非体外循环冠状动脉旁路移植术后造血干细胞动员的临床研究

目的研究非体外循环冠状动脉旁路移植术(OPCAB)后造血干细胞动员的规律及影响因素。方法对55例择期行OPCAB、近期无急性心肌梗死患者,分别于术前30m in、术后6、12、24、48、72和120h采外周静脉血4m l,CD 34和CD 45双抗标记、流式细胞仪检测造血干细胞计数,同时检测心肌酶谱及肌钙蛋白T的变化。结果冠心病患者术前造血干细胞计数占外周血有核细胞数计数的0.13%±0.12%,OPCAB后明显升高,24h达到峰值(0.34%±0.20%),术后120h回到术前基线水平;吸烟、高血脂、糖尿病对造血干细胞动员无明显影响,但合并高血压病患者外周血造血干细胞动员明显低于无高血压病患者;术后24h造血干细胞动员与肌酸激酶及肌酸激酶同工酶、乳酸脱氢酶、肌钙蛋白T呈正相关(r=0.692,P=0.000;r=0.558,P=0.000;r=0.447,P=0.000;r=0.401,P=0.004)。结论OPCAB术后造血干细胞快速、短暂动员,心肌损伤程度、冠心病危险因素参与了造血干细胞动员的调节。     

Involvement of Ca+ mobilization in the amplifying effect of serotonin on responses of rabbit isolated ear artery to exogenous noradrenaline

Summary The present study examined the involvement of Ca⁺ mobilization in the amplifying effect of serotonin on steady-state responses of rabbit isolated perfused ear artery to exogenous noradrenaline (NA; 0.001–3 ol/l). In contrast to its marked amplifying effect on responses to NA, serotonin in the subconstrictor concentration of 100 nmol/l had no effect on responses to KCl. The Ca⁺-entry blocking drug diltiazem (10 gmol/1) decreased responses to NA; in addition, the amplifying effect of serotonin on responses to NA was reduced by diltiazem. Lowering the concentration of Ca⁺ in the Krebs-Henseleit perfusion solution from 2.5 to 0.25 mmol/l also reduced both responses to NA and the amplifying effect of serotonin. Using the method of Manzini et al. (1982), separate intra- and extracellular Ca⁺-dependent responses to NA were obtained. Serotonin had no effect on intracellular Ca⁺-dependent responses to NA but enhanced extracellular Ca²⁺-dependent NA responses. These results suggest that the amplifying effect of serotonin on responses of rabbit ear artery to exogenous NA involves a selective enhancement of the component of the NA response which is dependent on extracellular Ca⁺; serotonin may increase NA-evoked entry of Ca²⁺ into the vascular smooth muscle cells through receptor-operated Ca⁺ channels. Send offprint requests to A. G. Meehan at the above address     

HD-A+VM_(26)+rhG-CSF动员白血病患儿外周血造血干细胞的初步研究

目的 　评价HD -A +VM2 6+rhG -CSF动员白血病患儿外周血造血干细胞的效果。方法 　 5例高危急性白血病患儿 ,其中急性淋巴细胞白血病 (HR -ALL) 4例 ,急性杂合细胞白血病 (AHL) 1例。年龄 9～ 1 3岁。 5例患儿均采用HD -A +VM2 6+rhG -CSF方案进行动员。在化疗后第 9天应用rhG -CSF ,当白细胞升至 5× 1 0 9/L以上时 ,使用CS - 30 0 0血细胞分离机采集PBSC。结果 　 4例采集 1次 ,1例采集 2次。全部患儿均能耐受该动员方案治疗。MNC值为 5 96 5( 5 0 2～ 7 2 )× 1 0 8/kg。CD3 4+ 细胞量为 4 372 ( 3 95～ 5 6 2 )× 1 0 6/kg。结论 　G -CSF联合大剂量Ara -C和VM2 6的化疗可采集到足够数量的自体外周血干细胞。     

Polymorphism of CD44 Influences the Efficacy of CD34+ Cells Mobilization in Patients with Hematological Malignancies

