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Molecular Cloning of Xanthine Oxidase Gene from Mouse Liver cDNA Library
Lee, Chu Hee,Nam, Doo H.,Huh, Keun 영남대학교 자원문제연구소 1994 資源問題硏究 Vol.13 No.-
Usirn antisera against boyine xanthine oxidase(E. C.1.1.3.22) raiaed by subcutaneous injection in the back of rabbits, the recomniant phages of mouse liver CDNA library in λgt11 were selected throughout the immunochemical method. To validate the selected clones. the recombinant phages were Iysogenized in E. coli Y1099 and their Iysates were analysed for enzyme activity and immunoreactivity. It was verified that lysates of the Iysogens not only exhibited the enzymatic activity but also bound with anti-(xanthine oxidase) antibody when the gene was induced by isopropyl-β-D-thiogalactopyranoside ( IPTG ) .
An Active Monomeric Form of Bovine Milk Xanthiine Oxidase
Lee, Chu Hee,Nam, Doo Hyun,Huh, Keun 영남대학교 약품개발연구소 1995 영남대학교 약품개발연구소 연구업적집 Vol.5 No.-
Upon gel filtration, the commercial bovine milk xanthine oxidase preparation was fractionated into two preparations showing enzyme activity. Native polacrylamide gel electrophoresis showed that one was in a dimeric form and the other was a monomer having molecular weight of 150 kDa. It was also found that this commercial enzyme existed mostly in an active monomeric form without loss of enzyme activity. The rabbit antisera produced against two enzyme preparations cross-reacted well each other. In SDS-polyacrylarnide gel electrophoresis, however, both enzyme preparations yielded two smaller protein bands below 150 kDa, which appeared to bind with both antisera with high affinity but not to retain enzyme activity. It implies that bovine milk xanthine oxidase can lose its activity when monomeric subunit is further degraded.
Lee, Chu Hee,Bang, Jeong Hee,Nam, Doo Hyun 영남대학교 약품개발연구소 1993 영남대학교 약품개발연구소 연구업적집 Vol.3 No.-
Saccharomyces cerevisiae와 Kluyveromyces fragilis의 세포융합을 통해 얻은 융합효모를 calcium alginate bead에 고정화시킨 후 유당으로부터의 에탄올 생산을 시도하였다. 우선 융합효모의 에탄올 내성을 조사한 결과, 모 효모인 K. fragilis 보다 높은 8.0%에서 최대 내성을 나타내었다. 이렇게 고정화시킨 융합효모를 충전식 반응기에 충전하고 에탄올 생산을 실시하였는데, 10% 유당을 공급하고 30℃에서 운전하였을 때 가장 좋은 에탄올 생산능을 보여주었다. 특히 반응기의 희석률이 0.76hr^(-1) 일 때 13.3 g/l/hr의 높은 에탄올 생산능을 얻을 수 있었다. Yeast cells of a fusant strain constructed by protoplast fusion of Saccharomyces cerevisiae and Kluyveromyces fragilis were immobilized on calcium alginate beads. The increment of the ethanol tolerance of this strain to 8.0%, when compared with the parent K. fragilis, was confirmed. Based on the results from jar fermentation, a packed-bed reactor of the immobilized yeast cells was operated. The optimal performance of the immobilized yeast reactor for ethanol production was achieved when supplying 10% lactose (supplemented 1.0% yeast extract) at a temperature of 30℃. The maximal ethanol productivity was obtained as 13.3 g/l/hr at a dilution rate of 0.76 hr^(-1).
LEE, CHU HEE,YANG, SUN A,RHO, JU WON,LEE, SEUNG YUP,NAM, DOO HYUN 영남대학교 약품개발연구소 1992 영남대학교 약품개발연구소 연구업적집 Vol.2 No.-
The fermentative characteristics in ethanol production from lactose, with increased ethanol tolerance, of a fusant yeast strain constructed by protoplast fusion of Saccharomyces cerevisiae and Kluyveromyces fragilis were studied. The ethanol tolerance of this strain was increased to 8.0%, compared with the parent K. fragilis. During batch ethanol fermentation the optimal cultivation conditions for this fusant yeast were an initial pH of 4.5, a culture temperature 30℃, stirring at 100 rpm without aeration in 10% lactose medium (supplied with 1.0% yeast extract). Using this fusant strain in whey fermentation to ethanol, maximum ethanol production reached 3.41% (w/v) (theoretical yield; 66.7%) after a 48 hour cultivation period.
