冠心病患者血浆组织因子活性及凝血因子Ⅶ水平改变

目的研究汉族冠心病患者血浆中组织因子活性（aTF）、活化凝血因子Ⅶ（FⅦa）和凝血因子Ⅶ抗原（FⅦ：Ag）水平并探讨其与冠脉病变支数之间的关系。方法aTF采用发色底物法，FⅦa和FⅦ：Ag采用ELISA法。结果与对照组相比，冠心病患者血浆中aTF（P〈0．05）、FⅦa（P〈0．01）及FⅦ：Ag（P〈0．05）水平均显著升高；急性冠状动脉综合征（ACS）患者中aTF高于稳定型心绞痛（SAP）组（P〈0．05）和对照组（P〈0．01），后两者之间无显著差异；ACS和SAP患者之间血浆中FⅦa水平无显著差异，但均高于对照组；ACS组中FⅦ：Ag水平明显高于对照组。在不同冠脉病变支数的患者之间，aTF、FⅦa及FⅦ：Ag水平没有差异。结论SAP和ACS患者均可出现外源性凝血途径的激活，血浆中aTF增强可能预示了急性冠脉事件的发生；FⅦa可能可以作为冠心病的早期分子标志物；冠脉病变支数可能不能反映凝血激活的程度。     

冠心病患者血浆IL-10变化及其临床意义

目的探讨冠心病患者血浆IL-10变化及临床意义。方法选择急性冠脉综合征（ACS）患者（ACS组）25例,稳定型心绞痛（SAP）患者（SAP组）22例,正常对照组21例。采用ELISA法检测血浆IL-10、血糖及血脂。结果与对照组比较,ACS组、SAP组甘油三酯、低密度脂蛋白胆固醇、载脂蛋白B、脂蛋白a有统计学差异（P均〈0．05）。ACS组IL-10浓度明显高于SAP组和对照组（P均〈0．05）,而SAP组与对照组比较无统计学差异（P〉0．05）。IL-10与冠脉病变支数无相关性（r=-0．135,P〉0．05）。结论IL-10在ACS中明显升高,与冠脉病变支数无相关性。     

冠心病并发Ⅱ型糖尿病患者高凝状态的临床研究

目的探讨冠心病（CHD）并发非胰岛素依赖糖尿病（NIDDM）患者体内凝血功能的变化及临床意义。方法选择病例组CHD并发NIDDM患者24例,CHD患者32例,以及正常对照组20例,分别测定血浆D-二聚体（D-D im er）、纤维蛋白原（Fbg）含量,同时分析Fbg与冠脉病变支数的相关性。结果CHD并发NIDDM患者血浆D-D im er、Fbg水平显著高于CHD患者和正常对照者（P<0.05或P<0.01）,并且CHD患者也显著高于对照者（P<0.05）;冠脉病变严重者血浆Fbg含量显著高于冠脉病变较轻患者（P<0.01）。结论CHD并发NIDDM患者体内存在明显的高凝状态,并与冠状动脉病变程度有关。     

NT-proBNP、Hcy诊断急性冠脉综合征的临床研究

目的探讨NT—proBNP、Hcy诊断急性冠脉综合征（ACS）的临床意义。方法对70例AMI患者、50例UAP患者、50例SAP患者和50例健康体检者的血浆NT—proBNP、Hey水平进行比较分析。ACS患者均行冠状动脉造影术,并对其病变程度进行评估。结果AMI组和UAP组患者血浆NT—proBNP水平显著高于SAP组及对照组（P〈0．05）,而SAP组与对照组比较差异无统计学意义（P〉0．05）;ACS组、SAP组血浆Hcy水平均高于对照组,各组间比较差异有统计学意义（P〈O．05）。随着冠脉病变支数增加,ACS患者血浆NT—proBNP、Hcy水平逐渐升高（P〈0．05）。ACS患者中,心血管事件组血浆NT—proBNP、Hcy水平明显高于无心血管事件组（p〈0．05）。结论急性冠脉综合征患者的血浆NT—proBNP、Hcy水平均与冠状动脉粥样化病变程度密切相关,是ACS心血管不良事件的预测因子。     