In the last decade, peripheral blood was the main source of hematopoietic stem cells (HSC) for autologous and allogeneic transplantation. The exact mechanisms of HSC mobilization are still not clear and the efficacy of the procedure is hardly predictable. Ligand-receptor interactions of adhesion molecules, such as SDF1/CXCR4, VLA4/VCAM-1, or CD44/osteopontin, play an important role in homing of HSC in the hematopoietic niche. There is some evidence that disruption of the ligand-receptor complex leads to the egress of HSCs to the peripheral blood. The aim of the present study was the evaluation of constitutive polymorphism of genes encoding cytokines and receptors present in the HSC niche and their impact on the efficacy of mobilization of HSCs in patients with hematological malignancies. We enrolled 110 patients (60 females and 50 males) in the study. The median age of the patients was 55 (range, 22 to 69) years. The group consisted of patients with multiple myeloma (n = 74), non-Hodgkin lymphoma (n = 19), Hodgkin lymphoma (n = 15), or acute myeloid leukemia (n = 2). The mobilization procedures comprised chemotherapy and subsequent granulocyte-colony stimulating factor (G-CSF) at a dose of 10 μg/kg daily. The poor mobilizers group was defined according to Italian National Bone Marrow Transplant Registry criteria: patients with peak CD34⁺ in the peripheral blood < 20/μL or total yield < 2 × 10⁶ CD34⁺ cells/kg body weight in maximum 3 aphereses. Genotyping was performed using standard PCR-based assays. The group of patients (N = 108) who achieved minimal threshold for collections (CD34⁺ at least 10/μL) proceeded to apheresis. The median total yield of CD34⁺ in this group was 5.6 × 10⁶ cells/kg body weight, whereas the median number of cells collected during the first apheresis was 3.3 × 10⁶ cells/kg body weight. Median number of days of G-CSF treatment before first apheresis was 10. Fifteen patients fulfilled the criteria for poor mobilizer. The group of poor mobilizers had higher frequency of TT genotype in rs13347 (CD44) gene (CC+ CT versus TT P = .047). Patients homozygous for T allele had a lower total yield of CD34⁺ cells/kg body weight than the group with allele C (median, 3.7 × 10⁶/kg versus 5.8 × 10⁶/kg; P = .019) and a lower number of CD34⁺ cells gathered during first apheresis (.95 × 10⁶/kg versus 3.3 × 10⁶/kg, P = .04). Multivariate logistic regression analysis revealed that the CD44 TT genotype was the only factor associated with 5-fold higher risk of poor mobilization (P = .037). Polymorphic variants of CXCR4 and VCAM-1 did not significantly influence the efficacy of HSCs mobilization in our group of patients. In conclusion, our results indicate that among investigated single nucleotide polymorphisms (SNPs), only CD44 rs13347 has an impact on the efficacy of HSCs mobilization in patients with hematologic malignancies. CD44 SNPs analysis may be helpful for predicting the poor mobilizers population who may benefit from newer modalities using adhesion molecules inhibitors.     

Diabetes mellitus as a poor mobilizer condition

Hematopoietic stem cell (HSC) transplantation in an effective and curative therapy for numerous hematological malignancies. Mobilization of HSCs from bone marrow (BM) to peripheral blood (PB) followed by apheresis is the gold standard for obtaining HSCs for both autologous and allogeneic stem cell transplantation. After administration of granulocyte-colony stimulating factor (G-CSF), up to 30% of patients fail to mobilize “optimal” numbers of HSCs required for engraftment. This review summarizes the current experimental and clinical evidence that diabetes mellitus is a risk factor for poor mobilization. Diabetes causes a profound remodeling of the HSC niche, resulting in impaired release of HSCs. Experimental studies indicate that hyperglycemia hampers regulation of CXCL12 and clinical studies suggest that diabetes impairs HSC mobilization especially in response to G-CSF, but less to plerixafor. Understanding further the biochemical alterations in the diabetic BM will provide insights into future therapeutic strategies to reverse the so-called “diabetic stem cell mobilopathy”.     

The immediate effect of osteopathic cervical spine mobilization on median nerve mechanosensitivity: A triple-blind,randomized, placebo-controlled trial

Background

Neurodynamics is a clinical medium for testing the mechanical sensitivity of peripheral nerves which innervate the tissues of both the upper and lower limb. Currently, there is paucity in the literature of neurodynamic testing in osteopathic research, and where there is research, these are often methodologically flawed, without the appropriate comparators, blinding and reliability testing.

