This study was undertaken to establish a murine model for unrelated allogeneic umbilical cord blood transplantation (UCBT). The characteristics and percentage of hematopoietic stem/progenitor cells between near-term fetal and neonatal murine peripheral blood (FNPB) and bone marrow (BM) were evaluated by flow cytometry and semisolid methylcellulose culture. BABL/c (H-2(d)) recipient mice conditioned with high dose CTX were transplanted with FNPB form C57BL/6 (H-2(b)) mice and the survival rate, hematopoietic and immunological reconstruction, graft versus host disease (GVHD) and engraftment level were observed. The results showed that the numbers of day 14 CFU-GM and CFU-GEMM in FNPB (176.40 +/- 78.39)% and (141.40 +/- 56.57)%, respectively were much higher than those in BM (75.20 +/- 26.41)% and (68.80 +/- 23.95)%, respectively. Moreover the percentage of Sca-1(+) CD34(+) cell subsets in FNPB (3.63 +/- 1.13)% was also higher than that in BM (1.41 +/- 0.8 7)%. FNPB transplantation improved survival rate and reconstituted hematopoietic and immune function in recipients. There was no evidence of GVHD. Chimeric analysis showed that the proportion of donor cells in BM of recipients was 27.94% at 21 days after transplantation. It was concluded that FNPB contains more hematopoietic stem and progenitor cells with high expansion ability and weak allogeneic immunity, which was similar to human UCB. The murine model for allogeneic UCBT (C57BL/6-->BALB/c) was established successfully.

The purpose of this study is to explore the clinical significance of RBC blood group serological test in nonmyeloablative allogeneic peripheral blood stem cell transplantation (NAPBSCT) of ABO group incompatibility in 4 patients with acute leukemia. ABO and MN blood group of donors and recipients were determined by hemagglutination test and Rh blood group by Diana Gel phenotype Rh card. The changes of blood group in recipients were observed and implant of donor cells was monitored by short tandem repeat-PCR method. The results showed that in 2 cases of 4 recipients the marrow cells appeared mixed chimera of donor and recipient cells, and blood group changed to donor type in 1 of the 2 cases on 100 days after transplantation. In another 2 cases, the marrow cells appeared mixed chimera without blood group chimera on 154 days after transplantation, and rejection of the transplant occurred in 1 of the 2 cases. The determination of hemagglutinin titer showed that the implant rate of donor cells was lower in the recipients with higher hemagglutinin titer and blood group chimera did not appear, conversely, the implant rate was higher in the recipients with lower titer and blood group chimera appeared early. It is concluded that examination of RBC blood group in NAPB SCT can indirectly reflect effectiveness of transplantation, contribute to decide the intensity of conditioning protocol and immunosuppressive therapy after transplantation, estimate prognosis and guide blood transfusion during transplantation.

In order to understand the effect of hematopoietic stem cell mobilization agents, G-CSF and GM-CSF, on the expression of TNF-alpha mRAN and CD69 and secretion of IgG in SLE patients' peripheral blood mononuclear cells (PBMNC), expression of TNF-alpha mRNA and CD69 was measured by RT-PCR and flow cytometry, respectively, and IgG secretion by ELISA. The results showed that 0.1 - 2.0 microg/ml G-CSF did not affect the expression of TNF-alpha mRNA in active and static patients' PBMNC treated with 0.1 - 2.0 microg/ml cytokines, and 2.0 microg/ml GM-CSF increased the expression of TNF-alpha mRNA in active patients' PBMNC. G-CSF and GM-CSF did not interfere the expression of CD69 in active and static patients' PBMNC, however, the expression of CD69 was significantly increased in active patients' PBMNC treated with GM-CSF at more than 8 microg/ml. There was no obvious change of IgG secretion from PBMNC induced with 10 microg/ml G-CSF, while the IgG secretion was stimulated by 10 microg/ml GM-CSF. It was concluded that G-CSF as a mobilization agent could be safe to SLE patients, but GM-CSF may cause some harmful effects to patients because of the increase of the parameters relating to activity of lupus in active SLE patients.

To compare the expression of CD antigens on immune cells from umbilical cord blood (UCB) and bone marrow (BM) and analyze its clinical significance, the phenotypes of lymphoid cells and nucleated cells from 38 UCB and 10 BM samples were investigated by flow cytometry with double labeling monoclonal antibodies. The results showed that the immature lymphocytes (CD3(-) CD4(+)) were detected in UCB and higher than those in BM; cytotoxic T lymphocytes (CD3(+) CD16(+) CD56(+)) in UCB were significantly lower than those in BM. NK cells (CD3(-) CD16(+) CD56(+)) in UCB were higher than those in BM. The ratio of CD34(+) cells in nucleated cells of UCB was similar to that of BM, however, both the contents of myeloid (CD34(+) CD13(+) and CD34(+) HLA-DR(+)) and lymphoid (CD34(+) CD19(+)) progenitor cells in UCB were lower than those in BM. It is concluded that the immune cells in UCB possess immaturity, which might lead to mild GVHD after UCB transplantation. It is inferred from the higher ratio of NK cells in UCB, GVL will not decrease after UCB transplantation. The lower contents of myeloid and lymphoid progenitor cells in UCB probably accounted for the slow hematopoiesis and immune reconstitution following UCB transplantation.

