사면의 안정도 해석을 위한 입체투영법

이성대,이철우,이동우,이성규 公州大學校 基礎科學硏究所 1996 自然科學硏究 Vol.5 No.-

암반 사면의 안정성을 해석하는데 입체투영기법을 이용하였다. 안정도에 영향을 미치는 요인으로는 절리의 경사, 암석의 밀도, 내부마찰각등이 있으나, 가장 중요한 요인은 절리면의 경사이다. 이 연구에서는 암석의 점착력은 무시하고 단지 내부마찰각과 절리의 주향 및 경사만을 이용하였다. 그 결과 입체투영도에서 나타나는 암반사면의 안정도는 이동력과 저항력의 상대적인 크기에 의하여 좌우되는 것으로 나타났다. 동시에, 지반진동에 의한 영향도 입체 투영기법으로 찾아낼수 있다는 것이 밝혀졌다. Stereographic projection technique was used to analyze rock slope stability. Strike and dip of joints, rock strength, and internal frictional angle are major factor on slope stability. Among them, the most important factor is strike and dip of joints. In stereographgic projection technique, only internal frictional angle, and strike and dip of joints were used to interprete slope stability. Cohesion of rock was not considered. It is suggested that the rock slope stability on stereographic projection depends upon only the differential stress between driving and resisting force. At the same time, Stereographic projection technique on seismic loading was proposed in this study.

납 폭로 근로자에서 δ-aminolevulinic acid dehydratase 유전형질이 zinc protoporphyrin 및 δ-aminolevulinic acid에 미치는 영향

이병국,황규윤,장봉기,김용배,리갑수,안현철,김화성,이성수,안규동,박종범 순천향대학교 산업의학연구소 1999 순천향산업의학 Vol.5 No.1

The inhibition of δ-aminolevulinic acid dehydratase (ALAD) in the heme synthetic pathway results in increased protophoryphyrin (ZPP) and δ-aminolevulinic acid (ALA) and is responsible for some of toxicological effects of lead. This enzyme is coded by the ALAD gene containing 2 co-dominant alleles. The polymorphisms of ALAD gene may be related to differential effects of lead on ZPP and ALA, ALAD genotype was measured in 975 Korean male lead workers, of whom 897 were homozygous for ALAD1 (ALAD 1-1 genotype) and 96 were heterozygous for ALAD2 (ALAD 1-2 genotype). Blood lead in subjects with ALAD1 was significantly higher than those with ALAD2 (p = 0.01). No difference between ALAD genotypes was found for age, exposure duration, ZPP, ALA, hemoglobin, hematocrit, body mass index, tobacco and alcohol use. After adjustment for possible confounders, ALA and ZPP became significantly elevated in ALAD1 subjects (p = 0.004 and 0.055, respectively). This result suggests that ALAD1 subjects may be more susceptible to the hematotoxicologic effects of lead than ALAD2 subjects.

