大鼠急性心肌缺血早期标记物及其差异基因筛选的研究
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摘要
第一部分SELDI-TOF MS筛选大鼠急性心肌缺血早期标记物的研究
背景冠心病在我国的发病率逐年上升，病死率和致残率居高不下，也是构成法医学猝死的主要原因。冠心病病人约有1／4无明显的胸痛等症状，约50％无心电图改变。法医学实践中也发现：如果心肌缺血发生在室间隔等重要部位，即使是微小的心肌梗死，也会引起心源性猝死。临床上常用肌钙蛋白等作冠心病的辅助诊断，但该蛋白是确诊心肌梗死的指标，在血浆中要2-4 h以后才出现，更早的心肌缺血指标如C反应蛋白，心肌脂肪酸结合蛋白等特异性较差，需要和肌钙蛋白联合应用，检测方法繁复。因此，寻找新的特异性好的心肌缺血的早期标志物显得尤为重要。
SELDI-TOF MS是近年来新的蛋白质组分析工具，其原理是：用不同性质的化学芯片结合不同性质的蛋白质和多肽，经过能量分子结合后受激光的激发进入质谱，以其在质谱中的不同飞行时间来分离蛋白。该方法广泛用于寻找疾病蛋白标记。我们采用了SELDI-TOF MS的观察大鼠急性心肌缺血后的血清蛋白变化规律，寻找能作为心肌缺血的蛋白标记。同时我们也观察了心肌本身蛋白质谱的改变。
方法SD大鼠分成手术组，假手术组和非手术对照组。在结扎大鼠冠脉前降支，在5 min-6 h内不同的时间段取大鼠的静脉血和心肌组织。血清和心肌组织的蛋白提取物分别经过化学芯片的初步筛选后均选定CM10芯片作为实验用芯片。用Ciphergen公司的PBSⅡ-C型SELDI-TOF MS质谱仪检测。同时血清进行了cTnI和CKMB的检测，心肌进行常规HE染色。
结果只出现于手术组而几乎不出现于假手术组和非手术对照组的峰为特异峰；出现于手术组和假手术组而在非手术组中不出现的峰称非特异峰。血清CM10芯片检测到3个心肌缺血特异性峰，质荷比(m／z)分别是7564 Da、7586 Da、9583 Da。其中7564 Da、7586 Da相伴出现，在缺血5min组阳性率50％，在15 min以后各缺血组的阳性率达到97％，在假手术组仅有1个样本有这两个峰，未见于非手术组，特异性达到97％。9583 Da仅存在于缺血后1 h以内。发现价在手术组和假手术组都出现的非心肌特异性峰(4983 Da、5140 Da、8075 Da、9423 Da)。4个峰值改变的非特异性峰：3个升高的峰(4400 Da、4542 Da、8400 Da)，1个在假手术组和手术组都降低的峰(7666 Da)。
缺血心肌组织同两个对照组比较，主要有两种差别峰：特异性出现的新峰和峰值升高的峰。特异性出现的新峰3个：质荷比(m／z)分别为6304Da、8337 Da、8376 Da，这三个峰在手术组中阳性率为100％，假手术组和非手术组中均没有发现，其中峰6304 Da，8337 Da在手术5 min后就升高，8376 Da也是在缺血5 min后出现，15 min后峰值明显升高。特异性升高的峰3个：其中质荷比(m／z)为6658 Da、6876 Da两个峰都是在实验组中15 min以后升高2倍以上，8577 Da峰则在缺血5 min以后升高。cTnI和CKMB在血清中的含量分别均在4 h以后升高。HE染色在1 h可见心肌收缩带，4-6 h可见有炎症细胞浸润。
结论血清中出现7564 Da、7586 Da蛋白峰，缺血心肌组织中出现6304Da、8337 Da、8376 Da蛋白峰都是急性心肌缺血后的特异性改变，是目前已知检测出的最早的蛋白标记物。有望用于临床心肌缺血的早期诊断和法医学心源性猝死的诊断。
第二部分大鼠急性心肌缺血后差异基因的筛选
背景了解心肌缺血后早期分子变化的机制，对于寻找疾病的标记和干预治疗缺血性心肌病意义重大。目前关于心肌缺血在转录水平上的研究多采用cDNA芯片的方法，该方法用已知的EST和基因做探针，其结果取决于芯片上探针的密度，而且对于低表达的RNA效果差，更无法了解可能出现的新基因。抑制消减杂交(SSH)是一种从转录水平比较两种材料mRNA差异的方法，其设计方法从一定程度上弥补了DNA芯片的不足。我们用该方法检测了大鼠心肌缺血后的差异表达基因。
方法8只SD大鼠心肌缺血1 h后处死，提取纯化mRNA，以缺血心肌作为Tester，非缺血自身对照作为Driver，进行正向杂交，以非缺血心肌作为Tester，缺血心肌作为Driver进行反向杂交。扩增产物构建消减cDNA文库，经过测序和斑点杂交的确认后，得到不同基因的克隆片断，并对测序结果进行生物学分析。
结果正向杂交得到的已知基因有16个：分为以下五类：(1)心肌和细胞骨架：myoglobin，troponin T2，cardiac(Tnnt2)，tropomyosin 1 alpha(Tpm1)，Rattus norvegicus beta-glo(MGC72973)。(2)蛋白翻译相关：ribosomal protein L39，ribosomal protein S11(Rps11)，eukaryotic translation elongation factor 1 alpha 1(Eef1a1)。(3)代谢以及相关酶类：ATPase 3(Psmc3)，malate dehydrogenase 1 NAD，lactate dehydrogenase B(Ldhb)，UDP-glucose pyrophosphorylase 2(Ugp2)。(4)心肌重塑相关：cathepsin L(Ctsl)。(5)其他：Sjogren syndrome antigen B(Ssb)，basigin(Bsg)，unc-45 homolog A(Unc45a)。反向杂交的到的已知基因有1个氧化还原相关蛋白NADH dehydrogenase(ubiquinone)Fe-S protein 2(Ndufs2)。共获得新克隆16个，其中9个可以在REFRNA库中查到并可根据NCBI的DNA数据库预测其功能：7个核糖体相关，一个为Telethonin，一个为actin。另外7个完全未知，无法根据DNA数据库预测其可能序列和功能。其中Sjogren syndrome antigen B(Ssb)，basigin(Bsg)，unc-45 homolog A(Unc45a)，cathepsin L(Ctsl)，Telethonin均为第一次发现与早期心肌缺血有关。其功能尚需进一步证实。
结论我们构建了大鼠心肌缺血后的消减杂交文库。五个基因第一次报道与心肌缺血有关，分别是Sjogren syndrome antigen B(Ssb)，basigin(Bsg)，unc-45 homolog A(Unc45a)，cathepsin L(Ctsl)和Telethonin，发现了7个新的EST与心肌缺血相关的序列。
Chapter 1 Potential biomarkers discorvery of acute myocardial ischemia using SELDI-TOF MS
Background The increasing prevalence of Coronary Artery Disease (CAD) has high mortality and morbidity. Besides, CAD constructs the major death cause of sudden death in forensic science. About 1/4 CAD patients have not pectoral syndrome and half of them do not show electrocardiogram changes. Forensic pathologist found that when the important area is ischemia, such as ischemia of ventricular septal, could lead to sudden cardiac death (SCD). Troponin (cTn) is widely used as ancillary clinical diagnosis of CAD. As a marker of myocardial infarction, cTn could be detected 2-4 h in the plasma after the onset of the clinical syndrome. The earlier biomarkers, such as CRP and HABP, had to be used for earlier diagnose of cardiac ischemia for their poor specificity. SELDI-TOF MS was novel proteomic tool. According to its principle, the proteins were captured by ProteinChips which can bind with different proteins or peptides for their different chemistry properties, after that, the proteins or peptides were bound with energy molecules and run in the mass spectrometry when inspired by laser. The peptides were separated by their flying time in the MS. This equipment was widely used to find disease biomarkers. In this study, SELDI TOF MS were used to detect the pepitides profile and biomarkers of cardiac ischemia in the sera and myocardium.
Methods Sprague-Dawley rats were divided as three groups: operation group, sham operation group, non-operation group. The sera and ischemia myocardium were collected 5 min-6 h after the coronary artery ligation. CM10 ProteinChip was selected as the experiment chip. The sera and myocardium protein extracts were processed by CM10 ProteinChip and analyzed by SELDI-TOF MS. CKMB, cTnI and histological change were detected too.
Result The peaks detected in the operation groups and nearly seen in the sham operation groups were called as specific peaks. Those peaks emerged in the operation groups and sham operation groups but not in the non-operation groups were called as non-specific groups. Three peaks with m/z 7564Da, 7586 Da and 9583 Da were only found in the operation groups' sera. The peaks 7564Da and 7586 Da were seen simultaneously had the sensitivity of 97.0% and specificity of 97.0% in the>15 min operation groups. Some non-specific peaks in response to injury have also been seen, they are the peaks with the m/z 4983 Da, 5140 Da, 8075 Da, 9423 Da. Four non-specifc peaks with marked peak height changes: 4400 Da, 4542 Da, 8400 Da(increased), and 7666 Da(decreased). Two kinds of peaks were found in the myocardium protein extraction. One is the three novel specific peaks which were only found in operation groups with the 100% specificity: 6304Da, 8337Da, 8376Da. Peak 6304Da, 8337Da were found with a high peak height >5 min operation group, while peak height of 8376Da was low in 5 min group but increased in the >15min groups. Another kind of peaks was specific peaks with increased peak height. Peak 6658 Da, 6876 Da increased in>15min groups, peak 8577 Da increased in >5 min groups. While the significant increase of CKMB and cTnI in the serum could be detected until 4 hours after the ligation. For the HE staining of the myocardium, contraction band was observed after 1 h of the ligation, inflammatory cells were found in the ischemia area 4-6 h after the ligation.
