急性淋巴细胞白血病患者骨髓单个核细胞TAp63基因的表达

背景：据作者查新检索,国内外有关急性白血病患者骨髓单个核细胞TAp63基因表达的报道罕见。 目的：观察急性淋巴细胞白血病患者骨髓单个核细胞TAp63基因的表达及其意义。 方法：50例急性淋巴细胞白血病患者,其中32例急性B淋巴细胞白血病,18例急性T淋巴细胞白血病。同期选择27例非恶性血液病患者作为对照。取肝素抗凝的骨髓液2~4 mL,用Ficoll液分离骨髓单个核细胞,半定量反转录聚合酶链反应法检测TAp63的表达。 结果与结论：50例急性淋巴细胞白血病患者有49例表达TAp63,急性淋巴细胞白血病组明显高于非恶性血液病组(P < 0.05),急性B淋巴细胞白血病表达水平显著高于急性T淋巴细胞白血病(P < 0.05)。动态观察了5例初治急性淋巴细胞白血病化疗后不同阶段TAp63的表达变化,发现初治时TAp63表达,缓解后低表达或不表达,复发后又表达。结果表明TAp63在急性淋巴细胞白血病患者骨髓单个核细胞的表达明显高于非恶性血液病患者,尤其在急性B淋巴细胞白血病患者中高表达。     

Impact of bone marrow hematogones on umbilical cord blood transplantation outcomes in patients with acute myeloid leukemia

Early after umbilical cord blood transplantation, patients show marked differences in bone marrow (BM) hematogone percentages. Little is known about whether these differences are clinically relevant. We hypothesized that early recovery of hematogones may be associated with improved transplantation outcomes. BM aspirates were assessed from 88 patients with acute myeloid leukemia by two independent reviewers at day 21 and 100 after umbilical cord blood transplantation. Interobserver variability for BM hematogone percentages at these time points showed correlation coefficients of 0.83 and 0.98, respectively (P ≤ .01 for both). A high percentage of hematogones at day 21 was associated with less acute graft-versus-host disease grade 3 to 4 (P = .01). At day 100, a high percentage of BM hematogones was associated with improved overall survival (P = .02) and lower treatment-related mortality (P ≤ .01). This study shows that BM hematogone percentages may be useful prognostic indicators in patients with acute myeloid leukemia after umbilical cord blood transplantation and should be routinely reported in BM differential counts.     

A rapid, simple method for leukemia immunophenotyping using air-dried blood and bone marrow smears

This paper describes a modification of the peroxidase technique by which immunophenotyping may be carried out on routinely air-dried blood and bone marrow (BM) smears. The method is simple and quick, requires no special equipment, can be performed on fresh or stored specimens and gives a standard of morphological detail equal to that of routine blood films. With a monoclonal anti-HLA-DR antibody as a prototype, it was possible to demonstrate reliably, the presence of positively and negatively stained cells of appropriate morphological types in the peripheral blood of leukemia patients. Although only about one-third of antibodies tested were effective with the technique, we identified monoclonal antibodies capable of demonstrating myelomonocyte, granulocyte, monocyte, pan-leukocyte, transferrin, platelet, pan-T, 'cALLA plus B cell' and other antigens. However, we have not yet found antibodies able to identify T cell subsets, nor to distinguish 'common' acute lymphoblastic leukemia from its rare B-cell counterpart. With these limitations the method is suitable for routine use alongside cytochemistry in the differential diagnosis of leukemias and lymphomas.     

急性髓细胞性白血病患者外周血单个核细胞CD28 mRNA的表达

背景：大量研究显示,肿瘤患者外周血T细胞表面共刺激分子CD28蛋白表达存在差异,提示共刺激通路异常可能与恶性肿瘤的发生进展有关。 目的：观察急性髓细胞性白血病外周血单个核细胞共刺激信号分子CD28 mRNA在中的表达。 方法：急性髓细胞性白血病患者80例,其中M0型7例,M1型6例,M2型18例,M3型15例,M4型17例,M5型9例,M6型8例。并根据急性白血病疗效标准将80例患者分为完全治愈组、缓解组、未缓解组。采用Taqman探针实时荧光定量PCR检测80例患者及76名健康人群外周血单个核细胞CD28 mRNA的表达。 结果与结论：急性髓细胞性白血病外周血单个核细胞M1,M3和M4亚型中的CD28 mRNA表达量低于健康人群 (P< 0.05);急性髓细胞性白血病未缓解组中CD28 mRNA低于健康人群 (P < 0.05),完全治愈组和缓解组中CD28 mRNA表达与健康人群差异无显著性意义。说明急性髓细胞白血病患者外周血单个核细胞存在CD28 mRNA表达缺陷,并与临床分期、病情进展及预后有关。     