업종에 따른 연 취급 사업장의 기중 연 농도 및 연 노출 수준 평가

이병국,김용배,리갑수,안현철,김화성,이용진,황규윤,장봉기,이성수,안규동 순천향대학교 산업의학연구소 1999 순천향산업의학 Vol.5 No.1

In order to obtain an useful information for health management and biological monitoring of lead exposed workers, Authors tried to investigate air lead level and the lead exposed level of workers in lead industry according to occupational category. The subjects in our study were 2074 workers in 7 lead-using industries, and study subjects were divided into 4 occupational categories such as storage battery industry (type 1), primary smelting industry (type 2), secondary smelting industry (type 3) and litharge making industry (type 4). Blood zincprotoporphyrin concentration (ZPP), blood lead concentration (PbB) and urinary δ -aminolevulinic acid (ALAU), hemoglobin (Hb), hematocrit (Hct) were selected as the indices of lead exposure. Personal variables such as age, work duration were also collected. The results were as follows. 1. The geometric mean air lead in 9 lead-using industry was 0.1133±4.3120 ㎎/㎥, and that in type 1, 2, 3 and 4 was 0.1038±3.4952 ㎎/㎥, 0.0429±3.4329 ㎎/㎥, 0.1877±2.5123 ㎎/㎥ and 0.9961±5.2910 ㎎/㎥, respectively. 2. The mean ZPP in 9 lead-using industry was 53.1±28.0 ㎍/㎗, and that in type 1, 2, 3 and 4 was 52.0±24.8 ㎍/㎗, 48.0±16.4 ㎍/㎗, 109.8±85.6 ㎍/㎗ and 74.3±37.8 ㎍/㎗, respectively. There was significant difference in ZPP according to occupational category (P<0.01). The percents of lead workers whose ZPP were above 100 ㎍/㎗ in type 1, 2, 3 and 4 were 4.0%, 1.7%, 34.3% and 21.6%, respectively. 3. The mean PbB in lead-using industry was 26.0±11.2 ㎍/㎗, and that in type 1, 2, 3 and 4 was 25.3±11.1 ㎍/㎗, 26.7±8.8 ㎍/㎗, 50.3±15.2 ㎍/㎗ and 36.4±11.0 ㎍/㎗, respectively. There was significant difference in PbB according to occupational category (P<0.01). The percents of lead workers whose PbB were above 40 ㎍/㎗ in type 1, 2, 3 and 4 were 10.4%, 7.7% 71.9% and 43.2%, respectively. 4. While the correlation of Hb corrected PbB with ZPP was higher than non corrected PbB, the correlation of log-transformed ZPP with PbB was higher than non corrected ZPP. 5. Simple linear regressions of PbB and corrected PbB as independent variable with ZPP, log-transformed ZPP and ALAU as dependent variable were statistically significant (P0.01). Coefficient of determination of corrected PbB with other variables was higher than non corrected PbB. As the result of this study, it was suggested that reconsideration of environmental and biological monitoring program was highly recommended for secondary smelting and litharge making industry.

Hepatic Cirrhosis Occurring in a Young Woodchuck(Marmota monax) Due to Vertical Transmission of Woodchuck Hepatitis Virus(WHV)

Kyu-shik Jeong,정원일,Jae-yong Chung,Mi-young An,Chae-yong Jung,Gyoung-jae Lee,Jong-soo Kang,Byeong-cheol Kang,Young-heun Jee,Bruce H Williams,Young-oh Kwon,Da-hee Jeong 대한수의학회 2003 Journal of Veterinary Science Vol.4 No.2

Cirrhosis Occurring in a Young Woodchuck (Marmota monax) Due to Vertical Transmission of Woodchuck Hepatitis Virus (WHV)Da-hee Jeong, Won-il Jeong, Jae-yong Chung, Mi-young An, Chae-yong Jung, Gyoung-jae Lee1, Jong-soo Kang1, Byeong-cheol Kang2, Young-heun Jee3, Bruce H Williams4, Young-oh Kwon5 and Kyu-shik Jeong*College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Korea1Shinwon Scientific Co., LTD, Research Institute, Suwon, Korea2Clinical Research Institute, Seoul National University, Seoul 110-744, Korea3College of Veterinary Medicine, Jeju National University, Jeju 690-756, Korea

