阅读说明：本技术 8种代谢产物作为精子受损的生物标记 (8 metabolites as biomarkers of sperm damage ) 是由 张欣宗 唐运革 刘武江 邓存灿 于 2020-07-29 设计创作，主要内容包括：本发明公开了8种代谢产物作为精子受损的生物标记,包括如下步骤：(一)、实验分组：将取样标本分成三份,分别为：A、正常捐精自愿者的精液；B、精索静脉曲张病人手术前精液；C、精索静脉曲张病人手术后精液,(二)、实验步骤：A、将代谢物定性与(相对)定量结果进行是否去噪或筛选的操作,当需要对其进行去噪或筛选的操作,可对其进行先重名物资筛选,然后再对其进行噪音剔除。通过采用本发明设计的检测方法,新发现的8种代谢产物具有AUC>90%的预测效能,通过利用这8种在精液里存在的代谢产物,可以判定精子是否受到损伤以及精液质量的衡量,给精液质量的检测工作带来了极大的便利。(The invention discloses 8 metabolites as biomarkers for sperm damage, which comprises the following steps: (I) experimental grouping: dividing the sampling specimen into three parts, namely: A. semen from a normal donated sperm volunteer; B. seminal fluid before operation for patients with varicocele; C. spermatic varicocele patient postoperative semen, (II) and experiment steps: A. and (3) carrying out denoising or screening operation on the qualitative and (relative) quantitative results of the metabolites, and when the denoising or screening operation is required, firstly screening the duplicate materials and then carrying out noise elimination on the duplicate materials. By adopting the detection method designed by the invention, the newly found 8 metabolites have the prediction efficiency of AUC >90%, and whether the sperm is damaged or not and the semen quality can be judged by utilizing the 8 metabolites existing in the semen, so that great convenience is brought to the semen quality detection work.)

1.8 metabolites as biomarkers of sperm damage characterized by: the method comprises the following steps:

(I) experimental grouping:

dividing the sampling specimen into three parts, namely:

A. semen from a normal donated sperm volunteer;

B. seminal fluid before operation for patients with varicocele;

C. postoperative semen of patients with varicocele;

(II) experiment steps:

A. carrying out denoising or screening operation on the qualitative and (relative) quantitative results of the metabolites, when the denoising or screening operation is required, firstly screening duplicate materials, then carrying out noise elimination on the qualitative and (relative) quantitative results of the metabolites, carrying out normalization on the qualitative and (relative) quantitative results of the metabolites subjected to the denoising or screening operation and the qualitative and (relative) quantitative results of the metabolites not subjected to the denoising or screening operation, and then carrying out standardization to obtain quantitative information for statistical analysis;

B. carrying out univariate statistical analysis and multivariate statistical analysis on the quantitative information for statistical analysis obtained in the step, and then obtaining significant difference metabolites;

C. carrying out high-level data analysis and selectable data analysis on the significant difference metabolites obtained in the steps;

(III) experimental results:

（1）pyrrolidonecarboxylic acid

（2）cabergoline

（3）4-Hydroxy-2-butenoic acid

（4）gamma-lactone

（5）dimethyl dialkyl ammonium chloride

（6）L-Acetylcarnitine

（7）Na-Hexanoyl-Nb-inosityltryptophan

（8）5-L-Glutamyl-taurine

the combined detection of the above 8 substances can diagnose varicocele, or the marker of sperm quality or sperm damage.

2. The 8 metabolites as biomarkers of sperm damage according to claim 1, wherein: and the univariate statistical analysis in the second step comprises Student's t test (Student's t-test) and the like.

3. The 8 metabolites as biomarkers of sperm damage according to claim 1, wherein: the multivariate statistical Analysis in the second step includes Principal Component Analysis (PCA), Orthogonal partial least squares-Discriminant Analysis (OPLS-DA), and the like.

4. The 8 metabolites as biomarkers of sperm damage according to claim 1, wherein: and the high-level data analysis in the second step comprises correlation analysis, ROC curve and the like.

5. The 8 metabolites as biomarkers of sperm damage according to claim 1, wherein: the selectable data Analysis in the second step comprises hierarchical clustering Analysis of the differential metabolites, radar map Analysis of the differential metabolites, correlation Analysis of the differential metabolites, KEGG annotation of the differential metabolites, metabolic Pathway Analysis (Pathway Analysis) of the differential metabolites and regulatory Network Analysis (Network Analysis) of the differential metabolites.

Technical Field

The invention relates to the technical field of infertility, in particular to 8 metabolites as biomarkers for sperm damage.

Background

In the early stage, 8 kinds of metabolite micromolecules are screened out by utilizing metabonomics technology from comparison among 30 samples of normal people, patients with varicocele and patients with improved sperm quality after the varicocele, the sperm damage marker of the patient with varicocele can be effectively diagnosed by utilizing the 8 kinds of metabolites to detect singly or jointly, the sperm damage marker can be used for diagnosing the varicocele and judging whether an operation pointer exists, and whether the sperm is damaged or not and the sperm quality can be judged by utilizing the 8 kinds of metabolites existing in the sperm.

Disclosure of Invention

The invention aims to provide 8 metabolites as biomarkers for sperm damage, has the advantage of rapid detection, and solves the problems of long detection time and low detection accuracy of the traditional semen detection method.