急性白血病患者骨髓细胞nucleostemin基因的表达与其类型、疗效的关系

目的：探讨nucleostemin（NS）基因在急性白血病中的表达及临床意义。方法:采用荧光定量PCR的方法检测67例急性白血病患者骨髓细胞nucleostemin基因表达，分析其表达与临床的关系。结果:初治急性白血病患者nucleostemin 基因的表达水平高于完全缓解组、正常对照组（P＜0.01），急性淋巴细胞白血病（ALL）患者nucleostemin表达水平低于急性非淋巴细胞白血病（ANLL）患者（P＜0.05），ANLL中M2、M3、M4、M5患者nucleostemin表达进行两两比较无显著差异（P＞0.05）,急性白血病患者nucleostemin表达水平与性别、年龄、肝、脾肿大和外周血白细胞数无明显相关（P＞0.05），nucleostemin 基因高表达的急性白血病患者的症状完全缓解（CR）率低于低表达的患者（51.3% vs 83.3%，P＜0.05）,急性白血病患者治疗达完全缓解者nucleostemin呈低表达，nucleostemin持续高表达或下降后又上升的患者呈现难治及复发。结论:Nucleostemin基因在急性白血病细胞中高表达，可作为疗效评价及监测残留病灶的指标。     

Clinical importance of myeloid-antigen expression in acute lymphoblastic leukemia of childhood

BACKGROUND. Leukemic cells in 15 to 25 percent of patients with acute lymphoblastic leukemia (ALL) express myeloid antigens as well as lymphoid antigens (the latter reflecting B-cell or T-cell lineage). The relations of myeloid-antigen expression to other features of ALL and to prognosis have been controversial. METHODS. We analyzed clinical and laboratory features present at diagnosis in 236 consecutive cases of ALL in children. Immunophenotyping, including single- and dual-fluorescence analyses, was used to classify leukemic cells as B or T lymphoblasts and also to identify myeloid-antigen expression--the simultaneous expression of lymphoid-associated antigens and at least one of three myeloid-associated antigens (CD33, CD13, and CD14) on cells classified as L1 or L2 according to the French-American-British system. RESULTS. Forty-five of 185 patients with B-lineage ALL had myeloid-antigen expression, as did 8 of 41 patients with T-lineage ALL. In 10 patients, the lineage could not be determined. Myeloid-antigen expression was associated with L2 morphology (P less than 0.05), but it did not correlate with other prognostic features recognized previously. Multivariate analysis showed that myeloid-antigen expression was an important predictor of relapse in childhood ALL and the most significant prognostic factor statistically (P less than 0.0001). A white-cell count greater than or equal to 50 x 10(9) per liter at diagnosis was also an important and highly significant prognostic feature (P less than 0.001). After 40 months, the estimated disease-free survival for patients with ALL was 84 percent for those without myeloid-antigen expression and with a low white-cell count, 57 percent for those without myeloid-antigen expression and with a high white-cell count, 47 percent for those with myeloid-antigen expression and a low white-cell count, and 26 percent for those with myeloid-antigen expression and a high white-cell count (P less than 0.00001). CONCLUSIONS. Myeloid-antigen expression is an important independent predictor of a poor response to chemotherapy in childhood ALL.     

