急性髓细胞性白血病患者外周血单个核细胞CD28 mRNA的表达

背景：大量研究显示,肿瘤患者外周血T细胞表面共刺激分子CD28蛋白表达存在差异,提示共刺激通路异常可能与恶性肿瘤的发生进展有关。 目的：观察急性髓细胞性白血病外周血单个核细胞共刺激信号分子CD28 mRNA在中的表达。 方法：急性髓细胞性白血病患者80例,其中M0型7例,M1型6例,M2型18例,M3型15例,M4型17例,M5型9例,M6型8例。并根据急性白血病疗效标准将80例患者分为完全治愈组、缓解组、未缓解组。采用Taqman探针实时荧光定量PCR检测80例患者及76名健康人群外周血单个核细胞CD28 mRNA的表达。 结果与结论：急性髓细胞性白血病外周血单个核细胞M1,M3和M4亚型中的CD28 mRNA表达量低于健康人群 (P < 0.05);急性髓细胞性白血病未缓解组中CD28 mRNA低于健康人群 (P < 0.05),完全治愈组和缓解组中CD28 mRNA表达与健康人群差异无显著性意义。说明急性髓细胞白血病患者外周血单个核细胞存在CD28 mRNA表达缺陷,并与临床分期、病情进展及预后有关。     

CD56抗原表达在急性髓细胞性白血病并发中枢神经系统浸润中的意义

目的 探讨CD56抗原表达在急性髓细胞性白血病(AML)并发中枢神经系统浸润(CNSL)中的意义及髓外浸润的机制。方法 回顾性分析2012-01—2018-12郑州市第一人民医院收治的86例连续接受全身化疗的AML患者CD56表达的临床特征,将CD56⁺ AML患者的分类数据与CD56-AML患者的分类数据进行比较。结果 86例患者中26.7%的患者表达CD56抗原,其表达与CD13⁺、CD34⁺、CD38^-和CD11b⁺免疫表型显著相关。全身化疗后,11.6%的患者中枢神经系统受累,多集中发生在AML完全缓解(CR)后6～12个月,发生率达76.9%。CD56抗原阳性患者的3 a无病生存期(DFS)明显低于抗原阴性患者(21.7%对47.6%,P=0.04)。结论 CD56抗原表达是AML并发中枢神经系统浸润的重要危险因素。CD56抗原阳性患者DFS与OS短于阴性患者。     

中药复方对急性髓细胞性白血病干细胞Flt3和N-ras基因表达的影响

背景：在白血病患者体内存在一群白血病干细胞,这些恶性干细胞是白血病细胞存在和不断增殖的根源,针对干预白血病干细胞的特异性靶向治疗药物已成为近年来研究热点。 目的：探讨中药复方对急性髓细胞性白血病干细胞Flt3和N-ras基因表达的影响。 设计、时间及地点：随机区组实验,于2007-09/2008-12在山东中医药大学附属医院完成。 对象：2006-2007年山东中医药大学附属医院、山东大学齐鲁医院血液科就诊的急性髓细胞性白血病初治患者50例,FAB分型M1 5例,M2 15例,M4 9例,M5 21例。 方法：原方：黄芪、白花蛇舌草、小蓟、太子参、半枝莲、蒲公英、生地、黄精、女贞子、旱莲草、天冬、麦冬、白术、茯苓、甘草。扶正方：黄芪、太子参、生地、黄精、女贞子、天冬、麦冬、白术、茯苓、甘草。祛邪方：白花蛇舌草、小蓟、半枝莲、蒲公英、旱莲草。制剂：上述3组组方药物由山东中医药大学附属医院中药制剂实验室(国家三级重要制剂实验室)配制成1 g/mL的药液,无菌试验阴性后过滤除菌备用。常规无菌采集急性髓细胞性白血病M1,M2,M4,M5型患者骨髓各5 mL,稀释后加入淋巴细胞分离液,联合应用免疫磁珠分选系统和流式细胞仪分离纯化白血病干细胞。调整细胞浓度至2×108 L-1,均分成4组：对照组不加药物,实验组分别加入原方、扶正方、祛邪方的对应中药制剂,终浓度为100 mg/L,继续培养48 h后进行指标检测。 主要观察指标：RT-PCR法检测Flt3、N-ras基因的表达。 结果：与对照组比较,原方组、扶正方组、祛邪方组Flt3表达阳性率均明显降低(P < 0.05),但原方组降低幅度明显大于扶正方组、祛邪方组(P < 0.05);扶正方组与祛邪方组Flt3表达阳性率比较无明显差异(P > 0.05)。4组N-ras表达阳性率与Flt3表达情况基本一致。 结论：Flt3和N-ras基因在白血病干细胞中存在过度表达,中药原方及其拆方后的扶正方、祛邪方均能够降低白血病干细胞中Flt3和N-ras基因的表达水平。     

