Cloning of Human Interleukin-2 cDNA in E. coli by Using Oligonucleotide Primers

강성만,김성완,정일엽,나도선,김지영,한문희,Kang, Seong-Man,Kim, Sung-Wan,Chung, Il-Yub,Na, Doe-Sun,Kim, Ji-Young,Han, Moon-Hi 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.1

사람 인터루킨-2(IL-2)의 cDNA 클론을 oligo-누클레오티드를 primer로 사용하여 분리하였다. 사람 leukaemic T-cell line인 Jurkat 세포로부터 mRNA를 분리 하였다. ss-cDNA를 합성하기 위하여 인터루킨-2를 코딩하는 mRNA의 3' 끝쪽에 상보적인 30 mer oligo-누클레오티드를 역전사 반응시 primer로 사용하였으며, ds-cDNA를 합성하기 위해서는 만들어진 ss-cDNA의 3' 끝쪽에 상보적인 oligo-누클레오티드를 primer로 사용하였다. 이 ds-cDNA를 사용하여 partial cDNA library를 만든 뒤 cDNA 합성에 사용한 oligo-누클레오티드를 probe로 사용하여 콜로니 hybridization을 하여 인터루킨-2 cDNA를 찾기위하여 screen하였다. 약 200개의 transformants 중에서 세 클론이 positive signal을 나타냈다. 제한효소지도를 작성하고 누클레오티드 염기서열을 결정함으로써 이들 세 클론이 모두 인터루킨-2 cDNA를 포함하고 있음을 밝혔다. 이 결과는 우리가 만든 partial cDNA library에 인터루킨-2 cDNA가 Taniguchi 등 (1983)이 만든 total cDNA library에 들어있는 것보다 약 300배 가량 증가되어 있음을 시사한다. A cDNA clone for human interleukin-2 (IL-2) was isolated by using oligonucleotides as primers for the first and the second cDNA syntheses. Total RNA was prepared from Jurkat, a human leukaemic T-cell line, cells and mRNA was isolated. To synthesize ss-cDNA, a 30 mer oligonucleotide was used as a primer in the reverse transcriptase reaction. The sequence of the oligonucleotide was complementary to the 3' end of the coding sequence of IL-2. ds-cDNA was synthesized by DNA polymerase reaction using another oligonucleotide as a primer. A partial cDNA library was prepared using the ds-cDNA and screened for the presence of IL-2 cDNA by colony hybridization using the same oligonucleotides that were used in the cDNA synthesis reactions as probes. Three out of 200 transformants showed positive signals. Analysis of these three clones by restriction enzyme mapping and nucleotide sequencing showed that all of them contained IL-2 cDNA. Our results indicated that the IL-2 cDNA was enriched in the partial cDNA library about 300 fold over the population of IL-2 cDNA in the total cDNA library reported by Taniguchi et al. (1983).

Oligo - 누클레오티드 primer 를 이용한 사람 인터루킨 - 2 - cDNA의 E . coli 내 클로닝

강성만,김성완,정일엽,나도선,김지영,한문희 ( Seong Man Kang,Sung Wan Kim,Il Yub Chung,Doe Sun Na ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.1

A cDNA clone for human interleukin-2 (IL-2) was isolated by using oligonucleotides as primers for the first and the second cDNA syntheses. Total RNA was prepared from Jurkat, a human leukaemic T-cell line, cells and mRNA was isolated. To synthesize ss-cDNA, a 30 mer oligonucleotide was used as a primer in the reverse transcriptase reaction. The sequence of the oligonucleotide was complementary to the 3` end of the coding sequence of IL-2. ds-cDNA was synthesized by DNA polymerase reaction using another oligonucleotide as a primer. A partial cDNA library was prepared using the ds-cDNA and screened for the presence of IL-2 cDNA by colony hybridization using the same oligonucleotides that were used in the cDNA synthesis reactions as probes. Three out of 200 transformants showed positive signals. Analysis of these three clones by restriction enzyme mapping and nucleotide sequencing showed that all of them contained IL-2 cDNA. Our results indicated that the IL-2 cDNA was enriched in the partial cDNA library about 300 fold over the population of IL-2 cDNA in the total cDNA library reported by Taniguchi et al. (1983).

