P选择素在原发性高血压伴腔隙性脑梗死患者发病机制中的作用

目的 观察血小板活化的标志——P-选择素在原发性高血压伴腔隙性脑梗死患者发病机制中的作用。方法 利用FITC标记的抗CD42a单抗（鼠IgG1)，PE标记的抗CD62P单抗（鼠IgG1），CY标记的抗CD45单抗（鼠IgG1),FITC标记的鼠IgG1同种型对照免疫球蛋白，PE标记的鼠IgG1同种型对照免疫球蛋白，上述单抗分别标记全血中血小板及白细胞的相关抗原；同时利用流式细胞仪，采取全血三色法流式细胞术方法，检测全血血小板上P-选择素阳性表达：全血标本中分别加入3种荧光标记的单克隆抗体FITC-CD42a，PE-62P及CY-CD45，室温避光反应20min，10g/L多聚甲醛固定10min，用PBS洗2次，上机分析。结果 原发性高血压伴腔隙性脑梗死患者血小板P-选择素阳性表达[(23.0&;#177;4.0)%]明显高于原发性高血压无腔隙性脑梗死患者[(16.9&;#177;1.8%)]及正常人组[(5.2&;#177;0.7)%],三者之间相比差异有显著性意义（t=137.4,28.4,P&;lt;0.05).结论 P-选择素在原发性高血压伴腔隙性脑梗死发病中起重要作用，P-选择素与血管渐进损伤有关。     

伴抑郁障碍糖尿病患者的血小板活化功能变化

目的:探讨糖尿病伴抑郁障碍患者的血小板活化功能变化,以及与血管并发症发生之间的关系。方法:根据汉密顿抑郁量表评分将苏州大学附属第一医院内分泌科80例明确诊断的住院糖尿病患者分组,正常对照组为80名体检正常的健康人,应用流式细胞术和酶联免疫分析法检测血小板膜P-选择素表达、纤维蛋白原结合反应、血浆P-选择素水平和血浆血栓烷B2水平的改变并进行比较。结果:伴抑郁障碍的糖尿病患者平均每血小板表达的P-选择素分子数犤(946.71±728.54)个犦、纤维蛋白原结合反应犤(36.71±14.69)%犦、血浆血栓烷B2水平犤(8.56±6.13)nmol/L犦和血浆皮质醇水平犤(11.97±7.15)μg/L犦均高于正常对照组,差异有显著性意义(F=14.85,χ2=10.02,F=3.09,F=11.05,P<0.05～0.01)。抑郁障碍的糖尿病患者的平均每血小板表达的P-选择素分子数、纤维蛋白原结合反应、血浆皮质醇水平和生长激素水平均高于不伴抑郁组,差异有显著性(F=14.85,χ2=10.02,F=11.05,F=2.83,P<0.05～0.01)。结论:伴抑郁障碍的糖尿病患者血小板活化较其他糖尿病患者和正常人群高,可能成为血管并发症的因素之一。     

老年糖尿病患者脑梗死发生与血小板活化和颈动脉粥样硬化的关系

目的:探讨老年2型糖尿病血小板活化和颈动脉粥样硬化病变与脑梗死的关系。方法:108例患者分为2组,包括2型糖尿病76例(糖尿病组),男40例,女36例;糖耐量减退(impairedglucosetolerance,IGT)32例(IGT组),男24例,女8例。采用流式细胞仪测定CD62p和CD41+CD11b,同时使用彩色多普勒显像仪进行颈动脉超声检查。结果:糖尿病组急性脑梗死患者CD62p,CD41+CD11b水平犤(9.2±3.6)%,(98.2±16.5)%犦与未发生脑梗死犤(3.8±1.1)%,(55.6±11.8)%犦比较,差异有显著性意义(t=3.05,2.69,P<0.05);IGT组急性脑梗死与未发生脑梗死患者上述两项指标比较,差异也有显著性意义(t=2.40,2.15,P<0.05);腔隙性脑梗死患者CD62p,CD41+CD11b水平亦高于未发生脑梗死患者(t=2.15～3.05,P<0.05);糖尿病组和IGT组发生急性脑梗死患者CD62p,CD41+CD11b水平与腔隙性脑梗死患者比较,差异有显著性意义(t=2.29～2.79,P<0.05)。老年2型糖尿病组颈动脉有斑块患者发生急性脑梗死(50%)高于发生腔隙性脑梗死(38%),差异有显著性意义(χ2=5.23,P<0.05);颈动脉有斑块者与正常者比较脑梗死及腔隙性脑梗死发生率,差异有显著性意义(χ2=4.12,3.95,P<0.05)。结论:老年2型糖尿病和IGT患者CD62p,CD41+CD11b的测定对缺血性脑卒中的早期诊断和病情监测有一定价值,颈     

