现居东莞人群人类血小板1-6抗原系统基因分型研究

目的研究现居东莞人群人类血小板抗原（human platelet antigen, HAP） 1-6系统基因分型及其多态性分布特征。方法用DNA提取试剂盒提取外周血标本中的DNA；用聚合酶链式反应一序列特异性引物（PCR-SSP）扩增HPA等位基因。结果80名健康、无血缘关系的现居东莞人群每个样本均检测到HPA-1a、2a、4a、5a、6a基因；HPA-4a、5a呈现纯合子单态性，未检测出相应的等位基因HPA-b；对于HPA-1、-2、-6主要以a／a纯合子为多，a／a基因型频率分别是0．975、0．725、0．9625，没有b／b纯合子出现。结论HPA血型系统多态性具有人群特征。与其他种族相比，现居东莞人群HPA血型系统中a基因频率较高，b基因频率较低，提示在现居东莞人群中由b基因不合所引起的同种免疫病理反应要低于其他种族人群。现居东莞人群HPA-1、-2、-3、-6系统具有多态性，HPA-2、-3抗原杂和率比较高，提示在临床输血实践中可能具有重要的免疫学意义。同时，在此次研究数据的基础上建立了现居东莞人群血小板基因频率数据库和血小板基因型已知供者库。     

福建汉族人群人类血小板抗原HPA-15多态性分析(附113例报告)

目的分析福建汉族人群人类血小板抗原(HPA)-15基因多态性,为临床血小板输注提供依据。方法采用聚合酶链反应-序列特异引物法(PCR-SSP)技术对113名福建汉族健康个体进行HPA-15基因分型。结果HPA-15aa、HPA-15ab及HPA-15bb的分布频率分别为0.274、0.522和10.204,与我国其他省份汉族人群比较,差异无统计学意义。HPA-15a、HPA-15b抗原不配合率分别为0.116和10.133。结论HPA-15基因多态性区域差异不明显。HPA-15a/b抗原不配合率较HPA其他系统高,可能导致产生同种免疫性血小板抗体的概率增加,在临床血小板输注中的作用值得重视。     

患者HPA 1-6,9,15基因多态性对血小板输注无效的临床意义

目的观察人类血小板特异性抗原(human platelet alloantigens,HPA)1-6,9,15系统及其基因多态性与血小板输注无效(platelet transfusion refractoriness,PTR)的研究。方法选取40例血小板输注无效患者的外周血标本和供者标本,对其进行血小板特异性糖蛋白抗体检测,根据血小板回收率评判血小板输注效果,采用聚合酶链式反应(PCR)结合直接测序方法,对供、患者进行HPA1-6,9,15抗原系统基因进行分型,动态观察患者同型血小板输注后血小板恢复百分率(percent platelet recovery,PPR),探讨HPA基因多态性和血小板输注无效的关系。结果患者和供者HPA-1、4、9均未检测出HPA-b基因,呈呈aa纯合子单态性分布;HPA-5、6则主要以aa纯合子为多,较少看到bb纯合子。而HPA-2、3、15呈明显多态性分布,HPA-2、3、5、6、15等位基因频率均呈多态性分布。结论对反复多次发生血小板输注无效的患者,除需考虑HLA基因相关外,还与HPA基因多态性相关。在做HPA配型时,多数人只需检测供者与患者HPA2、3、15基因就可以显著减少血小板输注无效的发生。     

山西地区血液病患者人类血小板抗原基因分型研究

目的:系统分析山西地区血液病患者人类血小板抗原(HPA)基因分型及其多态性,为HPA基因库建设提供实验数据和理论依据,为本地区血小板输注无效(PTR)患者的血小板(PLT)有效输注建立理论和临床基础。方法:选取血液病住院患者97例,分为输注有效组(52例)和输注无效组(45例),提取全血DNA,采用聚合酶链反应-序列特异性引物技术(PCR-SSP)进行HPA-1~6,15抗原系统基因分型。结果:输注有效组HPA-1,4,5,6主要以aa型为主,HPA-3和HPA-15杂合度较高。输注无效组HPA-1,4,5,6主要以aa型为主,HPA-2,HPA-3,HPA-15杂合度较高。结论:山西地区人类血小板同种抗原血型系统基因多态性表现出人群特点。人类血小板抗原血型中a基因频率明显较高;血液病患者HPA-2,HPA-3,HPA-15抗原对配偶不配合比例高。提示临床治疗中对于反复输注PLT的血液病患者需高度关注HPA-2,HPA-3,HPA-15基因型,并对其进行检测,尽量同型输注,避免产生血小板同种抗体,引起血小板输注无效。     

