Biomarker for ICH prognosis evaluation and application thereof
An exosome and plasma technology, applied in the field of medical biology, can solve the problems of inability to meet clinical needs and the high value of ICH prognosis prediction, and achieve the effect of improving clinical services.
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Embodiment 1
[0065] Example 1: Clinical data collection of ICH patients and health control group
[0066] This study was included in the patients who were hospitalized in Beijing Tiantan Hospital affiliated to Capital Medical University from October 2018 to the Patients of Patients, Beijing Tiantan Hospital, November 2019. ICH patient inclusion standard: (1) The age of onset is between 18-80; (2) According to the patient's clinical and imaging data, it is clearly diagnosed with acute cerebral hemorrhage; (3) This disease is due to hypertensiveness; ) The bleeding position is around the foundation, and the bleeding is not broken into the brain, the bleeding volume is less than 50 ml. The clinical information includes: patients generally basic information and clinical characteristics, high risk of martial arterygene, high blood pressure, diabetes, coronary heart disease, high blood fat, etc.), acute period treatment plan. Follow-up information includes: March, June 1, NIHSS and MRS scores, recu...
Embodiment 2
[0067] Example 2: Sample Collection and Storage of ICH Patients and Health Control Group
[0068] 2.1 Plasma Collection - X Peripheral Blood is collected in a purple tube containing anti-condensing agent EDTA or heparin, centrifuged in 30 min after sample collection, 3000 rpm 10min, 2-8 ° C. The upper plasma was collected, stored in batches to -80 ° C. Sample avoids repeated freeze-thaw. Note: The sample should be sufficiently centrifuged to avoid hemolysis or particle presence.
[0069] 2.2 Extraction of Sino-Foreout in Peripheral Plasma
[0070] We use serum and plasma EXOQUICKOBOFO Separation kits (item numbers: Equltra-20A-1), an innovative kit with expertise in exosomes.
[0071] 2.2.1 extraction
[0072] 1) Remove the sample, 3000g, and 15 min from centrifugation.
[0073] 2) If there is still a cell debris residue, the supernatant is once again at a position 12,000g, 10 min, and the supernatant is transferred to a new centrifuge tube.
[0074] 3) After obtaining 250 μL sam...
Embodiment 3
[0100] Example 3: Gene, proteolytic technology analysis, screening of differentially expressing proteins, resulting in recurrence, prognosis Detection combination of material tag
[0101] 3.1 This project uses the next generation of non-marked quantitative proteomics techniques to complete analysis, in Data Independent Acquisition, DIA mode, it provides unparalleled protein group coverage while achieving exact amount of protein per sample , Highly repeatable quantification. The DIA process provides an ideal differentially expressing protein group qualitative analysis or a protein set quantification platform of massive samples. The DIA process is based on three necessary steps:
[0102] 1) Construction Spectral Stock: Spectral Library Collect all detectable non-redundant high quality peptide information (MS / MS spectrum), as a peptide identification template for subsequent data analysis. It includes a fragment ion intensity and retention time describing the peptide peak characte...
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