【作者】 欧阳锡林；

【导师】 刘景汉；

【作者基本信息】 中国人民解放军军医进修学院 ， 输血医学， 2010， 博士

【摘要】 迄今为止没有可供临床常规应用的血小板功能检测技术,国内外血液中心仅通过询问和捐血者问卷措施减少血小板功能缺陷的捐血者。血小板输注无效的临床发生率高达15%～40%,尚未见血小板制剂质量或（和）功能缺陷因素占血小板输注无效原因的统计报道。急需建立便捷、快速、价廉的血小板功能检测技术用于街头、献血屋和血站血小板捐血者的快速筛查。本研究探索以新型血小板激活剂——焦性没食子酸丙酯阳离子盐溶液（Cationic propyl gallate, c-PG）建立薄片法血小板凝集时间（Slide Platelet Clot Time, SPCT）,并初步应用于捐血人群、服用抗血小板药物患者、血小板减少症患者和各种血小板制品的快速筛查。第一部分焦性没食子酸丙酯阳离子溶液与SPCT的建立该部分主要通过选择的c-PG浓度、比较常用血小板激活剂诱导血小板凝集的可视性、检测血小板抑制剂对c-PG诱导的SPCT影响以及c-PG诱导的SPCT影响因素,优化建立SPCT。第一节c-PG浓度对其诱导的SPCT凝集时间的影响目的：筛选c-PG的合适浓度。方法：随机选取健康捐血者富含血小板血浆（PRP）8份,分别以终浓度为20μM、30μM、40μM和50μM的c-PG诱导SPCT。结果：（1）c-PG终浓度为20μM时SPCT为（174s±46s）,缓慢出现细小凝块或不凝；（2）c-PG终浓度为30μM时SPCT为（108s±21s）,出现凝集的时间约15s；（3）c-PG终浓度为40μM时SPCT为（68s±12s）,血小板凝块形成时间<4s；（4）c-PG终浓度为50μM或60μM时SPCT分别为（49s±8s）和（42s±6s）,血小板悬液从云雾状迅速转为清亮并出现凝块,血小板凝块很快形成。结论：随着c-PG浓度的增加,血小板开始发生凝集和形成明显的肉眼可见凝块的时间快速缩短,本研究选择c-PG终浓度为40μM应用于SPCT。第二节c-PG与常用血小板激活剂诱导的SPCT凝集强度和时间的比较目的：为SPCT选择可视性好的血小板激活剂。方法：随机选取20份健康捐血者PRP,分别测定添加终浓度为40μM c-PG、100μmol/L ADP、25μmol/LADP、3mol/L肾上腺素、1.5mol/L’肾上腺素、50mg／L胶原、25mg／L胶原、12mg／L胶原、1.2mg／ml瑞斯托霉素、500μg／ml花生四烯酸（AA）时,PRP的凝集强度和SPCT。结果：采用40μM c-PG、100μmol/L ADP, 50μmol/L ADP、25μmol/L ADP、3mol/L肾上腺素、1.5mol／L肾上腺素、50mg/L胶原、25mg/L胶原、12mg／L胶原、1.2mg/ml瑞斯托霉素、500μg／mlAA诱导PRP的凝集强度中位数和SPCT分别为为4+（45s±8s）、4+（45s±8s）、2+ （56s±13s）、2+（59s±19s）、2+（47s±10s）、2+（175 s±27s）±1+（108s±26s）、2+ （113s±24s）、1+（208s±52s）.0（＞300s）、2+（120s±24s）和3+（89s±19s）。结论：c-PG诱导PRP的凝块最大,SPCT时间较短,结果更易于判定。第三节血小板抑制剂对c-PG与常用血小板激活剂诱导的SPCT影响的比较目的：比较c-PG与常用血小板激活剂诱导的SPCT检测血小板抑制剂的能力。方法：随机选取15份健康献血者PRP,分别添加终浓度为10μM cAMP、20μg/ml PGI₂、0.001%EDTA和5%DMSO。分别测定终浓度为40μM c-PG, 100μmol/L ADP,3mol/L肾上腺素,100mg／L胶原,1.2 mg／L瑞斯托霉素,500μg/ml AA时的SPCT。结果：（1）40μM c-PG诱导添加10μM cAMP、20μg/ml PGI₂、0.01%EDTA和5%DMSO的SPCT分别为46s±7s、175s±21s、＞300s、>300s、116s±21s.>300s; （2） 100μmol/L ADP10μM cAMP、20μg/ml PGI₂、0.001%EDTA和5%DMSO的SPCT分别为,56s±11s、61s±19s、65s±17s、＞300s、>300s、>300s; （3）3mol/L肾上腺素10μM cAMP、20μg/ml PGI₂、0.001 %EDTA和5%DMSO的SPCT分别为,173s±27s、144s±36s*、130s±29s、＞300s、>300s、>300s; （4）50mg/L胶原诱导添加10μM cAMP、20μg/ml PGI₂、0.001%EDTA和5%DMSO的SPCT分别为,113s±24s、117s±22s、189s±31s、>300s、>300s、>300s; （5） 1.2 mg/L瑞斯托霉素诱导添加10μM cAMP、20μg/ml PGI₂、0.001%EDTA和5%DMSO的SPCT分别为120s±25s、170s±23s、294s±36s、>300s、218s±56、>300s; （6） 500mg/L AA诱导添加10μM cAMP、20μg/ml PGI2、0.001%EDTA和5%DMSO的SPCT分别为,90s±18s、137s±19s、180s±24s、＞300s、56s±47s、＞300s;结论：c-PG、瑞斯托霉素和AA均能检测出PGI2、cAMP、EDTA和DMSO对血小板的抑制作用。第四节c-PG、ADP和花生四烯酸诱导的SPCT检测志愿者服用阿司匹林前后血小板功能变化目的：比较c-PG、ADP和AA诱导的SPCT检测健康志愿者服用阿司匹林前后血小板功能变化的能力。方法：9名健康志愿者每天一次服用阿司匹林100mg,连续服用3天。采用终浓度为40μMc-PG溶液、20μmol/LADP、500μg/mlAA测定志愿者服用前后PRP的SPCT。结果：c-PG溶液、20μmol/LADP、500μg/mlAAc-PG和AA测定志愿者服用前后PRP的SPCT分别为42 s±12s和227s±26s（p=0.000）、56s±26s和89s±49s（p=0.038）、90s±23s和308s±13s（p=0.000）。诱导的血小板SPCT显著延长,而且AA已经诱导不出明显的凝集反应（p=0.000）；而ADP诱导的血小板SPCT时间在服用阿司匹林前后比较明显延长,但无显著差别（p=0.038）。