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        Effects of epigallocatechin gallate on CoCl_(2)-induced apoptosis in PC12 cells

        Yang, Kyu-Ho,Mo, Hyun-Chul,Choi, Nam-Ki,Kim, Seon-Mi,Kim,Won-Jae 大韓小兒齒科學會 2006 大韓小兒齒科學會誌 Vol.33 No.1

        Neuronal apoptotic events, consequently resulting in neuronal cell death, are occurred in hypoxic/ischemic condition. This cell death has been shown to be accompanied with the production of reactive oxygen species (ROS), which can attack cellular components such as nucleic acids, proteins and phospholipid. However, the underlying mechanisms of apoptosis induced in hypoxic/ischemic condition and its treatment methods are unsettled. Cobalt chloride (CoCl_(2)) has been known to mimic hypoxic condition including the production of ROS. Epigallocatechin gallate (EGCG). a green tea polyphenol, has diverse pharmacologial activities in cell growth and death. This study was aimed to investigate the apoptotic mechanism by CoCl_(2) and effects of EGCG on CoCl_(2)-induced apoptosis in PC12 cells. Administration of CoCl_(2) decreased cell survival in dose- and time-dependent manners and induced genomic DNA fragmentation. Treatment with 100 µM EGCG for 30 min before PC12 cells were exposed to 150 µM CoCl_(2), being resulted in the cell viability and DNA fragmentation being rescued. CoCl_(2) caused morphologic changes such as cell swelling and condensed nuclei, whereas EGCG attenuated morphologic changes by CoCl_(2). EGCG suppressed the apoptotic peak and a loss of Δψ_(m) induced by CoCl_(2). CoCl_(2) decreased Bcl-2 expression but Bax expression was not changed in CoCl_(2)-treated cells. EGCG attenuated the Bcl-2 underexpression by CoCl_(2). CoCl_(2) augumented the cytochrome c release from mitochondria into cytoplasm and increased caspase-8, -9 and caspase-3 activity, a marker of the apoptotic executing stage. EGCG ameliorated the incruement in caspase-8, -9 and -3 activity, and cytochrome c release by CoCl_(2). NAC (N-acetyl-cysteine), a scavenger of ROS, attenuated CoCl_(2)-induced apoptosis in consistent with those of EGCG. These results suggest, that CoCl_(2) induces apoptotic cell death through both mitochondria- and death receptor-dependent pathway and EGCG has neuroprotective effects against CoCl_(2)-induced apoptosis in PC12 cells. 신경세포자멸사는 저산소 및 허혈환경에서 일어나며 이러한 세포죽은 reactive oxidant species (ROS) 생성을 동반함이 알려져있다. 그러나, 저산소 및 허혈환경에서 일어나는 세포자멸사의 기전 및 그 치료방법은 아직 정립되어 있지 않다. CoCl_(2)는 ROS를 생성하는 등 저산소환경과 유사한 조건을 초래하는 것으로 알려져 있다. Epigallocatechin gallate (EGCG)는 녹차의 polyphenol 성분으로서 세포성장과 죽음에 다양한 약리학적 효과를 나타냄이 알려져 있다. 본 연구는 PC12 세포에서 CoCl_(2)에 의한 세포자멸사기전을 밝히고 이에 미치는 EGCG의 효과를 조사하는데 목적이 있다. Cell viability는 MTT 측정으로 조사되었고, DNA fragmentation은 DNA laddering으로 조사되었다. Bcl-2와 Bax발현 정도는 RT-PCR로, caspase-3와 -9의 활성은 spectrophotometer, caspase-8의 활성은 flow cytometry에 의해 측정되었다. 미토콘드리아에서 세포질로 분비된 cytochrome c는 western blot으로, 분해된 DNA 양과 미토콘드리아 세포막전위 (Δψ_(m))는 FACScan으로 조사되었다. CoCl_(2)투여로 PC12 세포수는 용량 및 시간 의존형태로 감소하였고, genomic DNA fragmentation이 발생하였다. CoCl_(2)투여로 야기된 cell viability의 감소와 DNA fragmentation은 EGCG 전처치에 의해 억제되었다. CoCl_(2)은 세포용적팽창과 condensed nuclei 같은 형태적 변화를 일으켰으며, apoptotic peak, Δψ_(m)감소 및 cytochrome c 유리를 야기하였다. EGCG는 CoCl_(2)에 의한 세포형태변화, apoptotic peak, Δψ_(m)소실 및 cytochrome c 유리를 억제시켰다. CoCl_(2)는 Bcl-2 발현을 감소시켰지만, Bax 발현에는 영향을 미치지 않았다. EGCG는 CoCl_(2)에 의해 야기된 Bcl-2 발현 감소를 억제시켰다. CoCl_(2)는 caspase-3, -8, 그리고 -9의 활성을 증가시켰으며, EGCG는 CoCl_(2)에 의한 세포자멸사를 억제시켰다. 본 실험결과는 PC12 세포에서 CoCl_(2)가 미토콘드리아 의존 및 death receptor의존 기전으로 세포자멸사를 일으키며, EGCG는 세포자멸사기전을 억세지킴으로 신경보호기능을 가짐을 시사하였다.

