Marker combination for risk assessment when individual is exposed to electromagnetism and / or noise and application thereof

Abstract

The invention belongs to the technical field of molecular diagnosis, and particularly relates to a marker combination for risk assessment when an individual is exposed to electromagnetism and / or noise and application of the marker combination, and markers comprise one or more of 1-stearoyl-2-linoleoyl-sn-glycerin, 1-(9Z, 12Z-octadecadienoyl)-lysophosphatidic acid and N, N-dimethyl phosphatidyl ethanolamine (16: 0 / 16: 0). According to the present invention, the marker can significantly change in the early stage (less than or equal to 7 days) after the individual is exposed by electromagnetism and / or noise, and the abundance level of the marker in the blood sample from the individual is detected so as to rapidly achieve the early, accurate and non-invasive detection of the health risk when the individual is exposed by electromagnetism and / or noise, such that the application prospect is wide.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of molecular diagnosis, and particularly relates to a marker combination for risk assessment when an individual is exposed to electromagnetic and / or noise and application thereof. BACKGROUND

[0002] At present, the assessment of health risks caused by exposure of an individual to electromagnetic and / or noise mainly relies on targeted detection of limited and known traditional serum biomarkers, such as cardiac troponin I (cTn-I), S100 calcium binding protein β (S100-β), etc. However, the traditional serum markers usually change significantly in serum only after obvious pathological damage occurs in tissues, and thus are a kind of “post-event” detection technology, and have problems such as low detection throughput, long cycle, insufficient sensitivity, etc.

[0003] Therefore, it is urgent to develop a marker capable of realizing early monitoring, high sensitivity and non-invasive early warning, which is of great significance for accurately assessing the health risks of an individual exposed to electromagnetic and / or noise, timely intervention and prevention of related diseases. SUMMARY

[0004] The present application aims to at least partially solve at least one of the technical problems existing in the prior art. To this end, the present application provides a marker combination for risk assessment when an individual is exposed to electromagnetic and / or noise and application thereof. The marker of the present application will change significantly in the early stage (≤7 days) after the individual is exposed to electromagnetic and / or noise, and by detecting the abundance level of the marker in a blood sample derived from the individual, the early, accurate and non-invasive detection of health risks of the individual exposed to electromagnetic and / or noise can be realized quickly, and the application prospect is wide.

[0005] In a first aspect of the present application, the present application provides a marker for risk assessment when an individual is exposed to electromagnetic and / or noise. According to an embodiment of the present application, the marker comprises one or more of 1-stearoyl-2-linoleoyl-sn-glycerol, 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid and N,N-dimethylphosphatidylethanolamine (16:0 / 16:0).

[0006] In a second aspect, the present application provides use of a reagent for detecting 1-stearoyl-2-linoleoyl-sn-glycero, a reagent for detecting 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid, or a reagent for detecting N,N-dimethylphosphatidylethanolamine (16:0 / 16:0) in the preparation of a kit. According to embodiments of the present application, the kit is used for risk assessment of an individual exposed to electromagnetic and / or noise.

[0007] In a third aspect, the present application provides a kit for risk assessment of an individual exposed to electromagnetic and / or noise. According to embodiments of the present application, the kit contains a reagent for detecting the marker of the first aspect.

[0008] In a fourth aspect, the present application provides a method for assessing the risk of an individual exposed to electromagnetic and / or noise. According to embodiments of the present application, the method comprises: obtaining a sample to be tested, detecting the abundance level of the marker of the first aspect in the sample to be tested, and assessing the degree of risk of the individual exposed to electromagnetic and / or noise environment based on the abundance level; wherein the method is for non-diagnostic purposes.

[0009] According to embodiments of the present application, the above-mentioned method can have the following additional technical features: According to embodiments of the present application, the sample to be tested is a blood sample.

[0010] In a fifth aspect, the present application provides a system or device. According to embodiments of the present application, the system or device comprises: a data acquisition module for acquiring abundance data of the marker of the first aspect in a sample from an individual; and a risk assessment module for assessing the degree of risk of the individual exposed to electromagnetic and / or noise based on the abundance data.

