Screening and cloning of multi-drug resistant genes in HL-60/MDR cells.

Objective: To screen and clone multi-drug resistance (MDR) related genes in MDR acute myeloid leukemia cells (HL-60/MDR).

Methods: HL-60/MDR was established using All-Trans Retinoic Acid. With the HL-60 cells as "tester" and HL60/MDR as "driver", the cDNA library of HL-60/MDR was established by suppression subtractive hybridization.

[Role of the sonic hedgehog pathway in regulation of the proliferation, migration and differentiation of hemangioblast derived from AGM].

Objective: To explore the role of sonic hedgehog (Shh) pathway in regulating the proliferation, migration and differentiation of hemangioblasts derived from aorta-gonad-mesonephros (AGM).

Methods: The hemangioblasts were isolated from AGM region of 11-day postcoitum (dpc) murine embryos by using the immuno-magnetic with CD34 and Flk1 monoclonal antibodies. The phenotypic analysis of hemangioblasts and AGM-derived stromal cells were detected by flow cytometry.

[Establishment of multivariate diagnosis and treatment system of modern gynecology of traditional Chinese medicine].

Gynecology of traditional Chinese medicine (TCM) is a clinical subject concentrating on women's specific diseases. With the advance of the time, clinical gynecology of TCM has to be developed from clinical practice and establish an open mode of multivariate diagnosis and treatment. The author of this article expatiates on the relationships between syndrome differentiation and disease identification, constitution differentiation and syndrome differentiation, stage differentiation and syndrome differentiation, time differentiation and syndrome differentiation, macroscopic differentiation of syndrome and microcosmic differentiation of syndrome, and integral syndrome differentiation and local syndrome differentiation, and then states that it is necessary for us to establish a multivariate diagnosis system to adapt to the complex clinical practice.

Screening of aplastic anaemia-related genes in bone marrow CD4+ T cells by suppressive subtractive hybridization.

Background: CD4(+) T cells play a crucial role in the pathogenesis of aplastic anaemia. However, the mechanisms of over-proliferation, activation, infiltration of bone marrow and damage to haematopoietic cells of CD4(+) T cells in aplastic anaemia are unclear. Therefore, we screened differentially expressed genes of bone marrow CD4(+) T cells of aplastic anaemia patients and normal donors by suppressive subtractive hybridization to investigate the pathogenesis of aplastic anaemia.

[Mechanism of radiation induced premature senescence of bone marrow stromal cells: experiment with murine bone marrow stromal cells].

Objective: To investigate the mechanism of radiation induced premature senescence of bone marrow stromal cells induced by gamma-radiation.

Methods: Bone marrow stromal cells were isolated from the bones of mice, cultured, and divided into 2 groups: control group and irradiation group. The cells of the irradiation group were exposed to 60Co gamma-rays of the dose of 8.

[Screening and cloning the genes differentially expressed in human embryonic AGM-derived stromal cells].

To screen and separate the genes differentially expressed in human embryonic aorta-gonad-mesonephros (AGM)-derived stromal cells, a subtracted library was generated through the suppression subtractive hybridization using the cDNA of human embryonic AGM-derived stromal cells as target and human fetal liver (FL)-derived stromal cells as drivers. Then a high though screening technique, gene chip, was used to screen the differentially expressed genes in the established subtractive library. Approximately 18 of the resulting subtracted cDNA clones were partially sequenced and analyzed by blastn in the GenBank database.

Sonic hedgehog protein promotes bone marrow-derived endothelial progenitor cell proliferation, migration and VEGF production via PI 3-kinase/Akt signaling pathways.

Aim: To investigate the effects of Sonic hedgehog (shh) protein on bone marrow- derived endothelial progenitor cells (BM-EPC) proliferation, migration and vascular endothelial growth factor (VEGF) production, and the potential signaling pathways involved in these effects.

Methods: Bone marrow-derived Flk-1(+) cells were enriched using the MACS system from adult Kunming mice and then BM-EPC was cultured in gelatin-coated culture dishes. The effects of shh N-terminal peptide on BM-EPC proliferation were evaluated using the MTT colorimetric assay.

An experimental study on astrocytes promoting production of neural stem cells derived from mouse embryonic stem cells.

Background: The production of neural stem cells (NSCs) derived from embryonic stem (ES) cells was usually very low according to previous studies, which was a major obstacle for meeting the needs of clinical application. This study aimed at investigating whether astrocytes could promote production of NSCs derived from ES cells in vitro.

Methods: Mouse ES cells line-D3 was used to differentiate into NSCs with astrocytes as inducing stromal cells by means of three-stage differentiation procedure.

Expansive effects of aorta-gonad-mesonephros-derived stromal cells on hematopoietic stem cells from embryonic stem cells.

Background: Hematopoietic stem cells (HSCs) give rise to all blood and immune cells and are used in clinical transplantation protocols to treat a wide variety of refractory diseases, but the amplification of HSCs has been difficult to achieve in vitro. In the present study, the expansive effects of aorta-gonad-mesonephros (AGM) region derived stromal cells on HSCs were explored, attempting to improve the efficiency of HSC transplantation in clinical practice.

Methods: The murine stromal cells were isolated from the AGM region of 12 days postcoitum (dpc) murine embryos and bone marrow (BM) of 6 weeks old mice, respectively.

[The effects of Mcl-1 gene on ATRA-resistant HL-60 cell].

Objective: To investigate the role of Mcl-1 gene in resistance of all-trans retinoic acid (ATRA) of leukemia cells.

Methods: Long-term, intermittent and repetitive exposure of HL-60 cells to ATRA was used to establish a multidrug-resistance cell line (HL-60/ATRA). HL-60/ATRA cells were transfected with Mcl-1 small interference RNA (siRNA) by Lipofectamine 2000.