백서 뇌에서 전기경련 충격에 의해 발현이 조절되는 새로운 유전자

정선주,박정은,강웅구,구영진,박주배,김용식 大韓神經精神醫學會 2000 신경정신의학 Vol.39 No.5

연구목적: 이 연구는 백서 뇌에서 전기경련 충격(Electroconvulsive shock, ECS)에 의해 발현이 조절되는 새로운 유전자를 검색하고, 발달단게 및 뇌 조직에 따른 발현양상을 관찰하기 위하여 시행되었다. 방법: 생후 7일, 21일된 어린 웅성 백서에 전기경련 충격을 가한 후 mRNS differential display-PCR(DD-PCR) 기법을 시행하여 발현의 차이를 나타내는 유전자를 검색한 후 서열분석을 실시하였다. 생후 7일 및 21일된 백서의 뇌와 성년 백서의 대뇌피질, 해마를 이용한 Northern blot 분석을 시행하여 검색된 유전자의 ECS 처치 후 시간경과에 따른 유전자 발현양상을 관찰하였다. 결과: ECS 처치군과 위충격군간에 발현의 차이를 나타낸 유전자 4개를 검색할 수 있었으며, 이중 ECS에 의해 발현이 증가되는 1개의 유전자를 선정하여 서열분석을 실시한 결과 EST 서열만이 보고된 새로운 유전자임이 밝혀졌다. 이 유전자는 생후 7일된 백서의 뇌에서는 ECS 처치 후 180분에 발현양의 증가가 최고치에 도달하였으나, 생후 21일된 백서의 뇌와 성년백서의 대뇌피질 및 해마에서는 360분 경과시까지 지속적으로 발현 양이 증가하였다. 결론: 이같은 결과는 ECS에 의해 생체 내에서 유전자 발현의 조절이 이루어짐을 시사하며, 향후 이러한 유전자의 세포 내 기능에 관한 연구를 시행함으로써, ECS에 의한 생체의 반응과 치료기전에 대한 이해를 넓힐 수 있을 것으로 기대된다. 중심단어:전기경련 충격·DD-PCR·유전자·백서 뇌. Objectives: This study was performed to identify genes regulated by electroconvulsive shock (ECS) and to observe the pattern of expression of genes according to different developmental stages and brain regions. Method: ECS(130V, 0.5 sec) was given to male Sprague-Dawley rats with age of postnatal day 7 and 21(P7, P21 respectively). After screening genes regulated by ECS with mRNA diffe-rential display-PCR(DD-PCR), we selected one clone among them and observed the induction of this gene after ECS by time-dependent Northern blot analysis of rat brain of P7, P21 and adult rat cortex and hippocampus. Results: By DD-PCR method, we have identified four clones whose expression was regulated by ECS. Among them, one(CP 10-2) was proved to be a new gene by sequencing and BLAST search. Its expression was increased after ECS in P7, P21, and adult rat brain. The expression of CP 10-2 reached peak level at 180 minutes after ECS in P7 rat brain, but was further increased until 360 minutes after ECS in P21 and adult rat brain. Conclusion: In this study, a new gene was identified in rat brain which showed up-regulated expression in response to ECS. Cloning and characterization of this new gene would be helpful to elucidate the effect of ECS in rat brain. KEY WORDS:Electroconvulsive shock·Differential display-PCR·Gene·Rat brain.

