过滤去除白细胞时温度对红细胞溶血的影响

目的:探讨在不同温度下过滤去除白细胞时对红细胞溶血的影响,寻找防止其溶血的措施。方法:将保存5d的红细胞悬液40 u随机分为对照组和试验组各20 u,前组红细胞悬液于4℃储血冰箱中取出后立即进行过滤,后组红细胞悬液置于35℃水浴箱复温10 m in再行过滤。测定各组红细胞悬液过滤前后的血浆K+和游离血红蛋白浓度。同时对保存5 d的650 u红细胞悬液(包括4℃立即过滤220 u红细胞悬液和经35℃复温10 m in后过滤430u红细胞悬液)滤后血浆的目测溶血情况进行回顾性分析。结果:对照组红细胞悬液滤后血浆K+、游离血红蛋白浓度分别为(6.41±0.16)mm o l/L、(37.84±6.15)m g/L,比滤前(5.62±0.15)mm o l/L、(23.31±4.24)m g/L明显升高(P<0.05);试验组滤后K+、游离血红蛋白浓度(5.72±0.16 mm o l/L、27.93±5.28 m g/L)与滤前(5.63±0.15 mm o l/L、23.51±4.26 m g/L)比较无显著性差异(P>0.05),但比对照组滤后明显降低(P<0.05)。两组滤前K+、游离血红蛋白浓度均无显著性差异(P>0.05)。4℃立即过滤与经35℃复温后过滤的红细胞悬液其溶血率存在显著性差异(P<0.05)。结论:现用的白细胞滤器能引起一定程度的红细胞溶血,红细胞悬液经35℃复温10m in后再行过滤可有效减少过滤引起的溶血。     

保存液添加氨基酸对试剂红细胞保存时间的影响

目的延长试剂红细胞在开放状态下的保存时间。方法在实验组红细胞保存液CPDA-Ⅰ中加入5%氨基酸,对照组仅为CPDA-Ⅰ保存液,并分别配制成5%的试剂红细胞,置4℃冰箱,连续观察18周:每周测定2组试剂红细胞的pH、Hb及K+的值。结果实验组在保存126 d后红细胞出现溶血,pH为(6.7±0.4)、Hb为0、K+为(1.16±0.04)mmol/L;而对照组仅保存28 d后红细胞出现溶血,pH为(6.6±0.2)、Hb为(4.8±1.0)g/L、K+为(1.24±0.12)mmol/L,2组只有Hb的差异有统计学意义(P<0.05)。结论在CPDA-Ⅰ中加入5%氨基酸可延长试剂红细胞的保存时间。     

SAGS红细胞悬液的实验研究

本文用 SAGS(氯化钠、腺嘌呤、葡萄糖、蔗糖)保存液及改良的 HogmanSAGM(氯化钠、腺嘌呤、葡萄糖、甘露醇)保存液分别重悬血比积(Hct)≥90％的浓缩红细胞,制成晶体盐红细胞悬液 SAGS-RCS 和 SAGM-RCS。系统地比较了两者在4±2℃贮存35天中的生物化学交化。结果表明,两者的体内存活率、ATP、2,3-DPG、pH及 K~+水平基本一致。但是,贮存35天时上清游离血红蛋白水平前者明显低于后者,分别为1.71±0.95g/L 和3.1±0.19g/L(P<0.01)。     