融合蛋白AML1-ETO对C/EBPα基因表达的影响☆

背景：已有研究显示AML1-ETO可以通过包括抑制C/EBPα的功能在内的机制引起分化的阻滞。 目的：观察AML1-ETO对髓系分化相关基因C/EBPα表达的影响，探讨其在白血病发生中的作用。 方法：应用流式细胞术检测急性单核细胞白血病细胞株U937、U937-MOCK和U937-A/E1细胞表面抗原CD11b、CD14 的表达；荧光实时定量PCR检测C/EBPα mRNA的表达；免疫印迹法检测C/EBPα蛋白的表达；染色质免疫沉淀技术研究转染细胞中AML1-ETO与C/EBPα基因启动子之间直接的相互作用情况。 结果与结论：AML1-ETO转染细胞的细胞表面分化抗原CD11b 和CD14的表达明显下降，且转染了AML1-ETO的U937细胞系中，C/EBPα的mRNA与蛋白水平均下调，转染细胞沉淀富集的DNA中含有C/EBPα基因的启动子序列。结果提示C/EBPα是AML1-ETO 的直接靶基因，AML1-ETO能下调C/EBPα的表达。     

免疫分型及临床应用

血液系统疾病与白细胞分化抗原相关，白细胞分化抗原是存在于白细胞不同谱系，不同分化阶段以及激活过程中出现或消失的细胞表面抗原分子，可作为不同细胞谱系的表面标志。流式细胞术已成为诊断血液系统恶性肿瘤不可缺少的工具，广泛应用于白血病的临床检测。免疫分型是免疫治疗的必要前提，在白血病的分型中具有越来越重要的作用。通过对细胞抗原分子研究的不断深入，分子生物学等相关技术的发展，免疫学分型在白血病、多发性骨髓瘤、再生障碍性贫血及阵发性睡眠性血红蛋白尿诊断、治疗及预后中必将起到越来越重要的作用。     

急性白血病骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D的表达☆

背景：据作者查新检索,国内外有关急性白血病骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D活性及mRNA表达与白血病类型、患者肝脾淋巴结肿大关系的报道罕见。 目的：探讨糖基化磷脂酰肌醇特异性磷脂酶D的表达与急性白血病类型、急性髓系白血病患者肝脾淋巴结肿大的关系。 方法：急性髓系白血病组骨髓标本43例,急性淋巴细胞白血病组骨髓标本28例,以21例健康志愿者作为正常对照组。密度梯度离心法获得骨髓单个核细胞,用GPI锚定的胎盘碱性磷酸酶作为底物,Triton-X114分相法检测糖基化磷脂酰肌醇特异性磷脂酶D的活性,半定量RT-PCR法检测糖基化磷脂酰肌醇特异性磷脂酶D mRNA的表达,并分析糖基化磷脂酰肌醇特异性磷脂酶D活性和mRNA表达水平与急性白血病类型、急性髓系白血病患者肝脾淋巴结肿大的关系。 结果与结论：与正常对照组比较,急性髓系白血病组骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D活性和mRNA表达均明显升高(P < 0.01);急性淋巴细胞白血病组骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D活性和mRNA表达均明显降低(P < 0.01)。在43例急性髓系白血病患者中,13例患者检查发现有肝脾淋巴结肿大,30例患者检查无肝脾淋巴结肿大,无肝脾淋巴结肿大标本骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D活性和mRNA表达均明显高于有肝脾淋巴结肿大标本(P < 0.05)。结果证实急性白血病骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D的活性与mRNA变化一致,其表达水平与急性白血病类型及急性髓系白血病患者有无肝脾和/或淋巴结肿大有关。     

