New application of csf2 gene and its expression product
A technology of gene and gene suppression, applied in the application field of CSF2 gene and its expression products in the diagnosis and treatment of intracranial aneurysms, can solve the problems of interfering with CSF2 gene, no research revealing the relationship between intracranial aneurysms, and affecting the activity of EPCs, etc., to achieve Fast and accurate quantification, good application prospects, and strong selectivity
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Embodiment 1
[0035] The collection of embodiment 1 case
[0036] All tissue samples were from patients with intracranial aneurysms diagnosed by DSA from October 2012 to May 2014, and underwent microsurgical craniotomy. The aneurysm tissue samples were the aneurysm wall tissue removed during microsurgery, and the control normal cerebrovascular tissue samples were the superficial temporal artery (STA) and / or middle meningeal artery (MMA) of brain tumor craniotomy patients at the same period. In order to minimize the differences between samples, all aneurysm tissue samples were selected from saccular aneurysms. The obtained tissue samples were immediately placed in a liquid nitrogen tank.
Embodiment 2
[0037] Example 2 High-throughput sequencing and analysis
[0038] After the RNA is extracted, agarose gel electrophoresis is performed. From the electrophoresis results, it can be preliminarily judged whether the quality of the extracted RNA sample is qualified or not, and whether it can be used for further transcriptome sequencing. Then, the extraction of RNA samples was detected by NanoDrop1000 spectrophotometer, and the sample requirements for RNA-seq sequencing: OD260 / OD280 was 1.8-2.2. The qualified samples were sent to the sequencing company for sequencing. The sequencing platform was HiSeq 2500 high-throughput sequencing platform of Illumina Company for deep sequencing of high-throughput transcriptome. The results of data analysis provided by the sequencing company were combined with the literature to screen the differentially expressed gene CSF2.
Embodiment 4
[0039] Example 4 CSF2 gene expression of intracranial aneurysm patient group and control group
[0040] 1. Materials
[0041] Aneurysm wall tissues from 23 cases of intracranial aneurysm patients and 11 cases of normal control cerebrovascular samples were selected and grouped and numbered. See Example 1 for sample collection criteria.
[0042] 2. Method
[0043] 2.1 Extraction of RNA
[0044] 1) Put small pieces of aneurysm (normal control blood vessel) tissue in a petri dish, add 1ml Trizol, cut it into pieces with ophthalmic scissors, and transfer to a homogenizer for homogenization;
[0045] 2) After the homogenization is completed, take out the homogenate and place it in an EP tube, set the volume to 1ml, and store it at -70°C;
[0046] 3) Place at 15-30°C for 5 minutes, add 200 μl chloroform, shake vigorously for 15 sec, and place at 15-30°C for 10 minutes;
[0047] 4) 2-8°C, centrifuge at less than 12000g / min for 10min, discard the supernatant, add 1ml of 75% ethano...
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