설사유발 병원성 대장균에 관한 연구

박성철 김천대학교 1990 김천대학교 논문집 Vol.11 No.-

Enterotoxigenic Escherichia coli (ETEC) may induce Clinically significant intestinal fluid secretion (i.e. diarrhea) in humans and animals by production of enterotoxins. Several types of ETEC enterotoxins are recognized, each with a unique structure and mechanism of action. The native toxin is biologically stable after heat treatment at 30 and 100 C for 15 min, at pH range of 1-8 for 2hr, and after treatment with the proteolytic enzymes (pepsin, trypsin, pronase).

圓光醫大 附屬 第2病院 神經精神科 患者에 對한 統計的 考察(Ⅱ)

박민철,장광철,김상원,김한주,유미경,김훈,유용진,김현정,박남진,오상우,이귀행,노승호,백영석 圓光大學校 醫科大學 神經精神科學敎室 1992 圓光精神醫學 Vol.8 No.1

본 연구는 1986년 7월 11일부터 1991년 7월 10일까지 원광의대 부속 제2병원 신경정신과에서 의래 및 입원치료를 받았던 6,072명의 환자를 대상으로 병원 역학조사를 실시하고 그 결과를 1차조사 (박민철 등1986)와 비교했다. 1.전체 환자 중 남자가 54.4%로 여자보다 많았고, 이리와 이리 인접지역이 84.0%로 대부분의 지역을 나타났다. 의료보장상태는 보험환자가 57.1%, 의료보호환자21.8%, 일반환자 21.1%였으며 연령별로는 20대 24.6%, 30대 22.8%로 20대 30대가 절반정도를 차지했다. 2.외래환자에서는 신경중성장애가 46.0%로 가장 많고 정동장애, 경련성장애 순이며, 입원환자에서는 정신분열증이 30.5%로 가장 많고 기질성정신장애, 정동장애의 순이었다. 3.남자는 신경증성쟝애(23.4%), 정신분열증, 기질성정신장애 순이고 여자는 신경증성장애(39.9%), 정동장애, 정신분열증의 순이었다. 남자가 여자보다 많은 진단은 알코올정신장애(94.6%), 기질성정신장애, 소아정신장애 순이고, 여자가 남자보다 많은 진단은 정동장애(64.5%), 신경증성장애였다. 4.진단별 연령분포에서 신경증성장에는 30대(25.3%), 20대, 40대 순이고, 정신분열증은 20대(40.5%), 30대, 40대 순이며, 정동장애는 20대, 30대, 50대 순이나 비교적 비슷한 융이었다. 5.진단별 발병연령에서 신경증성장에는 20대, 30대가 절반이고,정신분열증은 20대(51.3%), 10대, 30대 순이며 정동장애는 20대(30.7%), 30대, 50대 순이었다. 진단별 평균 발병연령은 신경증성장애 32.7세, 정동장애 36.1세, 정신분열증 23.6세였다. 6.진단별 이병기간은 2년이상이 신경증성장에 28.3%, 정신분열증 68.5% 정동장애 43.9%인데 전체적으로 41.0%였다. 진단별 평균 이병기간은 정신분열증 73.2개월, 정동장애 45.5개월, 신경증성장애 29.5개월이었다. We examined epidemiologic study of 6,072 patients who visited Wonkwang Neuropsychiatric hospital since July 11 of 1986 until July 10 of 1991 and compared this results with previous study(Park et al 1986). The study revealed as follows:1. Of total patients males visited this hosptial more than females as 57.1%, Most of them(84.0%) resided Iri and neighbouring Iri. In the point of insurance, insured patients, medicaid patients and uninsured patients were 57.1%, 21.8% and 21.1% respectively. On the distribution of age, the twenties, the thirties were 24.6%, 22.8% respectively and twenties and thirties were about half of total patients. 2. Of outpatients neurotic disorder was the most as 46.0%, secondly mood disorder and thirdly convulsive disorder, but of inpatients schizophrenia was the most as 30.5%, secondly organic mental disorder and thirdly mood disorder. 3. Of males neurotic disorder was the most as 23.4%, secondly schizophrenia, and thirdly organic mental disorder, but of females neurotic disorder was the most as 39.9%, secondly mood disorder and thirdly schizophrenia. alcoholic mental disorder(94.6%), organic mental disorder, child mental disorder were mostly found in males,but mood disorder(64.5%) and neurotic disorder were mostly found in females. 4. On the distribution of age neurotic disorder was found the most in the thirties(25.3%), next in the twenties and in the forties, and schizophrenia was found the most in the twenties(40.5%), next in the thirties and in the forties, but mood disorder was found the most in the twenties, next in the thirties and in the forties but revealed similar rates relatively. 5. On the distribution of age of onset neurotic disorder developed nearly half during the twrnties and the thirties, schizophrenia developed mostly during the twenties (51.3%), secondly the teens and thirdly the thirties, but mood disorder developed firstly the twenties(30.7%), secondly the thirties and thirdly the fifties. on the average age of onset of illness neurotic disorder, mood disorder and schizophrenia were 32.7 years, 36.1 years and 23.6 years respectively. 6. On the duration of illness more than two years was 28.3% in neurotic disorder, 68.5% in schizophrenia and 3.9% in mood disorder and on the average duration of illness schizophrenia, mood disorder and neurotic disorder were 73.2 months, 45.5 months and 29.5 months respectively.