应用流式细胞术测定CD24表达鉴定人髓核细胞

背景：CD24是成熟髓核细胞表面表达的特异性分子，测定CD24的表达是否可以作为鉴定髓核细胞的一种方法？ 目的：验证流式细胞仪测定髓核细胞CD24表达鉴定髓核细胞的方法。 方法：采用酶消化法分离培养人髓核细胞，取生长旺盛的第2代人髓核细胞爬片进行Ⅱ型胶原免疫组织化学染色，RT-PCR测定Ⅱ型胶原及SOX9的表达，流式细胞仪检测其CD24阳性表达率，分析来自同一批细胞的免疫组织化学染色、RT-PCR结果与CD24阳性率的相关性。 结果与结论：体外分离培养的7例第2代髓核细胞爬片Ⅱ型胶原免疫组织化学染色呈阳性，RT-PCR结果阳性，流式细胞仪测定CD24其阳性率均为50%以上，髓核细胞分泌胞外基质Ⅱ型胶原与SOX9的表达与CD24阳性率结果有相关性。提示采用流式细胞仪测定CD24表达可以快速、简便地鉴定髓核细胞。     

细胞黏附分子CD99在人髓母细胞瘤中的表达及意义

目的 探讨细胞黏附分子CD99在人髓母细胞瘤组织中的表达特点及其临床意义。方法 采用免疫组化法检测107例髓母细胞瘤及15例瘤旁正常脑组织标本中CD99的表达。所有病人均随访至死亡或截止至2013年12月，随访时间为12~80个月，利用Kaplan-Meier生存曲线进行生存分析。结果 髓母细胞瘤CD99阳性表达率（57.94%，62/107）明显高于瘤周正常脑组织（0%，0/15；P<0.05）。CD99的表达水平与病人性别、年龄、肿瘤的病理组织学分型和分子亚型均无明显相关性（P>0.05）。CD99的高表达与髓母细胞瘤病人的预后呈显著负相关（P<0.05）。结论 CD99高表达与髓母细胞瘤预后显著相关，可作为预后判断的指标及治疗髓母细胞瘤潜在的新靶点。     

建立人急性粒细胞白血病M2裸鼠模型***

背景：人实体瘤细胞容易在小鼠体内成瘤,但人荷瘤白血病模型很难构建。通过放射线或环磷酰胺抑制裸鼠免疫系统预处理,可构建低成本、高稳定性的裸鼠模型。 目的：探讨融合基因AML/ETO阳性的人急性粒细胞白血病M2白血病细胞Kasumi-1在BALB/c裸鼠体内建立白血病模型的方法。 方法：将BALB/c裸鼠以抽签法随机分3组：环磷酰胺组腹腔注射环磷酰胺连续2 d后,尾静脉注射Kasumi-1细胞8×105/只;照射组给予X射线全身照射,照射当天尾静脉注射Kasumi-1细胞;无预处理组未作任何处理,尾静脉注射Kasumi-1白血病细胞。另取3只正常BALB/c裸鼠作为正常对照。检测外周血涂片、骨髓涂片,流式细胞仪检测骨髓细胞免疫分型,RT- PCR 检测白血病细胞瘤负荷,FISH检测骨髓细胞AML/ETO融合基因阳性细胞百分比。 结果与结论：未经任何预处理裸鼠建模14 d血涂片中白血病细胞达3.5%,骨髓中肿瘤细胞百分比可达40%以上,与FISH和流式细胞仪检测白血病细胞比例一致,且随着接种时间延长,瘤负荷不断增加。全身照射和环磷酰胺注射后的裸鼠瘤负荷高于无预处理组,但仍可带瘤生存60 d。正常裸鼠外周血单个核细胞RT-PCR未发现有融和基因AML/ETO,其他3组均可见融和基因AML/ETO的mRNA表达。提示给予环磷酰胺组和X线照射预处理或单纯尾静脉接种Kasimi-1细胞均可建立急性粒细胞白血病M2裸鼠模型。     

