Lipid biomarker for autism and application of lipid biomarker

A biomarker and autism technology, applied in the field of disease diagnosis, can solve the problems of few literature reports

Pending Publication Date: 2021-09-28
THE SEVENTH AFFILIATED HOSPITAL SUN YAT SEN UNIV SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, little has been reported in the literature

Method used

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  • Lipid biomarker for autism and application of lipid biomarker
  • Lipid biomarker for autism and application of lipid biomarker
  • Lipid biomarker for autism and application of lipid biomarker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 16p11.2 + / - Microstructural changes of myelin sheath in mouse striatum

[0023] Mice were anesthetized with 0.1 g / ml chloral hydrate and the heart was perfused first with 20 ml of 1X Phosphate Buffered Saline (PBS) followed by 60 ml of fixative (4% PFA + 2.5% glutaraldehyde in 1X PBS) . Brains were removed and striatum were harvested using brain molds, which were then transferred to EP tubes filled with fresh TEM fixative (Servicebio) for further fixation, stored and shipped at 4°C.

[0024] Tissues were washed 3 times with 0.1 M PB (pH 7.4) for 15 minutes each, and then fixed in 1% OsO4 in 0.1 M PB (pH 7.4) for 2 hours at room temperature protected from light. After removal of OsO4, the tissue was rinsed three times for 15 minutes each in 0.1 M PB (pH 7.4). Dehydrate the tissue at room temperature as follows: 30% ethanol for 20 min; 50% ethanol for 20 min; 70% ethanol for 20 min; 80% ethanol for 20 min; 95% ethanol for 20 min; two changes of 100% ethanol f...

Embodiment 2

[0027] Example 2 Myelin-related genes at 16p11.2 + / - Expression changes in mouse striatum

[0028] (1) RNA sequencing

[0029] Four wild-type (WT) female mice and four 16p11.2 mice were used at postnatal day 60 + / - female mice. The mice were terminally anesthetized with 0.1 g / ml chloral hydrate (Macklin, Shanghai), and the striatal tissue was excised and immediately frozen in liquid nitrogen. RNA extraction and RNA sequencing (RNAseq) analysis were performed by Applied Protein Technology Co. Briefly, total RNA was isolated using TRIzol RNA Isolation Reagent and treated with RNase-free DNase I to remove genomic DNA. A total of 2 μg RNA per sample was used as input material for RNA sample preparation. A cDNA library (paired-end 250bp, PE250) was constructed, and HiSeq 2000 sequencing system was used (high-throughput sequencing was performed). After filtering out low-quality sequences and adapter sequences, reads were mapped to the mouse genome using Hisat2 software (versio...

Embodiment 3

[0035] Example 3 16p11.2 + / - Altered lipidomic profiles in the mouse striatum

[0036] Six WT female mice and five 16p11.2 mice were used at P60 + / - female mice. Mice were terminally anesthetized with 0.1 g / ml chloral hydrate, striatal tissue was excised and immediately frozen in liquid nitrogen. Lipid extraction and mass spectrometry-based lipid detection were performed by Applied Protein Technology Co. A small fraction of samples from each group (WT and 16p11.2 + / - ) to create pooled QC samples. Insert quality control samples into the analysis queue to assess system stability and data reliability throughout the experiment. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) was performed on a Q Exactive plus mass spectrometer (Thermo Fisher Scientific) combined with UHPLCNexera LC-30A (Shimadzu Co. Beijing). Lipid identification, peak extraction, peak alignment and quantification were evaluated using LipidSearch software version 4.1 (Thermo Fisher Scientific).

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Abstract

The invention relates to the field of disease diagnosis, in particular to a lipid biomarker for autism and application of the lipid biomarker. The invention discloses the effect of myelination in autism, and the result that reduction of myelination is an important reason of onset of autism possibly is found. Sphingomyelin (SM) and hexosyl ceramide (CerG1) are two types of important lipids forming a myelin sheath; and lipiomics results show that the contents of sphingomyelin and hexosyl ceramide of long acyl chains in autism model mice are significantly reduced. The two lipids can be used as potential biomarkers for autism diagnosis, and have important clinical prospects and economic values.

Description

technical field [0001] The invention relates to the field of disease diagnosis, in particular to a lipid biomarker for autism and its application. Background technique [0002] Autism spectrum disorders (ASD) are developmental disorders characterized by difficulties communicating and interacting with others, limited interests, repetitive behaviors, and intellectual disability. It is estimated that the prevalence of ASD in the world is 0.76%, while the prevalence of ASD in my country is about 1.28%. Although the exact cause of ASD is unknown, a combination of genetic variation and environmental factors has been identified as a risk factor for ASD. Among them, the deletion of a region with a length of about 600k base pairs in the 16p11.2 locus is one of the most common phenomena in ASD, with an incidence rate of about 1 / 2000. [0003] Interestingly, evidence suggests a strong correlation between the pathogenesis of autism and abnormalities in lipid metabolism due to genetic ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883G01N33/92
CPCC12Q1/6883G01N33/92C12Q2600/158G01N2800/30
Inventor 李宁宁鞠俊朱文辉
Owner THE SEVENTH AFFILIATED HOSPITAL SUN YAT SEN UNIV SHENZHEN
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