单采血小板中红细胞混入量超标原因调查分析

目的:探讨单采血小板中红细胞混入量超标的原因。方法:根据所采集的单采血小板是否红细胞混入量超标(有肉眼可见红细胞),分设正常对照组与超标组,正常对照组为红细胞混入量符合标准的献血者80例;超标组为红细胞混入量超标的献血者23例。分别对2组献血者样本进行全血常规及血红蛋白项目检测。结果:超标组中献血者的Hb、Hct、MCV、MCH及血浆总蛋白结果均低于正常对照组献血者,差异有统计学意义(P0.01);而2组献血者RBC计数、MCHC比较差异无统计学意义(P0.05)。结论:单采血小板红细胞混入量超标与外周血的Hb、Hct、MCV、MCH以及血浆总蛋白等参数的水平降低有关;与献血者多次献血或混合献血造成红细胞未完全成熟有关。     

Accuracy of the automated cell counters for management of spontaneous bacterial peritonitis

AIM： To evaluate the accuracy of automated blood cell counters for ascitic polymorphonuclear （PMN） determination for： （1） diagnosis, （2） efficacy of the ongoing antibiotic therapy, and （3） resolution of spontaneous bacterial peritonitis （SBP）.
METHODS： One hundred and twelve ascitic fluid samples were collected from 52 consecutive cirrhotic patients, 16 of them with SBP. The agreement between the manual and the automated method for PMN count was assessed. The sensitivity/specificity and the positive/negative predictive value of the automated blood cell counter were also calculated by considering the manual method as the ＂gold standard＂ RESULTS： The mean ＋ SD of the difference between manual and automated measurements was 7.8 4- 58 cells/ram3, while the limits of agreement were ＋124 cells/mm3 [95% confidence interval （CI）： ＋145 to ＋103] and -108 cells/mm3 （95% CI： -87 to -129）. The automated cell counter had a sensitivity of 100% and a specificity of 97.7% in diagnosing SBP, and a sensitivity of 91% and a specificity of 100% for the efficacy of the ongoing antibiotic therapy. The two methods showed a complete agreement for the resolution of infection.
CONCLUSION： Automated cell counters not only have a good diagnostic accuracy, but are also very effectivein monitoring the antibiotic treatment in patients with SBP. Because of their quicker performance, they should replace the manual counting for PMN determination in the ascitic fluid of patients with SBP.     

An update on peripheral blood progenitor cell transplantation

High-dose therapy with stem cell rescue is increasingly being used as a salvage or consolidation therapy for patients with poor-risk malignant disease. The availability of peripheral blood progenitor cells (PBPC) has opened new therapeutic perspectives to alleviate the severe toxicity related to prolonged myelosuppression. The preferred method of collection is still a matter of much debate. PBPC can be collected in steady state and after chemotherapeutic conditioning, growth factor priming, or both. Usually a heterogeneous population containing both committed progenitors and pluripotent stem cells can be harvested. Studies comparing engraftment after mobilized PBPC with recovery after autologous bone marrow transplantation confirm the beneficial effect on neutrophil and platelet engraftment. The accelerated hematological recovery can be associated with a number of clinical benefits including a reduction of platelet transfusions and shorter hospital stay. Only a few randomized studies are currently available on the long-term outcome after PBPC transplantation. Recent findings on tumor cell mobilization stimulated the development of techniques for tumor cell reduction, based on negative selection (purging) of tumor cells or positive selection of CD34-positive progenitor cells. Positive CD34 selection is also imperative for successful ex vivo expansion of progenitor cells and gene transfer experiments. The value of PBPC in the field of allogeneic transplantation is currently being examined.     

