幼淋白血病超微结构观察附一例报告

幼淋细胞白血病(Prolymphocytic Leukaemia)(PLL)国内外均有数篇报导,是以脾巨大,常用治疗慢淋的药物疗效差,幼淋细胞占优势的淋巴细胞恶性增殖性疾患。对其超微结构的研究国外已有少数报导。我们对一例慢淋幼淋变病人骨髓中的幼淋细胞用TEM观察其超微结构并与光学显微镜下瑞氏染色涂片的形态进行了对比。病历摘要:刘×、女性、40岁,四年前因牙龈出血、乏力住某院,经骨穿活检诊断为:慢性淋巴细胞白血病(CLL)。按CLL治疗最初有效,但近来脾脏明显肿大,化疗无效,疑幼淋细胞白血病或慢淋幼淋变于1984年2月转入我院。体检:贫血貌,全身浅表淋巴结为黄豆至枣大,质硬无触痛,脾肋下23cm,质硬边钝,表面光滑无     

细支气管肺泡癌患者EGFR基因突变的研究

目的探讨表皮生长因子受体（EGFR）基因酪氨酸激酶域体细胞在细支气管肺泡癌（BAC）患者中突变的相关因素。方法分析和检测本院37例细支气管肺泡癌石蜡包埋标本EGFR基因突变状况,采用PCR技术进行EGFR基因18、19、20和21外显子突变分析。结果37例BAC患者中有18例（48．6％）酪氨酸激酶域存在体细胞突变,其中3例（8．1％）为18外显子替代突变,5例（13．5％）为19外显子突变,7例（18．9％）为20外显子替代突变,8例（21．6％）为21外显子替代突变。结论EGFR基因与BAC密切相关。     

白血病性淋巴细胞免疫标记分型及超微结构研究

用羊红细胞和酵母菌单独标记法及混合花环法研究了18例(急淋13例,慢淋5例)淋巴细胞型白血病的分型,同时按FAB分型标准进行了分型,并用透射电镜观察了各型白血病细胞的超微结构特征。 18例患者,B淋巴细胞型7例(5例慢淋,2例急淋)T淋巴细胞型2例,9例N细胞各型中还见少数D细胞(双标记)。     

慢性粒细胞性白血病血小板的电镜观察

慢性粒细胞性白血病是一种单克隆骨髓增生性疾病,起源于多能干细胞的肿瘤样转变。尽管发病时血小板数量增多,但仍常伴有出血。本组观察7例慢粒患者血小板,发现其超微结构明显异常:①细胞大小悬殊,多数为2μ以下的小细胞,但同时常见5至10μ的巨大血小板;②多数细胞胞浆致密性改变,而正常应具中等电子密度的     

白血病的电镜诊断

应用电镜分别对5例不同类型白血病患者骨髓穿刺细胞进行了观察,观察结果表明例一、二均为急性早幼粒细胞白血病,两例骨髓细胞中均见有许多异常早幼粒细胞胞浆内含有Auer小体,但两例细胞内Auer小体结构呈明显差异,例三、四为不同类型毛细胸白血病,即典型毛细胞白血病（Ⅰ型）及变异型毛细胞白血病（Ⅱ型）,两型细胞在结构上的差异主要表现为细胞膜表面毛样突起的长度及数量不同,变异型毛细胞白血病细胞的特点仅在电镜下才能辨认。例五为急性粒细胞白血病的部分分化型,其中早幼粒细胞胞浆内仅具极少数小的电子致密颗粒,电镜对该种细胞的分辨显示出光镜所不具有的优势,并提示电镜在白现诊断中的重要意义。     

趋化因子配体18与鼻咽癌相关性研究及意义

目的：探讨趋化因子配体18（CCL18）在鼻咽癌患者血浆中的表达水平及检测在体外对人鼻咽癌细胞株增殖和迁移能力的影响。方法：收集到重庆医科大学附属第二医院就诊并病理诊断为鼻咽癌患者37例,在未接受任何治疗前留取血液标本,另取20例健康体检者外周血液标本作为正常人群对照。以酶联免疫吸附测定（ELIAS）方法检测血浆中CCL18的表达水平。体外实验以人鼻咽癌细胞株HNE-1为研究对象,用噻唑蓝（MTT）比色法检测重组CCL18对细胞增殖能力的影响,用Transwe11体外迁移系统检测CCL18对鼻咽癌细胞的趋化作用。结果：鼻咽癌患者血浆中CCL18的表达水平明显高于健康体检者（P〈0．0001）,伴有颈部淋巴结转移的鼻咽癌患者较未发生颈部淋巴结转移者血浆中CCL18的表达水平显著升高（P〈0．05）。CCL18因子在体外对鼻咽癌细胞无明显促进增殖作用（P〉0．05）。CCL18对鼻咽癌细胞具有明显促进迁移趋化作用,并呈现浓度依赖性。结论：CCL18是鼻咽癌的生物学标志物,其表达与鼻咽癌的侵袭转移相关,对鼻咽癌的诊断和预后判断有一定临床意义。     

