• Title/Summary/Keyword: negative staining

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TISSUE CHANGE AFTER EMBEDDING GELATIN MATRIX IMPLANT(FFIBREL®) IN SUBCUTANEOUS TISSUE OF RATS;HISTOLOGIC, IMMUNOHISTOCHEMICAL AND SCANNING ELECTRON MICROSCOPIC STUDY (백서의 피하조직에 Gelatin Matrix Implant (Fibrel®) 매식시 조직변화에 관한 연구)

  • Kim, Hong-Jin;Lee, Chong-Heon;Kim, Kyung-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.20 no.4
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    • pp.341-354
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    • 1998
  • GMI (Fibrel${(R)}$) is one of the dermal filling substances which have been successfully used for the treatment of depressed cutaneous scar and wrinkles. It's major components are; Gelatin powder, which provides a framework for the clot to form and remains stable under the scar, and ${\varepsilon}$-aminocaproic acid, which inhibits the production of fibrinolysin, and Plasma, which provides the necessary ingredients for collagen synthesis. GMI has advantages of low immunogenicity and increased longevity. It has been known to induce fibroblast activity and promote new collagen synthesis. We used 34 Sprague-Dawley rats which were bred under the same condition and duration. 18 of experimental animals were undergone cardiac puncture, and their blood were collected, centrifugated, and stored in freezer. Out of 16 animals, control group were injected with 2ml plasma into the subcutaneous tissue of Lt. scapular, while experimental group were implanted of 2 ml GMI into the Rt. same area. Experimental animals were sacrificed at the 3rd day, 5th day, 1st week and 2nd week respectively after implantation of GMI. To observe the histopathologic change of GMI and surrounding tissue reaction of GMI, we had examined with H&E staining, immunohistochemical staining with vimentin, ${\alpha}$-SMA, S-100 under LM and SEM. The obtained results were as follows ; 1. In LM study, the inflammatory cell infiltrations and granulation tissue formation were observed, and muscle tissues were well attached with adipose tissues in the control group. In the experimental group, inflammatory cell infiltrations had been observed by the 2nd week and irregular adipiose tissues and well differentiated mesenchymal tissues were examined. 2. In immunohistochemical study, the experimental group of ${\alpha}$-SMA study, there were a prominent positive response on endothelial development of granulation tissues and mesenchymal tissues compare with the control group. In vimentin study, positive response on mescenchymal fibroblast continued to 2nd week, but negative in the control group. In S-100 study, both groups were positively responded on irregular adipose tissues. 3. In SEM study, collagen fibers were embedded by the plasma by the 5th day in the control group, and in the 3rd day experiment GMI were resorved but communited with collagen fiber till the 1st week. Collagen fibers were infilt-rated into GMI at the 2nd week and the infilltrated GMI were conglomerated with the mature adipose cells and the collagen fibers. From the above results, GMI implantation in the subcutaneous tissue of Sprague-Dawley rat, the mild infiltration of inflammatory cells were showed till 2nd week and the granulation tissues were observed. GMI were nearly resorbed till 2nd week, but well attached with adipose tissue and collagen fibers. The endothelium and fibroblasts were actively proliferated. Adipose tissues and mesenchymal tissue cells were observed. As already expressed, GMI showed resorptive change in course of time without any early immune reaction, and seemed to induce fibroblast activity and promote new collagen synthesis.

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Immunohistochemical Diagnosis of Renal Pelvis Transitional Cell Carcinoma in a Dog (개의 신우 이행상피암종의 면역조직화학적 진단)

