典型与不典型免疫表型慢性淋巴细胞白血病的预后相关因素分析

Objective To analyze the proguostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, abso-lute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations. Methods According to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophe-notype CLL, 16 with score 4 or 3 were atypical immunophenotype CI,L. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, slg, CD20, CD79h expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations. Results There was no significant difference be-tween the two groups in sex, age, ZAP-70 and IgVH mutation status (P =0.398, P =0. 189, P =0.268 and P =0. 131, respectively). The incidence of ALC≥50 × 109/L, Binet B + C, CD38 ≥30% in atypical CLL patients(43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P = 0. 026, P < 0. 01 and P = 0. 026, respectively). The proportion of typical patients (26. 8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P = 0. 022). Conclusion There were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.     

典型与不典型免疫表型慢性淋巴细胞白血病的预后相关因素分析

Objective To analyze the proguostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, abso-lute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations. Methods According to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophe-notype CLL, 16 with score 4 or 3 were atypical immunophenotype CI,L. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, slg, CD20, CD79h expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations. Results There was no significant difference be-tween the two groups in sex, age, ZAP-70 and IgVH mutation status (P =0.398, P =0. 189, P =0.268 and P =0. 131, respectively). The incidence of ALC≥50 × 109/L, Binet B + C, CD38 ≥30% in atypical CLL patients(43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P = 0. 026, P < 0. 01 and P = 0. 026, respectively). The proportion of typical patients (26. 8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P = 0. 022). Conclusion There were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.     

典型与不典型免疫表型慢性淋巴细胞白血病的预后相关因素分析

Objective To analyze the proguostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, abso-lute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations. Methods According to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophe-notype CLL, 16 with score 4 or 3 were atypical immunophenotype CI,L. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, slg, CD20, CD79h expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations. Results There was no significant difference be-tween the two groups in sex, age, ZAP-70 and IgVH mutation status (P =0.398, P =0. 189, P =0.268 and P =0. 131, respectively). The incidence of ALC≥50 × 109/L, Binet B + C, CD38 ≥30% in atypical CLL patients(43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P = 0. 026, P < 0. 01 and P = 0. 026, respectively). The proportion of typical patients (26. 8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P = 0. 022). Conclusion There were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.     

形态学酷似幼淋巴细胞白血病的急性髓系白血病

为了探讨形态学酷似幼淋巴细胞白血病（PLL）的急性髓系白血病（AML）的临床、骨髓细胞形态学、免疫表型及细胞遗传学特点，用显微镜观察骨髓细胞形态学特点，用流式细胞术检测骨髓细胞的免疫表型，用传统的细胞遗传学方法分析骨髓细胞核型，用荧光原位杂交技术测定骨髓细胞的杂交信号。结果表明：该患者的临床特点表现为急性白血病，骨髓细胞形态学提示为淋巴系统疾病，但免疫表型分析亦为髓系抗原表达，细胞遗传学分析该例患者存在8号染色体三体，对化疗反应性差。结论：急性髓系白血病细胞形态学可表现淋巴细胞样特点，结合免疫分型有助于进一步确定白血病细胞的类别分型，     

合并骨髓坏死的髓系抗原及Ph染色体阳性B急性淋巴细胞白血病一例

为加强对骨髓坏死（BMN）的病因及病理过程复杂性的认识，了解髓系抗原阳性（My^＋）及Ph染色体阳性（Ph^＋）的B急性淋巴细胞白血病（ALL）临床表现的多样性，报道分析了1例以BMN为首发症状的My^＋Ph^＋B-ALL并进行讨论。结果表明：该病例临床特征复杂而多样，通过骨髓涂片和活检，免疫分型，染色体核型分析及FISH明确了诊断。积极治疗原发病改善了患者的预后。结论：My^＋Ph^＋B-ALL并发BMN是一种罕见疾病，应采取多种方法检查明确诊断并积极治疗。     

ZAP-70在24例B细胞慢性淋巴细胞白血病中的表达研究

为了研究慢性淋巴细胞白血病中zeta链相关蛋白-70（zeta-associated protein-70，ZAP-70）的表达及其与其他预后因素的相关性，应用四色流式细胞术检测24例B细胞慢性淋巴细胞白血病（B-CLL）患者骨髓或外周血白血病细胞ZAP-70和CD38的表达。结果显示：①37．5％B-CLL患者表达ZAP-70，其中Binet A期患者有20％（3／15）表达ZAP-70^＋，Binet B＋C期患者有66．7％（6／9）表达ZAP-70^＋，ZAP-70^＋表达在BinetA期和Binet B＋C期之间有显著差异（P〈0．05）；②29．1％B-CLL患者表达CD38，其中Binet A期患者有3例；在此3例中2例同时伴有ZAP-70^＋，CD38^＋在Binet A期和Binet B＋C期之间无显著差异（P〉0．05）；③83．3％（20／24）患者表达ZAP-70^＋CD38^＋或ZAP-70^-CD38^-，ZAP-70和CD38存在相关性（P〈0．05）。结论：在常规实验室可以采用流式细胞术检测ZAP-70，ZAP-70高表达与B-CLL临床分期、染色体及CD38表达等预后相关因素有一定的相关性。     

