Frozen Red Blood Cells in Transfusion

Red blood cells (RBCs) can be cryopreserved with shelf life of 10 years. However, shelf life of deglycerolized RBCs in conventional open system is just 24 hours, resulting in sporadic use of Frozen RBC (FS-RBC). Recently Naval Blood Research Laboratory (NBRL) method using ACP 215 (ACP^™ 215 Haemonetics Cell Processing System) has been introduced, where shelf life of deglycerolized RBC is 14 days. FS-RBC unit is prepared from single blood donation, which needs to be glycerolized and deglycerolized. NBRL method using ACP 215 in FS-RBC is described. Deglycerolized unit weighed between 325–350 gm with haemoglobin of 15–18 gm/dl and freeze- thaw- wash RBC recovery of 87%. Transfusion of deglycerolized RBC offered advantages such as elimination of need of crossmatching in emergent situations and reduction of transfusion reactions. FS-RBC by NBRL method using ACP 215 has advantages such as long shelf life, meeting unexpected high blood demand in mass casualties situations or availability of rare blood group requirement of individual patient. FS-RBC can be a potential candidate for Indian Armed Forces Blood programme for uninterrupted blood supply during peace and war.Key Words: Frozen red blood cell, Cryopreservation, Blood transfusion, Armed forces blood programme     

Comparative study of automated cryopreservation of red blood cells

Background

Strategic blood reserves are an important component in meeting blood needs and this can be accomplished through the establishment of a frozen blood program.

Methods

One hundred units of packed RBC were glycerolized using the Haemonetics ACP 215 automated cell processor and placed in a −86 °C deep freezer for freezing and storage. Product weight, hematocrit, RBC count, WBC count and hemoglobin were recorded prior to freezing. Twenty five bags were thawed and deglycerolized after every three months starting at one year from the date of first glycerolization In addition to the earlier parameters the bags were assessed for supernatant osmolality, pH, supernatant hemoglobin, ATP levels and supernatant potassium and from these red cell recovery, percentage hemolysis, supernatant glycerol and red cell viability were estimated. All tests were repeated at the end of 7 and 14 days.

Results

The mean red cell recovery was found to be 86.12% on Day 0 and 84% on Day 14. All the bags showed residual glycerol and pH within the acceptable limits upto Day 14. Percentage hemolysis, Mean ATP levels and mean supernatant potassium levels were within acceptable limits upto Day 14. All the units were sterile upto Day 14.

Conclusion

The data in this study showed that the red cells which were glycerolized using the automated platform ACP 215, frozen at −80 °C for more than a year and deglycerolized again using the ACP 215 had excellent viability while being stored at 4 °C during the 14 days of post-thaw storage.     

RH（一）红细胞洗涤去干甘油的制备过程及体会

随着临床成分输血的广泛发展,以及卫生部《临床输血技术规范》的实施,临床上对RH（一）血的需求量也在增加,但因无偿献血人群RH（一）比例偏低。亚洲人中不足10％,4℃保存的时间短,一方面满足不了临床应用,另一方面极易造成过期作废,为了更好地满足临床对RH（一）血的需求,甘油化红细胞-80℃储存方法是目前长期保存RH（一）红细胞的最佳保存方法。     

Comparative study of automated cryopreservation of red blood cells

Background

Strategic blood reserves are an important component in meeting blood needs and this can be accomplished through the establishment of a frozen blood program.

Methods

One hundred units of packed RBC were glycerolized using the Haemonetics ACP 215 automated cell processor and placed in a −86 °C deep freezer for freezing and storage. Product weight, hematocrit, RBC count, WBC count and hemoglobin were recorded prior to freezing. Twenty five bags were thawed and deglycerolized after every three months starting at one year from the date of first glycerolization In addition to the earlier parameters the bags were assessed for supernatant osmolality, pH, supernatant hemoglobin, ATP levels and supernatant potassium and from these red cell recovery, percentage hemolysis, supernatant glycerol and red cell viability were estimated. All tests were repeated at the end of 7 and 14 days.

Results

The mean red cell recovery was found to be 86.12% on Day 0 and 84% on Day 14. All the bags showed residual glycerol and pH within the acceptable limits upto Day 14. Percentage hemolysis, Mean ATP levels and mean supernatant potassium levels were within acceptable limits upto Day 14. All the units were sterile upto Day 14.

Conclusion

The data in this study showed that the red cells which were glycerolized using the automated platform ACP 215, frozen at −80 °C for more than a year and deglycerolized again using the ACP 215 had excellent viability while being stored at 4 °C during the 14 days of post-thaw storage.     

