• Title/Summary/Keyword: paraffin-embedded tissue

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Expression of Bcl-2 in the epithelial lining and clinical findings of keratocystic odotogenic tumor (각화낭치성종양의 이장상피에서 Bcl-2 발현양상과 임상지표)

  • Lee, In-Hyuk;Choi, So-Young;Park, Ji-Hoon;Kim, Chin-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.3
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    • pp.161-168
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    • 2011
  • Purpose: The odontogenic keratocysts demonstrated a high recurrence rate and a biologically aggressive nature. This might be due to unknown factors inherent in the epithelium or enzymatic activity in the fibrous wall. Bcl-2 protein is characterized by its ability to inhibit apoptosis. The aim of this study was to evaluate the expression and distribution of bcl-2 in the OKCs, its possible relationship with the tumorous characteristics, such as the aggressive nature and high recurrence rate, and its usefulness to differentiate OKCs from dentigerous cysts. Materials and Methods: Formalin fixed paraffin-embedded tissue sections of 53 OKCs, and 44 dentigerous cyst were immunohistochemically analyzed quantitatively for the immunoreactivity of the bcl-2 protein with i-solution. Results: More Bcl-2 expression was observed in the OKCs (mean34.387%) than dentigerous cyst (mean11.144%) with statistical significance (P<0.001). Seventeen and 15 of the 32 OKCs in this study showed positivity in the basal layer and basal/suprabasal layers, respectively. In dentigerous cyst, 2 of 3 showed positivity in the basal cell layer. Conclusion: Considering that bcl-2 over expression may lead to the increased survival of epithelial cells, this study demonstrated a possible relationship between the aggressive nature of OKC and the intrinsic growth potential of its lining epithelium. Furthermore, the basal/suprabasal distribution of bcl-2 positive cells was observed in some OKCs, which might have a significant impact on the behavior of cysts. The bcl-2 expression of OKCs can be useful for differentiating OKCs from dentigerous cysts.

Detection of Human papillomavirus & Ebstein-Barr virus in Sinonasal Inverted papilloma (비강 및 부비동의 반전성 유두종에서 인형유두종 및 Ebstein-Barr 바이러스의 검출)

  • Cho, Jae-Shik;Lim, Sang-Chul;Baik, Jun;Jeong, Hyoung-Soo;Shin, Myoung-Seok
    • Korean Journal of Bronchoesophagology
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    • v.5 no.1
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    • pp.49-54
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    • 1999
  • The etiology of inverted papilloma(IP) remains unknown, but several studies have reported that Human Papillomavirus(HPV) may play a role in the pathogensis of sinonasal inverted papilloma(IP). And recent reports demonstrate the possible etiologic role of Epstein-Barr virus (EBV) in sinonasal IP. The aim of this study is to detect HPV and EBV in sinonasal IP, to examine the relationship between HPV subtype and sinonasal IP, to investigate the relation between HPV and EBV. We reviewed 30 cases of sinonasal IP(simple IP 19 cases, IP with dysplasia 8 cases, IP with squamous cell carcinoma 3 cases). Paraffin embedded archival tissue was used in this study. Detection of HPV, EBV were examined by in situ hybridization(ISH) using HPV type 6/11, 16/18, 31/33/35 DNA probe and EBER probe. The HPV was detected in 6(20%) out of 30 cases. The HPV 6/11 was dectected in 4 out of 19 cases of simple IP, HPV 16/18 in 1, HPV 31/33/35 in 1 out of 8 cases of IP with dysplasia respectively. The EBV was not detected in 30 cases. HPV may play a role in the pathogensis of sinonasal inverted papilloma. But EBV is not a etiopathologic factor to be considered in the development of sinonasal IP.

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EXPRESSION OF CYTOKERATIN SUBTYPES AND VIMENTIN IN AMELOBLASTOMA (사기질모세포종에서 Cytokeratin 아형과 Vimentin의 발현)

