大剂量甲氨喋呤治疗成人急性淋巴细胞白血病

目的观察大剂量甲氨喋呤(MTX)联合柔红霉素、长春新碱、环磷酰胺、强的松治疗成人急性淋巴细胞性白血病的效果及毒副作用。方法观察12例成人急性淋巴细胞性白血病患者,给予MTX3g/m2联合柔红霉素、长春新碱、环磷酰胺、强的松化疗,同时给予水化、碱化、止吐、保肝、补充能量和液体、生理盐水清洁口腔、防止感染等支持治疗。结果完全缓解(CR)率58.3%,部分缓解(PR)率25.0%,总有效率83.3%。6例中枢神经系统白血病患者5例脑脊液缓解,缓解率83.3%;毒副作用较小。结论大剂量甲氨喋呤联合柔红霉素、长春新碱、环磷酰胺、强的松治疗成人急性淋巴细胞性白血病是一种安全有效的方案。     

Blast cell methotrexate-polyglutamate accumulation in vivo differs by lineage, ploidy, and methotrexate dose in acute lymphoblastic leukemia.

High-dose methotrexate (HDMTX) is a component of most treatment protocols for childhood acute lymphoblastic leukemia (ALL), yet recent studies of receptor-mediated transport and saturable polyglutamylation have questioned its rationale. To investigate this in vivo, methotrexate and its active polyglutamated metabolites (MTX-PG) were measured in bone marrow blasts obtained from 101 children randomized to single-agent therapy with either HDMTX (1 g/m2 per 24 h i.v., n = 47) or low-dose MTX (LDMTX, 30 mg/m2 by mouth every 6 h x 6, n = 54), before remission induction therapy. Blast concentrations of total MTX-PGs (median 460 vs 1380 pmol/10(9) cells) and of long-chain MTX-glu4-6 were both significantly higher after HDMTX (P < 0.001). With either treatment, MTX-PGs were significantly higher in B-lineage blasts than in T-lineage blasts (LDMTX P = 0.001, HDMTX P = 0.03). In a multiple regression analysis of B-lineage ALL, blast MTX-PG was significantly related to MTX dose (or plasma MTX concentration), lymphoblast ploidy (hyperdiploid > nonhyperdiploid), and percentage S-phase. This is the first evidence that HDMTX achieves higher MTX-PG concentrations in ALL blasts in vivo, establishing a rationale for HDMTX in the treatment of childhood ALL, especially T-lineage or nonhyperdiploid B-lineage ALL, disease characteristics associated with a poor prognosis on conventional therapy.     

Evolution of acute lymphoblastic leukemia in mice treated with carrier-bound methotrexate and levamisole

The cytotoxic and antitumoral activities of free or bound to bovine serum albumin (BSA) methotrexate (MTX) and the influence of levamisole (LMS) on these were assayed on murine leukemia. Whereas the in vitro cytotoxic action of MTX was reduced by its conjugation to BSA, in vivo a single dose of 15 mg/kg MTX, which lacked therapeutic effect on tumor-bearing mice, increased the mean survival time (MST) of the animals when given as MTX-BSA. Levamisole slightly increased the MST of the tumor-bearing animals when administered as a 10 mg/kg single dose 7 days after the tumor inoculation. We were unable to achieve synergism between LMS and MTX-BSA when measuring MST and the tumor growth evolution.     

急性淋巴细胞白血病患儿亚甲基四氢叶酸还原酶基因多态性与甲氨蝶呤动态血药浓度相关性的研究

目的本研究旨在探讨亚甲基四氢叶酸还原酶(MTHFR)C677T、A1298C基因多态性对急性淋巴细胞白血病(ALL)患儿使用大剂量甲氨蝶呤(HD-MTX)化疗期间的MTX动态血药浓度的相关关系。方法 35例ALL患儿外周血,提取基因DNA,应用PCR-RFLP方法检测MTHFR C677T、A1298C基因型;应用荧光偏振免疫分析法(FPIA)24h、48h、72h监测患儿外周血中甲氨蝶呤动态血药浓度。结果 MTHFR C677T各基因型间24 h MTX浓度有差异(P0.05),携带CT型者明显高于携带CC型和TT型者;MTHFR C677T各基因型48 h、72 h的MTX浓度未见差异。MTHFR A1298C各基因型24 h、48 h7、2 h的MTX浓度未见差异(P0.05)。结论 MTHFR C677T基因多态性影响ALL患儿HD-MTX化疗期间MTX血药浓度,提示在HD-MTX治疗时可根据检测MTHFR C677T基因型进行个体化治疗。     

