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Objective: To explore the key genes and potential therapeutic drugs for osteoarthritis (OA) by bioinformatics.Method: The microarray data GSE55235 was downloaded from the data platform of gene expression omnibus (GEO) and the differentially expressed genes were screened by R language software (3.5.0).Then,the differentially expressed genes were subjected to gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway analysis with David online database.The protein-protein interaction was analyzed by String 10.5 online database and visual editing was analyzed by Cytoscape v3.6.1 software.Subnetwork module analysis was utilized by MCODE plugin to screen the core genes in the process of OA.Finally,small molecule drugs with potential treatment for OA were analyzed by connectivity map (CMap) database.Result: A total of 556 differentially expressed genes were screened,among which 252 were up-regulated and 304 were down-regulated.These genes were mainly involved in extracellular matrix (ECM) organization,inflammatory response,cell adhesion,immune response,collagen binding,etc.The analysis of KEGG pathway showed that differential genes were mainly involved in ECM-receptor interaction,phosphatidylinositol 3 kinase-protein kinase B (PI3K/Akt) signaling pathway and osteoclast differentiation.Some genes,such as interleukin-6(IL-6),JUN,vascular endothelial growth factor α(VEGFA),FOS,MYC and early growth response gene-1(EGR-1),activating transcription factor-3(ATF-3),playing critical role in the process of OA were identified by protein-protein interaction.Some potential small molecular drugs for the treatment of OA have also been screened,such as lycorine and anisomycin.Conclusion: The selected key genes may be targets for the diagnosis of OA or potential targets for the treatment of OA,and the selected small molecular drugs can be developed as the key drugs for the treatment of OA.
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BACKGROUND: Pneumatic tourniquet can reduce the amount of bleeding during operation, but simultaneously, pneumatic tourniquet will bring perioperative adverse reactions. It is still controversial whether to use a pneumatic tourniquet during total knee arthroplasty. OBJECTIVE: To evaluate the effect and safety of pneumatic tourniquet in total knee arthroplasty. METHODS: We searched PUBMED, EMBASE, the Cochrane Library, CNKI, Wanfang before July 2017 for randomized controlled trials regarding the effect of pneumatic tourniquet in total knee arthroplasty. Data were extracted and analyzed by using Revman 5.3 software.RESULTS AND CONCLUSION: Totally 18 studies were included. Meta-analysis suggested that the use of pneumatic tourniquet could reduce the total amount of blood loss in the total knee arthroplasty, shorten operation time, but could increase postoperative pain and thrombosis incidence (P < 0.05). There was no statistically significant difference in HSS score and range of motion of the knee at 7 days after operation (P > 0.05). These results confirm that the use of a pneumatic tourniquet in total knee arthroplasty can reduce the total blood loss, shorten the operation time, but have no effect on knee function 7 days after total knee arthroplasty, but will increase postoperative pain and thrombosis incidence.
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Objective To evaluate the therapeutic effects of medial retinaculum tightening under the arthroscope for the adolescent pa -tella dislocation.Methods From January 2010 to July 2016,36 patients(38 knees)of adolescent patellar dislocations were treated by ar-throscope with the medial retinaculum tightening.The patient was evaluated preoperatively and postoperatively,including the J-sign,apprehen-sion test,scores of International Knee Documentation Committee questionnaire(IKDC),Lysholm and Kujala.Results During the followed-up at 3,6 and 12 months after the operation,IKDC scores were improved 35.34%,43.16%and 53.71%respectively compared to the preop-erative data(P<0.01).The Lysholm scores were improved 74.73%,89.89%and 110.9%(P<0.01).The Kujala scores were improved 78.37%,92.62%and 117.8%(P<0.01).All the scores showed a rising trend.There was no significant difference in the scores of pa-tients with acute patellar dislocation group and recurrent patellar dislocation group during the follow -up(P>0.05).The positive rate of J-sign test decreased by 52.63%and the apprehension test decreased by 57.89%(P<0.01)12 months after the operation.IKDC,Kujala and Ly-sholm scores increased obviously after the operation,and the knee joint activity level and overall satisfaction increased significantly.Conclu-sion Arthroscopic treatment with medial retinaculum tightening for adolescent patellar dislocation can result in positive effects,and it is easy to operate and grasp.