Clinical importance of myeloid antigen expression in adult acute lymphoblastic leukemia

To determine the clinical importance of immunophenotypes in adult acute lymphoblastic leukemia (ALL), we prospectively studied 76 patients with this condition. Before treatment, lymphoblasts were tested for reactivity with monoclonal antibodies to B-cell, T-cell, and myeloid (My) antigens. Unexpectedly, myeloid antigens (MCS-2 or MY9) were identified in 25 patients (33 percent), usually in conjunction with B-cell or T-cell antigens. Among My+ patients, 15 (60 percent) expressed B-cell antigens (B+T-My+); all 6 tested had rearranged immunoglobulin genes. Five patients (20 percent) expressed T-cell antigens (B-T+My+), and one My+ patient expressed both B-cell and T-cell antigens. Only myeloid antigens (B-T-My+) were expressed in four patients (16 percent); three who were tested had germ-line immunoglobulin and T-cell-receptor gene configurations. Although no significant differences in presenting clinical features were found, My+ patients had fewer complete remissions than My- patients (35 vs. 76 percent, P less than 0.01). No differences in response or survival were observed between My+ and My- patients expressing T-cell antigens. However, among those expressing B-cell antigens, My+ patients had fewer complete remissions (29 vs. 71 percent, P = 0.02) and shorter survival (P = 0.03; median, 8.1 vs. greater than 26 months). These findings indicate that expression of myeloid antigen identifies a high-risk group of patients with adult ALL for whom alternative forms of treatment should be investigated.     

High correlation of the proteome patterns in bone marrow and peripheral blood blast cells in patients with acute myeloid leukemia

Background  

When comparing myelogenous blasts from bone marrow and peripheral blood, immunophenotyping usually show a strong correlation of expression of surface antigens. However, it remains to be determined, whether this correlation also exists on the level of protein expression.     

The comparison of bone marrow kinetics between patients with acute myeloid leukemia and acute promyelocytic leukemia after induction chemotherapy

Abstract

Background and Aim: Recently, acute promyelocytic leukemia (APL) has shifted from the most hazardous to the best curable type of acute myeloid leukemia. Anthracyclines, all-trans retinoic acid (ATRA) and arsenic derivatives are the most important developments for the treatment of APL. ATRA promotes the terminal differentiation of malignant promyelocytes to mature neutrophils. We aimed to compare platelet and neutrophil recovery time after induction chemotherapy in patients with acute myeloid leukemia (AML) and APL.

Materials and Methods: Two hundred and fifteen patients with AML and APL, who were diagnosed and treated in our tertiary care center between the years of 2001 and 2018 were evaluated.

Results: One hundred and eighty one AML patients (84.2%) and 34 (15.8%) APL patients were included in this study. The time between neutrophil nadir after induction chemotherapy and neutrophil recovery was longer in APL patients than in AML patients [30.5 (4–52) vs. 20 (5–58), p?<?0.001]. The time between platelet nadir after induction chemotherapy and platelet recovery was longer in APL patients than in AML patients [21.5 (4–42) vs. 17 (4–45), p?=?0.02].

Conclusion: Neutrophil and platelet recovery times were longer in APL patients than in AML patients in our present study. In 60?days, mortality rate was higher in APL patients than AML patients. Non-relapse mortality (NRM) rate was similar between two groups. There was a significant difference between two groups in terms of NRM causes. Platelet and neutrophil recovery time is very important because infection is the most important cause of NRM.     

急性早幼粒细胞白血病与MHC-A*基因的相关研究

目的探讨北方地区汉族人急性早幼粒细胞白血病（APL）与人类主要组织相容性复合体MHC～A＊基因多态性的关系。方法应用顺序特异性引物和聚合酶链反应（PCR—SSP）技术，对21例急性早幼粒细胞白血病患者及29例无血缘关系的健康人的MHC—A＊各等位基因及亚基因进行了检测分析。结果结果表明，MHC～A＊11（RR=5．333．P〈0．05=；和MHC—A＊24（RR=3．60，P〈0．05=基因与APL呈正相关，显著高于对照纽，两组之间比较差异有显著性意义，而其它MHC—A＊各等位基因未见异常，均无统计学差异。结论本项研究结果提示，MHC—A＊11和24基因可能是我国北方汉族人APL致病的易感基因，为揭示APL的发病机制中免疫遗传学作用提供了重要信息和依据。     

