Mouse hangover degree accurate detection mathematical analysis model construction method and application

Abstract

The invention provides a mouse hangover degree accurate detection mathematical analysis model construction method and application. Specifically, the invention provides a method for constructing a hangover degree accurate detection mathematical analysis model, and the method comprises the following steps: carrying out wine gavage and ethanol gavage on a mouse, carrying out an open field experiment and a rotating bar experiment, and detecting the concentration of ethanol in the blood of the mouse, so as to obtain each index numerical value, thereby obtaining the hangover degree of the mouse. The method disclosed by the invention is simple and convenient to operate and high in credibility and sensitivity, and has important significance in actual production and application of the wine industry.

Description

Technical Field

[0001] The present invention relates to the field of medical diagnosis, and in particular to a method for constructing a mathematical analysis model for accurately detecting the degree of hangover in mice and its application. Background Art

[0002] According to the latest World Health Organization report on global drinking and health, as the world economy develops, the total per capita alcohol consumption of adults worldwide reached 6.4 liters of pure alcohol by 2016, while in my country this level reached 7.2 liters of pure alcohol per person per year. And according to the model prediction, the per capita alcohol consumption level worldwide will continue to grow, especially in developing countries like my country. Under such a high level of alcohol consumption, the health and social security problems caused by drinking are particularly prominent around the world, which has caused a huge burden on global public health.

[0003] Ethanol is a highly polar, fat-soluble substance that can quickly enter the blood circulation in a very short time. It is mainly absorbed in trace amounts by the oral mucosa, a small part by the stomach (about 10%-20%), and most of it by the small intestine. More than 90% of the ethanol in the human body is metabolized in the liver. Under the action of liver alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH), ethanol is decomposed into acetic acid. When ethanol enters the human body, as the blood concentration of ethanol gradually increases, the body begins to relax slowly and enters an excited state; and when its level further increases, with a large amount of ethanol crossing the blood-brain barrier and entering the brain, the body presents symptoms of alcohol intoxication such as ataxia and coma. When ethanol is completely cleared from the body (i.e., the drunken stage ends), the body is still in a more persistent physiological and psychological negative state, such as fatigue, weakness, nausea, vomiting, thirst and other symptoms. This symptom is called a hangover, which will last for several hours or even 24 hours. When in a hangover state, the body's reaction, cognition and motor ability are significantly reduced. Although people have been drinking for more than 12 hours, their physical condition has not yet returned to normal, which brings great burdens to human health and social security.

[0004] At present, with the rapid development of the national economy, people are paying more and more attention to health, especially food health. Alcohol is a food. After satisfying the "color" and "aroma" before drinking, and the "taste" and "style" during drinking, people's pursuit of alcohol quality has gradually turned to the comfort of the body after drinking. The comfort of good alcohol after drinking usually means no dry mouth, no intoxication, and no hangover after drinking, among which "no hangover" occupies an important position.

[0005] Mice are the most commonly used animal model in animal experiments. They are easy to raise, have a high reproduction rate, are genetically pure, have small differences between individuals, and are easy to observe the results of parallel experiments. Their metabolic types and physiological and pathological characteristics are similar to those of humans. Most importantly, the ethanol metabolism pattern of mice is very similar to that of humans, making them an ideal animal model for detecting hangover levels.

[0006] Therefore, it is of great significance to establish an effective method and its application to accurately and efficiently evaluate the degree of alcohol hangover using mice as model animals. Summary of the invention

[0007] The invention provides an effective method for accurately and efficiently evaluating the hangover degree of alcoholic beverages and its application.

