微量离心法应用于街头献血者血红蛋白及脂肪血检测

目的探讨微量离心法应用于街头献血者的血红蛋白及脂肪血检测的可行性。方法应用毛细血管专用离心机及全自动血细胞分析仪,分别对600例健康献血者标本进行血细胞比容、血红蛋白浓度及脂肪血检测,检测数据进行统计学处理,界定微量离心法Hct筛查临界值及血浆乳糜程度。结果 Hb与Hct高度正相关,吸取95mm（60μl）末梢血,红细胞压积层≥43.5mm可认为Hb≥120g/L、红细胞压积层≥39mm可认为Hb≥115g/L;脂肪血报废率下降1.4%。结论应用微量离心法同时进行Hct及脂肪血检查,操作简单,无需试剂,仪器小巧便于携带。可用末梢血样,与其他献血前血液检测项目同时采样,不会增加献血者的不适。可替代血站系统目前采用的硫酸铜比重法及献血前咨询来控制血液Hb及乳糜程度检查。     

冻存前滤除白细胞对解冻红细胞质量影响的研究分析

目的探讨冻存前滤除白细胞对冰冻解冻去甘油红细胞的血液质量和生化指标的影响。方法随机采集20名无偿健康献血者全血400mL×20袋,常规分离血浆制备成悬浮红细胞2μ×20袋。根据配对设计方法,将血液等量分成两份1μ×20袋,其中20袋在2~4℃静置2~4h后用一次性白细胞滤器滤除白细胞为滤白组,另20袋不做其他处理为非滤白组。两组红细胞经40%甘油化后冻结,保存在低于-65℃的超低温冰箱至少30d,解冻后红细胞复悬于MAP红细胞保存液中,4℃保存。分别在0、7、14、21d检测解冻红细胞的血液学和生化学指标。结果滤白组冰冻解冻去甘油红细胞较非滤白组冰冻解冻去甘油红细胞的溶血率低、氨浓度低及ATP浓度高,两组比较差异有统计学意义(P0.05)。滤白组可见LDH水平明显抑制,钾浓度增加速度相对较缓。结论滤白组解冻红细胞长期生存能力较非滤白组解冻红细胞强。     

献血者自身冷抗体致白细胞过滤不畅1例

正血液经白细胞过滤器可去除血液成分中99.9%以上的白细胞。但在白细胞过滤时因血液粘稠度高、滤器质量问题、制备过程操作不当等原因经常会出现白细胞过滤不畅现象,但因献血者自身冷抗体导致白细胞过滤不畅的现象并不多见。本站发现1例因献血者血浆中含有冷抗体,导致白细胞过滤不畅,经室温(20℃)复温后过滤顺畅,但离心制备后红细胞溶血报废,现报告如下。1材料与方法1.1资料来源2015年11月26日本中心采集的1袋300     

去白细胞悬浮红细胞的临床应用

目的观察去白细胞前后血液质量变化,及临床输注去白细胞悬浮红细胞降低非溶血性发热性输血反应（FNHTR）的效果。方法随机采集全血30袋（300ml/袋）,在滤除白细胞操作前、后分别留取血液样本进行检测,分别记录过滤前后血液指标。分析临床130例输注去白细胞悬浮红细胞患者和133例输悬浮红细胞患者FNHTR的发生率。结果过滤后红细胞（RBC）回收率大于90%、白细胞去除率99.99%、血小板去除率76.66%、过滤后血液容积减少（44.28±3.12）ml,血红蛋白（HGB）含量、红细胞压积Hct、游离血红蛋白含量FHb、MCV、MCH及MCHC过滤前后的差异无统计学意义（P〉0.05）。去白细胞组FNHTR发生率（2.95%）低于悬浮红细胞组（12.29%）,差异有统计学意义（P〈0.01）。结论国产一次性使用去白细胞滤器可有效去除白细胞,且红细胞回收率大于90%,达到或超过血站基本标准中对去白细胞血液成分的要求。临床输注去白细胞悬浮红细胞能有效减少非溶血性发热性输血反应的发生,提高输血质量。     

不同白细胞滤除方式对去白细胞悬浮红细胞溶血的影响

目的 探讨不同白细胞滤除方法及血液储存时间对红细胞溶血的影响,选择并建立合适的操作方法,降低溶血率.方法 采用采血袋与滤器一体的多联采血联袋.将采集的全血分2个阶段,分别采用直接混匀过滤和少量血浆浸润滤盘法制备去白细胞悬浮红细胞,观察不同制备方法、24h内的血液与储存24 h后的血液过滤后对红细胞溶血的影响.结果 少量血浆浸润滤盘法制备的去白细胞悬浮红细胞溶血比例明显低子直接混匀过滤法;储存24h以内和24h以后过滤的红细胞溶血率有统计学差异.结论 在24 h内采用少量血浆浸润滤盘法能有效降低去白细胞悬浮红细胞因溶血而造成的血液不合格率.     

