流式细胞仪分析儿童急性淋巴细胞白血病免疫表型63例

为了了解儿童急性淋巴细胞白血病的免疫表型,应用一组单克隆体及流式细胞仪间接免疫荧光法对６３例初治儿童急性淋巴细胞白血病（ＡＬＬ）免疫分型,Ｔ淋巴细胞、Ｂ淋巴细胞型、Ｔ／Ｂ混合型及裸核ＡＬＬ分别为４１．３％、４６％、９．５％、３．２％,髓系抗原（Ｍｙ）及ＣＤ３４表达率为１７．４％和２８．５％,Ｍｙ＾＋ＡＬＬ中ＣＤ３４表达为５４．５％,显著高于ＭｙＡＬＬ（２３．３％）,Ｔ／Ｂ混合型ＡＬＬ对Ｍｙ及ＣＤ３     

急性白血病细胞凋亡、DNA含量的流式分析

目的分析急性白血病(急淋初治37例、缓解期32例，急非淋9例)细胞凋亡量、DNA含量的变化及临床意义。方法采用流式细胞术(FCM)碘化丙啶插入性DNA定量染色法。结果凋亡百分率在缓解期最高，为19.92±8.05，初治组为8.16±3.12，与正常儿有显著性差异；白血病儿DNA指数(DI值)、异倍体(AN)率、SPF值(S期细胞百分比)均显著性增高，异倍体的发生与肝脾大小、白细胞数有关，与FAB分型无关。结论小儿白血病细胞存在DNA含量的异常，凋亡量、SPF等指标对提示预后有一定的参考价值。     

儿童急性淋巴细胞性白血病细胞中细胞周期相关因子Cdt1的表达

目的 研究细胞周期相关因子Cdt1在儿童急性淋巴细胞性白血病(ALL)细胞中的表达.方法 以15例首次确诊的儿童ALL为实验组,以10例同期住院非肿瘤患者作为对照组,应用流式细胞术检测实验组治疗前、后及对照组细胞中Cdt1表达水平,并进行比较.结果 儿童ALL治疗后(3.56±2.43)与治疗前(4.12 ±2.47)比较Cdt1表达明显下降(P<0.01);儿童ALL治疗前Cdt1表达显著高于对照组(2.33 ±1.32,P<0.05);儿童ALL治疗后与对照组比较Cdt1表达差异无统计学意义(P>0.05).结论 Cdt1可能参与儿童ALL的发生发展.可以通过监测ALL治疗前、后细胞中Cdt1表达水平,判断疗效. [临床儿科杂志,2009,27(10):955-956]     

急性淋巴细胞白血病患儿体内肿瘤细胞的凋亡及其临床意义

目的 检测急性淋巴细胞白血病（ALL）患儿体内肿瘤细胞的凋亡情况,并初步探讨其临床意义。方法 用Annexin-V-FITC/PI双标记法检测22例初治ALL患儿化疗前后骨髓白血病细胞的凋亡情况并与患儿的近期化疗疗效及DNA倍性、免疫表型等临床特征进行相关分析。结果 化疗前无一例检出凋亡细胞,化疗后13／22例凋亡检测阳性。化疗后患儿白血病细胞的凋亡情况与其早期缓解、DNA倍性、免疫表型有关（P均＜0．05）,与WBC数、年龄、性别、髓外浸润情况无关（P均＞0．05）。结论 Annexin-V-FITC/PI双标记法是一种简单、敏感的检测体内凋亡细胞的方法;化疗后体内白血病细胞的凋亡情况与近期化疗疗效一致;DNA指数1．16－1．6的ALL患儿预后好,与其白血病细胞易于凋亡有关;成熟B－ALL、T－ALL患儿预后差,与其白血病细胞不易凋亡有关。     

儿童急性淋巴细胞白血病细胞周期分布及临床预后的研究

目的 研究儿童急性淋巴细胞白血病（ALL）骨髓细胞DNA含量和倍性情况,了解其生物学意义及与临床预后的关系。方法 取29例初诊ALL儿及10例骨髓像正常的非白血病患儿的骨髓细胞,采用流式细胞术测定,获取各标本细胞周期时相百分比。本组资料平均随访48个月。结果 29例初诊ALL儿检出8例为超二倍体,对照组无异倍体检出。ALL儿的G0／G1％明显高于对照组（P＜0.01）,S％和S＋G2／M％则明显低于对照组（P＜0.01）。经随访发现,初诊DI＞1.08的患儿死亡率为225,DI＜1.08的患儿死亡率为40％,但统计学显著性检验无明显差异（P＝0.311）。初诊G2／M％＞4.0的患儿死亡率为12.5％,G2／M％＜4.0的患儿死亡率为42.7％,但统计学显著性检查无明显差异（P＝0.135）。结论 儿童ALL儿DNA测定对临床预后有一定指导作用。     

