To explore the expression profile of microRNAs during the course of embryonic stem cells differentiation towards hepatocytes induced by sodium butyrate.

To study the effect of rapamycin in inducing naïve murine effector T cell (Teff) convert to regulatory T cell (Treg) in vitro.

To investigate the effect of murine mesenchymal stem cells (mMSC) on immunoproliferative response of spleen cells.

To explorer the effectiveness of enriched bone marrow stem cells technique for lumbar fusion.

URPOSE: To isolate and culture dental pulp stem cells from human deciduous teeth, and induce them to osteoblasts.

To culture bone marrow mesenchymal stem cells (BMSCs) of rat in vitro and observe HSV-1 infection on BMSCs, BMSCs were separated from the bone marrow and identified by alizarin red staining and detection of ALP. The morphology of HSV-1 infected BMSCs and the CPE were observed. The total DNA was extracted from HSV-1 infected BMSCs and the desired specific gene fragment of 477bp of HSV-1 was amplified by PCR. Results showed that after BMSCs were induced by mineral-fluid for 14 days, the ALP level was increased and the nodule calcification was formed. The induced BMSCs were manifested to have the characteristics of osteoblasts. CPE couldn't be found in HSV-1 latently infected BMSCs but the 477bp gene fragment was still detectable. HSV-1 could establish latent infection in BMSCs after 7 passages. This study indicated that rat BMSCs could be induced to differentiate into osteoblasts in vitro, therefore they can be used as the seed cells for the tissue engineering. HSV-1 can infect rat BMSCs and develop the latent infection in vitro.

To investigate the effects of different nuclear factor (NF)-KB dimers on the survival of immortalized neural progenitor cells (INPCs).

To investigate the impact of transplantation of immortalized neural progenitor cells (INPCs) into the brain with focal cerebral ischemia and the survival and differentiation thereof.

To study the developmental electrophysiological properties of delayed rectifier outward K+ currents (IDR) in undifferentiated NSC and NSC-derived neurons at various time points of adult SD rat hippocampus in vitro.

To investigate the relation between transdifferentiation of the airway myofibroblasts and the expression level of (trypsinogen16, TG16) in vitro and explore the mechanism of airway basement membrane thickening.

To observe the changes of auditory electrophysiology and inner ear pathology in rat cochlea after the noise exposure, and to offer the experimental data for exploring the mechanism of noise-damaged cochlea.

Based on the requirement of culture conditions for hematopoietic stem and progenitor cells (HSPCs) ex vive expansion, we developed a new-type bioreactor by combining superiorities of static and stirred culture models. Stem cell factor (SCF), thrombopoietin (TPO), FLT-3 ligand(Flt-3) were used as the cytokines cocktails. The effects of the static and cyclic culture on the expansion characteristics of CD34+ selected cells were compared in the new-type bioreactor. After 7 d cultures, the expansion of total cells in the static culture was 13.86 +/- 4.26 fold, higher than that in the cyclic culture (7.23 +/- 2.67 fold). The analysis of the fold expansion and the proportion of CD34+ cells showed that there was no marked difference between the static culture and the cyclic culture. However, the fold expansion and the proportion of CD34+CD38- cells were higher in the cyclic culture than those in the static culture (3.90 +/- 0.85 versus 1.82 +/- 0.58), which reflected more primary CD34+CD38- cells were obtained in the cyclic culture. The above results demonstrated that both the static culture and the cyclic culture could be used in ex vive expansion of CD34+ cells with the new-type bioreactor. In static culture hematopoietic stem cells differentiated into progenitor cells, whilst the cyclic culture favored the expansion of primary HSPCs.

To study the anatomy of Dracaena cochinchinensis systematically, and find out the distribution and detect the constituents of its resin, in order to provide substantial foundation for the formation mechanism of its red resin.