基于红细胞的载药系统研究进展

基于红细胞(RBC)的新型药物输送系统近年来倍受关注,天然红细胞是机体固有成分,与传统药物载体相比,具有生物相容性高、体内半衰期长等优势。本文综述了红细胞药物载体的特点、制备方法及其载药的最新研究进展;同时,介绍了近年来出现的新型红细胞膜药物输送系统即红细胞膜包裹纳米粒(RBC-NP)和红细胞膜纳米海绵技术。     

Cyclic arginine-glycine-aspartic acid-modified red blood cells for drug delivery: Synthesis and in vitro evaluation

Red blood cells (RBCs) are an excellent choice for cell preparation research because of their biocompatibility, high drug loading, and long half-life. In this study, doxorubicin (DOX) was encapsulated with RBCs as the carrier. The biotin-avidin system binding principle was used to modify biotinylated cyclic arginine-glycine-aspartic acid (cRGD) onto RBC surfaces for accurate targeting, high drug loading, and sustained drug release. The RBC drug delivery system (DDS) was characterized, and the concentration of surface sulfur in the energy spectrum was 6.330%. The physical and chemical properties of RBC DDS were as follows: drug content, 0.857 mg/mL; particle size, 3339 nm; potential value, −12.5 mV; and cumulative release rate, 81.35%. There was no significant change in RBC morphology for up to seven days. The results of the targeting and cytotoxicity studies of RBC DDS showed that many RBCs covered the surfaces of U251 cells, and the fluorescence intensity was higher than that of MCF-7 cells. The IC₅₀ value of unmodified drug-loaded RBCs was 2.5 times higher than that of targeted modified drug-loaded RBCs, indicating that the targeting of cancer cells produced satisfactory inhibition. This study confirms that the RBC DDS has the characteristics of accurate targeting, high drug loading, and slow drug release, which increases its likelihood of becoming a clinical cancer treatment in the future.     

冰冻解冻去甘油红细胞的制备方法及效果评价

目的用ACP215全自动细胞洗涤仪制备冰冻红细胞和冰冻解冻去甘油红细胞,观察冰冻解冻去甘油红细胞的质控指标,评价制备方法和制备效果。方法随机取2-6℃保存第6天的红细胞悬液,用ACP215甘油化红细胞悬液,-65℃以下深低温保存后取出,快速放入38℃水浴箱中融化,再用ACP215洗涤去甘油制备冰冻解冻去甘油红细胞。结果冰冻解冻去甘油红细胞平均红细胞回收率（84.06±2.07）,游离血红蛋白浓度（0.49±0.11）g/L,体外溶血实验（11.59±2.69）%,甘油残余量（3.51±1.27）g/L,残留白细胞（0.48±0.22）%,残留血小板（0.37±0.16）%,细菌培养阴性,满足国标《全血及成分血质量标准》的要求。结论 ACP215制备冰冻红细胞,低温保存并洗涤去甘油,操作方法标准化,快速简便,产品质量稳定可靠,可用于稀有血型及自体储血的保存,缓解供血紧张,保证临床输血的安全性。     

The effects of red blood cell infusion on 10-km race time

The purpose of this study was to investigate the effect of infusion of 400 mL of red blood cells (RBCs) on 10-km track race time, submaximal heart rate, hematocrit, 2,3-diphosphoglycerate, and partial pressure of oxygen at 50% hemoglobin saturation. Six highly trained, male, distance runners twice donated a unit of RBCs, which was frozen for subsequent reinfusion. Eleven weeks after the second donation, they undertook a series of three competitive 10-km races on a standard 400-m track: before infusion, after 100 mL of saline solution, and after 400 mL of autologous, previously frozen deglycerolized RBCs. All subjects took all trials in this double-blind, placebo, crossover, experimental design. Running time was recorded at each 400-m split, and blood was collected prior to each trial. The data were analyzed by analysis of variance. Results following the RBC infusion showed a significantly higher hematocrit concentration, a significantly faster 10-km run, a nonsignificant decrease in submaximal heart rate (10 beats faster 10-km run, a nonsignificant decrease in submaximal heart rate (10 beats per minute), and no significant changes in either 2,3-diphosphoglycerate or partial pressure of oxygen at 50% hemoglobin saturation. Erythrocythemia induced by the infusion of 400 mL of autologous packed RBCs effectively increased performance capacity in a 10-km track race, probably due to an increase in oxygen delivery to the working muscles.     