[0008] In a first aspect of the present invention, a method for constructing a mathematical analysis model for accurately detecting hangover degree is provided, comprising the steps of:

[0009] (S1) Provide two groups of mice with the same origin and baseline conditions and raise them under the same environmental conditions;

[0010] (S2) administering alcohol orally to one group of mice, and administering ethanol orally to another group of mice;

[0011] (S3) performing an open field test on the mice in the alcohol gavage group and the mice in the ethanol gavage group, respectively, and recording the probability P1 that the distance moved by the mice in the alcohol gavage group in the open field is smaller than the distance moved by the mice in the ethanol gavage group in the open field;

[0012] (S4) performing a rotating rod test on the mice in the alcohol gavage group and the mice in the ethanol gavage group, respectively, and recording the probability P2 that the rotating rod time of the mice in the alcohol gavage group is shorter than that of the mice in the ethanol gavage group;

[0013] (S5) respectively testing the blood ethanol concentration of the mice in the wine gavage group and the ethanol gavage group, and recording the probability P3 that the blood ethanol concentration of the mice in the wine gavage group is greater than the blood ethanol concentration of the mice in the ethanol gavage group; and

[0014] (S9) Based on each recorded probability, different weights W are assigned to each recorded probability according to the correlation between each detection index and the degree of hangover i ; Establish a mathematical analysis model for accurate detection of hangover degree, so as to obtain the hangover index (Hangover Index) P:

[0015]

[0016] Among them, step (S3), step (S4), and step (S5) can be performed in any order.

[0017] In another preferred embodiment, the method may further optionally perform the following steps:

[0018] (S6) respectively detecting the blood acetaldehyde concentration of the mice in the alcohol gavage group and the mice in the ethanol gavage group, and recording the probability P4 that the blood acetaldehyde concentration of the mice in the alcohol gavage group is lower than the blood acetaldehyde concentration of the mice in the ethanol gavage group;

[0019] (S7) respectively detecting the concentration of acetaldehyde dehydrogenase (ALDH) in the liver of the mice in the alcohol gavage group and the mice in the ethanol gavage group, and recording the probability P5 that the concentration of ALDH in the liver of the mice in the alcohol gavage group is lower than that of the mice in the ethanol gavage group;

[0020] (S8) Detecting cerebellar gamma-aminobutyric acid (GABA) in the mice in the alcohol gavage group and the mice in the ethanol gavage group, respectively, and recording the probability P6 that the cerebellar GABA level in the mice in the alcohol gavage group is greater than the cerebellar GABA level in the mice in the ethanol gavage group.

[0021] In another preferred embodiment, the experiment and detection of the method are carried out after mice are gavaged with ethanol or alcohol for a period of time.

[0022] In another preferred embodiment, the experiment and detection of the method are carried out 6-8 hours after mice are gavaged with ethanol or alcohol.

[0023] In another preferred embodiment, the experiment and detection of the method are carried out 6 hours after the mice are gavaged with ethanol or alcohol.

[0024] In another preferred embodiment, the experiment and detection of the method are carried out 7 hours after the mice are gavaged with ethanol or alcohol.

[0025] In another preferred embodiment, in the intragastric administration of wine and ethanol, the ethanol content of the wine and ethanol is the same.

[0026] In a second aspect of the present invention, there is provided an evaluation system for detecting the degree of hangover, the system comprising:

[0027] (M1) Input module: The input module is configured to input open field test data, rotating rod test data and blood alcohol concentration data of the subject to be tested;

[0028] (M2) Evaluation module: The evaluation module is configured to evaluate the open field test data, the rotating rod test data, and the blood ethanol concentration data based on the mathematical analysis model for accurate detection of the degree of hangover, so as to obtain an evaluation result of the degree of hangover of the subject to be tested; wherein the mathematical analysis model for accurate detection of the degree of hangover is a mathematical analysis model for accurate detection of the degree of hangover constructed by the construction method described in the first aspect of the present invention; the model formula is:

[0029] and

[0030] (M3) Output module: The output module is configured to output the evaluation result.

[0031] In another preferred example, the system further includes: (M4) a control module: the control module is configured to control the operation of each module.

[0032] In another preferred embodiment, the greater the evaluation result value of the evaluation, the higher the degree of hangover; conversely, the smaller the evaluation result value of the evaluation, the lower the degree of hangover.

[0033] In another preferred embodiment, the test subjects include: ethanol-administered mice and alcohol-administered mice.

[0034] In another preferred embodiment, the alcoholic beverage includes types selected from the following group: white wine, yellow wine, fruit wine, beer, rice wine, medicinal wine, or a combination thereof.