脂肪血浆去除乳糜微粒前后的对比分析

目的分析脂肪血浆去除乳糜微粒(CM)后的质量变化。方法取30袋脂肪血浆于-50℃冰箱冰冻,使用(3±2)℃解冻冰冻血浆,后低温离心,去除脂肪血浆的CM。结果脂肪血浆去除CM后,外观清晰度明显改善,与正常血浆相当;与去除前比较,三酰甘油水平及总胆固醇水平均降低,差异均有统计学意义(P<0.05);血浆凝血因子FⅧ、FV、纤维蛋白原及血浆总蛋白水平差异均无统计学意义(P>0.05);输血相关传染病标志物检测均合格。结论脂肪血浆去除CM后,不影响血液的安全性,质量符合国家质量标准要求,可用于临床输注。     

鱼精蛋白防止肝素化血浆纤维蛋白凝块析出

鱼精蛋白防止肝素化血浆纤维蛋白凝块析出陈大宁庄一义（南京军区总医院检验中心，南京２１０００２）关键词血液透析肝素化纤维蛋白血液透析病人肝素化血液在分离血浆后，由于肝素逐渐失活，使血浆中纤维蛋白不断析出，形成纤维蛋白微小凝块堵塞自动化分析仪的管道，影响...     

郑州市无偿献血者血液检测不合格原因分析及预防

目的分析郑州市无偿献血者血液检测不合格的原因,探讨其预防措施。方法对2013年1月—2016年12月河南省红十字血液中心采集的821 025份无偿献血血液进行检测,统计不合格率,并进行分析。结果 821 025份血液共3 138 807u,不合格率为5.84%;浓缩血小板、红细胞类、血浆类检验项目不合格率较高,分别为3.80%、2.68%、2.64%;浓缩血小板、血浆类非检验项目不合格率较高,分别为64.51%、7.18%;冷沉淀凝血因子医院退血率较高,为0.21%;检验项目不合格血液中,谷丙转氨酶、乙型肝炎表面抗原不合格率较高,分别为58.46%、15.04%;非检验项目不合格血液中,脂血、破袋或漏液所占比例较高,分别为91.00%、3.19%;医院退血血液中,破袋或漏液、凝块或絮状物所占比例较高,分别为70.45%、19.69%。结论郑州市无偿献血血液不合格较高,与多种因素有关,加强献血前初筛,规范血液采集、分离及运输是降低血液检测不合格率的重要措施。     

全血及成分血质量抽检结果趋势及不合格原因分析

目的对全血及成分血抽检结果进行趋势及不合格分析,为采供血过程提供纠正预防措施提供依据。方法利用Excel对资料数据绘图并进行趋势分析。结果全血血细胞比容最低0.30;悬浮红细胞比容抽检合格率为79.2%,悬浮红细胞比容最低0.42,不同厂家的采血袋悬浮红细胞血比容合格率差异有统计学意义(P0.05)。去白悬浮红细胞前3批白细胞残留量有整体偏高趋势,合格率为50%,后3批合格率为100%;冷血过滤残留白细胞明显低于温血过滤(P0.05)。冷沉淀Ⅷ因子4月份和10月份抽检合格率较低,冬季合格率较高。结论严格执行采血前的Hb筛查工作,在离心制备时尽可能将血浆分离出,提高悬浮红细胞血比容合格率;滤除白细胞热过滤改为冷过滤,残留白细胞合格率明显上升;加强采血及血液制备环境温度的控制,确保Ⅷ因子含量不失活。     