氟达拉滨联合利妥昔单抗治疗高龄慢性淋巴细胞白血病

目的考察氟达拉滨联合治疗高龄慢性淋巴细胞白血病患者的有效性和安全性。方法应用氟达拉滨联合利妥昔单抗方案治疗11例患者,根据患者情况分次给药,25~30mg/m2隔日或每周1~2次静脉注射,或口服剂型10mg~20mg/d连续用药,观察患者应用氟达拉滨过程中及其后的不良反应及疗效。结果全组11例患者CR+PR为10例,其中CR6例,PR4例,NR1例。不良反应主要为粒细胞及血小板减少等骨髓抑制,未出现严重感染,无化疗相关死亡。结论氟达拉滨联合利妥昔单抗治疗高龄慢性淋巴细胞白血病具有较高的安全性和有效性。     

XE-5000检测网织血小板在血液疾病诊断中的应用

目的探讨XE-5000全自动血液分析仪检测网织血小板在血液疾病中的应用。方法随机选择特发性血小板减少性紫癜(ITP)35例,再生障碍性贫血(AA)28例,急性白血病(AL)40例,慢性粒细胞白血病(CGL)24例,应用XE-5000全自动血液分析仪测定网织血小板百分比RP%,网织血小板绝对值IPF(×109/L)和平均血小板体积MPV。结果ITP初诊组,CGL组IPF%、MPV明显高于正常对照组(P<0.05),AA组,AL组患者IPF%高于正常对照组但无显著性差异(P>0.05)。ITP缓解组IPF%低于初诊组(P<0.05),与正常对照组比较无显著性差异(P>0.05)。结论外周血网织血小板数量可反映骨髓巨核系统生成血小板的情况,从而用于疾病的诊断和鉴别诊断。XE-5000全自动血液分析仪是一种实用和有效的IPF检测仪器。     

丹参酮ⅡA增效大鼠骨髓源性内皮祖细胞VEGF、SDF-1表达

目的：研究在丹参酮ⅡA的辅助下,骨髓源性内皮祖细胞（EPCs）VEGF与SDF-1表达是否能增强,从而促进细胞归巢。方法：Percoll密度梯度离心法分离SD大鼠骨髓单个核细胞,血管内皮生长因子（VEGF）、表皮生长因子（（EGF）、碱性成纤维细胞生长因子（bFGF）诱导培养,并进行形态学、免疫学（免疫细胞化学染色）、功能学（Dil-acLDL与FITC-UEA-1双荧光染色）鉴定。将鉴定为EPCs的细胞分为单纯EPCs组（对照组）、EPCs＋丹参酮ⅡA组（加药组）。细胞培养第9d,Western blot与实时荧光定量PCR检测各组VEGF、SDF-1基因与蛋白表达。结果：EPCs＋丹参酮IIA组VEGF、SDF-1基因与蛋白表达均高于EPCs组（p均〈0.05）。结论：丹参酮ⅡA可上调大鼠骨髓源性内皮祖细胞VEGF及SDF-1表达,从而可能在缺血环境中更好促进EPCs归巢,更有效促进血管修复与再生。     

5-ALA介导的光动力治疗在耐药白血病中的作用机制研究

目的: 本研究观察了5-氨基乙酰丙酸介导的光动力治疗(ALA-PDT)对耐药白血病细胞株HL-60/ADR的杀伤作用及对耐药白血病原代细胞存活率的影响。方法: 以耐药白血病细胞株HL-60/ADR为实验模型, 同时在11例白血病患者原代细胞中进行检测。实验分为4组, 对照组、单纯ALA组、单纯光照组及ALA＋PDT组。用MTT法测定细胞的存活率, 采用阳离子脂质荧光探针JC-1检测线粒体跨膜电位, 用Real-time PCR检测PDT前后HL-60/ADR细胞株中bcl-2基因及多药耐药基因MRP的表达变化。结果: ALA-PDT后HL-60/ADR细胞株线粒体跨膜电位出现快速下降, 0, 2, 4 h时线粒体跨膜电位崩塌的细胞比例分别升高至55.91%±2.60%、64.27%±1.08%、82.17%±0.43%, 与对照组相比皆有显著差异(P＜0.05), 呈时间依赖性。而单纯ALA组和单纯光照组则无明显变化。HL-60/ADR细胞株在ALA-PDT后Bcl-2和MRP基因均呈明显下降趋势。在初发和复发难治急性白血病原代细胞中ALA＋PDT组均显示较强的光动力效应。结论: ALA-PDT诱导的HL-60/ADR细胞的杀伤可能与其影响线粒体跨膜电位有关, 即通过影响线粒体功能促进细胞凋亡。同时表明ALA介导的光动力作用部分是通过在基因转录水平下调抗凋亡基因Bcl-2而促进凋亡的发生, 另一方面通过下调耐药基因MRP的表达而部分逆转耐药。同时ALA-PDT对原代白血病细胞同样有较大的抑制作用。     

