THE IMPAIRMENT OF PLATELET FUNCTION IN FIBRINOLYSIS AND PRESERVING EFFECT OF APROTININ

Platelet adhesion depends on the platelet membrane glycoprotein Ib (GPIb) and plasma von Willebrand Factor (vWF), which can be reflected by ristocetin-induced aggregation. Here we report damage effect of fibrinolysis and preserving effect of aprotinin on platelet function. Addition of 40 U/ml urokinase and 0.3 U/ml plasmin to PRP or washed platelets made the ristocetin-induced aggregation decline to 31.6% and 38.5% of control value respectively. The extent of declining was positively correlated with the concentration of urokinase and plasmin. Meanwhile, the platelet GPIb decreased to 76.4% of control value. The results showed that the fibrinolysis impaired the platelet function and this effect may be associated with the hydrolysis of GPIb. Further research found that by adding the same dose of urokinase or plasmin to aprotinin-pretreated PRP or washed platelets, the aggregation did not change statistically and decrement of GPIb is much less marked. We concluded that the aprotinin could relieve the platelet dsfunction effectively by its inhibitory effect on fibrinolytic activity.     

造血干细胞移植预处理对血小板膜糖蛋白表达和聚集功能的影响

目的研究造血干细胞移植预处理对血小板膜糖蛋白表达和血小板聚集功能的影响。方法应用血小板聚集仪检测29例造血干细胞移植患者预处理前后ADP诱导的血小板聚集功能变化,同时用流式细胞仪观察血小板膜糖蛋白（GP）Ⅰb、Ⅱb、Ⅲa和CD62P表达水平的改变。20名健康人为正常对照。结果移植患者预处理前血小板对ADP的最大聚集率为（56.73±20.38）%,和正常人的（57.66±12）%相比,无明显差异（P〉0.05）;预处理后血小板对ADP的最大聚集率降至（31.38±25.81）%,与预处理前比较,差异显著（P〈0.01）。GPⅡb、GPⅢa与CD62P在预处理后较正常组及预处理前明显升高（P〈0.05）,GPⅠb变化不明显。预处理过程中血凝常规无明显改变。结论造血干细胞移植预处理会引起血小板的活化,导致血小板聚集功能与血小板膜GP的改变,这可能是导致出血或血栓等并发症的原因之一。     

Clinical Observation on Effect of Xiaoyu Zhixue Tablet on 104 Patients with ldiopathic Multifocal Bleeding and Pletelet Aggregation Defect

SupportedbythegrantfromtheNationalNaturalScienceFoundationofChina(No.39370322)ClinicalcurativeeffectofselfformulatedXiaoyuZhixueTablet(XZT,atime-honoredpreparationofChineseherbalmedicine)on104patientswithidiopathicmultifocalbleedingandplateletaggregationdefect(IMBPAD)wasobservedintherecent3yearsbytheauthors,theeffectwassatisfactory.METHODSClinicalMaterialsAllthepatientswereoutpatientsofthespecificclinicoftheauthors'hospital.The104patientsintheTCMgroupwere31menand73women,withtheirag…     

腺苷二磷酸在凝血酶信号传递过程中的作用

目的比较腺苷二磷酸（ADP）在两类凝血酶受体活化过程中对血小板聚集率以及血小板膜表面活化标记物表达的影响,探讨ADP在凝血酶信号传递机制中的作用。方法分别以凝血酶受体（PAR）活化肽PAR1-AP与PAR4-AP诱导血小板活化,观测腺苷三磷酸双磷酸酶（Apyrase,ADP抑制剂）ⅤⅡ作用过程中血小板聚集以及血小板膜表面糖蛋白（GP）Ib与P-选择素的改变。结果两类凝血酶受体都能诱导血小板活化,产生完全的聚集波,血小板膜GPIb则呈现先进行性减少后逐渐回升的可逆性变化,P-选择素水平持续升高。ApyraseⅤⅡ作用后,PAR1-AP诱导的血小板聚集反应受到部分抑制,而PAR4-AP诱导的血小板聚集反应未有明显改变。ApyraseVⅡ加速PAR1途径GPIb回复细胞表面,对PAR4途径中GPIb的改变则没有显著影响。两种活化途径中P-选择素的表达都不受ApyraseVⅡ的影响。结论ADP在凝血酶信号传递过程中发挥重要作用,其中PAR1途径与ADP参与密切相关．     