CELL-DYN1700型自动血细胞计数仪临床应用评价

目的: 对CELL-DYN1700型全自动血球计数分析仪的性能进行临床应用评价。方法: 应用临床标本测定该仪器的精密度、准确度、携带污染率、线性范围。结果: 各项目的批内CV值:白细胞(WBC)1.9%,红细胞(RBC)1.65%,血红蛋白(Hb)1.35%,红细胞比积(HCT)2.0%,平均红细胞体积(MCV)0.93%,血小板计数(BPC)3.2%。批间CV值:高值WBC 2.7%,RBC 2.21%,Hb 1.86%,HCT 2.8%,MCV 1.9%,BPC 6.0%;正常值:WBC 2.1%,RBC 1.9%,Hb 1.7%,HCT 2.3,MCV1.8%,BPC 3.6%;低值WBC 2.0%,RBC 2.1%,Hb 1.68%,HCT 2.4%,MCV 1.92%,BPC 6.8%。线性范围较宽WBC(0.5~90.0)×109/L,RBC(1.0~7.0)×1012/L,Hb(25.0~200.0) g/L、BPC(10.0~999.0)×109/L;携带污染率低:WBC 0.1%,RBC 0%,Hb 0%,BPC 0.5%;准确度高,EDTAK₂抗凝的静脉血仪器法和常规手工法结果比较差异均无显著性(P>0.05)。结论: 该仪器结果准确,重复性好,操作易于掌握,检测速度快,交叉污染小,且易于保养。静脉血、末梢血均能检测18项参数。     

Evaluation of red blood cell transfusion practices with the use of preset criteria.

OBJECTIVE: To assess current red blood cell (RBC) transfusion practices and to determine the potential impact of implementing recently published guidelines on RBC transfusion from the American College of Physicians (ACP). DESIGN: Medical chart review. SETTING: A 219-bed teaching hospital in Kingston, Ont. PARTICIPANTS: All patients over 12 years of age who received RBC transfusions in March 1992. MAIN OUTCOME MEASURES: Need for transfusion according to the ACP guidelines and the number of blood units ordered for each transfusion. RESULTS: A total of 55 patients received 170 RBC units. According to the ACP guidelines 94 (55.3%) of the units were judged unnecessary. The departments of Surgery and Internal Medicine did not differ significantly in the number of unnecessary units (56.4% v. 52.8%). Among the surgical subspecialties, unnecessary transfusion was most common in the orthopedics service (73.5%, p < 0.05). Blood was most frequently ordered 2 units at a time (51.8% of units). Transfusion in normovolemic, hemodynamically stable patients with anemia and unnecessary multiple-unit transfusions were the most common violations of the ACP guidelines. CONCLUSIONS: According to the ACP guidelines, there was significant unnecessary blood use in the hospital surveyed. The guidelines provide a useful framework for assessing transfusion practices but may require further refinement to apply to a broader spectrum of clinical settings.     

4 148 例输血患者红细胞输注无效的调查分析

目的 对临床红细胞输注无效患者进行回顾性分析,并就其影响因素提出相应的临床对策。 方法 选取2015 年5 月-2016 年5 月在湘雅医院行红细胞输注的4 148 例患者,患者输注红细胞后24 h 内复 查血红蛋白（Hb）,与输血前比较,若Hb 未达到预期值,则视为红细胞无效输注,并对调查结果及相关因素 进行统计学分析。结果 医院总无效输血率为12.01%,血液内科无效输血率最高为21.25%,差异有统计学意 义（P <0.05）。红细胞输注效果与性别、年龄、输血次数有关,女性患者无效输血率高于男性患者（P <0.05）; 输血次数越多,无效输血率越高（P <0.05）;患者年龄越大,无效输血率越高（P <0.05）;血液病患者发生无 效输血率高于非血液病患者（P <0.05）。结论 医务人员应重视患者红细胞输血效果,及时采取相应措施,确 保临床输血疗效,做到合理使用血液资源。     

Experimental study on therapy of ultraviolet blood irradiation and oxygenation in acute soman intoxication in rabbits

