BACKGROUND: Double-stranded RNA-specific adenosine deaminase 1 (ADAR1) binds to double-stranded RNA and catalyzes the deamination of adenosine (A) to inosine (I). The functional mechanism of ADAR1 in non-small cell lung cancer (NSCLC) remains incompletely understood. This study aimed to investigate the prognostic significance of ADAR1 in NSCLC and to elucidate its potential role in regulating tumor cell proliferation and migration. METHODS: Data from The Cancer Genome Atlas (TCGA) and cBioPortal were analyzed to assess the correlation between high ADAR1 expression and clinicopathological features as well as prognosis in lung cancer. We performed Western blot (WB), cell proliferation assays, Transwell invasion/migration assays, and nude mouse xenograft modeling to examine the phenotypic changes and molecular mechanisms induced by ADAR1 knockdown. Furthermore, the ADAR1 p150 overexpression model was utilized to validate the proposed mechanism. RESULTS: ADAR1 expression was significantly elevated in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) tissues compared with adjacent non-tumor tissues (LUAD: P=3.70×10-15, LUSC: P=0.016). High ADAR1 expression was associated with poor prognosis (LUAD: P=2.03×10-2, LUSC: P=2.81×10-2) and distant metastasis (P=0.003). Gene Set Enrichment Analysis (GSEA) indicated that elevated ADAR1 was associated with mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway activation, matrix metalloproteinase-9 (MMP-9) expression, and cell adhesion. ADAR1 and MMP-9 levels showed a strongly positive correlation (P=6.45×10-34) in 10 lung cancer cell lines, highest in H1581. Knockdown of ADAR1 in H1581 cells induced a rounded cellular morphology with reduced pseudopodia. Concomitantly, it suppressed cell proliferation, invasion, migration, and in vivo tumorigenesis. It also suppressed ERK phosphorylation and downregulated cellular Finkel-Biskis-Jinkins murine osteosarcoma viral oncogene homolog (c-FOS), MMP-9, N-cadherin, and Vimentin. Conversely, ADAR1 p150 overexpression in PC9 cells enhanced ERK phosphorylation and increased c-FOS and MMP-9 expression. CONCLUSIONS High ADAR1 expression is closely associated with poor prognosis and distant metastasis in NSCLC patients. Mechanistically, ADAR1 may promote proliferation, invasion, migration, and tumorigenesis in lung cancer cells via the ERK/c-FOS/MMP-9 axis.
Humans ; Lung Neoplasms/physiopathology* ; Adenosine Deaminase/genetics* ; Matrix Metalloproteinase 9/genetics* ; Cell Proliferation ; Carcinoma, Non-Small-Cell Lung/physiopathology* ; Cell Movement ; Animals ; Mice ; RNA-Binding Proteins/genetics* ; Female ; Male ; Cell Line, Tumor ; Proto-Oncogene Proteins c-fos/genetics* ; Middle Aged ; MAP Kinase Signaling System ; Gene Expression Regulation, Neoplastic ; Mice, Nude ; Extracellular Signal-Regulated MAP Kinases/genetics*

BACKGROUND:Exosomes have been confirmed to be closely related to cartilage degeneration in osteoarthritis.However,the role and mechanism of exosome-derived genes in cartilage degeneration of osteoarthritis have not been fully elucidated.OBJECTIVE:Bioinformatics analyses were used to screen the genes related to cartilage degeneration in the synovial exosomes of patients with osteoarthritis,and to determine their biological functions and signaling pathways in order to provide new therapeutic targets for delaying cartilage degeneration in osteoarthritis.METHODS:Firstly,osteoarthritis-related exosome dataset GSE185059 and cartilage degeneration dataset GSE114007 were downloaded from Gene Expression Omnibus(GEO)database to screen exosome-derived cartilage degeneration related genes.GO functional and KEGG pathway enrichment analyses were performed based on the screened exosome-derived cartilage degeneration related genes.Protein-protein interaction network was drawn and Ingenuity Pathway Analysis(IPA)was conducted to screen and obtain key exosome-derived cartilage degeneration-related genes.Finally,qRT-PCR was used to verify the expression of key genes in osteoarthritis cartilage tissue and interleukin-1β stimulated chondrocyte models.RESULTS AND CONCLUSION:(1)There were 831 differentially expressed genes in the GSE185059 dataset and 5 323 differentially expressed genes in the GSE114007 dataset.A total of 94 exosome-derived cartilage degeneration related genes were screened after the intersection of these differentially expressed genes,of which 51 genes were down-regulated and 43 genes were up-regulated.(2)GO functional enrichment analysis showed that the up-regulated genes were mainly involved in the positive regulation of cell-cell adhesion,the positive regulation of T cell activation,and chronic inflammatory response,while the down-regulated genes were mainly involved in biological processes such as cell aggregation,cartilage differentiation and development,and skeletal system morphogenesis.