Nuclear factor I-C가 치근발생 과정에서 Hertwig's 상피초 형성에 미치는 영향

신인철,박주철,정문진,오현주,박선화,이창섭,김흥중,Shin, In-Cheol,Park, Joo-Cheol,Jeong, Moon-Jin,Oh, Hyun-Ju,Park, Sun-Hwa,Lee, Chang-Seop,Kim, Heung-Joong 대한소아치과학회 2005 大韓小兒齒科學會誌 Vol.32 No.3

치아의 형성은 상피-간엽간의 상호작용을 통해 조절되어지는 복잡한 발생과정이다. 지금까지 치관의 발생에 관여하는 유전자 및 그들의 신호전달경로에 관한 연구는 다수 진행되어 왔지만 치근의 발생을 조절하는 기전에 대해서는 별로 알려진 것이 없다. 최근에 NFI-C knock out 생쥐에서 정상치관에 비정상적인 치근을 가지는 치아가 보고되었다. 본 연구의 목적은 NFI-C가 어떻게 치근의 형태와 상아모세포의 분화에 관여하는지를 규명하는 것이다. NFI-C knock out 생쥐의 치근 발생동안에 HERS의 역할을 연구하고자 cytokeratin 면역조직화학적방법과 치근상아질의 특성을 규명하기 위해 DSPP mRNA in-situ hybrydization법을 수행하였다. 1. NFI-C knock out 생쥐의 치근형성시 HERS의 역할 Wild type과 knock out type 모두에서 cytokeratin은 모든 HERS 세포들과 반응하였고, HERS와 법랑상피 사이의 양성반응세포들의 연속성은 치경부 부위에서 소실되었다. Knock out type에서 치근상아질이 침착된 후, cytokeratin 양성-HERS 세포들은 치경부에서 불규칙한 배열과 극성의 상실을 보였다. 2. NFI-C knock out 생쥐의 치근상아질의 특성 DSPP mRNA의 발현은 wild type에서 치관과 치근상아질의 상아모세포 모두에서 강한 발현을 보인 반면, knock out type에서는 치관부위 상아질의 상아모세포에서만 강한 발현을 보였다. 3. NFI-C knock out 생쥐의 치근 발생과정에서 HERS는 치관으로부터 정상적인 확장을 보인 반면, 치근부위에서의 상아 모세포 분화는 실패하였다. 위의 결과들로 보아 NFI-C는 치근형성 과정에서 상아모세포 분화에 중요한 역할을 하는 것으로 사료된다. Tooth formation is a complex developmental process that is mediated through a series of reciprocal epithelial-mesenchymal interactions. Several signal pathways and transcription factors have been implicated in regulating molar crown development, but relatively little is known about the regulation of root development. It was reported that NFI-C knockout mice showed abnormal root formation with normal crown. The aims of this study are to elucidate how the NFI-C regulate the determine of root shape and odontoblasts differentiation. We carried out immunohistochemistry using cytokeratin to investigate the role of Hertwig's epithelial root sheath and DSPP mRNA in-situ hybridization to conform the nature of root dentin during root development in NFI-C knockout mice. Cytokeratin reacted with all the HERS cells and the continuity of cytokeratin positive cells between the HERS cells and enamel epithelium was lost in the cervical region both wild and K/O types. After root dentin deposition cytokeratin positive-HERS cells showed irregularity and loss of polarity in the cervical region in K/O type. DSPP mRNA was strongly expressed in odontoblasts of crown and root dentin in wild type mice, whereas expression of DSPP mRNA was restricted in odontoblast of crown dentin in the K/O type. During root formation in NFI-C knockout mice, HERS normally grow out of the crown but fail to induce odontoblast differentiation in root portion. These results suggest that NFI-C may play important roles in odontoblast differentiation during root dentin formation.

