术中患者回收血与库存血红细胞携氧能力及平均年龄的比较

目的比较患者术中回收血红细胞与库存血红细胞携氧能力及平均年龄的差异。方法使用血液回收机对30名手术患者行术中自体血液回收,取血液回收机回收处理后的红细胞和库存异体去白细胞悬浮浓缩红细胞各30份,检测2组血样的pH、氧分压及血氧饱和度,计算血样的P50,采用酶联免疫法检测红细胞内2,3-二磷酯甘油酸浓度、化学法检测红细胞丙酮酸激酶活性。结果回收血组的P50明显低于库存血组;回收血组的pH值、2,3-二磷酯甘油酸浓度、丙酮酸激酶活性均明显高于库存血组,差异有统计学意义(P<0.05)。结论回收血红细胞携氧能力优于库存血,回收血红细胞平均年龄较库存血更为年轻。     

血液回收对红细胞ATP含量及葡萄糖6-磷酸脱氢酶活性的影响

目的探讨术中自体血液回收对红细胞ATP含量及葡萄糖6磷酸脱氢酶(G6PD)活性的影响。方法选择心血管外科术中出血量>600ml患者50名,用血液回收机行自体血液回收,取患者术野及回收的红细胞各6ml,另取50份库存2周的异体红细胞悬液6ml,分别检测红细胞ATP含量及G6PD活性。结果回收组和库存组ATP含量、G6PD活性与术野组相比均明显下降(P<0.01),但仍在正常值范围;回收组ATP含量及G6PD活性与库存组相比,差异无显著性。结论血液回收处理可使红细胞ATP含量减少,使G6PD活性降低,但未达影响红细胞功能的程度,回收红细胞与库存红细胞相比差别不明显。     

术中洗涤式回收自体血与库存血红细胞携氧能力的比较

目的:比较洗涤式回收自体血红细胞与库存异体血红细胞携氧能力,探讨两种临床用血的差异,指导临床输血。方法:回收血组:采用国产自体-3000型血液回收机留取术中自体血液;库存血组:取库存9~15 d的血液。用血气分析仪检测两组血样的细胞内钠(Na+)、钾(K+)、pH值、细胞携氧能力(用P50表示使Hb氧饱和度达到50%的氧分压)。结果:回收血组红细胞内pH、Na+值高于库存血组(P0.05);红细胞内K+浓度低于库存血组(P0.05);红细胞P50明显低于库存血组(P0.05)。结论:术中回收自体血的红细胞携氧能力优于库存异体血红细胞。     

骨科手术中回收血红细胞与患者麻醉前红细胞携氧能力的比较

[目的]比较骨科患者术中回收血红细胞与麻醉前红细胞携氧能力的差异.[方法]使用血液回收机对30例骨科手术患者行术中自体血液回收.取血液回收机处理后的回收血和麻醉前患者外周血各30份,检测两组血样的pH、根据公式计算血样的50%氧分压(P50),采用放射免疫法测定红细胞内2,3-二磷酸甘油酸浓度(2,3-DPG)、在血液...     

自体回收血与库血红细胞酶活性及ATP含量的对比

目的比较术中自体血回收处理和库存2周红细胞的酶活性以及红细胞ATP含量的差异。方法选择估计术中出血量在800 ml以上的择期手术患者50例,术中用自体血液回收机进行洗涤血液回收,取患者自体回收处理血及库存2周的异体浓缩红细胞各50份,每份6 ml,分别检测红细胞ATP酶（Na＋-K＋-ATP酶、Ca2＋-ATP酶、Mg2＋-ATP酶）,葡萄糖-6-磷酸脱氢酶（G-6PD）活性及红细胞ATP的含量。结果红细胞ATP酶的活性：回收血组Na＋-K＋-ATP酶、Ca2＋-ATP酶、Mg2＋-ATP酶的活性与库血组各对应的ATP酶活性相比,差异无统计学意义。红细胞ATP含量与G-6PD活性：回收血组红细胞ATP含量、G-6PD活性与库血组相比,差异无统计学意义。结论自体血液回收处理的红细胞ATP酶,G-6PD活性及ATP含量与库存2周异体红细胞间差异均无统计学意义。     