(3)KEGG pathway enrichment analysis showed that exosome-derived cartilage degeneration-related genes were mainly involved in tryptophan enrichment metabolism,vitamin B6 metabolism,and leukocyte transendothelial migration.(4)The constructed protein-protein interaction network confirmed the existence of multiple interaction relationships among exosome-derived cartilage degeneration-related genes.Combined with five algorithms in CytoHubba software,four key exosome-derived cartilage degeneration-related genes were further screened,namely THY1,CYP1A1,NFKB2,and COL6A3.(5)The results of qRT-PCR showed that compared with normal cartilage,the expressions of THY1 and COL6A3 in osteoarthritic cartilage were increased,while the expression of CYP1A1 and NFKB2 was decreased.Similarly,compared with the unstimulated group,the expression of THY1 and COL6A3 in the interleukin-1β induced chondrocytes was upregulated,while the expression of CYP1A1 and NFKB2 was downregulated.(6)These results indicate that THY1,CYP1A1,NFKB2,and COL6A3 are genes related to cartilage degeneration in the exosomes of synovial fluid of patients with osteoarthritis,and may participate in the pathogenesis of osteoarthritis by regulating biological processes such as protein tyrosine kinase activity and lipid metabolism,as well as nuclear factor-κB signaling pathway and focal adhesion signaling pathway.However,the specific regulatory roles and molecular mechanisms of these key genes in cartilage degeneration need to be further verified by experiments.

BACKGROUND:Exosomes have been confirmed to be closely related to cartilage degeneration in osteoarthritis.However,the role and mechanism of exosome-derived genes in cartilage degeneration of osteoarthritis have not been fully elucidated.OBJECTIVE:Bioinformatics analyses were used to screen the genes related to cartilage degeneration in the synovial exosomes of patients with osteoarthritis,and to determine their biological functions and signaling pathways in order to provide new therapeutic targets for delaying cartilage degeneration in osteoarthritis.METHODS:Firstly,osteoarthritis-related exosome dataset GSE185059 and cartilage degeneration dataset GSE114007 were downloaded from Gene Expression Omnibus(GEO)database to screen exosome-derived cartilage degeneration related genes.GO functional and KEGG pathway enrichment analyses were performed based on the screened exosome-derived cartilage degeneration related genes.Protein-protein interaction network was drawn and Ingenuity Pathway Analysis(IPA)was conducted to screen and obtain key exosome-derived cartilage degeneration-related genes.Finally,qRT-PCR was used to verify the expression of key genes in osteoarthritis cartilage tissue and interleukin-1β stimulated chondrocyte models.RESULTS AND CONCLUSION:(1)There were 831 differentially expressed genes in the GSE185059 dataset and 5 323 differentially expressed genes in the GSE114007 dataset.A total of 94 exosome-derived cartilage degeneration related genes were screened after the intersection of these differentially expressed genes,of which 51 genes were down-regulated and 43 genes were up-regulated.(2)GO functional enrichment analysis showed that the up-regulated genes were mainly involved in the positive regulation of cell-cell adhesion,the positive regulation of T cell activation,and chronic inflammatory response,while the down-regulated genes were mainly involved in biological processes such as cell aggregation,cartilage differentiation and development,and skeletal system morphogenesis.(3)KEGG pathway enrichment analysis showed that exosome-derived cartilage degeneration-related genes were mainly involved in tryptophan enrichment metabolism,vitamin B6 metabolism,and leukocyte transendothelial migration.(4)The constructed protein-protein interaction network confirmed the existence of multiple interaction relationships among exosome-derived cartilage degeneration-related genes.Combined with five algorithms in CytoHubba software,four key exosome-derived cartilage degeneration-related genes were further screened,namely THY1,CYP1A1,NFKB2,and COL6A3.(5)The results of qRT-PCR showed that compared with normal cartilage,the expressions of THY1 and COL6A3 in osteoarthritic cartilage were increased,while the expression of CYP1A1 and NFKB2 was decreased.Similarly,compared with the unstimulated group,the expression of THY1 and COL6A3 in the interleukin-1β induced chondrocytes was upregulated,while the expression of CYP1A1 and NFKB2 was downregulated.(6)These results indicate that THY1,CYP1A1,NFKB2,and COL6A3 are genes related to cartilage degeneration in the exosomes of synovial fluid of patients with osteoarthritis,and may participate in the pathogenesis of osteoarthritis by regulating biological processes such as protein tyrosine kinase activity and lipid metabolism,as well as nuclear factor-κB signaling pathway and focal adhesion signaling pathway.However,the specific regulatory roles and molecular mechanisms of these key genes in cartilage degeneration need to be further verified by experiments.