間歇的 運動에 의한 脫水 및 水分攝取의 生理的 效果

朴哲浩,朴吉俊,李相于,金永明,朴贊熙,玄松子,呂南會,朴相甲,金榮俊 東亞大學校附設스포츠科學硏究所 1989 스포츠科學硏究論文集 Vol.7 No.-

This study aims at revealing the physiological responses of dehydration (DH) and rehydration (RH) in intermittent exercises at the work intensity of 70% HR max. First, with general student public, three times of exercise (each for 20 minutes) at the given work intensity were taken, with a break for 25 minutes respectively. Their heart rate, rectal temperature and plasma electrolyte concentration both DH and RH were measured. Second, in two groups of athletes and non-athletes, 9 rounds of exercise (for 100 minutes) and 8 breaks (for 60 minutes) were taken, with RH forced 10 times to compensate for the subjects' water loss. Their change in sweat loss and plasma electrolyte concentration as well as the IVDP(Intensive Voluntary dehydration Phenomenon) were experimented. The summary of this research follows. 1) Mean heart rate and rectal temperature in intermittent exercises were higher at DH than at RH, growing up at the increasing level of DH. 2) RH saw no change in the plasma concentration of Na+, K+ and Cl-, while DH observed a significant increase. 3) HCO₃made a significant decrease at DH before growing up. 4) Mg++ had no change, while Ca++ increased significantly at RH and DH. 5) Lactate showed an increase with significance at RH and DH, especially greater at DH. 6) Osmotic pressure increased meaningfully at DH. 7) Anion gap increased significantly at DH and RH. 8) On the other hand, when the forced RH was held, the group of athletes had greater sweat loss per body surface area than that of non-athletes. 9) The athletes' plasma concentration of Na+ and Cl- revealed a clear decrease. 10) There was seen no significant change in the plasma concentration of K+, Mg++ and Ca++ in both groups. 11) The IVDP occurred at the RH of 2.3ℓfor athletes and more than 2.5ℓfor no-athletes.

