脐血输注后再生障碍性贫血,白血病患者造血祖细胞体外培养及血 …

为探讨脐血输注的疗效机理，评价脐血输注的实用价值，我们用新鲜脐血静脉输注（ＵＣＢＴ），治疗慢性再生障碍性贫血（ＣＡＡ）１０例和急性白血病（ＡＬ）８例，观察ＵＣＢＴ前后患者粒单细胞系集形成单位（ＣＦＵ－ＧＭ）、红系爆式集落形成单位（ＢＦＵ－Ｅ）、集落刺激活性（ＣＳＡ）、爆式集落刺激活性（ＢＰＡ）及外周血象变化。结果提示：ＵＣＢＴ后可迅速升高ＣＦＵ－ＧＭ和ＢＦＵ－Ｅ；②可改善外周血象，尤其是ＷＢＣ、及     

脐血造血调节因子活性的研究

探讨脐血中造血调节因子含量，来源及其与造血细胞间间的关系。方法：双抗夹心酶联免疫肿附（ＥＬＩＳＡ）法和细胞因子依赖细胞株增殖反应ＭＴＴ法测定脐血和脐血细胞培养上清中颗粒细胞集落刺激因了（Ｇ－ＣＳＦ）、颗粒－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）、白细胞介素－６（ＩＬ－６）和肿瘤坏死因子－α（ＴＮＦα），并观察脐血造血调节因子与脐血（ＣＦＵ－ＧＭ、ＭＮＣ、ＷＢＣ相关关系。结果：正向造血调节因子Ｇ－ＣＳ     

HGFs联合对体外人脐血造血干细胞及SCID小鼠体内扩增作用的研究

本实验应用外源性重组人多种造血生长因子（ｒｈＨＧＦｓ）处理人ＨＳＣ／ＳＣＩＤ小鼠模型，以体外半固体培养法及ＦＣＡＳ检测ＣＦＵ－ＧＭ、ＣＦＵ－ＧＥＭＭ及ＣＤ３４＾＋细胞，从而评价ＨＧＦｓ对受鼠体内人造血干细胞的扩增作用。结果：人脐血及骨髓ＭＮＣ经孵育２４小时后ＣＤ３４＾＋细胞含量，ＣＦＵ－ＧＭ、ＣＦＵ－ＧＥＭＭ产率皆得到明显扩增（Ｐ〈０．０１）；且脐血组优于骨髓组（Ｐ〈０．０１）。输脐血ＭＮＣ＋ＨＧ     

应用细胞生长因子提高再生障碍性贫血患者造血干细胞质量的研究

为探讨再障发病机制中各种造血生长因子（ＨＧＦｓ）对其造血干细胞的影响,了解各种ＨＧＦｓ对造血干细胞生长、增殖的具体作用及相互间的协同作用,采用甲基纤维素半固体体外短期培养及长期液体培养的方法,观察ＨＧＦｓ如粒一巨噬细胞集落刺激因子（ＧＭ—ＣＳＦ）、粒细胞集落刺激因子（Ｇ—ＣＳＦ）、白细胞介素－３（ＩＬ—３）、ＩＬ—６、干细胞因子（ＳＣＦ）、促红细胞生成素（Ｅｐｏ）以及ＨＧＦｓ不同组合方式对再障患者骨髓造血干细胞增殖、分化的调控作用。发现再障患者的克隆形成细胞（ＣＦＣ）形成巨噬细胞粒细胞集落形成单位（ＣＦＵ—ＧＭ）和红系爆发式集落形成单位（ＢＦＵ—Ｅ）的能力受限,但大部分患者（１０／１２）骨髓中长期培养起始细胞（ＬＴＣ—ＩＣ）接近正常,提示再障的造血干细胞可能在分化成熟过程中受到阻碍所致。此外,各种ＨＧＦｓ均有不同程度的提高造血干细胞形成集落的能力,且不同ＨＧＦｓ之间对再障患者ＣＦＣ形成ＣＦＵ—ＧＭ、ＢＦＵ—Ｅ的能力同样具有协同作用,其中以ＳＣＦ、ＩＬ—３和ＣＭ—ＣＳＦ的联合应用更为显著（Ｐ＜０．００１）。     

