原发骨髓增生异常综合征患者IPSS及染色体核型的临床分析

目的:比较原发性骨髓增生异常综合征(MDS)患者WHO(2001)分型与FAB分型的IPSS染色体核型分析及预后的相关性分析。方法:经FAB标准确诊的原发MDS的患者重新按WHO标准分型,对2种结果的IPSS及染色体异常与各亚型的关系进行分析。结果:按FAB分型各亚型的IPSS及染色体异常无显著性差异,按WHO分型的难治性细胞减少伴多系增生异常(RCMD)与难治性贫血(RA)患者染色体异常率有统计学意义(66.6%,41.7%,P<0.01),RAEB-2高危组比例明显高于RAEB-1组(25%,0%,P<0.01)。结论:原发MDS的WHO分型与FAB分型相比,前者与预后的相关性更好。     

急性白血病细胞bcl-2、c-myc和p53基因表达的研究

目的研究bcl-2、c-myc、p53基因在急性白血病(AL)细胞中的表达及其与AL的分型、临床特征、疗效及预后因素的关系.方法采用免疫组化方法检测54例初治AL骨髓细胞bcl-2、c-myc、p53蛋白表达情况,分析其与FAB分型、临床特征、疗效及预后因素的关系.结果AL骨髓细胞中bcl-2、c-myc及p53基因的表达显著高于正常对照组(P<0.05).bcl-2、c-myc、p53基因表达在急性髓系白血病(AML)与急性淋巴细胞白血病(ALL)差异不显著;在AML亚型中,bcl-2在M4及M5中的表达高于M1、M2和M3(P<0.05);而c-myc及p53的表达在各亚型中差异无显著性意义(P>0.05).bcl-2的表达与初诊时的白细胞数呈正相关(r=0.44,P<0.05),与疗效呈负相关(r=-0.40,P<0.05),与 c-myc的表达呈正相关(r=0.51,P<0.05);c-myc 的表达与初诊时的白细胞数呈正相关(r=0.50,P<0.05).结论bcl-2、c-myc、p53基因的紊乱可能在AL的发病中起着重要作用,还与白血病的某些临床特征、疗效及预后因素密切相关.     

形态学、免疫学、细胞遗传学联合检测诊断急性白血病的临床意义

为探讨形态学、免疫学、细胞遗传学联合检测对急性白血病(AL)诊断、治疗、预后判断等方面的临床意义，对初诊为AL的39例患者分别进行了骨髓细胞形态学、免疫学及染色体检测。并按照FAB标准进行形态学(组织化学染色)分型；采用间接免疫荧光法标记活细胞膜表面分化抗原(CD)进行免疫学分型；采用24小时培养法制备染色体标本，G带显示法进行染色体检查。结果：39例患者经形态学检查确诊为AL，其中急性淋巴细胞白血病(ALL)8例，急性非淋巴细胞白血病(ANLL)29例，2例难以分型；免疫学诊断为ALL8例(其中2例伴髓系抗原表达)，ANLL29例(其中4例伴淋巴细胞抗原表达)，2例形态学难以分型者，诊断为急性杂合性白血病。免疫学与形态学分型符合率94．9％(37／39)。39例中染色体核型异常18例。本研究结果还显示，经临床治疗后染色体复杂畸形者缓解率低，正常核型及某些染色体核型[如t(15；17)]者缓解率较高。认为形态学、免疫学、细胞遗传学联合检测可提高AL诊断的准确性，有助于制定治疗方案及判断预后。     

儿童急性白血病形态学免疫学和细胞遗传学的分型诊断

目的：准确地进行儿童急性白血病（AL）的诊断分型,提高初诊患儿的诊断符合率。方法：采用形态学、免疫学和细胞遗传学（MIC）相结合的诊断方法,分析了110例初诊为AL的患儿。结果：形态学与MIC分型诊断符合率为88．2％;急性淋巴细胞性白血病（ALL）免疫分型诊断符合率为92．2％;而急性髓细胞性白血病（AML）仅为62．9％。8／35例AML表达淋系抗原（1y^ -AML),12／59例ALL表达髓系抗原（My^ -AML);11／110例为杂合性白血病。染色体核型异常检出率为63．6％。t(8;21）易位见于（13／21例）M2;t(7;11）易位见于12例M2;t（15;17）易位见于（2／5例）M3;t（9;22）和t（4;11）易位见于（8／64例）ALL。结论：运用MIC诊断分型方法能提高儿童AL的诊断率,为AL个体化治疗和评估预后提供信息。     

