To detect the quantity, proportion and function of producing cytokines of Th1 and Th2 cells in aplastic anemia (AA) patients and their contribution to the hematopoietic failure.

To construct the retroviral expression vector of BALB/c mouse H-2Dd gene and study its expression in C57BL/6 mouse hematopoietic cells (MHC).

This study was designed to evaluate the effectiveness of high-dose chemotherapy plus single daily dose of 250 micrograms/d rhG-CSF(lenograstim) in autologous peripheral blood stem cells mobilization and hematopoietic reconstitution after transplantation.

To evaluate the effect of calcification of autogenous bone marrow stem cell transplantation in periodontal tissue regeneration.

Since the diagnostic criterion of acute biphenotypic leukemia(BAL) was recommended by European Group for the Immunological Characteristics of Leukemia (EGIL) in 1995, the reports on cytogenetic feature of BAL can be seen in homeland and outside, but the case numbers in the reports were low. This study was designed to explore the cytogenetic feature of BAL.

Objective. To examine the effect of electromagnetic fields (EMF) (20 Hz, 8 mT; 5 Hz, 8 mT) on the neuron-orientated differentiation of neural stem cells (NSCs) from midbrains of new-bom rats. Method. Differentiated NSCs were exposed to EMF for 2 x 15 min per day lasting for I d, 5 d, or 10 d. The sham-exposure controls were correspondingly established. Cells were fixed and processed for immunofluorescent staining using the antibody against neuron-specific marker MAP2, then the percentage of MAP2+ cells was calculated. Result. The two EMFs promoted the differentiation of a neuronal fate in different ways. Both of them came into effect even after 1 d exposure. When cells exposed to the 20 Hz EMF, the percentage of neuron-orientated cells gradually increased with longer-term exposure and the most significant effect appeared in 10 d group while that happened in 5 d group under the condition of 5 Hz EMF. The effect contrasted horizontally, significant differences between the two EMFs were observed only at 10 d groups, 20 Hz EMF having more favorable effect than 5 Hz EMF. Conclusion. 20 Hz and 5 Hz EMF could promote the differentiation of midbrain NSCs to a neuronal phenotype in different ways, suggesting that the physical induction might be another strategy to manipulate the differentiation of NSCs.

We used the transplantation of purified autologous peripheral blood CD34+ stem cells to treat a 16-year-old female patient with systemic lupus erythematosus (SLE), who had received unsuccessful treatment with steroids and immunosuppressants, and has achieved satisfactory therapeutic effect. The diagnosis of SLE was established one year ago, and the patient had SLE Disease Activity Index (SLEDAI) of 21 on admission. After ineffective treatment with dexamethasone and cyclophosphamid (CTX) for 3 months, purified autologous peripheral blood CD34+ stem cell transplantation was adopted. Autologous peripheral hematopoietic stem cells were mobilized by intravenous injection of 2.0 g/d cyclophosphamid (CTX) for 3 d and subcutaneous injection of granulocyte colony-stimulating factor (G-CSF, 300 microgram/d). A CS-3000 plus blood cell separator was used to collect peripheral blood stem cells, and cell count of mononuclear cells and CD34+ stem cells and epitope analysis of T and B lymphocytes were performed by FACscan flow cytometry. After purification with CliniMACS, the number of CD34+ stem cells reached 15.13x106/kg, while that of CD3+ cells were only 1.35x105/kg. Pretreatment of the patient consisted of intravenous injection of (50 mg/kg each day)for 4 consecutive days and antithymocyte globulin (ATG, 2.5 mg/kg each day) for 3 consecutive days with methylprednisolone (MP) at the dose of 1.0 g on the first day and 0.5 g on the following 2 days. The granulocytes were recovered by G-CSF stimulation. The purified CD34+ stem cells (60 ml) were reinfused within 24 h after pretreatment, following which changes in clinical manifestations and immunologic markers were compared with those before the transplantation. Clinical and immunologic remissions were achieved after transplantation, with all the autoantibodies reversed to the negative, suggesting the short-term effectiveness of this therapy. Based on this observation, we conclude that this therapy is possible to effect an eventual cure of SLE in this case, but the long-term effect needs to be further observed in the follow-up study.

The iron response element (IRE) is a highly conserved RNA stem loop structure. It is the binding site of iron regulatory protein (IRP). IRP binding to IRE is regulated by cellular iron. When cells are derived of iron, IRP binds IRE. If IRE is located at 5'UTR, IRP binding will inhibit translation initiation, else if IRE is at 3'UTR, IRP binding will stabilize mRNA and prevent it from degradation. So far all known IREs have C at the 1 position and G at the 5 position of the loop (C1G5 type). In vitro studies suggest that the U1A5 type IRE, which has U and A at the 1 and 5 loop position respectively, binds well to IRP. However, U1A5 type's in vivo existence is still elusive. IRE-IRP binding is involved in the regulation of iron metabolism, oxidative stress and possibly aging. Here we use an improved computation method performing a comprehensive search of IRE in human and mouse genes. We try to catalog potential human and mouse IRE containing genes, at the same time identify potential U1A5 IREs.

