Global gene expression profiling of congestive heart failure subsequent to experimental left ventricular pressure overload pretreated with the peroxisome proliferator-activated receptor-gama agonist Rosiglitazone

高血壓一直為公衛上一個相當重要的議題。?所周知，高血壓是許多神經系統以及 心血管系統的併發症的一個重要的危險因子。好好控制高血壓可以有效增進患者的預 後。然而根據研究指出，仍然有三分之一的患者，其掏血壓仍無法被控制在滿意的範圍 之內。長期未良好控制的高血壓將會使左心室的心室後負荷上升，心肌細胞為了維持心 輸出量，心肌細胞會變的肥厚來應付所需的功，於是乎左心室肥厚便產生。而同時，纖 維母細胞也會被活化，使的collagen 產量增加，而使的間質增生亦隨之增加，並開始 產生心肌細胞之纖維化。早期只是單純的心室肥厚，因此雖有心臟舒張功能失調，但是 心肌之收縮功能正常；而到晚期，心臟的收縮功能亦會受到影響，而進展到代償不良之 心臟衰竭。心肌梗塞後，細胞骨架和收縮元件的變化，與Actin 和myosin 基因表現改 變相關，而Collagen 和fibronectin 基因表現則是梗塞組織纖維化的部份原因。梗塞 後，為了改善血行動力，心肌組織中atrial natriuretic peptide mRNA 及其蛋白質產 物也會增加；而內皮一氧化氮生成?(eNOS)，細胞外間質(ECM)metalloproteinases (MMPs)和腫瘤壞死因子(TNF)的基因表現亦牽涉其中，同時存活的心肌組織也會變的更 為肥厚。這些變化皆與心肌梗塞後心臟進行再塑型(remodeling)作用息息相關，也對梗 塞造成之心臟衰竭演進有相當之影響。 過氧化體增殖劑活化受器(PPARs)屬細胞核內受器群，共分三種亞型a 、b /d 、和g ， 其為轉譯因子，可調節基因表現並控制不同方面的細胞功能，包含脂肪和脂蛋白的代 謝，脂肪酸氧化，葡萄糖代謝，脂肪新生，細胞分化等等。近來研究亦發現PPARs 表現 也會影響免疫細胞、巨嗜細胞、內皮細胞和血管平滑肌細胞，並影響不同細胞之細胞激 素(cytokines)分泌或其他酵素分泌(如MMPs)，因此對血管硬化之調控亦有重要角色。 PPAR g 促進劑在動物模式顯示可減少心肌梗塞區域及收縮功能失常，且對於梗塞後左心 室再塑型作用及心臟衰竭也具有減緩之作用。不僅如此，對於心肌肥厚的進展，PPAR g 也在不同研究中顯示有減緩之作用。唯目前僅知道與部分細胞產物有關，對於長期左心 室壓力過度負荷後心肌整體影響以及保護機轉明瞭則相當少。 cDNA 微陣列晶片(cDNA microarray)可以同時分析成千的基因表現，對疾病致病機轉研 究助益甚大。本研究希望藉此有效的研究平台來深入解析PPAR g 促進劑Rosiglitazone 對 心肌梗塞可能具保護作用的機轉以及其影響梗塞後心衰竭演進的機制，並希望應用到日 後心肌梗塞之治療。Aim of study: To explore the gene expression profiles of congestive heart failure (CHF) subsequent to experimental pressure overload pretreated with peroxisome proliferator-activated receptor-g (PPARg) agonist Rosiglitazone. And to elucidate the potential benefits and mechanisms of PPARg agonist on pressure-overload hemodynamics, cardiac remodeling, and CHF. Background: Hypertension (HTN) continues to be of major public health concern. Increased risks of several adverse neurological and cardiovascular outcomes were documented in the population of HTN. Longstanding HTN increased the left ventricular (LV) afterload markedly, and cardiac fibrosis evolves as an adaptive response to hemodyanamic overload. Subsequently, interstitial fibrosis occurred and contributed to LV stiffening and impaired compliance. Although initially the LV systolic function is well preserved with simply diastolic dysfunction, however progression to decompenated stage congestive heart failure (CHF) with systolic dysfunction is hastened. PPARs are nuclear receptors, which function as transcription factors the regulate different genes expression. They control a variety of cellular functions and are involved in control of vascular inflammation, thrombogenecity and inflammatory cytokines. Several animal studies had shown that ligands of the PPARs can reduce infarct size and potentially protect the heart against ischemia-reperfusion injury. PPARg ligands have also been shown to suppress the development of cardiac myocyte hypertrophy both in vitro and in the in vivo setting. However, knowledge about the influences of PPARs on pressure-loaded CHF and cardiac remodeling process at cellular level is quite limited. cDNA microarray offers a genome-wide simultaneous analysis of gene expression profiling changes in perturbed physiological or pathological conditions. With the aid of this powerful tool, researches could progress efficiently. Experimental protocol: Rosiglitazone-pretreated or placebo-preteated rats undergo abdominal aorta ligation resulting in left ventricular overload. Two-color cDNA microarray system containing 7,600 clones from regional company is applied to explore the cardiac differential gene expressions between Rosiglitzaone-pretreated and placebo-pretreated rat pressure overload model. Serum biochemistry, hemodynamics, echocardiography, and clinical conditions are also measured prior to sacrifice and will be analyzed. Expected results: The two-color cDNA microarray data analysis will be validated utilizing RT-PCR. Through this study, Rosigliazone pretreatment effects, whether beneficial or not, on pressure overload cardiac remodeling and CHF course could be evaluated, and differential cardiac gene expressions contributing to the probable beneficial effect of Rosiglitazone on CHF subsequent to pressure overload could be elucidated