结论：以40μM c-PG或500μg／mlAA为诱导剂的SPCT,均可高效检测出服用阿司匹林后患者血小板功能的显著变化。第五节肝素和枸橼酸盐对c-PG诱导的SPCT的影响目的：检测肝素和枸橼酸盐对c-PG诱导的SPCT的影响。方法：随机取PRP10份,分别加入终浓度为0,0.5,1,2,3U/L的肝素,109mmol/L,164mmol/L, 218mmol/L,272mmol/L的枸橼酸盐,测定SPCT。结果：分别添加OU／L、0.5U/L、1U/L、2U/L、3U/L的肝素或109mmol/L、164mmol/L、218mmol/L、272mmol/L的枸橼酸盐后PRP的SPCT分别为49s±8s、50s±9s、49s±8s、50s±10s.51s±9s、52s±8s、50s±7s、51s±9s和53s±11s。结论：添加肝素和明显增加枸橼酸盐浓度对c-PG诱导的SPCT无明显影响。第六节c-PG溶液4℃冰箱储存3年的稳定性研究目的：检测c-PG溶液储存3年的稳定性。方法：随机选取12份PRP,分别采用购买1年内和超过3年的c-PG溶液测定SPCT。结果：保存一年以内和保存超过三年的c-PG溶液诱导的SPCT分别为55s±1s6和55s±14s。结论：4℃储存三年对c-PG溶液诱导的血小板凝集时间无明显影响,c-PG具有良好的长期稳定性。第七节血小板计数和温度对cPG诱导的SPCT影响目的：研究血小板计数和温度对cPG诱导的SPCT影响。方法：1)随即取10份浓度调节好的健康献血者PRP倍比稀释为200×109／L,100×109／L,50x10⁹L,25×10⁹/L和13×10⁹/L,分别取样本测定SPCT; 2)随机10份PRP,分别置于40℃,30℃,20℃,10℃和0℃恒温金属浴恒温30 min,取样本进行SPCT测定；3)随机10份PRP室温（27℃）放置1h,2h,3h和4h,以及血小板振荡箱保存24h和48h,分别取样本测定SPCT。结果：10份PRP浓度调节为200x10⁹/L,100×10⁹/L,50×10⁹/L和25×10⁹/L,SPCT分别为32s±7s、67s±12s、99s±20s、1161s±41s和166s±27s,同一供血者血小板计数与其SPCT呈显著的直线负相关（r=0.910,p=0.005）；（2）40℃,30℃,20℃,10℃、5℃和0℃恒温金属浴恒温30 min的SPCT分别为44s±11s、42s±12s、43s±14s、60±12s、104s±20s和121s±27s,10℃、5℃和0℃保存30分钟与40℃,30℃,20℃保存有显著差异（p<0.01）；（3）室温放置1h,2h,3h,4 h,血小板振荡箱24h和48h SPCT分别为52s±8s、50s±7s、51s±9s、53s±11s（p=0.864）、59s±13s、87s±14s；室温放置4小时以内SPCT结果基本稳定,超过24小时SPCT明显延长。结论：（1）血小板计数与SPCT有明显的线性负相关。（2）20℃-40℃对SPCT无明显影响；而温度≤10℃时SPCT明显延长。（3）室温放置4小时对SPCT无明显影响。第二部分c-PG诱导的SPCT在捐血人群、患者和血小板制剂中的应用本研究测定全血捐献者和单采血小板捐献者、服用抗血小板药物的心内科患者、化疗致血小板减少症患者、单采血小板、浓缩血小板和深低温保存血小板的SPCT,探索其SPCT的主要特征。并模拟血小板输注的体外模拟混合SPCT,探索血小板输注无效的输血前筛查新方法。第一节.全血捐献者和单采血小板捐献者的SPCT目的：测定全血捐献者和单采血小板捐献者SPCT。方法：随机选取267名全血捐献者和相对固定的单采血小板的捐血者84人次,分离PRP测定SPCT。结果：267份全血捐献者和84份血小板捐献者SPCT分别为73s±21s（最大值216s,最小值为41s）和72±22s（最大值234s,最小值为38s）,两组无显著差异（p>0.05）；8例SPCT大于180秒,占2.5%。结论：351份全血或血小板捐献者SPCT平均值为73s,最大值234s,最小值为38s,血小板功能不良率为2.5%。第二节服用抗血小板药物的心内科患者目的：测定服用抗血小板药物治疗的心内科患者SPCT。方法：选取患有不稳定性心绞痛和冠心病的服用抗血小板药物的心内科住院患者65例,测定SPCT。结果：65例患者化疗后患者的SPCT均大于300秒。结论：40μM的c-PG不能诱导服用抗血小板药物连续超过3天患者的血小板在玻片上出现肉眼可见的凝集。第三节化疗后血小板严重减少患者目的：测定化疗后血小板严重减少患者SPCT。方法：选取血液科预约输注血小板治疗的化疗后血小板严重减少症患者34例,测定SPCT。结果：出现凝集标本5例（其中小于180秒者3例）,其余29例（85.3%）在超过300秒的计时内均未凝集。结论：当临床医生为化疗患者申请血小板输注治疗时,患者外周血血小板计数往往已经极度降低（多数患者<20×109／L）,这时患者多数不适合采用SPCT检测其血小板功能。第四节22℃液态振荡保存单采血小板和浓缩血小板的SPCT目的：测定单采血小板和浓缩血小板的SPCT。方法：1)随机选取单采血小板90份,测定浓度为200×10⁹/L、100×10⁹/L、50×10⁹/L、25×109/L、12.5×109／L时的SPCT；2)随机选取10份浓缩血小板,测定浓度为100×109／L、50×10⁹/L、25×10⁹/L、12.5×10⁹/L时的SPCT。结果：1)单采血小板浓度为200×10⁹/L、100×10⁹/L、50×10⁹/L、25×10⁹/L、12.5×10⁹/L时的SPCT分别为32s±7s、53s±14s、96s±29s、185s±50s;100例单采血小板中1例在计数为50×109／L时即不凝集,占1.0%；血小板计数为25×109／L时,SPCT为184s±49s,（中位数180s,最小值102s,最大值298s）,其中大于180s者49例（49.5%）；血小板计数为12.5×109／L时,凝集时间大于180s者16例（16.2%）,不凝集81例（81.8%）。2)10袋手工分浓缩血小板当计数为50×109／L时,SPCT为95s±31s（中位数99.5s,最小值50s,最大值142s）；计数为25×109／L时,SPCT为179s±58s（中位数169.5s,最小值118s,最大值281s）,其中大于180s者4例（40%）;血小板计数为12.5×109／L时凝集时间大于180s 10例（100%）,其中有5例不凝集（50%）。