절제된 정상위벽의 자기공명영상기법에 관한 연구:조직소견과의 비교

서보경,설혜영,이남준,차인호,정규병,김정혁,박철민,이지영,Seo, Bo-Gyeong,Seol, Hye-Yeong,Lee, Nam-Jun,Cha, In-Ho,Jeong, Gyu-Byeong,Kim, Jeong-Hyeok,Park, Cheol-Min,Lee, Ji-Yeong 대한영상의학회 2001 대한영상의학회지 Vol.45 No.5

목적:급속자기공명영상기법을 이용하여 절제된 정상위벽을 관찰하고,이를 조직소견과 비교 연구하여,위벽의 관찰을 위한 최적기법을 찾고자 하였다. 대상과 방법:25명 환자의 모두 41개 절제된 정상위조직을 수술후 수지(polyethylene)통에 넣고,생리식염수로 채워 자기공명영상을 시행하였다.T1강조FLASH,지방억제T1강조FLASH, T2강조TSE와 True-FISP 등 4가지 기법으로 위조직의 영상을 얻었다.자기공명영상에서는 위벽의 층수와 각층의 신호강도를 관찰하였고,이를 조직소견과 비교하였다.자기공명영상소견을 조직소견과 비교한 후 위벽 각층의 명확성과 각층간의 구분,및 전체 영상의 질에 대하여 비교하였다.4가지 기법 중 가장 좋은 방법은 3,가장 나쁜 방법은 0으로 하여 등급을 판정하였다. 결과:자기공명영상에서 위벽의 층수는 T1강조FLASH에서 2층이 41예 중 6예(14.6%),3층 31예(75.6%),및 4층 4예(9.8%)였고,지방억제T1강조FLASH에서 2층 6예(14.6%)와 3층 35예(85.4%),T2강조TSE에서 3층 24예(58.5%),4층 11예(26.8%),및 5층 6예(14.6%)였으며,True-FISP에서 1층 2예(4.9%),2층 8예(19.5%),3층 23예(56.0%),4층은 4예(9.8%), 및 5층 4예(9.8%)이었다.위벽의 신호강도는 T1강조FLASH와 지방억제T1강조FLASH에서 2층으로 보인 예는 위내강으로부터 고-중등도,3층인 경우는 고-저-고/중등도,4층인 예는 고-저-고-중등도신호강도였다.T2강조TSE에서는 3층으로 보인 예는 등도/고-저-중등도,4 층인 경우는 중등도-저-고-중등도/저,5층인 예는 저-고-저-고-저신호강도였다.자기공명 영상소견을 조직소견과 비교하였을 때 위벽이 3층으로 보인 경우 이것은 “점막층-점막하층-근층 ”에 해당하였다.관찰한 3가지 면 모두에서 T1강조FLASH,지방억제T1강조FLASH,T2강조TSE 기법이 True-FISP보다 통계적으로 유의하게 우수하였다(p=0.001).점막층의 명확성에 있어서 가장 우수한 기법은 T1강조FLASH와 지방억제T1강조FLASH이었고(p<0.05),점막하층의 명확성과 점막하층과 근층간의 구분은 T2강조TSE가 가장 우수하였다(p<0.05).전체적인 영상의 질은 T1강조FLASH와 T2강조TSE에서 가장 우수하였다(p<0.05). 결론:자기공명영상은 위벽의 각 층을 구별할 수 있는 우수한 검사로 조직소견과 높은 연관성을 보이며,전체 영상의 질,점막하층의 명확성 및 점막하층과 근층간의 구분이 T2강조TSE에서 가장 우수한다. Purpose: To evaluate normal human gastric wall layers in vitro using magnetic resonance*(MR) imaging, to correlate the results with the histologic findings, and to determine the optimal technique for evaluation of the gastric wall. Materials and Methods: Forty-one normal resected gastric specimens obtained from 25 patients were dissected and placed in a polyethylene tube filled with normal saline. MR imaging with four MR sequences, T1-weighted FLASH*(T1FLASH), fat-saturated T1-weighted FLASH, T2-weighted TSE*(T2TSE), and True-FISP, was performed. The number of gastric wall layers and signal intensity of each layer were determined, and after correlating MR images with the histologic findings, the conspicuity of each layer*(mucosa, submucosa, and muscle), the distinction between each layer, and overall image quality were assessed. results: The gastric wall was shown by TIFLASH to have two (n=6, 14-6%), three (n=31, 75.6%) and four layers (n=4, 9.8%); by fat-saturated TIFLASH to have two (n=6, 14.6%) and three (n=35, 85.4%) ; by T2TSE to have three (n=24, 58.5%), four (n=11, 26.8%), and five (n=6, 14.6%); and by True-FISP to have one (n=2, 4.9%), two (n=8, 19.5%), three (n=23, 56%), four (n=4, 9.8%), and five (n=4, 9.8%) . The signal intensity of each layer at T1FLASH and fat-saturated T1FLASH was high-intermediate from the lumen in two-layer cases, high-low-high/intermediate in three-layer cases, and high-low-high-intermediate in four-layer cases. The signal intensity of each layer at T2TSE was intermediate/high-low-intermediate in three-layer cases, intermediatelow-high-intermediate/low in four-layer cases, and low-high-low-high-low in five-layer cases. Three-layered gastric wall corresponded mostly to mucosa, submucosa, and muscle from the inner to outer layers, respectively. T1FLASH, fat-saturated T1FLASH, and T2TSE were superior to True-FISP in evaluating the gastric wall. T1FLASH and fat-saturated T1FLASH were the best sequences for demonstrating mucosa (p<0.05), and T2TSE was the best for submucosa and the distinction between this and muscle (p<0.05). Both T1FLASH and T2TSE provided the best overall image quality (p<0.05). Conclusion: In-vitro MR imaging is an excellent technique for the evaluation of layers of normal gastric wall. T2TSE is the sequence which best demonstrates the conspicuity of submucosa, the distinction between submucosa and muscle, and overall image quality.