广西壮族自治区壮族居民血小板抗原HPA-2基因分布频率研究

目的：探讨广西壮族自治区壮族居民HAP-2基因分布频率,指导相关血小板输注的治疗决策。方法采用聚合酶链式单核苷酸多态性方法对2659名广西壮族自治区壮族健康成年人的血液样本进行HPA-2基因分型检测,用直接测序法解决HPA-2分型差异。结果 HPA-2a/2a、HPA-2a/2b和HPA-2b/2b的等位基因频率分别为0.9157、0.0824和0.0019;测得基因多态性位点与数据库收录信息全部吻合,未发现新的单核苷酸多肽性（SNP）位点。广西壮族自治区壮族居民HPA-2a/2b基因的杂合程度与大多数亚洲国家的研究结果相似,与非洲人相比存在显著差异。结论该研究有助于确定该地区居民HPA-2多态性分布及其潜在临床意义,能更好地理解和治疗壮族免疫性介导的血小板疾病。     

贵阳地区汉族人群人类血小板抗原1～5,15基因多态性与2型糖尿病合并周围神经病变的相关性研究

目的探讨人类血小板抗原1～5,15(HPA-1～5,15)基因多态性与贵阳市汉族2型糖尿病合并周围神经病变的发生是否具有相关性。方法 2型糖尿病合并周围神经病变患者75例(DPN组),不合并周围神经病变患者24例(非DPN组),健康体检者100例(对照组)。采用酚/氯仿方法提取DNA,用多聚酶链反应-序列特异性引物方法进行基因组分型。结果 DPN组、非DPN组的HPA-1aa、2aa、3aa型基因频率均明显高于对照组,而DPN组与非DPN组之间差异无显著性;DPN组、非DPN组的HPA-1a、2a、3a等位基因频率明显高于对照组,而DPN组与非DPN组之间差异无显著性。HPA-4、5、15的基因型、等位基因频率在对照组、DPN组与非DPN组之间比较差异均无显著性。结论 HPA-1～5,15基因多态性与贵阳市汉族人群2型糖尿病合并周围神经病变的发生可能无关。     

绍兴市2012—2020年流失单采血小板献血者召回策略探讨

目的 调查单采血小板献血者的流失原因，探讨流失献血者召回策略。方法 绍兴市中心血站2012年1月1日至2020年12月31日成功捐献单采血小板的献血者3 446人，筛选出流失单采血小板献血者1 860人（54.0%）。比较流失与固定单采血小板献血者在人群特征上的差异，回访调查流失原因及血小板捐献再动员，分析流失后成功召回情况。结果 相较于固定单采献血者，流失单采血小板献血者中非本地籍、女性、18～40岁、初中及以下文化程度、学生占比较高。流失单采血小板献血1 860人中召回后再捐献145人，召回率7.8%。单采血小板献血者流失原因中因“主观意愿不愿意再献”“没时间或交通不便”“工作人员联系不上或已离开本地区”的占比分别为11.5%、26.4%、35.2%，召回成功率为2.3%、14.3%、0.5%，因“工作人员联系少或没联系”占2.2%，召回成功率最高（62.5%）。流失单采血小板献血者召回后再捐献人群中，本地籍、捐献次数> 5次者召回成功率明显较高。结论 流失单采血小板献血者重新召回需要充分了解发生流失的原因，针对不同需求采取相应的措施，实现定向再招募召回的目的。     

