CpG岛甲基化检测方法

目的：建立CpG岛甲基化测定方法,方法：收集20例急性粒细胞性白血病（AML）,18例多发性骨髓瘤（MM）,14例骨髓异常增生综合征（MDS）,15例慢性粒细胞性白血病（CML）及20 例正常对照组的外周血,分离单个核细胞,提取DNA,应用CpG岛思虑在化特异的方法（MSP－PCR）,测定P15和P16基因甲基化情况,结果：P15和P16基因甲基化在各种白血病的表率率分别为AML 80％和70％,MM72．2％和66．7％,MDS 57．1％和50％,CML0％,结论：P15及P16基因甲基化在AML,MM及MDS中有较高表达,而在CML中不表达。     

恶性血液病红细胞体积分布宽度的变化及意义

目的 :探讨恶性血液病患者红细胞体积分布宽度 (RDW )变化的临床意义。方法 :正常对照组 81例 ;恶性血液病例组 75例 ,其中急性白血病 5 7例、慢性粒细胞白血病 10例、多发性骨髓瘤 8例。采用血细胞自动分析仪检测研究对象外周血红细胞体积分布宽度和其它各种红细胞参数。结果 :红细胞体积分布宽度 (RDW)正常对照组为 13 36± 0 81,急性白血病组为 17 5 4± 2 63,慢性粒细胞白血病组为 2 0 99± 2 63,多发性骨髓瘤组为 2 1 31± 6 0 2 7。 3组恶性血液病患者与正常对照组相比RDW值明显升高 ,具有显著性差异 (P <0 0 1)。结论 :恶性血液病患者红细胞有质和量的异常变化。     

白血病间接免疫荧光法免疫表型分析

目的:研究白血病免疫表型的不同,明确诊断。方法:回顾性分析本院56例白血病患者免疫表型资料。结果:30例急性髓系白血病(AML)均表达髓系抗原,部分伴有淋巴抗原,但其阳性率明显低于急性淋巴细胞白血病(ALL)。7例M3的CD9抗原阳性率为100%,明显高于AML的其它亚型白血病。形态学诊断为ALL的12例白血病中,2例免疫表型为T-ALL,10例B-ALL。4例为杂合性急性白血病,它的诊断标准主要靠免疫分型。在10例慢性粒细胞性白血病(CML)中,4例发生急粒变,且急变期CD34抗原阳性率高于慢性期。结论:免疫表型分析与形态学、细胞化学三者结合为白血病的诊断提供了更可靠、更有价值的依据。     

三分类血液分析仪与血涂片镜检对白血病患者血液细胞分类结果差异分析

目的 探讨诊断白血病更准确、更全面的方法.方法 采用血液分析仪和手工涂片分类法同时诊断白血病.结果 14例ALL患者仪器不能进行白细胞分类和幼稚细胞提示;6例APL患者仪器进行了错误分类;24例AML患者仪器将血液中的幼稚细胞错误分类为小型白细胞;8例CML患者仪器将血液中的幼粒细胞及嗜酸嗜碱细胞分类为中间细胞群,以上均与手工分类有很大差异.结论 通过对56例白血病患者的血液分析,证实了在初诊白血病患者血样分析中,仪器不能给予白细胞分类及幼稚细胞的识别,对多形态、多变化性的血细胞识别有限.因此,用血液分析仪分析血样的同时,不能忽略血细胞形态学检查,防止幼稚细胞漏检.     

孤立性宫颈髓系肉瘤治疗后前庭大腺髓系肉瘤1例报道并文献复习

髓系肉瘤（Myeloid Sarcoma, MS）是一种发生于骨髓外的原始粒细胞或未成熟粒细胞形成的实体性肿瘤，患者发病主要与急性髓细胞白血病（AML）密切相关（白血病性），极少情况MS可发生在无AML病史（孤立性MS）。MS发病部位可累及全身任何部位，常见部位包括皮肤、淋巴结、睾丸、中枢神经系统及软组织和骨骼，但累及女性生殖道罕见。现报道1例发生于宫颈及前庭大腺的孤立性髓系肉瘤的诊治过程，并复习相关文献，以提高临床医生对该病的认识及警惕，同时为该病的诊断及治疗提供借鉴。     

