Comparison of microtransplantation,chemotherapy and allogeneic transplantation in post-remission therapy for Philadelphia chromosome-positive acute lymphoblastic leukemia

Patients with Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph⁺ ALL) have poor prognosis, and the efficacy of chemotherapy plus tyrosine kinase inhibitors (TKIs) followed by mismatched donor stem cell infusion (microtransplantation, MST) has not been determined. We retrospectively summarized 45 patients including 11 undergoing MST with TKIs, 17 receiving allogeneic transplant and 17 undergoing chemotherapy with TKIs. Improved 4-year overall survival rate was observed in the MST group (91%) compared with either transplant group (31%, P = .005) or chemotherapy group (36%, P = .013). The MST group also had higher 2-year and 4-year leukemia-free survival rates (91% and 72%, respectively) compared with either transplant group (33%, P = .005 and 33%, P = .021, respectively) or chemotherapy group (41%, P = .017 and 31%, P = .023, respectively). 2-year and 4-year cumulative incidences of hematologic relapse were lower in the MST group (9% and 28%, respectively) compared with those in the chemotherapy group (56%, P = .025 and 67%, P = .034, respectively). In patients undergoing MST, donor microchimerism was detected (1.07 × 10^-5 to 6.6 × 10^-4 copies from 9 to 1499 days) in 7 patients, and donor/patient-derived HLA*0201/2402⁺WT1⁺CD8⁺ T cells were found from 0.05% to 0.67% in 6 patients. MST may provide a favorable treatment for patients with Ph⁺ ALL.     

微移植联合化疗与单纯化疗治疗较高危骨髓增生异常综合征的疗效比较

目的:比较微移植联合地西他滨序贯改良HAG方案与单纯化疗治疗较高危骨髓增生异常综合征(MDS)患者的临床疗效及不良反应，探讨微移植在MDS中应用的可行性和有效性。方法:回顾性分析我院共33例较高危骨髓增生异常综合征患者经微移植联合地西他滨序贯改良HAG方案（21例）和单纯化疗（12例）治疗过程及转归。结果:微移植组14例（66.7%）CR，单纯化疗组3例（25.0%）CR，两组之间差异有统计学意义（P<0.05）。微移植组与单纯化疗组中性粒细胞中位恢复时间分别为13 d、15.5 d，两组之间差异有统计学意义（P<0.05），血小板中位恢复时间分别为16 d、18 d，两组之间差异有统计学意义（P<0.05）。微移植组1年OS率达90.5%，2年OS率达81.0%，单纯化疗组1年OS率达50.0%，2年OS率达41.7%，两组之间差异有统计学意义（P<0.05）。微移植组1年PFS率达57.1%，2年PFS率达42.9%，单纯化疗组1年PFS率达 41.7%，2年PFS率达25.0%。结论:微移植联合化疗治疗较高危骨髓增生异常综合征疗效、安全性均较好。     

微移植治疗老年急性髓性白血病患者的护理

目的分析微移植治疗老年急性髓性白血病患者的护理方法及效果。方法将本院2016年1月-2019年4月的40例老年急性髓性白血病患者,所有患者给予微移植治疗,随机分组,普通护理组对于我院的老年急性髓性白血病患者给予普通护理,全方位护理组对于我院的老年急性髓性白血病患者开展全方位护理。比较两组满意度;患者自觉舒适度评分;护理前后WHOQOL-BREF生活量表生理状况、心理状况、社会关系、环境状况和总体感觉评分;感染发生率。结果全方位护理组的满意度是100.00%(20例),普通护理组则是75.00%(15例),P <0.05。护理前两组WHOQOL-BREF生活量表生理状况、心理状况、社会关系、环境状况和总体感觉评分接近,P> 0.05;护理后全方位护理组WHOQOL-BREF生活量表生理状况、心理状况、社会关系、环境状况和总体感觉评分优于普通护理组,P <0.05;全方位护理组患者自觉舒适度评分均比普通护理组的时间短,P <0.05;全方位护理组感染发生率更少,P <0.05。结论老年急性髓性白血病患者实施全方位护理效果确切,可有效改善老年急性髓性白血病患者的生活质量,提高舒适度,并降低感染发生率,提高满意度。     

Microtransplantation of whole ganglionic eminence cells ameliorates motor deficit,enlarges the volume of grafts,and prolongs survival in a rat model of Huntington's disease

