骨髓增生异常综合征和再生障碍性贫血患者骨髓CD34+细胞测定

 目的 检测骨髓增生异常综合征（MDS）和再生障碍性贫血（AA）患者骨髓CD+34细胞占单个核细胞（MNC）的比率,以探讨二者可能的发病机制。方法 用流式细胞术（FCM）检测22例MDS患者、13例AA患者及12例非血液病患者骨髓CD+34细胞占MNC的比率。结果 AA组与对照组、AA组与MDS-RA组、AA组与MDS-RAEB组、MDS-RA组与MDS-RAEB组的骨髓MNC中CD+34细胞的比率的比较差异有统计学意义（P＜0.05）。大多数重型AA（SAA）患者（3／4）及很少慢性AA（CAA）患者（1／9）的骨髓MNC中的CD+34细胞的比率＜0.1 %。结论 骨髓CD+34细胞的检测有助于判断AA患者病情及MDS患者的预后,亦可用于鉴别AA和MDS。     

骨髓增生异常综合征患者骨髓单个核细胞CD34、CD117的表达及其临床意义

 目的　探讨骨髓增生异常综合征（MDS）患者骨髓单个核细胞CD34、CD117的表达及临床意义。方法　采用直接免疫荧光标记法标记细胞表面分化抗原,流式细胞术测定,对37例MDS患者的骨髓单个核细胞CD34、CD117表达及临床意义进行分析。依据MDS的WHO分型方案、染色体核型以及国际预后积分系统（IPSS）将MDS患者划分为RA-RARS-RCMD 组、RAEBⅠ-RAEBⅡ组;染色体良好组、染色体不良组;中危Ⅰ组、中危Ⅱ组、高危组。结果　RA-RARS-RCMD组19例中11例CD34、CD117表达阳性,RAEBⅠ-RAEBⅡ组18例患者CD34、CD117表达均阳性;随着疾病恶性程度的增高,CD34、CD117表达明显升高;染色体良好组22例中14例患者CD34、CD117表达阳性,染色体不良组15例患者均表达CD34、CD117;随着异常克隆的出现,CD34、CD117表达升高;中危Ⅰ组17例中9例CD34、CD117表达阳性;中危Ⅱ组11例、高危组9例,所有患者CD34、CD117表达均阳性;随着预后积分的递增,CD34、CD117表达随之升高。结论　CD34、CD117检测有助于MDS 的分型、预后判断。     

骨髓增生异常综合征患者骨髓单个核细胞CD34、CD117的表达及其临床意义

Objective To explore the expression and significance of CD34, CD117 on bone marrow mononuclear cells of myelodysplastic syndromes (MDS). Methods Direct immunofluorescence staining was used by means of flow cytometry. 37 patients with MDS were divided into RA/RARS/RCMD subgroup, RAEB Ⅰ/RAEB Ⅱ subgroup; favorable chromosomal subgroup, poor chromosomal subgroup; intermediate-risk Ⅰ subgroup, intermediate-risk Ⅱ subgroup, high-risk subgroup respectively according to WHO classification,cytogenetic abnormalities and international prognostic scoring system (IPSS). Results CD34 and CD117 were positive respectively in 11 of 19 patients with RMRARS/RCMD, all cases in RAEB Ⅰ/RAEB Ⅱ expressed CD34 and CD117; increased expression of CD34 and CD117 was MDS grade-related. Favorable chromosomal subgroup, 14 of 22 patients were positive for CD34, CD117, all cases in poor chromosomes expressed CD34 and CD117; there was a direct relationship between phenotytic density and poor cytogenetic risk factor. CD34 and CD117 expression was present respectively in intermediate-risk Ⅰ (9/17), intermediate-risk Ⅱ (11/11) and highrisk subgroup (9/9); the phenotypic intensity also was correlated with IPSS scores. Conclusion Detection of CD34, CD117 may be a useful tool for subtyping and predicting the prognosis of MDS.     

