不同洗涤液制备冰冻解冻去甘油红细胞的检测结果分析

目的通过用两种洗涤溶液制备冰冻解冻去甘油红细胞结果进行质量分析,来比较应用不同洗涤溶液,一类是9%氯化钠、羟乙基淀粉130氯化钠注射液、0.9%氯化钠;一类是9%氯化钠、0.9%氯化钠分别进行解冻红细胞洗涤,选择合适制备冰冻解冻去甘油红细胞的洗涤方法。方法随机抽取2010年采血6d之内新鲜全血32份,通过手工制备冰冻红细胞：6d之内（2～6℃保存）新鲜全血经过25min,2300转,离心后去除血浆,将血液倒入三珠空袋内,用无菌接驳机连接输血器,应用甘油160ml,25min加入振荡器60次/min,室温沉淀30min后放入-80℃冰箱速冻,1个月后进行解冻。随机分为A、B两组,每组16袋,应用ACP215血液处理仪进行解冻红细胞。A组洗涤液包括9%氯化钠、羟乙基淀粉130氯化钠、0.9%氯化钠这三种溶液;B组洗涤液包括9%氯化钠、0.9%氯化钠这两种溶液。用两种不同洗涤溶液进行冰冻解冻去甘油红细胞洗涤,通过检测项目为红细胞回收率%,白细胞残留率%,血小板残留率%,甘油含量,血浆游离血红蛋白含量,体外溶血等指标进行检测分析。结果 A组解冻红细胞：红细胞回收率81.0%±2.6%,白细胞残留率0.67%±0.14%,血小板残留率0.00%,甘油含量4.8±0.43g/L,血浆游离血红蛋白含量0.62±0.11g/L,体外溶血13%±5%。B组解冻红细胞：红细胞回收率82.0%±2.3%,白细胞残留率0.61%±0.26%,血小板残留率0.42%±0.1%,甘油含量（5.1±0.13）g/L,血浆游离血红蛋白含量（0.53±0.12）g/L,体外溶血12%±4%。两组结果均符合国家标准,且无统计学意义。结论通过手工制备冰冻红细胞后,应用ACP215血液处理仪制备冰冻解冻去甘油红细胞分别使用A、B两种洗涤液,根据统计学分析洗涤效果无统计学意义,检测结果均达到国家标准,可以确定在洗涤中不加入羟乙基淀粉130氯化钠注射液,检测结果也能达到国家标准。     

关于冰冻红细胞制备、洗涤过程的探讨

目的 采用两种方法洗涤冰冻解冻去甘油红细胞,比较它们对产品质量的影响,为冰冻解冻去甘油红细胞的制备提供质量依据.方法 对（-70±5）℃保存30 d的10袋冰冻解冻去甘油红细胞,解冻后采用两种洗涤方法处理,计算出洗涤前后各袋血血红蛋白含量等指标.结果 方法一用ACP215型血液处理机渗透压变化率设为350,洗涤3次方法洗涤的红细胞的血红蛋白含量高于方法二ACP215型血液处理机渗透压变化率设为500,洗涤5次的红细胞,但上清游离血红蛋白含量方法一高于方法二,其他指标接近.结论 方法二洗涤效果较好.     

Rh阴性血液深低温保存的质量控制

目的探讨保证深低温保存Rh阴性红细胞(RBC)解冻后的质量检测方法。方法用ACD抗凝,4℃保存6d内Rh阴性RBC悬液或全血离心除去添加剂或血浆的浓缩RBC,将RBC经40％甘油处理后,置-80℃冰冻保存。临床需要时37℃水浴解冻复苏RBC,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悬浮RBC。结果26袋冰冻解冻RBC去甘油后RBC回收率为(81．17±18．5)％,游离血红蛋白为(0．63±0．16)g/L,甘油残留量平均为5．3g／L,体外溶血实验血红蛋白增加率为25．19％。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。     

全自动细胞处理系统制备与手工制备解冻红细胞质量指标比较

目的观察用全自动细胞处理系统(ACP215)与手工方法制备冰冻解冻红细胞后的质量指标,比较2种方法制备的冰冻解冻红细胞的差异。方法随机取需冰冻保存的红细胞制剂分2组:A组25例用手工方法直接滴入红细胞低温保护剂;B组25例用全自动细胞处理系统全自动加入红细胞低温保护剂,然后分别放置在-80℃低温冰箱冰冻保存4周以上,在临床需要使用时取出融化;分别使用手工方法制备和全自动细胞处理系统制备解冻红细胞。并分别对冰冻解冻红细胞的红细胞回收率、游离血红蛋白、甘油残留量的指标进行检测。结果使用全自动细胞处理系统(ACP215)保存、洗涤的冰冻解冻红细胞比手工法制备的冰冻解冻红细胞其红细胞回收率有所升高,甘油残留量有所下降,而上清液中游离血红蛋白浓度没有明显变化。结论使用全自动细胞处理系统(ACP215)冰冻保存、解冻去甘油红细胞操作方便、洗涤时间短,产品质量要优于手工方法。     