법랑모세포 분화와 성숙과정에서 OD314의 발현

박주철,안성민,김흥중,정문진,박민주,신인철,손호현 大韓齒科保存學會 2005 Restorative Dentistry & Endodontics Vol.30 No.5

법랑모세포는 법랑질을 형성하고 유지하는 세포로, 법랑질의 유기기질을 분비하고 법랑질 석회화 과정에도 관여한다. 치아 발생과정에서 법랑모세포의 분화는 순차적인 상피-간엽 상호작용에 의하여 조절되나, 분화나 성숙과정의 정확한 기전은 아직까지 잘 알려져 있지 않다. 최근에 상아모세포에서 처음 발견된 OD314가 치아 발생과정에서 상아질을 형성하는 상아모세포 뿐 아니라 법랑모세포에도 발현된다고 하였다. 이에 본 연구에서는 생쥐 하악 전치의 다양한 시기의 법랑모세포를 이용하여, 형태학적 분석과 in-situ hybridization에 의한 OD314 mRNA의 발현 그리고 OD314 항체를 이용한 면역조직화학적 분석을 통하여 OD314 유전자의 법랑 모세포 분화와 성숙과정에서의 역할을 연구하여 다음과 같은 결과를 얻었다. 1. 형태학적으로 법랑모세포는 분화 단계에 따라 분비 전단계 법랑모세포, 분비기 법랑모세포, 성숙기의 평탄끝 법랑모세포와 성숙기의 주름끝 법랑모세포로 구분되었다. 2. OD314 mRNA는 분비기의 법랑모세포에서부터 발현되기 시작하여 법랑모세포가 성숙해갈 수록 그 발현이 증가하였다. 3. OD314 단백질은 분비 전단계의 법랑모세포에서는 발현되지 않고, 분비기의 법랑모세포에서는 세포질에 전체적으로 발현되었다. 성숙기의 평탄끝 법랑모세포와 주름끝 법랑모세포에서는 세포의 근심과 원심끝단에 OD314 단백질이 강하게 발현되었다. 이상의 결과를 종합하여 OD314는 법랑모세포의 분화와 성숙과정에서 세포질 내부에서 특징적인 역할을 하는 것으로 사료된다. Ameloblasts are responsible for the formation and maintenance of enamel which is an epithelially derived protective covering for teeth. Ameloblast differentiation is controlled by sequential epithelial-mesenchymal interactions. However, little is known about the differentiation and maturation mechanisms. OD314 was firstly identifled from odontoblasts by subtraction between odontoblast/pulp cells and osteoblast/dental papilla cells, even though OD314 protein was also expressed in ameloblast during tooth formation. In this study, to better understand the biologcal function of OD314 during amelogenesis, we examined expression of the OD314 mRNA and protein in various stages of ameloblast differentiation using in-situ hybridization and immunohistochemistry. The results were as follows : 1. The ameloblast showed 4 main morphological and functional stages referred to as the presecretory, secretory, smooth-ended, and ruffle-ended. 2. ○D314 mRNA was expressed in secretory ameloblast and increased according to the maturation of the cells. 3. OD314 protein was not expressed in presecretory ameloblast but expressed in secretory ameloblast and maturative ameloblast. OD314 protein was distributed in entire cytoplasm of secretory ameloblast. However, OD314 was localized at the proxiamal and distal portion of the cytoplasm of smooth- ended and ruffle-ended ameloblast. These results suggest that ○D314 may play important roles in the ameloblast differentiation and maturation.