김경찬,이추희 대한약리학회 2014 The Korean Journal of Physiology & Pharmacology Vol.18 No.1
Lung cancer is still the number one cause of death from cancer worldwide. The clinical effect ofplatinum-based chemotherapy for non-small cell lung cancer is constrained by the resistance to drug. To overcome chemo-resistance, various modified treatment including combination therapy has beenused, but overall survival has not been improved yet. In this study, chemo-resistant lung cancer cells,A549/Cis and H460/Cis, were developed by long-term exposure of cells to cisplatin and the proliferativecapability of these resistant cells was verified to be reduced. We found cytotoxic effect of epigallocatechingallate (EGCG), a major catechin derived from green tea, on both the parental lung cancercells, A549 and H460, and their cisplatin resistant cells, A549/Cis and H460/Cis. ELISA and Westernblot analysis revealed that EGCG was able to increase interlukine-6 (IL-6) production per cell, whereasits downstream effector Signal transducers and activators of transcription 3 (STAT3) phosphorylationwas not changed by EGCG, indicating that IL-6/STAT3 axis is not the critical signaling to be inhibitedby EGCG. We next found that EGCG suppresses the expression of both Axl and Tyro 3 receptor tyrosinekinases at mRNA and protein level, explaining the cytotoxic effect of EGCG on lung cancer cells,especially, regardless of cisplatin resistance. Taken together, these data suggest that EGCG impedesproliferation of lung cancer cells including their chemo-resistant variants through downregulation ofAxl and Tyro 3 expression.
HL-60 세포에서 $TNF-{\alpha}$에 의한 MCP-1 발현에 미치는 Betulinic Acid의 효과
김경찬,이추희,Kim, Kyung-Chan,Lee, Chu-Hee 대한약학회 2008 약학회지 Vol.52 No.1
Betulinic acid, a naturally occurring pentacyclic triterpenoid, is found in abundance in the outer bark of white birch (Betula alba). In this study, we investigated if betulinic acid affects cytokine expression from activated macrophage cells. ELISA result showed that stimulation of HL-60 cells with proinflammatory cytokine such as $TNF-{\alpha}$ resulted in MCP-1 release into culture medium. In addition, transcriptional upregulation of MCP-1 in response to $TNF-{\alpha}$ was observed by RT-PCR analysis. However, incubation of HL-60 cells with betulinic acid prior to $TNF-{\alpha}$ treatment abrogated MCP-1 expression in transcription and translational level. Consistent with a number of studies which reported requirement of ERK activation for $TNF-{\alpha}$ expression, Western blot analysis showed that $TNF-{\alpha}-induced$ ERK activation was suppressed by pretreatment of HL-60 cells with betulinic acid. Taken together, our data indicate that betulinic acid exerts its anti-inflammatory effect through inhibition of $TNF-{\alpha}-induced$ ERK activation which is required for the subsequent MCP-1 release.
신경옥,전정례,이추희,김순동,유연수,남두현,이지선,김종연 한국생물공학회 2006 Biotechnology and Bioprocess Engineering Vol.11 No.3
To develop a health-aid preparation of Chinese yam (Dioscorea batatas Decne), lactic acid fermentation was attempted using a mixed starter comprising of Lactobacillus acidophilus, Streptococcus thermophilus, and Bifidobacterium bifidus. The anaerobic fermentation of a 5% Chinese yam flour suspension gave a uniform suspension of pH 4.35, containing 7.76 × 106 CFU/mL lactic acid bacteria (LAB), and which was found to be acceptable to the panel from a sensory assessment. During the administration of the lactic acid fermented (LAF) Chinese yam to Sprague Dawley rats for 6 weeks, a smaller body weight gain, but greater excretion of feces were observed, implying the creation of a healthy gastrointestine on the administration of LAF Chinese yam, which was also confirmed by the gastrointestinal motility of the feed in rats fed on LAF Chinese yam. The constipation induced by loperamide was further suppressed in a rat group fed on a LAF Chinese yam diet, which was qualified from healthy gastrointestinal flora established by LAB. A serochemical analysis revealed a slight improvement in the blood glucose, neutral lipid and total cholesterol concentrations on administration of LAF Chinese yam, suggesting LAF Chinese yam could be served as a healthy-aid preparation, even for hyperglycemia or hyperlipidemia patients.
전정례,이지선,이추희,김종연,김순동,남두현 대한약학회 2006 Archives of Pharmacal Research Vol.29 No.5
In this study, a 40% ethanol extract of Chinese yam flour (Dioscoreae rhizoma), containing 177±58 µg/mL of dioscin, was tested in order to evaluate its pharmacological effects on the gastrointestinal tracts of Sprague-Dawley rats. Via the ingestion of the Chinese yam extract, the secretion of gastric acid was suppressed in the rats, and gastrointestinal motility increased by as much as 10%. The fecal quantity of rats fed on the Chinese yam extract also increased, by more than 40% as compared with that of the controls. The Chinese yam extract was found not to affect the growth of normal intestinal bacteria. However, a great deal of lactose-fermenting bacteria was observed in the fecal samples of rats fed for 6 weeks on 2% Chinese yam extract. This finding would appear to suggest that Chinese yam extract not only induces an improvement in digestive capability, but also affects the conversion of some intestinal flora to helpful bacteria. Our serochemical analyses indicated that serum glucose, neutral lipid, and total cholesterol levels were reduced to some degree by long-term feeding on Chinese yam extract. This finding bolsters the notion that Chinese yam extract may prove helpful as a digestion- aiding agent for patients suffering from hyperglycemia or hyperlipidemia.