Use of the haemopoietic progenitor cell count of the Sysmex SE-9500 to refine apheresis timing of peripheral blood stem cells

The Sysmex SE-9500 automated cell counter provides an estimate of immature cells referred to as 'haemopoietic progenitor cells' (HPC). The aim of this study was to relate the HPC count to CD34+ cell levels in mobilized peripheral blood and to determine whether the HPC count was valuable in predicting apheresis yields of CD34+ cells. Studies were performed on 114 samples from 67 patients undergoing progenitor cell mobilization. HPC cells were undetectable in the steady state. On the day of apheresis the HPC and CD34 counts were weakly correlated, with the median HPC count being 2.3-fold greater than the CD34+ cell count. The HPC count did not include the CD34+ cells as immunomagnetic depletion of CD34+ cells did not significantly reduce the HPC count. CD34+ cell counts predicted for apheresis yield (r = 0.773) on that day as did the HPC count (r = 0. 623). The optimal strategy to prevent unnecessary harvesting while minimizing the risk of missing an adequate harvest, and minimizing laboratory investigations, was to screen all samples for HPC and limit CD34+ cell measurements to those with an HPC count <10 x 106/l (19/114 samples). If the CD34+ cell count was also <10 x 106/l then harvesting should not be carried out.     

Predictive parameters for mobilized peripheral blood CD34+ progenitor cell collection in patients with hematological malignancies

In order to investigate what is the best single parameter to predict the leukapheretic yield of circulating CD34+ progenitor cells, we retrospectively analyzed data from 68 patients with hematological malignancies who underwent mobilizing therapy. Three main parameters were monitored: total white blood cell (WBC), CD34+ cells, and monocyte counts in peripheral blood (PB) at the same day and at the preceding day of the apheretic procedure. Linear regression analysis revealed a strong correlation between CD34+ cell value in PB just before harvest and the number of CD34+ cells collected (P < 0.0001), but not at the preceding day. Monocyte PB concentration and absolute WBC count did not correlate with CD34+ cells harvested, at the preceding day of leukapheresis as well as at the same day of the procedure. The number of CD34+ cells in mobilized PB at the same day of harvest evidenced a very good capacity of predicting the value of harvested CD34+ cell number after collection, while WBC and monocyte count displayed quite a wide dispersion of results. In particular, an amount greater than 50/μL of circulating CD34+ cells ensured the best collections. Finally, CD34+ and CFU-GM content evaluated for each apheresis showed a strong reciprocal correlation (r 0.78; P < 0.0001). We conclude that the absolute number of CD34+ cells at the day of leukapheresis is the only parameter for identifying the exact timing for apheresis and predicting the amount of peripheral blood progenitor cells (PBPCs) that will be collected. In this setting, WBC and monocyte counts, at the day of collection or at the preceding day, are not useful tools. Am. J. Hematol. 58:255–262, 1998. © 1998 Wiley-Liss, Inc.     

Immunological characterization of canine hematopoietic progenitor cells

Summary Canine hematopoietic progenitor cells were characterized by separation with monoclonal antibodies. Depleted and enriched fractions were studied for growth of CFU-GM in semisolid agar and for repopulating capacity of lethally irradiated dogs. CFU growth was not reduced by depletion of marrow using monoclonal antibodies F 3-20-7 (anti-dog Thy-1), MT606 (anti-human CD6), and IOT2a (anti-human DR). CFU growth was variable following treatment with the anti-canine T-cell antibody MdT-P1 and immunomagnetic bead separation. It was regularly enriched when MdT-P1 treatment was followed by immunorosetting with staphylococcal protein A-loaded sheep red blood cells and density gradient separation. Lethally irradiated dogs were reconstituted by autologous marrow depleted of MdT-P1-positive cells using immunorosetting and density gradient centrifugation, whereas immunomagnetic bead-depleted marrow was ineffective. Fluorescence-activated cell sorting showed enrichment of hematopoietic progenitor cells in the weakly MdT-P1-positive fraction.Nonreviewed contribution to the annual meeting of the German and Austrian Societies of Hematology.This work was supported by the Wilhelm Sander Foundation, AZ 84.020.1This work is part of a doctoral thesis by J. Hahn, Ludwig-Maximilian-University Munich     

Circulating hematopoietic progenitor cells in polycythemia vera: The in vivo effect of hydroxyurea

The in vivo effects of hydroxyurea (HU) on circulating erythroid (BFU-E) and granulocyte-macrophage progenitors (CFU-GM) in patients with polycythemia vera (PV) have been evaluated. HU induced a strong decrease of both BFU-E and CFU-GM in the first month of treatment. During the following 4 months of treatment the level of circulating progenitors remained at very low values, until the end of the period of observation. HU activity involved both erythroid and myeloid committed progenitors and both erythropoietin-stimulated (normal) and endogenous (derived from the abnormal PV clone) BFU-E.     