基于精准招募策略的单采血小板献血人群特征及采供情况分析

目的 了解基于精准招募策略下顺德区中心血站(以下简称“本站”)单采血小板献血人群特征及采供情况,为进一步完善单采血小板献血者精准招募的长效机制提供科学依据。方法 选取2020年1月～2022年12月的620名新增单采血小板献血者和5446人次单采血小板捐献的献血者资料,以及单采血小板的采集总量、供应总量作为观察组;2017年1月～2019年12月的461名新增单采血小板献血者和4663人次单采血小板捐献的献血者资料,以及单采血小板的采集总量、供应总量作为对照组。对照组采用传统宣传招募方式进行招募,观察组在传统宣传招募方式的基础上引入精准招募策略进行招募。比较两组单采血小板献血者献血人次的特征(包括文化程度、职业、年龄、性别),新增单采血小板献血者的特征(包括职业、年龄、性别),单采血小板采集、供应情况(包括单采血小板采集总量、血小板供应总量、血小板采供比)。结果 两组单采血小板献血者献血人次的文化程度、职业、年龄比较,差异具有统计学意义(P<0.05);两组单采血小板献血者献血人次的性别比较,差异无统计学意义(P>0.05)。两组新增单采血小板献血者的职业、年龄、性别比较,...     

广州市单采血小板输血相关传染病筛查不合格回顾性研究

目的探讨来自无偿和互助献血单采血小板血液安全性,为完善单采血小板招募策略提供理论依据。方法 2014年1月～2018年3月共采集单采血小板献血者样本157 692份,使用两种不同厂家ELISA试剂盒检测HBsAg、抗-HCV、抗-HIV、抗-TP及ALT(速率法),同时进行HBV DNA、HCV RNA、HIV RNA核酸检测(NAT),比较无偿和互助单采血小板筛查总体不合格率及各筛查项目的不合格情况,并对单试剂阳性献血者进行追踪回访,比较不同来源献血者解锁率。结果 2014年1月～2018年3月共检测单采血小板献血者样本157 692份,共检出1495份不合格样本,总体不合格率为0.95%,广州市无偿和互助单采血小板血液筛查不合格率均呈下降趋势,互助单采血小板总体和各年不合格率均高于无偿献血(P 0.05),互助献血者单采血小板血液筛查项目:HBsAg、抗-HCV、抗-HIV(单试剂阳性)、抗-TP、ALT、单项NAT的阳性率均高于无偿献血者。对404位单试剂阳性单采献血者进行追踪回访,无偿单采血小板献血者解锁率为82.37%;互助单采血小板献血者解锁率为70.65%(χ2=6.03,P 0.05)。结论单采血小板互助献血血液安全风险较无偿献血高,不适宜作为今后单采血小板的招募发展方向,采供血机构应坚持从固定无偿献血者中招募,以保障临床用血安全,做好单试剂阳性献血者复检和归队工作,扩大单采血小板献血者队伍。     

金华市2020—2022年单采血小板无偿献血者人群特征及血液检验不合格情况分析

目的 分析金华市2020—2022年单采血小板无偿献血者人群特征及血液检验不合格情况。方法 选取金华市中心血站2020年1月至2022年12月参加单采血小板无偿献血者临床资料,分析不同性别、年龄、职业及学历的人群特征条件下血液检验不合格情况,血液检验项目包括谷丙转氨酶（GPT）、梅毒螺旋体（TP）、人类免疫缺陷病毒（HIV）、乙型肝炎病毒表面抗原（HBsAg）及病毒核酸检测。结果 2020—2022年单采血小板无偿献血者血液检验不合格率为0.3%(23/8 671),血液检验不合格率呈现先增加后降低趋势。不同血液检验项目中,TP、HBsAg不合格率高于GPT、HIV及核酸不合格率。男性血液检验不合格率（0.34%,22/6 448）高于女性（0.04%,1/2 223）,18～35岁血液检验不合格率高于其他年龄段献血者,且TP检验不合格率最高。农民献血者血液检验不合格率高于其他职业的献血者,且GPT、TP及HBsAg检验不合格率最高。初中及以下单采血小板无偿献血者血液检验不合格率高于其他学历献血者,且HBsAg检验不合格率最高。结论 2020—2022年金华市参加单采血小板无偿献血人群...     

Frequency distribution of phenol sulfotransferase 1A1 activity in platelet cells from healthy Japanese subjects.

