Target spot for diagnosing and treating intracranial aneurysm and application of target spot
An intracranial aneurysm and preparation technology, which is applied in the field of tumor markers, can solve the problem of no research revealing the relationship between intracranial aneurysms, etc., and achieve the effects of good application prospects, strong selectivity, and rapid and accurate quantification.
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Embodiment 1
[0034] The collection of embodiment 1 case
[0035] All tissue samples came to the neurosurgery department of the hospital from October 2012 to May 2014 in patients with intracranial aneurysms diagnosed by DSA, 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
[0036] Example 2 extracts the total RNA in the sample
[0037] 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;
[0038] 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;
[0039] 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;
[0040] 4) 2-8°C, centrifuge at lower than 12000g / min for 10min, discard the supernatant, add 1ml of 75% ethanol, centrifuge at lower than 7500g / min for 5min, discard the supernatant, and spot off;
[0041] 5) After the precipitate is dried (it is translucent, do not completely dry, otherwise the stability will be reduced), dissolve it with RNase-freewater at 55-60°C, and freeze it at -70°C.
Embodiment 3
[0042] Example 3 High-throughput sequencing and analysis
[0043] After RNA extraction, agarose gel electrophoresis is performed. From the results of electrophoresis, 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 the HiSeq2500 high-throughput sequencing platform of Illumina Company for high-throughput transcriptome deep sequencing. The results of data analysis provided by the sequencing company were combined with the literature to screen the differentially expressed gene HAPLN1.
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