OBJECTIVES: The core pathology of osteoporosis lies in bone resorption exceeding bone formation; thus, promoting osteogenesis is a key therapeutic strategy. The osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) forms the biological basis of bone formation. Astragaloside IV (A-IV), a major active component of Astragalus membranaceus, is known to enhance osteogenesis, but its precise molecular mechanisms remain unclear. This study aims to investigate the effects of A-IV on the proliferation and osteogenic differentiation of BMSCs from osteoporotic rats and to elucidate its molecular mechanism through the regulation of microRNA-21 (miR-21) and Notch2 expression. METHODS: After 1 week of adaptive feeding, mature female SD rats were randomly divided into a sham-operated (Sham) group (n=4) and an ovariectomized (OVX) group (n=8) to establish an osteoporosis model. Twelve weeks after surgery, BMSCs were isolated from femoral bone marrow and cultured. Cells were divided into a S-BMSCs group (from Sham), an O-BMSCs group (from OVX), and an A-BMSCs group (from OVX-derived BMSCs treated with A-IV). S-BMSCs and O-BMSCs were induced for osteogenic differentiation using osteogenic induction medium, whereas A-BMSCs were treated with A-IV before induction. Flow cytometry was used to identify mesenchymal stem cell surface markers (CD29) and hematopoietic stem cell marker (CD34) to confirm BMSC characteristics. Cell proliferation was assessed using the methyl thiazolyl tetrazolium (MTT) assay. Alizarin red staining was performed to quantify calcium nodule formation, and alkaline phosphatase (ALP) activity assays were used to evaluate osteogenic differentiation. Real-time reverse transcription PCR (real-time RT-PCR) was used to detect changes in osteogenic-related genes, runt-related transcription factor 2 (Runx2) and osteopontin (OPN), as well as miR-21 expression. Western blotting was performed to assess Runx2, OPN, and Notch2 protein expression. RESULTS: Flow cytometry confirmed that O-BMSCs retained the phenotypic characteristics of mesenchymal stem cells. A-IV significantly enhanced the proliferation of BMSCs from osteoporotic rats (P<0.05), increased ALP activity, and upregulated the mRNA and protein expression of Runx2 and OPN (P<0.05). Bioinformatic and experimental analyses demonstrated that miR-21 directly targeted Notch2. A-IV treatment increased miR-21 expression while suppressing Notch2 protein expression and inhibiting activation of the Notch signaling pathway (P<0.05). CONCLUSIONS Astragaloside IV promotes the osteogenic differentiation of BMSCs derived from osteoporotic rats by upregulating miR-21 expression and inhibiting the key Notch signaling protein Notch2, thereby relieving the Notch2-mediated suppression of osteogenesis.
Animals ; Triterpenes/pharmacology* ; Saponins/pharmacology* ; Osteogenesis/drug effects* ; MicroRNAs/metabolism* ; Rats, Sprague-Dawley ; Female ; Cell Differentiation/drug effects* ; Mesenchymal Stem Cells/drug effects* ; Signal Transduction/drug effects* ; Osteoporosis/pathology* ; Rats ; Cells, Cultured ; Receptor, Notch2/metabolism* ; Receptors, Notch/metabolism* ; Ovariectomy ; Cell Proliferation/drug effects*

Objective To observe the reception of using pig bone marrow stromal cells (BMSCs) that were transfected with human tissue factor pathway inhibitor (TFPI) to resolve the dysregulation of coagulation after liver xenotransplantation.Methods Pig BMSCs were immortalized by transfection with lentivirus containing SV40T and then transfection with human TFPI.At last the cells were tested for their ability to inhibit clotting in a model by co-incubation of human plasma,human monocytes and pig aortic endothelial cells.Results After transfection with SV40T,pig BMSCs were immortalized and similar to primary cells.The immortalized pig BMSCs showed a stable TFPI expression after transfection with human TFPI by lentivirus.Moreover,they showed the potential of regulating coagulation dysregulation in vitro.Conclusion Pig BMSCs transfected with human TFPI could solve the regulation dysregulation caused by TF activation effectively,and have the potential of resolving coagulation dysregulation in liver xenotransplantation.

Induction of immune tolerance is characterized by high efficiency,low toxicity and economic advantages,and immune tolerance has very important significance in organ transplantation.In the establishment of immune tolerance,dendritic cells (DCs) play the key roles such as clonal deletion or clonal anerge,expressing t-cell inhibiting factors,activating helper T cells electively and inducing regulatory T cells,especially CD4 + CD25 + regulatory T cells,etc.Similarly,CD4 + CD25 + regulatory T cells whose primary target point is dendritic cell also have various functions such as affecting dendritic cells'maturing,restraining dendritic cells'antigen presentation to T cells,inhibiting the activation of T cells in non-specific ways and inducing immune tolerance.Current research shows,dendritic cells and CD4 + CD25 + regulatory T cells regard each other as target point and work synergetically.Here we mainly discuss the function and interaction of dendritic cells and CD4 + CD25 + regulatory T cells in the immune tolerance.

[Objective]To investigate the metaphase result of extensive fusion in treatment of spondylolisthesis combined with intervertebral disc degeneration of adjacent joints.[Methods]Seventy-nine cases of lumbar spondylolisthesis combined with intervertebral disc degeneration of adjacent joints were treated with extensive fusion and internal fixation from September 2002 to September 2007.JOA and VAS scores were assessed and compared pre-and post-operatively.X-ray and CT were performed to observe the stabilization of fusion joints,and satisfaction rate from patients was investigated.[Results]All patients were followed up for 2 to 6.5 years(mean,4.6 years).Post-operative JOA scores increased(P

OBJECTIVETo study the optimum acupuncture treatment program and the mechanism for treatment of cerebral infarction.

METHODSSixty-three cases were randomly divided into 9 groups. The four factors, times of manipulation, the retaining time of the needle, acupuncture instrument and acupoints,and their corresponding three levels were adopted respectively in treatment of each group. Then nerve function defect score and insulin resistance were observed before and after treatment.

RESULTSThe acupoints, the times of manipulation and the retaining time of the needle have significant effects on nerve function defect and insulin resistance (P < 0.01 or P < 0.05), and the acupuncture instrument has a significant effect on insulin resistance (P < 0.01). The choice of acupoints was the most important factor for acupuncture treatment of cerebral infarction.

CONCLUSIONAcupuncture for regulating The Governor Vessel with twice manipulations and retaining the needle for 60 min is optimum treatment program for cerebral infarction. The good regulating effect of acupuncture on insulin resistance is one of the mechanisms of achieving the therapeutic results.

Acupuncture Points ; Acupuncture Therapy ; Cerebral Infarction ; Humans ; Insulin Resistance ; Needles