PelB Signal Sequence로 유도된 제조합 인간 상피세포 증식인자 분비 발현 벡터의 제조

박세철,남정현,김정근,권태종,고인영,유광현 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5

전사개시 단계중에 삽입되는 N말단의 methionine이 제거되고 pelB signal sequence를 삽입하여 인체 epidermal growth factor (hEGF)가 직접 분비발현 되도록 고안된 T7과 tac promoter에 의하여 각각 발현되는 pYHB101과 pYHB2를 제조하였다. 또한 T7 promoter에 의하여 발현되지만 signal sequence를 삽입하지 않은 pYHB1도 제조하여 각각 제조된 재조합 plasmid들의 발현 및 분비효율을 비교하였다. pelB signal sequence hEGF 유전자를 갖도록 제조된 pYHB101을 포함하고 있는 균주가 pYHB1보다 균체 성장속도는 낮았지만 pYHB101을 1 mM IPTG로 5시간 induction 하였을 때 13 mg/l의 rhEGF가 분비발현되어 T7 promoter의 발현 체계를 갖는 경우가 tac promoter보다 높은 발현 효율을 보였다. 이때 pYHB101에서 발현되는 rhEGF의 발현량은 준최적 조건에서 14 mg/l이었다. We have designed nucleotide sequences of hEGF structural gene to eliminate the N-terminal methionine residue incorporated during the translation initiation step, and constructed recombinant human epidermal growth factor (rhEGF) secretion plasmids pYHB101, and pYHB2 in which pelB signal sequence-hEGF gene was expressed under the control of the T7, and tac promoter, respectively. We also constructed pYHB1 vector which contains rhEGF gene controlled by T7 promoter. The transformant with pYHB101 showed relatively slow growth pattern compared to the transformant with pYHB1. However, we observed that the transformant with pYHB101 secreted rhEGF of 13 mg/l significantly after 5 hr induction with 1 mM IPTG and that the T7 promoter was more effective than tac promoter when connected to pelB signal sequence. The amount of rhEGF was 14 mg/l under the sub-optimized condition.

임신부 뇨로부터 정제된 인간 상피세포 증식 인자 유사체의 in vitro bioassay 및 특성

박세철,전재현,남정현,권태종,고인영,유광현 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.4

벤조산흡착, 음이온 교환수지, 단일클론항체을 이용한 immunoaffinity chromatography를 통하여 임신부 뇨로부터 천연의 hEGF를 정제하였다. 정제된 hEGF는 μ Bonda C_18 column을 사용한 HPLC 분석을 통하여 4개의 fraction으로 분리가 가능하였으며 western blot과 double immunodiffusion 실험 결과, 각각의 fraction이 hEGF의 특성을 가진 유사체인 것을 알 수 있었다. 또한 hEGF 표준 물질과의 spiking 및 아미노산 분석 등을 통하여 두 번째 fraction이 nhEGF와 동일한 것으로 확인하였다. nhEGF 및 그 유사체의 생물학적 활성 비교를 위하여 NIH 3T3 세포주에서 5'-Brdu incorporation 측정을 위한 labelling 시간, 혈청 농도의 최적 조건을 결정하였다. NIH 3T3 세포주의 DNA 합성능은 0.2% FCS가 포함된 저혈청 배지에서 hEGF가 0.1~10 ng/ml 농도로 첨가하였을 때 증가하는 경향을 나타냈다. HPLC를 통하여 분리된 두번째 hEGF 유사체가 다른 유사체보다도 생물학적인 활성이 우수하였으며, rhEGF 표준물질과의 spiking 및 아미노산 서열 분석등을 통하여 nhEGF로 밝혀졌다. 임신부의 뇨의 hEGF 유사체 함량중 natural hEGF는 46%이었다. Natural human epidermal growth facto (nhEGF) was purified from pregnant human urine by benzoic acid adsorption, DEAE-Sepharose ion exchange, and immunoaffinity chromatography. The purified nhEGF was further separated into four fractions using Bondapak C_18 HPLC system. Following characterization by Western blot and double immunodiffusion, we found that each fraction corresponds to four derivatives of the nhEGF. For biological analysis of nhEGF, we optimized the labeling time and serum concentration for the incorporation of 5-bromo-2'-deoxy-uridine (BrdU), a non-radioactive alternative for [^3H]-thymidine uptake, into NIH 3T3 cells. The DNA synthesis of NIH 3T3 cells was gradually increased at the nhEGF concentrations between 0.1~10 ng/ml in the Dulbecco's Modified Eagles Medium (DMEM) containing 0.2% Fetal calf serum (FCS). When we assayed the biological activity of four fractions, the activity of the second fraction was superior to that of the others.