납노출지표와 적혈구내 protein kinase C 활성도의 연관성

황규윤,황보영,안현철,김용배,리갑수,이성수,안규동,이병국 大韓産業醫學會 2001 대한직업환경의학회지 Vol.13 No.4

목적 : Protein kinase C는 칼슘-인지질 의존형인산화 효소로 'H VJ'tfO에서 납에 의하여 활성화되지만, 납에 노출된 인체내에서 그 영향을 평가한 경우는 얼다. 본 연구의 목적은 납에 직업적으로 노출되는 근로자를 대상으로 납 노출이 적혈구막의 단백질내에서 PKC 활성에 의한 인산화 수준에 영향을 주는지 알아보고자 하였다. 방법 : 1998년부터 국내 납 근로자를 대상으로 납에 의한 건강 영향조사가 4년간의 코호트 연구로 실행하였다. 1차년도에 630명의 납 근로자와 135명의대조군이 조사되었고 이들중 본 연구에서는 직업적노출이 되는 사업장에 근무하는 212명의 근로자를대상하였다. 156명의 남자와 56명의 여자 근로자를 대상으로 인구학적, 과거병력, 직업력 등을 구조화된 설문과 면접으로 조사되었고 납 노출 평가는 혈중 납농도 및 ZPP, 경골중 납농도를 측정하였다. PKC의 활성도는 적혈구막 단백질내 PKC 의존형인산화 수준으로 평가하였다. 적혈구막 단백질인spectrin과 band 4.9의 후인산화수준을 측정하여각 납 노출지표(혈중 납, ZPP, 경골중 납. 노출기간)와의 관련성은 다중회귀분석을 이용하였다. 결과 : 조사대상자의 평균(SD) 연령은39.1(10.0)세, 근무기간은 8.1(6.5)년 이었으며, 경골중 납농도는 범위가 0.8에서 290.8 trg Pb/gbone mineral로 평근(SD) 34.4(35.2) rig Pb/gbone mineral이었다. 적혈구막 단백질의 후인산화수준은 개인간 변이가 매우 컸으며, spectrin은 평균(SD) 540 7(304.1), band 4.9 SfkDa는198.6(78.2), 48 kDa는 247.7(83.3) PSL이었다.경골중 납농도와 노출 기간은 이들 후인산화 수준과역상관성을 보였으나(p(0.05), 혈중 납 농도와 ZPP는 상관성이 없었다(p)0.05). 가능성 있는 혼란변수를 통제한 상태에서도 경골중 납 농도와 노출기간은 이들 후인산화수준과 유의한 회귀계수를 나타내었다. 결론 : 만성적 납 노출에 의하여 적혈구내 PKC활성도는 영향을 받아 증가되어있는 것으로 평가되어 적혈구막 단백질의 인산화수준은 납의 노출지표로 이용될 수 있을 것이다. 납의 신경독성은 부분적으로 PKC의 활성도왁 관련되어 있을 기전을 배제하기 어렵기 때문에 PfC 활성도와 신경행동학적 기능과의 관련성 평가가 진행되어야 할 것이다. Objectives : Protein kinase C(PKC) , a calcium and phospholipid dependent enzyme, is activated by lead in vitro at picomolar concentrations. However, the effect of lead on PKC has never been studied in a human population. The purpose of the study was to evaluate whether lead exposure was associated with PKC mediated-phosphorylation in erythrocytes among lead workers. Methods : Two hundred and twelve lead workers were studied. To determine the levels of phosphorylation ir vivo, an in vivo back phosphorylation technique was used by adding PKC and γ-32P to preparations of erythrocyte membranes. We measured back phosphorylations of erythrocyte membrane proteins, spectrin, and 52 kDa and 48 kDa, as an indirect measure of PKC activation if vivo. Results : The mean(SD) age and exposure duration was 39.1(10.0) years and 8.1(6.5) years, respectively. Tibial lead ranged from 0.8 to 290.8 μg Pb/g bone mineral with a mean (SD) of 34.4(35.2) μg Pb/g bone mineral. The means(SD) of back phosphorylation levels of the three proteins were 540.7(304.1), 198.6(78.2), and 247.7(83.3) photostimulated luminescence units (PSL), respectively, by phosphoimager. After adjustment for potential confounding factors, tibial lead and exposure duration were significantly and inversely associated with back phosphorylation levels. One unit of increase in tibial lead (1 μg Pb/g bone mineral) is associated with a decline in spectrin, band 4.9 52 kDa, and band 4.9 48 kDa back phosphorylation levels by 1.4(p〈0.05), 0.34(p〈0.05), and 0.47(p〈0.01), respectively However, there were no associations between the back phosphorylation levels and either blood lead or ZPP levels. Conclusions : These findings suggest that the PKC activity in erythrocytes is increased by chronic lead exposure and that erythrocyte membrane protein phosphorylation may be a biomarker of lead exposure.

