• Title/Summary/Keyword: 면역세포화학 염색법

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Expression of Epidermal Growth Factor Receptor in the Inflamed Gingival Epithelium and the Dental Follicle (염증성 치은 상피와 치낭의 표피성장인자 수용체의 발현 및 실험적 치아이동에 미치는 영향에 관한 연구)

  • Kim, Young Ho;Bae, Chang
    • The korean journal of orthodontics
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    • v.27 no.2
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    • pp.349-357
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    • 1997
  • Epidermal growth factor(EGF), a single chain polypeptide of 53 amino acids with a molecular weight of 6,045 Da, was first isolated from the male mouse submandibular glands. EGF stimulates cellular proliferation and differentiation in several tissues and accelerates the rate of wound healing. EGF is bound to the specific receptor(EGFR) on the cell membrane of its target cell. EGFR is a transmembrane glycoprotein with a molecular weight of 170,000 Da and is detectable on a large variety of cell types and tissues. The authors investigated the expression of EGFR in the normal and inflamed human gingival epithelium to study the role of EGFR in the inflammation of the gingival epithelium, and the expression of EGFR in the dental follicle by using in situ mRNA hybridization and immunohistochenistry. The results weree as follows : 1. The expression of EGFR mRNA in the normal gingival epithelium on in situ mRNA hybridization was mainly localized on the basal cell layer, and the spinous layer was weakly positive The granular and cornified layers were negative 2. The expression of EGFR protein in the normal gingival epithelium on inmunohistochemistry was localized on the cornified and granular layers, and the spinous layer was weakly positive. The basal cell layer was completely negative 3. The expression of EGFR mRNA in the inflamed gingival epithelium on in situ mRNA hybridization was evenly and homogeneously distributed in the whole layers of the gingival epithelium except the cornified layer. The staining intensity appeared to increase progressively from the basal cell layer to the cornified layer. 4. The expression of EGFR protein in the inflamed gingival epithelium on immunohistochemistry was evenly and homogeneously distributed in the whole layers of the gingival epithelium. The staining intensity appeared to increase progressively from the cornified layer to the basal cell layer. 5. Strong positive reaction was seen in the epithelial cell rests of Malassez, whereas only background staining was seen in other cells of the dental follicle. In conclusion, the up-regulation of EGFR in the inflamed gingival epithelium and the high amounts of EGFR in the epthelial cell rests of Malassez in the dental follicle can be regarded as responses to the possible damages to the oral environment to maintain the homeostatic conditions.

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Characterization of Umbilical Cord-derived Stem Cells during Expansion in Vitro (탯줄유래 줄기세포의 계대배양에 따른 특성 변화의 분석)

  • Park, Se-Ah;Kang, Hyun-Mi;Heo, Jin-Yeong;Yoon, Jin-Ah;Kim, Hae-Kwon
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.1
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    • pp.23-34
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    • 2009
  • Objectives: Mesenchymal stem cells (MSC) comprise a promising tool for cellular therapy. It is known that long-term in vitro culture of human bone marrow and adipose tissue derived-MSCs lead to a reduction of life span and a change of stem-like characters. The aim of our study was to examine whether stem cell properties of human umbilical cord-derived stem cells (HUC) could be affected by in vitro expansion. Methods: HUC were isolated from human umbilical cord and cultured for 10 passages in vitro. Morphology and population doubling time (PDT) were investigated, and changes of stem cell properties were examined using RT-PCR and immunocytochemistry during serial subcultures. Results: Morphology and PDT of HUC began to change slightly from the 7th passage (p7). Expression level of nestin and vimentin mRNAs increased along with the culture period from p4 until p10. In contrast, expression level of SCF mRNA decreased during the same culture period. Expression level of Oct-4 and HNF-4${\alpha}$ mRNAs was not significantly changed throughout the culture period until p10. Expression level of BMP-4, FGF-5, NCAM and HLA-ABC mRNAs appeared to increase as the culture continued, however, the difference was not significant. Immunocytochemical studies showed that HUC at p3, p6 and p9 positively were stained with antibodies against SSEA-3 and SSEA-4 proteins. Interestingly, staining intensity of HUC for ICAM-1 and HLA-ABC gradually increased throughout the culture period. Intensity against thy-1 and fibronectin antibodies increased at p9 while that against TRA-1-60 and VCAM-1 antibodies began to decrease at p6 until p9. Conclusions: These results suggest that HUC change some of their stem cell characteristics during in vitro culture. Development of culture system might be needed for the maintenance of characteristics.

