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1.
目的 探讨环孢菌素A(CsA)治疗对再生障碍性贫血(再障)患者骨髓单个核细胞Fas(CD95)抗原表达及凋亡的影响作用。方法 用流式细胞术检测骨髓(BM)CD34^ 细胞、CD34^-细胞Fas抗原的表达,用原位末端标记法(TUNEL)测定骨髓单个核细胞凋亡率。结果 应用CsA治疗再障2个月时的总有效率为40%。再障患者骨髓CD34^ 细胞百分率显著低于正常对照(P<0.05),再障患者骨髓CD34^ 细胞Fas表达率显著高于正常对照(P<0.01),用CsA治疗后,再障患者骨髓单个核细胞凋亡率显著低于治疗前(P<0.01)。CsA治疗前CD34^ 细胞Fas表达率与骨髓单个核细胞凋亡率呈正相关。结论 CsA治疗可降低再障患者骨髓单个核细胞凋亡率,CD34^ 细胞Fas的异常表达与再障的发病有关。  相似文献   

2.
背景:骨髓间充质干细胞具有免疫调节作用,可以影响淋巴细胞和其他免疫细胞的功能,同时其分泌的因子尚有支持造血的作用。而再生障碍性贫血的发生、发展与T细胞功能亢进引起的造血干、祖细胞凋亡密切相关。目的:比较骨髓间充质干细胞和全骨髓细胞治疗免疫介导再生障碍性贫血小鼠的疗效。方法:建立BALB/C小鼠免疫介导再生障碍性贫血模型。将BALB/C小鼠随机分成正常组、对照组、注射骨髓间充质干细胞和注射全骨髓细胞组。观察骨髓间充质干细胞和全骨髓细胞对实验动物的疗效及骨髓的造血组织容量、脂肪细胞和血窦结构。结果与结论:再生障碍性贫血模型小鼠至第20天,对照组全部死亡,骨髓间充质干细胞组4例死亡,全骨髓细胞组1例死亡。②骨髓间充质干细胞组和全骨髓细胞组存活小鼠,其体质量变化趋势、血细胞计数、骨髓造血功能恢复均无显著性差异。③检测骨髓间充质干细胞组和全骨髓细胞组脾脏CD4+T细胞、CD8+T细胞、CD4+/CD8+的比值示无差异,与正常小鼠相比,骨髓间充质干细胞组外周血CD4+T细胞、CD8+T细胞水平有下降趋势。全骨髓细胞组CD8+T细胞水平明显下降。提示,单独使用骨髓间充质干细胞治疗能够使部分再生障碍性贫血模型小鼠症状改善、造血恢复。骨髓间充质干细胞治疗有效率不如全骨髓细胞,但两组无显著性差异。  相似文献   

3.
骨髓间充质干细胞治疗再生障碍性贫血模型鼠的机制   总被引:1,自引:1,他引:0  
背景:骨髓间充质干细胞在体外抑制再生障碍性贫血T淋巴细胞的活化与增殖,并有支持骨髓造血作用.目的:观察骨髓间充质干细胞对再生障碍性贫血模型鼠存活率的影响,探讨其治疗再生障碍性贫血的机制.设计、时间及地点:随机对照动物实验,于2009-01/04在安徽医科大学第一附属医院中心实验室完成.材料:清洁级雌性BALB/C小鼠40只,8~10周龄,体质量16~18 g,作为受体.清洁级DBA/2小鼠5只,6~10周龄,体质量15~19 g为淋巴细胞供体.方法:取BALB/c小鼠骨髓进行间充质干细胞体外增殖培养,传至3~5代的细胞用于实验.建立免疫介导再生障碍性贫血小鼠模型.40只BALB/C小鼠随机分为3组,治疗组(n=15):造模后将1×106个骨髓间充质干细胞输注到治疗组小鼠体内进行治疗;模型组(n=15):仅造模不进行任何治疗;对照组(n=10):不进行任何干预.主要观察指标:分别于第1,2,3,4周观察各组小鼠的存活率;流式细胞仪及酶联免疫吸附法测定骨髓间充质干细胞治疗后第28天各组小鼠T淋巴细胞亚群及血浆肿瘤坏死因子α的表达.结果:①模型组小鼠3只于第2周内死亡,7只于第3周内死亡,2只于第4周内死亡,28天存活率20%;治疗组小鼠4只于第3周内死亡,28天存活率73%.②与模型组相比,治疗组和对照组CD4的表达增高(27.12±4.23)%,(32.18±6.73)%,(38.44±7.88)%,P<0.01];CD8和肿瘤坏死因子α表达均降低[CD8:(34.42±5.13)%,(27.33±5.09)%,(23.31±5.24)%,P<0.01:肿瘤坏死因子α:(47.27±10.1 1),(12.34±5.87),(9.31±3.77)ng/L,P<0.011.结论:骨髓间充质干细胞可以提高再生障碍性贫血小鼠存活率,其治疗机制可能与调节T淋巴细胞亚群及抑制T淋巴细胞分泌的肿瘤坏死因子α有关.  相似文献   