结论：血小板计数为50×109／L时,不出现凝集的单采血小板占1%,PC均出现凝集；血小板计数为25×109／L时,单采血小板SPCT大于180秒的占49.5%,PC为40%；血小板计数为12.5×109／L时,单采血小板SPCT大于180秒的占98%,PC为100%。第五节冰冻富含血小板血浆目的：测定冰冻PRP的SPCT。方法：随机选取40份健康捐血者的PRP,测定FPRP、DMSO-PRP和Cryo-PRP的SPCT。结果：FPRP、DMSO-PRP和Cryo-PRP的SPCT分别为59s±14s、108s±17s、115s±19s。与FPRP比较,DMSO-PRP和Cryo-PRP SPCT均发生了显著变化（p<0.001）; DMSO-PRP和Cryo-P（?）P比较SPCT也发生明显变化（p<0.005）。结论：由于含有5%DMSO并经过冰冻降温和复温过程,冰冻PRP的SPCT较FPRP比较显著的延长。第六节血小板输注有效性体外模拟混合SPCT实验目的：体外模拟血小板输注的混合SPCT实验。方法：随机选取申请血小板输注的血小板极度减少的血液科患者18例,分离患者PRP按照患者总血容量与所输机采血小板体积比混合,测定混合后的SPCT。记录输注血小板后的效果。结果：其中两例混合性SPCT大于180秒,且输注后两例患者出血倾向无明显改善或出血倾向继续加重,所以第二天继续申请输注血小板。另16利混合SPCT为79s±22s（最小值为46s,最大值为131s,中位数81.5s）,均无明显出血倾向加重或出血事故发生。结论：体外模拟混合SPCT检测可能用于血小板输注无效输血前筛查,但需要更进一步的临床资料证实。更多还原

【Abstract】 So far no platelet function assay is routing used in clinic so that the platelet donors with potential function defect are screened by consultative and questionnaire in blood center. Clinical platelet transfusion refractoriness is recurred in a high rate from fifteen percent to forty percent and the refractoriness due to platelet quality is not disclosed. Convenient, quickly, low cost platelet function assay is urgently required for platelet donor screening in the street, house donation and blood center. This study explores a new platelet agonist, cationic propyl gallate （c-PG） solution, for slide platelet clotting time （SPCT） and applying for platelet donors, patients with anti-platelet therapy, patients with thrombocytopenic, and platelet preparations.PART ONE, Cationic Propyl Gallate Solution and the Establishment of SPCTIn this part, the SPCT was optimized by choosing the concentration of c-PG, comparing the visibility of SPCT reduced by platelet agonists, monitoring the effectiveness of platelet inhibitors on SPCT reduced by platelet agonists, and the factors of SPCT reduced by c-PG.SECTION ONE, Effect of c-PG concentration on SPCTAIM Selecting a suitable concentrations of c-PG for SPCT. METHODS SPCT of eight random platelet-rich plasma （PRPs） reduced by 20μM、30μM、40μM和50μM c-PG was recorded. RESULTS （1） SPCT reduced by 20μM c-PG was 174s±46s and no clot or clot of platelets formatted slowly; （2） SPCT reduced by 30μM c-PG was 108s±21s and the clot formatted within 15s; （3） SPCT reduced by 40μM c-PG was 68s±12s and clot formatted within 4s; （4） SPCT reduced by 50μM or 60μM c-PG was 49s±8s and 42s±6s respectively and clot formatted very quickly. CONCLUSIONS With the increasing concentration of c-PG, time of SPCT reduced rapidly and platelet clot formatted quickly and 40μM c-PG is selected for SPCT. SECTION TWO, Comparison of the time and clot density of SPCT reduced by c-PG and other platelet agonistsAIM Selection of platelet agonists for good visibility of platelet clot in SPCT. METHODS The clot density and time of SPCT of twenty PRPs from health donors reduced by 40μM c-PG, 