川崎病患儿血小板CD61和CD62p的变化及其临床意义

目的测定血小板CD61和CD62p的阳性率,探讨并总结川崎病病程中血小板活化标志物CD62p的阳性表达率的变化与临床诊断、疗效判定及冠状动脉异常之间的关系。方法应用流式细胞仪检测25例川崎病患儿和16例正常对照组急性期血小板CD61和CD62p阳性表达率。结果川崎病患儿全血CD61、CD62p阳性表达率较正常对照儿明显增高,具有统计学意义;合并冠状动脉异常者CD62p阳性表达率增高更加显著,CD62p阳性表达率增高具有统计学意义;CD62p的阳性表达率经丙种球蛋白和阿司匹林治疗后能明显减低,但仍高于正常对照儿,具有统计学意义。结论在川崎病免疫性血管炎的病理损害过程中,CD61和CD62p均显著增高,丙种球蛋白和阿司匹林治疗后能抑制KD患儿血小板膜糖蛋白CD62p的高表达,降低血小板的活化过程：川崎病病程过程中动态检测血小板活化标志物CD62p阳性表达率,对川崎病的诊断和疗效判定具有比较重要的临床意义,对预测冠状动脉异常是否发生更具有价值。     

辛伐他汀对糖尿病大鼠血脂和血小板膜糖蛋白表达水平的影响

目的：探讨羟甲基戊二酸单酰辅酶A （HMG-CoA）还原酶抑制剂辛伐他汀对2型糖尿病大鼠血脂和血小板膜糖蛋白表达的影响。方法雌性Wistar大鼠腹腔注射链脲佐菌素（STZ）建立糖尿病模型,造模成功后将其随机分为糖尿病组、辛伐他汀组[辛伐他汀混悬液灌胃,20 mg/（kg ＆#183;d）,共用12周]各10只,另取10只健康大鼠设为对照组,12周后测定及比较各组血脂及血小板膜糖蛋白CD62p、CD63、CD61、CD41的表达。结果糖尿病组总胆固醇、甘油三酯和LDL水平高于对照组,而HDL的水平低于对照组（P＜0.01）;辛伐他汀组总胆固醇、甘油三酯和LDL水平低于糖尿病组,而HDL水平高于糖尿病组（P＜0.01）;糖尿病组血小板膜糖蛋白CD62p和CD63水平较对照组高（P＜0.01）,辛伐他汀组的CD62p和CD63水平较糖尿病组低（P＜0.01）;3组血小板膜糖蛋白CD61、CD41水平比较差异无统计学意义（P＞0.05）。结论辛伐他汀能调节糖尿病大鼠的血脂水平,能降低血小板膜糖蛋白CD62p和CD63的水平,抑制血小板的聚集。     

血细胞分离机对血小板膜糖蛋白影响的观察

目的　了解血小板经血细胞分离机单采后膜糖蛋白的变化。方法　利用流式细胞仪及CD41a、CD41b、CD42a、CD61、CD62 p、PAC 1单克隆抗体对 2 1份机采血小板前后的膜糖蛋白表达率和平均荧光强度 (meanfluores cenceintension ,MFI)进行检测。结果　血小板经血细胞分离机单采后 ,CD41a、CD41b、CD42a的阳性表达率和MFI、CD61的阳性表达率和CD62 p、PAC 1的MFI与采集前比较无明显变化 (均为P >0 .0 5 ) ;CD6 2 p、PAC 1的阳性表达率增高 (均为P <0 .0 5 ) ,CD61的MFI降低 (P <0 .0 1)。结论　血小板经血细胞分离机单采后仍保存了膜糖蛋白的完整性 ,虽有一定比率的血小板活化 ,但MFI无明显改变     

