T淋巴细胞内mTOR/S6途径在难治/复发再生障碍性贫血发病机制中的作用

Objective To explore the activation status of signal pathway of mTOR/S6 in bone marrow(BM) T lymphocytes of refractory/relapsed aplastic anemia patients(AA),and the effects of rapamycin (RAPA)and CTLA-4 immunoglobulin(CTLA-4 Ig)on this pathway.Methods BM was collected from 13refractory/relapsed AA patients,8 newly diagnosed severe AA(SAA)patients and 10 iron deficiency anemia (IDA)(as controls)patients,and cocultured with RAPA and CTLA-4 Ig.The expression of p-mTOR,p-S6 and Interferon γ(IFN-γ)in CD3+ T cells was measured by flow cytometry(FCM).Results ①The expression of p-mTOR,P-S6 and IFN-γ in CD3+ T cells in refractory/relapsed AA group were significantly higher than those in controls(P＜0.01).②The expression of P-mTOR and p-S6 in T cels in newly diagnosed SAA group,Was similar to those in controls(P＞0.05),but significantly lower than those in refractory/relapsed AA group(P＜0.01).The expression level of IFN-γ in T cells were significantly higher than that in controls (P＜0.01).③On exposure to RAPA,The levels of p-mTOR,p-S6 and IFN-γ in T cells in refractory/relapsed AA patients were significantly lower than those before the exposure(all P＜0.05).And so were when exposed to CTLA-4 Ig(all P＜0.01).Conclusion ①The mTOR/S6 signal pathway is activated in refractory/relapsed AA.②The expression of p-mTOR,p-S6 and IFN-γ in refractory/relapsed AA Can be suppressed by RAPA or CTLA-4 Ig.③The signal pathway of CD28/mTOR/S6/IFN-γ might take part in immune pathogenesis of refractory/relapsed AA.     

一株新的急性髓系白血病细胞系SH-2的建立及其鉴定

Objective To establish and characterize a novel human myeloid leukemia cell line SH-2. Methods Bone marrow mononuclear cells(BMMNC) isolated from a AML-M2 patient, who failed to ob-tain complete remission after chemotherapy and allogenic bone marrow transplantation were passed in a long term IMDM culture medium supplemented with 20% fetal calf serum. Stromal cells were retained and rh-IL-3was added in the culture system. A new human myeloid leukemia cell line SH-2 was successfully established with a cytogeuetic characteristics of a loss of Y chromosome(- Y), a derivative chromosome 16 resulting from unbalanced translocation between chromosome 16 and 17, monosome 17, trisomy 19 and p53 alteration. Vari-ous methods were employed to characterize SH-2 cell line. Results SH-2 cells has been maintained without cytokine and stromal cells for more than 3 years without EB virus and mycoplasma contamination. SH-2 cells had the basically same morphological, immunophenotypic and cytogenetic features as the patient' s leukemia cells did, such as myeloid morphology, an immunophenotype of CD13+ , CD33+ , CD56+ , CD16/56+ and a hypodiploid karyotype of 45, X, - Y, der(16) t(16;17) (q24;q12) , - 17, + 19, which were gradually de-creased and replaced by the near-tetraploid cells with a karyotype of 73 - 102 (80), XX, - Y, - Y, del (lq31) ×2, der(16)t(16;17) (q24;q12) ×2, - 17, - 17, + 19, + 19. FISH and multiple FISH delineated all the abnormalities and revealed a loss of one p53 allele due to monosomy 17. DNA direct sequencing detec-ted a point mutation of CAG to CAT at codon 576 of exon 5 in another p53 allele. RT-PCR showed that SH-2 cells expressed apoptosis-related genes (bcl-2, Fas, GST- π and p21) rather than MDR-related genes. Short tandem repeat PCR provided powerful evidence for the derivation of SH-2 cell line from the patient' s leukemia cells. SH-2 cells had certain colony formation and tumorigenic capacities in nude and SCID mice. Conclu-sion SH-2 is a new myeloid leukemia cell line with a unique biology background, and will provide a useful tool for leukemia research.     

FCM和细胞遗传学检查在恶性淋巴瘤诊断中的意义

目的评价流式细胞术联合细胞遗传学检查在恶性淋巴瘤确诊中意义。方法笔者医院2009~2012年29例门诊及住院疑似淋巴瘤患者淋巴组织标本,制备单个核悬浮细胞,分别行流式细胞术检测细胞表面抗原的表达和R-显带检测细胞遗传学细胞核型。结果 29例患者中8例病理诊断为反应性增生或慢性炎,通过流式细胞仪和细胞遗传学诊断恶性淋巴瘤,12例患者发现核型异常,流式细胞术细胞表面抗原的检测可区分B淋系还是T淋系,区分不同亚型有指导意义,12例患者检测到异常核型,其中4例淋巴结病理提示淋巴结炎或者增生但核型发现异常染色体改变,肯定了为恶性改变,并发现有与亚型相关的特异性核型改变t(11;14),t(14;18),9例患者为复杂核型,提示预后不良。结论淋巴组织病理学检查是诊断恶性淋巴瘤的基础,结合流式细胞术以及染色体检查可以提高恶性淋巴瘤的诊断率,并对淋巴瘤亚型分型以及预后判断具有指导性意义。     

老年骨髓增生异常综合征患者铁过载的防治

目的 探讨老年骨髓增生异常综合征(MDS)患者铁过载的防治方法.方法 选取42例年龄≥60岁,病程≥3年的门诊MDS患者作为研究对象.采用常规治疗(雄激素、维甲酸、环孢素A等药物治疗、输注红细胞纠正贫血、抗感染、保肝等支持治疗),每周监测血细胞分析、生化、血清铁蛋白(SF),当SF≥1 000 μg/ml时,接受去铁胺祛铁治疗,随访3个月.结果 42例MDS患者检测出铁过载患者8例,血清铁蛋白为1 000～3 500μg/ml,铁过载患者接受常规治疗加去铁胺治疗,其余患者仍接受常规治疗.去铁胺治疗后1个月、2个月、3个月血清铁蛋白降低疗效显著(P＜0.05).结论 定期监测血清铁蛋白及去铁胺治疗对防治老年MDS患者铁过载具有重要的意义.     