100μmol/L ADP,50μmol/L ADP,25μmol/L ADP, 3mol/L epinephrine,1.5mol/L epinephrine,50mg/Lcollagen,25mg/Lcollagen, 12mg/Lcollagen,1.2mg/ml ristocetin, and 500μg/ml arachidonic acid （AA） respectively were measured. RESULTS The median density and time of SPCT of twenty PRPs from health donors reduced by 40μM c-PG, 100μmol/L ADP, 50μmol/L ADP,25μmol/L ADP,3mol/L epinephrine,1.5mol/L epinephrine, 50mg/Lcollagen,25mg/Lcollagen,12mg/Lcollagen,1.2mg/ml ristocetin, and 500μg/ml AA respectively were 4+（45s±8s）,4+（45s±8s）,2+（56s±13s）, 2+（59s±19s）,2+（47±10）s,2+（175±27）, 1+（108s±26s）,2+（113s±24s）, 1+（2083±52s）,0（> 300s）,2+（120s±24s）, and 3+（89s±19s）, respectively.CONCLUSION SPCT reduced by 40μM c-PG has a short aggregation time and best clot density.SECTION THREE, Comparison of the effect of platelet inhibitors on the SPCT induced by c-PG and platelet agonistsAIM Comparison of the efficacy of platelet inhibitors on SPCT reduced by c-PG and platelet agonists. METHOD Time of SPCT of fifteen random PRP from healthy donors with a adjunction of 10μM cAMP,20μg/ml PGI₂,0.01%EDTA, 5%DMSO, and 0.025% NaAzide, respectively reduced by 40μM c-PG, 100μmol/L ADP,3mol/L epinephrine,50mg/Lcollagen,1.2 mg/L ristocetin, 500μg/ml AA respectively were measured. RESULT （1） Time of SPCT of fifteen PRPs control and that with a adjunction of cAMP, PGI₂, EDTA, DMSO, and NaAzide, respectively reduced by 40μM c-PG were 46s±7s,175s±21s,>300s, >300s,116s±21s, and>300s respectively;（2） Time of SPCT of fifteen PRPs control and that with a adjunction of cAMP, PGI₂, EDTA, DMSO, and NaAzide respectively reduced by 100μmol/L ADP were 56s±11s,61s±19s,65s±17s,> 300s,>300s, and>300s respetively;（3） Time of SPCT of fifteen PRPs control and that with a adjunction of cAMP, PGI₂, EDTA, DMSO, and NaAzide respectively reduced by 3mol/L Epinephrine were 173s±27s,144s±36s, 130s±29s,>300s,>300s, and>300s respectively; （4） Time of SPCT of fifteen PRPs control and that with a adjunction of lOμM cAMP,20μg/ml PGI₂, 0.01%EDTA,5% DMSO, and NaAzide respectively reduced by 50mg/LCollagen were 113s±24s,117s±22s,189s±31s,>300s,>300s, and>300s respectively; （5） Time of SPCT of fifteen PRPs control and that with a adjunction of cAMP, PGI₂, EDTA, DMSO, and NaAzide respectively reduced by 1.2 mg/L ristocetin were 120s±25s,170s±23s,294s±36s,>300s,218s±56, and>300s respectively; （6） Time of SPCT of fifteen PRPs control and that with a adjunction of cAMP, PGI₂, EDTA, DMSO, and NaAzide, respectively reduced by 500mg/l AA were 90s±18s,137s±19s,180s±24s,>300s,156s±47s, and>300s respectively;CONCLUSION SPCT reduced by c-PG, ristocetin or AA is affected by PGI₂, cAMP, EDTA, DMSO or NaAzide.SECTION FOUR, SPCT reduced by c-PG, ADP or AA monitoring the platelet function of volunteers ingestion of acetyl sal icylic acid （ASA）AIM Comparison of