骨髓增生异常综合征的免疫表型特征分析

 【摘要】　目的　评价免疫表型在骨髓增生异常综合征（MDS）诊断中的价值。方法　采用流式细胞术对27例MDS患者的骨髓细胞进行免疫表型检测。结果　随着MDS疾病的进展,CD+34细胞比例逐渐升高,分别为：难治性贫血／环形铁粒幼细胞性难治性贫血（RA／RAS）7.43 %,难治性贫血伴原始细胞增多（RAEB）36.81 %,难治性贫血伴原始细胞增多转化型（RAEB-T）56.45 %,3组差异有统计学意义（F＝51.197,P＝0.000）,且各组间差异均有统计学意义（P＜0.05）;髓系抗原CD33、CD13、HLA-DR表达逐渐增高,CD14、CD15抗原表达随着疾病的进展逐渐降低,3组间差异有统计学意义（P＜0.05）;B淋巴细胞表面抗原CD19、CD10的表达随着疾病进展而降低;T淋巴细胞表面抗原CD7表达随着疾病进展而增高,分别为RA／RAS 2.63 %、RAEB 10.79 % 和RAEB-T 11.00 %,3组间差异有统计学意义（F＝10.439,P＝0.001）,其中RA／RAS组与RAEB组、RAEB-T组之间差异有统计学意义（P＝0.000,P＝0.001）。结论　检测MDS患者骨髓细胞的免疫表型有助于MDS的诊断、分型和判断预后,从而为治疗提供依据。     

骨髓增生异常综合征细胞形态学及细胞化学特征的研究

 目的　探讨骨髓增生异常综合征（MDS）的细胞形态学及细胞化学改变,为MDS提供形态学诊断依据。方法　对53例已确诊MDS患者的骨髓涂片进行瑞特-吉姆萨染色及铁染色,外周血中性粒细胞碱性磷酸酶（NAP）染色,分析骨髓细胞形态学特点。结果　53例MDS骨髓细胞粒、红、巨核三系均有不同程度的病态造血表现,分别有35、30、33例,骨髓各系列中出现病态造血频率较高的排列次序为粒系>巨核细胞系>红系,骨髓中细胞形态学病态造血改变类型的发生率比例较高的依次为单圆核巨核细胞29例（54.7 %）、假Pelger-Hu?觕t核28例 （52.8 %）、红系巨幼样变25例（47.2 %）等。46例（86.8 %）的MDS患者NAP染色积分及阳性率减低或为正常值下限。结论　多数MDS有两系以上的病态造血改变以及NAP染色积分及阳性率改变。     

骨髓增生异常综合征免疫表型的研究

目的：评价免疫表型测定在骨髓增生异常综合征(MDS)诊断及分型中的价值。方法：应用单克隆抗体方法对55例MDS患者进行免疫表型检测。结果：MDS患者髓系抗原表达明显增高,FAB亚型的抗原表达呈现规律性改变,随着RA向RAEB／RAEB-t转化,较早期的髓系抗原(CD33)逐渐增加,较成熟的CD15抗原和T-淋巴细胞抗原逐渐减少;同时在RAEB/RAEB—t阶段CD34^ 细胞数明显增高;较早期的骨髓细胞表面抗原(CD38、HLA-DR)在MDS表达明显增加。结论：免疫表型的检测对MDS更精确的诊断和分型有重要意义。     

骨髓涂片联合骨髓活检诊断骨髓增生异常综合征86例

 目的　探讨骨髓穿刺涂片与骨髓活检切片同步观察对诊断骨髓增生异常综合征（MDS）的临床意义。方法　对86例MDS患者采用骨髓抽吸-活检双标本一步法取材,同步观察其涂片和切片。结果　86例MDS患者骨髓穿刺涂片增生程度极度减低至减低30例（34.88 %）,活跃、明显活跃、极度活跃56例（65.12 %）,红系病态造血43例（50.00 %）,粒系病态造血32例（37.21 %）,巨核系病态造血22例（25.58 %）。骨髓活检切片的增生程度极度减低至减低15例（17.44 %）,活跃、明显活跃、极度活跃71例（82.56 %）;红系病态造血16例（18.61 %）,粒系病态造血52例（60.47 %）,巨核系病态造血56例（65.12 %）。86例中66例切片与涂片WHO分型诊断一致,符合率76.74 %。结论　骨髓涂片和活检在MDS的诊断和分型中各有优点,两者相互补充,二者同步观察更有利于提高MDS诊断和分型的准确性。     

