CD38 ZAP-70在B慢性淋巴细胞白血病中表达及其临床意义的研究术

目的：检测CD38及zeta链相关蛋白-70（zeta-associated protein-70,ZAP-70）在B慢性淋巴细胞白血病（B—CLL）中的表达,探讨其与临床的相关性。方法：采用流式细胞仪检测51例B慢性淋巴细胞白血病（B—CLL）患者骨髓或外周血白血病细胞CD38和ZAP-70的表达,结合其与患者临床分期、病情进展、化疗疗效进行分析。结果：1）44例患者中10例（22．73％）CD38和ZAP-70同时表达阳性,22例（50％）CD38和ZAP-70同时表达阴性。2）31例CD38^-患者中,初诊时处于Binet B＋C期者9例（29．03％）;11例CD38^＋患者中,初诊时处于Binet B＋C期者7例（63．64％）。CD38阳性、阴性表达在Binet分期上无显著差异（P〉0．05）。27例ZAP-70^-患者中,初诊时处于Binet B＋C期者6例（占22.22％）;16例ZAP-70^＋患者中,初诊时处于Binet B＋C期者10例（62．5％）。ZAP-70阳性、阴性表达在Binet分期上有显著差异（P〈0．05）。3）23例患者予氟达拉滨单药或联合CTX治疗,16例CD38^-表达者中,化疗有效（CR＋PR）者11例（68．75％）;7例CD38^＋表达者中,化疗有效者2例（14.29％）。11例ZAP-70^-表达者中,化疗有效者9例（81．82％）;9例ZAP-70^＋表达者中,化疗有效者1例（11．11％）。CD38和ZAP-70阳性、阴性的表达与化疗疗效有显著差异（P〈0．05）。结论：1）B—CLL患者CD38的表达和ZAP-70的表达有关。2）B—CLL患者ZAP-70阳性、阴性的表达与Binet分期有关系,ZAP-70的阳性表达患者较阴性表达者的病情进展快。3）B—CLL患者CD38和ZAP-70的阳性表达者较阴性表达者的化疗疗效差,预后欠佳。采用流式细胞术检测ZAP-70的表达结合CD38可以作为慢性淋巴白血病的临床分型和预后判断的一个参考指标。     

CD38 ZAP-70在B慢性淋巴细胞白血病中表达及其临床意义的研究

目的:检测CD38及zeta链相关蛋白-70(zeta-associated protein-70,ZAP-70)在B慢性淋巴细胞白血病(B-CLL)中的表达,探讨其与临床的相关性.方法:采用流式细胞仪检测51例B慢性淋巴细胞白血病(B-CLL)患者骨髓或外周血白血病细胞CD38和ZAP-70的表达,结合其与患者临床分期、病情进展、化疗疗效进行分析.结果:1)44例患者中10例(22.73%)CD38和ZAP-70同时表达阳性,22例(50%)CD38和ZAP-70同时表达阴性.2)31例CD38-患者中,初诊时处于Binet B+C期者9例(29.03%);11例CD38+患者中,初诊时处于Binet B+C期者7例(63.64%).CD38阳性、阴性表达在Binet分期上无显著差异(P>0.05).27例ZAP-70-患者中,初诊时处于BinetB+C期者6例(占22.22%);16例ZAP-70+患者中,初诊时处于Binet B+C期者10例(62.5%).ZAP-70阳性、阴性表达在Binet分期上有显著差异(P<0.05).3)23例患者予氟达拉滨单药或联合CTX治疗,16例CD38-表达者中,化疗有效(CR+PR)者11例(68.75%);7例CD38+表达者中,化疗有效者2例(14.29%).11例ZAP-70-表达者中,化疗有效者9例(81.82%);9例ZAP-70+表达者中,化疗有效者1例(11.11%).CD38和ZAP-70阳性、阴性的表达与化疗疗效有显著差异(P<0.05).结论:1)B-CLL患者CD38的表达和ZAP-70的表达有关.2)B-CLL患者ZAP-70阳性、阴性的表达与Binet分期有关系,ZAP-70的阳性表达患者较阴性表达者的病情进展快.3)B-CLL患者CD38和ZAP-70的阳性表达者较阴性表达者的化疗疗效差,预后欠佳.采用流式细胞术检测ZAP-70的表达结合CD38可以作为慢性淋巴白血病的临床分型和预后判断的一个参考指标.     

