GPⅡb基因型急性心肌梗死患者介入治疗前后GMP-140、vWF和PAI-1水平

目的：探讨3种血小板膜糖蛋白（GP）Ⅱb HPA-3基因型急性心肌梗死（AMI）患者经皮冠状动脉介入治疗（PCI）前后血浆中血小板α-颗粒膜蛋白（GMP-140）、血管性血友病因子（vWF）和纤溶酶原激活物抑制物-1（PAI-1）变化.方法：对102例AMI患者进行GPⅡb HPA-3基因分型,比较各基因型AMI患者的临床特征,并测定PCI术前、术后即刻及术后24 h3个时间点血浆GMP-140、vWF和PAI-1水平.结果：102例AMI患者中,GPⅡb HPA-3 aa基因型34例,ab基因型38例,bb基因型30例,3种基因型AMI患者的临床特征比较无统计学意义;3种基因型AMI患者GMP-140在PCI术后开始增高（P＜0.01）,术后24 h下降,bb基因组在PCI术后即刻、术后24h较aa、ab基因组均增高（P ＜0.01）;vWF在PCI术后即刻、术后24 h（P ＜0.01）持续增高,但3种基因组间差异无统计学意义;PAI-1在术后即刻、术后24h持续升高,bb基因组在术后即刻、术后24h活性明显高于aa、ab基因组（P＜0.01）.结论：GPⅡb HPA-3基因型AMI患者PCI前后血浆GMP-140、vWF和PAI-1水平发生变化,尤其bb基因型在PCI术后存在更明显的血小板活化.     

Activity of platelet in patients with high level of LDL and the effect of LDL on platelet glycoproteins

OBJECTIVE: To investigate the activity of platelet in patients with high level of low density lipoproteins (LDL) and the effect of LDL on platelet glycoproteins (GP). METHODS: Platelet glycoproteins, whose platelet were activated or unactivated, were measured by flow cytometry. RESULTS: The amount of the platelet membrane GP II b/III a in patients was not significantly different from that of the control when platelet was unactivated (P > 0.05); when platelet was activated by adenosine diphosphate (ADP), the amount of patients' platelet GP II b/III a was increased markedly in comparison with that in the control (P < 0.01). Having been preincubated with LDL, the platelets were activated by ADP and the amounts of GP II b/III a of patients group and the control were all increased obviously as compared with those in the platelets which were not incubated with LDL (P < 0.01; P < 0.01). There was no significant difference of the amount of granule membrane protein-140 (GMP-140) between patients and the control when platelet was activated by thrombin (P > 0.05). Having been preincubated with LDL, platelet was activated by thrombin, the amounts of GMP-140 on patients' and control platelet were all increased markedly (P < 0.01, P < 0.01). CONCLUSION: The activity of platelet in patients with high level of LDL is increased significantly. LDL can increase the expression of platelet glycoproteins.     

定量测定45例血小板膜糖蛋白及其在急性脑梗死中的意义

目的:探讨流式细胞术定量测定血小板膜糖蛋白在急性脑梗死中的临床意义。方法:应用流式细胞仪测定静息状态和TRAP激活的血小板所结合单抗的分子数。结果:急性脑梗死患者血小板静息状态下GPⅢa和GPⅡb/Ⅲa密度[分别为(76.8±14.1)×103/血小板、(73.9±10.5)×103/血小板]高于健康对照组[分别为(55.6±5.8)×103/血小板、(50.5±4.1)×103/血小板,P<0.05],TRAP活化后患者血小板GPⅢa和GPⅡb/Ⅲa密度[分别为(102.6±16.4)×103/血小板、(108.5±16.7)×103/血小板]亦明显高于对照组[分别为(69.7±18.3)×103/血小板、(61.2±5.7)×103/血小板,P<0.01];患者血小板静息状态下,GMP-140表达与对照组比较没有显著性差异(P>0.05),而TRAP激活后患者血小板GMP-140表达[(1.21±0.41)×103/血小板]明显高于对照组[(0.87±0.26)×103/血小板,P<0.05];患者血小板静息和活化状态GPⅠb表达与对照组比较没有显著性差异(P>0.05)。结论:急性脑梗死患者血小板呈较为活化状态,血小板膜糖蛋白测定对病情的分析和疗效的判断有重要的临床参考价值。     

