升血小板胶囊联合重组人血小板生成素治疗特发性血小板减少性紫癜的疗效观察

目的观察升血小板胶囊联合重组人血小板生成素注射液治疗特发性血小板减少性紫癜(ITP)的临床疗效。方法选取2016年1月—2016年10月在信阳市中心医院住院治疗的ITP患者68例,随机分为对照组和治疗组,每组各34例。对照组皮下注射重组人血小板生成素注射液,300 U/(kg·d),1次/d。治疗组在对照组的基础上口服升血小板胶囊,1.8 g/次,3次/d。两组患者均治疗14 d。评价两组患者临床疗效,同时比较治疗前后两组患者血小板计数和血清相关血小板抗体(PAlgG)升高例数差异。结果治疗后,对照组临床总有效率为70.59%,显著低于治疗组的91.18%,两组比较差异具有统计学意义(P0.05)。治疗后,两组血小板计数均明显升高,同组比较差异具有统计学意义(P0.05);且治疗组患者血小板计数水平显著高于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,治疗组血清PAIgG水平大于108 ng/10PA的例数显著少于对照组,两组比较差异具有统计学意义(P0.05)。结论血小板胶囊联合重组人血小板生成素能显著缩短ITP患者血小板恢复的时间,提高临床效果,具有一定的临床推广应用价值。     

发热 贫血 血小板减少 神经系统异常

患儿男,13岁7个月。2010-11-04以发热伴面色苍白1周为主诉入住中国医科大学附属第一医院儿科病房。患儿1周前开始无诱因发热伴颈部包块,无触痛,咳嗽无痰,体温最高达39℃,同时伴有面色苍白、乏力,无皮肤黄染,尿色略黄。曾于当地医院就诊,静滴抗炎类药物3d(具体药物不详),病情无好转,化验发现贫血严重。患儿病来时有烦躁,无水肿,睡眠可,精神差,进食差,鼻衄2次,黑便1次,无明显腹痛及尿量减少。     

特发性血小板减少性紫癜患者脾脏CD5+和CD5-B细胞共同参与血小板自身抗体的产生

目的：研究慢性特发性血小板减少性紫癜（ITP）患者脾脏CD5^ B细胞水平的变化及CD5^ 和CD5^-B细胞与血小板膜糖蛋白（GP）特异性自身抗体产生的关系，以识别致病B细胞亚群。方法：应用双色流式细胞仪检测8例慢性ITP患者脾脏CD5^ B细胞水平。选择4例血浆抗GPⅡb/Ⅲa和抗GP Ⅰb/Ⅸ抗体双阳性ITP切脾患者，应用Ficoll密度梯度离心及花环形成分离法分离脾脏B淋巴细胞，继而采用镝产珠分选法分选、纯化CD5^ B细胞和CD5^-B细胞，并分别进行体外培养，应用改良MAIPA法检测血浆和细胞培养上清液的血小板特异性抗体。结果：ITP患者脾脏CD5^ B细胞水平圈晨自身免疫性疾病患者略有增高，二者之间差异无统计学意义。CD5^ B细胞水平与患者血小板计数无相关性。4例血浆抗GPⅡb/Ⅲa抗体和抗GPⅠb/Ⅸ抗体双阳性。另外1例CD5^ B细胞培养液抗GPⅡb/Ⅲa抗体阴性，抗GPⅠb/Ⅸ抗体阳性;CD5^-B细胞培养液抗GPⅡb/Ⅲa抗体和抗GPⅠb/Ⅸ抗体双阳性。结论：脾脏CD5^ 和CD5^-B细胞 均可产生血小板GP特异性自身抗体，抗体产生种类和滴度无明显差异。提示二者共同参与了ITP的发病过程。     