Conclusion Whether in the serum or the myocardium, the change peaks found by SELDI-TOF MS were the earliest changes after cardiac ischemia by now. These changes may reflect the modified peptides or novel proteion. They may be used as the biomarker of cardiac ischemia for clinical purpose or ancillary ischemia sigh for SCD.
Chaper 2 Screening expreesed genes in a rat model during cardiac ischemia by suppression subtractive hybridization
Background Understanding the molecular changes after the cardiac ischemia is pivotal for the diagnosis of myocardial infarction and therapy of the ischemia cardiac diseases. By now, The main approch used for research on myocardial ischemia transcription is microarray detection. This method can only detect the known genes and ESTs and limited by the molecule transcription level. Suppression Subtractive Hybridization (SSH) is a approch to detect the different transcription gene between two groups. The SSH is complementary to cDNA microaray. We detected the transcription changes after cardiac ischemia by SSH in this study.
Method Eight Sprague-Dawley rats were sacrificed after the coronary artery ligation for 1 h and the myocardium were collected. After the mRNA was extracted, a PCR-Select cDNA Substraction Kit was used as the manufacture's protocol. Two hybridizations were established: the forward hybridization: ischemia myocardium cDNA were used as the tester, and non-ischemia cDNA as the driver; the reverse hybridization: non-ischemia myocardium cDNA as the tester, and ischemia cDNA as the driver. The subtractive cDNA library were sequenced and identified by dot blot.
Result Sixteen known genes were sequenced after forward hybridization: they can be classified as the following: (1) Myocardium gene: myoglobin, cardiac troponin T2, tropomyosin 1, Rattus norvegicus beta-glo. (2)Translation related gene: ribosomal protein L39, ribosomal protein S11 (Rps11),eukaryotic translation elongation factor 1 alpha 1. (3) Metabolism and enzymes: ATPase 3 (Psmc3), malate dehydrogenase 1, NAD, lactate dehydrogenase B (Ldhb), UDP-glucose pyrophosphorylase 2 (Ugp2).(4) Myocardium remodeling: cathepsin L (Cts1). (5) Others: Sjogren syndrome antigen B (Ssb), basigin (Bsg), unc-45 homolog A (Unc45a). Only one known gene, NADH dehydrogenase (ubiquinone) Fe-S protein 2, was found in the reverse hybridization. Sixteen unknown clones were detected: Nine of them can be aligned in the REFRNA databank, including 7 sequences which were predicted similar to ribosomal protein, and two others were telethonin and actin; Seven of them were totally unknown about there function. Among them, Ssb, basigin, unc-45 homolog A (Unc45a),cathepsin L (Cts1) and Telethonin were first reported to be associated with acute myocardial ischemia.
Conclusion We established the SSH library of rat ischemia (ischemia duration for 1 h). Five genes including Ssb, basigin, unc-45 homolog A (Unc45a), cathepsin L (Cts1) and Telethonin were first found to be associated with acute myocardial ischemia. Seven totally new ESTs were found.
背景冠心病在我国的发病率逐年上升，病死率和致残率居高不下，也是构成法医学猝死的主要原因。冠心病病人约有1／4无明显的胸痛等症状，约50％无心电图改变。法医学实践中也发现：如果心肌缺血发生在室间隔等重要部位，即使是微小的心肌梗死，也会引起心源性猝死。临床上常用肌钙蛋白等作冠心病的辅助诊断，但该蛋白是确诊心肌梗死的指标，在血浆中要2-4 h以后才出现，更早的心肌缺血指标如C反应蛋白，心肌脂肪酸结合蛋白等特异性较差，需要和肌钙蛋白联合应用，检测方法繁复。因此，寻找新的特异性好的心肌缺血的早期标志物显得尤为重要。
SELDI-TOF MS是近年来新的蛋白质组分析工具，其原理是：用不同性质的化学芯片结合不同性质的蛋白质和多肽，经过能量分子结合后受激光的激发进入质谱，以其在质谱中的不同飞行时间来分离蛋白。该方法广泛用于寻找疾病蛋白标记。我们采用了SELDI-TOF MS的观察大鼠急性心肌缺血后的血清蛋白变化规律，寻找能作为心肌缺血的蛋白标记。同时我们也观察了心肌本身蛋白质谱的改变。
方法SD大鼠分成手术组，假手术组和非手术对照组。在结扎大鼠冠脉前降支，在5 min-6 h内不同的时间段取大鼠的静脉血和心肌组织。血清和心肌组织的蛋白提取物分别经过化学芯片的初步筛选后均选定CM10芯片作为实验用芯片。用Ciphergen公司的PBSⅡ-C型SELDI-TOF MS质谱仪检测。同时血清进行了cTnI和CKMB的检测，心肌进行常规HE染色。
结果只出现于手术组而几乎不出现于假手术组和非手术对照组的峰为特异峰；出现于手术组和假手术组而在非手术组中不出现的峰称非特异峰。血清CM10芯片检测到3个心肌缺血特异性峰，质荷比(m／z)分别是7564 Da、7586 Da、9583 Da。其中7564 Da、7586 Da相伴出现，在缺血5min组阳性率50％，在15 min以后各缺血组的阳性率达到97％，在假手术组仅有1个样本有这两个峰，未见于非手术组，特异性达到97％。9583 Da仅存在于缺血后1 h以内。发现价在手术组和假手术组都出现的非心肌特异性峰(4983 Da、5140 Da、8075 Da、9423 Da)。4个峰值改变的非特异性峰：3个升高的峰(4400 Da、4542 Da、8400 Da)，1个在假手术组和手术组都降低的峰(7666 Da)。
缺血心肌组织同两个对照组比较，主要有两种差别峰：特异性出现的新峰和峰值升高的峰。特异性出现的新峰3个：质荷比(m／z)分别为6304Da、8337 Da、8376 Da，这三个峰在手术组中阳性率为100％，假手术组和非手术组中均没有发现，其中峰6304 Da，8337 Da在手术5 min后就升高，8376 Da也是在缺血5 min后出现，15 min后峰值明显升高。特异性升高的峰3个：其中质荷比(m／z)为6658 Da、6876 Da两个峰都是在实验组中15 min以后升高2倍以上，8577 Da峰则在缺血5 min以后升高。cTnI和CKMB在血清中的含量分别均在4 h以后升高。HE染色在1 h可见心肌收缩带，4-6 h可见有炎症细胞浸润。
结论血清中出现7564 Da、7586 Da蛋白峰，缺血心肌组织中出现6304Da、8337 Da、8376 Da蛋白峰都是急性心肌缺血后的特异性改变，是目前已知检测出的最早的蛋白标记物。有望用于临床心肌缺血的早期诊断和法医学心源性猝死的诊断。
第二部分大鼠急性心肌缺血后差异基因的筛选
背景了解心肌缺血后早期分子变化的机制，对于寻找疾病的标记和干预治疗缺血性心肌病意义重大。目前关于心肌缺血在转录水平上的研究多采用cDNA芯片的方法，该方法用已知的EST和基因做探针，其结果取决于芯片上探针的密度，而且对于低表达的RNA效果差，更无法了解可能出现的新基因。抑制消减杂交(SSH)是一种从转录水平比较两种材料mRNA差异的方法，其设计方法从一定程度上弥补了DNA芯片的不足。我们用该方法检测了大鼠心肌缺血后的差异表达基因。
方法8只SD大鼠心肌缺血1 h后处死，提取纯化mRNA，以缺血心肌作为Tester，非缺血自身对照作为Driver，进行正向杂交，以非缺血心肌作为Tester，缺血心肌作为Driver进行反向杂交。扩增产物构建消减cDNA文库，经过测序和斑点杂交的确认后，得到不同基因的克隆片断，并对测序结果进行生物学分析。
结果正向杂交得到的已知基因有16个：分为以下五类：(1)心肌和细胞骨架：myoglobin，troponin T2，cardiac(Tnnt2)，tropomyosin 1 alpha(Tpm1)，Rattus norvegicus beta-glo(MGC72973)。(2)蛋白翻译相关：ribosomal protein L39，ribosomal protein S11(Rps11)，eukaryotic translation elongation factor 1 alpha 1(Eef1a1)。(3)代谢以及相关酶类：ATPase 3(Psmc3)，malate dehydrogenase 1 NAD，lactate dehydrogenase B(Ldhb)，UDP-glucose pyrophosphorylase 2(Ugp2)。(4)心肌重塑相关：cathepsin L(Ctsl)。(5)其他：Sjogren syndrome antigen B(Ssb)，basigin(Bsg)，unc-45 homolog A(Unc45a)。反向杂交的到的已知基因有1个氧化还原相关蛋白NADH dehydrogenase(ubiquinone)Fe-S protein 2(Ndufs2)。共获得新克隆16个，其中9个可以在REFRNA库中查到并可根据NCBI的DNA数据库预测其功能：7个核糖体相关，一个为Telethonin，一个为actin。另外7个完全未知，无法根据DNA数据库预测其可能序列和功能。其中Sjogren syndrome antigen B(Ssb)，basigin(Bsg)，unc-45 homolog A(Unc45a)，cathepsin L(Ctsl)，Telethonin均为第一次发现与早期心肌缺血有关。其功能尚需进一步证实。
结论我们构建了大鼠心肌缺血后的消减杂交文库。五个基因第一次报道与心肌缺血有关，分别是Sjogren syndrome antigen B(Ssb)，basigin(Bsg)，unc-45 homolog A(Unc45a)，cathepsin L(Ctsl)和Telethonin，发现了7个新的EST与心肌缺血相关的序列。
Chapter 1 Potential biomarkers discorvery of acute myocardial ischemia using SELDI-TOF MS
Background The increasing prevalence of Coronary Artery Disease (CAD) has high mortality and morbidity. Besides, CAD constructs the major death cause of sudden death in forensic science. About 1/4 CAD patients have not pectoral syndrome and half of them do not show electrocardiogram changes. Forensic pathologist found that when the important area is ischemia, such as ischemia of ventricular septal, could lead to sudden cardiac death (SCD). Troponin (cTn) is widely used as ancillary clinical diagnosis of CAD. As a marker of myocardial infarction, cTn could be detected 2-4 h in the plasma after the onset of the clinical syndrome. The earlier biomarkers, such as CRP and HABP, had to be used for earlier diagnose of cardiac ischemia for their poor specificity. SELDI-TOF MS was novel proteomic tool. According to its principle, the proteins were captured by ProteinChips which can bind with different proteins or peptides for their different chemistry properties, after that, the proteins or peptides were bound with energy molecules and run in the mass spectrometry when inspired by laser. The peptides were separated by their flying time in the MS. This equipment was widely used to find disease biomarkers. In this study, SELDI TOF MS were used to detect the pepitides profile and biomarkers of cardiac ischemia in the sera and myocardium.
