12例急性自身免疫性溶血性贫血患者体外溶血试验配血与输血研究

本研究采用体外溶血试验配血方法,为微柱凝胶抗人球蛋白法配血困难的急性自身免疫性溶血性贫血（AIHA）患者筛查配血相容的红细胞。对照组选择26例AIHA患者,采用微柱凝胶抗人球蛋白法,确定配血结果凝集强度最弱的同型供血者红细胞输注。实验组为12例微柱凝胶抗人球蛋白法配血困难的急性AIHA患者,采用体外溶血试验配血法,选取配血结果出现不溶血的同型供血者红细胞输注。结果表明,体外溶血试验法输血组与微柱凝胶抗人球蛋白法配血组输血效果相比较,差异有显著性意义（P〈0.01）。实验组患者平均每次输注去白细胞红细胞悬液2.26单位,输血后患者血红蛋白、网织红细胞及总胆红素等指标变化与输血前相比较,差异有显著性意义（P〈0.01）。结论：在微柱凝胶抗人球法配血困难时,采取体外溶血试配血可以顺利为AIHA患者筛查到相容红细胞。     

溶血对血细胞计数影响的探讨

目的 探讨溶血对血细胞分析的影响.方法 选取门诊与住院患者37例（均无脂血、黄疸或溶血）,把无溶血标本在KX-21进行计数,记录结果作为无溶血结果,然后用注射器抽取混匀的抗凝血再用力打入试管3～5次,人工造成标本不同程度的溶血现象,再次混匀标本上机测定,作为溶血后的结果记录下来.用SPSS 15.0统计软件对溶血与不溶血结果作配对t检验.结果 淋巴细胞计数（LY）、单核细胞计数（MON）、红细胞计数（RBC）、红细胞平均压积体积（MCV）、平均红细胞血红蛋白浓度（MCHC）、平均红细胞血红蛋白含量（MCH）、红细胞分布宽度SD值（RDWSD）、血细胞压积（HCT）、血小板计数（PLT）、平均血小板压积（MPV）、血小板分布宽度（PDW）溶血前后比较差异均有统计学意义（P＜0.01）,白细胞计数（WBC）、中性分叶（Gron）、血红蛋白（Hb）溶血前后比较差异无统计学意义.结论 溶血会对血细胞计数产生严重干扰现象,通过简单离心能够观察标本是否溶血,严禁用溶血标本做血细胞计数.     

SPF级Balb/c小鼠脏器质量、脏器系数、血常规、血生化指标的测定与比较

背景：实验动物重要基础数据的标准化测定,是实行动物标准化监督管理的重要依据。目的：测定SPF级Balb/c小鼠主要脏器质量、脏器系数、血常规、血生化指标,并进行比较。方法：取18g左右Balb/c小鼠240只,雌雄各半,饲养1周后,精确称量体质量和主要脏器质量,计算脏器系数,并检测血常规及血生化指标。结果与结论：雌性与雄性Balb/c小鼠比较：①肺、脾、双肾、心、膀胱质量差异有显著性意义（P〈0.01或P〈0.05）。②肺、脾、双肾、膀胱脏器系数差异有显著性意义（P〈0.01或P〈0.05）。③白细胞、单核细胞、嗜碱性粒细胞百分比、红细胞、血红蛋白、红细胞平均血红蛋白浓度、红细胞分布宽度、嗜酸性粒细胞百分比差异有显著性意义（P〈0.01或P〈0.05）。④总蛋白、白蛋白、白球比、血糖、肌酐、尿酸、三酰甘油、总胆固醇、低密度脂蛋白、高密度脂蛋白、铁、磷、尿素氮差异有显著性意义（P〈0.01或P〈0.05）。说明性别对SPF级Balb/c小鼠主要脏器、主要脏器系数、血常规、血生化指标有影响。     

影响FQ—PCR检测HCVRNA结果的标本因素

目的：探讨脂血、溶血和胆红素对实时荧光定量PCR检测丙型肝炎病毒（HCV）RNA的影响。方法：根据不同要求收集特定的临床标本,采用荧光定量聚合酶链反应（FQ—PCR）测定标本中HCVRNA的含量。结果：高血脂组、溶血组和高胆红素组的检测结果与对照组比较差异均无统计学意义（P〉0．05）。结论：使用核酸提取柱法处理标本,可避免样本中脂血、溶血和胆红素等干扰物质对实验结果的影响。     

