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DOCK10 gene for early diagnosis of myocardial infarction

A DOCK10, myocardial infarction technology, applied in disease diagnosis, microbial determination/examination, biochemical equipment and methods, etc., can solve the problem of unclear expression of related genes, achieve early diagnosis, reduce mortality, gene more effective diagnosis

Inactive Publication Date: 2018-11-13
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A variety of physiological processes accompany the occurrence of myocardial infarction, such as inflammation, abnormal calcium uptake, cell cycle abnormalities, peptide secretion, oxidative stress, apoptosis, etc., and the precise timing of these processes and the occurrence of myocardial ischemia are currently unknown. When some related genes are expressed is not clear and needs to be further explored

Method used

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  • DOCK10 gene for early diagnosis of myocardial infarction
  • DOCK10 gene for early diagnosis of myocardial infarction
  • DOCK10 gene for early diagnosis of myocardial infarction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Screening of genes differentially expressed in patients with myocardial infarction and normal people

[0035] 1. Research object

[0036] Select 6 hospitalized patients with myocardial infarction treated in the hospital as research objects, including 3 males and 3 females, with an average age of 56.00±8.75; 7 healthy people in the control group; each of the above-mentioned patients and healthy people who were invited to participate in the study All signed the informed consent.

[0037] 2. Sample collection and storage

[0038] On the day of admission, 8 mL of fresh sterile arterial blood was collected into EDTA anticoagulated purple tubes before coronary angiography. If it is not used immediately, the sample can be stored in a refrigerator at 4°C for 2 hours.

[0039] 3. Separation of PBMCs by Ficoll method

[0040] The following steps are all completed in the ultra-clean bench:

[0041] (1) Dilute the blood sample with equal volume of normal saline, add ...

Embodiment 2

[0063] Embodiment 2QPCR experiment verifies the genes differentially expressed in patients with myocardial infarction and normal people

[0064] 1. Research object:

[0065] The screening criteria were the same as in Example 1, 35 patients with myocardial infarction and 35 normal subjects.

[0066] 2. Extraction of total RNA in blood

[0067] Step is with embodiment 1.

[0068] 3. RT-PCR

[0069] (1)RT

[0070] RT reaction system (20μl):

[0071]

[0072]

[0073] RT reaction procedure:

[0074] 42°C 15min

[0075] 85℃ 5s

[0076] 4℃ ---

[0077] (2) qPCR

[0078] PCR reaction system (20μl):

[0079]

[0080] PCR reaction program:

[0081] Amplification procedure:

[0082] Stage 1 95°C 10min

[0083] Stage 2 95°C 10s

[0084] 55℃ 10s

[0085] Repeat 40 cycles

[0086] Three replicate wells were set for each sample, and the internal reference was GAPDH.

[0087] (3) Primers

[0088] The primer sequences of DOCK10 gene and GAPDH gene are as follows:

[...

Embodiment 3

[0102] Example 3 Western blot experiment to verify the expression products of differentially expressed genes in patients with myocardial infarction and normal people

[0103] 1. Research object: same as embodiment 2.

[0104] 2. Mononuclear cell isolation

[0105] Take 10ml of venous blood from patients with myocardial infarction and normal people, inject it into a sterile vial containing heparin, and shake it gently immediately after capping. Add an equal volume of HBSS (NaCl 8.0g, NaCl 2 HPO 4 0.132g, KH 2 PO 4 0.06g, KCl0.4g, phenol red 1ml, NaHCO 3 0.35g, D-glucose 1.0g, dissolved in 1000ml double distilled water), to reduce the aggregation of red blood cells. Draw 8ml of lymphocyte stratification solution into a 50ml centrifuge tube, slowly add the diluted blood along the tube wall, keep the interface clear, do not mix the two, centrifuge at 20°C 2000r / min for 30min, carefully absorb the stratification solution and plasma transfer The turbid off-white layer, tha...

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Abstract

The invention discloses a myocardial infarction diagnosis related gene. Genes in differential expression between a myocardial infarction patient and a normal person are screened through RNA-seq, and according to large sample QPCR verification, contents of a DOCK10 gene and DOCK10 protein in blood of the myocardial infarction patient are lower than those in blood of the normal person. Therefore, the DOCK10 gene and the DOCK10 protein can be used as biomarkers for diagnosis of myocardial infarction. By research results, a noninvasive method is provided for clinical early diagnosis of myocardialinfarction and suitable for clinical popularization.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis and relates to DOCK10 gene for early diagnosis of myocardial infarction. Background technique [0002] Myocardial infarction is a global public health problem that seriously threatens public health (J.Ross, Jr., A50-year research journey. From laboratory to clinic, Circulation journal: official journal of the Japanese Circulation Society, 73(2009 ) 3-12; D. Lloyd-Jones, R. Adams, M. Carnethon, G. De Simone, T. B. Ferguson, K. Flegal, E. Ford, K. Furie, A. Go, K. Greenlund, N. Haase , S. Hailpern, M. Ho, V. Howard, B. Kissela, S. Kittner, D. Lackland, L. Lisabeth, A. Marelli, M. McDermott, J. Meigs, D. Mozaffarian, G. Nichol, C . O'Donnell, V. Roger, W. Rosamond, R. Sacco, P Sorlie, R. Stafford, J. Steinberger, T. Thom, S. Wasserthiel-Smoller, N. Wong, J. Wylie-Rosett, Y. Hong, C. American Heart Association Statistics, S.). Several methodologies can be effectively applied to the treatment of m...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883C12Q2600/158G01N33/6893G01N2800/324
Inventor 李曙光靳传娣
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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