半月科技要闻

国际新闻国际新闻基因突变引发致命白血病基因突变引发致命白血病一项新的研究表明,Ikaros基因突变在急性淋巴细胞白血病(ALL)的触发中扮演了重要角色。同样的费城染色体缺陷,慢性粒细胞白血病(CML)的患者能够通过治疗得到康复。而ALL患者几个月内会死亡,很难治愈。     

CML细胞抗原相关TCR Vα13／Vβ21和Vα18／Vβ21寡克隆T细胞及其CDR3序列特点

目的:分析慢性粒细胞白血病(CML)病人的克隆性增殖T细胞及其CDR3序列的特点。方法:利用RT-PCR和基因扫描分析1例CML患者外周血单个核细胞中的TCR Vα和Vβ基因的互补决定区(CDR3),了解各Vα和Vβ亚家族的限制性表达情况和T细胞克隆性增殖特点,寡克隆的PCR产物再进行序列分析。结果:该病人外周血T细胞表达18个TCR Vα和12个TCR Vβ亚家族,其中Vα13、Vα18、Vβ1和Vβ21亚家族呈寡克隆性。CDR3序列分析获得3个TCR克隆基因序列,分别为Vα13NJα49、Vα18NJα50和Vβ21NDβNJβ2.7。结论:获得1例CML病人外周血克隆性增殖αβ T细胞的3个TCR基因CDR3序列,提示病人可能存在与CML细胞抗原相关的Vα13/Vβ21或Vα18/Vβ21T细胞克隆。     

三聚氰胺的密度泛函理论研究

应用密度泛函理论(DFT)的B3LYP方法,在6-31G(d)基组水平上对三聚氰胺进行了研究。计算得到了分子的稳定构型,并对其进行了频率分析。然后利用Gaussview图形软件将频率分析数据转换为红外光谱。对红外光谱分析后发现,在400 cm-1～0 cm-1区域内分子的振动类型主要以分子内基团的整体摆动和胺基中C—H键的面外弯曲振动为主。在红外光谱的指纹区(1 333 cm-1～400 cm-1)谱线强度较弱,分子振动模式主要以弯曲振动为主,且存在七种分子的振动模式不具有红外活性。在光谱的特征谱带区(4 000 cm-1～1 333 cm-1),分子振动模式共有两种,即不对称伸缩振动和剪式振动。此外,整个红外光谱中振动峰的实际数目远小于简正振动的数目。     

丁基膦酸单丁酯稀土络合物的振动光谱研究

用稀土氯化物与丁基膦酸二丁酯反应制备了标题络合物Ln(BBP)3(Ln=La,Eu,Yb),测量了络合物4 000~100 cm-1的红外光谱和1 500~100 cm-1的拉曼光谱,对其主要红外吸收和拉曼谱带进行了归属.指认151cm-1的红外吸收带为Ln—O键的伸缩振动带,1 138 cm-1的红外吸收和1 136 cm-1的拉曼谱带为PO2基团的反对称伸缩振动带(aνsPO2),1 088 cm-1的红外吸收谱带为PO2基团的对称伸缩振动带(sνPO2).提出了络合物的每个稀土离子与邻近的三个稀土离子通过双—O—P(C4H9)(O C4H9)—O—桥相连接,形成“双桥二十四元环”多聚网络结构的假设模型.Ln—O键基本上为离子键.     

慢性粒细胞白血病患者FcεRⅠγ链基因表达上调

目的:在慢性粒细胞白血病病人(CML),CD3ζ链表达明显下降,分析与CD3ζ存在互补关系的FcεRⅠγ基因在CML患者中表达水平,以了解T细胞免疫中TCR信号转导的变化情况.方法:利用SYBR Green Ⅰ实时荧光定量RT-PCR法测定CML患者和健康人各20例的外周血单个核细胞(PBMCs)的FcεRⅠγ基因表达水平,以β2微球蛋白基因(β2MG)作为内参照,采用相对定量公式:2-△Ct×100％,计算FcεRⅠγ链的相对mRNA表达量.结果:CML患者PBMCs中FcεRⅠγ基因表达水平明显高于健康对照组(P＜0.001).FcεRⅠ γ表达水平变化与病人外周血CD3+T细胞比例无相关性.结论:CML病人中FcεRⅠ γ表达上升,提示FcεRⅠγ可能在一定程度调节CD3ζ链缺陷所带来的T细胞免疫异常.     

