急性髓系白血病ERG、MDR1及BAALC基因的表达水平及临床意义

目的:探讨急性髓系白血病(acutemyeloidleukemia,AML)患者骨髓单个核细胞的ETS相关基因(ETS related gene,ERG)、多药耐药基因1(Multidrug resistance gene 1,MDR1)、脑和急性白血病胞质(Brain and acute leukemia cytoplasmic,BAALC)基因的表达水平及临床意义。方法:选取90例成人AML患者为研究对象,均接受蒽环类药物诱导化疗联合阿糖胞苷等进行初步治疗,采用实时荧光定量检测PCR技术检测治疗后的骨髓单个核细胞ERG、MDR1、BAALC基因表达,并分析其与患者临床特征、危险度分层、治疗疗效、生存率的关系。结果:AML患者ERG、MDR1、BAALC基因均有强表达,三因子表达强弱同白细胞、血小板等临床指标无明显相关性(P0.05),不同危险度分层对应的ERG、MDR1、BAALC表达之间有明显差异,中危组、高危组患者表达强度均高于低危组(P0.05)。ERG、MDR1低表达组对应的疗效CR(93.1%%,89.6%)、OS(56.5%, 66.7%)较高表达组CR(61.4%, 81.3%)、OS(56.5%,66.7%)值更高(P0.05),不同BAALC表达者疗效CR及生存率OS比较差异无统计学意义(P0.05)。结论:AML患者单个核细胞的ERG、MDR1基因表达水平与其危险度分层、疗效和生存率呈负相关,以REG和MDR1同AML关系敏感,BAALC敏感度较低,联合检测REG、MDR1、BAALC基因表达可能提高AML患者危险度分层、疗效及预后判读的准确度。     

网织红细胞在消化道肿瘤化疗前后的变化及临床意义

目的:探讨采用网织红细胞监测消化道肿瘤患者化疗前、后骨髓造血功能恢复的临床价值。方法:采用SysmexXE2100全自动细胞分析仪对60例消化道恶性肿瘤患者化疗过程中白细胞数(WBC)、血小板(PLT)、网织红细胞百分率(RET%)、未成熟网织红细胞指数(IRF)进行动态监测。结果:化疗后骨髓造血功能受到抑制,RET%和IRF从化疗后第3天开始出现明显下降,差异有统计学意义(P<0.01),在化疗后第14天开始出现回升,其中IRF值较第7天显著升高,两者间差异具有统计学意义(P<0.01);WBC和PLT则于化疗后第7天出现明显下降,差异具有统计学意义(P<0.01),化疗后第21天开始出现回升。结论:未成熟网织红细胞指数是预测化疗后骨髓造血恢复的较敏感指标。     

成人t(8;21)急性髓系白血病患者初诊Ki-67的表达特征及预后意义

目的:研究成人t(8; 21)急性髓系白血病(AML)初诊Ki-67抗原的表达特征及预后意义。方法:采集2012年7月至2019年2月本院57例成人初诊t(8; 21) AML患者的新鲜骨髓标本,采用流式细胞术(FCM)检测CD34和Ki-67抗原,分析Ki-67表达与患者初诊生物学特征、疗效及复发的关系。结果:全部患者中,CD34~+Ki-67~+细胞比例的中位值为30. 5%(范围:10. 0%～65. 8%);通过受试者工作特征(ROC)曲线确定CD34~+Ki-67~+细胞比例的最适分界阈值,CD34~+Ki-67~+细胞高比例与初诊c-KIT基因突变阳性及WT1转录本低水平均明显相关(P=0. 001; P=0. 042)。随访的36例患者中,CD34~+Ki-67~+高比例比低比例患者具有明显更高的1年累积复发(CIR)率(P=0. 035);此外,初诊WT1转录本低水平和微小残留病(MRD)高水平(2个疗程巩固治疗后RUNX1-RUNX1T1转录本水平下降3-log)均与更高的1年CIR率明显相关(P 0. 0001;P=0. 041),初诊c-KIT基因突变阳性和白细胞计数 10×109/L的患者分别有较高的1年CIR率趋势(P=0. 091; P=0. 054)。联合分组显示,MRD高水平同时CD34~+Ki-67~+细胞高比例的患者比其他患者具有明显更高的1年CIR率(P 0. 0001)。结论:初诊骨髓高比例的CD34~+Ki-67~+可能是成人t(8; 21) AML患者预后不良因素,MRD联合初诊CD34~+Ki-67~+细胞比例可能比单纯MRD更好地预测复发。     