FC与FCR方案治疗慢性淋巴细胞白血病的临床回顾性分析

  目的  比较FC（氟达拉滨和环磷酰胺）和FCR（氟达拉滨、环磷酰胺和利妥昔单抗）方案治疗慢性淋巴细胞白血病（CLL）的临床疗效和预后影响。  方法  回顾性分析本院2002年12月至2012年1月应用FC或FCR方案治疗的58例CLL患者病例资料,比较两种方案的疗效和预后影响。  结果  FC组31例,FCR组27例。FCR组完全缓解（CR）率和总反应率（ORR）均高于FC组（44.4% vs. 19.4%,P=0.039;81.5% vs. 51.6%,P=0.017）。疗程结束后,FCR组患者获得微小残留病（microscopice residual disease,MRD）阴性比例高于FC组（37.0% vs. 12.9%,P=0.032）。FCR和FC组患者中高危遗传学亚组患者PFS较非高危遗传学亚组患者短（P值分别为0.011,0.027）,OS时间无明显差别。  结论  FCR方案是治疗CLL患者的更为有效的方案。      

胃癌根治术后切缘阳性患者的临床病理特征和预后分析

  目的  探讨胃癌根治术后切缘阳性患者的临床病理特征及其对预后影响。  方法  回顾性分析河北医科大学第四医院2011年1月至2016年1月收治的胃癌根治术后切缘阳性患者的临床病理资料。按1∶2随机数法选取同期收治的切缘阴性患者,比较阳性和阴性切缘患者的一般临床病理学特征及预后情况。  结果  共纳入73例切缘阳性患者,与同期纳入的146例切缘阴性病例比较,阳性组的肿瘤直径更大、更多位于贲门或全胃,组织学类型更差、Lauren分型趋于弥漫型、Borrmann分型多为Ⅲ～Ⅳ型、肿瘤浸润深度以T4a～4b为主、pTNM分期更晚,脉管浸润率及淋巴结转移率也更高,同时术者经验、手术方式的差异也与阳性切缘发生有关（均P<0.05）。全组共有205例患者获得完整随访,两组患者5年总生存（overall survival,OS）率及无进展生存（progression-free survival,PFS）率均有显著性差异（23.19% vs. 58.82%,15.94% vs. 47.06%,均P<0.001）。Cox多因素分析显示,切缘状态（P=0.012）、pTNM分期（P=0.023）及术后综合治疗（化疗/化疗联合放疗）（P<0.001）是影响胃癌预后的独立因素。  结论  胃癌根治术后切缘状态与多种临床病理特征相关,切缘阳性患者预后较差。      

循环肿瘤细胞与乳腺癌患者预后相关性的Meta分析

  目的  评估循环肿瘤细胞（circulating tumor cell,CTC）在预测乳腺癌患者预后中的作用。  方法  检索Medline、Embase、中国数字化期刊全文数据库（CNKI）、万方数据库及维普全文网中国内外相关文献,收集检测外周血肿瘤细胞在乳腺癌患者预后中的研究,使用Review Manager 5.1.4中的方差倒数法对无进展生存期（progression-free survival,PFS）和总生存期（overall survival,OS）进行Meta分析。  结果  共纳入33篇英文文献,总计5 393例乳腺癌患者。Meta分析结果提示CTC阳性组较CTC阴性组在PFS [HR=2.09（95%CI:1.72~2.55）,n=23,I2=57%]和OS[HR=2.49（95%CI:2.18~2.85）,n=24,I2=0]方面差异均有统计学意义（P < 0.01）。根据UICC（International Union Against Cancer）肿瘤分期进行亚组分析,结果显示CTC对不同期别肿瘤的PFS和OS也均有预测价值（P < 0.01）。  结论  外周血CTC阳性的乳腺癌患者与CTC阴性患者相比预后较差,CTC可以作为乳腺癌患者的预后预测指标。      