Free Paper Session : Upper Gastrointestinal Tract 1 ; Prevalence And Risk Factors For Atrophic Gastritis And Intestinal Metaplasia

( Na Young Kim ),( Dong Ho Lee ),( Joo Sung Kim ),( Hyun Chae Jung ),( In Sung Song ),( Kyung Phil Kang ),( Jung Hoon Lee ),( Jae Il Chung ),( Hyun Cheul Choi ),( Taek Man Nam ),( Sang Hyup Lee ),( Yo 대한소화기학회 2007 SIDDS Vol.9 No.-

Background/Aims: The prevalence of gastric cancer and Helicobacter pylori (Hp) infection is high in Korea. This study was performed to evaluate the prevalence rate of atrophic gastritis (AG) and intestinal metaplasia (IM) and their risk factors in the aspect of Hp virulence factors, environmental and host factors in normal population. Methods: The subjects consisted of 389, 135 H. pylori-negative and 254 H. pylori-positive. AG and IM were scored histologically by the Sydney classification in the antrum and body, respectively. Prevalence rate and bacterial factors such as cagA, vacA m1, m2, and oipA; environmental factors such as smoking, alcohol drinking; host factors such as genetic polymorphisms for IL-IB-511, IL-IRN, TNF-A, IL-10-592, IL-10-819, IL-10-1082, IL-8-251, IL-6-572, GSTP1, and p53 codon 72 were evaluated. Risk factors were calculated by multiple logistic regression analysis. Results: The prevalence rate of AG increased from 25%, 0% in the age of 20s, 45% and 22% in the 40s and 50% and 35% in the over 70s in the antrum and body, respectively (p<0.001). In case of IM it increased from 11.1% and 6.4% in the 30s up to 43% and 43% in over 70s in the antrum and body, respectively, (p<0.001). The positive rates of AG and IM were significantly higher in the Hp-positive than in the Hp-negative subjects. Multivariate analysis showed that the risk factors for AG were Hp infection, age ≥60, cagA and vacA m1 positive. In case of IM the risk factors were Hp infection, age ≥60, smoking, spicy food, occupation (unemployed or non professional vs. professional), IL6-572 G carrier over C/C and IL10-592 C/A vs. A/A. Conclusions: The prevalence rate of AG and IM increased proportional to age. The most risk factor for AG and IM was Hp infection. Bacterial factors were important for AG but environmental and host factors were rather important in case of IM.

저수시 하상상황에 따른 조도계수의 적용에 관한 연구

조성만,유상호,박병선,민일규,민병형 東亞大學校 海洋資源硏究所 2002 硏究論文集 Vol.14 No.1

As the roughness coefficient by existing manning table is used by technician's experience and judgement, it isn't always correct. This study as a try to make quantitative roughness coefficient is to suggest a new application mode computerizing roughness coefficient by surveying the velocity and hydraulic condition of small rivers. The bound of roughness coefficient was 0.027∼0.180 and its values distribute uniformly. This means that the roughness coefficient adapt value Manning table cannot fill up the transformed value when the small rivers are low. The roughness coefficient computation mode in this study with hydraulic radius(R), channel slope(I), average grain diameter(D) as subject factors presents as follows. n = 5.97R^0.671 l^0.452 D^0.059

Characterization of IL-3, IL-5, GM-CSF - induced Eosinophils from Human CD34+ Cord Blood Cells

( Kwang Shin Lee ),( Kang Mo Ahn ),( Seung Yeon Nam ),( Dae Yeul Son ),( Se Chang Han ),( Man Yong Han ),( Sang Il Lee ) 대한 소아알레르기 호흡기학회(구 대한소아알레르기 및 호흡기학회) 1999 소아알레르기 및 호흡기학회지 Vol.9 No.4

두통을 주소증으로 하여 입원한 환자 120예에 대한 임상적 고찰

권경만 ( Kyoung Man Kwon ),김광록 ( Koang Lok Kim ),신윤리 ( Yoon Ri Shin ),박선영 ( Sun Young Park ),박동일 ( Dong Il Park ) 경락경혈학회 2012 Korean Journal of Acupuncture Vol.29 No.4

Objectives: The Purpose of this study is to investigate clinical characteristics with 120 patients who have sufferred from headache and were treated. Methods: We classified 120 patients into several groups by International Headache Society(IHS) classification and evaluated the effects of oriental medical therapy on headache. Results: 1. According to the statistics, women patients were more than men. And 50s has highest number of headache patients. 2. In classification by IHS, tension type headache were highest. 3. 24.2% of patients had entered Department of Internal Medicine, Colleage of Oriental Medicine Dongeui Univ for treatment within six month of onset. 4. According to the statistics, headache of whole part has highest number of headache patients. 5. The most number of associated symptoms were nausea, vomiting, dyspepsia etc. 6. The most number of headache patient`s past history were hypertension. 7. In classification by oriental medical differentiation of symptom and signs, the rate of blood deficiency were highest. 8. 64.2% of patients said they were satisfied with treatment, reporting half the frequency of headache or better from before treatment. Conclusions: In conclusion, the present results suggest that oriental medical therapy has effects on headache.

