjacent to the carcinoma. Immunohistochemical staining (IHC) for IGF-IR was performed on paraffin embedded sections using an anti-IGF-IR rabbit polyclonal antibody. IHC stains for IGF-IR were scored using a semiquantitative scoring system. The relationship of IGF-IR staining to clinicopathologic variables and proliferating cell nuclear antigen (PCNA) staining was also analysed. The mean IHC scores for IGF-IR of normal glands, adenoma, intramucosal carcinoma, node-negative carcinoma, and node-positive carcinoma were 0.41 0.96, 0.76 1.23, 2.0 1.48, 2.83 2.0 and 5.93 1.58, respectively. These scores for each category were statistically significant except between normal glands and adenoma and between intramucosal carcinoma and node-negative carcinomas. The mean PCNA indexes of normal glands, adenoma, intramucosal carcinoma, node-negative carcinoma, and node-positive carcinoma were 2.48 2.60, 6.94 11.03, 27.21 11.42, 43.36 9.9 and 57.60 10.01, respectively. The PCNA index for each category was statistically significant except between normal and adenoma. IGF-IR scores and PCNA indexes were higher with tumor progression and also correlated each other (sr=0.65, p=0.0001). Higher IGF-IR scores and PCNA indexes were seen in tumors with advanced stage, infiltrative growth pattern, poor differentiation, nerve invasion, lymphovascular invasion, and moderate fibrosis. Our results suggest that IGF-IR plays an important role in tumorigenicity and tumor progression.

Microvillous Inclusion Disease: An analysis of 4 cases.: Daesu Kim, Nam Seon Beck, Yeon Lim Suh; Korean J Pathol. 2000;34(3):208-213.

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Abstract PDF: Microvillous inclusion disease (MID) or congenital microvillous atrophy is a rare cause of intractable, persistent secretory diarrhea shortly after birth or at birth. The prognosis of MID is very poor and most of the reported patients have died within 6.5 years. Diagnosis is based on the ultrastructural demonstration of intracytoplasmic inclusion of microvilli. There have been several cases reported in the literature. To evaluate the clinicopathologic features of MID, we have reviewed 4 cases of MID which was confirmed by the ultrastructural study of the duodenal biopsy. All patients suffered from life-threatening diarrhea since birth. Many other radiologic or laboratory findings were of no value in the evaluation of causative agents. One of the patients died of the disease and remaining patients have been alive on intravenous fluids or total parenteral nutrition. Histological findings of all cases were similar and characterized by varying degrees of mucosal atrophy and chronic inflammatory cell infiltration in the duodenal mucosa. PAS stain revealed a discontinuous brush border over the atrophic villous surface with or without small vacuoles in the cytoplasm of the surface epithelium. Ultrastructural changes were found mainly in the surface epithelium of the duodenal mucosa and characterized by the presence of membrane bound inclusions lined by intact or degenerating microvilli, as well as degeneration of surface epithelial cells with loss of microvilli, or with sparse, short microvilli.

Protective Effects of Captopril in Radiation-Induced Renal Injury in Rats.: Ji Yeon Bae, Eun Sook Chang, Ok Bae Kim; Korean J Pathol. 2000;34(3):214-224.

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Abstract PDF: The angiotensin I converting enzyme inhibitor (Captopril) has recently been studied extensively in various experimental models of radiation injury and has proven its protective effects in various organs, such as the lungs, gastrointestinal tract, and kidneys. Twenty-three Sprague-Dawley rats were divided into experimental and control group. The experimental group was divided into two large groups: the first one received a single dose of 18 Gy irradiation from an electron beam on the local field of the kidney region only, and the second group received captopril per oral continuously after the same doses of irradiation. The second experimental group was divided into four subgroups by captopril doses: 62.5 mg/l, 125 mg/l, 250 mg/l, and 500 mg/l, respectively. On light and electron microscopy, the kidneys of the irradiated rats with no captopril treatment showed diffuse glomerular contraction, congestion with occlusion and focal necrosis of the endothelial, and mesangial cells. The tubules showed ballooning degeneration, desquamation, and diffuse coagulation necrosis. Captopril treated rats, especially those given a high dose (more than 250 gm/l), revealed a marked reduction of the tubular and glomerular injuries. There was a statistically significant difference in the degree of renal injury among the experimental groups (p<0.05). The result of this study suggests that an administration of high dose captopril might prevent radiation-induced renal injury, especially in the early post-irradiation period.

Anticancer Effect and Apoptosis of All-trans-retinoic Acid on the Human Ovarian Epithelial Carcinoma Cell Lines.: Jee Young Han, Woo Hee Jung, Tai Seung Kim; Korean J Pathol. 2000;34(3):225-234.

