骨髓干细胞移植促进ctnt^R141W转基因小鼠心肌组织再生

目的观察骨髓干细胞移植对ctnt^R141W转基因小鼠心脏组织结构的重建和病变心脏的泵血功能的影响。方法放射线9Gy照射ctnt^R141W转基因小鼠,将绿色荧光蛋白转基因小鼠的骨髓细胞尾静脉注射入ctnt^R141W转基因小鼠体内,替换ctnt^R141W转基因小鼠自身的骨髓细胞。流式细胞仪分析移植后的ctnt^R141W转基因小鼠绿色荧光骨髓干细胞所占的百分比。用免疫荧光双染方法探测移植后的ctnt^R141W转基因小鼠心肌组织中绿色荧光心肌细胞所占的比例,M型超声检测ctnt^R141W转基因小鼠心脏功能变化。结果相对于野生型对照小鼠ctnt^R141W转基因小鼠心肌组织中绿色荧光心肌细胞显著增加,左心室内径缩小,室壁变厚,收缩期末期和舒张末期左室容积缩小,心脏泵血功能增强。结论移植后的骨髓干细胞参与ctnt^R141W转基因小鼠心肌组织的损伤后重建,有助于改善病变心脏的泵血功能。研究提示扩张型心肌病心肌组织损伤重建增加,移植后的骨髓干细胞在受损心肌组织再生过程中起重要作用。     

Bone marrow hypoplasia complicating tacrolimus (FK506) therapy

Tacrolimus (FK506)-induced hematological toxicity, which has rarely been reported in transplant recipients, may result in anemia episodes, reported mainly in kidney and heart transplant recipients, sporadic cases of thrombotic thrombocytopenic purpura/hemolytic uremic syndrome, red cell aplasia (4 reported cases), and generalized bone marrow suppression (only 1 reported case). We describe a case of a liver transplant recipient with pancytopenia that appeared during immunosuppressive therapy with tacrolimus. This patient suffered from progressive anemia, leukopenia with severe neutropenia, and mild thrombocytopenia; bone marrow biopsy showed hypoplasia (20% of cellularity) without dysplasia. Bone marrow recovery was made possible by suspending tacrolimus and changing to immunosuppression with cyclosporine A, despite the two drugs being very similar in their mechanism of immunosuppression. Contrary to previously reported cases (pure red cell aplasia and bone marrow hypoplasia), the recovery of hemoglobin and neutrophil values was slow after tacrolimus suspension, even though in the first month transfusions were no longer necessary.     

Bone marrow cell transplantation in clinical perspective

The deficit in left ventricular performance in an ischemically damaged heart is characterized by depletion of functioning cardiomyocytes. The problem is accentuated by the inadequate intrinsic repair mechanism of the heart. Recent progress in regenerative medicine has paved way for an outside intervention to support the limited on-going reparative process in the heart through transplantation of cells with myogenic and/or angiogenic potential. The functional plasticity of bone marrow derived stem cells has been exploited for cardiac repair. The early experiences in proof-of-concept animal studies and phase-1 clinical trials have been encouraging and suggest the safety, feasibility and potential of this approach. We critically review the literature in depth to elucidate the progress in this field together with discussion of the problems and controversies that need to be addressed in order to fully exploit the potential of bone marrow stem cell transplantation with clinical relevance.     

Bone marrow cells produce nerve growth factor and promote angiogenesis around transplanted islets

AIM:To clarify the mechanism by which bone marrow cells promote angiogenesis around transplanted islets.METHODS: Streptozotocin induced diabetic BALB/ c mice were transplanted syngeneically under the kidney capsule with the following: (1) 200 islets (islet group: n=12), (2) 1-5×106 bone marrow cells (bone marrow group: n=11), (3) 200 islets and 1-5×106 bone marrow cells (islet + bone marrow group: n= 13), or (4) no cells (sham group:n=5). All mice were evaluated for blood glucose, serum insulin, serum nerve...     