      • 소아 알레르기비염의 경제적 부담 평가를 위한 다기관 조사

        공도연 ( Do Youn Kong ),김경원 ( Kyung Won Kim ),김우경 ( Woo Kyung Kim ),민택기 ( Taek Ki Min ),박용민 ( Yong Mean Park ),안재억 ( Jae Ouk Ahn ),양현종 ( Hyeon Jong Yang ),염혜영 ( Hye Yung Yum ),윤혜선 ( Hae Sun Yoon ),전유훈 ( 대한소아알레르기호흡기학회(구 대한소아알레르기 및 호흡기학회) 2012 소아알레르기 및 호흡기학회지 Vol.22 No.2

        목적: 알레르기비염의 유병률은 전 세계적으로 급격히 증가하고 있으며, 그에 따른 경제적 부담도 증가하고 있다. 그러나, 소아 알레르기 질환이 미치는 경제적 부담에 관한 국내 연구는 제한적이어서 소아 알레르기비염이 환자와 그 가족에게 미치는 경제적 부담을 산출하고자 본 연구를 진행하였다. 방법: 2008년 7월 1일부터 9월 31일까지 서울시 6개 2-3차 의료기관을 방문한 18세 이하 소아 알레르기비염 환자 및 보호자를 대상으로 설문 조사를 진행하였다. 설문을 통해 직접 의료비 (병·의원 진료비, 및 약제비 한방 진료비 및 약제비, 보완/대체요법비), 직접비의료비 (교통비, 환경개선비), 그리고 간접비용 (월 평균 노동 손실)을 조사하였고 그 결과를 질환의 중증도 및 유병 기간에 따라 분석하여 그 차이를 비교하였다. 결과: 모집된 262명의 대상 중 174명(66.4%)이 남자였고, 평균 연령은 6.54세였다. 대상군의 연간 평균 직접 의료비는 177만 원이었고 직접비 의료비는 57만 원이었다. 비록 통계학적 유의성은 없었지만, 알레르기비염의 중증도가 증가할수록 직접 의료비가 증가하는 경향을 보였고, (P=0.053) 유병 기간 또한 직접 의료비의 증가와 유의한 양의 상관 관계를 보였다. (R=0.195, P=0.002) 대상 환자 보호자의 약 17%가 아이의 알레르기비염으로 인해 직장에 결근 또는 조퇴를 경험한 것으로 조사되어 사회적 간접비용을 미루어 생각할 수 있었다. 결론: 알레르기비염이 미치는 경제적 부담은 질환의 중증도가 심할수록, 그리고 유병 기간이 길수록 증가하며, 이는 특히 한방 진료 및 치료비와 보완/대체요법비의 증가에 기인한 것으로 조사되었다. 따라서 알레르기비염으로 인한 경제적 부담을 줄이기 위해서는 환자와 보호자를 대상으로 근거 중심의 치료를 할 수 있도록 지속적이고 체계적인 교육이 필요할 것으로 사료된다. Purpose: The prevalence of allergic rhinitis is rapidly increasing and results in relatively high socio-economic burden on their family and community. However, studies on the economic burden of pediatric allergic rhinitis in Korea are limited. Therefore, we conducted this study to investigate the impact of pediatric allergic rhinitis on economic burden. Methods: Two hundred sixty two children with allergic rhinitis were enrolled in 6 secondary or tertiary medical centers in Seoul from July to September, 2008. We collected data of the economic burden of allergic rhinitis (direct medical costs, direct nonmedical costs, and indirect costs) by face to face questionnaire survey. We compared the economic burden according to the severity and the duration of allergic rhinitis. Results: The mean age of subjects was 6.54 years, and male were 174 (66.4%). Direct medical costs (10,000 Korean Won/yr) were 177.75, and direct nonmedical costs were 57.92. Although, there was no statistical significance, direct medical costs showed increasing trends in severe allergic rhinitis. (P=0.053) In addition, direct medical costs were positively correlated with duration of allergic rhinitis.( R=0.195, P=0.002). About 17% of the parents who care the allergic rhinitis children experienced the work absence due to their child s illness. Conclusion: The economic burdens of allergic rhinitis were positively correlated with the severity and duration of illness. Particularly costs for alternative medicine including oriental medicine` were related with severity and duration allergic rhinitis. Therefore, special efforts for education with evidence based treatment strategy are necessary to decrease the economic burden of allergic rhinitis.