【Objective】 To analyze the effect of different range and location of foramen formation on the biomechanics of lumbar spine by three-dimensional finite element analysis （D-FEA）. 【Methods】 A complete model of the lumbar spine （L5）， M0， was developed using the finite element method， and the models M1， M2， M3， M4 and M5 were obtained by sequentially simulating the apical， medial 1/4， 2/4， 3/4 and 4/4 graded resections of the left superior articular process of L5 under a lateral posterior approach with full spinal endoscopy. The displacements were recorded in six conditions： forward flexion， back extension， left and right lateral bending， and left and right lateral rotation. The results were compared between the resected models and the unresected group M0. 【Results】 The three-dimensional finite model of the L4-L5 segment developed in this experiment was valid. Compared with the unresected group M0， the differences in ROM were statistically significant for M1 under forward flexion load （all P<0.05）， M2 under forward flexion and back extension load （all P<0.05）， M3 and M4 under forward flexion， back extension and left and right lateral bending load （all P<0.05）. The differences were statistically significant for M3 and M4 under anterior flexion， posterior extension， left and right lateral flexion， and right rotation loads （all P<0.05）； and for M5 under anterior flexion， posterior extension， left and right lateral flexion and right rotation loads （all P<0.05）. Compared with M0 in the unresected group， the differences were statistically significant for M1 under anterior flexion loads （all P<0.05）， M2 under anterior flexion and left and right rotation loads （all P<0.05）. The differences were statistically significant for M3， M4 and M5 in forward flexion and extension， left and right lateral flexion， and left and right rotational loading （all P<0.05）. 【Conclusion】 In the process of foramen formation， removal of the tip or the medial quarter of the unilateral single segment of the upper articular process of the lumbar spine will affect the stability of the lumbar spine， and increase the maximum value of the stress of the intervertebral disc during the activities of the lumbar spine. Removal of one half or more will significantly damage the biomechanics of the lumbar spine. In order to avoid damaging the normal biomechanics of the lumbar spine， the upper articular process should be protected as much as possible during the whole spinal endoscopic foraminal reconstruction.

【Objective】 To observe the short-term clinical effect of selective nerve block combined with percutaneous posterior endoscopic cervical discectomy （PPECD） in the treatment of cervical spondylotic radiculopathy. 【Methods】 We reviewed 22 patients who received selective nerve block combined with percutaneous posterior endoscopic cervical discectomy （PPECD） for cervical spondylotic radiculopathy from June 2018 to January 2020. We recorded Japanese Orthopaedic Association （JOA） score for treatment evaluation， visual analogue scale （VAS）， the neck disability index score （NDI） preoperative 1 day， postoperative 1 day， 1 month， 3 months， 6 months and 1 year. All data were imported into SPSS26.0 software for analysis and processing. Quantitative data are expressed as mean ± standard deviation. The scores of neck VAS， arm VAS， JOA and NDI were compared at different time points by repeated measurement analysis of variance. Paired t-test was used to compare each time point after operation and the first day before operation. P<0.05 was considered statistically significant， and the modified MacNab standard was used to evaluate the clinical effect at the last follow-up. 【Results】 All operations were successfully completed under ultrasound-guided selective nerve block combined with endoscopic operation. The average operation time was 125.6 minutes. The intraoperative blood loss was 2-100 mL and the average blood loss was 19.1 mL. All patients were followed up for 15-33 months， with an average follow-up of 24.1 months. No patients had spinal cord， nerve root and vascular injury， dural tear or other complications. Compared with the preoperative VAS score， the VAS score of neck and upper arm decreased significantly （P<0.05）， while the JOA score increased significantly （P<0.05）， and the preoperative NDI score decreased significantly （P<0.05）. At the last follow-up， the modified Macnab criteria showed there were 15 excellent cases， 5 good cases， 2 medium cases and 0 poor case. The excellence rate was 91%. Postoperative magnetic resonance imaging and 3D computed tomography reconstruction of the cervical spine showed that the disc had been fully removed and the nerve root compression at the surgical segment was relieved. 【Conclusion】 Ultrasound-guided selective nerve block combined with percutaneous posterior endoscopic cervical discectomy is a safe and effective minimally invasive surgical method for the treatment of cervical spondylotic radiculopathy with reliable short-term efficacy.