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Objectives:This study aims to analyze the current status of the collaboration of medical staff among county,township,and village with comprehensive prevention and control of chronic diseases based on nine dimen-sions of DMIC integrated medical service development model,with a view to providing medical services collaboration on chronic diseases across the organization to provide direction for improvement. Methods:278 health workers sam-pled by the combination of typical sampling and multi-stage random sampling method received a questionnaire and the results were analyzed statistically. Results:According to the findings of this study,village doctors show higher partic-ipation in chronic care collaboration,but they are faced with the aging population and most of them are poorly educat-ed;the multi-institutional collaboration is of low efficiency in practice although it is well accepted by all health work-ers;there is no existing shared health information system among county,township and village. Conclusion:These re-sults suggest that the government should strengthen its leadership and promote primary health care management through medical staff collaboration;initiatives of health workers should be stimulated to promote the continuity of medical serv-ices;the shared health information system should be set up to facilitate the health workers' collaboration.
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<p><b>OBJECTIVE</b>To investigate the effect of myeloma-derived exosomes on surface activating receptors of NK cells, and to explore the mechanism of the function defect of NK cells.</p><p><b>METHODS</b>The exosomes from the supernatant of multiple myeloma cell lines RPMI8226 and U266 were extracted by ultracentrifugation, and the size of them was identified under electron microscope; the human primary NK cells were extracted, and were co-cultured with the myeloma-derived exosomes (40 µg/ml), then the expression levels of surface activating receptors NKp46, NKp30 and NKG2D of NK cells at 0,1,4 and 24 hours were detected by flow cytometry.</p><p><b>RESULTS</b>The exosomes showed small vesicular, sized 30-100 nm under electron microscope. The expression of surface activating receptors of NK cells declined at different degree after co-cultured with myeloma-derived exosomes.</p><p><b>CONCLUSION</b>Myeloma-derived exosomes can inhibit the expression of surface activating receptors of NK cells.</p>
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ObjectiveTo investigate the gene expressions of TAZ and Wnt/β-catenin on the postosteogenic cells of mesenchymal stem cells(MSC)in multiple myeloma(MM)patients and to explore the potential therapeutic target of multiple myeloma bone disease (MBD).MethodsBone marrow mononuclear cells MNC from MM and controls were isolated,cultured,expanded and then induced to osteogenic differentiation.Realtime quantitative RT-PCR was employed to detect the osteogenic markers (TAZ,Wnt/β-catenin,OPN,OC,ALP and Cbf α1); and alizarin red staining for mineral deposition.The mRNA expressions of TAZ and Wnt/β-catenin in the two groups were analysed.ResultsAlizarin red staining was positive and the red calcium nodules were appeared on the post-osteogenic cells of MSC.The mRNA expressions of OC,ALP and Cbf α1 were 2.0958±0.5665,2.6670±0.3847,0.8463±0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly higher than those of pre-osteogenic cells(1.3487±0.9291,1.1452±0.6054,0.4439±0.2945) (t =2.171,6.709,2.795; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP and Cbf α1 were 2.1096±0.8267,2.8991±0.3531,4.3045±0.2844,1.3273±0.4075,respectively,on the post-osteogenic cells of MSC in the controls,which were significantly higher than those of pre-osteogenic cells (1.2200±0.9091,0.8780±0.3927,1.9161±0.2684,0.6736±0.2513) (t =2.289,12.103,25.134,4.411; all P ＜ 0.05).The mRNA expressions of OPN,OC,ALP,Cbf α1 were 1.2710±0.5636,2.0958±0.5665,2.6670± 0.3847,0.8463+0.3473,respectively,on the post-osteogenic cells of MSC in the experimental groups,which were significantly lower than those of control groups(2.1096 ±0.8267,2.8991 ±0.3531,4.3045±0.2844,1.3273±0.4075) (t =-2.650,-3.805,-10.822,-2.841; all P ＜ 0.05).The mRNA expression of TAZ and β-catenin were 2.2315±1.0723 and 0.5801±0.2159 on the post-osteogenic cells of MSC in MM patients,which were significantly lower than those of control groups (4.4140±0.8325,0.9516±0.2920) (t =±5.085,-3.235;both P ＜ 0.05).ConclusionThe gene expressions of OPN,OC,ALP and Cbf α1,the osteogenesis related genes,are increased in post-osteogenic cells of MSC,which showed the MSC have been successfully induced to osteoblasts.Comparing with control groups,the osteogenic potential of MSC in MM patients is lower.Based on the above research,TAZ and Wnt/β-catenin may present a novel target for the future therapy of MBD.
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Objective To investigate the reversal effect of Topo Ⅱα-shRNA and Topo Ⅱβ-shRNA on Topo Ⅱ gene in K562/AS2 cells. Methods Three pieces of Topo Ⅱα-shRNA and three pieces of Topo Ⅱβ-shRNA were designed,synthesized and transfected into K562/AS2 cells by liposome. Expression level of Topo Ⅱα and Topo Ⅱβ mRNA were determined by real time fluorescence quantitative PCR. The expressions of Topo Ⅱα and Topo Ⅱβ protein were assayed with flow cytometer. Results After treated with Topo Ⅱα-shRNA or Topo Ⅱβ-shRN A for 24 hours,the expression level of Topo Ⅱα mRNA and Topo Ⅱβ mRNA protein in K562/AS2 cells decreased at most (78.22±0.01) %,(31.17±1.27) % (P <0.05),and (57.36±0.01)%,(23.98±1.22) % (P <0.05) respectively. Conclusion The expression of Topo Ⅱ gene can be down-regulated after infected with Topo Ⅱ-shRNA in K562/ AS2 cell line.