  • Yang, Hyoung-Seok;Kang, Sang-Chul;Kim, Jae-Hoon
    • Journal of Veterinary Clinics
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    • v.30 no.4
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    • pp.292-295
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    • 2013
  • A 9-year-old, male, Doberman pinscher dog with 5-month history of intermittent hematuria, vomiting and glucosuria was referred to local animal hospital. Abdominal ultrasonography showed an irregular and hyperechoic mass in the renal medulla of the enlarged left kidney. Grossly atrophied renal cortex and medulla and marked hydronephrosis were observed on the cut surface of kidney. A single, numerous papillary projected, pedunculated mass 4~5.5 cm in diameter was occupied in renal pelvis, and extended from pelvis to the inlet of ureter. Histopathologically, the mass had numerous papillary structures with arboriform pattern. These papillae were consisted of fibro-vascular stalks that were lined by multiple layers of neoplastic urothelium (transitional epithelium) with marked cellular atypia. Immnohistochemical (IHC) staining demonstrated that the neoplastic cells showed strong positive reactions for cytokeratin (CK) 7, CK 19, CK clone MNF116 and CK high molecular weight, but negative signals for CK 8 low molecular weight. Based on the gross findings, histopathology and CKs profile using IHC staining, this mass was diagnosed as renal pelvis transitional cell carcinoma in a dog.

PROGNOSTIC VALUE OF PROLIFERATION FACTORS IN ORAL SQUAMOUS CELL CARCINOMA (구강 편평세포암에서 예후 및 경부임파절 전이와 관련한 증식인자의 발현에 관한 연구)

  • Paeng, Jun-Young;Ok, Yong-Ju;Yun, Pil-Young;Myoung, Hoon;Hwang, Soon-Jung;Seo, Byoung-Moo;Choi, Jin-Young;Lee, Jong-Ho;Choung, Pill-Hoon;Kim, Myung-Jin
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.2
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    • pp.101-106
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    • 2006
  • Purpose: The purpose of this study is to evaluate the value of proliferation factors, Ki67 and PCNA, as prognostic markers predicting the survival and neck metastasis in patients with oral cancer. Methods: 101 patients with HNSCCs, were followed retrospectively for a median period of 60 months(from 16 to 82 months). All tumors were resected surgically and examined by conventional light microscopy, immunohistochemistry. The age, sex, tumor location, clinical stage(size), metastasis, proliferative activity index(assessed by proliferating cell nuclear antigen(PCNA) and Ki67 immunoreactivity) were considered as potential prognostic factors and were correlated with patient survival. Results: Ki67 staining results ranged from 5% to 80% of tumor cell nuclei, with a median of 25%. PCNA staining results ranged from 1% to 90% with a median of 50%. With a cut-off point of 25%, patients with lower Ki67 scores showed survival advantages over those with higher Ki67 scores (p=0.0089). With cut-off point of 50%, patients with lower PCNA scores showed survival advantages over those with higher PCNA scores (p=0.0104). Pathologically neck node positive patients(n=27) showed higher PCNA expression(p=0.02) than pathologically negative neck node patients(n=39). Conclusions: The lower expressions of Ki67 and PCNA were associated with favorable prognosis such as higher survival rate and lower neck node metastasis.

Correlation between Infiltrations of Tumor-associated Macrophages, Mast Cells, and Dendritic Cells with Clinicopathologic Factors in Advanced Gastric Cancer (진행성 위암에서 종양 연관성 대식세포, 비만세포, 가지세포의 침윤과 임상-병리학적 인자와의 연관성)

  • Lee, Seung-Bum;Chi, Kyong-Chon
    • Journal of Gastric Cancer
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    • v.5 no.3 s.19
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    • pp.206-212
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    • 2005
  • Purpose: Angiogenesis has a critical role in tumor proliferation, invasion, and metastasis. In gastric cancer, tumor-associated macrophages and mast cells produce angiogenic factors such as VEGF, that inhibit the functional maturation of dendritic cells. The aim of this study is to identify tumor-associated macrophages, mast cells, dendritic cell infiltrations, and microvessel densities (MVD) to investigate the relationship between them and the prognosis for gastric-cancer patients. Materials and Methods: The subjects were 79 patients selected from those who had undergone a curative gastric resection for stomach cancer. With them, Immune-histochemical staining was done using CD34 for the MVD, CD68 antigen for macrophages, and S-100 protein for dendritic cells, and toluidine blue staining was done for mast cells. Results: Macrophage infiltration showed a statistically significant positive correlation with histologic differentiation and a negative correlation with invasion depth, nodal metastasis, and stage. S-100 (+) dendritic cells and mast cells had no significant correlations with histologic differentiation, invasion depth, nodal metastasis, distant metastasis, stage, and MVD. As survival, no statistically significant differences were seen between the variables. Conclusion: Tumor-associated macrophages should be evaluated as possible prognostic markers in gastric-cancer patients.