运动训练对大鼠脑缺血再灌注后功能恢复及VEGF表达的影响

目的：研究运动训练能否促进大鼠脑缺血再灌注后的功能恢复并探讨其机制.方法：30只SD大鼠随机分为对照组（n=10）、假手术组（n=10）和运动组（n=10）,制作大脑中动脉阻断（MCAO）再灌注模型,运动组再灌注24h起每天滚笼运动训练30min,分别在再灌注12h和21d测试行为功能,免疫组化法观察VEGF蛋白的表达.结果：再灌注12h,运动组和对照组的神经功能评分、前肢放置实验和后肢放置实验评分进行比较差异无显著性意义（P＞0.05）,再灌注21d,运动组行为功能评分优于对照组,比较差异有显著性意义（P＜0.05）;运动组再灌注12h VEGF蛋白的表达水平与对照组比较差异无显著性意义（P＞0.05）,再灌注21d VEGF蛋白表达水平运动组明显高于对照组（P＜0.05）.结论：运动训练促进大鼠脑缺血再灌注后的肢体功能恢复,其机制可能与VEGF表达上调有关.     

α-酮戊二酸脱氢酶在糖尿病骨骼肌病变中的作用

【目的】探讨STZ诱导的糖尿病骨骼肌病变大鼠模型中α酮戊二酸脱氢酶（α-KGDHC）活性及蛋白表达的变化。【方法】雄性Wister大鼠30只,随机分为实验组（n=15）和对照组（n=15）,两组大鼠分别于造模成功后第1月、3月、6月每组各取5只（实验Ⅰ、Ⅱ、Ⅲ、Ⅳ和对照Ⅰ、Ⅱ、Ⅲ、Ⅳ）,分别于1月、3月、6月检测两组大鼠肌肉组织α-KGDHC活性与蛋白表达的差异。【结果】①血糖在对照Ⅰ,Ⅱ,Ⅲ及实验Ⅰ,Ⅱ,Ⅲ之间均无显著性差异,实验组与对照组血糖相比均有显著性差异（P〈0．01）。②实验组与相应对照组α-KGDHC活性有显著性差异（P〈0．01）,三个对照组之间均无显著性差异,实验Ⅰ与实验Ⅱ之间有显著性差异（P〈0．01）,实验Ⅱ和实验Ⅲ之间无显著性差异（P〉0．05）。③随着糖尿病病程延长,肌肉组织中的α-KGDHC蛋白表达水平逐渐下降,但实验Ⅱ与实验Ⅲ之间蛋白表达未见明显差异。【结论】随着糖尿病病程的延长,糖尿病骨骼肌病变大鼠模型肌肉组织中α-KGDHC活性逐渐下降,并且与α-KGDHC蛋白水平的表达情况相一致。     

乌司他丁对肠源性脓毒症大鼠D-二聚体和蛋白C系统的影响

目的：观察乌司他丁（ulinastatin，UTI）对肠源性脓毒症大鼠D-二聚体（DD）、蛋白C（PS）系统的影响。方法：将Wistar大鼠随机分为4组，假手术组（SH组，n=36）、脓毒症组（SEP组，n=30）、UTI3h给药组（UTI3h组。n=30）和12h给药组（UTI12h组，n=18）。利用经典盲肠结扎穿孔法建立肠源性脓毒症大鼠模型，分别于正常对照组及制模后6、12、24、48及72h组心脏取血测定DD、PC和蛋白S（PS）。结果：与SH组相比，SEP组大鼠DD在造模后逐渐升高，PC、PS含量降低，差异有显著性（P〈0．05）。与SEP组相比，UTI3h,UTI12h组大鼠的DD降低，PC含量下降趋势减弱，差异有显著性（P〈0．05）。结论：UTI能有效改善脓毒症期间的抗凝抑制和继发性纤溶亢进。     