Bone marrow transplantation or chemotherapy as post-remission treatment of adult acute myelogenous leukemia

Summary We compared three consolidation regimens in patients with acute myelogenous leukemia in first remission. Thirty-four patients received only intensive consolidation chemotherapy (SIC); 28 patients were scheduled to undergo an autologous bone marrow transplant (auto-BMT) and 44 patients an allogeneic BMT (allo-BMT). Twenty-seven of them were referred in first remission for allo-BMT. Nineteen patients achieved a complete remission after salvage treatment. All other patients obtained a remission after one or two courses of a standard combination of cytosine arabinoside and daunorubicin. Except for the patients who were referred in remission, all patients received intermediate dose cytosine arabinoside and amsacrine as a first consolidation treatment. The median ages of the three groups were 48 (SIC), 39 (auto-BMT) and 33 years (allo-BMT). Two patients relapsed before auto-BMT and 1 before allo-BMT. The median interval from the date of complete remission to the auto- or allo-BMT was 3 months. In total, 80% of the patients of the SIC group relapsed, compared to 50% of the patients belonging to the auto-BMT group and 35% of the 44 patients who were scheduled to receive an allo-BMT. The overall median disease-free survival was 14 months, 30% of the patients being alive and disease-free at 3 years. The disease-free survival rate at three years was 25% for the SIC group, 30% for the allo-BMT group and 40% for the ABMT group (P=0.45). Our study shows no benefit for bone marrow transplantation over intensive consolidation treatment. However, large randomized trials are required to define the real value of these treatment modalities.     

Restoration of gut motility in Kit-deficient mice by bone marrow transplantation

Purpose  Interstitial cells of Cajal (ICC) play important roles in autonomic gut motility as electrical pacemakers and mediators of neural regulation of smooth muscle functions. Insufficiency of ICC has been reported in a wide range of gut dysmotilities. Thus, restoration of ICC may be a therapeutic modality in these diseases. Here we provide evidence that transplanted bone marrow (BM) cells can restore gut dysmotility in part via transdifferentiation to ICC. Methods  Bone marrow cells obtained from Kit insufficient W/W ^v mice or syngeneic GFP-transgenic mice with wild-type Kit were transferred to W/W ^v recipients. Whole gut transit time and gastric emptying were examined 5 and 6 weeks after BM transplantation, respectively, and ICCs were identified in whole mounts, frozen sections and transmission electron immunomicroscopy of the gut smooth muscle layers using specific antibodies. Results  Transplantation of wild-type BM into W/W ^v mice significantly improved whole gut transit time and gastric emptying. Fluorescent immunohistochemistry revealed GFP⁺Kit⁺ cells in the myenteric plexus, deep muscular plexus, and submucosal plexus smooth muscle layers of the stomach, small intestine, and colon, respectively. In the whole mounts, GFP⁺Kit⁺ cells were bipolar and spindle shaped, and transmission electron immunomicroscopy showed GFP⁺ cells rich in mitochondria and endoplasmic reticulum between gut smooth muscle layers, suggesting the presence of GFP⁺ cells with morphological characteristics of ICC. Conclusions  These results suggest that BM contains cells that may incorporate into ICC networks and improve dysmotility in W/W ^v mice. Thus, BM transplantation may become to a new therapeutic modality for gut dysmotilities due to ICC insufficiency.     

骨髓干细胞介导的人肝细胞生长因子基因转染在兔心肌梗死组织中的表达

目的:观察以骨髓干细胞(BMSCs)介导的人肝细胞生长因子(hHGF)基因转染在兔心肌梗死组织中能否稳定表达.方法:家兔16只,复制急性心肌梗死(AMI)模型.随机分为2组:移植组:BMSCs加hHGF移植,对照组.另外随机选取8只兔作为假手术组.AMI术后3 d,抽取股骨骨髓1.5 ml,分离BMSCs培养.于对数生长期加入5-溴脱氧尿苷孵育24 h,以标记移植细胞.构建pcDNA3.1-hHGF重组表达载体,移植前用脂质体包裹表达载体转染BMSCs,孵育24 h.AMI术后14 d,行自体细胞移植,对照组接受等量的无血清培养基注射,假手术组只开胸,不注射.细胞移植后28 d,检测左心室血流动力学指标;取心肌组织,RT-PCR检测hHGFmRNA的表达;酶联免疫吸附法(ELISA)测定心肌组织中HGF的含量.结果:与假手术组比较,心肌梗死后28d,对照组左心室功能降低,但BMSCs加hHGF组左心心室功能有改善;RT-PCR结果显示移植组有hHGFmRNA表达,ELISA测定HGF含量移植组高于对照组.结论:AMI组织中HGF含量降低,转染HGF的BMSCs能在AMI组织中生存并能表达分泌HGF,其左心室功能有改善.     

