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     

冰冻保存红细胞简化洗涤程序的研究

目的　研究冰冻保存后红细胞沉降去甘油的方法。方法　 6份全血采自健康献血者 ,ACD抗凝 ,4℃保存 1d ,离心获压积红细胞 ,加入甘油到终浓度为 4 0 % ,- 80℃保存 7d。 4 2℃条件下冻融红细胞 ;分两组进行洗涤去甘油 ,离心组 :常规法洗涤红细胞 4遍。沉降组加入 9%NaCl、6 %羟乙基淀粉和 5 %羧甲基淀粉钠 ,沉降 30min ,弃上清 ;加入 6 %羟乙基淀粉、0 .9%NaC1和 6 %CMS ,沉降 2 0min ,去除上清 ;加入 0 .9%NaC1和 5 %羧甲基淀粉钠 ,沉降 15min ,去除上清 ;重复步骤 ;以生理盐水重悬红细胞。结果　洗涤后红细胞形态正常 ,离心洗涤和沉降洗涤后红细胞的回收率 (% )分别为 87.95± 8.6 0和 89.2 5± 4 .17、上清液中游离血红蛋白的含量 (g/L)为 0 .15± 0 .0 9和0 .2 3± 0 .19,上清液中晶体渗透压 (mOsm)为 2 90 .83± 4 .17和 2 95 .6 7± 8.0 7、ATP含量 (μm/ gHb) 34. 0 2± 9.0 7和 34.70± 11.0 1,红细胞的变形指数分别为 0 .5 9± 0 .0 9和 0 .6 1± 0 .0 6 ,统计学处理上述各项指标 ,两组实验比较 ,差异无显著性。结论　冰冻保存后的红细胞 ,以沉降法去甘油与常规离心法效果无显著差别 ,细胞质量符合血库的质量要求 ,同时具有操作简便的优点。     

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     

新型冰冻红细胞洗涤机洗涤效果的研究

目的研制一种全自动、封闭式,体积小、重量轻,洗涤程序设计简单的新型冰冻红细胞洗涤机。方法全血采自健康献血者,ACD抗凝,离心获浓缩红细胞,加入甘油到终浓度为40%,-80℃保存。42℃条件下冻融红细胞,利用自行设计的透析式新型冰冻红细胞洗涤机,使用3种自动化洗涤程序洗涤红细胞,直至可输注。结果洗涤后红细胞形态正常,3种程序洗涤后红细胞的回收率(%)分别为69.73±8.36,85.09±6.03和88.43±4.72;红细胞血红蛋白的回收率(%)分别为45.30±2.66,70.80±3.76和81.26±5.55;洗后上清液中游离血红蛋白的含量(g/L)为0.20±0.08,0.14±0.03和0.25±0.13,采用单因素k水平方法对数据进行统计学分析,得出了最佳洗涤程序。结论该新型冰冻红细胞洗涤机采用优化的洗涤程序,洗涤后的冰冻红细胞达到了国家的质量标准。     

海藻糖和蔗糖在人红细胞冰冻干燥保存中的效果比较

目的 比较海藻糖和蔗糖在人红细胞冰冻干燥保存过程中对细胞的保护效果。方法 将浓缩红细胞、含有不同浓度海藻糖或蔗糖的30％聚乙烯吡咯烷酮(PVP)按照1:3(V/V)的比例混合,-80℃冰箱中预冻1h,入冻干机内冻千处理后,用37℃等渗缓冲液快速水化洗涤样品,然后将其分为3个处理组:组1为30％PVP组(简称PVP组),组2、3为含有5％、10％和15％海藻糖或蔗糖的30％PVP组(简称海藻糖组或蔗糖组),测定各项指标。结果 冻干红细胞再水化后,海藻糖组的细胞回收率[(18.86±4.63)％、(21.73±7.32)％和(18.01±4.53)％]显著低于PVP组[(45.97±11.77)％](P<0.01);蔗糖组,蔗糖浓度为5％、10％时,细胞回收率分别为(37.40±2.90)％和(37.77±4.78)％,二者显著低于PVP组(P<0.05),但当蔗糖浓度升至15％时,细胞回收率[(42.54±10.25)％]和PVP组无显著差异;血红蛋白回收率,海藻糖组[(50.00±3.47)％、(40.91±9.09)％和(52.09±7.01)％]显著低于PVP组[(77.27±3.63)％](P<0.05),而蔗糖组和PVP组无显著差异,且显著高于海藻糖组(P<0.05)。洗涤后3组的游离血红蛋白浓度均<1g/L,其余各项指标的对比关系类似于水化后指标。结论 在冻干保护液中添加不同浓度的海藻糖或蔗糖对于提高冰冻干燥保存后红细胞的回收率和血红蛋白浓度作用不明显;但     