2型糖尿病下肢动脉硬化病变部位和程度与血小板膜糖蛋白表达的相关性研究

目的:探讨血小板膜糖蛋白在2型糖尿病下肢动脉硬化闭塞症(ASO)患者不同部位和不同严重程度的表达及其相关性。方法:80例2型糖尿病合并下肢动脉硬化闭塞症患者,分别检测血小板膜糖蛋白CD62p、CD63、CD61、CD41a、CD42b、纤维蛋白原及血小板黏附、聚集试验,根据影像学资料和临床踝肱指数(ABI)检查区分不同部位和严重程度,并比较其差异。结果:2型糖尿病ASO患者髂股动脉端狭窄闭塞患者的血小板膜糖蛋白CD62p、CD63较腘动脉以下闭塞患者明显升高,CD62p、CD63与ABI无相关性。结论:检测血小板膜糖蛋白对于判断2型糖尿病病变部位有一定临床意义。     

氯吡格雷对急性脑梗死患者血小板膜糖蛋白、血小板聚集率的影响

氯吡格雷是一种血小板高聚集抑制剂,它通过抑制ADP从而干预了纤维蛋白原与血小板膜的结合,具有不可逆的血小板抑制作用,近年来广泛用于急性缺血性脑梗死的治疗。CD62p和CD63是血小板胞浆内仅颗粒上的膜糖蛋白及溶酶体膜糖蛋白,均为活化血小板的膜糖蛋白,成为血小板活化的标志。CD62p是目前最具有特征性的血小板活化的分子标志物,CD63被认为是一种比CD62p更为敏感的血小板活化标志物忙。近年来对于血小板活化功能亢进与缺血性脑血管病的关系日益受到重视H0。本研究以急性脑梗死为研究对象,通过观察氯吡格雷对急性脑梗死患者血小板膜糖蛋白CD62p、CD63和血小板聚集率的影响及临床疗效观察。现报告如下。     

2型糖尿病患者血小板CD41、CD61、CD62p的表达及意义

目的研究2型糖尿病患者血小板膜糖蛋白变化与血管病变的关系及其影响因素。方法用流式细胞术测定35例2型糖尿病有血管病变患者、32例2型糖尿病无血管病变患者及31例健康者血小板膜糖蛋白CD41、CD61、CD62p的表达。结果2型糖尿病伴血管病变组CD62p显著高于无血管病变组及健康对照组(P<0.001),无血管病变组与正常对照组无明显差异(P>0.05);CD41、CD61在三组间无显著性差异(P>0.05)。结论血小板CD62p对判断糖尿病病人的血小板活化及血管病变的早期诊断有重要价值。     

2型糖尿病患者血小板CD41、CD61、CD62p的表达及意义

目的研究2型糖尿病患者血小板膜糖蛋白变化与血管病变的关系及其影响因素.方法用流式细胞术测定35例2型糖尿病有血管病变患者、32例2型糖尿病无血管病变患者及31例健康者血小板膜糖蛋白CD41、CD61、CD62p的表达.结果2型糖尿病伴血管病变组CD62p显著高于无血管病变组及健康对照组(P＜0.001),无血管病变组与正常对照组无明显差异(P＞0.05);CD41、CD61在三组间无显著性差异(P＞0.05).结论血小板CD62p对判断糖尿病病人的血小板活化及血管病变的早期诊断有重要价值.     