血府逐瘀口服液对冠心病血瘀证患者血液流变学的影响及其与人类血小板抗原3基因多态性的相关性

目的：观察血府逐瘀口服液对冠心病血瘀证患者血液流变学的影响,分析其与血小板膜糖蛋白Ⅱb（glycoproteinⅡb,GPⅡb）人类血小板抗原3（human platelet antigen-3,HPA-3）基因多态性的相关性。方法：筛选符合入选标准的32例冠心病血瘀证患者,进行血瘀证计分并检测血液流变学,采用TaqMan探针技术检测GPⅡb HPA-3基因多态性,血府逐瘀口服液治疗4周后复查血液流变学并进行血瘀证计分。结果：血府逐瘀口服液治疗后冠心病血瘀证患者血液流变学得到改善,含有至少一个突变型基因C的患者治疗前后血液黏度、红细胞变形指数及血瘀证计分与AA型患者比较,差异有统计学意义（P〈0.05）。结论：血府逐瘀口服液能减轻冠心病血瘀证患者的血瘀症状,改善冠心病血瘀证患者的血液流变性,这种改善作用与GPⅡb HPA-3基因多态表型有相关性。     

老年冠心病患者血小板α颗粒膜蛋白-140检测的临床意义

目的 探讨老年冠心病患者血小板α颗粒膜蛋白－140检测的临床意义。方法 采用流式细胞仪（FCM）和单克隆抗体CD62p测定38例稳定性心绞痛（SAP）患者，50例不稳定性心绞痛（UAP）患者和8例急性心肌梗死（AMI)中层得的外周血中血小板α颗粒膜蛋白－140（GMP－140），并随机选择35例体检者作为对照组。结果 老年冠心病患者的GMP－140阳性率与对照组的比较，差异有显著性意义（P＜0．05）。结论 老年冠心病患者血浆GMP－140含量增高，抗血小板活化在冠心病的治疗中有重要意义。     

胰岛素对正常人血小板膜糖蛋白Ⅱb／Ⅲa表达的影响及机制

目的 探讨胰岛素对正常人血小板膜糖蛋白(GP)Ⅱb／Ⅲa表达的影响及其机制。方法 建立全血血小板膜GPⅡb/Ⅲa表达的流式细胞术测定方法，观察胰岛素以及一氧化氮合酶抑制剂硝基左旋精氨酸甲酯(L-NAME)、鸟苷酸环化酶(GC)抑制剂亚甲蓝(MB)分别及与胰岛素联合作用下对血小板膜GPⅡb/Ⅲa表达的影响。结果 (1)胰岛素对正常人静息血小板膜GPⅡb／Ⅲa表达无明显影响；而显著抑制凝血酶、胶原活化的血小板膜GPⅡb／Ⅲa的上调表达，且该效应呈剂量和时间依赖性；(2)LNAME、MB几乎完全阻断胰岛素对血小板膜GPⅡb／Ⅲa表达的抑制效应。结论 胰岛素可通过NOS→NO→GC→GMP通路抑制血小板膜GPⅡb／Ⅲa的表达。     

Effect of Xiaoyu Zhixue tablet on expression of platelet membrane glycoproteins in patients with hemorrhagic thrombopathy

Objective: To observe the effect of Xiaoyu Zhixue tablet (XYZXT) on the expression of platelet membrane glycoproteins in patients with hemorrhagic thrombopathy, and to explore its possible mechanism. Methods; The total of 148 patients with hemorrhagic thrombopathy were randomly divided into two groups, the traditional Chinese medicicne (TCM) group (n = 98) treated with XYZXT and the Western medicine (WM) group (n = 50) treated with adrenosin, vitamins C, K and P, both for 6 months. The therapeutic effect and the recovery rate of platelet aggregation in the two groups were observed. And platelet membrane glycoprotein (GP) Ib/IX, GPII b/IIIa complexes, GPIb, GPIIb, GPIIIa and P-selectin were analyzed by flow cytometry in both groups before and after treatment and also in 34 normal healthy subjects.Results: The total effective rate of hemostasis was 89.8% in TCM group and 54.0% in the WM group (X ²=45.83,P<0.01), and the recovery rate of platelet aggregation was 72.4% and 4.0% respectively (X ² = 62.06,P<0.01). The fluorescence intensity of GP I b/IX, GP II b/III a complexes, GPIb, GP IIIa and P-selectin were lower in both groups before treatment than those in the healthy subjects. Expression of above-mentioned marks was elevated in TCM group after 6 months’ therapy, which was insignificantly different as compared with the healthy subjects (P>0.05) and higher than those in the WM group (P<0.05).Conclusion: One of the mechanisms in treating hemorrhagic thrombopathy with XYZXT is that it could regulate the expression of GP I b/IX, GPII b/IIIa complexes, GP I b, GPIIIa and P-selectin at the level of receptor protein. Supported by the National Natural Science Foundation of China (No. 30171194)     

