自体外周血和骨髓干细胞移植对缺血性下肢血管疾病患者缺血状态改善的临床研究

目的观察自体外周血干细胞(PBSC)和骨髓干细胞(BMSC)移植在下肢缺血性疾病患者中血管新生和改善缺血状态方面的疗效。方法2004年9月至2006年10月对解放军第463医院细胞治疗中心的170例自愿接受自体干细胞移植的下肢缺血性血管疾病患者随机分为PBSC移植组(94例)和BMSC移植组(76例)。第1,4,12,24周对移植前后临床症状主观指标和仪器检测客观指标结果进行综合评估,比较两组患者血管新生及下肢缺血改善情况。结果PBSC组单个核细胞(MNC)、CD34 、CD133 细胞总数明显多于BMSC组(P<0.01);干细胞移植前后疼痛、冷感、行走间距、溃疡、坏疽等临床症状对照研究表明,PBSC组缺血症状改善较BM-SC组出现早,症状缓解程度较BMSC组更明显,但两组间差异无显著性意义(P(0.05);两组患者移植前后的踝肱指数(ABI)、经皮氧分压、足底深感觉、皮温仪结果比较差异无显著性意义。结论自体PBSC和BMSC移植技术在下肢缺血性疾病治疗中均可建立新生血管,有效改善下肢缺血状态,治疗缺血性下肢血管疾病各有其不可替代的优势,可根据患者具体情况选择干细胞来源。     

自体外周血干细胞移植治疗糖尿病足26例临床研究

目的　观察自体外周血干细胞移植治疗糖尿病足的临床疗效及相关影响因素。方法　自 2 0 0 3- 112 0 0 4 - 0 7解放军第 4 6 3医院对 2 6例自愿接受自体外周血干细胞移植的糖尿病足患者进行外周血干细胞动员 ,给予rhG -CSF每天 4 5 0 6 0 0 μg ,皮下注射 ,第 6天用COBE 6 1血细胞分离机单采干细胞 ,总量 82 14 8mL。将干细胞悬液按 3cm×3cm距离进行缺血肢体移植术 ,移植后观察各项指标综合评估。结果　移植后 314d 2 3例患者疼痛明显缓解 ,2 7d2 4例患肢冷感有不同程度的改善 ,4 12周 4例溃疡明显好转 ,踝压指数 (ABI) 6例患者改善。 2例DSA下肢动脉造影结果显示新侧支血管形成明显增加。未出现并发症和不良反应。结论　自体外周血干细胞移植治疗糖尿病足是一种简便、安全、有效的治疗方法     

干细胞移植治疗下肢缺血性疾病的研究进展

下肢缺血性疾病严重危害人类的健康,药物治疗疗效甚微,且有些病人无法行血运重建治疗。干细胞移植促进缺血下肢的血管新生,对缺血性疾病的治疗已日益受到关注。本文就干细胞移植治疗下肢缺血性疾病的应用作一综述。     

自体外周血干细胞移植治疗下肢缺血性疾病术前行干细胞动员引发心脑血管危险因素的探讨

目前。自体外周血干细胞移植已经成为治疗下肢缺血性疾病的一种有效手段。但移植之前的干细胞动员还是存在一定风险的,尤其是潜伏在心脑血管系统中的危险因素最为引人关注。现就干细胞移植动员期间发生了心脑血管并发症的病例进行分析。以便更好的提高该项技术的安全性。     

内皮祖细胞和单个核细胞移植对糖尿病大鼠血管病变疗效的比较

目的比较内皮祖细胞（EPC）和单个核细胞（MNC）移植改善糖尿病大鼠后肢血管病变的疗效。方法将大鼠EPC和外周血MNC标记后分别植入糖尿病大鼠缺血后肢。对照组仅植入PBS溶液。ELISA法检测VEGF的含量，免疫组化法计数毛细血管数目。结果EPC和MNC均能在缺血后肢分化为毛细血管。移植后EPC组和MNC组大鼠缺血肢体局部VEGF含量高于对照组，毛细血管数目也高于对照组。但EPC组和MNC组VEGF含量和毛细血管数目无明显差异。结论移植EPC及含有相当数量EPC的MNC对糖尿病大鼠后肢血管病变的改善作用相似。EPC可能在外周血单个核细胞移植中发挥主要作用。     