Allogeneic haematopoietic stem cell transplantation (allo-HCT) is the most effective curative therapy in myelodysplastic syndromes (MDS). Incidence of MDS increases with age, peaking in the seventh decade of last century. Despite improved consolidation chemotherapy regimens, the prognosis of MDS in patients beyond 60 years of age is dismal. The introduction of peripheral blood-derived stem cell grafts into allogeneic HSCT and the known anti-tumor effect of donor lymphocyte infusions paved the way for reduced-intensity conditioning (RIC) allogeneic hematopoietic stem-cell transplantation, which makes transplant possible in advanced age, significantly alleviates transplant-related organ toxicity and decreases non-relapse mortality. This article reviews the advanced development of reduced-intensity conditioning regimens in allogeneic hematopoietic stem cell transplantation for myelodysplastic syndromes and the future of reduced intensity conditioning hematopoietic stem cell transplants including feasibility of RIC allo-HSCT in treating patients with MDS, selection of MDS cases for RIC allo-HSCT, opportunity of RIC allo-HSCT, source of stem cells for RIC allo-HSCT, RIC regimen for allo-HSCT, evaluation of curative efficacy and prognosis, GVHD and graft versus MDS, and so on.

NOD/SCID (non-obese diabetic/severe combined immunodeficient) mice are immune deficient mice which are made by backcross of severe combined immunodeficient mice with non-obese diabetic mice strains. NOD/SCID mice are both innate immune deficiencies and lack of T and B lymphocytes. Various tumor cells can be implanted in this kind of mice, the rejection and graft-versus-host disease (GVHD) occur fewer. Therefore, NOD/SCID mice gradually become a useful tool for the study on Experimental Hematology. This paper comprehensively reviews the biological characteristics of NOD/SCID mice, the establishment of human leukemia model, stem cell transplantation, drug research, deficiency and improvement of NOD/SCID mice in application for study.

As a member of the hox gene family, hoxB4 gene encodes a class of DNA-dependent homeobox domain nucleoprotein, which is a specific transcription factor, playing an important role in regulating the balance between self-renewal and differentiation of hematopoietic stem cells (HSCs). Therefore, it is important to understand the mechanisms involved in regulating expression of hoxB4 in the HSC. Previous studies have suggested that some hoxB4 upstream regulatory factors, such as USF-1 (upstream activating factor -1), USF-2 (upstream activating factor -2) and NF-Y complex, as well as hematopoietic cytokines, such as platelet growth factor (TPO) and Wnt3a protein, play important regulatory roles in the expression of hoxB4 in hematopoietic stem cells. In this review the structure and biological characteristics of hoxB4, mechanisms involved in regulating expression of hoxB4 in the HSC are summarized.

In order to construct a pichia pastoris expression vector containing the extracellular domain of human Jagged1 and the Fc fragment of human IgG1 fusion gene, or containing only the Fc fragment of human IgG1 and to express them in pichia pastoris. The extracellular domain of human Jagged1 gene was cloned from normal human bone marrow cells. After DNA sequencing, the extracellular domain of Jagged1 gene was inserted into pIC-Fc vector constructed previously, which is Pichia pastoris expression vector containing only the Fc fragment of human IgG1. The constructed plasmid was transformed into yeast GS115 by means of electroporation. The recombinant transformants with a high copy number of the plasmid were selected on MD plate with G418. The expression of protein was induced by addition of methanol. Then, protein expression was analyzed by SDS-PAGE. The results indicated that the extracellular domain of human Jagged1 gene was effectively amplified. The DNA sequencing result showed that the constructed plasmid containing hJagged1(ext)-Fc fusion gene was the same as designed. The fusion protein was successfully expressed in Pichia pastoris. It is concluded that the hJagged1(ext) gene has been successfully cloned and expressed, which provides a new fusion protein for further experiments, the hJagged1(ext)-Fc fusion protein can be used as a new stimulator for proliferation of hematopoietic stem/progenitor cells in vitro.