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Abstract PDF: Ovarian carcinoma is a serious disease in women. Some reports revealed all-trans-retinoic acid (tRA) inhibited the proliferation of ovarian carcinoma cell lines and induced apoptosis. The aim of this study was to evaluate the anticancer and apoptotic effects of tRA and the expression of the retinoic acid receptor (RAR) alpha, beta, gamma, p53, bcl-2, and c-myc genes on the ovarian carcinoma cell lines, NIH OVCAR3 and SKOV3. In both cell lines, the proliferation of tumor cells was inhibited and characteristic morphologic patterns of apoptosis were shown after treatment of tRA. The number of apoptotic cells and the percentage of apoptosis were increased after treatment of tRA. The gel electrophoresis revealed the DNA ladder pattern in the NIH OVCAR3. Gene expressions were observed using northern blotting. There was no RARalpha expression in both cell lines. In NIH OVCAR3, there was no changes in the expression of RARbeta and bcl-2 gene. The RARgamma gene expression of tRA treated group was slightly increased, but p53 gene expression was decreased, and c-myc gene was not expressed. In SKOV3, the expressions of RARbeta, gamma, and p53 genes were increased and that of bcl-2 was decreased in the tRA treated group. There was no change in c-myc gene expression. These results suggest tRA has anticancer and apoptotic effect on both ovarian carcinoma cell lines. RARbeta, RARgamma, bcl-2, and p53 gene expressions were correlated with these effects of tRA on SKOV3 but not on NIH OVCAR3.

An Imported Case of Intestinal Capillariasis Presenting as Protein-Losing Enteropathy.: Youngmee Kwon, Hwoon Yong Jung, Hyun Kwon Ha, Inchul Lee; Korean J Pathol. 2000;34(3):235-238.

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Abstract PDF: Intestinal capillariasis is caused by a small trichurid nematode, Capillaria philippinensis, which infects freshwater fish as intermediate hosts and subsequently fish eating birds as definitive hosts. It has occurred in areas such as the Philippines and Thailand where people eat raw fish. We report an imported case of intestinal capillariasis in a 31-year-old Korean man who lived in Saipan for ten years. He suffered from diarrhea with weight loss, abdominal distension, and pitting edema in the lower extremities for two years. He ate raw freshwater fish in the Saipan 2 years ago. The diagnosis was confirmed by jejunal biopsy. Flat mucosal surface without villi contained sections of numerous round worms with stichosome, larvae, and eggs which were identified as Capillaria philippinensis. The patient was successfully treated with albendazole.

Juvenile Hyaline Fibromatosis in an Adult.: Young A Kim, Seoung Wan Chae, Chong Jai Kim, Je G Chi; Korean J Pathol. 2000;34(3):239-242.

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Abstract PDF: Juvenile hyaline fibromatosis is a rare disorder probably inherited as an autosomal recessive trait. It is characterized by multiple slowly growing subcutaneous nodules, hypertrophy of gingiva, flexion contracture, and radiolucent bone destruction. The histological features of the tumor-like lesions are characterized by the deposition of amorphous hyaline material in which spindle shaped cells are embedded. We report a case of juvenile hyaline fibromatosis in a 26 year-old-woman. She had multiple subcutaneous nodules in scalp, ear, forearms, right knee, and back. Surgical excision of the tumors in the scalp and ear was done. The largest one measured 13 9 6 cm, and had homogeneous, grayish yellow cut surface with calcification. Light microscopic examination showed abundant eosinophilic hyaline material with extensive calcification and metaplastic bone formation. Spindle cells were rarely observed at the periphery of the tumor. Hyaline matrix was PAS positive, diastase resistant, and alcian blue negative. Scattered spindle cells were positive for vimentin but negative for S-100 protein and smooth muscle actin. There were many reports regarding early lesions of juvenile hyaline fibromatosis; however in this patient, tumor existed for more than 20 years and the histology was somewhat different from the early lesions reported in the literature.

Case Report

Hemimegalencephaly: A case report.: Youn Soo Lee, Eun Sun Jung, Sun Moo Kim, Byung Kee Kim; Korean J Pathol. 2000;34(3):243-247.

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Abstract PDF: Hemimegalencephaly is an uncommon sporadic congenital malformation of the central nervous system characterized by enlargement of one cerebral hemisphere with cranial asymmetry, hemiparesis, epilepsy, and mental retardation. This report deals with a 12-month-old girl with intractable seizure and developmental delay, who revealed hemispheric asymmetry and enlarged right cerebral hemisphere. MRI scan revealed asymmetric enlargement of right cerebral hemisphere with lissencephaly and multifocal calcification. She underwent right frontal lobectomy and parietal cortical resection, but died of complication of surgery. Histological features included loss of cortical lamination, large atypical neurons, neuronal heterotopia, and astrocytosis with dystrophic calcification. The heterotopic neurons in the white matter were suggestive of aberrant neuronal migration.

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