류마티스 관절염 환자의 T세포 수용체 Vβ 유전자 레퍼토리 분석

정성수 ( Sung Soo Jung ),황관표 ( Kwan Pyo Hong ),김동욱 ( Dong Yook Kim ),김태환 ( Tae Hwan Kim ),이인홍 ( In Hong Lee ),전재범 ( Jae Bum Jun ),배상철 ( Sang Cheol Bae ),유대현 ( Dae Hyun Yoo ),김성윤 ( Seong Yoon Kim ),이은영 ( 대한류마티스학회 1996 대한류마티스학회지 Vol.3 No.1

목적: 류마티스 관절염(rheumatoid arthritis; RA)의 병인에 있어 중추적인 역할을 하는 것으로 알려진 T세포에 관한 연구가 최근에 관심의 초점이 되고 있으나, 현재까지 한국인을 대항으로 시행된 보고는 없다. 한국인 류마티스 관절염 환자의 관절병변부위에서 T세포 수용체(TCR)의 Vβ유전자 사용의 빈도를 검색하여 한국인 류마티스 관절염 환자에서 사용되는 빈도가 높은 유전자계를 찾는데 목적을 두었다. 방법: 이에 저자는 T세포에 관한 연구의 일환으로 류마티스 관절염 환자 3명과 정상인 4명을 대상으로 말초 혈액과 활액 T세포로부터 추출한 RNA를 이용하여 cDNA를 합성한뒤 cDNA를 주형을 Vβ family specific oligonucleotide를 시발체(primer)로하여 반정량적 역전사 연쇄중합반응(semiquantative RT-PCR)을 시행하여 T세포 수용체(T cell receptor; TCR) Vβ 레퍼토리를 분석하여 다음과 같은 결과를 얻었다. 결과: 1. 정상인 4명의 말초혈액에서 T세포 수용체 Vβ유전자 평균사용빈도는 Vβ7(8.68±3.20%), Vβ3(7.83±2.03%), Vβ(6.74±1.43%)의 순서로 빈도가 높았으며, HLA-DR4 양성군(3명)에서는 Vβ8(7.39±1.71%), Vβ2(7.31±2.30%), Vβ1(7.22±1.54%)의 순서로 빈도가 높았고, HLA-DR4 음성(1명)에서는 Vβ3(17.79%), Vβ4(11.41%), Vβ24(9.8%)의 순서로 사용빈도가 높았다. 2. 류마티스 관절염환자 3명의 말초 혈액에서는 Vβ16(6.90±1.81%), Vβ18(6.89±0.80%), Vβ14(6.58±0.65%)의 순서로 빈도가 높았다. HLA-DR4 양성 환자군에서는 Vβ16(7.52±2.06%), Vβ14(6.96±0.04%), Vβ22(6.78±0.18%)의 순서로 빈도가 높았다. 각각 환자의 결과를 보면 첫번째 환자(HLA-DR4 양성)는 Vβ16(8.98%), Vβ14(6.99%), Vβ22(6.91%)의 순서로 빈도가 높았고, 두번째 환자(HLA-DR13 양성)는 Vβ18(7.79%), Vβ20(5.98%), Vβ24(5.90%)의 순서로 빈도가 높았다. 이를 종합해 보면 류마티스 관절염 환자의 말초혈액에서 Vβ16, Vβ20, Vβ14, Vβ18유전자계가 제한되어 사용되었다. 3. 류마티스 관절염환자의 활액의 T세포에서 Vβ유전자 평균발현빈도는 Vβ20(7.39±1.77%), Vβ18(5.60±1.31%), Vβ12(5.56±1.77%)순으로 높았으며, HLA-DR4양성인 환자군에서는 Vβ12(6.56±0.58%), Vβ20(6.44±0.94%), Vβ4(5.30±0.49%)순으로 빈도가 높았으며, HLA-DR4음성 환자에서는 Vβ20(9.29%), Vβ19(7.57%), Vβ18(7.07%)의 순서로 빈도가 높았다. 각각 환자의 결과를 보면 첫번째 환자(HLA-DR4 양성)는 Vβ12(6.15%), Vβ20(5.77%), Vβ7(5.74%)의 순서로 빈도가 높았고, 두번째 환자(HLA-DR4 양성)는 Vβ20(7.10%), Vβ12(6.98%), Vβ24(5.83%)의 순서이었고, 세번째 환자(HLA-DR13 양성)는 Vβ20(9.29%), Vβ19(7.57%), Vβ18(7.06%)의 순서로 빈도가 높았다. 활액 T세포에서는 Vβ20, Vβ12유전자계가 제한되어 사용되었다. 4. 말초 혈액에 비해 활액 T세포에서 TCR Vβ유전자 사용빈도가 1.5배 이상인 경우는 Vβ5.2, Vβ9, Vβ23 이었으나, 이들 유전자계가 활액 T세포 전체에서 차지하는 사용빈도에 있어서는 각각 Vβ5.2는 1.07±1.26%, Vβ9는 2.17±1.42%, Vβ23는 3.84+1.97%로 낮은 수치를 나타내었다. 각각 환자에서 비교했을때에 첫번째 환자(HLA-DR4 양성)는 Vβ23(3.69%:1.12%), Vβ5.1(3.51%:1.40%), Vβ12(6.15%:3.29%), Vβ6(3.35%:2.10%), Vβ2(5.74%:3.73%)의 유전자계에서, 두번째 환자(HLA-DR4 양성)는 Vβ9(3.58%:0.74%), Vβ19(4.67%:1.52%), Vβ5.2(2.52%:0.87%), Vβ10(4.25%:1.75%), Vβ1(5.74%:2.35%)의 유전자계에세, 세번째 환자(HLA-DR13 양성)는 Vβ6(2.68%:0.15%) Vβ9(2.17%:0.73%), Vβ11(2.30%:1.10%), Vβ20(9.29%:5.98%)의 유전자계에서 활액에서 말초혈액보다 1.5배이상 증가되어 사용됨을 볼수 있었다. 즉 활액에서 말초혈액보다 의미 있게 편향되어 사용되는 Vβ 유전자계는 각각 환자마다 다른 결과를 나타내었다. 5. 류마티스 관절염환자의 말초 혈액과 정상인의 말초 혈액의 T세포의 Vβ 유전자 사용빈도를 비교했을때 류마티스 관절염 환자의 말초혈액 T세포에서 Vβ16(6.90±1.81%:3.23±1.21%), Vβ18(6.89±0.80%:2.72±0.70%) 유전자계가 2배 이상으로 빈도가 높았고, HLA-DR4양성군만 비교하였을때도 Vβ16(7.52±2.06%:3.69±1.46%), Vβ18(6.45±0.28%:2.61±0.86%) 유전자계가 2배 이상 사용이 많았다. 