Rh(D)阴性冰冻解冻红细胞的制备及临床应用

目的探讨Rh(D)阴性冰冻解冻红细胞的制备方法,并对临床输注效果进行分析。方法采用57%甘油作为冰冻保护剂,用9%NaCl、羟乙基淀粉40氯化钠注射液、0.9%NaCl洗涤制备冰冻解冻红细胞,按国家卫生部规定的检测方法对该制品进行质控,并对临床输注效果进行跟踪观察。结果Rh(D)阴性冰冻红细胞解冻后,各项质控指标符合国家标准,经临床输注后效果良好。结论该方法适用于临床上稀有血型患者的输血需要。     

云南省独龙族主要红细胞血型的分布

目的 了解云南省怒江州独龙族ABO和Rh等血型分布情况.方法 215名独龙族人群经试管直接凝集法分别检测了ABO、Rh、MN、P血型,采用血型群体遗传学研究方法,进行表现型及基因频率分析.结果 (1)ABO血型表现型频率顺序O>B>A>AB,基因频率r>p>q,民族指数=0.73.(2) Rh血型表现型以CCDee、C...     

RhD阴性冰冻红细胞临床应用

本血站自2000年7月开展了制备冰冻红细胞并应用于临床。5年来RhD(-)冰冻红细胞临床应用情况报告如下。表1RhD(-)冰冻红细胞使用情况三级医院二级医院合计n(%)n(%)n(%)A5826.04222.710024.5B7332.75027.012330.2O7533.67942.715437.7AB177.7147.6317.6RhD阴性冰冻红细胞临床应用@齐文革$保定市中心血站!河北保定071051 @刘冀华$保定市中心血站!河北保定071051 @杨珊珊$保定市中心血站!河北保定071051Rh血型;;RhD(-);;红细胞,冰冻…     

甘油对冰冻解冻红细胞质量的影响探讨

目的比较两种方法制备冰冻解冻去甘油红细胞质量差异。方法随机取2~6℃保存6d内的RH阴性去白细胞悬浮红细胞60袋,应用ACP215全自动血细胞处理仪制备甘油化红细胞,-80℃冰箱保存1~24个月,其中30袋去甘油冰冻保存,30袋不去甘油冰冻保存,然后取出于37~40℃水浴快速融化后,选择ACP215全自动血细胞处理仪去甘油程序进行去甘油洗涤处理,检测冰冻解冻去甘油红细胞的质量指标,并与国家标准进行比较。结果两种方法制备的冰冻解冻去甘油红细胞其血红蛋白水平、游离血红蛋白浓度、甘油残余量和细菌培养试验均满足《全血及成分血质量要求(GB18469-2012)》。结论两种方法均可制备符合国家标准的冰冻解冻去甘油红细胞。但从临床抢救角度看,采用冰冻前去甘油的方法,可以为临床抢救争取约30min的时间。     

关于冰冻红细胞三珠袋改进的建议

Rh（D）阴性血型在我国汉族人群中的比例为3‰～5‰，属于较稀有的血型，常以低温冰冻红细胞保存技术对其红细胞加以保存，多用甘油作为红细胞冰冻保护剂，但临床使用前必须解冻去除甘油，使红细胞内残留甘油含量〈1％才能在输注后获得适当的存活率，因此解冻后的洗涤过程中的操作至关重要。笔者在制备冰冻红细胞的过程中，对影响解冻冰冻红细胞质量的三珠袋做了一些改进，并通过实验对比证明了改进三珠袋的必要性和优点。     

Rh(D)阴性血在突发公共卫生事件中的应用探讨

目的 通过对模拟突发公共卫生事件中Rh(D)阴性血液成分制备及献血招募的应急演练,持续改进并加强对Rh(D)阴性血液临床供应的时效性及安全性。方法 以某医院1例产妇大出血,急需较大量Rh(D)阴性血液为模拟事件背景,根据血站《质量管理体系文件》《应急采供血预案》《Rh(D)阴性血应急采血及成分制备演练方案》《计算机信息管理系统应急预案》实施冻存的Rh(D)阴性冰冻加甘油红细胞制备,以血液制备时间,血液制备质量,Rh(D)阴性血献血员招募能力及异常情况下工作人员团队协作能力及应变能力为考核标准。结果 应急制备组人员在接到任务后能20min内到位,在规定4h内应急制备Rh(D)阴性冰冻解冻去甘油红细胞4U,血液制备过程均符合质量体系文件及标准操作规程要求,并能在模拟计算机管理系统瘫痪的情况下,按《计算机信息管理系统应急预案》的要求,将冰冻解冻去甘油红细胞手工打签、贴签、包装和放行等工作。采血科招募组在6h内组织16例在册登记的Rh(D)阴性血献血员至血站参加应急献血,经初步检测符合条件者10例,合计献血量3 300mL。结论通过建立Rh(D)阴性稀有血型红细胞冰冻库,创建符合条件的Rh(D)阴性献血员“流动血库”,成立应急预案,构建经培训及多次应急演练实践的Rh(D)阴性血液应急制备工作小组,通过加强血站成分制备科物料管理,仪器设备维护,能保障在突发公共卫生事件时及时为临床提供安全有效的Rh(D)阴性血液。     