  • Kang, Mi-Seon;Yoon, Hye-Kyoung;Kim, Woo-Hyung;Choi, Soo-Im
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.4
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    • pp.316-321
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    • 2005
  • Ameloblastoma is the most common odontogenic tumor of the jawbones, but the origin of this tumor has been remained to be unproven. Cytokeratins (CKs) are specific intermediate filament of epithelial cells, and vimentin is expressed in mesenchymal cells. The immunohistochemical detection of different CKs and vimentin has made it easier to know the origin of tumor. Paraffin-embedded tissue sections from 15 ameloblastomas and 1 ameloblastic carcinoma were used for immunohistochemical evaluation of CK 7, 8, 13, 14, 19 and vimentin. Their expression is evaluated in different tumor cells, which are observed in different type of tumors. In the follicular and reticular subtype, central stellate cells of tumor nests expressed CK 8, 14, 19 and peripheral columnar cells expressed CK 14. CK 7, and 13 were not expressed. Vimentin was detected in fibrous stroma around tumor nest, not in tumor cells. The tumor cells of ameloblastic carcinoma expressed CK 7, 14 and 19, but CK 8 was more weakly stained than that in ameloblastoma. Central stellate cells and peripheral columnar cells of acanthomatous subtype showed same expression pattern with others. Meta plastic squamous cells expressed CK 8, 14, 19 and keratinizing squamous cells expressed CK 13, 19. CK 7 and vimentin were not detected in tumor cells and vimentin was expressed in fibrous stroma. Most of the tumor cells of ameloblastoma showed CK 14 and CK 19 and did not express CK 7 and vimentin. These findings were similar to the immunophenotype of dental lamina. And these results will be beneficial to differential diagnosis of odontogenic tumors and other kind of tumors arising at the oral cavity.

AN EXPERIMENTAL STUDY ON THE CHANGE OF CONDYLE HEAD AFTER MANDIBULAR RAMUS OBLIQUE OSTEOTOMY (하악지 사선골절단술 후 하악두의 변화에 관한 실험적 연구)

  • Cha, Seon-Kyung;Kim, Yeo-Gab
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.14 no.1_2
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    • pp.65-76
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    • 1992
  • This study was designed to observe the adaptive changes of mandibular condyles to displacement of mandibular condyle in adult animals. In this study, 16 rabbits weighting about 3.5 kg was selected. Four rabbits were preserved for control group and 8 animals were divided into 3 groups, 2 weeks, 4 weeks and 8 weeks. The experimental animals were performed oblique osteotomy on both mandibular ramus and internal wiring at mandibular border. The experimental animals were sacrificed consecutively on the 2 weeks, 4 weeks and 8 weeks after oblique osteotomy and mandibular condyles were dissected out carefully to produced tissue specimen. The specimens were fixed with 10% N formaline solution for 24 hours and rinsed with phosphate buffer solution. It was decalcified with 5% nitric acid for 15 days. Thereafter the specimens were dehydrated in alcohol series and embedded paraffin as usual manner. The mebedded specimen were sectioned in $4-6{\mu}m$ microtome, stained with hematoxylin-eosin and azan stain and observed through light microscope. The following results were observed from this experiment. When there was postional change of condyle head after mandibular ramus oblique osteotomy in adult rabbit, 1. The density of chondrocyte was generally increased at condylar cartilage and the thickness of condylar cartilage was increased posterosuperior aspect of the mandibular condyle slightly. 2. The density of chondrocyte was increased at proliferative zone so fibrous articular zone, porliferative zone and hypertrophic zone was clearly distinguished. 3. Active endochondral bone formation was occurred at mandibular condyle.

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THE EFFECTS OF ODONTOGENIC AND NONODONTOGENIC TISSUES ON BONE HEALING IN GUINEA PIG MANDIBLE (치성 및 비치성 조직이 악골 창상치유에 미치는 영향에 관한 실험적 연구)

  • Kim So-Jung;Hwang Eui-Hwan;Lee Sang-Rae;Hong Jung-Pyo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.26 no.2
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    • pp.33-44
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    • 1996
  • This study was for comparing healing patterns and effects between with odontogenic and nonodontogenic tissues on the defected mandible. Experimental bone defects that measured 3 mm in diameter were created on the mandibular body of guinea pig by removal of bone with the use of trephine burs and bone defects were grafted with Biogran(Orthovita Co., U.S.A.) and covered with Dura Mata(Pfrimmer-Viggo GmbH Co., Germany). Guinea pigs were serially terminated by fours on the 3 days, the 1 week, the 2 weeks, the 3 weeks, the 4 weeks, and the 5 weeks after experiment, and the mandibular body was removed and fixed with 10% neutral formalin. They were decalcified and embedded in paraffin as using the usual methods. The specimen sectioned and stained with hematoxylin and eosin and toluidine blue. They were observed with a light microscope and a polarizing microscope. The obtained results were as follows : 1. Defected bone was healed fast from the odontogenic tissues in early stage of the experiment. 2. The arrangement of the bone matrix was relatively regular in the bone from the nonodontogenic tissues, but irregular in the bone from the odontogenic tissues. 3. Compact bone has started to be resorbed and changed to the pattern of matrix bone tissue from 3 weeks after experiment.