MLL基因重排在急性白血病亚型分布及疗效分析

目的:分析MLL基因重排急性白血病的临床、实验室特征及预后情况。方法:回顾性分析2016年8月—2020年8月确诊的23例MLL基因异常急性白血病患者的临床和实验室资料。结果:2016年8月—2020年8月我院收治的急性白血病患者共350例,其中急性髓系白血病(AML)225例,急性淋巴细胞白血病(ALL)125例,有MLL基因重排者23例(AML-M5 7例,AML-M4 3例,MDS转化AML 4例,AML-M2 4例,B-ALL 5例),占所有急性白血病的6.57%(23/350),在AML中比例为8%(18/225),在ALL中比例为4%(5/125)。18例MLL基因重排AML患者中,MLL-PTD阳性11例(61.1%),MLL-AF6阳性1例,MLL-ELL阳性3例,MLL-AF9阳性3例。5例MLL基因重排B-ALL患者,均为MLL-AF4阳性。23例患者经过2个疗程诱导化疗后,只有11例(47.8%)达完全缓解。接受移植的9例患者中位生存期为24(12～45)个月,未接受移植的14例患者中位生存期为15(0.53～48)个月,差异有统计学意义(P0.05)。结论:伴MLL基因异常的AML中FAB分型以M5、M4居多,基因异常类型以MLL-PTD居多,伴MLL基因异常的ALL主要为B-ALL,基因异常类型主要是MLL-AF4。此类白血病对传统化疗反应不良,易复发,已成为疾病预后不良的标志,异基因移植有可能改善其预后。     

急性早幼粒细胞白血病的治疗进展和目前存在问题

急性早幼粒细胞白血病(APL)占急性髓系白血病(AML)的10%~15%,形态学上属于FAB(法国、美国和英国)分型中的AML-M3亚型,98%的APL具有t(15;17)(q22;q21)染色体异位和早幼粒细胞白血病-维A酸受体α(PML-RARA)融合基因[1]。因APL具有较高的出血倾向和早期死亡率,一度被认为是最凶险的急性白血病。1973年,Bernard     

骨髓增生异常综合征FAB和WHO分型的临床评价

目的:评价和分析骨髓增生异常综合征(MDS)从FAB分型到WHO分型的发展和临床意义。方法:对MDS患者分别用FAB分型及WHO分型进行分型,并对形态学、临床、实验室检查及预后资料进行对比分析。结果:MDS和急性髓性白血病(AML)均可出现病态造血。FAB分型中难治性贫血(RA)、原始细胞过多难治性贫血(RAEB)、转化中的原始细胞过多难治性贫血(RAEB-T)及AML之间生存率差异有统计学意义。WHO分型中RA与难治性血细胞减少伴多系增生异常(RCMD)之间生存率差异无统计学意义,RA与RAEB、RCMD与RAEB之间生存率差异有统计学意义,RAEB-Ⅰ与RAEB-Ⅱ之间生存率有显著差异。结论:WHO分型将FAB分型中的RA分为RA和RCMD并未显示出临床优越性。RAEB-T生存期比AML更短,因而将RAEB- T归为急性白血病,对临床治疗有好处。WHO分型按照原始细胞百分比将RAEB分为RAEB-Ⅰ和RAEB-Ⅱ,对临床诊断、治疗和预后有益。     