재발 또는 불응성 비호즈킨 림프종 환자에서 CDME 구제항암화학요법 후 고용량 항암화학요법 및 자가말초혈액 조혈모세포이식의 효과

김세형,한강원,배상병,김찬규,이남수,이규택,박성규,원종호,홍대식,박희숙 순천향의학연구소 2004 Journal of Soonchunhyang Medical Science Vol.10 No.1

Background and objectives : The long-term survival in patients with non-Hodgkin's lymphoma (NHL) after conventional dose chemotherapy is about 35% and the rest of the patients tend to have relapse. So, in relapsed or refractory NHL, we compared the outcome of patients undergoing high-dose chemotherapy with autologous peripheral blood stem cell transplantation(APBSCT) with only salvage chemotherapy of cisplatin, dexamethasone, mitoxantrone, and etoposide(CDME). Materials and methods : From June 1993 to December 1999, 25 patients with relapsed or resistant NHL were treated with CDME regimen as salvage chemotherapy. Twelve patients were received four cycles of CDME chemotherapy, and 13 patients were received high-dose chemotherapy with APBSCT following two cycles of CDME chemotherapy. Results : The median follow-up duration was 12.8 months(range:4-68). The overall response rate was 41.7% (complete response rate 25%, partial response rate 16.7%) in 12 patients with CDME only. Thirteen patients who were treated with high-dose chemotherapy with APBSCT achieved 61.5% complete response rate and 15.4% partial response rate, with an overall response rate of 76.9%. The estimated 3-year progression-free survival rate was significantly higher among patients who received high-dose therapy than patients who received CDME only(41.5% vs 20.0%, p<0.05). And, 3-year overall survival rate was significantly higher among patients who received high-dose therapy(51.3% vs 25.0%, p <0.05). Conclusions : In relapsed or refractory NHL, CDME chemotherapy is an effective salvage chemotherapy and allow peripheral blood stem cell collection. Also, high-dose chemotherapy with APBSCT following CDME is superior to CDME salvage chemotherapy only.

강화 특화작목 지표물질의 함량 분석

유종수,방면호,정해곤,정태숙,최명숙,이경태,김세영,백남인 경희대학교식량자원개발연구소 2007 硏究論文集 Vol.26 No.1

강화 특화작목의 건강기능성 식품 제품화의 가능성을 타진하고 재배작물의 표준화 연구를 위해 사자발쑥, 싸주아리쑥 그리고 순무에 대해 총 폴리페놀과 총 플라보노이드 함량을 측정하였다. 총 폴리페놀 함량은 사자발쑥이 9.5mg/g, 싸주아리쑥이 7.4mg/g, 순무가 1.8mg/g으로 총 플라보노이드 함량은 사자발쑥이 4.3mg/g, 싸주아리쑥이 3.6mg/g, 순무가 0.4mg/g으로 확인 되었다. 또한 강화 약쑥의 유효지표성분인 eupatilin과 jaceosidin이 사자발쑥에는 각각 208mg/100g, 118mg/100g, 싸주아리쑥에는 199mg/100g, 111mg/100g 함유된 것을 HPLC를 통해 확인하였다. This study was conducted to investigate the contents of total polyphenol and total flavonoid of three Ganghwa indigenous crops, two varieties of Artemisia princeps Pampanini (Sajabalssuk, Sajuarissuk) and Brassica campestris ssp rapa. Total polyphenol and total flavonoid contents of Sajabalssuk were 9.5 and 4.3 mg/g, those of Sajuarissuk were 7.4 and 3.6 mg/g, those of B. campestris ssp rapa were 1.8 and 0.4 mg/g, respectively. The contents of eupatilin and jaceosidin in A. princeps Pampanini which were calculated by HPLC, were determined 208 mg/100g, 118 mg/100g in Sajabalssuk and 199 mg/100g, 111 mg/100g in Sajuarissuk, respectively.

Diethylstilbestrol계가 노랑초파리의 발육에 미치는 영향

김택수,김종세,조남기 조선대학교 부설 유전생물공학연구소 1991 유전생물공학연구지 Vol.1 No.1

Four strains of Drosophila melanogaster : one wild strain gathered from the native inhabitant group on Ssangchon-dong in Kwangju, Korea, from May 10 to 23, 1988 and three strains bred and kept in incubators of Kwangju Teachers college for many years-namely, Oregon-R, cinnarbar brown and SD-72 were taken for this experiment to find the effects of two medicines are so constitued Diethylstilbestrol, Euvestrol, and Stilbestrol (Jungwon pharm. Co.) respectively, on the development and eclosion of Drosophila melanogaster. Eggs and imagines of these strains were given Euvestrol and Stilbestrol, and bred in incubators separately, and the following results were obtained from observation. When the laid eggs of Drosophila melanogaster were given Euvestrol Inj., no difference was found by strains, but there was difference by quantity of medicines given. When the imagines of Drosophila melanogaster were given Euvestrol Inj., no difference was observed by strains, but some difference by concentration of medicines with serious effect in high concentration. When the laid eggs of Drosophila melanogaster were given Stilvestrol tab., difference of effects was observed neither of strains by quantity of medicines. When the imagines of Drosophila melanogaster given Stilbestrol tab., showed little difference of effects among the four strains, and no difference by quantity of medicines