4.
目的 探讨再生障碍性贫血(AA)的发病机制.方法 采用流式细胞术对25例AA患者进行骨髓CD34+细胞及外周血T淋巴细胞亚群进行检测,10例健康者骨髓以及25例健康者外周血T淋巴细胞亚群做为对照.结果 AA患者与健康对照组比较,骨髓CD34+细胞表达减低,差异有统计学意义(P<0.05);CD3+细胞略增高,差异有统计学意义(P<0.05),CD4+细胞减低,差异有统计学意义( P<0.05),CD8+细胞明显增高,差异有统计学意义(P<0.01),CD4+/CD8+比值明显减低,差异有统计学意义(P<0.01);25例AA患者中,13例AA患者存在细胞免疫异常(CD4+/CD8+比值倒置);17例患者骨髓CD34+细胞数量减少;6例患者二者均表达异常;1例早期AA患者二者表达均正常.结论 大部分AA患者骨髓造血干祖细胞数量减少;部分AA患者细胞免疫功能异常;也有部分患者二者均异常.  相似文献   

5.
目的 探讨细胞粘附分子(CAMs)CD11a、CD49d在再生障碍性贫血(AA)患者的表达特点及临床应用.方法 采用碱性磷酸酶抗碱性磷酸酶法(APAAP法)测定26例慢性再生障碍性贫血(CAA)、6例急性再牛障碍性贫血(AAA)患者和20例非血液病的外科手术患者骨髓及外周血单个核细胞(MNC)粘附分子CD11a、CD49d表达的阳性细胞百分率.结果慢性再生障碍性贫血患者骨髓及外周血MNC的CD11a及骨髓MNC的CD49d表达的阳性细胞百分率较对照组降低,外周血MNC的CD49d表达与对照组无显著差异.急性再生障碍性贫血患者骨髓及外周血MNC的CD11a及骨髓、外周血MNC的CD49d表达与对照组无显著差异.结论 细胞粘附分子表达在急、慢性再生障碍性贫血有所不同.细胞粘附分子表达降低在慢性再生障碍性贫血的发病中发挥了一定作用,纠正再障患者粘附分子的异常表达,可以改善其骨髓造血动能.  相似文献   

6.
为了观察骨髓增生异常综合征 (MDS)患者骨髓CD34+ 细胞Fas ,FasL和Bcl 2的表达和凋亡情况并探讨这些抗原的表达和细胞凋亡的关系。采用流式细胞术测定了 2 6例MDS和 10例急性髓系白血病 (AML)患者及 6例非血液病患者 (对照 )骨髓CD34+ 细胞的Fas,FasL和Bcl 2表达率和细胞凋亡率。结果显示 ,各型MDS患者CD34+ 细胞Fas和FasL表达率较对照组明显增加 (P <0 .0 1) ,Bcl 2的表达率除难治性贫血 /环形铁粒幼细胞性难治性贫血 (RA/RAS)与对照组无显著差异 (P >0 .0 5 )外 ,难治性贫血伴有原始细胞增多 (RAEB)和转变中的难治性贫血伴原始细胞增多 (RAEB t)患者均明显高于对照组 (P <0 .0 1) ;各型MDS间CD34+ 细胞Fas的表达率相近 ,而Bcl 2的表达率却存在非常显著性差异 ,即RA/RAS 相似文献   

