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Diagnostic marker for abdominal aortic aneurysm

A technology for abdominal aortic aneurysm and drug, which is applied in the field of tumor diagnosis with the detection of GBP5 abnormality as a means, can solve the problem that the pathogenesis of abdominal aortic aneurysm is not completely clear.

Inactive Publication Date: 2017-08-04
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the specific pathogenesis of abdominal aortic aneurysm is not completely clear. Existing reports have shown that its pathogenesis is closely related to factors such as genetic factors, inflammation, protease degradation, and smooth muscle cell apoptosis. different mutations

Method used

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  • Diagnostic marker for abdominal aortic aneurysm
  • Diagnostic marker for abdominal aortic aneurysm
  • Diagnostic marker for abdominal aortic aneurysm

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Gene Chip Screening for Differentially Expressed Genes

[0060] 1. Tissue acquisition and processing: Collect 2 live specimens of the middle layer of the AAA artery wall and 2 copies of the normal infrarenal aortic control tissue. Under sterile and RNase-free conditions, cut the aortic wall at about 4 cm below the flat renal artery. Rinse with sterilized normal saline to remove blood stains, remove arterial intima and intima from the collected tissues, and quickly (<5 min) freeze them in liquid nitrogen for later use.

[0061] 2. RNA extraction and cDNA synthesis

[0062] The aneurysm tissues preserved in liquid nitrogen were placed in a ceramic mortar under the low temperature environment of liquid nitrogen and crushed into powder, added Trizol reagent to homogenate, centrifuged, and the supernatant was extracted twice with 1:1 acidic phenol-chloroform After extraction with sodium acetate and 5:1 acidic phenol-chloroform, an equal volume of isopropanol preci...

Embodiment 2

[0071] Example 2 Verification of Differentially Expressed Genes in Large Samples

[0072] The expression in abdominal aortic aneurysm tissue was selected to select the gene chip prompting differential expression of GBP5 as the research target for reverse validation.

[0073] 1. Tissue acquisition and processing: According to the method in Example 1, 30 live specimens of the middle layer of the AAA artery wall and 40 normal infrarenal aortic control tissues were collected.

[0074] 2. RNA extraction

[0075] RNA extraction was carried out according to the method of Example 1.

[0076] 3. Reverse transcription

[0077] 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. In...

Embodiment 3

[0088] Embodiment 3 interferes with GBP5 gene expression

[0089] 1. Design and synthesis of interference RNA

[0090] According to the GBP5 gene sequence, siRNA was designed and synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd. Shanghai Gemma Pharmaceutical Technology Co., Ltd. also provides a negative control siRNA (siRNA-NC) that has no sequence homology with the GBP5 gene.

[0091] siRNA-GBP5:

[0092] The sense strand is 5'-AACUAAUGUGUGAUUUGGCCA-3' (SEQ ID NO.5);

[0093] The antisense strand is 5'-GCCAAAUCACACAUUAGUUCU-3' (SEQ ID NO.6),

[0094] 2. Culture of human aortic smooth muscle cells

[0095] T / G HA-VSMC cells (referred to as HVSMC cells) were treated with DMEM high-glucose medium containing 10% fetal bovine serum (FBS) plus penicillin 100 units / ml and streptomycin 100 μg / ml, placed at 37°C, 5% CO 2 Cultured in an incubator, the culture medium was changed every 24 hours, and the cells were passaged once every 48 hours. Cells in the logarithm...

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Abstract

The invention discloses application of a GBP5 gene and an expression product thereof as molecular markers for diagnosis of an abdominal aortic aneurysm. Whether a subject suffers from the abdominal aortic aneurysm or has the risk of suffering from the abdominal aortic aneurysm is determined by detecting the contents of the GBP5 gene and the expression product thereof in the abdominal aortic aneurysm tissue of the subject. The results of research on the apoptosis of smooth muscle cells cultured in vitro show that cell apoptosis can be inhibited via interference in the expression of the GBP5 gene. Such research results prove that the GBP5 gene and the expression product thereof are potential drug targets for treatment of the abdominal aortic aneurysm.

Description

technical field [0001] The present invention relates to the field of tumor diagnosis and treatment, more specifically, the present invention relates to a tumor diagnosis method by means of detecting GBP5 abnormality; and a tumor therapeutic agent for inhibiting GBP5 gene or protein. Background technique [0002] Abdominal aortic aneurysm (AAA) refers to a disease characterized by localized expansion and bulging of the abdominal aorta due to lesion or loss of the abdominal aortic wall, with a pulsating mass as the main symptom. AAA is usually defined as the continuous expansion of the three-layer structure of the abdominal aortic arterial wall to more than 1.5 times the diameter of the abdominal aorta at the renal artery. It is a common arterial degenerative disease with a high mortality rate. At present, the specific pathogenesis of abdominal aortic aneurysm is not completely clear. Existing reports have shown that its pathogenesis is closely related to factors such as genet...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574A61K45/00A61K48/00A61K31/713A61P35/00
CPCA61K31/713A61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/57484G01N2333/47
Inventor 李曙光孙锦云
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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