血浆氧化低密度脂蛋白水平是冠心病严重程度的独立预报因素

目的研究血浆氧化低密度脂蛋白（OxLDL）水平与冠心病严重程度的关系。方法134例怀疑为冠心病的住院患者,进行选择性冠状动脉造影。根据冠脉造影结果,将患者分为冠心病组（113例,至少有1支冠脉狭窄≥50%）和对照组（21例,所有冠脉分支狭窄均<50%）。冠心病组按照冠脉病变支数进一步分为4个亚组（1支病变26例,2支病变26例,3支病变53例,4支病变8例）;按照发病症状分为稳定型心绞痛组（51例）、不稳定型心绞痛组（22例）和急性心肌梗死组（40例）。血浆OxLDL水平采用ELISA试剂盒检测。其他冠心病危险因素如年龄、性别、体质量指数、血脂等数据也一并收集。结果在冠心病组和对照组之间,除了高血压病史有显著升高外（46%vs19%,P<0.05）,其他指标没有显著差别。冠心病患者血浆OxLDL水平显著高于对照组（1.15±0.32）μkat/m lvs（0.68±0.30）μkat/m l,P<0.01)。14支病变冠心病患者,其血浆OxLDL含量均显著高于对照组（1.10±0.32）μkat/m l、（1.12±0.27）μkat/m l、（1.17±0.32）μkat/m l和（1.33±0.37）μkat/m lvs（0.68±0.30）μkat/m l,P<0.01);但不同支数病变患者亚组间无显著性差异。稳定型心绞痛、不稳定型心绞痛和急性心肌梗死患者血浆OxLDL含量均显著高于对照组（1.13±0.30）μkat/m l,（1.23±0.33）μkat/m l和（1.15±0.32）μkat/m lvs（0.68±0.30）μkat/m l,P<0.01);但是各分组之间无显著性差异。以血浆OxLDL水平为因变量进行多元回归分析,发现冠脉病变支数是血浆OxLDL水平升高的独立危险因素（P<0.01）。结论血浆OxLDL水平是冠心病严重程度独立预报因素。     

冠心病外周血白细胞PRKCB1 mRNA表达量与血管硬化危险因素及病变程度的相关性研究

目的 检测冠心病患者外周血白细胞蛋白激酶Cβ （PRKCB1）基因表达,分析其与冠心病类型、冠脉血管病变程度及动脉硬化危险因素血脂、高敏C-反应蛋白（hs-CRP）的关系.方法 纳入冠心病患者38例,其中稳定型心绞痛（SAP）10例、急性冠脉综合征（ACS）28例,另选健康对照组20名,进行血脂、hs-CRP水平、血细胞参数及PRKCB1 mRNA表达的检测分析.结果 SAP组和ACS组高密度脂蛋白胆固醇水平均较健康对照组显著降低（P＜0.05）,而甘油三酯水平高于健康对照组（P＜0.05, P＜0.01）,总胆固醇水平三组比较差异无统计学意义（P＞0.05）,SAP组和ACS组低密度脂蛋白胆固醇水平较健康对照组升高,ACS组升高差异有统计学意义（P＜0.05）.入院24 h后,与健康对照组相比,SAP组和ACS组患者hs-CRP水平升高,ACS组升高差异有统计学意义（P＜0.05）.SAP组和ACS组中性粒细胞计数均较健康对照组不同程度升高,ACS组升高差异有统计学意义（P＜0.05）.单核细胞计数和单核细胞百分比SAP组和ACS组均较健康对照组有升高趋势（P＞0.05）.冠心病患者PRKCB1 mRNA表达水平与健康对照者相比差异有统计学意义（P＜0.05）,且ACS组低于SAP组;PRKCB1 mRNA表达和血管病变支数呈负相关,单支、双支和三支血管病变组依次降低, 以三支血管病变组降低明显（P＜0.01）.结论 PRKCB1 mRNA表达与动脉粥样硬化危险因素明显相关,因冠心病的不同类型而有所差异,且与冠脉血管病变程度呈负相关.     