荧光原位杂交技术在检测慢性淋巴细胞白血病ATM基因缺失中的应用

 目的　探讨慢性淋巴细胞白血病（CLL）患者中ATM基因缺失情况及其与临床分期的相关性。方法　以骨髓涂片档案片为标本,运用荧光原位杂交（FISH）技术和SpectrumOrangeTM标记的位于11q22.3的序列特异性DNA探针ATM对28例初诊的CLL患者进行了ATM缺失的检测,临床分期按照Binet分期方法,采用Fisher确切概率法分析ATM基因缺失与患者初诊时临床特征间的关系。结果　FISH分析中,28例CLL患者中有4例（14.3 %）检出ATM基因缺失,9例Binet A期患者中1例（11.1 %） 存在del（ATM）异常;8例Binet B期患者中1例（12.5 %） 存在异常;11例Binet C期患者中2例（18.2 %） 存在异常。Fisher确切概率法分析显示ATM基因缺失与年龄、性别、临床分期无明显相关性（P＞0.05）。结论　以骨髓涂片档案片为样本进行FISH检测,方法简便,结果可靠,便于血液系统克隆性疾病的回顾性遗传学分析,ATM基因缺失是CLL患者常见的细胞遗传学改变,其对中国CLL患者的预后预测价值有待进一步深入研究。     

CD38及ZETA相关蛋白在慢性淋巴细胞白血病中的表达及其临床意义

 【摘要】　目的　探讨CD38及ZETA相关蛋白（ZAP-70）在慢性淋巴细胞白血病（CLL）患者中的表达及其与临床分期的关系,分析其对预后的影响。方法　采用流式细胞术检测42例CLL患者CD38及ZAP-70的表达水平,根据Rai分期将患者分为高危组（Ⅲ、Ⅳ期）（25例）和低中危组（0、Ⅰ、Ⅱ期）（17例）,了解CD38和ZAP-70表达在Rai分期的分布情况,结合患者病情进展、化疗疗效进行分析。结果　CLL患者中CD+38者占47.6 %（20／42）,其中高危组 64.0 %（16／25）,低中危组 23.5 %（4／17）,两组差异有统计学意义（χ2＝6.645,P＝0.014）; ZAP-70+者占40.5 %（17／42）,其中高危组 60.0 %（15／25）,低中危组 11.8 %（2／17）,两组差异有统计学意义（χ2＝9.772,P＝0.003）; ZAP-70+ CD+38和ZAP-70+ CD-38 患者多分布在高危组中,而ZAP-70- CD-38患者多分布在低中危组中,差异均有统计学意义（χ2＝10.076、9.346、6.005,均P＜0.05）。随访48个月（1～136个月）后 ZAP-70+ CD+38及ZAP-70- CD-38患者无进展生存期分别为19.0和58.0个月,差异有统计学意义 （χ2＝11.488,P＝0.003）,而ZAP-70+ CD-38和ZAP-70- CD+38 患者无进展生存时间分别为43.5和51.7个月,差异无统计学意义（χ2＝0.075,P＝0.784）。结论　CD38和ZAP-70在CLL病程后期（Rai分期Ⅲ期和Ⅳ期）表达高于病程早期（Rai分期0期、Ⅰ期和Ⅱ期）,CD38和ZAP-70均阳性CLL患者较单阳性患者病情进展快,生存期明显缩短。     

