Journal of Pathology and Translational Medicine

Volume 33(11); November 1999

Original Articles

Factors Influencing Regeneration of Calvarial Defects in Rats.: Sung Chul Lim, Young Sook Kim; Korean J Pathol. 1999;33(11):999-1008.

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Abstract PDF: An experimental study was done to evaluate factors influencing guided regeneration of bone in standardized calvarial bony defect. An 8 mm circular transosseous calvarial bony defect was made. Various material such as demineralized freeze-dried bone (DFDB), BioMesh , Millipore filter and its combination was placed in the bony defect. A sequential histopathologic, histochemical, immunohistochemical, and histomorphometric studies were done on the guided bone regeneration in the calvarial bony defect. Bone formation was sigificantly enhanced when the DFDB was retained within the bony defect with a protective bioabsorbable membrane. Inframembranous DFDB-filling was required to prevent collapse of the membrane and preserve spaces for bone regeneration. The bioabsorbable membrane should presumably remain intact for longer than at least 5 weeks to facilitate bone regeneration. The new bone formation was dependent on the barrier-effect (preserving secluded spaces) and inflammation-inducing property of membrane, and guiding bone regeneration of the grafts. Macrophages recruited by grafts were partly involved in decrease of bone regeneration via the sequential events of release of fibronectin, chemotactic effect of the fibronectin to fibroblasts, and collagen lay-down.

Immunohistochemical Analysis of Transforming Growth Factor (TGF)-beta1 and TGF-beta Receptor II and Quantitative Analysis of TGF-beta1 mRNA during Multistep Hepatocarcinogenesis Induced by Diethylnitrosamine in Sprague-Dawley Rats.: Mee Yon Cho, Ju Han Lee, Yong Koo Kang, Nam Hee Won; Korean J Pathol. 1999;33(11):1009-1023.

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Abstract PDF: Transforming growth factor (TGF)-beta1 plays an important role in hepatocarcinogenesis and has been described as a useful tumor marker and one of the poor prognostic indicators in patients with hepatocellular carcinoma (HCC). To investigate the role and cellular localization of TGF-beta1 during multistep hepatocarcinogenesis we performed a quantitative analysis of TGF-beta1 mRNA and immunohistochemical expression of TGF-beta1 and TGF-beta receptor II (TGF-betarII) in female Sprague-Dawley rats. The experimental groups included neoplastic lesions produced by Solt-Farber's protocol, regenerating liver after partial hepatectomy, and normal control. Quantitative change of TGF-beta1 mRNA was analysed by competitive reverse-transcription polymerase chain reaction (RT-PCR). TGF-beta1 protein and TGF-betarII expression were evaluated by immunohistochemical stain. The discrete tumor nodules were detected on 14th day and then increased in number and size. Three HCCs were induced on 8th or 9th month. RT-PCR demonstrated TGF-beta1 mRNA band in all examples of the normal and regenerating liver, nodules and HCCs. Competitive RT-PCR displayed higher TGF-beta1 mRNA in nodules, HCCs and regenerating liver than in normal controls. Hepatocytes from control and regenerating livers showed weak immunoreactivity for TGF-beta1. In contrast, the cytoplasm of hepatocytes of nodules in 7th, 8th and 9th month and HCCs were intensely stained for TGF-beta1. Some sinusoidal cells showed immunoreactivity for TGF-beta1 in all experimental groups. In early phase of carcinogenesis, the cytoplasm of hepatocytes in liver of 12h, 1d and 3d showed transiently increased immunoreactivity for TGF-beta1 and The immunoreactivity decreased thereafter. TGF-beta1 mRNA was also detected in the neoplastic hepatocytes by in-situ hybridization. Although TGF-betarII expression was correlated with TGF-beta1 immunoreactivity during early phase of carcinogenesis, hepatocytes in most nodules in 7th, 8th, 9th month and carcinomas showed decreased or little immunoreactivity for TGF-betarII. Based on the above results, it is concluded that TGF-beta1 expression increases not only in precancerous nodules but also in HCCs and its increase seems to be correlated with decrease or loss of TGF-betarII expression although its mechanism remains unclear. Hepatocytes may be a major cellular source of TGF-beta1 during hepatocarcinogenesis.

Talc Deposition in Lipoma: A Pathologic and Mineralogic Study.: Jung Ran Kim, Hyun Sul Lim, Hae Kwan Cheong, Ji Yong Kim, Kiyoshi Sakai, Naomi Hisanaga; Korean J Pathol. 1999;33(11):1024-1032.