联合输注冰冻单采血小板与新鲜冰冻血浆治疗急性大出血的比较

严重创伤急性大出血患者失血量大、病情危重,输血抢救是主要方法之一。但短期内输入大量晶体盐、人造胶体及库存红细胞,会引起稀释性血小板和凝血因子减少,导致凝血功能障碍而难于止血。因新鲜血小板在(22±2)℃轻振荡保存有效期短,采集检验费时,不能满足临床危重患者急救需要。我们采用冰冻单采血小板与新鲜冰冻血浆(FFP)联合输注(部分病例加冷沉淀),报告如下。1资料与方法1.1临床资料:35例患者输注悬浮红细胞>10U时,随机分为输注冰冻单采血小板组(冰冻单用组,17例)与冰冻单采血小板联合FFP输注组(冰冻联合组,18例),其中男21例,女14例;…     

亚甲兰光化学法红细胞病毒灭活的初步探讨

目的　探讨择定条件下亚甲兰光化学方法对红细胞悬液的病毒灭活效果及其对红细胞质量指标的影响。　方法　在国产 PVC储血袋内的 GMA红细胞悬液中预置 VSV和 Sindbis病毒 ,加入终浓度为 5μmol/L的亚甲兰后用380 0 0 lx荧光照射 60分钟 ,测定处理后红细胞悬液及其匀浆中的病毒滴度及保存 2 1天内各时间段的红细胞 ATP、2 ,3-DPG、上清 K+ 和游离血红蛋白等质量指标。　结果　经上述方法处理后 ,红细胞悬液上清和红细胞匀浆中的 VSV病毒滴度分别降低 8.87± 0 .4 2 lg TCID50 和 7.79± 0 .90 lg TCID50 ;Sindbis病毒滴度则分别降低 6.5 8± 1 .67lg TCID50 和 5 .0 8±1 .4 2 lg TCID50 :处理后保存 2 1天内的各时间段红细胞主要质量指标未发生明显不良变化。　结论　亚甲兰光化学方法能有效灭活红细胞悬液及红细胞匀浆中与细胞相关的脂质包膜模型病毒 ,对红细胞体外质量指标无明显不良影响     

2组洗涤溶液制备冰冻解冻去甘油红细胞的效果比较

目的观察2组洗涤溶液制备冰冻解冻去甘油红细胞的结果,比较洗涤溶液组合中是否加入羟乙基淀粉20氯化钠溶液对洗涤结果的影响。方法取44份全血,应用ACP215血液处理仪制备成冰冻红细胞,分A组:9%NaCl溶液、羟乙基淀粉20氯化钠注射液、0.9%NaCl溶液;B组:9%NaCl溶液、0.9%NaCl溶液2种洗涤溶液进行解冻去甘油。对2组红细胞回收率、残留白细胞、残留血小板、甘油含量、游离血红蛋白含量、体外溶血等指标进行检测分析。结果 A组解冻红细胞的红细胞回收率(86.0±3.4)%、残留白细胞(0.57±0.18)%、残留血小板0、甘油含量(3.4±0.8)g/L、游离血红蛋白含量(0.55±0.14)g/L、体外溶血试验(12±5)%均符合国家标准,B组红细胞回收率(87.0±2.3)%、残留白细胞(0.51±0.24)%、残留血小板0、甘油含量(3.5±0.7)g/L、游离血红蛋白含量(0.47±0.10)g/L、体外溶血试验(10±4)%也符合国家标准,2组解冻红细胞制品也均符合AABB标准和欧洲标准中对于红细胞回收率的要求,2组结果差异无统计学意义,P>0.05。结论羟乙基淀粉20氯化钠注射液对使用ACP215血液处理仪制备冰冻解冻去甘油红细胞的洗涤效果无影响,但洗涤液中不加入羟乙基淀粉20氯化钠溶液,其制品质量符合国家标准、AABB标准和欧洲标准,更加经济、省时。     

