• Title/Summary/Keyword: Paraffin-embedded tissue

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Possible Epithelial Ovarian Cancer Association with HPV18 or HPV33 Infection

  • Zhang, Pei-Pei;Zhou, Lei;Cao, Jia-Shi;Li, Yi-Ping;Zeng, Zhi;Sun, Ni;Shen, Li;Zhu, Hao-Yue;Ruan, Yang;Zha, Wen-Ting;Wang, Xin-Yu;Zhang, Ke-Qiang;Zhang, Ran
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.6
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    • pp.2959-2964
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    • 2016
  • The present study was conducted to investigate the prevalence of HPV infection in epithelial ovarian cancer (EOC) in Hunan province. DNA samples were collected from paraffin embedded ovarian tissue from 322 patients with EOC, 99 with ovarian benign tumors and 199 normal persons. The polymerase chain reaction and direct sequencing were used to identify the HPV types in the samples. The relationship between the infection of human papillomavirus (HPV) and the epithelial ovarian carcinoma (EOC) was investigated combined with clinical data. The prevalence of HPV18 and HPV33 in EOC group and benign group was higher than in the normal group. HPV18 and HPV33 may play a role in the development of both EOC and ovarian benign tumor and may participate in the development of EOC with traditional risk factors, family history and abortion, possibly exerting synergistic effects.

Performance of Quantitative Real-Time PCR for Detection of Tuberculosis in Granulomatous Lymphadenitis Using Formalin-Fixed Paraffin-Embedded Tissue

  • Munkhdelger, Jijgee;Mia-Jan, Khalilullah;Lee, Dongsup;Park, Sangjung;Kim, Sunghyun;Choi, Yeonim;Wang, Hye-Young;Jeon, Bo-Young;Lee, Hyeyoung;Park, Kwang Hwa
    • Biomedical Science Letters
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    • v.19 no.2
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    • pp.153-157
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    • 2013
  • Although culture is the gold standard method to identify mycobacteria, its use in tuberculous lymphadenitis (TBL) is limited due to formalin fixation of the submitted specimens. We evaluated the performance of quantitative real-time PCR (q-PCR) for Mycobacterium Tuberculosis (MTB) in granulomatous lymphadenitis using formalin-fixed paraffin-embedded (FFPE) tissues. From 2000 to 2010, a total number of 117 cases of lymph node samples with granulomatous inflammation which were surgically removed and fixed in formalin were studied. Hematoxylin & Eosin (H&E) and Ziehl-Neelsen-stained (ZN) slides were reviewed. qPCR using Real TB-Taq$^{(R)}$ was performed for all cases to identify Mycobacterium tuberculosis. Thirteen non-tuberculous lymphadenopathy cases were used as negative control. Cervical lymph nodes were more frequently affected (60%, 70/117) than other sites. ZN stain for acid fast bacilli was positive in 19 (16.24%) cases. qPCR for tuberculosis was positive in 92 (78.63%) cases. Caseous necrosis was found in 103 (88.03%) cases. While the ZN stain and qPCR were both negative in all control cases, the qPCR showed a significantly higher positive rate (78.63% vs. 16.24%) compared to ZN stain in histologically diagnosed TBL. Quantitative real-time PCR proves to be more sensitive than ZN stain for diagnosis of tuberculous lymphadenitis.

Comorbid Gastric Adenocarcinoma and Gastric and Duodenal Strongyloides stercoralis Infection: A Case Report

  • Seo, An Na;Goo, Youn-Kyoung;Chung, Dong-Il;Hong, Yeonchul;Kwon, Ohkyoung;Bae, Han-Ik
    • Parasites, Hosts and Diseases
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    • v.53 no.1
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    • pp.95-99
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    • 2015
  • Strongyloides stercoralis can cause systemic infection, termed strongyloidiasis, and gastrointestinal ulcer disease in immunocompromised patients. However, to our knowledge, there are no reported cases of comorbid gastric adenocarcinoma and S. stercoralis infection. Here, we report a case of an 81-year-old Korean man who presented with S. stercoralis infection coexisting with early gastric adenocarcinoma (T1aN0M0). S. stercoralis eggs, rhabditiform larvae, and adult females were observed in normal gastric and duodenal crypts. They were also observed in atypical glands representative of adenocarcinoma and adenoma. Preliminary laboratory tests revealed mild neutrophilic and eosinophilic leukocytosis. A routine stool test failed to detect rhabditiform larvae in the patient's fecal sample; however, S. stercoralis was identified by PCR amplification and 18S rRNA sequencing using genomic DNA extracted from formalin-fixed paraffin-embedded tissues. Postoperatively, the patient had a persistent fever and was treated with albendazole for 7 days, which alleviated the fever. The patient was followed-up by monitoring and laboratory testing for 4 months postoperatively, and no abnormalities were observed thus far. The fact that S. stercoralis infection may be fatal in immunocompromised patients should be kept in mind when assessing high-risk patients.

