Histopathology of bone marrow reconstitution after allogeneic bone marrow transplantation

In order to study haematopoietic reconstitution in allogeneic bone marrow transplantation we investigated bone marrow histology in 61 biopsies of 37 patients, treated with HLA-compatible bone marrow grafts for leukaemia or severe aplastic anaemia. The biopsies were taken from the day of transplantation until 100 d after transplantation. Stromal changes, in particular oedema, fibrosis and granulomas, were found during the whole period of observation. These changes were more prominent in biopsies from leukaemia patients than from patients with aplastic anaemia. The cellularity in the biopsies increased until 28 d after bone marrow transplantation and was stable thereafter. Initially, only clusters of cells belonging to a single cell lineage were seen, suggesting that the first outgrowth of haematopoietic cells is by proliferation of committed precursor cells. Long-lasting abnormalities in localization of haematopoietic cells in the bone marrow space and of the myeloid: erythroid ratio were seen; dyserythropoiesis was common.     

人脐血间充质干细胞移植修复骨髓造血损伤

文题释义：间充质干细胞：是一种多功能干细胞,由胚胎发育早期的中胚层发展而来,存在于人的各种组织、器官中,如骨、软骨、脂肪、外周血和肌肉等。骨髓组织中的间充质干细胞最多,但其在骨髓细胞中的比例仍很低,只有0.01%-1.00%,且年龄越大其含量越少,骨髓中的间充质干细胞分化能力也呈下降趋势。与骨髓相比,脐血中所含的间充质干细胞更加原始,因而有更强的增殖、分化能力。相较于骨髓间充质干细胞而言,人脐血间充质干细胞具有来源广泛、取材方便、无伦理方面的限制,使得人脐血间充质干细胞成为再生医学中的另一重要来源。骨髓造血损伤动物模型：建立骨髓造血损伤动物模型的方法有很多,主要分为物理方法、化学方法及物理化学方法等。物理方法包括各种射线,如X射线等,化学方法主要指以环磷酰胺为代表的烷化剂类化疗药物,而物理化学方法也叫混合性方法,联合应用放射线和化疗药物建立动物模型。  摘要背景：大多数研究间充质干细胞体外培养对造血干细胞的增殖作用和骨髓间充质干细胞移植可降低辐照引起的造血细胞死亡,增加骨髓细胞存活,修复造血功能,而少有研究人脐血间充质干细胞移植对骨髓造血损伤的修复。目的：探讨人脐血间充质干细胞对骨髓造血微环境的修复情况。方法：选用雄性BALB/c小鼠随机分为3组,实验组和对照组小鼠进行总剂量为6 Gy的X射线全身照射,建立骨髓造血损伤模型,正常组为未经处理的正常小鼠。实验组小鼠照射当天经尾静脉输入CM-DiL标记的人脐血间充质干细胞5×10⁶/只(0.2 mL),对照组和正常组经尾静脉输入生理盐水0.2 mL,移植后第1,5,7,14,21天观察外周血血象恢复情况和骨髓造血微环境修复情况。结果与结论：①外周血常规：移植后第1,5,7天,实验组和对照组小鼠与正常组小鼠比较,白细胞、血小板、红细胞计数及血红蛋白浓度进行性下降,第7天下降最为明显,移植后第14天三系较前有所恢复,移植后第21天基本恢复正常,与实验组相比,对照组三系下降更为明显,移植后第14天实验组较对照组恢复快;②骨髓涂片情况：移植后第1,5,7,14天实验组及对照组小鼠骨髓出现造血功能抑制,以第7天最为明显,移植后第14天骨髓增生较前有所恢复,实验组优于对照组;移植后第21天实验组及对照组小鼠骨髓造血功能恢复,与正常组相比无差异;③骨髓病理切片情况：移植后第1,5,7,14天实验组及对照组小鼠骨髓出现造血功能抑制;移植后第14天实验组及对照组小鼠的骨髓造血功能较前开始恢复,实验组小鼠的骨髓增生情况优于对照组小鼠, 移植后第21天实验组及对照组小鼠骨髓增生情况与正常组比较无差异;④结果表明,人脐血间充质干细胞对骨髓造血功能恢复均有明显促进作用。ORCID: 0000-0002-7547-9664(高坤莉) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