[0011] Additional aspects and advantages of the present application will be given, partially in the following description, partially become obvious from the following description, or be understood by practice of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0012] The above and / or additional aspects and advantages of the present application will become apparent and be readily understood from the following description, taken in conjunction with the accompanying drawings, in which: Figure 1 The flow chart of the behavioral test experiment in Example 1 of the present application is designed; Figure 2Figure of the behavioral test results of each group of rats in Example 1 of the present application, wherein A is a figure of the total distance statistics of each group of rats in the open field experiment, B is a figure of the average speed statistics of each group of rats in the open field experiment, C is a figure of the time statistics of each group of rats entering the central area in the open field experiment, D is a figure of the recognition index statistics of each group of rats in the new object experiment, E is a figure of the percentage of new arm exploration time of each group of rats in the Y maze experiment, F is a figure of the new arm exploration distance ratio of each group of rats in the Y maze experiment, E0N0 is the false radiation group (group C), E0N1 is the noise exposure group (group N), E1N0 is the electromagnetic exposure group (group E), and E1N1 is the electromagnetic and noise combined exposure group (group EN); Figure 3 Figure of the blood index detection results of each group of rats in Example 1 of the present application, wherein A is a figure of the troponin content level detection results of each group of rats, B is a figure of the lactate dehydrogenase content level detection results of each group of rats, C is a figure of the creatine kinase content level detection results of each group of rats, D is a figure of the angiotensin II content level detection results of each group of rats, E is a figure of the testosterone content level detection results of each group of rats, F is a figure of the growth hormone content level detection results of each group of rats, G is a figure of the S100-β content level detection results of each group of rats, H is a figure of the UCHL1 content level detection results of each group of rats, E0N0 is the false radiation group (group C), E0N1 is the noise exposure group (group N), E1N0 is the electromagnetic exposure group (group E), and E1N1 is the electromagnetic and noise combined exposure group (group EN); Figure 4 Figure of the qualitative and quantitative analysis statistics of serum non-target metabolites in Example 1 of the present application; Figure 5 Figure of the intersection of differential metabolites at different time points of serum samples in Example 1 of the present application; Figure 6 Figure of the KEGG enrichment pathway results of differential metabolites at different time points of serum samples in Example 1 of the present application, wherein A is a KEGG enrichment pathway figure of differential metabolites at 0 d after long-term exposure, B is a KEGG enrichment pathway figure of differential metabolites at 7 d after long-term exposure, C is a KEGG enrichment pathway figure of differential metabolites at 14 d after long-term exposure, and D is a KEGG enrichment pathway figure of differential metabolites at 28 d after long-term exposure; Figure 7 Heat map and trend chart of differential metabolites at 7 d after exposure in Example 1 of the present application; Figure 8 Figure of the correlation analysis results of key metabolites and phenotypes at 7 d after exposure (group EN) in Example 1 of the present application; Figure 9The KEGG pathway result chart in which the metabolites involved in the embodiment 1 of the present application are focused on is shown in the following table, in which A is the KEGG enrichment pathway result chart of the differential metabolites on the 0th day after long-term exposure, B is the KEGG enrichment pathway result chart of the differential metabolites on the 7th day after long-term exposure, C is the KEGG enrichment pathway result chart of the differential metabolites on the 14th day after long-term exposure, and D is the KEGG enrichment pathway result chart of the differential metabolites on the 28th day after long-term exposure. DETAILED DESCRIPTION

[0013] Embodiments of the present application are described in detail below. The embodiments described below are exemplary only and are not to be construed as limiting the present application.

[0014] It should be noted that the terms "first", "second" are used only for descriptive purposes and should not be construed as indicating or implying relative importance or an indicated number of technical features. Thus, the features defined with "first", "second" can explicitly or implicitly include one or more of the features. Further, in the description of the present application, unless otherwise stated, the meaning of "a plurality of" is two or more.

[0015] The endpoints of the ranges and any values disclosed herein are not limited to the precise values stated. The ranges or values should be construed to be roughly about the ranges or values. For ranges, the endpoints are included between the values, the endpoints are included with the individual points, and the individual points are included between the endpoints, as if each value between the endpoints and each value within the range is specifically and individually listed herein.

[0016] In this document, the terms "comprises" or "comprising" are open-ended, that is, they mean including, but not limited to, the indicated features.

[0017] In this document, the terms "optionally", "optional" or "option" generally mean that the subsequently described event or circumstance can or can not occur, and the description includes cases where the event or circumstance occurs, and cases where the event or circumstance does not occur.

[0018] Terms and definitions In this document, the term "electromagnetic and / or noise exposure" refers to the simultaneous or sequential exposure of a living body (such as an experimental animal or a human body) to a specific intensity of electromagnetic field (such as S-band and X-band microwaves used in the embodiments of the present application) and / or a specific sound pressure level of noise environment, similar to the stress conditions commonly encountered in real working environments.

[0019] In the present context, the term "serum metabolic marker" refers to small molecule chemicals (typically < 1500 Da in molecular weight) present in blood samples, which are intermediate or end products of metabolic processes in the body, and the changes in their species and concentrations can sensitively reflect the changes in physiological and pathological states of the body under certain stimuli (such as electromagnetic and / or noise exposure), and in particular, endogenous metabolites related to electromagnetic and / or noise exposure discovered by non-targeted metabolomics technology.

[0020] In the present context, the term "non-targeted metabolomics" refers to a global, hypothesis-driven omics analysis technology, which aims to detect as many small molecule metabolites as possible in biological samples without bias, and then through bioinformatics analysis, to find metabolites with significant differences between groups, rather than only detecting specific metabolites known.

[0021] In the present context, the term "risk assessment" refers to the assessment of the probability and extent of health damage that an individual may suffer due to exposure to certain environmental factors (such as electromagnetic and / or noise) by analyzing the levels of markers.

