血清MMP-9与NSE在ACI患者中检测的意义

目的探讨血清基质金属蛋白酶-9(MMP-9)与神经烯醇化酶(NSE)水平检测在急性脑梗死(ACI)患者中的临床价值。方法分析2014-12-2015-09在开封市中心医院神经内科接受诊治的94例ACI患者的临床资料。根据病灶面积的大小分为A组(大面积脑梗死,21例)、B组(中等面积脑梗死,46例)和C组(小面积脑梗死,27例)。另选取同期健康体检的92例健康者为对照组。比较2组基线资料、血清MMP-9与NSE的表达水平,并分析ACI患者血清MMP-9与NSE的关系。结果ACI组血清MMP-9与NSE浓度明显较对照组高(P0.05)。其中,ACI组血清MMP-9与NSE水平随着梗死面积的增大而逐渐上升,A、B、C 3组组间比较有显著性差异(P0.05)。ACI患者血清MMP-9与血清NSE水平存在正相关性(r=0.568,P0.05)。结论血清MMP-9与NSE水平的检测可用于判断ACI患者病情严重程度及其预后情况,具有重要的临床价值。     

血清S-100β神经元特异性烯醇化酶水平与脑卒中患者神经功能损害程度的相关性研究

目的探讨血清S-100β、神经元特异性烯醇化酶(NSE)与脑卒中患者神经功能损害程度的相关性。方法选取2017-02—2020-02开封市人民医院治疗的115例急性缺血性脑卒中患者为观察组,同时选取健康志愿者50例为对照组,检测2组血清S-100β、NSE水平。结果观察组血清S-100β、NSE水平明显高于对照组(P<0.05);观察组中大面积梗死患者S-100β、NSE水平高于腔隙性梗死和小面积梗死患者(P<0.05);观察组美国国立卫生研究院卒中量表(NIHSS)评分≥31分患者血清S-100β、NSE水平高于NIHSS评分16~30分和≤15分患者(P<0.05);血清S-100β、NSE水平与梗死面积呈正相关(P<0.05),与NIHSS评分亦呈正相关(P<0.05)。结论急性缺血性脑卒中患者血清S-100β、NSE水平升高,与梗死面积、神经功能损害程度存在正相关关系。     

急性脑梗死患者血清MMP-9与S-100B蛋白水平变化及临床意义

目的探讨血清基质金属蛋白酶-9(MMP-9蛋白)与S-100B蛋白在急性脑梗死(ACI)病理过程中的变化及临床意义。方法采用酶联免疫吸附(ELISA)法检测65例ACI患者急性期及恢复期血清MMP-9和S-100B蛋白浓度并与35例健康对照比较;比较不同梗死面积的ACI患者在急性期血清MMP-9和S-100B浓度的差异;分析急性期ACI患者MMP-9和S-100B的相关性。结果 ACI患者急性期血清MMP-9和S-100B蛋白浓度均明显高于恢复期及对照组(P0.01);急性期脑梗死面积较大的患者血清MMP-9和S-100B蛋白浓度高于脑梗死面积较小的患者(P0.01);急性期MMP-9与S-100B蛋白呈正相关性(r=0.413,P0.01)。结论 ACI患者血清MMP-9和S-100B蛋白浓度随病程、梗死面积的大小而变化,并且在急性期时MMP-9和S-100B表达呈正相关。联合检测ACI患者血清MMP-9和S-100B蛋白浓度对ACI的诊断、病情监测和疗效观察具有重要参考作用。     

S-100β蛋白和神经元特异性烯醇化酶在急性脑梗死中的检测意义

目的 探讨S-100β蛋白和神经元特异性烯醇化酶在急性脑梗死中的检测意义.方法 选择我院2009-04-2011-04急性脑梗死患者100例作为观察组,其中轻型31例,中型42例,重型27例;腔隙性梗死30例,小面积梗死45例,大面积梗死25例.选择同期我院体检健康者50例作为对照组.检测观察组和对照组S-100β蛋白和神经元特异性烯醇化酶水平.结果 观察组S-100β蛋白和神经元特异性烯醇化酶水平显著高于对照组,差异有统计学意义(P<0.05);随着脑梗死面积增加,S-100β蛋白和神经元特异性烯醇化酶水平显著升高,差异有统计学意义(P<0.05);随着病情严重程度增加,S-100β蛋白和神经元特异性烯醇化酶水平显著升高,差异有统计学意义(P<0.05).结论 S-100β蛋白和神经元特异性烯醇化酶联合检测有助于了解急性脑梗死患者梗死面积大小及判断病情严重程度,为临床诊断和治疗提高参考.     