骨髓增生异常综合征各亚型染色体核型分布与预后的关系

目的 探讨骨髓增生异常综合征（MDS）各亚型中的染色体核型分布特点及其与预后的关系。方法 回顾分析151例原发性MDS患者的染色体核型,比较各亚型中的染色体核型分布特点、国际预后积分系统（IPSS）评分、白血病转化率及死亡率等,并比较其在汉族与维吾尔族MDS患者中有无民族差异性。结果 所有患者核型异常检出率为55.0 %（83／151）,其中简单异常占53.0 %（44／83）,复杂异常占47.0 %（39／83）。伴多系病态造血的难治性血细胞减少症（RCMD）、原始细胞过多的难治性贫血（RAEB）-Ⅰ、RAEB-Ⅱ亚型中复杂异常的检出率明显高于难治性贫血（RA）、环形铁粒幼细胞增多的RA（RARS）亚型。核型异常涉及各条染色体,发生频率较高的染色体畸变依次为-5／5q-、-7／7q-、＋8、-20／20q-、-X／-Y、i（17q）、9p-／9q-、＋21等。IPSS评分在各亚型中差异有统计学意义（χ2＝117.802,P＜0.01）;高危组的核型异常检出率明显高于低危组和中危组（均P＜0.05）。随访151例患者白血病转化率和死亡率分别为25.2 %（38／151）和43.7 %（66／151）,核型异常者白血病转化率和死亡率明显高于核型正常者（均P＜0.05）。核型异常者白血病转化中位时间和生存中位时间均短于核型正常者。汉族与维吾尔族MDS患者各亚型分布、核型异常特点及白血病转化率、死亡率等方面差异均无统计学意义（均P＞0.05）。结论 染色体核型异常在MDS不同亚型中存在差异且与预后密切相关,是影响MDS患者病情进展及预后的重要指标,对MDS的正确诊断、病情监测及预后评估有重要意义。     

病态造血细胞与细胞遗传学改变对骨髓增生异常综合征诊断及分型的意义

 目的 探讨病态造血细胞与细胞遗传学改变对骨髓增生异常综合征（MDS）诊断及分型的意义。方法 对132例MDS患者行常规骨髓穿刺及外周血涂片瑞氏染色,观察MDS各亚型各系列细胞的病态造血特点;同时行染色体核型分析,并结合病态细胞与染色体核型异常改变,分析MDS各亚型与之关联。结果 1.以检出病态细胞≥0.10观察,其粒、红、巨核三系总检出率为43.4%,对RA+RARS（低危）、RCMD（中危）及RAEB（高危）三组进行比较,其病态粒细胞及病态巨核细胞≥0.10者,主要见于RCMD（P＜0.01）;病态红细胞≥0.10,主要见于RA+RARS（P＜0.01）。2.MDS染色体核型异常总检测率44%,其异常核型检出率虽RA及RARS组低于其它各亚型,但未显示统计学意义（P＞0.05）。3.病态细胞及染色体核型异常检出与MDS亚型间关系表现为：RA组核型异常且同时具有病态细胞≥0.10者占50%,RCMD组占76%,RAEB组占60.9%（P＜0.01）。结论：染色体核型异常同时具有病态细胞≥0.10者显示与MDS亚型有关联;密切监测其造血及细胞遗传学改变对确诊MDS有帮助。     

家族聚集性骨髓增生异常综合征/急性髓系白血病家系报道一例并文献复习

目的 探讨家族聚集性骨髓增生异常综合征/急性髓系白血病(MDS/AML)的诊断、临床特点、基因突变及治疗转归.方法 分析1例家族聚集性MDS/AML家系中兄弟患者的骨髓细胞形态学、免疫分型、细胞遗传学、基因突变,对其疗效和转归进行观察,并复习相关文献.结果 先证者在确诊MDS-原始细胞过多难治性贫血Ⅰ型(RAEBⅠ)4个月后进展为AML,其兄在确诊MDS-难治性血细胞减少伴多系病态造血3个月后进展为MDS-RAEBⅡ,生存期分别为5个月和8个月.结论 家族聚集性MDS/AML临床罕见,其诊断需要结合家族史、细胞遗传学、分子生物学等进行综合判断,预后差.     

