Application of MIF (migratory inhibitory factor) to preparation of product for diagnosing steroid-induced avascular necrosis of femoral head
A femoral head necrosis and hormone technology, applied in the field of biomedicine, can solve the theory of energy disorder, lipid metabolism, high pressure in the femoral head, vascular endothelial injury, bone marrow mesenchymal stem cell dysfunction, and can not be fully explained Pathological process of femoral head necrosis and other issues
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Embodiment 1
[0028] Example 1 The expression of the target gene MIF and the discovery of its regulator miR-146a
[0029] 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.
[0030] 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
[0033] Example 2 SANFH patient tissue and peripheral blood collection
[0034] 1. SANFH inclusion criteria
[0035] 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.
[0036] 2. SANFH exclusion criteria
[00...
Embodiment 3
[0042] Example 3 qPCR detection of MIF gene expression level
[0043] 1. RNA extraction
[0044] 1) RNA extraction sample processing
[0045] According to the standard in Example 2, 18 clinical samples were collected from the Department of Orthopedics, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, including 7 normal subjects and 11 SANFH patients. All have signed informed consent.
[0046] Mix EDTA anticoagulated whole blood with Trizol at a ratio of 1:1. If RNA extraction will not 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.
[0047] 2) RNA extraction method
[0048] 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).
[0049] 2.2) Preserve the sample in step 1), thaw at room temperature, and vigorously vortex to make it into a f...
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