自身免疫性血小板减少性紫癜相关抗体的研究

目的　探索对自身免疫性血小板减少性紫癜 (AITP)诊断特异和敏感的实验方法。方法　采用单克隆抗体特异性俘获血小板抗原技术 (MAIPA技术 )并加以改进 ,对比检测血小板洗脱液和血浆中血小板膜糖蛋白特异性抗体。结果　AITP患者血浆游离抗血小板膜糖蛋白 (GPⅡb Ⅲa、GPⅠb Ⅸ )抗体总阳性率为 38.89%(5 4例中 2 1例 ) ,洗脱血小板表面抗血小板膜糖蛋白 (GPⅡb Ⅲa、GPⅠb Ⅸ )抗体总阳性率为 6 8.5 2 %(5 4例中 37例 ) ,两者差异有显著性 (校正 χ2 =19.39,P <0 .0 0 5 )。原发AITP组血浆游离及洗脱血小板表面抗血小板糖蛋白 (GPⅡb Ⅲa、GPⅠb Ⅸ )特异性抗体总阳性率与继发性AITP组比较差异无显著性。AITP患者血小板数量与自身抗体滴度呈明显负相关。结论　MAIPA法检测血小板洗脱液中抗血小板糖蛋白抗体在AITP的诊断和治疗中具有高度特异性 ,且敏感性较血浆抗体检测显著提高。     

流式免疫微球芯片技术分析特异性血小板自身抗体

目的建立流式免疫微球芯片技术（FCIBA）分析特异性血小板自身抗体的新方法。方法使用不同红色荧光强度的微球，包被不同血小板膜蛋白单克隆抗体（抗GPⅡb／Ⅲa、抗GPⅠa／Ⅱa、抗GPⅣ、抗GPⅠb／Ⅸ和抗HLA-ABC单克隆抗体），制备特异性血小板自身抗体检测微球，用流式细胞仪检测微球捕获的血小板抗原-自身抗体复合物，检测结果与微孔板改进抗原捕获ELISA（MACE）进行比较，并检测了部分免疫性以及非免疫性血小板减少性紫癜患者血清中的5种特异性血小板自身抗体。结果FCIBA分析抗GPⅡb／Ⅲa、抗GPⅠa／Ⅱa、抗GPⅣ、抗GPⅠb／Ⅸ和抗HLA-ABC5种特异性血小板自身抗体，批内重复性变异系数（CV）分别为4．82％、6．09％、5．04％、5．73％、5．30％；稀释试验呈对数线性相关，相关系数（r）分别为0．9972．0．9966．0．9988．0．9965、0．9982；新方法对5种特异性血小板自身抗体的分析结果均与MACE试验结果高度相关，r值分别为0．9289、0．9224、0．8894、0．9100、0．9134（P均〈0．01）。新方法分析98例免疫性血小板减少性紫癜患者的血清样本，血小板特异性自身抗体检出的阳性率为51．69％；其中，抗GPⅡb／Ⅲa为40．82％，抗GPⅠa／，Ⅱa为24．45％，抗GPⅣ为19．39％，抗GPⅠb／Ⅸ为32．65％，抗HLA-ABC为17．35％。新方法分析40例非免疫性血小板减少性紫癜患者的血清样本，阳性率为0。结论本研究建立了FCIBA分析特异性血小板自身抗体的新方法，可同时分析抗血小板GPⅡb／Ⅲa、GPⅠa／，Ⅱa、GPⅣ、GPⅠb／Ⅸ和HLA-ABC5种特异性血小板自身抗体。     