老年脑梗死患者急性期血小板活化的检测及临床意义

目的比较老年急性脑梗死(ACI)患者与健康老年人血小板功能的不同。方法采用玻璃球旋转法、比浊法和流式细胞术检测36例ACI患者和19例对照人群的血小板黏附率(PAgT)以及血小板膜糖蛋白(GMP)CD41、CD62p和CD63的阳性表达率。结果老年ACI患者组PAdT、PAgT、CD41、CD62p和CD63的检测值,分别为31%±13%、40%±14%、12.34%±3.31%、7.82%±2.59%、21.34%±6.63%;对照组上述各项检测值,分别为22%±9%、27%±13%、8.98%±2.5%、4.87%±1.79%、17.87%±4.45%。结论老年ACI患者血小板处于激活状态,血小板黏附、聚集、释放等功能增强,血小板活化与老年ACI的发病机制有关。     

Differential gene expression profiles in coronary heart disease patients of blood stasis syndrome in traditional Chinese medicine and clinical role of target gene

Objective:To investigate the differential gene expression profiles in coronary heart disease(CHD) patients of blood-stasis syndrome(BSS) by oligonucleotide microarray technique,and the clinical significance of target gene.Methods:Subjects were assigned to CHD patients with BSS(n=8),CHD patients without BSS (n=8),and BSS patients without CHD(n=8) based on coronary angiography and the diagnostic criteria of BSS. The sex- and age-matched healthy volunteers(n=8) were enrolled as the control group.Venous bloo...     

介入治疗对老年急性冠脉综合征患者血小板活化和炎症因子的影响

目的 探讨经皮冠状动脉介入治疗（PCI）对老年急性冠脉综合征(ACS)患者血小板活化和炎症因子的影响。方法68例经PCI治疗的老年ACS患者,在术前、术后次日、术后1周及术后4周清晨分别空腹抽肘静脉血测定P选择素（CD62P）、GPⅡb/Ⅲa受体复合物、血浆纤维蛋白原(fg)及血清高敏反应蛋白(hs-CRP)水平。并选择健康体检者44名作正常对照组。结果 CD62p、GPⅡb/Ⅲa、fg、hs-CRP水平ASC患者PCI术前、术后均较正常对照组增高（P<0.05）,CD62p、GPⅡb/Ⅲa、fg、hs-CRP水平术前、术后在不同冠脉病变组间无统计学差异（P>0.05）。结论 老年ACS患者存在血小板活化和炎症反应,CD62p、GPⅡb/Ⅲa、fg、hs-CRP水平在术后4周内增高,术后4周内的抗血小板及抗凝治疗对防治PCI术后冠脉内血栓形成及再狭窄至关重要。     

GP IIb三种基因多态性之间的关系及GP IIb T13959 G在血小板输注耐受中的作用

目的:研究血小板膜糖蛋白Ⅱb (GP Ⅱb)三种基因多态性之间的关系及GP Ⅱb T13959 G在血小板输注耐受中的作用.方法:聚合酶链反应-单链构型多态性分析检测110例正常人GP Ⅱb基因第26和30外显子(Exon 26, Exon30)及21内含子(Intron21)基因多态性,进行基因序列分析,研究这些基因多态性是否存在连锁关系;应用Fok1酶切法对147例血液病人人类血小板抗原-3(human platelet antigen-3, HPA-3)基因进行分型,并与110例正常人进行比较;将接受单采血小板输注的44例血液病人随机分为HPA-3同型输注组和对照组,血小板输注3次以后检测血小板抗体.结果:正常人GP Ⅱb存在基因多态性,表现分别为gDNA 13 959位点T→G,16 977位点C→T,11 996～12 004位点9个碱基缺失,且三者具有同步性.Fox1酶切分析结果表明,正常人和血液病人的HPA-3a基因频率分别为83.6%(92/110)和81.9%(119/147),HPA-3b基因频率分别为16.4%(18/110)和19.1%(28/147),两者之间差异无统计学意义(P＞0.05).血液病人接受单采血小板输注后, HPA-3同型输注组血小板抗体阴性,而对照组有2例存在血小板抗体,其中1例为HPA-3a抗体.结论:(1)GP Ⅱb基因存在gDNA 13 959位点T→G,16 977位点C→T,11 996～12 004位点9个碱基缺失,这3个位点的多态性可能存在完全连锁关系;(2)血液病人与正常人的HPA-3基因频率相同;(3)HPA-3系统可能是我国人群产生血小板输注耐受的原因之一.     