To explore the isolation, purification and expansion of human umbilical cord blood mesenchymal stem cells (MSCs) into neuron-like cells in vitro.

To investigate the expression of mdr1 gene in hematopoietic cells of a murine bone marrow transplantation model and to explore the feasibility of transferring mdr1 gene into hematopoietic cells to pro-vent myelosuppression from chemotherapy.

In this article the author reviews the currently prevailing techniques in producing transgenic animals with recount of those employing microinjection, retrovirus infection, embryonic stem cells, sperm vectors and nuclei transfer of somatic cells. The prospective of these techniques are also dealt with in brief.

To detect the expression of human bone morphogenetic protein 7 (hBMP-7) mRNA in rabbit bone marrow-derived mesenchymal stem cells with hBMP-7 gene transfection mediated by retroviral vector.

To study the effects of alpha1,4Gal T antisense oligonucleotide mediated by lipofectin on human glioma cell line SWO-38.

The regulation of foreign gene expression in Insect-Baculovirus Expression System is very complex. In this report, the effect of 5'-UTR in the expression of hGH gene in cultured Sf9 cells was examined. A 18 bp length in the end of 5'-UTR of hGH (human Growth Hormone, hGH) cDNA including a stem-loop structure was deleted by PCR. The truncated hGH cDNA, delta 1hGH was cloned in pFastBac1, named pFast-Bac-delta 1hGH. After transforming into E. coli. DH10Bac, which have a shuttle vetor-Bacmid, the delta 1hGH was integrated into Bacmid by site-specific transposition, and an expression vector, rBacmid-delta 1hGH DNA was acquired. By transfecting the cultured Sf9 cells with the recombinant expression vector DNA, pure recombinant virus, rAcV-Bac-delta 1hGH was obtained, and hGH gene was expressed. Immuno-blot and Chemiluminescent assay revealed that the expressed hGH had normal immunological activity, the amount of hGH expression level in Sf9 cell supernatant infected with rAcV-Bac-delta 1hGH containing the truncated 5'UTR was four to five times higher than that infected with rAcV-Bac-hGH.

To understand the differential gene expression profiles between bone marrow cells and mobilized peripheral blood CD34(+) cells.

Embryonic stem cells are derived from inner cell mass of the preimplanted blastocyst or from primordial germ cells of the early embryos, with the capacity of unlimited growth and differentiation potential. Embryonic stem cells(ES cells) can differentiate into all kinds of cells and organs under proper condition. Due to this characteristics ES cells have the attractive prospect in basic research, transplantation and gene therapy.

This is a study on the cultivation condition in vitro and differentiation of neural stem cells from human embryonic brain in order to find a way to get purified multipotential neural stem cells. The single cells was derived from the three-month embryonic brain digested with trypsin, some cells was frozen, the other cells were expanded with EGF and bFGF, the single-cell-clone was obtained by the way of limited dilution, and the serum was used to induce the cells differentiation. The cells were detected with the method of immunohistochemistry. The results showed that a lot of neurospheres could be seen in the presence of mitogens (both EGF and bFGF) and serum could induce neural stem cells to differentiate into neurons, astrocytes, and oligodendrocytes. These indicate that the survival and proliferation of neural stem cells rely on the cooperation of EGF and bFGF. The neural stem cells can also be harvested from the frozen cells.

Serum-free murine bone marrow endothelial cell conditioned medium(mBMEC-cm) was collected. The mBMEC-cm was ultrafiltered in Centriprep. The concentrated retentive substance of mBMEC-cm (MW > 10,000) and filtrate of mBMEC-cm(MW < 10,000) were obtained. The effects of mBMEC-cm on the growth of CFU-E and BFU-E were investigated. The results showed: 1. The retentive substance of mBMEC-cm significantly enhanced the growth of CFU-E and BFU-E. The effects of mBMEC-cm(2%-6%, V/V) on the growth of CFU-E and BFU-E were dose-dependent. 2. The filtrate of mBMEC-cm markedly inhibited the growth of CFU-E and BFU-E. The effects of the filtrate of mBMEC-cm on the growth of CFU-E and BFU-E were also dose-dependent. The mechanism of regulation will be studied further.

By using the cultural method of CFU-F derived from bone marrow in the patient with acute leukemia in vitro, we dissected the effects of rhGM-CSF and rhG-CSF on growth of CFU-F derived from AL patient bone marrow. The results showed that rhGM-CSF and rhG-CSF enhanced the number of colonies of CFU-F and its proliferation activity. Both factors had a synergy on the above effects. The effects of rhGM-CSF on CFU-F were significantly greater than that of rhG-CSF. The data indicate that rhGM-CSF and rhG-CSF might regulate hemopoiesis more effectively by stimulating the growth of fibroblast and improving bone marrow microenvironment.

To evaluate the effect of curcumin on the proliferation of hematopoietic progenitor cells or leukemic stem cells.