In order to achieve the purpose, the invention provides the following technical scheme: 8 metabolites as biomarkers of sperm damage comprising the steps of:

(I) experimental grouping:

dividing the sampling specimen into three parts, namely:

A. semen from a normal donated sperm volunteer;

B. seminal fluid before operation for patients with varicocele;

C. seminal fluid after operation of patients with varicocele.

(II) experiment steps:

A. carrying out denoising or screening operation on the qualitative and (relative) quantitative results of the metabolites, when the denoising or screening operation is required, firstly screening duplicate materials, then carrying out noise elimination on the qualitative and (relative) quantitative results of the metabolites, carrying out normalization on the qualitative and (relative) quantitative results of the metabolites subjected to the denoising or screening operation and the qualitative and (relative) quantitative results of the metabolites not subjected to the denoising or screening operation, and then carrying out standardization to obtain quantitative information for statistical analysis;

B. carrying out univariate statistical analysis and multivariate statistical analysis on the quantitative information for statistical analysis obtained in the step, and then obtaining significant difference metabolites;

C. and (3) carrying out high-level data analysis and optional data analysis on the significant difference metabolites obtained in the steps.

(III) experimental results:

（1）pyrrolidonecarboxylic acid

（2）cabergoline

（3）4-Hydroxy-2-butenoic acid

（4）gamma-lactone

（5）dimethyl dialkyl ammonium chloride

（6）L-Acetylcarnitine

（7）Na-Hexanoyl-Nb-inosityltryptophan

（8）5-L-Glutamyl-taurine

the combined detection of the above 8 substances can diagnose varicocele, or the marker of sperm quality or sperm damage.

Preferably, the univariate statistical analysis in the second step comprises Student's t test (Student's t-test) and the like.

Preferably, the multivariate statistical Analysis in the second step includes Principal Component Analysis (PCA), Orthogonal partial least squares-Discriminant Analysis (OPLS-DA), and the like.

Preferably, the advanced data analysis in the second step includes correlation analysis, ROC curve, and the like.

Preferably, the selectable data Analysis in the second step includes hierarchical clustering Analysis of differential metabolites, radar mapping Analysis of differential metabolites, correlation Analysis of differential metabolites, KEGG annotation of differential metabolites, metabolic Pathway Analysis (Pathway Analysis) of differential metabolites, and regulatory Network Analysis (Network Analysis) of differential metabolites.

Compared with the prior art, the invention has the following beneficial effects:

by adopting the detection method designed by the invention, the newly found 8 metabolites have the prediction efficiency of AUC >90%, and whether the sperm is damaged or not and the semen quality can be judged by utilizing the 8 metabolites existing in the semen, so that great convenience is brought to the semen quality detection work.

Drawings

FIG. 1 is a schematic diagram of the experimental procedure of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The invention provides a technical scheme that:

8 metabolites as biomarkers of sperm damage comprising the steps of:

(I) experimental grouping:

dividing the sampling specimen into three parts, namely:

A. semen from a normal donated sperm volunteer;

B. seminal fluid before operation for patients with varicocele;

C. seminal fluid after operation of patients with varicocele.

(II) experiment steps:

A. carrying out denoising or screening operation on the qualitative and (relative) quantitative results of the metabolites, when the denoising or screening operation is required, firstly screening duplicate materials, then carrying out noise elimination on the qualitative and (relative) quantitative results of the metabolites, carrying out normalization on the qualitative and (relative) quantitative results of the metabolites subjected to the denoising or screening operation and the qualitative and (relative) quantitative results of the metabolites not subjected to the denoising or screening operation, and then carrying out standardization to obtain quantitative information for statistical analysis;

B. subjecting the quantitative information for statistical Analysis obtained in the above steps to univariate statistical Analysis including Student's t-test (Student's t-test) and the like and multivariate statistical Analysis including Principal Component Analysis (PCA), Orthogonal partial least squares-Discriminant Analysis (Orthogonal projects to last Structures-Discriminant Analysis, OPLS-DA) and the like, and then obtaining significantly different metabolites;

C. and performing high-level data Analysis and selectable data Analysis on the significant differential metabolites obtained in the steps, wherein the high-level data Analysis comprises correlation Analysis, ROC curve and the like, and the selectable data Analysis comprises hierarchical clustering Analysis of the differential metabolites, radar map Analysis of the differential metabolites, correlation Analysis of the differential metabolites, KEGG annotation of the differential metabolites, metabolic Pathway Analysis (Pathway Analysis) of the differential metabolites and Network Analysis (Network Analysis) of the differential metabolites.

(III) experimental results:

（1）pyrrolidonecarboxylic acid

（2）cabergoline

（3）4-Hydroxy-2-butenoic acid

（4）gamma-lactone

（5）dimethyl dialkyl ammonium chloride

（6）L-Acetylcarnitine

（7）Na-Hexanoyl-Nb-inosityltryptophan

（8）5-L-Glutamyl-taurine

the 8 substances are jointly detected to diagnose varicocele, or the marker of sperm quality or the marker of sperm damage, by adopting the detection method designed by the invention, the newly found 8 metabolites have the prediction efficiency of AUC >90%, and by utilizing the 8 metabolites existing in the semen, whether the sperm is damaged or not and the measurement of the semen quality can be judged, thereby bringing great convenience to the work of detecting the semen quality.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.