Studies have demonstrated that embryonic cell therapy is a potential approach for the treatment of Huntington's disease (HD). However, because of the limited resource of embryos, greater attention is needed in developing more efficient surgical techniques that not only enhance the therapy outcome but also avoid inefficient therapeutics of transplantation. In this study, we explored the curative effects of two different transplantation methods using a rat model of HD. Whole ganglionic eminence (WGE) cells or phosphate‐buffered saline were transplanted into unilateral striatum of quinolinic acid (QA)‐lesioned rats using microtransplantation instruments (with an outer diameter of 50 μm) or traditional transplantation instruments (with an outer diameter of 470 μm). Apomorphine‐induced rotation test and adjusting step test were assessed after QA‐induced lesion and 2, 4, 6, 8, 10, and 12 weeks after transplantation. The expression of neuronal nuclei (NeuN), dopamine, cAMP‐regulated phosphoprotein of molecular weight 32 kDa (DARPP‐32), and glial fibrillary acidic protein (GFAP) was analyzed at 12 weeks after transplantation. We observed that microtransplanted rats performed better in the stepping test and had higher numbers of DARPP‐32‐positive cells compared with traditionally transplanted rats. Moreover, microtransplantation group showed lower GFAP expression surrounding the grafts in unilateral striatum and a higher survival rate posttransplantation compared with the traditional transplantation group. We conclude that microtransplantation is capable of enhancing therapeutic efficacy in the rat model of HD. This finding establishes the basis of an alternative transplantation strategy for treatment of HD. © 2013 Wiley Periodicals, Inc.     

BDNF modulates GABAA receptors microtransplanted from the human epileptic brain to Xenopus oocytes

Cell membranes isolated from brain tissues, obtained surgically from six patients afflicted with drug-resistant temporal lobe epilepsy and from one nonepileptic patient afflicted with a cerebral oligodendroglioma, were injected into frog oocytes. By using this approach, the oocytes acquire human GABAA receptors, and we have shown previously that the "epileptic receptors" (receptors transplanted from epileptic brains) display a marked run-down during repetitive applications of GABA. It was found that exposure to the neurotrophin BDNF increased the amplitude of the "GABA currents" (currents elicited by GABA) generated by the epileptic receptors and decreased their run-down; both events being blocked by K252A, a neurotrophin tyrosine kinase receptor B inhibitor. These effects of BDNF were not mimicked by nerve growth factor. In contrast, the GABAA receptors transplanted from the nonepileptic human hippocampal uncus (obtained during surgical resection as part of the nontumoral tissue from the oligodendroglioma margins) or receptors expressed by injecting rat recombinant alpha1beta2gamma2 GABAA receptor subunit cDNAs generated GABA currents whose time-course and run-down were not altered by BDNF. Loading the oocytes with the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetate-acetoxymethyl ester (BAPTA-AM), or treating them with Rp-8-Br-cAMP, an inhibitor of the cAMP-dependent PKA, did not alter the GABA currents. However, staurosporine (a broad spectrum PK inhibitor), bisindolylmaleimide I (a PKC inhibitor), and U73122 (a phospholipase C inhibitor) blocked the BDNF-induced effects on the epileptic GABA currents. Our results indicate that BDNF potentiates the epileptic GABAA currents and antagonizes their use-dependent run-down, thus strengthening GABAergic inhibition, probably by means of activation of tyrosine kinase receptor B receptors and of both PLC and PKC.     

白血病小鼠H-2全不相合微移植模型的建立及鉴定

本研究旨在建立白血病小鼠H-2全不相合微移植模型并进行鉴定。受、供鼠分别为雌性BALB／c和雄性C57BL／6J,H-2为全不相合。受鼠微移植前5d静脉接种WEHI-3细胞约1×10^6个,微移植前3d开始给予MA化疗（米托蒽醌＋阿糖胞苷）,0d末次化疗后8h之内回输经G—CSF动员的供体脾单个核细胞,回输细胞数分别为（3、6、12）×10^7个。化疗对照组用等量生理盐水。所有受鼠不予GVHD预防。比较各组早期死亡率、白细胞恢复及白血病负荷情况。RT—PCR法检测微移植后供体嵌合率。从临床表现和病理情况综合评价微移植组小鼠GVHD情况。结果表明,化疗对照组早期死亡率为25％,（3、6、12）×10^7组分别为16．67％、8．33％、8．33％。（3、6）×10^7组白细胞恢复明显优于化疗对照和12×10^7组（P〈0．05）。白血病负荷（3、6、12）×10^7组明显低于化疗对照组（P〈0．01）,而（6、12）×10^7组明显低于3X10^7组（P〈0．05）。微移植后供体成分在2周之内以微嵌合形式存在。微移植组小鼠均未发生明显GVHD证据。结论：成功建立了白血病小鼠H-2全不相合微移植模型,为后续研究及临床微移植相关研究提供了基础。     

Acetylcholine receptors from human muscle as pharmacological targets for ALS therapy

