Efficient Interleukin-21 Production by Optimization of Codon and Signal Peptide in Chinese Hamster Ovarian Cells

( Hee Jun Cho ),( Byung Moo Oh ),( Jong-tae Kim ),( Jeewon Lim ),( Sang Yoon Park ),( Yo Sep Hwang ),( Kyoung Eun Baek ),( Bo-yeon Kim ),( Inpyo Choi ),( Hee Gu Lee ) 한국미생물 · 생명공학회 2019 Journal of microbiology and biotechnology Vol.29 No.2

Interleukin-21 is a common γ-chain cytokine that controls the immune responses of B cells, T cells, and natural killer cells. Targeting IL-21 to strengthen the immune system is promising for the development of vaccines as well as anti-infection and anti-tumor therapies. However, the practical application of IL-21 is limited by the high production cost. In this study, we improved IL-21 production by codon optimization and selection of appropriate signal peptide in CHO-K1 cells. Codon-optimized or non-optimized human IL-21 was stably transfected into CHO-K1 cells. IL-21 expression was 10-fold higher for codon-optimized than non-optimized IL-21. We fused five different signal peptides to codon-optimized mature IL-21 and evaluated their effect on IL-21 production. The best result (a 3-fold increase) was obtained using a signal peptide derived from human azurocidin. Furthermore, codon-optimized IL-21 containing the azurocidin signal peptide promoted IFN-γ secretion and STAT3 phosphorylation in NK-92 cells similar to codon-optimized IL-21 containing original signal peptide. Collectively, these results indicate that codon optimization and azurocidin signal peptides provide an efficient approach for the high-level production of IL-21 as a biopharmaceutical.

β-Carotene과 알코올을 섭취한 쥐의 혈액 성분변화의 상관관계

고재웅,조만희,이상한,우기민,염윤기,김창세 순천향의학연구소 2000 Journal of Soonchunhyang Medical Science Vol.6 No.2

The study was carried out to investigate the effects of dietary carotenids and β-carotene on the hematological changes caused by alcohol administration into the rats. The results were analyzed using several statistical methods; SPSS computer program for measuring the mean values and standard deviations, one way ANOVA for the test of significance, Duncan method for post HOC multiple comparison, and Pearson method for measuring the correlation coefficient. The following are the conclusion from those statistical analysis. 1. ALB and TB were shown to have the most significant correlation among the blood compositions f개m the basic dietary groups. 2. BUN and CHO were shown to have the most positive correlation among those from the carrot-fed groups. 3. CA and AG were shown positively correlated, and CHO and IP negatively correlated, respectively among those from the cheese-fed groups. 4. AP was shown positively correlated among those from the alcohol-administrated groups. 5. CHO, CA and BUN were shown positively correlated, and CA was negatively correlated among those from the β-Carotene -fed groups. 6. TB and TP were not affected by the correlations between the blood composition from all the experimental groups.

정신분열병에 대한 리스페리돈의 효과 및 안정성

이민수,김용구,김영훈,연병길,오병훈,윤도준,윤진상,이철,정희연,강병조,김광수,김동언,김명정,김상훈,김희철,나철,노승호,민경준,박기창,박두병,백기청,백인호,손봉기,손진욱,양병환,양창국,우행원,이정호,이종범,이홍식,임기영,전태연,정영조,정영철,정인과,정인원,지익성,채정호,한상익,한선호,한진희,서광윤 大韓神經精神醫學會 1998 신경정신의학 Vol.37 No.1

