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Application of miR-146a in preparation of products for diagnosing steroid-induced femoral head necrosis

A 1.mir-146a, femoral head necrosis technology, applied in the field of biomedicine, can solve the energy disorder theory, lipid metabolism theory, high pressure theory in the femoral head, vascular endothelial injury theory, bone marrow mesenchymal stem cell dysfunction theory , can not fully explain the pathological process of femoral head necrosis and other problems, to achieve the effect of rapid and effective early detection

Inactive Publication Date: 2019-05-17
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on SANFH has always been a hot spot in orthopedic research. Because of its complex etiology, there are many hypotheses about the pathogenesis of SANFH, such as coagulation disorder theory, lipid metabolism theory, femoral head high pressure theory, vascular endothelial injury theory, bone marrow Mesenchymal stem cell dysfunction theory, etc., although these studies have some clinical and experimental research basis, they cannot fully explain the entire pathological process of femoral head necrosis

Method used

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  • Application of miR-146a in preparation of products for diagnosing steroid-induced femoral head necrosis
  • Application of miR-146a in preparation of products for diagnosing steroid-induced femoral head necrosis
  • Application of miR-146a in preparation of products for diagnosing steroid-induced femoral head necrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 The expression of the target gene MIF and the discovery of its regulator miR-146a

[0032] The inventors detected the macrophage migration inhibitory factor (macrop hagemigration inhibitory factor, MIF) in the peripheral blood of multiple SANFH patients, and found that the expression levels of MIF gene and protein in the peripheral blood of SANFH patients were significantly lower than normal.

[0033] In order to further explore the low expression of MIF in the peripheral blood of SANFH patients and the mechanism affecting the occurrence and development of SANFH, the inventors used software to search for other proteins or RNAs that could interact with MIF. miRNA mainly binds to the 3'UTR region of the target gene through its own seed sequence, degrades the target gene RNA or inhibits its translation. Studies have shown that miRNA affects the occurrence and development of diseases by regulating the expression of target genes, and participates in the signaling n...

Embodiment 2

[0036] Example 2 SANFH patient tissue and peripheral blood collection

[0037] 1. SANFH inclusion criteria

[0038] 1) Sign the informed consent form; 2) Have complete clinical data and imaging data before surgery; 3) High-dose hormone pulse therapy > 3 days or conventional dose hormone therapy > 3 weeks; 4) Pain in the groin radiating to the thigh, Slow progressive aggravation, aggravated pain at night; 5) Pain not relieved by drug treatment; 6) Imaging imaging of patients with femoral head necrosis of stage II and above showed sclerosis, cystic lesions, femoral head collapse, femoral cartilage fractures and spots Sclerotic lesions; 7) T1-weighted imaging showed femoral head residual bone plaque line, adjacent bone plaque line showed low signal area, and T1-weighted imaging showed double-line sign. 8) At the same time, non-necrosis of the femoral head patients who received hormone therapy were selected as the normal control group.

[0039] 2. SANFH exclusion criteria

[00...

Embodiment 3

[0045] Example 3 qPCR detection of expression levels of MIF gene and miR-146a

[0046] 1. RNA extraction

[0047] 1) RNA extraction sample processing

[0048] According to the standard of Example 2, 32 clinical samples were collected from the Department of Orthopedics, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, including 6 normal subjects and 26 patients. All have signed informed consent.

[0049] Mix EDTA anticoagulated whole blood with Trizol at a ratio of 1:1. If RNA extraction is not to be performed within a short period of time, the sample can be quickly frozen in liquid nitrogen for 30s-1min, and stored at -80°C for short-term storage.

[0050] 2) RNA extraction method

[0051] 2.1) Chloroform, isopropanol, ethanol (all of which are pure for chemical analysis), and small desktop centrifuges need to be pre-cooled in advance (10-15min).

[0052] 2.2) Preserve the sample in step 1), thaw at room temperature, and vortex vigorously to make...

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Abstract

The invention discloses application of miR-146a as a marker in preparation of products for diagnosing steroid-induced femoral head necrosis. According to the application, expression of the miR-146a inbiological samples with steroid-induced femoral head necrosis is raised. According to the application, the products comprises a kit for diagnosing steroid-induced femoral head necrosis, and the kit comprises preparations for detecting the expression level of the miR-146a. By the adoption of the miR-146a to detect steroid-induced femoral head necrosis, not only can early detection be rapidly and effectively achieved, but also therapeutic targets and important evidence are provided for clinical application such as gene therapy and drug therapy.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of miR-146a in the preparation of products for diagnosing steroid-induced necrosis of the femoral head. Background technique [0002] Osteonecrosis of femoral head (ONFH), also known as avascular necrosis (AVN), is a common and refractory orthopedic disease that occurs in young and middle-aged people, with a high morbidity rate. National epidemiological studies in Korea and Japan suggest that the annual incidence of femoral head necrosis is as high as 28.9 / 100,000. In my country, the number of ONFH patients who need treatment is 5-7.5 million each year, and the number of new patients is 150,000-200,000 each year. Most of the patients are young adults aged 37-45. The number of males is more than that of females. The ratio of male to female in my country It is 3.8:1. Steroid-induced avascular necrosis of the femoral head (SANFH) accounted for 66.7% of non-tra...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
Inventor 翁习生边焱焱项永波李曾程开源
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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