人血小板膜糖蛋白GPIbα（H1-V289）的原核表达及其生物活性研究

目的获得重组的血小板膜糖蛋白GPIbα的N端片段（1-289氨基酸）,进一步研究其在血栓与止血过程中的生物功能。方法利用质粒pCMV3（编码GPIbαH1-V289）设计引物,构建pQE30-GPIbα（H1-V289）表达载体,转化大肠杆菌M15,IPTG诱导表达蛋白。用Ni-NTA琼脂糖层析柱不同pH值梯度淋洗法纯化蛋白,SDS-PAGE和Western blot鉴定纯化产品纯度和免疫学活性,并观察重组蛋白对瑞斯托霉素和ADP诱导血小板聚集的影响。结果成功获得纯度较高的重组GPIbα（H1-V289）蛋白,浓度为0.6 mg/ml。Western blot检测结果显示,抗GPIbα单抗SZ2能在34 kd区域显示条带。而且纯品能有效抑制瑞斯托霉素诱导的血小板聚集,对ADP诱导的反应无抑制作用。结论重组蛋白具有较好的免疫原性和生物活性,并可以大量获得,为开发抗血栓药物奠定了基础。     

氨甲环酸与抑肽酶对血液保护功能的对比研究

目的　比较氨甲环酸(TA)与抑肽酶(AP)对体外循环术中血液保护功能的影响,探讨其机制。方法　选取体外循环心脏手术病人 82例,随机分为氨甲环酸用药组、抑肽酶用药组和对照组。TA组总量 1. 5g,于麻醉诱导后切皮前及体外循环开始后两个时间点各给半量,一次性静脉推注;AP组 500万KIU同TA组两个时间点半量分次持续滴注给药,对照组不用药。于术前、后分别测定血小板数量与功能、平均血小板体积、凝血三项、D-二聚体,记录各组术中出血量、术后 12h、24h纵隔心包引流量及术后输血量。结果　AP组与TA组较空白对照组在术前后血小板数量功能变化、D-dimer值及术后引流量、输血量这些方面均有显著性差异 (P<0. 05),而AP组与TA组在此方面的数值差异均无统计学意义。结论　本实验证明了氨甲环酸在体外循环中作用近似抑肽酶,除明显抑制术中体内纤溶系统激活外,在减少术后血小板数目和功能的减低方面发挥了作用。另外两药均可明显减少术后的引流量、全血及成分血的输入量。     

Platelet Functions in Cardiopulmonary Bypass Surgery

Background

Haemorrhage after Cardio Pulmonary Bypass (CPB) Surgery is a well recognised complication that leads to significant morbidity and mortality. The incidence varies between 5-25% depending upon the clinical situation. Several factors are implicated as causative but none have been precisely proved.

Methods

Our study was an attempt to evaluate the haemostatic defect with particular reference to platelet function abnormalities during cardio pulmonary bypass surgery, in order to reduce the morbidity and mortality associated with post CPB haemorrhage. Flow cytometric evaluation of different platelet glycoproteins like GPIb/IX, GPIIb/IIIa and GMP-140 was done.

Results

The marker expression showed deregulation during surgery which returned to base after bypass was terminated. In contrast, the cases with bleeding showed significant variation. P-Selectin (GMP 140) expression decreased progressively till 3^rd post-operative day showing lack of activation of platelets in cases of severe bleeding.

Conclusion

Longer duration of CPB initiates plasmin generation through heparin, which raises the PAI-1-tPA complex and thereby down regulating the functions of platelets. This suggests a link between duration of CPB, bleeding, platelet dysfunction and fibrinolysis. Hence serial estimations of the levels of GMP-140 and tPA can predict severe bleeding.Key Words: CardioPulmonary Bypass, Platelet dysfunction, flowcytometry, platelet glycoproteins, haemorrhage     