25～45Gyγ射线辐照对红细胞制品质量的影响

目的　研究不同剂量辐照对红细胞的活性及功能的即刻损伤及保存损伤 ,确定辐照血液的保存时间。方法　将CPDA 全血 4 0 0ml分成 4组 ,于采血后 1d进行 0 (为对照组 )、2 5、35、4 5Gyγ射线辐照。在辐照后 0、4、7、1 4、2 1、2 8、35d取样测定红细胞活性、功能指标。结果　 0～ 35d保存过程中 ,①红细胞功能 :各辐照组的 2 ,3 DPG含量与对照组比较均无显著性差异 (P >0 .0 5 ) ;②红细胞活性 (ATP含量 ) :2 5Gy辐照组在 35d保存过程中与对照组差异无显著性 (P >0 .0 5 ) ;35、4 5Gy组分别于 35d(84 .7% )及 2 8d(83.6 % )起低于对照组 (P <0 .0 5 ) ;③红细胞脆性 :红细胞最小抵抗力 ,2 5、35、4 5Gy组分别自 35、2 8、2 1d起低于对照组 (P <0 .0 5 ) ;红细胞最大抵抗力 ,保存过程中 2 5Gy组与对照组差异无显著性 (P >0 0 5 ,35、4 5Gy组分别自 35及 1 4d起低于对照组 (P <0 .0 5 ) ;④游离K+ :2 5Gy组自辐照后 1d ,35、4 5Gy组自辐照后 0d起即高于对照组 (P <0 .0 5 ) ;K+ 含量与剂量呈正相关性 (r为 0 .96 6～ 0 .999) ;⑤游离血红蛋白 :在保存过程中显示出一定的剂量关系 ,但差异尚无统计学意义。结论　辐照对红细胞有一定的损伤 ,损伤程度与剂量有一定的相关性 ,但总体来说损伤不大 ,对红细胞活性     

去白细胞悬浮红细胞与悬浮红细胞储存期内溶血率的比较

目的观察去白细胞悬浮红细胞和常规制备悬浮红细胞在储存期内的溶血率变化,评价储存前白细胞过滤对于红细胞溶血率的的影响。方法随机采集80袋200 mL全血,将其分为白细胞过滤组和未过滤组,其中过滤组分别用上海输血技术公司、威高和南格尔3家公司的血袋各采20袋,过滤并制成悬浮红细胞;未过滤组用上海输血技术公司普通血袋采20袋,制成悬浮红细胞。2组置于(4±2)℃条件下储存35 d,在储存期14、21、28、35 d取样检测红细胞溶血率。结果 60袋去白细胞悬浮红细胞储存期内平均红细胞溶血率分别为14 d(0.11±0.06)%、21 d(0.20±0.10)%、28 d(0.31±0.13)%、35 d(0.42±0.15)%,20袋未过滤组平均红细胞溶血率分别为14 d(0.07±0.08)%、21 d(0.10±0.06)%、28 d(0.15±0.09)%、35 d(0.22±0.11)%,过滤保存21 d后溶血率有显著升高(P<0.05),但2组结果均符合欧盟和AABB标准中对于储存期末红细胞溶血率的要求。3家公司血袋制备的去白细胞悬浮红细胞保存期末溶血率差异无统计学意义(P>0.05)。结论储存前白细胞滤除过程未对35 d储存期的红细胞溶血率产生明显影响,35 d储存期的红细胞符合欧盟标准和AABB标准。     

The extension of 4 degrees C storage time of frozen-thawed red cells

Blood was drawn from volunteer donors and frozen using the high glycerin, mechanical freezing procedure accepted by the United States Navy. Subsequently, the units of blood were thawed and washed. Various anticoagulants were added, and the red cells were stored in a refrigerator at 4 degrees C for periods of up to 28 days. Chemical analyses were performed periodically. These showed that the addition of the anticoagulants ACD, CPD and CPDA-1 caused the red cells to be preserved better than the currently accepted 0.9-percent NaCl, 0.2-percent glucose solution. In vivo 51Cr viability studies performed on blood stored with CPDA-1 for 14 days showed a 24-hour viability of 78.8 +/- 8.4 percent. In a subsequent study, the blood was stored for 21 days prior to freezing and then was rejuvenated and frozen. The cells were thawed, washed, and stored at 4 degrees C with CPDA-1 for an additional 14 days. The 24-hour viability of these cells was determined to be 74.0 +/- 5.1 percent. These findings show that the postthaw storage time of red cells can be increased greatly over the now-accepted 24 hours, if bacterial sterility can be assured.     

