Humans ; Fetal Blood ; Hematopoietic Stem Cell Transplantation ; Transplantation, Homologous ; Anemia ; Cord Blood Stem Cell Transplantation/methods* ; Graft vs Host Disease

OBJECTIVETo explore the effects of umbilical cord blood mononuclear cells (UCBMC) transplantation on the neuronal apoptosis and the expression of Bcl-2 and Bax proteins in neonatal rats with hypoxic-ischemic brain damage (HIBD).

METHODSSeven-day-old Sprague-Dawley neonatal rats were randomly divided into normal control (N)+normal saline (NS), HIBD+NS, N+UCBMC, and HIBD+UCBMC groups. HIBD model was prepared using the classical Rice-Vannucci method. Twenty-four hours after HIBD, UCBMC were transplanted in the N+UCBMC and HIBD+UCBMC groups. Seven days after transplantation, NeuN/Cleaved-Caspase-3 double immunofluorescence staining and TUNEL methods were used to observe neural apoptosis in the cortex. The expression levels of Bax and Bcl-2 proteins were examined by Western blot analysis.

RESULTSThere were more NeuNcleaved Caspase-3DAPIand TUNELDAPIcells in the HIBD+NS group compared with the N+NS and N+UCBMC groups (P<0.01). There were less NeuNcleaved Caspase-3DAPIand TUNELDAPIcells in the HIBD+UCBMC group compared with the HIBD+NS group (P<0.01). The concentration of Bax protein was higher and that of Bcl-2 proteins was lower in the HIBD+NS group compared with the N+NS and N+UCBMC groups (P<0.01). The concentration of Bax protein in HIBD+UCBMC group was lower than that in the HIBD+NS group (P<0.01). The concentration of Bcl-2 protein was higher compared with the HIBD+NS, N+NS and N+UCBMC groups (P<0.05).

CONCLUSIONSUCBMC transplantation via lateral ventricle can upregulate the expression of Bcl-2 protein and down-regulate the expression of Bax protein, thus alleviating brain neural apoptosis in neonatal rats with HIBD.

Animals ; Animals, Newborn ; Apoptosis ; Caspase 3 ; metabolism ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; therapy ; Male ; Neurons ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; analysis

Although mesenchymal stem cells (MSCs) are increasingly used to treat graft-versus-host disease (GVHD), their immune regulatory mechanism in the process is elusive. The present study aimed to investigate the curative effect of third-party umbilical cord blood-derived human MSCs (UCB-hMSCs) on GVHD patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and their immune regulatory mechanism. Twenty-four refractory GVHD patients after allo-HSCT were treated with UCB-hMSCs. Immune cells including T lymphocyte subsets, NK cells, Treg cells and dendritic cells (DCs) and cytokines including interleukin-17 (IL-17) and tumor necrosis factor-alpha (TNF-α) were monitored before and after MSCs transfusion. The results showed that the symptoms of GVHD were alleviated significantly without increased relapse of primary disease and transplant-related complications after MSCs transfusion. The number of CD3(+), CD3(+)CD4(+) and CD3(+)CD8(+) cells decreased significantly, and that of NK cells remained unchanged, whereas the number of CD4(+) and CD8(+) Tregs increased and reached a peak at 4 weeks; the number of mature DCs, and the levels of TNF-α and IL-17 decreased and reached a trough at 2 weeks. It was concluded that MSCs ameliorate GVHD and spare GVL effect via immunoregulations.
Adolescent ; Adult ; Cord Blood Stem Cell Transplantation ; methods ; Cytokines ; metabolism ; Dendritic Cells ; metabolism ; Female ; Graft vs Host Disease ; immunology ; therapy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Immunomodulation ; Killer Cells, Natural ; metabolism ; Male ; T-Lymphocyte Subsets ; metabolism ; Transplantation, Homologous ; adverse effects ; Young Adult

Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) may be a promising modality for treating medial temporal lobe epilepsy. 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) is a noninvasive method for monitoring in vivo glucose metabolism. We evaluated the efficacy of hUCB-MSCs transplantation in chronic epileptic rats using FDG-PET. Rats with recurrent seizures were randomly assigned into three groups: the stem cell treatment (SCT) group received hUCB-MSCs transplantation into the right hippocampus, the sham control (ShC) group received same procedure with saline, and the positive control (PC) group consisted of treatment-negative epileptic rats. Normal rats received hUCB-MSCs transplantation acted as the negative control (NC). FDG-PET was performed at pre-treatment baseline and 1- and 8-week posttreatment. Hippocampal volume was evaluated and histological examination was done. In the SCT group, bilateral hippocampi at 8-week after transplantation showed significantly higher glucose metabolism (0.990 +/- 0.032) than the ShC (0.873 +/- 0.087; P < 0.001) and PC groups (0.858 +/- 0.093; P < 0.001). Histological examination resulted that the transplanted hUCB-MSCs survived in the ipsilateral hippocampus and migrated to the contralateral hippocampus but did not differentiate. In spite of successful engraftment, seizure frequency among the groups was not significantly different. Transplanted hUCB-MSCs can engraft and migrate, thereby partially restoring bilateral hippocampal glucose metabolism. The results suggest encouraging effect of hUCB-MSCs on restoring epileptic networks.
Animals ; Chronic Disease ; Cord Blood Stem Cell Transplantation/*methods ; Epilepsy, Temporal Lobe/*metabolism/pathology/*therapy ; Fluorodeoxyglucose F18/*pharmacokinetics ; Hippocampus/*metabolism/*pathology/surgery ; Male ; Mesenchymal Stem Cell Transplantation/methods ; Radiopharmaceuticals/pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Sensitivity and Specificity ; Tissue Distribution ; Treatment Outcome

This study was purposed to explore the effectiveness of mixed transplantation of HLA mismatched bone marrow hematopoietic stem cells(HSC), peripheral blood HSC and umbilical cord blood HSC for treatment of pediatric blood diseases. From August 2012 to December 2012, five children with refractory hematological diseases in our hospital received allogeneic hematopoietic stem cell transplantation. The mixed grafts consisting of HLA-mismatched bone marrow HSC, peripheral blood HSC and umbilical cord blood HSC were used to observe the effects of umbilical cord blood HSC on the time of hematopoietic reconstruction of bone marrow, STR chimeric degrees, incidence of GVHD. and early transplant-associated complications. The results showed that all 5 children patients were grafted successfully with the median grafted time of 11 d for ANC>0.5×10(9)/L and 10 d for Plt>20×10(9)/L, respectively. On day 30, the STR-PCR test of peripheral blood showed a stable complete chimera. Five cases suffered from mild to moderate symptoms of GVHD, showing with I-II grade of skin GVHD and in which two cases suffered from diarrhea, showing I-II grade of intestinal GVHD. All the 5 patients had no liver function damage. One patient died of severe hemorrhagic cystitis and multi-site infection, and the remaining four cases survived so far on the current median follow-up time of 137 d (130 d-250 d). It is concluded that transplantation of the mixed HLA mismatched bone marrow HSC, peripheral blood HSC, with third-party cord blood HSC can increase the survival rate for pediatric patients with blood disease.
Child ; Cord Blood Stem Cell Transplantation ; Female ; Hematologic Diseases ; therapy ; Hematopoietic Stem Cell Transplantation ; methods ; Histocompatibility Testing ; Humans ; Male ; Peripheral Blood Stem Cell Transplantation ; Treatment Outcome

This study was purposed to investigate the clinical value of HLA matching(low and high resolution) and its effect on outcome of the patients received umbilical cord blood transplantation(UCBT). Sequence-specific oligonucleotide probe (SSOP) , sequence-based typing (SBT) and sequence-specific primers(SSP) were used to perform high resolution HLA matching for HLA-A, -B, -Cw, -DRB1, -DQB1 and low resolution for HLA-A, B, DRB1 among 34 patients with hematologic malignancies who received unrelated UCB transplantation and grafts. The effects of HLA matching (low or high resolution ) on leading engraftment, hematopoietic reconstitution, graft-versus-host disease (GVHD) and infection after UCB transplantation were analyzed by comparison. The results showed that the median of total nucleated cells (TNC) of transplanted cord blood was 6.0×10(7)/kg, The time of neutrophil recovery was significantly shortened when more than 5×10(7)/kg TNC were transplanted (P < 0.05). The HLA-(6-10)/10 group of high resolution HLA matching was better than the HLA (4-5)/10 group in the respect of leading engraftment, the time of platelet recovery and the rate of acute GVHD (P < 0.05). In contrast, HLA-I+II locus, HLA-DRB1 or HLA-DQB1 locus mismatch could prolong the platelet engraftment time (P < 0.05). There was statistical difference in the time of platelet recovery, the rate of acute GVHD between the HLA (5-6)/6 group of low resolution HLA matching and the HLA (3-4)/6 group after UCB transplantation (P < 0.05), but the mismatch locus of HLA with low resolution did not correlate with the time of platelet recovery (P > 0.05). It is concluded that the high resolution HLA matching between patients received unrelated UCB transplantation and grafts may contribute to select the better UCB, that has important clinical value to promote hematopoietic reconstitution and to reduce the complications after UCB transplantation.
Adolescent ; Adult ; Child ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Hematologic Neoplasms ; therapy ; Histocompatibility Testing ; methods ; Humans ; Male ; Young Adult