      • Assessment of tissue characteristics of noncalcified coronary plaques by 64-slice computed tomography in comparison with integrated backscatter intravascular ultrasound

        Yang, Woo-In,Hur, Jin,Ko, Young-Guk,Choi, Byung-Wook,Kim, Jung-Sun,Choi, Donghoon,Ha, Jong-Won,Hong, Meonong-Ki,Jang, Yangsoo,Chung, Namsik,Shim, Won-Heum,Cho, Seung-Yun Lippincott Williams Wilkins, Inc. 2010 Coronary artery disease Vol.21 No.3

        BACKGROUND: The ability of 64-slice computed tomography (CT) angiography to differentiate plaque types remains unclear. We evaluated whether the density of noncalcified coronary plaques by 64-slice CT angiography correlates with plaque components assessed by integrated backscatter intravascular ultrasound (IB-IVUS). METHODS: Eighty-six patients [stable angina/acute coronary syndrome (ACS) 67/19, mean age 62±11 years] who showed significant coronary artery stenosis (≥50% diameter stenosis) by 64-slice CT angiography underwent coronary angiography and were evaluated using IB-IVUS. RESULTS: A total of 92 noncalcified coronary plaques on CT angiography were evaluated with IB-IVUS. There was a positive correlation between CT density and calcified tissue content (r=0.41, P<0.001). However, the CT density of plaques did not correlate with other tissue components. Patients with ACS showed more lipid (43.1±13.2 vs. 35.8±13.5, P=0.03) and less soft fibrous tissue (50.5±11.7 vs. 56.5±12.0, P=0.05) by IB-IVUS than those with stable angina. However, the mean CT density of plaques in ACS was not different from that in stable angina (140.6±88.5 vs. 113.1±80.9, P=0.19). CONCLUSION: Except for calcified tissue, CT angiography failed to differentiate plaque types of noncalcified tissue. Therefore, the role of 64-slice CT angiography in identifying lipid-rich plaques remains limited.

      • NADPH oxidase 2 interaction with TLR2 is required for efficient innate immune responses to mycobacteria via cathelicidin expression.

        Yang, Chul-Su,Shin, Dong-Min,Kim, Ki-Hye,Lee, Zee-Won,Lee, Chul-Ho,Park, Sung Goo,Bae, Yun Soo,Jo, Eun-Kyeong Williams Wilkins 2009 JOURNAL OF IMMUNOLOGY Vol.182 No.6

        <P>Gp91(phox)/NADPH oxidase (NOX) 2 is the main catalytic component of NOX, which mediates the phagocytic killing of ingested pathogens via the production of reactive oxygen species (ROS). However, Mycobacterium tuberculosis (Mtb) is relatively resistant to the microbicidal effects of ROS. Thus, the exact roles of NOX2 in the innate immune control against Mtb infection are not fully resolved. In this study, we show that NOX2 is essential for TLR2-dependent inflammatory responses and 1,25-dihydroxyvitamin D(3) (1,25D(3))-mediated antimicrobial activity against Mtb via cathelicidin expression. NOX2-null macrophages prominently abrogated Mtb-induced ROS production and inflammatory signaling activation in a TLR2-dependent manner. Mtb triggered a physical association between NOX2 and TLR2. In addition, the knockdown of NOX2 inhibited 1,25D(3)-triggered antimicrobial activity against viable Mtb through the modulation of cathelicidin expression in human macrophages. Treatment of NOX2 knocked down cells with cathelicidin restored the 1,25D(3)-induced antimicrobial effect, suggesting that the NOX2-dependent induction of cathelicidin in macrophages is part of a defensive strategy against Mtb. Furthermore, cathelicidin expression was required for the Mtb-induced release of ROS and the production of proinflammatory cytokines/chemokines, indicating a positive circuit of inflammation in response to Mtb. Our data collectively demonstrate a novel regulatory mechanism for TLR2-dependent innate responses to Mtb involving crosstalk between NOX2 and TLR2 and the expression of cathelicidin.</P>

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        Energy And Environmental Engineering : Headspace Solid-Phase Microextraction for Determination of Micellar Solubilization of methyl tert-butyl ether (MTBE)

        ( Ki Tae Baek ),( Hyun Ho Lee ),( Hyun Jeong Cho ),( Ji Won Yang ) 한국화학공학회 2003 Korean Journal of Chemical Engineering Vol.20 No.4

        Headspace analysis using solid-phase microextraction (SPME) was tested as a rapid method to evaluate micellar solubilization of methyl tert-butyl ether (MTBE) with sodium dodecyl sulfate (SDS) and cetylpyridinium chloride (CPC). At equilibrium between aqueous phase and vapor phase, free MTBE in vapor phase (i.e., not solubilized MTBE by SDS or CPC) was analyzed by GC-FID (Hewlett Packard 5890 series II) equipped with a capillary HP1 column (30 m×0.25 mm). This method showed a good analytical performance such as linearity of calibration curve and precision (RSD less than 5%). Extent of MTBE solubilization was expressed as a function of molar ratio of SDS to MTBE and CPC to MTBE, and was saturated at about 57% and 37% with over the value of ratio 13 for SDS and CPC, respectively. This technique can be applied to analyze micellar solubilization potential of various surfactants on volatile and semi-volatile compounds.

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