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Escherichia coli(M15)을 이용한 serotonin N-acetyltransferase 유전자의 발현
김용만,정미영,윤경식,진병관,백행운,조용호,백형환 慶熙大學校 1997 論文集 Vol.26 No.-
AA-NAT cDNA was obtained by RT-PCR technique from total RNA of rat sacrified at night(02:00am). pCRNAT was cloned using the pCRII vector with insertion of AA-NAT cDNA(about 1.4 kb) at EcoRI site. For the expression of the gene, pQECNAT was subcloned, in which the coding region of AA-NAT was inserted into expression vector pQE30 at BamHI and HindIII sites. According to the experimental results, Escherichia coli strain M15, transformed by the expession vector pQECNAT, was selected as a host to express AA-NAT gene with the induction of isopropyl β-D-thiogalactoside(IPTG). Optimal condition for the expression of AA-NAT gene was achieve from the experimental results, showing that the expression of the protein was inducible much 19.800 ± 2,200 dpm per ml of culture volume in 4 hours at the concentration of 2 mM IPTG. Partial purification through the affinity column(Ni-NTA agarose) binding to the continuous 6 histidine residues of protein resulted in 5 times more increase in the specific activity of AA-NAT than that of the homogenate of bacterial pellet. These experimental results will provide basic data in further study for the enzymatic kinetics and antibody production of AA-NAT.
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한용진,조봉균,정재훈,이수빈,박해윤,이태기,채희백,홍진수 순천향대학교 기초과학연구소 1998 순천향자연과학연구 논문집 Vol.4 No.2
Thin amorphous carbon films were deposited by a Q-switched Nd;YAG 532nm with beam power density of ?? on the high purity graphite (99.7%). The pressure of vacuum chamber was ?? Torr. In order to estimate the quality of the Pulsed Laser Deposition films one of the most important optical properties, bandgap energy, was characterized by transmission and reflection in the range of the visible, and an optical direct bandgap energy of 2.45eV and indirect bandgap energy 0.36eV were obtained. Surface morphology of amorphous film was investigated by AFM (Atomic Force Microscopy). Its surface roughness is 70nm. Comparing our results with the published values in the literature we have confidence that our films show the good quality for optical properties.
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망막아세포종 세포주(Y-79)의 Arylalkylamine N-acetyltransferase 활성에 미치는 Dopamine의 영향
김석민,정미영,윤경식,진병관,백행운,백형환,조용호 慶熙大學校 1997 論文集 Vol.26 No.-
The arylalkylamine N-acetyltransferase(AA-NAT, EC 2.3.1.87) in vertebral pineal gland and retina is rate-limiting enzyme in the pathway of biosynthesis of melatonin. AA-NAT activities are remarkably increased during night with 10-150 folds. Neurohormone melatonin is thought to play roles in a broad range of physiological functions in human, primarily through effects on the biological clock in the suprachiasmatic nucleus of the hypothalamus, This rhythm reflects large changes in the activities of AA-NAT. So the important regulatory role of AA-NAT has made it central interest in research on melatonin biochemistry and neurochemical signal transduction. The neurotransmitter that regulates the AA-NAT activities is norepinephrine, which acts through a cyclic AMP mechanism. The effect of dopamine on AA-NAT in human retinoblastoma Y-79 cells was studied. the obtained results are as follows; AA-NAT activities in Y-79 cells were increased by cyclic AMP, and dopamine inhibited the AA-NAT activities stimulated by forskolin. The pattern of inhibition of dopamine on the AA-NAT activities in Y-79 cells was time and concentration dependent. The results suggest that dopamine participates in the regulation of the AA-NAT activities through the inhibitory mechanism in the human pineal and retinal tissues.