ACP215处理机制备冰冻解冻去甘油红细胞与手工制备的对比研究

目的观察ACP215血细胞处理机洗涤制备冰冻解冻去甘油红细胞与手工洗涤制备的效果,并对其检测结果进行初步分析。方法采用48袋Rh（D）阴性全血,先甘油化冰冻保存,后解冻将其随机等分成2组,实验组采用ACP215血细胞处理机洗涤制备冰冻解冻去甘油红细胞,对照组采用手工开放式洗涤法进行制备。按照相关标准分别对实验组和对照组的红细胞回收率、游离血红蛋白含量、甘油含量、残留白细胞、体外溶血试验进行检测分析。结果实验组的红细胞回收率明显高于对照组,差异具有统计学意义（P〈0.05）,实验组的甘油含量、残留白细胞均低于对照组,差异具有统计学意义（P〈0.05）。但是两种方法制备产品中游离血红蛋白含量、体外溶血试验的差异无统计学意义（P〉0.05）。结论用ACP215血细胞处理机洗涤制备冰冻解冻去甘油红细胞操作简单易行,安全可靠,省时、省力,且产品质量均符合国家标准,有实际应用价值,值得推广。     

冰冻红细胞免疫功能的测定和分析

自从Siegal提出“红细胞免疫功能”概念以来,红细胞具有免疫功能(EIF)已为广大输血工作关注并逐渐深入了解。冰冻RBC输用前的质量标准常侧重于红细胞溶血率、回收率、输注体内存活率、上清甘油残余量,而对其ElF的变化及影响因素了解甚少。笔自1995年～2000年进行冰冻(-80℃)RBC的制备计143例,对其中部分洗涤RBC的成品进行了EIF的测定,现报告如下：     

冰冻保存Rh阴性稀有血液方法的建立

目的探索一种甘油化冰冻保存Rh阴性红细胞制备、融化及洗涤等环节的可行性方法。方法取22袋(1U/袋)Rh阴性全血,在制备甘油化红细胞时,采用电子摇摆秤和输血器自动控制法代替手工摇摆法控制甘油流速和流量;37℃～42℃水浴融化后,在洗涤Rh阴性冰冻红细胞时,采用9%氯化钠(NaCl)高渗溶液平衡后,再用0.9%NaCl等渗溶液洗涤二次,并计算红细胞的回收率及其相关指控指标。结果冰冻红细胞的回收率为(87.52±2.31)%,而甘油残余量、上清液游离血红蛋白(Hb)含量及体外溶血率分别为(6.76±2.38)g/L、(0.73±0.05)g/L及(9.78±1.85)%。结论采用上述简化方法制备、融化及洗涤冰冻红细胞,不仅缩短了时间,而且获得较满意的红细胞回收率,其他相关质控指标也均符合国家标准。     