한국재래흑염소에서 발정 및 과배란 유도와 외래유전자 주입에 적합한 1세포기 수정란의 채취

신상태,이두환,김명철,이운규,이철상,한용만,이경광 충남대학교 수의과대학 동물의과학연구소 1998 動物醫科學硏究誌 Vol.6 No.-

Three different treatments for induction of estrus in Korean native black goats were compared: follicle stimulating hormone(FSH, FSH-p^TM), FSH combined with MAP(intravaginal impregnated sponges, Veramix^ⓡ, containing 60 ㎎ medroxy progesterone acetate for 14 days), and FSH combined with progesterone(Ovaron^ⓡ, 10 ㎎ IM for 10 days) and PGF_2α(Lutalyse^ⓡ, 3 ㎎ IM at first FSH injection). FSH for inducing estrus and superovulation was given a total 20 ㎎ intramuscularilly in decreasing dosage injections twice daily over 4 days. The MAPs were withdrawn at the 3rd day of FSH injection. Estrus observations were conducted every 6 hours from last FSH injection for 24 hours by placing the does with fertile male goats. Estrus and superovulation were more successfully induced with treatment of MAP + FSH than other treatments(FSH only, or progesterone + PGF_2α + FSH) (estrus induction; 100 vs 42.8 and 71.4%, ovulation points; 11.4 vs 5.4 and 4.4, respectively). The effect of gonadotropin releasing hormone(GnRH) on the ovulation rate was also examined. However, no difference was observed for inducing ovulation with treatment or dosage(100 ㎍ 200 ㎍) of GnRH. Low midline laparotomies were performed, and then ovarian responses (ovulations and follicular development) were examined by exteriorization of the reproductive tracts. Ova were recovered from oviducts by retrograde flushing 60-146 hours after MAP removal, and were classified the developmental stages. Overall 66.1% (236/357) of recovery rate was obtained from 30 superovulated does. The optimal recovery time of microinjectable 1-cell zygotes was approximately 72-76 hours after MAP removal.

연취급 근로자들의 사회 심리적 스트레스와 연노출 지표들간의 관련성

김용배,안현철,황보영,리갑수,황규윤,이성수,안규동,이병국 大韓産業醫學會 2000 대한직업환경의학회지 Vol.12 No.1

목 적 : 연취급 근로자를 대상으로 사회 심리적 스트레스 수준을 알아보고 이를 기초자료로 활용하며 사회 심리적 스트레스와 연노출 지표들간의 관련성을 알아보고자 시도하였다. 방 법 ' 1차 연제련업종의 근로자 234명을 대상으로 일반적 특성, 사회 심리적 스트레스, 직업적 특성과 사회적 지지를 문진과 설문지를 통해 확인했으며 연노출 수준을 알기 위해 혈중 ZPP 농도, 혈중연농도, 요중 δ-ALA 배설량, 혈색소, 혈구용적치등을 측정했다. 결 과 : 단일변량 분석결과 혈중 연농도 수준과 직업적 특성에 따른 사회 심리적 스트레스는 유의한 차이가 있었으며 혈중 연농도가 증가함에 따라 스트레스 수준도 증가되는 경향을 나타냈다. 사회 심리적 스트레스와 연노출 지표 및 다른 변수들간의 관련성을 파악하기 위하여 상관분석을 실시한 결과 혈중 연농도만이 사회 심리적 스트레스와 양의 상관관계를 나타냈으며 유의했다. 결 론 : 고농도 연노출의 가능성이 있는 직종인 경우 기존의 연관련 자각증상에 대한 설문조사와 함께 사회 심리적 스트레스 측정도 병행하여 실시하는 것이 연에 의한 건강장해를 조기에 발견하고 예방하는데 필요하다. Objectives : This study was carried out to assess psychosocial distress of lead workers and to examine the relationship between lead exposure indices and psychosocial distress. Methods The study design was cross-sectional and research subjects were 234 lead workers in primary smelting industry. General characteristics, job characteristics and social support were obtained by self-administered questionnaire and history taking. Psychosocial distress was measured using PWI(Psychosocial Well-being Index), a 45-item self-administered instrument. Blood lead(PbB), zinc protoporphyrin(ZPP), δ-aminolevulinic acid in urine (ALAU) were selected as indicators of lead exposure. Results : There was significant difference in PWI according to lead exposure level and job characteristics. The results of correlation analysis showed that PWI was significantly correlated with PbB (r=0.203, p=0.002). Conclusions : Our results supported the association between lead exposure and psy-chosocial distress. The inclusion of psychosoclal distress measurement as a special health examination for lead workers was highly recommended for the effective health manage-ment.