Qualitative Analysis of Proteins in Two Snake Venoms, Gloydius Blomhoffii and Agkistrodon Acutus

Su-Jeong Ha,Yeo-Ok Choi,Eun-Bin Kwag,Soo-Dam Kim,Hwa-Seung Yoo,In-Cheol Kang,So-Jung Park 대한약침학회 2022 Journal of pharmacopuncture Vol.25 No.1

Objectives: Snake venom is a complex mixture of various pharmacologically active substances, such as small proteins, peptides, and organic and mineral components. This paper aims to identify and analyse the proteins in common venomous snakes, such as Gloydius blomhoffii (G. blomhoffii) and Agkistrodon acutus (A. acutus), in Korea. Methods: We used mass spectrometry, electrophoresis, N-terminal sequencing and in-gel digestion to analyse the proteins in these two snake venoms. Results: We identified eight proteins in G. blomhoffii venom and four proteins in A. acutus venom. The proteins detected in G. blomhoffii and A. acutus venoms were phospholipase A2, snake venom metalloproteinase and cysteine-rich secretory protein. Snake C-type lectin (snaclec) was unique to A. acutus venom. Conclusion: These data will contribute to the current knowledge of proteins present in the venoms of viper snakes and provide useful information for investigating their therapeutic potential.

관상동맥 스텐트 시술 후의 재협착에 관한 연구

김윤철,이정우,김보영,강정아,임대승,이민수,김정희,성보영,최성준,성인환,전은석 충남대학교 의과대학 지역사회의학연구소 2000 충남의대잡지 Vol.27 No.1