二甲基亚砜在人红细胞冻干前负载海藻糖过程中的作用

目的研究人红细胞冻干保存前负载海藻糖过程中二甲基亚砜(DMSO)的作用,优化红细胞负载缓冲液配方。方法实验组以浓缩红细胞25份(10ml/份)负载海藻糖,负载缓冲液中添加DMSO;对照组25份负载海藻糖,负载缓冲液中未添加DMSO。37℃条件下孵育8h后,分别检测两组红细胞胞内海藻糖负载量、胞外游离血红蛋白水平、ATP含量、红细胞变形性,并利用流式细胞术检测负载后红细胞膜的完整性。结果实验组与对照组红细胞的胞内海藻糖负载量分别为(57.033±4.883)mmol/L,(49.184±4.858)mmol/L(P<0.05);胞外游离血红蛋白浓度分别为(4.131±0.473)g/L,(5.410±0.501)g/L(P<0.05);ATP浓度分别为(3.874±0.426)μmol/g Hb,(3.358±0.306)μmol/g Hb(P<0.05);红细胞变形指数分别为0.330±0.0211,0.277±0.0232(P<0.01);红细胞胞膜PS表达率分别为(5.04±0.495)%,(8.69±0.862)%(P<0.01)。结论DMSO在红细胞负载海藻糖过程中可有效增加胞内海藻糖负载量,并显著改善负载缓冲液对红细胞胞膜的高渗损伤,更好地发挥海藻糖对红细胞的保护作用。     

深低温保存Rh阴性血液的质量控制

目的　确保深低温保存Rh阴性红细胞解冻后的质量。方法取ACD抗凝 ,4℃保存 6d内Rh阴性红细胞悬液或全血离心除去添加剂或血浆的浓缩红细胞 ,将红细胞经 (4 0 % )浓度甘油处理后 ,置 - 80℃冰冻保存。临床需要时 37℃水浴解冻复苏红细胞 ,依次用 9%NaCl羟乙基淀粉溶液 ,0 .9%NaCl羟乙基淀粉溶液 ,0 .9%NaCl各洗涤 1次。用生理盐水羟乙基淀粉溶液悬浮红细胞。结果 35袋冰冻解冻红细胞去甘油后红细胞回收率为(81 .1 7± 1 8.5 ) % ,游离血红蛋白为 (0 .6 3± 0 .1 6 ) g/L ,甘油残留量平均为 5 .3g/L ,体外溶血试验血红蛋白增加率为2 5 .1 9%。结论冰冻解冻去甘油红细胞各项指标均达到了国家规定的质量标准 ,临床输用效果良好     

原发性骨髓纤维化骨髓组织病理学:临床与血液学的关系及预后评价

作者对51例原发性骨髓纤维化(PMF)的骨髓活检作了观察,患者平均年龄64(17—86)岁,男34例。Hb10.5±3.3g/dl;白细胞14.9±14.1×10~9/L,Plt+260±219×10~9/L,血中白细胞前体细胞9.4±9.6％,幼稚红细胞3.0±4.7/100个白细胞,观察结果表明,不同病例有很大差异,甚至同一病例的不同     

贮存过期红细胞的生化功能及形态的恢复研究

目的研究如何使贮存过期红细胞的生化功能和形态恢复到正常水平。方法恢复前红细胞组为ACD保养液在(4±2)℃贮存3周过期3d洗涤2次的红细胞,将此红细胞分成等量的2组,1组加入由腺嘌呤、肌苷、丙酮酸钠、磷酸氢二钠混合组成的恢复液作为恢复组,另1组加入25ml0.9%NaCl作为盐水对照组,2组均37℃孵育1h,从上清液中离心分离获得囊泡;将采集献血者新鲜全血的同时分离的红细胞作为正常水平组。分别测定4组红细胞ATP、2,3DPG、MCV、形态学积分、渗透脆性程度及Hb浓度。结果ATP、2,3DPG及形态学积分:恢复组比恢复前组和盐水对照组均显著增加(P<0.001),与正常水平组无显著差异(P>0.05)。MCV:恢复组显著低于恢复前组及盐水对照组(P<0.001)。恢复组与盐水对照组红细胞在0.50%和0.55%NaCl处的低渗性溶血度比恢复前组显著减少(P<0.05),与正常水平组无显著差异(P>0.05)。恢复组囊泡内脱落的Hb比盐水对照组显著增多(P<0.05)。结论腺嘌呤、肌苷、丙酮酸钠、磷酸氢二钠组成的恢复液可使ACD(4±2)℃贮存过期3d的红细胞的生化功能和形态都恢复到正常水平;在恢复再生过程中,出胞的微小囊泡引起细胞膜部分脱落,导致红细胞的MCV变小。     