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Cho, Baik Kee,Jeong, So Hee,Lee, Dong Won,Park, Chul Jong,Lee, Sang Chin,Paik, Seung Churl,Cho, Sang Hyun,Yi, Jong Yuk,Kim, Tae Yoon,Kim, Jin Wou,Kim, Si Yong,Cheong, Ji Yun,Yang, Kyung Mee 대한의진균학회 2000 대한의진균학회지 Vol.5 No.1
이 트라코나졸은 약물역동학적으로 피부 각질에 친화성이 높고 투약 종료 후에도 4주간 지속되며 고용량 단기간 투여가 효과적이라는 것이 알려지면서 각화형의 수장부/족부 백선 치료에도 고용량단기간 투여법이 시도되고 있다. 본 연구는 각화형의 수장부/족부 백선 환자 97명을 대상으로 이트라코나졸 1일 400 mg을 7일간 투여하여 효과 및 안전성을 평가하였다. 투약 전, 투약 후, 투약 종료 4주후에 임상적, 진균학적으로 효과를 평가하여 임상적으로 73.8%에서 중등도 이상의 개선, 진균학적으로는 84.5%의 완치율을 얻었다. 97.6%의 환자에서 양호한 내약성을 나타내었고, 2.4%에서 위장관 증상, 피부 발진 등이 발생하였으나 특별한 치료 없이 호전되었다. 이상과 같은 결과로 각화형의 수장부/족부 백선 치료에 이트라코나졸 400mg 7일 투여는 효과적이면서 동시에 안전한 치료법임을 확인하였다. [의진균지 5(1): 7-12]
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Development of a New Herbal Anti-Arthritis Drug, Joins™(SKI 306X)
Cho, Yong-Baik 경희대학교 2001 INTERNATIONAL SYMPOSIUM ON EAST-WEST MEDICINE Vol.2001 No.1
Chronic arthritis is classified into two types, degenerative and rheumatoid arthritis. Degenerative arthritis called as osteoarthritis, which is most frequently occurring, causes degenerative figures of knee, waist and knuckle, and accompanies severe pain around the cartilage. Also, it may cause morning stiffness, gelling effect, tenderness, bone swelling, crepitus, and motion disorders. Rheumatoid arthritis has three most likely etiological factors such as heretical cause of host, immune regulation disorder (autoimmune disease), and external infection. However, since there are too many varieties in causes and symptoms, it is generally hard to conclude that arthritis is caused by one or two specific factors. Various medicinal herbs have been used for the treatment of paralyzed symptom (痺症, pain, dull sense or insensibility of muscular joint) caused by wind (風), cold (寒), and/or humidity (濕) in East Asia including Korea, China, and Japan. The paralyzed symptom (痺症) is classified into wind pain (風痺), cold pain (寒痺), and humidity pain (濕熱痺) according to records of oriental traditional medicinal books. In these books, wind pain is a symptom that pain is not localized in a part and shifts all around the body. Cold pain has such a symptom that pain is very severe and is localized in one part of the body. On the other hand, the symptoms of humidity pain are such that pain is fixed owl a part and accompanying flare, tumefaction, and the feeling that joint is burning severely. For the treatments of these diseases, anti-inflammatory & detoxificating drug (淸熱藥), sweating & refreshing drug (解表藥), and anti-cold & wined that means anti-germinal drug (去風濕藥) have been used, which could correspond to anti-inflammatory, analgesic, circulation motivating, and antipyretic drug of modern chemotherapies, respectively. To develop a new herbal anti-arthritis drug, SK Chemicals and SK Pharma scientists selected about sixty medicinal herbs by referencing chinese traditional medicinal books and scientific journals that report various pharmacological activities. After investigating these herbs for the anti-inflammatory activity, analgesic effect, blood-circulating improvement, and inhibition activities of articular cartilage break-down enzyme, six medicinal herbs were selected for the further studies. Adjuvant induced arthritis test (chronic arthritis model) and Randall-Selitto's test (anagesic model) were adopted to screen these six candidates. Eventually three medicinal herbs, Clematis mandshurica, Trichosanthes kirilowii, and Prunella vulgaris, were determined as final materials for a new herbal anti-arthritis drug. Later on, JoinsTM (SKI 306X) was developed by investigating optimal mixing ratio of the three herbs (1:2:1, w/w), extraction and partitioned active n-butyl alcohol fraction. JoinsTM showed blood-circulating improvement and modifying effect of stimulated immune systems (no effect on normal immune system) to normal level as well as strong anti-inflammatory and analgesic activities. In addition, inhibiting degradation of joint articular cartilage or accelerating its biosynthesis will be important factors in regulating arthritis. It was reasoned that protecting activity for joint articular cartilage is essential to eliminate fundamental causes of arthritis. We demonstrated excellent joint articular protecting activity of JoinsTM by studying inhibitory activity against rhIL-1α induced proteoglycan degradation in culture system (in vitro) and histologically protecting activity for joint articular cartilage in collagenase induced rabbit arthritis model system (in vivo).Because a long-term medicinal treatment is needed to regulate chronic disease efficiently, we performed toxicity studies and verified that JoinsTM has no noticeable toxicity (LD50 > 5.0 g/ kg, daily maximum tolerance dose >3.0 g/ kg/ day at 4-weeks study). Based on the results from this pre-clinical study, we obtained a provisional registration approval on the condition of clinical trials from KFDA. A double-blind placebo controlled phase II clinical trial was performed for the determination of optimal dosage and evaluation of clinical efficacy. From this study, it was confirmed that administration of JoinsTM by 200 mg t.i.d., was optimal, and also demonstrated that JoinsTM is a rather safe drug with high efficacy. To compare the efficacy and safety profiles of JoinsTM with those of diclofenac in sustained release formulation, which is the most widely used NSAIDs in the world, we performed randomized, double-blind phase III clinical trial at multiple centers including Seoul National University Hospital. From this study, JoinsTM demonstrated that its efficacy was clinically comparable to that of diclofenac, and with respect to safety profile such as the incidence rate of drug-related adverse events, JoinsTM was reported to be about 2 times more tolerable than diclofenac (p=0.017). In case of gastro-intestinal adverse event, known as a typical adverse event of the NSAIDs, JoinsTM showed 3 times more tolerable than diclofenac (p=0.021). From all of these studies, we would like to conclude that JoinsTM is an efficient agent for the arthritis, showing excellent profiles of the safety and efficacy with not only the anti-inflammatory and analgesic activities, but also the biologically effective multi functions including protecting activities for articular cartilage.
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