骨科患者术中回收血红细胞携氧能力的研究

目的 比较骨科患者术中回收血红细胞与麻醉前红细胞携氧能力的差异.方法 使用血液回收机对40名骨科手术患者行术中自体血液回收,取血液回收机处理后的回收血和麻醉前患者外周血各40份,检测两组血样的pH、计算血样的50％氧分压(P50),采用放射免疫法测定红细胞内2,3-二磷酸甘油酸浓度(2,3-DPG)、在血液流变仪上测定红细胞变形指数(RCD).结果 回收血组pH值、P50、红细胞内2,3-DPG、RCD与麻醉前外周血组比较差异均无统计学意义(P＞0.05).结论 骨科患者术中回收血与麻醉前外周血红细胞具有相似的携氧能力.     

颅脑手术中清洗后回收血液质量的研究

目的检验颅脑手术中血液回收机处理后回收血液的质量。方法20名颅脑疾患择期手术患者,应用自体-2000型血液回收机回收术野出血,经抗凝、过滤、离心及清洗后回输。测定血液回收机对收集血的处理过程中血小板、白细胞、K+和游离血红蛋白的清除率。用扫描电子显微镜对5份清洗后的血样及5U库存2周的红细胞悬液进行超微形态学检验,观察红细胞的形态。结果储血罐内收集血的溶血率达到(8.57±0.35)%,血液回收机的离心清洗过程中血小板、白细胞、K+和游离血红蛋白的清除率分别为:(85.4±7.3)%、(38.8±17.1)%、(90.4±4.4)%及(92.7±2.9)%。扫描电子显微镜下虽然可见回收血液中含有部分变形的红细胞,但红细胞的形态正常率仍达到(58.0±8.0)%。而库存2周的红细胞悬液中含有更多变形的红细胞(P<0.01)。结论在颅脑手术中收集的血液,溶血程度较重,但经血液回收机处理后,其质量较好。     

红细胞膜骨架收缩蛋白及其基因突变

血影收缩蛋白通过自身聚合由异二聚体形成四聚体（ＳＰＴ），参与构成红细胞膜蛋白质网状骨架，并与膜骨架蛋白其它成分及膜固有蛋白质相互作用，维持正常红细胞的形态和功能。收缩蛋白基因突变导致红细胞收缩蛋白自身聚合位点及其它结构区域的异常，使ＳＰＴ形成障碍，均使红细胞膜结构与功能出现异常。     

清洗式自体回收血液的红细胞特点

背景：清洗式自体血液回输是将术野血经回收、抗凝、过滤、离心、浓缩、清洗后回输给患者的自体血液回输方式,在临床中已大量应用。
　　目的：归纳总结清洗式自体回收血液的红细胞特点,包括红细胞的回收率和红细胞压积,红细胞的形状、变形能力、血流动力学和体内生存期的变化,红细胞携氧及供氧能力的变化以及红细胞免疫以及其表面膜受体的变化。
　　方法：由第一作者检索1987年1月至2013年1月 PubMed数据及万方数据库相关文献,英文检索词为“Blood transfusion,autologus,blood preservation,erythrocytes”,中文检索词为“输血;自体;血液保存;红细胞”。计算机初检得到与清洗式自体回收血液的红细胞特点相关文献200篇文献,排除重复性研究,保留60篇做进一步总结分析。
　　结果与结论：由于受负压吸引、离心分离等机械力、各种受损组织及细胞释放的炎性递质和酶类以及激活的补体等因素的共同作用,回收红细胞有一定程度的破坏,所以血液回收机对红细胞的总回收率取决于采集时的回收率、贮存破损率和清洗时的丢失率。自体血液回收对红细胞的携氧能力无明显影响,即回收血红细胞具有与正常红细胞相同的携氧能力。红细胞在自身输血前后免疫功能和表面受体的数量有所下降,但优于库存2周的红细胞。研究提示应提高血液回收技术以降低红细胞免疫黏附功能下降程度。     