同胞与非血缘脐血移植治疗儿童恶性血液病的效果分析

背景:近年来,脐血逐渐成为亲缘及非血缘骨髓或外周血造血干细胞移植的一种极其关键的替代干细胞来源,被越来越多地用于儿童恶性血液病的治疗。目的:比较同胞与非血缘脐血移植治疗儿童恶性血液病的临床疗效。方法:回顾性分析1998-01-01/2018-12-31于郑州大学第一附属医院造血干细胞移植中心接受同胞脐血移植及非血缘脐血移植治疗儿童恶性血液病患者的临床资料,所有脐血移植患者均采用清髓性预处理方案,同时应用环孢素A±吗替麦考酚酯方案预防移植物抗宿主病。结果与结论:①2例同胞脐血移植患者及3例非血缘脐血移植患者造血植入失败继发感染死亡,其他全部脐血移植患者均顺利达到造血植入;同胞脐血移植组、非血缘脐血移植组中性粒细胞与血小板中位植入时间分别为[17 d(11-43 d),18 d(12-45 d),P=0.307]与[20.5 d(15-50 d),27 d(18-56 d),P=0.773],差异均无显著性意义;②同胞脐血移植组、非血缘脐血移植组急性移植物抗宿主病与慢性移植物抗宿主病的发生率分别为(36%vs.43%,P=0.737)与(15%vs.33%,P=0.412),差异均无显著性意义;同胞脐血移植组与非血缘脐血移植组移植后感染的发生率为56%,71%,差异无显著性意义(P=0.343);③同胞脐血移植组、非血缘脐血移植组2年总体生存率与2年无复发生存率分别为(61%vs.36%,P=0.301)与(56%vs.33%,P=0.151),差异均无显著性意义;同胞脐血移植组、非血缘脐血移植组5年总体生存率和5年无复发生存率分别为(54%vs.24%,P=0.044)与(50%vs.20%,P=0.039),两组在长期生存方面差异有显著性意义;④结果显示同胞与非血缘脐血移植均是治疗儿童恶性血液病安全有效可行的移植方式,尤其在儿童血液病患者替代供者移植的长期生存方面明显受益。     

骨髓腔内途径脐血与间质干细胞共移植对造血重建的实验研究

目的：探讨骨髓腔内输注（IBM）脐血与间质干细胞（MSCs）对大鼠造血重建、骨髓MSCs恢复的影响，并研究供体MSCs植入状态以探讨MSCs的作用机制。方法：BrdU标记F344大鼠骨髓MSCs通过双侧胫骨IBM或尾静脉注射（IV）与胎鼠及新生大鼠外周血（FNPB）共移植Wistar雌鼠。监测受鼠存活状况、造血免疫重建、HSCs植入水平及骨髓MSCs恢复情况，并以免疫荧光法检测受鼠骨髓MSCs的来源。结果：(1)2个共移植组60 d存活率均为100%，单纯FNPB移植组仅为66.7%。(2)共移植组的外周血象、骨髓造血干祖细胞集落产率明显高于单纯FNPB移植组，尤以骨髓腔共移植组最佳。(3)2个共移植组的HSCs植入水平无统计学差异，而骨髓腔共移植组明显高于单纯FNPB移植组（P＜0.05）。(4)30 d时各移植组MSCs的增殖能力未达正常水平，但仍以骨髓腔共移植组的恢复情况最佳（P＜0.05）。(5)仅少部分受体可发现供、受体源性MSCs嵌合。 结论：脐血与MSCs共移植可促进受体骨髓MSCs恢复和造血重建，提高HSCs植入率；IBM途径应用安全，促进造血恢复的作用优于IV途径。     

Immunologic reconstitution following bone marrow transplantation for X-linked hyper IgM syndrome

X-linked hyper IgM syndrome (XHIM), caused by mutations of the CD40 ligand (CD40L) gene, is characterized by recurrent bacterial and opportunistic infections, an increased incidence of autoimmunity and malignancies, and immunodeficiency due to abnormal T/B cell interaction. Because of poor long-term prognosis, bone marrow transplantation (BMT) has been proposed as an alternative treatment. An 8-month-old boy with XHIM and a splice site mutation of CD40L underwent BMT using a fully matched sibling donor. Markers of engraftment and immunologic reconstitution were measured serially. After BMT, activated T cells expressed functional CD40L, and genomic DNA obtained from circulating white cells contained predominantly wild-type CD40L sequences. Serum immunoglobulin levels including IgE and antibody responses to recall antigens normalized, and immunization with the T-cell-dependent neoantigen, bacteriophage φX174, demonstrated amplification of the response and isotope switching. BMT provides a permanent cure for XHIM if a fully matched sibling donor is available and the procedure is performed before complications have occurred.     