急性髓系白血病患者外周血单个核细胞Toll样受体9以及血清肿瘤坏死因子和凋亡基因FAS的表达

背景：Toll样受体9(TLR9)、肿瘤坏死因子a、Fas可能共同参与白血病的发生发展过程。 目的：测定TLR9在急性髓系白血病患者外周血单个核细胞中的表达及血清肿瘤坏死因子α、Fas水平。 方法：从急性髓系白血病患者与正常对照组中分离出外周血单个核细胞，采用反转录-聚合酶链反应法检测外周血单个核细胞中TLR9 mRNA的表达水平，采用酶联免疫吸附试验法检测血清肿瘤坏死因子α、Fas水平。 结果与结论：在急性髓系白血病患者初治组和难治复发组中，外周血单个核细胞中TLR9 mRNA表达高于正常对照组(P < 0.01)，化疗后完全缓解组与正常对照组差异无显著性意义(P > 0.05)；各病例组血清肿瘤坏死因子α、Fas水平显著高于正常对照组(P < 0.01)。TLR9 mRNA的表达与血清肿瘤坏死因子α、Fas水平均呈正相关。     

Intracranial granulocytic sarcoma in a patient with acute myeloid leukemia]

Granulocytic sarcoma (GS) is extramedullary tumor composed of immature leukemic cells. GS is presenting usually as a complication during the course of hematologic neoplasm, such as acute myeloblastic leukemia as well as myeloproliferative and myelodysplastic syndrome. The tumor was also called chroloma based on the green color of the tumorous mass. Central nervous system manifestations of GS are extremely rare. We report a 41-year-old man with acute leukemia type M7, who developed GS in the right occipital lobe after complete remission was achieved. Operative findings revealed the color of the hard tumor was greenish, which suggested the tumor was chroloma. Histological findings showed the tumor was GS. The majority of reported cases of GS in acute myeloid leukemia were M2 FAB classification and have chromosome translocation. Our patient was M7 FAB classification, not have specific chromosome translocation. GS occurrence in AML: M7 patient was extremely rare. This is the first case report of AML: M7 with GS in the central nervous system.     

Meningeal involvement in leukemias and malignant lymphomas of adults: Incidence, course of disease, and treatment for prevention

As prognosis has improved over the last several years, an increasing incidence of meningeal involvement has been recognized in adult patients with acute leukemias and malignant lymphomas. In 210 patients evaluated retrospectively, the incidence of meningeal disease was 33% for patients with acute lymphocytic leukemia (ALL), 20% for patients with acute myelogenous leukemia (AML), 22% for patients with non-Hodgkin's lymphomas with an unfavorable histology (NHL), 3% for patients with chronic myelogenous leukemia (CML), and 1% for patients with chronic lymphocytic leukemia (CLL). In most patients, meningeal involvement appeared several months after diagnosis of acute leukemia, often preceding systemic relapse if bone marrow remission had been achieved before. Prophylactic treatment of the CNS was begun in eight patients with ALL or AML after bone marrow remission was achieved. Of these patients, three with ALL and one with AML were free of disease up to 2 years after diagnosis. Methods, benefits, and risks of prophylactic treatment of the CNS for adult patients are discussed in detail.     

Blast cell-surface and plasma soluble urokinase receptor in acute leukemia patients: relationship to classification and response to therapy.

Plasminogen activation in leukemia has been less well characterized than in other malignancies. However, the increased tendency to bleeding and tissue infiltration by leukemic cells are processes in which plasminogen activation may be involved. We have examined plasma and the peripheral blood mononuclear cell fraction from 80 patients including 53 patients with newly diagnosed acute leukemia and 27 patients with other hematological disorders as well as 21 healthy controls. In 28 of 29 examined patients with acute myeloid leukemia (AML) and in two of three patients with hybrid leukemia we found urokinase receptor (uPAR) on the cell surface, while most (7/9) samples from patients with acute lymphoblastic leukemia (ALL) were negative for uPAR. The plasma mean value for soluble uPAR (suPAR) was significantly elevated in patients with AML and ALL. In AML the highest values were found in patients who had residual disease after several cycles of chemotherapy. Compared to controls the uPA antigen levels in patient plasmas were elevated and decreased along with uPAR during treatment. Our results suggest that cell surface uPAR may be a useful marker for leukemia classification and in our material a high level of plasma suPAR correlated with resistance to chemotherapy in AML.     