库存血红细胞表面C3b、G6PD水平变化调查

目的 观察库存血有效期内红细胞表面C3b、G6PD水平变化过程,了解保存期库存血红细胞免疫功能及细胞膜的微观变化.方法 红细胞表面C3b采用卡式微柱凝胶法检测;G6PD采用自动生化分析仪检测.结果 陈旧库存血（采血30 d后）与新鲜库存血（采血7d内）比较,红细胞表面C3b、G6PD水平均显著下降.结论 有效期内库存血液保存时间越长,红细胞免疫功能越低,G6PD活性也相应减弱.这种变化应引起输血界重视.     

A single assay for multiple storage‐sensitive red blood cell characteristics by means of infrared spectroscopy

BACKGROUND: To maintain a high quality of red blood cells (RBCs), RBC characteristics must be followed during storage under blood bank conditions. By means of infrared (IR) spectroscopy, several characteristics can be measured simultaneously. STUDY DESIGN AND METHODS: IR spectra were acquired for samples from RBCs that were collected and stored according to Dutch blood bank procedures for a period of up to 50 days. Spectra of the soluble cell components were acquired separately after hypotonic lysis of the cells, followed by centrifugation. Characteristic vibrational bands were analyzed with respect to storage time–dependent changes in peak position and in intensity. RESULTS: A decrease in corresponding peak intensities indicates that RBCs lose protein and lipid during storage. Changes in protein secondary structure during storage are largely confined to integral membrane proteins and membrane‐associated proteins. A concurrent decrease in lipid packing density probably reflects the gradual change in cellular shape from discoidal to globular. By integration over a narrow range, storage‐dependent changes in intracellular adenosine triphosphate (ATP) and glucose levels could be estimated. ATP levels decrease during storage, but stay above the required 75% of the initial level after 35 days of storage. Glucose concentrations stay well above 5 mmol/L over the entire storage period. CONCLUSION: IR spectroscopy is a promising technique to follow structural and metabolic changes in RBCs during storage under blood bank conditions. Several variables can be determined rapidly in a single measurement.     

Decreased immunorejection in unmatched blood transfusions by attachment of methoxypolyethylene glycol on human red blood cells and the effect on D antigen

BACKGROUND: Pegylation of red blood cells (RBCs) has been the primary focus of research on the immunocamouflage of cell. The aim of this study was to demonstrate pegylation homogeneity, its shielding effect on D antigens, and its storage stability. In addition, methoxypolyethylene glycol (mPEG)-modified RBCs (mPEG-RBCs) were tested serologically against a panel of serum samples that was difficult to match to find a solution to the difficulty in matching. STUDY DESIGN AND METHODS: In this study, fluorescein-PEG and a confocal laser scanning microscope were used to monitor PEG attachment on RBC population and observe reaction homogeneity, the stability of mPEG combined with RBCs in vitro was evaluated by the RBC ghost agglutination test, the pegylation sites on membrane were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with two dye methods, and the effect of pegylation on D antigen was detected by immunoblotting techniques. Compatibility tests were carried out between 66 cases of serum with difficulty in blood matching and mPEG-camouflaged RBCs by use of four blood matching methods including direct agglutination, indirect antiglobulin test (IAT), microtyping gel cards (MTS), and the manual polybrene technique (MPT). RESULTS: The results indicated the homogeneity of pegylation, the absence of RhD protein in mPEG-modified D+ RBCs by Western blotting, and attachment of PEG to RBCs after 30 days of storage, while RBCs still remained antigenically silent. All pegylation RBCs showed a negative reaction with ABO-matched patients' serum samples by direct agglutination, IAT, and MTS, which indicated that pegylation RBCs and patients' serum samples were compatible. MPT was not suitable for detecting blood matching of mPEG-RBCs, because modification changed the RBCs' biophysical properties. CONCLUSION: In conclusion, mPEG-RBCs have acceptable in vitro properties and provide a useful solution to problems with clinical blood matching, although such masking leaves much to be desired.     

