• 제목/요약/키워드: Immunofluorescent

검색결과 178건 처리시간 0.029초

Vasodilation of BCT is Associated with Inhibition of PKC$\alpha$ Translocation and LC20 Phosphorylation

  • Kwon Oh Kui;Shin Dong Hoon;Kim Gil Whon;Shin Heung Mook
    • 동의생리병리학회지
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    • 제17권5호
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    • pp.1335-1338
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    • 2003
  • We have previously reported that the vasodilatory effect of BanhabackchuIChunma-tang(半夏白朮天麻湯; BCT), a herbal formula, and its mechanism might be associated with at least in part NO pathway. In the present study, we studied the influence of BanhabackchulChenuma-tang (BCT) on the phosphorylation of LC20, in parallel, the distribution of α-protein kinase C(PKCα) by phenylephrine was monitored using laser scan confocal immunofluorescent microscopy in freshly isolated ferret portal vein smooth muscle living single cells. Phenylephrine stimulation induced LC20 phosphorylation and translocation of PKCα. However, BCT dephosphorylated LC20 phosphorylation and inhibited the translocation of PKCα. Our results demonstrate that the mechanism of relaxant effect of BanhabackchulChunma-tang inhibition is associated with inhibition of PKCα activation and LC20 phosphorylation.

소폐동맥 내피세포를 이용한 인조혈액접촉표면의 혈액 접합성 (Blood Compatibility of Artificial Blood-Contacting Surface Seeded with Cultured Bovine Endothelial Cells)

  • 김원곤;곽영태;유세영
    • Journal of Chest Surgery
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    • 제26권2호
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    • pp.80-85
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    • 1993
  • Synthetic and biosynthetic vascular grafts of small diameter have long been considered to be prone to thrombosis, ultimately leading to the complete graft occlusion. Endothelial cell seeding onto synthetic blood-contacting surfaces has been suggested to be an ideal means to solve this problem. This study described a culture method of bovine endothelial cells and evaluated blood-compatibility and seeding efficiency of cultured endothelial cells. Bovine pulmonary artery endothelial cells were harvested enzymatically and grown to confluence on polystyrene culture flask surfaces using established techniques. The identification of endothelial cells was made through the demonstration of expression of factor VIII R:Ag by immunofluorescent technique. To quantitate the effect of improvement in blood-compatibility of viable endothelial cells, endothelial monolayers were exposed to blood containing $\^$111/In-oxine labeled platelets. Viable endothelial monolayers retained less labeled platelets than control surfaces. The Indium-labeled endothelial cells were seeded onto three different blood-contacting surfaces of Dacron vascular graft immobilized in specially equipped wells and incubated for specific time intervals (t=15, 30, 60, 120 minutes). Longer incubation times showed improved cell adherence in collagen-coated and fibrin-coated Dacron vascular graft groups. However in untreated Dacron grafts, no direct relationship was observed between incubation time and endothelial cell seeding efficiency. This may be due to leakage of endothelial cells through porosity of Dacron grafts in this in-vitro experimental condition.

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한국산 다람쥐의 Orientia Tsutsugamushi에 대한 항체양성율 (Seropositive Rate of Orientia Tsutsugamushi in Tamias Sibiricus from Korea)

  • 송진원;이상원;고은영;정기모;이용주;송기준;백락주
    • 대한미생물학회지
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    • 제35권1호
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    • pp.61-68
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    • 2000
  • Among wild chipmunks, Tamias sibiricus, captured in Kyunggi and Kangwon province in Korea, 1997, seropositivity for Orientia tsutsugamushi was determined. Serological test for Orientia tsutsugamushi infection was performed using indirect immunofluorescent antibody technique (IFA). Of 243 wild chipmunks, 61 against Gilliam strain and 64 against Karp strain of Orientia tsutsugamushi were IFA positive. Seropositivity against Gilliam strain was shown 33.3% in Kyunggi and 23.5% in Kangwon province, and against Karp strain was shown 33.3% and 25.4%, respectively.