人血小板功能抗体及其片段的识别

目的研究特发性血小板减少性紫癜（ITP）患者血小板膜糖蛋白（GP）特异性IgG抗体及其片段的免疫活性及对血小板聚集功能的影响.方法用改良单克隆抗体特异性俘获血小板抗原技术（MAIPA）检测84例慢性ITP患者血浆中抗GPⅡb/Ⅲa、GPⅠb/Ⅸ及GPⅥ自身抗体,比浊法血小板聚集试验筛选出自身抗体阳性并能抑制血小板聚集的患者,用蛋白A柱纯化其血浆IgG抗体并用胃蛋白酶制备F（ab＇）2片段.检测纯化的IgG抗体及其酶切片段与血小板GP的结合活性及对正常人血小板聚集功能的影响.结果 （1）84例ITP患者血浆中,48例（57.1%）抗GPⅡb/Ⅲa和/或GPⅠb/Ⅸ和/或GPⅥ自身抗体阳性,其中7例（14.6%）明显抑制了二磷酸腺苷、瑞斯托霉素或胶原诱导的血小板聚集;（2）纯化的IgG及F（ab＇）2片段具有与相应血小板GP的结合活性;（3）4例患者纯化IgG及F（ab＇）2片段具有抑制血小板聚集的功能.结论 F（ab＇）2片段是IgG自身抗体的功能片段,它不但保留了良好的抗原结合活性且可部分抑制血小板聚集功能,为人源化血小板糖蛋白特异性抗体的制备奠定了基础.     

血管内皮损伤和血小板活化与急性冠脉综合征的相关性研究

目的:探讨血管内皮损伤和血小板活化与急性冠脉综合征(ACS)的相关性。方法:选择我院收治的急性冠脉综合征患者40例作为研究对象,记为研究组,35例稳定型心绞痛患者作为对照1组,并选取同期门诊体检健康志愿者35例作为对照2组。采用流式细胞仪检测CD62p、CD63、GPⅡb/Ⅲa受体复合物,采用散射比浊法测定纤维原蛋白(FIB-C),采用ELASA测定假血友病因子vWF、内皮素1(ET-1),采用酶法测定NO,采用彩色多谱勒测定内皮依赖性血管舒张功能,并作以比较。结果:(1)研究组vWF、ET-1较对照1组、对照2组明显升高,(P<0.05);NO、FMD较对照1组、对照2组明显降低(P<0.05)。(2)研究组CD62p、CD63、GPⅡb/Ⅲa受体复合物以及FIB-C等反应血小板活化功能的指标明显高于对照1组和对照2组,(P<0.05)。结论:急性冠脉综合征患者反应血管内皮损伤和血小板活化的指标发生明显变化,说明血管内皮损伤和血小板活化与急性冠脉综合征有密切关系。     

心肌缺血时血小板功能变化的机制

研究了冠心病人以及实验大鼠心肌缺血期间血小板功能变化的机制。利用血小板聚集仪检测血小板聚集率（PAgR）。应用CD41,CD62和标记第二抗体FITC在流式细胞仪上检查了GPⅡb/Ⅲa和GMP-140表达的变化。利用荧光血小板聚集仪测定了血小板ATP的释放量;利用血小板离子钙聚集仪检测胞浆内钙含量。结果表明缺血时PAgR的增高是由于血小板表面的GPⅡb/Ⅲa受体表达增加所致。缺血期间GMP-140表达的增加表明心肌缺血时血小板易于被激活,并且黏滞性增高。心肌缺血时ATP释放量也明显增加。在血小板ATP释放反应和血小板聚集率之间有正反馈相互作用关系。心肌缺血能显著引起胞浆内钙含量的增高,后者是导致血小板功能异常变化的重要细胞内机制。     