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Abstract PDF: Three cases of lipomas associated with heavy talc deposits are reported in local inhabitants near a fiber glass factory. Pathologic and mineralogical analysis by polarizing microscopy, scanning and transmission electron microscopy with energy dispersive X-ray analysis and X-ray diffraction of mass were done. Simultaneously, we performed an epidemiological survey and a mineralogical study of disposed fiber glass from waste site and ground water. All tumors consisted of mature fat tissue containing an abundant birefringent talc fibers and particles under polarized light. The concentrations of the fibers were 494.7, 6.7 and 50.7 million fibers per gram of dry tissue. The fibers were needlelike with rectangular fractured ends, up to 17 micrometer in length and 0.5 micrometer in diameter. EDX and X-ray diffraction analysis of the fibers showed that 71 to 100% of the fibers were magnesium silicate, talc. We also identified magnesium silicate fibers in the fiber glass from the waste site and in the ground water which were similar to talc fibers in lipomas,. We concluded that fibrous talc, a component of fiberglass waste, might be associated with the development of lipomas via unknown exposure route.

Diagnostic Usefulness of Monoclonal Antibody for T Lymphoblastic Lymphoma/Acute Lymphoblastic Leukemia-Specific JL1 Antigen in Paraffin Embedded Tissue.: Chan Sik Park, Seong Hoe Park; Korean J Pathol. 1999;33(11):1033-1038.

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Abstract PDF: JL1 is a novel human thymocyte differentiation antigen, which is exclusively expressed by double positive (CD4+ CD8+) cortical thymocytes. We previously reported that the JL1 antigen was selectively expressed on the surface of acute lymphoblastic leukemia cells. T-Lymphoblastic lymphoma/acute lymphoblastic leukemia (T-LBL/ALL), the 6th prevalent lymphoma in Korea, is composed of immature neoplastic lymphoid cells and shows a rapid response to appropriate treatment. Early and precise diagnosis of LBL/ALL is crucial. Light microscopic distinction of LBL/ALL from other non-Hodgkin's lymphomas can sometimes be difficult and is aided by immunophenotypic studies. This study is designed to investigate the diagnostic utility of anti-JL1 monoclonal antibody (YG5) for LBL/ALL in formalin fixed, paraffin embedded tissue. We collected 25 cases of LBL/ALL (18 T-cell, 5 B-cell and 2 undetermined lineage) from 1993 through 1998. We confirmed the diagnosis using morphologic and immunophenotypic data. Strong JL1 expression along cell membrane was observed in 16 out of 18 T-LBL/ALL cases (89%). In 28 cases of other types of lymphomas of including 7 cases of non-T LBL/ALL and 14 cases of small round cell tumors, no JL1 expression was identified. These results show that the immunostaining for JL1 using YG5 on paraffin embedded sections can be useful for the specific diagnosis of T-LBL/ALL in routine diagnostic service.

Expression of Epidermal Growth Factor Related Peptides, EGF-R, and c-erbB-2 and Their Relationship with the Prognostic Factors in Gastric Carcinoma.: Joo Heon Kim, Jin Wook Lee, Woo Sung Moon, Myoung Jae Kang, Dong Geun Lee; Korean J Pathol. 1999;33(11):1039-1046.

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Abstract PDF: Recent investigations have revealed that autocrine growth factors and their receptors are closely related and play an important role in controlling cancer cell growth. We performed an immunohistochemical study on the expression of epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), epidermal growth factor receptor (EGF-R), c-erbB-2, and PCNA labelling index in 60 cases of human gastric carcinomas. TGF-alpha was detected in 38 cases (63.3%), EGF in 26 cases (43.3%), EGF-R in 44 cases (73.3%), and c-erbB-2 in 18 cases (30%). These growth factors, EGF-R and c-erbB-2, were found more often in advanced gastric cancers. The PCNA labeling index was significantly higher in tumors with the expression of EGF-R or c-erbB-2. Tumors with simultaneous expression of EGF, TGF-alpha, EGF-R and c-erbB-2 was associated with a high PCNA labeling index. A correlation was observed between the synchronous expression of growth factors and its receptors and histological differentiation. The results suggest that the expression of EGF, TGF-alpha, EGF-R and c-erbB-2 are closely related and plays an important role in the growth and progression of human gastric carcinoma.

Type IV Collagen mRNA Expression in Human Membranous Nephropathy.: Tae Sook Kim, Jung Yeon Kim, Hye Kyoung Hong, Hyun Soon Lee; Korean J Pathol. 1999;33(11):1047-1054.