流式细胞仪分析儿童急性淋巴细胞白血病免疫表型63例

为了了解儿童急性淋巴细胞白血病的免疫表型,应用一组单克隆抗体及流式细胞仪间接免疫荧光法对６３例初治儿童急性淋巴细胞白血病(ＡＬＬ）免疫分型：Ｔ淋巴细胞型、Ｂ淋巴细胞型、Ｔ／Ｂ混合型及裸核（Ｎｕｌｌ型）ＡＬＬ分别为４１．３％、４６％、９．５％、３．２％,髓系抗原（Ｍｙ）及ＣＤ(３４)表达率为１７．４％和２８．５％,Ｍｙ＋ＡＬＬ中ＣＤ(３４)表达为５４．５％,显著高于Ｍｙ－ＡＬＬ（２３．３％）（Ｐ＜０．０１）,Ｔ／Ｂ混合型ＡＬＬ对Ｍｙ及ＣＤ(３４)均有高表达,与Ｔ、Ｂ－ＡＬＬ相比均有显著差异（Ｐ＜０．０１－０．０５）     

儿童急性淋巴细胞白血病复发与基因突变相关性

急性淋巴细胞白血病是儿童恶性肿瘤中最常见的类型,急性淋巴细胞白血病复发仍然是治疗的难题.随着近几年关于儿童急性淋巴细胞白血病复发机制的研究逐渐深入,越来越多的基因异常已经被证实与儿童急性淋巴细胞白血病复发相关,包括IKZF1缺失、PRED1缺失、JAK突变、CREBBP突变、CEBPE突变、ARID5B突变等.该文重点综述以上基因突变对儿童急性淋巴细胞白血病复发的影响.     

T型急性淋巴细胞白血病患儿的免疫表型分析

目的　应用流式细胞术分析 17例儿童T型急性淋巴细胞白血病 (T ALL)的免疫表型特点。方法　用三色荧光标记法置于流式细胞仪检测 17例T ALL ,并随机抽取B ALL 17例进行比较分析。结果　T ALL占急性淋巴细胞白血病 13% ,其髓系抗原表达占 2 3.5 % ,B系抗原表达占 4 1.0 % ;CD2表达率显著低于CD5、CD7、cyCD3(P <0 .0 0 1) ;在CD45散点图中 ,T ALL幼稚细胞群在CD45(y轴 )均值明显高于B ALL(P <0 .0 5 )。结论　T ALL时CD5、CD7、cyCD3特异性高 ;在CD4 5 vsSSC散点图中T ALL分布特性亦不同于B ALL。     

细胞周期相关因子geminin在儿童急性淋巴细胞白血病中的表达

目的 研究细胞周期相关因子geminin蛋白及其mRNA在儿童急性淋巴细胞白血病中的表达.方法 以16例首次诊断且未治疗的急性淋巴细胞白血病患儿为实验组,16例正常儿童为对照组,采集其静脉血进行淋巴细胞分离,应用免疫组织化学染色法检测geminin蛋白在两组中的表达;用RT-PCR法检测gemininmRNA在两组中的表达.使用SPSS软件进行数据分析.结果 实验组geminin蛋白及geminin mRNA表达均高于对照组(P均<0.05);对照组geminin蛋白有表达,但其相应的mRNA不表达.结论 急性淋巴细胞性白血病细胞geminin过度表达,可能参与儿童急性淋巴细胞白血病的发生发展;正常淋巴细胞geminin存在蛋白与其相应的mRNA表达不一致的现象.     

儿童急性淋巴细胞白血病及阿霉素所致Jurkat细胞凋亡时WAVE1表达研究

目的：初步探讨WASP家族Verprolin同源蛋白1（WAVE1）在急性淋巴细胞白血病（ALL）发病中可能的作用及意义。方法：运用逆转录-聚合酶链反应（RT-PCR）半定量法和蛋白质印迹（Western blotting）分别检测40例ALL初治患儿、15例化疗完全缓解半年、4例复发、10例非白血病患儿BMMCs中WAVE1 mRNA和蛋白的表达水平。以不同浓度阿霉素处理Jurkat细胞：①采用MTT法测细胞增殖;②采用流式细胞仪检测细胞凋亡率;③采用RT-PCR和Western blotting检测WAVE1表达。结果：ALL初治和复发患儿BMMCs均有WAVE1 mRNA和蛋白的表达,对照和缓解组BMMCs中mRNA和蛋白低表达或无表达,初治和复发组的WAVE1 mRNA和蛋白的表达表达水平显著高于化疗后缓解组和对照组（P<0.01）。阿霉素明显抑制Jurkat细胞增殖,抑制作用呈剂量时间依赖效应（P<0.05）;阿霉素作用24 h后,细胞凋亡随药物浓度增增高而增加,与对照组相比,差异有显著性(P<0.05);细胞WAVE1 mRNA和蛋白表达水平随阿霉素处理浓度的增高而降低,与对照组比较,差异有显著性(P<0.05)。结论：WAVE1在ALL患儿BMMCs中高表达;WAVE1可能与儿童ALL病程相关,它可能成为动态检测儿童ALL病情的一个新指标。     