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Correlations of Oral Tongue Cancer Invasion with Matrix Metalloproteinases(MMPs) and Vascular Endothelial Growth Factor(VEGF) Expression (Matrix Metalloproteinases(MMPs) 및 Vascular Endothelial Growth Factor(VEGF)의 발현을 통한 구강 설암의 침윤 기전 연구)

  • Kim Se-Heon;Cho Nam-Hoon;Lim Jae-Yul;Kim Ji-Hoon;Kim Jeong-Hong;Chang Jung-Hyun;Choi Eun-Chang
    • Korean Journal of Head & Neck Oncology
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    • v.21 no.1
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    • pp.3-9
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    • 2005
  • Purpose: In oral tongue cancer, the degree of tumor invasion has a significant effect on the prognosis. We hypothesized that the destruction of extracelluar matrix and neovascularization are related to tumor infiltration mechanism. By studying the the tissues of early stage oral tongue cancer patients, we are intend to clarify the invasion related factors in oral tongue cancer. Material and Methods: To demonstrate the invasion process in early T-stage oral tongue cancer, the expressions of extracellular matrix destruction related molecules(MMP2, MMP9) and neovascularization related molecule(VEGF) were observed by immunohistochemical study. Also, immunohistochemical staining of CD31 was done for quantification of neovascularization. With the experiment showed above, we analyzed relationship between expression of each substances and tumor invasion depth, tumor free survival rates and cervical lymph node metastasis rate in early T-stage oral tongue cancer. Results: The expression rates of MMP2, MMP9, VEGF in 38 early oral cancer patients were 52.6%, 78.9% 52.6%, respectively. Significant correlation was found between the VEGF expression and microvessel density showed by CD31 immunohistochemical staining(p<0.001). VEGF expressions were significantly related with tumor invasion depth(p=0.002). The tumor free survival rate of those patients with VEGF-positive tumors was significantly poorer than in those with VEGF-negative tumors(p=0.019). Conclusion: This results indicate that VEGF is a useful marker for predicting the tumor invasion in patients with early tongue cancer and could be used as a beneficial factors in defining operative field and prognosis.

The Efficacy of β-cyclodextrin/polyethyleneimine/silk Fibroin Hydrogel in Healing Burnt Wound (사이클로덱스트린/폴리에틸렌이민/실크 피브로인 수화겔의 화상치유효능)

  • Seo, Seung Ree;Lee, Mi Sun;Kim, Jin-Chul
    • Applied Chemistry for Engineering
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    • v.22 no.6
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    • pp.599-604
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    • 2011
  • In this study, we investigated the efficacy of ${\beta}$-cyclodextrin (${\beta}CD$) hydrorogel containing silk fibroin (SF) on healing burnt wound. Tosyl ${\beta}CD$ was conjugated to polyethyleneimine (PEI) using epichlorohydrin (EPI) as a cross-linker. The ${\beta}CD/PEI/SF$ hydrogel was applied on the back of mouse and then the efficacy of hydrogel was compared with both positive control group and negative control group. There was no wound healing efficacy showed neither in the drug loaded ${\beta}CD/PEI/SF$ hydrogel group nor in the drug unloaded ${\beta}CD/PEI/SF$ hydrogel group. On the other hand, in the positive control group, a significant reduction of the wound size after the usage of OTC hydrorogel was obtained. The burn-healing histological result showed a similar phenomenon. After hematoxylin-eosin staining the skin induced by burning, and the epithelial growth observed in the dermis, the efficacy of ${\beta}CD/PEI/SF$ hydrogel in healing burnt wound could not be clearly identified.

