Identification of key pathways and genes involved in microglia inflammation by bioinformatics analysis of transcriptome sequencing / 中华骨科杂志

Objective:To explore the key pathways and genes involved in microglia inflammation through transcriptome sequencing and bioinformatics analysis.Methods:BV2 cells were stimulated by lipopolysaccharide to establish microglia inflammation model. The levels of IL-6 and TNF-α were detected by ELISA and RT-qPCR. The established microglia inflammation model was sequenced by transcriptome sequencing, and the differentially expressed genes were screened by bioinformatics method. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of differentially expressed genes were performed. The protein-protein interaction network of differentially expressed genes was constructed by using string database, and the protein-protein interaction network was visualized by using Cytoscape software. The protein interaction network module was extracted by using MCODE app. The hub gene was extracted by using cytohubba app and was verified through RT-qPCR. We conducted enrichment analysis of hub genes, predicted their targeted miRNAs and interacting drugs.Results:The microglia inflammation model was successfully established and verified by ELISA and RT-qPCR. We screened 434 differentially expressed genes by bioinformatics analysis of transcriptome sequencing results. GO analysis showed that these differentially expressed genes were mainly concentrated in cellular response to cytokine stimulus, inflammatory response, regulation of response to external stimulation. KEGG analysis showed that these differentially expressed genes were mainly concentrated in Chemokine signaling pathway, TNF signaling pathway, IL-17 signaling pathway. We constructed the protein interaction network of these differentially expressed genes, and carried out module analysis and extraction of hub genes. Most of hub genes are located in module 1, and the seed gene of module 1 is S1pr1. Hub genes include S1pr1, Cxcr4, Cx3cl1, Cx3cr1, Cxcl10, Cxcl2, Ccl4, Ccl5, Ccl9, Fpr1. RT-qPCR results showed that compared with the culture medium group, the mRNA expressions of S1pr1, Cxcr4, Cx3cl1 and Cx3cr1 were down-regulated, and the mRNA expressions of Cxcl10, Cxcl2, Ccl4, Ccl5, Ccl9 and Fpr1 were up-regulated in the LPS group. The enrichment analysis of hub genes mainly focused on chemokine-mediated signaling pathway, Class A/1 (Rhodopsin-like receptors), cell chemotaxis and so on. Drugs and miRNAs that may interact with hub genes were predicted. Conclusion:Through transcriptome sequencing and bioinformatics analysis of microglia inflammation model, differentially expressed genes were screened, hub genes and seed genes were extracted, which will help us further understand the molecular mechanism of microglia inflammation and provide potential targets for the treatment of related diseases.

Arthroscopic treatment of tibial eminence avulsion fracture by four-point fixation with suture anchors / 中华创伤骨科杂志

Objective:To explore the clinical effectiveness of arthroscopic treatment of tibial eminence avulsion fracture by four-point fixation with suture anchors.Methods:A retrospective analysis was performed of the 58 patients with tibial eminence avulsion fracture who had been treated by the same group of surgeons using four-point fixation technique with suture anchors under arthroscopy at Department of Sports Medicine, The Affiliated Hospital to Qingdao University from January 2015 to December 2018. They were 33 males and 25 females, with an average age of 18.4 years (from 14 to 32 years). By the modified Meyers-McKeever classification, 15 fractures were type Ⅱ, 19 type Ⅲ and 24 type Ⅳ. Recorded and compared were knee Lysholm scores, International Knee Documentation Committee (IKDC) scores and tibial eminence height between preoperation and one year postoperation; recorded at the last follow-up were range of knee motion and results of Lachman and pivot-shift tests.Results:The 58 patients were followed up for a mean of 20.7 months (from 12 to 33 months). Bony union was achieved in all patients within 12 weeks after operation. In this cohort, the Lysholm score (85.2±4.9) and IKDC score (86.2±4.3) at one year postoperation were significantly higher than the preoperative values (43.2±5.2 and 51.2±4.9), and the post-operative tibial eminence height [(9.1±1.2) mm] was significantly lower than the preoperative value [(12.6±1.2) mm] (all P<0.05). The correlation coefficients between the tibial eminence height and the Lysholm & IKDC scores at one year postoperation were -0.16 and -0.17, respectively. The last follow-up showed a 132°±5° range of knee motion for all patients, a positive result of pivot-shift test (grade Ⅱ) for 3 and a positive result of Lachman test (grade Ⅰ) for 2. Conclusion:Arthroscopic treatment of tibial eminence avulsion fracture by four-point fixation with suture anchors can lead to satisfactory effectiveness, showing advantages of minimal invasion, anatomic reduction, reliable fixation, and little impact on the epiphysis plate.