Phosphate ion exchange resin used in the liquid preservation of baboon red cells

Baboon whole blood, collected in 14 percent citrate-phosphate-dextrose anticoagulant solution in plastic bags was stored in 100-ml aliquots at 4 degrees C for 28 days in the presence or absence of 0.75 grams of phosphate anion exchange resin. In vitro and in vivo measurements after autologous transfusions were made to determine whether the phosphate anion exchange resin had any beneficial effect on the blood during storage. The in vitro measurements of red cell 2,3-diphosphoglycerate and P50 were higher throughout the 28 days of storage at 4 degrees C in the blood stored in the phosphate anion exchange resin. After autologous transfusions in six baboons of red cells prepared from whole blood stored at 4 degrees C for 21 days, the 24-hour posttransfusion survival values were 86 +/- 6 percent (mean +/- SD) in the presence of resin and 83 +/- 6 percent in the absence of resin. In five other baboons, red cells prepared from 28-day-old blood showed a mean 24-hour posttransfusion survival of 82 +/- 4 percent in the presence of resin and 75 +/- 4 percent in the absence of resin. The addition of a phosphate anion exchange resin to the citrate-phosphate-dextrose anticoagulant provided better maintenance of red cell 2,3-diphosphoglycerate concentrations and P50 levels during storage of whole blood at 4 degrees C, and red cells prepared from whole blood stored in this solution had better oxygen transport function than red cells prepared from blood stored without resin. Red cell adenosine triphosphate concentrations and 24-hour posttransfusion survival values were similar whether or not the anticoagulant contained resin.     

Survival and biochemical characteristics of stored red cells preserved with citrate-phosphate-dextrose-adenine-one and two and prepared from whole blood maintained at 20 to 24 degrees C for eight hours following phlebotomy

Studies were conducted to evaluate the characteristics of red cells stored for 35 days following preparation from units of whole blood anticoagulated with citrate-phosphate-dextrose-adenine-one and two (CPDA-1 and CPDA-2) and maintained at 20 to 24 degrees C for 8 hours after phlebotomy. The mean (+/− 1 SD) 24-hour survival for transfused CPDA-1 autologous red cells with hematocrit levels of 78.1 +/− 2.3 percent was 78.0 +/− 8.1 percent (n = 9). The 24-hour survival of red cells from units preserved with CPDA-2 with hematocrit levels of 79.3 +/− 4.5 percent was 74.8 +/− 8.6 percent (n = 15). This difference in survival was not significant. Red cells from control units stored for 1 day showed a 24-hour survival of 91.9 +/− 4.2 percent (n = 7). During the 8-hour holding, red cell adenosine triphosphate levels increased by 15 to 25 percent in units drawn into both CPDA-1 and CPDA-2. After the initial 8-hour period, the red cell 2,3 diphosphoglycerate levels were 54 +/− 12 percent (mean +/− 1 SD) of initial levels in units drawn into CPDA-1 and 58 +/− 8 percent of initial levels in units drawn into CPDA- 2. Following 35 days of storage, units of red cells prepared from whole blood drawn into CPDA-1 and CPDA-2 had comparable plasma cation and ammonia levels and similar amounts of cell-free hemoglobin. These data indicate that red cells can be stored satisfactorily for 35 days when prepared from whole blood held at 20 to 24 degrees C for 8 hours.     