阿司匹林联合氯吡格雷治疗对急性脑梗死患者的影响

目的:探讨阿司匹林联合氯吡格雷治疗对急性脑梗死患者神经功能及血小板活化的影响。方法:急性脑梗死患者83例随机分为单抗组和双抗组,单抗组行阿司匹林(100 mg/d)单药治疗,双抗组行阿司匹林(100 mg/d)与氯吡格雷(75 mg/d)联合治疗,均治疗2周。于治疗前、治疗1、2周后采用美国国立卫生研究院卒中量表(NIHSS)对2组患者的神经功能缺损程度进行评分并比较,同时使用流式细胞仪术检测并比较2组患者血小板活化指标:血小板上α颗粒膜糖蛋白(CD62p)、溶酶体颗粒膜糖蛋白(CD63)及血小板-单核细胞聚集体(PMA)。结果:治疗1周后,2组的NIHSS评分均显著低于同组治疗前(P0.01);治疗2周后,2组NIHSS评分显著低于同组治疗1周后(P0.01),且双抗组的NIHSS评分显著低于单抗组(P0.01)。治疗1周后,2组的CD62p、CD63、PMA均显著低于同组治疗前(P0.01);治疗2周后,双抗组CD62p、CD63、PMA显著低于同组治疗1周后(P0.01),单抗组CD62p、CD63显著低于同组治疗1周后(P0.01);治疗2周后,双抗组CD62p、CD63、PMA显著低于单抗组(P0.01)。结论:阿司匹林联合氯吡格雷治疗对急性脑梗死治疗效果显著,可有效改善患者神经功能,并可有效抑制血小板活化。     

复方丹参滴丸对自发性高血压大鼠血小板CD62 p和CD63表达的影响

目的：观察复方丹参滴丸对自发性高血压大鼠（SHR）血小板CD62p和CD63表达的影响。方法按所给药物不同,将80只SHR随机分为复方丹参滴丸加阿司匹林组、复方丹参滴丸组、阿司匹林组和生理盐水对照组各20只,各组持续给药4周,于实验12周末采用流式细胞仪检测治疗后SHR血小板CD62p和CD63的表达水平,并对各组结果进行比较。结果第12周末复方丹参滴丸组血小板CD62p、CD63表达水平低于对照组（P＜0.05）,复方丹参滴丸加阿司匹林组CD62p、CD63表达水平较阿司匹林组或复方丹参滴丸组低（P＜0.05）。结论复方丹参滴丸具有抑制血小板CD62p、CD63表达的作用,复方丹参滴丸与阿司匹林合用治疗效果更显著。     

体外循环对血小板功能的影响

目的探讨体外循环(CPB)对血小板计数、血小板膜糖蛋白及血小板聚集功能的影响。方法随机选取15例择期CPB下行心脏手术的病人,在全身肝素化前、肝素化后10min、转流30min、转流结束肝素中和后10min及术后24h分别检测血小板数(PLT)、血小板聚集率(PAGM)及血小板膜糖蛋白(CD41a、CD42a、CD61、CD62p)阳性表达率和平均荧光强度(meanfluorescenceintension,MFI)的变化。结果肝素化后10minPLT及PAGM明显下降,转流结束肝素中和后10min为最低,术后24h渐趋恢复;CD41a、CD42a、CD61的阳性表达率各时段间无明显差异(P>0.05),但CD41a、CD61的MFI明显增强,CD42a的MFI明显减弱,与转流前相比有明显差异(P<0.05、P<0.01);CD62p的阳性表达率及MFI在各时段间均有显著差异(P<0.05、P<0.01)。结论CPB过程中,血小板数量急剧下降,聚集功能受损;血小板膜糖蛋白的表达受明显影响。     

老年高血压肾病血小板活化分子检测的意义

目的 :评估血小板在老年高血压肾病中的活化状态及意义。方法 :利用流式细胞仪检测了 64例老年高血压肾病患者及 2 1例健康自愿者外周血小板的膜糖蛋白CD62P、CD63、CD 41和CD61的表达。结果 :老年高血压肾病组CD62P、CD63的表达较对照组显著增高 (P <0 0 0 1) ,CD41和CD61的表达与对照组比较差异无显著性(P >0 0 5 )。老年高血压肾病组CD62P、CD63的表达组间差异有显著性 (P <0 0 5 ) ,且随血压分级水平升高而递增。结论 :老年高血压肾病时血小板活化增强 ,推测其在老年高血压肾病发生发展中起重要作用。     

Modification of activation-dependent platelet antigens CD62p and CD63 during haemodialysis