急性白血病bcl-2、bax测定的临床意义

目的探讨急性白血病(AL)患者bcl-2、bax的表达与临床治疗及预后的关系。方法采用S-P免疫组化法检测AL患者骨髓单个核细胞(BMMNC)上bcl-2及bax的表达水平,行AL疗效分析。结果急性髓细胞白血病(AML)和急性淋巴细胞白血病(ALL)患者bcl-2表达分别为(46.12±27.29)%和(57.94±32.19)%,均明显高于正常对照组(15.4±5.64)%(P<0.01),AML与ALL间bcl-2表达差异无统计学意义(P>0.05);49例AL临床治疗有效组bcl-2表达为(36.81±30.35)%,无效组为(56.36±26.78)%,无效组高于有效组(P<0.05)。各组间bax差异均无统计学意义(P>0.05)。结论bcl-2水平对判断AL疗效和预后有重要价值。     

CD117和CD34在急性白血病中的临床意义

目的:观察细胞表面分化抗原CD117和CD34在急性白血病中的表达及意义。方法:采用三色流式细胞术,CD45/SSC双参数散点图设门方法,对34例急性白血病患者[急性淋巴细胞白血病(ALL)9例和急性髓细胞白血病(AML)25例]骨髓白血病细胞的免疫分型及CD117和CD34的表达进行检测。结果:CD117在ALL和AML中表达率分别为0和76.0%(19/25),差异有统计学意义(P=0.000)。CD34在ALL和AML中表达率分别为77.8%(7/9)和60.0%(15/25),差异无统计学意义(P=0.815)。在AML-M3亚型中,CD34表达率为16.7%(1/6),显著低于非M3亚型73.7%(14/19)(P=0.007)。结论:CD117检测有助于ALL与AML的鉴别诊断,CD34检测有助于区分AML-M3亚型和非M3亚型。     

实时荧光定量PCR检测急性白血病的肝细胞生长因子mRNA的表达及其意义

目的 定量检测急性白血病(AL)骨髓单个核细胞(MNCs)的肝细胞生长因子(HGF)mRNA的表达,并探讨其临床意义.方法 制备67例初治AL患者的骨髓MNCs,提取定量MNCs的总RNA,并制备cDNA.应用实时荧光定量-PCR(FQ-PCR)检测HGF mRNA在AL的表达.结果 HGF mRNA在AL组的表达显著高于对照组(6.936 4±1.613,0.407±0.170,P<0.001),而急性髓系白血病(AML)和急性淋巴细胞白血病(ALL)的表达差异无统计学意义(7.127±1.911,6.635±0.934,P>0.05).在AL中,M5呈显著强表达(9.998±1.454),高于M2、M3、M4、L1、L2和L3表达(P<0.001);而后者间表达差异无统计学意义(P>0.05).HGF mRNA在性别、年龄间的表达差异也无统计学意义(P>0.05).缓解组HGF mRNA的表达明显低于未缓解组(6.393±1.165,8.041±1.848.P<0.005).结论 HGF mRNA在AL组与对照组、AL亚型间的表达存在差异,但在AML与ALL、AL患者的性别、年龄间的表达无差异,且低表达者疗效较好,提示HGF mRNA可能是AL诊治的又一理想指标.     

Flowcytometric Immunophenotypic Profile of Acute Leukemia: Mansoura Experience

Acute leukemia (AL) displays characteristic patterns of antigen expression, which facilitate their identification and proper classification. The purpose of this study is to evaluate the diagnostic usefulness of commonly used immune-markers for immunophenotyping of AL and to define the best immune-markers to be used for proper diagnosis and classification of AL. Besides, to recognize the frequency of different AL subtypes and the antigen expression profile in our Egyptian patients. We retrospectively analyzed the immunophenotypic data of 164 de novo AL patients from our institution during 2009 and 2010. Among these patients, 68.9% were classified as acute myeloblastic leukemia (AML) while 31.1% classified as acute lymphoblastic leukemia (ALL). The commonest FAB subtype in AML group was AML-M4/5 (34.5%) which may differ from most published data. As regard ALL, there were 74.5% with B-ALL and 25.5% with T-ALL. It was found that combined use of HLADR and CD34 was much more helpful in distinguishing APL from non-APL AML than either of these antigens alone. It was found that cCD79a and CD19 were the most sensitive marker for B-ALL while cCD3, CD7 and CD5 were the most sensitive antigens for T-ALL. Our analysis of AL phenotypes proved that employed antibody panels are adequate for proper diagnosis and classification of AL. Flowcytometry was found to be especially useful in the identification of AML-M0 and differentiation of APL from non-APL AML. Immunophenotyping results and FAB classification of our AL patients were comparable to internationally published studies apart from predominance of AML-M4/5 and more frequent APL.     