慢性淋巴细胞白血病外周血T细胞亚群及免疫球蛋白研究

 目的　分析慢性淋巴细胞白血病（CLL）患者的免疫状态及其与疾病分期的关系。方法　应用流式细胞术（FCM）分析60例B-CLL患者外周血T淋巴细胞亚群的变化，用免疫速率比浊法分析其中37例患者血清中IgG、IgA和IgM的变化，比较各Binet分期患者免疫功能的变化。结果　与正常对照组相比，60例B-CLL患者外周血中CD+3、CD+4和CD+8细胞比率均明显降低（P＜0.01），Binet B、C期组患者CD+4／CD+8比值也明显低于Binet A和正常对照组（P＜0.01）。37例B-CLL患者中有21例患者（56.8 %）出现有免疫球蛋白（Ig）减低的情况。Binet C期组Ig减低发生率高于Binet A和B期（P＜0.05），Binet C期组IgG、IgA、IgM的平均水平明显低于Binet A和B期组（P＜0.05）。结论　CLL患者存在细胞免疫和体液免疫功能的低下，与CLL病程密切相关。检测CLL患者外周血T细胞亚群及Ig水平的变化，对了解CLL免疫功能状态，从而判断病情具有重要作用。     

ZAP-70及CD38在慢性B淋巴细胞白血病中表达的意义

 目的　检测zeta相关蛋白（ZAP-70）及CD38在慢性B淋巴细胞白血病（B-CLL）中的表达,分析两者的相关性,并探讨其临床意义。方法　采用流式细胞仪检测21例B-CLL患者骨髓或外周血中ZAP-70及CD38的表达水平。结果　在21例B-CLL中9例ZAP-70阳性,8例CD38阳性。二者同时阳性8例,二者同时阴性8例。结论　ZAP-70及CD38可以为B-CLL提供重要的预后信息,阳性提示预后不良,二者具有相关性。     

Super-ARMS法检测肺腺癌患者血浆循环肿瘤DNA表皮生长因子受体T790M基因突变的临床研究

  目的  探讨真实世界中Super-ARMS法检测肺腺癌患者外周血标本循环肿瘤脱氧核糖核酸（circulating tumor DNA,ctDNA）中表皮生长因子受体（epidermal growth factor receptor,EGFR）T790M基因突变的临床应用价值。  方法  收集2019年1月至2020年6月在首都医科大学附属北京胸科医院确诊的肺腺癌患者307例,突变扩增系统（amplification refractory mutation system, ARMS）检测组织中EGFR基因突变情况,Super-ARMS法检测血浆中EGFR的基因突变情况。通过生存分析比较不同标本检测EGFR T790M基因突变患者的无进展生存时间（progression-free survival,PFS）。  结果  153例患者疾病进展接受再活检。74例进行组织再活检,其中34例（45.9%）检测到EGFR T790M基因突变。141例患者进行液体再活检,其中51例（36.2%）EGFR T790M基因突变。Kaplan-Meier生存分析显示,组织和外周血EGFR T790M突变阳性患者接受第三代EGFR酪氨酸激酶抑制剂（EGFR-tyrosine kinase inhibitors,EGFR-TKIs）的中位PFS比较差异无统计学差异（16.3个月 vs. 11.4个月,x2=1.138,P>0.05）。组织和外周血EGFR T790M突变阴性患者未接受第三代EGFR-TKIs治疗的中位PFS比较差异无统计学意义（7.0个月 vs. 7.0个月,x2=0.470,P>0.05）。  结论  真实世界中Super-ARMS法检测外周血标本有望应用于检测EGFR T790M 基因突变情况,外周血标本可一定程度上补充组织标本检测EGFR T790M基因突变结果,预测患者对第三代EGFR-TKIs治疗的疗效。      

ZAP-70 immunoreactivity is a prognostic marker of disease progression in chronic lymphocytic leukemia

