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Heart failure detection device based on nucleic acid methylation change and application

A detection device, technology of nucleic acid methylation, applied in the field of medical detection, can solve the problem of not analyzing the change of methylation rate in heart failure

Inactive Publication Date: 2020-07-14
GUANGZHOU MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the existing technologies simply detect the methylation of genome-wide DNA in the blood (tissue) of patients with myocardial infarction or experimental animals, which has certain limitations, and does not analyze the change of methylation rate in the process of heart failure from the aspect of RNA. At the same time, there was no association analysis for heart failure disease tissue and peripheral blood

Method used

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  • Heart failure detection device based on nucleic acid methylation change and application
  • Heart failure detection device based on nucleic acid methylation change and application
  • Heart failure detection device based on nucleic acid methylation change and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] This example is used to illustrate the establishment of a myocardial infarction model.

[0076] The animals used in this experiment were SPF grade SD male rats. After the rats were anesthetized with 10% chloral hydrate (400mg / kg), they were fixed on the mouse board in the dorsal position, and the mouse board was tilted at 70°. Connect the rat tracheal tube to the small animal ventilator, the tidal volume is 2-4ml, and the respiratory rate is 60-70 times min -1 , The breathing ratio is 1:1. Carry out endotracheal intubation, observe the waveform displayed on the screen of the ventilator and the rising and falling state of the rat's chest. If the chest rises and falls regularly, it indicates that the insertion into the trachea has been successful. Open a longitudinal opening about 1.5 cm between the third and fourth ribs of the left chest, cut the third rib upwards, bluntly separate the muscle layers, open the chest cavity, use a chest expander to open the chest cavity,...

Embodiment 2

[0078] This example is used to illustrate the processing of samples.

[0079] (1) Myocardial tissue sample collection:

[0080] Rats in the sham group, 1 week, 4 weeks and 8 weeks after operation were euthanized to obtain samples. Rats were anesthetized with isoflurane, and the heart was taken out after opening the chest to collect blood. The blood in the heart was washed with sterile 4°C pre-cooled PBS buffer solution, blotted dry with filter paper, and the tissues in the infarcted area and non-infarcted area were separated. The rat hearts in the experimental group were collected. Myocardial tissue in the infarct border area, and the control group was obtained from the same area of ​​the heart. After rapid freezing in liquid nitrogen, they were stored at -80°C.

[0081] (2) Extraction of RNA and DNA from myocardial tissue and blood samples:

[0082] Rat heart blood was drawn into sodium heparin anticoagulant tubes, and lymphocytes were separated using rat peripheral blood ...

Embodiment 3

[0094] This example is used to illustrate the instrument conditions.

[0095] (1) Tandem mass spectrometry conditions:

[0096] Agilent Jet Stream Ion Focusing Electrospray Ion Source (AJS ESI), positive ion mode, multiple reaction monitoring mode (Multiple Reaction Monitoring, MRM) quantitative; capillary voltage (Capillary) 3000V, positive ion mode, nozzle voltage 500V, drying gas temperature 300°C, drying gas flow rate 8.0L min -1 , atomizing gas pressure 35psi, sheath gas temperature 350°C, sheath gas flow rate 11.0L min -1 .

[0097] (2) Ultra-high performance liquid chromatography conditions:

[0098] EclipsePlusC18 RRHD (1.8μm, 2.1mm×50mm); mobile phase: A ultrapure water containing 0.1% formic acid, mobile phase: B is methanol containing 0.1% formic acid; flow rate is 0.2mL min -1 , column temperature 35°C, injection volume 5μL; Gradient elution: 0-1.35min, 0.05B; 1.35-2.60min, 0.05-0.95B; 2.6-3.5min, 0.95B; 3.5-3.51min, 0.95-0.05B ; 3.51-5.5min, 0.05B, each compo...

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Abstract

The invention provides a heart failure detection device based on nucleic acid methylation change and application. The device comprises an RNA and DNA extraction mechanism, an RNA and DNA enzymolysis mechanism and a methylated RNA and DNA determination mechanism. The invention finds that the methylation rates in myocardial tissues and peripheral blood of a myocardial infarction mouse both have an increasing trend, and the trends are consistent, so that peripheral blood lymphocytes are prompted to possibly become good substitutes for the myocardial tissues, and the methylation rates are positively correlated with the severity of myocardial infarction. The invention preliminarily shows that DNA and RNA methylation may have a certain potential effect in the occurrence and development of heartfailure, and may be used as a reference index for clinical heart failure diagnosis.

Description

technical field [0001] The invention belongs to the field of medical detection, and in particular relates to a heart failure detection device and application based on nucleic acid methylation changes. Background technique [0002] In recent years, most studies on heart failure-specific methylation changes have focused on gene and promoter-specific changes. Measuring the potential role in heart disease prevention, as well as monitoring the therapeutic effects of therapeutics being evaluated in human clinical trials, remains a valuable technical tool and a good biomarker for monitoring large-scale epigenetic events in organisms thing. [0003] Emerging evidence suggests that post-transcriptional modification of mRNA is critical for its stability, nuclear export, cell division, translation into protein, degradation, and stem cell pluripotency. In eukaryotic mRNA, two RNA modifications have been extensively studied: N6-methyladenosine (m6A) and 5-methylcytosine (m5C). Studies...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/40C12M1/00
CPCC12Q1/68C12Q1/6806C12Q2521/327C12Q2565/125C12Q2565/627C12Q2565/137
Inventor 余细勇林忠晓常继硕张灵敏
Owner GUANGZHOU MEDICAL UNIV
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