AIMS: To determine the distribution of sulfotransferase 1A1 (SULT1A1) activities, we used trans-4-hydroxytamoxifen (OHT) as a substrate to test samples from a Japanese population to examine whether the SULT1A1*2 allele can account for the wide distribution of OHT sulfating activity. We also studied genetic mutations other than the SULT1A1*2 allele to determine the cause of differences in SULT1A1 protein expression and activity. METHODS: The subjects were 103 healthy Japanese adults. Identification of SULT1A1 genotypes was performed using a polymerase chain reaction-restriction fragment length polymorphism method. SULT1A1 activity in platelet cytosol was assayed using OHT as a substrate. SULT1A1 protein was detected using Western blotting analysis. Mutations other than SULT1A1*2 in the SULT1A1 gene were detected using sequencing analysis. RESULTS: SULT1A1*2 allele frequency was found to be 16.5%, while SULT1A1 activity ranged from 63 to 1860pmol sulfated/h/mg platelet protein (260+/-241pmol sulfated/h/mg platelet protein, median+/-S.D.) using OHT as a substrate. The median values in subjects with SULT*1/*2 (221+/-113pmol sulfated/h/mg platelet protein, range 63-442, n=26) and SULT*2/*2 (124+/-66pmol sulfated/h/mg platelet protein, range 74-231, n=4) were significantly lower than that in subjects with SULT*1/*1 (303+/-267pmol sulfated/h/mg platelet protein, range 97-1859, n=73). A novel G148C mutation was found in one subject, who showed the lowest OHT sulfating activity, for a frequency of 0.49%. CONCLUSION: There was wide variety of OHT sulfating activities found among the present healthy Japanese subjects. The SULT1A1*2 allele was found to be a common variant allele and was associated with decreased OHT sulfating activity. These observations may be related to inter-individual variations of OHT pharmacokinetics and the pharmacologic effects of tamoxifen seen in Japanese patients with breast cancer.     

血小板抗体产生对血小板输注效果的影响

目的观察血小板抗体产生对血小板输注效果的影响。方法选择本院100例输血患者分为两组,A组患者输注的血小板没有滤除白细胞,B组患者输注的血小板滤除白细胞。检测两组血小板相关抗体（HLA抗体）阳性率和血小板特异性抗体（HPA抗体）阳性率,评估两组血小板输注效果。结果 A组血小板输注无效率为48.9%,B组血小板输注无效率为12.7%,差异有统计学意义（P〈0.01）。两组HLA抗体阳性率分别为55.6%和14.5%,差异有统计学意义（P〈0.01）。两组HPA抗体阳性率是15.6%和10.9%,差异无统计学意义（P〉0.05）。结论输注滤除白细胞的血小板,可以提高血小板的输注效果。     

Hepatic platelet accumulation in Fas-mediated hepatitis in mice

Platelets are reported to be causally involved in experimental hepatitis. Jo2, an agonistic anti-Fas antibody, induces hepatitis in mice. We examined the in vivo behaviors of platelets in mice injected with this antibody (analyzed by measuring 5-hydroxytryptamine, a constituent of platelets). We found that Jo2 induces platelet accumulation predominantly in the liver, and that this hepatic platelet accumulation (HPA) precedes the increases in hepatitis markers (alanine- and asparagine-aminotransferases [ALT and AST]). By electron microscopy, we detected entry of platelets into hepatocytes, and also evidence of apoptosis among hepatocytes. A caspases-3/6/7/8/10 inhibitor prevented the Jo2-induced HPA and hepatitis. In platelet-depleted mice, contrary to our expectations, the Jo2-induced hepatitis was not reduced, and actually the increase in AST was significantly augmented, although the survival time of mice given a lethal dose of Jo2 was significantly increased (nearly doubled). Interestingly, prior induction of HPA by a low dose of lipopolysaccharide markedly reduced Jo2-induced hepatitis. Jo2 also induced HPA and hepatitis in mice deficient in both IL-1 and TNFα, although Jo2 increased the blood level of TNFα in wild-type mice. These results suggest that in Jo2-induced hepatitis: (i) platelets accumulate predominantly in the liver as a result of hepatic lesions, and that this precedes the release of transaminases from hepatocytes, and (ii) IL-1 and TNFα are not essential for Jo2-hepatitis. We hypothesize that platelet accumulation in the liver may, contrary to our expectations, be protective when the hepatitis is local or not severe, but harmful when hepatitis is severe.     