Depletion of microglia combined with transplantation of bone marrow mesenchymal stem cells for repairing spinal cord injury / 中华骨科杂志

Objective:To study the effect of microglia depletion combined with bone marrow mesenchymal stem cells (BMSC) transplantation for spinal cord injury (SCI) repair.Methods:GFP-BMSCs were cultured, identified and detected for expression levels of growth factors. The effects of BMSCs ondorsal root ganglion (DRG) axon outgrowth were observed by the co-culture of BMSCs with DRGs. Mice were depleted of microglia by administrating the colony stimulating factor 1 receptor (CSF1R) inhibitor PLX3397. The spinal cords of these microglia-depleted mice were subjected to crush injury. BMSCs were transplanted into SCI area after microglia depletion. Mice were randomly divided into control group (SCI+BMSCs) and experimental group (PLX3397+SCI+BMSCs). Mice were sacrificed at corresponding time points after transplantation for observing the survival of transplanted BMSCs and the repair of spinal cord. BMS score was used for evaluation of motor function recovery.Results:BMSCs secreted a large number of neurotrophic factors and promoted the growth of DRG axons when co-cultured with DRGs. Depletion of microglia significantly improved the survival of transplanted BMSCs. Compared with BMSCs transplantation alone, the combined treatments slightly but non-significantly reduced the area of the lesion ( t=2.141, P=0.065). Immunofluorescence staining showed that both BMSC transplantation alone and the combined treatments did not cause the corticospinalaxons across the lesion and into distal spinal cord. BMS scores were (1.20±0.45), (3.20±0.45), (3.80±0.45), (4.20±0.45), and (4.60±0.55) points in control group at 1, 7, 14, 21 and 28 d after injury. The experimental groups were(0.60±0.55), (3.00±0.71), (3.80±0.84), (4.20±0.84), and (4.40±0.89) points, respectively. Conclusion:Depletion of microglia improves the survival of transplanted cells, depletion of microglia combined with BMSC transplantation did not result in a significant reduction in lesion area. At the same time, the damaged CST axons were notregenerated. Thus, combining cell transplantation with axon-promoting strategy may be necessary for SCI repair.

Relationship between fibular head height and varus knee osteoarthritis occurrence and severity / 中华骨科杂志

