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ObjectiveTo explore the potential mechanism of Polygonati Rhizoma on the treatment of osteoporosis （OP） based on network pharmacology and molecular docking method and to verify the mechanism by experiments. MethodThe main active ingredients and corresponding targets of Polygonati Rhizoma were screened out from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） 2.3 by conditional searching. The treatment targets were obtained from the genes related to OP and DisGeNET 7.0. The potential target genes of Polygonati Rhizoma for treating OP were obtained by the crossing of the corresponding targets and the treatment targets. Cytoscape 3.7.1 was used to construct the “Polygonati Rhizoma-active ingredient-potential target” network. The protein-protein interaction （PPI） analysis was carried out by STRING 11.0， and the PPI network was constructed. Metascape 3.5 was used to conduct Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses of the key targets. The core ingredients and key targets of Polygonati Rhizoma were selected for molecular docking by AutoDock Vina 1.1.2. Finally， the effect of β-sitosterol on osteogenic differentiation of MC3T3-E1 cells in rats was observed. ResultTwelve active ingredients and 32 potential targets of Polygonati Rhizoma for OP treatment were screened out. Six active ingredients including baicalein and β-sitosterol and key targets including protein kinase 1 （Akt1）， tumor suppressor p53 （TP53）， vascular endothelial growth factorA （VEGFA）， proto-oncogene Jun （JUN）， matrix metalloproteinase-9 （MMP-9）， and proto-oncogene c-Fos （FOS） were obtained by Cytoscape 3.7.1 topological analysis. A total of 995 GO entries and 181 signaling pathways involving the response to reactive oxygen species and regulations of growth were obtained from GO and KEGG enrichment analyses. The results of molecular docking showed that the core active ingredients possessed good binding activities with the respective key targets. The results of cell experiments showed that β-sitosterol promoted the osteogenic differentiation at the concentration of 2.5 μmol·L-1 and 5 μmol·L-1. ConclusionPolygonati Rhizoma had the therapeutic effect on treating OP by regulating inflammation， oxidative stress， apoptosis， and metabolism. The β-sitosterol significantly promoted the osteogenic differentiation of MC3T3-E1 cells.
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<p><b>OBJECTIVE</b>To explore the treatment skill and clinical effect of bone setting manipulation in treating proximal humeral fracture with shoulder dislocation.</p><p><b>METHODS</b>From January 2015 to December 2015, 118 cases of proximal humeral fractures with shoulder joint dislocation were treated by bone setting reduction and fixation with adhesive tape or Kirschner wire after reduction, including 56 males and 62 females with an average age of 61 years old ranging from 48 to 88 years old. The time from injury to treatment was 1 to 31 days with an average of 3 days. All fractures were closed, according to Neer standard classification, there were 27 cases of type II, 55 cases of type III and 36 cases of type IV. There were 85 cases of shoulder subglenoid dislocation, 33 cases of subcoracoid dislocation. All the patients were accompanied by obvious shoulder joint deformity, dysfunction and other symptoms. All cases were diagnosed by X-ray examination, and the bone setting manipulation was performed under the brachial plexus anesthesia. According to the type of fracture and fixation method, the patients were encouraged to exercise the limb rehabilitation, and the curative effect was determined by Constant-Murley shoulder score system.</p><p><b>RESULTS</b>All cases were followed up for 6 to 12 months with an average of 9 months. There were 1 case of humeral head necrosis, 2 cases of traumatic frozen shoulder. The score of shoulder joint function Constant-Murley was 84.5±4.5; among them, 78 cases were excellent, 28 cases were good, 9 cases were fair, 3 cases were poor, the excellent and the good rate was 89.8%. X-ray showed bony union of all cases.</p><p><b>CONCLUSIONS</b>The treatment of proximal humeral fracture with dislocation of shoulder joint by bone setting manipulation is a skillful method, with simple operation, small trauma and satisfactory curative effect.</p>
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Objective To investigate the effect of neuropeptide Y (NPY) on the osteoblastic differentiation of murine MC3T3-E1 cells and its mechanism related to the Wnt signaling pathway.Methods The murine MC3T3-E1 cells were divided into 4 groups according to the stimulators added:phosphate buffered saline (PBS) (control) and different concentrations of NPY (10-8 mol/L,10-10 mol/L and 10-12 mol/L).The cellular proliferation was detected with MTT assay after 1,3,5,7 and 9 days.The cells were identified with cell immunochemistry and Western Blot to find out the most effective concentration of NPY at different time points under osteoblastic condition.The cells were then divided into 4 groups:PBS,NPY,NPY + NPY receptor antagonist,and NPY + DKK1.Western blot was used to determine the expression of β3-catenin and p-GSK-3β in each group.Nuclear signaling activity of β3-catenin was observed using immunofluorescence staining.Results NPY significantly improved the proliferation of MC3T3-E1 cells at 7 and 9 days (P ＜0.05).NPY (10s mol/L and 10-10 mol/L) groups and NPY (10-10 mol/L and 10-12 mol/L) groups significantly improved the ALP activity at 4 and 14 days respectively (P ＜ 0.05).At 4 days,the expression of ALP protein was significantly decreased in the NPY + DKK1 group and the NPY + NPY receptor antagonist group compared with that in the NPY group (P ＜ 0.05).Although the expression levels of [β-catenin and p-GSK-3β protein were uninfluenced in either case,NPY significantly stimulated the nuclear signaling activity of β3-catenin.Conclusions NPY may significantly increase the expression of ALP protein in MC3T3-E1 ceils during osteoblastic differentiation.This effect might be mediated through the canonical Wnt signaling pathway.
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Objective To supply large amount of seed cells for the cartilaginous tissue engineering,adult adipose-derived stem cells(ADSCs) were differentiated directly into cartilaginous cells.Methods Adipose tissue donated voluntarily by the adult patients after liposuction was isolated and subcultured.The cell activity was detected by methyl thiazolyl tetrazolium(MTT) assay and the proliferation curve line was drawn.The cultured stem cells were identified with flow cytometry.After the subculturing,the fourth generation of ADSCs were differentiated directly into cartilaginous cells,and then were marked by Alcian blue.Type II collagen expression in the ADSCs after cartilaginous differentiation was examined by immunohistochemical assay.Results ADSCs proliferated in vitro fast,and kept a stable multiplication till the 13th~15th generation.The results of flow cytometry showed that the cultured ADSCs had the specific features of the stem cells,and then were differentiated into cartilaginous cells.Positive Alcian blue staining and type II collagen expression indicated that the obtained cells had the function of normal cartilaginous cells.Conclusion ADSCs are differentiated directly into cartilaginous cells successfully,which will supply seed cells for the cartilaginous tissue engineering.