骨髓增生异常综合征骨髓细胞淋巴系免疫标志及其增殖特性临床意义探讨

目的：探讨骨髓增生异常综合征(MDS)骨髓细胞淋巴系免疫标志及其增殖特性的变化和意义。方法：用流式细胞仪(FCM)分析了45例MDS患者和10例良性血液病患者(对照组)骨髓细胞淋巴系免疫标志及其增殖特性。结果：MDS组T细胞标志CD2、CD7。早期B细胞标志CD9。干／祖细胞标志HLA-DR阳性表达率明显高于对照组(P＜0.05)；RA和RAEB组分别与对照组比较，T系标志CD2、CD7，阳性表达率明显高于对照组(P＜0.05)，HLA-DR明显高于对照组(P＜0.01)，B系标志CD9、CD10、CD19，与对照组无差异(P＞0.05)。随病情进展CD2+表达率明显下降，HLA-DR~+表达率明显升高。在S+G2／M期申CD2+细胞所占比例RAEB较RA明显降低(P＜0.05)，CD7+细胞所占比例RAEB较RA明显升高。结论：MDS骨髓细胞T淋巴系免疫标志和增殖特性异常，表明T淋巴细胞增殖分化受阻，提示细胞免疫功能失调在MDS发病和病情进展中可能起作用。     

异常核型骨髓增生异常综合征64例预后分析

 【摘要】　目的　探讨骨髓增生异常综合征（MDS）患者染色体异常与预后的关系,对治疗效果进行分析。方法　回顾性分析122例MDS患者染色体核型,用吉姆萨显带法进行检测。难治性贫血（RA）、环形铁幼粒细胞难治性贫血（RAS）的治疗以诱导分化剂及刺激造血药物为主。原始细胞过多难治性贫血（RAEB）、转化型原始细胞过多难治性贫血（RAEB-t）、慢性粒-单核细胞白血病（CMML）的治疗以小剂量化疗和小剂量联合化疗方案为主。分析异常核型MDS患者疗效,以同期住院的正常核型MDS患者为对照。结果　检出异常核型MDS患者64例,治疗后完全缓解（CR）17例,CR率26.6 %。同期正常核型MDS患者58例,CR 30例,CR率51.7 %。正常和异常核型患者CR率差异有统计学意义（χ2＝8.13,P＝0.04）。复杂核型、-7、+8核型异常者易进展为急性白血病。结论　染色体核型分析在MDS的诊断与预后判断中有重要意义,不同的染色体核型改变进展为白血病的风险不同。     

20q-骨髓增生异常综合征的临床特征和细胞遗传学分析

 目的　分析伴20号染色体长臂部分缺失（20q-）的骨髓增生异常综合征（MDS）患者的临床和染色体核型特征。方法　对10例伴20q-的MDS患者的临床表现、实验室检查、染色体改变及病程转归进行总结分析。结果　伴20q-的MDS多表现为三系血细胞减少,骨髓增生活跃或明显活跃9例（90 %）,以红系和粒系病态造血常见,10例伴有20q-的MDS中单纯20q-异常8例（80 %）,难治性血细胞减少伴有多系发育异常（RCMD）6例,难治性贫血伴有原始细胞过多-Ι（RAEB-Ι）2例,2例伴复杂核型的患者均为难治性贫血伴有原始细胞过多-Ⅱ（RAEB-Ⅱ）;2例患者转化为急性髓系白血病（AML-M1和AML-M2a）。结论　20q-可能是血液肿瘤中一种早期和初步的细胞遗传学改变,伴20q-的MDS以三系血细胞减少和病态造血常见,大多为低危组MDS,附加异常常累及5、7、8、14和17号染色体;单纯20q-比合并其他核型异常者生存期长。     

Dlk1 in normal and abnormal hematopoiesis.