贮存式自体血输注在普外科择期手术中的临床应用研究

目的:研究贮存式自体血输注(preoperative autologous blood donation,PABD)在普外科择期手术中的临床应用效果。方法:筛选2017年11月-2018年8月本院普外科贮存式自体血输注70例,采用配对研究方法,将未采用贮存式自体血输注且术前基线资料无统计学差异的70例患者,配对作为对照组。比较2组患者异体红细胞和血浆输注量,围术期Hb和Plt变化,住院时间及住院费用的差异。结果:PABD组采血前、后Hb和Plt分别为138.26±14.73 vs 127.52±13.36g/L (P0.05)和(221.67±52.86 vs 198.35±52.65)×10~9/L (P0.05);2组患者围术期异体红细胞和血浆的输注量分别为0.20±0.71 vs 0.89±0.97 U和30.43±100.81 vs 106.52±152.61 ml(P0.05);2组患者术前Hb水平135.65±14.16 vs 134.15±11.98 g/L,术前Plt水平(270.36±58.28 vs 271.67±65.02)×10~9/L;术后1 d Hb水平120.24±14.40 vs 121.20±14.30 g/L,术后1 d Plt水平(241.80±63.58 vs 241.30±69.11)×10~9/L;术后3 d Hb水平123.15±13.80 vs 121.65±14.33 g/L,术后3 d Plt水平(251.26±72.94 vs 255.54±73.85)×10~9/L;出院前Hb水平122.78±13.92 vs 122.00±13.82 g/L,出院前Plt (262.50±80.96 vs 264.56±71.08)×10~9/L(P0.05);2组患者的住院时间14.84±3.37 vs 14.84±2.24(d)(P0.05),住院总费用和输血相关费用分别为50 627.27±9 889.45 vs 50 979.43±8 195.00元和354.39±362.57 vs 684.02±425.53元(P0.05)。结论:择期手术估计术中失血量1000ml的患者,贮存式自体血输注的应用可节约异体血输注,降低费用。     

Hb和Hct作为红细胞输注评估指标的回顾性分析

目的探讨Hb/Hct作为红细胞输注评估指标的作用与临床意义。方法收集本院2010年1月～2011年12月住院和门诊红细胞输注患者的输血信息及相关Hb检测结果,分析内科患者不同Hb区间红细胞输注量和Hb改变情况,以及手术科室不同输血类型和儿科患者红细胞输注量与输注前Hb值和改变情况。结果 27 913例次有效输血申请,输血前Hb平均值手术科室患者最高(85.23±20.23)g/L,儿科次之(83.60±21.13)g/L,内科最低(68.16±16.22)g/L;80%以上内科患者输注前Hb<80 g/L,且Hb越低,红细胞输注量随之略有增加;手术科室患者输注前Hb检测率仅为67.44%,且每例次输血申请中,红细胞输注量差异较多大,最高量为78 U,最低量为1U;非手术类输血Hb改变值明显高于手术类输血(含术中)(P<0.05)。结论 Hb/Hct可作为红细胞输注适应证评估及输注疗效评价的客观指标督导临床用血,但同时结合其他因素,如患者年龄、基础疾病、临床症状、失血量以及使用促造血细胞因子情况等,综合评判将更加全面、准确。     

重组人红细胞生成素治疗肿瘤化疗相关贫血的临床研究

目的:观察重组人红细胞生成素(rhEPO)治疗肿瘤化疗相关贫血的疗效。方法:根据NCCN《癌症及其治疗相关贫血治疗指南》,选择54例肿瘤化疗相关贫血患者(Hb<11g/L),所有病例均经病理或细胞学检查确诊。分为2组,治疗组32例,给予皮下注射rhEPO4000U/周,连用8周。对照组22例,不给予rhEPO治疗,仅给予益血生口服。分别于治疗2、4、6、8周后评定疗效。结果:治疗组和对照组基础Hb水平分别为(91±10)和(97±8)g/L;治疗后2周Hb增加分别为(10.2±5.3)和(3.8±5.9)g/L;治疗后4周Hb增加分别为(21.2±7.8)和(17.7±5.8)g/L;治疗后6周Hb增加分别为(26.2±6.4)和(23.7±6.8)g/L;治疗后8周Hb增加分别为(30.3±8.8)和(26.0±6.9)g/L。治疗组和对照组相比差异有统计学意义(P<0.01)。结论:rhEPO4000U/周对治疗肿瘤化疗相关贫血有肯定的疗效,能为顺利完成化疗提供有力的保障,而且安全患者能够耐受。     