연구목적 : 본 시험의 목적은 임상시험 시작전에 연구자들을 대상으로 PANSS Workshop을 통하여 PANSS, ESRS에 대한 국내에서의 표준화 작업을 구축하고 새로운 정신병 치료제인 리스페리돈의 효과와 안정성을 재확인하여 리스페리돈 사용에 대한 적정화를 이루는데 있다. 연구방법 : 1996년 4월부터 1996년 9월까지 국내 39개 대학병원 정신과에 입원중인 혹은 증상이 악화되어 입원하는 정신분열병 환자 377명을 대상으로 다시설 개방 연구를 시행하였다. 1주일간의 약물 배설기간을 가진후, 리스페리돈을 8주간 투여하였고, 기준점, 1주, 2주, 4주, 그리고 8주후에 평가되었다. 용량은 제1일에는 리스페리돈 1mg씩 1일 2회, 제2일에는 2mg씩 1일 2회, 제3∼7일에는 3mg씩 1일 2회 투여하였다. 이후 환자의 임상상태에 따라 임의로 증량할 수 있으며, 최대 일일 16mg을 초과하지 않도록 하였다. 추체외로 증상을 조절하기 위한 투약을 허용하였다. 임상증상 및 부작용의 평가는 PANSS(Positive and Negative Syndrome Scale), CGI(Clinical Global Impression) 그리고 ESRS(Extrapyramidal Symptom Rating Scale)을 사용하였다. 연구결과 : 377명중 343명(91%)이 8주간의 연구를 완결하였다. 치료 종결시점인 8주후 PANSS 총점수가 20% 이상 호전된 경우를 약물 반응군으로 정의할때, 약물반응군은 81.3%였다. 리스페리돈에 반응하는 예측인자로는 발병연령, 이전의 입원 횟수, 유병기간이 관련 있었다. 리스페리돈은 1주후부터 PANSS양성, 음성, 및 일반정신병리 점수상에 유의한 호전을 보여 효과가 빨랐다. CGI의 경우도 기준점에 비해 1주후부터 유의한 감소를 나타내었다. ESRS의 경우, 파킨슨 평가점수는 기준점과 비교해 투여 1주, 2주, 4주후 유의하게 증가되었다가 8주후 기준점과 차이가 없었다. Dystonia 평가점수는 1주후만 유의한 증가를 보였으며, dyskinesia 평가점수는 유의한 차이가 없었다. 혈압, 맥박수의 생명징후 및 일반 혈액학 검사, 생화학적 검사, 심전도 검사에서 유의한 변화는 없었다. 결 론 : 이상의 다시설 개방 임상 연구를 통해 리스페리돈은 정신분열병 환자에서 양성증상뿐만 아니라 음성증상 및 전반적인 증상에도 효과적인 것으로 사료된다. 보다 명확한 평가를 위해서는 다른 항정신병약물과의 이중맹검 연구가 필요할 것으로 생각되며, 또한 장기적 치료에 대한 평가도 함께 이루어져야 하겠다. Objective : The purpose of this study was to investigate the efficacy and safety of risperidone in the treatment of Korean schizophrenic patients. Method : This multicenter open study included 377 schizophrenic patients drawn from 39 university hospitals. After a wash-out period of 1 week, the schizophrenic patients were treated with risperidone for 8 weeks and evaluated at 5 points ; at baseline, and 1, 2, 4 and 8 weeks of treatment. The dose was increased from 2mg/day(1mg twice daily) to 6mg/day(3mg twice daily) during the first week and adjusted to a maximum of 16mg/day over the next 7 weeks according to the patient's clinical response. Medication to control extrapyramidal symptoms was permitted. The psychiatric and neurological status of the patients was assessed by PANSS, CGI, and ESRS scales. Results : 343(91%) of 377 patients completed the 8-week trial period. Clinical improvement, as defined by a 20% or more reduction in total PANSS score at end point, was shown by 81.3% of patients. The predictors of response to risperidone were associated older age, shorter duration of illness, fewer previous hospitalization. Risperidone had rapid onset of action ; a significant decrease of the total PANSS and three PANSS factor(positive, negative, general), and CGI was already noticed at the end of first week. For the ESRS, parkinsonism rating scores were significantly increased until week 4 comparing with baseline. Dystonia rating scores were significantly increased until week 1, and dyskinesia rating scores were not significantly changed during the study. Laboratory parameters including vital sign, EKG, hematological, and biochemical values showed no significant changes during the trial. Conclusions : This study suggests that risperidone is generally safe and effective against both the positive and negative symptoms in our group of patients.

