A method and kit for monitoring vascular remodeling phenomenon
A vascular remodeling and phenomenon technology, applied in the direction of material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of heavy occlusion of transplanted blood vessels, transplant failure, economic loss, etc., and achieve the effect of effective pathophysiological changes
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Embodiment 1
[0043] Example 1: Mouse common carotid artery pressure-induced vein graft model experiment
[0044] 1. Materials:
[0045] Microhemostats, vascular clips, polyethylene sleeves, ophthalmic forceps, ophthalmic scissors.
[0046] 2. Method:
[0047] After the surgical instruments were sterilized, the mice were anesthetized (intraperitoneal injection of 10 g / L pentobarbital sodium 50 mg / kg), and atropine sulfate 1.7 mg / kg intramuscularly (3.4 μL / g) was given at the same time to keep the airway smooth. The entire surgical procedure was performed under a stereomicroscope. Take the supine position, expose the neck and fix it on the operating table. A longitudinal incision was made from the middle of the mouse neck from the level of the mandible to the top of the sternum, the right sternomastoid muscle was cut off, and the right common carotid artery was separated as long as possible. cut open. The broken end passed through the polyethylene cannula, the blood vessel and the cannu...
Embodiment 2
[0052] Example 2: In vitro stimulus induction model experiment after cell culture
[0053] 1. Materials
[0054] Culture bottle, pipette, centrifuge tube, waste liquid tank, 75% alcohol cotton ball, alcohol lamp, cultured cells, culture medium (DMEM), fetal bovine serum, 0.125% trypsin, PBS solution, CO 2 Incubator, microscope, clean bench.
[0055] 2. Method
[0056] The cells were subcultured into the pull plate by enzyme digestion method. After the cells grew to 70% density, the cells were starved for 24 hours with serum-free DMEM medium, and after being stimulated by mechanical force (SS) for 1 hour, the culture was continued for 23 hours. The cells were harvested for fluorescent staining.
[0057] The staining method includes the following steps:
[0058] A. The vascular smooth muscle cells cultured in vitro are treated with factors to prepare cell slides;
[0059] B. Wash with washing solution (for example, PBS buffer) (for example, wash at least 3 times for 5 minute...
Embodiment 3
[0069] Example 3: Preparation of the kit for detecting vascular remodeling phenomenon according to the present invention
[0070] Based on the method for monitoring the phenomenon of vascular remodeling established in the experiments of Example 1 and Example 2, the inventor has developed a related kit for detecting the phenomenon of vascular remodeling. The composition of the kit is shown in the following table:
[0071]
[0072] This kit adopts terminal deoxynucleotidyl transferase method, which can incompletely combine with double-stranded DNA fragments broken in cells, so as to detect apoptotic cells.
[0073] This kit uses the nuclear proliferation antigen Ki67 as the detection index for detecting cell proliferation. This is because: the cell cycle can be analyzed by detecting DNA content and by detecting and correlating cell cycle-specific intracellular targets, Ki67 is expressed in G1, S, G2 and M phases, but not in G0 phase.
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