红细胞冰冻保存的研究和应用

用高浓度甘油将人红细胞（RBC）甘油化，置-80℃冰冻保存。用前于37℃水浴快速解冻，并用盐液-SAGM液（葡萄糖、腺嘌呤、甘露醇和氯化钠）洗涤红细胞。测定2．3-DPG、甘油残余量、溶血率和回收率，以及RBC脆性试验等。考查RBC保存质量均符合国家标准并应用于临床。     

制备洗涤红细胞方法关键控制点改进体会

洗涤红细胞是指采用物理方法在无菌条件下,将保存期内的全血、浓缩红细胞、悬浮红细胞血液制剂用大量静脉注射0.9%氯化钠溶液洗涤,去除大部分非红细胞部分,并将红细胞悬浮在0.9%氯化钠溶液中,所制成的红细胞成分血,其特点：洗涤红细胞是在少白细胞红细胞的基础上用无菌0.9%氯化钠溶液反复洗涤3遍以上制备而成的,去除了80%以上的白细胞和98%的血浆蛋白,也去除了的大量的细胞碎屑、代谢产物、抗凝剂、钾、氨和微聚物,同时也损失了约20%的红细胞。     

微波水热法解冻冰冻红细胞在骨科患者中的临床应用

目的:探讨微波水热法解冻快速冰冻红细胞,用于骨科患者的安全性和有效性.方法:用微波水热法解冻以甘油为保护剂-80℃保存的红细胞,用不同浓度的洗涤液洗涤去除甘油,以红细胞回收率、游离血红蛋白含量、残余甘油含量、体外溶血试验和无菌试验为质量检测控制项目.输血过程中观察患者有无发热、过敏、溶血及血压降低等不良反应并纪录输血速度;患者输血后,检测24 h、72 h血常规,生化及血气分析.结果:冰冻红细胞质量达到<全血及成分质量要求>(GB18469-2001)的要求.患者输血后未发现不良事件,各项指标72 h内存在一定的变化,但均在正常范围内,24 h、72 h各项指标与输血前相比,RBC、HGB未明显下降,肝、肾功能未受明显损害,血K+、血气各项指标无明显变化.结论:微波水热法能快速解冻冰冻红细胞,质量符合标准,应用于骨科患者安全、有效,为解决血液长期储存、保障供血和稀有血型患者输血及自身输血提供了新的方法和手段.     

强冷凝集对血细胞干扰1例处理方法探讨及文献复习

探讨强冷凝集素补体C3对血细胞红系参数的影响及处理方法,以便获得可靠的处理结果。筛选1例骨髓移植术4个月后地贫患者,针对该患者补体C3强阳性样本,分别在室温（25℃）、37℃水浴30 min、37℃水浴60 min、41℃水浴1 min、41℃水浴30 min、45℃水浴1 min、45℃水浴30 min、45℃生理盐水洗涤红细胞3次等方法依次处理,然后采用迈瑞BC6800血细胞分析仪进行常规检测,同时进行手工涂片染色镜检。红细胞（RBC）、血红蛋白（HGB）、血细胞比容（HCT）、平均红细胞体积（MCV）、平均红细胞血红蛋白含量（MCH）、平均红细胞血红蛋白浓度（MCHC）均有不同程度改变。在日常实验操作中,如遇到强冷凝集素补体C3存在引起血细胞分析困难时,采用45℃水浴1 min或45℃生理盐水洗涤3次,然后进行血常规检测,均可获得符合临床的结果。实际操作中采用45℃生理盐水洗涤,操作比较繁琐,而且离心温度较难掌控,而45℃水浴1 min不仅操作简单,结果也较准确,保证了血常规检测结果的准确性。     