儿童急性淋巴细胞白血病微小残留病与复发的相关分析

目的 分析小儿急性淋巴细胞白血病(急淋)缓解时和缓解不同时期微小残留病(MRD)的水平与复发的相关性。方法：用极限稀释定量PCR法和巢式PCR法追踪检测MRD，数据处理用Kaplan Meier方法及COX回归模型等。结果：46例急淋患儿缓解时MRD值与骨髓复发呈正相关(r=0.4396，P<0.01)，骨髓复发组MRD值为7.359×10-3，与未复发组MRD值(3.954×10-4)差异有显著性(P<0.01)；缓解期MRD持续阳性或由阴性转为阳性者，骨髓复发的相对危险度明显增高(P<0.05)。结论：缓解时MRD水平及缓解期MRD定性结果可作为估计急淋预后的一个重要指标，动态追踪检测MRD是指导治疗和预防复发的有效手段。     

两种不同方案治疗儿童急性淋巴细胞白血病疗效分析

目的：比较两种治疗方案对小儿急性淋巴细胞白血病(ALL)治疗的近期完全缓解率(CR)与持续完全缓解(CCR),探讨影响小儿ALL长期生存的有关因素。方法：根据化疗方案所用药物不同将44例患儿分为2组：一般化疗组(A组),诱导治疗采用VCP(长春新碱、环磷酰胺、强的松),庇护所预防使用二联鞘注(甲氨喋呤、地塞米松),维持治疗使用6 巯嘌呤与甲氨喋呤,加强强化用VCP及COAP(长春新碱、环磷酰胺、阿糖胞苷、强的松)交替;强烈化疗组(B组),按照1993年广西北海会议制定的小儿急性白血病诊疗建议进行序贯治疗,其中大剂量甲氨喋呤(HD-MTX)采用每次 1.5～2.0 g/m2及三联鞘注(甲氨喋呤、阿糖胞苷、地塞米松)。结果：两种方案经过4周治疗44例患儿均获CR,但达到CR的时间A组为(3.83±0.41)周,长于B组(3.00±0.82)周(P0.05)。结论：强烈化疗组不仅近期完全缓解所用时间短,而且在CCR及预防疾病复发上明显优于一般化疗组,虽然强烈化疗后合并各种感染的机会增多,但只要积极采取相应的预防措施,取得家长的密切配合可以使其发生率降低。     

Immunological Classification of Childhood Acute Lymphoblastic Leukemia

Seven hundred and forty-four newly diagnosed patients with acute leukemias between 1978 and 1990 were classified on the basis of immunological phenotypes. The majority of the patients were enrolled in the Tokyo Children's Cancer Study Group (TCCSG) studies. The incidence of subclassification of acute leukemias in this study was as follows: 522 patients with ALL (70%), 139 patients with ANLL (18%), 29 patients with biphenotypic leukemia, 8 patients with Ph1-positive acute leukemia (Ph^-AL), and 45 patients with infant leukemia. ALLs were classified into common ALL (cALL, 77%), T-ALL (15%), B-ALL (4%), and unclassified ALL (3%). The incidence of ALL subtypes in this study reflected those of TCCSG. Biphenotypic leukemias were categorized into 4 groups as follows; 1) cALL with positive myelomonocytic antigen(s) (N = 11), 2) unclassified ALL with positive myelomonocytic antigen(s) (N = 5), 3) ANLL with positive B-lymphoid antigen(s) (N = 4), and 4) acute leukemia with positive T-lymphoid and myeloid antigen(s). Infant leukemias were classified into ALL type (N = 27) and ANLL type (N = 18). In this present study, clinical features and immunological phenotypes of the acute leukemias with a poor prognosis, i.e. biphenotypic leukemia, Ph^-AL, and infant leukemia are analyzed and discussed.     