体外循环中血浆神经肽Y变化及抑肽酶的影响

目的 观察体外循环（CPB）期间血浆神经肽Y（NPY）含量的变化，并探讨抑肽酶对其影响。方法 30例CPB心内直视手术患者随机分为2组，对照组10例和小剂量抑肽酶组20例。分别于术前空腹，CPB开始后15min及30min、术后3h、术后24h取静脉血。检测血浆NPY值和血小板计数（PC）。结果 两组患者NPY值于CPB开始后升高，PC于CPB开始后降低；NPY、PC于术后开始逐步恢复但未至正常基础值。对照组CPB期间血浆NPY释放增加与血小板计数下降呈显著负相关（r=-0．648，P〈0．05）。实验组NPY在CPB期间升高幅度低于对照组（P〈0．05），PC的降低幅度低于对照组（P〈0．05）。结论 CPB期间使用抑肽酶能有效地保护血小板、减少血小板数量的下降，抑制血浆NPY的释放，从而有利于循环功能的稳定，减少术后出血。     

尼群地平对体外循环中血小板数量及粘附,聚集功能的影响

４０例先天性心脏病患儿按随机配对原则分为对照组（Ｃ组）与实验组（Ｎ组）。Ｎ组术前７ｄ口服尼群地平０．５ｍｇ·ｋｇ－１／次，１次／８ｈ，最后１次用药于术前２ｈ。测定围体外循环期血小板计数、瑞斯托霉素及ＡＤＰ诱导的最大凝集率。结果表明，Ｎ组血小板计数、瑞斯托霉素诱导的最大凝集率在转流中、后增高于Ｃ组（Ｐ＜０．０５～０．０１），ＡＤＰ诱导的最大聚集率在转流前、中低于Ｃ组，转流后高于Ｃ组（Ｐ＜０．０５～０．０１），术后纵隔引流量明显减少。说明尼群地平能有效地抑制体外循环中血小板活化，保护血小板的数量与功能     

乌司他丁和抑肽酶对心血管手术患者血液的保护作用

目的观察乌司他丁和抑肽酶对体外循环(CPB)心脏手术的血液保护功能。方法选择90例风湿性心脏病瓣膜置换术的患者,随机分成3组。乌司他丁组(30例):给予乌司他丁12000U/kg,于麻醉诱导后至CPB开始前缓慢静注半量,另半量加入预充液中随转机进入体内;抑肽酶组(30例):于CPB中一次性给予抑肽酶5000000U;对照组(30例):给予等量生理盐水。结果与对照组相比,乌司他丁组术后24h胸腔积液量减少33.3%(P<0.05),抑肽酶组术后24h胸腔积液量减少39.8%(P<0.05),乌司他丁组和抑肽酶组相比差异无统计学意义。抑肽酶组有2例发生严重过敏。结论乌司他丁和抑肽酶均具有维持纤溶活性稳定,保护血小板功能,从而减少术中及术后出血量。     

小剂量抑肽酶在不同病种体外循环手术的应用研究

目的 选择房缺、室缺修补术患者与心脏瓣膜转换术患者,应用小剂量抑肽毒,观察其减少术后失血量的效果并加以比较,探讨抑肽酶的作用机制。方法 ４０例房缺、室缺修补术患者分为对照组（Ａ１ ｎ＝２０）,小剂量抑肽酶组（Ａ２ ｎ＝２０）,３８例瓣膜置换术患者分为对照组（Ｂ１ ｎ＝２０）,小剂量抑肽酶组（Ｂ２ ｎ＝１８）,于麻醉后（Ｔ１０、转机后１０ｍｉｎ（Ｔ２）、鱼精蛋白拮抗后１０ｍｉｎ（Ｔ３）、体外循环后２ｈ（Ｔ４）各时间上测定ＦＤＰ（纤维蛋白降解产物）、Ｆｉｂ（纤维蛋白）、ＡＣＴ（全血激活凝固时间）、血小板计数。结果 ＦＤＰ、Ｆｉｂ在Ｔ２、Ｔ３、Ｔ４时Ａ２明显优于Ａ１（Ｐ〈０．０１）,在Ｔ２时Ｂ２与Ｂ１比较有显著性差异（Ｐ〈０．０５）,２４ｈ渗血量Ｂ２明显多于Ａ２（Ｐ〈０．０１）。结论 小剂量抑肽酶在减少术后渗血量方面     