Detection and Characterization of Enteroviral RNA in Paraffin-embedded Heart Tissues from Patients with Dilated Cardiomyopathy

  • Chung, Kyung-Won;Nam, Jung-Hyun;Lee, Ho-Jung;Hong, Hae-Nam;Cho, Young-Keol;Chu, Chul-Hyun;Kim, Yoo-Kyum
    • The Journal of Korean Society of Virology
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    • v.30 no.1
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    • pp.29-37
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    • 2000
  • The aim of this study was to investigate viral etiology in dilated cardiomyopathy (DCM) by polymerase chain reaction (PCR) or nested reverse transcription PCR (RT-PCR), and characterize the enteroviral RNA presented in the clinical specimens. Twenty-eight paraffin-embedded heart tissue samples were assayed to detect cytomegalovirus, herpes simplex virus type 1, type 2, parvovirus, adenovirus, and enterovirus (EV) with each specific primer. Of these 28 patients (mean age: 27, M: 24, F: 4), 26 were histologically diagnosed as DCM and 2 as myocardial infarction (MI). Nested RT-PCR detected enteroviral RNA in 7 (26.9%) of 26 patients with DCM, and none of patients with MI. And none of DNA viruses tested were detected from the samples. Amplified products were also genotyped by single-strand conformation polymorphism (SSCP). Three subtypes can be differentiated from 7 clinical specimens. Furthermore, direct sequence analysis was performed to determine whether genetic variation of EV is present in the explanted heart tissues from patients with DCM. Although most of the sequences among the wild isolates have the greatest similarity to those of coxsackievirus B3, there are specific regions of variable sequences (no 490 - no 510). The data suggest that enterovirus may be a major viral pathogen for the DCM in Korea and nucleotide sequence data indicate that coxsackievirus B3 may be a leading etiologic agent of DCM.

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Detection of porcine encephalomyocarditis virus by in situ hybridization (In situ hybridization에 의한 돼지 뇌심근염 바이러스의 검출)

  • Oh, Sang-hyeon;Park, Nam-yong;Chung, Ci-young;Cho, Kyoung-oh;Lee, Bong-joo;Park, Young-seok;Park, Hyung-seon
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.148-158
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    • 1999
  • The purpose of this study was to establish a rapid, reliable diagnostic method detecting Encephalomyocarditis virus(EMCV) RNA in formalin-fixed, paraffin-embedded tissues of EMCV naturally infected pigs by cDNA probe of EMC $K_3$, the EMCV strain isolated from Korea. Using a biotin-labelled nick translated probe for the cDNA marker. We made up for some defects of radiolabeled method. In sits hybridization(ISH) technique, differently from the other nucleic acid hybridization methods, is able to detect the virus genome specifically in the state of the intact shapes of cells and/or tissues. We succeeded in performing the experiment to detect the EMCV within 1~2 hours using the $MicroProbe^{TM}$ capaillary action system. In this study, we observed highly specific positive signals of red color by staining the paraffin-embedded tissue sections of naturally EMCV-infected pig organs or tissues, including brain, heart, kidney and lacrimal gland with the Fast Red TR salt/Naphtol phosphate chromogen. The results suggested that this ISH method is considered as a highly sensitive and reliable tool for molecular biologic diagnosis of the EMC viral disease.