急性髓细胞性白血病（AML）中IL-3Rα 基因的表达及意义

目的:检测白介素-3受体α亚基(IL-3Rα)在急性髓细胞性白血病(acute myeloid leukemia,AML)中的表达,研究其在AML发病和预后评估中的意义。方法:应用逆转录-聚合酶链反应(RT-PCR)法检测57例AML患者IL-3Rα的表达,其中包括52例初治和复发患者(6例M1、28例M2、10例M3、3例M4、5例M5)及5例缓解患者,另10例正常人作为对照。结果:①10例正常人和5例AML缓解患者未检测到IL-3Rα表达。②52例初治和复发的AML患者中IL-3Rα阳性表达21例,占40.38%,M1～M5各型患者的阳性率差异无显著性(P>0.05)。初治和复发AML患者之间IL-3Rα阳性表达无明显差异(P>0.05)。③AML患者的IL-3Rα阳性表达与外周血白细胞计数、骨髓中原始细胞比例、CD34阳性表达有关(P<0.05)。④AML患者中IL-3Rα阳性表达的患者CR率低于阴性者(P<0.05)。结论:①IL-3Rα在AML发病中有一定作用。②IL-3Rα基因可作为AML预后不良的一个指标。     

预激方案HAG及CAG治疗老年初治急性髓性白血病的对比研究

目的:评价含粒细胞集落刺激因子(granulocyte colony-stimulating-factor,G-CSF)的预激HAG及CAG方案治疗老年初治急性髓性白血病(AML)的疗效及不良反应,并对两个方案的疗效及不良反应进行比较。方法:65例老年初治AML分为HAG预激治疗组及CAG预激方案治疗组,31例患者予以HAG预激方案治疗,34例患者予以预激方案CAG方案治疗,所有患者在第1疗程后间歇14d左右进行第2个疗程。结果:HAG预激方案治疗组的完全缓解率为74.2%,总有效率为83.8%;CAG预激治疗组完全缓解(CR)率为67.6%,总有效率达为82.4%。两治疗组的血液系统不良反应及非血液系统毒性不良反应比较无显著性差异。结论:HAG预激方案化疗强度温和、敏感性好、CR率及有效率高、毒副作用小;与CAG预激治疗比较,HAG预激方案可以取的相似的疗效及较少的不良反应,在老年初治AML患者值得推荐应用。     

COVDP方案治疗成人急性淋巴细胞白血病疗效分析

目的:探讨不同诱导方案治疗成人急性淋巴细胞自血病的疗效及合并症情况。方法:采用COVDP及DVLP诱导治疗方案将我科(2005-2011年)62例急性淋巴细胞白血病患者分为两组,A组采用COVDP诱导方案,B组采用DVLP诱导方案。比较两种不同诱导方案治疗成人急性淋巴细胞白血病的初治缓解率、感染率、是否延迟化疗及骨髓抑制时间。结果:两组方案骨髓缓解率差异无统计学意义(P0.05),巩固治疗期间感染率差异有统计学意义(P0.05),A组骨髓抑制时间为(15.51±0.82d)长于B组(9.63±0.71 d)(P0.05),A组化疗延迟率为44.5%,高于B组(23.1%)(P0.05)。结论:与COVDP诱导方案相比,DVLP诱导方案缩短了骨髓造血恢复时间,降低了感染率及化疗合并症的发生。     

CD47 在急性白血病干细胞中的表达及其对临床疗效及预后的影响

目的:探讨CD47在急性白血病患者骨髓白血病细胞的表达及其临床意义。方法:选择2013年5月-2015年5月在我院确诊的急性白血病患者101例作为研究对象,其中急性淋巴细胞白血病50例(ALL组),急性髓系白血病51例(AML组)。另选取同期在我院接受体检的健康志愿者39例作为对照组。采用流式细胞仪检测白血病细胞表面CD47的表达情况,并分析CD47表达与急性白血病患者临床疗效及复发情况的关系。结果:急性白血病患者白血病细胞CD47的阳性表达率明显高于健康对照组,差异具有统计学意义(P0.05);而ALL组与AML组患者白血病细胞CD47的阳性表达率比较差异无统计学意义(P0.05);CD47阴性表达的急性白血病患者CR率显著高于阳性表达者,差异具有统计学意义(P0.05);ALL组和AML组CD47阴性表达患者CR率显著高于CD47阳性表达患者,差异具有统计学意义(P0.05),但两组之间比较,差异无统计学意义(P0.05);CD47阳性表达的急性白血病患者复发率显著高于阴性表达患者,差异具有统计学意义(P0.05);ALL组和AML组CD47表达阳性患者复发率明显高于阴性患者,差异具有统计学意义(P0.05),但两组之间比较差异无统计学意义(P0.05)。结论:急性白血病患者白血病细胞表面CD47的表达异常升高,且与白血病患者的疗效和预后有关,CD47可能作为一种急性白血病的诊断及疗效和预后的辅助评估指标。     

