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Application of miR-101b-3p in diagnosis and adjuvant treatment of angiostrongylus cantonensis

A technology of 1. mir-101b-3p, Angiostrongylus, applied in medical preparations containing active ingredients, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problem of no miRNA diagnosis and treatment of Guangzhou Angioplasty nematode infections, etc.

Active Publication Date: 2019-04-02
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no research reports on the diagnosis and treatment of Angiostrongylus cantonensis infection by miRNA

Method used

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  • Application of miR-101b-3p in diagnosis and adjuvant treatment of angiostrongylus cantonensis
  • Application of miR-101b-3p in diagnosis and adjuvant treatment of angiostrongylus cantonensis
  • Application of miR-101b-3p in diagnosis and adjuvant treatment of angiostrongylus cantonensis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Obtaining of Angiostrongylus cantonensis body Acsod3

[0042] 1. Experimental materials

[0043] L3-stage larvae of Angiostrongylus cantonensis were obtained from Bacteria glabrata infected with L1-stage larvae for 21 days. 21 and 28 days after infection, the worms were taken out from the brain tissue of mice and rats, and adult worms were taken out from the lungs of rats at the same time 28 days after infection.

[0044] Kit used in the experiment: Premix Taq TM (TaKaRa Taq TM Version 2.0), TIRzol (Invitrogen), RevertAid First Strand cDNA Synthesis Kit (Thermo Scientific).

[0045] Design and synthesize specific Real-time PCR primers, β-actin as an internal reference control, the specific sequence is as follows:

[0046]

[0047] 2. Experimental method

[0048] The specific operation steps of total RNA extraction of Angiostrongylus cantonensis are as follows:

[0049] The sample was crushed by magnetic beads at 4°C. After adding 200 μl TRIzol to the ...

Embodiment 2

[0056] Example 2 Expression characteristics of Acsod3 in Angiostrongylus cantonensis from different host sources

[0057] 1. Experimental materials

[0058] L3-stage larvae of Angiostrongylus cantonensis were obtained from Bacteria glabrata infected with L1-stage larvae for 21 days. 21 and 28 days after infection, the worms were taken out from the brain tissue of mice and rats, and adult worms were taken out from the lungs of rats at the same time 28 days after infection.

[0059] Mouse anti-mouse β-actin monoclonal antibody, goat anti-mouse fluorescent secondary antibody, goat anti-rat fluorescent secondary antibody (Abcam Company), ECL chemiluminescent colorimetric detection of labeled horseradish peroxidase (HRP) antibody and its associated antigen (Millipore).

[0060] 2. Experimental method

[0061] The specific operation steps for detecting the expression level of AcSOD3 by Western blotting are as follows:

[0062] (1) The sample was crushed with magnetic beads at 4°...

Embodiment 3

[0067] Example 3 Antioxidative protective effect of recombinantly expressed AcSOD3 protein

[0068] 1. Experimental materials

[0069] Hydrogen peroxide, AcSOD3 recombinant expression protein treatment worm body, culture plate, DMEM and penicillin (100IU / ml), amphotericin B (0.25μg / ml) and streptomycin (100μg / ml), 5% carbon dioxide incubator.

[0070] 2. Experimental method

[0071] Under in vitro conditions, the antioxidant protective effect of recombinantly expressed AcSOD3 protein on hydrogen peroxide-treated larvae was detected. The specific operation steps are:

[0072] 1. The PCR product recovered by gel electrophoresis purification and the plasmid vector pGEX-4T-1 were digested with restriction endonucleases SmaI and XhoI. The reaction system was as follows: 4 tubes

[0073]

[0074] 37°C, enzyme digestion reaction time 30min. One version of PCR product and one version of plasmid product were detected by 1% agarose gel electrophoresis, and purified with Takara co...

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Abstract

The invention discloses an application of miR-101b-3p in diagnosis and adjuvant treatment of angiostrongylus cantonensis. The application claims the use of miR-101b-3p or its mimetic or its activatoras a marker or a target gene for gene therapy or drug screening of angiostrongylus cantonensis. According to the invention, miR-101b-3p is involved in the regulation of differential expression of AcSOD3 protein in rats and mice, so that the resistance of the worm-against-host derived reactive oxygen can be regulated, the different antioxidant capacities of the worms in suitable and unsuitable hosts can be regulated, and the parasitic adaptation in suitable and unsuitable hosts can be regulated. Therefore, the expression level of miR-101b-3p can be an important diagnostic indicator and a therapeutic target for the diagnosis of unsuitable host infected with angiostrongylus cantonensis, the application reveals a key mechanism for the survival of angiostrongylus cantonensis in the final host,and provides a new idea for improving and optimizing the existing strategies for prevention and control of angiostrongylus cantonensis.

Description

technical field [0001] The present invention relates to the technical field of diagnosis and treatment of Angiostrongylus cantonensis, and more specifically relates to the application of miR-101b-3p in the diagnosis and auxiliary treatment of Angiostrongylus cantonensis. Background technique [0002] Angiostrongylus cantonensis is one of the important food-borne zoonotic parasites in China. In 1934, Professor Chen Xintao of Sun Yat-sen University first discovered and named the pathogen in the pulmonary artery of Rattus norvegicus captured in the suburbs of Guangzhou. Rats are suitable hosts, while humans and mice are unsuitable hosts. Angiostrongylias cantonensis is an infection caused by eating raw or undercooked snail meat containing larvae of Angiostrongylus cantonensis or food contaminated with mollusk secretions. The larvae often invade the human central nervous system (CNS) causing eosinophilic (≥10%) meningoencephalitis or meningitis. It mainly occurs in tropical re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888A61K31/713A61P33/10
CPCA61K31/713A61P33/10C12Q1/6888C12Q2600/178
Inventor 元冬娟吴忠道罗诗琪曾幸达许莲
Owner SUN YAT SEN UNIV
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