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IncRNA markers of cerebral arterial thrombosis

An ischemic stroke and marker technology, applied in the field of biomedicine, can solve problems such as functions and mechanisms of action that need to be further explored, and achieve the effects of high sensitivity, improved accuracy, and improved quality of life

Active Publication Date: 2017-04-05
THE FIRST AFFILIATED HOSPITAL OF SHANTOU UNIV MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although lncRNA-related research has made progress in recent years, its function and mechanism of action still need to be further explored.

Method used

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  • IncRNA markers of cerebral arterial thrombosis
  • IncRNA markers of cerebral arterial thrombosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Screening for Gene Markers Related to Ischemic Stroke

[0041] 1. Sample collection

[0042] The blood of 10 normal people and the blood of patients with ischemic stroke were collected respectively, and the patients gave informed consent, and all the above-mentioned samples were obtained with the consent of the organizational ethics committee.

[0043] 2. Preparation of RNA samples

[0044] 1) After clinical serum samples are collected, centrifuge at 12,000 rpm for 10 minutes at high speed, repeat twice, and store the obtained serum samples at -80°C;

[0045] 2) Thaw frozen serum samples at 4°C;

[0046] 3) Pipette 250 μl serum sample to 1.5 ml EP tube, add 750 μl Trizol LS Reagent, mix by pipetting, and let stand for 5 minutes;

[0047] 4) Add chloroform (CHCl 3 : trizol250μ1:750μ1), mix upside down, let stand for 15min;

[0048] 5) 4°C, 12000rpm, centrifuge for 15min;

[0049] 6) Carefully draw the upper liquid into a new EP tube;

[0050] 7) Add an a...

Embodiment 2

[0065] Example 2 QPCR sequencing to verify the differential expression of LINC00189 gene

[0066] 1. Large-sample QPCR verification of differentially expressed lncRNAs. According to the sample collection method in Example 1, 100 blood samples from normal subjects and 100 patients with ischemic stroke were selected.

[0067] 2. The RNA extraction steps are the same as in Example 1.

[0068] 3. Reverse transcription:

[0069] (1) Reverse transcription reaction:

[0070] Using a 25 μl reaction system, take 1 μg total RNA for each sample as template RNA, and add the following components to a PCR tube: DEPC water, 5× reverse transcription buffer, 10 mM dNTP, 0.1 mM DTT, 30 μM OligodT, 200 U / μl M- MLV, template RNA. Incubate at 42°C for 1h, then centrifuge briefly at 72°C for 10min. (2) Primer design:

[0071] The primer sequence of LINC00189 gene is:

[0072] Forward primer: 5'-TCTTTGATTGACTGATTCTTT-3'(SEQ ID NO.3)

[0073] Reverse primer: 5'-TTGAGGAACACATCTGAA-3' (SEQ ID N...

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Abstract

The invention discloses IncRNA markers of cerebral arterial thrombosis. The IncRNA markers are LINC00189 and LOC105376014, wherein the expression of the LINC00189 is up-regulated in a patient with cerebral arterial thrombosis, and the expression of the LOC105376014 is down-regulated in the patient with cerebral arterial thrombosis. The IncRNA markers can be used for early diagnosis of cerebral arterial thrombosis and has relatively high sensitivity and specificity.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to lncRNA markers of ischemic stroke. Background technique [0002] As a major global public health problem, stroke is a sudden-onset cerebral blood circulation disorder and one of the major diseases that endanger human health. Stroke has become the second leading cause of death and the leading cause of disability worldwide. More than two-thirds of stroke-related deaths occur in developing countries. Statistics show that about 230 million people in my country suffer from cardiovascular disease, and more than 7 million stroke patients. A prospective cohort study involving 169,871 adults aged 40 and over showed that stroke ranks as the third cause of death in men and the second cause of death in women, and the mortality rate of stroke in men is 310.5 / 100,000 person-years , accounting for 21.6% of the total deaths, female stroke mortality rate was 242.3 / 100,000 person-years, accounting for 2...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/113
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 何文贞魏敦灿汤丹灵蔡德
Owner THE FIRST AFFILIATED HOSPITAL OF SHANTOU UNIV MEDICAL COLLEGE
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