The expression of zeta-associated protein 70 (ZAP-70) in chronic lymphocytic leukemia (CLL) seems to correlate with the mutational status of the immunoglobulin heavy-chain variable-region genes, clinical course and patient prognosis. The aim was to determine the prognostic significance of the immunohistochemical expression of ZAP-70 protein in CLL by means of the long-term follow-up of 108 patients. This study identified 3 patterns of ZAP-70 immunoreactivity: negative (58 patients, 54%), weakly positive (20 patients, 18%) and strongly positive (30 patients, 28%). Overall, ZAP-70 immunoreactivity correlated with an abnormal karyotype ( p = 0.017), a lymphocyte doubling time (LDT) of <6 months ( p = 0.001) and <12 months ( p = 0.01), Rai II - IV and Binet B - C stage ( p = 0.013), the clinical need for chemotherapy ( p < 0.001) and the need for more than 1 chemotherapy line ( p < 0.001). Kaplan-Meier analysis demonstrated that ZAP-70 immunoreactivity closely correlated with a shorter LDT ( p < 0.0001) and time from diagnosis to initial therapy ( p = 0.0001). The same significance was retained when the patients were stratified into the ZAP-70 immunoreactivity groups ( p < 0.0001). This study shows that ZAP-70 immunoreactivity can be a reliable prognostic marker in CLL and proposes a system for evaluating the results. The observations support the inclusion of the immunohistochemical expression of ZAP-70 in clinical trials involving CLL patients.     

The prognostic evaluation of CLLU1 expression levels in 50 Chinese patients with chronic lymphocytic leukemia

Chronic lymphocytic leukemia (CLL) is characterized by the relentless accumulation of monoclonal B cells with the appearance of small mature lymphocytes and a characteristic CD5 and CD19 co-expression immunophenotype. The incidence of CLL is lower in Asian countries than in Western countries, where CLL is the most common leukemia. To investigate CLLU1 expression in CLL and explore the relationship between CLLU1 expression and alternative prognostic markers, we measured CLLU1 expression levels by semiquantitative RT-PCR in a cohort of 50 Chinese patients with CLL. Analyses of IgVH somatic mutational status, ZAP-70 expression, CD38 expression, and chromosomal aberrations were also performed. The expression of CLLU1 mRNA was determined in 26 of the 50 cases (52%), among which 7 at Binet A (7/21, 33.33%) and 19 at Binet B + C (19/29, 65.52%). The expression levels of CLLU1 were significantly increased in B + C CLL patients at Binet stage compared with those at Binet stage A (P = 0.005). Data for the IgVH somatic mutational status were available for 20 patients with known CLLU1 expression. Five (25%) patients, all expressed CLLU1 mRNA, displayed unmutated IgVH gene usage. While in 15 patients (15/20, 75%) with mutated IgVH gene, only 6 were CLLU1 positive. Patients with unmutated IgVH genes expressed higher levels of CLLU1 than did those with IgVH mutations (P < 0.05). Among 24 CD38(+)-CLL cases, 17 (70.83%) were CLLU1 positive, whereas only 9 (34.62%) positive cases were identified in 26 CD38(-)-CLL patients. Thus, the expression of CLLU1 in CD38(+)-CLL was significantly higher than that in CD38(-)-CLL. However, no significant difference of CLLU1 expression was found between ZAP-70(+) (14/22, 63.64%) and ZAP-70(-) (12/28, 42.86%) patients (P > 0.05). We conclude that CLLU1 expression was significantly associated with clinical stages, IgVH somatic mutational status and CD38 expression, and might be an important prognostic factor in CLL patients.     

ZAP-70 expression, as detected by immunohistochemistry on bone marrow biopsies from early-phase CLL patients, is a strong adverse prognostic factor.

