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目的检测骨髓增生异常综合征(MDS)、再生障碍性贫血(AA)和急性白血病(AL)患者骨髓单个核细胞(BMMNC)的细胞周期分布及CD34、Ki67抗原的表达。方法选取2009年6月至2010年6月在山东大学附属省立医院就诊的住院及门诊患者68例,MDS组30例,其中难治性贫血(RA)组18例,难治性贫血伴原始细胞增多(RAEB)组12例;AA组22例,其中急性重型再障组7例,慢性非重型再障组15例;AL组16例,其中急性髓系白血病(AML)组12例,急性淋巴细胞白血病(ALL)组4例。另外选取正常对照组18例(均为非血液病患者)。应用流式细胞仪(FCM),通过碘化丙啶细胞核染色及免疫荧光双标法,对其BMMNC的细胞周期分布及CD34、Ki67的表达进行检测。结果与正常对照组相比,MDS和AL组BMMNCs中细胞周期G0/G1期、造血干/祖细胞CD34+细胞、Ki67+细胞、CD34+Ki67+细胞、CD34+细胞中Ki67+细胞和Ki67+细胞中CD34+细胞比例均显著升高(P<0.05),而细胞周期S和S+G2/M期细胞比例均显著降低(P<0.05);AL组细胞周期G2/M期细胞比例亦显著降低(P<0.05)。RAEB组与RA组以及AL组与RAEB组相比,RAEB组和AL组的G0/G1期、CD34+、Ki67+、CD34+Ki67+、CD34+细胞中Ki67+和Ki67+细胞中CD34+细胞比例均显著升高(P<0.05),而S、S+G2/M期细胞比例均显著降低(P<0.05)。与正常对照组相比,AA组BMMNC中G0/G1、S、G2/M和S+G2/M期细胞比例的差异均无统计学意义(P>0.05),而其CD34+、CD34+Ki67+、CD34+细胞中Ki67+和Ki67+细胞中CD34+细胞比例均显著降低(P<0.05)。与AA组相比,RA组的G0/G1期、CD34+、Ki67+、CD34+Ki67+、CD34+细胞中Ki67+和Ki67+细胞中CD34+细胞比例均显著升高(P<0.05),而S和S+G2/M期细胞比例均显著降低(P<0.05)。结论本研究表明,AA不同于MDS与AL,MDS和AL具有相似的发病机制,并且BMMNC的细胞周期分布及增殖特性的差异可作为RA与AA的诊断和鉴别诊断指标。
Objective To detect the cell cycle distribution and the expression of CD34 and Ki67 in bone marrow mononuclear cells (BMMNC) from patients with myelodysplastic syndrome (MDS), aplastic anemia (AA) and acute leukemia (AL). Methods From June 2009 to June 2010, 68 inpatients and outpatients from the Provincial Hospital of Shandong University were enrolled. Thirty patients in the MDS group, including 18 patients with refractory anemia (RA) and 18 patients with refractory anemia 12 cases in AA group, including 7 cases in acute severe aplastic anemia group and 15 cases in chronic non-azoocapsy group; 16 cases in AL group, 12 cases in acute myeloid leukemia group (AML) Lymphoblastic leukemia (ALL) group of 4 cases. Another 18 cases of normal control group (all non-hematological diseases). Flow Cytometry (FCM) was used to detect the cell cycle distribution and expression of CD34, Ki67 in BMMNCs by propidium iodide nuclei staining and immunofluorescence double labeling. Results Compared with the normal control group, the percentages of CD34 + cells in GMN / G1 phase, CD34 + cells, Ki67 + cells, CD34 + Ki67 + cells, Ki67 + cells and Ki67 + cells in MDS and AL groups (P <0.05), while the proportion of cells in S and S + G2 / M phase decreased significantly (P <0.05). The percentage of cells in G2 / M phase of AL group decreased significantly (P <0.05). The percentage of CD34 + cells in the G0 / G1 phase, the CD34 +, the Ki67 +, the CD34 + Ki67 + and the Ki67 + cells in the RAEB group and the AL group was significantly higher than those in the RAEB group, the AL group and the RAEB group <0.05), while the proportion of cells in S, S + G2 / M phase were significantly decreased (P <0.05). Compared with the normal control group, there was no significant difference in the proportion of G0 / G1, S, G2 / M and S + G2 / M phases in BMMNC of AA group (P> 0.05) The percentage of CD34 + cells in Ki67 + and Ki67 + cells in CD34 + cells was significantly decreased (P <0.05). Compared with AA group, the percentage of CD34 + cells in G0 / G1 phase, CD34 +, Ki67 +, CD34 + Ki67 + and Ki67 + cells in RA group were significantly increased (P <0.05) M phase cells were significantly lower (P <0.05). Conclusions This study shows that AA differs from ALS, MDS and AL in the pathogenesis of Alzheimer’s disease. Differences in cell cycle distribution and proliferation characteristics of BMMNC can be used as a diagnostic and differential diagnosis index for RA and AA.