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Studies on the pathogenesis of Korean isolate of Aujeszky's disease virus in experimentally infected piglets II. Immunohistochemistry and detection of viral nucleic acids by in situ hybridization (Aujeszky's disease virus 국내분리주 접종자돈의 병리발생에 관한 연구 II. 면역조직화학 및 in situ hybridization 기법을 이용한 항원과 핵산 검출)

  • Cho, Woo-young;Cho, Sung-whan;Park, Choi-gui;Kim, Jae-hoon;Hyun, Bang-hoon;Yoon, Yong-dhuk;Kweon, Chang-hee
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.859-871
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    • 1996
  • This study was conducted to elucidate the distribution of Aujeszky's disease viral nucleic acids and antigens in the central nervous system (CNS) of piglets. The first Korean isolate of Aujeszky's disease virus(ADV) that isolated from naturally infected piglets in Yang San, was inoculated into 32 day old piglets with $10^{5.9}TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at every 24hrs for 8 days. The immunohistochemistry (IHC) was conducted to detect the viral antigens in paraffin-embedded tissue sections using avidin-biotin-peroxidase complex (ABC) method. The viral nucleic acids were detected by in situ hybridization (ISH) using ADV specific DNA probe labeled with digoxigenin. The ADV antigens were detected in reticuloendothelial cells of spleen, lymph nodes and tonsil, alveolar walls, leptomeningeal vascular walls, inflammatory foci of each organ, and nerve cells. The viral nucleic acids were detected in the spinal trigeminal nucleus and its tracts of the pons and medulla oblongata by the ISH technique. The pathways of AD viruses in CNS were determined by IHC and ISH. In the intranasally inoculated group, the viruses in nasal mucosa moved to medulla oblongata and pons through the trigeminal nerve. In case of intramuscullarly inoculated group, viruses moved to brain via lymphoid organs or spinal nerves from sciatic nerves.

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Objective Quantitation of EGFR Protein Levels using Quantitative Dot Blot Method for the Prognosis of Gastric Cancer Patients

  • Xin, Lei;Tang, Fangrong;Song, Bo;Yang, Maozhou;Zhang, Jiandi
    • Journal of Gastric Cancer
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    • v.21 no.4
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    • pp.335-351
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    • 2021
  • Purpose: An underlying factor for the failure of several clinical trials of anti-epidermal growth factor receptor (EGFR) therapies is the lack of an effective method to identify patients who overexpress EGFR protein. The quantitative dot blot method (QDB) was used to measure EGFR protein levels objectively, absolutely, and quantitatively. Its feasibility was evaluated for the prognosis of overall survival (OS) of patients with gastric cancer. Materials and Methods: Slices of 2×5 ㎛ from formalin-fixed paraffin-embedded gastric cancer specimens were used to extract total tissue lysates for QDB measurement. Absolutely quantitated EGFR protein levels were used for the Kaplan-Meier OS analysis. Results: EGFR protein levels ranged from 0 to 772.6 pmol/g (n=246) for all gastric cancer patients. A poor correlation was observed between quantitated EGFR levels and immunohistochemistry scores with ρ=0.024 and P=0.717 in Spearman's correlation analysis. EGFR was identified as an independent negative prognostic biomarker for gastric cancer patients only through absolute quantitation, with a hazard ratio of 1.92 (95% confidence interval, 1.05-3.53; P=0.034) in multivariate Cox regression OS analysis. A cutoff of 208 pmol/g was proposed to stratify patients with a 3-year survival probability of 44% for patients with EGFR levels above the cutoff versus 68% for those below the cutoff based on Kaplan-Meier OS analysis (log rank test, P=0.002). Conclusions: A QDB-based assay was developed for gastric cancer specimens to measure EGFR protein levels absolutely, quantitatively, and objectively. This assay should facilitate clinical trials aimed at evaluation of anti-EGFR therapies retrospectively and prospectively for gastric cancer.

Comparative Morphological Study on Parotid and Submandibular Salivary Glands in Ovariectomized Rats