献血者含不规则抗体血液的处置及临床应用研究

目的探讨将含不规则抗体血液制备成洗涤红细胞或者冰冻解冻去甘油红细胞(以下简称冰冻红细胞)使用的可行性,为制定献血者含不规则抗体血液处置原则、节约血液资源、减少血液浪费提供依据。方法献血者血液检出不规则抗体时,确定其类型、效价,将血浆报废,对应的悬浮红细胞按照标准化洗涤方案制备处置,观察不同效价、类型抗体血液的洗涤效果,并追踪临床应用情况。结果 2017年5月～2019年7月,本中心共筛查出45例含不规则抗体血液,经制备处置后,总体合格率为91.1%。IgM类合格率100%(25/25);IgG类合格率80.0%(16/20),其中不合格的4例均为IgG-D,效价在8～16,加洗1～2次后均符合要求。其中23例应用于临床,未发生输血反应。结论含不规则抗体的血液经制备处置后应用于临床是可行的,可大大节约血液资源;不规则抗体为IgG-D且效价较高时,应增加洗涤次数或洗涤后留样检测,无残留抗体时,方能发往临床。     

Correlation of the echogenicity and structure of clotted blood

The echogenicity of clotting human blood in sterile plastic bags was studied for up to 21 days and correlated with the histology of the clots. As observed with a 6-MHz A-scanner and a 5-MHz real-time gray-scale scanner, fresh blood and fresh clots were rich in internal echoes. The blood clot gradually became anechoic as a result of erythrocyte packing, hemolysis, and formation of fibrin thrombus, except for the irregular upper border, which also remained histologically inhomogeneous. This study shows that the echogenicity of a hematoma or clot is dependent on its histology; namely, on the distribution and integrity of cells within the clot.     

在纤维蛋白凝块上诱导脐血间充质干细胞向膀胱尿路上皮细胞的分化

背景:在纤维蛋白凝块上诱导脐血间充质干细胞分化为膀胱尿路上皮细胞,尚无明确方案。目的:探讨脐血来源间充质干细胞在纤维蛋白凝块上分化为膀胱尿路上皮细胞的可能性。方法:实验组将脐血间充质干细胞接种于人工制备的脐血来源的纤维蛋白凝块上与胎儿膀胱尿路上皮细胞共培养,对照组将脐血间充质干细胞接种于人工制备的脐血来源的纤维蛋白凝胶上仅用DMEM/F1培养基培养。结果与结论:实验组的脐血间充质干细胞逐渐伸展呈多角形,细胞扁平变大,透射电子显微镜下具有膀胱尿路上皮细胞的超微结构,抗广谱角蛋白(AE1/AE3)免疫组织化学染色阳性。对照组的脐血间充质干细胞呈长梭形,抗广谱角蛋白(CKAE1/AE3)免疫组织化学染色阴性。提示在纤维蛋白凝块上诱导脐血间充质干细胞有分化为膀胱尿路上皮细胞的能力。     

Plasminogen on the surfaces of fibrin clots prevents adhesion of leukocytes and platelets

Summary. Background and Objectives: Although leukocytes and platelets adhere to fibrin with alacrity in vitro, these cells do not readily accumulate on the surfaces of fibrin clots in vivo. The difference in the capacity of blood cell integrins to adhere to fibrin in vivo and in vitro is striking and implies the existence of a physiologic antiadhesive mechanism. The surfaces of fibrin clots in the circulation are continually exposed to plasma proteins, several of which can bind fibrin and influence cell adhesion. Recently, we have demonstrated that adsorption of soluble fibrinogen on the surface of a fibrin clot results in its deposition as a soft multilayer matrix, which prevents attachment of blood cells. In the present study, we demonstrate that another plasma protein, plasminogen, which is known to accumulate in the superficial layer of fibrin, exerts an antiadhesive effect. Results: After being coated with plasminogen, the surfaces of fibrin clots became essentially non‐adhesive for U937 monocytic cells, blood monocytes, and platelets. The data revealed that activation of fibrin‐bound plasminogen by the plasminogen‐activating system assembled on adherent cells resulted in the generation of plasmin, which decomposed the superficial fibrin layer, resulting in cell detachment under flow. The surfaces generated after the initial cell adhesion remained non‐adhesive for subsequent attachment of leukocytes and platelets. Conclusion: We propose that the limited degradation of fibrin by plasmin generated by adherent cells loosens the fibers on the clot surface, producing a mechanically unstable substrate that is unable to support firm integrin‐mediated cell adhesion.     