Methods Sprague-Dawley rats were divided as three groups: operation group, sham operation group, non-operation group. The sera and ischemia myocardium were collected 5 min-6 h after the coronary artery ligation. CM10 ProteinChip was selected as the experiment chip. The sera and myocardium protein extracts were processed by CM10 ProteinChip and analyzed by SELDI-TOF MS. CKMB, cTnI and histological change were detected too.
Result The peaks detected in the operation groups and nearly seen in the sham operation groups were called as specific peaks. Those peaks emerged in the operation groups and sham operation groups but not in the non-operation groups were called as non-specific groups. Three peaks with m/z 7564Da, 7586 Da and 9583 Da were only found in the operation groups' sera. The peaks 7564Da and 7586 Da were seen simultaneously had the sensitivity of 97.0% and specificity of 97.0% in the>15 min operation groups. Some non-specific peaks in response to injury have also been seen, they are the peaks with the m/z 4983 Da, 5140 Da, 8075 Da, 9423 Da. Four non-specifc peaks with marked peak height changes: 4400 Da, 4542 Da, 8400 Da(increased), and 7666 Da(decreased). Two kinds of peaks were found in the myocardium protein extraction. One is the three novel specific peaks which were only found in operation groups with the 100% specificity: 6304Da, 8337Da, 8376Da. Peak 6304Da, 8337Da were found with a high peak height >5 min operation group, while peak height of 8376Da was low in 5 min group but increased in the >15min groups. Another kind of peaks was specific peaks with increased peak height. Peak 6658 Da, 6876 Da increased in>15min groups, peak 8577 Da increased in >5 min groups. While the significant increase of CKMB and cTnI in the serum could be detected until 4 hours after the ligation. For the HE staining of the myocardium, contraction band was observed after 1 h of the ligation, inflammatory cells were found in the ischemia area 4-6 h after the ligation.
Conclusion Whether in the serum or the myocardium, the change peaks found by SELDI-TOF MS were the earliest changes after cardiac ischemia by now. These changes may reflect the modified peptides or novel proteion. They may be used as the biomarker of cardiac ischemia for clinical purpose or ancillary ischemia sigh for SCD.
Chaper 2 Screening expreesed genes in a rat model during cardiac ischemia by suppression subtractive hybridization
Background Understanding the molecular changes after the cardiac ischemia is pivotal for the diagnosis of myocardial infarction and therapy of the ischemia cardiac diseases. By now, The main approch used for research on myocardial ischemia transcription is microarray detection. This method can only detect the known genes and ESTs and limited by the molecule transcription level. Suppression Subtractive Hybridization (SSH) is a approch to detect the different transcription gene between two groups. The SSH is complementary to cDNA microaray. We detected the transcription changes after cardiac ischemia by SSH in this study.
Method Eight Sprague-Dawley rats were sacrificed after the coronary artery ligation for 1 h and the myocardium were collected. After the mRNA was extracted, a PCR-Select cDNA Substraction Kit was used as the manufacture's protocol. Two hybridizations were established: the forward hybridization: ischemia myocardium cDNA were used as the tester, and non-ischemia cDNA as the driver; the reverse hybridization: non-ischemia myocardium cDNA as the tester, and ischemia cDNA as the driver. The subtractive cDNA library were sequenced and identified by dot blot.
Result Sixteen known genes were sequenced after forward hybridization: they can be classified as the following: (1) Myocardium gene: myoglobin, cardiac troponin T2, tropomyosin 1, Rattus norvegicus beta-glo. (2)Translation related gene: ribosomal protein L39, ribosomal protein S11 (Rps11),eukaryotic translation elongation factor 1 alpha 1. (3) Metabolism and enzymes: ATPase 3 (Psmc3), malate dehydrogenase 1, NAD, lactate dehydrogenase B (Ldhb), UDP-glucose pyrophosphorylase 2 (Ugp2).(4) Myocardium remodeling: cathepsin L (Cts1). (5) Others: Sjogren syndrome antigen B (Ssb), basigin (Bsg), unc-45 homolog A (Unc45a). Only one known gene, NADH dehydrogenase (ubiquinone) Fe-S protein 2, was found in the reverse hybridization. Sixteen unknown clones were detected: Nine of them can be aligned in the REFRNA databank, including 7 sequences which were predicted similar to ribosomal protein, and two others were telethonin and actin; Seven of them were totally unknown about there function. Among them, Ssb, basigin, unc-45 homolog A (Unc45a),cathepsin L (Cts1) and Telethonin were first reported to be associated with acute myocardial ischemia.
Conclusion We established the SSH library of rat ischemia (ischemia duration for 1 h). Five genes including Ssb, basigin, unc-45 homolog A (Unc45a), cathepsin L (Cts1) and Telethonin were first found to be associated with acute myocardial ischemia. Seven totally new ESTs were found.