Dlk1 (Pref-1) is a transmembrane and secreted protein, which is a member of the epidermal growth factor-like family, homologous to Notch/Delta/Serrate. We have found by real-time RT-PCR that Dlk1 mRNA levels were high in CD34(+) cells in 10 of 12 MDS samples compared with CD34(+) cells from 11 normals. Also, Dlk1 mRNA was elevated in mononuclear, low density bone marrow cells from 11/38 MDS patients, 5/11 AML M6 and 2/4 AML M7 samples. Furthermore, 5/6 erythroleukemia and 2/2 megakaryocytic leukemia cell lines highly expressed Dlk1 mRNA. Levels of Dlk1 mRNA markedly increased during megakaryocytic differentiation of both CMK megakaryoblasts as well as normal CD34(+) hematopoietic stem cells. High serum levels of Dlk1 occurred in RA (4/10) and essential thrombocythemia (2/10) patients. Functional studies showed that forced expression of Dlk1 enhanced proliferation of K562 cells growing in 1% fetal bovine serum. Analysis of hematopoiesis of Dlk1 knockout mice suggested that Dlk1 contributed to granulocyte, megakaryocyte and B-cell clonogenic growth and was needed for generation of splenic B-cells. In summary, Dlk1 is overexpressed in selected samples of MDS (especially RA and RAEB) and AML (particularly M6, M7), and it appears to be associated with normal development of megakaryocytes and B cells.     

骨髓增生异常综合征细胞形态学与荧光原位杂交检测染色体异常的相关性

目的：总结分析骨髓增生异常综合征（MDS）细胞形态学异常与异常克隆的相关性。方法回顾性分析山西医科大学第二医院血液科120例临床诊断为MDS且荧光原位杂交（FISH）检测及形态学资料完整的病例,总结MDS常见的核型异常（-5／5q-、-7／7q-、20q-、+8、-Y及复杂核型）与细胞形态学异常间可能的相关性。结果FISH检测发现克隆性染色体异常62例（51.7%）。核型异常组中5q-、-5及复杂核型组巨核系发育异常明显,单圆核巨核细胞[5q-组为87.5%（7／8）,-5组为100.0%（2／2）,复杂核型组为83.3%（5／6）]及小巨核细胞[5q-组为75.0%（6／8）,-5组为100.0%（2／2）,复杂核型组为66.7%（4／6）]检出率明显增高,与其余核型组检出率比较,差异有统计学意义（P＜0.05）。各异常核型组粒系及红系形态学发育异常检出率差异均无统计学意义（均P＞0.05）。高危组、中危组、低危组核型异常检出率分别为100.0%（4／4）、53.9%（55／102）、21.4%（3／14）,高危组与低危组间、低危组与中危组间核型异常比例差异有统计学意义（P＜0.05）。结论异常克隆与形态学改变可能相关,其中5q-、-5及复杂核型的形态学发育异常多表现在巨核系细胞中。     

Association between phenotypic features of blasts and the blast percentage in bone marrow of patients with myelodysplastic syndromes

Although the blast percentage in the bone marrow (BM) is a key parameter for the classification of myelodysplastic syndromes (MDS), the current blast percentages used to define MDS subtypes have not been shown to have strong biological relevance. We determined the blast phenotypes and examined their relationship with the BM blast percentage in 90 MDS cases. When the BM blast percentage increased, cases whose blasts expressed CD7, CD56 and CD117 increased whereas cases whose blasts expressed CD10, CD11b and CD15 decreased. The BM blast percentages where the blast immunophenotype changed were 5, 10, 20 and 25%. Blast immunophenotypes have the potential to provide a biological basis for and refine the present MDS classifications.     

