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Method for tracing cervical deep lymph nodes and meningeal lymphatic vessels by using nanometer probes

A nanoprobe and lymph node technology, applied in the field of nanoprobes, can solve the problems of presenting the complete structure of the central nervous system, and achieve the effect of a simple and direct detection method

Active Publication Date: 2018-08-24
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This major discovery fills the gap in the central nervous system, but there is still no definite evidence to present the complete structure of the central nervous system

Method used

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  • Method for tracing cervical deep lymph nodes and meningeal lymphatic vessels by using nanometer probes
  • Method for tracing cervical deep lymph nodes and meningeal lymphatic vessels by using nanometer probes
  • Method for tracing cervical deep lymph nodes and meningeal lymphatic vessels by using nanometer probes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation and characterization of nanoprobes loaded with Cy5

[0027] In this example, by optimizing the ratio of aqueous phase / organic phase and the liquid flow rate of each channel, a nanoprobe with a particle size of 42.4 nm and a surface potential of -21.4 mV was finally prepared. The specific implementation steps are as follows: ① Take 6 mg PLGA, 0.9 mgCy5 was dissolved in 3 mL organic solvent acetonitrile, vortexed and mixed; 15 mg PF127 and 1 mg DSPE-PEG were dissolved in 3 mL ultrapure water, vortexed and mixed; ②Acetonitrile solution containing PLGA and ICG passed through the channel 1 (flow rate 10 mL / min), the aqueous solution containing PF127 and DSPE-PEG passes through channel 2 (flow rate 10 mL / min), 12 mL ultrapure water passes through channels 3 and 4 (flow rate 40 mL / min) respectively, and is delivered to the multi-channel Perform high-speed mixing and solvent exchange in a vortex mixer to obtain unpurified nanoparticles; ③Use a dialysis bag...

Embodiment 2

[0028] Example 2 Real-time imaging of deep cervical lymph nodes and meningeal lymph nodes after injection of nanoprobe CNP

[0029] In this example, the nanoprobe prepared in Example 1 (the concentration of Cy5 is 200 μg / mL) was injected subcutaneously in the right neck of BALB / c mice, and the deep layer of the neck of the mice was surgically exposed Lymph nodes and meningeal lymphatic vessels were observed in real time using an in vivo confocal microscope for changes in their structure and drug concentration. The collective implementation steps are as follows: ① The mice were anesthetized with isoflurane, and 0.2 mL of the nanoprobe CNP was subcutaneously injected near the neck bone of the mouse on the right side of the neck, so that the dose of the probe received by the mice was 2 mg / kg; ② The mice were cut open. Neck skin, blunt dissection of mouse neck musculature, exposing neck deep lymph nodes and nearby meningeal lymphatic vessels; Real-time imaging of deep lymph nodes...

Embodiment 3

[0030] Example 3 Dosage of Nanoprobe CNP in Peripheral Lymph Nodes at Different Time Points

[0031] This example aims to explore the time point when the nanoprobe reaches the highest dose enrichment in the deep cervical lymph nodes. The collective implementation steps are as follows: ①Take 12 mice and divide them into 4 groups randomly, 3 in each group; ②Take one of them The mice in the group were used as the blank control group. After the mice were killed by neck dislocation, a pair of superficial cervical lymph nodes, deep cervical lymph nodes, left axillary lymph nodes, and right axillary lymph nodes were removed from the mice, and the tissue homogenate was broken. The light absorption value of each sample at 650 nm was detected by a functional microplate reader; ③The other three groups of mice were injected with 0.2 mL of CNP, and a group of mice were sacrificed at 12 hours, 24 hours and 48 hours, and the small A pair of superficial cervical lymph nodes, deep cervical lym...

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Abstract

The invention belongs to the technical field of nanometer probes, and in particular relates to a method for tracing cervical deep lymph nodes and meningeal lymphatic vessels by using nanometer probes.According to the method, core-shell nanoparticles prepared by using polyglycolide lactide and pluronic F127 as main raw materials are used as a model nano drug-loading system (with a particle size of20-50nm), and are coated with a near-infrared fluorescent dye Cy5 so as to form the nanometer probes. After being subjected to subcutaneous injection in the right neck, the nanometer probes are enriched to the superficial lymph nodes, deep cervical lymph nodes and axillary lymph nodes by means of local lymphatic drainage of tissues, wherein the deep cervical lymph nodes and the meningeal lymphatic vessels are directly connected with each other. Therefore, the nanometer probes can be rapidly transmitted to the meningeal lymphatic vessels for real-time imaging tracing of the deep cervical lymphnodes and the meningeal lymphatic vessels, thus providing a simple and direct detection method for the lymphatic transport research of the nanoparticles in the neck and the physiological and pathological study of the brain lymphatic system.

Description

technical field [0001] The invention relates to the technical field of nanoprobes, in particular to a method for tracing cervical deep lymph nodes and meningeal lymphatic vessels by using nanoprobes. Background technique [0002] The lymphatic system is another set of liquid reticular circulatory system independent of the blood circulatory system, including lymphatic tissue, lymphatic vessels and lymph fluid. meaning. Studies have shown that the lymphatic system has an important impact on the occurrence and development of diseases. For example, malignant tumors often infiltrate into the lymph nodes surrounding the primary site before metastasizing to the tissue. Therefore, it is particularly important to prevent and treat diseases by developing and imaging the lymphatic system to study the structure and physiological functions of the lymphatic system. [0003] The research on the lymphatic system of the human body has been relatively thorough, but the research on the lymp...

Claims

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Application Information

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IPC IPC(8): A61K49/00
CPCA61K49/0032A61K49/0034A61K49/0054A61K49/0067A61K49/0093
Inventor 赵鹏飞陈永明刘立新
Owner SUN YAT SEN UNIV
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