脐血与成人骨髓淋巴细胞及其亚群比较

比较脐血与骨髓T淋巴细胞、B淋巴细胞和NK细胞亚群的分布特点及血清sIL-2R和IgG水平的不同,探讨脐血移植后免疫重建延迟、GVHD及GVL效应的发生机制。应用流式细胞仪分别检测脐血及骨髓CD3+、CD3+CD5+、CD3+CD45RA+、CD3+CD45RO+、CD3+CD25+、CD19+、CD19+CD10+、CD19+CD40+、CD19+CD23+、CD3-CD16+CD56-及CD3-CD16+CD56+细胞的含量。应用双抗夹心酶联免疫吸附法(ELISA)及速率免疫散射比浊法分别检测脐血及骨髓血清中sIL-2R及IgG水平。CD3+T细胞数在脐血及骨髓中无显著差异(P>0.05)。脐血中CD3+CD45RA+、CD19+、CD19+CD10+及CD3-CD16+CD56-细胞数显著高于骨髓(P<0.05)。脐血中CD3+CD5+、CD3+CD45RO+、CD3+CD25+、CD19+CD40+、CD19+CD23+及CD3-CD16+CD56+细胞数显著低于骨髓(P<0.05)。脐血血清中sIL-2R及IgG水平显著低于骨髓(P<0.05)。脐血中成熟T细胞、成熟B细胞及成熟NK细胞亚群的细胞数显著低于骨髓,这可能是脐血移植后免疫重建延迟、GVHD发生率低及GVL效应低的原因之一。     

不同培养体系对脐带血造血干细胞扩增的影响

目的探讨不同培养体系对造血干细胞的体外扩增及其表型的改变。方法新鲜分离人脐带血单个核细胞（MNC），免疫磁珠法分选CD34^＋造血干细胞（HSC），计数富集得到的CD34^＋细胞，平均分为3组，每组含CD34^＋细胞2．2×10^5：A组（HSC＋CK）CD34^＋细胞接种于Stemline^TMⅡ无血清培养基中．加入早期作用因子FST组合（SCF、FL和TPO，质量浓度50ng／ml的SCF、质量浓度100ng/ml的TPO和FL），并于接种0d添加质量浓度20ng／mlIL-3：B组（HSC＋MSC）CD34^＋细胞接种于含MSC feeder的培养瓶．加入Stemline^TMⅡ无血清培养基：C组（HSC＋MSC＋CK）CD34^＋细胞接种于含MSC feeder的培养瓶．加入Stemline^TMⅡ无血清培养基，加入早期作用因子FST组合首剂添加IL-3（剂量同A组）。在培养后4、7、10、14d计数有核细胞总数，流式细胞术检测扩增细胞免疫表型的改变。结果0～14d培养后MNC细胞扩增数C组（HSC＋MSC＋CK）〉A组（HSC＋CK）〉B组（HSC＋MSC），P〈0．01。3组间CD34^＋细胞比例B组（HSC＋MSC）〉C组（HSC＋MSC＋CK）〉A组（HSC＋CK），P〈0．01。CD34^＋细胞绝对数也出现了明显增加，其中C组（HSC＋MSC＋CK）增加最为明显．其次是A组（HSC＋CK）。其中A组（HSC＋CK）培养4d较0d（14．68％）CD34^＋CD38^-细胞有明显增加（62．71％，P〈0．05），C组（HSC＋MSC＋CK）CD34^＋CD38^-细胞也略有增加（23．99％）：培养7d时，A组（HSC＋CK）、C组（HSC＋MSC＋CK）CD34^＋CD38^-细胞数明显下降，分别为4．44％和1.38％，而B组（HSC＋MSC）CD34^＋CD38^-细胞上升为18．92％，与0d时比较P〈0．05，与A组（HSC＋CK）、C组（HSC＋MSC＋CK）比较P〈0．05。结论MSC和细胞因子的联合应用，一方面使得总MNC细胞得到大量扩增，同时还使扩增后细胞保持CD34^＋免疫表型，培养体系中加入MSC能更有效／特异地扩增CD34^＋造虹干细胞群.     