血清NSE、Hcy及H-FABP检测在急性脑梗死诊断中的应用价值

目的探讨血清神经元特异性烯醇化酶(NSE)、同型半胱氨酸(Hcy)及心型脂肪酸结合蛋白(H-FABP)检测在急性脑梗死(ACI)诊断中的应用价值。方法分析100例ACI患者(观察组)和100例健康体检者(对照组)血清NSE、Hcy及H-FABP水平差异,对比不同病灶直径及神经损伤程度ACI患者血清NSE、Hcy及H-FABP水平;分析各指标对ACI的诊断价值。结果观察组血清NSE、Hcy及H-FABP水平均显著高于对照组(P0.05)。随病灶直径的增加和神经损伤程度的加重,血清NSE、Hcy及H-FABP水平也显著升高(P0.05)。ACI患者血清NSE、Hcy及H-FABP水平与病灶直径及患者NIHSS评分均呈显著正相关(P0.05)。血清NSE、Hcy及H-FABP诊断ACI的ROC曲线下面积分别为0.913、0.872、0.822,其诊断敏感度为72.0%、73.0%、77.0%,特异度为97.0%、88.0%、71.0%。结论血清NSE、Hcy及H-FABP水平随ACI患者病灶直径及神经损伤程度的加重而升高,三者可作为ACI临床诊断和评估病情的重要实验室指标。     

急性脑梗死患者血小板微颗粒水平的研究

目的研究急性脑梗死患者的血小板微颗粒(PMP)水平及其临床意义。方法检测并比较80例急性脑梗死患者及80例健康人PMP水平,比较不同面积、不同TOAST分型脑梗死患者PMP水平的差异,检测脑梗死治疗后不同时期PMP的动态变化。结果与健康对照组比较急性脑梗死患者PMP水平较高,且大面积脑梗死患者PMP水平较其他脑梗死患者更高。不同TOAST分型脑梗死患者PMP水平差异显著。治疗后脑梗死患者PMP水平呈动态下降。结论急性脑梗死患者存在血小板活化,检测血液PMP水平将有助于急性脑梗死的诊断和治疗。     

可溶性CD40L、MMP-9与颈动脉粥样硬化及急性脑梗死的关系

目的探讨可溶性CD40L(sCD40L)、基质金属蛋白酶-9(MMP-9)与颈动脉粥样硬化及急性脑梗死(ACI)的关系。方法选取首次发病的ACI患者90例和健康对照组30例,采用双抗体夹心酶联免疫吸附试验(ELISA)测定血清sCD40L、MMP-9的水平。应用颈动脉超声检测颈动脉内膜状况。比较不同程度颈动脉粥样硬化及不同面积脑梗死患者血清sCD40L、MMP-9的水平变化,并对所有脑梗死患者进行神经功能缺损评分。结果 ACI患者血清sCD40L、MMP-9的水平显著高于对照组(P0.01);大梗死组血清sCD40L、MMP-9水平高于中、小梗死组,中梗死组高于小梗死组,差异均有统计学意义(均P0.01);随着颈动脉粥样硬化程度加重,脑梗死病情越重及脑梗死面积越大,血清sCD40L、MMP-9的水平也越高;血清sCD40L与MMP-9的水平呈正相关(r=0.887,P0.01)。结论 ACI患者血清sCD40L、MMP-9水平可以反映颈动脉粥样硬化斑块的性质和稳定性、脑梗死面积与病情的严重程度;CD40-CD40L系统可能通过上调MMP-9导致颈动脉粥样硬化斑块不稳定。     

急性脑梗死患者S-100、NSE的测定及意义

目的研究急性脑梗死(ACI)患者血清S-100、NSE含量的变化及意义。方法分别采用放免法、ELISA法测定60例ACI患者治疗前、后血清S-100、NSE的含量。结果ACI患者治疗前血清S-100、NSE浓度明显高于对照组(P<0.05);二者治疗前、后比较有亦有显著性差异(P<0.05)。结论ACI患者血清S-100、NSE的变化可反映患者神经胶质细胞、脑神经细胞的损害情况和功能状态。     

急性脑梗死患者血浆S-100蛋白含量的动态变化及其临床意义

目的研究急性脑梗死（ACI）患者血浆S-100蛋白含量的动态变化及其临床意义。方法采用酶联免疫吸附法（ELISA）检测30例ACI患者病后1d、2d、3d、5d、7d、10d的血浆S-100蛋白浓度,并与30名健康人作对照;在发病1d、10d进行中国卒中量表（CSS）评分。结果与对照组相比,ACI患者发病1d血浆S-100蛋白含量即明显升高（P〈0.01）,2~3d达到峰值,5d下降,7d降至正常;大面积脑梗死患者血浆S-100蛋白含量高于中、小面积梗死患者,中面积梗死又高于小面积梗死患者（P〈0.05~0.01）。发病后1-5d血浆S-100蛋白浓度与CSS呈正相关（r=0.468-0.642,均P〈0.01）。结论ACI患者血浆S-100蛋白含量的变化可以反映疾病的严重程度,对指导治疗、评估预后有重要意义。     