体外循环期间抑肽酶不同给药方式对血小板保护的临床研究

目的：探讨抑肽酶不同给药方式对体外循环（CPB）期间血小板功能的保护效果。方法：选择心脏瓣膜置换术患者20例，在CPB围手术期给予抑肽酶5-8万U/kg，按给药方式不同随机分为2组，对照组在预充液中一次性给药，试验组于麻醉诱导后、机器预充液中、停机后分别给药1/3量。分别于肝素化后10min、CPB30min、主动脉开放时、CPB结束时、CPB结束后2h几个时点，定量检测α-颗粒蛋白（GMP-140）、血栓烷B2(TXB2)、11-去氢-血栓烷B2(DH-TXB2)、血小板计数及术后第一天的胸腔积液量、手术过程的输血量等指标，运用SPSS软件包进行统计学分析。结果：肝素化后10min、CPB结束时、CPB结束后2h，对照组血小板计数较试验组明显减少；GMP-140、TXB2、DH-TXB2在主动脉开放前两组无差异，开放后至术后2h对照组明显高于试验组，术后第一天的出血量及手术的总输血量也明显少于对照组。结论：CPB围术期持续给予抑肽酶对血小板功能的保护优于自机器预充液中一次性给药。     

Protective Effect of Heparin-coated Circuits on the Platelets during Cardiopulmonary Bypass

Duringcardiopulmonarybypass (CPB ) ,me chanicalstressaswellasthecontactofbloodwiththesurfaceoftheCPBcircuitleadstoplateletactivation ,abnormalitiesinplateletfunctionandthrombocy topenia ,whichresultinpostoperativehemostaticdys function .Theaimofthisstudywastoevaluatewhethertheuseofheparin coatedcircuitscouldre duceplateletactivation ,protectplateletsandimprovethebiocompatibilityofCPBsystems .1　MATERIALSANDMETHODS1 1　PatientsandTreatmentFromOctober 2 0 0 0toApril 2 0 0 1,2 3patien…     

融合蛋白TAP-SSL5对人血小板功能的影响

目的探讨抗炎抗凝双效融合蛋白TAP-SSL5对人血小板功能的影响。方法采用健康人富含血小板血浆以凝胶过滤法分离得到人血小板。流式细胞仪检测融合蛋白TAP-SSL5或小鼠抗人CD42b(glycoprotein Ibα,GPIbα)单克隆抗体(HIP1)与血小板的结合情况;以人血小板表面P-选择素的表达及PAC-1的结合反映血小板的激活程度;以全血电阻法定量分析TAP-SSL5对人血小板聚集功能的影响;为评价TAP-SSL5的出血风险,进一步观察了TAP-SSL5对小鼠尾部出血时间的影响。结果融合蛋白TAP-SSL5可与血小板结合,并竞争性抑制HIP1与血小板的结合,提示TAP-SSL5可与血小板表面的GPIbα结合,进而抑制GPIbα与vWF的相互作用。但高浓度的TAP-SSL5(终浓度30 mg/L)也能激活血小板,使人血小板表面P-选择素的表达增加(表达阳性率为90.4%)和PAC-1的结合量上升(阳性率为66.3%);终浓度为10、30 mg/L的TAP-SSL5可引起血小板的显著聚集。给小鼠单次尾静脉注射10 mg/kg的TAP-SSL5,可显著延长尾部出血时间,由对照组的(647.1±33.7)s延长至(753.6±127.3)s(P<0.01);而常规剂量组(3 mg/kg TAP-SSL5)尾部出血时间为(612.8±79.1)s,与对照组相比无显著差异(P>0.05)。结论融合蛋白TAP-SSL5保留了SSL5与血小板GPIbα结合的功能,有助于进一步提高TAP-SSL5的抗血栓作用,但同时也导致融合蛋白TAP-SSL5在高浓度情况下可延长出血时间和激活血小板。     

Effects of Aprotinin on Serum Interleukin-2 and Soluble Interleukin-2 Receptor during Cardiopulmonary Bypass

Cardiopulmonary bypass (CPB) impairs immunedefense function. It has been documented by a lot ofresearch that interleukin--2 and its receptor are of importance in regulating immunity responses. We observed the changes of interleukin--2 (IL--2) and soluble interleukin--2 receptor (IL--ZR) during heart valve(s) replacement operation and effects of aprotinin onthem.1 MATERIALS AND METHODSTwenty patients undergoing heart valve (s) replacement were randomly divided into two groups:control…     

