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Application of miRNA marker hsa-miR-486-5p

A marker, coal worker's pneumoconiosis technology, applied in the fields of genetic engineering and life sciences, to improve sensitivity and specificity, quickly and accurately grasp the patient's condition, and promote the effect of diagnosis

Inactive Publication Date: 2014-11-26
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are currently no relatively stable biomarkers reported for auxiliary early diagnosis of coal worker pneumoconiosis. If serum miRNAs specifically or abnormally expressed in coal worker pneumoconiosis can be screened as biomarkers, and corresponding auxiliary early diagnosis kits can be developed , will be a powerful impetus to the current situation of pneumoconiosis diagnosis in my country

Method used

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  • Application of miRNA marker hsa-miR-486-5p
  • Application of miRNA marker hsa-miR-486-5p
  • Application of miRNA marker hsa-miR-486-5p

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Effect test

Embodiment 1

[0047] Establishment of mouse lung fibrosis animal model induced by silica dust: male C57BL / 6 mice weighing 20-25 g at 6-8 weeks were selected, and the mice were treated with tracheal perfusion of silica suspension. Mice were treated on days 7, 14, 28, and 56, and lung tissue, serum, and bronchial lavage fluid were preserved. The success of the silica dust-induced fibrosis model was observed by pathological sections of mouse lung tissue.

[0048]

Embodiment 2

[0050] According to the pathological sections of lung tissue, one mouse lung tissue was selected from the control group, and the silica dust treatment group on the 3rd, 7th, 14th, 28th, and 56th day to extract RNA, and the Affymetrix miRNA 3.0 chip was used for detection, and the miRNA chip results were analyzed. Comparing the silica dust treatment groups with the control group at different periods, a total of 259 miRNAs were screened out that were up-regulated or down-regulated by at least two times compared with the control group in at least one treatment group. Among them, there were 108, 74, 37, 89 and 130 miRNAs in the 3-day, 7-day, 14-day, 28-day, and 56-day groups after silica dust treatment compared with the control group, respectively. Further analysis found that compared with the control group, there were 62 miRNAs that were differentially expressed twice or more in at least three groups, of which 17 had an original expression value greater than 100, including 9 miRNA...

Embodiment 3

[0053] qRT-PCR experiments of miRNAs in animal models of pulmonary fibrosis: According to the results of the above-mentioned Affymetrix miRNA 3.0 chip, mmu-miR-1a, mmu-miR-133a, mmu-miR-133b, mmu-miR-146b, mmu-miR- 223, mmu-miR-449a, mmu-miR-2137, mmu-miR-5128, mmu-miR-21, mmu-miR-122, mmu-miR-351-star, mmu-miR-193-star, mmu- A total of 17 miRNAs including miR-574-3p, mmu-miR-3107, mmu-miR-151-3p, mmu-miR-455, and mmu-miR-486-5p were detected by qRT-PCR, with 5 mice in each experimental group . Strict quality control was implemented throughout the research process, each sample was tested three times in a row, and all samples were blinded, that is, completed without knowing the background of the sample to avoid bias.

[0054] (1) Preparation of RNA samples: ① Place the sterilized tissue homogenizer on ice, put 100mg of mouse lung tissue into it, and then add 1mL Trizol to grind until homogenized; ② Transfer the homogenate to a 1.5mL EP tube , add 200 μL of chloroform (trichlo...

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Abstract

The invention provides an application of miRNA marker hsa-miR-486-5p. A serum micro ribonucleic acid marker provided by the invention as a CWP early diagnosis marker has the advantages that serum miRNAs are novel biomarkers, are different from traditional biomarkers, and have the advantages of being stable, minimally invasive, easy to detect, accurate to quantify and the like, the sensitivity and the specificity of disease diagnosis are greatly improved, a new situation is created for early diagnosis of CWP by successful development of micromolecular RNA biomarkers, and reference is provided for development of other disease biomarkers.

Description

[0001] technical field [0002] The invention belongs to the field of genetic engineering and life sciences, and discloses the application of a miRNA marker hsa-miR-486-5p. [0003] Background technique [0004] Pneumoconiosis is a systemic disease mainly caused by diffuse fibrosis of lung tissue caused by long-term inhalation of productive dust in occupational activities. At present, pneumoconiosis is still the most extensive and serious occupational disease in my country. Data from the National Occupational Disease Reporting System shows that in 2012, a total of 24,206 new cases of pneumoconiosis were reported in the country, of which 12,405 were coal worker pneumoconiosis and silicosis. and 10592 cases, accounting for 51.25% and 43.76% of pneumoconiosis cases respectively. The reported cases of pneumoconiosis accounted for 88.28% of the total reported cases of occupational diseases in 2012. Once pneumoconiosis occurs, it is difficult to cure and seriously damages health...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 倪春辉吉晓明吴白群韩如慧杨京津王婷
Owner NANJING MEDICAL UNIV
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