再水化液因素对冰冻干燥保存后红细胞回收率的影响

目的　寻求一种能有效提高冰冻干燥 (简称冻干 )保存后人红细胞回收率的再水化体系。方法　测定1 0 %聚乙烯吡咯烷酮 (PVP)、6 %羟乙淀粉 (HES)、5 %羧甲基淀粉钠 (CMS)、生理盐水、0 75mol/L葡萄糖、等渗缓冲液及高渗缓冲液 (5×Buffer)的晶体渗透压和胶体渗透压 ;将浓缩红细胞和保护液混匀 ,预冻后移入冻干机内作冻干处理 ,冻干完毕后 ,用不同种类或不同温度的再水化液快速水化洗涤样本。结果　人红细胞冻干再水化后 ,6 %HES组、1 0 %PVP组和 5 %CMS组的红细胞回收率分别为 (93.6 5± 6 .1 8) %、(88.80± 9.4 9) %和 (91 .34± 8.1 3) % ,血红蛋白回收率分别为 (93.4 8± 4 .6 7) %、(89.0 2± 4 .6 7) %和 (88.79± 5 .35 ) % ,均极显著高于其他 4组[(1 5 .5 6± 1 2 .0 2 ) %～ (2 7.77± 6 .4 8) % ,(1 7.78± 1 0 .80 ) %～ (4 1 .5 0± 6 .4 3) % ) ](P <0 .0 1 ) ;不同温度的 6 %HES的再水化效果表明 ,再水化后 3个温度组的红细胞回收率无显著差异 ,但 37℃和 2 5℃组的血红蛋白回收率分别为(87.4 8± 5 .84 ) %和 (91 .37± 3.94 ) % ,均极显著高于 4℃组 (73.1 0± 5 .90 ) % (P <0 .0 1 )而且上清游离血红蛋白浓度也显著低于 4℃组。结论　再水化液的胶体渗透压对冻干保存后红细胞的保护作     

Red Cell Changes as Observed by a Simple Method for Continuously Recording Osmotic Fragility

Storage of human blood for six to eight weeks in ACD solution resulted in definite hemolysis which paralleled an increase in osmotic fragility determined by a new method. Mechanical injury as a result of delivering blood by airdrop markedly aggravated this hemolysis. Whole blood stored for six weeks in ACD solution with adenine added had less overt hemolysis, less osmotic fragility and better posttransfusion survival than blood from the same subject stored only in ACD solution. Comparison of changes in the osmotic fragility of stored red blood cells showed good correlation with in vivo survival measurements and suggested that this method of testing might be useful in programs to evaluate the effect of additives on the viability of banked blood.     

Red cell tolerance of admixture with heated saline

Red cell stability in the face of thermal stress has been evaluated only in the setting of prolonged incubation. This study was conducted to determine red cell tolerance of rapid mixture with heated saline, which exposes red cells to heat only until thermal equilibration, which is a matter of seconds. Half-units of 35-day-old red cells stored in CPDA-1 were mixed at 6 to 10 degrees C in the blood container with an equal weight of 60, 70, or 80 degrees C saline. This resulted in mean mixture temperatures of 30.9, 37.5, and 42.6 degrees C, respectively. Controls consisted of the same mixture, but with 6 to 10 degrees C saline. The red cells in the mixtures were assessed for osmotic fragility, and the supernatant was examined for plasma hemoglobin and potassium. Neither osmotic fragility curves nor supernatant hemoglobin or potassium changed significantly with saline temperatures of 60 or 70 degrees C. When 80 degrees C saline was used, osmotic fragility, supernatant hemoglobin, and potassium all increased significantly (p less than 0.01) over control values. Red cells tolerate rapid mixture with 70 degrees C saline without hemolysis or change in osmotic fragility.     