老年脑梗死患者血浆神经肽Y、降钙素基因相关肽水平的动态变化

目的:观察老年脑梗死患者神经肽Y、降钙素基因相关肽(calcitoningene-relatedpeptide,CGRP)水平改变及其动态变化,探讨神经肽Y、CGRP参与老年脑梗死的病理生理机制及其临床意义。方法:选择老年脑梗死患者42例,依病情轻重分为轻型12例,中型18例,重型12例。依病灶大小分为大灶组22例,小灶组20例。按病程分为≤24h组19例,1～3d组22例、4～7d组27例、8～15d组21例及>15d组18例。按伴发病积分分为<6分组25例与≥6分组17例。按有无高血压史分为有高血压史组18例与无高血压史组24例。选择30例健康老人查体者作为对照组。用放免法检测血浆神经肽Y与CGRP浓度。结果:神经肽Y的变化:老年脑梗死组神经肽Y水平犤(590.66±305.45)ng/L犦显著高于对照组犤(114.76±65.92)ng/L犦(t=5.2846,P<0.0001),于发病后24h内犤(439.13±237.80)ng/L犦显著升高,1～3d犤(668.54±261.44)ng/L犦达高峰,8～15d犤(528.59±215.67)ng/L犦开始下降,15d犤(372.80±200.96)ng/L犦后仍在较高水平;重型犤(698.96±206.85)ng/L犦与大灶组犤(769.61±296.48)ng/L犦显著高于轻型犤(488.60±201.75)ng/L犦与小灶组犤(483.40±312.31)ng/L犦(P<0.01),发病积分≥6分组犤(631.28±286.86)ng/L犦显著高于<6分组犤(573.35±337.64)ng/L犦(t=2.9687,P<0.     

高血压伴胰岛素抵抗的腔隙性脑梗死患者的代谢特征

目的分析伴胰岛素抵抗的高血压病腔隙性脑梗死患者的代谢特征,为临床治疗提供理论依据。方法1999-6/2001-3在瑞金医院高血压科住院的高血压病患者,经头颅MRI检查确诊为腔隙性脑梗死者中,应用改良的Bergman的减少样本数微小模型方法评价高血压病非糖尿病伴腔梗患者的胰岛素敏感性,分析其胰岛素抵抗伴有的代谢紊乱。结果腰臀比:胰岛素抵抗组(38例)明显大于非胰岛素抵抗组(22例),差别有显著性意义(0.88±0.05vs0.85±0.06,t=2.15,P<0.05);胰岛素抵抗组三酰甘油明显高于非胰岛素抵抗组犤(2.41±0.06)mmol/Lvs(1.48±0.03)mmol/L,t=2.24,P<0.05犦;胰岛素抵抗组胰岛素敏感性和葡萄糖利用率明显低于非胰岛素抵抗组犤(4.0±0.6)min/mU·Lvs(4.8±0.6)min/mU·L,t=2.18,P<0.05;(2.12±0.17)/minvs(2.61±0.18)/min,t=2.24,P<0.05犦。结论高血压病非糖尿病伴腔隙性脑梗死的胰岛素抵抗患者同时伴有腹部肥胖和高三酰甘油。     