FLT-3 ligand mobilizes hematopoietic primitive and committed progenitor cells into blood in mice

Abstract: We investigated the effects of the administration of FLT-3 ligand (FL) on mobilization of primitive and committed progenitor cells in mice. C57bI/6J mice were injected subcutaneously with FL once a day for 5 d at doses of 20, 100 and 200 μg/kg. After the collection of peripheral blood, we determined the number of white blood cells (WBCs) with the differential counts. The formation of colony-forming cells (CFCs) in peripheral blood, bone marrow and spleen was evaluated. Although the administration of FL, 20 μg/kg, did not stimulate leukocytosis, its administration at doses of 100 and 200 μg/kg increased the number of WBC up to 1.7- and 2.4-fold, respectively. Committed progenitor cells were mobilized into the peripheral blood dose-dependently and the number of CFCs was increased up to 5.5-fold by the administration of FL at 200 μg/kg on d 5. The number of CFCs in the bone marrow increased, but not dose-dependently. The number of CFCs in the spleen also increased up to 32-fold at a dose of 200 μg/kg FL. Mobilized peripheral blood mononuclear cells were transplanted into lethally irradiated mice and the number of CFU–S (d 12) was scored. A dose-dependent mobilization of CFU–S (d 12) into peripheral blood was also observed. These observations suggest that FL can mobilize hematopoietic primitive and committed progenitor cells into the peripheral blood of mice and those cells mobilized by FL may be applicable to peripheral blood stem cell transplantation.     

人巨细胞病毒对粒-单及红系祖细胞体外增殖的影响

目的：探讨人巨细胞病毒(HCMV)对脐血粒—单系祖细胞(CFU—GM)、红系爆式祖细胞(BFU—E)及红系祖细胞(CFU—E)体外增殖的抑制作用。方法：采用造血祖细胞体外半固体培养技术，研究HCMV—ADl69株对脐血CFU—GM、BFU—E及CFU—E集落生长的影响；用聚合酶链反应(PCR)技术检测集落细胞中HCMV—ADl69 DNA。结果：①HCMV—ADl69对CFU—GM、BFU—E及CFU—E均有抑制作用。②其抑制作用与HCMV浓度有关，高浓度(1：10，1：100)感染组与对照组比较集落产率明显下降(P＜0.01)，集落维持的时间明显缩短(P＜0.01)，而低浓度组(1：1000)无此作用。③经PCR检测发现病毒感染组CFU—GM、BFU—E及CFU—E集落细胞中有HCMV—ADl69 DNA存在。结论：①HCMV—ADl69可抑制CFU—GM、BFU—E及CFU—E集落的生长。②造血祖细胞是HCMV—ADl69的宿主细胞之一，HCMV—ADl69能直接感染造血祖细胞。HCMV感染患儿出现的粒细胞减少和贫血可能与此有关。     

Usefulness of routine admission complete blood cell counts on a general medical service

The usefulness of three components of the routine admission complete blood cell count (leukocyte count, hematocrit, and platelet count) was evaluated in 301 patients consecutively admitted to the internal medicine wards of a university teaching hospital. Using a consensus analysis approach, three faculty members reviewed the patients’ charts to determine which tests were performed routinely, which tests were abnormal, and which routine tests led to diagnostic or therapeutic changes. Overall, 55.3% of the tests were considered routine admission tests. Abnormalities were detected in 13.6% of the routine leukocyte counts, in 8.2% of the routine hematocrit levels, and in 12.4% of the routine platelet counts. However, treatment was changed for only three patients, all of whom had low hematocrits; this amounted to 0.6% of all tests. Furthermore, only one of the three patients received appropriate treatment that might have been withheld if a routine hematocrit determination had not been ordered. The authors conclude that the impact of routine admission complete blood cell counts on patient management is small and that the practice of ordering this test for all hospitalized patients could be eliminated with little adverse effect on patient care. Received from the Department of Medicine, Division of General Internal Medicine and Primary Care, University of California, Irvine, Medical Center, Orange, California, and the Department of Medicine, Division of Medical Education, MacNeal Hospital, Rush Medical School, Chicago, Illinois.     