牛珀至宝丹对内毒素休克大鼠血液流变学的影响

目的：探讨牛珀至宝丹抗内毒素休克的作用机制。方法：采用静注致死量灭活大肠杆菌内毒素造成大鼠重症感染性休克模型，以牛珀至宝丹预先处理动物（实验组），并与正常组、内毒素体克组进行对照观察。结果：实验组血压下降幅度显著低于休克组；血液流变学指标中全血粘度、血浆比粘度、红细胞聚集指数、红细胞刚性指数和红细胞压积实验组较休克组均明显降低，红细胞电泳时间缩短；与正常组比较无显著性差异（Ｐ＞０．０５），但与休克组比较有显著性差异（Ｐ＜０．０５或＜０．０１）。结论：牛珀至宝丹能明显改善内毒素休克大鼠血液流变性，并对内毒素休克大鼠有一定的升压作用。     

脑缺血缺氧性模型大鼠脑红蛋白表达变化及氯化血红素的刺激效应

目的：观察脑缺血缺氧性模型大鼠脑红蛋白表达的变化及氯化血红素对其表达的影响。 方法：实验于2006-0I／03在华北煤炭医学院神经内科实验室完成。150只雄性Wistar大鼠按数字表法随机分成3组，即模型组、氯化血红素组、对照组，每组50只。①模型组：动物麻醉后切开颈前皮肤，游离双侧颈总动脉，同时阻断双侧颈总动脉血流。缺血过程中动物出现眼发白、呼吸加快、竖毛、癫痫、抽搐等并发症为模型成功。②氯化血红素组：除给予模型组同样的处理外，于造模前注射氯化血红素9次（30mol／kg．腹膜注射，3次／d）。③对照组：为假手术组，同样麻醉，剥离双侧颈总动脉，但不阻断血流。分别在血流阻断（对照组在手术后）后1，5，15，30．60min将大鼠麻醉下断头，迅速取大脑。每组每个时间点10只大鼠。各组大鼠分别行免疫组化染色和免疫蛋白印迹法，观察大鼠脑内脑红蛋白表达变化： 结果：大鼠实验过程中无死亡，150只全部进入结果分析。①模型组大鼠脑红蛋白在缺血1min表达迅速增加达高峰，5min仍维持在高水平，与对照组比较差异有显著性意义（P〈0．05）；缺血15min后迅速降低，以后又见缓慢升高，但与对照组相比差异无显著性意义（P〉0．05）。②氯化血红素干预组缺血1min和5min脑红蛋白表达快速升高，但与模型组相比差异无显著性意义（P〉0．05）；缺血15min脑红蛋白仍维持在高水平，与模型组和对照组相比差异有显著性意义（P〈0．05）；缺血30min和60min脑红蛋白已迅速降低，与对照组相比差异无显著性意义（P〉0．05）。 结论：脑缺血缺氧性模型大鼠脑红蛋白呈规律性变化；氯化血红素可以刺激脑缺血缺氧性模型大鼠脑红蛋白表达增多。     

4T超导恒定强磁场对大鼠的急性毒性作用

目的：探讨4T（40000Gs）超导恒定磁场对大鼠是否具有急性毒性作用。方法：实验于2004-03／09于中国科学院电工所超导国家重点实验室和深圳大学生命科学学院微生物基因工程深圳市重点实验室完成。选择成年健康SD大鼠130只，随机分为11组，实验大鼠分为连续处理24，48，72h3个剂量组（n=10），同时设立相应的3个对照组（n=10）。另针对磁场处理72h后，设立脱离磁场12h及1个月检测组（n=10），再设立脱离磁场作用1个月后实验大鼠一般生殖毒性实验组（n=20），并相应设立生殖毒性对照组（n=20）。采用中国科学院电工所研制的超导恒定强磁场，强度为4T。通过观察各组大鼠的一般情况、病理解剖、血液检查及生化指标检测、微核检测、染色体核型分析等，分析4T磁场对实验大鼠是否具有急性毒性作用；并且进一步观察磁场是否具有潜在或慢性生殖毒性。结果：130只大鼠全部进入结果分析，无脱失。①各组大鼠的一般情况及大体病理解剖、血常规检查指标：差异无显著性意义（F=3．14，P〉0．05）。②各组大鼠微核实验中嗜多染红细胞在红细胞中所占比率：差异无显著意义（F=2．97，P〉0．05）。③各组大鼠均无染色体丢失、臂缺损以及染色体带型变化。④大鼠的生殖毒性实验结果：生殖毒性组与生殖毒性对照组大鼠（雄性）精子畸形数量及形态比较，差异无显著性意义（F=2．62，P〉0．05）；生殖毒性组与生殖毒性对照组大鼠（雌性）受孕率、着床数、活胎数、死胎数、吸收胎数变化及胎仔雌雄比例、体质量、体长、尾长、囱门宽度比较，差异无显著性意义（F=2．15，P〉0．05）。结论：4T超导恒定磁场对实验大鼠无急性毒性作用，目前临床所使用的磁疗强度远低于此，对于生物体是安全的。     

