To observe the effect of fuzheng yangying oral solution on bone marrow proliferation and serum interleukin 2 in immune-induced aplastic anemia mice.

To observe whether HSP70 could protect against H2O2-induced apoptosis in C2C12 myogenic cells by inhibiting Smac release from the mitochondria.

To perform the proteome analysis of conditioned medium prepared from mouse embryonic fibroblast feeder layers by 2-dimensional (2D) electrophoresis and mass spectrometry and to find out the possible differentiation-inhibitory factor in conditioned medium.

To explore the mechanism of differentiation of bone marrow stromal cells (BMSCs) into neurons in different micro-environments in vitro.

In order to explore the transfection and expression of hRI gene on human umbilical blood stem cells, and observe it's effect on the tumor growth. After enriching human umbilical cord blood CD34+ cells with a high-gradient magnetic cell sorting system (MACS), transfected them with supernatant of retrovirus containing human Ribonuclease inhibitor (hRI) cDNA. Hematopoietic progenitor clonogenic assay and PCR were used to evaluate transfection efficiency, and Western-blot and immune fluorescence were used to evaluate the expression quantity of hRI gene after transfection. Observe the effect of RI on the growth of melonoma in B16C57BL mice. The results showed that human umbilical blood CD34+ cells were highly purified by MACS, which made the purity of human umbilical blood CD34+ cells average 96.15%. hRI can be transfected on umbilical blood CD34+ cells, and the transfection efficiency was 35%. The positive expression of hRI gene on transfected CD34+ cells is identified by Western-blot and immune fluorescence assay. Mice injected with transfected CD34+ cells show a significant restraint of the tumor growth, a lower efficiency of tumor formation, a lower weight of the tumor and a longer incubation period of tumor formation with respect to the control groups. The results demonstrated the capacity of RI to inhibited the tumor growth by blocking the vasculature in tumor.

To investigate the feasibility of transplanted endothelial progenitor cells transfected with VEGF165 gene to ischemic flap with increased neovascularization and augmented the survival areas.

To investigate the effect of real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) approach in chronic myeloid leukemia (CML) for detecting the minimal residual disease (MRD) or monitoring the treatment response and predicting the prognosis.

Recently, many researches indicated the important role played by homeobox (HOX) gene family in normal hematopoiesis. As a kind of transcription factors, HOX gene products regulate and control the expression of target genes by binding to special DNA sequences. HOXB, a member of HOX gene family, especially HOXB(4), interests people greatly. It has been found that its expression relates closely to the self-renewal of hematopoietic stem cells and effective proliferation of hematopoietic progenitor cells. This review presents some new research progress in this area.

Recombinant human granulocyte-colony-stimulating factor (rhG-CSF) can widely regulate human immunologic response. In the protocol of peripheral blood stem/progenitor cell mobilization, rhG-CSF can change the numbers and functions of T cells. Then the results can impact the incidence of graft-versus-host disease after allogeneic peripheral blood stem/progenitor cell transplantation. The regulation of rhG-CSF on T cell is an indirect action which is based on the direct action to monocytes and dendritic cells. The numerous IL-10 secreted by monocytes plays a key role in cytokines production, proliferative response and cytotoxicity of T cells. Endogenous IL-10 can induce high expression of SOCS3 and the SOCS3 is very important for regulating the signal transduction of the activities of T cells. In this review influences of rhG-CSF on T-cells in mobilization process and related mechanisms were elaborated with emphasis.

To study clinical outcome of G-CSF-mobilized allo-peripheral blood stem cell (PBSC) and allo-bone marrow (BM) transplantation for patients with leukemia, donors were injected G-CSF 8-10 microg/(kg.d) for 5 days, PBSC were collected on day 4-5 and G-CSF mobilized BM was extracted on day 7. Conditioning regimen consisting of fludarabine, busulfan and cyclophosphamide. The results showed that transplanted cells in all patients were engrafted, the median days of neutrophil exceeding 0.5 x 10(9)/L and platelet exceeding 20 x 10(9)/L were 10.2 days (range 9 - 12 days) and 12.5 days (range 12 - 14 days), respectively. Patients were monitored up to 100 days, 4 of 12 patients (33.3%) developed II aGVHD, 10 of 12 patients (83.3%) developed limited cGVHD. The median follow-up duration was 5 months. Two patients died, the others were alive in disease-free situation. In conclusion, allogeneic transplantation of G-CSF mobilized PBSC plus BM was safe and effective treatment for patients with leukemia, the therapy provides rapid and sustained engraftment without increase in incidence of aGVHt and cGVHD.