运动疗法对充血性心力衰竭患者凝血功能的影响

目的:研究运动治疗对充血性心力衰竭患者凝血功能的影响。方法:将57例充血性心力衰竭患者,分为运动组(28例)和对照组(29例),运动组进行运动康复治疗七八周;而对照组不进行运动康复治疗,观察实验前后的临床表现、血浆血管性假性血友病因子(vonWillebrandfactor,vWF)、P-选择素和D-二聚体水平变化。结果:两组实验前血浆vWF、P-选择素和D-二聚体水平无显著差异,运动组血浆vWF犤(138.96±33.84)%犦、P-选择素犤(19.02±3.97)g/L犦和D-二聚体犤(0.60±0.14)mg/L犦水平均较实验前犤(176.88±34.87)%,(24.46±4.32)g/L,(0.78±0.13)mg/L犦下降(t=4.12～4.98,P<0.05);对照组实验后血浆vWF、P-选择素和D-二聚体水平也较实验前下降(t=2.08～2.38,P<0.05)。实验后运动组血浆vWF、P-选择素和D-二聚体水平较对照组下降明显(t=2.12～2.41,P<0.05)。结论:运动康复可改善充血性心力衰竭患者的凝血功能,可能在一定程度上减少充血性心力衰竭患者血栓事件的发生和发展。     

原发性高血压患者血浆活化血小板表达的研究

目的：探讨高血压病患者血小板活性因子指标CD62P（P-选择素）、CD63的变化。方法：取158例原发性高血压患者血浆，用流式细胞仪进行检测CD62P（P-选择素）、CD63，自动生化仪测定肾功能、放射免疫法测定血、尿微球蛋白。结果：高血压组CD62P（P-选择素）、CD63均明显高于正常对照组，且随高血压病患者血压分级水平的增高而增高。结论：高血压病患者分级水平的增高与血小板活化指标CD62P（P-选择素）和CD63过度表达有一定的关系，表明其可能参与高血压的发生、发展。     

急性脑梗死患者外周血血小板表面P-选择素阳性表达率变化及药物干预影响

目的 探讨血小板活化因子P-选择素在急性脑血管疾病患者中的改变及应用奥扎格雷钠治疗后的变化。方法采用流式细胞仪测定急性脑梗死患者及正常对照组外周血血小板表面P-选择素阳性表达情况，对比观察治疗前后P-选择素的变化。结果急性脑梗死患者外周血血小板表面P-选择素阳性表达率显著高于正常对照组（P〈0．01）；应用奥扎格雷钠治疗后的外周血血小板表面P-选择素阳性表达率明显低于正常对照组（P〈0．01）。结论血小板活化在急性脑梗死的发生、发展中起重要作用；P-选择素可作为选择药物治疗的一项可靠指标，奥扎格雷钠可减少血小板活化。     

灯盏花素合用脑复素治疗原发性高血压并发腔隙性脑梗死

目的:探讨治疗原发性高血压并发腔隙性脑梗死的有效方法。方法:将原发性高血压并发腔隙性脑梗死62例随机分为2组,治疗组30例,对照组32例,治疗组给予灯盏花素合脑复素治疗,对照组给予丹参合胞二磷胆碱常规治疗。结果:治疗组有效率为86.7%,对照组为71.9%,2组比较有显著差异(P<0.05)。结论:灯盏花素合用脑复素治疗原发性高血压合并腔隙性脑梗死疗效满意。     

急性脑血管病外周血血小板表面P-选择素阳性表达率变化及药物干预影响

目的为了研究血小板活化因子P-选择素(CD62P)在急性脑血管病患者中的改变及其用奥扎格雷钠治疗后变化。方法采用流式细胞仪测定急性脑梗死患者及健康对照组外周血血小板表面P-选择素阳性表达率,治疗前后测P-选择素作对比观察。急性脑出血患者入院后测外周血P-选择素与健康对照组作对比。结果1·急性脑梗死患者外周血血小板表面P-选择素阳性表达率显著高于对照组(P<0·001)。2·用奥扎格雷钠治疗后的外周血血小板表面P-选择素阳性表达率明显低于非奥扎格雷钠组(P<0·01)。结论血小板活化在急性出血及脑梗死的发生、发展中起重要作用。P-选择素可以做为选择药物治疗的一项可靠指标。奥扎格雷钠具有减少血小板活化。     