직업적 연폭로에서 혈중 연량 수준에 따른 선별검사로서의 혈중 ZPP 검사의 타당도

한구석,안현철,김용배,리갑수,김화성,황규윤,장봉기,이성수,안규동,이병국 순천향대학교 산업의학연구소 1999 순천향산업의학 Vol.5 No.1

In order to evaluate the validity of current screening criteria of lead exposure and to provide effective information for the better health management of lead workers, author investigated the interrelationship between blood lead and blood zinc protoporphyrin(ZPP) and evaluate the validity of blood zinc protoporphyrin as the screening test and blood lead as gold standard of diagnostic test. The total of 2245 lead workers (male workers: 1982, female workers : 263) from 12 lead using industries were studied. Blood lead, ZPP, hemoglobin and hematocrit were selected as lead exposure variables. Personal information on sex, age and work duration were also checked. Blood lead was measured with flameless atomic absorption spectrometry (model: Hitachi 8100) using standard addition method. Blood ZPP was measured with portable hematoflurometer (model: Aviv 206). The results obtained were as follows: 1. The mean blood lead of all lead workers were 0.7±14.3㎍/㎗(Male: 31.0±14.4㎍/㎗, Female: 28.2±12.9㎍/㎗), whereas the mean blood ZPP of those were 68.1±47.5㎍/㎗(Male: 63.9±43.7㎍/㎗, Female: 100.1±61.0㎍/㎗). The mean value of hemoglobin and hematocrit of all lead workers were 14.5±1.5g/㎗(Male: 14.7±1.3g/㎗, Female: 12.3±1.2g/㎗) and 45.4±3.6(Male: 46.2±2.8, Female: 39.5±3.1) respectively. 2. Validity of test using blood zinc protoporphyrin as screening test for blood lead levels of 30㎍/㎗ as gold standard of diagnostic test revealed that the highest validity value for male and female lead workers were 50 and 90 ㎍/㎗ of ZPP respectively, showing 50 and 60 ㎍/㎗ of ZPP for all lead workers. While those test for blood lead levels of 40㎍/㎗ as gold standard of diagnostic test revealed that the highest validity value for male and female lead workers were 60 and 90 ㎍/㎗ of ZPP respectively, showing 60 ㎍/㎗ of ZPP for all lead workers, those test for blood lead levels of 50 and 60㎍/㎗ as gold standard of diagnostic test revealed that the highest validity value for male and female lead workers were 60 or 70 and 100 ㎍/㎗ ZPP respectively, showing 60and 70 or 80 ㎍/㎗ of ZPP for all lead workers. 3. Predictive value of test using blood zinc protoporphyrin as the screening test for different blood lead levels of 30, 40, 50 and 60㎍/㎗ as gold standard of diagnostic test revealed that positive predictive value of screening test were decreased by the increase of blood lead level regardless of csreening criteria value ZPP and negative predictive value of screening test were increased accordingly regardless of screening criteria value of ZPP. On the other hand, within same blood lead level as a gold standard positive predictive values were inceased by the increase of screening criteria value of ZPP, whereas the negative predictive value were decreased accordingly by the the increase of screening criteria value of ZPP. 4. The correlation analysis of all lead workers among study variables revealed that CPbB and natural log transformation ZPP(LZPP) were better correlated with other variables and found to be better estimate than PbB and ZPP themselves. The age was correlated with all other variables, but the work duration was not. The correlation analysis of male and female workers among study variables revealed the same results with all lead workers. 5. The slope of simple linear regression of CPbB (independent variable) with ZPP in female workers(3.223)was steeper than in male workers(2.144). That of lead workers with work duration of less than 1 year was steeper than that of lead workers whose work duration was more than 1 year. But there was no slope difference between two groups divided by their work duration of 2 years less and more. The slope of simple linear regression of CPbB lead (independent variable)with ZPP in lead workers whose blood lead were higher than 30㎍/㎗and 40㎍/㎗ were steeper than those whose blood lead were less than 30㎍/㎗ and 40㎍/㎗. 6. The slope of simple linear regression of CPbB (independent variable) with ZPP in lead industries with low mean blood lead were less steeper than those with high mean blood lead. 7. The slope of simple linear regression of CPbB (independent variable) with LZPP in female workers(0.0308was steeper than in male workers(0.0253). That of lead workers with work duration of less than 1 year was steeper than that of lead workers whose work duration was more than 1 year. But where was no slope difference between two groups divided by their work duration of 2 years less and more. While the slope of simple linear regression of CPbB lead (independent variable) with LZPP in lead workers whose blood lead were hogher than 30㎍/㎗ was not steeper than those whose blood lead were less than 30㎍/㎗, there was no slope difference between two groups whose blood lead were less than 40㎍/㎗ and more than 40㎍/㎗. 8. The slope of simple linear regression of CPbB (independent variable) with LZPP in lead industries with low mean blood lead were less steeper than those with high mean blood lead, but the difference was not significant then that of CPbB and ZPP. With above results, it is recommended that the screening criteria value of ZPP should be down to 60-70㎍/㎗ from 100㎍/㎗ for male lead workers and screening criteria of ZPP for female lead workers was all right with current value of 100㎍/㎗. So it is recommended to make different screening criteria of ZPP for male and female workers like the hemoglobin and hematocrit screening criteria. It is also preferable to use CPbB and LZPP rather than uncorrected PbB and ZPP in the evaluation of lead exposure for lead workers.