造血干细胞移植治疗下肢血管病变的研究现状

近年来，随着干细胞技术研究的快速发展，造血干细胞移植用于下肢缺血性疾病的治疗，促进缺血肢体的血管新生和侧肢循环形成，使缺血的肢体得到改善的新治疗方法已成为热点。     

造血干细胞移植治疗下肢血管病变的研究现状

近年来,随着干细胞技术研究的快速发展,造血干细胞移植用于下肢缺血性疾病的治疗,促进缺血肢体的血管新生和侧肢循环形成,使缺血的肢体得到改善的新治疗方法已成为热点[1].     

骨髓干细胞动员促进兔缺血后肢血管新生的实验研究

目的观察干细胞动员剂动员骨髓干细胞后,促进兔缺血后肢血管新生的情况。方法切断股动脉及其分支,制作兔后肢缺血模型;实验组兔皮下注射重组人粒细胞集落刺激因子(rhG-CSF)动员兔自体干细胞释放和迁移至缺血组织;对照组给予生理盐水。术后4周,行腹主动脉造影观察侧支循环形成情况,并取内收肌和腓肠肌行病理切片HE染色,应用图像分析系统统计血管密度。结果实验组兔侧支循环血管数量和血管密度大于对照组(P<0.05),缺血状态得到改善。结论骨髓干细胞动员可促进血管新生,增加缺血下肢血液灌注,改善肢体缺血状态。     

自体外周血干细胞移植治疗糖尿病下肢血管病变与糖尿病足的疗效观察

目的观察自体外周血干细胞移植治疗糖尿病下肢血管病变与糖尿病足的临床疗效。方法应用自体外周血干细胞移植治疗8例糖尿病周围血管病与糖尿病足患者,男5例,女3例。全部患者均给予“干细胞动员”5天。第5天采集外周血干细胞,将单个核细胞悬液按3cm×3cm间距进行缺血肢体移植术。术后第1~6个月定期观察各项指标并进行综合评估。结果所有患者疼痛、肢体凉冷感均有不同程度的缓解,行走距离延长,踝肱指数(ABI)有不同程度的改善,3例患者复查有新生血管形成。结论自体外周血干细胞移植治疗糖尿病周围血管病与糖尿病足是一种有效、安全的新方法,能改善患者的生活质量并避免截肢的痛苦。     

儿童难治性自身免疫性疾病自体外周血干细胞动员采集和分选的临床研究

目的 探讨儿童难治性自身免疫性疾病进行自体外周血干细胞动员采集的安全性和CD34+细胞分选纯化的可行性及其临床意义.方法 8例儿童难治性自身免疫性疾病,包括4例系统性红斑狼疮、2例皮肌炎,1例幼年型类风湿关节炎和1例多发性硬化,予行CD34+细胞纯化的自体外周血干细胞移植.首先采用环磷酰胺(CTX)联合粒细胞集落刺激因子(G-CSF)方案动员外周血干细胞,然后采用CS-3000血细胞分离机采集外周血,通过CliniMACS细胞分选仪分选自体外周血CD34+细胞,将其用保养液配置冻存于-80℃冰箱.采用非清髓内去除T的预处理方案,即卡氮芥+足叶乙苷+阿糖胞苷+马法兰+抗胸腺球蛋白(ATG)或CTX+ATG或CTX+马法兰+ATG,于第0天回输自体外周血CD34+细胞.结果 儿童能够耐受自体外周血干细胞动员采集过程,无动员相关死亡,动员后获得的单个核细胞数和CD34+细胞数的平均值分别为8.35×108/kg和7.92×106/kg,纯化后的白体外周血CD34+和CD3+细胞数的平均值分别为6.28×106/kg和0.71×105/kg.回输后中性粒细胞和血小板的植入中位时间分别为+11d和+15 d.结论 经CTX联合G-CSF方案可动员出足量的外周血干细胞,经CS-3000血细胞分离机采集可获得足够的单个核细胞,在动员过程中原发病无明显进展恶化,患儿能耐受动员方案,采集过程顺利安全;经CliniMACS细胞分选仪分选的自体外周血CD34+细胞纯度高,移植后造血恢复;采用CD34+细胞纯化的自体外周血移植治疗是常规治疗无效的儿童难治性自身免疫性疾病的可选择治疗措施之一.     