Platelet activation in acute pulmonary embolism

BACKGROUND: Platelet activation is implicated in thrombotic disorders, but has not been described in acute clinical pulmonary embolism (PE). Objectives: To investigate the natural history of platelet activation in PE and associated markers of inflammation, thrombosis and cardiac dysfunction. METHODS: Thirty-five consecutive patients (age 62 +/-17 years) with acute PE were prospectively enrolled and followed for 6 months. Platelet activation was assessed by flow cytometry [measuring expression of platelet P-selectin, conformational activation of glycoprotein IIb/IIIa complex (PAC-1) and formation of platelet-leukocyte complexes] and by plasma soluble P-selectin. Platelet activation, right ventricular (RV) function (assessed as RV ejection area by transthoracic echocardiography), D-dimer and high-sensitivity C-reactive protein (hs-CRP) were measured at presentation and repeated over 6 months follow-up. RESULTS: Soluble P-selectin (56 +/-19 ng mL(-1), anovaP < 0.0001) and PAC-1 (1.5 +/- 1.8%, anovaP = 0.005) were mildly but significantly increased in patients with acute PE relative to healthy young men (soluble P-selectin 33 +/- 13 ng mL(-1), P < 0.001; PAC-1 binding 0.5 +/- 0.6%, P < 0.01) and age-matched controls (soluble P-selectin 31 +/- 9 ng mL(-1), P < 0.001; PAC-1 binding 0.4 +/-0.4%, P < 0.05). Platelet P-selectin expression and platelet-leukocyte complexes were not increased during acute PE. Echocardiographic RV ejection area correlated inversely with soluble P-selectin (r = -0.47, P = 0.007) and positively with platelet P-selectin (r = 0.49, P = 0.0007), suggesting P-selectin is shed from activated platelets in proportion to the severity of RV dysfunction. Elevated soluble P-selectin, D-dimer and hs-CRP demonstrated a time-dependent return to normal during 6 months follow-up. CONCLUSION: Platelet activation is evident after acute PE. Platelet activation correlates with the severity of RV dysfunction, and can persist for several months after acute PE.     

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。     

丁胺卡那霉素对血小板聚集和凝血功能的抑制作用

目的 观察丁胺卡那霉素对血小板聚集和凝血功能试验的抑制作用,探讨其对止血功能的影响及相关作用机制.方法 将不同浓度丁胺卡那霉素分别与献血者富血小板血浆和乏血小板血浆作用,分为4组:0 mg/L组、30 mg/L组、91 mg/L组和910 mg/L组.以血小板聚集分析仪检测ADP诱导的血小板最大聚集率,以流式细胞仪检测活化血小板P-选择素、GPⅡb/Ⅲa及Fg-R表达水平,以血液凝固分析仪测定PT、APTT、TT及Fg水平.以前述4种浓度丁胺卡那霉素以及62.5 U/ml肝素钠和109 mmoL/L柠檬酸钠分别与新鲜全血作用,测定CT及血浆Ca2+浓度.分别检测10例急性下呼吸道感染患者在丁胺卡那霉素常规剂量治疗前和治疗后30 min ADP诱导的血小板最大聚集率、P-选择素、GPⅡb/Ⅲa及Fg-R表达水平,并测定PT、APTT、CT和血浆Ca2+浓度.结果 30 mg/L组血小板最大聚集率、P-选择素和Fg-R分别为(65.8±3.9)%、(9.2±1.0)%和(12.6±1.7)%,显著低于0 mg/L组的(88.0±4.6)%、(16.1±1.3)%和(31.0±2.5)%,差异均有统计学意义(t值分别为9.442、8.432和9.993,P均＜0.01);30 mg/L组APTT(80.5±6.8)s和CT(857±66)s明显高于0 mg/L组的(33.0±3.6)s和(447±35)s,差异均有统计学意义(t值分别为11.312和13.211,P均＜0.01);丁胺卡那霉素浓度与血小板最大聚集率呈显著负相关,与聚集的抑制率呈显著正相关,与APTT呈显著正相关[r值分别为-0.832、0.939和(＞0.870),P均＜0.05];30 mg/L组,91 me/L组和910 mg/L组呈剂量依赖性抑制P-选择素和Fg-R表达及使CT增加[F组间=21.44、26.24和(＞29.81),P均＜0.01].0 mg/L组、30mg/L组、91 mg/L组和910 mg/L组PT值分别为(14.7±1.9)s、(15.2±1.7)s、(15.6±1.5)s、(22.1±2.1)s,差异有统计学意义(F=8.21,P＜0.05),而GPⅡb/Ⅲa、TT、Fg以及血浆Ca2+浓度在4组间差异无统计学意义(P均＞0.05).丁胺卡那霉素治疗后患者血小板最大聚集率(51.6±10.1)%、P-选择素(6.8±1.8)%和Fg-R(20.1±5.8)%明显低于治疗前的(66.8±11.4)%、(10.9±3.1)%和(28.5 ±7.4)%,APTT(49.8±5.9)s和CT值(660±59)s则明显高于治疗前的(26.9±3.8)s和(410±45)s,差异均有统计学意义(t值分别为5.456、8.875、7.423、10.012和11.322,P均＜0.01).治疗前、后GPⅡb/Ⅲa、PT和Ca2+浓度变化无统计学意义(P＞0.05).结论 丁胺卡那霉素通过抑制血小板纤维蛋白原受体活化和释放反应途径抑制血小板聚集,可能通过抑制内源凝血系统因子途径而抑制凝血功能,从而对止血功能有抑制作用;应用丁胺卡那霉素抗感染治疗可能有发生出血的危险.     