立地特性別 Asbestos의 環境濃度

유영식,김종규,민달기,김남천,이형춘,최한영,김을환,우세홍,이상건 서울保健大學 1989 論文集 Vol.9 No.1

나환자에 있어서 체액항체에 관한 고찰

김주덕,김세종,심우남 大韓癩學會 1980 癩學 會誌 Vol.13 No.1

시차주사열량분석기를 이용한 치과용 접착시멘트의 경화기전 및 반응열에 관한 연구

김철위,이용근,윤태호,김효종,남세진 大韓齒科器材學會 2000 대한치과재료학회지 Vol.27 No.3

The working time of dental cement has a close relation with the setting time, where the setting reaction can be said as the result of the chemical reaction between cement liquid and powder. The purpose of this experiment was to investigate the setting characteristics and setting rate of dental cements by measuring the exothermic heat(Joule/gram) from the setting reaction at specified times using a differential scanning calorimeter (Differential Scanning Calorimeter, DSC 204, TASC 414/3A Controller, Netzche, Germany). Graphs of time-exothermic heat at 37℃ isothermal condition, showing the setting reaction of dental cements were plotted. Three types of luting cements studied were zinc-phosphate cement (ZPC), polycarboxylate cement (PCC), and glass ionomer cement (GIC). The effects of powder-liquid ratio on the setting characteristics of cements were also included. The amount of heat (calory) released per one gram of cement was calculated at each specified time, and was analyzed. ANOVA and Scheff's multiple comparison test (p=0.05) were used for statistical analysis using SPSS/PC+. The setting rate of cement was not constant during the setting reaction, and different setting characteristics were observed depending on the type of cement and powder/liquid ratio. Generally, 70∼90% of the setting reaction, measured by the exothermic heat release, occurred within 5 minutes after mixing. The exothermic heat released by ZPC was very high immediately after mixing, and the duration of the heat release was prolonged compared with other cements. The exothermic heat release of PCC finished most rapidly. It was generally observed that the amount of heat release (rise in temperature) per one gram of cement was highest for ZPC, followed by GIC, and then PCC. However, heat release during the first 2∼5 minutes after mixing was highest for GIC, followed by ZPC and then PCC. According to the Scheff's multiple comparison test, the duration of the total setting reaction did not show any significant difference among the cements(p>0.05), and also among the different power/liquid ratio groups(p>0.05). It was observed that more heat was released with the lower powder/liquid ratio than the higher powder/liquid ratio.

완전 고환성 여성화 증후군 1례

이원주,김택훈,권경익,김종인,조남규,박세출 啓明大學校 醫科大學 1993 계명의대학술지 Vol.12 No.2

저자들은 외형상 여성으로써 원발성 무월경과 양측 외음부 종양 촉지를 주소로 입원한 complete testicular feminization syncrome 1례를 경험하였기에 이를 문헌고찰과 함께 보고하는 바이다 The 46, XY individual with complete testicular feminization syndrome is a phenotypic female who is usually somewhat tall, has excellent breast development and presents to the gynecologist because of primary amenorrhea and an inguinal mass. In complete testicular feminization syndrome, axillary and pubic hair are sparse or absent and some degree of virilization such as clitorial hypertrophy, partial labioscrotal fusion and hirsutism doesn't occur at puberty. We report a case of complete testicular feminization syndrome with the brief review of literatures.

순무(Brassica campestris ssp. rapa) 뿌리로부터 지질화합물의 분리 및 동정

방면호,오영준,유종수,한민우,정해곤,정태숙,최명숙,이경태,김세영,백남인 경희대학교식량자원개발연구소 2007 硏究論文集 Vol.26 No.1

순무 뿌리로부터 활성 물질을 분리 동정 하기 위하여 80% MeOH 수용액으로 추출하고 이를 여과, 감압 농축하여 MeOH추출물을 얻었다. 이를 EtOAc분획, n-BuOH분획, H_(2)O분획으로 나누었으며, EtOAc분획에 대해 silica gel column chromatography를 실시하여 3종의 지질화합물을 분리 정제하였다. ^(1)H-NMR, ^(13)C-NMR, DEPT spectrum 및 Mass spectrum등을 통하여 palmitic acid methyl ester(compound 1), linolenic acid methyl ester(compound 2), linoleic acid methyl ester(compound 3)으로 구조를 결정하였다. In order to isolate and identify the active compound from the Brassica campestris ssp rapa, the roots and the root skin were extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH and H_(2)O. From the EtOAc fraction, three compounds were isolated through the repeated silica gel column chromatographies. From the result of spectral data including NMR and MS, the chemical structures of the compound were determined as palmitic acid methyl ester (compound 1), linolenic acid methyl ester (compound 2), linoleic acid methyl ester (compound 3).