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WFS1 (Wolfram 1) as new molecular marker

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

Active Publication Date: 2018-09-04
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 (Wolfram 1) as new molecular marker
  • WFS1 (Wolfram 1) as new molecular marker
  • WFS1 (Wolfram 1) as new 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 tissues of 5 patients with rheumatoid arthritis came from patients with rheumatoid arthritis who underwent knee joint replacement or synovectomy in the orthopedic department of the hospital. The diagnosis of all cases met the classification criteria for rheumatoid arthritis revised by the American Rheumatoid Association in 1987 Among them, there were 3 females and 2 males, with an average age of (55±8) years and an average duration of (12±7) years. The synovial tissues of 5 patients with osteoarthritis came from osteoarthritis patients who underwent knee replacement or synovectomy in the orthopedics department of the hospital. The cases used met the diagnostic criteria for OA proposed by Altam, including 2 females and 3 males. The average Age (63±5) years old, average course of disease (12±8) years. ...

Embodiment 2

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

[0104] Based on the results of previous high-throughput sequencing and according to the value of P value, we selected the WFS1 gene for verification. Its expression is down-regulated in patients with rheumatoid arthritis and osteoarthritis, and compared with patients with bone and joints, it is The downward adjustment is greater in patients with arthritis.

[0105] 1. Detect the differential expression of WFS1 gene at the transcription level

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

[0107] 2. Perform RNA extraction and quality inspection according to the embodiment

[0108] 3. Reverse transcription

[0109] Use reverse transcription buffer to reverse transcription of 1μg total RNA to synthesize cDNA. Using 25μl reaction system, each sample takes 1μg total R...

Embodiment 3

[0130] Example 3 Construction of WFS1 gene expression plasmid

[0131] 1. Construction of WFS1 gene expression vector

[0132] According to the coding sequence of WFS1 gene (as shown in SEQ ID NO. 5), the amplification primers are designed. The full-length WFS1 gene coding sequence was amplified from the adult fetal brain cDNA library (clontech company, catalog number: 638831). The above cDNA sequence was double-enzyme digested with restriction enzymes BamHI and XhoI and inserted into the restriction endonuclease. In the eukaryotic expression vector pcDNA3.1 digested with enzymes BamHI and XhoI, the recombinant vector pcDNA3.1-WFS1 obtained by ligation was used in subsequent experiments.

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

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

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Abstract

The invention discloses WFS1 (Wolfram 1) as a treatment target of rheumatoid arthritis and / or osteoarthritis. Hereby, the WFS1 can be used for developing products for diagnosing the rheumatoid arthritis and / or the osteoarthritis and developing drugs for treating the rheumatoid arthritis and / or the osteoarthritis. A research result of the invention provides fundamental basis for a clinicist to makean individual treatment program and is capable of providing a new drug target for development of colon adenocarcinoma drugs.

Description

Technical field [0001] The present 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 multi-joint involvement, which can progress to joint destruction and deformity, often involving peripheral joints. The etiology is unclear, and involvement of extra-articular organs such as interstitial lung 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 incidence of rheumatoid arthritis in North America and Northern Europe is estimated to be 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 (male and female incidence rate 1:3). The disease can occur at...

Claims

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Application Information

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Patent Type & Authority Applications(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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