Evaluation of the red cell storage lesion after irradiation in filtered packed red cell units

Packed red cell units (n = 10) were filtered and divided equally. One-half unit from each donor was irradiated (x) (3500 cGy). On Days 0, 14, 28, and 42, ATP, K+, Na+, lactate dehydrogenase (LDH), plasma-free hemoglobin (PFH), and pH were determined. The reduction in ATP was greater in the irradiated than the nonirradiated (y) units by Day 42 (mean x-y: -70, p = 0.0005). The increase in K+ was greater in the irradiated than nonirradiated units on Days 14, 28, and 42 (mean x-y: 17-20, p = 0.0001). Decrease in pH and increases in LDH and PFH were significant (p less than 0.05) on Day 42 only. K+ increases added only 1.7 to 2.0 mmol per unit, a difference felt to be clinically insignificant. The changes noted in ATP, pH, LDH, and PFH are significant but minimal on Day 42 and imply that viability changes would also be minimal. These biochemical data support the storage of irradiated units for at least 28 days.     

Effect of 24-hour storage at 25°C on the in vitro storage characteristics of CPDA- 1 packed red cells

BACKGROUND: Packed red cells (RBCs) warmed above 10 degrees C are generally discarded. Few data exist on the degree of accelerated metabolism and increased hemolysis of packed RBCs allowed to warm. STUDY DESIGN AND METHODS: Twenty-four CPDA-1 packed RBC units were combined in 3-unit pools and subdivided into 2 test units and a control unit. One test unit from each pool was warmed to 25 degrees C for 24 hours on Day 6 and the other test unit was warmed on Day 20; control units were maintained at 1 to 6 degrees C. RBC and supernatant chemistries and RBC morphology were measured weekly (Days 0, 7, 14, 21, and 28) and on the day before warming (Days 6 and 20). RESULTS: Warming CPDA-1 packed RBCs accelerated the catabolism of glucose 10-fold and produced concentrations of glucose, lactate, and ATP after 25 days of storage that were equivalent to those in unwarmed units at 35 days. Supernatant sodium and potassium concentrations were corrected partially with warming. RBC morphology transiently normalized with warming and without increased hemolysis; no bacteria growth was detected. CONCLUSION: One day of 25 degrees C storage of CPDA-1 packed RBCs accelerates essential metabolite break-down equivalent to 10 days of storage at 1 to 6 degrees C. It does not appear to matter whether the packed RBCs are warmed on Day 6 or Day 20. This information may be useful in determining the acceptability of blood allowed to warm above 10 ° C.     

Viability and in vitro properties of AS-1 red cells after gamma irradiation

BACKGROUND: Irradiation has been shown to adversely affect both in vivo 24-hour recovery (recovery [%]) and in vitro properties of stored red cells (RBCs). There is uncertainty as to how these changes are related to the day of irradiation and the length of storage after irradiation. STUDY DESIGN AND METHODS: Four protocols used day of irradiation and storage time after irradiation as the independent variables. At the conclusion of the storage period, viability was measured with radiolabeled RBCs as the recovery and the long-term survival time for RBCs that were circulating beyond 24 hours. In addition, in vitro values including RBC ATP, hemolysis level, and supernatant potassium were measured. Each subject donated 2 units of whole blood (CPD) and received autologous irradiated and untreated control RBCs (AS-1) on two separate occasions. RESULTS: Reduced recovery in irradiated units was noted when compared to that in control units, and the reduction was most apparent with long periods of storage after irradiation, irrespective of the day of irradiation. With irradiation on Day 1 of storage and a total storage period of 28 days, mean +/- SD recovery (single label) was 84.2 +/- 5.1 percent for control RBCs and 78.6 +/- 5.9 percent for irradiated RBCs (n = 16; p<0.01). With irradiation on Day 14 and storage through Day 42, the recoveries were 76.3 +/- 7.0 percent for control RBCs and 69.5 +/- 8.6 percent for irradiated RBCs (n = 16; p<0.01). Less reduction in recovery was observed with shortening of the postirradiation storage time. When the total storage period was reduced to 28 days after Day 14 irradiation, the recoveries were not significantly different. With an additional 2-day storage period after irradiation on Day 26, the recoveries were also comparable. Long-term survival times for control and irradiated RBCs were not significantly different in any of the four protocols. RBC ATP levels and hemolysis were minimally, but significantly influenced by irradiation. Supernatant potassium levels, however, were substantially increased after irradiation in each of the four protocols. CONCLUSION: Irradiation has only a small effect on the properties of RBCs treated and stored according to the utilized protocols. Longer storage times after irradiation resulted in progressively reduced recovery while long-term survival remained unaffected.     