[0022] In the present context, the term "1-stearoyl-2-linoleoyl-sn-glycerol" as one of the newly discovered markers for risk assessment of individuals exposed to electromagnetic and / or noise, i.e. 1-Stearoyl-2-linoleoyl-sn-glycerol, is a diglyceride molecule with stearic acid (18:0) at the sn-1 position and linoleic acid (18:2(9Z,12Z)) at the sn-2 position, which is an important intermediate in the metabolism and signal transduction of glycerophospholipids.

[0023] In the present context, the term "1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid" as one of the newly discovered markers for risk assessment of individuals exposed to electromagnetic and / or noise, i.e. LPA(18:2(9Z,12Z) / 0:0, is a lysophosphatidic acid containing a linoleoyl group in its structure, which can be involved in the regulation of cell proliferation, migration and inflammation through G protein-coupled receptors.

[0024] In the present context, the term "N,N-dimethylphosphatidylethanolamine (16:0 / 16:0)" as one of the newly discovered markers for risk assessment of individuals exposed to electromagnetic and / or noise, i.e. PE-NMe(16:0 / 16:0), is a product of methylation modification of phosphatidylethanolamine, which plays an important role in cell membrane structure maintenance and signal transduction.

[0025] Marker The present application provides a marker for risk assessment when an individual is exposed to electromagnetic and / or noise. According to an embodiment of the present application, the marker comprises one or more of 1-stearoyl-2-linoleoyl-sn-glycerol, 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid and N,N-dimethylphosphatidylethanolamine (16:0 / 16:0). Compared with traditional protein damage markers (such as cTn-I, S100-β, etc.), the marker according to the embodiment of the present application can respond to electromagnetic and / or noise exposure (≤7 days) earlier and more sensitively, much earlier than when obvious pathological damage occurs in tissues, achieving a true "early warning".

[0026] It should be noted that the marker comprises "one or more", specifically including the following cases: the marker is a single marker (1-stearoyl-2-linoleoyl-sn-glycerol, 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid or N,N-dimethylphosphatidylethanolamine (16:0 / 16:0)); a combination of two markers (1-stearoyl-2-linoleoyl-sn-glycerol and 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid, 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid and N,N-dimethylphosphatidylethanolamine (16:0 / 16:0), 1-stearoyl-2-linoleoyl-sn-glycerol and N,N-dimethylphosphatidylethanolamine (16:0 / 16:0)); a combination of three markers (1-stearoyl-2-linoleoyl-sn-glycerol, 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid and N,N-dimethylphosphatidylethanolamine (16:0 / 16:0)), all of which are within the scope of the present application.

[0027] It should be noted that in the embodiments of the present application, the degree of change of 1-stearoyl-2-linoleoyl-sn-glycerol, 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid and N,N-dimethylphosphatidyl ethanolamine (16:0 / 16:0) in the sham radiation group, the electromagnetic exposure group, the noise exposure group and the electromagnetic and noise combined exposure group is sham radiation group < electromagnetic exposure group < noise exposure group < electromagnetic and noise combined exposure group, and it can be seen that the above markers have certain specificity in indicating different degrees of exposure damage; the difference of the same metabolite between different treatment groups is most significant at 7 days after exposure, which indicates that the above markers have certain sensitivity in indicating different degrees of exposure damage; further, the inventors respectively verify the specificity / high sensitivity of 1-stearoyl-2-linoleoyl-sn-glycerol, 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid and N,N-dimethylphosphatidyl ethanolamine (16:0 / 16:0) as markers when an individual is exposed to electromagnetic and / or noise, and the specific conditions are as follows: by setting various controls and gradient exposure experiments, combining ROC curve analysis and trend test, the specificity and sensitivity of the above markers as markers are verified; thus, it is proved that the markers have high specificity and sensitivity, and meet the basic requirements of the markers for detecting the risk assessment of an individual exposed to electromagnetic and / or noise, and the markers of the present application can be used as detection indicators to quickly and accurately assess the risk level of an individual exposed to electromagnetic and / or noise.

[0028] Use in the preparation of a kit The present application provides the use of a reagent for detecting 1-stearoyl-2-linoleoyl-sn-glycerol, a reagent for detecting 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid or a reagent for detecting N,N-dimethylphosphatidyl ethanolamine (16:0 / 16:0) in the preparation of a kit. According to the embodiments of the present application, the kit is used for risk assessment when an individual is exposed to electromagnetic and / or noise. Thus, by introducing the reagent for detecting the above markers into the kit, early, accurate and efficient risk assessment of an individual exposed to electromagnetic and / or noise is achieved.

[0029] Kit The present application provides a kit for risk assessment when an individual is exposed to electromagnetic and / or noise. According to the embodiments of the present application, the kit contains a reagent for detecting the above markers. Thus, the kit can quickly and accurately assess the risk degree of an individual exposed to electromagnetic and / or noise by detecting the abundance level of the above markers.

[0030] Method The present application provides a method for evaluating the risk of electromagnetic and / or noise exposure of an individual. According to an embodiment of the present application, the method comprises: obtaining a sample to be tested, detecting the abundance level of the aforementioned marker in the sample to be tested, and evaluating the degree of risk of the individual exposed to electromagnetic and / or noise environment based on the abundance level; wherein the method is used for non-diagnostic purposes.