引文
1 Lopez AD, Murray CCJL. The global burden of disease, 1990-2020. Nature Med 1998;4:1241-1243
2 Yellon DM, Baxter GF Protecting the ischaemic and reperfused myocardium in acute myocardial infarction: distant dream or near reality? Heart 2000;83:381-387
3 Alpert JS, Thygesen K, Antman E, et al. Myocardial infarction redefined-a consensus document of The Joint European Society of Cardiology/American College of Cardiology Committee for the redefinition of myocardial infarction. J Am Coll Cardiol 2000;36:959-969
4 朱伟，赵子彦，靳刚．急性心肌梗死诊断的生物化学标志物研究进展．国外医学临床生物化学与检验学分册．2005：26：86-88
5 Cohle SD, Sampson BA. The negative autopsy: Sudden cardiac death or other? Cardiovas Pathol 2001;10:271-274
6 司雅芳，姚仲义，司彦明，等．96例心源性猝死分析．中国法医学杂志 2003：18(5)：282-283
7 江一清，刘朝中，朱国英．现代冠心病学．北京：人民军医出版社 2001：879
8 徐英含．实用法医病理学．群众出版社 1984：352-365
9 Berhane BT, Zong C, Liem DA, etal. Cardiovascular-related proteins identified in human plasma by the HUPO Plasma Proteome Project Pillot Phase. Proteomics 2005;5:3520-3530
10 Prentice H, Webster KA. Genomic and proteomic profilesof heart disease. Trends Cardiovasc Med 2004;14:282-288
11 Hutchens TW, Yip TT. Rapid Commun. Mass Spectrom. 1993;7:576-580
12 Merchant M, Weinberger SR. Recent advancement in surface enhancement laser desorption/ionization of flight mass spectrometry. Electrophoresis 2000;21:1164-1167
13 Pasinetti GM, Ungar LH, Lange DJ, et al. Identification of potential CSF biomarkers in ALS. Clin Chem 2006;52:1501-1509
14 Rieg S, Steffen H, Seeber S, et al. Deficiency of dermcidin-derived antimicrobial peptides in sweat of patients with atopic dermatitis correlates with an impaired innate defense of human skin in vivo. J Immunol 2005;174:8003-8010
15 Khurana M, Traum AZ, Aivado M, et al. Urine proteomic profiling of pediatric nephrotic syndrome. Pediatr Nephrol 2006;21:1257-1265
16 Oh JH, Gao J, Nandi A, et al. Diagnosis of early relapse in ovarian cancer using serum proteomic profiling. Genome Inform 2005;16:195-204
17 成官迅，张雪林，刘伊丽．急性心肌梗塞后危险区与梗塞区的识别．中国医学影像技术，2002：18：1098-1099
18 Adam BL, Qu Y, Davis JW, et al. Serum protein fingerprinting coupled with a pattern-matching algorithm distinguishes prostate cancer from benign prostate hyperplasia and healthy men. Cancer Res 2002;62:3609-3614
19 VlahouA, Schellhamer PF, Mendrinos S, et al. Development of a novel proteomic approach for the detection of transitional cell carcinoma of the bladder in urine. Am J Pathol 2001;158:1491-1502
20 Gion M, Mione R, Leon AE, et al. Comparison of the diagnostic accuracy of CA27-29 and CA15-3 in primary breast cancer. Clin Chem 1999;45:630-637
21 Watkins B, Szaro R, Ball S, et al. Detection of early-stage cancer by serum protein analysis. Am Lab 2001;33:32-36
22 Mateos-Caceres PJ, Garcia-Mendez A, Lopez Farre A, et al. Proteomic Analysis of Plasma From Patients During an Acute Coronary Syndrome. J Am Coll Cardiol 2004;44:1578-1581
23 Melanson SF, Tanasijevic MJ. Laboratory diagnosis of acute myocardial injury. Cardiovas Pathol 2005;14:156-161
24 魏晓真 利用飞行质谱法对急性心肌梗死血清标志物的研究．中国急救医学 2004：24：715-717
25 Schipper R, Loof A, de Groot J, etal. SELDI-TOF-MS of saliva: methodology and pre-treatment effects. J Chromatogr B 2007;847:45-53
1 Diatchenko L,Lau Y,Campbell AP,etal. Suppression substractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. PNAS 1996; 93: 6025-6030
2 Jj W,wright MB,Cai L,et al. Efficacy of SSH PCR in isolating differentially expressed genes. BMC Genomics 2002; 20: 12-16
3 Lethe B,Iacas S,Miehaux L,et al. LAGE-1,a new gene with tumor specificity. Int J cancer 1998; 76: 903-908
4 Yuan BZ,Miller MJ,Keck CL,et al. Cloning,characterization,and chromosome localization of a gene frequently deleted in human liver cancerr(D1C-I)homologous to rat RhogAP. Cancer Res1998; 58: 2196-2199
5 Zhou J,Liu ZH,Wang XQ,et al. Combining SSH and reverse northem for esophageal carcinoma of diferentially expressed genes. Chinese Science Bulletin 2001; 46: 222-225.
6 Wang JH,Shan YJ. Identification of differentially expressed genes of acute hypoxia-treated HepG2 cells and hypoxia acclimatized HepG2 cells. Acta Physiologica Sinica 2003; 53: 324-330.
7 肖忠海，洪欣，尹昭云．抑制消减杂交技术研究进展及其在环境医学上的应用．环境与健康杂志 2005；22：497-499
8 王振英，彭永康，郑坚瑜．黑麦中与耐盐性相关的cDNA片段的克隆与序列分析．植物研究 2005；25：427-430
9 Chim SS,Cheung SSF,Tsui SKW. Differential gene expression of rat neonatal heart analyzed by suppression subtractive hybridization and expressed sequence tag sequencing. J Cell Biochem 2000; 80: 24-36
10 周炜，赵寿元，李本昌等．抑制PCR技术及其在基因分析中的应用．高技术通讯．1999．55-59
11 Sakai J,Ishikawa H,Kojima S,Proteomic analysis of rat heart in ischemia and ischemia-reperfusion using fluorescence two-dimensional difference gel electrophoresis. Proteomics 2003; 3: 318-1324
12 White MY,Cordwell SJ,McCarron HCK,et al. Proteomics of ischemia/reperfusion injury in rabbit myocardium reveals alterations to proteins of essential functional systems. Proteomics 2005; 5: 1395-1410
13 Stanton LW,Garrard LJ,Damm D,et al.: 2000. Altered patterns of gene expression inresponse to myocardial infarction. Circ Res 86: 939-945
14 Harpster MH,Bandyopadhyay S,Thomas DP,et al. Earliest changes in the left ventricular transcriptome post-myocardial infarction. Mammalian Genome 2006; 17: 701-715
15 White MY,Cordwell SJ,McCarron HCK,et al. Proteomics of ischemia/reperfusion injury in rabbit myocardium reveals alterations to proteins of essential functional systems. Proteomics 2005; 5: 1395-1410
16 Simkhovich BZ,Marjoram P,Poizatd C,et al. Brief episode of ischemia activates protective genetic program in rat heart: a gene chip study. Cardiovasc Res 2003; 59: 450-459
17 Kim SJ,Yatner DE,Topper JN,et al. Gene program for cardiac cell survival induced by transient ischemia in conscious pigs. PNAS 2001; 98: 9336-9341
18 骞爱荣，商澎，陈志南．CD147结构与功能研究进展．中国免疫学杂志 2003；19：728-731
19 Hori K,Katayama N,Kachi S,et al. Retinal Dysfunction in Basigin Deficiency. Invest Ophthalmol Vis Sci 2000; 41: 3128-3133
20 Han M,Trotta P,Coleman C,et al. MCT-4,A511/Basigin and EF5 expression patterns during early chick cardiomyogenesis indicate cardiac cell differentiation occurs in a hypoxic environment. Dev Dyn 2006; 235: 124-131
21 Burns-Kurtis CL,Olzinski AR,Needle S,et al. Cathepsin S expression is upregulated following balloon angiop lasty in the hypercholesterolemic rabbit. Cardiovasc Res 2004; 62: 610-620.
22 刘玉胜，鹿庆华，蒋卫东．组织蛋白酶与细胞外基质重塑．基础医学与临床 2006：26(9)：1037-1040
23 Stoka V,Turk B,Schendel SL,et al. Lysosomal p rotease pathways to apoptosisl Cleavage of bid,not p rocaspases,is the most likely route. J Biol Chem 2001; 276: 3149-3157.
24 于小华，张新刚，李双杰．组织蛋白酶L在扩张型心肌病心肌中的表达及其意义．中华内科杂志 2005；44：495-498
25 Aispaugh M,Maddison P. Resolution of the identity of certain antigen antibody system in systemic lupus erthematosus and Sjogren's syndrome: an interlaboratory collaboration. Arthritis Rheum 1979; 22: 796-798.
26 Zeng X,Miura T,Luo Y,et al. Properties of pluripotent human embryonic stem cells BG01 and BG02. Stem Cells 2004; 22: 292-312
27 Zhang H,Zhou L,Yang R,et al. Identification of differentially expressed genes in human heart with ventricular septal defect using suppression subtractive hybridization. Biochem Biophys Res Commu 2006:342:135-144
28 Hutagalung A H, Landsverk MG, Price MG, et al. The UCS family of myosin chaperones. J Cell Sci 2002;115:3983-3990
29 Valle G, Faulkner G, De Antoni A, et al. Telethonin, a novel sarcomeric p rotein of heart and skeletal muscle. FEBS Lett 1997;415:163-168