Clonogenic cultivation of bone marrow fibroblast precursors in human myeloproliferative diseases

Techniques of clonogenic cultivation with the application of xenogenous feeder (rabbit irradiated bone marrow) were used to study a number of bone marrow colony-forming cells (CFU-F) in 70 patients. A significant increase of CFU-F is observed in chronic myelocytic leukemia and in hepatosplenomegalies of non-leukemic origin CFU-F decreases considerably in the cases of myelofibrosis. Trypsinisation of the bone marrow taken from the cases of myelofibrosis results in a sharp CFU-F increase.     

Monoclonal antibodies ICO-02 to blast cell antigens in patients with chronic myeloleukemia in blast crisis

Mice were immunized with blood cells of a patient with chronic granulocytic leukemia, and their cells were subsequently used for the preparation of hybridoma ICO-02. This hybridoma is continuously producing monoclonal antibodies which reacted with cells in 4 out of 13 patients with blastic crisis of chronic granulocytic leukemia and in 6 out of 38 patients with acute lymphoblastic leukemia. Antibodies reacted with blast cells in 2 out of 3 patients with undifferentiated blastic crisis of chronic myelocytic leukemia and in 2 out of 5 patients with lymphoid variant of blastic crisis of chronic granulocytic leukemia. Cells of 6 patients with acute lymphoblastic leukemia which reacted with the monoclonal antibodies had immunological markers of T lymphocytes bone-marrow precursors. Monoclonal antibodies did not react with cells of blood and bone marrow from healthy people and from patients with chronic lymphocytic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute monoblastic leukemia and lymphosarcoma.     

The effect of the bone marrow fibroblasts from patients with malignant diseases of the blood system on the colony-forming capacity of granulomonocytic precursor cells

The inhibiting activity of bone marrow fibroblasts and their ability to sustain survival of granulocytic cell precursors have been studied in patients with chronic myelocytic leukemia, chronic lymphoproliferative disorders, acute leukemia, myelodysplastic syndrome. Fibroblast conditioned medium inhibits proliferation of granulomonocytic precursors stimulated by leucocyte feeder layer or leucocyte conditioned medium. The effect depends both on the presence of monocytes and the specificity of the disease. The inhibitory effect is related with long-ranged factors. Bone marrow fibroblasts are able to sustain survival of hemopoietic precursors via humoral and cell-cell mechanisms. The regulatory role of bone marrow fibroblasts in hemopoietic disorders is discussed.     

ICO-10 monoclonal antibodies to the Thy-1 antigen

Mouse monoclonal antibodies (MAB) ICO-10 to Thy-1 antigen were obtained. MAB ICO-10 reacted in indirect immunofluorescence test with 5.7 +/- 0.8% human thymocytes. Antibodies did not react with granulocytes, monocytes, T- and non-T cells from peripheral blood, and with marrow cells of healthy donors. MAB ICO-10 reacted with blast cells from 25 of 53 patients with T-cell acute lymphoblastic leukemia (ALL), from 2 of 5 patients with B-cell ALL. This antigen was absent on blood and marrow cells from some patients with ALL, 80 patients with chronic lymphoid leukemia, 54 patients with chronic granulocytic leukemia at the stage of blastic crisis, 128 patients with acute nonlymphoblastic leukemia. Antibodies are specifically bound to thymocytes and spleen cells of Thy 1.1 and Thy 1.2 mice. MAB ICO-10 detect Thy-1 antigen expressed on human hematopoietic cells. MAB ICO-10 may be applied for human leukemia and lymphoma immune diagnosis.     

Electric fields in bone marrow substructures at power-line frequencies

Bone marrow is known to be responsible for leukemia. In order to study the hypothesis relating power-line frequencies electromagnetic fields and childhood leukemia from a subcellular perspective, two models of bone marrow substructures exposed to electric field are computed numerically. A set of cancellous bone data obtained from computed tomography scan is computed using both the finite element method (FEM) and scalar potential finite difference method. A maximum electric field enhancement of 50% is observed. Another model of bone marrow stroma cells is implemented only in FEM using thin film approximation. The transmembrane potential (TMP) change across the gap junctions is found to range from several to over 200 microV. The two results suggest that imperceptible contact currents can produce biologically significant TMP change at least in a limited number of bone marrow stroma cells.     