Zeta-associated protein-70 (ZAP-70), mostly assessed by flow-cytometry (FC), recently emerged as reliable prognostic factor in chronic lymphocytic leukaemia (CLL) at presentation. We evaluated ZAP-70 expression in 156 CLL patients by immunohistochemistry (IHC) on formalin-fixed bone marrow (BM) biopsies at diagnosis. At presentation, 117 patients (75%) were with Binet stage A, 27 (17%) stage B and 12 (8%) stage C. Median follow-up was 61 months (range 6-242). ZAP-70 was expressed in neoplastic lymphocytes of 69 patients (44%). Concordance between ZAP-70 by IHC and ZAP-70 by FC, immunoglobulin heavy chain variable genes (IGHV) mutational status and CD38 expression was found in 41/46 (89%), 41/49 (80%) and in 60/88 (68%) tested cases, respectively. ZAP-70 expression significantly correlated with advanced Binet stage (B-C), diffuse BM infiltration, increased lactate dehydrogenase (LDH) and beta2-microglobulin serum levels and lymphocyte doubling time <12 months. ZAP-70 positivity was significantly related to poorer time to progression (median 16 months vs 158 of ZAP-70-negative cases) (P<0.0001) and overall survival (median 106 months vs not reached) (P=0.0002); this correlation was confirmed at multivariate analysis. ZAP-70 expression correlated with poorer outcome also when evaluated only in the 117 stage A patients. In conclusion, immunohistological detection of ZAP-70 on formalin-fixed BM biopsies at diagnosis appears a useful methodological approach to identify patients with poor prognosis in CLL.     

Expression of ZAP-70 protein correlates with disease stage in chronic lymphocytic leukemia and is associated with, but not generally restricted to, non-mutated Ig VH status

The mutational status of immunoglobulin variable region genes (Ig V_H) is a well established prognostic parameter in chronic lymphocytic leukemia (CLL). Recently, a subset of genes with a characteristic expression profile correlating with the mutational status of B-CLLs has been identified. One of the overexpressed genes in the prognostically unfavorable group of CLL patients with unmutated Ig V_H genes encodes for the protein tyrosine kinase ZAP-70, which is physiologically involved in T-cell signaling. Since ZAP-70 has been described to be prognostically relevant in CLL, we analyzed the possible relationship of its expression to the mutational status of Ig V_H genes as well as to other prognostic factors in CLL and indolent lymphomas. The mutational status of Ig V_H genes was analyzed by seminested PCR, direct sequencing and comparison with the sequences of the EMBL databases in 60 samples of patients with B-CLL and 18 samples of patients with indolent B-cell malignancies. ZAP-70 protein expression was assessed in all samples by immunoblotting and for semiquantitative analysis the ratio of ZAP-70 to tubulin expression was calculated. ZAP-70 protein was found to be expressed in all investigated B-cell malignancies. Expression levels varied within a wide range in each entity. The highest mean level of ZAP-70 expression was observed in unmutated B-CLLs, however, with broad expression variability. High levels of ZAP-70 expression correlated with higher stage Binet B or C and with unmutated Ig V_H genes. Overall survival rates estimated by Kaplan-Meier curves did not differ among patients with high or low ZAP-70 expression. We conclude that ZAP-70 is associated with the mutational status of Ig V_H genes, but this expression pattern is not present in all individual cases. Furthermore, high levels of ZAP-70 correlated with Binet stages B or C indicating an involvement of ZAP-70 in mechanisms promoting growth of B-CLL cells.     

Downregulated Dicer expression predicts poor prognosis in chronic lymphocytic leukemia

Chronic lymphocytic leukemia (CLL) is the most common leukemia in the western world. Alterations in microRNAs (miRNAs) expression have been proposed to play a role in CLL pathogenesis. Dicer and Drosha are the main regulators of miRNA biogenesis, and deregulation of their expression has been indicated as a possible cause of miRNA alterations observed in various cancers. To investigate the role of Dicer and Drosha in CLL, we assessed the expression of Dicer and Drosha and their correlation with other prognostic factors, including Binet stages, immunoglobulin heavy chain variable gene (IGHV) mutation status, TP53 mutation status, ZAP-70 protein and CD38 expression level in 165 CLL patients by using real-time polymerase chain reaction methods. Patients with unmutated IGHV genes had significantly lower expression of Dicer than patients with IGHV mutations. The lower expression level of Dicer was also significantly associated with higher level of CD38 and ZAP-70, and more aggressive Binet stage. We also analyzed Dicer expression in different cytogenetic subgroups. Lower Dicer level was found in patients with unfavorable cytogenetic aberrations (deletion in 17p13 or 11q22.3) in contrast to higher level in good risk cytogenetics (deletion in 13q14 as the sole abnormality). Furthermore, the lower expression of Dicer in CLL shows a strong association with shorter overall survival (OS) (P = 0.0046) as well as with reduced treatment free survival (TFS) (P = 0.0006). By contrast, no differences in the expression of Drosha among these groups of patients were observed. Our data suggest that Dicer expression may play an important role in the progression and prognosis of CLL.     