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Objective To investigate sequence variations of 12 miRNA genes in multiple myeloma(MM) in order to find whether sequence variations in miRNA genes are associated with tumorigenesis and discuss the clinical significance of MM associated with miRNA genes mutations. Methods The miRNA gene mutations in 20 cases of MM, 4 MM-derived cell lines and 20 controls were detected by the methods of polymerase chain reaction single stranded conformation polymorphism (PCR-SSCP) and silver staining technique. Both clinical features and laboratory results were analyzed simultaneously. Results The electrophoretic patterns showed a total of three variations in miR-19a, miR-19b and miRNA-335,which were observed in 3 MM cells (15 %, 3/20). We also found variations of miRNA-335 in MM-derived cell lines KM-3and RPMI8226. However, no sequence alteration in the miRNA genes was observed in our set of controls. One of the three MM patients died, and two of them were detected mutations at the terminal stage of the disease.Conclusion A relative high frequency of miRNA gene mutation was found in MM and MM derived cell lines, which suggests possibility of a main mechanism underlying tumorigenesis. And, detecting miRNA gene mutations in MM might be benefit to evaluate the progression and prognosis of disease.
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Objective To construct a recombinant adenovirus vector of Hairpin RNA specific for MRP1 gene and study its inhibition of MRP1 gene expression in K562/AS2 cell line resistant to AS_2O_3 (ATO). Methods A MRP1-specific hairpin RNA recombinant adenovirus vector was constructed and used to infected K562/AS2 cells. Expression level of MRP1 mRNA detected by real-time fluorescent quantitative PCR. MRP1 protein detected by flow cytometry. MTT method was used to detected the cytotoxicity of ATO and etoposide. Results MRP1 mRNA and protein expression level in K562/AS2 cells before and after the pAd-MRPl-shRNA adenovirus infection was (34.70±0.28 vs 4.19±0.03, P <0.05) and (26.40±0.16 vs 10.85±0.37, P<0.05), respectively. RR of K562/AS2 to arsenic trioxide and etoposide was (11.4078±0.3183 fold vs 1.6126±0.3015 fold, P<0.05) and (5.9141 ±0.0149 fold vs 1.7664±0.1038 fold, P <0.05), respectively. The reversal fold of ATO and etoposide was (7.2409±1.3668) and (3.3555±0.1886), respectively. Conclusion Successfully constructed pAd-EGFP-U6-shRNA-MRPl adenovirus vector, the vector of infection K562/SA2 cells can inhibit MRP1 gene expression and reverse the resistance of the ATO and etoposide.
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Objective To evaluate the influence of tissue factor pathway inhibitor 2(TFPI-2)gene on the proliferation and invasion of K562.Methods The expression vector pcDNA3.0/TFPI-2 was transfected into human leukemia line K562 cells(K562-T)by using liposome,then the mRNA and protein TFPI-2 were detected by real-time RT-PCR and western blot separately.The growth curve and the colony-forming unit assay were used to measure the ability of cells growth and transwell chamber model was employed to test the ability of cell invasion in vitro.Results Expression of mRNA and protein of TFPI-2 was detected in transfected cells.The growth rate and self-replication ability of K562-T cells were lower than those of the two control groups obviously.The number of K562-T cells to traverse a Matrigel-coated membrane was dramatically decreased compared with that of non-expressing cells.Conclusion The gene of TFPI-2 can inhibit the growth,proliferation and invasion of the K562 cells.
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<p><b>OBJECTIVE</b>The radial artery differs from internal mammary artery in its vascular biology and long-term patency after coronary artery bypass grafting (CABG). This study was designed to investigate their ultrastructural differences that may have implications in arterial remodeling and graft failure.</p><p><b>METHODS</b>Thirty-four radial artery and 11 internal mammary artery samples were obtained from patients underwent CABG, and subjected to routine electron microscopic examination. A semi-quantitative method was used to evaluate secretary endothelial cells, endothelial denudation, synthetic smooth muscle cells (SMCs), matrix accumulation, lipid deposition and medial submicroscopic calcification.</p><p><b>RESULTS</b>Compared with internal mammary arteries, the radial arteries had more secretory endothelial cells (47.1%, 16/34 vs 27.2%, 3/ 11) and synthetic type SMCs in a background (14.4% vs 0.9%) that had more intimal lipid deposition and matrix accumulation (14.7%, 5/34 vs 9.1%, 1/11). Matrix vesicles and calcifications were frequently present in the media of both types of arteries. The calcifications, however, could not be visualized by routine histological stains, and therefore, named as submicroscopic calcification in this study. Fewer endothelial denudations were observed in the radial arteries, but no differences in medial lipid deposition and submicroscopic calcification were observed between these two types of arteries. The ultrastructural features and the arrangement of medial SMCs in radial arteries were similar to those of internal mammary arteries.</p><p><b>CONCLUSIONS</b>Radial arteries have a higher SMC proliferative potential and more actively secretory status of endothelial cells, which may enhance the remodeling process and correlate with a decreased long-term patency. Better preservation of endothelial cells in radial arteries could be attributed to the "no touch" technique utilized in surgical harvesting. The significance of submicroscopic medial calcification during graft remodeling requires further investigations.</p>