결론: 이상의 성적으로 미루어 보아 류마티스 관절염 각각 환자마다 다른 Vβ 유전자계의 제한적 사용과 편향됨을 볼 수 있었으나, 공통된 유전자계의 증식은 볼 수 없었다. 이러한 결과는 다른 연구보고와는 다른 결과를 보였고, 이것은 대상 환자의 질병의 유병기간이 다르고, 또 유전적인 배경, 생존 환경, 적용된 방법의 차이로 기인한다고 하는 기존의 보고와 부합된다고 사료되는데 질병의 진행단계에 따라 주로 면역반응을 일으키는 항원의 에피토프가 달라짐에 따라 여기에 대항하는 T세포들의 수용체도 이들 에피토프에 반응할 수 있는 수용체를 가진 코론들이 증식하게 된다는 epitope spreading theory에 부합되며, 이는 아주 초기에 질병을 시작하게 유도하는 항원의 특성을 규명하기 위해서는 환자선택이 연구결과에 결정적인 역할을 할 것으로 사료된다. 또 다른 가능성은 각각 환자마다 다른 Vβ유전자계의 증식된 클론들의 CDR3 연기 서열을 규명하여 비록 유전자 서열이 다를 지라도 항원과 결합하는 같은 성상을 갖는 아미노산 motif를 가질 가능성에 대해서도 연구가 필요하리라 사료된다. Objectives: Polymerase chain reaction (PCR) technology was eamine synovial fluid and peripheral T cells in patients with rheumatoid arthritis (RA) to determine the preferential usage of the T cell receptor(TCR) variable region (V) gene. Methods: Oligonucleotide primers specific for individual TCR Vβ gene families were used to amplify the TCR gene products in a semiquantitative assay of their relative utilization in unselected T cell populations. Results: The result of Vβ utilization was generally heterogenous, similar with previous reports. However, the mean expression of Vβ16 and Vβ18 in RA was more preferentially utilized compared to normal donors. The usage of Vβ in peripheral blood from 3 patients with RA demonstrated restrictions in Vβ16, Vβ20 and Vβ18 genes, respectively. Analyses of synovial fluid resulted in restriction in β12, Vβ20 and Vβ20, respectively. Although there was no significant pattern of skewed Vβ gene mean usage when comparing the synovial fluids with the peripheral blood T cells from RA patients, there were significant biased Vβ genes, Vβ12, Vβl and Vβ20, each 3 patients. As the HLA type is a determining factor in shaping TCR repertoire of peripheral T cells, we compared the Vβ utilization in HLA-DR4 expressing groups that have susceptibility and gene dosage effect in disease progression. It was a little different that comparing the pattern of Vβ usage in peripheral blood and synovial fluid from RA patients between HLA-DR4 positive and negative group. Conclusion: The results were consistent with the conclusion that the increased Vβ family T cells infiltrate synovium and are dependent on each patient and may be involved in inducing and maintaining the synovitis that characterizes RA The different outcome of each patient may be due to the difference in disease duration, genetic background and geographic region. A more important factor may be the stage of disease, because epitope induced immune reaction may change over time. Therefore, selecting patients early in the course of disease may be important and may facilitate the need for more in-depth TCR analysis in the future.