Assessing the viscoelastic properties of thrombus using a solid-sphere-based instantaneous force approach

The viscoelastic properties of thrombus play a significant role when the clot closes a leak in a vessel of the blood circulation. The common method used to measure the viscoelastic properties of a clot employs a rheometer but this might be unsuitable due to the clot fiber network being broken up by excessive deformation. This study assessed the feasibility of using a novel acoustic method to assess the viscoelastic properties of blood clots. This method is based on monitoring the motion of a solid sphere in a blood clot induced by an applied instantaneous force. Experiments were performed in which a solid sphere was displaced by a 1 MHz single-element focused transducer, with a 20 MHz single-element focused transducer used to track this displacement. The spatiotemporal behavior of the sphere displacement was used to determine the viscoelastic properties of the clot. The experimental system was calibrated by measuring the viscoelastic modulus of gelatin using different types of solid spheres embedded in the phantoms and, then, the shear modulus and viscosity of porcine blood clots with hematocrits of 0% (plasma), 20% and 40% were assessed. The viscoelastic modulus of each clot sample was also measured directly by a rheometer for comparison. The results showed that the shear modulus increased from 173 ± 52 (mean ± SD) Pa for 40%-hematocrit blood clots to 619.5 ± 80.5 Pa for plasma blood clots, while the viscosity decreased from 0.32 ± 0.07 Pa?s to 0.16 ± 0.06 Pa?s, respectively, which indicated that the concentration of red blood cells and the amount of fibrinogen are the main determinants of the clot viscoelastic properties.     

The role of fibroblasts in organization and degradation of a fibrin clot

Older clots become less sensitive to fibrin degradation than newly formed ones. A possible role for fibroblasts in this defective thrombus lysis was studied. A system has been developed in which different clones of fibroblasts were incorporated into a floating whole blood clot. The effect of the incorporated fibroblasts on clot lysis has been analyzed in relation to their basic characteristics: clot retraction, production of plasminogen activators (PAs) and their inhibitors (PAIs), and secretion of collagen. In neoplastic fibroblast-enriched clots, secretion of PA was associated with spontaneous lysis of a whole blood clot. Normal fibroblasts, secreting levels of PA and PAI similar to those of the cancer cells, did not induce spontaneous lysis of the clot. Moreover, these cells protected whole blood clot from thrombolysis by added PA. Our data show that the resistance to fibrin clot degradation induced by normal fibroblasts was mainly mediated by collagen secretion and deposition rather than PAI secretion or retraction of the clot. We suggest a key role for normal fibroblasts in the acquisition of resistance to proteolytic fibrin degradation of whole blood clots through the secretion of collagen.     

The spatial dynamics of fibrin clot dissolution catalyzed by erythrocyte‐bound vs. free fibrinolytics

Summary. Background: Coupling fibrinolytic plasminogen activators to red blood cells (RBCs) has been proposed as an effective, yet safe method of thromboprophylaxis, because of increased circulation lifetime and reduced propensity to induce hemorrhage by selectivity for nascent thrombi rather than pre‐formed hemostatic clots. Objectives and methods: We used confocal microscopy of fluorescently labeled fibrin and erythrocytes in plasma‐derived clots to study the spatial dynamics of lysis catalyzed by RBC‐coupled vs. free plasminogen activators (RBC‐PA vs. PA). Results: Clot lysis catalyzed by free PA progressed gradually and uniformly. In contrast, distinct holes formed surrounding RBC‐PA while the rest of the clot remained intact until these holes enlarged sufficiently to merge, causing sudden clot dissolution. Compared with naïve RBCs within clots lysed by free PA, RBC‐PA moved faster inside the fibrin network prior to clot dissolution, providing a potential mechanism for spatial propagation of RBC‐PA induced lysis. We also showed the focal nature of fibrinolysis by RBC‐PA as dense loading of PA onto RBCs initiates more efficient lysis than equal amounts of PA spread sparsely over more RBCs. In an in vitro model of clots exposed to buffer flow, incorporated RBC‐PA increased permeability and formed channels eventually triggering clot dissolution, whereas clots containing free PA remained intact. Conclusions: Clot lysis by RBC‐PA begins focally, has a longer lag phase when measured by residual mass than homogeneous lysis by PA, is propagated by RBC‐PA motility and provides more effective clot reperfusion than free PA, making RBC‐PA attractive for short‐term thromboprophylaxis.     

