Application of smchd1 in preparation of products for diagnosing coronary heart disease
A technology for coronary heart disease and products, which is applied in material inspection products, biochemical equipment and methods, and microbial determination/inspection, etc., can solve the problems of lack of disease registration and classification system, inability to obtain very accurate epidemiological data, etc.
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Embodiment 1
[0051] Example 1 Differential expression of SMCHD1 gene
[0052] 1. Research object:
[0053] Collect 5 cases of patients with coronary heart disease, 5 cases of normal people.
[0054] Inclusion criteria for the coronary heart disease group: according to the diagnostic criteria for ischemic heart disease formulated by the World Health Organization, patients with coronary heart disease who were confirmed by coronary angiography to have stenosis of one or more vessels ≥ 50% were selected. The inclusion criteria for the control group are: 1) The age, gender, and ethnicity are all matched with the CAD group during the epidemiological investigation, and there is no coronary heart disease after questionnaire survey, physical examination, cardiac ultrasound examination, electrocardiogram examination and related laboratory tests. Physical examination of clinical manifestations and major risk factors; 2) Inpatient physical examination and coronary angiography to exclude coronary hear...
Embodiment 2
[0082] Example 2 QPCR experiment verification of genes differentially expressed in patients with coronary heart disease and normal people
[0083] 1. Research object:
[0084] The screening criteria were the same as in Example 1, 50 patients with coronary heart disease and 50 normal subjects.
[0085] 2. Extraction of total RNA in blood
[0086] Step is with embodiment 1.
[0087] 3. Reverse transcription
[0088] 1 μg of total RNA was reverse-transcribed to synthesize cDNA using reverse transcription buffer. Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, add the following components to the PCR tube respectively: DEPC water, 5× reverse transcription buffer, 10mmol / L dNTP, 0.1mmol / l DTT, 30μmmol / l Oligo dT, 200 U / μl M-MLV, template RNA. Incubate at 42°C for 1 hour, then centrifuge briefly at 72°C for 10 minutes.
[0089] 4. QPCR
[0090] (1) Primer design
[0091] QPCR amplification primers were designed according to the coding sequences o...
Embodiment 3
[0108] Example 3 Detection of differential expression of SMCHD1 protein
[0109] 1. Research object
[0110] With embodiment 2.
[0111] 2. Mononuclear cell isolation
[0112] Aseptically collect 10ml of venous blood, 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, KCl 0.4g, 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 The turbid off-white layer at the transition site, that is, the lymphocyte layer, was added to another centrifuge tube, washed twice with 5 times the volume of HBSS, and c...
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