AVE-764B尿液有形成分分析仪的性能评价及临床应用价值

目的对AVE-764B尿液有形成分分析仪的性能进行评价,评估其是否能用于尿液有形成分检测。方法对仪器的精密度、分析测量范围、携带污染、阳性检出符合率、干扰试验及315份临床尿液标本红细胞、白细胞参数与人工镜检法结果比对,对五大性能指标进行验证和评价。结果仪器的批内精密度高、中、低值分别为6.93%、8.95%、12.33%;批间精密度高、中、低值分别为7.64%、9.72%、12.48%;红细胞在(0～18 812)个/微升线性范围相关系数(r)为0.987,白细胞在(0～2 020)个/微升线性范围r为0.972;红细胞、白细胞阳性检出符合率分别为100%、90%;红细胞、白细胞携带污染率分别为0.06%、0.01%;干扰试验:草酸钙结晶不干扰仪器红细胞的检测结果;315份临床尿液标本采用Kappa一致性检验进行仪器与人工镜检法结果比较,红细胞:U=8.95,P<0.05;白细胞:U=41.49,P<0.05,显示两种检测方法结果一致。但在本研究的315例尿液标本中,AVE-764B尿液有形成分分析仪对环形红细胞容易漏检,经厂家技术人员多次调整参数后,又对54例镜下血尿标本用上述两种方法进行检测比较,红细胞:U=5.09,P<0.05,两种检测方法具有一定的一致性。结论 AVE-764B尿液有形成分分析仪可用于尿液有形成分检测,但必须把仪器的参数调节在最佳状态,使用前用含环形红细胞的血尿标本进行验证,确保仪器能拍摄到环形红细胞,以免漏检环形红细胞,并定期验证,以避免计数池变脏而漏检。     

Use of a flow-cell system to investigate virucidal dimethylmethylene blue phototreatment in two RBC additive solutions

BACKGROUND: Limited photoinactivation kinetics, use of low-volume 30 percent Hct RBCs, and hemolysis have restricted the practicality of the use of dimethylmethylene blue (DMMB) and light for RBC decontamination. A flow-cell system was developed to rapidly treat larger volumes of oxygenated 45 percent Hct RBCs with high-intensity red light. MATERIALS AND METHODS: CPD-whole blood was WBC reduced, RBCs were diluted in additive solutions (either Adsol or Erythrosol), and suspensions were subsequently oxygenated by gas overlay. Intracellular or extracellular VSV and DMMB were sequentially added. VSV-infected RBC suspensions (45% Hct) were passed through 1-mm-thick flow cells and illuminated. Samples were titered for VSV, stored for up to 42 days, and assayed for Hb, supernatant potassium, ATP, and MCV. RESULTS: The use of oxygenated RBCs resulted in rapid and reproducible photoinactivaton of > or = 6.6 log extracellular and approximately 4.0 log intracellular VSV independent of additive solution. Phototreated Adsol RBCs exhibited more than 10 times greater hemolysis and 30 percent greater MCV during storage than identically treated Erythrosol RBCs. Phototreatment caused RBC potassium leakage from RBCs in both additive solutions. ATP levels were better preserved in Erythrosol than Adsol RBCs. CONCLUSION: A rapid, reproducible, and robust method for photoinactivating model virus in RBC suspensions was developed. Despite improved hemolysis and ATP levels in Erythrosol-phototreated RBCs, storage properties were not maintained for 42 days.     