去白细胞悬浮红细胞储血袋两种放置方式的溶血性分析

目的 探讨去白细胞悬浮红细胞储血袋两种不同放置方式对上清液游离血红蛋白(FHb)和红细胞溶血率的影响,保证临床输血的安全、有效。方法 选择20例无偿献血者每人400 ml的全血,24 ml内制成去白细胞悬浮红细胞制品,将其等量均匀分装成10袋,随机分成2组,每组5袋,一组以直立放置,一组以水平放置,在同一环境条件下储存,分别在储存的第7天,14,21,28,35天时随机从各组取出1袋血液,分别测定其FHb和红细胞溶血率,进行统计学分析。结果 FHb和红细胞溶血率在贮存的第21天时检测的结果,直立放置组分别为(217.310±48.477)mg/L和(0.250±0.056)%,水平放置组分别为(173.972±39.027)mg/L和(0.189±0.045)%,直立放置组的结果要高于水平放置组,统计学结果分别为t=3.114,P=0.003＜0.05和t=3.798,P=0.001＜0.05,差异有统计学意义。结论 在血液储存期内,储血袋水平方式放置可减少红细胞破坏,对血液的贮存更加有利。     

Red blood cell endothelial nitric oxide synthase does not modulate red blood cell storage hemolysis

BACKGROUND: The red blood cell (RBC) endothelial nitric oxide synthase (eNOS) has been shown to regulate intrinsic RBC rheologic properties, such as membrane deformability, suggesting that a functional eNOS could be important in RBC viability and function during storage. This study examines the correlation between RBC eNOS deficiency and the propensity of RBCs to hemolyze under selected stress conditions including prolonged hypothermic storage. STUDY DESIGN AND METHODS: Fresh or stored RBCs from normal and eNOS knockout (KO) mice or from healthy human volunteers were subjected to selected hemolytic stress conditions including mechanical stress hemolysis, osmotic stress hemolysis, and oxidation stress hemolysis and evaluated during standard storage in CPDA‐1 solutions. RESULTS: Fresh RBCs from normal and eNOS KO mice demonstrated comparable susceptibility to hemolysis triggered by mechanical stress (mechanical fragility index 6.5 ± 0.5 in eNOS KO vs. 6.4 ± 0.4 for controls; n = 8‐9), osmotic stress, and oxidative stress. Additionally, RBCs from both mouse groups exhibited similar hemolytic profile at the end of 14‐day hypothermic storage, analogous to 42 days of human RBC storage. Storage of human RBCs (28 days in CPDA‐1) in the presence of NOS cofactors (l ‐arginine and tetrahydro‐l ‐biopterin) or inhibitor (N⁵‐[imino(methylamino)methyl]‐l ‐ornithine monoacetate) did not affect cell recovery or hemolytic response to the selected stressors. CONCLUSION: These studies suggest that RBC eNOS does not modulate susceptibility to hemolysis in response to selected stress conditions or prolonged hypothermic storage. Other strategies to increase nitric oxide (NO) bioactivity after prolonged storage utilizing NOS‐independent pathways such as the nitrate–nitrite–NO pathway may prove a more promising approach.     

Red blood cell osmotic fragility confidence intervals: a definition by application of a mathematical model.

The red blood cell osmotic fragility test is based on the measure of the resistance of red blood cells to lysis as a function of decreasing NaCl concentration. Up to now, several methods have been used for recording these data, but for the first time, the human red blood cell osmotic fragility confidence interval using the Orcutt mathematical model was determined. The absorbance of the hemoglobin measured at 540 nm, released by the red blood cells of 40 healthy adult individuals, was fitted to the equation Absorbance=p3 erfc ([NaCl] - p1/p2); p3 measures one half the absorbance produced by maximum red blood cell hemolysis, p1 is the [NaCl] producing 50% red blood cell hemolysis, and p2 is the dispersion in [NaCl] producing red blood cell hemolysis. Confidence intervals (mean+/-SD) for the three parameters were as follows: p1=4.2718+/-0.1848; p2=0.1947+/-0.0391, and p3=0.5568+0.0426. The usefulness of this osmotic fragility data analysis method using two pathological samples (beta-thalassemia minor and hereditary spherocytosis) was demonstrated. Parameters of the fitted data were compared with those obtained by the conventional recording method of Beutler.     