SPCT reduced by c-PG, ADP or AA monitoring the platelet function of volunteer ingestion of ASA. METHOD Nine healthy volunteers took 100mg ASA once a day for three days straight. SPCT of volunteers before and after ingestion of ASA reduced by 40μM c-PG,20μmol/LADP, or 500μg/ml AA were measured. RESULT SPCT of volunteers reduced by c-PG, ADP, or AA before and after ingestion of ASA were 42s±12s and 227s±26s （p=0.000）,56s±26s and 89s±38s （p=0.038）,93s±23s and 308s±13s （p=0.000）, respectively. CONCLUTION SPCT reduced by 40μM c-PG or 500μg/ml AA could effectively monitor the platelet function damage after ingestion of ASA.SECTION FIVE, The affect of adjunction of heparin or citrate on SPCT reduced by c-PGAIM The affect of PRP adjunction of heparin or citrate on SPCT reduced by c-PG. METHODS SPCT of ten random PRPs with a adjunction of 0,0.5,1,2,3U/L heparin or 109mmol/L,164 mmol/L,218mmol/L,272mmol/L citrate were measured. RESULRs SPCT of ten random PRPs with a adjunction of heparin or citrate were 49s±8s,50s±9s,49s±8s,50s±10s,51s±9s,52s±8s,50s±7s,51s±9s, and 53s±11s. CONCLUTION Adjunction of heparin or citrate has no significant affect on SPCT reduced by c-PG.SECTION SIX, Study on the stability of more than three years four dgree C storageAIM Study on the stability of c-PG. METHODS SPCT of twelve random PRPs reduced by c-PG with a storage within one year and c-PG with a storage more zhan three years respectively were measured. RESUNLTs SPCT of twelve random PRPs reduced by c-PG with a storage within one year and c-PG with a storage more zhan three years respectively were 55s±16s and 55s±14s （p>0.05）. CONCLUSION c-PG has a character of long term stability.SECTION SEVEN, Affect of platelet count and temperature on SPCT reduced by cPGAIM Stuy on the affect of platelet count and temperature on SPCT reduced by cPG. METHODs （1） Ten random PRPs were diluted into count of 200×10⁹/L, 100×109/L,50×109/L,25×109/L和13×109/L and then the SPCT of that were measured; （2） SPCT of ten random PRPs treated at 40degree C,30 degree C,20 degree C,10 degree C or 0 degree C in meltal bath for 30 min were measured; （3） SPCT of ten random PRPs treated at room temperature for lh,2h,3h,4h,24h, and 48h were measured. RESULTs （1） SPCT of ten random PRPs wich were diluted to 200×109/L,100x109/L,50×109/L,25×10⁹/L,13×10⁹/L, and 6×109/L were 54s±11s,80s±14s,110s±17s,137s±23s, and 166s±27, respectively; Platelet count and its SPCT of the same individual showed a significant negative correlation （r=0.946, p=0.004）; （2） SPCT of ten random PRPs treated at 40degree C,30 degree C,20 degree C,10 degree C or 0 degree C in meltal bath for 30 min were 44s±11s,42s±12s,43s±14s,60±12s,104s±20s, and 121s±27s （p<0.01）; （3） SPCT of ten random PRPs treated at room temperature for 1h,2h,3h,4h,24h, and 48h were 