BACKGROUND:As a new treatment method,local injection of medical ozone has anti-inflammatory,analgesic and immunomodulatory effects on injured skeletal muscle tissue,but its mechanism of action is still lacking systematic research.OBJECTIVE:To observe the intervention effect of medical ozone on skeletal muscle crush injury of rabbit models,and to analyze the differential gene expression of injured rabbit tibialis anterior muscle after ozone treatment through RNA-sequencing and tandem mass tag labeled quantitative proteomic technology,in order to partially reveal the molecular mechanism of medical ozone treatment for skeletal muscle injury.METHODS:Eighteen Japanese white rabbits were randomly divided into a blank group,a model group,and an ozone group,with 6 rabbits in each group.Rabbit tibialis anterior muscle compression injury models were established in the latter two groups.After 12 hours of model establishment,2 mL of medical ozone with a mass concentration of 30 μg/mL was injected into the injured muscle of rabbits in the ozone group.Tissue samples were taken 3 days after ozone treatment.Hematoxylin-eosin staining was used to observe the morphological changes of skeletal muscle.RNA sequencing and tandem mass spectrometry tag were used to detect rabbit tibialis anterior muscle tissue.Bioinformatics analysis was used to explore the biological processes of differentially expressed genes and proteins between groups and then to investigate the molecular mechanism of ozone treatment of skeletal muscle injury.RESULTS AND CONCLUSION:After 3 days of ozone treatment,compared with the blank group,the model group showed swelling of cells,infiltration of inflammatory cells,and partial dissolution of muscle fibers.Compared with the model group,cell edema was alleviated and the number of inflammatory cells was reduced in the ozone group.(2)The results of RNA sequencing and bioinformatics analysis showed that compared with the blank group,596 differentially expressed genes were screened in the model group.Compared with the model group,405 differentially expressed genes were screened in the ozone group.There were a total of 194 differentially expressed genes shared among these differentially expressed genes.Proteomic analysis results exhibited that compared with the blank group,the model group contained 138 differentially expressed proteins.Compared with the model group,242 differentially expressed proteins were determined in the ozone group.There were 66 differentially expressed proteins shared between the two comparison groups.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed on differentially expressed genes/proteins,and it was found that they were mainly enriched in the PI3K-Akt signaling pathway,Ras signaling pathway,and MAPK signaling pathway.It is indicated that medical ozone may promote the repair of injured muscle through regulating PI3K/Akt/NF-κB and Ras/MAPK/NF-κB signaling pathway by acting on target genes,such as Syk,FGF16,CSF-1,and MRAS.

BACKGROUND:As a new treatment method,local injection of medical ozone has anti-inflammatory,analgesic and immunomodulatory effects on injured skeletal muscle tissue,but its mechanism of action is still lacking systematic research.OBJECTIVE:To observe the intervention effect of medical ozone on skeletal muscle crush injury of rabbit models,and to analyze the differential gene expression of injured rabbit tibialis anterior muscle after ozone treatment through RNA-sequencing and tandem mass tag labeled quantitative proteomic technology,in order to partially reveal the molecular mechanism of medical ozone treatment for skeletal muscle injury.METHODS:Eighteen Japanese white rabbits were randomly divided into a blank group,a model group,and an ozone group,with 6 rabbits in each group.Rabbit tibialis anterior muscle compression injury models were established in the latter two groups.After 12 hours of model establishment,2 mL of medical ozone with a mass concentration of 30 μg/mL was injected into the injured muscle of rabbits in the ozone group.Tissue samples were taken 3 days after ozone treatment.Hematoxylin-eosin staining was used to observe the morphological changes of skeletal muscle.RNA sequencing and tandem mass spectrometry tag were used to detect rabbit tibialis anterior muscle tissue.Bioinformatics analysis was used to explore the biological processes of differentially expressed genes and proteins between groups and then to investigate the molecular mechanism of ozone treatment of skeletal muscle injury.RESULTS AND CONCLUSION:After 3 days of ozone treatment,compared with the blank group,the model group showed swelling of cells,infiltration of inflammatory cells,and partial dissolution of muscle fibers.Compared with the model group,cell edema was alleviated and the number of inflammatory cells was reduced in the ozone group.(2)The results of RNA sequencing and bioinformatics analysis showed that compared with the blank group,596 differentially expressed genes were screened in the model group.Compared with the model group,405 differentially expressed genes were screened in the ozone group.There were a total of 194 differentially expressed genes shared among these differentially expressed genes.Proteomic analysis results exhibited that compared with the blank group,the model group contained 138 differentially expressed proteins.Compared with the model group,242 differentially expressed proteins were determined in the ozone group.There were 66 differentially expressed proteins shared between the two comparison groups.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed on differentially expressed genes/proteins,and it was found that they were mainly enriched in the PI3K-Akt signaling pathway,Ras signaling pathway,and MAPK signaling pathway.It is indicated that medical ozone may promote the repair of injured muscle through regulating PI3K/Akt/NF-κB and Ras/MAPK/NF-κB signaling pathway by acting on target genes,such as Syk,FGF16,CSF-1,and MRAS.