인슐린 비의존형 당뇨병환자에 있어서 Multitestⓚ Kit 를 이용한 면역반응에 관한 연구

조정구,이용구 圓光大學校 1987 論文集 Vol.21 No.2

인슐린 비의존형 당뇨병환자에서 세포성 면역반응 및 감염과의 관련성 유무를 알아 보고져 1986년 1월부터 1986년 6월까지 원광대학병원 내과에 입원한 당뇨병환자 25명을 대상으로 Multitest CMI 방법을 이용하여 지연성 피부과민 반응을 측정 비교한바 정상 대조군과 환자군에 있어서 면역반응의 평균 득점치는 대조군 16.9 ±8.41 ㎜, 환자군 7.7±4.24 ㎜로 환자군에서 대조군에 비해 현저히 저하된 면역반응 ( P < 0.005) 을 보였으며, 환자군에서 면역반응과 당화 혈색소치와의 관련성은 유의한 음의 상관관계 (r = -0.50, P < 0.05) 를 보였으며 환자군중 감염이 있는 군의 면역반응 평균 득점치는 5.5 ±3.55 ㎜ , 비감염군은 9.4 ±4.02 ㎜로 감염군에서 유의한 감소 ( P<0.01)를 보였다. 결론적으로 Multitest CMI 방법은 편리하게 환자의 세포성 면역반응을 알아볼 수 있었으며, 혈당을 정상으로 유지하므로서 면역능을 강화하고 감염을 방지할 수 있음을 시사하였다. To assess cell mediated immunity in NIDDM, 25 NIDDM patients were evaluated by Multitest device. Delayed cutaneous hypersensitivity (DCH) responsiveness is quantified by a score ( the sum of the diameters of induration)at 48 hours. The results were summarized as follows 1) Compaired to male, significantly lower DCH response were observed in female of the control group ( P < 0.005 ), for the average scores (22.0 ㎜ for male, 10.8 for female). 2) In all ages classes, the patients had an average score significantly lower than that of the control subject(16.9 ㎜ for controls, 7.7 ㎜ for patients. P<0.005). 3) In the oldest group of the control subjects, DCH responses were decreased (8.4 ㎜) 4) There were significant cerrelations between DCH responses and glycosylated hemoglobins in diabetics ( r = -0.50, P < 0.05). 5) Compaired to patients without infection, Patients with infection had significant reductions in response to skin test (9.4 ㎜ for patients without infection, 5.5 ㎜ for patients with infection. P<0.01) 6) As the complications, no adverse reation was observed in both groups. Local reactions such as itching sensation and vesicle formation particularly to the tuberculin antigen were obsered 7 cases for the control groups and 2 cases for the patients. But these reactions disappeared within 8 to days without secondary infection or scar formation. In conclusion, the Multitest system appears to be safe, rapid and reproducible means of evaluating DCH and it suggested that better diabetic control may have a role in bolstering or augmenting cell mediated immunity and protecting against infections.

칼만증후군(Kallmann's Syndrome) 1례

김용성,백승훈,유경훈,구기선,형근영,김경년,조정구 圓光大學校 醫科學硏究所 1997 圓光醫科學 Vol.13 No.1-2

Kallmann's syndrome is the most common form of isolated gonadotropin deficiency, characterized by hypogonadotropic hypogonadism due to GnRH deficiency, delayed puberty and smelling difficulty. It occurs sporadic or familial pattern, and the mode of inheritence has not been fully documented. The defect in patient of Kallmann's syndrome occurs at suprapituitary level involving mechanism that regulate GnRH synthesis or release, so this syndrome classified as a secondary hypogonadotropic hypogonadism. The gonadotropin or pulsatile GnRH administration enable successful stimulation of spermatogenesis and fertility. We have experienced 1 patient with Kallmann's syndrome and presented with the review of the literature.

미립자 응집반응을 이용한 C-reactive Protein의 면역측정법에 관한 연구

최용경,정태화,최명자,김재화,최인성,김용호,송은영,이희구 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.1