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구강 편평상피세포암 동위종양 모델에서 내피세포의 수용체 타이로신 인산화효소에 대한 표적치료 (TARGETING RECEPTOR TYROSINE KINASE ON ENDOTHELIAL CELLS IN AN ORTHOTOPIC TUMOR MODEL OF ORAL SQUAMOUS CELL CARCINORMA)

  • 박영욱;김소희
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제35권2호
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    • pp.55-65
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    • 2009
  • Purpose: We determined the therapeutic effects of blockade of epidermal growth factor(EGF) and vascular endothelial growth factor(VEGF) receptor tyrosine kinases on the growth of oral squamous cell carcinoma(OSCC) xenografted in athymic nude mice. Experimental Design: We investigated the in vivo antitumor effects of a tyrosine kinase inhibitor for EGFR and VEGFR-2, AEE788 in a mouth floor(orthotopic) tumor model. Nude mice with orthotopic tumors were randomized to receive AEE788, paclitaxel, a combination of AEE788 and paclitaxel, or control. Antitumor mechanisms of AEE788 were determined by immunohistochemical/immunofluorescent and apoptosis assays. Results: Tumors of mice treated with AEE788 demonstrated down-regulation of phosphorylated EGFR, phosphorylated VEGFR and their downstream mediators(pMAPK and pAkt), decreased proliferative index, decreased microvessel density(MVD). As a result, growth of the primary tumor and nodal metastatic potentials were inhibited by AEE788. Conclusion: These data show that EGFR and VEGFR can be molecular targets for the treatment of OSCC.

혈관내피성장인자의 섬유아세포 증식과 Notch 1 발현에 대한 영향 (Vascular Endothelial Growth Factor Effect on Notch 1 Expression and Proliferation of Fibroblast)

  • 고성훈
    • Archives of Plastic Surgery
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    • 제37권1호
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    • pp.7-11
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    • 2010
  • Purpose: Vascular endothelial growth factor (VEGF) is known as a growth factor of endothelium and fibroblast. The purpose is to know the VEGF effects on fibroblast proliferation and fibroblast's notch receptor expression. Methods: CCD-986sk fibroblast was purchased from the Korean Cell Bank and was used in XTT assay for proliferation and wound healing assay for migration. Immunofluorescent (IF) staining and western blotting were used in testing notch expression of fibroblast. Semiquantitative RT-PCR was used in checking notch 1 mRNA production by fibroblast. Student-t test was used for analyzing results. Results: Cell proliferation assay using XTT showed significant higher proliferation in VEGF treated fibroblast, $2.324{\pm}0.0026$ vs. $2.463{\pm}0.017$ (p=0.002). Wound healing assay showed longer migration in VEGF treated fibroblast (p=0.062). The fluorescence was brighter in VEGF treated cells of notch 1 IF staining. Notch 1 expressions and mRNA productions increased more in VEGF treated cells. Conclusion: VEGF stimulates fibroblast to proliferate, migrate and to express Notch 1 simultaneously. Notch receptor could be related to VEGF mediated wound healing.

돼지 생식기 호흡기 증후군 바이러스 자연감염 예의 병리학적 연구 (Pathologic Studies in Piglets Naturally Infected with Porcine Reproductive and Respiratory Syndrome Virus)

  • 김재훈;황의경;김용주;손현주
    • 한국수의병리학회지
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    • 제1권2호
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    • pp.125-134
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    • 1997
  • Porcine Reproductive and Respiratory Syndrome Virus infection (PRRSV) was confirmed by serology histopathology immunohistochemistry and bacteriologic examination in young pigs. Four suckling and six weaned piglets submitted from three different farms showed coughing sneezing labored rapid abdominal respiration lethargy and anorexia. Grossly apical and cardiac lung lobes appeared mottled with pale to dark tan discoloration. Submandibular and bronchial lymph nodes were tan and enlarged. All piglets were seropositive for PRRSV antibodies by the indirect immunofluorescent antibody(IFA) test. Microscopically lung lesions were characterized by hyperplasia and hypertrophy of type 2 pneumocytes infiltration of mononuclear cells in alveolar intersitium accumulation of necrotic debris in alveolar spaces accompanied by proliferation of alveolar multinucleated syncytial cells. Using immunohistochemical technique PRRSV antigens were demonstrated in alveolar macrophages and type 2 pneumocytes in histologic lung tissue sections. Also PRRSV antigens were detected in brain lymph nodes spleen and heart. Additionally piglets showed nonsuppurative meningoencephalitis mandibular necrotic lymphadenopathy splenic atrophy and myocardial necrosis.