红细胞单采在真性红细胞增多症治疗中的应用

目的:观察分析红细胞单采在真性红细胞增多症(PV)治疗中的应用方法及效果。方法:对符合国内PV诊断标准的19例患者,运用血细胞分离机进行异常增生红细胞单采,配合临床药物治疗。比较治疗前后患者血常规中Hb、RBC、Hct、WBC、PLT均值变化,采用SPSS17.0统计学软件统计分析。观察依患者公斤体重和Htc设定红细胞采集量,单次采集后Hct降低情况及首次采集前后Hb均值变化与再次采集前后Hb均值变化的差异。结果:治疗后血液Hb、RBC、Hct均值明显低于治疗前,差异有统计学意义(P0.01)。8例完全缓解,7例临床缓解,4例好转,有效率为100%;单次采集Hct平均降低幅度为16.11%。结论:红细胞单采在PV的治疗中具有显著的疗效,单次采集应考虑PV患者血容量绝对增加的情况,根据患者病情和对红细胞去除的耐受情况适当增加采集量。     

Peripheral blood progenitor cell (PBPC) transplantation with a single apheresis in patients with lymphoma,myeloma and solid tumors

Abstract: The aim of this study was to investigate if a single apheresis after peripheral blood progenitor cell (PBPC) mobilization can be used to rescue patients receiving high dose chemotherapy (HD.CHE) as treatment for an underlying malignancy. Eighteen consecutive patients who were admitted to the transplant unit for treatment were leukapheresed following mobilization with one of the following protocols: group I: rHuG–CSF alone, group II: conventional chemotherapy (C.CHE)+rHuG–CSF or rHuGM–CSF and group III: high dose Cytoxan (HD.CTX)+rHuG–CSF. The optimal day for leukapheresis was determined by following white blood cell counts (WBC), mononuclear cell counts (MNC) and CD34⁺ cell counts daily. Granulocyte – macrophage colony-forming cells (GM–CFC) assay was performed at the leukapheresis product and prior to reinfusion. All patients proceeded directly to ablative therapy according to their underlying malignancy. PBPC from single apheresis were reinfused to all patients and cytokines started 24 h after infusion. Hematologic recovery after HD.CHE was the parameter used to ensure successful engraftment. We have been able to recover adequate number of PBPC for transplantation with a single apheresis in all patients. The number of infused cells were for groups I, II and III: (1) median number of MNC 4.7, 3.58 and 2.79 × 10⁸/kg, respectively (2); median number of CD34⁺ cells 4.4, 2.8, 2.7 × 10⁶/kg, respectively. The median apheresis day was 6, 16 and 16, respectively. Recovery times to granulocyte count >0.5 × 10⁹/L was 9 d (range 9–12) and to platelets >20 × 10⁹/L was 12 d (range 1–135); 17/18 patients have engrafted successfully independent of the mobilization method used.These data suggest that sufficient PBPC can be harvested at a single leukapheresis for hemopoietic rescue after myeloablative therapy. Rapid hematologic recovery occurs when cytokines alone after conventional or HD.CHE are used for mobilization. Results of collection products and hematopoietic recovery are independent of the mobilization technique used.     