Effect of single-dose sertraline on the hypothalamus-pituitary-adrenal system, autonomic nervous system, and platelet function

OBJECTIVE: Pharmacological treatment with selective serotonin reuptake inhibitors (SSRIs) is thought to decrease coronary risk in patients with depressive disorder. Selective serotonin reuptake inhibitor intake may (1) attenuate the hypothalamus-pituitary-adrenal (HPA) system, (2) improve disturbances of the autonomous nervous system, and (3) dampen the aggregability of platelets. There is only limited information about the influence of acute treatment with SSRIs on these systems, which is especially important for the initiation of therapy in high-risk cardiac patients. We compared the reaction of these systems to physical stress with single-dose SSRI treatment (100 mg) with that of placebo treatment. METHODS: Using a double-blind, crossover, placebo-controlled design, we assessed HPA system activity via serum cortisol and corticotropin as well as sympathetic nervous system by determining serum norepinephrine and epinephrine levels at baseline and as a response to stress. Analysis of heart rate variability (HRV) provided information on sympathetic/parasympathetic balance. Platelet activity was measured via flow-cytometric determination of platelet surface activation markers along with the serotonin (5-HT) uptake of platelets. RESULTS: We studied 12 healthy young men under placebo and verum conditions. We found higher HPA system activity at baseline and after physical activity under sertraline when compared with placebo, no difference in sympathetic nervous system activity after physical exertion and only slightly heightened baseline epinephrine values after sertraline intake. No difference was seen between sertraline and placebo intake regarding platelet activity and 5-HT uptake, HRV, blood pressure, and HR. CONCLUSIONS: Initiating sertraline treatment increases HPA system activity and epinephrine concentrations. We found no clinically relevant effect of single-dose sertraline treatment on autonomous nervous function, platelet activity, or platelet 5-HT uptake. These findings may not be extrapolated to patients with affective or cardiac disorders or to other SSRIs.     

Cilostazol and dipyridamole synergistically inhibit human platelet aggregation

It has been previously shown that cilostazol (Pletal), a drug for relief of symptoms of intermittent claudication, potently inhibits cyclic nucleotide phosphodiesterase type 3 (PDE3) and moderately inhibits adenosine uptake. It elevates extracellular adenosine concentration, by inhibiting adenosine uptake, and combines with PDE3 inhibition to augment inhibition of platelet aggregation and vasodilation while attenuating positive chronotropic and inotropic effects on the heart. In the present study, we tested the hypothesis that cilostazol combined with a more potent adenosine uptake inhibitor, dipyridamole, synergistically inhibited platelet aggregation in human blood. In the presence of exogenous adenosine (1 microM), the combination of cilostazol and dipyridamole synergistically increased intra-platelet cAMP. Furthermore, cilostazol inhibited platelet aggregation in a washed platelet assay concentration-dependently with IC50s of 0.17 +/- 0.04 microM (P < 0.05 versus plus adenosine alone of 0.38 +/- 0.05 microM), 0.11 +/- 0.06 microM (P < 0.05), and 0.01 +/- 0.01 microM (P < 0.005) when combined with 1, 3, or 10 microM dipyridamole, respectively (n = 5). In whole blood, cilostazol (0.3 to 3 microM) and dipyridamole (1 or 3 microM) synergistically inhibited collagen- and ADP-induced platelet aggregation in vitro. Furthermore, the synergism was confirmed in an open-label, sequential study in healthy human subjects using ex vivo whole-blood collagen-induced platelet aggregation. Four hours after oral co-administration of cilostazol (100 mg) and dipyridamole (200 mg), platelet aggregation was inhibited by 45 +/- 17%, while no significant inhibition was observed from subjects treated with either drug alone. The combination may provide a potential treatment of arterial thrombotic disorders.     