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Construction of Artificial Epithelial Tissues Prepared from Human Normal Fibroblasts and C9 Cervical Epithelial Cancer Cells Carrying Human Papillomavirus Type 18 Genes

  • Eun Kyung Yang;Seu
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.1
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    • pp.1-5
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    • 1998
  • One cervical cancer cell line, C9, carrying human papillomavirus type 18 (HPV18) genes that is one of the major etiologic concoviruses for cervical cancer was characterized. This cell line was further characterized for its capacity related to the epithelial cell proliferation, stratification and differentiation in reconstituted artificial epithelial tissue. The in vitro construction of three dimensional artificial cervical opithelial tissue has been engineered using C9 epithelial cancer cells, human foreskin fibroblasts and a matrix made of type I collagen by organotypic culture of epithelial cells. The morphology of paraffin embedded artificial tissue was examined by histochemical staining. The artificial epithelial tissues were well developed having multilayer. However, the tissue morphology was similar to the cervical tissus having displasia induced by HPV infection. The characteristics of the artificial tissues were examined by determinining the expression of specific marker proteins. In the C9 derived artificial tissues, the expression of EGF receptor, as epithelial proliferation marker proteins for stratum basale was observed up to the stratum spinosum. Another epithelial proliferation marker for stratum spinosum, cytokerations 5/6/18, were observed well over the stratum spinosum. For the differentiation markers, the expression of involucrin and filaggrin were observed while the terminal differentiation marker, cytokeratins 10/13 was not detected at all. Therefore the reconstituted artificial epithelial tissues expressed the same types of differentiation marker proteins that are expressed in normal human cervical epithelial tissues but lacked the final differentiation capacity representing characteristics of C9 cell line as a cancer tissue devived cell line. Expression of HPV18 E6 oncoprotein was also observed in this artifical cervical opithelial tissue though the intensity of the staining was weak. Thus this artificial epithelial tissue could be used as a useful model system to examine the relationship between HPV-induced cervical oncogenesis and epithelial cell differentiation.

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AN IMMUNOHISTOCHEMICAL STUDY ON THE CELLULAR CHANGE IN EPITHELIUM AND SUBEPITHELIAL TISSUE OF NON-INFLAMMATORY GINGIVAL HYPERPLASIA (비염증성 치은증식증의 상피 및 상피하조직내 세포변화에 관한 면역조직화학적 연구)

  • Choi, Yeoung-Wook;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.23 no.3
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    • pp.605-621
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    • 1993
  • The gingival hyperplasia refers to an increase in the size of the gingival tissue produced by an increase in the number of its component cells. In order to investigate the cellular change in epithelium and subepithelial tissue of noninflammatory gingival hyperplasia, the gingival tissues were surgically obtained from the patients with dilantin gingival hyperplasia and idiopathic gingival hyperplasia. The excised tissue samples were fixed in neutral formalin for 6-24 hours, embedded with paraffin, sectioned at $4-6{\mu}m$ in thickness, mounted on glass slides coated with 3-aminopropyltriethoxysilane(Sigma Chemical Co., St. Louis, MO, U.S.A.) and immunocytochemically processed by Avidin-Biotin peroxidase complex method for detecting proliferating cell nuclear antigen, tenascin and collagen type IV. Monoclonal mouse anti-human PCNA antibody(Oncogene Science, Uniondale, NY, U.S.A., 1 : 250,000), monoclonal mouse anti-human tenascin antibody(Chemicon-International Inc., Temecula, CA, U.S.A., 1:5,000), and monoclonal mouse anti-human collagen type IV(Dakopatts, Glostrup, Denmark, 1: 50) were used as primary antibodies. The results were as follows: 1. In non-inflammatory gingival hyperplasia, the positive reaction to proliferating cell nuclear antigen was localized in the basal cell layer of gingival epithelium and well-developed rete pegs. 2. The positive reaction to tenascin was shown in the connective tissue subjacent to basament membrane of gingival tissue, and especially strong positive reaction was noted in the tip portion of connective tissue projections. 3. The positive reaction to collagen type IV was localized along the basement membranes of gingival epithelium and blood vessels. The results suggest that connective tissue enlargement may affect the proliferation of gingival epithelium.