COALL-97方案治疗急性淋巴细胞白血病临床及实验室研究

目的　引用德国儿童急性淋巴细胞白血病协作组 (cooperativeALLstudygroup ,COALL)的COALL 97方案治疗急性淋巴细胞白血病 (ALL) ,从临床疗效防治及目前国内外选用有效治疗ALL的药物 ,左旋门冬酰胺酶 (L Asp)的药代动力学方面 ,探讨在我国治疗儿童ALL的最佳方案及L Asp的最佳给药方式。方法　ALL患儿 1 2例自愿接受COALL 97方案在我院完成早期强化治疗后 ,回当地继续口服巯基嘌呤 (6 MP) /硫鸟嘌呤 (6 TG)加甲氨蝶呤 (MTX)持续治疗 ,每隔 3个月、半年来院复查 ,总疗程 1 .5～ 2 .0年。结果　诱导治疗后d1 4做骨髓检查 8例呈缓解骨髓像 ,治疗 2 8d后 1 2例均完全缓解 (CR) ,CR率1 0 0 % ,HR ALL患儿 1例CR后 1 0个月时骨髓复发并重症感染死亡。应用高压液相色谱技术 (HR HPLC)检测 1次注射 40 0 0 0U/m2 L Asp后能使血清和脑脊液 (CSF)中门冬酰胺 (ASN)浓度降低 ,并持续至第 5周后回升。 结论　COALL 97方案可被我国ALL患儿所接受 ;在治疗ALL的临床实践中 ,该方案确有可借鉴之处 ;1次 / 2～ 6周静脉注射 40 0 0 0U/m2 L Asp联合化疗的用药方式不仅作用强、持续时间长 ,耐药性小、不良反应少 ,且避免患儿每天或隔天静脉注射痛苦及患儿家长的经济和精神负担。     

Prognosis after Relapse in Acute Lymphoblastic Leukemia in Childhood

This is a survey of all the 265 relapses occurring in 515 children with ALL diagnosed in Sweden in the years 1973-1980. Two hundred and nineteen relapses occurred on therapy, and 46 after discontinuation of therapy. Bone marrow was involved in the relapse in 71% and 67% of the two groups, respectively. Only 38/265 (14%) children with relapse were still alive at follow-up in January 1985. Of these, 16/219 (7%) had relapsed during therapy (median survival time after relapse 9 months) compared to 22/46 children (48%) with a relapse after cessation of therapy (median 43 months). The prognosis was better if relapse occurred after cessation of therapy and in children with isolated testicular relapse. Thirteen children were bone marrow transplanted, and 6 of these were alive at follow-up. It is concluded that children with ALL relapse have very bad prognosis with cytostatic regimens used today, especially if the bone marrow is involved.     

Ocular Granulocytic Sarcoma in Childhood Acute Myelogenous Leukemia

We report two children of Greek origin with granulocytic sarcoma of the orbit and acute myelogenous leukemia. In the first case the orbital tumor was the initial manifestation of the disease, while the other occurred during hematological remission.     

Cognitive Function in Long Survivors of Childhood Acute Lymphoblastic Leukemia

Thirty long survivors of childhood acute lymphoblastic leukemia were compared with their healthy siblings on cognitive and neuropsychological measures. The subjects were comparable in treatment variables except for type of central nervous system prophylaxis received. Thirteen were given radiotherapy with intrathecal medication, and seventeen received only intrathecal treatment. Patients receiving only intrathecal medication did not differ from controls. Patients receiving radiotherapy scored lower on several measures including Wechsler Full Scale and Performance IQ. Irradiated girls scored lower than boys on most measures. Children whose clinical management has included radiation therapy appear to be at risk for later mild cognitive deficits. No consistent pattern of neuropsychological deficits has emerged and observed deficit patterns may reflect individual vulnerabilities.     

Birth Weight as a Risk Factor for Childhood Acute Lymphoblastic Leukemia

Increased birth weight previously has been reported to be associated with childhood acute leukemia although the etiologic importance of this finding remains unclear. To further assess birth weight and associated parameters as a risk factor for childhood leukemia, a casel control study was performed using children with acute lymphoblastic leukemia (ALL) born in the state of Minnesota and diagnosed since 1969. Data obtained from birth registrations of 219 cases were compared with two control groups matched on date and county of birth (group I) or year of birth (group II). No significant differences were observed in mean birth weights of cases and controls. Statistically significant associations with birth weights greater than 3800 g were identified in cases diagnosed within the first 4 years of life. No associations were found between birth weight and ALL for case children diagnosed after 4 years of age. Factors that might be associated with increased birth weight, including maternal age, birth order, length of gestation, and socioeconomic status as measured by paternal education, were not found to be associated with an increased risk for ALL. The significance of the finding of high birth weight as a risk factor for childhood ALL remains unknown but suggests that pregnancy-related events may be of importance in the etiology of ALL in young children.