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SV 40 Large T 유전자를 이용해 불멸화시킨 사람 태아 간세포주의 확립
박중원(Joong Won Park),이주영(Joo Young Lee),이효석(Hyo Suk Lee),박주배(Joo Bae Park),김정룡(Chung Yong Kim) 대한내과학회 1994 대한내과학회지 Vol.46 No.4
Background: Hepatocyte culture represents a cell model for analyzing the mechanism involved in car- cinogenesis of carcinogens, a tool for measuring hepatotoxicity of drugs and a simple model for studying hepatitis virus life cycle. However, problems raised by both the short-term survival and the poor functional stability of hepatocytes in culture hindered scientist in using of this in vitro system. Thus we planed to obtain the immortalized and differentiated human hepatocyte cell line by modifying the genome by transfecting the cells with viral specific DNA, always available in studying the liver disease. Method: After primary culture of hepatocytes obtained by therapeutic abortion at 18 weeks of gestation, SV40 large T gene was transfected into the cells by using Polybren-DMSO method, And then transfected hepatocytes were selected in G418 containing medium. Selected, transformed hepatocytes were subcultured in 10% fetal bovine serum containg F-12 medium. Morphological characteristics of subcultured cells was followed by phase-contrast microscope and electoron microscope. The immunocytochemisty using anti-human albumin and anti-human alpha fetoprotein and the immunofluorescence using snti-SV40 T antigen were performed for proving the differentiation of sub-cultured hepatocytes. Results: Electron microscope revealed subcultured cell to be epithelial cell. After more than 20 passages over a period of 7 months, the cells retained an epitheloid morphology. All the SV40 transformed hepatocyte cell lines were 100% positive for T antigen. Significant anti-alpha fetoprotein staining and week anti-albumin staining were observed in cytoplasm around nucleus and so we confirmed the synthesis of liver specific protein of transformed hepatocytes, Conclusion: Human fetal immortalized hepatocytes cell line secreting the liver specific proteins was established.
Chlorpromazine이 백서 해마에서 전기경련 충격에 의한 MAPK 타이로신 인산화에 미치는 영향
주연호,정성훈,전송희,강웅구,김용식,김상욱,박주배 大韓神經精神醫學會 1997 신경정신의학 Vol.36 No.2
Object : In order to examine the interaction mechanisms of electroconvulsive shock(ECS) and antipsychotic drug at the level of molecular biology, we observed the effect of chlorpromazine pre-treatment on the activation of mitogen activated protein kinase(MAPK) induced by electroconvulsive shock(ECS) in rat hippocampus Method : Male Sprague-Dawley rats were divided into 2 groups. To the experimental group chlorpromazine(20㎎/㎏) was given intraperitoneally, and to the control distilled water was given instead. Thirty minutes later, ECS was given and the hippocampus was dissected out 2 minutes thereafter. Immunoblotting with antiphosphotyrosine antibody was carried out, and the signal intensity at 42kDa band was quantitized using densitometer. The obtained result was compared by student t-test between the experimental and the control group. The absolute amount of MAPK was measured by immunoblotting with anti-MAPK antibody. Result : The tyrosine phosphorylation of MAPK reached peak at 2 minutes after ECS. However, in the chlorpromazine pre-treated group, the peak level of MAPK tyrosine phosphorylation was significantly attenuated(t=-3.12, df=14, p=0.008) compared to the control. In contrast to this, the absolute amount of MAPK did not differ between the pretreated and the control group. Conclusion : Chlorpromazine attenuated the tyrosine phosphorylation of MAPK by ECS. This finding seems to be related to the fact that chlorpromazine pre-treatment changed the cfos expression by ECS in rat brain. Antipsychotic drug and ECS might interact at the level of MAPK signal transduction system, and this might explained the observed synergistic effect of two treatment modality.
정신분열병 환자의 혈청과 뇌척수액의 동 및 아연농도에 관한 예비적 연구
김용식,장안기,박주배 大韓神經精神醫學會 1983 신경정신의학 Vol.22 No.4
27명의 정신분열환자 대상군과 10명의 대조군을 두어 atomic absorption spectrotometry를 사용하여 혈청과 뇌척수액의 동과 아연농도를 측정하여 이를 비교하고, 항정신약물의 미치는 영향을 연구하여 다음과 같을 결론을 얻었다. 첫째, 혈청과 뇌척수액의 구리농도는 대조군과 대상군 사이에 유의한 차이가 없으며 항정신약물은 뇌척수액이나 혈청의 동농도에 영향을 주지 않았다. 둘째, 혈청이나 뇌척수액의 아연농도는 전체대상군과 대조군은 유의한 차이가 없었다. 그러나 대조군과 3주간 위약군의 뇌척수액 아연농도는 유의한 차이가 있었다. 셋째, 항정신약물은 뇌척수액 아연농도에 영향을 주지 않으나 혈청에서는 투약후 아연농도가 유의하게 낮았다. 넷째, 혈청의 동농도와 뇌척수액의 동농도 및 혈청아연농도와 뇌척수액 아연농도 사이에는 상관관계는 없었다. (본 연구에 많은 도움을 주신 서울의대 생화학교실원 오석배 용인정신병원장님을 비롯한 용인정신병원직원 여러분께 감사드립니다.) To investigate copper and zinc concentrations in schizophrenia, we have measured copper and zinc concentrations in the serum and CSF of 27 male schizophrenia (mean age 30.2±5.6) and 10 male controls (mean age 42.7±10.6) with atomic absorption spectrophotometry. The results were as follows. 1. There were no significant differences in serum and CSF copper concentrations between groups of controls, unmedicated schizophrenic patients, and schizophrenic patients taking neuroleptics. Neuroleptics did not significantly alter mean copper concentrations in serum and CSF of schizophrenic patients. 2. There were no significant differences in serum and CSF zinc concentrations between groups of controls, and schizophrenic patients. But mean CSF zinc concentrations was lower in a group of 3-week-drug- free schizophrenic patients. 3. Neuroleptics did not significantly alter mean CSF zinc concentrations of schizophrenic patients, but serum zinc concentrations were influenced by neuroleptics. 4. There were no significant correlations between CSF copper (or zinc) concentrations and serum copper (or zinc) concentrations in schizophrenic patients or controls.