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Abstract PDF: Human membranous nephropathy (MN) is morphologically characterized by subepithelial immune complex deposits and progressive thickening of glomerular basement membranes (GBM). Studies have suggested that the enhanced secretion of classical and novel type IV collagen chains in MN contributes to spike formation and the novel type IV collagen chain is particularly related to thickening of GBM. It is unclear whether the increased accumulation of extracellular matrix (ECM) proteins in GBM is due to the increased mRNA expression for type IV collagen in glomerular visceral epithelial cells (GECs). To answer this question, we analyzed seven renal biopsies of patients with idiopathic MN using in situ hybridization. In MN, the number of GECs expressing mRNA for alpha1(IV) collagen was 2.82+/-1.80/glomerular cross section (gcs), and the number expressing mRNA for alpha4(IV) collagen was 8.42+/-2.85/gcs. The number of GECs expressing mRNA for alpha4(IV) collagen was significantly larger than that of alpha1(IV) collagen mRNA. The expression of mRNA for these ECM proteins in normal controls was negligible. These results suggest that subepithelial immune complexes stimulate the gene expression of alpha1(IV) collagen and alpha4(IV) collagen in glomerular GECs which, in turn, increase the secretion of ECM proteins and contribute to the thickening of GBM in MN.

Tumor Angiogenesis in Renal Cell Carcinoma.: Ji Shin Lee, Jong Jae Jung, Chang Soo Park; Korean J Pathol. 1999;33(11):1055-1060.

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Abstract PDF: Angiogenesis is essential for the growth of solid tumors. Microvessel counts, which represent a measure of tumor angiogenesis, have been correlated with the overall survival of patients with a variety of malignancies. However, the significance of angiogenesis in renal cell carcinoma remains controversial. To determine whether angiogenesis correlates with prognosis of patients with renal cell carcinoma, we counted the microvessels within the primary tumors and compared their numbers with patients' prognosis. Tumor specimens from 42 patients were investigated. Microvessels were stained with anti-CD34 and anti-factor VIII-related antigen monoclonal antibodies. Significant correlation between microvessel counts for two antibodies was observed (r=0.875, p<0.01), although microvessel counts for CD34 were approximately two times higher. Microvessel counts were higher in clear cell than in non-clear cell carcinoma (p<0.05). These results suggest that immunostaining with anti-CD34 antibody may provide a more sensitive and accurate measure of tumor angiogenesis. There was no correlation between microvessel counts and nuclear grade, or TNM stage. In univariate analyses, nuclear grade and TNM stage were significantly associated with patient survival (p<0.01). But further studies on tumor angiogenesis of renal cell carcinoma are needed before it can be adopted as a prognostic marker.

Correlation of Clinical Stage and Presumptive Prognostic Factors in Renal Cell Carcinoma.: Jin Ye Yoo, Hye Jae Cho; Korean J Pathol. 1999;33(11):1061-1066.

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Abstract PDF: Renal cell carcinoma is the most common primary cancer of the kidney. The tumor stage is a reliable prognostic marker in renal cell carcinoma which is significantly associated with patient survival. But assessment of other prognostic factors has produced varying and often conflicting results. We reevaluated the significance of varied prognostic parameters in 33 cases of renal cell carcinoma; clinical stage, cell type, histologic pattern, DNA ploidy, Ki-67 labeling index, and bcl-2 oncoprotein expression. We could not statistically prove that DNA ploidy and bcl-2 expression were related to any examined parameters. Cell type was not related to clinical stage nor nuclear grade but there was a significant correlation (p=0.002) between cell type and histologic pattern. Nuclear grade (p=0.007) and Ki-67 labeling index (p=0.036) were significantly related to clinical stage, suggesting their value as complementary prognostic markers for renal cell carcinoma.

Clinicopathologic Features of Prostatic Adenocarcinoma: A Study of 58 Radical Prostatectomy Specimens.: Tae Jin Lee, Mi Sun Choe, Choung Soo Kim, Jae Y Ro; Korean J Pathol. 1999;33(11):1067-1075.