Application of Polymerase Chain Reaction (PCR) to the Diagnosis of Tuberculosis (Polymerase Chain Reaction (PCR)을 이용한 결핵의 진단에 관한 연구)

  • Kim, Ho-Joong;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.6
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    • pp.517-525
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    • 1992
  • Background: Since its development by Saiki et al, polymerase chain reaction (PCR) has been very useful in various fields of molecular biology. PCR can be used for the detection of a very small amount of microbial agent, and is especially useful in those patients who are difficult to diagnose microbiologically or serologically. Mycobacterium tuberculosis is a very slowly growing organism and AFB staining frequently shows false negative results, and therefore PCR would be a very rapid, easy, and sensitive diagnostic method for the diagnosis of Mycobacterium tuberculosis. Method: To compare PCR with conventional methods in diagnosing Mycobacterium tuberculosis in sputum, we used sputa of patients who visited or were admitted to Seoul National University Hospital. The amplification targets were 383 base pair DNA, a part of 2520 base pair DNA encoding 65 kD Mycobacterium tuberculosis specific protein (the primers are TB-1, -2), and 123 base pair DNA, a part of IS6110 fragment, which multiple copies are known to exsist PCR one genome (the primers are Sal I-1, -2). We also requested AFB staing and culture to the lab of Seoul National University Hospital with the same sample and compared the results. Results: 1) Using TB-1, -2 primers, PCR was positive in 73.1% (19/26) of culture positive sputa, in 12.5% (1/8) of culture negative. but clinically diagnosed tuberculous sputa, and was negative in all sputa of patients who were clinically diagnosed as non-tuberculous etiology. 2) Using Sal I-I, -2 primers, PCR was positive in 94.1% (32/34) of culture positive sputa, in 23.1% (6/26) of culture negative, but clinically diagnosed tuberculous sputa, and was negative in 87.5% (14/16) of sputa from patients who were clinically diagnosed as non-tuberculous etiology. Conclusion: PCR could be a very rapid, sensitive and specific method for the diagnosis of Mycobacterium tuberculosis in sputa, and further studies should be followed for the development of easier method.

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Diagnostic Significance of the Serologic Test Using Antigen of Mycobacterium Tuberculosis for Antibody Detection by ELISA (결핵항원에 대한 혈청학적 검사와 진단적 유용성)

  • Park, Jae-Min;Park, Yeon-Soo;Chang, Yeon-Soo;Kim, Young-Sam;Ahn, Kang-Hyun;Kim, Se-Kyu;Chang, Joon;Kim, Sung-Kyu;Lee, Won-Young;Cho, Shang-Rae
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.2
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    • pp.271-279
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    • 1998
  • Background: Diagnosis by direct microscopy and/or by culture of the Mycobacterium tuberculosis from body fluids or biopsy specimens is "Gold standard". However, the sensitivity of direct microscopy after Ziehl-Neelsen staining is relatively low and culture of mycobacteria is time consuming. Detection of mycobacterial DNA in clinical samples by the polymerase chain reaction is highly sensitive but laborious and expensive. Therefore, rapid, sensitive and readily applicable new tests need to be developed. So we had evaluated the clinical significance of serologic detection of antibody to 38 kDa antigen, which is known as the most specific to the M. tuberculosis complex, and culture filtrate antigen by ELISA in sputum AFB smear negative patients. Method: In this study, culture tests for acid fast bacilli with sputa or bronchial washing fluids of 183 consecutive patients who were negative of sputum AFB smear were performed. Simultaneously serum antibodies to 38 kDa antigen and unheated culture filtrate of M. tuberculosis were detected by an ELISA method. Results: The optical densities of ELISA test with 38 kDa and culture filtrate antigen were significantly higher in active pulmonary tuberculosis cases than in non tuberculous pulmonary diseases (p<0.05), but in patients with active pulmonary tuberculosis, those of the sputum culture positive patients for M. tuberculosis were not significantly different from those of the sputum culture negative cases(p>0.05). In the smear-negative active pulmonary tuberculosis patients, the sensitivity of the ELISA using 38 kDa antigen and culture filtrate was 20.0% and 31.4%. respectively. The specificity was 95.3% and 93.9%. respectively. Conclusion : In active pulmonary tuberculosis but smear negative, the serologic detection of antibody to 38 kDa antigen and culture filtrate by ELISA cannot substitute traditional diagnostic tests and does not have clinically significant role to differenciate the patient with active pulmonary tuberculosis from other with non-tuberculous pulmonary diseases.