进展性脑梗死患者血清C-反应蛋白、S-100β蛋白水平变化及意义

目的探讨进展性脑梗死(PCI)患者血清C-反应蛋白(CRP)与S-100β蛋白(S-100β)水平的变化及意义。方法100例急性脑梗死(ACI)患者根据发病后7 d内神经功能缺损量表评分(SSS)的变化分为PCI组(38例)和非PCI(NPCI)组(62例)。检测两组发病后第1 d、3 d、7 d、14 d血清CRP与S-100β的水平,并与健康对照组比较。结果与NPCI组比较,PCI组发病后各时间点血清CRP及S-100β水平明显升高(均P<0.01);与健康对照组比较,NPCI组血CRP与S-100β水平在发病第1~7 d明显增高(均P<0.01),14 d恢复正常水平。ACI组血清S-100β与CRP水平呈正相关(r=0.856,P<0.01);PCI组发病第1 d血清CRP和S-100β水平与分别第7 d SSS正相关(r1=0.695,r2=0.731;均P<0.05)。结论ACI患者发病早期血清CRP及S-100β水平增高与其病情及PCI的发生相关。     

Circulating tissue factor-exposing microparticles

Upon stimulation or apoptosis, eukaryotic cells shed membrane vesicles of submicron size. These so-called microparticles (MPs) are detected and characterized based on the exposure of antigens characteristic of their respective parental cells and on the increased distribution of negatively charged phospholipids to the outer membrane layer. Among the various hypothesized functions of MPs in both health and disease, one of the most studied is their possible role in hemostasis and thrombosis. In this context, special attention is paid to tissue factor (TF) exposed on a variety of MPs. MPs may have outstanding functional because of their ability to display "active" TF due to an abundance of negatively charged phospholipids on their surface. The rapid accumulation of TF-bearing MPs (TF+MPs) in a developing thrombus as well as the increased numbers and thrombogenic activity of TF+MPs in prothrombotic disorders indicate an important role in the pathogenesis of thrombosis. Nevertheless, isolation, quantification and antigenic characterization of TF+MPs proved challenging and a lively scientific debate is ongoing with respect to a reliable method to determine the cellular source of MP in vivo. Standardization of preanalytical procedures and development of more sensitive technologies are needed to improve our current understanding of the role of circulating TF+MPs in thrombosis.     

Tissue factor microparticles and haemophilia

Treatment choices for haemophilia patients with inhibitors are suboptimal. Tissue factor-bearing microparticles home to thrombi in a cell adhesion molecule-dependent fashion. Their potential utility as a procoagulant is discussed along with the challenges of evaluating this approach in mouse models of haemophilia.     

Coagulation activities of plasma microparticles

An evaluation of the effect of plasma microparticles (MP) on in vitro coagulation has been undertaken using platelet rich (PRP), platelet poor (PPP) and platelet free (PFP) plasmas prepared by differential centrifugation. MP provide coagulant material which shortens the activated partial thromboplastin time (APTT) and dilute simplastin time (DSTT) which is different from that contributed by commercial phospholipid preparations. The amount of platelet factor three (PF3) available in plasma is directly correlated with the centrifugal force used in its preparation and is present in large amounts in the MP pellet remaining after preparation of PFP. Factor VIII (F.VIII:C) and von Willebrand factor (vWf) were associated with the MP fraction but could be separated from MP on sucrose gradients. The effect of MP on the APTT was independent of the F.VIII:C/vWf and was not solely due to their PF3 content. Plasma prepared for routine coagulation assays contains MP which contribute to the APTT and DSTT and should be considered in their assessment. High speed centrifugation of plasma reduces the F.VIII:C/vWf:Ag/RCoF levels and this may contribute to losses of these proteins during preparative procedures utilising high speed centrifugation.     

Formation of procoagulant microparticles and properties

The platelet procoagulant response consists of providing a catalytic surface where vitamin K-dependent clotting factors can interact with cofactors to form the characteristic enzyme complexes of the cascade culminating in the generation of sufficient thrombin for effective hemostasis. The essential element allowing such a local concentration is the anionic aminophospholipid phosphatidylserine, sequestered in the inner leaflet of the plasma membrane of resting cells but swiftly translocated to the outer leaflet after stimulation. Phosphatidylserine egress is followed by the shedding of membrane fragments, the so-called microparticles or microvesicles, also endowed with procoagulant properties more particularly when they harbor tissue factor, the major initiator of blood coagulation reactions. Furthermore, because microparticles hijack a number of membrane and cytoplasmic components from the cells they derive, they can elicit various responses in proximal or remote cells they interact with and can therefore be viewed as intercellular “macromessengers“. Although several regulatory mechanisms have been proposed, the main actors responsible for the whole process of phosphatidylserine transmembrane redistribution and subsequent microparticle release remain to be identified.     