7.
背景:骨髓间充质干细胞具有免疫调节作用,可以影响淋巴细胞和其他免疫细胞的功能,同时其分泌的因子尚有支持造血的作用。而再生障碍性贫血的发生、发展与T细胞功能亢进引起的造血干、祖细胞凋亡密切相关。目的:比较骨髓间充质干细胞和全骨髓细胞治疗免疫介导再生障碍性贫血小鼠的疗效。方法:建立BALB/C小鼠免疫介导再生障碍性贫血模型。将BALB/C小鼠随机分成正常组、对照组、注射骨髓间充质干细胞和注射全骨髓细胞组。观察骨髓间充质干细胞和全骨髓细胞对实验动物的疗效及骨髓的造血组织容量、脂肪细胞和血窦结构。结果与结论:再生障碍性贫血模型小鼠至第20天,对照组全部死亡,骨髓间充质干细胞组4例死亡,全骨髓细胞组1例死亡。②骨髓间充质干细胞组和全骨髓细胞组存活小鼠,其体质量变化趋势、血细胞计数、骨髓造血功能恢复均无显著性差异。③检测骨髓间充质干细胞组和全骨髓细胞组脾脏CD4+T细胞、CD8+T细胞、CD4+/CD8+的比值示无差异,与正常小鼠相比,骨髓间充质干细胞组外周血CD4+T细胞、CD8+T细胞水平有下降趋势。全骨髓细胞组CD8+T细胞水平明显下降。提示,单独使用骨髓间充质干细胞治疗能够使部分再生障碍性贫血模型小鼠症状改善、造血恢复。骨髓间充质干细胞治疗有效率不如全骨髓细胞,但两组无显著性差异。  相似文献   

8.
目的:检测川芎嗪对急性放射性损伤小鼠骨髓单个核细胞淋巴细胞功能相关抗原-1(LFA-1)表达变化的影响,探讨放射损伤后骨髓造血功能恢复的可能机制.方法:42只小鼠随机分为正常组、对照组和川芎嗪组.对照组和川芎嗪组首先进行总剂量为6.0 Gy 60Coγ射线一次性全身均匀照射;照射后川芎嗪组每只小鼠立即喂饲盐酸川芎嗪注射液2 mg,对照组喂饲生理盐水0.2 ml,均每日2次.照射后第7、14和21 d测定各组小鼠的外周血细胞计数、骨髓单个核细胞计数、骨髓巨核细胞计数、骨髓造血组织面积和骨髓单个核细胞LFA-1的表达水平.结果:照射后第7、14和21 d川芎嗪组外周血细胞计数、骨髓单个核细胞计数、巨核细胞计数和骨髓造血组织面积均显著高于对照组(P<0.05或 P<0.01);7 d和14 d川芎嗪组及对照组骨髓单个核细胞LFA-1表达水平呈进行性增高,且川芎嗪组LFA1表达水平显著高于对照组(P<0.05或P<0.01);21 d川芎嗪组骨髓单个核细胞LFA-1的表达水平已开始下降,且低于对照组(P<0.05).结论:急性放射损伤早期骨髓单个核细胞LFA-1呈高表达;川芎嗪对放射损伤小鼠骨髓造血功能恢复起促进作用的机制之一可能是通过增强受照射小鼠骨髓单个核细胞LFA-1表达实现.  相似文献   