[0031] According to an embodiment of the present application, the sample to be tested is a blood sample.

[0032] It should be noted that the "blood sample" of the present application can be obtained from a healthy individual or a patient or a subject, including but not limited to whole blood samples, plasma samples, serum samples, etc.

[0033] System or device The present application provides a system or device. According to an embodiment of the present application, the system or device comprises: a data acquisition module for acquiring the abundance data of the aforementioned marker in a sample from an individual; and a risk assessment module for evaluating the degree of risk of the individual exposed to electromagnetic and / or noise based on the abundance data. In this way, by detecting the abundance level of the aforementioned marker in the sample to be tested, the degree of risk of the individual exposed to electromagnetic and / or noise can be quickly and accurately evaluated.

[0034] It should be noted that the "system or device" regardless of its specific form, as long as it contains the module for acquiring the abundance data of the aforementioned marker and the module for risk assessment based on the data, and performs the functions of the system or device of the present application, such as remote data analysis and service system based on cloud platform, etc., all belong to the protection scope of the present application.

[0035] The scheme of the present application will be explained below in conjunction with the embodiments. Those skilled in the art will understand that the following embodiments are only used to illustrate the present application, and should not be regarded as limiting the scope of the present application. If the specific technology or condition is not specified in the embodiments, it is carried out according to the technology or condition described in the literature in the art or according to the product instruction. If the reagent or instrument used is not specified by the manufacturer, it is a conventional product that can be obtained by purchase.

[0036] Example 1: Screening and verification of serum metabolic markers of rats exposed to long-term electromagnetic and noise 1. Experimental animals and grouping treatment One hundred and twenty-eight 8-week-old, 250-280 g male Wistar rats (purchased from Beijing Sibeifeng Experimental Animal Technology Co., Ltd.) were randomly divided into four groups: sham exposure group (group C), electromagnetic exposure group (group E), noise exposure group (group N), and electromagnetic and noise combined exposure group (group EN). Each group was set up four detection time points, namely immediately after the end of exposure (0 d), 7 d, 14 d, and 28 d. In addition, according to different detection contents, the rats in each group were divided into rats required for behavior test (a total of 40 rats, 10 rats in each group), rats required for physiological function detection (a total of 48 rats, 3 rats in each group at each time point), rats required for biochemical and reproductive function analysis and pathological evaluation (a total of 80 rats, 5 rats in each group at each time point).

[0037] During the experiment, all rats were raised under unified conditions: temperature 22-24 ℃, humidity 45-55%, illumination conditions of 12 h light and 12 h dark cycle, rats could freely move and obtain food and water, and all experiments followed the 3R principle of experimental animal ethics.

[0038] Among them, before exposure treatment, the rats in each group were placed in the irradiation box for 30 min every day, and after 5 days of continuous adaptation, the exposure treatment (10 consecutive days) began. During the exposure process, the rats were placed in the special circular irradiation box, arranged at equal intervals, and the circular irradiation box was fixed on the radiation table. After the radiation was completed, the rats in each group were returned to the feeding cage. The treatment conditions of rats in each group were as follows: Sham exposure group: the power of the electromagnetic source and the noise source in this group was turned off, and the remaining conditions were the same as those in other treatment groups; Electromagnetic exposure group: rats in this group were exposed to electromagnetic exposure, and S-band and X-band microwaves were used to uniformly irradiate the rats. The power density was 10 mW / cm 2 , and the irradiation time for each frequency band was 10 min; Noise exposure group: rats in this group were exposed to noise, and the sound pressure level was 90 dB SPL (low-frequency noise) for 30 min; Electromagnetic and noise combined exposure group: rats in this group were first exposed to electromagnetic exposure, and then exposed to noise. The treatment conditions of the two were consistent with the corresponding treatment conditions of the electromagnetic exposure group and the noise exposure group.

[0039] 2. Behavior test On the 1st, 2nd, and 3rd days after the end of long-term exposure treatment, the effects of electromagnetic and noise exposure on the emotions and cognitive functions of rats in each group were evaluated through open field, new object, and Y maze experiments. The behavior test experimental process design is shown in Figure 1 .

[0040] (1) Open field experiment Experimental apparatus: composed of open field reaction box (purchased from Shenzhen Ruivode Life Science and Technology Co., Ltd.), data acquisition and analysis system (purchased from Grobel Biological Technology Co., Ltd.), wherein the open field reaction box has a size of 100 cm x 100 cm x 40 cm, is divided into 16 small squares of 25 cm x 25 cm, and a camera is installed directly above the box for tracking the activity trajectory of the rat; Pre-experiment adaptation: 60 min before the experiment, the rats were placed in the laboratory to be tested to adapt to the environment; Start the experiment: place the rat in the center of one side of the open field with its back to the experimenter, and record for 5 min using Any-maze behavior analysis software; after the test of the previous rat is completed, remove the feces, and wipe the inner wall and the surrounding area with 75% ethanol to remove the remaining odor; replace the rat and repeat the above steps.

[0041] On the first day after exposure, the total distance, average speed, total number of times entering the central area, time entering the central area, moving distance in the central area, and anxiety index of the rats were detected to evaluate the influence of electromagnetic radiation and noise exposure on the mood and voluntary movement of the rats.