Blood preservation XVI packed red cell storage in CPD-adenine

Interest has been renewed in CPD-adenine as a long-term liquid blood preservative. The question of whether the metabolic product of adenine, 2,8-dioxyadenine was toxic to humans has apparently been resolved by extensive animal and human studies in favor of there being no potential toxicity in the amounts used in blood preservation. Sweden is adopting CPD-adenine (0.25 mM) as its national blood preservative after ten years of clinical experience in trials. They have shown that each additional week of storage time beyond the current three weeks with CPD results in a 50 per cent reduction of wasteage caused by outdating. They are adopting the 35-day time for regular use with 42 days for an emergency reserve supply. However, many units of blood in the U.S. are stored as packed red blood cells and the question has been raised as to whether there is sufficient glucose in the preservative to maintain red blood cell metabolism in the packed cell unit. The present investigation indicates that there is sufficient glucose for 35 days of packed cell storage in CPD-adenine (0.25 mM) but in some units this might be marginal at 42 days of storage.     

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.     

Prestorage leucoreduction improves several in vitro red cell storage parameters following gamma irradiation

Gamma irradiation (GI) can prevent transfusion-associated graft-vs.-host disease, result in a small decrement in 24-h in vivo red blood cell (RBC) recovery and in a minor loss of maintenance of several in vitro parameters during routine storage. Results from studies by Davey et al. (Transfusion 1995, 35, 55S) demonstrated that the decrement in 24-h recovery was not observed if units were first leucoreduced (LR), suggesting that RBC damage on leucocytes from GI was secondary to the effects of irradiation . In this study, we further investigated how leucoreduction affected eight in vitro storage parameters of gamma-irradiated units by using a matched sample study design. Leucoreduction significantly reduced (P < 0.05) the deleterious effect of GI on adenosine triphosphate glucose levels, haemolysis and mean corpuscular volume (MCV) for AS-3 units and significantly decreased (P < 0.05) the deleterious effect of GI on glucose levels, haemolysis and MCV for AS-1 units. Leucoreduction did not influence (P > 0.5) the enhanced potassium release associated with GI. Some, but not all, the observed RBC damage caused by GI appears to be secondary to the effect of irradiation on leucocytes. Based on the in vivo survival studies of Davey et al. and the results of this study, there is justification to consider performing additional survival studies to support extended storage of LR gamma-irradiated RBC.     

Effects of red cell storage and lysis on in vitro cytokine release.

Cytokine release in whole blood assays is inhibited by addition of supernatants from stored blood components in a storage time dependent manner. This is also observed after prestorage leucofiltration of the blood. The role of erythrocyte changes during storage is unknown. In this study, we have therefore used prestorage leucofiltered buffy coat-depleted red blood cells stored for 35 days. Assays of whole blood with addition of either complete suspension or only supernatant from the stored blood units were stimulated with lipopolysaccharide (LPS) and incubated. Additionally, samples from the stored blood units were partly haemolysed and added to similar whole blood assays. After incubation, tumour necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) concentrations were determined. Results showed that the TNF-alpha release was not changed by addition of supernatants from prestorage leucofiltered blood, but significantly decreased with addition of red cell suspension in a storage time dependent manner. The IL-10 release increased with storage time with addition of both red cell suspension and supernatant. The TNF-alpha/IL-10 ratio decreased with storage time, but significantly more, 25% vs. 10%, during the 35 days, with addition of suspension compared with addition of supernatant. TNF-alpha and IL-10 release was not changed by addition of supernatant from lysed erythrocytes of various storage times. Therefore, storage time dependent inhibition of immune response by red cell suspension may in part be dependent on red cell presentation, and may cause some of the side effects by transfusion of red cells.     