儿童急性淋巴细胞白血病患者p73基因异常表达的研究

目的 探讨儿童急性淋巴细胞因病（ALL）发生及发展的机制。方法 应用RT－PCR技术检测61例ALL细胞系和53例儿童ALL患者的p73基因mRNA表达情况，并结合患者的临床资料进行分析。应用DNA甲基化酶谱测定、亚硫酸氢钠修饰的PCR和PCR产物测序等技术对61例ALL细胞系p73基因第1外显子的甲基化进行检测。结果 61个ALL细胞系中p73mRNA阴性表达率为31．1％（61个中19个）；53例原发性儿童ALL患者中，p73 mRNA阴性表达率为26．4％（53例中14例），p73 mRNA阴性表达与ALL无病生存期、总生存期的降低有明显关系。61个ALL细胞系中，39．3％存在p73基因的高甲基化。正常淋巴细胞和不存在p73基因高甲基化的细胞系表达p73 mRNA，大多数高甲基化的细胞系不表达p73 mRNA。结论 儿童ALL患者中p73 mRNA存在较高的阴性表达，其主要机制为p73基因高甲基化，p73基因失活在ALL的发生及发展中起重要作用，p73 mRNA检测对判断儿童ALL患者的预后有一定临床意义。     

临床护理路径在急性淋巴细胞白血病患儿首次大剂量甲氨蝶呤化疗中的应用

目的 探析临床护理路径(CNP)模式在急性淋巴细胞白血病(ALL)患儿首次大剂量甲氨蝶呤化疗护理中的应用效果.方法 选取2018年8月至2020年1月中国科学技术大学附属第一医院收治的106例首次大剂量甲氨蝶呤化疗的ALL患儿作为研究对象,基于随机数字表分成干预组和对照组,各53例.化疗期间对照组采用常规护理模式,研究...     

大剂量甲氨喋呤在儿童急性淋巴细胞白血病中应用观察

目的：观察大剂量甲氨喋呤化疗在儿童急性淋巴细胞白血病中的应用，了解其不良反应。方法：缓解期急性淋巴细胞白血病患儿17例；高危13例，标危4例。累计应用100例次大剂量甲氨喋吟治疗，观察并总结其不良反应发生的种类、发生率、相应处理措施及转归。结果：胃肠道反应、肝功能损害、口腔溃疡、骨髓轻度抑制是大剂量甲氨喋岭常见的不良反应，黏膜炎和重度骨髓抑制形成恶性循环，严重时可致死。结论：充分水化碱化，适宜四氢叶酸钙解救，密切观察病情变化随时处理及必要的心理护理是保证大剂量甲氨喋岭方案顺利进行的关键因素。     

大剂量甲氨蝶呤治疗儿童急性淋巴细胞白血病排泄延迟分析

目的观察儿童急性淋巴细胞白血病(ALL)大剂量甲氨蝶呤(HDMTX)疗法MTX排泄延迟的发生率和不良反应,探讨MTX排泄延迟与MTX剂量、用药方式的关系,及如何减少排泄延迟的发生。方法分析121例497例次儿童ALLHDMTX化疗的临床资料,比较有和无排泄延迟情况下HDMTX的不良反应。并按剂量(3g组和5g组)和用药持续时间(7h组和24h组)分组,比较排泄延迟的发生率、不良反应及各组四氢叶酸钙(CF)的解救剂量。结果总体排泄延迟发生率为12.1%,发生1次排泄延迟的相对概率是30.6%,再次发生的相对概率为45.9%,明显增加(P<0.01)。有排泄延迟的患儿排泄延迟时血小板较无排泄延迟时明显减低(P<0.01),CF解救剂量明显增加(P<0.01)。3g组排泄延迟时口腔黏膜损害较无排泄延迟时更明显(P<0.05),下一疗程化疗延迟(中位延迟4d)。3g组排泄延迟发生率(12.1%)与5g组(12.0%)比较差异无统计学意义(P>0.05),7h用药组排泄延迟发生率(13.6%)与24h用药组(11.9%)比较差异也无统计学意义(P>0.05)。无排泄延迟时5g组仅胃肠道反应较3g组明显增加(P<0.01),而CF解救剂量明显低于3g组(P<0.01)。结论HDMTX排泄延迟情况下骨髓抑制和口腔黏膜损害的不良反应增加,下一疗程化疗延迟,CF用量增加。HDMTX排泄延迟在3～5g/m2剂量范围里与MTX剂量、用药方式无关,而与     

Prevention of neuroleukemia by intrathecal administration of cytosar and methotrexate in acute lymphoblastic leukemia in adults