Objective： To study the therapy effect of ultraviolet blood irradiation and oxygenation （UBIO） on blood AChe activity and lung injury due to acute soman intoxication in rabbits. Methods ：Forty rabbits were randomly divided into 4 groups： normal control group, intoxication group, routine therapy group and UBIO therapy group. Blood AChe activity and artery blood gas were analyzed 2 h after intoxication. ACP and AKP activities in BALF were determined respectively. Results： Blood AChe activity in intoxication group was lower than that in normal control group （P〈0.05）. BALF ACP and AKP activities in intoxication group were higher than that in normal control group. Blood AChe activities in UBIO therapy group increased and were higher than that in intoxication and routine therapy groups. Compared with intoxication group, BALF ACP and AKP activities were decreased （P〈0. 05） in UBIO therapy group, while artery blood pH, PaO2 and SaO2 increased （P〈0. 05）. Conclusion.. UBIO therapy can elevate blood AChe activity and alleviate lung injury induced by soman intoxication. So it may be a new way to treat acute soman intoxication.     

Cyclic arginine-glycine-aspartic acid-modified red blood cells for drug delivery: Synthesis and in vitro evaluation

Red blood cells (RBCs) are an excellent choice for cell preparation research because of their biocompatibility, high drug loading, and long half-life. In this study, doxorubicin (DOX) was encapsulated with RBCs as the carrier. The biotin-avidin system binding principle was used to modify biotinylated cyclic arginine-glycine-aspartic acid (cRGD) onto RBC surfaces for accurate targeting, high drug loading, and sustained drug release. The RBC drug delivery system (DDS) was characterized, and the concentration of surface sulfur in the energy spectrum was 6.330%. The physical and chemical properties of RBC DDS were as follows: drug content, 0.857 mg/mL; particle size, 3339 nm; potential value, −12.5 mV; and cumulative release rate, 81.35%. There was no significant change in RBC morphology for up to seven days. The results of the targeting and cytotoxicity studies of RBC DDS showed that many RBCs covered the surfaces of U251 cells, and the fluorescence intensity was higher than that of MCF-7 cells. The IC₅₀ value of unmodified drug-loaded RBCs was 2.5 times higher than that of targeted modified drug-loaded RBCs, indicating that the targeting of cancer cells produced satisfactory inhibition. This study confirms that the RBC DDS has the characteristics of accurate targeting, high drug loading, and slow drug release, which increases its likelihood of becoming a clinical cancer treatment in the future.     

红细胞直方图和几种贫血相关性的探讨

目的：探讨红细胞直方图的变化和几种贫血的关系。方法：根据sysmex KX—21全自动血细胞分析仪给出的红细胞直方图的变化，观察红细胞大小、分布情况。结果：正常红细胞和各类贫血的红细胞在直方图中均有特定的位置。结论：根据红细胞直方图的变化可以对贫血的类型进行初筛，同时对治疗效果的临床观察有一定的价值。     

国产血库型白细胞滤器在储存添加剂红细胞过滤中的实验研究与临床应用效果评价

目的　探讨国产库型白细胞滤器在储存添加剂红细胞过滤效果。方法　使用国产血库型白细胞滤器 (BC·L 2 0 0 -1型 )过滤 15袋 10天以内的储存添加剂红细胞 (去白RBC) ,检测过滤前后WBC、RBC、PLT的浓度 ,并求出WBC、PLT的去除率 ,WBC残留量及RBC回收率 ;比较过滤前后RBC渗透脆性。并将去白RBC和非去白RBC分别用于两组患者 ,每组患者 2 0 3人 ,观察去白组与非去白组非溶血性发热性输血反应 (NHFTR)的发生率。结果　平均白细胞去除率、血小扳去除率及红细胞回收率分别为 99.3 9%、97.42 %及 93 .5 1%。开始溶血点 :过滤前后比较的t检验得P1 >0 .5 ,完全溶血点 :过滤前后比较的t检验得 0 .2 0 >P2 >0 .10。过滤前后红细胞渗透脆性无显著差异 (P1 >0 .0 5 ;P2 >0 .0 5 )。去白组NHFTR发生率为 1.48% ;非去白组NHFTR发生率为 12 .3 2 % (P <0 .0 0 5 )。结论　该型白细胞滤器的性能完全可以满足临床少白RBC输血的要求。使用该种去白RBC可以大大降低输血不良反应。     