心肌梗死患者血清脑钠肽、血液流变学及血小板活化指标变化的研究

目的探讨心肌梗死患者血清脑钠肽(BNP)、血液流变学及血小板活化指标的变化规律。方法选择2010年9月至2012年4月于河北北方学院附属第三医院进行治疗的37例心肌梗死患者作为观察组,同期选择在本院进行体检的37名健康体检者作为对照组,检测两组的血清BNP、血液流变学及血小板活化指标并进行比较。结果观察组的血清BNP、血浆黏度、全血黏度、红细胞聚集及刚性指数、CD62p、血小板膜糖蛋白纤维蛋白原受体、CD61、GPⅡb/Ⅲa、血小板α颗粒膜蛋白140水平均明显高于对照组,且Killip分级Ⅲ～Ⅳ级患者高于Ⅰ～Ⅱ级患者(均P<0.05)。结论心肌梗死患者血清BNP、血液流变学及血小板活化指标均呈现升高的趋势,且严重程度对其升高程度有一定影响。     

Study on blood coagulant/fibrinolytic activity at plasma and monocytic levels in coronary heart disease patients with blood-stasis syndrome of traditional chinese medicine

Study on Blood Coagulant/Fibrinolytic Activity at Plasma and Monocytic Levels in Coronary Heart Disease Patients with Blood-Stasis Syndrome of Traditional Chinese Medicine@吕 中 @施赛珠     

Correlation of Lower Concentrations of Hydrogen Sulfide with Activation of Protein Kinase CβII in Uremic Accelerated Atherosclerosis Patients

Background:

Hydrogen sulfide (H₂S) plays a protective role in chronic hemodialysis (CHD) patients. In this study, we further investigate the relationship between H₂S and conventional protein kinase CβII (cPKCβII) in CHD patients with uremic accelerated atherosclerosis (UAAS).

Methods:

A total of 30 healthy people, 30 CHD patients without AS and 30 CHD patients with AS (CHD + AS) were studied. Plasma H₂S was measured with a sulfide sensitive electrode, and cPKCβII membrane translocation was detected by Western blotting.

Results:

Plasma H₂S in CHD + AS group was significantly lower than that in CHD patients. cPKCβII membrane translocation in CHD + AS group increased significantly compared with CHD group. Plasma H₂S concentration was negatively correlated with cPKCβII membrane translocation in CHD + AS patients.

Conclusions:

These findings suggest a possible linkage between H₂S metabolism and cPKCβII activation, which may contribute to the development of UAAS in CHD patients.     

Effect of xuefu zhuyu decoction on the function of platelets and human umbilical vein endothelial cell

Objective: To explore the effect of Xuefu Zhuyu Decoction (XZD) on molecular expression of platelets glycoprotein II b/III a complex (GP II b/III a) and thrombomodulin (TM) of human umbilical vein endothelial cells.Methods: Platelets and human umbilical vein endothelial cells were incubated with XZD of different concentration. GP II b/III a and TM were evaluated by radioimmunoassay.Results: XZD in 40 mg/ml and 80 mg/ml could obviously inhibit the adenosine diphosphate induced GII b/III a complex expression, the molecular number being 52900 ± 8445 and 52095 ± 6345 respectively, and there was significant difference as comparing with that in the control group ( P < 0.05, P<0.01). But XZD didn’t show any influence on the molecular expression of TM in human umbilical vein endothelial cells.Conclusion: XZD could inhibit the adenosine diphosphate induced activation of platete through blocking the exposure of GII b/III a a complex.     

冠心病病人血浆vWF、GMP-140及Fg变化的临床分析

目的：探讨冠心病（CHD）病人血浆vWF、GMP-140及Fg在CHD发痛中的作用。方法：将CHD病人分为3组，观察各组血浆vWF、GMP-140及Fg的含量，与对照组比较。培果：CHD病人血浆vWF、GMP-140厦Fg与对照组比较有显著性差异（P＜0．05）；AMI上述指标明显增高（P＜0．01）。结论：CHD存在明显的高凝状态，vWF、GMP-140及Fg可预：则血栓形成的危险性。     

冠状动脉性心脏病患者血小板表面膜糖蛋白的表达

目的研究冠状动脉性心脏病（coronary heart disease,CHD）患者血小板表面纤维蛋白原结合位点（PAC-1）、膜糖蛋白P-选择素（CD62P）、GPIIb11Ia（CD41a）和GPⅠb／Ⅸ（CD42b）的变化及临床意义。方法用流式细胞术（FCM）观察53例CHD患者及30名正常人PAC-1、CD62P、CD41a和CD42b表达。结果CHD患者PAC-1、CD62P、CD41a异常增高（P〈0．01）,CD42b降低（P〈0．05）。结论CHD患者伴有血小板活化,存在高凝状态病理基础,容易导致血栓的形成。