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A STUDY ON CHONDROGENIC POTENTIAL IN MANDIBULAR AND LIMB BUD MESENCHYMAL CELLS OF HUMAN EMBRYOS : A POSSIBLE ROLE OF PROTEIN KINASE C

  • Kook, Yoon-Ah;Kim, Eun-Cheol;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.26 no.6
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    • pp.667-676
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    • 1996
  • We have examined the in vitro stage-related chondrogenic potential of human mandibular and limb bud mesenchyme cells using micromass culture. Our results indicate that limb bud mesenchyme cells as early as stage 16 by Carnegie system (37 days), well before the initiation of in vivo chondrogenesis, have chondrogenic potential which is expressed in micromass culture. These results are correlated with stage-related chondrogenic potential of human limb bud in vivo as a result of Alcian blue staining. The proliferation of chondrogenic cells increased in the first 3 days after culture and then decreased. These results were correlated with the cell cycle analysis of which the number of $G_0^1/G_1$ phase increased markedly after 3 days of culture, while the percentage of cells in S phase was decreased. On the other hand, it was rarely differentiated in the mandible. We examined the effects of two PKC modulators such as phorbol 12-myristate 13-acetate (PMA), a potent activator of PKC, and staurosporine (STSN), an inhibitor of PKC. PMA inhibited the chondrogenesis, whereas STSN promoted the chondrogenesis in a dose dependent manner. In addition, PMA exerted no inhibitory effect when the cells were pretreated for 24 h with STSN, implying that the chondrogenic events might be settled at an early step in vitro and FKC may act as a negative modulator. Collectively, these results demonstrate, for the first time, the stage-related chondrogenic potential of human mandibular and limb bud mesenchyme cells and the role of PKC during chondrogenesis in vitro & in vivo.

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A Case Report of Patient with Left Neck Mass and a History of Thyroid and Ovarian Cancer in Head and Neck (갑상선암 및 난소암 병력을 가진 좌측 경부 종물을 주소로 내원한 환자 1례에 대한 증례 보고)

  • Jeong, Yong Jun;Oh, Kyoung Ho;Kwon, Soon Young
    • Korean Journal of Head & Neck Oncology
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    • v.36 no.2
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    • pp.73-77
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    • 2020
  • Ovarian cancer is common malignant disease with high mortality in the female. However, lymph node metastasis in the head and neck of ovarian cancer is very rare than in para-aortic, pelvic lymph node. A 49-year-old female patient came to our clinic with a left neck mass. After total thyroidectomy and left selective neck dissection for the cervical neck level II, III, IV, V, VI for ovarian cancer and thyroid cancer, she had already undergone chemotherapy (Paclitaxel+Carboplatin) 18 month ago. CT scan showed only lymph node enlargement in left neck level II. Positron emission tomography-computed tomography (PET-CT) revealed a hypermetabolic lesion in same area but no other hypermetabolic lesion, especially in the pelvic and abdominal cavity. Fine needle aspiration cytology revealed metastatic carcinoma. The serum level of CA-125 was elevated to 43.8U/mL, whereas other tumor markers (CA 19-9, CEA) were in the normal range. She underwent a revision of selective neck lymph node dissection for the cervical neck levels I, II, and III, and on the review of surgical pathology, metastatic carcinoma was suspected. Thus, we performed immunohistochemical staining for the tissue; as a result, it was finally diagnosed as metastatic ovarian cancer (positive for CK7, ER and PR, and negative for CK20). Adjuvant chemotherapy (Paclitaxel+Carboplatin) was planned on the tumor board, and the patient successfully received chemotherapy.