[0042] (2) New object experiment device Learning period: two identical cylinders were placed in the experimental device with a distance of 10 cm between the objects and the outer edge, and the rats were placed in the device from a position equidistant from the objects with their backs to the objects (all rats were placed in the box in the same position), and the exploration time of the rats on the two objects within 5 min was recorded; Test period: the time interval between the learning period and the test period was 1 h; after the learning period ended, one of the cylinders in the box was replaced with a cone; then the rats were placed in the box against the wall (all rats were placed in the box in the same position), and the test time was 5 min, and the time of the rats exploring the two objects was recorded respectively; after the test of the previous rat was completed, the feces was removed, and the inner wall, the surrounding area, and the placed objects were wiped with 75% ethanol to remove the remaining odor; replace the rat and repeat the above steps.

[0043] On the first day after exposure, the rats' exploration time of new objects, exploration distance of new objects, recognition index (RI), discrimination index (DI), and other indicators were detected to evaluate the influence of electromagnetic radiation and noise exposure on the learning and memory ability of the rats.

[0044] (3) Y maze experiment Experimental equipment: consists of a Y-maze (purchased from Shenzhen Ruiwode Life Technology Co., Ltd.) and an image acquisition and analysis system. The Y-maze has three arms: the start arm, the novel arm, and the other arm. Each arm has an angle of 120° and a length, width, and height of 50 cm, 10 cm, and 20 cm, respectively. A camera is installed directly above the enclosure, and its field of view can cover the entire interior of the Y-maze. Adaptation phase: Close the new arm with a baffle, place the rat facing the center into the starting arm, and record its exploration trajectory for 5 minutes; after the previous rat has finished testing, remove the feces and wipe the inner wall, sides and objects placed with 75% ethanol to remove the remaining odor; replace the rat and repeat the above steps; Testing phase: After 1 hour of acclimatization, the baffle was removed and the rat was placed in the starting arm at the same location. Any-maze behavioral software was used to record the data for 5 minutes. After cleaning, the rat was replaced and the experiment continued. The time, distance, and number of times rats entered each maze arm were recorded. By analyzing the number of times rats entered novel arms, the movement time, the movement distance, and the movement speed, the effect of electromagnetic radiation on rats' short-term working memory was evaluated.

[0045] 3. Blood index testing Serum samples were collected immediately after exposure (day 0), day 7 (day 7), day 14 (day 14), and day 28 (day 28). The levels of creatine kinase (CK), lactate dehydrogenase (LDH), cardiac troponin I (cTn-I), angiotensin (Ang), testosterone (T), growth hormone (GH), S100 calcium binding protein β (S100-β), and ubiquitin carboxyl terminal esterase-L1 (UCHL1) were measured according to the kit instructions.

[0046] The source information of the reagent kit is shown in Table 1.

[0047] Table 1. Information on the source of the reagent kits

[0048] 4. Serum non-targeted metabolomics detection The serum sample on the 7th day after exposure (7 d) was taken out from the -80 °C refrigerator, thawed on ice until there was no ice in the sample (all subsequent operations were performed on ice); after the sample was thawed, it was vortexed for 10 s and mixed, 50 μL of the sample was transferred to the corresponding numbered centrifuge tube; then 300 μL of 20 % acetonitrile methanol internal standard extraction solution (purchased from Fisher, item number 955-4) was added, vortexed for 3 min, centrifuged at 12000 r / min for 10 min at 4 °C; after centrifugation, 200 μL of supernatant was transferred to another corresponding numbered centrifuge tube, and placed in a -20 °C refrigerator for 30 min; then centrifuged at 12000 r / min for 3 min at 4 °C, 180 μL of supernatant was transferred to the corresponding sample bottle inner tube for chromatographic analysis, wherein the chromatographic conditions were set as follows: Chromatographic column: Waters ACQUITY Premier HSS T3 Column 1.8 µm, 2.1 mm * 100 mm; Mobile phase A: 0.1 % formic acid / water; mobile phase B: 0.1 % formic acid / acetonitrile; Instrument column temperature: 40 °C; flow rate: 0.4 mL / min; sample size: 4 μL; The original data of mass spectrometric analysis was converted to mzML format by ProteoWizard, and then the XCMS program was used for peak extraction, alignment, and retention time correction; peaks with a missing rate > 50 % in each group of samples were filtered, and blank values were filled with KNN + 1 / 5 minimum value (blank values > 50 % were filled with 1 / 5 minimum value, and blank values < 50 % were filled with KNN); the SVR method was used to correct the peak area; the screened peaks were identified by searching the laboratory self-built database, integrating public libraries (HMDB, PubChem, Metlin, and KEGG), and predicting libraries (MetDNA); finally, substances with a comprehensive score of 0.5 or more and a QC sample CV value of less than 0.3 were extracted, and then positive and negative modes were combined (retaining the substance with the highest qualitative level and the smallest CV value), to obtain the all_sample_data.xlsx file.