冰冻红细胞去甘油方法学评价

目的通过比较多功能细胞淘洗机和手工制备冰冻解冻去甘油红细胞去甘油化的效果评价,找到一种速度快、效率高的洗涤方法。方法作者收集利用多功能细胞淘洗机和手工冰冻解冻去甘油红细胞洗涤各48份,按《全血成分血质量要求》标准检测冰冻解冻去甘油红细胞回收率、上清游离血红蛋白、体外溶血试验,上清甘油含量。结果细胞淘洗机系统去甘油洗涤快速、实用,克服了手工洗涤操作步骤多、时间长、效率低,人为因素影响大等问题。两种方法制备冰冻解冻去甘油红细胞均符合质量标准,多功能细胞淘洗机所需时间短,工作效率高。结论通过质控检测比较,功能细胞淘洗机快速、实用,适用于RhD冰冻红细胞的去甘油化,适合血站使用。     

冰冻解冻红细胞的制备、保存及临床应用

目的通过探讨10份-65℃以下保存冰冻红细胞制备解冻红细胞的方法,并对其影响质量的相关因素进行分析。方法使用ACP215血液处理机制备冰冻解冻红细胞。结果应用该方法制备的解冻红细胞经检测,红细胞回收率、残留白细胞、残留血小板、残留甘油含量、上清游离血红蛋白量及体外溶血试验等质量因素差异均无显著性且均达到了国家标准。结论该方法适用于临床上稀有血型患者的输血需求。操作简便,高效,冰冻红细胞保存时间长,质量稳定,有效提高了临床用血的安全性。     

安阳地区血液制剂容量控制中标示量范围的确定

目的探讨安阳地区血液制剂容量控制中标示量的范围，为临床输血工作提供可靠的数据支持。方法根据《全血及成分血质量要求GB18469—2001》中容量标准要求结合安阳地区血液制剂制备实际情况，确定出符合安阳地区血液制剂容量标准的标示量范围，并将确定出的标示量范围与实际制备的血液制剂容量进行比对。结果安阳地区来源于400ml全血的去白全血容量标示范围确定为（460±46）ml；来源于400ml全血的2U去白悬浮红细胞容量标示范围确定为（300±30）ml；来源于200ml全血的1U去白悬浮红细胞容量标示范围确定为（150±15）ml。安阳地区实际制备的血液制剂容量85％以上都在确定的标示量范围内。结论安阳地区血液制剂容量符合国家标准要求，建议在血液制剂标签上注明标示量范围，血液制剂标示量范围的准确标定具有重要的临床意义。     

The erythrocyte transport and transfer of methylmercury to the tissues of the rainbow trout (Salmo gairdneri).

Methylmercury (MeHg) was found to be taken up rapidly and almost completely by trout red blood cells (RBC) both in vitro and in vivo. The binding of MeHg within the RBC was freely reversible both in vitro, as shown by the efflux of MeHg from RBCs suspended in protein solutions, and in vivo following intracardial (i.c.) injection of RBC-bound MeHg. Hemoglobin (Hb) appeared to be the main MeHg transport protein in trout blood since it bound 90% of whole blood Hg following an intragastric dose of Me203HgCl. MeHg, injected i.c. as MeHgS-cysteine, was found to be present in blood bound almost completely to hemoglobin 10 days post-injection. This suggests an ability of hemoglobin to compete for and bind MeHg bound to other sulfhydryl (-SH) compounds. The number of reactive -SH groups per molecule of trout Hb was determined to be 4 by amperometric titration with MeHgCl. The concentration of Hb reactive -SH groups in the trout RBC was calculated to be at least 20 mM. This accounts for the high affinity of the RBC for MeHg.     

The effect of pentoxifylline on filterability of normal red blood cells and their adhesiveness to cultured endothelial cells

Summary The effects of pentoxifylline on filterability of normal red blood cells (RBCs) and their adhesiveness to cultured endothelial cells were investigated.1. In a balanced randomized and double blind trial, six healthy volunteers received 400 mg pentoxifylline or matching placebo 2 h before blood samples were taken. Filterability of RBCs of the subject while on pentoxifylline was significantly increased at 25 °C and 18 °C.2. Lowering of the filteration temperature to 18 °C significantly decreased filterability of RBCs.3. In vitro studies showed that 12 µg/ml pentoxifylline significantly increased RBC filterability and also partially prevented the effect of decreasing temperature on RBC filterability.4. 12 µg/ml pentoxifylline significantly decreased the adherence of normal RBCs to cultured endothelial cells.Our results suggest that in addition to increasing filterability of RBCs, pentoxifylline also decreased the adherence of RBCs to endothelial cells and this may contribute to its therapeutic effect.     