汉族与维吾尔族三阴性乳腺癌临床特征及预后差异分析

探讨汉族与维吾尔族三阴性乳腺癌患者临床特征及预后,了解两民族三阴性乳腺癌临床特征的差异。方法：选取2003年1月至2005年1月新疆肿瘤医院收治的可手术切除并经病理证实的458例汉、维族乳腺癌患者,将ER、PR、Her-2表达均阴性的患者,分为汉族及维族两组,比较两组临床特征、复发转移情况及5年无瘤生存率。结果：458例乳腺癌患者当中,其中97例维吾尔族患者,361例汉族患者,三阴性乳腺癌总例为108例,所占比例为23.6%（108/458）。108例三阴性乳腺癌患者中,维吾尔族患者28.9%（28/97）,汉族患者为22.2%（80/361）。两组在肿瘤大小及临床分期方面差异有统计学差异（P<0.05）,但两者的复发转移模式及5年无瘤生存率无明显差别。结论：维吾尔族三阴性乳腺癌患者就诊时较汉族患者偏晚,T3期所占比例要高于汉族患者,T1期比例低于汉族患者,同时Ⅲ期患者比例明显高于汉族患者。但两组5年无瘤生存率无差别,应采取进一步研究以明确影响汉、维族三阴性乳腺癌患者的预后因素。     

Mesenchymal stem cells in myelodysplastic syndromes: phenotypic and cytogenetic characterization

Bone marrow-derived mesenchymal stem cells (MSC) have been defined as primitive, undifferentiated cells, capable of self-renewal and with the ability to give rise to different cell lineages, including adipocytes, osteocytes, fibroblasts, chondrocytes, and myoblasts. MSC are key components of the hematopoietic microenvironment. Several studies, including some from our own group, suggest that important quantitative and functional alterations are present in the stroma of patients with myelodysplasia (MDS). However, in most of such studies the stroma has been analyzed as a complex network of different cell types and molecules, thus it has been difficult to identify and characterize the cell(s) type(s) that is (are) altered in MDS. In the present study, we have focused on the biological characterization of MSC from MDS. As a first approach, we have quantified their numbers in bone marrow, and have worked on their phenotypic (morphology and immunophenotype) and cytogenetic properties. MSC were obtained by a negative selection procedure and cultured in a MSC liquid culture medium. In terms of morphology, as well as the expression of certain cell markers, no differences were observed between MSC from MDS patients and those derived from normal marrow. In both cases, MSC expressed CD29, CD90, CD105 and Prolyl-4-hydroxylase; in contrast, they did not express CD14, CD34, CD68, or alkaline phosphatase. Interestingly, in five out of nine MDS patients, MSC developed in culture showed cytogenetic abnormalities, usually involving the loss of chromosomal material. All those five cases also showed cytogenetic abnormalities in their hematopoietic cells. Interestingly, in some cases there was a complete lack of overlap between the karyotypes of hematopoietic cells and MSC. To the best of our knowledge, the present study is the first in which a pure population of MSC from MDS patients is analyzed in terms of their whole karyotype and demonstrates that in a significant proportion of patients, MSC are cytogenetically abnormal. Although the reason of this is still unclear, such alterations may have an impact on the physiology of these cells. Further studies are needed to assess the functional integrity of MDS-derived MSC.     

汉族和维吾尔族三阴性乳腺癌患者组织中E-cad及VEGF的表达差异

目的 分析汉族及维吾尔族女性三阴性乳腺癌（TNBC）患者组织中E-钙黏蛋白（E-cadherin, E-cad）、血管内皮生长因子（vascular endothelial growth factor, VEGF）的表达情况,了解其与预后间的关系。方法 选择新疆医科大学附属肿瘤医院2008年3月—2010年3月汉族TNBC患者140例,维吾尔族57例,检测E-cad、VEGF的表达情况,并与淋巴结转移及TNM分期、组织学分级进行比较,同时观察两组5年无瘤生存率。结果 两组TNBC患者5年无瘤生存率差异无统计学意义（P>0.05）;E-cad的表达无差别,VEGF阳性率汉族低于维吾尔族（P<0.05）,同时,两组E-cad的表达与淋巴结转移、TNM分期及组织学分级呈负相关（-1≤r<1, P<0.05）,而VEGF的表达与淋巴结转移及TNM分期呈正相关（0P<0.05）,但与组织学分级没有明显关系。结论 E-cad在汉族与维吾尔族TNBC中没有表达差异,VEGF阳性率在汉族女性TNBC中低于维吾尔族,但两组5年无瘤生存率没有明显差别,提示维吾尔族TNBC患者可能存在与汉族患者不同的影响其预后的因素,需采取进一步研究证实。     

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.