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Application of SPRR1A gene in preparation of osteoarthritis diagnosis product

An osteoarthritis and gene technology, applied in the application field of SPRR1A gene in the preparation of osteoarthritis diagnostic products

Active Publication Date: 2016-08-24
肖刻
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although various drug therapies have been used to treat the disease, they are ineffective for long-term control and prevention

Method used

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  • Application of SPRR1A gene in preparation of osteoarthritis diagnosis product
  • Application of SPRR1A gene in preparation of osteoarthritis diagnosis product
  • Application of SPRR1A gene in preparation of osteoarthritis diagnosis product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 High-throughput sequencing screening of differentially expressed genes

[0070] 1. Sampling

[0071] From October 2012 to December 2015, patients with osteoarthritis who were treated in the Department of Orthopedics of Peking Union Medical College Hospital were collected. A total of 10 cases were collected as the case group. A total of 4 cases were collected from patients with other diseases who were hospitalized in the Department of Orthopedics during the same period as controls. The synovial fluid samples of all subjects were obtained, numbered and stored in a -80°C low-temperature refrigerator.

[0072] The patients in the case group were all in line with the diagnostic criteria of knee OA and underwent knee arthroplasty; the control group were patients with meniscus injury and cruciate ligament injury who underwent arthroscopic surgery. According to the 1995 American Rheumatology Association diagnostic criteria for osteoarthritis, a patient with severe os...

Embodiment 2

[0094] Example 2 RT-PCR verification of SPRR1A gene expression in synovial fluid of patients with osteoarthritis and controls

[0095] 1. Materials

[0096] The cartilage tissues of 42 cases of osteoarthritis patients and 8 cases of control cartilage tissues were selected and grouped and numbered. The patients in the case group were all in line with the diagnostic criteria of knee OA and underwent knee arthroplasty; the control group were patients with meniscus injury and cruciate ligament injury who underwent arthroscopic surgery.

[0097] 2. Method

[0098] 2.1 Carry out total RNA extraction to synovial fluid, with the extraction method of embodiment 1.

[0099] 2.2 Synthesis of cDNA by reverse transcription

[0100] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0101] Using a reverse transcri...

Embodiment 3

[0119] Example 3 Gene chip verification of SPRR1A gene expression in synovial fluid of patients with osteoarthritis and controls

[0120] 1, the acquisition of material, with embodiment 2.

[0121] 2, the extraction of total RNA, with the method for embodiment 1.

[0122] 3. Gene chip verification

[0123] After the total RNA was linearized and amplified, the cy3-UTP labeled and fluorescently labeled cRNAs were purified using the RNEASY Mini Kit, and the labeled cRNAs were fragmented with Amhion’s RNA Fragmentation Reagents. Human whole gene expression profile chips (4x 44K genes) from Agilent Company of the United States were used, hybridized in a chip hybridization oven at 65°C for 17 hours, then eluted, stained, and finally scanned with an Agilent DNA MicroarrayScanner scanner.

[0124] After the hybridized chip reads the data points by the chip scanner, the data is imported into the analysis software, and the genes whose absolute value of the natural logarithm of the rat...

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Abstract

The invention discloses application of an SPRR1A gene in preparation of an osteoarthritis diagnosis product, discloses the SPRR1A gene related to osteoarthritis and further proves down-regulation of the SPRR1A gene or expression proteins of the gene and fragments, analogs and derivatives of the gene in osteoarthritis tissue. By detecting osteoarthritis with the gene, it is possible to quickly and effectively provide early detection and also to provide a treatment target and an important basis for clinical application such as gene therapy and drug therapy.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of SPRR1A gene in the preparation of osteoarthritis diagnostic products. Background technique [0002] Osteoarthritis (OA) is a chronic disease in which the articular cartilage, which coats the ends of bones and forms the articular surfaces of joints, gradually deteriorates over time. A number of factors have been reported to predispose patients to osteoarthritis, including genetic predisposition, obesity, accident or sports trauma, surgery, medications, and strenuous demands. Osteoarthritis is thought to begin with damage to the articular cartilage. The two most common injuries to joints are sports-related injuries and long-term "repeated use" joint injuries. The joints most often affected by osteoarthritis are the knees, hips, and hands. In most cases, osteoarthritis in the knees and hips is more disabling than hand osteoarthritis due to the essential weight-bearin...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68
CPCC12Q1/6837C12Q1/6841C12Q1/686C12Q1/6883G01N33/68
Inventor 肖刻翁习生吴志宏邱贵兴朱乾坤
Owner 肖刻
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