老年急性白血病患者50例临床特征分析

目的:探讨老年急性白血病(AL)的临床特点,以利于有效治疗。方法:回顾分析50例60岁以上的老年AL患者的临床资料,包括年龄分布、基础疾病、主要症状、临床特征、骨髓(BM)象、染色体、免疫分型、化疗的完全缓解(CR)率、BM抑制程度、病程及病死率。并与同期住院的66例中青年患者进行比较。结果:老年AL发病率占同期成人AL的27%(50/185)。AML的CR率35.7%(15/42),ALL的CR率33%(2/6)。其基础疾病的发病率86%、MDS转化为AML占20%、病程(35.5±16.5)d、BM抑制时间(18.5±6.5)d、病死率20%、染色体核型为-5、-7、+8、+21,以上均高于同期的中青年组(均为P<0.01)。结论:老年AL患者并存基础疾病和染色体核型异常是其病死率高的主要原因,目前尚无早期诊断和有效治疗老年AL的满意策略,呼吁重视此方面的研究。     

Classifying acute leukemia by immunophenotyping: a combined FAB-immunologic classification of AML

A panel of commercially available monoclonal antibodies and five heteroantisera were used to distinguish and subtype 138 cases of acute leukemia (AL). The immunophenotype was compared with the French-American-British (FAB) classification obtained on the cases. The immunophenotype discriminated acute myelogenous leukemia (AML) from acute lymphoblastic leukemia (ALL) and recognized cases not distinguished by cytochemistry (22% of cases), mixed lineage phenotypes (13% of cases), and cases with separate populations of lymphoblasts and myeloblasts (one case). Using the immunologic panel and derived criteria to subtype AML, correspondence of the immunophenotype to the FAB subtypes M1, M2, M4, and M5 was possible in greater than 80% of cases. A combined classification of the immunophenotype and FAB morphology/cytochemistry was devised for AML subtyping. It is recommended that immunophenotyping should be done at least in all cases with negative or inconclusive cytochemistry. At present, we suggest that until a "gold standard" for identifying leukemic subtypes is developed, the best method for typing acute leukemia is by using a combination of morphology, cytochemistry and immunophenotyping.     

P-glycoprotein quantitation in acute leukemia

Multi drug resistance(MDR) is a major problem in the treatment ofcancer and hematological malignancies. This resistance is multi factorial and is the result of decreased intra cellular drug accumulation. This is partly due to the presence of a 170KD intra membranous protein termed P-glycoprotein(P-gp) that is an energy- dependent efflux pump which has increased expression on drug-resistance cells. In this study we identified the presence of P-gp by staining with Fluorescent Iso Thio Cyanate (FITC) conjugated anti P-gp in acute leukemia patients and flow cytometry in addition to performing immunophenotype analysis and French, American British (FAB) classification. Results revealed that one fifth of leukemic patients expressed P-gp and this phenotype was more prevalent in Acute Undifferentiated Leukemia(AUL) and Acute Myelogenous Leukemia (AML) than in Acute Lymphoblastic Leukemia(ALL). Other findings showed a logical relationship between this phenotype and age groups. There was not any association between P-gp+ phenotype and FAB and Immunophenotyping sub classification, but there was a linear relationship between CD34 and CD7 expression and P-gp+ phenotype. The accumulation of P-gp molecule that was stated as Mean Fluorescence Intensity (MFI) on the blasts' membrane of AUL and AML patients showed marked increase in comparison to ALL. Furthermore MFI in P-gp+ relapsed patients was much more than P-gp+ pretreatment patients. Kepvords: Leukemia, Drug resistance, P-glycoprotein, Flowcytometry, FAB classification, Immunophenotyping, Mean Fluorescence Intensity.     