CD38 as a prognostic factor in Chinese patients with chronic lymphocytic leukaemia

B-cell chronic lymphocytic leukemia (CLL) is the most common type of adult leukemia in the Western countries, however, infrequent in the Eastern. It shows a remarkable heterogeneity, with some patients having an almost normal lifespan, others surviving only several years after diagnosis despite intensive therapy. To explore the prognostic significance of CD38 expression in Chinese patients with CLL, multi-parameter flow cytometry was used to detect the expression of CD38 on CD5(+)CD19(+) cells of 147 patients. CD38 expression and its association with some other prognostic factors such as Binet stage, lymphocyte count in peripheral blood, serum lactate dehydrogenase (LDH), beta2-microglobulin (beta2-MG), ZAP-70 expression and cytogenetic abnormalities were analyzed. The Kaplan-Meier method was used to construct survival curves, and results were compared using the log-rank test. Univariate and multivariate Cox regression analyses were used to assess associations between survival time and potential risk factors. Out of the 147 CLL patients, positive expression of CD38 was found in 45 (30.6%) cases. CD38-positivity identified a subgroup of CLL patients with aggressive disease of Binet stage at the time of the test (P=0.036). Furthermore, the presence of higher serum LDH and beta2-MG levels at diagnosis was strongly correlated with CD38-positive (P=0.016 and 0.025, respectively). Prognostically unfavorable cytogenetic abnormalities, including 17p13 and 11q22 deletions, were significantly more frequent in CD38-positive patients than in CD38-negative ones (P=0.047 and 0.001, respectively). There was no significant difference between CD38-positive and CD38-negative groups in molecular cytogenetic aberrations of del(6q23), del(13q14), 14q32 translocation, or trisomy 12. In addition, in CD38-positive patients, the percentage of leukemic cells expressing ZAP-70 protein was not significantly higher than in CD38-negative ones (P=0.120). CD38 expression was associated with poor outcome. Patients with positive expression of CD38 had significantly shorter overall survival (mean, 81 months) than patients without CD38 expression (mean, 179 months) (P=0.015). Univariate analysis showed that serum levels of LDH and beta2-MG, del(17p13) and CD38 expression were the significant factors in determining overall survival (OS). Del(17p13) and CD38 expression were the variables strongly associated with OS by multivariate Cox regression analysis. It was showed that the patients with high level of CD38 expression had poorer outcome; CD38 was a good predictor of OS in Chinese patients with CLL.     

Prognostic and Predictive Significance of Smudge Cell Percentage on Routine Blood Smear in Chronic Lymphocytic Leukemia