Role of vascular endothelial growth factor (VEGF) and placenta-derived growth factor (PlGF) in regulating human haemopoietic cell growth

Vascular endothelial growth factor (VEGF) and placental derived growth factor (PlGF) stimulate cell proliferation and differentiation by binding to their specific receptors, Flk-1/KDR and Flt-1 respectively. Flk-1/KDR-deficient murine embryos manifest failure of blood-island formation and vasculogenesis. The aim of this study was to directly evaluate the importance of VEGF, PlGF/Flt-1 and Flk-1/KDR receptor ligand interactions in regulating normal and malignant human haemopoiesis. Addition of VEGF and PlGF failed to enhance survival or cloning efficiency of human haemopoietic progenitors isolated from adult bone marrows, fetal livers or cord blood samples. This finding may be explained by the apparent absence of mRNA encoding Flt-1 and Flk-1/KDR receptors on stem cell rich CD34⁺ c-kit-R⁺ Rh123^low cells. Further studies revealed that Flt-1 R mRNA, but not Flk-1/KDR mRNA was first detectable in the more mature cells isolated from haemopoietic colonies. Accordingly, VEGF receptors are either absent, or expressed at very low level, on human haemopoietic stem/progenitor cells. Of interest, normal and malignant human haemopoietic cells appeared to secrete VEGF protein. However, in contrast to normal haemopoietic progenitors, VEGF co-stimulated HEL cell proliferation as well as CFU-GM colony formation from ∼15% of the chronic myeloid leukaemia (CML) and acute myeloid leukaemia (AML) patients studied. Therefore, although VEGF appeared to have minimal effects on normal haemopoietic cell growth it would appear to drive malignant haemopoietic cell proliferation to some degree. Of more importance, however, we speculate that VEGF may play an very important role in leukaemogenesis by stimulating growth of vascular endothelium, thereby providing a sufficient blood supply to feed the growing haematological tumour.     

47例骨髓转移癌的临床及血液学特点

目的 研究骨髓转移癌的临床及血液学特点。方法 骨髓活检塑料包埋切片，H-Gmiesa-E,Gomori,PAS,Alcin blue染色。结果 47例患者中24例找到原发灶，其中胃癌15例，前列腺癌4例，肺癌2例，乳腺癌，结肠癌，肝癌各1例，34例做骨髓活检者骨髓均找到转移癌细胞，而其中10例骨髓象未能发现转移癌细胞，临床表现以贫血最常见（91.5%)，其次是血小板减少（64.7%)，骨痛（61.7%)和发热（48.9%)。结论 对于不明显原因的贫血和骨痛患者，骨髓活检对转移癌具有重要诊断价值。     