内皮祖细胞移植对糖尿病兔下肢缺血的病理及血管内皮生长因子的影响

目的探讨内皮祖细胞(EPC)移植治疗糖尿病兔下肢缺血的病理及缺血肌肉中血管内皮生长因子(VEGF)的变化。方法2006年1月至9月在哈尔滨医科大学附属第二医院内分泌科将30只健康日本大耳白兔随机分为3组,即糖尿病磷酸盐缓冲液(PBS)对照组(A组)8只、糖尿病内皮祖细胞移植治疗组(B组)14只、正常血糖内皮祖细胞移植治疗组(C组)8只。骨髓来源的内皮祖细胞经体外扩增培养7d后,通过肌内注射进行细胞移植,用病理改变及肌浆中VEGF的变化评价治疗效果。结果EPC移植后14d,病理示3组毛细血管密度分别为(9.29±1.63)个/视野、(12.60±2.16)个/视野、(12.51±1.56)个/视野;血管数/肌束数分别为0.66±0.05,0.83±0.11,0.90±0.13;肌浆中VEGF质量分数[每克肌肉中VEGF因子的质量(ng)]分别为0.22±0.05,0.30±0.07,0.31±0.08;糖尿病内皮祖细胞移植治疗组与糖尿病PBS对照组比较,毛细血管密度、血管数/肌束数、VEGF值差异均有显著性意义,P<0.05。结论EPC移植能有效治疗糖尿病兔下肢缺血。     

内皮祖细胞移植治疗糖尿病兔下肢缺血的影像学改变

目的探讨内皮祖细胞（EPC）移植治疗糖尿病兔模型下肢缺血的疗效。方法将日本大耳白兔随机分3组：糖尿病PBS对照组（A组）、糖尿病内皮祖细胞移植治疗组（B组）、正常血糖内皮祖细胞移植治疗组（C组）。骨髓来源的内皮祖细胞经体外扩增培养7d后,通过肌内注射进行细胞移植。结果EPC移植后14d,彩超示B组兔缺血侧／正常侧下肢胫前动脉收缩期峰值流速比值明显增加,动脉造影示该组缺血侧下肢动脉显影血管数多于对照组（P〈0．05）。结论EPC移植能有效治疗糖尿病兔模型下肢缺血。     

自体外周血干细胞移植治疗62例缺血性下肢血管病的临床研究

目的　观察自体外周血干细胞(PBSC)移植治疗缺血性下肢血管病的临床疗效及相关影响因素。方法　62例自愿接受自体PBSC移植的患者中糖尿病足34例,动脉硬化闭塞症22例,血栓闭塞性脉管炎4例,结节性多动脉炎1例,雷诺病1例。发生足部溃疡或坏疽38例,合并感染14例。累及双下肢病变52例,其中3例患者移植前已行一侧肢体截肢术;单肢病变10例,共移植111条下肢。全部患者均给予重组人粒细胞集落刺激因子450~600μg/d,皮下注射,行PBSC动员5d,第6天用血细胞分离机采集单个核细胞(MNC),总量82~148ml,MNC数量(7182~2246)×109/L。将MNC悬液按3cm×3cm距离进行缺血肢体移植术,术后第1天至6个月定期观察各项指标综合评估。结果　PBSC移植后7~30d54例(871%)疼痛有不同程度的缓解,7~30d56例(903%)患肢冷感明显改善,4~16周16例(400%)溃疡或足趾坏疽好转,感染得到控制;踝肱指数12例(343%)有不同程度的改善,26例(423%)经皮氧分压改善,5例数字减影下肢动脉造影结果显示新侧支血管形成明显增加。2例糖尿病足合并脑梗死患者在干细胞动员期间症状加重,经治疗好转,其余患者未出现并发症和明显不良反应。结论　自体PBSC治疗缺血性下肢血管病是一种安全、有效的手段,尤其对于下肢动脉没有流出道的患者有特殊意义,可使一部分患者免除截     