Coronary stent implacement is known as an effective treatment in the intimal dissection after percutaneous transluminal coronary angioplasty and the prevention of restenosis. However, In-stent restenosis still remains a major concern in clinical stenting. The stents were placed in 103 patients from July 1996 to March 1999 and performed follow-up coronary angiograms in 59(57.3%) patients. To identify the clinical, angiographic and procedurerelated variables 'which predict late restenosis within the stented artery, 59 patients(58.3±9.9, M:F= 41:18) were studied. The clinical characteristics of the patients were stable angina in 23(39.0%), unstable angina in 14(23.7%), acute myocardial infarction in 21(35.6%) and old myocardial infarction in 1(1.7%). Coronary stenting was performed in 1 patient(1.7%) for primary lesion, 50 patients(84.7%) for suboptimal results after PTCA, 6 patients(10.2%) for bail-out procedure, and 2 patients(3.4%) for restenotic lesions. All patients were treated with aspirin and ticlopidinc. The follow-up angiograms were obtained at 7±4 months. The overall in-stent restenosis rate was 27.1%. The coronary angiographic findings were 32 single vessel(54.2%), 19 two vessel(32.2%) and 8 three vessel disease(13.6%). The angiographic morphological characteristics were type A in 33(55.9%), type B in 14(23.7%), type C in 12(20. 3%) cases. Variables of 16 patients with restenosis were compared with those of 43 patients without restenosis. Previously known predictors for in-stent restenosis were multiple stenting, stenting for restenotic lesions, residual stenosis after stenting, stenting for total occlusion lesions, reference diameter, balloon to vessel ratio, acute gain and minimal luminal diameter after procedure, design and characteristics of stents, ostial lesion of aorta, high pressure method for stenting, lesion length, diabetes mellitus, size of artheroma, saphenous vein grafts, ulcerlating lesions and calcified lesions. In this study, Reference diameter before stenting(2.43±0.54mm vs. 2.88±0.65mm, p=0.016) and balloon-to-artery ratio(1.28±0.26 vs. 1.11±0.18, p=0.006) were predictors for in-stent restenosis. 1) The overall in-stent restenosis rate was 27.1%. 2) In the analysis of predictors for in-stent restenosis, there was no significant differences in clinical, angiographic factors between group with restenosis and without restenosis. But, Only reference diameter before stenting and balloon-toartery ratio were predictors of late in-stent restenosis. In conclusion, stenting is effective revascularisation method for selected patients with ischemic heart disease, and to minimize in-stent restenosis rate, stent implanting is achieved in a large vessel on the basis of an artery-to-stnet ration of 1:1, if possible.

The Implication of Cardiac Injury Score on In-hospital Mortality of Coronavirus Disease 2019

Kim In-Cheol,Song Jin Eun,Lee Hee Jung,Park Jeong-Ho,Hyun Miri,Lee Ji Yeon,Kim Hyun Ah,Kwon Yong Shik,Park Jae Seok,윤종찬,Hwang Jongmin,Lee Cheol Hyun,Cho Yun-Kyeong,Park Hyoung-Seob,Yoon Hyuck-Jun,Nam 대한의학회 2020 Journal of Korean medical science Vol.35 No.39

Backgrounds: The severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) has spread worldwide. Cardiac injury after SARS-CoV-2 infection is a major concern. The present study investigated impact of the biomarkers indicating cardiac injury in coronavirus disease 2019 (COVID-19) on patients' outcomes. Methods: This study enrolled patients who were confirmed to have COVID-19 and admitted at a tertiary university referral hospital between February 19, 2020 and March 15, 2020. Cardiac injury was defined as an abnormality in one of the following result markers: 1) myocardial damage marker (creatine kinase-MB or troponin-I), 2) heart failure marker (N-terminal-pro B-type natriuretic peptide), and 3) electrical abnormality marker (electrocardiography). The relationship between each cardiac injury marker and mortality was evaluated. Survival analysis of mortality according to the scoring by numbers of cardiac injury markers was also performed. Results: A total of 38 patients with COVID-19 were enrolled. Twenty-two patients (57.9%) had at least one of cardiac injury markers. The patients with cardiac injuries were older (69.6 ± 14.9 vs. 58.6 ± 13.9 years old, P = 0.026), and were more male (59.1% vs. 18.8%, P = 0.013). They showed lower initial oxygen saturation (92.8 vs. 97.1%, P = 0.002) and a trend toward higher mortality (27.3 vs. 6.3%, P = 0.099). The increased number of cardiac injury markers was significantly related to a higher incidence of in-hospital mortality which was also evidenced by Kaplan-Meier survival analysis (P = 0.008). Conclusion: The increased number of cardiac injury markers is related to in-hospital mortality in patients with COVID-19.