FLt-3配基和促血小板生成素对脐血造血祖细胞的协同扩增作用

目的 探讨体外同时扩增粒／巨噬细胞系祖细胞（ＣＦＵ－ＧＭ）、巨核细胞系祖细胞（ＣＦＵ－ＭＫ）的最佳细胞因子组合。方法 利用免疫磁珠法分离经脐血ＣＤ３４细胞,在液体２体系中经干细胞因子（ＳＣＦ）白细胞介素（ＩＬ）－３、ＩＬ－６、粒系集落刺激因子（Ｇ－ＣＳＦ）、ＦＬｔ－３配基、促血小板生成素等不同细胞组合扩增１４ｄ,检测瓣微增倍数。结果 ＴＰＯ可协同ＳＣＦ、ＩＬ－３、ＩＬ－６、Ｇ－ＣＳＦ对ＣＦＵ－ＧＭ     

应用细胞生长因子提高再生障碍性贫血患者造血干细胞质量的研究

为探讨再障发病机制中各种造血生长因子（ＨＧＦｓ）对其造血干细胞的影响,了解各种ＨＧＦｓ对造血干细胞生长、增殖的具体作用相互间的协同作用,采用甲基纤维素半固体体外短期培养及长期液体培养的方法,观察ＨＧＦｓ如粒－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）、粒细胞集落刺激因子（Ｇ－ＣＳＦ）、白细胞介素－３（ＩＬ－３）、ＩＬ－６、干扰细胞因子（ＳＣＦ）、促红细胞生成素（Ｅｐｏ）以及ＨＧＦｓ不同组合方式对再障碍患     

儿童肿瘤强烈化疗联用rhG—CSF动员外周血造血干细胞

观察了７例（２例非何杰金ｉ淋巴瘤Ⅲ期、３例急淋、１例急粒及１例神经母细胞瘤Ⅳ期）儿童恶性肿瘤病人接受强烈化疗联用ｒｈＧ－ＣＳＦ后，外周血干细胞（ＰＢＳＣ）动员上升情况。ＰＢＳＣ上升的最明显特点是与外周血白细胞总数上升的幡度平行；ＷＢＣ的上升幅度与患儿骨髓是否受肿瘤细胞侵犯有关。本文显示ＣＤ３４＾＋细胞与粒单细胞集落形成单位（ＧＭ－ＣＦＵ）有较好的相关关系（ｒ＝０．８７，Ｐ〈０．００１），提示ＣＤ３     

造血干细胞移植治疗重型地中海贫血的临床进展

造血干细胞移植（ｈｅｍａｔｏｐｏｉｅｔｉｃｓｔｅｍｃｅｌｔｒａｎｓｐｌａｎｔａｔｉｏｎ,ＨＳＣＴ）是目前根治重型地中海贫血（简称地贫）的最佳方法。包括骨髓移植（ＢＭＴ）、脐血移植（ＵＣＢＴ）宫内造血干细胞移植、外周血造血干细胞移植（ＰＢＳＣＴ）。至今...     

粒－单细胞集落刺激因子在自体外周血干细胞移植中的临床应用

报告小儿自体外周血干细胞移植及化疗中粒。单细胞集落刺激因子（ＧＭ－ＣＳＦ）的临床应用研究，共２０例（２４次），并设对照组比较。结果：移植前用作动员剂９例。粒、单系祖细胞（ＣＦＵ－ＧＭ）产率提高平均２９倍，自体外周血每次采集的ＣＦＵ－ＧＭ数较对照组有显著提高（Ｐ＜０．０５）；用于移植后早期４例（６次），白细胞恢复天数较对照组明显缩短（Ｐ＜０．０５）；用于移植后中期及恢复期３例（４次），２例造血恢复，１例（２次）有短期疗效。用于化疗后白细胞减低者效果显著，副反应轻。认为ＧＭ－ＣＳＦ有明显动员外周血干细胞作用，用于移植后可促进造血功能恢复，提高移植的安全性。     