구강에 발생한 편평상피암종의 MMP-2, MMP-9, TIMP-1, TIMP-2 발현과 EGFR 및 E-cadherin 발현의 상호연관성

조성재,권창석,김용환,윤혜경,김우형,이희철 인제대학교 백병원 2002 仁濟醫學 Vol.23 No.4

Objective : Matrix metalloproteinases(MMPs) are involved in the degradation of extracellular matrix, which is related to infiltrative growth and metastasis of tumor. Positive correlation between MMPs expression and clinico-pathologic parameters were reported, and MMPs expressions were regulated by tissue inhibitor of metalloproteinases(TIMPs) or cell adhesion molecules such as E-cadherin and epidermal growth factor receptor EGFR). The aim of this study is to evaluate the relationship between MMP-2, MMP-9 expressions and clinico-pathologic factors such as clinical stage. recurrence. histologic grade and invasion pattern, and between MMP-2 and MMP-9 Expressions and TIMP-1, TIMP-2, and E-cadherin expressions. Methods and Material : The materials were 55 cases of squamous cell carcinoma of the oral cavity, and immunohisdtochemical studies for MMP-2, MMP-9, TIMP-1, TIMP-2, E-cadherin and EGFR were performed. Clinico-pathologic factors such as clinical stage, recurrence, histologic grade and invasion pattern were recorded through microscopic examination and clinical record review. Statistical analyses between MMP-2, MMP-9 expressions and clinico-pathologic factors, TIMP-1, TIMP-2 expressions and EGFR and E-cadherin expressions were done. Results : 1. MMP-2 and MMP-9 expressions were noted in 30(54.5%) and 22(40.0%) of 55 cases, respectively. TIMP-1 and TIMP-2 expressions were observed in 21(38.2%) and 33(60.0%), and E-cadherin and EGFR expressions were found in 35(63.6%) and 26(47.3%) of 55 cases, respectively. 2. MMP-2 expression rate was slightly higher in cases without recurrence, and TIMP-2 expression rate was slightly higher in cases showing more infiltrative growth pattern. However, there were no significant differences of MMP-2, MMP-9, TIMP-1, and TIMP-2 expression rates according to other clinico-pathologic parameters were noted. 3. The expression rate of EGFR was higher in cases with well differentiation (p=0.0047), but no positive relationship between the expression rate of E-cadherin and histologic grade was found. There were no significant differences of the expression rates of E-cadherin and EGFR according to other clinico-pathologic parameters. 4. Cases with positive reaction for MMP-9 showed an increasing tendency of negative reaction for TIMP-1. However, signigicant relationships between MMP-2, MMP-9 expressions and TIMP-1, TIMP-2 expressions were not noted. 5. The expression rate of MMP-2 was higher in cases with positive reaction for Ecadherin and EGFR with no statistical significance. The expression rate of MMP-9 was significantly higher in cases with positive reaction for E-cadherin(p=0.0221), but there was no significant relationship between MMP-9 and EGFR expression. Conclusion : These results suggest that MMP-2, MMP-9, TIMP-1 and TIMP-2 expressions are involved in the development of oral squamos cell carcinomas, but MMP-2, MMP-9, TIMP-1, and TIMP-2 expressions might not seem to be a sueful prognostic factors because there were no significant relationship between clinicopathologic parameters, EGFR expression showed positive correlation with low histologic grace, so EGFR expression could be regarded as a good prognostic factor. In the progression of squamous cell carcinoma of oral cavity, MMP-9 and E-cadherin might be involved with positive correlation, but there were no significant correlations between MMP-2, MMP-9 and TIMP-1, TIMP-2 and EGFR expressions rate. Further study for other factors such as cytokines or growth factors except TIMP-1, TIMP-2 and EGFR will be needed.

심환좌심실기능부전을 동반한 만성신부전환자에서 완전폐색관상동맥의 경피적관상동맥확장술

이내희,조윤행 순천향의학연구소 2004 Journal of Soonchunhyang Medical Science Vol.10 No.2

..