急性白血病患者骨髓细胞中TIMP3、RUNX3蛋白表达情况检测及其临床意义

目的： 检测急性白血病(AL)患者骨髓细胞中的TIMP3、RUNX3蛋白表达的情况及其与基因甲基化的关系,并探讨其与AL患者预后的关系。方法: 采用Western blotting检测50例AL患者骨髓单个核细胞(BMMCs)及10例健康供者外周血单个核细胞(PBMCs)中TIMP3、RUNX3蛋白表达的情况,结合前期实验中TIMP3、RUNX3启动子区域甲基化检测结果,对与AL患者预后有关的因素进行统计分析。结果: 50例AL患者BMMCs中RUNX3甲基化组该蛋白表达明显少于未甲基化组及正常对照组(P<0.05),AL患者完全缓解(CR)率与RUNX3蛋白表达及骨髓中原始细胞比例有关,RUNX3蛋白失表达、骨髓中原始细胞比例高的患者CR率低,反之较高。结论: RUNX3启动子区域甲基化是导致该蛋白失表达的原因,其参与了AL的发病,并与不良预后有关,TIMP3甲基化与急性白血病发病无关。     

Elevated IL-35 in bone marrow of the patients with acute myeloid leukemia

Acute myeloid leukemia (AML) is the most common hematological malignancy in adults, but the etiology of it remains poorly understood. IL-35 is a recently described cytokine composed of an IL-12 subunit p35 and an IL-27 subunit Epstein–Barr virus induced gene 3 (EBI3), and has an immunosuppressive effect on inflammation through induction of regulatory T cells (Tregs) and suppression of Th1 and Th17. Recently, we have illustrated that concentrations of IL-35 in peripheral blood are up-regulated in newly diagnosed (ND) AML patients. However, whether IL-35 in bone marrow is increased in AML patients is not clear. In this study, we examined IL-35 in bone marrow by various methods including RT-PCR, ELISA, FCM and IHC, and found that IL-35 levels are also increased significantly in bone marrow of adult AML patients. Furthermore, we investigated that concentrations of bone marrow IL-35 in ND group were higher than that in complete remission (CR) group and control group, but there was no significant difference compared to that in relapse group. In conclusion, IL-35 was elevated in bone marrow of adult AML patients and this increase was correlated with the clinical stages of malignancy, suggesting that IL-35 is involved in pathogenesis of AML.     

EvaGreen实时定量PCR检测急性髓系白血病GRAF基因表达

目的 研究急性髓系白血病(acute myeloid leukemia,AML)中粘着斑激酶相关性鸟苷三磷酸酶调节因子(GTPase regulator associated with the focal adhesion kinase,GRAF)基因的表达及其临床意义.方法 建立EvaGreen染料法实时定量聚合酶链反应(real-time quantitative PCR,RQ-PCR),对71例AML患者和21例良性血液病患者中GRAF转录本水平进行检测,分析其临床相关性.结果 所建立的EvaGreen RQ-PCR方法具有良好的特异性、重复性,敏感度达102拷贝/μL.AML患者中GRAF转录本水平(0.01%～169.75%,中位3.82%)较对照组(14.49%～126.85%,中位56.04%)显著降低(P<0.05),GRAF基因表达量与患者性别、年龄、外周血白细胞计数、血红蛋白、血小板计数、FAB亚型、染色体核型分组均无相关性(P>0.05).结论 AML患者中GRAF基因表达下调,可能与其发病有关.     

急性早幼粒细胞白血病与MHC-DRB1*基因的相关研究

目的探讨北方地区汉族人急性早幼粒细胞白血病(APL)与人类主要组织相容性复合体MHC—DRB1*基因多态性的关系。方法应用顺序特异性引物和聚合酶链反应(PCR-SSP)技术,对21例急性早幼粒细胞白血病患者及32例无血缘关系的健康人的MHC—DRB1*各等位基因及亚基因进行了检测分析,并将该方法与其它检测MHC等位基因的方法进行对比。结果结果表明,MHC—DRB1*09(RR=1.59,P<0.05=;MHC—DRB1*14(RR=1.59,P<0.05=基因与APL呈正相关,显著高于对照组,两组之间比较差异有显著性意义,而其它MHC—DRB1*各等位基因未见异常,均无统计学差异。结论本项研究结果提示,MHC—DRB1*09和14基因可能是我国北方汉族人APL致病的易感基因,为揭示APL的发病机制中免疫遗传学作用提供了重要信息和依据。