30 Moreira ES, Wiltshire TJ, Faulkner G, et al. Limb-girdle muscular dystrophy type 2G is caused by mutations in the gene encoding the sarcomeric p rotein telethonin . Nat Genet 2000;24:163-166
31 Nicholas G, Thomas M, Langley B, et al. Titin-Cap associates with, and regulates secretion of, Myostatin. J Cell Physiol 2002;193:120-131
32 Granzier HL, Labeit S. The Giant Protein Titin: A Major Player in Myocardial Mechanics, Signaling and Disease. Circ Res 2004;94:284-295
1 Lawler W. The negative coroner's necropsy: a personal approach and consideration of difficulties. J Clin Pathol 1990; 43: 977-980
2 Cohle SD,Sampson BA. The negative autopsy: Sudden cardiac death or other? Cardiovascu Pathol 2001; 2: 219-222
3 Priori SG,Aliot E,Blomstrom-Lundquist,et al. task force on sudden cardiac death of the European Society of Cardiology. Eur Heart J 2001; 22: 1374-450
4 Myerburg RJ,Castellanos A. Cardiac arrest and sudden cardiac death. In: Braunwald E,editor. Heart disease: a textbook of cardiovascular medicine. New York: WB Saunders; 1997; 756-79
5 Thygesena K,Uretsky BF. Acute ischaemia as a trigger of sudden cardiac death. Eur Heart J Supplements 2004(Supplement D): D88-D90
6 江一清，刘朝中，朱国英．现代冠心病学．北京：人民军医出版社 2001：879
7 Nishida N,Ikeda N,Katayama Y,et al. Subendocardial small infarct in the superior ventricular septum as a cause of sudden death. Forensic Sci Int 2003; 138: 62-67
8 Lecomte D,Fornesb P,Nicolas G. Stressful events as a trigger of sudden death: a study of 43 medico-legal autopsy cases. Forensic Sci Int 1996; 79: 1-10
9 Coronado BE ,Griffith JL,Beshanaky JR,et al. Hospital mortality in women and men with acute cardiac ischemia: A prospective multicenter study. JACC 1997; 29(7): 1490-1496
10 徐成斌，主编．临床心血管病学．第1版．北京：北京科学技术出版社 1997：229
11 Tunstall-Pedoe H,Kuulasmaa K,Amouyel P,et al. Myocardial infarction and coronary deaths in the World Health Organization MONICA Project. Circulation 1994; 90: 583
12 吕卓人，唐海波，急性心肌梗死的早期诊断中国实验诊断学 2003：7：12—13
13 Alpert JS,Thygesen K,Antman E,et al. Myocardial infarction redefined-a consensus document of The Joint European Society of Cardiology/American College of Cardiology Committee for the redefinition of myocardial infarction. J Am Coll Cardiol 2000; 36: 959-969
14 Zhu BL,Ishikawa T,Michiue T. Postmortem cardiac troponin T levels in the blood and pericardial fluid. Part 2: Analysis for application in the diagnosis of sudden cardiac death with regard to pathology. Legal Med 2006; 8: 94-101
15 徐英含．实用法医病理学．群众出版社 1984：352-365
16 刘英绅，钟志玖，班翔，等．诊断早期心肌缺血的三种组织学染色方法的比较．哈尔滨医科大学学报，2002；36：238-240
17 Zabel M,Hohnloser SH,Koster W,et al. Analysis of creatine kinase ,CK-MB,myoglobin,and troponin T time-activity curves for early assessment of coronary artery reperfusion after intravenous thromolysis. Circulation 1993; 87: 1542-1548
18 Puleo PR,Meyer D,Wathen C,et al. Use of a rapid assay of subforms of creatine kinase MB to diagnose or rule out acute myocardial infarction. N Engl J Med 1994; 331: 561-569
19 Jaffe AS,Babuin L,Apple FS. Biomarkers in Acute Cardiac Disease: the Present and the Future. J Am Coll Cardiol 2006; 48: 1-11
20 Melanson SF, Tanasijevic MJ. Laboratory diagnosis of acute myocardial injury. Cardiovascu Pathol 2005:14:156-161
21 Devaraj S, DuClos TW, Jialal I. Binding and internalization of Oreactive protein by Fc-gamma receptors on human aortic endothelial cells mediates biological effects. Arterioscler Thromb Vasc Biol 2005;25:1359 - 63
22 Ridker PM, Rifai N, Rose L, et al. Comparison of Oreactive protein and low-density lipoprotein cholesterol levels in the prediction of first cardiovascular events. N Engl J Med 2002;347:1557-1565
23 Danesh J, Wheeler JG, Hirschfield GM, et al. C-reactive protein and other circulating markers of inflammation in the prediction of coronary heart disease. N Engl J Med 2004;350:1387-1397
24 de Lemos JA, Morrow DA, Bentley JH, et al. The prognostic value of B-type natriuretic peptide in patients with acute coronary syndromes. N Engl J Med 2001:345:1014-1021
25 Jernberg T, StridsbergM, Venge P, et al. N-terminal pro brain natriuretic peptide on admission for early risk stratification of patients with chest pain and no ST-segment elevation. J Am Coll Cardiol 2002;40:437-445
26 Sinha MK, Roy D, Gaze DC, et al. Role of ischemia modified albumin, a new biochemical marker of myocardial ischaemia, in the early diagnosis of acute coronary syndromes. Emerg Med J 2004;21:29-34
27 Chan CP, Sanderson JE, Glatz JF, et al. A superior early myocardial infarction marker Human heart-type fatty acid-binding protein. Z Kardiol 2004:93:388-397
28 Pelsers MM, Hermens WT, Glatz JF. Fatty acid-binding proteins as plasma markers of tissue injury. Clin Chem Acta 2005:352:15-35
29 Heeschen C,Dimmeler S,Hamm CW,et al. Soluble CD40 ligand in acute coronary syndromes. N Engl J Med 2003; 348: 1104-1111
30 Brennan ML,Penn MS,Van Lente F,et al. Prognostic value of myeloperoxidase in patients with chest pain. N Engl J Med 2003; 349: 1595-1604
31 Bayes-Genis A,Conover CA,Overgaard MT,et al. Pregnancy associated plasma protein A as a marker of acute coronary syndromes. N Engl J Med 2001; 345: 1022-1029
32 Higo S,Uemat su M ,Yamagishi M ,et al. Elevation of plasma matrix metalloproteinase-9 in the culprit coronary artery in patients with acute myocardial infarction: clinical evidence from distal protection. Circ J 2005; 69: 1180-1185
33 Guray U,Erbay AR,Guray Y,et al. Levels of soluble adhesion molecules in various clinical presentations of coronary at herosclerosis. Int J Cardiol 2004; 96: 235-240
34 Heeschen C,Dimmeler S,Fichtlscherer S,et al. Prognostic value of placental growth factor in patients with acute chest pain. JAMA 2004; 291: 435-41
35 Apple FS,Wu AHB,Mair J,et al. Future Biomarkers for Detection of Ischemia and Risk Stratification in Acute Coronary Syndrome. Clinical Chemistry 2005; 51: 810-824
36 Fallon JT. Simplified method for histochemical demonstration of experimental myocardial infarct. Circulation 1979; 60: 11-42
37 方云祥，汤显良，陈修等．家兔实验性心肌梗塞模型的改进和梗塞范围定量的动态观察．湖南医学院学报 1980；5：229-232
38 江宏，刘世沧，董培正，等，实验性大鼠急性心肌缺血的组织化学和扫描电镜观察．中国法医学杂志 1994；9：200—222
39 邵同先，张苏亚，赵立法．苏木素碱性品红苦味酸染色对显示家兔急性心肌缺血的观察．河南医学研究 2003；12：124-126
40 刘英绅，钟志玖，班翔，等．诊断早期心肌缺血的三种组织学染色方法的比较．哈尔滨医科大学学报，2002；36：238-240
41 张永亮，李灵芝，孙慧宽等．急性心肌缺血的早期病理学变化及诊断方法．中国法医学杂志 1993；8：190-193
42 Fine G,Morales A,Scerpella J. R,et al. Experimental myocardial infarction: A histochemical study. Arch Pathol 1986; 82: 4-10
43 Thomsen H,Held H. Immunohistochemical detection of C5b-9(m) in myocardium: an aid in distintuishing infarction induced ischemic heart muscle necrosis from other forms of lethal myocardial injury. Forensic Sci Int 1995; 71: 87-93
44 Kent SP. Intracellular plasma protein: a manifestation of cell injury in myocardial ischeamia. Nature 1966; 21: 1279-1281
45 Shekhonin BV,Gurivev SB,Irgashev SB,et al. Immunofluorecent identification of fibronectin and fibrinogen/fibrin in experimental myocardial infarction. J Mol Cell Cardiol 1990; 22: 533-541
46 胡丙杰，冯绮明，汪冠三，等．心肌梗死6项免疫组化指标的死后稳定性比较．中国法医学杂志 2003；4：208-221
47 Thornell L E ,Holmbom B ,griksson A ,et al. Enzyme and immunohistochemical assessment of myocardial damage after ischaemia and reperfusion in a closed2chest pig model. Histochemistry 1992; 98: 341-345
48 申成斌，王兆福，胡丙杰．实验性早期心肌梗死的免疫组化研究．河南医学研 究 2003；12：119-123
49 胡丙杰，陈玉川，祝家镇，等．纤维连接蛋白诊断心肌梗死的死后稳定性研究。中国法医学杂志 2000；15：7-9
50 Hansen SH,Rossen K,Evaluation of cardiac troponin Ⅰ immunoreaction in autopsy hearts: a possible marker of early myocardial infarction. Forensic Sci Int 1999; 99: 189-196