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AN IMMUNOHISTOCHEMICAL STUDY OF THE DISTRIBUTION OF FIBRONECTIN, LAMININ AND TENASCIN IN THE REGENERATING PERIODONTAL TISSUE (재생중인 치주조직내 Fibronectin, Laminin 및 Tensacin의 분포에 관한 면역조직화학적 연구)

  • Chung, Gap-Hwan;Kim, Byung-Ok;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.321-340
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    • 1995
  • The regeneration of destructed periodontal tissues is one of the ultimate objectives of periodontal therapy. Guided tissue regeneration technique was developed for the ideal regeneration of periodontal tissues. In order to investigate the role of fibronectin, laminin and tenascin in the regenerating process of periodontal tissues, the expanded PTFE barrier membranes(Gore Associates, USA) removed from the patients who had been treated by guided tissue regeneration(GTR) and guided bone regeneration(GBR) techniques were fixed in neutral formalin for 6-24 hours, embedded with paraffin, sectioned at $4-6{\mu}m$ in thickness, and immunohistochemically processed by Avidin-Biotin peroxidase complex method for detecting fibronectin, laminin and tenascin. Monoclonal mouse anti-human fibronectin antibody(Oncogene Science, USA., 1:100), monoclonal mouse anti-human laminin antibody(Oncogene Science, USA., 1:50) and mouse anti-human tenascin antibody(Oncogene Science, USA, 1:10) were used as primary antibodies. The light microscopic findings were as follows: (1) The distribution of fibronectin, laminin and tenascin was various according to the area of barrier membranes. (2) The distribution of fibronectin in case of GBR was extensive in the tissue on the outer surface of barrier membranes, and rare in the intervening space and on the inner surface. In case of GTR it was extensive on the outer surface and in the intervening space, and rare on the inner surface. (3) The distribution of laminin was rare in the tissue on the outer, the inner surface and intervening space of barrier membranes, regardless of GBR or GTR. (4) In case 'of GBR rare distribution of tenascin was observed on the outer surface only, except the inner surface and the intervening space of barrier membranes. In case of GTR the distribution of tenascin was extensive in the tissue on the outer surface, rare in intervening space and the inner surface. The results suggest that fibronectin, laminin and tenascin may play a important role in the regenerating process of periodontal tissue, and they may affect the outcome of healing.

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AN EXPERIMENTAL STUDY ON THE EFFECT OF CONDYLAR OSTEOPLASTY WITH PRESERVATION OF ARTICULAR COVERED SOFT TISSUE ON THE HEALING PROCESS IN RABBIT (가토 하악과두 연조직 피개가 과두골성형술시 치유과정에 미치는 영향에 관한 실험적 연구)

  • Jang, Dong-Ho;Lee, Dong-Keun;Kim, Soo-Nam
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.13 no.3
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    • pp.241-251
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    • 1991
  • During the condylar shaving procedure, the articular soft tissue cover can be removed. Author compaired the histological healing process of the articular soft tissue cover between the preservative and unpreservative group group with 45 New Zealand rabbits(Average wt. : about 2.5kg). In unpreservative group, the usual high condylar shave with the removal of soft tissue cover was performed. In the preservative group, the underlying bone, replaced in its original position and sutured. The animals were sacrified 1, 2, 3, 4, 6 weeks interval after operation. The specimens were fixed in 10% neutral formalin and decalcified, paraffin embedded and stained by Hematoxylin & Eosin, and Masson's trichrome. The obtained results were as follows. 1. The condyles of the both group were covered with an articular sop tissue layer. 2. The cartilage cells in subarticular layer has regular continuous patterns in the preservative group but frequently interrupted in the unpreservative group. 3. The incision made in the posterior part of the articular surface for the elevation of the articular soft tissue frequently caused a deformity such as the interruption of the subarticular layer of cartilage. 4. By the above findings, the preservation of articular sop tissue cover may be the effective operation method on concept of bone remodelling.

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Feline infectious peritonitis : A case report (고양이 전염성복막염의 자연감염 예)

  • ;;;;;;Manfred Reinacher
    • Korean Journal of Veterinary Service
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    • v.26 no.1
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    • pp.67-71
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    • 2003
  • A 8 months old, female domestic Shorthair cat with long-term signalment of anorexia, lacrimation, uveitis and coughing was submitted to the Pathology and Diagnosis Reference Division, NVRQS, Korea, for necropsy. Main gross lesions were characterized by ascities, some grayish-white nodular formation and fibrous adhesion on the surface of visceral organs including liver and kidney. Principle histopathological findings were fibrinous serositis, multifocal granuloma and necrosis, vasculitis, perivasculitis in various pharenchymal organs. Paraffin-embedded tissue sections taken from most of organs with granulomatous lesions were confirmed specific reaction to the monoclonal antibody of feline infectious peritonitis virus in the cytoplasm of many infiltrating macrophages by immunohistochemistry. The report was to describe the pathological lesions of the first naturally-occuring FIP case in companion cat of Korea.