[0035] In another preferred example, the data input by the input module further includes data selected from the following group: blood acetaldehyde concentration, cerebellar GABA level, liver ALDH concentration, or a combination thereof.

[0036] In a third aspect of the present invention, there is provided an evaluation method, comprising the steps of:

[0037] (a) providing the open field test data, rotarod test data and blood ethanol concentration data of the test subject;

[0038] (b) inputting the open field test data, the rotating rod test data and the blood ethanol concentration data into the mathematical analysis model for accurate detection of hangover degree constructed by the construction method described in the first aspect of the present invention, wherein the model formula is:

[0039]

[0040] (c) using the mathematical analysis model for accurate detection of hangover degree to evaluate the open field test data, the rotarod test data and the blood ethanol concentration data, thereby obtaining the hangover degree of the subject to be tested; and

[0041] (d) Output the evaluation results.

[0042] In another preferred embodiment, the greater the evaluation result value of the evaluation is, the higher the degree of hangover is; conversely, the smaller the evaluation result value is, the lower the degree of hangover is.

[0043] In another preferred embodiment, the test subjects include: ethanol-administered mice and alcohol-administered mice.

[0044] In another preferred embodiment, the evaluation method is non-diagnostic and non-therapeutic.

[0045] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 The evaluation indicators of the accurate detection method for the degree of hangover in mice are shown: (A) 6 hours after the mice were gavaged with ethanol, behavioral tests were performed: open field and rotarod tests, cerebellar tissue GABA measurement, blood ethanol and acetaldehyde concentration measurement, liver ADH and ALDH activity measurement.

[0047] Figure 2 The figure shows that 6 h after intragastric administration of ethanol was the time point for detecting the degree of hangover in mice: the moving distance in the open field (A, B), the time on the rotarod (C), the blood ethanol (D) and acetaldehyde (E) concentrations, the cerebellar GABA content (F), and the liver ADH (G) and ALDH (H) activities were measured at 1, 2, 4, 6, 8, 10, and 24 h after intragastric administration of 2.5 g / kg ethanol in mice; the normal saline intragastric administration group was taken as 0 h.

[0048] Figure 3 The mathematical analysis model for accurately detecting the degree of hangover in mice is shown: (A) Taking the ethanol group as the control, the probability that each indicator in the test group is greater than or less than the corresponding indicator is calculated. According to the weight, the weighted sum of each branch is taken to obtain the final hangover index (Hangover Index), and it is stipulated that the larger the index, the higher the degree of hangover.

[0049] Figure 4 The model was used to detect the degree of hangover of alcohol samples: mice were given simple alcohol, saline and a certain liquor sample (same dose) by gavage for 1 hour, and the distance moved by the mice in the open field was measured ( Figure 4 AB), the time of rotating rod on the rod ( Figure 4 C), blood alcohol ( Figure 4 D) concentration and acetaldehyde ( Figure 4 E) concentration, cerebellar GABA level ( Figure 4 F) and liver ALDH activity ( Figure 4G); Based on the above indicators, the mathematical analysis model was used to calculate the hangover index (Hangover Index) of ethanol, saline and a certain liquor sample. Figure 4 H). DETAILED DESCRIPTION

[0050] After extensive and in-depth research, the inventors unexpectedly discovered a method for evaluating the degree of hangover. Specifically, the inventors established a mathematical analysis model for accurately predicting the degree of hangover through three evaluation indicators (behavior, neurotransmitters, and biochemistry) after mice were gavaged with white wine. The behavior included test results such as the Open Field Test (OFT) and the Rotarod Test, neurotransmitters included cerebellar gamma-aminobutyric acid (GABA) levels, and biochemical indicators included blood ethanol, blood acetaldehyde concentration, and liver acetaldehyde dehydrogenase (ALDH) activity. The analysis model of the present invention can better evaluate the degree of hangover. On this basis, the present invention was completed.