骨髓增生异常综合征与非造血干细胞克隆性疾病患者外周血及骨髓涂片细胞形态差异性研究

目的 对骨髓增生异常综合征(MDS)与非造血干细胞克隆性疾病患者外周血及骨髓涂片的细胞形态差异进行研究.方法 选取MDS患者与非造血干细胞克隆性疾病患者共165例,其中85例MDS患者作为观察组,80例非造血干细胞克隆性疾病患者作为对照组.采集观察组与对照组患者中骨髓细胞及外周血进行涂片,并分析和判定异常细胞比例.结果 观察组患者的血涂片中巨核细胞、幼红细胞、幼粒细胞及原始粒细胞比例均高于对照组,差异有统计学意义(P﹤0.05);观察组患者的棒状小体(Auer小体)、假性Pelger-Huet样核异常细胞、异常胞质颗粒、原粒细胞、核出芽、核空泡、核浆发育不平衡及高分叶的比例均高于对照组,差异有统计学意义(P﹤0.01);观察组患者多圆核巨核细胞、单圆核巨核细胞、畸形血小板、淋巴样小巨核比例均低于对照组,差异有统计学意义(P﹤0.01).结论 MDS中诊断价值最重要的是细胞形态学的异常,但部分病例在早期需要结合其他检测技术提高诊断的准确率.     

骨髓增生异常综合征中CD34+细胞CXCR4的表达及其与细胞迁移的相关性

  目的  探讨不同危险度骨髓增生异常综合征（myelodysplasticsymdromes,MDS）中骨髓CD34+细胞CXCR4的表达情况及其与细胞迁移率的相关性。   方法  收集40例骨髓增生异常综合征患者的骨髓标本,根据IPSS积分系统进行危险度分组。低危组20例:IPSS积分0~1.5分;高危组20例:IPSS积分≥1.5分;同时采集10例健康者的骨髓标本作为对照。分离纯化骨髓CD34+细胞,通过流式细胞术检测CXCR4膜蛋白的表达;研究SDF-1α趋化作用下CD34+细胞的迁移率及CD34+细胞对骨髓基质细胞的迁移率。   结果  高危组MDS患者CD34+细胞CXCR4的表达率明显高于低危组和正常对照组（P < 0.000 1）;低危组和正常对照组之间CXCR4的表达率无显著性差异（P>0.05）。高危组CD34+细胞对SDF-1α及骨髓基质细胞的迁移率显著高于低危组及正常组（均P < 0.000 1）,且其对骨髓基质细胞的迁移率与CXCR4的表达呈正相关（P=0.000 1）。   结论  高危组MDS患者CD34+细胞CXCR4的表达量及其对骨髓基质细胞的迁移率均明显高于低危组患者,且其迁移率随CXCR4表达量的增加而升高,不同风险组的MDS患者存在SDF-1及其受体CXCR4表达和功能上的差异,SDF-1及其受体CXCR4在MDS发病中具有重要作用。      

细胞发育异常在骨髓增生异常综合征患者预后评估中的价值

目的:探讨骨髓细胞病态造血在骨髓增生异常综合征(MDS)向急性白血病转化及预后评估中的价值。方法:回顾性分析82例MDS患者骨髓细胞形态学、细胞遗传学及流式细胞术等相关资料,分析患者骨髓细胞病态造血特点及患者预后转归。结果:82例MDS患者中,25例转化为急性髓系白血病(AML),其中45例存在原始细胞增多中20例转化为急性白血病,37例无原始细胞增多中5例转化为急性白血病,转化率差异有统计学意义(P=0.002);20例存在Auer小体者中13例转化为急性白血病,62例无Auer小体者中12例转化为急性白血病,两组转化率差异有统计学意义(P=0.000 1);31例小巨核病态改变者中15例转化为急性白血病,51例无小巨核病态改变者中10例转化为急性白血病,转化率差异有统计学意义(P=0.006)。其余病态造血未发现与转化为急性白血病有相关性(均P>0.05)。82例MDS患者中,27例死亡。其中存在原始细胞增多、Auer小体以及小巨核者,其生存时间较短。结论:原始细胞增多、Auer小体、小巨核病态改变与转化为急性白血病及其预后有相关性,对指导预后有一定意义。     

骨髓增生异常综合征14例误诊分析

对我院1995年1月—2002年11月首诊误诊的14例骨髓增生异常综合征(MDS)进行分析，以期提高对本病的认识，减少误诊误治。     

白血病,骨髓增生异常综合征患者外周血NK细胞活性的研究

用＾１２５ＩＵＤＲ释放法体外测定３０例白血病、１３例骨髓增生异常综合征（ＭＤＳ）和１８例再生障碍性贫血（再障、ＡＡ）患者外周血ＮＫ细胞活性。结果表明：三组ＮＫ细胞活性均明显减低，说明ＮＫ细胞活性降低在三种疾病的发病过程中起重要作用。进一步分析ＮＫ细胞活性与病情变化有关，故认为，定期检测ＮＫ细胞活性有助于患者病情监测、疗效及预后判断。     