血栓性血小板减少性紫癜2例报道

目的总结兰州大学第二医院2例血栓性血小板减少性紫癜(TTP)的所有实验检测资料。方法回顾性分析与TTP诊断特征密切相关的实验室检测:血红蛋白(Hb)、血小板(PLT)、网织红细胞(RET)、裂片红细胞(FRC)、尿蛋白(PRO)、尿潜血(BLD)、乳酸脱氢酶(LDH)、总胆红素(TBil)、直接胆红素(DBil)、间接胆红素(IBil)、尿素氮(BUN),肌酐(Cr)、D二聚体(D-D)及骨髓检测,并归纳分析。结果血浆、PLT、静丙输注无法改善PLT的严重减少;输注红细胞(RBC),Hb进行性下降;尿液PRO及BLD定性持续阳性;溶血检测中的RET、TBil、IBil及LDH持续增高;凝血检测中D-D持续不降;骨髓分析提示2例患者分别是增生性贫血与溶血性贫血并考虑Evans综合症。结论对于TTP浅显认识导致临床诊治延误。回顾性总结TTP实验室检测及临床特征,发现有经典"五联征",包括血小板减少症、微血管病性溶血性贫血、神经症状、肾脏损害及发热。     

Use of a flexible thiopyrylium photosensitizer and competitive inhibitor for pathogen reduction of viruses and bacteria with retention of red cell storage properties

BACKGROUND: Progress in developing photochemical methods for pathogen reduction of red blood cells (RBCs) has been hampered by hemolysis. A flexible, nucleic acid-intercalating thiopyrylium (TP) dye that is only photochemically active in the bound state and a competitive inhibitor of RBC membrane binding, dipyridamole (DP), was used to reduce photoinduced hemolysis stemming from free- and membrane-bound dye. STUDY DESIGN AND METHODS: Oxygenated leukodepleted 20% hct RBC suspensions were deliberately inoculated with virus or bacteria, incubated with 200 micromol per L DP and less than or equal to 100 micromol per L TP, illuminated with 1.1 J/cm(2) of red light, and titered. RBC suspensions containing 200 micromol per L DP and 160 micromol per L TP were identically phototreated, concentrated to 45% hct, and assayed for RBC storage properties. RESULTS: In RBC suspensions containing DP, TP photoinactivated vesicular stomatitis virus, pseudorabies virus, duck hepatitis B virus, bovine virus diarrhea virus, extracellular human immunodeficiency virus (HIV) to the limit of detection and 6.2 log intracellular HIV. More than 5 log inactivation of 6 bacterial species was demonstrated. DP prevented approximately 30% of TP binding to RBCs. Phototreated RBCs that were subsequently stored for 42 days exhibited acceptable levels of hemolysis, morphology scores, extracellular pH, ATP, glucose utilization rates, and lactate production. Treated samples exhibited substantially increased potassium efflux compared to controls. CONCLUSION: Use of TP photosensitizer and DP enables significant levels of pathogen reduction while retaining most, but not all RBC properties during 42 day storage.     

ABO血型主要不合供者外周血干细胞单采中COM.TEC血细胞分离机的应用

背景：异基因外周血造血干细胞移植大约1/3是在ABO血型不合的供受者之间进行的。目的：探讨ABO血型主要不合供者外周血造血干细胞在不去除红细胞情况下应用COM.TEC血细胞分离机直接回输给受者的可行性。方法：27例HLA全相合同胞供者皮下注射粒细胞集落刺激因子5μg/（kg.d）,连续5d干细胞动员,于第5天开始应用COM.TEC血细胞分离机进行外周血造血干细胞单采,每次总处理血量为3.0～3.5个供者全身血量。ABO相合供者按常规方法进行单采,主要不合供者采取动态调整白膜收集量和保持稳定充足的血流量以减少红细胞污染。结果与结论：27例供者中ABO主要不合供者11例,相合供者16例。主要不合组产品体积及红细胞内含量均显著低于相合组（P〈0.05）,回输给受者均未出现急性溶血反应;2组单个核细胞及CD34＋细胞计数差异无显著性意义（P〉0.05）,受者均获造血重建,中性粒细胞及血小板重建时间相似（P〉0.05）。提示应用COM.TEC血细胞分离机进行ABO主要不合供者外周血造血干细胞单采,可以保持稳定充足的血流量情况下通过动态调整白膜收集量参数,在不影响采集产品产量和质量的同时显著减少红细胞的污染,预防回输后受者的急性溶血反应。     