Platelet-derived microparticles - an updated perspective

Platelet-derived microparticles (PMP) are a heterogeneous population of vesicles (< 1 mm) generated from the plasma membrane upon platelet activation by various stimuli. They are a discrete population differing from the exosomes which originate from the intracellular multivesicular bodies. PMP also differ from the microparticles derived from megakaryocytes despite the presence of several identical surface markers on the latter. The molecular properties and the functional roles of the PMP are beginning to be elucidated by the rapidly evolving research interest, but novel questions are simultaneously raised. This updated perspective discusses the most recent highlights in the PMP research in context with the methodological problems and the paradoxical role of the PMP in health and disease.     

Analysis of tissue factor positive microparticles

There has recently been intense interest in the clinical measurement of tissue factor (TF)-positive microparticles (MPs) in clinical disease states. This interest has been driven by the demonstration of an putative role for circulating TF-positive MPs in animal models of thrombus propagation. Both immunological and functional assays for MP-TF have been described. While each approach has its own advantages and drawbacks, neither has yet been truly established as the ‘gold standard’. Heterogeneity of TF-bearing MPs, such as the variable co-expression of surface phosphatidylserine, may determine not only their procoagulant potential, but also additional properties including rate of clearance from the circulation.     

Involvement of microparticles in diabetic vascular complications

Type 2 diabetes mellitus (T2DM) is associated with increased coagulability and vascular complications. Circulating microparticles (MPs) are involved in thrombosis, inflammation, and angiogenesis. However, the role of MPs in T2DM vascular complications is unclear. We characterised the cell origin and pro-coagulant profiles of MPs obtained from 41 healthy controls and 123 T2DM patients with coronary artery disease, retinopathy and foot ulcers. The effects of MPs on endothelial cell coagulability and tube formation were evaluated. Patients with severe diabetic foot ulcers expressed the highest levels of MPs originated from platelet and endothelial cells and negatively-charged phospholipid-bearing MPs. MP coagulability, calculated from MP tissue factor (TF) and TF pathway inhibitor (TFPI) ratio, was low in healthy controls and in diabetic retinopathy patients (<0.7) but high in patients with coronary artery disease and foot ulcers (>1.8, p≥0.002). MPs of all T2DM patients induced a more than two-fold increase in endothelial cell TF (antigen and gene expression) but did not affect TFPI levels. Tube networks were longest and most stable in endothelial cells that were incubated with MPs of healthy controls, whereas no tube formation occurred in MPs of diabetic patients with coronary artery disease. MPs of diabetic retinopathy and diabetic foot ulcer patients induced branched tube networks that were unstable and collapsed over time. This study demonstrates that MP characteristics are related to the specific type of vascular complications and may serve as a bio-marker for the pro- coagulant state and vascular pathology in patients with T2DM.     

Proteomics of microparticles after experimental pulmonary embolism

Introduction

Microparticles (MPs) are small fragments of apoptotic or activated cells that may contribute to pathological processes in cardiovascular diseases. In studies of MPs in clinical cohorts, it is unclear if observed changes in MP composition are a cause or a result of the cardiovascular disease being studied. The present studies employed a well-characterized rat model of experimental pulmonary embolism (PE) to determine if there were changes in MP characteristics as a result of pulmonary vascular occlusion.

Methods

PE was produced by infusing 25 μm polystyrene microspheres into the jugular vein of anesthetized rats. MPs were isolated by differential centrifugation of arterial blood 18 hr after PE. Proteins were separated by 1D gel electrophoresis and identified from tryptic digests by ultraperformance liquid chromatography (UPLC) coupled with tandem mass spectrometry. Statistical analysis was conducted using the Power Law Global Error Model (PLGEM). Changes in two proteins were confirmed by Western blot.

Results

Experimental PE produced pulmonary hypertension, mild systemic hypotension, hypoxia, hypercapnia and lactic acidosis. MPs showed significant elevation in proteins involved in clotting (fibronectin precursor, fibrinogen alpha, beta and gamma and von Willebrand factor) and several macroglobulin proteins, such as alpha-2-macroglobulin precursor compared with vehicle-treated control rats. Consistent with recent observations of hemolysis in PE, haptoglobin precursor protein, a major protein of hemoglobin clearance, decreased significantly in the PE animals. Plasma d-Dimer concentrations were significantly elevated, indicating that experimental PE produced a pro-coagulant state.

Conclusions

These findings suggest that experimental PE produced significant, changes in MP characteristics to a prothrombotic phenotype.