Adolescent ; Adult ; Child ; Cord Blood Stem Cell Transplantation ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia, Myeloid, Accelerated Phase ; therapy ; Male ; Middle Aged ; Siblings ; Tissue Donors ; Transplantation, Homologous ; Young Adult

This study was purposed to investigate the efficacy and safety of haploidentical hematopoietic stem cells (allo-HSCT) transplantation combined with human umbilical cord-derived mesenchymal stem cell infusion (hUC-MSC) for severe aplastic anemia-II (SAA-II). Eight SAA-II patients received haploidentical allo-HSCT, the G-CSF mobilized peripheral hematopoietic stem cells and bone marrow haploidentical hematopoietic stem cells were selected as graft, the human umbilical cord-derived mesenchymal stem cells (hUC-MSC) were infused as the third party. Conditioning regimen consisted of rabbit anti-thymic lymphocytes protein(ATG), cyclophosphamide(CTX) and fludarabine(Flu). For two patients out of 8 SAA-II patients the conditioning regimen was combined with busulfan(BU). The graft versus host disease(GVHD) was prevented with CSA, MTX, ATG, CD25 and mycophenolate mofetil. The results showed that the average number of nucleated cells were 9.13×10(8)/kg, and number of CD34(+)cells were 3.76×10(6)/ kg. All the 8 SAA-II patients achieved hematopoietic reconstitution. The average time of neutrophils count>0.5×10(9)/L was 11.9 days, and average time of Plt level >20×10(9)/L was 14.6 days. The incidence of acute GVHD of I-II grade was 25%, and that of III-IVgrade was 12.5%, the transplantation-related mortality was 25%. It is concluded that haploidentical allo-HSCT combined with umbilical cord MSC infusion is an effective approach to cure SAA.
Adolescent ; Adult ; Anemia, Aplastic ; therapy ; Child ; Cord Blood Stem Cell Transplantation ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Middle Aged ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; Young Adult

This study was purposed to investigate the engraftment, graft-versus-host disease (GVHD), transplantation related mortality (TRM), relapse and survival in hematologic patients received unrelated umbilical cord blood transplantation (UCBT). A total of 25 patients with hematological disease underwent UCBT, including 8 pediatric and 17 young adult patients. Among them 3 cases received single unit of UCBT and 22 cases received double units of UCBT. For donor/recipients human leukocyte antigen (HLA) matching: HLA 6/6 loci matched in 9 cases, HLA 4-5/6 loci matched in 16 cases. There were 19 patients with hematologic malignancies, including 3 cases in the period of disease progression and 6 cases of non-hematologic malignancies. Conditioning regimens were TBI/Cy ± Flu ± ATG or BuCy ± Flu ± ATG for 21 patients and Cy+Flu+ATG for 4 patients. For prophylaxis of acute graft-versus-host disease (aGVHD) the regimen of cyclosporine (CsA) as dominant drug was used. The results showed that among 16 patients (80.0%) achieved engraftment, 20 patients survived for more than 42 d after transplantation. The cumulative neutrophil recovery rate on day 42 after transplant was 64.0%, with a median time of 17.0 d;the cumulative platelet recovery rate on day 100 after transplant was 60.0 %, with a median time of 35.0 d. The cumulative rate of grade II-IV and III-IV aGVHD after transplantation 100 d was 44.0% and 30.7%, respectively. Until the end of the follow-up, the cumulative rate of TRM was 54.3%. For all the patients, overall survival rate was 42.7%. Out of 17 evaluable patients with hematologic malignancies 7 cases (41.2%) survived to date, and only 1 case relapsed, so event-free survival rate was 35.3%. Out of 5 evaluable patients with non-hematologic malignancies, 4 patients survived and 2 patients were in stable engraftment state, 2 cases with autologous hematopoietic recovery. Among 3 cases of hematologic malignancies at advanced stage, only 1 case survived to date. It is concluded that HLA-4-6/6 loci matched UCBT is an effective option to treat hematological diseases. Double cord blood transplantation (dUCBT) can overcome the disadvantage of insufficient cells of single cord blood UCBT to treat overweight children and adult.
Adolescent ; Adult ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Fetal Blood ; Graft vs Host Disease ; Hematologic Diseases ; therapy ; Histocompatibility Testing ; Humans ; Male ; Survival Rate ; Young Adult