[0051] Method for establishing mathematical analysis model for accurate detection of hangover degree

[0052] The present invention provides a method for constructing a mathematical analysis model for accurately detecting the degree of hangover, comprising the following steps:

[0053] (S1) Provide two groups of mice with the same origin and baseline conditions and raise them under the same environmental conditions;

[0054] (S2) administering alcohol orally to one group of mice, and administering ethanol orally to another group of mice;

[0055] (S3) performing an open field test on the mice in the alcohol gavage group and the mice in the ethanol gavage group, respectively, and recording the probability P1 that the distance moved by the mice in the alcohol gavage group in the open field is smaller than the distance moved by the mice in the ethanol gavage group in the open field;

[0056] (S4) performing a rotating rod test on the mice in the alcohol gavage group and the mice in the ethanol gavage group, respectively, and recording the probability P2 that the rotating rod time of the mice in the alcohol gavage group is shorter than that of the mice in the ethanol gavage group;

[0057] (S5) respectively testing the blood ethanol concentration of the mice in the wine gavage group and the ethanol gavage group, and recording the probability P3 that the blood ethanol concentration of the mice in the wine gavage group is greater than the blood ethanol concentration of the mice in the ethanol gavage group; and

[0058] (S9) Based on each recorded probability, different weights W are assigned to each recorded probability according to the correlation between each detection index and the degree of hangover i; Establish a mathematical analysis model for accurate detection of hangover degree, so as to obtain the hangover index (Hangover Index) P:

[0059]

[0060] Among them, step (S3), step (S4), and step (S5) can be performed in any order.

[0061] In another preferred embodiment, the method may further optionally perform the following steps:

[0062] (S6) respectively detecting the blood acetaldehyde concentration of the mice in the alcohol gavage group and the mice in the ethanol gavage group, and recording the probability P4 that the blood acetaldehyde concentration of the mice in the alcohol gavage group is lower than the blood acetaldehyde concentration of the mice in the ethanol gavage group;

[0063] (S7) respectively detecting the concentration of acetaldehyde dehydrogenase (ALDH) in the liver of the mice in the alcohol gavage group and the mice in the ethanol gavage group, and recording the probability P5 that the concentration of ALDH in the liver of the mice in the alcohol gavage group is lower than that of the mice in the ethanol gavage group;

[0064] (S8) Detecting cerebellar gamma-aminobutyric acid (GABA) in the mice in the alcohol gavage group and the mice in the ethanol gavage group, respectively, and recording the probability P6 that the cerebellar GABA level in the mice in the alcohol gavage group is greater than the cerebellar GABA level in the mice in the ethanol gavage group.

[0065] In another preferred embodiment, the experiment and detection of the method are carried out after mice are gavaged with ethanol or alcohol for a period of time.

[0066] In another preferred embodiment, in the intragastric administration of wine and ethanol, the ethanol content of the wine and ethanol is the same.

[0067] Hangover assessment system (equipment)

[0068] The present invention provides an evaluation system (device) for detecting the degree of hangover, the system comprising:

[0069] (M1) Input module: The input module is configured to input open field test data, rotating rod test data and blood alcohol concentration data of the subject to be tested;

[0070] (M2) Evaluation module: The evaluation module is configured to evaluate the open field test data, the rotating rod test data, and the blood ethanol concentration data based on the mathematical analysis model for accurate detection of the degree of hangover, so as to obtain an evaluation result of the degree of hangover of the subject to be tested; wherein the mathematical analysis model for accurate detection of the degree of hangover is a mathematical analysis model for accurate detection of the degree of hangover constructed by the construction method described in the first aspect of the present invention; the model formula is:

[0071] and

[0072] (M3) Output module: The output module is configured to output the evaluation result.

[0073] In another preferred example, the system further includes: (M4) a control module: the control module is configured to control the operation of each module.

[0074] In another preferred embodiment, the greater the evaluation result value of the evaluation, the higher the degree of hangover; conversely, the smaller the evaluation result value of the evaluation, the lower the degree of hangover.

[0075] In another preferred embodiment, the test subjects include: ethanol-administered mice and alcohol-administered mice.

[0076] In another preferred example, the data input by the input module further includes data selected from the following group: blood acetaldehyde concentration, cerebellar GABA level, liver ALDH concentration, or a combination thereof.