Adhesion and incorporation of lacZ-transduced endothelial cells into the intact capillary wall in the rat.

Use of the capillary bed of skeletal muscle as an in vivo recipient site to transplant autologous endothelial cells that have undergone gene transfer ex vivo has considerable potential as a technique of somatic gene therapy. Here we document a previously unrecognized capacity of endothelial cells to adhere and incorporate spontaneously into confluent endothelial cell monolayers in vitro and in vivo. This spontaneous adhesion and incorporation of endothelial cells enabled us to seed lacZ-transduced endothelial cells into the wall of skeletal muscle capillaries of the hindlimb of the rat. Certain transduced endothelial cells became incorporated within the capillary wall, whereas others remained within the capillary lumen where they formed focal, electron-dense, contacts with host endothelium. lacZ expression in the capillary bed was documented for up to 1 month after transplantation. Use of the intact capillary bed of skeletal muscle as an in vivo recipient site for transduced, autologous endothelial cells holds promise as a strategy for somatic gene therapy to treat various genetic and acquired human diseases.     

Consolidation therapy with autologous blood stem cell transplantation in a patient with primary plasma cell leukaemia

Primary plasma cell leukaemia (PPCL) is a rare form of plasma cell dyscrasia. Conventional melphalan-based treatment is often ineffective, with a reported median survival of 2-7 months only. We report a 53-year-old man with PPCL who was treated with four cycles of combination chemotherapy including vincristine, adriamycin and dexamethasone that resulted in a good partial remission. High-dose melphalan 200 mg/m2 and autologous peripheral blood stem cell (PBSC) rescue was then given 6 months after diagnosis. Maintenance interferon-alpha was started 8 weeks after transplantation with good drug compliance. Complete remission was achieved and molecular remission was documented 11 months after autologous PBSC transplantation. In conclusion, high-dose therapy followed by autologous stem cell rescue is a feasible option for PPCL that can result in a reasonably sustained remission.     

Lack of additional benefit of intracoronary transplantation of autologous peripheral blood stem cell in patients with acute myocardial infarction.

BACKGROUND: Recently the potential of myocardial repair by transplantation of autologous bone marrow stem cells has been suggested. Whether the additional intracoronary transplantation of autologous peripheral blood stem cells (PBSC), which were mobilized by granulocyte-colony-stimulating factor (G-CSF), could safely improve myocardial function in patients with acute myocardial infarction (AMI) was investigated. METHODS AND RESULTS: Seventy-three patients with AMI who had successfully undergone percutaneous coronary intervention (PCI) were enrolled in the present prospective nonrandomized open-labeled study. Ten patients with elective PCI received G-CSF for 4 days followed by intracoronary PBSC transplantation. Thirty-two patients with primary PCI and 31 patients with recent AMI and elective PCI served as controls. The left ventricular (LV) function was evaluated using echocardiography and magnetic resonance imaging. G-CSF and intracoronary transplantation of PBSC did not incur any periprocedural myocardial damage. After 6 months, the LV ejection fraction was significantly improved in the cell therapy group. For 2 years of the follow-up period, there was no adverse clinical events, except one asymptomatic in-stent restenosis. However, comparable improvement of the LV ejection fraction was also identified in the primary PCI and elective PCI control groups. CONCLUSIONS: In the present study, additional intracoronary infusion of PBSC was safe and feasible for the patients with AMI who had undergone PCI, but did not lead to a significant improvement in LV function compared to standard reperfusion treatment.     