In particular, activated platelets are thought to be involved in the pathophysiology of thrombotic occlusions of vessels. In this study, we evaluated activation-dependent changes in platelet antigens during extracorporeal haemodialysis treatment. Flow cytometry was used in combination with monoclonal antibodies that bind to platelet glycoproteins CD62p (GMP-140) and CD63 (GP53). Maximum peaks of mean channel fluorescence intensity (MCFI) were reached after 60 min in 20/26 procedures in CD62p (P < 0.005) and in 15/25 treatments in CD63 (p < 0.002), respectively. An initial peak of CD62p and CD63 fluorescence expression could be detected in 21/25 and 23/25 treatments, respectively (CD62p within 15, CD63 within 30 min), indicating the early onset of activation. The structural antigen CD41a MCFI slightly decreased over time in all treatments, while CD42b expression did not change. From these results we conclude that haemodialysis contributes to platelet activation and secondary hypercoagulability. Analysis of platelet glycoproteins by flow cytometry may provide clinical information on patients at a higher risk for thrombosis and may help in further improvement of haemodialysis equipment.     

Flow cytometric analysis of platelet membrane antigens during and after continuous-flow plateletpheresis

BACKGROUND: The influence, extent, and duration of changes in platelet antigen expression caused by blood-biomaterial interaction in plateletpheresis were assessed. STUDY DESIGN AND METHODS: Twenty-two apheresis donors were studied by using two automated continuous-flow apheresis devices. Blood samples were taken before, during, and for 4 days after extracorporeal circulation. The platelet surface expression of glycoproteins CD41a, CD42b, CD62p, and CD63 was analyzed by flow cytometry. RESULTS: Over the course of plateletpheresis, there was a significant increase in mean channel fluorescence intensity (MCFI) of CD62p, from 25.1 +/− 7.9 (mean +/− SD) to 50.4 +/− 28.9, and of CD63, from 22.3 +/− 6.5 to 33.3 +/− 13.2. There was a significant decrease in CD41a expression as measured by the MCFI, from 1129.8 +/− 125.0 to 1066.6 +/− 102.2, and in CD42b MCFI, from 329.6 +/− 49.4 to 321.4 +/− 52.0. The two apheresis devices showed different platelet activation kinetics, but the overall MCFI of CD62p and CD63 did not significantly diverge after 60 minutes of apheresis. CD62p and CD63 expression as measured by the MCFI returned to preapheresis levels during the follow- up period in 25 and 25 of 44 procedures, respectively, within 24 hours; in 10 and 13 of 44 procedures after 48 hours; in 7 and 3 of 44 procedures after 72 hours; and in 2 and 3 of 44 procedures on Day 5. CONCLUSION: The varying kinetics of expression, as measured by the MCFI, of platelet antigens CD62p, CD63, CD41a, and CD42b during extracorporeal circulation may be useful for biocompatibility testing. Activated platelets continue to circulate in donors for several days after cytapheresis, which suggests that a sufficient interval between apheresis procedures is necessary to avoid the collection of activated platelets.     