血型对正常成人红细胞血红蛋白释放的影响

目的：观察不同血型（ABO血型系统）对正常成人红细胞血红蛋白释放的影响。方法：利用红细胞血红蛋白释放试验（HRT）,研究比较不同血型的正常成人血红蛋白释放情况。结果：不同血型的正常成人血红蛋白释放量基本相同。结论：血型糖蛋白对红细胞血红蛋白释放没有明显的影响。     

hu-RBC-SCID小鼠模型的建立及野鼠Babesia microti样原虫感染人红细胞的研究

目的 用人红细胞(RBC)替换SCID小鼠自身血循环红细胞而建立的人化小鼠模型．即hu-RBC-SCID模式小鼠，分离并测试野鼠Babesia microti(B．microti)样原虫对人红细胞的感染，以确定人巴贝斯原虫的感染来源。方法 用人O型红细胞注入先期去除自身血循环红细胞的NOD／shi-scid小鼠血循环中，辅以肌内注射兔抗小鼠血红蛋白生成素血清和大鼠抗小鼠RBC单克隆抗体，建成hu-RBC-SCID模式小鼠，接种野鼠感染B．microti的RBC。结果 hu-RBC-SCID模式小鼠的人RBC中，在接种感染B．microti野鼠的RBC后，出现巴贝斯样原虫，并大量增殖。RDNA序列比较发现两类为B.microti样原虫，一类与人的一致。结论 感染野鼠的两类B．microti样原虫均对人RBC具有高度感染性；hu-RBC-SCID模式小鼠的感染，直接提供了人巴贝斯原虫的感染来自野鼠的证据。     

The changes of red blood cell viscoelasticity and sports anemia in male 24-hr ultra-marathoners

Background

In endurance sports, stress, dehydration and release of chemical factors have been associated with red blood cell (RBC) alterations of structure and function, which may contribute to sports anemia, a well-observed phenomenon during long-distance running. Until now, the investigation of the changes of viscoelastic properties of RBC membrane, a decisive factor of RBC deformability to avoid hemolysis, is lacking, especially in an Oriental population.

急性冠脉综合征发生与外周血红细胞源微粒致血栓活性的关系研究

目的 通过评估红细胞源微粒（RMPs）和急性冠脉综合征（ACS）发病之间的关联,描述RMPs在急性冠状动脉血栓形成过程中发挥的作用。方法 研究对象分ACS组（n=200）,其分为急性心肌梗死（AMI,n=100）和不稳定型心绞痛组（UAP,n=100）,对照组为非冠心病组（n=100）。获得患者知情同意后,收集ACS组以及对照组的外周血,离心后加入钙离子载体A23187,得到MPs。用特异性红细胞微粒抗体（glycophorine A）标记RMPs,后经流式细胞仪定量分析。在体外,将ACS患者血浆中提取的RMPs加入对照组血浆中,在荧光标记的血栓底物作用下诱导血栓形成并用荧光光度仪检测RMPs的致血栓活性。结果 RMPs水平在ACS的AMI亚组（163×10²）和UAP亚组（96×10²）都明显高于对照组（12×10²,P<0.05）。ACS患者在每一个时间段相对荧光单位较对照组显著提高（P<0.05）。研究结果显示,高浓度的MPs更易导致血栓形成且所需时间较短,差异有统计学意义（P<0.05）。结论 ACS组的外周血中RMPs水平明显升高,提示RMPs参与了ACS的发生、发展过程。MPs诱导的血栓形成峰值前移,提示RMPs可能诱导凝血酶形成,揭示ACS发生急性血栓与RMPs有关。     

恶性肿瘤患者化疗前后血型抗原和抗体分析

目的　了解恶性肿瘤患者化疗对红细胞血型抗原和抗体的变化。方法　检测 1 0 4例患者化疗前后ABO血型抗原效价、抗 A、抗 B抗体效价 ,A型和O型患者化疗前后抗原抗体效价与积分做了比较 ;抗原检测法 ,将标准血清做倍比稀释加患者的红细胞 ,离心肉眼观察 ,不凝集的前 1管为抗原效价 ,同时以健康人的红细胞做对照 ;抗体检测方法 ,将A型者的血清做倍比稀释后加对应的红细胞 ,观察结果同抗原方法。结果　O型抗 A、抗 B效价与积分无差异 ,A型血型抗原化疗前后有显著性差异 ,化疗后明显低于化疗前 ,P <0 .0 5。O型、A型、B型男女患者抗原抗体效价与积分进行比较 ,O型与B型男女抗原抗体、效价与积分无差异 ;A型男女抗 B抗体效价与积分有显著性差异 ,女性明显高于男性。结论　提示在给肿瘤患者 ,特别是A型患者配血、鉴定血型时遇有正反不符、前后不符时要考虑是A抗原减弱 ,在给这些患者配血时 ,最好输注O型红细胞 ,已防输错血 ,防止发生意外。