[0077] The main advantages of the present invention include:

[0078] 1. The present invention is the first to establish an accurate detection model for the degree of hangover in mice based on the laws of ethanol metabolism, which is highly similar to the characteristics of symptoms after drinking alcohol in the human body, including the detection time point and evaluation and analysis methods, etc., which will greatly fill the gap in this field and incorporate the degree of hangover into the comfort evaluation system after drinking alcohol.

[0079] 2. The mathematical analysis model for hangover detection of the present invention can comprehensively, truly and objectively reflect the overall hangover state of mice, and can well distinguish the subtle differences between samples in each group, and the model has high credibility and sensitivity.

[0080] 3. The mathematical analysis model for accurately detecting the degree of hangover in mice of the present invention is simple and convenient to operate and is less susceptible to interference.

[0081] 4. The present invention has a wide range of application scenarios. In addition to being used for the evaluation of finished wine samples, it can also be used to evaluate wine samples in the production process, such as base wine, blended wine, etc.; it can also be used to study the influence of flavor substances on the degree of hangover, so as to improve the quality of wine; it can also evaluate the efficacy of alcohol-detoxifying drugs, detect the degree of hangover of mice after drug administration, etc.

[0082] 5. The mathematical analysis model for accurately detecting the degree of hangover in mice of the present invention has low actual use cost and meets the needs of large-scale use by enterprises.

[0083] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are weight percentages and weight parts.

[0084] Materials and methods

[0085] Experimental animals

[0086] Eight-week-old male C57Bl / 6 mice were purchased from Shanghai Slack Company. They were adapted to the standard SPF environment for one week before the formal experiment and fed with standard food during this period.

[0087] Example 1 Mouse hangover experiment

[0088] 1.1 Alcohol gavage in mice

[0089] The ethanol gavage dose for mice was 2.5 g / kg body weight, the ethanol concentration was 31.5% (v / v), and the gavage volume was 10 times the body weight of the mice. A disposable sterile syringe combined with a sterilized No. 8 gavage needle was used for gavage to ensure the gavage effect, no secondary damage to the mice, and no choking of ethanol solution.

[0090] After intragastric administration, the mice were placed in cages to rest and wait for testing. The ambient temperature was controlled at around 25°C to ensure that the mice did not suffer from hypothermia.

[0091] The alcohol solution for intragastric administration of mice was prepared with chromatographic grade anhydrous ethanol (Aladdin, E111993-500ml). The alcohol concentration of the intragastric alcohol sample was measured with an alcohol meter and then diluted to 31.5% (v / v).

[0092] After the experiment, the mice were killed by an overdose of isoflurane, blood was drawn from the apex of the heart, and liver and cerebellum tissues were collected, weighed, and quickly frozen in liquid nitrogen for testing.

[0093] 1.2 Behavioral experiments

[0094] 1.2.1 Open field test for mice

[0095] The open field video analysis system (SA215, Jiangsu Seons) was used to perform open field tests on mice and analyze video data. Before the experiment, the mice were placed in the open field equipment room to adapt to the environment for half an hour. The total moving distance of the mice in the open field (50 cm × 50 cm) for 20 minutes was recorded and analyzed by video. Four mice could be tested and analyzed at the same time.

[0096] 1.2.2 Mouse Rotarod Test

[0097] The mouse rotarod fatigue test was performed using a mouse rotarod fatigue instrument (Jiangsu Seons, SA102). The rotarod setting parameters were an initial speed of 4 r / min, accelerated to 40 r / min within 5 min, and an acceleration of 7.2 r / min. 2 The mice need to be trained on the rotarod continuously for three days before the formal test, three times a day, with an interval of more than 30 minutes between each training. The mice that have an average time on the rod of more than 200 seconds on the last day can be used for the formal test experiment. Before the experiment, the mice need to adapt to the environment for more than 30 minutes.

[0098] Start the rotating rod, place the mice on the rotating rod one by one at the initial speed, and then start to accelerate. The instrument automatically records the time when the mice fall.