저 Pb Sn-5%Pb-1.5%Ag-x%In계 솔도 합금의 특성에 관한 연구

홍순국,주철홍,강정윤,김인배,Hong, Sun-Guk,Ju, Cheol-Hong,Gang, Jeong-Yun,Kim, In-Bae 한국재료학회 1998 한국재료학회지 Vol.8 No.11

Pb의 환경오염 문제를 발생하지 않는 저농도 Pb 솔도합금을 개발하기 위하여, 새로운 Sn-5%Pb-1.5%Ag-x%In계 합금 조성을 설계하고, 이 합금의 융점, 젖음성, 상분석, 경도, 인장강도, 드로스성을 평가하여, Sn-37%Pb 솔더오 대체 가능성을 타진하였다. Sn-37%Pb 솔도 합금의 Pbdldhs 용출농도는 국제규제치인 3ppm보다 훨씬 적은 0.46ppm이었고, 환경문제를 유발하지 않는 것으로 확인되었다. 이 합금계의 융점은 $183-192^{\circ}C$이고, 응고온도범위도 $5^{\circ}C$내외로 매우 좁았다. 젖음성은 In의 첨가양에 따라 큰 차이가 거의 없었으며, Sn-375Pb와 비슷하였다. 융점 및 젖음성 측면에서 Sn-37%Pb와 대체 가능한 것으로 판단되었다. 경도는 Sn-37%Pb의 약 1.5배이고, 인장강도는 Sn-37%Pb의 것보다 높고, In의 첨가량에 따라 증가하였지만, 연신율은 감소하였다. In이 1% 첨가된 합금에서는 수지 상정 경계에 Ag3Sn과 Pb가 정출되고, 3% 이상에서는 $Ag_3Sn$과 $Ag_3In$ 및 Pb가 정출되었다. 드로스 생성속도는 Sn-37%Pb 합금이 Sn-5%Pb-1.5%Ag 합금보다 빠르고, In을 첨가할수록 느리고 2%의 In을 첨가한 합금은 180분에서도 거의 드로스가 발생하지 않았다. This work designed Sn-5%Pb-1.5%Ag-x%In solder alloy to develop the solder alloy with low Pb content. This solder alloy doesn't cause environmental pollution. and this study reviewed the probability of replacement of Sn-37%Pb solder as evaluation of melting range, wettability. microstructure, microhardne'ss, tensile strength, drossability of this new solder alloys. The level of international regulation in dissolution amount of Pb ion was 3ppm. But dissolution amount of Pb ion in Sn-5%Pb solder alloy confirmed not to threat the global environmental is 0.46ppm. The melting range of this solder alloy was $183-192^{\circ}C$. Also the range of solidification was very narrow within $5^{\circ}C$. The wettability was similar to Sn-37%Pb solder, and the effect of amount of In addition of wettability couldn't be founded. The probability of replacement in the melting range and wettability is very high. And microhardness of this solder alloy was 1.5 times of conventional type solder. Tensile strength of new solder alloys was a little high than that of conventional type solder. With increasing amount of In% addition, tensile strength was increased, but elongation was decreased. The solder alloy of l%In addition revealed AgSn and Pb on dendrite microstructure boundary, and $Ag_3Sn$, $Ag_3In$ and Pb were revealed on it at the solder alloy of 3% In addition. The drossability was superior to Sn-37%Pb solder alloy and the solder alloys of 2% In addition was not generated for 3hrs.

Clinical impact of admission hyperglycemia on in-hospital mortality in acute myocardial infarction patients

Kim, Eun Jung,Jeong, Myung Ho,Kim, Ju Han,Ahn, Tae Hoon,Seung, Ki Bae,Oh, Dong Joo,Kim, Hyo-Soo,Gwon, Hyeon Cheol,Seong, In Whan,Hwang, Kyung Kuk,Chae, Shung Chull,Kim, Kwon-Bae,Kim, Young Jo,Cha, Kwa Elsevier 2017 INTERNATIONAL JOURNAL OF CARDIOLOGY Vol.236 No.-