华北地区1026例脐带血细胞各参数结果分析

目的确定华北地区脐带血血细胞和造血祖细胞参数及其相关性。方法采集1026例符合标准的脐带血标本,分别用全自动五分类血细胞计数仪进行脐带血细胞分类计数,流式细胞术和半固体培养进行造血干/祖细胞特性检测。结果脐带血中RBC穴4.06±0.47雪×1012/L,HGB穴149.0±18.2雪g/L,Plt穴241.2±53.7雪×109/L,WBC穴13.6±4.6雪×109/L,NEUT穴5.16±3.2雪×109/L,LYMP穴4.53±1.97雪×109/L,MONO穴0.78±0.36雪×109/L,NRBC穴0.45±0.45雪×109/L,RET穴0.14±0.04雪×1012/L。CD34阳性率0.36%±0.22%,CFU-GM和BFU-E产率(/5×104)分别为23.2±11.4和31.1±17.2。结论脐带血由独特的细胞组成,其血细胞各项参数多高于成人静脉血,其造血细胞具有更强的增殖潜力。     

人骨骼肌源性血管外膜细胞对人脐血CD34+细胞的体外造血支持作用#br#

文题释义： 血管外膜细胞：是血管周围星状细胞,分布于全身的毛细血管和微血管的管壁,是血管周围微环境的重要核心组成成分。它们表达CD146、NG2、PDGFRβ、LepR、Nestin等标记物,而不表达内皮细胞标记物CD144、vWF、CD31及造血细胞标记物CD34、CD45、CD14。研究显示血管外膜细胞是间充质干细胞的前体细胞,其表达间充质干细胞表面标记物,具有多向分化潜能,并可支持造血。 造血干细胞微环境：是造血组织中造血干细胞赖以生存并进行自我更新、多向分化的场所。它由骨内膜微环境和血管微环境组成,前者维持造血干细胞的静止及自我更新,后者促进造血干细胞动员、增殖、分化。 背景：研究显示血管外膜细胞是间充质干细胞的前体细胞,其通过细胞接触或旁分泌效应调节造血干细胞的行为并支持造血,人骨骼肌源性血管外膜细胞对造血的支持作用有待于研究。 目的：从人骨骼肌中分离培养血管外膜细胞并进行生物学特性鉴定,研究其对脐血CD34⁺细胞的体外支持作用。 方法：①利用多参数流式细胞术从人骨骼肌中分选表型为CD146⁺CD56^-CD34^-CD144^-CD45^-的血管外膜细胞,并对其进行生物学鉴定;②建立以CD146⁺人骨骼肌源性血管外膜细胞为滋养层的脐血CD34⁺细胞体外培养体系(实验组),以人骨髓间充质干细胞为滋养层的脐血CD34⁺细胞体外培养体系为阳性对照组,共培养1,2,4周检测培养体系的细胞数量、集落形成能力及免疫表型并进行统计分析。 结果与结论：①通过多参数流式细胞术分选出CD146⁺人骨骼肌源性血管外膜细胞,回测纯度为(91.5±1.85)% (n=5);其表达间充质干细胞表面抗原CD73、CD90、CD105、CD44,不表达造血细胞及内皮细胞表面抗原CD45、CD34、CD31;经诱导培养可向骨细胞、软骨细胞、脂肪细胞及肌细胞分化;②实验组与阳性对照组相比,在细胞数、集落形成能力及免疫表型方面(CD45⁺、CD34⁺CD33^-、CD14⁺、CD10⁺/CD19)差异均无显著性意义(P > 0.05,n=6);无滋养层的空白对照组培养1周时细胞数量明显减少,2周时几乎无细胞存活;③结果表明,CD146⁺人骨骼肌源性血管外膜细胞与人骨髓间充质干细胞一样对脐血CD34⁺细胞具有体外支持作用。 ORCID: 0000-0002-6768-5273(郑波) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