间充质干细胞联合同种异体骨髓移植治疗狼疮模型鼠的疗效观察

目的 观察体外扩增的骨髓源间充质干细胞联合同种异体骨髓移植治疗狼疮模型鼠的疗效,并探讨其在系统性红斑狼疮发病机制中的作用.方法 将20只鼠龄为12周的雌性狼疮模型鼠按随机数字表法随机分为单纯骨髓移植组、联合Ⅰ组、联合Ⅱ组、阳性对照组,并设雄性BALB/c小鼠为阴性对照组.将体外扩增的BALB/c小鼠骨髓源问充质干细胞联合雄性C57BL/6小鼠骨髓植入狼疮模型鼠体内,测移植1个月后小鼠体内的Y染色体,并通过小鼠的体重、白细胞、尿蛋白、抗双链DNA抗体水平及终期小鼠肾脏病理和免疫荧光来评价移植的疗效.结果 (1)移植30 d后,可在狼疮模型鼠体内检测到Y染色体,表明移植成功.(2)单纯骨髓移植组、联合Ⅰ组、联合Ⅱ组小鼠尿蛋白和抗双链DNA抗体水平均显著下降.移植40 d后联合Ⅰ组、联合Ⅱ组抗双链DNA抗体吸光度(A)值分别为0.76±0.28、0.73±0.10,与阴性对照组(0.47±0.10)相比,差异无统计学意义(P>0.05);单纯骨髓移植组抗双链DNA抗体水平虽有明显下降(0.91±0.27),但仍高于阴性对照组,差异有统计学意义(P<0.05).移植50 d后单纯骨髓移植组抗双链DNA抗体A值为0.55±0.15,与阴性对照组相比差异无统计学意义(P>0.05).(3)各组接受移植的小鼠肾脏病理光镜下基本恢复正常,未见炎性细胞浸润及血管坏死.免疫荧光强度均弱于阳性对照组.结论 单纯骨髓移植和联合问充质干细胞移植均能有效改善狼疮模型鼠的病情;间充质干细胞可加速狼疮模型鼠外周血抗双链DNA抗体和肾脏免疫复合物的清除,帮助受损器官的恢复.     

Treatment of autoimmune diseases in mice by a new method for allogeneic bone marrow transplantation

Remarkable advances have been made in bone marrow transplantation (BMT), which has become a powerful strategy for the treatment of leukemia, aplastic anemia, congenital immunodeficiency, and also autoimmune disease. Using various animal models, allogeneic (allo) BMT has been found to be useful in the treatment of various autoimmune diseases. In MRL/lpr mice, which are radiosensitive (<8.5 Gy) and are an animal model for autoimmune diseases, conventional BMT resulted in only transient effects; the manifestations of the autoimmune diseases recurred 3 months after BMT. Using MRL/lpr mice, we have very recently established a new strategy for allo BMT. We injected bone marrow cells (BMC) directly into the bone marrow cavity (intrabone marrow [IBM] injection) of recipients that had received fractionated irradiation. This 'IBM-BMT' was found to be effective in treating autoimmune diseases in radiation-sensitive and chimeric-resistant MRL/lpr mice. In addition, this strategy was found to be applicable for the transplantation of organs. We believe that these strategies for BMT and organ transplantation herald a new era in transplantation.     

慢性粒-单细胞白血病的骨髓组织病理学研究

目的:探讨慢性粒-单细胞白血病的(CMML)骨髓组织病理学改变。方法:骨髓活检块以Hema-pun865塑料包埋剂包埋,制备成3μm做HGF染色,5μm做Gomori染色。观察切片内骨髓组织学和细胞形态学改变。并与20例正常人骨髓切片进行对照。结果:46例CMML患者骨髓切片表现增生异常活跃骨髓象,原始细胞增多,ALIP易见,各例均检出典型原始幼稚前体细胞异常定位大簇(>5个髓系前体细胞)位于小梁旁区或间区。粒系细胞和单核系细胞增多(平均值为19.7%)。巨核细胞增多与否不定,但多形性明显。红系呈小簇或散在分布。间质示11例骨小梁破坏,14例静脉窦扩张。网状纤维异常增生(16例,达34.7%,Gomori染色 )。结论:骨髓切片能全面反映CMML骨髓组织病理学改变,提高诊断率,并能提示预后。     

Tolerance and chimerism and allogeneic bone marrow/stem cell transplantation in liver transplantation

The liver has particular tolerogenic properties that allow its spontaneous acceptance in some animal species.Liver structure is considered to favor a tolerogenic environment.The peripheral tolerance mechanisms also play a role in spontaneous tolerance to liver graft.In a clinical setting,the main challenge nowadays facing liver transplantation is minimization of immunosuppression with the goal of donor-specific tolerance.Mechanisms involved in tolerance to transplanted organs are complex and partly unknown.A significant mechanism in tolerance induction is chimerism.Chimerism can be induced through transplantation of allogeneic donor bone marrow/stem cells under appropriate host conditioning.This review focuses on the tolerance mechanisms in liver transplantation and highlights the role of chimerism and allogeneic bone marrow/stem cell transplantation in tolerance development.     