B-mode sonography of blood clots

Systematic evaluation of blood clot echogenicity was performed with five different transducer frequencies in two experiments. In the first experiment, blood clots were insonified at five different time periods; from immediately after clotting up to 96 hours after clotting. In the second experiment, blood clots of four different hematocrits (48 to 20%) and clots of hemolysed blood were insonified. The clots, with normal hematocrits, were highly echogenic when imaged with 5, 7.5 and 10-MHz transducers immediately and 24 hours after clotting. The echo intensity decreased over the following days until it almost disappeared at 96 hours after clotting. Clot echogenicity was not observed with 2.25 and 3.5-MHz transducers, except at the interface between retracted clot and serum. Clot echogenicity decreased in proportion with the hematocrit. Hemolysed blood clots were not echogenic. It is concluded from this study that fresh blood clots are echogenic soon after thrombosis with high resolution imaging and this echogenicity diminishes with time. Ultimately, with organization and lamination, echogenicity will recur.     

Fibrin induces release of von Willebrand factor from endothelial cells.

Addition of fibrinogen to human umbilical vein endothelial cells in culture resulted in release of von Willebrand factor (vWf) from Weibel-Palade bodies that was temporally related to formation of fibrin in the medium. Whereas no release occurred before gelation, the formation of fibrin was associated with disappearance of Weibel-Palade bodies and development of extracellular patches of immunofluorescence typical of vWf release. Release also occurred within 10 min of exposure to preformed fibrin but did not occur after exposure to washed red cells, clot liquor, or structurally different fibrin prepared with reptilase. Metabolically labeled vWf was immunopurified from the medium after release by fibrin and shown to consist of highly processed protein lacking pro-vWf subunits. The contribution of residual thrombin to release stimulated by fibrin was minimized by preparing fibrin clots with nonstimulatory concentrations of thrombin and by inhibiting residual thrombin with hirudin or heating. We conclude that fibrin formed at sites of vessel injury may function as a physiologic secretagogue for endothelial cells causing rapid release of stored vWf.     

Blood components produced from whole blood using the Atreus processing system

BACKGROUND: The Atreus 2C+ system (Gambro BCT) automates whole blood (WB) processing into a single device. This study compared the quality of red blood cells (RBCs), fresh‐frozen plasma (FFP), and buffy coats (BCs) made from WB held with or without active cooling. STUDY DESIGN AND METHODS: WB was collected into Atreus disposables and stored with (n = 20) or without (n = 20) active cooling for 14 to 18 hours at 22 ± 2°C before processing with the Atreus. Two RBC leukodepletion filters were assessed, and markers of RBC quality were tested to Day 42. BCs were held for 3 hours before testing, plasma was tested, and samples were frozen for coagulation analysis. RESULTS: RBCs met UK specifications for volume, hemoglobin content (48 ± 5 g), and hematocrit (Hct). Hemolysis, adenosine triphosphate, 2,3‐diphosphoglycerate, potassium, glucose, and lactate throughout storage were all within expected ranges. No differences were seen in RBC produced from WB held with or without active cooling. FFP units met UK specification for volume, total protein, cellular contamination, and coagulation factors. No differences were seen in FFP produced from WB held with or without active cooling. The Hct of BCs produced from WB held without active cooling was lower than in BCs from WB held with active cooling; no differences in activation were seen. CONCLUSION: From these in vitro data, blood components produced using the Atreus appear suitable for clinical use, with no clinically significant difference in the quality of components from WB held at ambient temperature overnight with or without active cooling.     

悬浮红细胞及血浆输注对大量输血手术患者凝血功能的影响

目的观察悬浮红细胞及血浆输注对大量输血手术患者凝血功能的影响。方法选取2018年12月至2020年5月该院大量输血手术患者63例,输注悬浮红细胞的31例患者为对照组,输注悬浮红细胞及血浆的32例患者为联合组。比较两组输注前后凝血功能指标[凝血酶原时间(PT)、纤维蛋白原(FIB)、凝血酶时间(TT)、活化部分凝血活酶时间(APTT)]、血液流变学指标[血小板计数(PLT)、血红蛋白(Hb)、血细胞比容(HCT)],以及住院时间、凝血功能障碍发生率。结果联合组输注后24、72 h PT、APTT、TT短于对照组,FIB、PLT水平高于对照组(P<0.05)。两组输注前,输注后24、72 h HCT、Hb比较,差异无统计学意义(P>0.05)。两组住院时间、凝血功能障碍发生率比较,差异无统计学意义(P>0.05)。结论悬浮红细胞及血浆输注对大量输血手术患者凝血功能、PLT的影响较小,有助于预防凝血功能障碍的发生,促进预后改善。