112例骨髓增生异常综合征细胞遗传学分析

目的:探讨中国辽宁骨髓增生异常综合征(myelodysplastic syndrome,MDS)患者细胞遗传学异常特征。方法:回顾性分析2010年至2012年根据FAB及WHO标准诊断为MDS的112例患者。采用传统细胞遗传学(conventional cytogenetics,CG)及荧光原位杂交(fluorescence in situ hybridization,FISH)研究其分子遗传学改变。结果:112例患者中具有克隆性染色体异常46例,占41.1%。具有单独异常克隆40例(35.7%),两个异常克隆9例(8.0%),三个及以上异常克隆17例(15.2%);其中+8最常见,占26.8%。结论:染色体异常核型与WHO分型没有明显的相关性,但是复杂核型在RAEB-I及RAEB-II中更常见。     

High-level CD34 expression on megakaryocytes independently predicts an adverse outcome in patients with myelodysplastic syndromes

Expression of CD34 on mature-appearing megakaryocytes can be seen in various intrinsic bone marrow (BM) disorders as well as reactive bone marrows. In this study we investigate the clinical significance of CD34+ megakaryocytes in myelodysplastic syndromes (MDSs). Expression of CD34 on megakaryocytes was assessed on BM biopsies obtained from 202 patients with MDS. High-level (≥20%) CD34 expression on megakaryocytes was found in BM of 29 patients (14%). The expression of CD34 on megakaryocytes is correlated with severe cytopenia, higher numbers of myeloblasts, more frequent and higher risk cytogenetic abnormalities, and a shorter overall survival. Multivariate analysis indicated that the expression of CD34 on megakaryocytes could be a strong and an independent poor prognostic factor in MDS, with a hazard ratio of 2.53.     

CD34+ CD90+ cells and late hematopoietic reconstitution after autologous peripheral blood stem cell transplantation

Autologous peripheral blood stem cell transplantation (PBSCT) has been demonstrated to result in rapid, stable long-term engraftment. However, there has been considerable debate concerning the cells responsible for early and late hematopoietic reconstitution after PBSCT. Recently, CD34⁺ hematopoietic stem and progenitor cells have been clearly divided into two subpopulations by flow cytometry; namely undifferentiated pluripotent stem cells and differentiated committed progenitor cells. However, only a few studies have defined which subset contained in graft products might be the most predictive for late hematopoietic reconstitution after PBSCT. In this review, we present updated information regarding the relationships between the number of infused CD34⁺ cells or their immature subsets such as CD34⁺CD90⁺ cells and the late hematopoietic reconstitution after PBSCT, and discuss the threshold dose of CD34⁺CD90⁺ cells required for sustained long-term engraftment.     

外周血CD34 +细胞计数对外周血干细胞采集时机选择及结果的影响

目的:探讨外周血造血干细胞动员过程中外周血CD34 +细胞计数的变化趋势及其对采集时机选择和采集结果的影响。 方法:收集2012年4月至2017年3月在山西省肿瘤医院行自体外周血造血干细胞动员的62例血液病患者临床资料,采用化疗联合粒细胞集落刺激因子（G-CSF）的动员方案,监测外周血及采集物中白...     

