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Screening method and use of biomarker related to severe oligoasthenospermia

A biomarker, asthenozoospermia technology, used in instruments, measuring devices, scientific instruments, etc.

Active Publication Date: 2017-06-20
SHANDONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports on non-coding amino acids as biomarkers associated with severe oligoasthenospermia

Method used

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  • Screening method and use of biomarker related to severe oligoasthenospermia
  • Screening method and use of biomarker related to severe oligoasthenospermia
  • Screening method and use of biomarker related to severe oligoasthenospermia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] Example 1: Screening of biomarkers associated with severe oligoasthenospermia

[0125] The specific screening method is as follows:

[0126] 1. Sample processing and experimental analysis

[0127] 1. Extraction of the whole protein of sperm cells: the same amount of severe oligospermia and normal sperm samples were washed three times with DPBS, and the same amount of RIPA lysate was added to sonicate for 1-2 minutes, incubated on ice for 30 minutes to lyse, and centrifuged at 4°C at 14,000g ×20min to take the supernatant. Protein concentration was determined using the Bradford method.

[0128]2. Proteolysis: Take approximately 150 μg of sperm protein from the samples of severe oligospermia and normal sperm, use 10% polyacrylamide gel electrophoresis (SDS-PAGE) to separate the proteins, and divide each into 5 parts for enzymatic hydrolysis. Peptides were desalted using ziptip.

[0129] 3. Mass Spectrometry: Nanoflow Liquid Chromatography Separation: Phase A: Water co...

Embodiment 2

[0185] Embodiment 2: clinical detection verification

[0186] Taking 4 healthy samples and 8 clinically diagnosed severe oligoasthenospermia samples as the research objects for verification, the serine at the 208th position of the AKAP3 protein and the mass deviation of -113.05347 at the 186th position of the AKAP4 protein were detected respectively in the above samples. Asparagine shifted, asparagine with mass shift of -114.04278 at position 186 of AKAP4 protein, glutamine with mass shift of -17.02660 at position 617 of AKAP4 protein, lysine with mass shift of +211.09682 at position 733 of AKAP4 protein , lysine with +42.01108 mass shift at position 531 of ATP5A1 protein, lysine with +42.01108 mass shift at position 87 of COX4I1 protein, threonine with +79.96685 mass shift at position 64 of GAPDHS protein and 692 position of KIAA1683 protein The respective detection frequencies of serine with +79.96685 mass shift occurred, and the samples to be tested were diagnosed according...

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Abstract

The invention discloses a method for screening a biomarker related to severe oligoasthenospermia. The method comprises carrying out deep mass spectrometry on multiple sperm protein uncoded amino acids of severe oligoasthenospermia through a NanoHPLC-MS / MS mass spectrometry system and an unlabeled quantitative proteomics method, searching the mass spectrometry data through a nonfinite amino acid protein modification analysis method, carrying out multivariable gaussian mixture distribution clustering analysis to identify the non-coding amino acids in the sperm protein group as much as possible, and comparing the non-coding amino acids of the normal and patient sperm protein groups to obtain protein uncoded amino acid sites related to severe oligoasthenospermia, wherein the protein uncoded amino acid sites are used as molecular markers of severe oligoasthenospermia. The method provides novel diagnostic and therapeutic targets for severe oligoasthenospermia.

Description

technical field [0001] The invention relates to the technical fields of medicine and molecular diagnosis, in particular to the screening and application of biomarkers related to severe oligoasthenospermia. Background technique [0002] Infertility has become a worldwide reproductive health problem, of which the infertility caused by male factors accounts for about 50%, and it is on the rise in recent years. The main cause of male infertility is oligozoospermia. According to the World Health Organization standard, if the number of a-class sperm is less than 25%, the number of (a+b)-class sperm is less than 50%, and the sperm motility rate is lower than 60%, it can be diagnosed as asthenozoospermia. Oligospermia refers to a condition in which the number of sperm in semen is lower than that of normal fertile men. When a man's sperm is less than 20 million per milliliter, it is oligospermia. [0003] There are many studies on the sperm proteome. Saraswat et al. used UPLC-MS t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8831
Inventor 杨静华陈新骏吕鑫赵涵李翠玲毕白斌巩晶王凤芹孙胜楠王兴元陈子江杨静鸣
Owner SHANDONG UNIV
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