분리 유채단백의 전기영동 패턴에 미치는 Phytate의 영향

조희경,윤재영,이서래 이화여자대학교 생명과학연구소 1992 생명과학연구논문집 Vol.3 No.-

영양 저해인자로 알려진 phytate를 제거한 분리 유채단백(low-phytate rapeseed protein isolate)을 제조하여 유채단백질과 phytate와의 결합양상에 미치는 pH 및 phytate 첨가량의 영향을 알아보기 위하여 SDS-polyacrylamide gel 전기영동법을 수행 하였다. 가용화된 분리 유채단백은 pH2.0과 pH5.0에서 phytate 첨가량이 증가할수록 가용성 단백질 band의 수가 감소하였으며 pH11.5에서는 band의 변화가 없었다. pH2.0에서 대조군의 18개 band중 7개의 band(105.8,52.3,27.3, 34.8, 36.3, 21.3, 18.4 KDa)가 100mg phytate 첨가시 침전되었으며 150mg 첨가시에는 6개의 band (78.8, 46.5, 19.4, 16.8, 11.7, 8.5KDa)가 더 사라졌다. ph5.0에서는 대조군과 비교하여 15개의 band중 4개의 band( 34.8, 21.3, 18.4, 16.8KDa)만이 phytate 첨가시 침전되었다. 따라서 유채단백을 식품소재로 이용하기 위해서는 phytate 함량을 낮춤으로써 그의 기능성을 향상시킬 수 있을 것이다. This study was undertaken to investigate the effect of pH and phytate level on the solubility of the protein due to binding between phytate and low-phytate rapeseed protein isolate by means of SDS-polyacrylamide gel electrophoresis. Results showed that the number of protein bands decreased by the increasing amount of phytate added to the soluble extreact at pH 2.0 and 5.0 whereas there was no change at pH 11.5. Among 18 bands of rapeseed proteins at pH 2.0 seven bands (105.8, 52.3, 37.3,26.3,21.3,18.4 KDa) were removed by precipitation with 100mg phytate addition and six bands(78.8, 46.5, 19.4, 16.8, 11.7, 8.5KDa) further disappeared by 150mg phytate addition. Among 15 bands at pH 5.0, only four bands disappeared by phytate addition. It is suggested that thefunctionality of rapeseed protein isolate can be improved by lowering the phytate content.

창원시의 A하천 유입오염부하량 실태조사

윤조희,이상훈,조성호 경남대학교 환경문제연구소 1998 환경연구 Vol.21 No.-

To obtain basic data for water quality management and improvement of A-stream located at changwon city, this study was carried out to investigate and estimate flowrates and the characteristics of water pollutants such as organics and nutrients etc, discharging into main A stream where located to flow at commercial and residential areas. A stream was divided five section and field studies have been from June 1997 to May 1998 for one year. The results obtained from this research were follows. The flowrates at the lower site(A-5) of A stream were varied from 19,967 ㎥/day in winter to 29.074㎥/day in summer season. And BOD and COD concentrations at the same sampling site were yearly mean values 45.4㎎/L and 21.38㎎/L, and loading rates 1042.7㎏/d and 548.1㎏/d, respectively. The ratio of BOD loading in A-3 to A-5 to total section were occupied 0.90. T-N and T-P concentrations were obseved average values 27.79㎎/L and 1.47 ㎎/L yearly, loading rates 477.04 ㎏/d and 29.75 ㎏/d, respectively. The value for ratio of T-N loading in A-3 to A-5 to total section were 0.86. similar to the above value, 0.90 for BOD loading ratio. It is consequently necessary to take into account that the policy and master plan must firstly be to reduce pollutants loading rates inputing into A-3 to A-5 at A stream.

중수도 처리용 오존발생기 처리성능평가

윤조희 경남대학교 환경문제연구소 1997 환경연구 Vol.19 No.-

In oder to determine the feasibility of using ozone for wastewater reclamation and reuse, effluent treated by an activated sludge process as model water was ozonated in cylinderical bubble column reactor. Pilot-scale plant was operated 1㎥/day under ozone raction time of one hour. The results of this study are as follows. Ozonation could improve the characteristics of model water for reuse in terms of organic material(BOD), color and E.coliform bacteria, but not improve that of water for SS, TN and TP. This study found that ozone process was applicable as a unit, process for wastewater reuse system.