Long-term survival in primary plasma cell leukemia after therapy with VAD, autologous blood stem cell transplantation and interferon-alpha.

Primary plasma cell leukemia (PCL) is a rare form of plasma cell neoplasm with a poor prognosis. Conventional melphalan-based treatments have been most disappointing. We report the case of a 62-year-old man with a primary form of PCL treated with VAD combination achieving an objective response, and who received high-dose melphalan and autologous peripheral blood stem cell (PBSC) transplantation followed by interferon-alpha. During the remission time, lasting for 3 years, an infiltration by large granular lymphocytes (LGL) was noted in peripheral blood. However, when the number of LGL declined, a bone marrow relapse was observed. The treatment for PCL and the possible role of these LGL on tumor cell control after autologous PBSC transplantation are discussed.     

Addition of peripheral blood stem cells collected without mobilization techniques to transplanted autologous bone marrow did not hasten marrow recovery following myeloablative therapy.

A randomized prospective trial was conducted to determine if the addition of cryopreserved autologous peripheral blood stem cells (PBSC) collected without mobilization techniques to autologous cryopreserved bone marrow for patients receiving an autologous bone marrow transplant (ABMT) affected the time to marrow function recovery. Thirty-five evaluable patients with various malignancies were studied. Sixteen received PBSC + ABMT and 19 received ABMT alone. The PBSC were collected with 4 h leukapheresis procedures on 3 consecutive days. No manipulations to increase the number of circulating stem cells were used during the collections. The median time to recover 0.5 x 10(9)/l circulating granulocytes was 20 days after transplantation in the ABMT group and 27 days in the PBSC + ABMT group (p = 0.12). The median time to recover 20 x 10(9)/l platelets was 22 days after transplantation in the ABMT group and more than 27 days in the PBSC + ABMT group (p = 0.29). The day of discharge from the hospital was earlier for the ABMT group (median 29 days) than the PBSC + ABMT group (median 35 days, p = 0.03). We did not find that the addition of non-mobilized PBSC to infused autologous marrow accelerates marrow recovery.     

Roles of endogenous monocyte chemoattractant protein-1 in ischemia-induced neovascularization

OBJECTIVES: We sought to investigate the role of endogenous monocyte chemoattractant protein (MCP)-1 in ischemia-induced neovascularization. BACKGROUND: Roles of inflammatory changes including macrophage infiltration are suggested in ischemic neovascularization. METHODS: Unilateral hindlimb ischemia was induced by excising surgically the entire femoral artery and vein in mice. Immediately after operation, plasmid deoxyribonucleic acid encoding a dominant negative mutant of MCP-1 (7ND) or the empty plasmid (mock) was injected into the ipsilateral thigh adductor muscle. RESULTS: In mock-treated mice, MCP-1 was upregulated transiently in ischemic hindlimb peaking at day 3. Serial laser Doppler blood flow (LDBF) analysis showed an abrupt decrease in blood flow, followed by a recovery to the near-normal levels in mock-treated mice; 7ND treatment had no effects on the initial decrease in LDBF but deteriorated the recovery. At day 3, macrophage infiltration and inductions of tumor necrosis factor (TNF)-alpha and vascular endothelial growth factor (VEGF) were prominent in the ischemic adductor muscle in mock-treated mice; 7ND treatment significantly reduced macrophage infiltration and suppressed TNF-alpha and VEGF inductions in response to ischemia. At day 21, postmortem angiography and anti-CD31 immunohistostaining revealed well-developed collateral vessels and capillary formation, respectively, in the ischemic muscle of mock-treated mice; 7ND overexpression remarkably suppressed the collateral vessel formation and capillary formation. CONCLUSIONS: Endogenous MCP-1 may play a role in ischemia-induced neovascularization by recruiting macrophages that activate TNF-alpha and VEGF inductions.