소아에서 발생한 골수이형성 증후군 2례

홍성진,윤석중,김성우,조형구,오명호,김기혁,이인성 건국대학교 의과학연구소 1996 건국의과학학술지 Vol.5 No.-

Myelodysplastic syndrome(MDS) is a group of acquired conditions characterized by progressive bone marrow failure associated with normocellular or hypercellular bone marrow that cannot be attributed to nutritional deficiency, chronic infection, or other chronic systemic illness, and cannot be reversed by the successful treatment of those conditions. Whereas the cases of secondary MDS occur at all ages, primary MDS is essentially a disease of elderly, though no age group is exempt and predomonant in men. The MDS is fundamentally clonal disorders at the level of the haemopoietic totipotent stem cell. Clinically, they are characterized by varing degrees of peripheral cytopenia(s) with morphological and functional abnormalities of blood elements. We have experienced two unrelated pediatric cases, who showed pancytopenia on admission and the characteristic marrow findings of MDS. The first case, a 12 months-old girl, showed increased reticulin with abnormal localization of immature precursor cells(ALIP) in her bone marrow, in addition to dysplastic features such as bi-and multi-nucleation of late normoblasts and internuclear bridging. She also showed 22% of blasts in her bone marrow cells thus classified in the category of RAEB-t after FAB classification. Also found were a chromosomal abnormality namely 46 XX t(14q21q), which was an unusual one. Her family brought her home and, 4 month later, she deceased. The second case, a 12 year-old boy, showed pancytopenia with Pseudo-Pelger-Huet anomaly in neutrophils but no blasts in the peripheral blood. Bone marrow of this case also showed the picture of dyserythropoiesis with binuclearity, internuclear bridging, nuclear fragments, multinuclearity of late normoblasts, in addition to megaloblastoid change in the normoblasts and increased mitosis in granulocytic series. No chromosomal abnormality was found. Classified as RA after FAB classification of MDS. Showed good response to conservative management for anemia for more than one year.

In-hospital mortality in patients with STEMI admitted during off hours

( Sung-Soo Kim ),( Myung Ho Jeong ),( Jeong Gwan Cho ),( Young Keun Ahn ),( Jong Hyun Kim ),( Shung Chull Chae ),( Young Jo Kim ),( Seung Ho Hur ),( In Whan Seong ),( Taek Jong Hong ),( Dong-hoon Choi 대한내과학회 2013 대한내과학회 추계학술대회 Vol.2013 No.1

Conflicting results exist on the outcome of off hours PCI in ST elevation myocardial infarction (STEMI). However, there were only a few studies that have focused on the clinical characteristics and outcomes of off hours PCI in STEMI. So, we studied the clinical characteristics and hospital mortality in STEMI patients treated with primary PCI during regular hours (weekdays 9:00 AM to 6:00 PM) versus off hours Weekdays 6:01 PM to 8:59 AM, weekends, and holidays) in Korea Acute Myocardial Infarction Registry. We analyzed in hospital and one year mortality among 5,665 consecutive ST segment elevation myocardial infarction patients treated with primary PCI between November 2005 to January 2008. Total 2,848 (50.2%) patients were treated during off hours. Baseline finding were similar, although regular hours patients were older. Median symptom to balloon time (304 min, IQR 175 to 750 vs. 270 min, IQR 145 to 551, p=0.001) were longer for regular hours primary PCI. Median door to balloon time (71min, IQR: 48 to 132 vs. 59 min, IQR 39 to 110 min, p=0.001) were longer for off hours pPCI. Also, Cardiac enzyme such as Max CK-MB (212.1±299.3 vs 194.7±303.4, p=0.031) and max TnI (72.6±239.5 vs. 58.9±94.4, p=0.013) were increased in off hours pPCI. However, unadjusted in hospital (6.0% off hours vs. 6.0% regular hours, p=0.946) and one year cardiac mortality (11.3% off hours vs. 11.7% regular hours, p=0.661) were comparable. In multivariate analysis, off hours primary PCI did not predict an adverse outcome. In conclusion, when primary PCI was performed within an appropriate reperfusion strategy, the clinical effectiveness of either off hours or regular hours pPCI is comparable.

Temporal trends and in-hospital outcomes of primary percutaneous coronary intervention in nonagenarians with ST-segment elevation myocardial infarction

( Joon Young Kim ),( Myung Ho Jeong ),( Yong Woo Choi ),( Yong Keun Ahn ),( Shung Chull Chae ),( Seung Ho Hur ),( Taek Jong Hong ),( Young Jo Kim ),( In Whan Seong ),( In Ho Chae ),( Myeong Chan Cho ) 대한내과학회 2015 The Korean Journal of Internal Medicine Vol.30 No.6

Background/Aims: Data regarding the outcomes of primary percutaneous coronary intervention (PCI) for ST-segment elevation myocardial infarction (STEMI) in nonagenarians are very limited. The aim of the present study was to evaluate the temporal trends and in-hospital outcomes of primary PCI in nonagenarian STEMI patients. Methods: We retrospectively reviewed data from the Korea Acute Myocardial Infarction Registry (KAMIR) from November 2005 to January 2008, and from the Korea Working Group on Myocardial Infarction (KorMI) from February 2008 to May 2010. Results: During this period, the proportion of nonagenarians among STEMI patients more than doubled (0.59% in KAMIR vs. 1.35% in KorMI), and the rate of use of primary PCI also increased (from 62.5% in KAMIR to 81.0% in KorMI). We identified 84 eligible study patients for which the overall in-hospital mortality rate was 21.4% (25.0% in KAMIR vs. 20.3% in KorMI, p = 0.919). Multivariate analysis identified two independent predictors of in-hospital mortality, namely a final Thrombolysis in Myocardial Infarction (TIMI) flow < 3 (odds ratio [OR], 13.7; 95% confidence interval [CI], 3.2 to 59.0; p < 0.001) and cardiogenic shock during hospitalization (OR, 6.7; 95% CI, 1.5 to 30.3; p = 0.013). Conclusions: The number of nonagenarian STEMI patients who have undergone primary PCI has increased. Although a final TIMI flow < 3 and cardiogenic shock are independent predictors of in-hospital mortality, primary PCI can be performed with a high success rate and an acceptable in-hospital mortality rate.