52s±8s,50s±7s,51s±9s,53s±11s,59s±11s, and 87s±14s, respectively. CONCLUSION （1） Platelet count and its SPCT of the same individual showed a significant negative correlation （r=0.910, p=0.005）; （2） Time of SPCT was prolonged when PRPs were treated below ten degree C for thirty minutes; （3） Time of SPCT was not affected by the treatment of four hours room temperature storage.PART TWO, Application of SPCT reduced by c-PG for platelet donors, patients and plat let preparationThis study on the measurement of SPCT of whole blood donors, apheresis platelet concentration donors, cardiology patients with anti-platelet therapy, thrombocytopenia induced by chemotherapy, apheresis platelet, platelet concentrations, and cryopreserved platelet. Effectiveness of platelet transfusion was simulated by SPCT of in vitro mixture of donor’s PRP and patient’s PRP.SECTION ONE, SPCT of whole blood donors and apheresis platelet donorsAIM Measurement of SPCT of whole blood donors and apheresis platelet donors. METHODs SPCT of PRPs from 267 whole blood donors and 84 apheresis platelet donors were measured. RESULTs SPCT of PRPs from 267 whole blood donors and 84 apheresis platelet donors were 73s±21s（41s-216s） and 72s±22s （38s～234s） （p>0.05）; SPCT of five whole blood donors and three apheresis platelet donors exceed 180s. CONCLUSIONS The mean of SPCT of 351 blood donors is 73s（38s～234s）; Platelet dysfunction rate of blood donors is about 2.5 percent （8/351）.SECTION TWO, SPCT of cardiology patients with ant i-platelet therapyAIM Measurement of SPCT of cardiology patients with anti-platelet therapy. METHODS SPCT of sixty five random cardiology patients with anti-platelet therapy were measured. RESULTs SPCT of sixty five cardiology patients with anti-platelet therapy were all more than three hundred seconds. CONCLUSION The platelet clot can not be induced by 40μM c-PG in the cardiology patients with more than three days anti-platelet therapy. SECTION THREE, SPCT of patients with thrombocytopenia i nduced by chemotherapy.AIM Measurement of SPCT of patients with thrombocytopenia induced by chemotherapy. METHODs SPCT of thirty four random hematological patients with severe thrombocytopenia induced by chemotherapy who were made a reservation for platelet transfusion. RESULTs Platelet clot of five patients formatted and that of twenty nine （85.3 per cent） were not emerged when the time counting reach to three hundred seconds. CONCLUSION SPCT is not so suitable for monitoring platelet function of the most of patients with severe thrombocytopenia （less than 20×10⁹/L）.SECTION FOUR, SPCT of aphaeresis platelet and platelet concentration （PC）AIM Measurement of SPCT of aphaeresis platelet and PC. METHODS 1) SPCT of one hundred random aphaeresis platelet were diluted into 200×10⁹/L, 100×10⁹/L,50×10⁹/L,25×10⁹/L, and 