PON1(rs662)基因多态性对冠脉介入治疗患者血小板高反应性的影响

目的：探讨PON1（rs662）基因多态性对冠脉介入治疗患者血小板高反应性的影响。方法：279例PCI术后患者,给予氯吡格雷75 mg·d^-1及阿司匹林100 mg·d^-1抗血小板治疗,服用5 d后空腹抽取静脉血,并检测血栓弹力图（TEG）。将氯吡格雷治疗后血小板高反应性（HPR）定义为二磷酸腺苷（ADP）诱导的血小板抑制率<50%。应用单因素和多因素回归分析PON1（rs662）基因多态性及临床因素对HPR的影响,利用受试者工作曲线（ROC curve）检验联合独立风险因素模型预测HPR的效力。结果：279例PCI术后患者HPR发生率为19.0%。多因素logistic回归分析示PON1（rs662） A等位基因（OR=2.101,95% CI=1.002~4.406,P=0.049）、糖尿病（OR=1.921,95% CI=1.023~3.604,P=0.042）为HPR的独立风险因素。联合独立风险因素模型预测HPR的曲线下面积（AUC）为0.680（95% CI=0.600~0.760,P<0.001）。结论：PON1（rs662） A等位基因及糖尿病为PCI术后患者HPR的独立危险因素。通过联合独立风险因素模型预测HPR的效力良好。     

Resveratrol attenuates thromboxane A2 receptor agonist-induced platelet activation by reducing phospholipase C activity

Resveratrol (3,5,4'-trihydroxystilbene) is a naturally occurring compound shown to decrease the incidence of thromboembolic disease. Although considerable data are available as to the inhibitory effect of resveratrol on the platelet aggregation and thrombopoiesis in human, its underlying mechanism, at the cellular level, has not been rigorously studied. In this experiment, we studied the effect of resveratrol and 1-[6-[[17-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione, a phospholipase C inhibitor (U-73122) on the thromboxane A2 receptor agonist (9,11-dideoxy-11 alpha,9 alpha-epoxymethanoprostaglandin F(2 alpha), U46619)-induced platelet aggregation, platelet P-selectin expression, and the activity of phospho-phospholipase C beta 3 (P-PLC beta 3) and total-phospholipase C beta 3 (T-PLC beta 3), which play key roles in the signal transduction system of platelet in human. It was found that resveratrol blocked platelet aggregation and platelet P-selectin expression induced by U46619 in a concentration-dependent manner. U-73122 and resveratrol had additive effect in inhibiting platelet aggregation and platelet P-selectin expression. Resveratrol (final concentration was 50 microM) could reduce the ratio of P-PLC beta 3 to T-PLC beta 3. Taken together, these results show that resveratrol suppresses U46619-induced platelet aggregation and P-selectin expression partly through the decrease of the activity of phospholipase C beta of platelets.     

Dipyridamole: pharmacokinetics and effects on aspects of platelet function in man.

1. The effect of dipyridamole on platelet function was measured in twelve normal subjects given 150 or 200 mg tablets as single and multiple doses, and in six subjects given single doses of 25, 50 and 100 mg and multiple doses of 50 mg 8 hourly. 2. Platelet aggregation was measured in response to ADP and collagen. In the subjects given 150/200 mg, the platelets were assayed for content of cyclic AMP and for formation of thromboxane after addition of collagen. The responses to ADP and collagen and the cyclic AMP content were assessed in both the presence and absence of added PGE1. The pharmacokinetics of dipyridamole were studied in all subjects. 3. One hour after 150/200 mg single doses of dipyridamole there was significant inhibition of platelet aggregation in response to both collagen and ADP. There was no detectable effect on aggregation at other time points or with lower doses of dipyridamole. The addition of PGE1 to platelets prior to testing did not enhance the effect of dipyridamole on platelet aggregation. 4. In multiple doses, dipyridamole (150/200 mg twice daily for 11 days) had no detectable effect on platelet aggregation. 5. Dipyridamole did not have any effect on platelet cyclic AMP content, whether or not PGE1 was added prior to assay. 6. Dipyridamole did not affect platelet thromboxane formation. 7. Plasma dipyridamole concentrations were maximal 1-2 h after ingestion, at the same time that inhibition of platelet aggregation was detected. The concentrations declined in a biexponential fashion, with a terminal half life of 24.1 +/- 1.9 h (mean +/- s.e. mean). In six of the 17 subjects, the mean steady state plasma concentration was less than 75% of the value predicted from the single dose data.