抑肽酶对心脏手术血液保护量效关系的研究

目的 对抑肽酶的不同使用剂量和止血效果进行比较分析,探讨最佳应用方案。方法 ８２例心脏手术病人分为大剂量、半量和小剂量抑肽酶组,以不同的给药方案进行引流量、血制品用量、血红蛋白丢失量和血液抑肽酶浓度检测。结果 大剂量抑肽酶组与对照组相比,术后１２、２４ｈ引流量减少５４％、５５％,总红蛋白丢失量减少５６％。总血制品用量减少６５％。半量抑肽酶组可使术后１２、２４ｈ引流量减少４９％和５５％,与大剂量抑肽     

太湖珍宝口服液对黑腹果蝇寿命的影响

观察太湖珍宝口服液对黑腹果蝇寿命的影响。结果表明，０．５％、１．０％、２．０％太湖珍宝口服液均可明显延长果蝇的平均寿命与最高寿命，对果蝇的半数死亡期也有一定延长作用。     

Effect of aprotinin on the red cell immunity in cardiopulmonary bypass

Summary To investigate the effects of aprotinin on red cell (RC) immune function in the patients undergoing cardiopulmonary bypass (CPB), 20 patients who received valve replacement procedure were prospectively studied. The patients were randomly assigned to aprotinin group and control group. Red blood cell C₃b receptor ratio (RC₃bRR), red blood cell immune-component ratio (RICR), plasma C₃, C₄, CH₅₀ and IgG level were determined before operation, at the end of CPB and 1st, 3rd. 7th postoperative days. Our results showed that: (1) The blood requirement was reduced in aprotinin group. (2) After bypass, plasma C₃, C₄, CH₅₀ was maintained in aprotinin group, while they declined in control group. (3) Plasma level IgG fell in both groups, but at 7th postoperative day it recovered in aprotinin group while the level stayed at low level in control group. (4)The RC₃bRR and RICR was mildly inhibited in aprotinin group. It is concluded that the impairment of RC immune function caused by CPB can be mitigated by aprotinin, which may be related to the effects of aprotinin in blood sparing, restriction of complement activation, and reduction of blood requirement which could mitigate the non-specific inflammatory reaction. This project was supported by the grant form the National Fund of Natural Sciences (No. 39270658).     

Blood hibernation: a novel strategy to inhibit systemic inflammation and coagulation induced by cardiopulmonary bypass

Background Inflammation and coagulation are two intimately cross-linked defense mechanisms of most, if not all organisms to injuries. During cardiopulmonary bypass （CPB）, these two processes are activated and interact with each other through several common pathways, which may result in subsequent organ dysfunction. In the present study, we hypothesized that the addition of nitric oxide, prostaglandin E1 （PGE1）, and aprotinin to the systemic circulation, hereby referred to as blood hibernation, would attenuate the inflammation and coagulation induced by CPB. Methods Thirty adult mongrel dogs were equally divided into five groups, anesthetized and placed on hypothermic CPB （32℃）. Each group received respectively the following treatments： （1） inhalation of 40 ppm nitric oxide; （2） intravenous infusion of 20 ng.kgl.min1 of PGE1; （3） 80 000 kallikrein inhibitor units （KIU）/kg of aprotinin; （4） the combination of all three agents （blood hibernation group）; and （5） no treatment （control group） during CPB. Activation of leukocyte, platelet, endothelial cell, and formation of thrombin were assessed after CPB. Results As compared with the other four groups, leukocyte counts were higher, while plasma elastase, interleukin-8, CD11b mRNA expression, myeloperoxidase activities and lung tissue leukocyte counts were lower in the blood hibernation group （P 〈0.05 versus other four groups after CPB）. Plasma prothrombin fragment （PTF）1＋2, and platelet activation factors were lower, while platelet counts were higher in the blood hibernation group （P 〈0.05 versus other four groups at 6 and 12 hours after CPB）. Electron microscopy showed endothelial pseudopods protrusion, with cell adherence in all four groups except the blood hibernation group where endothelial cells remained intact. Conclusion Blood hibernation, effected by the addition of nitric oxide, PGE1 and aprotinin to the circulating blood during extra-corporeal circulation, was observed to attenuate the inflammation and coagulation induced by cardiopulmonary bypass, most likely by inhibiting the important common intermediates between the two cross-linked processes.