Establishment and application of liver cancer prediction model based on differentiated expression miRNA in fucosylated extracellular vesicles
A fucosylation and differentiation technology, applied in the field of biomedicine, can solve the problems that early diagnosis liquid biopsy is still in the early research stage, achieve excellent clinical application performance, avoid limitations, and improve the effect of predictive ability
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment example 1
[0060] Implementation Case 1: Subject Sample Information
[0061] In this implementation case, the subject sample information includes three sample sets, including:
[0062] Retrospective sample collection: 61 samples from patients with liver cancer, 24 samples from patients with other liver diseases other than liver cancer, and 30 samples from healthy people without liver-related diseases;
[0063] RT-PCR verification sample collection: 16 samples from patients with liver cancer, 16 samples from patients with other liver diseases other than liver cancer, and 16 samples from healthy people without liver-related diseases;
[0064] Prospective sample collection: 52 samples from patients with liver cancer, 30 samples from patients with other liver diseases other than liver cancer, and 23 samples from healthy people without liver-related diseases;
[0065] The above sample collection is used for the subsequent implementation cases of glycosylated extracellular vesicles isolation,...
Embodiment example 2
[0067] Example 2: Isolation and enrichment of fucosylated extracellular vesicles
[0068] In this example, in order to better illustrate the separation and enrichment method and effect of fucosylated extracellular vesicles, conventional differential ultracentrifugation (UC) was used to separate total extracellular vesicles in the sample, and GlyExo- The Capture sugar capture technology isolates and enriches fucosylated extracellular vesicles.
[0069] 1. Use differential ultracentrifugation to separate (UC) total extracellular vesicles from serum samples
[0070] (1) After the serum samples were thawed at 37°C, centrifuge at 3,000 × g for 10 minutes to remove cell debris;
[0071] (2) Dilute the 500 μl serum sample separated in step (1) with 20 times the volume of phosphate buffered saline (PBS), and use a 0.22 μm diameter filter to remove large particles in the serum sample;
[0072] (3) Ultracentrifugation at 100,000 × g for 90 min at 4°C using a 100-type TI rotor with a m...
Embodiment example 3
[0081] Example 3: Identification of extracellular vesicles after isolation
[0082] The total extracellular vesicles isolated by conventional differential ultracentrifugation and the fucosylated extracellular vesicles isolated and enriched by GlyExo-Capture sugar capture technology in Example 2 were analyzed by transmission electron microscopy. , TEM) and nanoparticle tracking analysis (NTA) to evaluate the morphology and size distribution of extracellular vesicles; extracellular vesicles were further verified by Western blot and Exoview analysis techniques. Among them: Transmission Electron Microscopy (TEM) was used to observe the morphology and structure of extracellular vesicles and measure their size; Nanoparticle Tracking Analysis (NTA) was used to measure the diameter distribution and number of EVs particle populations; Western Blot and Exoview analysis techniques identified Marker proteins on the surface of extracellular vesicles to identify whether the isolates are ext...
PUM
| Property | Measurement | Unit |
|---|---|---|
| Particle size | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More