Streptococcus dysgalactiae로부터 분리된 히알루론산과 황화된 유도체의 구조와 항염증 활성

홍창일(Chang-Il Hong),정의길(Eui-Gil Jung),한국일(Kook-Il Han),김용현(Yong Hyun Kim),이성희(Sung Hee Lee),이홍섭(Hong Sub Lee),한만덕(Man-Deuk Han) 한국생명과학회 2016 생명과학회지 Vol.26 No.5

히알루론산(HA, Hyaluronic acid)은 β-1, 3-N-acetyl glucosamine과 β-1, 4-glucuronic acid가 반복된 선형 폴리머 고분자로서 생물학적 활성 및 생체친화성 특성 때문에 의약 및 약학분야에서 중요한 분자로 여겨지고 있다. 본 연구는 HA을 S. dysgalactiae으로 얻고, 화학적 방법을 통해 황화된 히알루론산(S-HA, Sulfated hyaluronic acid)유도체를 합성하여 그 구조와 항염증 활성을 비교하였다. HA의 생산은 S. dysgalactiae를 5 l 생물반응기를 이용하여 대량 배양하여 수용성 히알루론산(HA-WS, water soluble hyaluronic acid)과 비수용성 히알루론산(HA-WI, water insoluble hyaluronic acid)을 분리 정제하였다. 특히 HA-WI를 황화시켜 황화된 히알루론산(S-HA) 유도체를 합성하였으며, 그 수율은 90%로 나타났다. 합성된 S-HA의 구조를 FT-IR 및 ¹H/<SUP>13</SUP>C-NMR를 통해 S. dysgalactiae 로부터 생산된 표준 HA, HA-WS 및 HA-WI와 비교 분석한 결과, 황으로 치환된 양상을 확인하였다. 또한, S-HA의 항염증 활성을 RAW 264.7 대식세포를 통해 확인한 결과, S-HA는 천연 형태의 HA (HA, HA-WS)보다 nitric oxide (NO)와 COX-2 및 PGE₂ 유전자 발현이 유의하게 낮게 발현되었다. 염증 매개 cytokine인 TNF-α (<80 pg/ml) 및 IL-6 (<100 pg/ml)의 생성도 S-HA가 천연 HA보다 낮은 수준으로 정량되었다. 이 같은 결과에서 황화된 S-HA은 천연 히알루론산보다 용해성이 우수하고 염증관련 사이토카인의 생성 억제를 통해 항염증 효과를 나타내므로 염증치료제, 성형 및 생체 적용 약물전달 소재로 그 활용이 기대된다. Hyaluronic acid (HA) is an important macromolecule in medical and pharmaceutical fields. HA is a natural and linear polymer composed of repeating disaccharide units of β-1, 3-N-acetyl glucosamine and β-1, 4-glucuronic acid. This work aimed to confirm the structural characteristics and anti-inflammatory activities of HA and its chemically sulfated-HA. HA was produced from a fed-batch fermentation process using Streptococcus dysgalactiae in a 5 l bioreactor. HA was isolated water-soluble form (HA-WS) and water-insoluble form (HA-WI) from culture medium, and was obtained chemically sulfated-derivative (S-HA) that resulted in a 90% yield from HA-WI. The structural features of the sulfated- HA (S-HA) were investigated by FT-IR and ¹H-NMR spectroscopy. The FT-IR and NMR patterns revealed the similarity in both the FTIR spectrum as well as NMR spectrum of both reference standard and purified HA from S. dysgalactiae. The anti-inflammatory activities of HA and S-HA were examined on LPS-induced RAW 264.7 cells. S-HA was significantly inhibited production of pro-inflammatory mediators such as nitric oxide (NO) and PGE₂ and the gene levels of iNOS and COX-2, which are responsible for the production of NO and PGE₂, respectively. Furthermore, S-HA also suppressed the overproduction of pro-inflammatory cytokine TNF-α (<80 pg/ml) and IL-6 (<100 pg/ml) compared to that of HA-WI. The present study clearly demonstrates that HA-S exhibits anti-inflammatory activities in RAW 264.7 macrophage cells.