제1형 탈요오드효소 유전자 갑상선호르몬 반응요소에서 T₃자극에 따른 갑상선호르몬 수용체 역동학 모델

이성진,박철영,정인경,홍은경,최철수,김현규,김두만,유재명,임성희,최문기,유형준,박성우,Larsen, P. Reed 대한내분비학회 2003 Endocrinology and metabolism Vol.18 No.3

연구배경: 제1형 탈요오드효소의 발현에 관여하는 hdiol 유전자는 5 flanking region 내 서로 다른 특성을 가진 두 종류의 갑상선호르몬 반응요소, 즉 TREI과 TRE2를 가지고 있음이 알려져 있다. 사람의 간암세포주인 HepG2 세포에서 T₃를 투여하였을 때 hdiol유전자의 전사작용이 급격하게 증가하는데 hdiol mRNA가 충분히 발현하기 위해서는 두 종류의 갑상선호르몬 반응요소가 모두 필요함이 보고 되어 있으나 T₃ 투여시 갑상선호르몬 반응요소와 결합하는 갑상선호르몬 수용체의 역동학에 대해서는 아직까지 연구된바 없다. 한편 현재까지 보고 된 연구 결과들은 갑상선호르몬 자극이 없더라도 갑상선호르몬 수용체와 갑상선호르몬 반응요소가 서로 지속적으로 상호작용 한다는 전제 조건을 바탕으로 하고 있는데 아직까지 이러한 가정은 간접적으로만 증명되어 있는 상태이다. 이에 저자들은 본 연구에서 사람의 간암세포주인 HepG2세포를 대상으로 염색체 면역침전법과 중합효소연쇄반응을 이용하여 갑상선호르몬 자극 전·후 hdiol 유전자의 갑상선호르몬 반응요소에 결합하는 갑상선호르몬 수용체의 결합양상 변화를 분석함과 동시에 갑상선호르몬 자극 전에도 갑상선호르몬 수용체와 갑상선호르몬 반응요소가 서로 결합된 상태로 존재함을 직접적으로 확인하고자 하였다. 방법: 사람의 간암세포주인 HepG2 세포를 대상으로 100 nM T₃를 투여하기 전과 투여한 후 12시간 뒤 TRα1, TR 1, TR 2 항체와 IREI, TRE2에 상보적인 시발체를 이용하여 염색체 면역침전법과 고식적 중합효소연쇄반응 및 정량적 중합효소연쇄반응을 시행하였다. 100nM T₃를 투여하기 전과 투여한 후 12시간 뒤 hdiol mRNA 발현량의 변화를 정량적으로 측정하기 위하여 역전사 중합효소연쇄반응을 시행하였다. 100 nM T₃를 투여하기 전과 투여한 후 12시간 뒤 TR4'1, TR 1, TR 2 단백질의 발현량을 알아보고자Western blot을 시행하였다. 결과: 100 nM T₃를 투여하기 전과 투여한 후 12시간 뒤 염색체 면역침전법과 TREI 시발체를 이용한 고식적 중합효소연쇄반응을 시행하였을 때 T₃ 투여 전후 TREI 부위에는 TRgl이 결합하였으며 T₃를 투여한 후 TRal 결합이 감소하였다. 정량적 중합효소연쇄반응으로 TR4'1 결합량을 측정하였을 때 T₃ 투여 전3.74에서 T₃ 투여 후 1.97로 감소하여 통계적으로 유의한 차이를 나타내었다(Δ=-47.3%, p<0.05). T₃ 투여 전 ·후 TRβl과 TRβ2의 결합은 관찰되지 않았다. 100 nM T₃를 투여하기 전과 투여한 후 12시간 뒤 염색체 면역침전법과 TRE2 시발체를 이용한 고식적 중합효소연쇄반응을 시god하였을 때 T₃ 투여 전ㆍ후 TRE2 부위에는 TRα1, TR 1, TR 2가 모두 결합하였으며 T₃를 투여한 후 TRα1과 TR 1의 결합은 감소하였으나 TR 2의 결합은 증가하였다. 정량적 중합효소연쇄반응으로 갑상선호르몬 수용체의 결합량을 측정하였을 때 TRαl은 T₃ 투여 전 10.41에서 T₃ 투여 후 3.01, TRβl은 T₃ 투여 전 12.56에서 T₃ 투여 후 2.93으로 유의하게 감소하였으며 TRβ2는 T₃ 투여 전 9.17에서 T₃ 투여 후 9.84로 증가하는 경향을 보였다(TRα1, Δ=-71.1%, p<0.05; TR 1, Δ=-76.7%, p<0.05; TR2, Δ=+7.3%). 정량적 중합효소연쇄반응으로 측정한 갑상선호르몬 수용체의 전체 결합량은 T₃ 투여 전 32.14에서 T₃ 투여 후 15.78로 감소하여 통계적으로 유의한 차이를 나타내었다(Δ=-50.9%, p.0.05). 갑상선호르몬 수용체 항체를 1.5μL와 4.5μL 첨가한 후TREI과 TRE2에 대하여 염색체 면역침전법 및 정량적 중합효소연쇄반응을 각각 시깡하였을 때 첨가한 갑상선호르몬 수용체 항체의 양에 따른 갑상선호르몬 수용체 결합량의 차이는 없었다. 100 nM T₃를 투여하기전과 투여란 후 12시간 뒤 역전사 중합효소연쇄반응 및 hdiol cDNA 시발체를 이용한 정량적 중합효소연쇄반응을 시행하였을 때 T₃를 투여한 후 hdiol mRNA발현량은 2.03배 증가하였다(p<0.001). 