A comparison of hemolysis and red cell mechanical fragility in blood collected with different cell salvage suction devices

BACKGROUND: Suction pressure is one variable that can affect the efficiency of red blood cell (RBC) recovery for intraoperative autotransfusion. This study compared a constant-suction-pressure system with a device that is expected to minimize hemolysis by automatically varying the suction pressure. STUDY DESIGN AND METHODS: Twenty-two 50-mL reconstituted whole-blood samples were collected from a flat surface with either a constant-pressure suction device (BRAT 2 autologous blood recovery system, COBE Cardiovascular) set to a pressure of -200 mmHg or a variable-pressure suction device (SmartSuction Harmony surgical suction pump, Haemonetics, Inc.). Time of blood collection was recorded and plasma free hemoglobin (PFHb) was spectroscopically measured in the aspirated blood. Postcollection blood was subjected to RBC mechanical fragility test to characterize potential sublethal blood trauma. Relative hemolysis and the mean fragility index (MFI) were calculated. RESULTS: Hemolysis was significantly reduced with the variable-pressure suction compared to the constant-pressure suction (2.17% vs. 3.20%, respectively; p < 0.001). There was no significant difference between both suction devices in either the MFI of RBCs (0.632 vs. 0.673, respectively; p > 0.05) or the collection time at -200 mmHg (54.2 seconds vs. 52.8 seconds, respectively; p > 0.05). CONCLUSIONS: Although the variable-pressure device produced a significant reduction in hemolysis during one-pass blood collection, the clinical significance of this reduction is not clear. In relative terms, the variable-pressure device would recover an extra 10 mL of RBCs for every liter of salvaged RBCs, which is negligible compared to the blood loss in major surgery.     

Critical role of phosphatidylserine in hemolysis due to red blood cell enzyme/membrane defects

The life span of human red blood cells (RBCs) is approximately for 120 days, and finally destroyed in reticuloendothelial systems. In pathological conditions, RBCs fall into premature death, i.e. hemolysis. Recent studies have unveiled that abnormal RBCs affected by either membrane disorders or enzyme deficiency as well as senescent RBCs shared the common feature, i.e. exposure of phosphatidylserine (PS) in the outer leaflet of the RBC membrane. Increased intracellular oxidative stress or enhanced calcium permeability impairs maintenance of phospholipid asymmetry, resulting in PS externalization. PS and other eat-me signals may contribute to recognition of abnormal RBC by macrophage. In this review, recent understanding of mechanisms underlying hemolysis are discussed.     

Differences in Rat and Human Erythrocytes Following Blood Component Manufacturing: The Effect of Additive Solutions

Background

Small animal models have been previously used in transfusion medicine studies to evaluate the safety of blood transfusion products. Although there are multiple studies on the effects of blood banking practices on human red blood cells (RBCs), little is known about the effect of blood component manufacturing on the quality of rat RBCs.

Methods

Blood from Sprague-Dawley rats and human volunteers (n = 6) was collected in CPD anticoagulant, resuspended in SAGM or AS3, and leukoreduced. In vitro quality was analyzed, including deformability, aggregation, microvesiculation, phosphatidylserine (PS) expression, percent hemolysis, ATP, 2,3-DPG, osmotic fragility, and potassium concentrations.

Results

Compared to human RBCs, rat RBCs had decreased deformability, membrane rigidity, aggregability, and microvesiculation after component manufacturing process. Rat RBCs in SAGM showed higher hemolysis compared to human RBCs in SAGM (rat 4.70 ± 0.83% vs. human 0.34 ± 0.07%; p = 0.002). Rat RBCs in AS3 had greater deformability and rigidity than in SAGM. The number of microparticles/µl and the percentage PS expression were lower in rat RBCs in AS3 than in rat RBCs in SAGM. Hemolysis was also significantly lower in AS3 compared to SAGM (2.21 ± 0.68% vs. 0.87 ± 0.39%; p = 0.028).