Accumulation of tissue factor into developing thrombi in vivo is dependent upon microparticle P-selectin glycoprotein ligand 1 and platelet P-selectin

Using a laser-induced endothelial injury model, we examined thrombus formation in the microcirculation of wild-type and genetically altered mice by real-time in vivo microscopy to analyze this complex physiologic process in a system that includes the vessel wall, the presence of flowing blood, and the absence of anticoagulants. We observe P-selectin expression, tissue factor accumulation, and fibrin generation after platelet localization in the developing thrombus in arterioles of wild-type mice. However, mice lacking P-selectin glycoprotein ligand 1 (PSGL-1) or P-selectin, or wild-type mice infused with blocking P-selectin antibodies, developed platelet thrombi containing minimal tissue factor and fibrin. To explore the delivery of tissue factor into a developing thrombus, we identified monocyte-derived microparticles in human platelet-poor plasma that express tissue factor, PSGL-1, and CD14. Fluorescently labeled mouse microparticles infused into a recipient mouse localized within the developing thrombus, indicating that one pathway for the initiation of blood coagulation in vivo involves the accumulation of tissue factor- and PSGL-1-containing microparticles in the platelet thrombus expressing P-selectin. These monocyte-derived microparticles bind to activated platelets in an interaction mediated by platelet P-selectin and microparticle PSGL-1. We propose that PSGL-1 plays a role in blood coagulation in addition to its known role in leukocyte trafficking.     

冠心病患者血小板膜糖蛋白CD_(62)P表达和血粘度的临床研究

目的　探讨血小板膜糖蛋白CD62 P表达及血粘度与冠心病的关系。方法　利用流式细胞仪和单克隆抗体技术测定 82例冠心病患者 (稳定型心绞痛 2 6例A组 ;不稳定型心绞痛 2 8例B组 ;急性心肌梗死2 8例C组和 2 8例健康人D组 )的CD62 P活性及血液流变学指标。结果　A组、B组、C组和D组的CD62 P阳性表达率分别为 3 6 2± 1 2 4 %、5 2 6± 1 4 9%、6 0 8± 1 75 %和 2 4 6± 0 81%。A组、B组和C组与健康人D组比较 ,差异均有显著意义 (P值均 <0 0 5 )。以上各组的血浆粘度分别为 1 31± 0 30、1 6 4± 0 2 2、2 19±0 35及 1 30± 0 18。B组、C组与对照组相比差异均有显著意义 (P值均 <0 0 5 ) ,CD62 P与血浆粘度和纤维蛋白原浓度呈正相关 (r =0 872 ,P <0 0 5 ;r =0 793,P <0 0 5 )。结论　CD62 P及血浆粘度均可能成为判断冠心病患者病情进展程度的良好指标     