血液储存时间对红细胞免疫功能的影响与临床安全输血的研究

目的探讨血液离体后在4℃保存过程中红细胞免疫功能的变化及对临床安全输血的影响。方法随机抽取50份常规CPD保养液充分混匀的全血,从细管小辫端取少量血样作为新鲜血样供当天检测,其余热合封闭成5段小样4℃保存备用(与血液相同条件保存);每次各取1个小样测定,分别于1、2、3、4和5周,对上述标本分别做红细胞C3b受体(RBC-C3b)和红细胞免疫复合物(RBC-IC)检测,同时在d0、d14和d28任取其中25份标本检测RBC超氧化物歧化酶(RBC-SOD)水平和酵母菌激活全血细胞免疫反应产生白细胞介素8(IL-8)的含量。结果血液保存时间在2周以内红细胞免疫功能有所下降,但不明显,随着保存时间的延长,RBC-C3b、RBC-SOD明显降低,d0、d14和d21 RBC-C3b RBC-SOD分别为18.84±2.23、16.54±2.42、14.28±2.15;为16222±4322、14111±4213、10459±389;而RBC-IC、IL-8则增高,在d0、d14和d28 RBC-IC分别为5.15±1.56、5.97±1.19、6.72±1.37;IL-8则为65.51±46.32、80.34±39.25、109.67±26.89。结论血液离体后在4℃保存过程中,随时间的延长,红细胞膜有所受损,表现为红细胞(部分)免疫功能降低。在大手术、老年人、严重的心脑血管疾病及因补体系统导致溶血性贫血等患者需要输血治疗时,尽可能输注较年轻(<14d)的血液有利于输血安全。     

Immunosuppressive effects of red blood cells on monocytes are related to both storage time and storage solution

BACKGROUND: Reduced monocyte function is associated with adverse outcomes from critical illness. Red blood cells (RBCs) are thought to impair monocyte function but relationships between RBC storage solution and monocyte suppression are unknown. This study was designed to test the hypothesis that immunosuppressive effects of RBCs on monocytes are related to both storage time and preservative solution. STUDY DESIGN AND METHODS: Monocytes from healthy adult donors were co‐cultured with RBCs that had been stored in AS‐1, AS‐3, or CPD only for 7, 14, or 21 days. Cells were then stimulated with lipopolysaccharide (LPS) and their supernatants assayed for tumor necrosis factor (TNF)‐α and interleukin (IL)‐10. Transwell experiments were performed to evaluate the role of cell‐to‐cell contact. Monocyte mRNA expression was quantified by real‐time–polymerase chain reaction. RESULTS: LPS‐induced TNF‐α production capacity was reduced compared to controls for all groups, but CPD‐only RBCs suppressed monocyte function more than RBCs stored in AS‐1 (p = 0.007) and AS‐3 (p = 0.006). IL‐10 production was preserved or augmented in all groups. A longer storage time was associated with reduced TNF‐α production capacity for AS‐1 and AS‐3 groups but not CPD. Preventing cell‐to‐cell contact did not eliminate the inhibitory effect of RBCs on monocyte responsiveness. RBC exposure was associated with decreased LPS‐induced TNFA mRNA expression (p < 0.05 for all groups). CONCLUSIONS: CPD‐only RBCs suppressed monocyte function more than RBCs stored with additive solutions. TNF‐α production was reduced even in the absence of cell‐to‐cell contact and was impaired at the mRNA level. Further work is needed to understand the role of preservative solutions in this process.     

悬浮红细胞储存时间对制备的洗涤红细胞钾离子和游离血红蛋白含量的影响

目的 了解不同储存时间的悬浮红细胞(悬红)制备的洗涤红细胞中钾离子浓度([K+])和游离血红蛋白含量(FHb).方法 将储存3 d(3 d组,n=6)、16 d(16 d组,n=6)及31 ～35 d(31 ～35 d组,n=7)的悬红,手工制备成洗涤红细胞,分别于制备前、制备后0、16和24h取样测定其[K+]和FHb.结果 3d组、16 d组和31 ～35d组悬红制备的洗涤红细胞24h时[K+]分别为(3.26 +0.57)、(3.39±1.12)及(2.97 +0.95) mmol/L(P ＞0.05),且均低于人体正常[K+]上限;3组制备的洗涤红细胞24 h时的FHb分别为(0.29±0.18)、(0.63±0.40)及(1.06±0.55)g/L,其中31 ～35 d组制备的洗涤红细胞的FHb明显高于3d组(P＜0.01).结论 悬红储存时间对制备的洗涤红细胞的[K+]影响不大,制备的洗涤红细胞24h保存期内的[K+]均低于人体正常参考值上限,但对FHb含量有较大影响.建议尽量避免使用接近储存期末的悬红制备洗涤红细胞.     

