用干扰素α（IFN-α）联合体外长期骨髓液体培养（LTBMC）净化16例慢性期Ph+慢性髓细胞白血病（CML）的骨髓，研究结果为：IFN-α1000IU/ml和2×106MNC/ml对LTBMC中CML和正常骨髓的非贴壁层细胞无明显影响，对CML培养3周后和正常骨髓培养第2-3周的非贴壁层CFU-GM有抑制作用，对CML和正常骨髓在LTBMC中的基质细胞层形成均有一定影响。在有完整染色体资料的13例患者中，加IFN-α组培养4周后有8例Ph+染色体完全消失，2例Ph+＜50%，1例Ph+60%，2例无变化，而对照组中只有5例完全消失，3例Ph+＜50%，1例Ph+65%，4例无变化，而且加IFN-α组Ph染色体的消失或减少时间较对照提前1周。结论：IFN-α1000U/ml与LTBMC联合对CL骨髓的净化有协同效应，两者联合可用于临床净化慢性期CML骨髓。

To investigate the clinically applicable conditions that support substantial expansion of both primitive and more mature hematopoietic cells of umbilical cord blood (UCB) for transplantation in adults, enriched CD34+ cells from 8 fresh UCB samples and 4 expanded UCB products were cultured in defined serum-free medium (QBSF-60) in the presence of a cytokine combination of SCF, Flt-3-ligand (FL), thrombopoietin (TPO), IL-3 for up to 2 weeks. Fresh medium with cytokines was supplemented or exchanged at day 4, day 7, and day 10. The proliferative response was assessed at day 7, day 10, and day 14 by evaluating the following parameters: nucleated cell (NC), clonogenic progenitors (colony-forming unit-granulocyte-macrophage [CFU-GM], burst-forming unit-erythrocyte [BFU-E], CFU-GEMM, and high-proliferative potential colony-forming cell [HPP-CFC]), immunophenotypes (CD34+ cells and CD34+ subpopulations), and LTCIC. Simultaneously numerical expansion of various stem/progenitor cells, including primitive CD34+ CD38- HLA-DR- subpopulation and LTCIC, CD34+ cells, and clonogenic progenitors to mature nucleated cells, were continuously observed during the culture. An average 103.32±71.37×106 CD34+ cells (range 10.12×106-317.9×106) could be obtained from initial 1.72±1.13×106 UCB CD34+ cells after 10-14 days cultured under the described conditions. Sufficient CD341 cells (＞50.0×106) for transplantation in adults would be available in all but one UCB collections after 10-14 days expansion. The expanded CD34+ cells sustained most of the in vitro characteristics of initial unmanipulated CD34+ cells, including clonogenic efficiency (of both primitive and committed progenitors), the proportion of CD34+ CD38-HLA-DR- subpopulation, and the expansion potential. Initial addition of IL-3 to the cocktail of SCF+ FL+ TPO had positive effects on the expansion of both primitive and, especially, the more mature hematopoietic cells. It accelerated the expansion speed and shortened the optimal culture time from 14 days to 10 days. These results indicated that our proposed shortterm culture system, consisting of QBSF-60 serum-free medium with a simple early acting cytokine combination of SCF+ FL+TPO, could substantially support simultaneous expansion of various stem/progenitor cell populations involved in the different phases of engraftment. It would be a clinically applicable protocol for ex vivo expansion of CD34+ UCB cells.

目的 探讨CD133（AC133）在急性白血病（AL）中的表达及其意义。方法 采用三色荧光流式细胞术测定76 例AL患者白血病细胞膜上CD133的表达；采用半定量逆转录2聚合酶链反应（RT-PCR）方法测定CD133mRNA表达。结果 ①正常对照和AL患者的CD133mRNA表达与CD133蛋白表达相一致，AL患者CD133表达水平显著高于正常对照。②AL患者CD133及CD133mRNA高表达阳性率分别为42.1%和46.1%。急性髓系白血病（AML）M3患者CD133均高表达阴性，AML和急性淋巴细胞白血病（ALL）的CD133高表达率分别为43.4%和38.1%，差异无显著性。AML-M4CD133高表达阳性率显著高于其它AML亚型，而T-ALL和B-ALL的CD133高表达率分别为20.0%和43.7%，差异也无显著性。③AML患者骨髓细胞CD133表达与CD34、HLA-DR显著相关，ALL患者骨髓细胞CD133表达与CD34无关。④CD133表达与细胞或分子遗传学异常、发病时外周血白细胞数、乳酸脱氢酶水平、多药耐药基因（mdr1）表达及年龄等预后因素无显著相关。⑤CD133高表达阳性组完全缓解（CR）率及总反应（OR）率低于高表达阴性组，但仅有CD34/CD133共高表达阳性组CR率低于阴性组（44.4% vs 71.4%，P<0.05），差异有显著性。结论 AL 患者骨髓细胞CD133表达高于正常对照；检测CD133表达可能有助于AL各亚型的鉴别及疗效判断；CD133/CD34共同高表达可能是AL的一个不良预后因素。

我们用体外细胞培养的方法，对10例急性非淋巴细胞白血病（ANLL）患者在诱导缓解及巩固1～3疗程后外周血粒/单祖细胞（PBCFU-GM）的动态变化进行了系统观察。结果表明：目前国内冶疗ANLL的两个主要方案即DA和HA方案对外周血干细胞（PBSC）均具良好扩增作用；采购PBSC的最佳时机是完全缓解巩固1疗程的造血恢复期；血小板和网织红细胞的恢复速度（日增值）是预测PBSC水平的简便可靠的临床参数，两者同时上升且日增值分别超过20×109/L和0.2%预示PBSC的高水平。

对30例急性白血病（AL）自体骨髓移植（ABMT）后造血功能恢复进行了单因素和多因素分析。移植后WBC＞1.0×109/L、ANC＞0.5×109/L、BPC＞20×109/L和胸骨髓增生活跃的中位时间分别为30，37，61和50天。白血病类型（急性髓细胞白血病）和骨髓细胞-196℃冻存是造血功能尤其是血小板延期恢复的主要因素（P＜0.05），而年龄、性别、缓解（CR）至ABMT间期、停化疗至采髓及移植的间期、预处理方案回输制服髓细胞量等对造血功能的恢复无明显影响（P＞0.1）。血小板的延期恢复造成输血及血小板量明显增加和住院时间明显延长（P＜0.01）。移植后90天BPC<20×109/L者早期复发的可能性较大（P=0.06）。