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Abstract PDF: Prostatic carcinoma is the most commonly diagnosed cancer in the United States in men. Prostatic carcinoma in Korean men is uncommon and accounts for less than 1% of total cancer: however, the incidence of prostatic carcinoma is continuously increasing. Several clinicopathologic parameters including stage, Gleason score, and serum PSA level have been widely accepted as well established prognostic factors. To study the clinicopathologic features of prostatic carcinoma in Korean men, 58 cases of prostatic carcinoma, which were diagnosed on radical prostatectony specimens at Asan Medical Center from Jan. 1993 to June 1998 (1993; 3 cases, 1994; 3, 1995; 6, 1996; 12, 1997; 24, 1998; 10), were evaluated. The prostatic carcinomas were divided into three groups according to Gleason score. Tumors with Gleason score 6 or lower, 7, and 8 to 10 were categorized as low-grade, intermediate-grade and high-grade tumor, respectively. The overall mean age of the patients was 62.6 years (range, 4 6~76 years); mean age was 65.4, 62.8 and 61.1 in low, intermediate and high-grade tumor, respectively (p>0.05). The overall mean serum PSA level was 38.6 ng/ml (range, 0.3~276.0 ng/ml); mean serum PSA level was 17.0, 29.0 and 60.9 ng/ml in low, intermediate and high-grade tumor, respectively (p=0.002). The mean T stage was 2.3, 2.4 and 3.1 in low, intermediate and high-grade tumor (p=0.001). The percentage of positive resection margin was 33.3, 50.0 and 91.0 % in low, intermediate and high-grade tumor (p=0.001). The overall presence of prostatic intraepithelial neoplasia (PIN) was 79.3 %; percentage of the presence of PIN was 100, 79.2 and 68.2 % in low, intermediate and high-grade tumor (p>0.05). As reported in the literature, our results indicated that Gleason score was a good predictor of stage and prognosis. The higher Gleason score, the more cases were with positive surgical margins, advanced pathologic stage, and high serum PSA level (p<0.05).

Expression of Biologic Markers and DNA Ploidy Analysis in Atypical Ductal Hyperplasia and Ductal Carcinoma in Situ of the Breast.: Hee Jung Kim, Woo Hee Jung, Hyeon Joo Jeong, Hy De Lee; Korean J Pathol. 1999;33(11):1076-1089.

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Abstract PDF: Status of margins and the size of the lesion are independent prognostic factors of ductal carcinoma in situ (DCIS). Histologic grading of DCIS and expression of biologic marker also appear to act as prognostic factors. However, DNA ploidy analysis using flow cytometry in the DCIS and atypical ductal hyperplasia (ADH) has been rarely reported, and the biologic behavior of ADH is unknown. We performed immunohistochemical staining and DNA ploidy analysis using flow cytometry on 45 cases of pure DCIS without microinvasion and 34 cases of ADH to compare the expression of biologic markers and DNA ploidy patterns according to the histologic grade of DCIS, to evaluate the usefulness of the Van Nuys classification, and to investigate the biologic behavior of ADH and low grade DCIS. A total of 41.9% of DCIS and 32.1% of ADH were detected mammographically in asymptomatic patients. The most common subtype of the high grade DCIS was comedo type (56.3%), while the low and intermediate grade DCIS were cribriform type. Expression of ER, c-erbB-2 and Ki-67 proliferative index (PI) was significantly associated with nuclear grade and histologic grade of DCIS. Expression of c-erbB-2 was also significantly correlated with presence of necrosis. In low grade DCIS, Ki-67 PI was significantly higher than ADH. A total of 63.6% of DCIS and 70% of ADH were diploidy and 15.9% of DCIS was aneuploidy. There was no aneuploidy in ADH. No significant association was noted between DNA ploidy and histologic grade or nuclear grade. However, in high grade DCIS, the frequency of aneuploidy was high. In conclusion, histologic grading of DCIS employing nuclear grade and necrosis is a useful tool accounting for biologic behavior. High grade DCIS and comedo DCIS impart aggressive biologic behavior and suggest a higher possibility of local recurrence or progression to invasive carcinoma. In the differential diagnosis of ADH and low grade DCIS, the use of Ki-67 PI and DNA ploidy analysis by flow cytometry will be helpful for accurate diagnosis and prediction of biologic behavior.

Case Reports

Williams Syndrome in an Infant An autopsy case report .: Jeong Hae Kie, Se Hoon Kim, Jae Young Choi, Sang Ho Cho; Korean J Pathol. 1999;33(11):1090-1093.

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Abstract PDF: Williams syndrome is a congenital disorder characterized by mental retardation, loquacious personalities, dysmorphic face, and vascular and valvular abnormalities. The etiology of this syndrome was one allelic loss of elastin gene, exhibiting a submicroscopic deletion, at 7q11.23. Sudden death is an infrequently recognized complication. The mechanism of sudden death is explained by myocardial ischemia, decreased cardiac output, and arrhythmia by anatomical abnormality of coronary artery stenosis and severe biventricular outflow tract obstruction. We report an autopsy case of a 80 day-old male with Williams syndrome. Five days before admission, cardiac murmur was detected incidentally on ascultation at a local clinic during a visit for vaccination. He was transferred to our hospital and cardiac catheterization was done. He died suddenly next day. Postmortem examination revealed a dysmorphic face and multiple cardiovascular abnormalities including supravalvular aortic stenosis with narrowed coronary artery ostia, supravalvular pulmonic stenosis, secundum type of atrial septal defect, right ventricular hypertrophy, and renal artery stenosis. Histologically, aorta and pulmonary, bronchial, and renal arteries showed markedly hyperplastic medial elastic laminae approximately three times thick compared to those of age-matched normal artery. The elastic fibers of the innermost two thirds of media were disposed in a normal orderly parallel fashion. In outer third of the media, the elastic fibers had lost the normal orderly arrangement.