To evaluate the hematopoietic stem/progenitor cell apheresis effect of Cobe Spectra (Version 6.1) and Fenwal CS 3000 Plus cell separators, fourty-two procedures on twenty donors using Cobe Spectra cell separator and twenty-two procedures on sixteen donors using Fenwal CS 3000 Plus cell separator were retrospectively analyzed. The number of CD34(+) cells collected, the collection efficiency (CE) of CD34(+) cells and the contaminations of red blood cell and platelet in the stem/progenitor cell products of two devices were compared. The results showed that there were no significant differences in the total number of CD34(+) cells collected and the CD34(+) cell CE between the two devices. There were positive correlations between the count of peripheral blood cells including leukocyte, monocyte, hematopoietic progenitor cell and CD34(+) cell after mobilization and the total number of CD34(+) cells collected. The stepwise multiple variable analyses revealed the peripheral blood stem/progenitor cell count emerged as the only significant independent predictive factor for CE. A negative correlation was seen between the peripheral blood monocyte count and the CD34(+) cell CE for the Fenwal CS 3000 Plus. The Fenwal CS 3000 Plus product contained more red blood cells than that of the Cobe Spectra. The decrease in the peripheral platelet count after Fenwal CS 3000 Plus apheresis was also greater. It is concluded that collection efficacy of Cobe Spectra (Version 6.1) and Fenwal CS 3000 Plus was similar. Cobe Spectra shall be used preferably to assure higher CD34(+) cell CE at a high peripheral blood monocyte count. The Cobe Spectra cell separator is better for the donors with mismatched blood type and the donors with thrombocytopenia.

In order to research the related factors of umbilical cord blood transplantation, 54 cases of unrelated umbilical cord blood transplantation were analyzed retrospectively, which were performed from June 1998 to July 2003. All cord blood units were obtained from full term normal vaginal deliveries in Guangzhou Maternal-Neonatal Hospital. The fractionation, cryopreservation and thawing of cord blood have been done according to the regulation of New York umbilical cord blood bank and pertinent literature. The selection of cord blood is based on HLA typing and the number of nucleated cells. The results showed that from June 1998 to July 2003, 3 475 units of cord blood were collected in Guangzhou Umbilical Cord Blood Bank and 99 units were provided for therapy of 85 patients in 21 transplantation centers, including 11 sibling and 74 unrelated cord blood transplantations. 54 cases of unrelated cord blood transplantation were reported, including 43 malignant diseases and 11 non-malignant diseases. The median age of recipients was 9.5 (range 1.2 - 33) years, the median weight was 27 (range 10 - 60) kg, the median number of TNC was 6.82 x 10(7)/kg, 43 cord blood were implanted (ANC > 500/microl) at day 60 after transplantation (79.6%, median 17). The time of nuclear cell reconstitution after cord blood transplantation was statistically related with nucleated cells and the type of disease, not related with HLA matching. Acute GVHD was present in 8 patients (21.6%) and chronic GVHD occurred in 2 patients (5.4%), 6 patients suffered from graft failure (11.1%). The total survival rate was 42.6%. It is suggested that unrelated umbilical cord blood transplantation seems to be a good substitute for bone marrow transplantation and has good prospects especially in children.

The aim was to study the roles that the bone marrow mesenchymal stem cells (MSC) and cytokines play in cord blood CD34(+) cell expansion ex vivo and the influence of culture ex vivo on expression of the adhesive molecule of CD44. CD34(+) cells sorted from cord blood cells had been cultured in each well of 24 well culture plates containing culture medium supplemented with mesenchymal stem cells layer or/and cytokines for a week, and then all kinds of indexes of different groups were compared. The results showed that as for cord blood cell expansion, there was no significant difference between the groups with cytokines SDF-1alpha + SCF + TPO + FL and SCF + TPO + FL no matter if MSC layer existed or not. The groups with MSC layer and cytokines were superior to the corresponding groups without MSC layer. In addition, the expression of the adhesion molecule CD44 had no distinct change after culture. It is concluded that SDF-1alpha has no distinct influence on the effect of cytokines SCF + TPO + FL on cord blood cell expansion ex vivo. MSC enhance the effect of cytokines on cord blood cell expansion ex vivo. Such expansion ex vivo may not influence the expression of the adhesive molecule CD44 on cord blood cells.