循环肿瘤细胞在胃癌化疗预后评估中的临床意义

目的:探讨检测循环肿瘤细胞对评估胃癌患者化疗敏感性和预后的临床价值。方法:选取行根治术的胃癌患者112例,术前留取血液标本进行循环肿瘤细胞检测。选取病理分期为Ⅲ期的36位患者进行术后6周期Xelox化疗,在化疗前和化疗结束后检测循环肿瘤细胞。分析循环肿瘤细胞检测值和肿瘤分期、化疗预后的关系。结果:在胃癌患者中,循环肿瘤细胞阳性率(2)为39.29%(44/112),并且和肿瘤分期与淋巴结转移明显相关(P0.05)。在术后行辅助化疗的胃癌患者中,循环肿瘤细胞阳性率和化疗敏感性明显相关,循环肿瘤细胞检测值动态增高的患者比降低的患者预后差(P0.05)。结论:循环肿瘤细胞的动态监测有助于预测胃癌患者的化疗预后。     

参芪扶正注射液对急性淋巴细胞白血病化疗患者造血和免疫功能的影响

目的:观察参芪扶正注射液对急性淋巴细胞白血病化疗患者造血及免疫功能的影响。方法:收集96例处于初治诱导缓解治疗阶段的急性淋巴细胞白血病患者,并将其随机分为治疗组和对照组。治疗组48例患者在进行常规化疗的同时给予参芪扶正注射液250 m L,1次/天,共28天;对照组48例患者仅接受常规化疗,两组患者均给予相同的支持对症治疗。治疗结束后,观察两组患者的疾病缓解率、治疗前后造血系统、T和B淋巴细胞亚群的变化情况。结果:化疗结束后,治疗组患者的疾病缓解率为89.6%,对照组为83.3%,两组比较无统计学差异(P0.05);化疗14天及化疗结束后1周,治疗组患者的白细胞、红细胞计数和血红蛋白水平明显高于对照组(P0.05);且化疗结束后1周,治疗组患者的CD3+、CD4+及CD4+/CD8+含量均明显高于对照组(P0.05)。而两组患者治疗前后的血小板计数、CD3-CD19+含量比较均不具有统计学差异(P0.05)。结论:参芪扶正注射液辅助治疗不仅能够改善化疗所致的急性淋巴细胞白血病患者的骨髓抑制,而且能够提高其细胞免疫功能,有助于患者化疗后的恢复。     

髓细胞性白血病患儿PTEN蛋白表达及其与免疫表型的关系

目的:探讨髓细胞性白血病(AML)患儿PTEN蛋白表达及其与免疫表型的关系。方法:选择AML患儿143例,根据免疫表型的不同分为两组:免疫分型为LY+AML型59例,LY-AML84例。所有患儿都给予免疫表型分析,检测PTEN蛋白表达情况并进行相关性分析。结果:LY+AML患儿CD34+阳性、CD117+阳性比例显著高于LY-AML患儿(P 0.05),染色体核型异常比例显著低于LY-AML患儿(P 0.05)。LY+AML患儿的PTEN蛋白表达量为(65.33±2.34)%,阳性表达率为94.9%;而LY-AML分别为(20.11±4.11)%和13.1%,与LY+AML患儿对比差异都有统计学意义(P 0.05)。在AML患儿中,Spearman相关分析显示PTEN蛋白表达水平与免疫分型呈现显著相关性(r=0.653,P=0.000)。多因素logistic回归方法显示PTEN蛋白表达、CD34+阳性、CD117+阳性、染色体核型异常为影响AML患儿免疫表型的主要独立危险因素(OR=1.098、1.045、1.092、0.294,P 0.05)。结论:AML患儿骨髓单个核细胞的PTEN蛋白表达上调使得LY+AML型发生风险显著增加,可作为AML患儿病情判断与预后预测的参考指标之一。     

Detection of minimal residual disease (MRD) after bone marrow transplantation (BMT) by multi-parameter flow cytometry (MPFC)