50例慢性粒细胞白血病慢性期患者随访8年预后评价

目的 随访慢性粒细胞白血病(CML)患者,了解患者长期生存状况.方法 通过多种形式的健康教育、病友联谊会、心理护理、预约复诊、电话回访等综合干预,对选择不同治疗方法的50例慢性粒细胞白血病慢性期患者进行随访,定期监测血象、骨髓象、融合基因(BCR/ABL)、Ph染色体.结果 随访8年(2004年1月1日～2011年12月31日),长期生存470 (94％),其中生存期8～9年25例(50％),10～15年17例(34％),18年2例(4％),21～24年3例(6％),死亡3例(6％),无失访.随访结束47例长期生存患者骨髓象缓解44例(93.61％),融合基因(BCR/ABL)未检出42例(89.36％)、Ph染色体阴性45例(95.74％).结论 对选择不同治疗方法的CML慢性期患者进行综合干预,提高患者复诊率、延长生存期有明显效果.     

异基因造血干细胞移植治疗NPM1基因突变阳性骨髓增生异常综合征14例临床疗效北大核心CSCD

NPM1基因突变主要见于急性髓系白血病(AML),但也存在于骨髓增生异常综合征(MDS)和骨髓增殖性肿瘤(MPN)[1]。在MDS患者中,NPM1基因突变率约为4%[2-3]。NPM1基因突变阳性MDS患者往往具有原始细胞比例高、病程侵袭和易向AML转化的特点,预后分层多隶属中/高危组,具有异基因造血干细胞移植(allo-HSCT)适应证[1,4-5]。NPM1基因突变AML患者具有独立的预后特征[6]。本研究对近年来在本中心接受allo-HSCT治疗的14例NPM1基因突变阳性MDS患者进行回顾性分析。     

选择性双侧髂内动脉栓塞治疗异基因造血干细胞移植术后重度出血性膀胱炎

目的 探讨栓塞治疗在异基因造血干细胞移植（allo-HSCT）术后重度出血性膀胱炎（HC）中的应用价值。方法 6例接受allo-HSCT的患者在移植后24～80 d发生重度（Ⅲ～Ⅳ度）HC，经保守治疗无效，在数字减影血管造影（DSA）下行选择性双侧髂内动脉脏支栓塞术治疗。结果 6例患者共行8次栓塞治疗，4例治愈，1例好转，1例无效，有效率达83％。4例治愈的患者肉眼血尿消失时间为栓塞术后7～10 d，镜下血尿消失时间为栓塞术后20～30 d。6例患者均未出现严重并发症。结论 应用选择性双侧髂内动脉栓塞治疗重度HC是一种安全有效的措施，是治疗allo-HSCT后难治性、迁延不愈的重度HC的一种新的治疗手段。     

丁酸及其衍生物在血液病中研究和应用现状

本文综述了丁酸对血液肿瘤诱导分化作用，临床应用，药代动力学特点及作用机制等方面的研究概况。     

急性早幼粒细胞白血病中常见白血病基因突变的检测及其临床意义

本研究旨在探讨多种常见白血病基因突变是否协同参与急性早幼粒细胞白血病(APL)的发病机制,探讨这些突变与APL患者的临床表现、细胞遗传学及分子危险度分层的关系。收集了2005年2月至2010年1月收治的84例初治APL患者标本,采用基因组DNA-PCR方法分析骨髓单个核细胞中各个基因的突变情况及突变阳性患者的临床特征。结果表明,84例APL患者中合并突变的患者51例(60.7%),其中FLT3-ITD突变发生率最高,达到27.4%(23/84);其次,WT1突变12例,FLT3-TKD突变9例,TET2突变7例,N-RAS突变5例,ASXL1突变4例,EZH2突变2例,MLL-PTD、IDH1及CBL各突变1例;而JAK1、DNMT3、c-Kit、NPM1、IDH2、RUNX1、JAK2(V617F)等常见白血病相关基因未发现突变。结合临床资料,FLT3-ITD突变阳性患者的白细胞计数较高,N-RAS突变阳性患者的血小板计数较低,与野生型患者相比差异均有统计学意义(P<0.05);具有FLT3-ITD突变或N-RAS突变的患者总体生存期较无相关突变组患者明显缩短,差异有统计学意义(P<0.05)。单纯t(15;17)易位(69/84)患者与附加其他染色体异常(13/84)患者的总体生存时间无明显差别。结论:FLT3-ITD突变是APL患者常见的分子突变类型,其与N-RAS突变均为APL患者的不良预后因素;APL中合并其他染色体异常不影响患者的总体生存期。     

异基因造血干细胞移植治疗高度侵袭性非霍奇金淋巴瘤6例

高度侵袭性非霍奇金淋巴瘤(HGL)一般包括T、B或自然杀伤(NK)细胞来源的淋巴母细胞性淋巴瘤(LBL)和B细胞来源的Burkitt淋巴瘤(BL),也包括获得性免疫缺陷综合征(AIDS)相关的非霍奇金淋巴瘤(NHL)。此类淋巴瘤常发生于年轻人,疾病进展快,恶性程度高,预后差,容易复发,