地域經濟活性化를 위한 農村市場開發에 관한 硏究 : 忠南 禮山地域市場을 中心으로

尹畯相,李武鉉,申尹撤,洪性贊,朴奉圭 公州大學校 産業開發硏究所 1994 産業開發硏究 Vol.2 No.-

Since the first year economic development plan, imblanced development of national territory during the economic development process has aggravated remarkable disparity between urban and rural areas. For this reason, this study introduced the development of rural market as one of the development strategies of regional economic to correct the imblance and relieve regional disparity. The purpose of this study were to look into the present situation of traditional rural market and to analyze the problems for the rural development in view of the current road situations in the rural areas and status of marketing facilities in market center and to measure the degree of rural area people's satisfaction to the rural market and to suggest the development plan of activating for rural market center. To collect data, the Yesan Gun rural market were selected and a total of 591 residents and 178 merchants were sampled as final subjects of survey. Interview and/or questionnaire method was the main method of data collection. The major findings of this study were as follows: 1) The population of Yesan Gun has decreased 2.85 persons per year from in 1984, but the number of vehicles were increased 2.85 times than in 1989. And the parking capacity of Yesan Eup was only 129 vehicles which were below of the total vehicles of Yesan Gun area (14,352). The number of Yesan area markets were 8 markets which consisted of 1 permanent market and 7 periodic markets. But as the economy has been developed, the conditions of rural markets have been changing. That is, the role of periodic market tends to be weakened with the growth of large cities, reinforcement of permanent markets, development of traffic facilities, large production, preferring high quality goods. 3) Generally, the people of Yesan Gun were attached for purchasing the necessaries of life and comodities and farming materials to ① Yesan Eup ② Seoul ③ Chonan Shi ④ Hongsung gun ⑤ Daejeon Shi. 4) The degree of community people's satisfaction to the town market presented 26.6% which meaned higher than dissatisfaction(9.5%). And the degree of people's inconvenience of accessibility to market center by transferring of bus terminal presented 64.2% which meaned higher than average. 5) The degree of people's dissatisfaction of traffic system in Yesan gun area presented followings: the road networks(61.5%), parking facility(68%), bus route(54%). 6) By the survey of merchants of Yesan gun area, on the average, it showed followings: the size of store was 6-10 pyeong(40%), the type of enterprise was retail dealer(70.9%), and purchasing place was wholesale dealer from Seoul(51.8%). 7) And they thought that Yesan town market were too small(70.8%), and the diversifying of goods was not enough(35.5%). 8) The merchants of Yesan area showed that the degree of into venience to traffic system was 72.7% and its to parking facility was 83.3%. And they complained that community administration agency have not tried to activate market(79.1%). Based on the results of this study, the following recommendation were made for activating regional economy and community market. That is, it is to strengthen the economic function in community market so as to suffice the economic and social activities for settlement area as their living space. And it is necessary to improve the interrelationship of the central functions. 1) So, the accessibility between market center and its surrounding areas must be rised. ① the enforcing preferentially to pave the bus terminal and market center to link two place. ② the running of shuttle-bus two place every regular market day. ③ the widening and connecting the road to rise accessibility between market center and its surrounding area. ④ the enforcement of parking facility around market center. ⑤ the constructing marketing centered area as intergrated body of various marketing facilities in the adjacent site is the centered area planned to achieve efficient marketing. 2) The modernization and specialization market center facilities. 3) The strengthening of drawing surrounding area people into Yesan market center. So, the following recommended: ① the development of special products market ② inducing special projects ③ the development tour and leisure course drawing industrial facilities. 4) And it needs to enforce community people's local patriotism and to support community markets by administration agency.

신장이식 환자에서 HCMV 항원혈증 검사의 임상적 유용성

김탁,성흥섭,박관태,김송철,김성한,최상호,김양수,우준희,박수길,한덕종,이상오 대한감염학회 2009 감염과 화학요법 Vol.41 No.2

Background : This study was performed to determine the cut-off value and the predictability of symptomatic human cytomegalovirus (HCMV) infection according to the peak value of HCMV antigenemia assay in kidney transplant recipients. Materials and Methods : We reviewed the results of HCMV antigenemia assay (Chemicon, CA, USA) in patients who received kidney transplantation at our institution from May 2003 through May 2008, and investigated the existence and the type of HCMV infection by the medical record review. Patients who underwent the test only once during the episode or those who received ganciclovir for more than 48hrs before the test were excluded. The receiver-operator characteristic curve was drawn and the point showing maximum likelihood ratio (LR) was chosen as the cut-off value of symptomatic HCMV infection. Results : A total of 689 episodes were screened and 134 episodes were enrolled. Thirty-three (24.6%) episodes were symptomatic HCMV infection, 23 (17.2%) episodes were associated with HCMV syndrome, and 10 (7.5%) episodes were tissue-invasive diseases. The maximum LR was 7.5 (95% confidence interval, 4.014.2) and the cut-off value was 29.5 cells/200,000 WBC. The sensitivity, specificity, Positive predictive value, and negative predictive value were 66.7%, 91.1%, 71.0%, and 89.3%, respectively. Conclusions : The cut-off value of symptomatic HCMV infection by the peak value of HCMV antigenemia assay in our study was similar with previous results, although the sensitivity was relatively low.

Gene cloning and utility phophorylation assay of a protein-fused substrate for a highly sensitive detection of cdc2 protein kinase using a radioisotope detection technique for the development of a protein biochip

Ko, Kyong-Cheol,Choi, Mi Hee,Park, Sang Hyun John Wiley Sons, Ltd. 2009 Journal of labelled compounds & radiopharmaceutica Vol.52 No.4