Introduction/BackgroundSmudge cells are ruptured lymphocytes present on routine blood smears of chronic lymphocytic leukemia (CLL) patients. We evaluated prognostic and predictive significance of smudge cell percentage on a blood smear in CLL patients.Materials and MethodsWe calculated smudge cell percentages (ratio of smudged to intact cells plus smudged lymphocytes) on archived blood smears of 222 untreated CLL patients registered at Institute Rotary Cancer Hospital, All India Institute of Medical Sciences, New Delhi over the past 12 years.ResultsThe male:female ratio was 3:1, and median age 60 (range, 28-90) years. Median absolute lymphocyte count was 42 × 10⁹/L. The median smudge cell percentage was 29.6% (range, 4%-79%). We found no correlation of proportion of smudge cells with age, sex, lymphocyte count, organomegaly, or response to therapy, although there was a significant correlation with the Rai stage at diagnosis. Median smudge cell percentage in stage 0 and I was 33% (range, 12%-79%), in stage II 31% (range, 12%-61%), and stage III and IV 21% (range, 4%-51%) (P < .001). Patients with ≤ 30% smudge cells had a shorter median progression-free period (PFP) of 30 months compared with patients who had more than 30% smudge cells (PFP, 45 months; P = .01). The 5-year survival rate was 51% for patients with 30% or fewer smudge cells, and it was 81% for patients with more than 30% smudge cells (P < .001) at a median follow-up of 3.5 years.ConclusionSimple and inexpensive detection of smudge cells on routine blood smears seems useful in predicting progression-free and overall survival in CLL patients and might be beneficial in countries with limited resources.     

Clinical features and outcome of Chinese patients with monoclonal B-cell lymphocytosis

B-cell chronic lymphocytic leukemia (CLL) is the most common type of adult leukemias in the Western countries, however, infrequent in the Eastern. A diagnosis of CLL requires a count of B-lymphocytes ≥5.0 × 10⁹/L. Asymptomatic person with <5.0 × 10⁹/L B-lymphocytes is defined as monoclonal B-cell lymphocytosis (MBL). To compare the clinical characteristics, prognostic factors, and outcome of Chinese patients with MBL and CLL, we present a study from our single centre of 20 patients with MBL and 136 patients with CLL. The factors included: age at diagnosis, gender, direct antiglobulin test (DAT), immunoglobulin heavy chain variable gene (IgHV) mutational status, ZAP-70 protein, CD38 expression level, and molecular cytogenetic aberrations were analyzed in MBL and CLL subgroups. The Kaplan–Meier method was used to construct survival curves, and results were compared using the log-rank test. Patients in the MBL category were slightly older than in the CLL category. There was no significant difference of these clinical and biological characteristics between patients in MBL subgroup and early stage CLL (Binet A). The incidence of positive DAT was significantly increased in CLL patients at Binet B and C, compared with MBL (P = 0.036). IgHV gene mutation in MBL is skewed, with more than 92.3% of subjects harbored mutated IgVH genes (P = 0.025). The proportion of MBL patients with a 13q14 deletion or trisomy 12 was similar to that of CLL patients. Moreover, markers associated with poor prognosis (deletion of 11q22 or 17p13) in these MBL populations were less than those in Binet B and C CLL patients (P = 0.025). No statistically significant differences in ZAP-70 and CD38 status were observed between the MBL and CLL subgroups. During a median follow-up period of 45.5 months, MBL patients had a low probability of progression, with no patients transformed to aggressive non-Hodgkin's lymphoma or dying of CLL-related causes. The overall survival of MBL was very similar to Binet A CLL, but longer than that of CLL patients at advanced stages (Binet B and C) (P = 0.024). Our study demonstrated that a more indolent clinical course and superior clinical outcome for patients with MBL compared to CLL.     

Multi-drug resistance mediated by P-glycoprotein overexpression is not correlated with ZAP-70/CD38 expression in B-cell chronic lymphocytic leukemia

ZAP-70 and CD38 expression can identify B-cell chronic lymphocytic leukemia with an inferior clinical outcome. Many groups have investigated the meaning of the expression of these two proteins and the correlation with the bad prognosis in B-CLL. But nobody has investigated the relation between the multidrug resistance mediated by Pgp overexpression (MDR1) and ZAP-70/CD38 coexpression. Forty-one untreated and stage A patients, either ZAP-70⁺CD38⁺ or ZAP-70^-CD38^-, were tested to determine the MDR1 status. MDR1 was observed in 41% of CLL ZAP-70⁺CD38⁺ and in 37% of CLL ZAP-70^-CD38^-. The difference was not significant (p = 0.745). Patients with ZAP-70 and CD38 positive CLL can not be candidates for MDR1 antagonists.