人红细胞冻干前负载海藻糖最佳化研究

为更好的实现海藻糖在红细胞冻干保存中的保护作用,关键是克服质膜对海藻糖的非渗透性,使胞质内海藻糖达到有效浓度。本研究的目的是通过对人红细胞负载海藻糖的规律性研究,筛选出海藻糖负载的最佳负载条件并评价海藻糖负载对红细胞各项理化指标的影响。在不同孵育温度(4、22和37℃)、孵育时间(0、2、4、6、8、10小时)、不同负载缓冲液浓度(0、200、400、600、800、1000mmol/L)条件下检测新鲜红细胞对海藻糖的成功负载量及红细胞各项理化指标;在固定负载条件下,对新鲜红细胞和4℃保存72小时红细胞海藻糖负载、游离血红蛋白(FHb)、血红蛋白(Hb)和红细胞平均体积(MCV)进行了比较。结果表明:红细胞对海藻糖的负载与孵育温度、时间及负载缓冲液海藻糖浓度密切相关。随着温度的升高、时间的延长和负载缓冲液海藻糖浓度的增加,红细胞对海藻糖的摄取量也随之增加。在海藻糖负载最佳条件下,新鲜红细胞和4℃保存72小时红细胞的胞内海藻糖浓度、FHb浓度分别为65.505±6.314mmol/L、66.2±5.002mmol/L和6.567±2.568g/L、16.168±3.922g/L。结论:红细胞负载海藻糖的最佳条件是采用新鲜红细胞,在37℃条件下、海藻糖浓度为800mmol/L的负载缓冲液中孵育8小时,这一条件可使胞内海藻糖达到有效浓度,并保持红细胞细胞理化性质稳定和膜完整性。     

The effect of prestorage irradiation on posttransfusion red cell survival

Transfusion-associated graft-versus-host disease (TA-GVHD) may occur whenever immunologically competent allogeneic lymphocytes are transfused to an immunocompromised recipient. Irradiation of blood components eliminates the risk of TA-GVHD but may damage the cellular elements in the transfused component, particularly if the cells are stored for prolonged periods in the irradiated state. To study the effect of irradiation on long-term storage of red cells, AS-1 red cells from eight normal subjects were prepared on two occasions. On one occasion, the units were stored as standard AS-1 red cells for 42 days at 4 degrees C; on the other, they were exposed to 3000 cGy radiation within 4 hours of collection and then were stored as AS-1 red cells for 42 days at 4 degrees C. The donations were at least 12 weeks apart. Irradiated units demonstrated significant elevations in poststorage plasma hemoglobin (Hb) (623 +/- 206 vs. 429 +/- 194 g/dL [6230 +/- 2060 vs. 4290 +/- 1940 g/L], p less than 0.02) and plasma potassium (78 +/- 4 vs. 43 +/- 9 mEq/L [78 +/- 4 vs. 43 +/- 9 mmol/L], p less than 0.01) and significant decreases in red cell ATP (1.9 +/- 0.2 vs. 2.1 +/- 0.3 microM/g Hb, p less than 0.04) and 24-hour posttransfusion red cell recovery (68.5 vs. 78.4%, p less than 0.02), as compared to nonirradiated units. It can be concluded that irradiation with 3000 cGy damages red cells and that long-term storage in the irradiated state may enhance this damage. Red cells should not be stored for 42 days after irradiation with 3000 cGy.     

Refrigerated storage of lyophilized and rehydrated, lyophilized human red cells

Human red cells (RBCs) were collected in CPDA-1 and then freeze-dried in lyoprotective solution. The lyophilized RBCs were then stored at -20 degrees C for 7 days. At the end of the storage period, the lyophilized RBCs were rehydrated and washed in dextrose saline. The washed, reconstituted, lyophilized RBCs were resuspended in final wash solutions of ADSOL, CPDA-1, or a special additive solution containing glucose, citrate, phosphate, adenine, and mannitol, and then they were stored at 4 degrees C for an additional 7 days. The main purpose of this study was to determine whether human RBCs can be lyophilized in such a manner that normal metabolic, rheologic, and cellular properties are maintained during rehydration and subsequent storage in standard blood bank preservative solutions. Our results show that reconstituted, lyophilized RBCs maintained levels of ATP, 2,3 DPG, lactate, and cellular properties that are equal to or better than those in control nonlyophilized RBCs stored for a comparable period in CPDA-1. Reconstituted, lyophilized RBCs stored at 4 degrees C after rehydration also show better maintenance of ATP, 2,3 DPG, and lactate than do control RBCs stored in the same preservative solutions for comparable periods.     