<P><B>Abstract</B></P> <P><B>Background</B></P> <P>Acute hyperglycemia on admission is common in acute myocardial infarction (AMI) patients regardless of diabetic status, and is known as one of prognostic factors. However, the effect of hyperglycemia on non-diabetic patients is still on debate.</P> <P><B>Methods</B></P> <P>A total of 12,625 AMI patients (64.0±12.6years, 26.1% female) who were enrolled in Korea Acute Myocardial Infarction Registry-National Institute of Health between November 2011 and December 2015, were classified into 4367 diabetes (65.4±11.6years, 30.4% female) and 8228 non-diabetes (63.3±13years, 23.9% female). Patients were analyzed for in-hospital clinical outcome according to admission hyperglycemic status.</P> <P><B>Results</B></P> <P>In diabetic patients, independent predictors of in-hospital mortality were old age, high HbA<SUB>1</SUB>C, pre-Thrombolysis In Myocardial Infarction (TIMI) flow 0, left ventricle ejection fraction<40%, cardiogenic shock and ventricular tachycardia. In non-diabetic patients, independent predictors of in-hospital mortality were old age, high admission glucose (≥200mg/dL), pre TIMI flow 0, failed percutaneous coronary intervention, low left ventricle ejection fraction<40%, cardiogenic shock, stent thrombosis and decreased Hb≥5g/dL. In hospital mortality was significantly higher in diabetic patients compared to non-diabetic patients (5.0% vs. 3.4%, <I>p</I> <0.001). However, non-diabetic patients with hyperglycemia have significantly higher mortality compared to diabetic patients (17.4% vs. 7.2%, <I>p</I> <0.001). Comorbidity including cardiogenic shock (<I>p</I> <0.001), cerebral hemorrhage (<I>p</I> =0.012), decreased Hb≥5g/dL (<I>p</I> =0.013), atrioventricular block (<I>p</I> <0.001) and ventricular tachycardia (<I>p</I> =0.007) was higher in non-diabetic with hyperglycemia than in diabetic patients.</P> <P><B>Conclusions</B></P> <P>These findings underscore clinical significance of admission hyperglycemia on in-hospital mortality in non-diabetic AMI patients.</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).

Optimal Timing of Percutaneous Coronary Intervention in Patients With Non–ST-Segment Elevation Myocardial Infarction Complicated by Acute Decompensated Heart Failure (from the Korea Acute Myocardial Infarction Registry-National Institutes of Healt

Kim, Min Chul,Jeong, Myung Ho,Sim, Doo Sun,Hong, Young Joon,Kim, Ju Han,Ahn, Youngkeun,Ahn, Tae Hoon,Seung, Ki Bae,Choi, Dong-Joo,Kim, Hyo-Soo,Gwon, Hyeon Cheol,Seong, In Whan,Hwang, Kyung Kuk,Chae, S Elsevier 2018 The American journal of cardiology Vol.121 No.11

<P>The optimal timing of percutaneous coronary intervention (PCI) in patients with non–ST-segment elevation myocardial infarction (NSTEMI), complicated by acute decompensated heart failure (ADHF), is unclear. A total of 1,027 patients with NSTEMI complicated by ADHF who underwent successful PCI were analyzed using a Korean multicenter registry. All patients were divided into 4 groups by the timing of PCI: group 1 (PCI < 2 hour after admission, n = 149), group 2 (2 to 24 hours, n = 577), group 3 (24 to 72 hours, n = 189), and group 4 (≥72 hours, n = 112). We analyzed the incidences of 12-month mortality, nonfatal myocardial infarction (MI), target-vessel revascularization, and rehospitalization because of HF. The prevalence of ADHF in patients with NSTEMI was 15.2% at initial presentation, and in-hospital mortality was higher in group 1 than in the other groups. There were no significant differences in mortality, nonfatal MI, target-vessel revascularization, or rehospitalization for HF during the 12-month follow-up between groups, regardless of initial PCI timing, except for a higher 12-month mortality in patients who received PCI within 24 hours (vs ≥24 hours) (hazard ratio 1.52, 95% confidence interval 1.09 to 2.29, p = 0.046). Early PCI did not reduce adverse clinical outcomes in patients with NSTEMI complicated by ADHF. Delayed PCI after stabilization may be reasonable in such high-risk patients.</P>