Reagents for detecting and targeting rp11-754b17.1 and their application in arthritis
A technology for osteoarthritis and reagents, which is applied in the field of reagents for detecting and targeting the biomarker RP11-754B17.1, which can solve the problems of no effective treatment, the influence of patients' families and society, and the inability to cure osteoarthritis.
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Embodiment 1
[0054] Example 1 QPCR sequencing to verify the expression of RP11-754B17.1 gene
[0055] 1. Sample collection
[0056] A total of 78 patients undergoing knee replacement surgery were selected, including 48 OA patients and 30 normal controls. Blood and tissue samples were collected from the patients. The age and gender of the OA group and the control group were not statistically significant.
[0057] OA group: Inclusion criteria: 1) According to the diagnostic criteria of OA, patients with osteoarthritis were diagnosed and underwent knee replacement surgery; 2) First replacement. Exclusion criteria: 1) Complicated with other inflammatory arthritis or autoimmune diseases, including rheumatoid arthritis, gout, systemic lupus erythematosus, etc.; 2) History of severe knee joint trauma; 3) Steroids in the past 3 months History of drug injection or non-steroid drug use; 4) Patients with severe liver and kidney disease and cardiovascular disease.
[0058] Control group: Inclusion criteria: ...
Embodiment 2
[0068] Example 2 Silencing of RP11-754B17.1 gene and its effect on cells
[0069] 1. Cell acquisition and culture
[0070] Wash the cartilage tissue of patients with OA 3 times with 1% bi-antibody PBS, and cut the tissue to 1 mm with sterile ophthalmic scissors 3 Put the obtained cartilage fragments into a l0cm petri dish, add l0ml fresh 0.25% trypsin, 37℃, 5% CO 2 Digest in an incubator for 30 minutes to remove fibroblasts that may adhere. Aspirate the trypsin digestion solution, add 10ml of DMEM culture solution containing 10% FBS, pipetting and mixing to terminate the digestion, and wash twice with 10ml of PBS containing double antibody. Add 10ml of 0.2% type II collagenase to the petri dish, place it at 37℃, 5% CO 2 Digest in the incubator for 4h. The free chondrocyte suspension containing collagenase solution was sucked out with a pipette, filtered through a 70 μm 200 mesh cell sieve, and the resulting filtrate was transferred into a centrifuge tube and centrifuged at 1000 r / ...
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