Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of DACT2 gene in preparation of product for diagnosis and treatment of atrial fibrillation

A product and gene technology, applied in the field of application of DACT2 gene in the preparation of atrial fibrillation diagnosis and treatment products, can solve the problem of lack of diagnostic methods

Active Publication Date: 2019-10-08
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the clinical diagnosis of atrial fibrillation mainly relies on the diagnosis of electrocardiogram, but when the above methods are used to diagnose, most of the patients have long-term persistent atrial fibrillation, and there is still a lack of a specific and sensitive diagnosis for early paroxysmal atrial fibrillation method

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of DACT2 gene in preparation of product for diagnosis and treatment of atrial fibrillation
  • Application of DACT2 gene in preparation of product for diagnosis and treatment of atrial fibrillation
  • Application of DACT2 gene in preparation of product for diagnosis and treatment of atrial fibrillation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] 1. Sample collection

[0057] A total of 28 patients who underwent valve replacement surgery due to valvular heart disease were selected, including 18 patients with atrial fibrillation (AF) and 10 patients with sinus rhythm (SR). Right atrial appendage tissue samples were obtained from all patients during the operation. The acquisition of all the above specimens was approved by the ethics committee of the organization.

[0058] 2. The specific steps of Masson staining and immunohistochemical staining are as follows:

[0059] (1) Fix fresh tissue specimens in 4% paraformaldehyde, carry out paraffin-embedded sections, and save them for future use.

[0060] (2) Masson staining

[0061] A. Paraffin sections are dehydrated: Put the specimen sections in turn in xylene Ⅰ 20min-xylene Ⅱ 20min-absolute ethanol Ⅰ 10min-absolute ethanol Ⅱ 10min-95% alcohol 5min-90% alcohol 5min-80% alcohol 5min-70% alcohol 5min-wash with distilled water.

[0062] B. Hematoxylin staining of cel...

Embodiment 2

[0090] Example 2 DACT2 gene overexpression

[0091] 1. Obtain primary atrial fibroblasts from adult SD rats, and incubate at 37°C, 5 %CO 2 cultured in an incubator. The medium was changed once every 2-3 days, and 0.25% trypsin was used for routine digestion and passage.

[0092] 2. Overexpression of DACT2 gene

[0093] (1) Construction of DACT2 gene overexpression vector

[0094] Ad5-DACT2-eGFP virus and Ad5-CON177-eGFP empty virus overexpressing DACT2 gene were constructed by Shanghai Jikai Gene Chemical Technology Co., Ltd. using Ad-CMV-eGFP vector.

[0095] (2) Transfection

[0096] The cells were planted in a 6-well plate, and after 24 hours of culture, the fresh medium containing the virus multiplicity of infection (MOI) required for each cell (Ad5-DACT2-eGFP virus or Ad5-CON177-eGFP empty virus) was replaced for infection. After 18 hours, aspirated Remove the medium containing the virus and replace with fresh medium.

[0097] (3) Immunofluorescence detection

[0...

Embodiment 3

[0105] Example 3 Effect of DACT2 Gene Overexpression on SD Rat Atrium Primary Fibroblasts

[0106] 1. RT-PCR method was used to detect the effect of DACT2 gene on the expression levels of collagen I and II in SD rat atrial primary fibroblasts.

[0107] 2. The specific steps of RT-PCR are as follows:

[0108] After Ad-DACT2-eGFP infected H9C2 cells for 48 hours, RNA was extracted by Trizol method, and cDNA was synthesized by PrimeScript TM RT Master Mix (Perfect Real Time) kit, conditions: 37°C, 15min; 85°C, 5s; store at 4°C. Real-time PCR uses TB Green TM Premix Ex Taq II (TliRNAseH Plus) kit, the detection instrument is LightCycler480 fluorescent quantitative PCR instrument, the reaction conditions are: pre-denaturation: 95°C, 30s; PCR reaction: 95°C, 5s; 60°C, 20s; 40 cycles; melting curve Settings: 95°C, 15s; 60°C, 1min; 95°C, 15s.

[0109] β-actin was selected as the internal reference gene, ultrapure water was used to replace the template DNA for the blank control, ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an application of DACT2 gene in preparation of a product for diagnosis and treatment of atrial fibrillation. The application finds for the first time that DACT2 gene expressionis associated with atrial fibrillation, by detecting the expression of DACT2 in the myocardial tissue of a subject, the subject can be judged whether the subject has atrial fibrillation or not, or whether the subject is at risk of having atrial fibrillation or not, so that a clinician can be instructed to provide the subject with a prophylactic scheme or a treatment scheme. The invention discovers a new molecular marker of atrial fibrillation-DACT2 gene, which is more timely, more specific and more sensitive than traditional detection methods, early diagnosis of atrial fibrillation can be achieved, and the mortality of atrial fibrillation is reduced.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of DACT2 gene in the preparation of atrial fibrillation diagnosis and treatment products. Background technique: [0002] Atrial fibrillation (atrial fibrillation, AF, atrial fibrillation) is one of the most common arrhythmias, often leading to heart failure, stroke and other embolism symptoms, increasing the mortality rate of hospitalized patients and the incidence of complications. Symptoms of AF can be asymptomatic, palpitations, or serious complications, including frequent hospitalization, hemodynamic instability, and thromboembolism. Atrial fibrillation significantly increases the risk of stroke in the population, which is about 5 times that of normal people, and is positively correlated with the increase of age. [0003] Clinically, atrial fibrillation is generally divided into paroxysmal atrial fibrillation, persistent atrial fibrillation, and long...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11G01N33/68A61K45/00A61P9/06
CPCC12Q1/6883G01N33/6893A61K45/00A61P9/06G01N2800/326C12Q2600/158
Inventor 吴钟凯黄少杰侯健龙雁姚尖平陈光献梁孟亚岳媛
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com