활성 슬러지와 UF막을 연계한 공법에 있어서 FLUX에 미치는 운전인자의 영향

윤조희,서정범 嶺南大學校 環境問題硏究所 1995 環境硏究 Vol.15 No.1

활성슬러지와 UF막을연계조합한 공정을 이용하여 운전조작인자를 변화시켜 처리수의 플럭스에 미치는 영향에 대하여 실험한 결과 다음과 같은 결론을 얻었다. 1)유기물의 용적부하가 증가함에 따라 처리수의 BOD농도는 다소 상승하는 경향이 있었으나 항상 10mg/L이하이었고, SS농도는 전 공정에서 1mg/L 이하로 안정된 수질을 나타내었다. 또한 막면유속은 처리수의 수질에는 영향을 미치지 않았다. 2) 유기물 용적부하 0.4kgBOD/m^3.d에서 막면유속이 0.6m/sec까지는 슬러지 부착현상 등에 의하여 플럭스가 0.05m^3/m^2.d정도로 안정되게 유지되었다. 3) 막면 유속을 1,/sec에서 유기물 용적부하를 0.8kgBOD/m^3.d이하인 경우에는 0.15m^3/m^2.d이상의 높은 플럭스를 나타내었으나 그 이상으로 높게 되면 슬러지부착 및 막의 막힘현상이 발생하여 플럭스가 감소하였다. 4) 반응조내의 미소생물상은 막면유속에 영향을 크게 받았으며 원생동물의 수는 막면유속증가와 함께 지수적으로 감소하였다.

SBR를 이용한 생물학적 탈인에 있어 유기물 및 인 거동에 관한 속도모델

윤조희 경남대학교 환경문제연구소 1996 환경연구 Vol.18 No.-

SBR를 이용하여 혐기/호기법에 의한 생물학적 탈인 과정에 대하여 2종류의 미생물과 인함유량을 바탕으로 속도모델화를 실시하여 인 및 유기물의 농도변화를 수치모사에 의해 추정하여 실험치와 비교검토하고 최적운전조건을 탐색하였다. 1 싸이클동안에 인의 농도변화는 시스템내의 탈인 미생물의 비율(f)와 혐기상태의 초기단계에 있어 유기물의 농도가 인의 거동에 크게 영향을 미치는 것으로 나타났다. 실험결과와 계산결과를 비교하여 본 결과, 제안모델에 의해 양호하게 농도의 변화특성을 나타낼 수 있다는 것이 확인되었다. 생물학적 탈인과정의 최적조건을 탐색한 결과, 유기물의 농도와 탈인성 미생물의 비율, 운전시간의 비율 등에 따라 다른 결과를 나타내었다. 본 시스템에서 탈인율을 80%이상 유지하기 위해서는 총 반응시간중에 혐기시간의 비율을 모든 조건에서 0.4~0.6정도로 설정하여 운전할 필요가 있었다. A kinetic model that describes combination of two species of biomass and their phosphorus content in a biological phosphorus removal system using sequencig batch reactor (SBR) under cyclic anaerobic and aerobic conditions has been developed for process simulation. The values calculated from this model were compared with experimental data. And in seeking to optimize the simulated results for the influent, and effluent concentrations of phosphorus and organic carbon at a number of operating parameters were also used. The changes of liquid phase phosphorus concentrations during a cyclic period was found to depend on the ratios(f) of phosphorus accumulating biomass to total biomass and on the biodegradable organic carbon concentration at initial step of anaerobic period. The prediction of changes of these concentrations in the system calculated by the proposed model gives good agreement with the experimental results. In order to maintain over 80% of phosphorus removal efficiency from these optimized results, this SBR system roust be operated on about 0.4-0.6 ratios of anaerobic to total reaction time under all conditions.