Kashin-Beck disease (KBD) is an endemic, multiple, symmetrical and deformable osteoarthropathy that is more common in the epiphysis growth plate and articular cartilage of developing children and can cause degeneration, degradation and necrosis of chondrocyte. Clinically, it is divided into four stages based on the extent of the patient's lesion and degree of deformity: early stage, gradeⅠ, gradeⅡ and gradeⅢ. The etiology and pathogenesis of KBD are still unclear, there is no specific drug for this disease. There is also limited study on the effectiveness of various medical interventions. Currently, the treatment of KBD mostly refers to the relevant methods of traditional Chinese and Western medicine for osteoarthritis. This article is based on the existing study, and summarizes and analyzes the Chinese and Western medicine treatment methods for KBD based on the disease staging. It is proposed that both Chinese and Western medicine should be treated under the principles of ladder and individualization, with drug therapy as the main treatment and surgical treatment as the auxiliary treatment, so as to minimize patient pain, protect joints and prevent injuries, and provide reference for clinical treatment of KBD.

Objective To observe the effects of intra-articular injection of sinomenine on the expressions of vascular endothelial growth factor (VEGF) and nerve growth factor (NGF) in articular cartilage and synovium of rabbit knee osteoarthritis models; To discuss its mechanism of action. Methods Totally 40 New Zealand white rabbits were randomly divided into 4 groups: control group, model group, hyaluronate group and sinomenine group. Model group, hyaluronate group and sinomenine group established knee osteoarthritis model by using intra-articular injection of papain enzyme and were treated with saline, sodium hyaluronate and sinomenine respectively for 30 d, except for the control group. After treatment, method of immunohistochemistry was used to detect the expressions of VEGF and NGF in rabbit articular cartilage and synovium. Results Compared with the control group, the expressions of VEGF and NGF in articular cartilage and synovium of model group significantly increased (P<0.01); compared with the model group, the expressions of VEGF and NGF in articular cartilage and synovium of sinomenine group significantly decreased (P<0.05, P<0.01); the expressions of VEGF and NGF in articular cartilage and synovium in sinomenine group were lower than hyaluronate group (P<0.05, P<0.01). Conclusion Intra-articular injection of sinomenine can down-regulate the expressions of VEGF and NGF in articular cartilage and synovium of rabbit knee osteoarthritis.

Objective To observe the effects of intra-articular injection of sinomenine on morphology of synovium and cartilage as well as contents of MMP-13 and cartilage oligomeric matrix protein(COMP) in serum and synovial fluid of rabbit knee osteoarthritis model.Methods A total of 38 New Zealand white rabbits were randomly divided into 4 groups: control group, model group, hyaluronate group and sinomenine group. Model group, hyaluronate group and sinomenine group established knee osteoarthritis model by intra-articular injection of papain enzyme, and were treated with saline, sodium hyaluronate and sinomenine respectively except for control group. 5 weeks after treatment, all rabbits were sacrificed for HE staining and histological grading on cartilago articularis and synovium, and ELISA method was used to detect the contents of MMP-13 and COMP in rabbit serum and synovial fluid.Results Mankin's scores of articular cartilage and histological scores of synovium in model group were significantly higher than those of control group (P<0.01), and were markedly lower in sinomenine group than those of model group (P<0.01); the contents of MMP-13 and COMP in serum and synovial fluid of model group were significantly raised compared with control group (P<0.01), and were reduced obviously in sinomenine group compared with model group (P<0.05,P<0.01).Conclusion Intra-articular injection of sinomenine can reduce the levels of MMP-13 and COMP in serum and synovial fluid of rabbit knee osteoarthritis model, and improve synovial inflammation, as well as delay the degradation of articular cartilage.