[0049] Differential metabolite screening: First, the log2 logarithmic transformation was performed on the metabolite abundance values, and then the differential metabolites were screened by combining univariate and multivariate statistical analysis PLS-DA methods. Specifically, in the univariate analysis, for two-group comparison, the metabolites meeting the normal distribution were analyzed by t-test, and the metabolites not meeting the normal distribution were analyzed by wilcox-test; for multi-group comparison, the metabolites meeting the normal distribution were analyzed by ANOVA, and the metabolites not meeting the normal distribution were analyzed by Kruskal-Wallis-test; PLS-DA analysis would obtain the VIP (Variable Important for the Projection) value of each metabolite; therefore, the screening criteria for differential metabolites were P < 0.05 and VIP > 1; and K-means clustering analysis was used to identify metabolite groups with similar response trends under long-term electromagnetic and noise single / joint exposure.

[0050] 5. Data analysis The effects of electromagnetic radiation and noise single / joint exposure on the brain, heart and reproductive function of rats and their regularities were evaluated by two-way ANOVA, considering the interaction between the two; one-way ANOVA or Kruskal-Wallis test was used to evaluate the differences between multiple groups, and multiple correction methods were used for pairwise comparison; and SPSS 22.0, Graph Prism V.9.0 and R V4.0.5 software were used for statistical analysis of data and visualization of results.

[0051] 6. Experimental results (1) Changes in behavioral and physiological indicators The behavioral test results of rats in each group are shown in Figure 2 .

[0052] The results show that: Open field test: electromagnetic and noise exposure had no significant effect on the total distance and average speed of rats in the open field, and there was no interaction effect (F1, 48=0.05, η2p=0.067); while electromagnetic and noise exposure had no interaction effect on the time of rats entering the central area (F1, 48=0.05, η2p=0.024), but the main effect of noise exposure was significant, which showed that noise exposure significantly reduced the time of rats entering the central area (F1, 48=0.05, η2p (N)=0.127; F1, 48=0.05, η2p (E)=0.088), long-term noise exposure could cause anxiety-like behavior in rats, but electromagnetic radiation had no independent or synergistic effect (A~C in P P P P Figure 2

[0053] ​​​​​New object experiment: There is an interaction between the effects of electromagnetic radiation and noise on the recognition index (RI) of rats (P<0.05, η2p=0.52), which shows that electromagnetic exposure significantly reduces RI under noise-free conditions, but does not further aggravate cognitive impairment under noise exposure. Figure 2 (D in the middle).

[0054] Y-maze test: Electromagnetic and noise exposures did not interact with the behavioral performance of rats in the Y-maze. P >0.05), but the main effect of electromagnetic exposure was significant, that is, electromagnetic exposure reduced the percentage of time rats spent exploring novel arms [ P <0.05, η2p (E) =0.136; P >0.05, η2p (N) =0.005] and percentage of travel distance [ P <0.05, η2p(E) =0.115; P The values ​​>0.05 and η2p (N) =0.088 are significantly reduced, indicating that electromagnetic exposure may lead to impaired spatial memory.

[0055] (2) Blood index testing The results of blood parameter tests for each group of rats are shown below. Figure 3 .

[0056] The results show: Regarding myocardial injury-related indicators: on day 7 after long-term exposure ( P <0.05, η2p=0.666), 14d ( P <0.05, η2p=0.464) and 28 d ( P <0.05, η2p=0.419, indicating an interactive effect between long-term electromagnetic and noise exposure on cTn-I. Specifically, under noise-free conditions, electromagnetic exposure significantly increases cTn-I levels; while under electromagnetic exposure, noise significantly decreases cTn-I levels. Figure 3 (A) In the 7th day after exposure, the main effect of long-term electromagnetic exposure was significant, resulting in a significant increase in LDH. P <0.05, η2p (E)=0.439]; while on the 14th day after exposure ( P <0.05, η2p=0.515) and 28th day ( P <0.05, η2p=0.581, indicating an interactive effect between long-term electromagnetic and noise exposure on LDH. Specifically, electromagnetic radiation alone increases LDH, while this increasing trend is suppressed by combined noise exposure. Figure 3 (B) In the 7th day after exposure, the main effect of long-term electromagnetic exposure was significant, causing a significant increase in CK levels. P<0.05, η2p (E)=0.753]; while on the 14th day after exposure ( P <0.05, η2p=0.513) and 28th day ( P <0.05, η2p=0.791), the effects of long-term electromagnetic and noise exposure on CK also show a significant interaction, exhibiting a similar variation pattern to LDH ( Figure 3 Analysis of myocardial injury-related indicators (C) showed that long-term electromagnetic and noise exposure can induce myocardial injury, but their combined effect is not a simple additive but presents a complex nonlinear interaction. Myocardial injury indicators began to show significant changes on the 7th day after exposure, especially in the electromagnetic radiation group, suggesting that exposure causes persistent myocardial injury.