The objective of this study was to investigate the effect of dendritic cells (DCs) on expansion and function of autologous natural killer (NK) cells and its mechanism in vitro. NK cells were expanded from peripheral blood mononuclear cells (PBMNCs) of healthy volunteers in stem cell growth medium (SCGM) supplemented with rhIL-2 (control group) in 24-well culture plates at 37 degrees C in a humidified CO(2)-containing atmosphere. NK cells were cultured with autologous DCs in the ratio of 5 to 1 (group 5:1) or 1 to 1 (group 1:1) from day 10 after expansion. Total cells of every group were counted and the expression of CD3, CD16/56 on the surface of NK cells was assayed by flow cytometry on days 7, 14 and 21 to calculate the expansion of NK cells. Cytotoxicity of expanded NK cells against K562 cells was assayed by MTT method. TNF-alpha and IL-12p70 were detected in culture supernatants by sandwich ELISA. The results indicated that the expansion and cytotoxicity of NK cells were improved after mixed with autologous DCs. Furthermore, when DCs were mixed with NK cells, the ratio of DCs to NK cells was higher, the expansion and cytotoxicity NK cells were higher. On day 14, the expansion multiple in control, group 5:1 and group 1:1 were 16.26 +/- 1.58, 29.25 +/- 4.01 and 21.23 +/- 2.91 respectively. The expansion multiple of group 5:1 was much higher than that of the other two groups (p < 0.05). The expressions of CD3(-), CD56/16(+) on surface of NK cells in control, group 5:1, group 1:1 were (34.8 +/- 5.1)%, (64.6 +/- 7.8)% and (50.6 +/- 8.7)% respectively and that of group 5:1 was the highest (p < 0.05). The cytotoxicities against K562 cells in control, group 5:1 and group 1:1 were (63.7 +/- 3.8)%, (87.4 +/- 6.8)% and (75.4 +/- 6.3)% respectively. The cytotoxicity of group 5:1 was higher than that in the other two groups also (p < 0.05). TNF-alpha and IL-12p70 levels in culture supernatants when DCs and NK cells were mixed in the ratio of 5 to 1 were much higher than those in culture supernatants of DCs and NK cells alone or in culture supernatants when DCs and NK cells were mixed in the ratio of 1 to 1 (p < 0.05). It is concluded that the expansion and cytotoxicity of NK cells can be improved by DCs and it depended on the mixed ratio of DCs to NK cells. The elevated expansion of NK cells by DCs bears relation to IL-12 produced by DCs. The enhanced cytotoxicity of NK cells is associated with TNF-alpha secreted by NK cells.

The aim of this study was to expand hematopoietic progenitor cells at large scale by magnet stirred culture system. Mononuclear cell from umbilical cord blood were cultured in serum-free medium with stem cell factor, FIT-3 ligand and thrombopoietin. Firstly, the role of magnet on cell growth and colony-forming was studied by static culture on 0, 25 and 50 mT. Then the expansion multiple of cells, colony-forming and expression of surface markers were studied in magnet stirred culture by cell counting, colony-forming assay and flow cytometry. The results indicated that there was no difference in multiple of total cell expansion and numbers of hematopoietic colonies between 0, 25 and 50 mT groups and spinner groups (all p > 0.05). After 7 day cultures, the multiple of total cell expansion in magnet stirred culture was higher than that in static culture (p < 0.01). The numbers of CFU-GM (colony-forming unit-granulocyte/macrophage) and CFU-E (erythroid colony forming unit) in magnet stirred culture were higher than those in static culture, (p < 0.05). The primitive cells (CD34(+), CD34(+)/CD38(-) or CD133(+)) of the expanded cells in magnet stirred culture were less than those in static culture (p < 0.05). However, the CD184(+) or CD62L(+) expanded cells were more than that in static culture (p < 0.05). It is concluded that magnet stirred culture favors the expansion of hematopoietic progenitor cells. The results will be finally confirmed in further in vivo experiments and clinical applications.

The aim of this study was to investigate the effect of human bone marrow mesenchymal stem cells on human T-cell proliferation resulted from stimulation with PHA and possible immunomodulating mechanism. T cells were positively selected by CD3(+) magnetic beads, and were then co-cultured with irradiated MSCs overnight before the addition of PHA. T-cell proliferation was measured by BrdU assay and the degree of apoptosis was assessed by flow cytometry with Annexin V/PI. T cells co-cultured with or without MSCs were treated with PHA for 72 hours, then harvested. They were labeled with anti-CD4, anti-CD8, anti-CD25 antibodies and analyzed by flow cytometry. The results showed that MSCs inhibited T-cell proliferation, but did not induce T cell apoptosis. There were no significant changes in the ratio of CD4(+) and CD8(+) T cells of MSC-treated group, as compared with the control group. After stimulation with PHA, there was an increase in CD4(+) T cells and decrease of CD4(+)CD25(+) cells in MSC co-cultured group. It is concluded that the MSCs inhibit T-cell proliferation after stimulation with PHA, and show more inhibitive effects on CD8(+) and CD4(+) T cells, but CD25(+) regulatory T cells may not be involved in this process.