脑梗死患者活化血小板的检测及其临床意义

目的探讨脑梗死患者全血血小板膜糖蛋白Ⅱb/Ⅲa(PAC-1)、血小板颗粒膜糖蛋白140(CD62P)的变化及其临床意义。方法用全血流式细胞术测定76例脑梗死患者急性期和恢复期、30例高血压病患者和28例糖尿病患者外周血中血小板PAC-1、CD62P的表达水平,并与20例健康对照者比较。结果脑梗死组急性期、恢复期、高血压病组、糖尿病组PAC-1、CD62P的表达均明显高于正常对照组(均P<0.01)。脑梗死患者急性期PAC-1、CD62P的表达大梗死灶组>中梗死灶组>小梗死灶组,各组之间比较有显著性差异(P<0.01~0.05)。脑梗死患者急性期PAC-1、CD62P的表达重型>中型>轻型,各组之间比较有显著性差异(P<0.01~0.05)。结论脑梗死急性期、恢复期、高血压病患者、糖尿病患者均存在血小板活化;脑梗死病人血小板活化与病情及梗死灶大小有关,PAC-1、CD62P可作为判断病情的指标。     

糖尿病患者血小板膜糖蛋白的改变及其临床意义

目的观察Ⅱ型糖尿病患者血小板膜糖蛋白的变化及其临床意义。方法应用流式细胞术（ＦＣＭ）观察７０例Ⅱ型糖尿病患者及２８名正常人血小板膜上ＧＰⅠｂ、Ⅱｂ、Ⅲａ及Ｐ选择素的表达。以免疫放射法（ＩＲＭＡ）测定血小板膜Ｐ选择素分子数,并以酶联免疫法（ＥＬＩＳＡ）对血浆中的Ｐ选择素进行检测。结果ＦＣＭ测得的血小板膜上Ｐ选择素表达阳性率,糖尿病伴血管病患者为（１０．２８±４．８６）％,明显高于正常人的（３．７５±１．８３）％（Ｐ＜０．０１）及无血管病变者的（４．２５±２．２９）％（Ｐ＜００１）。用ＩＲＭＡ法测得的结果与ＦＣＭ类似。血浆中Ｐ选择素浓度伴血管病变组（６．１８±３．２０）μｇ／Ｌ亦高于正常对照组的（３．４３±１．５６）μｇ／Ｌ（Ｐ＜０．０５）及无血管病变组的（３．７１±２．１２）μｇ／Ｌ（Ｐ＜００５）。相反,血小板上ＧＰⅠｂ的表达在糖尿病伴血管病变组为（８６．３５±１１．００）％明显低于正常对照组的（９２６３±７．５５）％（Ｐ＜０．０５）。而血小板ＧＰⅡｂ、Ⅲａ的表达无改变。结论有血管病变的Ⅱ型糖尿患者血小板膜上和血浆中Ｐ选择素升高,而ＧＰⅠｂ降低。     

Platelet membrane glycoproteins and microvesicles in blood from postoperative salvage: a study in cardiac bypass patients

BACKGROUND: Transfusion of blood collected by intraoperative and postoperative salvage systems has been linked to the development of thrombocytopenia and disseminated intravascular coagulation. Although functional defects have been reported in platelets from unwashed salvaged blood, platelet membrane glycoprotein (GP) composition, a potentially important determinant of function and survival, has not been studied. STUDY DESIGN AND METHODS: Platelets from 22 patients whose blood was salvaged at the completion of surgery were analyzed and compared to platelets obtained from the venous blood from the same patient. Platelet membranes were stained with fluorescein isothiocyanate-conjugated CD41a monoclonal antibody (anti-GPIIb/IIIa) to identify platelets, a phycoerythrin-conjugated monoclonal antibody, CD62 (anti-P-selectin) to identify activated platelets, and CD42b (anti- GPIb) or anti-GPIb/IX to assess GPIb. Samples were analyzed with a flow cytometer using software. RESULTS: Platelets obtained from salvaged blood demonstrated lower GPIb expression (CD42b and GPIb/IX monoclonal antibody binding), higher P-selectin expression, and greater numbers of platelet-derived microvesicles. CONCLUSION: The clinical significance of transfusing blood containing activated platelets and microvesicles merits investigation.     