Quantitative estimation of elastase-alpha 1-proteinase inhibitor (E-alpha 1 PI) complex in leukemia: marked elevation in cases of acute promyelocytic leukemia

The concentrations of elastase-alpha 1-proteinase inhibitor (E-alpha 1 PI) complex were assayed in 43 patients with various types of leukemia. Marked to moderate elevation of E-alpha 1 PI complex levels was observed in patients with acute myelocytic leukemia (AML), acute promyelocytic leukemia (APL), acute myelomonocytic leukemia (AMMoL), or chronic myelocytic leukemia (CML) at diagnosis. The ratio of E-alpha 1 PI complex concentrations in plasma to leukocyte counts markedly elevated in the patients with APL, especially. During the course of remission induction therapy, levels of E-alpha 1 PI complex decreased in parallel with decline of leukocyte counts in the patients with leukemia other than APL, however the E-alpha 1 PI complex was persistently elevated regardless of leukopenia in some patients with APL. In APL, concentrations of fibrin/fibrinogen degradation products (FDP) markedly increased even when levels of plasmin-alpha 2-antiplasmin complex were within normal limits. However, levels of E-alpha 1 PI complex usually increased in these cases. From these results, it is strongly suggested that promyelocytes contain markedly elevated amounts of elastase which participates in degradation of fibrin or fibrinogen in some cases of APL.     

Low back pain and lumbar radiculopathy as harbingers of acute myeloid leukemia recurrence in a patient with myeloid sarcoma

Myeloid sarcoma (MS) is an extra-osseous, solid collection of myeloblasts. It is associated with myeloid leukemias, and rarely affects the spine. The most common clinical presentation of MS in spine patients is some form of pain related to compression of neural elements. Given that MS is rare, and its imaging characteristics are similar to other more common diagnoses, it is frequently missed on initial presentation. We present a 28-year-old female, in her fifth year of remission from AML, with low back pain and right lumbar radiculopathy. Initially, the leading diagnosis was schwannoma in preference to neurofibroma; however, intra-operative pathology and subsequent bone marrow biopsy revealed the tumor to be MS. This report highlights the difficulties of diagnosis of MS in patients in remission from acute myeloid leukemia. Thus, in patients with a history of leukemia, MS should be considered in the differential diagnosis of any epidural or nerve root tumor. Timely diagnosis and treatment are key to optimal outcomes.     

急性淋巴细胞白血病患者骨髓单个核细胞TAp63基因的表达

背景：据作者查新检索,国内外有关急性白血病患者骨髓单个核细胞TAp63基因表达的报道罕见。 目的：观察急性淋巴细胞白血病患者骨髓单个核细胞TAp63基因的表达及其意义。 方法：50例急性淋巴细胞白血病患者,其中32例急性B淋巴细胞白血病,18例急性T淋巴细胞白血病。同期选择27例非恶性血液病患者作为对照。取肝素抗凝的骨髓液2~4 mL,用Ficoll液分离骨髓单个核细胞,半定量反转录聚合酶链反应法检测TAp63的表达。 结果与结论：50例急性淋巴细胞白血病患者有49例表达TAp63,急性淋巴细胞白血病组明显高于非恶性血液病组(P < 0.05),急性B淋巴细胞白血病表达水平显著高于急性T淋巴细胞白血病(P < 0.05)。动态观察了5例初治急性淋巴细胞白血病化疗后不同阶段TAp63的表达变化,发现初治时TAp63表达,缓解后低表达或不表达,复发后又表达。结果表明TAp63在急性淋巴细胞白血病患者骨髓单个核细胞的表达明显高于非恶性血液病患者,尤其在急性B淋巴细胞白血病患者中高表达。