PDT对反义bcr-abl寡核苷酸k562细胞转染的影响

目的 :体外研究光动力治疗 (PDT)对b3a2型反义bcr-abl寡核苷酸 (ASO)慢性髓细胞性白血病 (CML)细胞株K5 6 2转染的影响 ,为PDT合并反义技术应用于CML患者治疗提供实验基础。方法 :半导体激光 ,波长 6 5 0nm ,剂量 9J cm2 垂直照射。光敏剂为血卟啉单甲醚(HMME) ,采用体外细胞培养技术 ,流式细胞仪检测PDT实验组与对照组荧光标记的反义bcr-abl寡核苷酸 (ASPO)K5 6 2细胞摄入率及荧光强度。结果 :①PDT可显著提高k5 6 2细胞对反义bcr-abl寡核苷酸的摄入 ,比直接转染增加转染率可达 4-1 0倍。且k5 6 2细胞对反义bcr-abl寡核苷酸的摄入和反义bcr-abl寡核苷酸浓度及作用时间有关。②光敏剂量浓度为 0 .9μmol·L- 1 时转染的效率最高 ,4小时点细胞内平均荧光强度最强 (P <0 .0 1 ) ,且细胞内平均荧光强度随ASPO浓度的增加而增高。但直接转染时 ,6小时才达到高峰。浓度为 0 .6 μmol·L- 1 ～ 0 .9μmol·L- 1 时K5 6 2细胞对荧光物质的摄入即达饱和。结论 :PDT可以增加反义bcr -abl寡核苷酸K5 6 2细胞转染率     

露蜂房蛋白对急性髓细胞白血病患者骨髓单个核细胞超微结构的影响

目的：观察中药露蜂房蛋白(NVP)成份对急性髓细胞白血病(AML)患者骨髓单个核细胞(BMMNC)超微结构的影响。方法：采用体外细胞培养,加入不同浓度露蜂房蛋白成分作用于AML患者BMMNC,培养72h收集细胞,常规制备超薄切片,透射电子显微镜下观察AML患者BMMNC的超微结构。结果：露蜂房蛋白各处理组细胞核染色质浓缩、边集,呈新月形或环状或细胞核碎裂呈块状。线粒体出现空泡样变及髓样变。结论：中药露蜂房蛋白成份有明显诱导AML患者体外培养BMMNC凋亡的作用,并呈现典型的细胞凋亡超微结构改变。     

Block of NK cell activity by ICO-11 monoclonal antibodies

Antigen expression determined by ICO-11 monoclonal antibodies was studied on leukemia cells from bone marrow, blood cells from patients with lymphoproliferative diseases and natural killer (NK) cells responsible for natural resistance. ICO-11 monoclonal antibodies were shown to recognize the antigen expressed on NK-cells, predecessors of T-cells (thymocytes), myelomonocytes (myeloblasts, monoblasts) and to block NK-cell activity.     

Colony-forming ability of bone marrow hematopoietic cells in mice with transplanted leukemia during administration of carminomycin

Effects of carminomycin on the colony-forming ability of bone marrow haemopoietic stem cells (CFUs) were compared in normal mice with steady state and actively regenerating bone marrow, and in P-388 and La leukemia-bearing mice. CFUs assays were performed 24 h after treatment of donor mice with the increasing doses of the drug. Leukemia-bearing mice received carminomycin on the 5th day after transplantation. The dose-effect curves were exponential for CFUS normal mice with both the steady state and active proliferation state of bone marrow. The maximal effect was found 24 h after injection of 0.7 mg/kg of carminomycin (ED50 for CFUS) being more pronounced in regenerating bone marrow. The dose-effect curves for leukemias were also exponential. In the case of La leukemia the killing of CFUs by carminomycin was the highest as compared with P-388 leukemia.     

侵袭性NK细胞白血病1例报告并文献复习

目的提高对侵袭性NK细胞白血病（ANKL）的认识。方法报道1例侵袭性NK细胞白血病的诊断及其治疗经过,并进行文献复习。结果侵袭性NK细胞白血病多见于亚洲人;发病年龄相对较轻;临床常表现为高热、盗汗、肝脾淋巴结骨髓受侵犯、肝功能不全、反应性噬血细胞综合征、血细胞减少等多系统受累,免疫表型表达CD2、CD7、CD16、CD56,预后差,生存时间多以周来计算。结论ANKL是一种少见的恶性白血病,临床有疑似病例时应尽快完善骨髓、免疫表型、病理活检、基因重排等相关检查,及早明确诊断,争取治疗时间。