Marker for detecting osteoarthritis and application thereof

A technology for osteoarthritis and markers, applied in the field of biomedical testing, can solve problems such as lack of

Inactive Publication Date: 2018-11-30
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, due to the lack of fundamental understanding of the etiology of OA, the medical community still lacks effective

Method used

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  • Marker for detecting osteoarthritis and application thereof
  • Marker for detecting osteoarthritis and application thereof
  • Marker for detecting osteoarthritis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The collection of embodiment 1 sample

[0054] Case-control study is the method used in this study. The research subjects of the experimental group were 4 patients with severe osteoarthritis who were hospitalized in the Department of Orthopedics of Peking Union Medical College Hospital in 2015 but had not yet received surgical treatment, and the research subjects of the control group were 4 relatively healthy people without osteoarthritis , the experimental group met the diagnostic criteria for osteoarthritis of the Orthopedic Branch of the Chinese Medical Association. The age of the experimental group was 58.5±2.0 years old (56-61 years old), and the age of the control group was 57.6±2.1 years old (55-61 years old). According to the "Adult Weight Judgment" standard formulated by the National Health and Family Planning Commission of the People's Republic of China in 2013, both groups selected overweight individuals with a BMI between 24 and 28. The experimental protoc...

Embodiment 2

[0067] Example 2 iTRAQ experiment screening differential proteins

[0068] 1. Instruments and reagents used

[0069] Vortex oscillator (Haimen Qilinbeier Instrument Manufacturing Co., Ltd., model: QL-901)

[0070] Centrifuge (Thermo, model: PICO17 )

[0071] Ultrasonic cell disruptor (Nanjing Xianou Instrument Manufacturing Co., Ltd., model: XO)

[0072] Microplate reader (Thermo, model: MμLtiskanMK3 )

[0073] Constant temperature incubator (Shanghai Pudong Rongfeng Scientific Instrument Co., Ltd., model: HH.S4)

[0074] Vacuum freeze dryer (Thermo, model: SPD2010-230 )

[0075] RIGOL L-3000 HPLC system (Beijing Puyuan Jingdian Technology Co., Ltd.), mobile phase A: 98% ddH2O, 2% acetonitrile (pH 10); mobile phase B: 98% acetonitrile, 2% ddH 2 O (pH 10) high performance liquid chromatography: (ThermoScientic EASY-nLC 1000System (Nano HPLC)), mobile phase A: 100% ultrapure water, 0.1% formic acid; mobile phase B: 100% acetonitrile, 0.1% formic acid mass spectrometry syste...

Embodiment 3

[0175] Example 3 western blot detection of GALNT18, IGFLR1 and MYDGF protein expression in patients with osteoarthritis

[0176] 1. Protein extraction

[0177] The protein extraction process was described in Example 2 simultaneously.

[0178] 2. Quantification of total protein using BCA protein concentration assay kit

[0179] Kangwei Century Trace BCA Protein Quantification Kit (Cat. No.: CW2011) was used, and the specific steps are shown in the instructions.

[0180] 3. SDS-polyacrylamide gel electrophoresis (SDS-PAGE)

[0181] 1. Protein sample denaturation:

[0182] a) According to the BCA protein concentration measurement results, the same mass of total protein extract was added to each gel sample well. Mix protein sample and protein loading buffer (5x) at a ratio of 0.25 μl protein loading buffer per 1 μl protein sample.

[0183] b) Heat at 100°C or in a boiling water bath for 3-5 minutes to fully denature the protein.

[0184] c) After cooling to room temperature,...

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Abstract

The invention discloses a marker for detecting osteoarthritis. The marker is GALNT18, IGFLR1 and MYDGF proteins. Further, the invention also provides application of the marker for detecting osteoarthritis to preparation of a reagent, a kit or a chip for diagnosing osteoarthritis. The three kinds of proteins with difference expression in the urine of a patient with the osteoarthritis are discovered; in addition, the application prospects in the early stage detection of the osteoarthritis are studied; the urine proteins relevant to the osteoarthritis provided by the invention can be used as early stage bio-marker for the osteoarthritis; a novel direction is provided for further studying the genetic molecular mechanism of osteoarthritis and exploring the medicine target points for osteoarthritis early stage prevention and treatment.

Description

technical field [0001] The invention relates to the technical field of biomedical detection, in particular to a marker for detecting osteoarthritis and its application. Background technique [0002] Osteoarthritis (OA) is a chronic degenerative joint disease that is caused by multiple factors and is very common in middle-aged and elderly people. The hip joint can also be found in the shoulder joint, wrist joint, ankle joint, etc. Osteoarthritis is characterized by wear and tear of articular cartilage, degeneration and bone hyperplasia around the joints, often accompanied by synovitis in the acute phase. According to the survey, the prevalence rate of osteoarthritis in the crowd over 60 years old is more than 50%, and the prevalence rate of the crowd over 75 years old is as high as 80%, and the disability rate of the disease can reach 53%. [0003] Osteoarthritis of the knee joint causes pain, swelling, stiffness, limited range of motion and dysfunction of the knee joint, w...

Claims

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Application Information

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IPC IPC(8): G01N33/564
CPCG01N33/564
Inventor 林进王炜范彧蔡思逸
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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