제주도 고등학교 학생들의 비만 정도, 심혈관 위험인자, 소아시 비만과의 관련성

홍성철,황승욱,현인숙 韓國保健敎育.健康增進學會 2003 보건교육건강증진학회지 Vol.20 No.2

Background: Recently, adolescent obesity was increased and tended to become obese adults, facing increased risk for diabetes, cardiovascular disease. Even before reaching adulthood, adolescent obesity already was experiencing hypertension, dyslipidemia, and diabetes mellitus. The purpose of this study was to find prevalence of obesity of high school students in Jeju island and association between obesity and cardiovascular risk factors and association between childhood obesity and adolescent obesity. Method: The study subjects were 6,064 students of twenty-two high school in Jeju island and health survey data was gathered from high school health examination data and hospital health examination data from Aug. 1999 to Dec. 1999. Adolescent obesity was evaluated by ideal body weight method in 1998 and body mass index. Cardiovascular risk factors were checked total cholesterol, systolic and diastolic blood pressure, fasting blood sugar. In 1,534 students, obesity at primary school is checked by primary and middle school health record. Results: Prevalence of obesity in Jeju island high school students is 12.6% in male students, 13.3% in female students by ideal body weight method and 8.7% in male students, 8.5% in female students by body mass index. Prevalence of obesity of male students in city area and district area of Jeju island were each 12.9%, 10.3% and that of male students in city area was significantly higher. Prevalence of obesity of female students in city area and district area of Jeju island were each 12.9%, 14.2% and not significantly different. Prevalence of obesity of male students in general and commercial school group were each 13.9%, 10.3% and that of general school group was significantly higher. In female students, Prevalence of obesity general and commercial school group was not significantly different. Total cholesterol, systolic and diastolic blood pressure were significantly associated with degree of obesity in male and female students(P<05) and fasting blood sugar was only significantly associated with degree of obesity in male students(P<05). In 1,514 students checked by health record, Odds ratio of obesity at primary and middle school that at high school were 12.96~22.77 and was significantly associated. Conclusion: Prevalence of obesity of high school students in Jeju island is very high and adolescent obsity was significantly associated with cardiovascular risk factors. Mostly childhood obesity tended to become adolescent obsity and program of control of obesity at childhood and adolescence is required.

제주시 일부 초, 중, 고등학생과 대학생들의 생체전기 저항법에 의한 체성분 조사

홍성철,이상이,현인숙 대한보건협회 2002 대한보건연구 Vol.28 No.2

Purpose : obesity is a frequent nutritional disorder in children and adolescent and its prevalence is increasing in Korea. The aim of this study was to measure body composition using bioelectical impedence in primaty, middle, high and college students in Cheju. Methods : We measured height, weight, and body composition(Fat %, WHR, Fat feee mass) by InBody 3.0 in 1,431 students aged 10-25 years. We also examined degree of obesity comparing standard weight for height, body mass index, and precent of body fat. Results : Fat free mass increased by aging in both sex, higher in male. Fat mass was highest in Primary school in male(12.09±4.86kg), college in female(16.03±4.22kg). Percent of body fat was highest in Primary student(21.64±8.02%), decreased middle(19.96±7.26%), high school(15.07±5.13%), and increased in college student(17.33±5.21%) in male, however, lowest in primary student(22.78±6.10%), increased by aging (middle;25.38±6.19%, high;27.61±5.27%, college;28.27±4.49%) in female. Percent of body fat was significantly higher in female except in primary school age. The prevalence of obesity based on fat % was highest in primary school student(primary, middle, high, college student; 33.3%, 23.3%, 7.6%, 8.2%) in male(≥25%), and highest in college student(11.1%, 26.6%, 32.3%, 38.6%) in female(≥30%), whereness, based on a standard weight for height(≥20%), 13.16%, 10.84%, 6.90%, 10.10% in male, 4.44%, 7.21%, 12.11%, 9.59% in female. Conclusion : It was necessary to evaluate obesity degree by body fat percentage, especially in adolescent and young female

적외선 센서 냉각용 맥동관 극저온 냉동기의 성능특성에 관한 실험적 연구

박성제(Seong-Je Park),홍용주(Yong-Ju Hong),고준석(Jun-Seok Ko),김효봉(Hyo-Bong Kim),염한길(Han-Kil Yeom),인세환(Se-Hwan In) 대한설비공학회 2016 대한설비공학회 학술발표대회논문집 Vol.2016 No.11

A pulse tube refrigerator is a small scale cryocooler which was invented by Gifford and Longsworth in 1963. It has the advantages of high reliability and low vibration because it has no moving part in the cold part. In recent years, the pulse tube cryocooler have cooling capacities and thermal efficiencies comparable with the Stirling cryocooler. These advantages promote the widespread use of the pulse tube cryocooler in many fields, specially in space-borne application that require a long life and stable temperature. An in-line pulse tube cryocooler has been designed and tested in our laboratory. The main part of the cryocooler, consisting of an in-line assembly of aftercooler, regenerator, cold heat exchanger, inertance tube, buffer and hot heat exchanger. Generally, a long tube with small diameter is used as an inertance tube and it imposes resistance and impedance to gas flow simultaneously. Phase between pressure and flow at cold-end is adjusted by length of the inertance tube with fixed diameter. This paper presents the results of a series of performance tests for the in-line type pulse tube cryocooler. Experimental investigations have been conducted to evaluate their performance characteristics with respect to several parameters such as the phase shifter, operating pressure and operating frequency of the linear compressor.