  • Jeong, Moon-Jin;Lee, Myoung-Hwa;Lim, Do-Seon;Jeong, Myeongju;Jeong, Soon-Jeong
    • Journal of dental hygiene science
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    • v.22 no.2
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    • pp.83-89
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    • 2022
  • Background: Estrogen deficiency affects the structure and function of the salivary glands in women, leading to a decrease in salivary secretion and a change in the composition of saliva. Previous studies on changes in the salivary glands that cause estrogen deficiency have reported only partial results for the parotid and submandibular glands, and there are few comparative morphological studies of histological changes between the parotid and submandibular glands in ovariectomized rats (OVX) leading to estrogen deficiency. This study aimed to analyze the histopathological and histochemical changes in the parotid and submandibular salivary glands causing estrogen deficiency by using OVX, and to discuss the mechanism on these changes. Methods: The parotid and submandibular glands from sacrificed control and OVX groups were fixed with cold 4% paraformaldehyde in phosphate buffer (pH 7.2). The tissues were dehydrated using a series of graded ethyl alcohol and embedded in paraffin. For histopathological analysis, sections cut to a thickness of 6 to 7 ㎛ were stained with hematoxylin and eosin (H&E). For histochemical analysis, Periodic acid-Schiff (PAS), Alcian blue (AB, pH 2.5), and PAS+AB (pH 2.5 and pH 1) staining was performed. Results: Histopathological analysis of OVX tissue showed that the parotid and submandibular salivary glands were broadly and clearly separated and divided into lobes. In OVX, acinar and ductal cells with condensed polymorphic or pyknotic nucleus, which are presumed to be characteristic of apoptotic cells, and degenerated cells with lipid deposition in cytoplasmic granules and ruptured membranes were increased. Histochemical analysis of OVX, confirmed an increase in the number and acidification of acinar secretory granules. Conclusion: Histopathological and histochemical changes and the effects of estrogen deficiency are more evident in the submandibular salivary gland than in the parotid gland.

Evaluating the Efficacy of a Formalin Alternative in Fixing Pathological Tissues for Histological and Molecular Diagnoses

  • Min-A Je;Haneul Lee;Heechul Park;Dong Hyeok Kim;Yeongdon Ju;Jaewon Lim;Sunghyun Kim;Jungho Kim
    • Biomedical Science Letters
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    • v.29 no.1
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    • pp.48-52
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    • 2023
  • Formaldehyde use is associated with serious health risks, which can affect medical personnel and technicians. Therefore, we investigated the efficacy of an alternative fixative, with respect to two types of formalin fixatives, by hematoxylin and eosin (H&E) staining, periodic acid Schiff (PAS) staining, immunohistochemical (IHC) staining, and RNA extraction. For H&E staining, the circular nucleus was stained dark blue by the basic dye hematoxylin and the cytoplasm was stained red by the acid dye eosin in all three fixative samples. No difference was found in the Duksan General Science (DGS), Sigma-Aldrich, and Core-Fix fixative samples (Corebiotech) used to fix kidney tissue, after PAS staining. IHC staining showed that CD4 was significantly increased in the lippolysaccharide (LPS)-treated group compared to the control group (vehicle), confirming the changes in specific molecules. The quantity and quality of RNA from tissues fixed in the three types of fixatives were evaluated. The average concentration of RNA was 106 ng/µL and average purity at A 260/280 ratio was 1.7~2.0, regardless of fixative used. For quality of protein, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was confirmed by Western blotting. In conclusion, Core-Fix can be used as a fixative for pathological tissues, in histological and molecular diagnoses.

Interplay of collagen and mast cells in periapical granulomas and periapical cysts: a comparative polarizing microscopic and immunohistochemical study

  • Deepty Bansal;Mala Kamboj;Anjali Narwal;Anju Devi;Nisha Marwah
    • Restorative Dentistry and Endodontics
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    • v.47 no.1
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    • pp.12.1-12.11
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    • 2022
  • Objectives: This pilot study aimed to establish the interrelationship between collagen and mast cells in periapical granulomas and periapical cysts. Materials and Methods: An observational cross-sectional study was conducted on the paraffin-embedded tissue sections of 68 specimens (34 periapical granulomas and 34 periapical cysts). The specimens were stained with picrosirius to observe collagen fiber birefringence and anti-tryptase antibody to evaluate the mast cell count immunohistochemically. The mean number and birefringence of collagen fibers, as well as the mean number of mast cells (total, granulated, and degranulated), and the mean inflammatory cell density were calculated. The data obtained were analyzed using the Kruskal Wallis test, Mann Whitney U test, and Spearman correlation test (p < 0.05). Results: The mean number of thick collagen fibers was higher in periapical cysts, while that of thin fibers was higher in granulomas (p = 0.00). Cysts emitted orange-yellow to red birefringence, whereas periapical granulomas had predominantly green fibers (p = 0.00). The mean inflammatory cell density was comparable in all groups (p = 0.129). The number of total, degranulated, and granulated mast cells exhibited significant results (p = 0.00) in both groups. Thick cyst fibers showed significant inverse correlations with inflammation and degranulated mast cells (p = 0.041, 0.04 respectively). Conclusions: Mast cells and inflammatory cells influenced the nature of collagen fiber formation and its birefringence. This finding may assist in the prediction of the nature, pathogenesis, and biological behavior of periapical lesions.