低功率半导体激光照射对体外培养嗅鞘细胞的影响

目的研究低功率810 nm半导体激光对体外培养嗅黏膜嗅鞘细胞增殖的影响。 方法取成年SD大鼠嗅黏膜,采用差速贴壁法获得嗅鞘细胞,对细胞进行不同功率密度的半导体激光照射,波长为810 nm,在10.3 mW/cm2功率密度下分别照射30,60,120 s,每日1次,连续3 d,各次间隔24 h。最后一次照射后细胞培养。主细胞培养第3,5,8天时采用细胞计数法和四氮唑蓝（MTT）法分别检测照射后不同时间细胞增殖的变化。 结果最后一次照射后细胞培养第3天时,不同时间照射组和对照组细胞增殖无差异。细胞培养第5天和第8天时,不同时间照射组细胞数均高于对照组,差异有统计学意义（P＜0.05）;60 s和120 s照射组OD值高于对照组,差异有统计学意义（P＜0.05）;但30 s照射组OD值与对照组差异无统计学意义。其中以60 s照射组细胞增殖最为明显。 结论低功率半导体激光对体外培养嗅黏膜嗅鞘细胞的增殖有促进作用,其作用与照射的时间剂量有关。最佳照射时间是每日1次,连续3 d,各次间隔24 h。     

Effects of gamma irradiation on red cells from donors with sickle cell trait

BACKGROUND: Transfusion-associated graft-versus-host disease can be prevented by gamma irradiation of blood components. Red cells (RBCs) from sickle cell disease patients may exhibit oxidative changes of RBC membranes due to the instability of hemoglobin (Hb) S. Persons with sickle cell trait are eligible to donate blood, and 35 to 45 percent of their total Hb is Hb S. The effect of gamma irradiation on RBCs from such persons is of interest. STUDY DESIGN AND METHODS: RBCs from 12 donors with sickle cell trait (Hb AS) and from 12 with normal Hb (Hb AA) were studied. Each of the 24 RBC units was divided equally into two transfer bags via a sterile connecting device. One bag from each RBC unit received a 2500-cGy dose of gamma irradiation at its mid-plane and was stored at 4 degrees C; the second set of bags was stored without irradiation. For RBCs from 6 donors with Hb AS and 6 donors with Hb AA, units were irradiated on Day 7 and studied on Day 35 of storage (Group 1). For the RBCs from the other 6 donors with Hb AS and the other 6 donors with Hb AA, units were irradiated on Day 28 and studied on Day 42 of storage (Group 2). RESULTS: For Group 1 and Group 2, plasma potassium and plasma Hb concentrations were significantly higher and RBC ATP concentrations were slightly lower in the irradiated units than in the nonirradiated units. In Group 1 and Group 2, there were no significant differences in the plasma potassium or RBC ATP concentrations in either the irradiated or the nonirradiated units of RBCs from donors with Hb AS and donors with Hb AA. Plasma Hb concentrations were consistently lower in the units from donors with Hb AS, whether or not they were irradiated. However, in both groups, proportionally similar changes in plasma Hb concentration were detected when the irradiated Hb AS and Hb AA units were compared to nonirradiated Hb AS and Hb AA units. CONCLUSION: Gamma irradiation of RBCs from donors with Hb AS or with Hb AA resulted in comparable changes in plasma potassium, RBC ATP, and plasma Hb concentrations, although donors with Hb AS had lower plasma Hb. RBCs from donors with Hb AS subjected to 2500 cGy of gamma irradiation did not evidence a storage lesion greater than that seen in RBCs from donors with Hb AA.     

Length of storage and in vitro immunomodulation induced by prestorage leukoreduced red blood cells

BACKGROUND: The relationship between length of storage of red blood cell (RBC) units and biochemical changes has been well studied, but little is known about the progression of cellular immunomodulative properties in blood recipients. This study aims to quantify in vitro T-cell activation and cytokine release by white blood cells, after incubation with supernatants from leukoreduced RBCs.
STUDY DESIGN AND METHODS: Whole blood cultures were incubated with supernatant from five leukoreduced RBC units stored for 1, 6, 10, 15, 24, and 42 days. Supernatant-induced T-cell activation was evaluated by quantifying CD25 expression. Supernatant-induced cytokine production was determined by measuring interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α levels.
RESULTS: No cytokines were detected in RBC supernatants even after 42 days of storage. However, IL-6 levels in whole blood culture increased significantly when incubated with supernatant from RBC units stored for 1, 6, and 15 days, by factors of 1.7 ± 0.3, 1.7 ± 0.3, and 1.4 ± 0.3, respectively. TNF-α levels were significantly decreased on Days 24 and 42 of storage by factors of 0.50 ± 0.42 and 0.33 ± 0.21, respectively. IL-10 levels were significantly increased on Days 1 and 42 of storage by factors of 2.3 ± 1.3 and 3.2 ± 2.8, respectively. After an initial increase in IL-6 and TNF-α production, there was a significant linear decrease in their levels measured from units stored for longer times. No significant changes in CD25 expression were observed over time.
CONCLUSION: Although no cytokines were measured in the supernatants from leukoreduced RBCs, these supernatants exhibited variable immunomodulatory effects related to their length of storage.     