冠心病患者血浆IL-6及内脂素水平与冠状动脉病变严重程度相关性研究

目的 分析冠心病患者血浆白介素（IL）-6和内脂素水平与冠状动脉病变严重程度的相关性。方法 选择经冠状动脉造影确诊的冠心病患者128例,分为急性冠脉综合征（ACS）组和稳定型心绞痛（SAP）组,并按病变血管支数分成3组（单支病变组、双支病变组和三支病变组）,按冠状动脉狭窄程度分为3组（冠状动脉狭窄程度50%~75%组、76%~90%组及91%~100%组）。收集患者的基本临床资料,血浆IL-6及内脂素应用酶联免疫吸附法测定,应用Gensini积分评价冠状动脉病变严重程度。结果 ACS组患者IL-6及内脂素水平显著高于SAP组（P<0.05）,按照不同病变血管支数分组和按照病变血管狭窄程度分组后,血浆IL-6及内脂素水平在各组间差异具有显著性（P<0.05）。相关分析显示IL-6、内脂素与Gensini积分呈正相关（分别r=0.583,r=0.724,均P<0.01）。结论 IL-6及内脂素与冠状动脉病变严重程度密切相关。     

巨噬细胞移动抑制因子与冠状动脉病变的关系

目的探讨冠心病患者血浆巨噬细胞移动抑制因子（MIF）水平与冠状动脉粥样硬化病变的关系。方法根据选择性冠状动脉造影结果将142例患者分为冠心病组和对照组。冠心病组根据临床诊断分为稳定型心绞痛（SAP）和急性冠脉综合征（ACS）组;根据冠状动脉病变范围将冠心病组分为单支病变、双支病变和三支病变组。血浆MIF浓度通过ELISA法测定。结果冠心病组MIF水平明显高于对照组〔（14.97±4.11）vs（9.07±1.28）μg/L〕,ACS组MIF浓度又明显高于SAP组〔（16.66±3.56）vs（11.01±2.12）μg/L〕。随冠状动脉病变范围的增加,MIF浓度逐渐升高,各组间差异有显著统计学意义。结论MIF增高与冠状动脉粥样硬化发生显著相关,其可以做为预测冠状动脉粥样硬化病变情况的一个指标,与冠状动脉粥样硬化斑块不稳定关系更密切。     

血浆白三烯-B4浓度与冠状动脉斑块相关性的临床研究

目的通过测定不同类型冠心病患者血浆白三烯-B4(LT-B4)的水平,探讨其与冠心病(CHD)及冠状动脉病变的关系。方法入选400例患者,冠心病组310例,其中急性冠状动脉综合征(ACS)组217例,稳定性心绞痛(SAP)组93例,对照组90例。检测患者外周血浆LT-B4浓度。同时经选择性冠状动脉造影检查,明确冠状动脉病变情况。CHD组中85例患者接受64层螺旋CT冠状动脉成像检查,对冠状动脉内的主要斑块进行评价。结果冠心病组LT-B4水平明显高于对照组(t=12.365,P<0.05),ACS组LT-B4水平又明显高于SAP组(t=7.664,P<0.05)。随冠状动脉病变类型和冠状动脉病变程度的加重,LT-B4浓度逐渐升高;CHD患者的软斑块组、纤维斑块组较钙化斑块组血浆LT-B4浓度明显升高,差异有统计学意义;逐步回归分析表明LT-B4浓度与冠状动脉病变Gensini评分有显著相关性。结论 ACS患者白三烯-B4水平增高,并与冠脉病变的严重性明显正相关,血浆白三烯-B4检测结合64层螺旋CT测定能否提高判断ACS患者冠状动脉斑块性质的准确性,对ACS的诊断及对冠脉病变的判断有一定的预测作用。     