<P>The prototype of the cdc2 protein kinase in mammalian cells regulates its entry into mitosis by phosphorylating a group of key proteins in the major cell cycle transitions. In this study, using the mep45 gene encoding the 45 kDa major envelope protein (Mep45) of Selenomonas ruminantium, a rumen bacteria, a Mep45-fused substrate (PKTPKKAKKL-Mep45, MFS-cdc2) was cloned to detect the activity of cdc2 protein kinase. We report here on a strategy for the detection of a phosphorylation of a substrate catalyzed by cdc2 protein kinase by using a radioisotope detection technique. It is possible to constantly obtain a reasonable quantity of MFS-cdc2 for the cdc2 protein kinase assay and its cost can be as low as a synthesized peptide. Results of the study indicate that the Mep45-fused protein can be used effectively as a substrate for detecting the activity of cdc2 protein kinase and it can be used in developing a protein biochip for a high-throughput screening and also for studying protein–protein interactions. Copyright © 2009 John Wiley & Sons, Ltd.</P> <B>Graphic Abstract</B> <P>Gene cloning and utility phophorylation assay of a protein-fused substrate for a highly sensitive detection of cdc2 protein kinase using a radioisotope detection technique for the development of a protein biochip Kyong-Cheol Ko, Mi Hee Choi, Sang Hyun Park<SUP>*</SUP>Using the mep45 gene encoding the 45 kDa major envelope protein (Mep45) of Selenomonas ruminantium, a rumen bacteria, a Mep45-fused substrate (PKTPKKAKKL-Mep45, MFS-cdc2) was cloned to detect the activity of cdc2 protein kinase. We report here on a strategy for the detection of a phosphorylation of a substrate catalyzed by cdc2 protein kinase by using a radioisotope detection technique. Copyright © 2009 John Wiley & Sons, Ltd. <img src='wiley_img/03624803-2009-52-4-JLCR1579-gra001.gif' alt='wiley_img/03624803-2009-52-4-JLCR1579-gra001'> </P>

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).

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).

Distribution of the Fairy Pitta (Pitta nympha) in the South Korea: A focus on protected areas

Gil-Pyo Hong(홍길표),Jae-Hoon Kim(김재훈),Seul-Gi Seo(서슬기),So-Yeon Cho(조소연),Bo-Yeon Hwang(황보연),Jong-Hyun Park(박종현),Seung-Yeon Lee(이승연),Ga-Young Kim(김가영),Wan-Hee Nam(남완희),Ha-Cheol Sung(성하철) 한국조류학회 2021 한국조류학회지 Vol.28 No.2

국내 팔색조(Pitta nympha) 분포 현황을 알아보고자 전국 국립공원 등 보호지역 중심으로 총 19개 지역을 조사하였고, 전문가 네트워크를 통해 자료를 보강하였다. 전국 19개 지역을 조사한 결과 북한산, 지리산 등 내륙지역 7개 국립공원에서는 팔색조가 확인되지 않았고, 경남 거제, 남해, 해남, 장흥 고흥 등 남해안과 인접한 12개 지역에서 팔색조 서식이 확인되었다. 또한 전문가 네트워크를 통해 수집한 팔색조 출현 위치 정보 82건을 분석한 결과, 전국 8개 광역지자체 모두 팔색조 서식이 확인되었고, 내륙지역(26.8%)보다 해안지역(73.2%)에서 더 많이 출현하였다. 연구 결과를 종합해보면, 우리나라의 팔색조는 주로 제주도와 남해안 지역을 중심으로 많이 분포하며, 그 보다 적은 개체수가 내륙 지역에 넓게 분포하고 있는 것으로 나타났다. The present study investigated the current distribution of Fairy Pitta (Pitta nympha) in South Korea. A total of 19 study areas, including protected areas such as national and provincial parks in South Korea were surveyed, and additional data obtained from an expert network. According to the results, Fairy Pitta was absent in seven inland national parks, including Mt. Bukhan and Chiri parks. Fairy Pitta inhabited 12 areas near the southern coast of South Korea, including Geoje, Namhae, Haenam, Jangheung, Goheung, and Gyeongnam Province. The analysis results of 82 datasets obtained from experts with regard to the locations where Fairy Pitta have been spotted showed that the bird inhabits all the eight metropolitan regions in South Korea, and is more frequently spotted in coastal areas (73.2%) than in inland areas (26.8%). The findings demonstrate that Fairy Pitta is mainly distributed in Jeju Island and the southern coast of South Korea, and is distributed in inland areas at relatively lower numbers.

인슐린비의존형 당뇨병 환자에서 심전도상 QT 간격 변수들과 허혈성 심장질환 발생의 관계

서승오,진도균,박진철,진희종,임성희,유형준,박성우,박종태 대한노인병학회 2000 Annals of geriatric medicine and research Vol.4 No.3