血栓性血小板减少性紫癜患者vWF裂解蛋白酶抗原和活性的检测及意义

目的研究血管性血友病因子裂解蛋白酶(ADAMTS13)抗原含量和活性在血栓性血小板减少性紫癜(TTP)患者及遗传性 TTP 家族突变携带者中变化的情况。方法用残余胶原结合实验(RCBA)检测13例 TTP 患者共28份血浆标本[含血浆置换(PE)前后]及10例携带者的 ADAMTS13活性;用新近建立的三抗体夹心酶联免疫反应法检测标本的 ADAMTS13抗原含量。结果正常对照组 ADAMTS13含量为(600.93±145.36)mU/ml(设白种人混合血浆的 ADAMTS13抗原含量为1000mU/ml),活性为(74.79±11.81)%。遗传性 TTP 患者 ADAMTS13抗原含量和活性治疗前和发病间期均明显减低,PE 后恢复;其家族中携带者 ADAMTS13抗原含量为(331.40±109.85)mU/ml,活性为(66.79±12.82)%(与对照组比较,P 值分别<0.01和>0.05);原发性 TTP 患者 PE 前 ADAMTS13抗原含量为(98.7±82.08)mU/ml,活性为(22.23±19.07)%(与对照组比较,P 值均<0.01);PE 后ADAMTS13 抗原含量为(449.4±232.33)mU/ml,活性为(60.92±22.33)%(与对照组比较,P 值分别<0.01和>0.05);1例继发性 TTP 患者 PE 后 ADAMTS13抗原含量远高于正常,活性仅为6.00%结论治疗前的 TTP 患者 ADAMTS13抗原含量和活性均明显减低。大多数患者两指标变化趋势一致,也有个别患者两指标变化趋势相反,前者可能因为遗传因素或体内免疫系统的廓清作用,后者可能因为抗 ADAMTS13抗体仅抑制了 ADAMTS13的活性而未影响其抗原的含量或其他未知原因所致。     

A rapid test for the diagnosis of thrombotic thrombocytopenic purpura using surface enhanced laser desorption/ionization time-of-flight (SELDI-TOF)-mass spectrometry

BACKGROUND: Thrombotic thrombocytopenic purpura (TTP), a life-threatening thrombotic microangiopathy, requires immediate diagnosis and plasma exchange therapy. Development of TTP is related to functional deficiency of ADAMTS-13 protease that leads to the accumulation of ultra large von Willebrand factor (VWF) and subsequent platelet thrombosis. Currently no clinical test is available for the rapid detection of ADAMTS-13 activity. OBJECTIVES: The goal is to devise a novel method to rapidly detect functional activity of ADAMTS-13 and improve clinical outcome. METHODS AND RESULTS: A recombinant VWF substrate containing the ADAMTS-13 cleavage site and a 6X Histidine tag was cleaved by ADAMTS-13 in a dose-dependent manner, generating approximately 7739 Da peptide containing a 6X Histidine tag. This cleaved peptide, bound to an IMAC/Nickel ProteinChip, was quantified using Surface Enhanced Laser Desorption/Ionization Time-of-flight Mass Spectrometry (SELDI-TOF-MS). The assay is capable of quantifying ADAMTS-13 activity as low as 2.5% in plasma within 4 h. When the cleaved peptide was quantified as a ratio of an internal control peptide, the test displayed good reproducibility, with an average inter-assay coefficient of variation (CV) of < 33%. Further validation revealed a mean ADAMTS-13 activity of 92.5% +/- 16.6% in 39 healthy donors. Sixteen patients with idiopathic TTP displayed mean ADAMTS-13 activity of 1.73% +/- 3.62%. Further utility of this novel method includes determining the inhibitory titer of ADAMTS-13 antibody in cases of acquired TTP. CONCLUSIONS: We have devised a novel SELDI-TOF-MS assay that offers a rapid, cost-effective, and functionally relevant test for timely diagnosis and management of TTP.     