Leukemic cells and aberrant phenotypes in acute leukemia patients: a flow cytometry analysis

The aim of this study was to evaluate the heterogeneity of immunophenotype features in acute leukemia patients and to detect the presence of leukemia-associated immunophenotypes. We prospectively investigated the phenotype of blast cells from 44 adult acute leukemia patients using a large panel of monoclonal antibodies by multiparametric flow cytometry. Thirty-three patients were classified as AML according to the FAB classification. Eleven patients were diagnosed as ALL (10 cases B-ALL, 1 case T-ALL) according to both FAB and immunnophenotyping. We found leukemia-associated phenotypes in 28 of 33 AML patients (84.8%) and in 8 of 11 ALL patients (72.7%). In 61.1% of patients more than one aberrant phenotype was observed. Linear infidelity was the most frequent aberrancy in both AML (64.3%) and ALL (37.5%) subgroups. The present study shows that MFC is a helpful method for sufficient identification of leukemic cells and for determination of blast cells immunophenotype heterogeneity. The double stain flow cytometry in our study revealed aberrant phenotypes in up to 81.8% patients.     

Submicroscopic deletions at 7q region are associated with recurrent chromosome abnormalities in acute leukemia

BACKGROUND AND OBJECTIVES: Loss of heterozygosity (LOH) on the long arm of chromosome 7 (7q) has been frequently reported in several types of human cancer including hematologic malignancies. Moreover, monosomy of chromosome 7 and 7q deletions have been associated in acute myeloid leukemia (AML) with aggressive disease and poor prognosis. DESIGN AND METHODS: Using a panel of 11 polymorphic microsatellite markers at bands 7q21-q36, we investigated fifty patients (acute myeloid leukemia [AML], n=33 and acute lymphoid leukemia [ALL], n=17) for LOH, a hallmark of possible involvement of tumor suppressor genes. In parallel, the same acute leukemia (AL) cases were studied by conventional cytogenetics. RESULTS: A total of 48 spots of allelic loss were observed in 16 (32%) out of 50 patients (AML, n=11 and ALL, n=5). Among LOH+ve cases 3 showed chromosome 7 monosomies, whereas no cytogenetically detectable abnormalities were observed in chromosome 7 in the remaining 13. INTERPRETATION AND CONCLUSIONS: Comparison with karyotypic results indicated that presence of LOH at 7q21-q36 was significantly associated with other chromosomal aberrations. In fact, an altered karyotype was detectable in 87% of LOH+ve and in 52% of LOH(-ve) AL cases (p=0.024). In addition, LOH at 7q was prevalently associated with unfavorable cytogenetic lesions (p=0.013). Our study represents the first report of a significant association between LOH and recurrent chromosomal abnormalities in AL patients suggesting that the 7q21-q36, region may be an unstable area prone to chromosome breakage in patients with an abnormal karyotype.     

A single-center population-based consecutive series of 1500 cytogenetically investigated adult hematological malignancies: karyotypic features in relation to morphology, age and gender

During the 18-yr period 1976-93, a population-based series of 1586 adults with suspected or confirmed hematological malignancies were successfully cytogenetically investigated at a single center. Eighty-six cases were excluded due to unretrievable medical records or if analyzed only in remission or at relapse. The remaining 1500 medical records were reviewed regarding morphology and clinical parameters in order to investigate possible associations between karyotypic pattern (normal, 1, 2 or complex anomalies; specific abnormalities) and gender, age and morphological subgroups. The impact of time-period, i.e. 1976-87 vs. 1988-93, and referring center on cytogenetic findings was also studied. A total of 372 acute myeloid leukemias (AML), 389 myelodysplastic syndromes (MDS), 64 acute lymphoblastic leukemias (ALL) and 262 chronic myeloid leukemias (CML) were identified, altogether 1087 cases. Patients with other (n=261) or no hematological malignancies (n = 152) were excluded from the present analysis. Cytogenetic abnormalities were detected in 52% AML, 51 % MDS, 68% ALL and 97% CML, frequencies that did not differ significantly between the 2 time periods or referring centers. No significant age- or gender-related differences in karyotypic patterns were discerned in AML, MDS, ALL or CML, whereas the karyotypic patterns varied among the FAB groups in both AML (p= 0.001) and MDS (p < 0.001). The specific abnormalities t(8;21), t(15;17) and inv(16) were more common (p < 0.001) in younger AML patients and 5q- was more frequent in females with MDS (p<0.001). These findings indicate, in contrast to previous series, that neoplasia-associated karyotypic aberrations are not more common among older patients or in males.     