[0057] Regarding cardiovascular function-related indicators: On day 7 after long-term exposure, the main effect of electromagnetic radiation was significant, resulting in a significant increase in Ang-II levels. P <0.05, η2p (E)=0.681]; on the 14th day ( P <0.05, η2p=0.695) and 28 d ( P <0.05, η2p=0.541), electromagnetic and noise exposures showed a significant interaction. Specifically, under noise-free conditions, electromagnetic radiation significantly increased Ang-II levels; however, under electromagnetic exposure, noise decreased Ang-II levels. From day 7 post-exposure, Ang-II levels significantly increased, with electromagnetic radiation being the main influencing factor, possibly participating in the body's stress response through enhanced vasoconstriction and pro-inflammatory mechanisms. Noise may have an inhibitory effect on the increase of Ang-II levels to some extent, suggesting a possible complex mutual regulatory mechanism between the two exposure factors. Figure 3 (D in the middle).

[0058] Regarding endocrine hormone-related indicators: on day 7 after long-term exposure ( P <0.05, η2p =0.296), 14 d (P<0.05, η2p=0.506) and 28 d ( P <0.05, η2p=0.843), the effects of electromagnetic radiation and noise on T are interactive, and both show that under noise-free conditions, electromagnetic exposure reduces the synthesis of T, while at 14 and 28 days, under electromagnetic exposure conditions, noise exposure increases the synthesis of T. Figure 3 (E in the text); on day 7 after long-term exposure, the main effect of electromagnetic exposure was significant. P <0.05, η2p (E)=0.436], which significantly increased GH; on the 14th day ( P <0.05, η2p=0.375) and 28 d (P <0.05, η2p=0.425), electromagnetic and noise exposure interacted, which showed that electromagnetic radiation reduced GH synthesis under noise condition; while under electromagnetic radiation condition, noise exposure increased GH synthesis ( Figure 3 F in the results.

[0059] In terms of neuron injury related indicators: on the 7th day after long-term exposure, the main effect of electromagnetic radiation was significant[ P <0.05, η2p (E)=0.740], which significantly increased the level of S100-β; on the 14th day, electromagnetic radiation and noise interacted ( P <0.05, η2p=0.226), which showed that electromagnetic radiation significantly increased the level of S100-β under the condition of with or without noise; on the 28th day, electromagnetic radiation and noise also interacted ( P <0.05, η2p=0.455), which showed that electromagnetic radiation increased S100-β under the condition of without noise; while under the condition of electromagnetic radiation, noise decreased S100-β ( Figure 3 G in the results. P <0.05, η2p=0.431) on the 7th day ( P <0.05, η2p=0.276) on the 14th day ( P <0.05, η2p=0.423), electromagnetic radiation and noise both significantly interacted, which showed that electromagnetic radiation significantly increased the level of UCHL1 under the condition of without noise; in addition, on the 7th day and the 28th day, it was also observed that noise exposure significantly reduced the level of UCHL1 under electromagnetic condition; the effects of long-term electromagnetic and noise exposure on brain damage indicators showed time-dependent changes, in which electromagnetic radiation was the main influencing factor, which could independently cause brain damage indicators to significantly increase at other time points except immediately after exposure, suggesting its continuous disturbance to the nervous system; while under the condition of combined exposure, electromagnetic and noise did not cause the synergistic increase of S100-β and UCHL1 levels, which suggested that the two exposure factors might affect the nervous system through different or interfering signal pathways, leading to interference or offset of the damage effect, so as to not show additive or enhanced effect ( Figure 4 H in the results.

[0060] (3) Serum non-targeted metabolomics analysis The statistical results of serum non-targeted metabolite qualitative and quantitative analysis are shown in Figure 5 .

[0061] The results show that: in the serum samples of rats in each group 7 days after exposure, 1322 metabolites were identified by non-targeted metabolomics, and after classification and annotation, it was found that Lipids and lipid-like molecules were the most important metabolite category in serum, accounting for 35.93% of the total number of metabolites.

[0062] The intersection of differential metabolites at different time points of serum samples is shown in Figure 6 .

[0063] The results show that: based on univariate and multivariate analysis, the serum metabolome of different exposure groups at each time point was compared, and 172 inter-group differential metabolites were screened out at the end of long-term exposure; 82 differential metabolites were screened out at 7 days; 87 differential metabolites were screened out at 14 days; 108 differential metabolites were screened out at 28 days.

[0064] The KEGG enrichment pathway results of differential metabolites at different time points of serum samples are shown in Figure 6 .

[0065] The results show that: KEGG pathway enrichment analysis of differential metabolites at each time point after long-term electromagnetic and noise exposure found that the metabolic pathways at different time points were significantly different in number and type, showing dynamic evolution characteristics. Specifically, at the end of long-term exposure, 172 differential metabolites were significantly enriched in 5 pathways. Although the number of differential metabolites was the largest, the number of enriched pathways was relatively small, suggesting that the metabolic response at this stage may be more extensive and not yet focused on specific functional pathways, with a certain degree of non-specificity (A in Figure 6 ); At 7 days after exposure, the number of differential metabolite enriched pathways was the largest, a total of 9, although the number of differential metabolites was relatively small (82), but the metabolites were more concentrated in specific biological pathways, suggesting that there may be early disturbance of functional metabolic networks at this stage, and it may be related to damage effects (B in Figure 6 ); At 14 days after exposure, 87 differential metabolites were screened out, and only 1 pathway was enriched, which may reflect that the metabolic response at this stage is in a transition or regulation state (C in Figure 6 ); At 28 days after exposure, 108 differential metabolites were screened out, and were significantly enriched in 5 pathways, showing that the effects of electromagnetic and noise can still cause a certain degree of metabolic disorder at the later stage of exposure (D in Figure 7 ); This shows that the differential metabolites at 7 days after long-term exposure are more concentrated in multiple key biological pathways, combined with the results of changes in biochemical and physiological function indicators, suggesting that it may be a key early effect time window for metabolic disturbance.