AIM: To find out whether efficacy of neuroleukemia (NL) prevention by intrathecal administration of cytosar and methotrexate in remission induction phase in adult patients with acute lymphoblastic leukemia (ALL) depends on the risk factors. MATERIALS AND METHODS: The study covered 68 ALL patients. The diagnosis was made by cytological, histological and cytochemical tests of the peripheral blood and bone marrow. Immunophenotyping was performed in 48 patients. The treatment followed the German protocol 04.89 in modification of the Hematological Research Center of the Russian Academy of Medical Sciences. Prevention of NL consisted in intrathecal administration of cytosar (30 mg), methotrexate (15 mg) and dexamethasone (4 mg) once a week for 6 weeks beginning on induction day 1, further in consolidation, reinduction and once in 3 months in maintenance. Radiation of the brain was not conducted. Treatment of leuroleukemia consisted of intrathecal administration of the above drugs twice a week up to normalization of the liquor with subsequent their administration 5 times and craniospinal radiation in a dose 36 Gy. Further intrathecal administrations were made according to the protocol. RESULTS: Correlation was not found between age of the patients and frequency of neuroleukemia onset, between neuroleukemia incidence and peripheral blood leukocytosis at diagnosis. Results of NL prevention with cytosar and methotrexate given intrathecally in induction of remission (14.6% of neurorecurrences) are comparable with the results of NL prevention by radiation of the brain with intrathecal administration of methotrexate obtained in the German cooperative trial. CONCLUSION: NL prevention in ALL adult patients by intrathecal cytosar and methotrexate in remission induction is effective.     

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.     

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2～4mL,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。     

儿童急性淋巴细胞白血病p16基因甲基化及表达水平临床意义

目的 探讨p16基因甲基化及表达水平改变在儿童急性淋巴细胞白血病(ALL)发病机制中的作用.方法 ALL患儿76例,其中初发(uALL)患儿69例、复发(rALL)患儿7例,健康儿童28例纳入对照组.采用荧光定量聚合酶链反应(RTPCR)检测外周血淋巴细胞p16基因mRNA表达;采用基于SYBR Green的甲基化特异性定量PCR(MethySYBR PCR)检测p16基因启动子甲基化水平.结果 ALL患儿淋巴细胞p16基因启动子甲基化水平高于健康儿童,uALL患儿p16基因甲基化水平低于rALL患儿(P＜0.05);ALL患儿p16基因mRNA水平低于健康儿童,uALL患儿p16基因转录水平高于rALL患儿(P＜0.05);p16基因转录水平与其启动子甲基化水平呈负相关关系(r=-0.63,P＜0.05);首次化疗即获完缓解ALL患儿p16基因甲基化水平低于未完全缓解患儿,而p16 mRNA表达水平高于后者(P＜0.05);p16基因低甲基化或高表达水平ALL患儿巩固强化治疗平均缓解时间明显低于p16基因高甲基化或低表达患儿(P＜0.05).结论 p16基因甲基化及表达低下可能与ALL发病有关,p16基因甲基化状态及表达水平检测或可用于儿童ALL预后评估.     

Antagonism by methotrexate on mercaptopurine disposition in lymphoblasts during up-front treatment of acute lymphoblastic leukemia

BACKGROUND: Methotrexate is postulated to enhance mercaptopurine activation to thioguanine (INN, tioguanine) nucleotides, but the interaction has never been studied in vivo in cancer cells. METHODS: We investigated the effect of methotrexate on mercaptopurine disposition in plasma and leukemic blasts during up-front treatment of 233 children with newly diagnosed acute lymphoblastic leukemia. Children were randomized to receive intravenous mercaptopurine (1 g/m(2) over a 6-hour period) or to receive methotrexate (low dose, 6 oral doses of 30 mg/m(2), or high dose, 1 g/m(2) intravenously), followed by intravenous mercaptopurine. All combinations have been previously used in frontline trials for acute lymphoblastic leukemia. RESULTS: Compared with mercaptopurine alone, methotrexate resulted in higher plasma mercaptopurine concentrations (30.3 +/- 14.7 micromol/L versus 23.5 +/- 18.0 micromol/L, P <.001) but, conversely, a 13-fold lower thioguanine nucleotide concentration (0.57 +/- 0.66 pmol/5 x 10(6) cells versus 7.4 +/- 15.2 pmol/5 x 10(6) cells, P <.001) in bone marrow leukemic lymphoblasts. Methotrexate was also associated with higher plasma hypoxanthine concentrations compared with those of patients given mercaptopurine alone (8.7 +/- 13.5 micromol/L versus 3.8 +/- 2.5 micromol/L, P =.029). The percentage change in leukocyte counts measured over a 3-day period showed that mercaptopurine alone had little effect (mean decrease, 20% +/- 33%). In contrast, despite causing lower intracellular thiopurine active metabolite concentrations, methotrexate produced a greater decrease in leukocyte counts (mean, 53% +/- 35%) compared with those in patients receiving mercaptopurine alone (P <.0001). CONCLUSION: These pharmacologic findings in the target tissue are consistent with the recently demonstrated lack of clinical benefit of intravenous mercaptopurine in combination with methotrexate. We conclude that, in the setting of newly diagnosed acute lymphoblastic leukemia, methotrexate antagonizes thiopurine metabolite disposition in leukemic blasts after intravenous mercaptopurine.     