9.
背景:骨髓间充质干细胞具有成脂和成骨分化潜能,再生障碍性贫血患者骨髓间充质干细胞可能存在成脂和成骨分化异常。目的:观察再生障碍性贫血患者骨髓间充质干细胞增殖及分化潜能与健康人的差异。方法:选择哈尔滨市第一医院血液病肿瘤研究所收治的再生障碍性贫血患者5例,另选5例健康成人作为对照组;采用全骨髓培养法分离骨髓间充质干细胞。结果与结论:采用全骨髓培养法成功分离出骨髓间充质干细胞;与对照组相比,再生障碍性贫血组骨髓间充质干细胞的细胞形态和免疫表型无显著差异,皆表达CD73、CD90、CD105,不表达CD34、CD45和HLA-DR;再生障碍性贫血组骨髓间充质干细胞增殖能力显著低于对照组(P<0.05),成脂肪细胞分化诱导率高于对照组(P<0.05),PPARγmRNA和FABP4 mRNA表达量均高于对照组(P<0.05);而成骨细胞分化诱导率低于正常对照组(P<0.05),ALPmRNA和BGLAP mRNA表达量分别低于对照组(P<0.05)。提示再生障碍性贫血组骨髓间充质干细胞分化潜能存在成脂肪细胞和成骨细胞分化的失衡。  相似文献   

10.
目的:探讨再生障碍性贫血患者治疗前后骨髓和外周血标本单个核细胞中幼红巨噬细胞黏附蛋白(EMP)基因的表达。方法:抽取18例再生障碍性贫血患者治疗前后的骨髓和外周血标本,另选10例健康骨髓捐献者为正常对照组,均分离骨髓和外周血单个核细胞,分别提取细胞总RNA,用RT-PCR方法检测EMP mRNA。结果:再生障碍性贫血患者的骨髓和外周血中EMP表达水平分别显著低于正常对照组(P<0.05),而治疗后的表达显著增高(P<0.05)。结论:EMP mRNA可为再生障碍性贫血的诊断与治疗提供依据。  相似文献   

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目的 探讨阵发性睡眠性血红蛋白尿症(PNH)患CD34^ CD59^ 细胞的特性及PNH克隆呈优势造血的可能原因,以探索PNH发病的内在机制。方法 用免疫磁珠吸附法富集纯化CD34^ 细胞,再用流式细胞仪分选出PNH患的CD34^ CD59^ 细胞、CD34^ CD59^ 细胞及正常对照CD34^ 细胞。分别进行体外扩增液体培养2周,并对扩增前、后的细胞进行半固体培养。结果 ①PNH患CD34^ CD59^ 细胞与正常对照CD34^ 细胞形成集落形成单位(CFU)均在第7天达到扩增高峰,并且扩增后的细胞仍能保持CD59抗原,无GPI锚连蛋白的丢失。②正常对照的CD34^ 细胞在生存、增殖、形成CFU及扩增能力上均明显强于FHN患的CD34^ CD59^ 细胞及CD34^ CD59^-细胞.③PNH患CD34^ CD59^ 细胞及CD34^ CD59^ 细胞体外半固体培养,其形成CFU的能力无明显差异。④PNH患CD34^ CD59^ 细胞及CD34^ CD59^ 细胞在SCF IL3 IL6 FL Tpo及SCF IL3 IL6 FL Tpo Epo组合下液体培养,其生存、增殖、扩增等能力上均无明显差异。但在SCF IL3 IL6 FL Tpo Epo GM-CSF组合下液体培养,CD34^ CD59^ 细胞的生存、增殖、扩增能力均明显强于CD34^ CD59^ 细胞。结论 ①正常对照的CD34^ 细胞在生存、增殖、形成CFU及扩增能力上均明显强于PNH患的CD34^ CD59^ 细胞。②PNH患CD34^ CD59^ 细胞及CD34^ CD59^ 细胞体外半固体培养,以及在SCF IL3 IL6 LF Tpo及SCF IL3 IL6 FL Tpo Ep组合下液体培养,其生存、增殖、扩增等能力上均无明显差异,说明CD34^ CD59^ 细胞在造血能力上并无内在的优势。GM—CSF或许是使PNH克隆呈造血优势的原因之一。  相似文献   