OBJECTIVETo investigate effect of Compound Danshen Dripping Pill (CDDP) on the inflammatory response of the myocardium of acute myocardial infarction (AMI) rabbits, to observe the therapeutic effect of CDDP combined intravenous transplantation of human umbilical cord blood mononuclear cells (HUCBMCs) on inflammatory response, pro-inflammatory cytokine tumor necrosis factor alpha (TNF-alpha) , and heart function in the myocardium of AMI rabbits, and to explore the possible protective mechanisms of the combined therapy.

METHODSThe AMI model was successfully established by ligation of the left anterior coronary artery (LAD) in 40 healthy rabbits.Then they were randomly divided into four groups, i.e., the control group, the CDDP group, the transplantation group, and the combined group, 10 in each group. Rabbits in the control group received intravenous injection of 0.5 mL normal saline via ear vein within 24 h after AMI and then intragastric infusion of normal saline at 5 mL per day. Rabbits in the CDDP group received intravenous injection of 0.5 mL normal saline via ear vein within 24 h after AMI and then intragastric infusion of solution obtained by solving 270 mg CDDP in 5 mL normal saline per day. Rabbits in the transplantation group received intravenous injection of 0.5 mL normal saline labeled with green fluorescent protein (GFP) containing 3 x 10(7) of HUCBMCs via ear vein within 24 h after AMI and then intragastric infusion of normal saline at 5 mL per day. Rabbits in the combined group received intravenous injection of 0.5 mL normal saline labeled with GFP containing 3 x 10(7) of HUCBMCs via ear vein within 24 h after AMI and then intragastric infusion of solution obtained by solving 270 mg CDDP in 5 mL normal saline per day. At week 1 and 4 after treatment, cardiac function indices such as left ventricular fractional shorting (LVFS) and left ventricular ejection fraction (LVEF) were performed by echocardiography; the number of transplanted cells in the myocardium was found by GFP positive cells counted with fluorescence microscopy.The white blood cells in the myocardium stained with HE were determined by light microscope. The expressions of TNF-alpha protein in the myocardium were detected by immunohistochemical assay.

RESULTS(1) Compared with the control group at week 1 and 4 after treatment, the LVEF and LVFS were significantly improved in the CDDP, transplantation, and combined groups (P < 0.05). The cardiac function was significantly improved in the combined group than in the CDDP group and the transplantation group (P < 0.05). But there was no statistical difference in the latter two groups. (2) Compared with the control group, the number of white blood cells and the expression of TNF-alpha protein decreased significantly in the CDDP, transplantation, and combined groups at week 1 and 4 respectively after treatment. The number of white blood cells and expressions of TNF-alpha protein were significantly lower in the combined group than in the CDDP group and the transplantation group (P <0.05). But there was no statistical difference in the latter two groups. (3) GFP-positive cells were found to be distributed in the peri-myocardial infarction area in the transplantation group and the combined group at week 1 and 4 after transplantation. Besides, the number of the GFP positive cells was much more in the combined group than in the transplantation group (P < 0.05).

CONCLUSIONSThe findings indicated that the combination of CDDP with intravenous transplantation of HUCBMCs in the treatment of AMI rabbits could elevate the survival rate of transplanted cells, and further improve the heart function. The possible mechanisms might be related to attenuating local inflammation of myocardium, and inhibiting enhanced expressions of pro-inflammatory cytokine TNF-alpha protein.

Animals ; Cord Blood Stem Cell Transplantation ; methods ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Inflammation ; Myocardial Infarction ; pathology ; therapy ; Rabbits ; Salvia miltiorrhiza ; chemistry ; Transplantation, Heterologous ; Tumor Necrosis Factor-alpha ; metabolism