Hemolysis of red blood cells during processing and storage

BACKGROUND: During processing and storage, red blood cells (RBCs) undergo changes and cell injury resulting in hemolysis. Mostly, the separation of whole blood in top‐and‐bottom quadruple bag systems with break openings takes less than 4 minutes. However, longer separation times are not uncommon. The aims were to investigate whether hemolysis is increased in RBCs with longer separation time (RBCs_{>6 min}) compared to regular RBCs (RBCs_reg), to measure hemolysis increase during storage and to study whether frequency of hemolytic donations is donor dependent. STUDY DESIGN AND METHODS: RBCs_{>6 min} (n = 172) and 172 matched controls were tested for hemolysis on Days 1 and 21 RBC units from each group were stored at 4 ± 2°C and tested again after 5 weeks. Donor dependency was retrospectively investigated for 100 hemolytic RBC units. RESULTS: RBCs_{>6 min} exhibited a higher mean hemolysis rate than RBCs_reg (0.058% vs. 0.033%). Four RBC units were hemolytic (>0.8%), all RBCs_{>6 min} (2.36%). During storage, hemolysis in both groups increased with 0.24%. Hemolysis frequency did not seem to be donor dependent. CONCLUSIONS: Increased separation time is a useful screening tool for potentially increased hemolysis rate in RBCs. Hemolysis rate increased during storage equally in both groups. Hemolysis frequency appears donor independent.     

Chemokine receptors expressed on T cells in packed red blood cell units change over storage time

The transfusion of blood products is associated with adverse events that are related to the leukocytes in stored units of blood. These leukocytes have been shown to promote the elaboration of inflammatory cytokines. However, the status of a set of key inflammatory mediators, chemokine receptors, expressed on T lymphocytes in stored red blood cell (RBC) units is largely unknown. We investigated the expression pattern of selected chemokine receptors on T cells from non-leukocyte-reduced RBC units over storage time. Selecting segments from stored RBC units, we evaluated the T-cell subsets for the chemokine receptors CXCR3 and CCR4 by flow cytometry. Statistical analysis was performed by regression analysis. We analysed 30 samples stored between 5 and 38 days. The CD4+ T cells expressing CXCR3 increased by 0.27% daily (P= 0.02), whereas the expression of CCR4 declined by 0.40% daily (P < 0.001). Though the expression of the chemokine receptors on CD8+ cells followed the same trend, the changes were statistically nonsignificant. This study suggests that a longer duration of storage is associated with a higher expression of chemokine receptor CXCR3 and a lower expression of CCR4 on T cells in RBC units, suggesting a pro-inflammatory Th1 bias. The clinical significance of these changes in the setting of adverse transfusion events needs further evaluation.     

A method for freezing and washing red blood cells using a high glycerol concentration

A modification of previously reported methods for freezing red blood cells with a high concentration of glycerol is described. The limits of osmotic tolerance of human red blood cells range from one-half to four times isotonic. The introduction and removal of glycerol creates transient osmotic gradients that can exceed these limits. It is shown that much of the cell lysis following thawing and during washing can be related to hypertonic injury incurred during the glycerolizing procedure. Refinements of this portion of the process produce significant gains in recovery. Problems associated with washing out a high concentration of glycerol by centrifugal means can be alleviated by a preliminary dilution with hypertonic sodium chloride which serves also to reduce cell volume and increase cell specific gravity improving sedimentation characteristics and affording more latitude for the hypotonic stresses associated with glycerol removal. Careful attenion to the osmotic limits during both glycerolizing and washing results in a process in which hemolysis from freezing, thawing and washing can be reduced to less than three per cent.     

不同保存期全血制备洗涤红细胞的超微结构变化

目的 探讨不同保存期的全血制备洗涤红细胞前后电镜下红细胞形态的变化。方法 采集CPD—A抗凝全血。实验分Ⅰ～Ⅵ组，分别于4℃保存7、10、15、20、25、30d。于制备前取全血1滴，5％戊二醛固定，经2000r／min离心10min后，分出血浆，取样检测血红蛋白。余下的红细胞再加等量生理盐水，1500r／min离心5min，连续2次，取样为制备后测定组。结果 组Ⅰ和组Ⅱ制备前后红细胞成双面凹的圆盘结构，细胞均匀混悬。组Ⅲ制备前红细胞形态正常，制备后少数红细胞出现聚集状、球形或边缘不整齐，并有棘形红细胞出现。组Ⅳ、组Ⅴ、组Ⅵ于制备前血浆微红；制备后，球形红细胞、棘形红细胞、中问漏孔的红细胞增加。结论 制备洗涤红细胞的最佳时间应在4℃保存10d内的全血，保存15d以后的全血制备洗涤红细胞形态发生异常变化，出现棘形红细胞，囊泡化后的红细胞易溶血，红细胞寿命缩短，影响洗涤红细胞的质量。