plac1 gene as a target for diagnosis and treatment of osteoarthritis
A PLAC1, osteoarthritis technology, applied in gene therapy, bone disease, disease diagnosis, etc., can solve problems such as early onset
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Embodiment 1
[0058] Example 1 Expression difference of PLAC1 gene in OA synovial tissue and normal synovial tissue
[0059] The synovial tissues of 8 OA patients were obtained from OA patients undergoing knee arthroplasty or synovectomy in the Department of Orthopedics of Peking Union Medical College Hospital. The cases used were in line with the diagnostic criteria for OA proposed by Altam. 8 cases of normal synovial tissues were from the Department of Orthopedics, Peking Union Medical College Hospital, which were joint synovial tissues of patients undergoing trauma surgery. The synovial fluid (SF) of patients with OA was centrifuged at high speed, and the supernatant was collected and stored at -80°C until use. All clinical samples used in this study were informed to the patients and approved by the Ethics Committee of our hospital.
[0060] 1. Extraction of total RNA from synovial tissue
[0061] The total RNA of the synovial tissue cells of OA patients and normal synovial tissue cel...
Embodiment 2
[0075] Example 2 Large sample verification of differentially expressed genes
[0076] The synovial tissues of 40 OA patients were obtained from OA patients undergoing knee replacement or synovectomy in the Department of Orthopedics of Peking Union Medical College Hospital. The cases used were in line with the diagnostic criteria for OA proposed by Altam. 40 cases of normal synovial tissues were obtained from the Department of Orthopedics of Peking Union Medical College Hospital, which were joint synovial tissues of patients undergoing trauma surgery. The synovial fluid (SF) of patients with OA was centrifuged at high speed, and the supernatant was collected and stored at -80°C until use. All clinical samples used in this study were informed to the patients and approved by the Ethics Committee of our hospital.
[0077] 1. Detection of differential expression of PLAC1 gene at the transcriptional level
[0078] 1.1 Extract synovial tissue total RNA
[0079] Step is with embod...
Embodiment 3
[0109] Example 3 OA fibroblast-like synoviocyte antibody neutralization experiment
[0110] 1. Steps
[0111] 1.1 In vitro culture of synovial tissue cells
[0112] After cleaning the aseptically obtained synovium tissue with PBS, cut it into tissue pieces of about 1mm x 1mm x 1mm repeatedly with sterile surgical scissors, add collagenase II (0.5mg / ml) at 37°C, digest for 2 hours, pass through 200 mesh Filter with gauze, centrifuge to remove supernatant, resuspend cells in DMEM medium, store at 37°C, 5% CO 2 Cultured in a cell culture incubator. After the cells grow into spindles and form sheets, they are subcultured. When the cells were passed to the third passage, FITC-labeled mouse anti-human CD3, CD14, CD19 and PE-labeled mouse anti-human CD11b were added for labeling, and flow cytometry was used for detection and identification. The cells that were negative for the above four markers were fibroblast-like synoviocytes (FLS) and were used in this study.
[0113] 1.3 An...
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