백서 뇌하수체 성장호르몬 종양세포의 Chicken Lysozyme 유전자 갑상선호르몬 반응요소에서 갑상선호르몬 수용체 역동학에 미치는 T₃효과 분석

이성진,박철영,정인경,홍은경,최철수,김현규,김두만,유재명,임성희,최문기,유형준,박성우,Larsen, P. Reed 대한내분비학회 2003 Endocrinology and metabolism Vol.18 No.4

연구배경: 유전자 전사과정은 증강부위 (enhancer) 또는 억제부위(silencer)의 복합작용을 통하여 조절되며 chicken Iysozyme 유전자의 억제부위는 두 개의독립적인 전사인자 결합부위 (Fl과 F2)를 가지는데 Fl부위는 75~93 kD 크기의 NePl 단백질이 결합하는 위치인 반면 역위회문구조(inverted palindrome, InvPal)의 F2 부위는 갑상선호르몬 수용체가 결합하는 갑상선호르몬 반응요소인 동시에 갑상선호르몬 수용체에 대해 높은 친화력을 가지고 있다. 실험적으로 Fl부위 또는 F2 부위 (이하 F2-TRE 부위)를 각각 다량체화(multimerization) 하였을 때 전사억제효과가 증가하였다는 연구 결과는 Fl 부위와 F2-TRE 부위가 서로 독립적으로 기능하는 구조임을 시사하고 있으며chicken Iysozyme 유전자의 억제부위가 완전한 전사억제효과를 가지기 위해서는 Fl 부위와 F2-TRE 부위가 모두 필요함이 보고 되어 있다. 현재 갑상선호르몬에 의한 chicken Iysozyme 유전자 조절기전을 규명하기 위해 많은 연구가 이루어지고 있으나 73 자극 전 ·후 갑상선호르몬 수용체의 역동학에 대해서는 아직까지 거의 보고된 바 없으며 이와 관련하여 저자들은 치근 사람의 간암세포주인 HepG2 세포에서 T₃ 자극전 ·후 갑상선호르몬 반응요소인 IRE2 부위에 대한 갑상선호르몬 수용체의 결합이 교대로 이루어지고 있음을 염색체 면역침전법 (chromatin immunoprecipitation, ChIP) 및 정량적 중합효소연쇄반응을 이용하여 확인한 바 있다. 이에 저자들은 본 연구에서 F2-lRE 부위를 포함하는 백서 뇌하수체 종양세포주인 GC8 세포를대상으로 염색체 면역침전법과 중합효소연쇄반응을 이용하여 갑상선호르몬 자극 전 후 시간적 순서에 따라 F2-TRE 부위에 결합하는 갑상선호르몬 수용체의 결합양상 변화를 분석함으로써 갑상선호르몬 수용체의 역동학적 모델을 제시하여 보고자 하였다. 방법: thymidine kinase(TK) promoter의 5' 부위에 chicken Iysozyme silencer의 고친화력 갑상선호르몬 반응요소(F2-TRE 부위)가 삽입된 플라스미드,mouse TRα gene이 삽입된 플라스미드, neomycinresistance gene이 삽입된 플라스미드를 백서 뇌하수체성장호르몬 종양세포인 GC 세포에 각각 주입하여 제작한 GC8 세포주를 사용하였다. 100 αM T₃를 투여하기 전과 투여한 후 12시간 뒤 TRαl, TRβl, TRβ2 항체를 이용하여 염색체 면역침전법과 고식적 중합효소연쇄반응 및 정량적 중합효소연쇄반응을 시행하였다. 각 갑상선호르몬 수용체 항체의 양을 1.5μL에서 4.5μL로 바꾸어 첨가한 후 동일한 방법으로 염색체 면역침전법을 반복하여 시행하였다. 100nM T₃를 투여하기전과 투여한 후 20분, 1시간, 2시간, 4시간, 6시간, 8시간, 12시간 뒤 TRαl, TRβl, TRβ2 항체를 이용하여 염색체 면역침전법을 시행하였다. 100nM T₃를 투여하기 전과 투여한 후 12시간 뒤 TRαl, TRβl, TRβ2 단백질의 발현량을 알아보고자 Western blot을 시행하였다. 결과: 100 nM T₃를 투여하기 전과 투여한 후 12시간 뒤 염색체 면역침전법과 F2-TRE 시발체를 이용한 고식적 중합효소연쇄반응을 시행하였을 때 T₃를 투여한 후 12시간 뒤 TRα1과 TRβ2의 결합은 증가한 반면 TRβl의 결합은 감소하였다. 정량적 중합효소연쇄반응으로 갑상선호르몬 수용체의 결합량을 측정하였을 때TRα1은 T₃ 투여 전 1.01에서 T₃ 투여 후 2.73으로 유의하게 증가하였으며 TBβl은 T₃ 투여 전 4.59에서 T₃투여 후 2.06으로 유의하게 감소하였고 TRβ2는 T₃ 투여 전 2.53에서 T₃ 투여 후 2.98로 증가하는 경향을보였다(TRα1, Δ=+170.3%, p<0.05; TRαl, Δ=-55.1%, p<0.05; TRβ2, Δ=+17.8%). 정량적 중합효소연쇄반응으로 측정한 갑상선호르몬 수용체의 전체 결합량은 T₃ 투여 전 8.13에서 T₃ 투여 후 7.77로 감소하였으나 통계적으로 유의하지 않았다(Δ=-4.4%).100nM T₃ 투여 전 ·후 시간별로 염색체 면역침전법과 정량적 중합효소연쇄반응을 시행하였을 때 TRα1 결합량은 T₃ 투여 후 20분과 6시간 뒤 각각 증가하였으며 TRβ2 결합량은 T₃ 투여 후 20분 뒤 최고치까지 증가하였다가 2시간 뒤부터 감소하였다. 그러나 TRαl 결합량은 T₃ 투여 후 1시간 뒤 최저치까지 감소되었다가 이후 지속적으로 유지되는 경향을 보였다. 100 nM T₃ 투여 전과 투여 후 2시간 뒤 갑상선호르몬 수용체의 결합량을 비교하였을 때 TRα1은 219.8% (1.01→3.23), TRβ2는 9.9% (2.53→2.78) 증가하였으나 TRβ1은 52.9% (4.59-)2.16) 감소하였으며 결합량 변화의 방향은 100 naM T₃ 투여 후 4시간 뒤와 6시간 뒤 갑상선호르몬 수용체 결합량 변화의 방향과 일치하였다(TRα1, 2.89→4.09, Δ =+41.5%; TRβl, 2.33→2.04, Δ=-12.4%; TRβ2, 2.57→2.59, Δ=10.8%). 갑상선호르몬 수용체 항체를 1.5 μL 또는 4.5 μL 투여한 후 F2-TRE부위에 대한 염색체 면역침전법 및 정량적 중합효소연쇄반응을 각각 시행하였을 때 첨가한 갑상선호르몬 수용체 항체의 양에 따른 갑상선호르몬 수용체결합량의 유의한 차이는 없었다. 100 nM T₃를 투여하기 전과 투여한 후 12시간 뒤 Western blot을 시행하였을 때 갑상선호르몬 수용체 발현량의 유의한 차이는 관찰되지 않았다. 결론: 본 연구에서 관찰된 T₃ 자극 전 · 후 chickenIysozyme 유전자의 F2-TBtE 부위에 대한 갑상선호르몬 수용체 이성체의 교대현상 및 시간적 순서에 따른 갑상선호르몬 수용체 결합양상의 변화가 나타내는 의미에 대하여 추시 연구가 필요함은 물론 추가적으로 다른 유전자 또는 다른 종류의 세포주를 대상으로 T₃자극에 따른 갑상선호르몬 수용체 결합양상의 변화와유전자 발현을 검토하여야 할 것이다. 한편 본 연구 결과만으로는 갑상선호르몬 수용체 역동학에 대한 많은 의문점을 풀 수 없음에도 불구하고 아직까지 국내외적으로 갑상선호르몬 수용체의 역동학에 대한 연구 결과가 거의 없는 현실을 고려하여 볼 때 본 연구는 제한적이나마 일정한 농도의 갑상선호르몬 자극 전ㆍ후chicken Iysozyme 유전자의 F2-TRE 부위에서 갑상선호르몬 수용체의 교대현상을 재확인하였다는 점과 갑상선호르몬 자극 전 · 후 시간적 순서에 따른 갑상선호르몬 수용체의 역동학적 모델을 처음으로 제시하였다는 점에서 의의가 있을 것으로 생각된다. Background: The regulation of gene transcription can be controlled by both positive (enhancer) and negative (silencer) regulatory sequences. Several enhancer and silencer elements have been described in the 5' region of the chicken lysozyme gene. The silencer located at -2.4 kb upstream of the