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The lentivirus-mediated uPA interference in the proliferation, apoptosis, and secretion of osteoarthritic chondrocytes was examined in this study. Cells were obtained from the cartilage tissues of New Zealand white rabbits. They were cultured with interleukin (IL)-1β (10 ng/mL) for 24 h and then divided into three groups: uPA-siRNA group (cells transfected with uPA-siRNA lentiviruses), blank control group (untreated cells), and negative control group (cells transfected with empty vectors). Western blotting and real-time quantitative reverse transcription-PCR (RT-QPCR) were performed to detect the protein and mRNA expression levels of uPA, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13 and MMP-14 in osteoarthritic chondrocytes. Cell Counting Kit-8, flow cytometry, and colony formation assay were used to examine the proliferation and apoptosis of chondrocytes. The results showed that after uPA-siRNA transfection, the protein and mRNA expression levels of uPA, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13, and MMP-14 were significantly decreased (P<0.05 for MMP-1, MMP-9, MMP-10 and MMP-14, P<0.01 for uPA, MMP-3 and MMP-13). Cell proliferation and colony formation rate were significantly higher and the cell apoptosis rate was significantly lower in uPA-siRNA group than in control groups (P<0.01). The proportion of cells in G0/G1 phase was markedly increased and that in the S phase decreased, and the cell cycle was arrested at the G1/S phase in the control group. In the uPA-siRNA group, the proportion of cells in the S phase was significantly increased, resulting in a different proportion of cells in cell cycle phase (P<0.01). It was suggested that the down-regulation of uPA gene could inhibit the expression of MMPs protein and cell apoptosis, increase the proliferation and colony formation of osteoarthritic chondrocytes.
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The lentivirus-mediated uPA interference in the proliferation, apoptosis, and secretion of osteoarthritic chondrocytes was examined in this study. Cells were obtained from the cartilage tissues of New Zealand white rabbits. They were cultured with interleukin (IL)-1β (10 ng/mL) for 24 h and then divided into three groups: uPA-siRNA group (cells transfected with uPA-siRNA lentiviruses), blank control group (untreated cells), and negative control group (cells transfected with empty vectors). Western blotting and real-time quantitative reverse transcription-PCR (RT-QPCR) were performed to detect the protein and mRNA expression levels of uPA, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13 and MMP-14 in osteoarthritic chondrocytes. Cell Counting Kit-8, flow cytometry, and colony formation assay were used to examine the proliferation and apoptosis of chondrocytes. The results showed that after uPA-siRNA transfection, the protein and mRNA expression levels of uPA, MMP-1, MMP-3, MMP-9, MMP-10, MMP-13, and MMP-14 were significantly decreased (P<0.05 for MMP-1, MMP-9, MMP-10 and MMP-14, P<0.01 for uPA, MMP-3 and MMP-13). Cell proliferation and colony formation rate were significantly higher and the cell apoptosis rate was significantly lower in uPA-siRNA group than in control groups (P<0.01). The proportion of cells in G0/G1 phase was markedly increased and that in the S phase decreased, and the cell cycle was arrested at the G1/S phase in the control group. In the uPA-siRNA group, the proportion of cells in the S phase was significantly increased, resulting in a different proportion of cells in cell cycle phase (P<0.01). It was suggested that the down-regulation of uPA gene could inhibit the expression of MMPs protein and cell apoptosis, increase the proliferation and colony formation of osteoarthritic chondrocytes.