脐血血清对急性白血病化疗后骨髓抑制期CFU—GM生长的影响

探讨脐血血清对急性白血病化疗后骨髓抑制期造血细胞增殖的促进作用，采用体外ＣＦＵ－ＧＭ培养法观察正常成人血清和脐血血清对正常造血细胞和急性白血病化疗后骨髓抑制期造血细胞增殖的影响。结果显示：脐血血清能明显促进正常ＣＦＵ－ＧＭ和急性白血病化疗后骨髓抑制期ＣＦＵ－ＧＭ生长，因此：脐血在体外能够刺激正常骨髓和急性白血病化疗后骨髓抑制期骨髓造血祖细胞增殖，提示急性白血病强化疗和干细胞移植时输注脐血可促进骨髓     

慢性移植物抗宿主病的预防和治疗

目的　分析儿童异基因造血干细胞移植(allo-HSCT)后慢性移植物抗宿主病 (cGVHD)的特点及甲泼尼龙(MP)、吗替麦考酚酯(MMF)与他克莫司(FK506)或环孢素A(CSA)联合免疫抑制治疗的效果,探讨有效治疗方案。方法　共进行异基因造血干细胞移植 45例, 30例完全植入。完全植入的患儿中脐血移植(UCBT)17例,异基因外周血干细胞移植 (PBSCT) 13例。GVHD的预防采用CSA、MP、氨甲蝶呤及MMF等组合的方案。cGVHD患者采用MP+MMF+FK506或MMF+CSA联合治疗方案。结果　30例完全植入的患儿中发生cGVHD者 9例(30% );其中血缘相关PBSCT13例中发生cGVHD6例(46% );UCBT者完全植入 17例,发生cGVHD3例(18% )。急性GVHD迁延者 6例(67% )。1例患儿经CSA+MMF联合治疗痊愈; 8例采用MP+MMF+FK506“三联”治疗,cGVHD症状均得到有效控制,有效率为 100%, 2例分别并发巨细胞病毒间质性肺炎或败血症死亡,死亡率为22%; 7例无事件生存>3年(78% )。副作用有肝损害、肾损害、高血压、关节滑膜炎、心律失常,均在减药或停药、对症处理后毒副作用消失。感染是主要的合并症和死亡原因。结论　PBSCT者cGVHD发生率较UCBT者高;急性GVHD迁延是发生cGVHD的高危因素;选用包含MP、MMF、FK506 (或CSA)的 2~3种药物联合免疫治疗cGVHD,效果理想,患儿均能耐受,是     

人类巨细胞病毒感染对脐血造血祖细胞增殖的影响(英文)

目的：探讨人类巨细胞病毒(HCMV)感染对脐血造血祖细胞(CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk)体外增殖的抑制作用及其机制。方法：20例脐血标本收集于正常足月顺产新生儿。实验共分5组:(1)3个HCMV感染组,每个感染组分别加入0.1 mL的103、104及105空斑形成单位(PFU)HCMV-AD169病毒液于培养体系中;(2)灭活对照组,加入同体积灭活HCMV病毒液;(3)空白对照组,不加HCMV病毒液,代之以同体积的IMDM。采用造血祖细胞体外半固体培养技术,培养、观察、计数HCMV-AD169株对脐血CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk集落数、抑制率和集落维持时间;并用聚合酶链反应(PCR)技术检测集落细胞内HCMV-DNA。结果：(1)在造血祖细胞培养体系中加入不同滴度的HCMV-AD169后,104和105PFU滴度感染对CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk集落形成均有显著的抑制作用,103PFU滴度感染对CFU-Mix及CFU-Mk集落形成有显著的抑制作用,与空白对照组和灭活对照组比较,差异有显著性(P<0.05)。病毒滴度越高,抑制程度越明显(P<0.05)。(2)104和105PFU滴度感染组CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk集落维持时间较对照组明显缩短(P<0.01),103PFU滴度感染组CFU-Mix和CFU-Mk集落维持时间较对照组明显缩短(P<0.01)。(3)PCR显示3个感染组的CFU-GM、CFU-E、CFU-Mix及CFU-Mk集落细胞内均有HCMV-AD169DNA存在。结论：HCMV-AD169能直接感染CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk造血祖细胞,并抑制造血祖细胞的增殖,这可能与HCMV感染患儿出现粒细胞减少、血小板减少和贫血等造血功能紊乱有关。     