100 nM T₃를 투여하기 전과 투여한 후 12시간 뒤 Western blot을 시행하였을 때 갑상선호르몬 수용체 발현량은 유의한 차이가 없었다. 결론: 현재까지 갑상선호르몬 수용체의 역동학에 대한 연구가거의 이루어지지 않았던 실정을고려하여볼 때 본 연구는 제한적이나마 일정한 농도의 갑상선호르몬 자극 전 · 후 갑상선호르몬 수용체 결합양상의 변화, 특히 T₃ 자극 전 hdiol 유전자의 TREI 부위에서 TRal이 억제자 (silencer)로서 작용할 가능성 및 T₃자극 전 · 투 TRE2 부위에서 갑상선호르몬 수용체 교대현상을 처음으로 제시하였다는 점에서 의의가 있으며 향후 다른 종류의 세포주 및 체내에서 갑상선호르몬 자극 전 후 갑상선호르몬 수용체 결합양상의 변화 및 유전자 발현에 미치는 영향을 연구할 필요가 있으리라 생각된다. 한편 염색체 면역침전법을 통해 HepG2 세포에서 T₃ 자극이 없더라도 갑상선호르몬 수용체와 갑상선호르몬 반응요소 사이에 지속적인 상호작용이 존재할 뿐 아니라 갑상선호르몬 반응요소에 대한 갑상선호르몬 수용체의 결합이 교대로 이루어지고 있음을 직접적으로 확인할 수 있었으며 향후 T₃ 자극 전 · 후 갑상선호르몬 수용체를 통한 유전자 전사조절기전에 관여하는 전사인자와 역동학적 기전을 규명함에 있어서 염색체 면역침전법과 정량적 중합효소연쇄반응이 매우 유용하게 이용될 수 있을 것이다. Background: Type 1 iodothyronine deiodinase (Dl), the product of the hdiol gene, is involved in thyroid hormone activation by the deiodination of thyroxine (T4) to form 3,5,3'-triiodothyronine (T3). Recent studies have identified two thyroid hormone response elements (TREs) in the 5 ' flanking region of the hdiol gene. TRE1, proximal to TRE in the hdiol gene, consists of a direct repeat of thyroid hormone receptor (TR) binding octamers with 10 bp separating the two TR binding sites. The upstream TRE, TRE2, is a classical direct repeat of retinoid X receptor (RXR)/TR binding half-sites with a 4-bp separation. There are few studies clarifying the TR dynamics in the TRE of a specific gene with or without the exposure of activated thyroid hormone. We evaluated TR binding patterns in the proximal and distal TREs of the hdiol gene before and after T₃ stimulation. Methods: We employed chromatin immunoprecipitation (ChIP) technique to investigate the TR- TRE interaction before and after T₃ stimulation in human hepatocellular carcinoma HepG2 cell line. Following cross-linking and sonication of the cells, immunoprecipitation was performed overnight at 4℃ with TRαl, TRβ1 and TRβ2 antibodies. We analyzed the binding patterns and amounts of TRαl, TRβl and TRβ2 to TREl and TRE2 before and after 12 hours stimulation with 100 nM T3 by using conventional and quantitative real-time polymerase chain reactions (RQ-PCR). Reverse transcriptional PCR (RT-PCR) and Western blot with TR 1, TR 1 and TR 2 antibodies were performed to measure the levels of hdiol mRNA and TR 1, TR 