12.
为进一步探讨microRNA(miRNA)在造血调控中的作用奠定基础,比较了miRNA在人脐血CD34^+CD38^-、CD34^+CD38^+细胞中的差异性表达。从人脐血中分离单个核细胞(MNC),应用FACSVantage分选流式细胞仪分选CD34^+CD38^-、CD34^+CD38^+细胞;抽提miRNA后与miRNA芯片杂交,应用生物信息学技术分析miRNA芯片的表达结果。结果发现,miRNA在人脐血CD34^+CD38^+细胞的表达水平比在CD34^+CD38^-细胞的表达水平降低至1/2以下者共11个,表达水平升高至2倍以上者共73个,以上84个miRNA被称为“干细胞性”miRNA。经比较Georgantas等和芯片表达结果,发现有12个(14、29%,12/84)相同的miRNA。部分miRNA经历了类似于CD34在造血细胞表面的发展历程。应用生物信息学技术可寻找到新的与造血调控相关的miRNA簇及miRNA的下游靶基因。结论:“干细胞性”miRNA在正常造血中发挥着重要作用,即miRNA的系统表达模式一造血干/祖细胞基因表达谱一造血干/祖细胞的自我更新和系列定向分化。  相似文献   

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目的 为确定外周血CD34+细胞绝对计数能否可靠预示自体外周血干细胞的采集效果。方法 用流式细胞仪ProCOUNT方法对采集的 2 5份次移植物和采集当天外周血行CD34+细胞绝对计数 ,同时做外周血常规检查和移植物集落形成单位 (CFU)计数 ,每份次移植物以CD34+/kg ,单个核细胞 (MNC) /kg,粒 巨噬细胞集落形成单位 (CFU GM) /kg ,红细胞集落形成单位 (CFU E) /kg等为指标 ,与患者采集当天的外周血CD34+细胞绝对计数、CD34+细胞百分比、WBC ,MNC ,中性粒细胞(NEU)或血小板 (PLT)等各项指标进行相关分析和逐步回归分析。结果  ( 1)Spearman相关分析结果 :外周血CD34+细胞绝对计数与移植物CD34+/kg高度相关 (r=0 790 ,P <0 0 0 1) ,外周血CD34+细胞百分比与移植物CD34+/kg相关 (r=0 6 17,P <0 0 5 )。外周血WBC、MNC、NEU、PLT或RBC与移植物CD34+/kg无关。外周血CD34+细胞绝对计数与移植物CFU E相关 ,而与CFU GM无关。外周血MNC与移植物MNC/kg相关。 ( 2 )逐步回归分析结果 :移植物CD34+/kg只与外周血CD34+细胞绝对计数高度相关 (P <0 0 0 1) ,而与外周血CD34+细胞百分比无关。结论 移植物CD34+/kg只与外周血CD34+细胞绝对计数高度相关 ,外周血CD34+细胞绝对计数能够可靠预示自体外周血干细胞的采集效果  相似文献   

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BACKGROUND : Ex vivo expansion of HPCs is an attractive approach to overcoming the current limitations of human cord blood transplantation. It is important not only to define the optimal culture conditions but also to know the number of progenitor cells that can be obtained. CD34+ cells have a great variability in their cloning capacity and in their ability to expand HPCs. This study was carried out to assess whether this variability could be due to intrinsic or extrinsic factors.
STUDY DESIGN AND METHODS : CD34+ cells were analyzed for the expression of CD38, CD133, and CD117 and cultured in serum-free culture medium with four cytokine combinations: SCF plus thrombopoietin plus flt3 ligand (STF), STF plus IL-3, STF plus IL-6, and STF plus IL-6 plus IL-3. After a 1-week culture, the numbers of CD34+ cells and CFUs were determined.
RESULTS : The variability observed both in the cloning ability of CD34+ isolated cells and in their expansion capacity was inversely related to the frequency of the more immature CD34+CD38– cells. When more mature CD34+CD38+ cells were present within CD34+-isolated cells, a higher cloning ability, measured as CFUs, and a higher expansion capacity were observed.
CONCLUSION : Enumeration of CD34+CD38– cells is correlated with the number of committed progenitors and the capacity of generating CD34+ cells, an important parameter if expansion protocols must be used in clinical transplantation.  相似文献   