内皮祖细胞的动员及其在治疗缺血性心肌病中的应用

内皮祖细胞是一类能分化成血管内皮细胞的前体细胞,在成人血管发生中起着重要作用。从骨髓动员内皮祖细胞,增加循环内皮祖细胞的数量,对治疗缺血性心肌病有重要的临床意义。     

Engraftment after autologous hematopoietic stem cell transplantation in patients mobilized with Plerixafor: A retrospective,multicenter study of a large series of patients

Plerixafor (PLX) appears to effectively enhance hematopoietic stem-cell mobilization prior to autologous hematopoietic stem cell transplantation (auto-HCT). However, the quality of engraftment following auto-HCT has been little explored. Here, engraftment following auto-HCT was assessed in patients mobilized with PLX through a retrospective, multicenter study of 285 consecutive patients. Information on early and 100-day post-transplant engraftment was gathered from the 245 patients that underwent auto-HCT. The median number of PLX days to reach the stem cell collection goal (≥2 × 10⁶ CD34⁺ cells/kg) was 1 (range 1–4) and the median PLX administration time before apheresis was 11 h (range 1–18). The median number of apheresis sessions to achieve the collection goal was 2 (range 1–5) and the mean number of CD34⁺ cells collected was 2.95 × 10⁶/kg (range 0–30.5). PLX administration was safe, with only 2 mild and transient gastrointestinal adverse events reported. The median time to achieve an absolute neutrophil count (ANC) >500/μL was 11 days (range 3–31) and the median time to platelet recovery >20 × 10³/μL was 13 days (range 5–69). At 100 days after auto-HCT, the platelet count was 137 × 10⁹/L (range 7–340), the ANC was 2.3 × 10⁹/L (range 0.1–13.0), and the hemoglobin concentration was 123 g/L (range 79–165). PLX use allowed auto-HCT to be performed in a high percentage of poorly mobilized patients, resulting in optimal medium-term engraftment in the majority of patients in whom mobilization failed, in this case mainly due to suboptimal peripheral blood CD34⁺ cell concentration on day +4 or low CD34⁺ cell yield on apheresis.     

An innovation in stem cell harvesting: Heparin use

Background and ObjectivesStem cell transplantation is a growing treatment strategy for most malignant and non- malignant hematological diseases. Plerixafor and granulocyte colony stimulating factor (G-CSF) are usually used in mobilization regimens to increase the CD34⁺ cell count in the harvest. Heparin is a sulphated glycosaminoglycated polymer with 12−15 kDa mass. Heparin inhibits the CXCR4/SDF1 axis, as does plerixafor. In this study, our aim was to investigate the effect of using heparin on stem cell mobilization and harvesting.Materials and MethodsWe administered 5000 units of unfractioned heparin intravenously in 150 mL (mL) of isotonic sodium chloride solution, 15 min before the stem cell harvesting procedure to 141 patients who underwent bone marrow transplantation between the years of 2018 and 2019 at our Stem Cell Transplantation Unit. Thirty patients were included as a control group, and they were not given heparin. The study population included patients with multiple myeloma and lymphoma equally in each group.ResultsIn all patients hematopoeitic stem cells were successfully harvested in a single cycle of apheresis. In multiple myeloma patients who received heparin, the mean collected CD34⁺ cell number was 8 × 10⁶/kg, and the mean CD34⁺ cell number yield was 12,555/μl. In the control group, the mean collected CD34⁺ cell number was 4,2 × 10⁶/kg, and mean CD34⁺ cell number in yield was 492/μl. In lymphoma patients who received heparin, the mean collected CD34⁺ cell number was 6,8 × 10⁶/kg, and the mean CD34⁺ cell number was 1421/μl. In the control group the mean collected CD34⁺ cell number was 4,3 × 10⁶/kg, and the mean CD34⁺ cell number was 358/μl. The effect of heparin on the collected stem cell number in both myeloma and lymphoma patients was statistically significant (p < 0.01).ConclusionsOur results have shown that heparin increases harvested stem cell numbers significantly. Heparin may be a promising agent for stem cell harvesting.