离子对高效液相色谱法测定人体红细胞内核苷二磷酸激酶活性

目的:建立测定人体红细胞(RBC)内核苷二磷酸激酶(NDPK)活性的反相离子对高效液相色谱法。方法:稀释RBC中加入反应底物(dADP和dGTP)及其他反应混合液后,37℃孵育5 min,100℃灭活10 min,离心取上清液进样分析。以反应产物dATP定量分析NDPK活性。色谱柱为SymmetryShield TMRP18(150 mm×3.9 mm,5μm),检测波长为260 nm,柱温为25℃;采用梯度洗脱,流动相A为20 mmol.L-1 KH2PO4-K2HPO4缓冲液、加入离子对试剂5 mmol.L-1四丁基硫酸氢铵(TBAHS)溶液,流动相B为乙腈。结果:NDPK催化反应dADP+dGTPdATP+dGDP反应特异性良好,RBC内dATP的线性范围为4.33～43.34μmol.L-1(r2=1.000),定量下限为4.33μmol.L-1。日内及日间RSD分别小于5.53%和6.69%,准确度为98.4%～104.2%,平均绝对回收率大于83%;dATP样品在室温或4℃、-80℃保存30 d及反复冻融3次稳定性良好。结论:该法操作简便快速、灵敏、专属性强,可应用于人体RBC内NDPK活性测定,为研究NDPK活性与嘌呤类药物反应相关性提供基础。     

冰冻亚型红细胞信息库的建立及应用

目的探讨冰冻亚型红细胞信息库的建立及应用,以利于冰冻亚型红细胞的合理选择和应用。方法以甘油作为防冻剂,将ABO亚型红细胞进行甘油化和适当分装后,于-80℃低温冰箱冰冻保存;同时详细记录每份红细胞的来源、亚型种类、弱抗原与抗血清的反应强度、红细胞容量、预期用途、分装容器、分装总份数、每份分装量、血浆是否含ABO不规则抗体、是否保存对应血浆以及献血码或姓名、性别、年龄、民族、身份证号码、通讯地址、联系电话等资料,建立冰冻亚型红细胞信息库;根据不同工作需要,从红细胞信息库中选择合适的亚型红细胞,经融化洗涤,加入合适的保存介质和抑菌剂后使用。结果从冰冻亚型红细胞信息库中选择合适的亚型标本,在血站和医院供血库ABO血型鉴定相关工作中应用后,无偿献血者和医院患者的ABO亚型检出率显著提高,进一步保证了临床输血的安全性和有效性。结论 ABO亚型红细胞的冰冻保存及其信息库的建立,能使ABO亚型红细胞得到充分有效的利用,具有良好的经济效益和社会效益,可在采供血机构和医院供血库进行推广应用。     

阿米福汀对骨髓增生异常综合征患者红细胞寿命的影响

目的研究骨髓增生异常综合征(myelodysplastic syndrome,MDS)患者的红细胞寿命以及阿米福汀对红细胞寿命的影响。方法回顾性分析浙江省中医院血液科2017年1月-2018年7月期间81例MDS患者的临床资料,分为阿米福汀治疗组(49例)和支持治疗组(32例)。以同期9名正常志愿者为正常对照,采用内源性一氧化碳呼气试验法测定患者及志愿者的红细胞寿命,分析MDS患者红细胞寿命及阿米福汀对红细胞寿命的影响。结果 81例初发MDS患者(未经输血支持)平均红细胞寿命为(33.41±10.96)d,与正常对照红细胞寿命(121.11±32.59)d相比明显缩短(P<0.01)。阿米福汀治疗组:部分缓解1例,血液学改善13例,疾病稳定12例,总有效率53.1%。治疗后MDS患者的红细胞寿命(42.24±12.99)d,与治疗前相比明显延长(P<0.01)。支持治疗组:血液学改善2例,疾病稳定9例,总有效率34.4%,红细胞寿命(34.53±7.50)d,与治疗前相比未明显改善。红细胞寿命与疗效关系分析表明,血液学改善患者的红细胞寿命(45.92±9.24)d,较正常对照组红细胞寿命短(P<0.01),与治疗前比较明显延长(P<0.01)。其余患者红细胞寿命治疗前后无统计学意义。结论 MDS患者红细胞寿命缩短,阿米福汀治疗后可提高患者红细胞寿命。     