再生障碍性贫血患儿骨髓祖细胞培养的临床意义

目的探讨再生障碍性贫血(AA)患儿红细胞集落生成单位(CFU-E)、爆式红细胞集落生成单位(BFU-E)、粒细胞-单核细胞集落生成单位(CFU-GM)及巨核细胞集落生成单位(CFU-Meg)骨髓祖细胞培养情况,了解骨髓祖细胞在AA发病中的作用及意义。方法抽取28例AA患儿骨髓3～4mL,以淋巴细胞分离液提取单个核细胞,用磷酸盐缓冲液(PBS)洗涤2次,调节单个核细胞水平为106/mL,取0.3mL分别接种于红系、粒-单核系、巨核系培养基分别作4类祖细胞培养,在7、14d测定其集落数。结果28例AA中,4类集落数均值与对照组比较,差异有显著意义(P<0.01或P<0.001)。集落数随病情加重而逐步下降。4类集落数低于对照组患儿下限所占百分比分别为BFU-E35.71%、CFU-E85.71%、CFU-GM75.00%、CFU-Meg89.29%。治愈及进步者4类集落数接近对照组水平,治疗无效及进展者与治疗前比较无变化。结论动态监测AA患儿骨髓4类祖细胞集落数变化对辅助诊断、观察疗效及预后判断均有指导意义。     

Cryopreservation of Pluripotent and Committed Hemopoietic Progenitor Cells from Human Bone Marrow and Cord Blood

As one of the studies on autologous bone marrow transplantation for childhood cancer, the viabilities of several kinds of hemopoietic progenitor cells after cryopreservation was assayed using bone marrow and cord blood samples. The mean recovery rates of granulocyte-macrophage colony forming units (CFU-GM), erythroid burst forming units (BFU-E) and erythroid colony forming units (CFU-E) from bone marrow cells were 47.9%, 31.3% and 10.5%, respectively. The mean recovery rates of CFU-GM, BFU-E and CFU-E from cord blood cells were 46.7%, 39.1% and 22.4%, respectively. The ratio of primitive to mature BFU-E did not change after cryopreservation. The mean recovery rate of mixed colony forming units (CFU-mix) from marrow cells was 30.2%, and that from cord blood cells was 58.9%, demonstrating their sufficient proliferative activity after cryopreservation. These results demonstrate that CFU-mix, primitive BFU-E as well as CFU-GM, mature BFU-E can be well cryopreserved, supporting the usefulness of autologous marrow transplantation for the rescue of the ruined hemopoiesis after intensive cancer therapy.     

Effects of amifostine on clonogenic mesenchymal progenitors and hematopoietic progenitors exposed to radiation