预冻温度和冻干机搁板温度对冷冻干燥红细胞的影响

为研究预冻温度和冷冻干燥机 (冻干机 )搁板温度对红细胞冻干保存后回收率的影响 ,采用主要成分为7%二甲亚砜和 4 0 %聚乙烯吡咯烷酮 (PVP)的缓冲液作为保护液 ,在不同预冻温度或搁板温度下进行红细胞的冻干保存。首先将新鲜血液离心、洗涤和平衡以制备浓集红细胞 ,然后将浓集红细胞和保护液按 1∶3混匀制备红细胞悬液 ,在不同温度 (- 2 0 ,- 35 ,- 4 5 ,- 80或 - 196℃ )下预冻后移入冻干机 (搁板温度设为 - 35℃ ,抽真空压力为2 0 0mbar)内进行真空干燥。为研究冻干机搁板温度对冻干红细胞的影响 ,将上述红细胞悬液先在 - 80℃下预冻后移入冻干机 ,在不同的搁板温度 (- 2 0 ,- 2 5 ,- 30 ,- 35 ,- 4 0或 - 4 5℃ )下抽真空干燥 ,冻干结束后用 37℃的再水化液快速水化。结果表明 :在不同预冻温度下 ,冻干后红细胞和血红蛋白的回收率均在 85 %和 75 %以上 ,各组之间差异不显著。但 - 196℃组上清游离血红蛋白浓度显著高于其他各组 (P <0 .0 1) ;当搁板温度等于或高于- 2 5℃时 ,样品无法冻干 ;当搁板温度等于或低于 - 30℃时 ,随着搁板温度的降低 ,冻干时间相应延长。再水化后红细胞和血红蛋白回收率均在 90 %以上 ,各组之间差异不显著 ;但当洗涤至等渗时 ,4组血红蛋白回收率之间差异不显著。结论 :本冻     

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。     

不同冻干保护体系对海藻糖负载红细胞冻干保存影响的研究

本研究旨在评价冻干保护剂人血白蛋白、葡聚糖、聚乙烯吡咯烷酮和甘油对海藻糖负载后红细胞冰冻干燥保存的影响,筛选最佳冻干保护体系。将浓缩红细胞在37℃,浓度为800 mmol/L的海藻糖溶液中孵育7 h,经PBS液冲洗3遍后制成海藻糖负载的浓缩红细胞。对照组为海藻糖负载红细胞不添加保护剂,直接冻干;实验组将人血白蛋白、葡聚糖、聚乙烯吡咯烷酮、甘油等组成的冻干保护体系与海藻糖负载浓缩红细胞混合,两组样品在常温下平衡30 min,移入-80℃深低温冰箱,预冻24 h,入冻干机冻干处理24 h。用温度为37℃,6%羟乙基淀粉40注射液快速再水化样品,用氰化血红蛋白试剂盒测定血红蛋白溶血率,计算血红蛋白回收率,同时测定干燥样品含水量。结果表明：当样品含水量在3%-4%时,对照组冻干红细胞血红蛋白回收率为（33.57±2.89）%,白蛋白组血红蛋白回收率为（51.15±1.98）%,差异有显著性意义（P〈0.05）。选用不同浓度的葡聚糖为冻干保护剂,血红蛋白回收率较对照组明显降低,随浓度增加,血红蛋白回收率逐渐升高,当浓度为36%时,血红蛋白回收率为（22.15±4.12）%,差异有显著性意义（P〈0.05）。不同浓度的聚乙烯吡咯烷酮（PVP）组成的冻干保护体系,当浓度小于40%时,血红蛋白回收率明显低于对照组,差异有显著性意义（P〈0.05）。10%甘油组血红蛋白回收率为（3.93±1.80）%,差异有显著性意义（P〈0.05）。结论：人血白蛋白在海藻糖负载的冻干红细胞中发挥重要保护作用,葡聚糖与浓度小于40%PVP可削弱细胞内海藻糖的保护作用。液态的甘油不宜作为红细胞冰冻干燥保存的保护剂。     

用α-半乳糖苷酶制备可供输注的人通用型O型红细胞

研究的目的是将人群中28％的B型血改造成通用O型血,提高O型血的储存和使用比例,以缓解战争、恐怖袭击、突发事件等紧急状态下对O型血的大量需求用α-1.3半乳糖苷酶作为B→O血型改造的工具酶,摸索改造一个使用单位B型血红细胞(100m1)的最佳条件,结果实现了在密闭、无菌条件下使用50U／ml工具酶．pH5．6,26℃ 2小时,进行B→O的血型改造。结论：改造后的通用型O型血红细胞符合生物制品检定规程的要求．并可在4℃存放21天