Prestorage leukoreduction and low-temperature filtration reduce hemolysis of stored red cell concentrates

BACKGROUND: Universal prestorage leukoreduction in Canada created the perception that stored red cells (RBCs) are more hemolyzed than their unfiltered predecessors. A pool-split design tested the effects of leukoreduction on hemolysis of stored RBCs. STUDY DESIGN AND METHODS: Two ABO-matched units were pooled, divided, and then processed into leukoreduced (LR) and nonleukoreduced (NLR) units with the Pall LT-WB or RC-PL systems and sampled during standard processing and storage for testing of sterility, counts, hemolysis, and osmotic fragility. RESULTS: Room temperature (RT) filtration of 10 pairs of LT-WB-LR and -NLR units showed significantly different percentage of hemolysis (0.39%) and osmotic fragility (0.643%) at 42 days. Cold-stored and -filtered units (2 days at 4 degrees C before processing) were less hemolyzed, but showed a similar proportional decrease of hemolysis in LR units (0.13% vs. 0.25% at 42 days). RBCs from RC-PL systems showed the lowest hemolysis although there was a filtration effect (0.05% vs. 0.12%, 42 days). Osmotic fragility paralleled hemolysis. Segment samples gave inaccurate results. Two-day prefiltration cold storage reduced hemolysis from 0.36 to 0.07 percent (42 days, p < 0.001). RT-LR hemolysis became significantly higher by Day 10 and 4 degrees C LR by Day 12. NLR units showed hemolysis by Day 7. LR units filtered cold were less hemolyzed (p < 0.05) than RT-LR but osmotic fragility was unchanged. CONCLUSIONS: LR-RBCs prepared by any of three methods (LT-WB, RT or cold; RC-PL), filtered at 4 degrees C, were less hemolyzed during storage than nonfiltered concentrates: 4 degrees C leukoreduction is beneficial for RBCs and does not cause hemolysis or enhance fragility.     

海藻糖负载红细胞及其冻干保存研究

为了研究海藻糖负载红细胞方法的可行性及红细胞内海藻糖对冻干红细胞的影响,利用红细胞膜在37℃时细胞膜上部分脂质由固态变为液态、流动性增大和膜通透性增加的性质,将红细胞置于高浓度海藻糖负载液中孵育7小时,并以磷酸缓冲盐溶液中孵育的红细胞作为对照,对红细胞的海藻糖负载率、形态学、渗透脆性、变形性、ATP含量及2,3-DPG含量进行评价。结果表明:负载后红细胞内海藻糖含量为36.56±7.95mmol/L,实验组红细胞溶血率为(15.663±3.848)%,对照组红细胞溶血率为(5.03±1.85)%,差异显著(P<0.05);实验组红细胞变形指数是0.0289±0.00738,对照组红细胞变形指数是0.1200±0.0121,差异显著(P<0.05);负载后实验组红细胞内ATP含量为2.67±0.54μmol/gHb,对照组红细胞内ATP含量为5.22±1.10μmol/gHb(P>0.05),实验组红细胞渗透脆性降低,明显低于对照组。尽管负载组的红细胞大小不一,形态各异,但在透射电镜下绝大多数红细胞膜完整,胞内血红蛋白密度均匀,而对照组有近一半的细胞膜不完整并有漏孔,胞内血红蛋白密度变浅。实验组与对照组中2,3-DPG含量均为零。实验组红细胞冻干再水化后,血红蛋白回收率46.44±4.14%,对照组血红蛋白回收率8.33±2.34%,差异显著(P<0.001)。结论:海藻糖负载的红细胞功能符合输注标准,负载方法可行,负载入细胞内的海藻糖能够保持细胞膜的完整性,大大提高了冻干红细胞的回收率,为红细胞的冷冻干燥成功迈出了第一步。