Combined Mucinous Tumor and Carcinoid of Appendix Associated with Mucinous Tumor of Ovary and Pseudomyxoma Peritonei: A case report .: Hye Jeong Choi, Mi Jin Kim; Korean J Pathol. 1999;33(11):1094-1096.

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Abstract PDF: We report a case of mucinous cystadenoma of uncertain malignant potential and carcinoid of appendix associated with bilateral mucinous cystadenoma of ovary and pseudomyxoma peritonei. The patient was a 46-year-old female. She suffered from dyspnea and lower abdominal palpable masses for several months. Ultrasonogram showed multilocular huge ovarian cysts. Appendectomy, transabdominal hysterectomy, bilateral salphingo-oophorectomy and biopsy of omentum were performed. The bilateral ovaries measured 16 11 cm and 7X5 cm in size, respectively. The both ovaries showed multilocular cysts filled with thick mucus material. The ovarian cysts were covered by a single layer of columnar epithelium with focal proliferation. Mucus materials dissected through the ovarian stroma (pseudomyxoma ovarii). The tip of appendix was dilated and covered by mucus material. The cut surface showed a cyst and a yellowish solid mass proximal to the cyst. Microscopically, the appendiceal cyst was lined by stratified columnar epithelium with moderate cytologic atypia. Mucus material dissected through the wall. In the proximal portion of the appendix, a classic carcinoid with focal tubular form was present in submucosa and muscle layer. The omentum was covered by thick mucus material. Microscopically, the omentum showed mucinous epithelium and mucus material (pseudomyxoma peritonei).

Multiple Kaposi's Sarcoma in the Renal Transplant Patient: A case report .: Jae Kyung Koh, Eun Sun Jung, Youn Soo Lee, Seok Jin Kang, Byung Kee Kim, Sun Moo Kim; Korean J Pathol. 1999;33(11):1097-1101.

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Abstract PDF: The Kaposi's sarcoma, which was found in an immunosuppressed patient of renal transplantation, may have been developed by long term use of immunosuppressant agent after the renal transplantation. The case was a 29-year-old woman who was diagnosed as chronic renal failure in 1988, and since then, she had been on CAPD until May, 1997. After the renal transplantation in May 1997, the patient has been prescribed cyclosporin and prednisone as immunosuppressant agent. In June 1997, she showed clinical symptom of Kaposi's sarcoma with multiple papules and nodules in the skin and viscera, such as ureter, urinary bladder, stomach, duodenum and subcutaneous tissue of the chest. Multiple excisional biopsies were carried out in the skin, ureter, urinary bladder, stomach and duodenum. All of excisional biopses indicated nodular stages with extensive proliferation of spindle shaped, somewhat pleomorphic cells which have slit-like vascular spaces, proliferation of small vessels, and extravasation of erythrocytes. These lesions nearly diminished after sytemic chemotherpy, excision and discontinuity of immunosuppressive agents.

Polyvesicular Vitelline Tumor of the Ovary: A case report .: Woo Sung Moon, Dong Geun Lee; Korean J Pathol. 1999;33(11):1102-1106.

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Abstract PDF: Polyvesicular vitelline tumor of the ovary is an extremely rare variant of yolk sac tumor. We present a case of pure polyvesicular vitelline tumor in a 43-year-old woman. Light microscopy revealed a predominantly polyvesicular pattern embedded in mesoblastic stroma with the cysts showing two type of lining; tall columnar and cuboidal, or mesothelioid cells. The lining atypical cells showed occasional mitoses and intracytoplasmic PAS positive hyaline globules. In some areas, the cystic space contained a large amount of intraluminal hyaline material. Immunohistochemically, alpha- fetoprotein and alpha-1-antitrypsin were detected as a fine granular deposit in the cytoplasm of epithelial cells and hyaline globules. Electron microscopically, marked specialization of the vesicular lining cells suggested a differentiation toward gut structures and mature yolk sac.

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