骨髓间充质干细胞与K562细胞共培养对IL-8表达的影响

为了探讨骨髓间充质干细胞（mesenchymal stem cells,MSCs）与白血病细胞共培养后对血管生成因子IL-8表达的影响,本研究建立了MSC和K562细胞的共培养体系,包括直接接触和MSC条件培养液,应用RT-PCR方法检测MSC和K562细胞IL-8mRNA的表达。结果显示,与MSC直接接触和经MSC条件培养液作用后,K562细胞IL-8的表达均明显高于K562细胞单独培养组（p〈0.01）。与K562细胞共培养的MSC,其IL-8的表达亦明显高于单独培养的MSC（p〈0.01）。结论：MSC与K562细胞通过直接接触和某些细胞因子相互作用可促进血管生成因子IL-8的表达。     

母女同患慢性淋巴细胞白血病及其家系调查

慢性淋巴细胞白血病（CLL）是一种以小的、成熟淋巴细胞在外周血、骨髓以及淋巴组织聚集为特征的淋巴系统恶性增殖性疾病，且多为B细胞的克隆性增殖，即B细胞．CLL（B-eLL）。CLL是西方国家最常见的白血病类型，占全部白血病的三分之一。目前CLL病因尚不完全清楚，许多因素与CLL发病有关，包括物理因素、化学因素、生物因素和遗传因素，其中家族性遗传因素在西方国家已被公认。我国CLL发病率较低，占全部白血病的3％左右，母女同时患CLL未见报道，现将我科近期诊治的1对母女同患CLL的临床资料及家系调查报道如下。     

蛋白酶体抑制剂PS-341对多发性骨髓瘤骨髓间充质干细胞多种细胞因子表达的影响

为了探讨蛋白酶体抑制剂PS-341(bortezomib,velcade)对多发性骨髓瘤(MM)患者间充质干细胞(MSC)细胞因子分泌的影响,用含10%FBS的RPMI1640培养液培养11例MM患者MSC,细胞数达5-10×105时以终浓度50nmol/L的PS-341作用4小时,用逆转录聚合酶链反应(RT-PCR)检测IL-6、IL-1β、SCF细胞因子表达。结果表明:PS-341作用后MM病人MSC细胞因子的表达有显著下降,IL-6、IL-1β、SCF的表达与对照组比较,其显著性检验结果分别为P<0.05、P<0.01、P<0.05,其中IL-1β尤为明显;难治复发组与缓解组比较,IL-1β的表达有显著差异,完全缓解(CR)组IL-1β表达受抑制更明显,IL-6、SCF两组间表达无明显差异;PS-341治疗的1例患者MSC在PS-341作用后IL-1β未见表达;IL-1β的表达与否对IL-6、SCF的表达无影响。结论:蛋白酶体抑制剂PS-341下调MM患者MSC IL-6、IL-1β、SCF细胞因子的表达。     

人骨髓间充质干细胞对脐血干细胞体外扩增支持作用的研究

目的：探讨以人骨髓间充质干细胞(bone nlarrow-mesenchymal stem cells，BM-MSC)为基质的无血清培养方法，体外扩增脐血造血干细胞(cord blood stem cells，CBSC)，研究BM—MSC支持造血的功能，以能最终用于临床。方法：用含血小板生成素(TPO)、干细胞因子(SCF)、fit3／flk2配体(FL)和粒细胞-集落刺激因子(G—CSF)的无血清培养液，比较有或无MSC条件下，体外扩增脐血CD34^ 细胞两周后检测总细胞TC、CD34^ 细胞、集落形成单位(CFU)和长期培养-起始细胞(LTC—IC)增加倍数。结果：在TPO、FL、SCF、和G-CSF的作用下，经两周体外扩增后有MSC组的TC、CD34^ 细胞、CFU—GM、CFU—C和LTC—IC数较起始分别增加了427、39、125、104和16倍。单纯用上述4种细胞因子的无MSC组，TC、CD34^ 细胞、CFU—GM、CFU—C数分别增加了62、11、25、24倍，但LTC-IC只有扩增前的0．8倍。结论：以人BM—MSC为基质的无血清体外培养体系可以更有效扩增CBSC，有潜在的临床应用价值。