To cultivate CD34(+)CD38(-) cells isolated from umbilical cord blood of healthy puerperal women over a longer-period of time for observation of cell division, proliferation, apoptosis, and effects of stem cell factor on the growth of CD34(+)CD38(-) cells, with flow cytometry, CD34(+)CD38(-) cells were isolated from umbilical cord blood of 10 healthy puerperal women and cultivated in stem cell media with supplement of IL-3, IL-6, GM-CSF, EPO, IGF-1 and SCF 6 kinds cell growth stimulating factors for six months. The cell growth curves were established. The effects of stem cell factor on the growth of CD34(+)CD38(-) cells and cell apoptosis were investigated with the single cell gel electrophoresis technique and flow cytometry method, respectively. The results showed that CD34(+)CD38(-) cells isolated from umbilical cord blood were capable of proliferating after being cultivated in vitro over a longer-period of time with no evidence of the presence of excessive apoptosis. In conclusion, under appropriate culture conditions, CD34(+)CD38(-) hematopoietic early progenitor cells from umbilical cord blood can serve as a resource providing a large amount of primitive cells for transplantation therapy after a longer period of cultivation and proliferation in vitro.

Embryonic stem cells are pluripotent and their differentiation in vitro can serve as an experimental model to explore the molecular mechanisms of early embryonic development. To investigate the effect of stromal cell conditioned medium combined with cytokines (sccm + cys) on the differentiation from human embryonic stem cells to hematopoietic cells and endothelial cells, the mouse fibroblast feeder cells to make human embryonic stem cells grown into embryonic bodies (EBs) were initially deleted. After culture for 3 days, EB cells were trypsinized into single cells and induced for 8 days by sccm + cys. Then, the differentiated cells were cultured in the semisolid medium containing 0.9% methylcellulose and cytokines to study the colony forming and self-renewal ability of cells. Immunocytochemical staining was used to check the surface markers of the colony cells. During the induction, mRNA expression of flk-1, BMI-1, scl, and Zeta-globin genes was tested by RT-PCR. Surface markers, such as flk-1, CD34 were tested by the flow cytometry. The results demonstrated that: (1) cell clusters containing 20-30 cells were formed after culture for 8 - 14 days in the semisolid medium, replanting these cells resulted in similar cell cluster forming. In addition, CD45 positive in big cell colonies were also found in the semisolid medium; (2) attached cell colonies appeared after culture for 8 days in the semisolid medium and VIII factor, UEA and KDR could be detected as negative by immunocytochemical staining; (3) on the 4(th) day of induction, mRNAs of flk-1, BMI-1, scl and Zeta-globin were all expressed. On the 8(th) day of induction, all of the above genes except Zeta-globin were expressed, while ES cell and EB cells which served as controls did not express scl and Zeta-globin genes; (4) on the 8(th) day of induction, the proportions of flk-1(+) cells and CD34(+) cells among all the inducing population were 9.8% and 16.8%, respectively, while the corresponding positive populations were 0.36% and 1.16% in spontaneously differentiated 11(th) day's EB, and 0.04% and 0.16%, respectively, in ES cells. If is concluded that embryonic stem cells can differentiate into hematopoietic cells and endothelial cells in combinant culture system of this study.

To investigate the differentiation of bone marrow derived Thy-1+ beta2M- cells (BDTCs) into liver cells in allyl alcohol (AA) induced liver injury micro-environment.

To investigate and compare the therapeutic effect of umbilical cord blood stem cell transplantation (UCBSCT) or adult fresh plasma in severe viral hepatitis liver failure with/without heart damage, and to study the effect of UCBSCT on liver lesions in rats.

Nuclear transfer and therapeutic cloning have widespread and attractive prospects in animal agriculture and biomedical applications. We reviewed that the quality of oocytes and nuclear reprogramming of somatic donor cells were the main reasons of the common abnormalities in cloned animals and the low efficiency of cloning and showed the problems and outlets in therapeutic cloning, such as some basic problems in nuclear transfer affected clinical applications of therapeutic cloning. Study on isolation and culture of nuclear transfer embryonic stem (ntES) cells and specific differentiation of ntES cells into important functional cells should be emphasized and could enhance the efficiency. Adult stem cells could help to cure some great diseases, but could not replace therapeutic cloning. Ethics also impeded the development of therapeutic cloning. It is necessary to improve many techniques and reinforce the research of some basic theories, then somatic nuclear transfer and therapeutic cloning may apply to agriculture reproduction and benefit to human life better.

To investigate the effectiveness of human insulin-like growth factor I (hIGF-I) gene transferred into the cultured goat bone marrow mesenchymal stem cells with liposome, and find a new method of cell-mediated gene therapy.

To study an efficient method to transfect green fluorescent protein gene (GFP) to rat bone marrow mesenchymal stem cells (MSCs) and to determine the biological properties and differentiation potency of transfected MSCs.