Effect of amphotericin B and fluconazole on platelet membrane glycoproteins

BACKGROUND : Fever, chills, and reduced platelet recovery may result when platelets are transfused simultaneously with amphotericin B. Amphotericin B reportedly increases the pitting of membranes in stored platelets. STUDY DESIGN AND METHODS : The effects of amphotericin B and another antifungal agent, fluconazole, on platelet membrane glycoproteins (GP) were examined by the incubation of split aliquots of fresh and stored platelet concentrates (PCs) with these drugs for 3 days in storage bags. To determine the effect of storage, PCs were stored for 5 days, and aliquots removed on Days 1 through 5 were placed in platelet storage bags with 4 micrograms per mL of amphotericin B for 2 to 6 hours. Membrane glycoprotein expression was assessed by flow cytometry with fluorescein isothiocyanate-labeled monoclonal antibodies (MoAbs) directed against the following antigens: GPIb (CD42b), CD63 (an activation protein), P-selectin (CD62), and GPIIb/IIIa (CD41a). RESULTS : Amphotericin B produced a concentration-dependent decrease in the surface binding of CD42b MoAb with no consistent changes in the binding of CD41a, CD63, or CD62 MoAbs after a 3-day exposure. Stored but not fresh PCs showed decreased binding of MoAb CD42b after a 6-hour exposure to amphotericin B (4 micrograms/mL). Fluconazole produced no changes. When the binding of MoAb CD42b to permeabilized platelets was used to measure total platelet content, amphotericin B (4 micrograms/mL) decreased MoAb CD42b binding to a similar degree in fresh and stored platelets. Inhibition of aggregation to ADP and collagen and ADP and epinephrine was seen in stored but not fresh PCs. CONCLUSION : Therapeutic levels of amphotericin B resulted in partial loss of total platelet GPIb in fresh and stored PCs, but decreased surface expression of platelet membrane GPIb only in stored platelets. This difference between fresh and stored platelets may be related to the limited reservoir of GPIb available for redistribution to the membrane in the previously stored PCs and may account for the decreased recovery of transfused platelets observed in some patients receiving amphotericin B.     

Biocompatibility of a new cell separator studied by flow cytometry: analyses of platelet antigens during apheresis and storage

BACKGROUND: Alterations of platelet antigens are known to occur during cytapheresis and storage. These changes have been shown to be dependent on the biomaterials, techniques, and devices used. In this study, the influence of a new cell separator (AMICUS) and storage container (PL-2410) on platelet glycoproteins was analyzed. STUDY DESIGN AND METHODS: During plateletpheresis and storage, the levels of platelet glycoproteins and binding of fibrinogen were determined by flow cytometry. RESULTS: During apheresis, mean channel fluorescence intensity of CD41 a did not change significantly (p = 0.06). A small increase was evident in CD42b mean channel fluorescence intensity, which rose from a baseline level of 178.6 +/- 68.3 to 231.5 +/- 97.9 at the end of the procedure (p<0.05); in CD62p-positive platelets, which increased from 2.0 +/- 0.9 percent to 9.9 +/- 3.9 percent (p<0.05); in CD63-positive platelets, which increased from 1.7 +/- 0.7 percent to 7.9 +/- 2.6 percent (p<0.05); and in the binding of fibrinogen, which increased from 1.9 +/- 0.8 percent positive platelets to 10.5 +/- 2.6 percent (p<0.05). During storage, the mean channel fluorescence intensity of CD41a and CD42b, the percentage of CD62p- and CD63-positive platelets, and the binding of fibrinogen to platelets showed no significant change. CONCLUSION: These studies show that alterations in platelet antigens and platelet activation occur to a small degree during apheresis and storage. These findings demonstrate generally good biocompatibility of this new cell separator.     

Effects of in vitro exposure of alcohol on surface receptor expression of human platelets

Platelet inhibition after moderate alcohol consumption in patients with ischaemic heart disease may contribute to reducing the risk for developing acute coronary syndromes. However, the mechanism by which ethanol affects platelets is not clarified. We sought to determine the in vitro effects of alcohol on the surface expression of human platelet receptors using whole blood flow cytometry. Blood samples from 10 healthy volunteers were incubated for 30 min with 25 and 50 mmol l(-1) of phosphate buffered saline diluted grain ethanol, concentrations often used in in vitro studies. The surface expression of platelet receptors was determined by flow cytometry after fixation with 2% paraformaldehyde using the following monoclonal antibodies: CD 41 (GP IIb/IIIa), CD 42b (GP Ib), CD 62p (P-selectin), CD 51/CD 61 (vitronectin receptor), CD 31 (PECAM-1), CD 107a (LAMP-1), CD 107b (LAMP-2), CD 63 (LIMP, LAMP-3) and CD 151 (PETA-3). Dose-dependent inhibition of GP IIb/IIIa, P-selectin, CD 63 and CD 107a receptor expression was observed in the ethanol-treated whole blood samples. This study for the first time establishes a direct effect of ethanol on selective major platelet receptors. Beneficial cardiovascular properties of moderate alcohol consumption may be explained by ethanol's antiplatelet action.     