환자의 복수와 늑막액으로부터 p-diazonium phenylphosphorylcholine(DPPC) coupled Separose-4B affinity chromatography와 hydroxylapatite chromatography를 실시하여 C-reactive protein (CRP)를 분리, 정제하였다. 정제된 CRP를 토끼에게 면역화하여 항혈청을 얻고 affinity chromatography를 하여 면역항체(IgG)를 분리하였다. 분리된 면역항체를 미립자에 감작시킨 후 미립자 응집반응에 의하여 3분내에 CRP를 측정할 수 있는 간이 면역측정법을 개발하였다. 본 연구에서 개발된 CRP측정법의 검출범위는 0.5∼20㎎/㎗이며, 임상 시험 결과 0.7∼2.9㎎/㎗에서는 강한 응집반응을, 5.0∼13.2㎎/㎗에서는 약한 응집반응을 보였고 28㎎/㎗이상에서는 항원 과잉으로 인한(zone of Ag excess phenomenon) 위음성을 나타냈다. 74명의 환자 혈청을 대상으로 CRP의 농도를 조사한 결과 평균치는 3.8㎎/㎗이었으며 대부분의 환자에서는 10㎎/㎗ 이하의 농도로 존재하였다. 그러므로 1차판정시 음성을 나타낸 시료라도 혈청을 5∼10배정도 희석하여 재분석한다면 오차없이 CRP를 검출할 수 있었다. 환자 혈청을 검체로 하여 본 연구에서 개발한 면역측정법과 현재 수입 시판중인 프랑스의 B사 제품과 일본의 I사 제품을 비교한 결과 좋은 상관관계를 보였다. 이와 같은 평가 분석을 통하여 볼 때 본 연구에서 개발한 간이 면역측정법은 사용이 비교적 간편하며 신빙성이 있어 CRP를 스크리닝 하는데 효과적임을 알 수 있었다. The C-reactive protein(CRP) from ascitic and pleural fluid was purified using calcium dependent affinity chromatography of CNBr activated Sepharose-4B covalently coupled to p-diazonium phenylphosphorylcholine(DPPC) and hydroxylapitite chromatography. Polyclonal antibody was prepared from rabbit by immunizing the purified CRP. Specific immunoglobulin G was isolated using affinity chromatography and coupled to microparticles. A sensitive microparticle-based immunoassay was developed to measure CRP within 3 mins. The detection range was between 0.5㎎/㎗ and 20㎎/㎗ in serum, showing strong response in the range of 0.7∼2.9㎎/㎗, week response in 5.0∼13.2㎎/㎗ and zone phenomenon over 28㎎/㎗. The average value of CRP in 74 samples was 3.8㎎/㎗ and most of the values were lower than 10㎎/㎗. The CRP values of serum samples were determined by our microparticle-based immunoassay, and were compared with those obtained using the other commercial products(B Co., France and I Co., Japan). Good correlations were shown between the values obtained by our developed microparticle-based immunoassay system and those by other commercial products. All performance characteristics evaluated make our developed microparticles-based immunoassay suitable for a simple, rapid, and reliable screening of CRP in serum.

합성 펩티드를 이용한 Interleukin-2수용체 β-Chain에 대한 단일클론항체의 제작

이희구,이홍수,이인애,최용경,최인성,정태화,김길현 이화여자대학교 생명과학연구소 1995 생명과학연구논문집 Vol.6 No.-

Hybridoma producing monoclonal antibodies reactive with the β-chain of interleukin-2 receptor (IL-2R) were generated from mice immunized with a synthetic peptide of 19 amino acids. The synthetic peptide constitutes strongly hydrophilic region of extracellular domain of the IL-2R molecules(residues 32-50) as revealed by hydrophathy plot analysis. Hybridomas were screened by ELISA using Hut102 cells and the synthetic peptide as antigenic moiety, and by miasuring the binding activity of antivodies to the Hut102 cell surface that was monitored by a flow cytometer. The antibodies were further screened by their inhibitory effect against the interaction between IL-2R and interleukin-2(IL-2). Antibodies were further characterized by immunoprecipitation followed by autoradiography to reveal that the antibodies recongnize the 75 KD protein molecules expressed on the Hut102 cell surface and human peripheral blood mononuclear cells(PBMC) shich confirms the antigenic moiety to be β-chain of IL-2 recepter molecules.

림프구성 뇌하수체염 - 증 례 보 고 -

임동준,정용구,이훈갑,이기찬,서중근 대한신경외과학회 1998 Journal of Korean neurosurgical society Vol.27 No.11

합성 펩티드를 이용한 Interleukin-2수용체 β-Chain에 대한 단일클론항체의 제작

이희구,이홍수,이인애,최용경,최인성,정태화,김길현 梨花女子大學校 藥學硏究所 1995 藥學硏究論文集 Vol.- No.5

Hybridoma producing monoclonal antibodies reactive with the ß-chain of interleukin-2 receptor (IL-2R) were generated from mice immunized with a synthetic pepride of 19 amino acids. The synthetic peptide constitutes strongly hydrophilic region of extracellular domain of the IL-2R molecules(residues 32-50) as revealed by hydrophathy plot analysis. Hybridomas were screened by ELISA using Hut102 cells and the synthetic peptide as antigenic moiety, and by measuring the binding activity of antibodies to the Hut102 cell surface that was monitored by a flow cytometer. The antibodies were further screened by their inhibitory effect against the interaction between IL-2R and interleukin-2(IL-2). Antibodies were further characterized by immunoprecipitation followed by autoradiography to reveal that the antibodies recongnize the 75 KD protein molecules expressed on the Hut 102 cell surface and human peripheral blood mononuclear cells(PBMC) which confirms the antigenic moiety to be ß-chain of IL-2 recepter molecules.