Multi-parameter flow cytometry (MPFC) was used to detect minimal residual disease (MRD) following bone marrow transplantation (BMT) in 21 patients. Bone marrow (BM) was analyzed pre-transplant and 3–4 months post-BMT while the patients were in clinical and morphological remission. MRD was detected by identifying cells with aberrant antigen expression and/or leukemia-associated phenotype (LAP) using MPFC. Prior to BMT, 8 out of 21 patients exhibited normal antigen expression based on normal BM samples while 13 BM aspirates had abnormal MPFC. Pre-BMT MPFC was abnormal in all 10 patients who were not in complete remission (CR) (>5% blasts in BM) as well as 3 patients acute lymphoblastic leukemia (ALL) who were in CR. In BM from ALL patients, an abnormal uniform B cell population was observed however antigen expression patterns varied greatly between patients. BM from acute myeloblastic leukemia (AML) patients showed an abnormal distribution of CD34+ cells. In addition, a correlation was observed between pre-BMT cytogenetics and MPFC. Only 2 out of 8 (25%) patients with normal MPFC pre-autologous bone marrow transplantation (ABMT) relapsed (AML), while 6 out of 13 (46%) patients with abnormal pre-BMT MPFC relapsed including 2 out of 3 patients who were transplanted in clinical CR. Pre-BMT MPFC may thus be an effective tool for detection of MRD by detection of a pre-transplant MPFC abnormality.     

Adoptive transfer of PR1 cytotoxic T lymphocytes associated with reduced leukemia burden in a mouse acute myeloid leukemia xenograft model

Background aimsTumor antigen-specific cytotoxic T lymphocytes (CTL) have been used in the treatment of human cancer, including leukemia. Several studies have established PR1 peptide, an HLA-A2.1-restricted peptide derived from proteinase 3 (P3), as a human leukemia-associated antigen. PR1-specific CTL elicited in vitro from healthy donors have been shown to lyse P3-expressing AML cells from patients. We investigated whether PR1-CTL can be adoptively transferred into NOD/SCID mice to eliminate human leukemia cells.MethodsPR1-CTL were generated in bulk culture from peripheral blood mononuclear cells (PBMC) stimulated with autologous dendritic cells. Human acute myeloid leukemia (AML) patient samples were injected and engrafted in murine bone marrow at 2 weeks post-transfer.ResultsFollowing adoptive transfer, bone marrow aspirate from mice that received AML alone had 72–88% blasts in a hypercellular marrow, whereas mice that received AML plus PR1-CTL co-infusion had normal hematopoietic elements and only 3–18% blasts in a hypocellular marrow. The PR1-CTL persisted in the bone marrow and liver and maintained a CD45RA^? CD28⁺ effector phenotype.ConclusionsWe found that adoptive transfer of PR1-CTL generated in vitro is associated with reduced AML cells in NOD/SCID mice. PR1-CTL can migrate to the sites of disease and maintain their capacity to kill the AML cells. The surface phenotype of PR1-CTL was consistent with their trafficking pattern in both vascular and end-organ tissues.     

Distribution of the cytoskeletal protein,Nestin, in acute leukemia

Abstract

Nestin is a neuroepithelial stem cell marker that is expressed in some types of tumor cells. Recent reports suggest that Nestin may be closely related to malignant cell proliferation and migration. Acute leukemia (AL) is characterized by a lack of differentiation, which results in uncontrolled proliferation in the bone marrow and accumulation of immature cells. The expression and function of Nestin in AL is unclear. We investigated Nestin immunohistochemical patterns of 87 patients that included 47 cases of acute myeloid leukemia (AML) and 40 cases of acute lymphoblastic leukemia (ALL), and 20 patients in complete remission (CR) from AML or ALL. We also investigated the clinico-pathological features of 87 cases of AL and their CR and overall survival (OS). Nestin was expressed in leukemic blasts and mature granulocytic cells in most cases (39/47) of AML. Conversely, Nestin was expressed in mature granulocytic cells in fewer cases (6/40) of ALL, but not in blasts. Nestin expression appeared in leukemic blasts of AML, but not ALL. Nestin expression in AML blast cells was not associated with CR or OS. We provide evidence that Nestin is expressed in AL and might be a useful immunohistochemical marker for identifying AML and ALL.     