51 Block MI,Myocardial myoglobin following coronary artery occulation. Am J Path 1983; 111: 374-378
52 Nomoto KI,Mori N,Shoji T,et al. Early loss of myocardial myoglobin detected immunohistochemically following occlusion of the coronary artery in rats. Exp Mol Pathol 1987; 47: 390-402
53 姚季生，姚申，应爱娣．试验性早期心肌梗死的肌红蛋白免役组织化学研究．法医学杂志 1992；8：49-51
54 Zhang JM,Riddick LR. Cytoskeleton immunohistochemical study of early ischemic myocardium. Forensic Sci Int 1996; 80: 229-238
55 陈建国，徐小虎，姚青松，等．VEGF在早期心肌梗死死后诊断中的应用．中国法医学杂志，2000；15：14-17
56 王志勇，李政，陈新山．大鼠急性心肌缺血早期内皮素的免疫组化染色观察．中国法医学杂志 2003；18：205-207
57 赵江民，倪炯，李铭．心肌梗死早期病理改变的实验研究同济大学学报(医学版)2005；26：12-15
58 陈建国，徐小虎．急性心肌缺血早期碱性纤维细胞生长因子的免疫组化研究．法医学杂志 1998：14：71-73
59 Loennechen JP,Stoylen A,Beisvag V,et al. Regional expression of endothelin-1,ANP,IGF-1,and LV wall stress in the infarcted rat heart. Am J Physiol Heart Circ Physiol 2001; 280,H2902-H2910
60 Zugibe FT,Jr B P,Conley T,et al. Determination of myocardial alterations at autopsy in the absence of gross and microscopic changes. Arch Path 1966; 81: 409-413
61 张耀痒，邵同先，赵立法等．实验性急性心肌梗死心肌K+／Na+，Mg2+／Ca2+，Zn+／Cu+比值变化的研究．中国法医学杂志 1993；8：143-144
62 Cina SJ,Li DJ,Chan DW,et al. Serum concentrations of cardiac troponin Ⅰ in sudden death: a pilot study. Am J Forensic Med Pathol,1998; 19: 324-328
63 张永亮，李灵芝，王久亮，等．心肌氨基酸代谢与早期急性心肌缺血的关系．中国法医学杂志 1993；8：83-85
64 Pereira L,Richards M,Goios A,et al. Evaluating the forensic informativeness of mtDNA haplogroup H sub-typing on a Eurasian scale. Forensic Sci Int 2006; 159: 43-50
65 胡丙杰，陈玉川，祝家镇，等．实验性早期心肌缺血c-Fos原位杂交研究．中国法医学杂志 2000；15：133-135
66 Bukowska A,Lendeckel U,Kahne T,et al. ProteOmics in myocardial diseases. Pathol Res Pract 2004; 200: 135-45
67 Mizukami Y,Iwamatsu A ,Aki r,et al. ERK1/2 Regulates Intracellular ATP Levels through α-Enolase Expression in Cardiomyocytes Exposed to Ischemic Hypoxia and Reoxygenation. J Biol Chem 2004,279(48): 50120-50131
68 Berhane BT,Zong C,Liem DA,etal. Cardiovascular-related proteins identified in human plasma by the HUPO Plasma Proteome Project Pilot Phase. Proteomics 2005; 5: 3520-3530
69 White MY,Cordwell SJ,McCarron HCK,Proteomics of ischemia/reperfusion injury in rabbit myocardium reveals alterations to proteins of essentialfunctional systems. Proteomics 2005; 5: 1395-1410
70 Yan L,Vatner DE,Kim SJ,et al. Autophagy in chronically ischemic myocardium. PNAS,2005; 102: 13807-13812
71 冯雪，叶能胜，张荣利，等．大鼠急性心肌梗死模型比较蛋白组学的初步研究．分析试验室 2006；25：23-27
72 Mateos-Caceres PJ,Garcia-Mendez A,Lopez Farre A,et al. Proteomic analysis of plasma from patients during an acute coronary syndrome. J Am Coll Cardiol 2004; 44: 1578-1581
73 Greijer AE,van der Groep P,Kemming D,etal. Up-regulation of gene expression by hypoxia is mediated predominantly by hypoxia-inducible factor 1 (HIF-1). J Pathol 2005; 206: 291-304
74 杜林林，王朝晖．低氧诱导因子-1α在大鼠心肌梗死中的表达及其意义．中国中西医结合急救杂志 2006；13(6)：367-369
75 魏晓真 利用飞行质谱法对急性心肌梗死血清标志物的研究．中国急救医学 2004；24(10)：715-717
76 高春芳，郑国宝，赵光，等．急性心肌梗死病程不同时期患者血清蛋白质组学分析．解放军医学杂志 2005；30：465-466
77 Winslow RL,Boguski MS,Genome Informatics: Current Status and Future Prospects. Circ Res 2003; 92: 953-961
78 Stanton L,Garrard L,DammD,et al. Altered patterns of gene expression in response to myocardial infartion. Circ Res 2000; 86: 939-945
79 Lyn D,Liu X,Nicole A,et al. Gene expression profile in mouse myocardium after ischemia. Physiol Genomics 2000; 2: 93-100
80 Harpster MH, Bandyopadhyay S, Thomas DP, et al.Earliest changes in the left ventricular transcriptome post-myocardial infarction. Mammalian Genome 2006;17:701-715
2 Yellon DM, Baxter GF Protecting the ischaemic and reperfused myocardium in acute myocardial infarction: distant dream or near reality? Heart 2000;83:381-387
3 Alpert JS, Thygesen K, Antman E, et al. Myocardial infarction redefined-a consensus document of The Joint European Society of Cardiology/American College of Cardiology Committee for the redefinition of myocardial infarction. J Am Coll Cardiol 2000;36:959-969
4 朱伟，赵子彦，靳刚．急性心肌梗死诊断的生物化学标志物研究进展．国外医学临床生物化学与检验学分册．2005：26：86-88
5 Cohle SD, Sampson BA. The negative autopsy: Sudden cardiac death or other? Cardiovas Pathol 2001;10:271-274
6 司雅芳，姚仲义，司彦明，等．96例心源性猝死分析．中国法医学杂志 2003：18(5)：282-283
7 江一清，刘朝中，朱国英．现代冠心病学．北京：人民军医出版社 2001：879
8 徐英含．实用法医病理学．群众出版社 1984：352-365
9 Berhane BT, Zong C, Liem DA, etal. Cardiovascular-related proteins identified in human plasma by the HUPO Plasma Proteome Project Pillot Phase. Proteomics 2005;5:3520-3530
10 Prentice H, Webster KA. Genomic and proteomic profilesof heart disease. Trends Cardiovasc Med 2004;14:282-288
11 Hutchens TW, Yip TT. Rapid Commun. Mass Spectrom. 1993;7:576-580
12 Merchant M, Weinberger SR. Recent advancement in surface enhancement laser desorption/ionization of flight mass spectrometry. Electrophoresis 2000;21:1164-1167
13 Pasinetti GM, Ungar LH, Lange DJ, et al. Identification of potential CSF biomarkers in ALS. Clin Chem 2006;52:1501-1509
14 Rieg S, Steffen H, Seeber S, et al. Deficiency of dermcidin-derived antimicrobial peptides in sweat of patients with atopic dermatitis correlates with an impaired innate defense of human skin in vivo. J Immunol 2005;174:8003-8010
15 Khurana M, Traum AZ, Aivado M, et al. Urine proteomic profiling of pediatric nephrotic syndrome. Pediatr Nephrol 2006;21:1257-1265
16 Oh JH, Gao J, Nandi A, et al. Diagnosis of early relapse in ovarian cancer using serum proteomic profiling. Genome Inform 2005;16:195-204
17 成官迅，张雪林，刘伊丽．急性心肌梗塞后危险区与梗塞区的识别．中国医学影像技术，2002：18：1098-1099
18 Adam BL, Qu Y, Davis JW, et al. Serum protein fingerprinting coupled with a pattern-matching algorithm distinguishes prostate cancer from benign prostate hyperplasia and healthy men. Cancer Res 2002;62:3609-3614
19 VlahouA, Schellhamer PF, Mendrinos S, et al. Development of a novel proteomic approach for the detection of transitional cell carcinoma of the bladder in urine. Am J Pathol 2001;158:1491-1502
20 Gion M, Mione R, Leon AE, et al. Comparison of the diagnostic accuracy of CA27-29 and CA15-3 in primary breast cancer. Clin Chem 1999;45:630-637
21 Watkins B, Szaro R, Ball S, et al. Detection of early-stage cancer by serum protein analysis. Am Lab 2001;33:32-36
22 Mateos-Caceres PJ, Garcia-Mendez A, Lopez Farre A, et al. Proteomic Analysis of Plasma From Patients During an Acute Coronary Syndrome. J Am Coll Cardiol 2004;44:1578-1581
23 Melanson SF, Tanasijevic MJ. Laboratory diagnosis of acute myocardial injury. Cardiovas Pathol 2005;14:156-161
24 魏晓真 利用飞行质谱法对急性心肌梗死血清标志物的研究．中国急救医学 2004：24：715-717
25 Schipper R, Loof A, de Groot J, etal. SELDI-TOF-MS of saliva: methodology and pre-treatment effects. J Chromatogr B 2007;847:45-53
1 Diatchenko L,Lau Y,Campbell AP,etal. Suppression substractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. PNAS 1996; 93: 6025-6030
2 Jj W,wright MB,Cai L,et al. Efficacy of SSH PCR in isolating differentially expressed genes. BMC Genomics 2002; 20: 12-16
3 Lethe B,Iacas S,Miehaux L,et al. LAGE-1,a new gene with tumor specificity. Int J cancer 1998; 76: 903-908
4 Yuan BZ,Miller MJ,Keck CL,et al. Cloning,characterization,and chromosome localization of a gene frequently deleted in human liver cancerr(D1C-I)homologous to rat RhogAP. Cancer Res1998; 58: 2196-2199
5 Zhou J,Liu ZH,Wang XQ,et al. Combining SSH and reverse northem for esophageal carcinoma of diferentially expressed genes. Chinese Science Bulletin 2001; 46: 222-225.
6 Wang JH,Shan YJ. Identification of differentially expressed genes of acute hypoxia-treated HepG2 cells and hypoxia acclimatized HepG2 cells. Acta Physiologica Sinica 2003; 53: 324-330.