중고령자에 있어서 적절한 운동처방의 연구

이배익,정일호,권만근 한국스포츠리서치 2003 한국 스포츠 리서치 Vol.14 No.4

Recently, the diet of middle aged Korean men is changing. Instead of eating Korean foods exclusively, they are consuming more Western foods which are often unhealthy. In addition, they are exercising less. These factors are leading to a rise in lifestyle related disease, such as diabetes, and dangerous medical conditions like hypertension. As a result, some middle aged men have taken up light exercise, such as hiking, walking, swimming etc. However, they don't know how to set up an exercise program, i.e. frequency of exercise, intensity, duration, etc. A 15 week exercise class was set up 36 middle aged men. Before the class began, a physical fitness and blood test were administered to each subject. The same tests were repeated after the 15th week. Based on the initial results, individual programs were prepared for each participant. A step calorie counter was supplied to measure their exercise activity. The step count results were checked everday, and based on the results, the subjects were divided into three groups for analytical purposes. The firts group, sixteen subjects(44%), averaged a little over 8,000 steps per day. The second group, eight subjects(22%), averaged between 8-10,000 steps per day. The third group, twelve subjects(33%), averaged a little over 10,000 steps per day. After fifteen weeks, the total cholesterol, HDL cholesterol and LDL cholesterol of all subjects showed no significant change. The second and third group showing an especially marked decrease in blood sugar. Also, physical fitness improved for the second and third groups. In conclusion, both groups doing the most exercise showed improvement in their TG and blood sugar results. Blood pressure also decreased for these two groups, which was not true for the first group, also, physical fitness improved in the second and third groups, while the first group showed no significant change. It appears that exceeding a certain minimum level of exercise is required to achieve good results. What would happen if group 1 did the same exercise for longer than is weeks.

입학전형에 따른 임상병리과 학생의 학습적응도

도성탁,신두만,권순일 대구보건전문대학교 2006 대구보건대학 論文集 Vol.26 No.-

This study on the correlation between academic achievement and admission process was performed on a group of students who are majoring clinical pathology at Daegu Health College, Academic achievement of a group of students was evaluated on the basis of their average GPA of whole semesters enrolled at the college, In terms of admission type, Bachelor (Dae-Jol) group showed the best GPA among all groups, followed by General (Il-Ban), Rural(Nong-Oe-Chon) and Special (Teuk-Byul) group in a row. From the view point of admission time, Regular Admission (Joeng-Si) group showed a little bit higher GPA than Early Admission (Soo-Shi) group. When classified by the graduated-high schools, the GPA of the students who are from General high school (In-Moon-Kye) was higher than that of those who graduated Technical high school (Shil-Up-Kye). By the gender, female students exceeded male students in GPA in the first (freshman) and the second (junior) year of the college. However, in the third (senior) year, male students exceeded female students. In a conclusion, in order to select the applicants with better academic capability it may be needed to put more importance on General (Il-Ban) admission by type and Regular Admission (Joeng-Si) by time during the new students admission assessment. In addition it is also recommended to give some preferential treatment to the applicants from General high school (In-Moon-Kye).

Effects of Carthamus tinctorius L. seed extracts on regulation of cellular proliferation, prostaglandin E₂synthesis and plasminogen activator activity in the mouse calvarial bone cells

Lee,Dong-Young,Bae,Man-Jong,Kim,Dong-Il,Lee,Tac-Kyun,Nam,Kyung-Soo,Lee,Dong-Kyu,Kim,Cheorl-Ho 경산대학교 생명자원개발연구소 2000 생명자원과 산업 Vol.4 No.-

Medicinal extracts of Honghwain (Carthamus tinctorius L. seed) (CTE) was tested for whether they could inhibit IL-1β-induced PGE₂productional. Cell viability was not significaltly affected by treatment with the indicated concentration of the extracts. The CTE extracts were shown to havethe inhibitory effects against the synthesis of PGE₂. we also examined the effect of the pretreatment with a various concentrations of the CTE extracts then treated the PGE₂-induction agents. Pretreatment of the CTE extracts for 1 h, which by itself had little effect on cell survival, did not enhance the synthesis of PGE₂. Furthermore, the CTE extracts were shown to have the the protective effects against plasminogen dependent fibrinolysis induced by the bone resorption agents of IL-1β. Pretreatment of the CTE extracts for 1 h did not enhance the plasminogen dependent fibrinolysis. Finally, calcitonin showed the inhibitory activity the IL-1β-stimulated bone resorption in the mouse calvarial bone cells having both of the osteoblast and osteoclast cells. Seemingly, pretreatment of the CTE extracts for 1 h reduced the bone resorption. These results clearly indicated that calcitonin and CTE extracts play key roles in inhibition of the osteoclast-mediated bone resorption, and also indicated that the CTE-water extracts are highly stable and applicable to clinical uses in osteoporosis.