Objective:To explore the correlation between fibular head height and varus knee osteoarthritis occurrence and severity.Methods:A retrospective analysis was performed on 618 participants (618 knees, 184 males and 434 females, mean age 61.12±10.98 years) who underwent standard weight-bearing full-leg radiographs and were diagnosed as non-knee osteoarthritis or varus knee osteoarthritis from January 2019 to June 2019. Knee osteoarthritis was diagnosed according to Kellgren-Lawrence grading: 0-I grades were diagnosed as non-osteoarthritis, II-IV grades were diagnosed as osteoarthritis. Joint line convergence angle (JLCA), medial proximal tibial angle (MPTA) and hip-knee-ankle angle were measured on X-rays to reflect varus deformity. The fibular head height was defined as the vertical distance from upper edge of fibular head to lateral tibial plateau. Patients were divided into 5 groups according to Kellgren-Lawrence grading. Differences of age, gender, height, weight, body mass index, varus deformity (JLCA, MPTA and hip-knee-ankle angle) between Kellgren-Lawrence 0-IV grades were compared. Ordinal logistic regression was performed to analyze the correlation between fibular head height and Kellgren-Lawrence grades. Pearson's correlation analysis was used for the correlation among fibular head height, JLCA, MPTA and hip-knee-ankle angle, and the main factor of JLCA, MPTA and hip-knee-ankle angle was extracted by factor analysis. Multiple linear regressions were used to analyze the correlation between fibular head height and varus deformity score.Results:There were 68, 66, 97, 98, 289 participants in Kellgren-Lawrence grades 0-IV respectively that was 134 participants were diagnosed as non-osteoarthritis and 484 participants were diagnosed as osteoarthritis. Fibular head height and MPTA showed a decreasing trend ( F=129.076, 24.875; P<0.001) while JLCA and hip-knee-ankle angle showed an increasing trend ( F=414.346, 105.996; P<0.001) with the increase in Kellgren-Lawrence grading. Age, body mass index and fibular head height are influencing factors of Kellgren-Lawrence grading with OR(95%CI) were 1.116(1.093, 1.141), 1.363(1.060, 1.754), 0.617(0.575, 0.662) . Fibular head height was negatively correlated with JLCA and hip-knee-ankle angle ( r=-0.641, -0.478; P<0.001) , respectively, and positively correlated with MPTA ( r=0.320, P<0.001). There were significant correlations between age, fibular head height and the varus deformity score ( β=0.274, -0.457; P<0.001). Conclusion:Fibular head height of patients with varus knee osteoarthritis is lower than that of non-osteoarthritis. In addition to age and body mass index, fibular head height is a risk factor for varus knee osteoarthritis occurrence. The smaller the fibular head height is, the more serious the osteoarthritis severity and varus deformity are.

Research on Risk Control of Upgraded Design and Development Changes of Laser Treatment Equipment after Registration / 中国医疗器械杂志

This paper analyzes the causes and risks of common design and development changes of laser treatment equipment, studies the changes of key components and their corresponding control measures, forms the identification method of design changes of laser treatment equipment, and gives suggestions on how to deal with design and development changes, so as to provide references for inspectors during on-site inspection.

Clinical applications of long non-coding RNA detection in cancers / 中华检验医学杂志

In recent years, the discovery of thousands of long non-coding RNAs ( lncRNAs) has certainly changed human′s view of the complexity of mammalian genomes and transcriptome, as they play an important role in the regulation of transcription, post-transcription as well as epigenetic level, widely involved in various physiological process and diseases.Here this review summarized the lncRNAs associated with cancers, discussed the established methods used to detect and quantify lncRNAs, and evaluated the clinical application of lncRNAs as biomarker.

Influence of chondrocytes of different origin on early differentiation of bone marrow mesenchymal stem cells in co-culture system / 中华老年医学杂志

Objective To investigate the influence of chondrocytes originating from different source on early chondrogenic differentiation of bone marrow mesenchymal stem cells (MSCs) in isolated co-culture system.Methods We applied hanging cell culture system to culture chondrocytes of different origin (osteoarthritis chondrocyte cells,nomal chondrocyte cells,infant chondrocyte cells) and controls.These chondrocytes and MSCs of the same origin were cultured in the common medium in a separated condition,and observed by microscope at 3,6,9,12 day after co culture.Expression levels of aggrecan,collagen type Ⅱ (Col 2),cartilage-specific transcription factor (Sox-9) in MSCs of different origin were determined by Real-time PCR.Results MSCs showed obviously morphological differentiation induced by chondrocytes of different origin at 12 day after coculture as compared with controls.Real-time PCR analysis showed that SOX9 mRNA level was stimulated by 1.7-fold,1.6-fold and 1.2-fold (all P＜0.05) and aggrecan mRNA level was increased by 2.8-fold,2.2-fold and 1.3-fold (all P＜0.05) in infant chondrocytes group,nomal chondrocytes group,osteoarthritis chondrocytes group respectively as compared with controls while COL2 mRNA level had no significant differences among the four groups.Corresponding protein signal level had obvious differences among the four groups,especially in infant chondrocytes as compared with osteoarthritis chondrocytes and nomal chondrocytes.Conclusions Isolated co-culture system may indirectly promote MSCs differentiation to chondrocytes by local micro-environment regulation.Chondrocytes of different origin have different effects on MSCs differentiation,but they could promote MSCs differentiation to chondrocytes.