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Application of KHDC1L gene serving as disease diagnosis marker

A technology of gene and gene expression, applied in the field of diagnosis and treatment of osteoarthritis of human KHDC1L gene, can solve problems such as disability

Active Publication Date: 2016-01-20
GUAN BOJIAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as the disease develops and serious consequences lead to loss of joint function, it is likely to be disabled

Method used

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  • Application of KHDC1L gene serving as disease diagnosis marker
  • Application of KHDC1L gene serving as disease diagnosis marker
  • Application of KHDC1L gene serving as disease diagnosis marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Differences in the expression of KHDC1L gene in embodiment 1OA synovial tissue and normal synovial tissue

[0060] 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.

[0061] 1. Extraction of total RNA from synovial tissue

[0062] After collecting the samples, freeze them in liquid nitrogen. After taking th...

Embodiment 2

[0082] Example 2 Large sample verification of differentially expressed genes

[0083] 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.

[0084] 1. Detection of differential expression of KHDC1L gene at the transcriptional level

[0085] 1.1 Extract synovial tissue total RNA

[0086] Step is with embo...

Embodiment 3

[0104] Example 3 OA fibroblast-like synoviocyte antibody neutralization experiment

[0105] 1. Steps

[0106] 1.1 In vitro culture of synovial tissue cells

[0107] After the aseptically obtained synovium tissue was washed with PBS, it was repeatedly cut into tissue pieces of about 1mmx1mmx1mm with sterile surgical scissors, added with collagenase II (0.5mg / ml) at 37°C, digested for 2 hours, passed through a 200-mesh gauze After filtration and centrifugation to remove the supernatant, the cells were resuspended in DMEM medium and placed 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. Cells negative for the above four markers were fibroblast-like synoviocytes (FLS) used in this ...

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Abstract

The invention discloses that a KHDC1L gene and KHDC1L protein can be served as molecular markers of osteoarthritis diagnosis. The invention uses tests to prove that: compared with a normal person, the expression quantity of the KHDC1L gene and the KHDC1L protein in the synovial tissues of a patient with osteoarthritis is high, and the difference has statistical significance. The invention also discloses that the KHDC1L gene and the KHDC1L protein can be used for preparing the medicine for curing osteoarthritis, and a cell proliferation test proves that by disturbing the KHDC1L gene or inhibiting the activity of the KHDC1L protein, synoviocyte proliferation can be inhibited. The research result of the invention provides a new bone and arthrosis clinical diagnosis method, and simultaneously provides a medicine new target for curing bone and arthrosis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of human KHDC1L gene in the diagnosis and treatment of osteoarthritis. Background technique [0002] China has gradually stepped into an aging society, and the degree of impact of OA on human health and the resulting medical expenses are increasing at an alarming rate year by year. According to data, 50% of the population over the age of 60 have the performance of OA on X-ray films, of which 35% to 50% have clinical symptoms, and more than 80% of the population over the age of 75 have symptoms of OA. A very dire proportion. Now, OA has become the second major disease after heart disease which incapacitates patients and brings heavy economic burden to families and society, which has attracted widespread attention from professionals at home and abroad. [0003] At present, the definition of OA in the guidelines for the diagnosis and treatment of osteoarthritis is as f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11G01N33/68A61K45/00A61K48/00A61K31/7088A61P19/02
CPCA61K31/7088A61K45/00C12Q1/6883C12Q2600/158G01N33/6893G01N2800/105
Inventor 杨承刚杜玉梅
Owner GUAN BOJIAN BIOTECH CO LTD
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