胶体[^32P]对原代培养大鼠前列腺细胞凋亡的研究

目的观察不同浓度胶体[^32P]对原代培养大鼠前列腺细胞凋亡的影响。方法原代培养大鼠前列腺细胞,建立辐射诱导前列腺细胞凋亡模型,采用吖啶橙染色和流式细胞术检测大鼠前列腺细胞在不同辐射剂量下的细胞凋亡变化。结果对照组前列腺细胞呈典型的上皮型细胞形态,小剂量辐射组未见明显改变,大剂量辐射组可见有散在的细胞核固缩、碎裂。荧光显微镜下,小剂量辐射组与对照组细胞胞核DNA呈黄色均匀荧光,大剂量辐射组前列腺凋亡细胞的胞核或胞质内可见致密浓染的黄绿色荧光。小剂量辐射组与对照组细胞凋亡率均很低,大剂量辐射组细胞凋亡率明显增多（P〈0．01）,并可见亚二倍体峰。结论低剂量辐射对前列腺上皮细胞的致死性损伤作用不大,仅能引起少量细胞凋亡,大剂量辐射可致大量前列腺细胞发生凋亡。     

Preparation of packed red cells suitable for intravascular transfusion in utero

Intra-cardiac transfusion in utero is a recent development in the treatment of erythroblastosis foetalis. As with intra-umbilical cord transfusion, the procedure has been described as having a high risk of complication characterized by bradycardia and cardiac-arrest. Intra-coronary hyperkalemia was suspected as being a major contributing factor towards the complications experienced, particularly by the intra-cardiac delivery. The procedure is warranted, however, in cases where intra-umbilical cord transfusion is not possible. Therefore, studies comparing various methods of preparing packed cells for intravascular transfusion were initiated. The plasma potassium levels of gravity sedimented packed cells and saline washed red cells resuspended in either Ringer's lactate, pooled donor plasma or saline were measured. Products of saline washed/resuspended packed red cells transfused within 2 hours of preparation were found to have low potassium levels while exposing the fetus to minimal risk of transfusion transmitted infection and were therefore selected to be the most suitable product. Using this product, a significant decrease in the incidence of severe bradycardia resulting in cardiac-arrest, following intravascular transfusion (32 procedures) was observed.     

Microaggregate content and flow rates of packed red blood cells

Platelet rich red cells (PRRC) and platelet poor red cells (PPRC) are both prepared in the course of red cell production. PRRC tend to have a higher hematocrit (82 +/− 7) than PPRC (77 +/− 5), (p less than .001). There are more microaggregates present in PRRC (2.48 +/− 1.41 gm) than in PPRC (1.46 +/− 0.61 gm), (p less than .001). The infusion rate for PRRC was 4.0 × 1.8 ml/minute, and this was significantly smaller than for PPRC which was 9.7 +/− 1.7 m/minute, (p less than .001). This compares to a rate for whole blood of 32.8 ml/minute. The difference in flow rate of the two types of red blood cells is in part due to a difference in viscosity, but more inportantly due to a difference in microaggregate content. Flow rate is normalized for both types of packed cells by the addition of 100 to 150 ml of saline, while infusion time is normalized by the addition of only 50 to 100 ml of saline to the packed cell units.     

Circulatory risk in the transfusion of red blood cells with impaired flow properties induced by storage

Red blood cell (RBC) flow properties (FPs), specifically their deformability, aggregability, and adherence to endothelial cells, play major roles in blood circulation. Their impairment, as occurs under various blood banking conditions, may contribute to circulatory impairment in recipients. Recent studies and meta-analyses show that the transfusion of stored RBCs (stRBCs) may be less beneficial than that of freshly collected units, which may thus adversely affect recipients, especially their circulatory function, thereby pointing to a potential role in the alteration of FPs of stRBCs. In this review, we present an up-to-date summary of the studies on the FP of stRBCs, clearly showing that they may be impaired at an early stage of storage, which may contribute considerably to transfusion-associated circulatory impairment in recipients. The alteration of the FPs of stRBC is attenuated by prestorage leukoreduction and/or poststorage "rejuvenation." However, because these procedures, especially rejuvenation, are costly and are associated with an increased risk of bacterial contamination, there is an urgent need to establish better methods of improving the hemodynamic function of stRBCs before their transfusion. It is therefore proposed that the FPs of stRBC may be important measures of the outcome of RBC transfusions. Monitoring such functions would thereby introduce necessary criteria and new tools for the quality control of stRBC units, making an important contribution to transfusion therapy.     