12.5×10⁹/L, respectively were measured; 2) SPCT of ten random PC were diluted into 200×10⁹/L,100×10⁹/L,50×10⁹/L, 25×10⁹/L, and 12.5×10⁹/L, respectively were measured. RESULTs （1） SPCT of one hundred random aphaeresis platelet were diluted into 200×10⁹/L,100×10⁹/L, 50×10⁹/L,25×10⁹/L, and 12.5×10⁹/L, respectively were 32s±7s,53s±14s,96s±29s, and 185s±50s, respectively; One aphaeresis platelet cannot occur platelet clot when the count is 50 X 109/L; SPCT is 184s±49s when the platelet count is 25 X 109/L and that of 49 （49.5%）aphaeresis platelet is more than 180s; SPCT of only one aphaeresis platelet is less than 180s when the count is 12.5 X 109/L. （2） SPCT of ten PC is 95s±31s when the count is 50×10⁹/L; SPCT is 179s±58s when the count is 25×109/L and that of four PC is more than 180s; SPCT of all PC is more than 180s when the count is 12.5 X 109/L. CONCLUSIONS When the count is 50×109/L, SPCT of one percent of aphaeresis platelet is more than 180s; That of 49.5 percent of aphaeresis platelet and 40 percent of PC is more than 180s when the count is 25×109/; That of 98 percent of aphaeresis platelet and 100 percent of PC is more than 180s when the count is 12.5 X 109/L.SECTION FIVE, Cryopreserved fresh platelet-rich plasmaAIM Measurement of SPCT of Cryopreserved fresh platelet-rich plasma. METHODS SPCT of forty random FPRP, FPRP with 5 percent DMSO （DMSO-PRP）, and frozen and thawed PRP （Cryo-PRP） were measured. RESULTS SPCT of FPRP, DMSO-PRP, and Cryo-PRP is 59s±14s,108s±17s, and 115s±19s, respectively. Compared with SPCT of FPRP, DMSO-PRP and Cryo-PRP was significantly prolonged （p< 0.001）. CONCLUSION SPCT of cryopreserved platelet is 115s±19s, significantly prolonged by DMSO and the process of freezing and thawing.SECTION SIX, Platelet transfusion was simulated by SPCT of ex vivo mixture of platelet preparation and patient’s PRPAIM Explore the simulation of SPCT of ex vivo in vitro mixture of donor’s PRP and patient’s PRP. METHODS PRP of eighteen random hematological patient’s and the platelet preparation were mixed in vitro simulated clinic platelet transfusion. The SPCT of the mixture were measured and the effectiveness of the platelet transfusion were recorded. RESULTS SPCT of in vitro mixture of two cases were more than 180s. The two patients suffered with not improved bleeding intendancy or increased bleeding intendancy after transfused platelet preparation and another platelet preparation was reserved for them. The SPCT of the other sixteen mixture were 79s±22s （46s～131s）. No significant increased bleeding intendancy or bleeding accident occurred in the sixteen cases. CONCLUSION SPCT of the mixture may be used for screening platelet transfusion refraction before platelet transfused but forther clinic trail must be done for more envidence.更多还原