外周血涂片检出幼稚红细胞临床意义探讨

目的通过对外周血涂片幼稚红细胞检出率分析，探讨血液肿瘤性疾病与血液非肿瘤性疾病外周血涂片出现幼稚红细胞的临床意义。方法对2001年1月～2007年10月我院992例骨髓细胞学检查患者同时做外周血涂片检查。血涂片计数100个白细胞并分类，幼稚红细胞检出率以100个白细胞同时发现幼稚红细胞数（即幼红细胞数／100个WBC）表示。结果在992例骨髓和外周血同步检查中，外周血涂片检出幼稚红细胞122例。血液肿瘤性疾病外周血涂片幼红细胞检出率为32．3％，血液非肿瘤性疾病外周血涂片幼红细胞检出率为9％，两者外周血涂片幼红细胞检出率比较差异有显著性（P〈0.05）。结论外周血涂片检出幼稚红细胞对血液肿瘤性疾病与血液非肿瘤性疾病的诊断与鉴别诊断有一定的临床意义，血液中出现不明原因的幼红、幼粒细胞应引起重视，严密随访。     

In vitro and ex vivo effects of indobufen on red blood cell deformability

Summary We have studied the effect of indobufen, a cyclo-oxygenase blocking agent which has proved useful in patients with obstructive vascular disease, on red blood cell (RBC) filterability in vitro and in a pilot study ex vivo.The addition of indobufen in vitro to blood samples from 10 healthy volunteers did not significantly modify RBC deformability.We evaluated the ex vivo effect of indobufen (200 mg bd) in 14 patients with obstructive vascular disease. A significant improvement in RBC deformability was noted on the 5th, 14th, and 28th days of treatment, 2 h after the morning dose. Acetylsalicylic acid given to 6 similar patients had no effect suggesting that the positive haemorheological effect of indobufen is probably not linked to its cyclooxygenase blocking effect.     

An Isolated In-Situ Rat Head Perfusion Model for Pharmacokinetic Studies

Purpose. To develop a viable, single pass rat head perfusion modeluseful for pharmacokinetic studies. Methods. A viable rat head preparation, perfused with MOPS-bufferedRinger's solution, was developed. Radiolabelled markers (red bloodcells, water and sucrose) were injected in a bolus into the internalcarotid artery and collected from the posterior facial vein over 28minutes. The double inverse Gaussian function was used to estimatethe statistical moments of the markers. Results. The viability of the perfusion was up to one hour, with optimalperfusate being 2% bovine serum albumin at 37°C, pH 7.4. Thedistribution volumes for red blood cells, sucrose and water (from all studies,n = 18) were 1.0 ± 0.3ml, 6.4 ± 4.2ml and 18.3 ± 11.9ml, respectively.A high normalised variance for red blood cells (3.1 ± 2.0) suggestsa marked vascular heterogeneity. A higher normalised variance forwater (6.4 ± 3.3) is consistent with additional diffusive/permeabilitylimitations. Conclusions. Analysis of the physiological parameters derived fromthe moments suggested that the kinetics of the markers were consistentwith distribution throughout the head (weight 25g) rather than justthe brain (weight 2g). This model should assist in studying solutepharmacokinetics in the head.     

Human thiopurine methyltransferase activity varies with red blood cell age

AIMS: Inherited differences in thiopurine methyltransferase (TPMT) activity are an important factor in the wide interindividual variations observed in the clinical response to thiopurine chemotherapy. The aim of this study was to establish a population range for red blood cell (RBC) TPMT activity in children with acute lymphoblastic leukaemia (ALL) at disease diagnosis. An additional aim was to investigate factors that can influence TPMT activity within the RBC. METHODS: Blood samples were collected from children with ALL at disease diagnosis, prior to any blood transfusions, as part of the nationwide UK MRC ALL97 therapeutic trial. RBC TPMT activity was measured by h.p.l.c. RBCs were age-fractionated on Percoll density gradients. RESULTS: Pretreatment blood samples were received from 570 children within 3 days of venepuncture. TPMT activities at disease diagnosis ranged from 1.6 to 23.6 units/ml RBCs (median 7.9) compared with 0.654-18.8 units (median 12.9), in 111 healthy control children (median difference 4.5 units, 95% CI 3.9, 5.1 units, P < 0.001). A TPMT quality control sample, aliquots of which were assayed in 60 analytical runs over a 12 month period, contained a median of 11.98 units with a CV of 11.6%. Seven children had their RBCs age-fractionated on density gradients. TPMT activities in the top gradient (young cells) ranged from 4.2 to 14.1 units (median 7.5) and in the bottom gradient (old cells) 1.5-12.6 units (median 4.7 units), median difference 2.3 units, 95% CI 0.7, 4.1, P = 0.035. CONCLUSIONS: Circulating RBCs do not constitute a homogeneous population. They have a life span of around 120 days and during that time undergo a progressive ageing process. The anaemia of ALL is due to deficient RBC production. The results of this study indicate that RBC TPMT activities are significantly lower in children with ALL at disease diagnosis. This may be due, at least in part, to a relative excess of older RBCs.