PKH26荧光示踪剂在小鼠骨髓单个核细胞肝内迁移过程中标记作用的研究

目的 探讨PKH26荧光示踪剂在小鼠骨髓单个核细胞肝内迁移过程中的标记作用。方法以红色荧光染料PKH26标记从小鼠骨髓中分离出的骨髓单个核细胞，从小鼠的尾静脉注入CCIA—AAF造成肝损伤的同种异体的小鼠体内，移植2周后取肝组织，通过荧光显微镜观察实验组小鼠骨髓干细胞向肝脏迁移的情况。结果受体组小鼠的肝小叶中央静脉及汇管区均可见新生的PKH26标记阳性的肝细胞。结论PKH26可用于标记向急性肝损伤小鼠肝脏迁移的骨髓单个核细胞。     

Outcome of bone marrow transplantation in patients with extramedullary involvement of acute leukemia

Summary Infiltration of extrahemopoietic tissue with leukemic cells was evaluated as a prognostic indicator in 18 patients with acute leukemia undergoing bone marrow transplantation. When compared to 107 patients who did not have extramedullary leukemia at any time prior to marrow grafting, the patients with leukemic invasion into organs outside the hemopoietic system had a significant increase of leukemic recurrence and a significant decrease in survival after marrow transplantation. Extramedullary leukemia may be a negative prognostic indicator for bone marrow transplantation candidates.This work was supported by NCI Grants CA 30206 and CA 33572     

Bone marrow features of diagnostic impact in erythrocytosis

Controversy continues to persist about the role of histopathology regarding diagnosis of polycythemia vera (PV). For this reason, a clinicopathological study was performed on 334 patients presenting with a sustained borderline to marked erythrocytosis (hemoglobin >17 g/dl in men and >15 g/dl in women). The aim was to elucidate the discriminating impact of bone marrow biopsy examinations in an independent fashion from laboratory parameters. According to morphological findings based on a semiquantitative evaluation of standardized features, cellularity, megakaryocytes (quantity, size, pleomorphous aspect, clustering, nuclear lobulation), eosinophils, cellular debris, perivascular plasmacytosis and iron-laden macrophages exerted a distinctive value. Comparison with clinical data and follow-up revealed that in only 13 patients (4%), histopathology failed to differentiate clearly between PV (208 patients) and secondary polycythemias (113 patients). In conclusion, certain sets of morphological parameters allow a distinction between autonomous and reactive polycythemias and therefore enhance significantly diagnostic validity.     

Bone marrow microvessel density is a prognostic factor for survival in patients with multiple myeloma

 The importance of neoangiogenesis for the progressive growth and viability of solid tumors is well established. Recently, there has been growing evidence that angiogenesis might also be important in hematological malignancies, but only few data are available. In this report, we have studied the impact of bone marrow microvessel density and survival in patients with multiple myeloma (MM). Immunohistochemical CD34 stained paraffin-embedded bone marrow biopsies of 44 patients with newly diagnosed MM were studied. Microvessels were counted in 400× magnification and the mean number of vessels per area in each sample was noted as the microvessel density (MVD). The median MVD was 48 vessels/mm², the range was 0–125 vessels/mm². Using a cut-off value of the median MVD in the Kaplan-Meier analysis, the median survival was 22.2 months in the group with the higher MVD and was not reached in the group with the lower MVD (P<0.01). In a multivariate Cox regression analysis, using previously identified prognostic factors β2-microglobulin, C-reactive protein (CRP), and age, MVD remained significant as a prognostic factor (P<0.03). Received: 3 May 2000 / Accepted: 24 July 2000