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miRNAs marker used for diagnosing carotid atherosclerotic plaques and applications thereof

A technology of atherosclerosis and markers, applied in the field of laboratory medicine

Pending Publication Date: 2019-09-13
JINHUA MUNICIPAL CENT HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although miRNAs have been initially studied as biomarkers of atherosclerosis, systematic studies of miRNA biomarkers related to different types of plaques (stable plaques, vulnerable plaques, and responsible plaques) have not been reported.

Method used

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  • miRNAs marker used for diagnosing carotid atherosclerotic plaques and applications thereof
  • miRNAs marker used for diagnosing carotid atherosclerotic plaques and applications thereof
  • miRNAs marker used for diagnosing carotid atherosclerotic plaques and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Collection and arrangement of serum samples from patients with carotid atherosclerosis and corresponding healthy controls

[0055] A total of 35 patients in the cerebral infarction group were selected from the Neurology Department of our hospital from September 2016 to March 2017, including 24 males and 11 females, aged 50-84 (69.3±10.6) years; There were 41 patients with unstable carotid artery plaques confirmed by Le color Doppler ultrasound (unstable plaque group), 24 males and 17 females, aged 45-90 (66.0±11.2) years; patients with stable plaques (stable plaque group) 28 cases, 16 males and 12 females, aged 58-82 (68.3±5.8) years old; another 30 cases of healthy people were selected as the healthy control group, 16 males, 14 females, aged 54-78 (66.7±7.2 )age. The diagnosis of patients in the cerebral infarction group was in line with the 2014 Chinese guidelines for the diagnosis and treatment of acute ischemic stroke, and the cerebral infarction of the i...

Embodiment 2

[0057] Example 2 Extraction of Carotid Atherosclerosis and Corresponding Healthy Serum Total RNA

[0058] (1) The serum sample obtained in Example 1 was taken out at -80°C, slowly dissolved on ice, 200 μl of serum was transferred to a centrifuge tube, 600 μl of Trizol was added, and placed in a vortex shaker for 20 seconds, then static at room temperature Set aside for 5 minutes to fully lyse the nucleoprotein complex to obtain a lysate;

[0059] (2) Add 200 μl of chloroform to the lysate prepared in step (1), shake vigorously for 15 seconds, and let stand at room temperature for 3 minutes to obtain mixture A;

[0060] (3) Turn on the centrifuge in advance to pre-cool it to 4°C, and centrifuge the mixture A prepared in step (2) at 12000g for 15min;

[0061] (4) Use a pipette to transfer the supernatant liquid after centrifugation in step (3) to a new centrifuge tube, add 0.6 to 1 times the volume of isopropanol and mix well, and put the mixed sample at 15-20°C Incubate for 1...

Embodiment 3

[0067] Total RNA quantification and quality detection of embodiment 3

[0068] NanDrop2000 was used to detect the content and quality of RNA in serum samples to ensure the reliability of experimental data. The specific detection method for total RNA quantification is as follows: Measure the OD value of RNA at 260nm and 280nm wavelengths on a NanDrop2000 spectrophotometer, and obtain the ratio of A260 / A280, when the ratio is between 1.8 and 2.0, it indicates that the purity of the total RNA is relatively high. Well, if it is less than 2.0, it means that there are residual salt ions and contamination of small molecular magazines, and when it is greater than 2.0, it means that there may be degradation of total RNA.

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Abstract

The invention discloses an miRNAs marker used for diagnosing carotid atherosclerotic plaques and applications thereof. The expression levels of miR-21, miR-155, miR-221 and miR-222 in an unstable plaque with cerebral infarction group, an unstable plaque group and a stable plaque group can be obviously enhanced compared with a healthy control group; the expression level of miR-126 in the unstable plaque with cerebral infarction group and the unstable plaque group can be obviously reduced compared with the stable plaque group; and the expression level of the miR-126 in the unstable plaque with cerebral infarction group can also be obviously reduced compared with the unstable plaque group. The above five miRNAs are closely related to the growth, instability and rupture evolution processes ofthe plaques, so that the marker can be applied to the assessment of the stability of carotid atherosclerotic plaques and the screening of cerebral apoplexy high-risk population.

Description

technical field [0001] The invention relates to the technical field of laboratory medicine, in particular to a group of diagnostic marker miRNAs for carotid atherosclerotic plaques and applications thereof. Background technique [0002] According to the third national survey of causes of death by the Ministry of Health, cerebrovascular disease has become the first cause of death in Chinese people, of which cerebral infarction accounts for about 75%, which has high incidence, high recurrence rate, high disability rate, high The characteristics of the "five highs" of fatality rate and high cost have caused serious burdens to individuals, families and society. Our country spends as much as 40 billion yuan on stroke medical expenses every year. Embolus caused by carotid atherosclerotic plaque rupture is the main cause of cerebral infarction, and its formation is not only related to the size of the plaque, but also significantly related to the stability of the plaque. In recent...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/178
Inventor 陈红芳程兴潘小玲
Owner JINHUA MUNICIPAL CENT HOSPITAL
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