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease affecting motor neurons that leads to progressive paralysis of skeletal muscle. Studies of ALS have revealed defects in expression of acetylcholine receptors (AChRs) in skeletal muscle that occur even in the absence of motor neuron anomalies. The endocannabinoid palmitoylethanolamide (PEA) modified the clinical conditions in one ALS patient, improving muscle force and respiratory efficacy. By microtransplanting muscle membranes from selected ALS patients into Xenopus oocytes, we show that PEA reduces the desensitization of acetylcholine-evoked currents after repetitive neurotransmitter application (i.e., rundown). The same effect was observed using muscle samples from denervated (non-ALS) control patients. The expression of human recombinant α1β1γδ (γ-AChRs) and α1β1εδ AChRs (ε-AChRs) in Xenopus oocytes revealed that PEA selectively affected the rundown of ACh currents in ε-AChRs. A clear up-regulation of the α1 subunit in muscle from ALS patients compared with that from non-ALS patients was found by quantitative PCR, but no differential expression was found for other subunits. Clinically, ALS patients treated with PEA showed a lower decrease in their forced vital capacity (FVC) over time as compared with untreated ALS patients, suggesting that PEA can enhance pulmonary function in ALS. In the present work, data were collected from a cohort of 76 ALS patients and 17 denervated patients. Our results strengthen the evidence for the role of skeletal muscle in ALS pathogenesis and pave the way for the development of new drugs to hamper the clinical effects of the disease.Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease marked by the degeneration of motor neurons. It leads to progressive paralysis ending with the death of the patient 3–5 y after diagnosis. Although most ALS cases are sporadic, about 10% are familial and are linked to monogenic mutations in different genes such as the gene for superoxide dismutase 1 (SOD1), genes encoding RNA-binding proteins such as TARDBP and FUS, or the noncoding region of the poorly characterized C9ORF72 gene (1). These mutations have been used to develop mammalian animal models that mimic some of the symptomatology and to investigate some of the molecular mechanisms of the disease. In mice, as well as in patients, the first observed disease event is the destruction of the neuromuscular junction (NMJ) (2). Studies have uncovered defects in the skeletal muscle that occur even in the absence of motor neuron anomalies, supporting the “dying-back” hypothesis in which distal motor endplate degeneration plays a key role in the progression of the disease (2–4). NMJs of ALS patients consistently show electrophysiological properties distinctly different from those of patients with pure denervation (5, 6). Using muscle biopsies from sporadic ALS patients, we previously characterized the nicotinic acetylcholine currents resulting from the activation of functional acetylcholine receptors (AChRs) (5). Upon denervation, receptors formed by the α1β1γδ (γ-AChR) instead of the α1β1εδ (ε-AChR) subunits appear widely distributed over the entire sarcolemma (7, 8). In ALS patients, in whom muscle denervation and abnormal reinnervation are pathological hallmarks of the disease, both γ- and ε-AChRs are present. In addition, we showed that riluzole, the only approved drug available against ALS, may affect AChR function in ALS muscle (6). These findings strengthen the hypothesis that pathogenic events target the NMJ directly and suggest that its protection could be a useful therapeutic strategy.The cannabinoid system, consisting of cannabinoid (CB) receptors, endocannabinoids (eCBs), and enzymes involved in the synthesis and degradation of these eCBs, has been shown to be a key player in ALS etiology (9, 10). The eCB-related molecule palmitoylethanolamide (PEA) induces anti-inflammatory effects through the PPARα receptor without binding to CB receptors and consequently without side effects (11, 12). Notably, PEA was able to improve pulmonary function and clinical conditions in a single case of ALS (13). Here, taking advantage of the microtransplantation technique by which cell membranes isolated from human muscle are injected into Xenopus oocytes (14), we investigated whether PEA can modulate AChR currents using voltage-clamp intracellular recordings in oocytes transplanted with membranes from ALS muscle. We measured the composition of AChR subunits in ALS muscle compared with non-ALS denervated muscle by quantitative PCR (qPCR) analysis. In addition, we studied the effect of PEA in a cohort of ALS patients to determine whether this compound can improve the clinical characteristics of patients, with particular attention to muscle force and respiratory capacity.     

Adenosine receptor antagonists alter the stability of human epileptic GABAA receptors

We examined how the endogenous anticonvulsant adenosine might influence γ-aminobutyric acid type A (GABA_A) receptor stability and which adenosine receptors (ARs) were involved. Upon repetitive activation (GABA 500 μM), GABA_A receptors, microtransplanted into Xenopus oocytes from neurosurgically resected epileptic human nervous tissues, exhibited an obvious GABA_A-current (I_GABA) run-down, which was consistently and significantly reduced by treatment with the nonselective adenosine receptor antagonist CGS15943 (100 nM) or with adenosine deaminase (ADA) (1 units/ml), that inactivates adenosine. It was also found that selective antagonists of A2B (MRS1706, 10 nM) or A3 (MRS1334, 30 nM) receptors reduced I_GABA run-down, whereas treatment with the specific A1 receptor antagonist DPCPX (10 nM) was ineffective. The selective A2A receptor antagonist SCH58261 (10 nM) reduced or potentiated I_GABA run-down in ≈40% and ≈20% of tested oocytes, respectively. The ADA-resistant, AR agonist 2-chloroadenosine (2-CA) (10 μM) potentiated I_GABA run-down but only in ≈20% of tested oocytes. CGS15943 administration again decreased I_GABA run-down in patch-clamped neurons from either human or rat neocortex slices. I_GABA run-down in pyramidal neurons was equivalent in A1 receptor-deficient and wt neurons but much larger in neurons from A2A receptor-deficient mice, indicating that, in mouse cortex, GABA_A-receptor stability is tonically influenced by A2A but not by A1 receptors. I_GABA run-down from wt mice was not affected by 2-CA, suggesting maximal ARs activity by endogenous adenosine. Our findings strongly suggest that cortical A2–A3 receptors alter the stability of GABA_A receptors, which could offer therapeutic opportunities.