To explore the effect of retinoic acid (RA) on the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in murine bone marrow stromal cell (BMSC) and adhesive rate of human cord blood mononuclear cell (UCBMNC) to BMSC in vitro, the express ions of ICAM-1 and VCAM-1 of murine BMSC were detected by flow cytometry and the binding capacity of UCBMNC to BMSC was tested by MTT assay after co-culturing with 0.1, 1.0 and 10.0 micro mol/L RA, respectively. The results showed that 1.0 and 10.0 micro mol/L RA increased the expression of ICAM-1 and the adhesive rate of U CBMNC to BMSC, however, RA did not induced the increase of expression of VCAM-1. It was positive correlation between the increments of ICAM-1 expression and the adhesive rate (r = 0.7883, P < 0.05). It is concluded that RA up-regulated ICAM-1 expression of BMSC and increased the adhesion of UCBMNC to BMSC in vitro. These may clarify the correlation between adhesion molecules on BMSC and homing of hematopoietic stem cells, and provide the experimental basis for RA to promote the homing of umbilical cord blood stem cell.

To evaluated the feasibility of preventing infection after high dose chemotherapy and radiotherapy using the granulocytes derived from differentiated from hematopoietic stem/progenitor cells ex vivo, human CD34-positive cells were isolated from umbilical cord blood by using a high-gradient magnetic cell sorting system (MACS), and the cells committedly differentiated with hematopoietic cytokines (SCF + IL-3 + IL-6 + G-CSF) in a liquid culture system. The expanded cell number, ratio of the viable cells, chromosome and phenotype of the differentiated cells and safety analysis of expanded cells were detected by using cell count, trypan blue exclusion test, karyotype analysis, flow cytometry and tumorigenic model of nude mice, respectively. The results showed that the combination of cytokines increased cell number by (1006.4 +/- 103.2) folds and flow cytometric analysis showed myeloid marker CD11b expressed in the about 60% cells. The growth peak of differentiated cells was at 14 days of culture and decreased at about 33 days. No abnormality was found in the karyotype analysis of expanded cells. No tumor was found in the nude mice injected with expanded cells after 35 days and the expanded cells had the ability of phagocytizing bacteria. It is concluded that the cells, differentiated from CD34(+) cells, expanded ex vivo possess the function of granulocyte and it was safe for clinical trial.

In this study, the roles of hematopoietic inhibitors elaborated by bone marrow endothelial cells in the proliferation and differentiation of hematopoietic progenitors were investigated. Murine bone marrow endothelial cell conditioned medium (BMEC-CM) was collected and the components with > 10 kD and < 10 kD were obtained by centrifugal ultrafiltration. The effect of BMEC-CM and its components on proliferation of hematopoietic progenitors was evaluated by CFU-GM and HPP-CFC assay and antibody neutralization test. The expression of the inhibitors in BMEC and BMEC-CM was detected by RT-PCR and Western blot, and change of proliferation and differentiation-related genes during expansion of hematopoietic progenitors was examined by membrane hybridization technique.

To study the possibility that induced marrow stromal cells (MSCs) to neuron in vitro by baicalin, a kind of flavonoid isolated from an important medicinal plant Scutellariae Radix.

To investigate the effect of transfection of 5-aza cytidine (5-aza) induced BMSCs that were transfected with Ad. ANG ex vivo into infarcted myocardium.

To establish embryonic stem (ES) cell lines from human fertilized ovum in China.

To study the feasibility and clinical outcome of granulocyte-colony stimulating factor (G-CSF) mobilized allogeneic bone marrow (G-BM) plus G-CSF mobilized peripheral blood stem cells (G-PBSC) transplantation for malignant hematological disease.

To observe the safety of inoculation of recombinant adenovirus with human neurotrophin-3 (Ad-NT3) into cochlea and its effect on hearing function.

To investigate the effects of bone morphogenetic protein (BMP) on the proliferation and collagen synthesis of skeletal muscle satellite cells.

Protein kinase C (PKC) is a serine/threonine kinase family consisting at least 11 related isoenzymes, and it can be divided into four distinct classes, conventional PKC (cPKCs), novel or new PKC (nPKCs), atypical PKC (aPKCs), and PKC-u. All PKC members contain conserved and variant amino acid sequences in ATP binding sites, phosporyl transfer region, pseudosubstrate region, and phorbol ester binding sites. PKC isoenzymes exhibit distinct tissue distribution and play a critical role in cellular events. This review mainly summarized the PKC effects on tumorigenesis, tumor invasion and metastasis, tumor multidrug resistance, differentiation and development of hematopoietic progenitor cells, and hormonal production and secretion.

To assess the culture and differentiation of neural stem cells in embryonic mice and set up a basis for further research in to neural stem cells.

To understand and improve the transdifferentiation of pancreatic stem cells into islet cells through isolation, cultivation and transdifferentiation of adult human pancreatic duct cells.

To explore the morphological, immunophenotypical and cytogenetic (MIC) characteristics of chronic myeloid leukemia in blast crisis (CML-BC).

To detect the contaminating minimal residual disease (MRD) in autologous peripheral blood stem cells (APBSC) and evaluate its impact on the prognosis of non-Hodgkin's lymphoma NHL patients.

The influence of the mimic hematopoietic microenvironment and adhesion factor on the initial divisional behavior of human cord blood hematopoietic progenitors was studied in the culture system with certain cytokines.

To analyze the expression of the CD(40) and its ligand (CD(40)L) on bone marrow stromal cells (BMSC), and investigate interleukin-3 (IL-3), IL-6, Flt3 ligand (FL) and stem cell factor (SCF) production of BMSC after stimulated with CD(40) agonistic monoclonal antibody (5C11) and its role in the regulation of hematopoiesis.