Annexin V and platelet antigen expression is not altered during storage of platelet concentrates obtained with the AMICUS cell separator.

During storage of platelet concentrate the so-called "storage lesion" occurs. During this time, platelets loose their morphological and functional capacities that are necessary for proper in vivo efficacy following transfusion. Annexin V represents a marker for apoptosis. In this study, Annexin V and additional antigens were analyzed by flow cytometry. Platelet concentrates were obtained with a new cell separator (AMICUS Separator, Fenwal). Following apheresis, platelet units were stored for an experimentally prolonged time of seven days. Daily aliquots of the platelet-rich plasma were obtained to measure Annexin V and platelet antigens CD62p, CD63, CD41a, CD42b, and the binding of fibrinogen. All analyses were performed using flow cytometry. During storage, no significant changes in mean channel fluorescence intensity (MCFI) of CD41a (P = 0.99) and CD42b (P = 0.29), percentage of CD62p+ and CD63+ platelets (P = 0.23 for CD62p; P = 0.52 for CD63), and the binding of fibrinogen to platelets occurred (P = 0.85). Also, the expression of Annexin V remained constant with no significant change (P = 0.36). This study shows that antigens of platelets, obtained with the AMICUS cell separator are well preserved during storage. Regarding Annexin V, no obvious signs of apoptosis can be detected by flow cytometry. These findings demonstrate the high degree of biocompatibility of the apheresis device and storage container.     

Methylprednisolone pulse therapy in Kawasaki disease

A role of intravenous methylprednisolone pulse (IVMP) therapy has not been established for Kawasaki disease (KD) patients unresponsive to initial intravenous immunoglobulin (IVIG) therapy. We conducted a control study in 22 KD patients unresponsive to initial IVIG to compare IVMP with additional IVIG. IVMP induced faster but temporary resolution of fever and more adverse effects such as bradycardia. For the last four years, 62 KD patients unresponsive to initial IVIG have been treated with additional IVIG, and 17 unresponsive to additional IVIG with IVMP, followed by oral prednisolone. Among of them, coronary artery lesions were detected in two patients, but regressed in 6 months. This protocol may be useful for eradication of coronary artery lesions.     

血小板糖蛋白与高血压、脑梗死关系的临床研究

目的通过测定高血压、脑梗死患者血小板糖蛋白（CD62，CD63，PAC-1）和纤维蛋白原（FIB）指标，旨在为临床治疗高血压、脑梗死提供有用的参考指标。方法采用流式细胞术（FCM），对85例高血压脑梗死患者，42例正常血压脑梗死患者、35例原发性高血压患者血小板糖蛋白CD62P，CD63，PAC-1进行检测。利用Acl-Advanee血凝仪测定了FIB。结果高血压脑梗死组、正常血压脑梗死组与原发性高血压组血小板糖蛋白CD62p、CD63、PAC-1及FIB均显著高于正常对照组（均P〈0．001），且高血压脑梗死组、正常血压脑梗死组血小板糖蛋白CD62p、CD63、PAC．1及FIB均明显高于高血压组（均P〈0．001）。高血压脑梗死组与正常血压脑梗死组血小板糖蛋白CD62p、CD63、PAG-1及FIB均无明显差异。血小板糖蛋白及FIB大梗死灶组〉中梗死灶组〉小梗死灶组，各组之间均有明显差异（P均〈0．01）。血小板糖蛋白CD62P、CD63、PAC-1与FIB呈正相关结论脑梗死、高血压患者均存在血小板活化，脑梗死患者血小板活化与病情及梗死灶大小有关。血小板糖蛋白的检测对高血压患者的病情分析和脑梗死的预防以及早期诊断和治疗具有重要的意义。