白细胞滤除对保存期内红细胞流变性和形态的影响

目的 探讨白细胞滤除对保存期红细胞流变性及形态的影响.方法 选择30名健康献血者的血液制备成红细胞悬液,随机分为实验组(n=30):使用去白细胞输血过滤器去除红细胞悬液中的白细胞(简称滤白组);对照组(n=30):未滤白的红细胞悬液;2组一起常规保存.取采血后d0、d7、d14、d21、d28、d35的血标本作白细胞(WBC)和红细胞计数(RBC)、红细胞压积、血液高剪切力、低剪切力及细胞形态学检测.结果 过滤前后红细胞悬液内的WBC为(6.80±0.85)(× 109/L) vs (3.12±0.26)(×106/L) (P ＜0.01);保存d21时低、高切粘度分别为:对照组(11.28±1.88)1/s、(2.85±0.29)200/s,滤白组(12.36±1.57)1/s、(2.93±0.22) 200/s,较保存1～2周明显上升(P＜0.05),但组间比较未见明显著变化(P＞0.05);瑞氏染色结果显示2组细胞形态也有不同变化,滤白组红细胞形态保存较好.结论 白细胞滤除能有效减少白细胞崩解产物或分泌因子对红细胞形态的影响.     

储存前去白细胞悬浮红细胞在保存期内质量变化的研究

目的 探讨储存前去白细胞悬浮红细胞在保存期内质量变化的研究.方法 选择20名符合《献血者健康检查要求》的献血者所献的400 mL全血,在24h内将其分成2份200 mL全血,将其中1份制备成红细胞悬液为对照组(n=20);另1份使用白细胞滤器去除白细胞后再制备成去白红细胞悬液为实验组(n=20),2组一起4±2℃保存.取采血后1d、7d、14 d、21 d、28 d、35 d对血标本作血常规(PBC、HGB、Hct、MCV)、生化(K+、Na+、Cl-)、血液流变学(全血粘度、全血还原粘度、红细胞聚集指数、红细胞刚性指数、红细胞变形指数、红细胞电泳指数)、红细胞渗透脆性、游离血红蛋白和储存期末溶血率检测.同时分别对2组数据进行统计学分析.结果 去白红细胞悬液组和红细胞悬液组的K+、游离血红蛋白和储存期末溶血率随着保存时间的延长而有所升高,2组数据在相同储存时间差别不显著(P＞0.05),均符合悬浮红细胞质量国家标准.红细胞聚集指数、红细胞刚性指数和红细胞电泳指数、全血粘度随着保存时间的延长而有所升高,在保存14 d后相同时间差别显著(P＜0.05),红细胞悬液组高于去白红细胞悬液组.全血还原粘度随着保存时间的延长而有所升高,在保存28 d后相同时间差别显著(P＜0.05),红细胞悬液组高于去白红细胞悬液组.其它指标变化不明显.结论 储存前去白细胞悬浮红细胞不仅白细胞滤除,红细胞的血流变学指标比未滤除的红悬液好,所以建议储存前滤白红悬液更能保证血液质量与安全.     

红光照射对战士运动前后肺功能的影响

目的 观察经红光照射后战士在运动后肺功能的影响.方法 2007-03～2007-04,对20名健康的战士进行红光照射,照射1次/d,照射时间为0.5 h/次,在照射前、照射1周和2周后分别进行长跑运动前后的肺通气功能测试及采静脉血测定生理指标:白细胞计数(WBC)、红细胞计数(RBC)、血红蛋白(Hb)、丙氨酸氨基转移酶(ALT)、门冬氨酸转移酶(AST)、肌酸激酶同功酶(CK-MB).结果 运动前红光照射2周后的肺活量(VC)、最大通气量(MVV)、用力呼气1 s量(FEV1)、用力呼气1 s率(FEV1%)及中期流速(MMF)与照射前相比都有明显增加(P<0.05,P<0.01).运动后FEV1%和MMF在照射2周与照射前相比有明显增加(P<0.05).照射1周后的Hb、照射2周后的RBC与照射前相比都有明显增加(P<0.05);照射2周后的Hb有显著增加(P<0.01).生化指标中各种酶均下降,其中CK-MB值下降显著(P<0.05).结论 红光能促进机体在运动时维持身体的正常机能,延缓运动疲劳的发生,提高运动耐力.