PCR检测白血病患者外周血中EB病毒的DNA

目的：了解白血病患者ＥＢ病毒感染情况。方法：收集２１例急性淋巴细胞白血病、１例慢性淋巴细胞白血病、１５例急性粒细胞白血病、８例慢性粒细胞白血病患者及３２例正常对照组的外周血,分离单个核细胞,提取ＤＮＡ,应用ＰＣＲ方法检测ＥＢ病毒ＤＮＡ。结果：在１例初诊慢性粒细胞白血病病人样本中发现ＥＢ病毒阳性,余均为阴性。结论：白血病患者存在ＥＢ病毒感染情况,但并不普遍。     

Viewpoints on the proinflammation state of leukemia

Proinflammation represents a pathophysiological state on the early stage of a number of diseases, especially the infectious and immunological ones. In recent years, proinflammation has attracted much attention, and the term 損roinflammation factors?appears frequently in the literature. While investigating leukemia and leukemic cells from the angle of 損roinflammation state? we got some intriguing findings, e.g. we detected the significantly elevated expression of proinflammation factor IL-18 in patients with acute myeloid leukemia (AML), which could up-regulate matrix metalloproteinases (MMP) and specific tissue inhibitors (TIMPs). The increased MMP may play a role in the aggressiveness of myeloid leukemic cells, and be associated with a poor prognosis. This phenomenon reflects an ignored aspect of leukemia. Investigations from the angle of 損roinflammation state?have broaden the fields of tumor and leukemia study.     

Impaired hydroxylation of 5-methylcytosine in myeloid cancers with mutant TET2

TET2 is a close relative of TET1, an enzyme that converts 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) in DNA. The gene encoding TET2 resides at chromosome 4q24, in a region showing recurrent microdeletions and copy-neutral loss of heterozygosity (CN-LOH) in patients with diverse myeloid malignancies. Somatic TET2 mutations are frequently observed in myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPN), MDS/MPN overlap syndromes including chronic myelomonocytic leukaemia (CMML), acute myeloid leukaemias (AML) and secondary AML (sAML). We show here that TET2 mutations associated with myeloid malignancies compromise catalytic activity. Bone marrow samples from patients with TET2 mutations displayed uniformly low levels of 5hmC in genomic DNA compared to bone marrow samples from healthy controls. Moreover, small hairpin RNA (shRNA)-mediated depletion of Tet2 in mouse haematopoietic precursors skewed their differentiation towards monocyte/macrophage lineages in culture. There was no significant difference in DNA methylation between bone marrow samples from patients with high 5hmC versus healthy controls, but samples from patients with low 5hmC showed hypomethylation relative to controls at the majority of differentially methylated CpG sites. Our results demonstrate that Tet2 is important for normal myelopoiesis, and suggest that disruption of TET2 enzymatic activity favours myeloid tumorigenesis. Measurement of 5hmC levels in myeloid malignancies may prove valuable as a diagnostic and prognostic tool, to tailor therapies and assess responses to anticancer drugs.     

50例急性白血病细胞电镜研究

用电镜和光镜相结合观察50例急性白血病,两者诊断符合率达90%,电镜有助于急性白血病的诊断分型,SEM 下急粒以嵴样型细胞为多,占48～90%;急单中的皱膜型细胞为70～81%;急淋白血病细胞表面特征主要有光滑型和微绒毛型两种.TEM 观察结果和既往作者报道的相似.     

核仁抗原在白血病诊断中的初步观察

用抗核仁抗原NAg-1的抗血清对各种白血病人骨髓组织进行免疫荧光染色.结果表明,急性白血病患者的NAg-1阳性细胞率显著高于正常人及各种非白血病贫血患者.在处于完全缓解的白血病患者中,具有较高的NAg-1阳性率者(>10％)比较低者(<10％)更易复发.     