Interleukin-11 therapy selectively downregulates type I cytokine proinflammatory pathways in psoriasis lesions

Protein 4.2 is a major component of the red blood cell (RBC) membrane skeleton. We used targeted mutagenesis in embryonic stem (ES) cells to elucidate protein 4.2 functions in vivo. Protein 4. 2-null (4.2(-/-)) mice have mild hereditary spherocytosis (HS). Scanning electron microscopy and ektacytometry confirm loss of membrane surface in 4.2(-/-) RBCs. The membrane skeleton architecture is intact, and the spectrin and ankyrin content of 4. 2(-/-) RBCs are normal. Band 3 and band 3-mediated anion transport are decreased. Protein 4.2(-/-) RBCs show altered cation content (increased K+/decreased Na+)resulting in dehydration. The passive Na+ permeability and the activities of the Na-K-2Cl and K-Cl cotransporters, the Na/H exchanger, and the Gardos channel in 4. 2(-/-) RBCs are significantly increased. Protein 4.2(-/-) RBCs demonstrate an abnormal regulation of cation transport by cell volume. Cell shrinkage induces a greater activation of Na/H exchange and Na-K-2Cl cotransport in 4.2(-/-) RBCs compared with controls. The increased passive Na+ permeability of 4.2(-/-) RBCs is also dependent on cell shrinkage. We conclude that protein 4.2 is important in the maintenance of normal surface area in RBCs and for normal RBC cation transport.     

The experience of flow cytometry for specific antibody against cisplatin-treated red blood cells: A case report

Background

Cisplatin-associated hemolysis is a rare but important adverse effect. Nonimmunological protein adsorption (NIPA) due to erythrocyte membrane modification has been reported as the leading cause of cisplatin-associated hemolysis. However, limited data exist on cisplatin-associated immunological hemolysis because of a lack of an established diagnostic method. Here, we used flow cytometry (FCM) to diagnose a patient with cisplatin-associated immunological hemolysis.

Study Design and Methods

A 55-year-old woman with uterocervical cancer was treated with weekly cisplatin monotherapy (40 mg/m²). She had no previous transfusion and medication history, nor any significant family history. On the 26th day after cisplatin administration, severe hemolysis was noted. Her red blood cells (RBCs) and sera were evaluated by direct antiglobulin test (DAT) and indirect antiglobulin test (IAT), respectively. To explore immunological reactions for cisplatin-treated RBCs, we attempted FCM using cisplatin-treated and -untreated RBCs. After incubating conditioned RBCs with the patient's serum or healthy donor serum, we evaluated their fluorescent intensity by fluorescein isothiocyanate (FITC)-conjugated anti-human immunoglobulin (Ig) G antibodies.

Results

The patient's DAT was positive, and an IAT using her plasma was positive for cisplatin-treated RBCs. FCM using cisplatin-treated RBCs revealed that the patient's serum had higher FITC intensity than the donor's serum, indicating the existence of cisplatin-treated RBC-specific IgGs in patient's serum.

Conclusion

Here, we report a rare case of a patient with hemolysis diagnosed using FCM to identify specific antibodies against cisplatin-treated RBCs. NIPA and immunological mechanisms may contribute to hemolysis onset during cisplatin treatment.     