chicken lysozyme gene is composed of two separate modules (Fl and F2) that can function as silencers by themselves, but also show synergistic repression after multimerization. The F1 module is bound by a protein termed NePl and F2 module, a F2 thyroid hormone response element (F2-TRE), and can be bound by the thyroid hormone receptor (TR). F2-TRE has an inverted palindromic structure, with high affinity to TR. Although many current reported results have tried to explain the regulatory mechanism of chicken lysozyme gene expression due to the thyroid hormone, there have been few studies that clarify the TR dynamics in the F2-TRE of the chicken lysozyme gene, either with or without exposure of the thyroid hormone. Here, the changes in the TR binding patterns in the F2-TRE of the chicken lysozyme gene are described, both before and after T₃ stimulation over time. Methods: Using the stably transfected rat pituitary somatotroph tumor cell line, GC8 cells, with the F2-TRE inserted 5' to the thymidine kinase (TIC) promoter, together with a mouse TRα - expressing plasmid, a chromatin immunoprecipitation (ChIP) technique was employed to reveal the TR-TRE interaction before and after T₃ stimulation. Following the cross-linking and sonication of the cells, the immunoprecipitation was performed overnight, at 4℃, with TRαl, TRβl and TRβ2 antibodies, respectively. The binding patterns and amounts of TRαl, TRβ1 and TRβ2 to the F2-TRE, before and after 12 hours of 100nM T₃ stimulation, were analyzed using conventional and quantitative real-time polymerase chain reactions (RQ-PCR). The ChIP technique was used to give a basal value for 20 minutes and 1, 2, 4, 6, 8 and 12 hours after the 100nM T₃ stimulation, and RQ-PCR was then performed. Western blot with TRαl, TRβl and TRβ2 antibodies were also performed. Results: After 12 hours of 100 nM T₃ stimulation of the GC8 cells, the TRα1 and TRβ2 binding to the F2-TRE increased, but the TRβ1 binding to the F2-TRE decreased, by conventional PCR. Although all the TR isoforms were bound to the F2-TRE by RQ-PCR, the TRαl binding to the F2-TRE, after 12 hours of l00nM T₃ stimulation, was significantly increased (1.01→2.73, Δ =+170.3%, p<0.05), but the change in the amount of TW2 binding was not significant (2.53→2.98, Δ=+17.8%). The TRβl binding was significantly decreased compared with that of the basal level (4.59→2.06, Δ=-55.1%, p<0.05). The total TR bindings to the F2-TRE had a tendency to decrease after 12 hours of 100 nM T₃ stimulation (8.13→7.77, Δ=-4.4%). The binding patterns and amounts of TRαl, Tβl and Tβ2, both before and after the 100 nM T₃ stimulation, were also identified over time. While the TRβl bindings to the F2-TRE after 1 hour of l00nM T₃ stimulation were acutely reduced, those of the TRαl at 20 minutes and 6 hours were increased. The TRβ2 bindings showed a maximal increase at 20 minutes. The directions of the TR binding patterns, between the before and after 2 hours of 100nM T₃ stimulation, were identical to those for between 4 and 6 hours of T₃ stimulation. There was no significant difference in the TR bindings to the F2-TRE in relation to the amounts (1.5 vs. 4.5μ I) of TR antibodies used during the ChIP assays. The Western blots showed no significant change of the levels of each TR isoform proteins, either before or after 12 hours of exposure to 100nM T₃. Conclusion: These results show the dynamic binding patterns of the TR isoforms to the F2-TRE of the chicken lysozyme gene, both before and after T₃ stimulation, over time. Further investigation, however, will be needed to clarify the mechanisms of our observations. The ChIP technique may then be used to reveal the dynamic models of the cofactors, as well as TR isoforms, in the TR-regulated transcription machinery (J Kor SOC Endocrinol 18:379-391, 2003).