以雙色cDNA微陣列技術探討同半胱胺酸對血管內皮細胞之作用

本 計畫主要是建立基因微陣列(cDNA microarray)之數據分析篩選出差異表達基因，並 選出血管新生相關之基因群。在人體不易取得之腫瘤組織開始實驗及分析前，我們先以細胞 培養模式，使研究工作者熟習細胞培養技術，RNA 之抽取及cDNA 微陣列晶片之實驗技術，取 得影像資料後再加以統計分析，建立研究平台。 初 步研究重點為藉由cDNA 微陣列技術來探討在不同濃度及時間下同半胱胺酸 (homocysteine ，Hcy)對 培養血管內皮細胞的影響。因此，現階段著重分離臍帶靜脈血管內 皮細胞 ((HUVEC)技術之建立和細胞培養技術之穩定。我們可成功地從新生嬰兒的臍帶分離出 原始臍帶靜脈血管內皮細胞 ((primary cells)，以完全培養液在37 ℃及5 ％ CO2 培養箱中培 養，每隔二至三天，更換新的培養液。待細胞長滿後，繼代培養之。另外為了確定分離出的 細胞是否為血管內皮細胞，而非纖維母細胞 (fibroblasts)或是平滑肌細胞(smooth muscle cells)，我們先利用光學顯微鏡觀察細胞，培養的HUVEC 為同種的 ((homogenous)且 單層地 ((monolayer)貼附生長；細胞外形呈多邊形，具有典型〝cobblestone 〞外觀。同時也 擬利用immunocytochemistry 的方法來鑑定內皮細胞上特有的marker 蛋白質，如CD-31 、 factor VIII-related antigen ，佐證我們所培養的細胞為內皮細胞。除此之外，在測試Hcy 對HUVEC 細胞的作用之前，我們欲先取第二代至第四代的正常HUVEC 細胞，利用RNA pure Kit (Genesis 代理)萃取RNA ，檢查RNA 濃度及純度；若RNA 濃度或純度太低，考慮是否要增加細 胞數目或改良萃取方法。以估算未來從事cDNA microarray 所需的細胞數目及RNA 濃度，決 定要取多少細胞來進行Hcy 測試較為恰當

[[alternative]]Investigating the Junior High School Students' Concepts Regarding Chemical Change by Means of Group Demonstration Test and with a Particle-model-simulation Instruction: The Production of Gas as an Example

