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Wfs1 as a novel molecular marker

A WFS1, a pair of technology, applied in the field of diagnosis and treatment of WFS1 gene in rheumatoid arthritis and osteoarthritis, can solve the problems of difficult identification of clinical features and difficult diagnosis

Active Publication Date: 2021-03-02
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In primary care, the diagnosis of early rheumatoid arthritis is as difficult as other musculoskeletal problems: clinical features are poorly discernible, inflammatory markers such as erythrocyte sedimentation rate and C-reactive protein are not characteristic, and specific markers such as rheumatoid factor ( Seronegative in 31% of patients) and anti-cyclic citrullinated peptide antibodies (anti-CCP, seronegative in 33% of patients) were also commonly seronegative
There are no molecular markers for the differential diagnosis of rheumatoid arthritis and osteoarthritis, and the applicant carried out this study under such a background

Method used

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  • Wfs1 as a novel molecular marker
  • Wfs1 as a novel molecular marker
  • Wfs1 as a novel molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Screening of differentially expressed genes in osteoarthritis synovial tissue, rheumatoid arthritis synovial tissue and normal synovial tissue

[0056] The synovial tissue of 5 patients with rheumatoid arthritis came from patients with rheumatoid arthritis who underwent knee replacement or synovectomy in the orthopedics department of the hospital. The diagnosis of all cases met the classification criteria of rheumatoid arthritis revised by the American Rheumatology Association in 1987 , including 3 females and 2 males, with an average age of (55±8) years and an average disease duration of (12±7) years. The synovial tissues of 5 patients with osteoarthritis were obtained from osteoarthritis patients undergoing knee joint replacement or synovectomy in the Department of Orthopedics of the hospital. Age (63±5) years old, mean disease duration (12±8) years. 6 cases of normal synovial tissues came from joint synovial tissues of patients undergoing trauma surgery. ...

Embodiment 2

[0103] Example 2 Large sample verification of differentially expressed genes

[0104] Based on the previous high-throughput sequencing results and according to the size of the P value, we selected the WFS1 gene for verification. The down-regulation was even greater in patients with osteoarthritis.

[0105] 1. Detection of differential expression of WFS1 gene at the transcriptional level

[0106] 1. According to the method of Example 1, 100 cases of rheumatoid arthritis synovial tissue, 100 cases of osteoarthritis synovial tissue, and 100 cases of normal synovial tissue were collected.

[0107] 2. Carry out the extraction and quality detection of RNA according to the manner of the examples

[0108] 3. Reverse transcription

[0109] 1 μg of total RNA was reverse-transcribed to synthesize cDNA using reverse transcription buffer. Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, add the following components to the PCR tube respectively: DEPC water, ...

Embodiment 3

[0130] Example 3 WFS1 gene expression plasmid construction

[0131] 1. Construction of WFS1 gene expression vector

[0132] Amplification primers were designed according to the coding sequence of the WFS1 gene (as shown in SEQ ID NO.5). The coding sequence of the full-length WFS1 gene was amplified from the cDNA library of adult fetal brain (clontech company, catalog number: 638831), and the above cDNA sequence was double-digested with restriction endonucleases BamHI and XhoI and inserted into the The obtained recombinant vector pcDNA3.1-WFS1 was connected to the eukaryotic cell expression vector pcDNA3.1 double-digested with enzymes BamHI and XhoI for subsequent experiments.

[0133] 2. In vitro culture of synovial tissue cells

[0134] After the aseptically obtained synovial tissues of rheumatoid arthritis and osteoarthritis were washed with PBS, they were repeatedly cut into tissue pieces of about 1mm x 1mm x 1mm with sterile surgical scissors, and collagenase II (0.5mg / ...

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Abstract

The invention discloses WFS1 as a target for diagnosis and treatment of rheumatoid arthritis and / or osteoarthritis. Accordingly, WFS1 can be used to develop products for diagnosing rheumatoid arthritis and / or osteoarthritis, and develop drugs for treating rheumatoid arthritis and / or osteoarthritis. The research results of the present invention provide a theoretical basis for clinicians to formulate personalized treatment plans, and can provide new drug targets for the development of colon adenocarcinoma drugs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of WFS1 gene in the diagnosis and treatment of rheumatoid arthritis and osteoarthritis. Background technique [0002] Rheumatoid arthritis is an autoimmune disease characterized by polyarticular involvement that can progress to joint destruction and deformity, often involving peripheral joints. The etiology is unclear, but extraarticular organ involvement such as pulmonary interstitial disease and Sjogren's syndrome are also common. Appropriate early treatment can improve joint symptoms and function, reduce mortality, and reduce complications. [0003] A 2010 systematic review found that the prevalence of rheumatoid arthritis in North America and Northern Europe was estimated to range from 0.5% to 1.1%. The incidence in developing countries is relatively lower (0.1% to 0.5%). Rheumatoid arthritis is more common in women (1:3 incidence of men and women). Onset can occur at...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883C12Q2600/158G01N33/6893G01N2800/102G01N2800/105
Inventor 肖枫李曙光孙耀兰常鹏
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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