[0066] Heatmap and trend of differential metabolites clustering analysis on day 7 after exposure Figure 8 .

[0067] The results show that: using K-means clustering algorithm to cluster the differential metabolites between groups on day 7 after exposure, it is found that C3 (including 6 metabolites) and C4 (including 12 metabolites) two clusters of metabolites in the EN group showed a significant upward trend, while C9 (including 7 metabolites) metabolites in the EN group showed a significant downward trend.

[0068] The results of key metabolite and phenotype correlation analysis on day 7 after exposure (EN group) are shown in Figure 9 .

[0069] The results show that: the 25 metabolites significantly up-regulated or down-regulated in the EN group have significant correlation with multiple phenotype indicators, among which 1-Stearoyl-2-linoleoyl-sn-glycerol, LPA(18:2(9Z,12Z) / 0:0, PE-NMe(16:0 / 16:0), 14,15-Leukotriene C4(ExC4), Canrenone, 2-Oxovaleric acid, 2-Nitrofluorene are significantly negatively correlated with RI, and also significantly positively correlated with myocardial / brain injury indicators and cardiovascular function indicators, etc. This result further supports the potential indicative value of these metabolites in the process of nervous system injury, suggesting that they may serve as early biomarkers for identifying multiple system function (mainly neural and cardiovascular function) damage after exposure.

[0070] The results of focusing on the KEGG pathway in which the metabolites are involved are shown in Figure 9 .

[0071] The results show that: 1-Stearoyl-2-linoleoyl-sn-glycerol, LPA(18:2(9Z,12Z) / 0:0 and PE-NMe(16:0 / 16:0) are significantly correlated with RI, and also significantly correlated with myocardial and brain injury indicators, and are closely related to phosphoinositide metabolism, glycerolipid metabolism, glycerophospholipid metabolism, phosphatidylinositol signaling system, phospholipase D signaling pathway, gap junction, actin cytoskeleton regulation, and long-term depression (A in Figure 9 , further supporting the possibility that they are involved in the process of cognitive function impairment caused by complex exposure; visual analysis of the dynamic trends of these three metabolites on day 0, 7, 14 and 28 after long-term exposure found that the differences between the candidate metabolites and the control group were most significant on day 7 ( ​The results showed that the changes of these metabolites were mainly concentrated in the early stage after exposure ended, which was consistent with the abnormal changes of brain injury and behavioral phenotype on the 7th day and further supported the potential of these metabolites as biomarkers of early injury.

[0072] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "an example", "a specific example", or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. Furthermore, different embodiments or examples described in the present specification and the features of different embodiments or examples can be combined and combined by those skilled in the art without contradiction.

[0073] Although the embodiments of the present application have been shown and described above, it is understood that the above-described embodiments are exemplary and are not to be construed as limiting the present application, and those skilled in the art can make changes, modifications, replacements and variations to the above-described embodiments within the scope of the present application.

Claims

1. A biomarker for risk assessment of an individual's exposure to electromagnetic and / or noise, characterized in that, The markers include one or more of 1-stearoyl-2-linoleoyl-sn-glycerol, 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid, and N,N-dimethylphosphatidylethanolamine (16:0 / 16:0).

2. The use of reagents for detecting 1-stearoyl-2-linoleoyl-sn-glycerol, reagents for detecting 1-(9Z,12Z-octadecadienoyl)-lysophosphatidic acid, or reagents for detecting N,N-dimethylphosphatidylethanolamine (16:0 / 16:0) in the preparation of a kit, characterized in that, The kit is used for risk assessment of individuals exposed to electromagnetic and / or noise.

3. A kit for risk assessment of an individual's exposure to electromagnetic and / or noise, characterized in that, The kit contains reagents for detecting the marker of claim 1.

4. A method for assessing an individual's electromagnetic and / or noise exposure risk, characterized in that, The method includes: Obtain a sample to be tested and detect the abundance level of the biomarker of claim 1 in the sample to be tested, and assess the risk level of the individual's exposure to electromagnetic and / or noise environment based on the abundance level; The method is used for non-diagnostic purposes.

5. The method according to claim 4, characterized in that, The sample to be tested is a blood sample.

6. A system or apparatus, characterized in that, The system or apparatus includes: A data acquisition module is used to acquire abundance data of the biomarker of claim 1 in samples from individuals; The risk assessment module is used to assess the degree of risk of the individual's exposure to electromagnetic and / or noise based on the abundance data.

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