세균독소로 자극시킨 사람 섬유아 세포에서의 Interleukin-6와 Interleukin-8의 생성

정인교,양동규,홍시영,이성근,김욱규,김광혁,김종렬 大韓顎顔面成形再建外科學會 1999 Maxillofacial Plastic Reconstructive Surgery Vol.21 No.4

Cytokines are hormone-like proteins which mediate and regulate inflammatory and immune responses. Interleukin-6 (IL-6) is involved in the final differentiation of B cells into antibody-producing cells. Interleukin-8 (IL-8) is a neutrophil chemotactic factor that plays an important role in the recruitment of neutrophil to inflammatory loci. Inflammatory mediators by cells in the gingiva have been implicated in the initiation and progression of periodontitis and oral infection. The purpose of this study was conducted to investigate the effect of lipopolysaccharide (LPS), staphylococcus enterotoxin B (SEB) on production ofIL-6 and IL-8 by human gingival and facial dermal fibroblasts. Primary cultured human gingival and facial dermal fibroblasts were incubated with LPS(0.01, 0.1, 1.0㎍/ml), SEB (0.01, 0.1, 1.0㎍/ml) or LPS (0.1㎍/ml) plus SEB (0.1㎍/ml). Culture supernatants were collected at 24, 48, and 72 hrs and assessed for IL-6 and IL-8 production by enzyme-linked immunosorbent assay. IL-6 production in gingival fibroblasts stimulated with LPS was higher than that with SEB. IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-6 production in facial demal fibroblasts was increased only by stimulation with a high concertration of LPS (1.0㎍/ml). Its production in facial dermal fibroblasts by exposure with SEB was decreased in comparison with control, nontreated cells. Therefore, gingival fibroblasts showed higher sensitivity than facial dermal fibroblasts in response to low concentration of LPS. Also, IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB.IL-6 production in facial dermal fibroblasts was increased only by stimulation with a high concentration of LPS(1.0㎍/ml). Its production in facial dermal fibroblasts by exposure with SEB was decreased in comparison with control, nontreated cells. Therefore, gingival fibroblasts showed higher sensitvity than facial dermal fibroblasts in response to low concentration of LPS. Also, IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in gingival fibroblasts was enhanced greatly only by stimulation of high concentration of LPS (1.0㎍/ml). That by exposure with SEB was increased only in 24 hrs cultivation. IL-8 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in facial dermal fibroblasts was decreased by LPS and increased only in 48 hrs cultivation by SEB. IL-8 production by double exposure with LPS plus SEB was enhanced only in 48 hrs cultivation in comparison with single exposure of LPS or SEB. therefore, IL-6 and IL-8 production were released at various quantities according to bacterial toxin applied and site of fibroblast harvested. These results suggest that gingival fibroblasts may be concerned with IL-6 and IL-8 related inflammatory response more than facial dermal fibroblasts.