Expression of Estrogen and Progesterone Receptors in Non-small-cell Lung Cancer Tissue Using Tissue Microarray Method (조직 미세배열법을 이용한 비소세포 폐암 조직에서 에스트로겐과 프로게스테론 수용체 발현)

  • Han, Hye-Seung;Kim, Min-Ji;Cho, Jae-Hwa;Yoon, Yong-Han;Kwak, Seung-Min;Lee, Hong-Lyeol;Kim, Kwang-Ho;Ryu, Jeong-Seon
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.1
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    • pp.54-58
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    • 2005
  • Background : To evaluate the role of estrogen and progesterone in the carcinogenesis of NSCLC, IHC studies for the expression of the receptors of estrogen and progesterone have been performed with inconsistent results. Recently the TMA method has been developed and has become recognized as a useful and rapid method for extensively analysing molecular markers at the gene and protein level. We have investigated their expressions in the tissue from NSCLC using the microarray method. Methods : The TMA construction was made with 70 formalinfixed, paraffin-embedded tissues of NSCLC. After heat-induced epitope retrieval, IHC staining on primary tissues of NSCLC was performed with the monoclonal antibodies, ER1D5 and PR1A6. Results : Our sample of 70 consisted of 74% men and 26% women. Of the patients, 49% were current smokers, 27% were non-smokers and 24% were former smokers. By histologic classification, 34 patients had squamous cell carcinoma, 24 had adenocarcinoma, 9 had adenosquamous cell carcinoma, and 3 had other carcinomas. No cancer cells were immunostained with these monoclonal antibodies in any primary tissues of NSCLC. Conclusions : No expression of neither of the two receptors was found in any of the lung cancer tissues. This suggests that adequate genetic variants for IHC staining need to be developed for NSCLC.

Immunohistochemical Detection of p53, erbB-2 and CEA Oncoprotein in Lung Cancer; Clinical Correlations (폐암 환자에서 면역조직화학 염색을 통한 p53, erbB-2, CEA 종양단백 발현과 임상적 의의)

  • Jeong, Seong-Su;Kang, Dong-Won;Lee, Gyu-Seung;Ko, Dong-Seok;Suh, Jae-Chul;Kim, Geun-Hwa;Shin, Kyoung-Sang;Kim, Ju-Ock;Song, Gyu-Sang;Kim, Sun-Young
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.766-775
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    • 1998
  • Background : The prognosis of patients with lung cancer is still poor. Lung cancer exhibits a variable clinical outcome, even in those patients with same stage. Numerous reports suggest that oncogene expression might playa role in explaining the variability of response and survival But many of these reports are still under debate. So we studied the clinical relevance of oncogene expression in Korean lung cancer patients. Immunohistochemistry of p53, erbB-2, CEA expression was performed. Method: From March, 1992 until March, 1997, 120 patients with lung cancer were reviewed. p53, erbB-2, and CEA expression were detected on paraffin-embedded tumor blocks with the use of monoclonal antibodies. The survival and response has correlated with the expressibility of p53, erbB-2, and CEA oncoprotein Results: Overall, the expression rates of p53, erbB-2, and CEA were 33.7%, 59.3%, and 32.6% respectively. Expression rates were not correlated to cell type or stage. Compared with response to chemotherapy, no correlation was found. The expression of p53, erbB-2, or CEA was not correlated with 2-year survival. With simultaneous applications of p53, erbB-2, and CEA, patients with 2 or more expressions also did not show poor response to chemotherapy. Conclusion: We conclude the p53, erbB-2, and CEA expression are clinically less useful in predicting response to chemotherapy or survival.

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Immunohistochemical Staining of Insulin-like Growth Factor-1 in Human Lung Cancer Cells (폐암의 조직학적 형태에 따른 인슐린양 성장인자-1의 면역조직학적 염색의 비교)

  • Park, Ji-Hyun;Kang, Myoung-Jae;Lee, Heung-Bum;Lee, Yong-Chul;Rhee, Yang-Kuen
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.3
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    • pp.324-330
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    • 2000
  • Objective : Lung cancer arises after a series of morphological changes, which take several years to progress from normal epithelium to invasive cancer. Multiple molecular changes and growth factor production have been documented in lung cancers, both small cell and non-small cell types. Insulin-like growth factors(IGFs) are important mitogenic and anabolic peptides, both in vivo and in vitro, and are thought to be significant autocrine-paracrine factors involved in normal and malignant cell proliferation. In this study, the degree of expression of IGF-1 on the immunohistochemical staining in human non-small cell lung cancer(NSCLC) cells and small cell lung cancer (SCLC) cells were investigated. Methods : Immunohistochemical staining for IGF-1 was performed in 15 cases of small cell carcinoma, 15 cases of squamous cell carcinoma, 15 cases of adenocarcinoma, and 12 cases of bronchoalveolar carcinoma. Results : The expression of IGF-1 on the immunohistochemical staining significantly increased in NSCLC cells than in SCLC cells. Conclusion : These results suggest the expression of IGF-1 in human lung cancer cells. The immunohistochemical staining of IGF-1 in lung cancer cell lines may assist in the differentiation of NSCLC and SCLC.