흰쥐 뇌에서 전기경련 충격에 의한 CREB 인산화의 발달단계에 따른 변화

강웅구,정희연,안용민,정선주,전송희,박주배,조수철,김용식 大韓神經精神醫學會 1999 신경정신의학 Vol.38 No.3

연구목적 : 이 논문에서는 뇌에서 전기경련 충격(Electroconvulsive shock. ECS)에 의한 전사인자 CREB의 인산화를 발달단계에 따라 알아봄으로써 정신장애의 신경발달학적 이해를 위한 생물학적 기반지식을 얻고자 하였다. 방 법 : 생후 7. 14. 21일 및 성년 흰쥐에 ECS를 가하고 시간별로 해마 및 소뇌 조직을 얻어 CREB의 인산화를 알아보기 위해 특이 항체로 면역블롯을 실시하였다. 결 과 : 발달단계에 따라 해마에서는 CREB이 감소한 반면 소뇌에서는 CREB이 증가하였다. 기저상태의 CREB 인산화는 해마와 소뇌에서 생후 7일에 비해 14일 이후 증가하였는데, 소뇌의 경우 CREB의 양증가와 비례하였다. ECS 후 CREB인산화 증가는 해마에서는 생후 21일 이후에 나타났으나, 소뇌에서는 생후 7일은 물론 성년에서도 나타나지 않았다. 결 론 : CREB 매개 신호전달은 발달단계별 및 조직별 활성의 차이를 나타내었으며 해마에서는 생후 21일 이후 ECS에 의해 활성화되었지만, 소뇌에서는 그렇지 않았다. 해마에서 ECS에 의한 CREB 인산화 증가는 c-fos 유전자의 발현과 관계있으리라고 생각되지만, CREB의 Ser-133 인산화 만으로는 발달단계 및 조직에 따른 c-fos 발현의 특이성을 설명할 수 없었다. Objectives : In order to understand the biological basis of neurodevelopmental perspectives of mental disorders, the authors investigated the developmental and regional changes in the phosphorylation of the transcription factor CREB following the electroconvulsive shock(ECS) in rat brain. Methods : Rats of various age groups(7, 14, 21 days postnatal and adults) were given ECS and their hippocampi and cerebella were dissected at specified time points. The content of CREB and phosphorylated CREB were measured by immunoblot analysis. Results : The amount of CREB increased in the hippocampus and decreased in the cerebellum according to the age. Baseline levels of CREB phosphorylation in both tissues were increased from postnatal 14 days, and it was proportional to the amount of CREB protein in the cerebellum. In the hippocampus, ECS increased the phosphorylation of CREB at postnatal 21 days, but in the cerebellum, ECS did not increased the phosphorylation of CREB in any age group. Conclusion : CREB mediated signal transduction pathways showed developmental and tissue-specific changes. ECS increased the phosphorylation of CREB in the hippocampus by postnatal 21 days, but not in the cerebellum. CREB activation is supposed to be related with the induction of c-fos after ECS in the hippocampus. However, the Ser-133 phosphorylation of CREB could not completely explain the developmental and tissue specificity of c-fos induction.

말단 비대증의 임상적 특성

박은미,안용환,김현정,김병수,임종주,김하영,박병현,조정구 圓光大學校 醫科學硏究所 2004 圓光醫科學 Vol.19 No.2

성장호르몬과 프로락틴을 분비하는 뇌하수체 선종이 동반된 Albright증후군 1예

오효정,김용성,신진호,김하영,박병현,조정구 圓光大學校 醫科學硏究所 2005 圓光醫科學 Vol.20 No.1