Evidences for age-related modulation of human hematopoietic progenitor cell proliferation

In order to identify hints of ageing in circulating hematopoietic stem cells (HSCs), putative senescence markers like the cellular level of carbonyl-modified proteins and senescence associated beta-galactosidase activity were measured. Furthermore, the number of HSCs in the periphery and their proliferative capacity in vitro were analyzed in buffy coats of fifty five individuals: 27 young [age, 19-43 years; mean age 31] and 28 middle-aged individuals [age, 45-66 years; mean age 56]. The effect of humoral factors on cell proliferation in culture was studied by expansion of the cells in the presence of plasma pools from children and elderly donors. Using a multiplex flow cytometry method, the plasma pools used in the proliferation experiments were assayed for the presence or absence of 25 chemokines. Within the age range analyzed, no age-dependent differences in the number of isolated CD34(+) cells were found. Both sources of progenitor cells were able to reach comparable cell density in culture, but cells from the middle-aged subjects proliferated only sufficiently in the presence of plasma obtained from older donors. Cells from middle-aged donors exhibited elevated levels of carbonyl-modified proteins and showed increased beta-galactosidase activity in comparison to the cells from young donors. Our study shows that although two markers of ageing i.e. carbonylated proteins and senescent associated beta-galactosidase activity are increased in HSCs from middle-aged donors, the number and proliferative capacity of HSCs are still maintained.     

Enrichment of hematopoietic progenitor cells from human bone marrow on percoll density gradients

Summary Hematopoietic progenitor cells were fractionated on continuous and discontinuous Percoll density gradients. It could be shown, that Percoll provides a suitable gradient medium for analytical as well as preparative density gradient procedures without major methodological problems. Advantages compared to albumin and Ficoll-Metrizoate are discussed.     

献血者定期一次性单采2U血小板的安全性分析

目的:探讨固定献血者定期一次性单采2U血小板的安全性。方法:按照《献血者健康检查要求》(GB18467-2011),研究52例单采献血者按每2周间隔时间且每次均采集血小板2U,分别于采后30min和14d留取献血者静脉血样进行血常规各项指标检测,同时考察单采6个月后免疫功能指标。结果:单采献血者采后30min除血小板相关指标(PLT、MPV、PDW、P-LCR)与采前比较差异有统计学意义(均P0.05),其他各项指标(WBC、RBC、Hb、Hct、MCV)则差异无统计学意义(均P0.05);采后14d留样观察与采前比较差异无统计学意义(均P0.05);连续单采6个月献血者的免疫性指标(IgG、IgM、IgA)及T淋巴细胞亚群的水平与捐献全血的献血者(n=60)之间差异无统计学意义(均P0.05)。结论:定期每2周一次性单采2U血小板能保证献血者的安全,且产品质量符合国家标准,能很好地缓解临床供需矛盾。     

HLA配型相合的造血干细胞移植治疗重型再生障碍性贫血的临床研究

目的 评价HLA配型相合的异基因造血干细胞移植(allo-HSCT)治疗重型再生障碍性贫血(SAA)的疗效.方法 2000年1月至2008年11月采用allo-HSCT治疗SAA患者20例,其中同胞相合移植17例,非血缘关系移植3例.预处理采用环磷酰胺(Cy)50 mg·kg~(-1)·d~(-1)4 d加抗淋巴细胞免疫球蛋白(ATG)2.5 mg·kg~(-1)·d~(-1)或20 mg·kg~(-1)·d~(-1) d.移植物抗宿主病(GVHD)的预防方案为经典的环孢素A(CsA)联合短程甲氨蝶呤(MTX)及霉酚酸酯(MMF).同胞供者采集经重组人粒细胞集落刺激因子(G-CSF)动员的骨髓及外周血干细胞,非血缘供者单纯采集外周血干细胞.结果 回输单个核细胞中位数为7.89(4.00-14.21)×10~(8)/kg,所有患者均获供者造血重建,粒细胞植活中位时间14(11～20)d;血小板植活中位时间12(8～108)d.但1例患者发生晚期排斥,行另一供者二次移植后植活.21例次移植后共发生6例次急性GVHD(I度3例,Ⅱ度皮肤3例),发生率16%.19例生存期>100 d的患者中有7例发生慢性GVHD,其中4例为局限型,3例为广泛型.截至2009年2月28 日,经过中位18(2.0～106.8)个月的随访,共有17例患者无病生存,总生存率为82.5%.结论 采用Cy+ATG的预处理方案对SAA患者进行HLA配型相合HSCT,植活率高,可以获得良好的疗效.