한국 일 도시지역 청소년의 우울 증상 유병률과 관련요인에 대한 연구

조성진,전홍진,김무진,김장규,김선욱,류인균,조맹제 大韓神經精神醫學會 2001 신경정신의학 Vol.40 No.4

연구 목적 : 한국의 일 도시지역에 거주하는 청소년의 우울 증상의 유병률과 관련요인을 밝힘으로써 청소년 정신건강 증진을 위한 기초 자료를 수집하기 위하여 실시되었다. 방 법 : 연구 대상은 부천시에 거주하는 중학교나 고등학교에 재학중인 청소년 71,102명 가운데에서 무작위 집락 표본 추출법에 의해 선정된 2,203명이었으며, 이들에 대해서 CES-D(the Center for Epidemiologic Studies Depression Scale)와 사회인구학적 정보에 대한 문항이 들어 있는 설문을 실시하였다. 설문은 직접 학교를 방문하여 수행하였다. 선정된 2,203명의 청소년 가운데에 CES-D문항을 완성한 학생은 1,972명으로 반응률은 89.5%였다. 결 과 : 1) CES-D의 절단점을 16점으로 하여 계산한 '가능한' 우울 증상의 유병률은 중학생의 경우에 남학생 34.6%, 여학생 44.7%였으며, 고등학생의 경우에는 남학생 43.8%, 여학생 49.6%였다. 2) CES-D의 절단점을 25점으로 하여 계산한 '명확한' 우울증 증상의 유별률은 중학생의 경우에 남학생 18.4%, 여학생 20.7%였으며, 고등학생의 경우에는 남학생 21.4%, 여학생 23.9%였다. 3)종교, 인문계와 실업계의 여부에 따라서 우울 증상의 유병률에는 유의한 차이가 없었다. 4) 성,학년, 생활수준에 대한 자신의 평가, 가족의 구조, 성적에 대한 만족도의 다섯 가지 변수에 대해 CES-D점수가 16점 이상인 우울 증상의 유무에 따라서 로지스틱 회귀 분석을 시행하였을 때 성적에 대해 불만적 하는 군에서 매우 만족하는 군에 비해서 8.850배의 상대 위험도를 보였다. 생활 수준을 중하정도라고 대답한 군은 상이라고 대답한 군에 비해서 2.007배의 상대 위험도를 보였다. 여학생은 남학생에 비해서 1.307배의 상대 위험도를 보였다. 5) 우울 증상에 있어서 남학생과 여학생의 비율은 전체 군에서 1:1.38이며 중학생의 경우에는 1:1.88, 고등학생의 경우에는 1:1.13으로 나타났다. 고등학생에 있어 남학생의 우울 증상이 증가하는 것은 학업과 입시에 대한 부담과 관련된 것으로 생각되었다. 결 론 : 한국 일 도시거주 중고등학생에서 CES-D로 측정한 우울 증상은 남학생의 34.3%, 여학생의 47.5%에서 나타날 정도로 흔하다. 특히 학업 성적에 대해 불만족 하는 군에서 높은 우울 증상의 상대 위험률을 보이며, 고등학생에서 남학생의 우울증의 유병률이 높아지는 것은 이 지역 학생들이 학업에 대한 높은 부담을 가지고 있다는 것을 알 수 있었다. 따라서 이들에 대한 특별한 관심이 요구된다. Objective : We tried to see the prevalence and correlates of symptoms of depression in an adolescent population in Korea, and collected the basal data for mental health promotion of adolescents. Method : We sampled 2,203 adolescents among 71,102 adolescents living in Puchon City in Korea by the randomized clustered sampling method. The Center for Epidemiologic Studies Depression scale(CES-D) was used to measure depressive symptoms. The measurements were conducted through visiting schools. Of the 2,203 adolescents(aged 13 to 18) who were sampled, 1,972 persons completed CES-D and sociodemographic questions. The response rate was 89.5%. Result : 1) The prevalence rate of 'probable' depressive symptoms with cutoff point 16 was 34.3% in the boys and 47.5% in the girls. 2) The prevalence rate of 'definite' depression symptoms with cutoff point 25 was 17.4% in the boys and 20.6% in the girls. 3) We could not find any meaningful difference in the average of CES-D from the districts where they live, the course of their education, and their religions. 4) We used the logistic regression analysis to find the risk factor for adolescent depression in these samples. Among the variables degree, it was most important risk factor in adolescent depression to dissatisfy with their school degrees. Compared it with fully satisfied group, the odds ratio was 8.850. The group of mid to low socioeconomic status had the odds ratio 2.007 compared with high socioeconomic status. The girls had the odds ratio 1.307 compared with the boys. 5) The male versus female ratio was 1: 1.38 in the total students and 1 : 1.88 in the middle school students, and 1 : 1.13 in the high school students. The male versus female ratio was relatively low in the high school students. It was the result of the high prevalence of depressive symptoms in the high school boys, and it was due to the large burden to their school degrees. Conclusion : The depressive symptoms checked by CES-D were very common among adolescents in Korean urban area. The prevalence of depressive symptoms was 34.3% in the boys and 47.5% in the girls. The result that male versus female ratio was relatively low in the high school adolescents, and the group that were dissatisfied with the school degrees had high risk of depressive symptoms, revealed that the students had large burden to their school degrees and so they needed special concerns.