三种试剂对红细胞表面CD38抗原去除效果的研究

目的比较0.2 mol/L二硫苏糖醇(DTT)、0.2 mol/L二巯基乙醇(2-ME)及1%菠萝蛋白酶对红细胞表面CD38抗原去除的效果。方法将2%~5%红细胞悬液与0.2 mol/L DTT、0.2 mol/L 2-ME及1%菠萝蛋白酶按照体积比1∶4分别混匀,37℃孵育30 min洗涤后制备试验用红细胞悬液,以未经处理的红细胞悬液作为阳性对照。上述试验用红细胞悬液与含有达雷妥尤单抗(抗-CD38)的患者血浆进行间接抗人球蛋白试验(IAT)。结果经0.2 mol/L DTT、0.2 mol/L 2-ME及1%菠萝蛋白酶处理过的红细胞与患者血浆进行IAT,前二者处理后结果为阴性,后者显微镜下观察为弱阳性。阳性对照结果为1+。进行IAT期间发现处理过的红细胞均发生不同程度的溶血现象,最严重的为1%菠萝蛋白酶处理过的红细胞。结论上述三种试剂均能去除红细胞表面CD38抗原,但0.2 mol/L DTT、0.2 mol/L 2-ME去除效果优于1%菠萝蛋白酶。     

Red blood cell storage and cell morphology

Aim: In this study, we performed weekly assessment of morphology‐related parameters through monitoring of CPD‐SAGM leuco‐filtered erythrocyte concentrates from blood withdrawal until the 42nd day of storage. Background: Liquid storage of red blood cells (RBCs) delivers a blood‐derived therapeutic, which is safe, available, effective and affordable for most patients who need transfusion therapy in developed countries. However, a growing body of accumulating controversial evidences, from either biochemical or retrospective clinical studies, prompted safety concerns about longer stored RBCs. Methods: Statistical image analysis through scanning electron microscope was coupled to osmotic fragility and erythrocyte sedimentation rate. Results: We could observe that by day 21 more than 50% of RBCs displayed non‐discocyte phenotypes. This observation was related to an increase in osmotic fragility, which was totally overlapped in day 0 controls and day 7 RBCs while only slightly augmented in day 14 samples. Cation dysregulation (pH internal/external alteration and potassium) might both reflect and trigger a negative feedback loop with metabolic fluxes and membrane cation pumps. Conclusion: Morphology parameters suggest that significant alterations to RBC morphology over storage duration occur soon after the 14th day of storage, as to become significant enough within the 21st day.     

Red blood cell calcium homeostasis in patients with end-stage renal disease

Low cell calcium level is essential for preservation of red blood cell (RBC) membrane deformability and survival. RBCs from patients with end-stage renal disease (ESRD) demonstrate reduction in membrane deformability, possibly as a result of increased RBC cellular calcium level. To evaluate calcium homeostasis in RBCs from patients with ESRD, we measured cell calcium level, basal and "calmodulin"-stimulated calcium-stimulated Mg-dependent ATPase (CaATPase) activity, and calcium 45 efflux were measured before and after hemodialysis. The in vitro effect of uremic plasma and of urea on CaATPase activity of normal RBCs was tested, and 45Ca influx into RBCs of patients undergoing hemodialysis also was determined. A morphologic evaluation of red cells from patients with ESRD was performed with a scanning electron microscope. RBC calcium level in patients (mean +/- SEM 21.2 +/- 2.8 mumol/L of cells; n = 28) was higher than in controls (4.9 +/- 0.3 mumol/L of cells; n = 24; p less than 0.001). Hemodialysis had no effect on cell calcium level. Both basal and "calmodulin"-stimulated RBC CaATPase activities in patients with ESRD (n = 9) were reduced by approximately 50% (p less than 0.01), but after hemodialysis, enzyme activity returned to normal. 45Ca efflux from calcium-loaded cells, which was 2574.0 +/- 217.0 mumol/L of cells per 0.5 hours before hemodialysis, increased to 3140.7 +/- 206.8 mumol/L of cells per 0.5 hours after hemodialysis (p less than 0.005). In vitro incubation of normal RBCs with uremic plasma depressed CaATPase activity, but incubation with urea had no effect. RBCs of patients with ESRD revealed increased 45Ca influx, 7.63 +/- 1.15 mumol/L of cells per hour versus 4.61 +/- 0.39 mumol/L of cells per hour (p less than 0.025). RBCs of patients revealed a high incidence of spherocytosis and echynocytosis, which correlated with a high cell calcium level (r = 0.894, p less than 0.01). These results indicate that RBC calcium level is elevated in patients with ESRD and suggest that a dialyzable uremic factor inhibits RBC CaATPase activity and thereby calcium efflux, which may account for the elevated cell calcium level. The increased calcium influx further increases cellular calcium level. These abnormalities are associated with spherocytosis and echynocytosis and may contribute to the shortened survival of RBCs in uremia.     