急性ITP患儿外周PPAR-γ mRNA表达变化及与IL-18水平的相关性

目的 检测急性特发性血小板减少性紫癜(idiopathic thrombocytopenic purpura,ITP)惠儿外周血浆中IL-18对血淋巴细胞PPAR-γ mRNA表达变化影响.方法 序贯收集符合试验入选标准的急性ITP患儿53例,同时收集相匹配的同期体检儿童50例作为对照.将血液标本平均分为5份,在取样1、24、48、72 h后,分别采用RT-PCR法检测ITP患儿外周血淋巴细胞PPAR-γ mRNA的表达,蛋白免疫印迹(western blot)检测PPAR-γ蛋白的含量,ELISA法检测血浆中IL-18水平.结果 急性ITP组白细胞表达PPAR-γ蛋白在1、24、48、72 h与对照组PPAR-γ蛋白分别相比均明显增强,差异有显著性(t_1=7.52,t_(24)=16.38,t_(48)=9.65,t_(72)=12.34,均P<0.05);急性ITP患儿在不同的时间点测定的PPAR-γ mRNA表达水平明显均高于正常对照组(t_1=9.25,t_(24)=14.24,t_(48):8.69,t_(72)=16.14,均P<0.05),同样发现急性ITP患儿血浆IL-18水平均显低于正常对照组(t_1=6.38,t_(24):9.65,t_(48)=8.91,t_(72)=7.19,均P<0.05);血浆IL-18水平与PPAR-γ mRNA表达呈负相关(r=-0.89,P<0.05).结论 血浆中IL-18生成量的减少可能导致了急性ITP患儿外周血淋巴细胞PPAR-γ表达的增加,调控Th1/Th2细胞分化功能出现障碍,使有效的免疫抑制作用降低,导致自身反应性T细胞激活增多、凋亡减少,使得浆细胞产生抗血小板抗体增多,促进了血小板的破坏.     

丙戊酸钠致血小板减少性紫癜8例临床分析

目的探讨丙戊酸钠致血小板减少性紫癜可能发生机制.方法对8例患儿服用丙戊酸钠在(3～5.5)个月内相继全身出现紫癜样皮诊进行血小板计数,出血时间和药物血浓度监测.结果服用丙戊酸钠3～5.5个月全身出现紫癜样皮疹患儿血小板计数明显降低,出血时间延长,根据本组临床资料显示,血小板减少数量与丙戊酸纳药物血浓度正相关.结论丙戊酸钠能影响患儿的血小板计数和功能.导致血小板计数降低,出血时间延长,患儿以双下肢为主、全身皮肤出现紫癜样皮疹,其中2例并伴有牙龈出血,影响的程度可能与丙戊酸钠的剂量相关.     

Retrospective evaluation of children with immune thrombocytopenic purpura and factors contributing to chronicity

ObjectiveImmune thrombocytopenic purpura (ITP) is the most common cause of acquired thrombocytopenia children. The aim of this retrospective study is to describe presenting features and clinical characteristics of ITP and evaluate clinical course, treatment modalities, and complications and determine the effects of preceding infection history, age, gender, treatment modality, and admission platelet count on chronicity.MethodTwo hundred and eleven patients who were diagnosed ITP and followed-up in Department of Pediatric Hematology, Ankara Children Hematology Oncology Education and Research Hospital between January 2008 and September 2012 were included. Age of the patients, gender, date of admission, date of diagnosis, complaint in the application, previous infection and laboratory tests were recorded.ResultsMean age of the patients on diagnosis was 5.4 ± 4.1 years. The female/male ratio was 1.03. The clinical courses were determined as acute or chronic in 72% and 28% of patients respectively. Mean age at diagnosis was significantly higher in chronic ITP (p < 0.01). Chronic course was significantly higher in female patients (p < 0.05). The most frequent complaint was bruises on the skin (68%). The most common physical examination findings were petechiae, purpura and ecchymosis (89%). Patients with a history of past infection (53.6%) and who had serologically positive infection (15.6%) frequently had acute course (p < 0.01). The most common serologically positive infection was Rubella. The mean platelet count was significantly higher in chronic ITP (p < 0.01). In the initial treatment of patients admitted in the acute phase, megadose methylprednisolone (MDMP) was used in 31% of patients, intravenous immune globulin (IVIG) in 55% of patients and anti-D in 2% of patients while 12% did not receive any treatment. There were no significant differences between the recurrence rate and treatment modality (p > 0.05).ConclusionIn our study, in females and in patients without any history of past infection, platelet count >20 × 10⁹/L and initial diagnosis age > 10 years were found to increase the probability of chronic disease, which is compatible with the literature.