脐血干细胞移植治疗假肥大型肌营养不良症

目的比较假肥大型肌营养不良症(Duchennemusculardystrophy,DMD)患者经脐血干细胞移植治疗前后其肌肉再生、抗肌萎缩蛋白表达和运动功能的改变;以及评价治疗的安全性。方法对1例经基因分析和肌肉活检及抗肌萎缩蛋白检测确诊的、已丧失行走能力的DMD患儿,经HLA配型,在脐血库中寻找到一个全相合的脐血供体。采用白消安+环磷酰胺+兔抗胸腺淋巴细胞球蛋白预处理后进行异基因脐血干细胞移植;术后采用环孢素A和骁悉方案预防移植物抗宿主反应(graftversushostreaction,GVHD)。同时定期检测原发病的生化指标如血清肌酸激酶(creatinekinase,CK)、造血重建的植入证据(血型转变、肌肉和血液系统的聚合酶链反应短串联重复序列分析)、缺陷基因是否纠正、新生肌肉是否出现、肌肉中抗肌萎缩蛋白是否表达和运动功能是否改善。结果(1)中性粒细胞在脐血干细胞移植后第15天(+15天)达到0.5×109/L,白细胞在+25天达正常水平;血小板于+22天达到20×109/L;血红蛋白维持于85～100g/L。术后140天骨髓穿刺提示三系生长旺盛;(2)移植后140天血型转为供体AB型。至今没有出现移植物抗宿主反应。(3)术后18天、30天、43天、55天、74天、233天患者外周血DNA和术后140天、183天、235天骨髓细胞DNA经PCRSTR检测为供者独立植入;(4)患儿术后60天取外周血做基因分析,显示19号缺失的外显子得到完全纠正,患儿转变为正常基因型;(5)患儿在移植后75天的肌肉活检可见新生肌管形成,抗肌萎缩蛋白免疫组化呈弱阳性,少数为强阳性反应,DNA分析:供者基因DNA占1%～13%;移植后126天抗肌萎缩蛋白免疫组化检测显示阳性的肌纤维明显增多,供者基因DNA上升至2.5%～25%;(6)患儿血清CK从移植治疗前的5735U/L降至274U/L;(7)术后100天体检发现患儿肌力略有改善,肢端温暖。结论异基因脐血干细胞移植治疗DMD,可在移植后短期内重建造血功能、血清CK显著下降、肌肉抗肌萎缩蛋白表达,患儿运动有所改善,提示造血干细胞移植将有益于DMD的治疗。     

IgG subclass levels and immune reconstitution after T cell-depleted allogeneic bone marrow transplantation

Defects of humoral immunity are well documented after bone marrow transplantation (BMT). Immunoglobulin recovery can be impaired and selective deficiencies of IgG subclasses have been reported. The nature of these deficiencies may reflect patterns of infection in the post-BMT period. We studied immunoglobulin and IgG subclass recovery in 20 long term (greater than 100 days) survivors of T depleted allogeneic BMT. Although there was no fall in mean levels of IgG, IgM or IgA for the patient group, 14 patients (70%) developed a deficiency of one or more immunoglobulin isotype at some stage post-BMT. Eight patients (40%) had deficiency of IgG, IgA and IgM and six had selective deficiencies. When IgG subclasses were measured it was seen that mean levels of IgG2 and IgG4 fell post-BMT with trough levels occurring at around 120 days post-transplant. Sixty per cent of patients developed IgG2 subclass deficiency and of these patients 78% had an associated IgG4 deficiency. Deficiencies of IgG1 and IgG3 were less common and less prolonged than those of IgG2 and IgG4; in addition, mean levels of IgG1 and IgG3 showed a rise early post-BMT. In conclusion, a majority of our patients developed immunoparesis following BMT, usually at around 120 days after transplantation. IgG2 subclass deficiency, often in association with IgG4 deficiency, is common and may occur despite normal total IgG levels. Deficiencies of immunoglobulin and IgG subclasses may persist for longer than 1 year post-BMT. Differing profiles of immunoglobulin and IgG subclass recovery may help dictate patterns of infection in long-term survivors of BMT.     