Detection of anti-A in neonatal serum

The thirteenth edition of the standards of the American Association of Blood Banks did not require the use of A1 red cells (RBCs) or an indirect antiglobulin test (IAT) to detect anti-A in neonatal serum, whereas the fourteenth edition mandates both. The present study was conducted to help document the need for these changes. Incomplete expression of the A antigen in neonatal patients can contribute to the accumulation of unabsorbed maternal anti-A that is capable of mediating the immune destruction of transfused adult RBCs. Sera from 50 group A neonatal infants of group O mothers were tested for anti-A by using RBC segments of A1 and A2 units of AS-1 RBCs less than 14 days old and also with A1 reagent RBCs. Of 22 sera in which anti-A was detected by RBCs from the A1 units with an IAT, anti-A was detected by RBCs from the A2 unit in only 1 and by the reagent RBCs in 17. In 19 (86%) of the 22 neonatal patients in whose sera anti-A was detected, the antibody was found only with the use of anti-human globulin in the IAT. It is concluded that testing to detect circulating anti-A in neonatal patients should include an IAT, and that it is preferable to use A1 RBCs for the initial evaluation.     

Evidence that CDw108 membrane protein bears the JMH blood group antigen

BACKGROUND: CDw108 is a cluster-of-differentiation antigen that resides on a glycosylphosphatidylinositol (GPI)-linked protein; it has not previously been shown to be expressed on red cells. JMH is a high- frequency red cell blood group antigen that resides on a GPI-linked protein of molecular weight similar to that bearing CDw108. The purpose of this study was to investigate whether CDw108 is expressed on red cells and whether it resides on the same membrane protein as does JMH. STUDY DESIGN AND METHODS: Murine monoclonal antibodies to CDw108, MEM- 121 and MEM-150, as well as a murine monoclonal antibody and human antibodies to JMH were used in radioimmunoassay, inhibition assay, Western blotting, and monoclonal antibody-specific immobilization of erythrocyte antigen assay. RESULTS: MEM-121 and MEM-150 were found to bind to red cells, and MEM-150 blocked binding of human anti-JMH to red cells. Anti-CDw108 and anti-JMH identified red cell membrane proteins that were of similar size and that were absent from JMH-negative red cells on Western blotting. MEM-150 and MEM-121 also immobilized the same protein that reacted with human anti-JMH. CONCLUSION: CDw108 is expressed on red cells and resides on the same GPI-linked membrane protein as does the JMH blood group antigen.     

Efficacy of intraoperative reservation of auto-blood in surgical treatment of ischemic heart disease using Cell-Saver

A prospective randomized study was made for 109 patients with ischemic heart disease (IHD) after coronary artery bypass grafting (CABG) with artificial blood circulation. Fifty-five (55) patients (Group 1--controls) were operated on without intraoperative autoblood reservation. Blood was sampled (7.6 +/- 0.3 ml/kg) at the surgery beginning in 54 patients (Group 2) with its subsequent transfusion after the artificial blood circulation was over. Cell saver was used in all patients intraoperatively (and, depending on indications--postoperatively). The dynamic content of hemoglobin, counts of erythrocytes, hematocrit parameters as well as the content of blood platelets and the content of blood-plasma total protein were examined postoperatively. The intraoperative autoblood reservation (7-8 ml/kg) made in CABG with cell saver for IHD patients maintains higher values of hemoglobin, erythrocyte counts and of hematocrit parameters during the early postoperative period, however, it does not reduce the frequency use or transfusion volume of donor blood.