急性冠脉综合征患者血清纤维蛋白原、炎症因子检测的临床意义

目的 探讨冠心病患者凝血功能紊乱、炎症因子表达与动脉粥样硬化斑块不稳定性的关系,为冠心病发病机制的研究提供新的思路.方法 选择2007年10月至2008年8月于心内科行冠脉造影或冠脉CT患者195例,入院前均未行调脂治疗,按冠脉造影或冠脉CT结果分为健康对照组45例,稳定型心绞痛（SAP）组41例,急性冠脉综合征（ACS）组109例.分别于入院当时检测血清脂蛋白、纤维蛋白原（FIB）、单核细胞趋化蛋白-1（MCP-1）、高敏C反应蛋白（hs-CBP）水平.结果 冠心病患者的FIB、血清炎症因子MCP-1、hs-CRP、血脂水平明显升高（P〈0.05）,差异有统计学意义,表明患者存在炎症反应、脂代谢和凝血功能紊乱.与SAP组患者相比,ACS患者hs-CRP、FIB水平升高,差异有统计学意义,hs-CBP水平与FIB正相关,表明二者主要与动脉粥样硬化斑块的不稳定性相关.MCP-1与冠脉积分正相关,表明血清MCP-1水平可能反应冠脉病变的严重程度.结论 ACS患者血清hs-CRP、FIB较稳定型冠心病患者明显升高,二者呈相关关系,表明hs-CBP、FIB水平与冠脉粥样斑块的不稳定性相关,炎症反应和凝血异常可能相互促进,共同诱发斑块破裂,引起ACS.MCP-1与冠脉积分正相关,可能反映冠脉病变的严重程度.对ACS患者联合进行上述指标的检测有利于判断病情,尽早治疗,以改善预后.     

Standardization of factor VII/activated factor VII measurement in plasma of patients treated with recombinant factor activated VII.

To improve the standardization of the factor VII clotting activity (FVII:C) assay in patients treated with recombinant activated factor VII (rFVIIa), we conducted a multicentre study on plasma samples from four patients with haemophilia A, before and at various times after injection of a single dose of rFVIIa. FVII:C and prothrombin time were measured with the methods and reagents routinely used in each laboratory. Strong inter-laboratory variability of FVII:C values was found. The main source of variability was the type of thromboplastin. FVII:C values measured using rabbit thromboplastin were very close to activated factor VII clotting activity values (FVIIa:C) measured with a commercial assay (Staclot VIIa-TF). FVII:C values obtained with human placental thromboplastin were about three times lower than those obtained with rabbit and recombinant thromboplastins, and with the FVIIa:C assay. There was a good relationship between FVIIa:C and activated factor VII antigen values measured using a commercial immunoassay (Imubind FVIIa ELISA). In conclusion, rFVIIa at pharmacological concentrations can be easily monitored on the basis of FVII:C, using rabbit and probably also recombinant thromboplastin; equivalent results are obtained with a specific activated factor VII bioassay.     

Comparison of coagulant activity of factor VII and activated factor VII activity assays when used for determination of recombinant activated factor VII levels in plasma.

Two assays [coagulant activity of factor VII (FVII:C) and activated factor VII (FVIIa) activity] are currently available for the assessment of factor VII and FVIIa pharmacokinetics. This article presents the results of a comparison of the two assays when applied both in vitro as well as during clinical pharmacokinetic trials of recombinant FVIIa (rFVIIa) administered to healthy individuals and haemophilia patients. The in-vitro data showed that, for the FVII:C assay, plasma samples do not dilute in parallel. For the FVIIa activity assay, dilutions of samples are both parallel and linear with different dilutions of the calibrator. Moreover, intra-assay variation was found to be smaller for the FVIIa activity assay than for the FVII:C assay. When adding different amounts of rFVIIa (0-6 microg/ml) to normal plasma, a mean specific activity of rFVIIa of 48.6 U/mug was observed on applying the FVII:C assay; however, the specific activity decreased with increasing levels of rFVIIa. For the FVIIa activity assay, the mean specific activity was 45.4 IU/mug. Direct comparison of the two activity assays showed that no simple conversion between FVII:C and FVIIa activity measurements are possible. When applying the two assays for pharmacokinetic assessments in two clinical trials, statistically significant different estimates for the area under the curve, half-life, clearance and volume of distribution were obtained. In conclusion, for evaluation of rFVIIa pharmacokinetic properties, activity should be measured with the FVIIa activity assay - which is a more specific and reliable assay of the two available factor VII activity assays, especially when assessing low activity levels.     