PURPOSE: To determine the radiation sensitivities of mesenchymal progenitors and hematopoietic progenitors, and to determine the in vitro effects of amifostine on hematopoietic and mesenchymal progenitors exposed to radiation. METHODS: Radiosensitivity of mesenchymal progenitor cells was determined by exposing marrow low-density cells to radiation at doses of 100 to 800 cGy. Mesenchymal cell colonies were established by plating 2.5 x 10(5) marrow low-density cells in long-term marrow culture medium (LTCM). The size, frequency, and cellular composition of the mesenchymal progenitor cells were scored after 14 days of incubation. Mesenchymal progenitor cells were subdivided into progenitors forming fibroblast and adipocyte mixed colonies (CFU-FA), and pure fibroblast colonies (CFU-F). Hematopoietic progenitors were assessed by methylcellulose-based assay. RESULTS: Radiation at 100 cGy caused a mild decrease in CFU-F and CFU-FA derived colonies by 12% and 13%, respectively; 200 cGy decreased CFU-F by 36% and CFU-FA by 52%; 400 cGy decreased CFU-F by 50% and CFU-FA by 86%; and 600 cGy decreased CFU-F by 24%, with total absence of CFU-FA. Pretreatment with amifostine protected 100% of CFU-F at 100 and 200 cGy, 84% at 400 cGy, 46% at 600 cGy, and 14% at 800 cGy. With CFU-FA colonies amifostine pretreatment provided only minimal radioprotection. For hematopoietic progenitors radiation at 100 cGy reduced CFU-GM by 74% but had no significant effect on CFU-GEMM and BFU-E. Radiation at 200 cGy decreased CFU-GEMM by 72%, BFU-E by 54%, and CFU-GM by 84%; 400 cGy further decreased CFU-GEMM by 83%, BFU-E by 81%, and CFU-GM by 93%. Pretreatment with amifostine resulted in twofold stimulation of CFU-GEMM and BFU-E colonies. All BFU-E colonies were protected up to 200 cGy. For CFU-GEMM amifostine pretreatment resulting in 68% at 200 cGy and 31% at 400 cGy. For CFU-GM colonies it was 54% at 100 cGy, 32% at 200 cGy, and 12% at 400 cGy. CONCLUSIONS: Mesenchymal progenitor cell subpopulations are differentially sensitive to radiation. Amifostine protects both mesenchymal and hematopoietic progenitors against radiation injury, though the level of protection appears to be dependent upon the sensitivities of these progenitor cells to radiation. Amifostine is a potent stimulant of BFU-E and CFU-GEMM progenitor colonies.     

Chronic neutropenia: diagnostic approach and prognosis

Chronic neutropenia is a term used to describe a group of disorders characterized by a persistent neutrophil count of less than 1500 cells/microliters. We studied seven children and three sets of parents. We separated patients into a group with good prognosis and a group at higher risk of infection by using a combination of tests, including bone marrow aspiration and biopsy, steroid stimulation of bone marrow reserve, and in vitro CFU-GM and CSA assays. Children with a normal number of myeloid elements in their bone marrow and a normal bone marrow response to steroid stimulation had a benign course. CFU-GM and CSA assays helped to classify these children's neutropenia when their bone marrow had decreased numbers of myeloid elements. Family studies in three children were consistent with an inherited neutropenia, even when their parents were hematologically normal.     

川崎病并发冠状动脉病变的超声心动图表现的动态观察

目的 探讨川崎病(KD)所引起的冠状动脉扩张(CAD)以及冠状动脉瘤(CAA)的超声心动图特点.方法 用彩色多普勒超声心动图检测KD患儿冠状动脉开口和内径,并动态观察静脉免疫球蛋白治疗前后冠状动脉内径的变化情况.结果 本组46例KD患儿,41例患儿行超声心动图检查,合并CAD 12例,左冠状动脉(LCA)较右冠状动脉(RCA)更易受累及(P<0.05);CAA 4例,均为双侧冠状动脉病变,最大内径10 mm.治疗后6～18 d复查超声心动图,CAD组LCA较治疗前明显回缩(P<0.05),RCA内径无变化(P>0.05);CAA组LCA和RCA内径均无明显变化(P>0.05).对10例冠状动脉病变患儿进行随访,其中8例CAD冠状动脉内径均恢复正常,1例CAA冠状动脉病变加重,1例CAA完全恢复正常.结论 (1)在KD所引起的CAD中,LCA比RCA更易受累及,静脉免疫球蛋白治疗后短期内LCA明显回缩.(2)超声心动图是动态观察KD并发CAD和CAA的无创手段.     

A variant of the congenital dyserythropoietic anaemia type II with structural abnormalities in the granulocytic series

Typical features of congenital dyserythropoietic anaemia (CDA) were found in a 13-year-old girl admitted for chronic recurrent multifocal osteomyelitis. The findings on light microscopy were in agreement with those described in CDA type II, whereas on electron microscopy, the ultrastructure findings were compatible with both types I and II. The acidified serum lysis test (Ham test) performed with eight normal sera was negative. The patient's red blood cells showed an increased agglutinability with anti-i antibodies. Morphological changes were also shown in the mature neutrophilic granulocyte suggesting that the primary disorder exists already in the multipotent stem cell.Abbreviations CDA congenital dyserythropoietic anaemia - BFU-E erythroid burst-forming units - CFU-E erythroid colony-forming units - CFU-GM granulomonocytic colony-forming units - CSA colony-stimulating activity     

