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Application of SPHAR (S-phase response) as diagnosis and treatment target for abdominal aortic aneurysm

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

Active Publication Date: 2017-09-19
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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  • Application of SPHAR (S-phase response) as diagnosis and treatment target for abdominal aortic aneurysm
  • Application of SPHAR (S-phase response) as diagnosis and treatment target for abdominal aortic aneurysm
  • Application of SPHAR (S-phase response) as diagnosis and treatment target for abdominal aortic aneurysm

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Gene Chip Screening for Differentially Expressed Genes

[0061] 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.

[0062] 2. RNA extraction and cDNA synthesis

[0063] 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

[0072] Example 2 Verification of Differentially Expressed Genes in Large Samples

[0073] The expression selection gene chip in abdominal aortic aneurysm tissue was selected as the research target to carry out reverse verification.

[0074] 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.

[0075] 2. RNA extraction

[0076] RNA extraction was carried out according to the method of Example 1.

[0077] 3. Reverse transcription

[0078] 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 cen...

Embodiment 3

[0090] Example 3 overexpression of SPHAR gene expression

[0091] 1. SPHAR gene recombinant plasmid construction

[0092] (1) amplifying the coding sequence of the SPHAR gene;

[0093] (2) Design amplification primers;

[0094] (3) The amplified SPHAR gene was connected to the expression vector pcDNA3.0 to construct the pcDNA3.0-SPHAR recombinant expression vector.

[0095] 2. Culture of human aortic smooth muscle cells

[0096] 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 logarithmic growth phase were used for subsequent experiments.

[0097] 3. Cell transfection

[0098] Lipofectamine2000 was used as transfection reagent. The experiment was divided into two groups: negativ...

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Abstract

The invention discloses application of SPHAR (S-phase response) as diagnosis and treatment marker for abdominal aortic aneurysm. SPHAR may be applied to the development of products for diagnosing abdominal aortic aneurysm and drugs for treating abdominal aortic aneurysm. From research findings herein, theoretical basis can be provided for clinicians to make personalized treatment schemes, and novel drug targets can be provided for the development of abdominal aortic aneurysm drugs.

Description

technical field [0001] The invention relates to the field of tumor diagnosis and treatment, more specifically, the invention relates to a tumor diagnosis method by means of detecting SPHAR abnormality; and a tumor therapeutic agent for activating SPHAR 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 genetic factors, in...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57407G01N33/68
Inventor 李曙光孙锦云
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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