16.
Hematopoietic stem cells transplantation has been successfully used in the treatment of patients with hematological malignances. A better knowledge of the mechanisms beyond their ability to completely repopulate the entire hematopoietic system would help in the treatment of hematological diseases. For this reason we focused our studies on a cell population that has been demonstrated to have some peculiar characteristics among the stem cells: CD34+KDR+ cells. These cells, an extremely rare population among the CD34 (0.1%-0.5%) cells, have been demonstrated from different groups to have the potential to give rise to the hematopoietic and endothelial lineage. By a subtraction library approach we found different sequences more expressed in CD34+KDR+ than their CD34+KDR- counterpart. In particular, we found an open reading frame correspondent to a newly characterized E3 ligase, MARCH-I. This gene is part of a recently described family involved in immune response modulation through the proteosomal mediated degradation. MARCH-I expression in stem cells could be important for their intrinsic immune properties.  相似文献   

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Autologous hematopoietic stem cell transplantation (ASCT) is an established treatment for multiple myeloma (MM), yet the impact of transplanted CD34+ cell dose remains unresolved, especially in patients over the age of 65 years. Data was collected from 207 consecutive ASCT patients to determine the relationship between CD34+ infusion count and short-term and long-term platelet recovery. For MM patients under the age of 65 years (n = 155), CD34+ dosage correlates with time to platelet engraftment (p < 0.001) and platelet count at 30 days (p = 0.003), but not with long-term platelet counts at 180 or 360 days from the CD34+ reinfusion. For MM patients aged 65 years or older (n = 46), CD34+ dosage did not correlate with time to platelet engraftment, but did correlate with both short-term and long-term platelet counts at 30 (p < 0.001), 180 (p = 0.021), and 360 days (p = 0.005). Exploratory regression analysis was done to explore platelet stability following the current minimum CD34+ dosage reinfusion. For MM patients under the age of 65 years, the minimum standard CD34+ dosage of 2 × 106 cells/kg was sufficient for a timing to platelet engraftment of <21 days and short-term platelets count ≥150 × 109/L at 30 days. Alternatively, for MM patients aged 65 years or older, the CD34+ dosage of 2 × 106 cells/kg was insufficient for platelet counts ≥150 × 109/L at 30 and only marginally attainable at 360 days suggesting that in elderly MM patients a higher CD34+ dosage may be required for platelet recovery and possibly long-term platelet stability.  相似文献   

20.
BACKGROUND: Bone marrow-derived circulating progenitor cells (BM-CPCs) are mobilized into adult peripheral blood (PB) during acute myocardial infarction (AMI) and may contribute to the regeneration of infarcted myocardium. The purpose of the present study is to determine whether mobilization of BM-CPCs into PB depends on cardiovascular risk factors (CVRFs), age of patients, infarct associated inflammatory markers, and left ventricular function after AMI. MATERIALS AND METHODS: Peripheral blood concentrations of CD34/45(+) and CD133/45(+) BM-CPCs were measured by flow cytometry in 44 patients after AMI and in 16 subjects with atypical chest pain acting as controls. RESULTS: Mobilization of CD34/45(+) and CD133/45(+) BM-CPCs on day 1 after AMI showed significant negative correlation with age, the number of CVRFs, infarct size, creatine phosphokinase peak in bivariate as well as in multivariate analyses. We additionally found a positive correlation of CD34/45(+) and CD133/45(+) BM-CPCs mobilization on day 1 after AMI with global ejection fraction (EF) in bivariate analysis but could not confirm this in multivariate analysis. Elevated of C-reactive protein (CRP) and leukocyte levels on day 1 after AMI were significantly associated with decreased concentrations of CD34/45(+) BM-CPCs. The concentrations of CD34/45(+) and CD133/45(+) BM-CPCs significantly increased in AMI patients, with the peak on day 7 as compared to the control group. CONCLUSIONS: The mobilization of CD34/45(+) and CD133/45(+) BM-CPCs into the PB depends on many factors, i.e. the number of CVRFs, age, infarct size and inflammatory markers of patients. Most importantly, the severity of the circulatory dysfunction and the amount of necrotic myocardial tissue are the main determinants. Moreover, this spontaneous mobilization of BM-CPCs may serve as a very important surrogate for infarct size as well as for global EF and it may determine the regenerative potency after AMI.  相似文献   

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