[[abstract]]In daily life, it is full of chemical change. However, because of ordinary terms, direct judgment, and improper teaching, students have quite a lot of wrong recognition of these changes which are normal phenomenon. Taking the chemical change of producing gas for example, this research was aimed at investigating students’ recognition of correlated concepts by means of group demonstration test using self-designed experimental instrument and the problems of two-tier diagnostic test. Furthermore, by means of the experiment-manipulating observation and particle-model-simulation instruction, students could improve their concepts and changed their misconceptions concerning the chemical changes of gas production. The subjects including all the grades in the junior high school, were divided into the experimental group and the control group. There were totally 424 students. The main findings in this research were: 1.Most students were unclear about the definition of chemical changes, lack building up the concept of material, and were not good at distinguishing the variety of changes. Moreover, most students were unclear about the process of chemical changes, and unable to think the character of microscopic particles. Therefore, they held misconceptions of chemical changes. No matter in the microscopic or macroscopic view, students’ main misconceptions were “making something out of nothing” and “disappear out of the void”. 2.Comparing the difference in the recognition of the concepts of chemical change, students’ recognition of the most concepts was progressed while their grades were higher. It implies that the comprehension of chemical changes needs students’ mature ability of abstract thinking. 3.Comparing the improvement between the experimental and the control group before and after the instruction, it proved that the experiment-manipulating observation and particle-model-simulation instruction were significantly effective in understanding the process and the particle concepts of chemical changes. Moreover, in this research, it showed that there was much effective for the eight grade students. It means that by model instruction it had better effectiveness for helping the students who were developing their ability of abstract thinking.

Association of Circulating Matrix Metalloproteinase-1, but Not Adiponectin, with Advanced Coronary Artery Disease

Objective: Recent evidence Suggests that high tissue matrix metalloproteinase-1 (MMP-1) and low adiponectin may serve as biomarkers of atherosclerosis. Results on the associations of circulating MMP-1 and adiponectin concentrations are scarce. We hypothesized that patients with multivessel coronary artery disease (CAD) have elevated high-sensitivity C -reactive protein (hs-CRP), MMP-1 but low adiponectin levels, and concomitant measurements of these biomarkers Could improve predictive strength for advanced CAD. Research design and methods: We analyzed concentrations of MMP-1, hs -CRP and adiponectin in 217 subjects with angiographically documented multivessel CAD (two-, or three-vessel disease by luminal stenosis >= 50%) and 81 controls. MMP-1 and hs-CRP were notably higher in patients with CAD: while adiponectin was not significantly different between two groups. Levels of hs-CRP positively correlated with body mass index and left ventricular dysfunction (R-2 = 0. 16, P<0.0001): while adiponectin was significantly associated with age, gender,and levels of cholesterol and triglyceride (R-2 = 0.09, P < 0. 0001 ). On the contrary, MMP-1 was not associated with any clinical cardiovascular risk factors, and still an independent predictor(OR=1.49, P <0.0001) of multivessel CAD after the adjustment of clinical risk factors and hs-CRP. Conclusion: Elevated MMP-1 and hs-CRP, but not low adiponectin concentrations, could predict the presence of advanced coronary atherosclerosis. In addition, MMP-1 may serve as a more specific market for significant CAD independent of hs-CRP

Increased Matrix Metalloproteinase-1 Concentrations Are Associated with Advanced Coronary Artery Disease