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갑상선 종양에 있어서 말초혈의 T 세포 Subset에 관한 연구 (A Study on Peripheral T Cell Subsets in Patients with Thyroid Tumors)

  • 김동수
    • 대한핵의학회지
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    • 제25권1호
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    • pp.110-116
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    • 1991
  • To elucidate alteration of peripheral T cell subsets in thyroid tumors, the author enumerated T cell subsets in periphral blood by indirect immunofluorescent method, using monoclonal antibodies (CD3, CD4 and CD8) in 17 cases of thyroid cancer, 12 cases of thyroid adenoma, and 16 cases of adult healthy subjects as controls. Diagnoses were confirmed histopatologically in thyroid cancer and adenoma, and were established on the basis of commonly accepted clinical and biochemical criteria in Hashimoto's thyroiditis. The blood was drawn from veins of the patients and control subjects in Pusan National University Hospital during the period of January to October 1990. The results obtained were summarized as follow: 1) The percentage of CD3+ cells was significantly decreased in thyroid cancer as compared with healthy subjects. 2) The percentage of CD4+ cells was not different among thyroid cancer, thyroid adenoma, Hashimoto's thyroiditis and control subjects each other. 3) The percentage of CD8+ cells was significantly decreased in thyroid cancer as compared with adult healthy subjects, and tended to be decreased as compared with thyroid adenoma and Ha-shimoto's thyroiditis. 4) The CD/CD8 ratio was significantly increased in thyroid cancer as compared with control subjects, and tended to be increased as compared with thyroid adenoma and Hashimoto's thyroiditis. On the basis of the results, it can be suggested that the immunodysfunction may be due to decreased soppressor/cytotoxic T cells in thyroid cancer.

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한타바이러스 혈청형 특이 Primer를 이용한 Nested RT-PCR 방법으로 5가지 혈청형 한타바이러스에 감염된 햄스터 조직에서 바이러스 검출 (Discrimination of Hantaviruses from the Tissues of Infected Hamsters to 5 Different Serotype Hantaviruses by Nested RT-PCR using Hantavirus Serotype Specific Primers)

  • 주용규;이호왕
    • 대한바이러스학회지
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    • 제27권1호
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    • pp.49-57
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    • 1997
  • We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.

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대전지역 길고양이의 톡소포자충(Toxoplasma gondii) 감염 실태 조사 (Investigation of Toxoplasma gondii infection on stray cats in Daejeon)

  • 성선혜;유상식;임여정;정년기;문병천
    • 한국동물위생학회지
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    • 제35권1호
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    • pp.19-24
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    • 2012
  • This study was performed to evaluate the prevalence rate of Toxoplasma gondii on 217 stray cats in Daejeon. The positive infection rate of T. gondii was 15.7% in enzyme-linked immunosorbent assay (ELISA), 12.4% in latex agglutination test (LAT), 14.7% in indirect immunofluorescent antibody test (IFA) and 0.5% in polymerase chain reaction (PCR) respectively. In districts, Yuseong-gu was shown the highest seropositive rate of T. gondii as 31.8% in ELISA, 22.7% in LAT and 31.8% in IFA. In gender, the seropositive rate of female cats was slightly higher than that of male cats as 17.2% in ELISA, 15.2% in LAT, 15.2% in IFA and 1.0% in PCR. Cats captured in National science museum, detached house and apartment was shown relatively high prevalence rate of T. gondii.

국내 야생 집쥐(Rattus norvigicus)의 유행성 출혈열바이러스 감염증의 역학조사 (Epidemiological survey on Hantaan-related virus infection of wild rats(Rattus norvigicus) in Korea)

  • 김희선;;이준학;강문일;고홍범
    • 대한수의학회지
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    • 제33권2호
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    • pp.321-326
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    • 1993
  • Sixty eight wild rats were caught from Seoul, Kyonggi, Kangwon, Cholla, and Kyongsang provinces in Korea. All of them were confirmed in the same species, Rattus norvigicus. All of sera from wild rats were examined by indirect immunofluorescent test against Hantaan-related virus. Detection of anti-body to Hantaan-related virus were 4 out of 68 rats(5.9%), including 2 of 8(25.0%) rats in Kangwon and 2 of 20 rats(10.0%) in Kyongsang province. No difference of the sero-positive rates between sex of rats tested to Hantaan-related virus was recognized. According to age, four rats with antibodies(1:320~640) to Hantaan-related virus belonged one subadult, one young adult, one middle aged adult, and one old adult, respectively.

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