三羟异黄酮对雌二醇和双酚A促神经母细胞瘤细胞增殖的抑制作用

目的 观察大豆异黄酮类物质4',5,7,-三羟异黄酮(genistein,GEN)对17β-雌二醇(E2)和双酚A(BPA)诱导的神经母细胞瘤细胞增殖作用的影响.方法 SK-N-SH细胞经常规培养扩增后分为6组:组1,RPMI1640无酚红培养液加无水乙醇(对照组);组2,加GEN(12.5 μmol/L);组3,加E2(2μmol/L);组4,同时加E2和GEN;组5,加BPA(8 μmol/L);组6,同时加BPA和GEN.分别在24h、48h和72h进行光吸收度(A值)检测,72 h时用流式细胞仪检测细胞周期及凋亡亚二倍体峰;末端脱氧核糖核酸酶介导的dUTP末端标记(TUNEL)法检测细胞凋亡以及Westem blot检测磷酸化Akt和总Akt蛋白表达.结果 48 h和72 h时,组3、5与组1比较,细胞.A值有增加(P<0.01).48h和72h时组4与组3比较,组6与组5比较,细胞A值却降低(P<0.01).48 h时,组4的A值较组3降低10%,组6较组5降低13%(P<0.01);72h时.组4较组3的A值下降17%,组6较组5的A值下降22%(P<0.01).72 h时,组4的G2/M期细胞百分比是组3的1.78倍,组6是组5的1.49倍(P<0.01).72h时.各组磷酸化Akt表达有所不同,GEN干预组的磷酸化Akt光密度值较未干预组有下降.其中,组2的磷酸化Akt表达较对照组下降近15%;组4的磷酸化Akt表达分别较组3下降约11%;组6的磷酸化Akt表达较组5也有下降(均P<0.01);各组的总Akt蛋白表达未见明显差异.同样,各组的细胞凋亡未见统计学差异.结论 4',5,7,-三羟异黄酮可抑制17β-雌二醇和双酚A促神经母细胞瘤细胞体外增殖的作用,该作用可能与G2/M期细胞周期阻滞及PI3K/Akt信号转导通路异常有关.     

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??Objective??To observe the efficacy and safety of multi-target immunosuppressive therapy in treatment of children with steroid-resistant nephritic syndrome??SRNS??. Methods??A total of 48 children with SRNS were enrolled in this multicenter prospective study. Based on the same comprehensive treatment??the children were randomly divided into two groups?? ??1??observation group?? they were orally given CSA??3??4 mg/??kg·d???? and MMF??20 mg/??kg·d???? multi-target immunosuppressive therapy????2??control group??they were orally given CSA??4??6 mg/??kg·d????. The side effects were closely observed. The plasma concentrations of CSA??urine protein to creatinine ratios??liver and kidney function??blood routine and urine β2-microglobulin were respectively compared between the two groups after 2 weeks??1 month??3 months and 6 months of treatment. Results??The average plasma concentrations of CSA in the observation group was ??88.86±16.94?? μg/L??and in the control group it was ??152.96±19.20?? μg/L??P??0.001??. The urine protein to creatinine ratios in the observation group after 1 month and 3 months of treatment were lower than the control group in the same time period??P??0.05??. The serum albumin in the observation group after 1 month and 3 months of treatment was higher than the control group in the same time period??P??0.05??. The urine β2-microglobulin in these two groups had no differences during the treatment??P??0.05??. The overall remission rate of the observation group was 88%??and in the control group it was 87%. The remission rate of the observation group after 2 weeks and 1 month of treatment was better than the control group??P??0.05??. The main side effect during therapy was infection??gastrointestinal reaction??crinosity??hypertension and leukocyte decrease. The side effect of the observation group was less than the control group??P??0.05??. Conclusion??The multi-target therapy in children with SRNS by CSA and MMF results in early remission and can keep long-term remission with mild side effect.