7 肖忠海，洪欣，尹昭云．抑制消减杂交技术研究进展及其在环境医学上的应用．环境与健康杂志 2005；22：497-499
8 王振英，彭永康，郑坚瑜．黑麦中与耐盐性相关的cDNA片段的克隆与序列分析．植物研究 2005；25：427-430
9 Chim SS,Cheung SSF,Tsui SKW. Differential gene expression of rat neonatal heart analyzed by suppression subtractive hybridization and expressed sequence tag sequencing. J Cell Biochem 2000; 80: 24-36
10 周炜，赵寿元，李本昌等．抑制PCR技术及其在基因分析中的应用．高技术通讯．1999．55-59
11 Sakai J,Ishikawa H,Kojima S,Proteomic analysis of rat heart in ischemia and ischemia-reperfusion using fluorescence two-dimensional difference gel electrophoresis. Proteomics 2003; 3: 318-1324
12 White MY,Cordwell SJ,McCarron HCK,et al. Proteomics of ischemia/reperfusion injury in rabbit myocardium reveals alterations to proteins of essential functional systems. Proteomics 2005; 5: 1395-1410
13 Stanton LW,Garrard LJ,Damm D,et al.: 2000. Altered patterns of gene expression inresponse to myocardial infarction. Circ Res 86: 939-945
14 Harpster MH,Bandyopadhyay S,Thomas DP,et al. Earliest changes in the left ventricular transcriptome post-myocardial infarction. Mammalian Genome 2006; 17: 701-715
15 White MY,Cordwell SJ,McCarron HCK,et al. Proteomics of ischemia/reperfusion injury in rabbit myocardium reveals alterations to proteins of essential functional systems. Proteomics 2005; 5: 1395-1410
16 Simkhovich BZ,Marjoram P,Poizatd C,et al. Brief episode of ischemia activates protective genetic program in rat heart: a gene chip study. Cardiovasc Res 2003; 59: 450-459
17 Kim SJ,Yatner DE,Topper JN,et al. Gene program for cardiac cell survival induced by transient ischemia in conscious pigs. PNAS 2001; 98: 9336-9341
18 骞爱荣，商澎，陈志南．CD147结构与功能研究进展．中国免疫学杂志 2003；19：728-731
19 Hori K,Katayama N,Kachi S,et al. Retinal Dysfunction in Basigin Deficiency. Invest Ophthalmol Vis Sci 2000; 41: 3128-3133
20 Han M,Trotta P,Coleman C,et al. MCT-4,A511/Basigin and EF5 expression patterns during early chick cardiomyogenesis indicate cardiac cell differentiation occurs in a hypoxic environment. Dev Dyn 2006; 235: 124-131
21 Burns-Kurtis CL,Olzinski AR,Needle S,et al. Cathepsin S expression is upregulated following balloon angiop lasty in the hypercholesterolemic rabbit. Cardiovasc Res 2004; 62: 610-620.
22 刘玉胜，鹿庆华，蒋卫东．组织蛋白酶与细胞外基质重塑．基础医学与临床 2006：26(9)：1037-1040
23 Stoka V,Turk B,Schendel SL,et al. Lysosomal p rotease pathways to apoptosisl Cleavage of bid,not p rocaspases,is the most likely route. J Biol Chem 2001; 276: 3149-3157.
24 于小华，张新刚，李双杰．组织蛋白酶L在扩张型心肌病心肌中的表达及其意义．中华内科杂志 2005；44：495-498
25 Aispaugh M,Maddison P. Resolution of the identity of certain antigen antibody system in systemic lupus erthematosus and Sjogren's syndrome: an interlaboratory collaboration. Arthritis Rheum 1979; 22: 796-798.
26 Zeng X,Miura T,Luo Y,et al. Properties of pluripotent human embryonic stem cells BG01 and BG02. Stem Cells 2004; 22: 292-312
27 Zhang H,Zhou L,Yang R,et al. Identification of differentially expressed genes in human heart with ventricular septal defect using suppression subtractive hybridization. Biochem Biophys Res Commu 2006:342:135-144
28 Hutagalung A H, Landsverk MG, Price MG, et al. The UCS family of myosin chaperones. J Cell Sci 2002;115:3983-3990
29 Valle G, Faulkner G, De Antoni A, et al. Telethonin, a novel sarcomeric p rotein of heart and skeletal muscle. FEBS Lett 1997;415:163-168
30 Moreira ES, Wiltshire TJ, Faulkner G, et al. Limb-girdle muscular dystrophy type 2G is caused by mutations in the gene encoding the sarcomeric p rotein telethonin . Nat Genet 2000;24:163-166
31 Nicholas G, Thomas M, Langley B, et al. Titin-Cap associates with, and regulates secretion of, Myostatin. J Cell Physiol 2002;193:120-131
32 Granzier HL, Labeit S. The Giant Protein Titin: A Major Player in Myocardial Mechanics, Signaling and Disease. Circ Res 2004;94:284-295
1 Lawler W. The negative coroner's necropsy: a personal approach and consideration of difficulties. J Clin Pathol 1990; 43: 977-980
2 Cohle SD,Sampson BA. The negative autopsy: Sudden cardiac death or other? Cardiovascu Pathol 2001; 2: 219-222
3 Priori SG,Aliot E,Blomstrom-Lundquist,et al. task force on sudden cardiac death of the European Society of Cardiology. Eur Heart J 2001; 22: 1374-450
4 Myerburg RJ,Castellanos A. Cardiac arrest and sudden cardiac death. In: Braunwald E,editor. Heart disease: a textbook of cardiovascular medicine. New York: WB Saunders; 1997; 756-79
5 Thygesena K,Uretsky BF. Acute ischaemia as a trigger of sudden cardiac death. Eur Heart J Supplements 2004(Supplement D): D88-D90
6 江一清，刘朝中，朱国英．现代冠心病学．北京：人民军医出版社 2001：879
7 Nishida N,Ikeda N,Katayama Y,et al. Subendocardial small infarct in the superior ventricular septum as a cause of sudden death. Forensic Sci Int 2003; 138: 62-67
8 Lecomte D,Fornesb P,Nicolas G. Stressful events as a trigger of sudden death: a study of 43 medico-legal autopsy cases. Forensic Sci Int 1996; 79: 1-10
9 Coronado BE ,Griffith JL,Beshanaky JR,et al. Hospital mortality in women and men with acute cardiac ischemia: A prospective multicenter study. JACC 1997; 29(7): 1490-1496
10 徐成斌，主编．临床心血管病学．第1版．北京：北京科学技术出版社 1997：229
11 Tunstall-Pedoe H,Kuulasmaa K,Amouyel P,et al. Myocardial infarction and coronary deaths in the World Health Organization MONICA Project. Circulation 1994; 90: 583
12 吕卓人，唐海波，急性心肌梗死的早期诊断中国实验诊断学 2003：7：12—13
13 Alpert JS,Thygesen K,Antman E,et al. Myocardial infarction redefined-a consensus document of The Joint European Society of Cardiology/American College of Cardiology Committee for the redefinition of myocardial infarction. J Am Coll Cardiol 2000; 36: 959-969
14 Zhu BL,Ishikawa T,Michiue T. Postmortem cardiac troponin T levels in the blood and pericardial fluid. Part 2: Analysis for application in the diagnosis of sudden cardiac death with regard to pathology. Legal Med 2006; 8: 94-101
15 徐英含．实用法医病理学．群众出版社 1984：352-365
16 刘英绅，钟志玖，班翔，等．诊断早期心肌缺血的三种组织学染色方法的比较．哈尔滨医科大学学报，2002；36：238-240
17 Zabel M,Hohnloser SH,Koster W,et al. Analysis of creatine kinase ,CK-MB,myoglobin,and troponin T time-activity curves for early assessment of coronary artery reperfusion after intravenous thromolysis. Circulation 1993; 87: 1542-1548
18 Puleo PR,Meyer D,Wathen C,et al. Use of a rapid assay of subforms of creatine kinase MB to diagnose or rule out acute myocardial infarction. N Engl J Med 1994; 331: 561-569
19 Jaffe AS,Babuin L,Apple FS. Biomarkers in Acute Cardiac Disease: the Present and the Future. J Am Coll Cardiol 2006; 48: 1-11
20 Melanson SF, Tanasijevic MJ. Laboratory diagnosis of acute myocardial injury. Cardiovascu Pathol 2005:14:156-161
21 Devaraj S, DuClos TW, Jialal I. Binding and internalization of Oreactive protein by Fc-gamma receptors on human aortic endothelial cells mediates biological effects. Arterioscler Thromb Vasc Biol 2005;25:1359 - 63
22 Ridker PM, Rifai N, Rose L, et al. Comparison of Oreactive protein and low-density lipoprotein cholesterol levels in the prediction of first cardiovascular events. N Engl J Med 2002;347:1557-1565
23 Danesh J, Wheeler JG, Hirschfield GM, et al. C-reactive protein and other circulating markers of inflammation in the prediction of coronary heart disease. N Engl J Med 2004;350:1387-1397