Conclusion

Rat RBCs significantly differ from human RBCs in metabolic and membrane-related aspects. SAGM, which is commonly used for human RBC banking, causes high hemolysis and is not compatible with rat RBCs.Key Words: Red blood cells, Additive solutions, Blood manufacturing, Blood banking     

Exploratory in vitro study of red blood cell storage containers formulated with an alternative plasticizer

BACKGROUND: The plasticizer di‐2‐ethylhexyl phthalate (DEHP) is a common component in medical plastics. There is motivation to replace this component; however, DEHP is necessary to prevent excessive hemolysis in stored red blood cells (RBCs). Our objective is to evaluate a candidate replacement plasticizer (Hexamoll, di‐isononyl cyclohexane‐1,2‐dicarboxylic acid [DINCH], BASF Corp.) compared to DEHP in an in vitro feasibility study. We hypothesize that the candidate will provide at least equivalent protection against hemolysis for RBCs stored for 42 days and periodic mixing of RBCs will add additional protection against hemolysis. STUDY DESIGN AND METHODS: Whole blood was collected into citrate‐phosphate‐dextrose; combined into pools of 2 ABO identical whole blood units; and divided, leukoreduced, centrifuged, and separated into plasma and RBCs. Additive solution was added, and the RBCs were stored for 42 days at 1 to 6°C. In three parts of this study, split pools were paired as DINCH‐polyvinyl chloride (PVC) with weekly mixing versus DINCH‐PVC with no mixing, DINCH‐PVC mixed versus DEHP‐PVC no mix, and DINCH‐PVC versus DEHP‐PVC with neither mixed. A standard panel of in vitro RBC characteristics was determined on Days 0 and 42. RESULTS: Mixing DINCH‐PVC weekly improved Day 42 hemolysis (0.36 ± 0.07% vs.0.56 ± 0.15%, p = 0.002), and mixed DINCH‐PVC bags were noninferior to unmixed DEHP‐PVC bags (p ≤ 0.05). DINCH‐PVC bags stored without weekly mixing were inferior to unmixed DEHP‐PVC bags for hemolysis on Day 42, although no individual bag exceeded 0.8% hemolysis. CONCLUSION: Periodic mixing of RBCs stored in DINCH‐PVC provides additional protection against hemolysis. Unmixed DINCH‐PVC bags were inferior to DEHP‐PVC bags for prevention of hemolysis, but remain a candidate for replacement DEHP in RBC storage bags.     

Thiazole orange, a DNA-binding photosensitizer with flexible structure, can inactivate pathogens in red blood cell suspensions while maintaining red cell storage properties

BACKGROUND: Development of a robust pathogen reduction system for red cells (RBCs) utilizing photosensitive dyes has been constrained by hemolysis, usually mediated by reactive oxygen species emanating from dye free in solution as well as dye bound to the RBC membrane. The RBC binding properties of thiazole orange (TO), a flexible nucleic acid intercalating cyanine dye that predominantly acts as a photosensitizer only when bound, were assessed along with its virucidal, bactericidal, and light-induced hemolytic activities. STUDY DESIGN AND METHODS: Leukodepleted 20% hematocrit RBCs suspended in Erythrosol (RAS-2) were oxygenated, inoculated with test organisms, incubated with TO, and illuminated. Control and treated samples were analyzed by appropriate assay. Identically prepared, but uncontaminated samples were phototreated, concentrated to 45% hematocrit, and assayed for potassium leakage, hemolysis, and ATP during storage. RESULTS: Approximately 21 percent TO bound to RBCs. Phototreatment inactivated from 5.4 to 7.1 log(10) of 5 tested viruses and from 2.3 to greater than 7.0 log(10) of 8 tested bacteria. Phototreated RBCs exhibited only slightly increased hemolysis, moderately elevated potassium efflux, and similar levels of ATP compared to controls. CONCLUSION: TO can photoinactivate several model viruses and pathogens in RBCs under conditions that produce limited hemolysis without the addition of quenchers or competitive inhibitors.