左旋门冬酰胺酶治疗儿童急性淋巴细胞白血病的药代和药效动力学研究

目的观察急性淋巴细胞白血病(ALL)患儿应用常规剂量左旋门冬酰胺酶(L—asp)后，其血浆中L—asp活性变化及底物门冬酰胺(ASN)耗竭的规律，寻找L—asp合理的用药方法。方法ALL患儿15例(340份外周血样本)，基本联合化疗采用VDLP(D)方案，诱导缓解用VDLP方案，早期强化及定期维持治疗阶段用VDLD方案，其国L-asp每次10000U／m^2，静脉滴注，隔天1次，共10次，分别检测应用常规剂量L—asp(每次10000U／m^2)疗程前、疗程中及疗程后多个时间点患儿血浆L—asp活性及ASN浓度。结果在用药过程中，血浆L—asp活性峰值有逐渐增高的趋势，在第8次用药后达到最高峰，之后缓慢下降，而血浆中ASN浓度始终维持在极低的水平，处于“几乎完全缺乏”甚至“完全缺乏”的状态，停药后，血浆中L-asp活性持续7d维持100U／L以上，ASN浓度持续7d保持在“几乎完全缺乏”水平，随后逐渐上升至用药前水平结论目前常规用药剂量的L—asp能够完全耗竭血浆ASN，由于L—asp应用后达到药物效应所需的血浆活性在停药后可持续7d，且L—asp相关并发症与其血浆中活性相关，为减少并发症的发生，目前L-asp的用量及用药间隔时间需要更深入探讨，使L-asp的使用更趋合理。     

Activation of the alternative pathway of complement in childhood acute lymphoblastic leukemia.

Sequential studies in children with acute lymphoblastic leukemia have demonstrated that at diagnosis or relapse there is defective utilization of complement by the alternative pathway. Thus, the sera of 17/18 patients fail to completely consume C3 to C9 when incubated with zymosan or cobra venom factor (CoF). This underutilization is due to a specific inhibitor of C3 activation which has been partially isolated. By remission, the inhibotor disappears and the CoF and zymosan assays return to normal. In addition, serum levels of C3 and factor B are elevated at the time of diagnosis or relapse but fall to below 3 S.D. from the mean in nearly 60 per cent of the cases during induction therapy. Similarly, serum C4 levels which are normal at diagnosis fall to less that 3 S.D. from the mean in 7/12 cases during treatment. Low C3 levels correlate well with factor B values, suggesting that if C3 to C9 are utilized after the inhibitor has been eliminated, such utilization occurs primarily through the alternative pathway. Presumably, as illustrated by the low C4 levels, this activity is mediated by the ampliciation loop of the alternative pathway involving classical pathway generation of C3b.     

As_2O_3诱导急性淋巴细胞白血病细胞系Molt4细胞p15基因表达的研究

目的　探讨砷剂与甲基化的关系。方法　先用甲基化特异的PCR检测急性T淋巴细胞白血病细胞系Molt4细胞p15基因甲基化 ,然后用RT PCR方法检测用As2 O3 处理或未处理Molt4细胞的p15基因mRNA表达 ,用流式细胞仪检测细胞周期并绘制生长曲线。结果　Molt4细胞p15基因发生甲基化 ,p15mRNA不表达 ,Molt4细胞与As2 O3 一起培养后 ,p15基因mRNA重新表达 ,G0 G1 期细胞明显增多 ,Molt4细胞生长受到抑制。结论　As2 O3 能够去p15基因甲基化 ,激活p15基因的mRNA表达 ,恢复p15基因的细胞周期负调节功能。     

As2O3诱导急性淋巴细胞白血病细胞系Molt4细胞p15基因表达的研究

目的 探讨砷剂与甲基化的关系。方法 先用甲基化特异的PCR检测急性T淋巴细胞白血病细胞系Molt4细胞p15基因甲基化,然后用RT－PCR方法检测用As2O3处理或未处理Molt4细胞的p15基因mRNA表达,用流式细胞仪检测细胞周期并绘制生长曲线。结果 Molt4细胞p15基因发生甲基化,p15 mRNA不表达,Molt4细胞与As2O3一起培养后,p15基因mRNA重新表达,G0－G1期细胞明显增多,Molt4细胞生长受到抑制。结论 As2O3能够去p15基因甲基化,激活p15基因的mRNA表达,恢复p15基因的细胞周期负调节功能。