1 and TR 2 proteins before and after 12 hours exposure to l00nM T3. Results: In TRE1, TRαl binding was significantly decreased after 12 hours stimulation with l00nM T3 (3.74→97, Δ=-47.3%, p<0.05), but TRβ1 and TRβ2 bindings were not detected by conventional PCR and RQ-PCR. Although all TR isoforms were bound to TRE2, the binding patterns were quite different. While TRα1 and TRβ1 bindings to TRE2 after 12 hours stimulation with 100 nM T3 were significantly decreased (10.41→3.01, Δ=-71.1%, p<0.05; 12.56 →2.93, Δ =-76.7%, p<0.05, respectively), TRβ2 binding was increased but not significantly (9.17 →9.84, Δ =+7.3%). Total TR bindings in TRE2 were significantly decreased after 12 hours stimulation with 1OOnM T₃ (32.14 →15.78, Δ=-50.9%, p<0.05). The TR bindings to TREl and TRE2 were not significantly different by the amounts of TR antibodies used during ChIP assays. The levels of hdiol mRNA were significantly increased, 2.03 times, after 12 hours exposure to l00nM T3 (p<O.001). Western blot showed no significant change of the level of each TR isoform protein before and after 12 hours exposure to 100nM T3. Conclusion: Our results demonstrate the dynamics of TRal at proximal TRE (TRE1) and the switching phenomenon of TR isoforms at distal TRE (TRE2) of the hdiol gene after T3 stimulation. Further investigation, however, is needed to clarify the mechanisms of these observations (J Kor SOC Endocrinol 18:283-295, 2003).

8051을 이용한 새로운 검색 시스템의 구현

李俊榕,吳圭哲,金珍 弘益大學校 科學技術硏究所 2000 科學技術硏究論文集 Vol.11 No.-

Nowadays, there are large scale of creations, distributions and transformations of informations. Wide sharing and urgent changing of information is a primary characteristics of recent period. By the way, some kind of informations must be treated in different way. For example, geographic information for military operations must be distributed in great care with strict security and maybe few change will occur on such information. In this paper, we have studied and implemented new retrieval system that are especially appropriate when informations have following characteristics. 1. when consistency of information is important 2. when distribution of information must be done collectively 3. when change of information is not severe