IDH1(R132H) mutation increases murine haematopoietic progenitors and alters epigenetics

Mutations in the IDH1 and IDH2 genes encoding isocitrate dehydrogenases are frequently found in human glioblastomas and cytogenetically normal acute myeloid leukaemias (AML). These alterations are gain-of-function mutations in that they drive the synthesis of the ‘oncometabolite’ R-2-hydroxyglutarate (2HG). It remains unclear how IDH1 and IDH2 mutations modify myeloid cell development and promote leukaemogenesis. Here we report the characterization of conditional knock-in (KI) mice in which the most common IDH1 mutation, IDH1(R132H), is inserted into the endogenous murine Idh1 locus and is expressed in all haematopoietic cells (Vav-KI mice) or specifically in cells of the myeloid lineage (LysM-KI mice). These mutants show increased numbers of early haematopoietic progenitors and develop splenomegaly and anaemia with extramedullary haematopoiesis, suggesting a dysfunctional bone marrow niche. Furthermore, LysM-KI cells have hypermethylated histones and changes to DNA methylation similar to those observed in human IDH1- or IDH2-mutant AML. To our knowledge, our study is the first to describe the generation and characterization of conditional IDH1(R132H)-KI mice, and also the first report to demonstrate the induction of a leukaemic DNA methylation signature in a mouse model. Our report thus sheds light on the mechanistic links between IDH1 mutation and human AML.     

Altered adhesive interactions with marrow stroma of haematopoietic progenitor cells in chronic myeloid leukaemia

Normal haematopoietic cell regulation involves interaction between marrow stromal cells and haematopoietic progenitor cells which may be facilitated by specific recognition and adhesion. Some leukaemogenic events might produce a selective growth advantage by altering this regulatory network, possibly by diminishing the capacities of cells to adhere to stromal elements. Using an in vitro culture system which allows investigation of adhesion to stromal layers and subsequent colony formation by blast colony-forming cells (B1-CFC) in normal marrow and Ph+ chronic myeloid leukaemic (CML) blood, we compared the adhesive properties of normal and malignant progenitor cells. We present evidence that altered adhesive interactions between primitive progenitor cells and marrow stromal cells occur in CML.     

WT1联合多参数流式在急性髓系白血病预后的临床观察

目的 研究WT1基因定量联合多参数流式（FCM）在急性髓系白血病（AML）预后的临床观察。方法62例AML患者分为低危组、中危组、高危组;治疗上参照《成人急性髓系白血病（非急性早幼粒细胞白血病）中国诊疗指南2017年版》,用RT-qPCR方法测定患者WT1基因表达水平;同时用多参数流式细胞分析技术（FCM）分析患者低水平微小残留（MRD）水平;比较不同预后患者WT1表达及WT1表达与预后的关系。观察WT1基因定量联合FCM对AML患者预后评估的临床意义。对患者进行随访不低于2年。结果 AML患者WT1高表达比例为77.42%（48/62）,不同预后分型的AML患者WT1高表达有统计学差异（P <0.05）。在中、高危组的患者WT1高表达明显高于低危组。初发的WT1高表达与WT1低表达患者在诱导缓解率无统计学意义（P >0.05）,但WT1高表达患者的2年无病生存率、2年总生存率均WT1低表达患者,差异有统计学意义（35.41% vs 71.43%,47.92% vs 85.71% P < 0.05）。WT1基因联合FCM预测AML早期复发的敏感性、特异性均高于单独WT1基因和单独的FCM（P < 0.05）。结论 WT1在AML患者中高表达,与患者预后相关,联合FCM可有效的预测患者早期复发,可作为临床治疗及预后判断的靶点。     

FTIR-HATR诊断α-地中海贫血

将傅里叶红外光谱与水平衰减全反射技术(FTIR-HATR)结合,对α-地中海贫血样品与正常样品进行检验,通过改进的相对强度计算方法进行对比分析.结果表明:α-地贫组与正常组间在I107t/I1152、I1517/I1074、I3295/I3372相对强度上存在显著差异;在峰的指认上,波数1074cm-1、1152cm-...