Quantitation of activated factor VII levels in plasma using a tissue factor mutant selectively deficient in promoting factor VII activation

Although the majority of factor VII (FVII) circulates in the zymogen form, low levels of activated factor VII (FVIIa) have been postulated to exist in plasma and to serve a priming function for triggering of the clotting cascade. However, direct measurement of plasma FVIIa has not previously been possible. We have quantified plasma FVIIa levels using a novel, highly sensitive assay that is free from interference by FVII. Specificity of this clot-based assay results from the use of a mutant tissue factor that is selectively deficient in promoting FVII activation, but retains FVIIa cofactor function. In normal adults, FVIIa was found to be present in plasma (mean: 3.6 ng/mL) with considerable variation between individuals (range: 0.5 to 8.4 ng/mL). FVIIa levels were only loosely correlated with FVII coagulant activity, but were elevated in pregnancy and reduced with oral anticoagulant therapy. Incubation of plasma on ice in glass containers (cold activation) resulted in substantial FVIIa generation. Measurement of plasma forms of factor VII is of potential clinical importance because elevated FVII coagulant activity has been implicated as a significant risk predictor for ischemic heart disease. Clinically, this new assay will now permit direct assessment of the role of plasma FVIIa in thrombotic disorders.     

In vitro effects of heparin and tissue factor pathway inhibitor on factor VII assays. possible implications for measurements in vivo after heparin therapy.

The coagulant activity of blood coagulation factor VII (FVII:C) can be lowered by changes in lifestyle and by therapeutic intervention, e.g. heparin infusion. The question is, however, whether FVII:C determined ex vivo is a valid measure of the FVII activity in vivo. We measured plasma FVII:C, activated FVII (FVIIa), FVII protein (FVII:Ag), tissue factor pathway inhibitor (TFPI), triglycerides, and free fatty acids (FFA) before and 15 min after infusion of a bolus of unfractionated heparin (50 IU/kg body weight) in 12 healthy subjects. Additionally, we conducted in vitro experiments to investigate the effect of unfractionated heparin and TFPI, which is released from the endothelium by heparin, on FVII:C, FVIIa, and FVII:Ag. Heparin infusion decreased triglycerides and increased FFA and TFPI. This was accompanied by significant reductions in FVIIa, FVII:C and FVII:Ag. In vitro, anti-TFPI antibodies increased FVIIa and FVII:C, and heparin reduced FVIIa. The heparinase Hepzyme was unable to abolish the effect of heparin. There were no in vitro effects on FVII:Ag. We conclude that, due to interference by TFPI and heparin in post-heparin plasma, it is impossible to measure the in vivo FVII activity by means of FVII clotting assays. These assays should therefore not be used to measure the coagulation status of patients in heparin therapy, unless extraordinary precautions are taken to eliminate TFPI and heparin effects ex vivo. The observed effect of heparin on FVII:Ag should be investigated further.     

Elevation of factor VII activity and mass in coronary artery disease of varying severity.

The present study was undertaken to determine whether the extent of Factor VII elevation correlated with the severity of coronary artery disease and whether zymogen or activated Factor VII was responsible for this elevation. A group of 69 patients with coronary artery disease with old myocardial infarction was compared with 28 control subjects. The patient groups showed elevated levels of Factor VII procoagulant activity (FVII:C) and more markedly elevated Factor VII antigen (FVII:Ag) levels than the control group; therefore they had a decreased FVII:C to FVII:Ag ratio. The increased Factor VII level in the patient groups was caused by elevated Factor VII zymogen levels, and not by activated Factor VII. Since FVII:C levels strongly correlated with the titer of thrombin-antithrombin III complexes in all patients, the hypercoagulable state accompanying severe coronary atherosclerosis seems to underlie the increase of FVII and TAT in the stable phase of myocardial infarction.     

Dietary factor VII activation does not increase plasma concentrations of prothrombin fragment 1+2 in patients with stable angina pectoris and coronary atherosclerosis