Minimal Residual Disease-Based Risk Stratification in Chinese Childhood Acute Lymphoblastic Leukemia by Flow Cytometry and Plasma DNA Quantitative Polymerase Chain Reaction

Minimal residual disease, or MRD, is an important prognostic indicator in childhood acute lymphoblastic leukemia. In ALL-IC-BFM 2002 study, we employed a standardized method of flow cytometry MRD monitoring for multiple centers internationally using uniformed gating, and determined the relevant MRD-based risk stratification strategies in our local patient cohort. We also evaluated a novel method of PCR MRD quantitation using peripheral blood plasma. For the bone marrow flow MRD study, patients could be stratified into 3 risk groups according to MRD level using a single time-point at day-15 (Model I) (I-A: <0.1%, I-B: 0.1–10%, I-C: >10%), or using two time-points at day-15 and day-33 (Model II) (II-A: day-15<10% and day-33<0.01%, II-B: day-15≥10% or day-33≥0.01% but not both, II-C: day-15≥10% and day-33≥0.01%), which showed significantly superior prediction of relapse (p = .00047 and <0.0001 respectively). Importantly, patients with good outcome (frequency: 56.0%, event-free survival: 90.1%) could be more accurately predicted by Model II. In peripheral blood plasma PCR MRD investigation, patients with day-15-MRD≥10⁻⁴ were at a significantly higher risk of relapse (p = 0.0117). By multivariate analysis, MRD results from both methods could independently predict patients’ prognosis, with 20–35-fold increase in risk of relapse for flow MRD I-C and II-C respectively, and 5.8-fold for patients having plasma MRD of ≥10⁻⁴. We confirmed that MRD detection by flow cytometry is useful for prognostic evaluation in our Chinese cohort of childhood ALL after treatment. Moreover, peripheral blood plasma DNA MRD can be an alternative where bone marrow specimen is unavailable and as a less invasive method, which allows close monitoring.     

Effects of Decitabine on the proliferation of K562 cells and the expression of DR4 gene

Objective

To investigate the role of DR4 gene in the occurrence, development and prognosis of acute myeloid leukemia (AML), find a new regulatory gene of Decitabine for the treatment of AML, namely DR4 gene, and explore the molecular mechanism of AML in the treatment of AML.

Cytogenetic analysis and clinical significance of chromosome 7 aberrations in acute leukaemia

The analysis was performed on bone marrow cells derived from 96 patients with acute leukaemia (AL): 76 with acute myelogenous leukaemia (AML) and 20 with acute lymphoblastic leukaemia (ALL). Aberrations of chromosome 7 were revealed in 20 (21%) of 96 analysed cases: in 14 (18%) with AML and in six (30%) with ALL. Structural aberrations, present in 13 patients (eight with AML and five with ALL), were unbalanced and led to partial monosomy (12 cases) or trisomy (four cases) of chromosome 7. Twelve (86%) out of 14 AML and all the ALL patients with chromosome 7 aberrations had complex karyotypes in their bone marrow cells. Monosomy 7 and 7q losses were frequently observed in the AML group, whereas, in the ALL group, gains in 7q and losses in the short arms constituted most chromosome 7 aberrations. The occurrence of monosomy, or of losses in 7q, results in a worse response to induction therapy in AML patients. The complete remission (CR) rate was significantly lower in this group in comparison to the group of AML patients with a normal karyotype (p = 0.01) in bone marrow cells.     

Dosimetric predictors of acute bone marrow toxicity in carcinoma cervix — experience from a tertiary cancer centre in India

BackgroundThe objective of this study was To determine the dose volume parameters predicting acute haematological toxicity in carcinoma cervix patients undergoing concurrent chemoradiotherapy.Materials and methodsAll patients that presented to the hospital between Jan 2019 and Dec 2019 were prospectively analyzed. Patients diagnosed to have Carcinoma Cervix and planned for concurrent chemoradiation by volumetric modulated arc therapy (VMAT) were included for analysis. Patients were assessed at baseline and every week during treatment for acute haematological toxicities. Dose volume parameters from treatment plans were correlated with RTOG grade of haematological toxicities.ResultsA total of 34 patients diagnosed to have squamous cell carcinoma of cervix were treated by radical radiotherapy by VMAT technique and concurrent chemotherapy. The most common stage of presentation was stage IIB (61.7%). 29 patients (85.2%) completed five cycles of weekly cisplatin. Statistical analysis for sensitivity and specificity of dosimetric parameters was performed using receiver operating characteristic (ROC) curve. The probability of developing bone marrow toxicity was analyzed using T test. Mean dose to bone marrow exceeding 28.5 Gy was significantly associated with bone marrow toxicity (sensitivity — 82.4%, specificity — 70.6%). On analyzing dose volume parameters, volume of bone marrow receiving 20 Gy, 30 Gy and 40 Gy (V20, V30 and V40) more than 71.75%, and 49.75% and 22.85%, respectively, was significantly associated with bone marrow toxicity.ConclusionsOur study concludes that mean dose to bone marrow exceeding 28.5 Gy has high sensitivity and specificity for predicting bone marrow toxicity in patients receiving IMRT. Volume of bone marrow receiving 20 Gy, 30 Gy and 40 Gy significantly correlated with acute haematological toxicity.