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The Morphologic Changes of Parvalbumin- Immunoreactive Interneurons of the Dentate Gyrus in Kainate-Treated Mouse Hippocampal Slice Culture Epilepsy Model (Kainic Acid로 처리한 해마박편배양 마우스 간질모델에서 치아이랑 Parvalbumin 면역 반응성 사이신경세포의 형태학적 변화)

  • Chung, Hee Sun;Shin, Mi-Young;Kim, Young-Hoon;Lee, In-Goo;Whang, Kyung-Tai;Kim, Myung-Suk
    • Clinical and Experimental Pediatrics
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    • v.45 no.12
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    • pp.1551-1558
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    • 2002
  • Purpose : Loss of hippocampal interneurons in dentate gyrus has been reported in patients with severe temporal lobe epilepsy and in animals treated with kainic acid(KA). Interneurons contain $Ca^{2+}$- binding protein parvalbumin(PV). The effects of kainic acid on parvalbumin-immunoreactive (PV-IR) interneurons in dentate gyrus were investigated in organotypic hippocampal slice cultures. Methods : Cultured hippocampal slices from postnatal day nine C57/BL6 mice were exposed to $10{\mu}M$ KA, and were observed at 0, 8, 24, 48, 72 hours after a one hour KA exposure. Neuronal injury was determined by morphologic changes of PV-IR interneuron in dentate gyrus. Results : Transient(1 hour) exposure of hippocampal explant cultures to KA produced marked varicosities in dendrites of PV-IR interneuron in dentate gyrus and the shaft of interbeaded dendrite is often much thinner than those in control. The presence of varicosities in dendrites was reversible with KA washout. The dendrites of KA treated explants were no longer beaded at 8, 24, 48 and 72 hours after KA exposure. The number of cells in PV-IR interneurons in dentate gyrus was decreased at 0, 8 hours after exposure. But there was no significant difference in 24, 48 and 72 hours recovery group compared with control group. Conclusion : The results suggested that loss of PV-IR interneurons in dentate gyrus is transient, and is not accompanied by PV-IR interneuronal cell death.

The Significance of p53 Expression in Serum and Tissue from Patients with Lung Cancer (원발성 폐암환자의 혈청 및 조직에서의 p53단백 표현)

  • Chang, Jung-Hyun;Sung, Sun-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.2
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    • pp.333-340
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    • 1998
  • Background: Lung cancer is the leading cause of cancer over the world. P53 alteration is by far the most common genetic defect in lung cancer. The mutation of p53 protein involves the loss of inhibitory function of p53 related tumor suppressor gene and resultant oncogenesis. The analysis of p53 alterations consists of immunohistochemical stain, PCR based assay, or serologic ELISA (enzyme-linked immunosorbent assay). Methods : Serum levels of p53 mutant protein were measured in 69 cases of lung cancer (adenocarcinoma n=29, epidermoid n=16, small cell n=13, large cell n=1, undifferentiated n=1, undetermined n=9) and 42 controls of respiratory disorders using ELISA. Immunohistochemical stain in tissue was performed using monoclonal antibody of p53 in lung cancer subjects. Results: Both serum p53s in nonsmall cell cancer ($0.28{\pm}0.44ng/ml$) and in small cell cancer ($0.20{\pm}0.14ng/ml$) were not different from controls ($0.34{\pm}0.20ng/ml$). Also there was no significant difference in serum p53 according to tumor stages. P53 immunohistochemical stain showed 50% positivity overall in lung cancer. There were no close correlation between serologic level and positivity of immunohistochemical stain. Conclusion: The serologic determination of p53 mutant protein is thought to have no diagnostic role in lung cancer. Immunohistochemical stain in lung cancer specimen shows 50% positivity.