Clinical features of adult acute leukemia with 11q23 abnormalities in Japan: a co-operative multicenter study

To clarify the clinical features of adult patients with acute leukemia (AL) with 11q23 abnormalities, we performed a retrospective analysis of data from 58 adult Japanese patients: 51 with acute myeloid leukemia (AML), and 7 with acute lymphoblastic leukemia (ALL). The incidences according to fusion partners in AML were: t(9;11), 31.3%; t(11;19), 27.4%; t(6;11), 21.5%. The incidence of patients with t(11;19) was higher than those in the US and Europe, and the incidence of t(4;11) was lower than that in childhood. The results indicated the poor prognosis of AML with 11q23 abnormalities regardless of the fusion partners. AML patients with 11q23 aged <60 years in the first CR who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) showed a more favorable outcome than those treated without allo-HSCT, although the differences were not statistically significant (P = 0.322 for DFS, P = 0.138 for OS). This result suggests that treatment strategies including allo-HSCT may be considered in the first CR in cases of AML with 11q23 abnormalities. However, further studies involving a large number of cases are required to assess the effect of allo-HSCT on adult AL with 11q23 abnormalities.     

MLD according to the WHO classification in AML has no correlation with age and no independent prognostic relevance as analyzed in 1766 patients

Between February 1996 and December 2004, the German Leukemia Study Initiative registered 1766 consecutive patients for the acute myeloid leukemia (AML) 96 study, all of whom were diagnosed by central cytomorphology according to the French-American-British (FAB) and the new World Health Organization (WHO) classification. We focused our analysis on the prognostic impact of multilineage dysplasia (MLD) as a new parameter of the WHO classification for AML. We could not confirm the WHO statement that MLD occurs most frequently in older individuals, but we confirmed that MLD is often associated with an unfavorable cytogenetic profile (P < .001). In 1332 individuals receiving intensive AML therapy presence of MLD was negatively correlated with complete remission (P = .001) in univariate, but not in multivariate, analysis. Multivariate analysis of either event-free or overall survival again failed to show an independent prognostic significance of MLD besides age, cytogenetics, and, in part, NPM1/FLT3-ITD mutations. Our data support a reassessment of the WHO classification in the light of a more biologic understanding of AML. This study is registered at www.ClinicalTrials.gov as #NCT00180115.     

Flow cytometric analysis of cell-surface and intracellular antigens in the diagnosis of acute leukemia

To evaluate the usefulness of flow cytometric detection of intracellular antigens (Ags) in establishing proper lineage affiliation and its contribution to the diagnosis of acute leukemia, we studied 100 consecutive patients in whom acute leukemia was diagnosed between January 1997 and July 1998. Immunological classification was assessed using a three-line panel of monoclonal antibodies for phenotypic characterization of leukemic blast cells as proposed at the First Latin American Consensus Conference for Flow Cytometric Immunophenotyping of Leukemia. We found 74 cases of B-cell lineage acute lymphoblastic leukemia (ALL), seven cases of T-cell ALL, and 19 cases of acute myeloid leukemia (AML). In this study cytoplasmic (cy) CD79a, cyCD22, cyCD3, and cyMPO were highly sensitive, specific B, T, and myeloid markers that were expressed in virtually all cases of B and T cell ALL and in all subtypes of AML. Applied in combination with immunophenotyping this knowledge led to improvement in diagnostic precision and refinement of immunological classification, ensuring the selection of the most appropriate therapy for the patients studied. In conclusion, intracellular Ags detection was of utmost importance in establishing correct lineage affiliation in cases lacking expression of B, T, or myeloid surface Ags or disclosing equivocal or ambiguous immunophenotypic features and in identifying biphenotypic acute leukemia. In combination with FAB morphology and immunophenotyping, we were able to reliably classify all patients with acute leukemia in this study.