24 de Lemos JA, Morrow DA, Bentley JH, et al. The prognostic value of B-type natriuretic peptide in patients with acute coronary syndromes. N Engl J Med 2001:345:1014-1021
25 Jernberg T, StridsbergM, Venge P, et al. N-terminal pro brain natriuretic peptide on admission for early risk stratification of patients with chest pain and no ST-segment elevation. J Am Coll Cardiol 2002;40:437-445
26 Sinha MK, Roy D, Gaze DC, et al. Role of ischemia modified albumin, a new biochemical marker of myocardial ischaemia, in the early diagnosis of acute coronary syndromes. Emerg Med J 2004;21:29-34
27 Chan CP, Sanderson JE, Glatz JF, et al. A superior early myocardial infarction marker Human heart-type fatty acid-binding protein. Z Kardiol 2004:93:388-397
28 Pelsers MM, Hermens WT, Glatz JF. Fatty acid-binding proteins as plasma markers of tissue injury. Clin Chem Acta 2005:352:15-35
29 Heeschen C,Dimmeler S,Hamm CW,et al. Soluble CD40 ligand in acute coronary syndromes. N Engl J Med 2003; 348: 1104-1111
30 Brennan ML,Penn MS,Van Lente F,et al. Prognostic value of myeloperoxidase in patients with chest pain. N Engl J Med 2003; 349: 1595-1604
31 Bayes-Genis A,Conover CA,Overgaard MT,et al. Pregnancy associated plasma protein A as a marker of acute coronary syndromes. N Engl J Med 2001; 345: 1022-1029
32 Higo S,Uemat su M ,Yamagishi M ,et al. Elevation of plasma matrix metalloproteinase-9 in the culprit coronary artery in patients with acute myocardial infarction: clinical evidence from distal protection. Circ J 2005; 69: 1180-1185
33 Guray U,Erbay AR,Guray Y,et al. Levels of soluble adhesion molecules in various clinical presentations of coronary at herosclerosis. Int J Cardiol 2004; 96: 235-240
34 Heeschen C,Dimmeler S,Fichtlscherer S,et al. Prognostic value of placental growth factor in patients with acute chest pain. JAMA 2004; 291: 435-41
35 Apple FS,Wu AHB,Mair J,et al. Future Biomarkers for Detection of Ischemia and Risk Stratification in Acute Coronary Syndrome. Clinical Chemistry 2005; 51: 810-824
36 Fallon JT. Simplified method for histochemical demonstration of experimental myocardial infarct. Circulation 1979; 60: 11-42
37 方云祥，汤显良，陈修等．家兔实验性心肌梗塞模型的改进和梗塞范围定量的动态观察．湖南医学院学报 1980；5：229-232
38 江宏，刘世沧，董培正，等，实验性大鼠急性心肌缺血的组织化学和扫描电镜观察．中国法医学杂志 1994；9：200—222
39 邵同先，张苏亚，赵立法．苏木素碱性品红苦味酸染色对显示家兔急性心肌缺血的观察．河南医学研究 2003；12：124-126
40 刘英绅，钟志玖，班翔，等．诊断早期心肌缺血的三种组织学染色方法的比较．哈尔滨医科大学学报，2002；36：238-240
41 张永亮，李灵芝，孙慧宽等．急性心肌缺血的早期病理学变化及诊断方法．中国法医学杂志 1993；8：190-193
42 Fine G,Morales A,Scerpella J. R,et al. Experimental myocardial infarction: A histochemical study. Arch Pathol 1986; 82: 4-10
43 Thomsen H,Held H. Immunohistochemical detection of C5b-9(m) in myocardium: an aid in distintuishing infarction induced ischemic heart muscle necrosis from other forms of lethal myocardial injury. Forensic Sci Int 1995; 71: 87-93
44 Kent SP. Intracellular plasma protein: a manifestation of cell injury in myocardial ischeamia. Nature 1966; 21: 1279-1281
45 Shekhonin BV,Gurivev SB,Irgashev SB,et al. Immunofluorecent identification of fibronectin and fibrinogen/fibrin in experimental myocardial infarction. J Mol Cell Cardiol 1990; 22: 533-541
46 胡丙杰，冯绮明，汪冠三，等．心肌梗死6项免疫组化指标的死后稳定性比较．中国法医学杂志 2003；4：208-221
47 Thornell L E ,Holmbom B ,griksson A ,et al. Enzyme and immunohistochemical assessment of myocardial damage after ischaemia and reperfusion in a closed2chest pig model. Histochemistry 1992; 98: 341-345
48 申成斌，王兆福，胡丙杰．实验性早期心肌梗死的免疫组化研究．河南医学研 究 2003；12：119-123
49 胡丙杰，陈玉川，祝家镇，等．纤维连接蛋白诊断心肌梗死的死后稳定性研究。中国法医学杂志 2000；15：7-9
50 Hansen SH,Rossen K,Evaluation of cardiac troponin Ⅰ immunoreaction in autopsy hearts: a possible marker of early myocardial infarction. Forensic Sci Int 1999; 99: 189-196
51 Block MI,Myocardial myoglobin following coronary artery occulation. Am J Path 1983; 111: 374-378
52 Nomoto KI,Mori N,Shoji T,et al. Early loss of myocardial myoglobin detected immunohistochemically following occlusion of the coronary artery in rats. Exp Mol Pathol 1987; 47: 390-402
53 姚季生，姚申，应爱娣．试验性早期心肌梗死的肌红蛋白免役组织化学研究．法医学杂志 1992；8：49-51
54 Zhang JM,Riddick LR. Cytoskeleton immunohistochemical study of early ischemic myocardium. Forensic Sci Int 1996; 80: 229-238
55 陈建国，徐小虎，姚青松，等．VEGF在早期心肌梗死死后诊断中的应用．中国法医学杂志，2000；15：14-17
56 王志勇，李政，陈新山．大鼠急性心肌缺血早期内皮素的免疫组化染色观察．中国法医学杂志 2003；18：205-207
57 赵江民，倪炯，李铭．心肌梗死早期病理改变的实验研究同济大学学报(医学版)2005；26：12-15
58 陈建国，徐小虎．急性心肌缺血早期碱性纤维细胞生长因子的免疫组化研究．法医学杂志 1998：14：71-73
59 Loennechen JP,Stoylen A,Beisvag V,et al. Regional expression of endothelin-1,ANP,IGF-1,and LV wall stress in the infarcted rat heart. Am J Physiol Heart Circ Physiol 2001; 280,H2902-H2910
60 Zugibe FT,Jr B P,Conley T,et al. Determination of myocardial alterations at autopsy in the absence of gross and microscopic changes. Arch Path 1966; 81: 409-413
61 张耀痒，邵同先，赵立法等．实验性急性心肌梗死心肌K+／Na+，Mg2+／Ca2+，Zn+／Cu+比值变化的研究．中国法医学杂志 1993；8：143-144
62 Cina SJ,Li DJ,Chan DW,et al. Serum concentrations of cardiac troponin Ⅰ in sudden death: a pilot study. Am J Forensic Med Pathol,1998; 19: 324-328
63 张永亮，李灵芝，王久亮，等．心肌氨基酸代谢与早期急性心肌缺血的关系．中国法医学杂志 1993；8：83-85
64 Pereira L,Richards M,Goios A,et al. Evaluating the forensic informativeness of mtDNA haplogroup H sub-typing on a Eurasian scale. Forensic Sci Int 2006; 159: 43-50
65 胡丙杰，陈玉川，祝家镇，等．实验性早期心肌缺血c-Fos原位杂交研究．中国法医学杂志 2000；15：133-135
66 Bukowska A,Lendeckel U,Kahne T,et al. ProteOmics in myocardial diseases. Pathol Res Pract 2004; 200: 135-45
67 Mizukami Y,Iwamatsu A ,Aki r,et al. ERK1/2 Regulates Intracellular ATP Levels through α-Enolase Expression in Cardiomyocytes Exposed to Ischemic Hypoxia and Reoxygenation. J Biol Chem 2004,279(48): 50120-50131
68 Berhane BT,Zong C,Liem DA,etal. Cardiovascular-related proteins identified in human plasma by the HUPO Plasma Proteome Project Pilot Phase. Proteomics 2005; 5: 3520-3530
69 White MY,Cordwell SJ,McCarron HCK,Proteomics of ischemia/reperfusion injury in rabbit myocardium reveals alterations to proteins of essentialfunctional systems. Proteomics 2005; 5: 1395-1410
70 Yan L,Vatner DE,Kim SJ,et al. Autophagy in chronically ischemic myocardium. PNAS,2005; 102: 13807-13812
71 冯雪，叶能胜，张荣利，等．大鼠急性心肌梗死模型比较蛋白组学的初步研究．分析试验室 2006；25：23-27
72 Mateos-Caceres PJ,Garcia-Mendez A,Lopez Farre A,et al. Proteomic analysis of plasma from patients during an acute coronary syndrome. J Am Coll Cardiol 2004; 44: 1578-1581
73 Greijer AE,van der Groep P,Kemming D,etal. Up-regulation of gene expression by hypoxia is mediated predominantly by hypoxia-inducible factor 1 (HIF-1). J Pathol 2005; 206: 291-304
74 杜林林，王朝晖．低氧诱导因子-1α在大鼠心肌梗死中的表达及其意义．中国中西医结合急救杂志 2006；13(6)：367-369
75 魏晓真 利用飞行质谱法对急性心肌梗死血清标志物的研究．中国急救医学 2004；24(10)：715-717
76 高春芳，郑国宝，赵光，等．急性心肌梗死病程不同时期患者血清蛋白质组学分析．解放军医学杂志 2005；30：465-466
77 Winslow RL,Boguski MS,Genome Informatics: Current Status and Future Prospects. Circ Res 2003; 92: 953-961
78 Stanton L,Garrard L,DammD,et al. Altered patterns of gene expression in response to myocardial infartion. Circ Res 2000; 86: 939-945
79 Lyn D,Liu X,Nicole A,et al. Gene expression profile in mouse myocardium after ischemia. Physiol Genomics 2000; 2: 93-100
80 Harpster MH, Bandyopadhyay S, Thomas DP, et al.Earliest changes in the left ventricular transcriptome post-myocardial infarction. Mammalian Genome 2006;17:701-715