早期作用造血因子对人脐血CD34^＋细胞的体外扩增作用

为了观察早期作用造血细胞因子ＳＣＦ、ＦＬ、ＩＬ－３、ＩＬ－６、ＴＰＯ单独及联合应用,对脐血ＣＤ３４＋细胞的体外扩增作用。我们用吸附单克隆抗体一磁珠分离系统富集人脐血ＣＤ３４＋细胞,在体外液体培养体系中加入不同的细胞因子扩增４周,每周取样计数有核细胞总数及集落形成细胞（ＣＦＣ）数。结果表明：用磁性细胞分离议富集脐血ＣＤ３４＋细胞纯度为８０％～８７％;一些细胞因子有明显的协同效应,其联合应用的扩增作用显著高于单因于作用：ＳＣＦ＋ＦＬ存在下,ＩＬ－３是有效扩增有核细胞总数及ＣＦＣ的关键因子了;细胞因子ＳＣＦ＋ＦＬ＋ＩＬ－３和ＳＣＦ＋ＦＬ＋ＩＬ－３＋ＩＬ－６组合对有核细胞总数及ＣＦＣ均有良好的扩增效应,培养２周时对ＣＦＣ的扩增倍数分别为３８．３±４．４和２９．６±２．７倍,可满足成人移植及基因治疗等的需要。     

脐血细胞神经分化与移植的研究进展

自20年前Nakahata等发现人脐血中含有丰富的造血干细胞以来,脐血造血干,祖细胞的研究取得了很大的进展,并已成为一个新的造血干细胞来源。近来研究表明,脐血中的部分细胞在体外培养或体内移植后可分化为神经细胞,并可促进受损动物的神经功能恢复,本文将对脐血源神经细胞的研究进展进行探讨。     

Morphology of the bone marrow after stem cell transplantation

In many haematological conditions the only curative option is stem cell (SCT) or bone marrow (BM) transplantation. Little information exists about BM morphology following non-ablative engraftment. During the pretransplantation period and depending on the kind of pretreatment, there may be hypoplasia, residual disease and varying degrees of fibrosis. In the post-transplantation period, after 1-3 weeks of transfusion-dependent pancytopenia, the first signs of successful engraftment are indicated by the recurrence of neutrophils, monocytes and erythrocytes in the peripheral blood. In the BM there is slow regeneration of erythropoiesis, followed by the other lineages of haematopoiesis and increase in reticulin fibres or even a resolution of fibrosis. Diagnostic problems arise when neoplastic lympho- or haematopoiesis are maintained following transplantation. Moreover, there may be a significant graft versus tumour response reaction or an already relapsing disease needing aggressive treatment. On the other hand, a conspicuous dyshaematopoiesis should not be mistaken as representing a myelodysplastic syndrome. The presence of granulomas being treatment-related or a manifestation of intercurrent granulomatous disease has to be considered. More advanced knowledge of the histological features of regenerating BM will certainly aid the recognition of relapsing disease and is needed for the adequate reporting of post-transplant alterations associated with a successful or failing engraftment.     

脐带血间充质干细胞的研究进展

脐带血中存在着丰富的造血干细胞,在移植方面发挥重要作用,在脐血中是否还存在间充质干细胞却有争议,有的学者认为其中有间充质干细胞,且与骨髓间充质干细胞(mesenchymalstemcells,MSC)的形态、表面标志及分化潜能非常相似,有的学者认为含量较低,难以传代培养扩增,总结了近5年来国内外关于脐血间充质干细胞的研究情况,为脐血的充分利用提供更多的资料。     

人脐血MSCs移植修复大鼠脊髓损伤的效果及机制的初步探讨

目的　探讨人脐血间充质干细胞（MSCs）移植修复大鼠脊髓损伤的作用及机制。 方法　分离纯化人脐血MSCs;制备大鼠脊髓半横断损伤模型,随机分为三组,分别在术后3 d经尾静脉注射生理盐水、培养液和BrdU标记的MSCs。移植后7、14、21、28 d,采用BBB评分法评估各组大鼠脊髓功能恢复情况;免疫荧光双标法检测MSCs在脊髓内的迁移、存活和分化,免疫组织化学法检测炎症因子高迁移率族蛋白B1（HMGB1）和核因子（NF-κB）在脊髓损伤部位的表达规律。 结果　移植后28 d,MSCs移植组大鼠肢体功能恢复明显,与生理盐水组和培养液组比较差别有统计学意义(P＜0.05)。移植后7、14、21d,脊髓损伤区及周边均可见大量Brdu+细胞,其中BrdU+GFAP+细胞约占53.3%,BrdU+NSE+细胞约占　22.15%。相同时间点MSCs移植组HMGB1和NF-κB的阳性表达率远低于生理盐水组和培养液组,差别有统计学意义(P＜0.05)。 结论　人脐血MSCs移植后可替代损伤的神经细胞,并可减轻脊髓损伤后的炎症反应,从而促进大鼠脊髓功能的恢复。     