骨髓增生异常综合征患者Dickkopf-3基因启动子区甲基化状态研究

目的 了解骨髓增生异常综合征(MDS)患者WNT/β-catenin信号转导通路中拮抗基因Dickkopf-3(Dkk3)的甲基化状态,初步探索其甲基化状态与MDS发生及疾病进展的关系.方法 应用甲基化特异性PCR(MSP)法对43例MDS患者的骨髓或外周血Dkk3基因启动子区甲基化状况进行检测,并以70例门诊普通患者的外周血检测结果作为对照,同时对患者进行随访.结果 43例MDS患者中检出7例(16.3％)Dkk3甲基化,其中5例(11.6％)为半甲基化状态,2例(4.7％)为完全甲基化状态,70例正常对照中1例(1.4％)为Dkk3甲基化,组间DKK3甲基化率差异有统计学意义(x2=8.93,P=0.005).7例Dkk3甲基化的样本中,2例来自骨髓,5例来自外周血,其中难治性贫血(RA)2例,难治性细胞减少伴多系异常(RCMD)1例,难治性贫血伴原始细胞增多(RAEB)4例.单因素分析示不同性别、染色体核型、样本来源(骨髓/外周血)、危险度(WHO分类的预后评分系统≤2分及＞2分)及老年组与非老年组间甲基化率差异均无统计学意义(均P＞0.05).在35例长期随访患者中,9例患者转化为急性髓系白血病(AML)(6例DKK3甲基化者中3例,29例DKK3非甲基化者中6例),疾病进展组与稳定组间甲基化率差异无统计学意义(P＞0.05);死亡12例中3例为Dkk3甲基化阳性,但甲基化与非甲基化组间生存率差异无统计学意义(P＞0.05).结论 MDS患者中Dkk3基因存在较高的甲基化修饰,这可能是MDS发病的分子机制之一.本组患者例数较少,尚未发现甲基化与临床预后间的相关性.     

Clinical implications of blast immunophenotypes in myelodysplastic syndromes

Myelodysplastic syndromes (MDS) are malignant disorders of hematopoietic cells. For many neoplasms, immunophenotype data of the neoplastic cells provide valuable information in clinical practice. However, the clinical values of immunophenotype data have not yet been firmly established for MDS. Since MDS blasts are not predominant in the bone marrow and peripheral blood, which makes reliable immunophenotyping of blasts difficult, we used a newly developed density-centrifugation reagent to generate blast-enriched MDS samples for phenotyping. The key findings of our study, which phenotyped blasts from 116 patients with MDS or acute leukemia transformed from MDS, were the following. (1) MDS blasts were usually CD34⁺CD38⁺HLA-DR⁺CD13⁺CD33⁺CD2^-CD3^-CD5^-CD8^-CD19^-CD20^- in flow cytometric analysis and often lacked myeloperoxidase in cytochemistry, regardless of the MDS subtype. (2) MDS blasts showed asynchronous expression of antigens (expression of both stem cell antigens and antigens of mature myeloid cells). (3) During disease progression of MDS, phenotypic clonal evolution (transition from blasts with a relatively mature phenotype to blasts with a more immature phenotype) occurred in at least some cases. (4) CD7-positivity was an independent variable associated with a short survival in MDS. Further studies of blast immunophenotypes will deepen our understanding of MDS and hopefully improve the clinical approach to these intractable disorders.     

脐带血CD+34细胞体外扩增优化条件的研究

 目的　分析比较不同细胞饲养层、不同刺激因子、脐血CD+34 细胞的纯度和含量等因素对脐血CD+34 细胞在体外扩增的影响,以筛选其体外扩增的优化方案。方法　采用正交实验,通过MTT法、细胞计数法及流式细胞分析技术测定脐血CD+34 细胞的增生情况;并采用以上方法及半固体培养体系对该细胞优化条件下的增生情况及集落形成能力进行检测。结果　优化条件下脐血CD+34细胞增生10倍,明显高于对照组的2.8倍（P＜0.01）,集落形成（CFU-C 36.67±6.11）也较对照组强（CFU-C 16.33±1.53）（P＜0.01）。结论　该培养系统能促进脐血CD+34 细胞体外扩增并能较好保持脐血CD+34 细胞的干细胞性质。     

The clonogenic potential of selected CD34+ cells from patients with MDS appear preserved when tested ex vivo

Our aim was to examine in 17 patients with MDS the effects of PMA activated and non-activated autologous lymphocytes on selected bone marrow CD34+ progenitors, in dose response studies. We used a double layer culture technique. Compared with controls, there was no difference in the colony growth promoting capacity of autologous PMA stimulated or unstimulated blood lymphocytes from MDS patients. In addition, similar to control studies, increasing numbers of lymphocytes, (0, 1 × 10⁵, 1 × 10⁶) led to a corresponding increase in the number of CFU-GM (p = 0.04). We conclude that MDS blood mononuclear cells have the ability to stimulate colony growth of autologous CD34+ cells while these selected progenitors show a proliferative capacity that is similar to normal when they are isolated from the bone marrow accessory cells.