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Immunohistochemical Localization of 36 and 29 kDa proteins in sparganum (면역조직염색법으로 관찰한 스파르가눔 층체에서의 36, 29 kDa 항원 단백질의 생성위치)

  • Kim, Lee-Su;Kong, Yoon;Kang, Shin-Yong;Cho, Seung-Yull
    • Parasites, Hosts and Diseases
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    • v.30 no.1
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    • pp.25-32
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    • 1992
  • Antigenic proteins of 36 and 29 kDa were localized in Spirometra mansoni plerocercoid (sparganum) immunohistochemically by avidin biotin complex (ABC) staining. When polyclonal antibodies such as BALB/c mouse serum immunized with crude saline extract of sparganum or confirmed sparganosis sera were reacted as pri-mary antibodies, the positive chromogen (3-amino, 9-ethylcarbazole) reactions were recognized at syncytial tegument, tegumental cells, muscle and parenchymal cells and lining cells of excretory canals. A monoclonal antibody(MAb) which was reacting to 36 and 29 kDa proteins in the extract of the worm was localized at the syncytial tegument and tegumental cells. The present results suggested that the potent antigenic proteins of 36 and 29 kDa in sparganum were produced at the tegumental cells and transported to the syncytial tegument.

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Measurement of Mucin Amounts Using RTO3 in Patients with Chronic Airway Disease (만성 기도질환에서 흰쥐 뮤신에 대한 단 세포군 항체(RTO3)를 이용한 뮤신 측정에 관한 연구)

  • Kim, Do-Jin;Kim, Ki-Up;NamGung, Eun-Kyang;Uh, Soo-Taek;Kim, Young-Hoon;Shin, Chan-Young;Ko, Kwang-Ho;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.6
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    • pp.786-796
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    • 1999
  • Background: It has been anticipated that the amount and composition of mucin are changed in patients with chronic airway diseases. We evaluated whether RTO3(mAb against rat tracheal mucins) could quantify the amount of mucin from the airway in the patients with chronic airway diseases. Methods and results; 1) RTO3 was bound to high molecular weight of mucin based on Western blot in sputum and BALF from patients with chronic airway diseases. 2) The goblet cells and submucosal glands in main bronchus from human were observed by PAS stain. And immunohistochemical stain with RTO3 showed immunoreactivity on some goblet cells. 3) The amount of mucin was more increased in patients with chronic airway diseases compared to those in normal subjects. 4) In the exacerbation of asthmatics, mucin amounts were more increased than stable asthmatics. Conclusion: We suggested that secreted mucin in chronic airway diseases can be quantified by ELISA with RTO3.

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Differential Diagnosis of Pleural Mesothelioma and Metastatic Adenocarcinoma by Immunohistochemistry (면역조직화학염색법을 이용한 흉막의 악성중피종과 전이성 선암의 감별진단)

  • Ko, Kyung-Haeng;Park, Chang-Min;Rim, Myung-Soo;Kim, Yoo-Il;Jang, Il-Gweon;Hwang, Joon-Hwa;Lim, Sung-Chul;Kim, Young-Chul;Park, Kyung-Ok;Park, Chang-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.4
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    • pp.478-487
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    • 1999
  • Background : Differential diagnosis of pleural malignant mesothelioma from secondary metastatic adenocarcinoma is often difficult. A variety of pathologic techniques have been developed to make a differential diagnosis of carcinoma from mesothelioma. Immunohistochemistry detecting diverse antigenic substances such as CEA, Leu-M1, Bn-3, S-100 protein, vimentin, CK and EMA has been claimed to be of value as a panel in the differential diagnosis of adenocarcinoma from mesothelioma. The aim of this study was to investigate the suitable antibodies to distinguish mesothelioma from metastatic adenocarcinoma and establish candidate markers in a panel. Methods : Complete, one-hour immunohistochemical staining using antibodies against cytokeratin (CK), epithelial membrane antigen(EMA), S-100 protein, vimentin, B72-3, Leu-M1, and carcino-embryonic antigen(CEA) was applied to cell blocks from 7 mesotheliomas and 7 adenocarcinomas which were confirmed by electron microscopic and histpathologic methods. Results : All adenocarcinomas and 71.4% of mesotheliomas expressed the cytokeratin and EMA. S-100 protein and vimentin were expressed in 57.1% and 42.9% of mesotheliomas and 14.3% and 28.5% of adenocarcinomas, respectively. B72-3 was expressed in all adenocarcinomas, but in none of mesotheliomas. Leu-M1 was positive in 71.4% of the adenocarcinoma and 14.3% of the mesotheliomas. CEA was positive in all adenocarcinomas and 42.9% of mesotheliomas. Leu-M1 and B72-3 were coexpressed in 71.4% of adenocarcinomas but in none of mesothelioma. B72-3 and CEA were coexpressed in all adenocarcinomas, but in none of mesotheliomas. Conclusion : We concluded that B72-3 immunohistochemistry or panel staining of B72-3 and CEA could be recommanded for the differential diagnosis of pleural mesothelioma from metastatic adenocarcinoma.

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