Application of autologous bone marrow stem cells in the therapy of spinal cord injury patients

We studied the safety and efficiency of transplantation of autologous bone marrow cells in complex therapy of patients with spinal cord injury in the late period of the disease. In control group patients, meningomyeloradiculolis was performed, while in the main group surgical treatment was supplemented by transplantation of autologous bone marrow cells. Transplantation of BM stem cells into the cyst cavity and intravenously was well tolerated, did not cause allergic or inflammatory reactions in the early and delayed periods after surgery, and did not induce the formation of ossification foci in the nervous tissue. Analysis of the neurological status by ASIA, Bartel, and Ashworth scales showed that in the main group the positive clinical dynamics was more often observed than in the control. The decrease in neurological deficit included improvement of sensory and motor activity and conducting sensory function. Thus, transplantation of autologous bone marrow cells can be a novel safe strategy for the treatment of patients in the late period after spinal trauma. __________ Translated from Kletochnye Tehnologii v Biologii i Medicine, No. 2, pp. 109–114, April, 2007     

Comparison of two methods used for monitoring low-copy cytomegalovirus infection in a patient with chronic myeloid leukemia after unrelated umbilical cord blood transplantation

Introduction:

Detection of human cytomegalovirus (CMV, HHV-5) DNA in clinical specimens is considered a cornerstone in the diagnosis of HHV-5 disease. The present study compared two quantitative methods used for diagnosing cytomegalovirus infection in a 21-year-old woman with chronic myeloid leukemia after an unrelated umbilical cord blood transplantation.

Materials and Methods:

Blood samples were tested for the presence of HHV-5 DNA using the LightCycler PCR, the quantitative Eclipse^® CMV DNA Detection Kit, and a qualitative in-house PCR assay using primers that amplify part of the HHV-5 MIE gene.

Results:

Results from samples containing a low cytomegalovirus load were more accurate with the LightCycler test than those obtained with the Eclipse^® test, which underestimated the viral load of samples containing low DNA copy numbers.

Conclusions:

These findings underline the value of novel PCR methods used in current therapeutic procedures and in monitoring antiviral therapy with nucleoside analogs. The high level of sensitivity, specificity, accuracy, and rapidity provided by the LightCycler instrument are favorable for the use of this system in the detection of HHV-5 DNA in clinical specimens.

     

Functional analysis of CD8 lymphocytes in long-term surviving patients after bone marrow transplantation

The recovery of T-cell populations after bone marrow transplantation (BMT) is characterized by a persistent expansion of CD8 lymphocytes. Previously, we have shown that beyond 1 year posttransplantation the CD8 lymphocytes consist, to a large extent, of CD8⁺ HNK1⁺ cells that suppress, like normal CD8 lymphocytes, immunoglobulin productionin vitro. We have further investigated the functional capabilities of CD8 lymphocytes, mostly HNK1⁺ (from 50 to 77%), in seven long-term BMT patients. As normal, patient CD8 lymphocytes do not suppress (1) phytohemagglutinin (PHA)-induced interleukin 2 (IL2) receptor expression and IL2 responsiveness by normal T cells or (2) the mixed lymphocyte reaction of donor cells. Also as normal, patient CD8 lymphocytes can be activated into potent cytotoxic effectors. Therefore, under the present experimental conditions, the increase in the absolute number of CD8 lymphocytes in the long-term BMT patients is characterized by an expansion of the CD8⁺ HNK1⁺-cell subpopulation and a normal suppressor/cytotoxic potential on a per-CD8⁺ cell basis.     

Intractable diseases and bone marrow transplantation

Intractable diseases are defined as diseases of unknown etiopathogenesis, and for which therapeutic strategies remain to be established. They are therefore likely to cause various sequelae. Of the 86 intractable diseases recognized by the Ministry of Health and Welfare of Japan, half are thought to be curable by bone marrow transplantation (BMT).
This report shows which diseases are curable by BMT and provides evidence that autoimmune diseases are stem cell disorders.