Effects of platelet binding on whole blood flow cytometry assays of monocyte and neutrophil procoagulant activity

BACKGROUND: Monocytes and neutrophils form heterotypic aggregates with platelets initially via engagement of platelet surface P-selectin with leukocyte surface P-selectin glycoprotein ligand-1 (PSGL-1). The resultant intracellular signaling causes the leukocyte surface expression of tissue factor and activation of leukocyte surface Mac-1 (integrin alphaMbeta2, CD11b/CD18). The activation-dependent conformational change in monocyte surface Mac-1 results in the binding of coagulation factor Xa (FXa) and/or fibrinogen to Mac-1. The aim of this study was to develop whole blood flow cytometry assays of these procoagulant activities and to investigate the effects of platelet binding to monocytes and neutrophils. METHODS: Citrate or D-Phe-Pro-Arg-chloromethylketone (PPACK) anticoagulated whole blood was incubated with monoclonal antibodies against CD14 (PECy5), CD42a (PE), FITC-conjugated test antibody and an agonist, and then fixed with FACS lyse. Appropriate isotype negative controls were prepared in parallel. A BD FACSCalibur was used to analyze monocytes and neutrophils, which were identified based on CD14 fluorescence, forward and 90 degrees light scatter. These populations were further gated into CD42a-positive (platelet-bound) and CD42a-negative (platelet-free). Geometric mean fluorescence and per cent positive data were collected for each subpopulation to measure the binding of test antibodies directed at CD42a, tissue factor, coagulation FXa, bound fibrinogen, activated Mac-1, and CD11b. Compensation controls were prepared on six normal donors prior to the study and these settings were used throughout the 10 donor study. Negative controls verified the lack of cross talk, particularly in the quantified FITC and PE parameters. RESULTS: The physiologic agonists collagen and ADP increased monocyte-platelet and neutrophil-platelet aggregates and increased leukocyte surface Mac-1/CD11b and surface-bound tissue factor, FXa and fibrinogen. Whereas the increases in Mac-1/CD11b were mainly independent of leukocyte-platelet binding, the increases in surface-bound tissue factor, FXa and fibrinogen were mainly dependent on leukocyte-platelet binding. CONCLUSIONS: (i) We have developed novel whole blood flow cytometry assays to measure bound tissue factor, coagulation FXa, fibrinogen, activated Mac-1 and CD11b on the surface of monocytes and neutrophils, allowing independent analysis of monocytes and neutrophils with and without surface-adherent platelets. (ii) The monocyte and neutrophil surface binding of tissue factor, FXa and fibrinogen is mainly dependent on platelet adherence to monocytes and neutrophils, whereas the monocyte and neutrophil surface expression of CD11b and activated Mac-1 is mainly independent of platelet adherence to monocytes and neutrophils.     

耳折征患者颈动脉内膜中层厚度和斑块发生率的临床观察

目的 分析双侧耳折征(DELC)患者的颈动脉内膜中层厚度(IMT)、颈动脉斑块发生率的情况。方法 选取50岁以上患者110例,其中DELC组60例,对照组50例。所有患者测量体质量、身高,记录吸烟史及高血压病、2型糖尿病、脑梗死、心肌梗死病史,应用彩色多普勒超声检查颈动脉结构。对比两组年龄、男性比例、体质量指数(BMI)、吸烟率、患病率(高血压病、2型糖尿病、脑梗死、心肌梗死)、颈动脉IMT、斑块发生率。结果 与对照组相比,DELC组BMI、吸烟者无显著增加,但年龄更大[(72.6±8.4)岁比(66.1±14.7)岁,P0.05],高血压病(75.0%比56.0%,P0.05)、2型糖尿病(45.0%比26.0%,P0.05)、陈旧性脑梗死(43.3%比22.0%,P0.05)、陈旧性心肌梗死(36.7%比16.0%,P0.05)患病率更高。与对照组相比,DELC组颈动脉IMT明显增厚[(1.21±0.27)mm比(0.84±0.21)mm,P0.05)],斑块发生率明显增加(85%比62%,P0.05)。结论 对于DELC患者,应加强动脉粥样硬化及其危险因素的筛查。