Recent evidence Suggests that high tissue matrix metalloproteinase-1 (MMP -1) and low adiponectin may serve as biomarkers of atherosclerosis. Results on the associations of circulating MMP-1 and adiponectin concentrations are scarce. We hypothesized that patients with multivessel coronary artery disease (CAD) have elevated high-sensitivity C-reactive protein (hs-CRP), MMP-1 but low adiponectin levels, and concomitant measurements of these biomarkers Could improve predictive strength for advanced CAD. Research design and methods: We analyzed concentrations of MMP-1, hs -CRP and adiponectin in 217 subjects with angiographically documented multivessel CAD (two-, or three-vessel disease by luminal stenosis >= 50%) and 81 controls. MMP-1 and hs-CRP were notably higher in patients with CAD: while adiponectin was not significantly different between two groups. Levels of hs-CRP positively correlated with body mass index and left ventricular dysfunction (R-2 = 0.16, P<0.0001): while adiponectin was significantly associated with age,gender,and levels of cholesterol and triglyceride (R-2 = 0.09, P < 0. 0001). On the contrary, MMP-1 was not associated with any clinical cardiovascular risk factors, and still an independent predictor(OR=1.49, P<0.0001) of multivessel CAD after the adjustment of clinical risk factors and hs-CRP. Conclusion: Elevated MMP-1 and hs-CRP, but not low adiponectin concentrations, could predict the presence of advanced coronary atherosclerosis. In addition, MMP-1 may serve as a more specific market for significant CAD independent of hs-CRP

經 食道超音波心圖應用時代的感染性心內膜炎診療

During a 45-month period, 50 consecutive patients with infective endocarditis were evaluated at the National Taiwan University Hospital with emphasis on the role of transesophageal echocardiography (TEE) in the management of these patients. Among them, rheumatic heart disease was still the most common underlying cardiac disorder (10/50, 20 %), while mitral valve prolapse (8/50, 16%) and congenital heart disease (8/50, 16%) were also frequently encountered. More than one third (19/50, 38%) had no underlying heart disease. Four intravenous drug abusers, quite rare previously in Taiwan, were found during the study period. Native valves involved were mostly mitral valve (n = 18), aortic valve (n = 15), and both mitral and aortic valves (n= 3). Tricuspid valve and pulmonic valve were involved in 3 and 2 patients, respectively. Streptococcus viridans was the leading microorganism isolated (21/50, 42%). Staphylococci and enterococci were found in 9 (18%) and 5 (10%) patients, respectively. Twelve patients (24%) were culture-negative in this series. Embolic complications occurred in 13 patients( 26%), with a total of 17 episodes. No significant correlation was found between the occurrence of embolization and the vegetation size or the location of the vegetation, if patients with right-sided valvular vegetation and no identifiable vegetation were excluded. Surgery was needed by 25 patients (50%), and mortality occurred in 6 (12%). TEE was superior to transthoracic echocardiography in the detection of vegetations at the mitral or prosthetic valves. Concerning the associated complications with infective endocarditis, TEE was also superior in estimating the severity of mitral regurgitation, recognizing ruptured chordae tendineae and detecting subaortic complications such as valve ring abscess and mitral valve perforation.( ABSTRACT TRUNCATED AT 250 WORDS)#A149313

Clinical Use of Clopidogrel in Acute Coronary Syndrome

Several therapeutic approaches have been developed to improve the outcome among patients with acute coronary syndrome (ACS). However, treatment with antithrombotic therapies such as oral glycoprotein IIb/IIIa inhibitors has been limited by the lack of efficacy and excess bleeding complications. As the publication of the landmark study Clopidogrel in Unstable Angina to Prevent Recurrent Events ( CURE), the clinical benefit of early and intermediate-term use of combined antiplatelet agents - clopidogrel plus aspirin - in non-ST-segment elevation myocardial infarction( NSTEMI) patients became evident. Pretreatment and intermediate -term therapy with clopidogrel in NSTEMI ACS patients undergoing percutaneous coronary intervention (PCI) was further supported by the PCI- CURE trial. Recently, the results of two major trials Clopidogrel as Adjunctive Reperfusion Therapy-Thrombolysis in Myocardial Infarction 28 , Clopidogrel and Metoprolol in Myocardial Infarction Trial established the pivotal role of clopidogrel in the other spectrum of ACS-STEMI. Coupled with the results from previous multicentre trials, these two studies provide a guide for the early and long-term use of clopidogrel in the whole spectrum of ACS. A review summarising the results of the recent clinical trials and a discussion on its implications for the clinical management of ACS is presented