Subject(s)
Animals , Rabbits , Apoptosis , Cell Proliferation , Cells, Cultured , Chondrocytes , Cell Biology , Gene Silencing , Lentivirus , Genetics , Matrix Metalloproteinases , Metabolism , Urokinase-Type Plasminogen Activator , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the methylation Oct4 in orientation induced differentiation in bone marrow mesenchymal stem cells</p><p><b>METHODS</b>Mice BMSCs were isolated and purified from bone marrow by adherent culture,and then identified by morphology and immunocytochemistry.Mouse osteoblastic cells were cultured by bone fragments inoculation,and then identified by alkaline phosphatase(AKP)staining and alizarin red staining.BMSCs were induced to differentiate into osteoblasts in vitro. Indirect immunofluorescence staining and reverse transcription polymerase chain reaction(RT PCR)were used to detect the expressions of Oct4 in BMSCs before and after induction.The methylation status of Oct4 gene in mouse BMSCs was explored by a methylation specific PCR before and after induction</p><p><b>RESULTS</b>The isolated mice BMSCs massively proliferated in vitro and formed cell colones with uniform morphology.Positive expressions of CD29,cKit,and CD44 and negative expression of CD34 were found in the isolated cells.After 10 days[DK]'[DK] induction,both AKP and the alizarin red were positive in cells and osteoblastic cells isolated from mice skull bones.The indirect immunoinfluorescence staining and RT-PCR also showed that the Oct4 expression in the directed differentiation of mouse BMSCs was down-regulated.The CpG island of Otc4 gene promoter in mouse BMSCs became methylated during the induced differentiation.</p><p><b>CONCLUSIONS</b>Mice BMSCs and osteoblasts were successfully cultured in vitro in this studyOct4 may be involved in the maintenance of adult stem cell pluripotency.The down regulated expression of Oct4 gene in mouse BMSCs during the directed differentiation may contribute to the methylation of CpG island in Otc4 gene promoter.</p>
Subject(s)
Animals , Mice , Bone Marrow Cells , Cell Biology , Cell Differentiation , Cells, Cultured , CpG Islands , DNA Methylation , Mesenchymal Stem Cells , Cell Biology , Octamer Transcription Factor-3 , Metabolism , Osteoblasts , Cell Biology , Promoter Regions, GeneticABSTRACT
Quorum sensing systems of pathogens are central regulators for the expression of virulence factors. Increasing evidence implies that targeting the quorum sensing system of many pathogenic bacteria is a promising therapeutic approach to control infections. In this work,we isolated 47 strains of actinomycetes from the mud sample of Jiaozhou Bay. Quorum sensing inhibitory activity was monitored by Chromobacterium violaceum CV026. As a result,the culture broth extract of actinomycetes WA-7 was found to have significant quorum sensing inhibitory activity. This strain was assigned to the genus Streptomyces based on its 16S rDNA sequence. Further investigation revealed that the extract could inhibit the quorum sensing-controlled violacein and proteases production of C. violaceum in a concentration-dependent manner.
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<p><b>OBJECTIVE</b>To describe the prevalence and disability of stroke as well as the stroke-related diseases among elderly in urban and rural regions of Beijing.</p><p><b>METHODS</b>In 2002, three communities were selected from urban, suburb and rural regions from Beijing areas, respectively. Twenty percent of the elderly were randomly selected from three communities. The information about history of stroke, hypertension, heart diseases and diabetes, self-rated health (SRH), activity of daily living (ADL) and instrumental ADL (IADL), smoking and drinking habits, knowledge about cardiovascular diseases prevention were collected.</p><p><b>RESULTS</b>A total of 2487 elderly were interviewed and the prevalence of stroke was 12.9% (321/2481). Eighty-seven of the stroke patients were diagnosed by CT/MRI. 19.9% of stroke patients had experienced 2 or more attacks. The highest prevalence of stroke was in the urban region and the lowest in the rural region (16.9% vs. 8.5%, P for trend < 0.01) while it was higher in males than in females (P < 0.05). The prevalence of stroke tended to increase with age in urban and 34.6% of the stroke patients had recovered completely. The proportions of poor SRH, ADL and IADL dependence, as well as the prevalences of hypertension, heart diseases and diabetes were higher among the elderly with stroke than those without. Although rates of awareness and treatment of hypertension were at the high levels among the elderly with stroke , the control rate was low, especially in the rural region (as low as 4.3%). The level of knowledge on the prevention of cardiovascular diseases, and the rates of smoking and drinking were similar between the elderly with or without stroke.</p><p><b>CONCLUSION</b>The prevalence of stroke had increased dramatically during the past decade in Beijing. The proportion of poor SHR, ADL and IADL dependence, prevalence rates of stroke related diseases were higher among the elderly with stroke than those without. Secondary prevention of stroke among Beijing elderly called for urgent action.</p>