Studies in healthy subjects showed that blood coagulation factor VII (FVII) is activated postprandially after consumption of high-fat meals, but accompanying thrombin formation has not been demonstrated. In patients with coronary atherosclerosis, the arterial intima is supposed to present more tissue factor, the cofactor of FVII, to circulating blood; therefore, thrombin formation in response to FVII activation is more likely to occur in such patients. This hypothesis was tested in a randomized crossover study of 30 patients (aged 43 to 70 years) with stable angina pectoris and angiographically verified coronary atherosclerosis. They were served a low-fat (5% of energy from fat) breakfast and lunch and a high-fat (40% of energy from fat) breakfast and lunch on 2 different days. Venous blood samples were collected at 8:15 AM (fasting), 12:30 PM, 2:00 PM, 3:30 PM, and 4:45 PM and analyzed for triglycerides, activated FVII (FVIIa), FVII protein concentration (FVII:Ag), prothrombin fragment 1+2 (F1+2), and soluble fibrin. Triglyceride levels increased from fasting levels on both diets, but they increased most markedly on the high-fat diet. FVIIa and FVIIa/FVII:Ag increased with the high-fat diet and decreased with the low-fat diet. For both diets, FVII:Ag and F1+2 decreased slightly. No postprandial changes were observed for soluble fibrin. Postprandial mean values of triglycerides, FVIIa, FVII:Ag, and FVIIa/FVII:Ag were significantly higher for the high-fat diet than for the low-fat diet. Our findings confirm that high-fat meals cause immediate activation of FVII. The clinical implication is debatable because FVII activation was not accompanied by an increase in plasma F1+2 concentrations in patients with severe atherosclerosis. However, a local thrombin generation on the plaque surface cannot be excluded.     

Heparin lowers plasma levels of activated factor VII

Based on heparin's antithrombin and anti-FXa activity and its in vitro inhibition of activated factor VII (FVIIa) activity, we hypothesized that unfractionated heparin (UFH) may decrease plasma levels of FVIIa in humans. Therefore, 10 healthy young male volunteers received an intravenous UFH infusion over 24 h. Heparin decreased FVIIa levels by 30% (95% CI 14-47%) at 12 h, which was sustained until 24 h. In contrast, neither the substrate pool (i.e. total factor VII) as measured by FVII antigen nor FVII activity were affected by UFH. These results may improve our understanding of the regulation of FVIIa levels and heparin's mode of action.     

The effect of phospholipase C on plasma factor VII

The proposed link between a circulating factor VII-phospholipid complex and the risk of cardiovascular disease (CVD) has stimulated us to investigate the effect of phospholipase C (PLC) on the factor VII (FVII) activity in plasma from healthy individuals. PLC caused a rapid fall in FVII activity which was larger with heparinized than with citrated plasma. EDTA inhibited the PLC effect so emphasizing the involvement of divalent cations. PLC dependent loss of FVII activity varied widely between individuals, showed a highly significant correlation with plasma triglyceride concentrations, and was always greater in post-prandial compared to fasting plasma samples. Experiments using pure recombinant FVIIa and plasma depleted of FVII by adsorption indicated that loss of FVII activity only occurred in the simultaneous presence of absorbed plasma, FVIIa and PLC. Preincubation of PLC with adsorbed plasma before adding FVIIa did not lead to loss of FVII activity. It appears that PLC may act on lipoproteins already bound to FVII, in order to inhibit FVII activity. Other results indicated that competition between different plasma components (lipoproteins) in binding to FVII may govern the extent of the PLC dependent reduction in FVII activity.     

Influence of warfarin therapy on activated factor VII clotting activity.

Oral anticoagulant therapy has been widely employed to prevent and treat a variety of thrombotic disorders, although a new generation of antithrombotic drugs, which offer inhibition of clot-bound as well as fluid-phase thrombin, has been developed and tested in several clinical trials. Although most anticoagulant responses to hydroxycoumarin compounds are well established, there are controversial evidences on their influence on activated factor VII (FVIIa). After analyzing the prothrombin time (PT) (International Normalized Ratio reference range, 0.92-1.08), factor VII clotting activity (FVII:C) (reference range, 75-130 U/dl) and activated factor VII clotting activity (FVIIa:C) (reference range, 30-110 U/l) in 46 consecutive patients on stable warfarin therapy for atrial fibrillation, a consistent trend towards decreased values of both FVII:C and FVIIa:C was observed as PT values increased. At moderate-intensity anticoagulation, with international normalized ratios between 2 and 3, the mean activities of FVII:C and FVIIa:C dropped to 28 U/dl (range, 9-61 U/dl) and 5.8 U/l (range, 1-18 U/l), respectively. Results of our investigations indicate that warfarin therapy decreases FVIIa:C, highlighting the potential benefits of oral anticoagulants in thrombotic disorders and other clinical conditions characterized by increased levels of FVIIa. Owing to the good correlation with FVIIa:C, we also hypothesize that the PT and/or FVII:C might be employed for monitoring recombinant FVIIa therapy.