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Application of Molecular Markers in Male Osteoporosis

A technology for osteoporosis and men, applied in the field of biomedicine, can solve the problem of unclear pathogenesis of osteoporosis and achieve the effect of improving the quality of life

Inactive Publication Date: 2020-09-29
王丽 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although current studies suggest that the onset of osteoporosis is related to age, gender, and is closely related to genetic factors and environmental factors, the pathogenesis of osteoporosis is still unclear. Early diagnosis, treatment monitoring and prognosis evaluation of osteoporosis Plays an important role in the prevention and treatment of osteoporosis

Method used

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  • Application of Molecular Markers in Male Osteoporosis
  • Application of Molecular Markers in Male Osteoporosis
  • Application of Molecular Markers in Male Osteoporosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1 Screening for Gene Markers Related to Osteoporosis

[0078] 1. Sample collection

[0079] The blood of 6 male patients with osteoporosis and 6 healthy people were collected respectively. None of the above samples had a history of smoking, drinking, maternal family history, history of coffee or carbonated drinks, lack of physical activity, diseases affecting bone metabolism, or use of related drugs . The above-mentioned samples were obtained with the informed consent of the patients, and all the above-mentioned samples were obtained with the consent of the organizational ethics committee.

[0080] 2. RNA sample preparation and quality analysis

[0081] 2.1 Preparation of RNA samples

[0082] Total RNA was extracted using the RNA extraction kit from Promega. Specific steps are as follows:

[0083] 1) Take 1ml of whole blood collected in a test tube treated with heparin or EDTA, and put it into a sterile centrifuge tube;

[0084] 2) Centrifuge at 3000rpm (4...

Embodiment 2

[0108]Example 2 QPCR verification of differential expression of BTNL3 gene

[0109] 1. Large-sample QPCR verification of differential expression of BTNL3 gene. According to the sample collection method in Example 1, 80 cases of blood from osteoporosis patients and 80 healthy people were selected.

[0110] 2. RNA extraction The steps are as described in Example 1.

[0111] 3. Reverse transcription

[0112] Use the FastQuant cDNA First Strand Synthesis Kit (Cat. No.: KR106) for reverse transcription of mRNA

[0113] 4. QPCR

[0114] SuperReal PreMix Plus (SYBR Green) (product number: FP205) was used for amplification, and the experimental operation was performed according to the product manual.

[0115] 1) Primer design

[0116] QPCR amplification primers were designed according to the coding sequences of BTNL3 gene and GAPDH gene in Genebank, and synthesized by Biomed Biotech. The specific primer sequences are as follows:

[0117] BTNL3 gene:

[0118] The forward primer...

Embodiment 3

[0129] Example 3 Verification of the differential expression of BTNL3 at the protein level

[0130] 1. Extract each histone according to the instructions of the RIPA protein lysate kit, and use the BCA protein concentration assay kit to detect the protein concentration in the sample.

[0131] 2. The change of BTNL3 protein was detected by conventional Western-blot method. The experiments in each group were repeated 3 times, and the absorbance of BTNL3 protein band was quantitatively analyzed with β-actin as the internal reference. The expression amount was represented by the ratio of BTNL3 protein / β-actin absorbance .

[0132] 3. Results

[0133] The result is as figure 2 As shown, compared with healthy people, the protein level of BTNL3 in male osteoporosis patients was significantly up-regulated, and the difference was statistically significant (P<0.05).

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Abstract

The invention discloses an application of a molecular marker in male osteoporosis. The molecular marker is BTNL3. An experiment proves that the expressions of BTNL3 and encoded protein thereof in an osteoporosis patient are up-regulated so that the osteoporosis can be early diagnosed in the manner of detecting the expression level of BTNL3.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of molecular markers in male osteoporosis, specifically the molecular marker is BTNL3. Background technique [0002] Osteoporosis (osteoporosis) is a systemic bone disease in which bone density and bone quality decrease due to various reasons, bone microstructure is destroyed, bone fragility increases, and fractures are prone to occur. Osteoporosis is divided into primary and secondary two categories. The occurrence and development of primary osteoporosis are closely related to endocrine disorders, systemic nutritional status, immune function, and genetics. [0003] With the rapid economic development and the acceleration of industrialization, the threat of non-communicable diseases to human health is increasing. Osteoporosis and osteoporotic fractures are ruthless killers that endanger health, and have been widely regarded as the One of the major health problems of huma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883G01N33/68
Inventor 王丽马青王玉功
Owner 王丽
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