[0099] 1.3 Determination of GABA levels in mouse cerebellum

[0100] 1.3.1 GABA extraction from cerebellum

[0101] About 50 mg of mouse cerebellum tissue was added with 300 μL ultrapure water and then ground into a homogenate. Then 700 μL methanol and acetonitrile (chromatographic grade) solution (1:1) was added, and the mixture was shaken for 30 seconds and then ultrasonicated in an ice bath. Then, it was placed in a -20°C refrigerator for 1 hour, centrifuged at 12000 rpm for 15 minutes at 4°C, and the supernatant was transferred to a new 1.5 mL centrifuge tube. All liquid in the supernatant was evaporated using a vacuum freeze dryer, and 300 μL acetonitrile aqueous solution (1:9) was added to dissolve it, and then it was shaken for 30 seconds and then ultrasonicated in an ice bath. Then, it was centrifuged again at 12000 rpm for 15 minutes at 4°C, the supernatant was collected, and it was diluted 50 times with ultrapure water and then tested on the machine.

[0102] 1.3.2 GABA assay

[0103] GABA was determined by liquid chromatography-mass spectrometry, and the instrument information is as follows: liquid chromatography system: Agilent 1200 LC system (binary pump, automatic sampler with temperature control device, column oven, online degasser), mass spectrometry system: SCIEX 4000 QTRA PLC / MS / MS systems.

[0104] Chromatographic column: Agilent Zorbox Eclipse Plus C18 4.6*150mm, 5um;

[0105] Mobile phase A: water + 0.1% formic acid + 0.01% heptafluorobutyric acid;

[0106] Mobile phase B: methanol + 0.1% formic acid + 0.01% heptafluorobutyric acid, flow rate: 0.8 ml / min;

[0107] Column temperature: 50°C. Gradient elution: 0-3min: 2% B, 3-5min: 20% B, 5-5.5min: 90% B, 5.5-7.5min: 90% B, 7.5-8min: 2% B, 8-10min: 2% B;

[0108] Mass spectrometry parameters: ion source: point spray ion source (EIS), positive ion multiple reflection detection (MRM) mode scanning, capillary voltage (IS) was 5500 V, ion source temperature (TEM) was 600 °C, curtain gas (CUR) was 15 psi, collision gas (CAD) was 8 psi, nebulizer gas (GS1) was 60 psi, auxiliary gas (GS2) was 50 psi, declustering voltage (DP) was 43 V, inlet voltage was 10 V, collision cell exit voltage (CXP) was 7 V, and collision voltage (CE) was 15 V;

[0109] Quantitative ion pair: m / z 104.1→87.1, retention time 3.5 min.

[0110] 1.4 Determination of biochemical indicators in mice

[0111] 1.4.1 Determination of blood ethanol and acetaldehyde concentrations

[0112] Headspace gas chromatography (Agilent 7890BGC with FID, Agilent 7697A headspace sampler) was used to determine the ethanol and acetaldehyde content in the blood. 0.4 mL of blood was taken, 0.1 mL of tert-butyl alcohol solution was added as an internal standard, and 0.5 mL of ultrapure water was added to a 20 mL headspace bottle, which was sealed with a crimp clamp.

[0113] The chromatographic column was Agilent J&WDB-BAC1UI, 30m×0.32mm, 1.8m, column temperature 40℃, lasting 5min. The carrier gas was nitrogen, constant flow, 6mL / min. The injection port temperature was 210℃, split mode, split ratio 1:1. FID conditions: temperature was 250℃, hydrogen flow rate was 30mL / min, air flow rate was 350mL / min, and tail blowing was 35mL / min. HS conditions: column oven 70℃ (equilibration 15min), quantitative loop 80℃, transfer line 90℃. The retention times of acetaldehyde, acetaldehyde, and tert-butyl alcohol were 1.2, 1.5, and 2.0min, respectively.

[0114] 1.4.2 Determination of liver ADH and ALDH activity

[0115] The enzyme activities were determined using ADH (Nanjing Jiancheng, A083-2-1) and ALDH (Solarbio, A083-2-1) activity assay kits, respectively, and the relevant steps were based on their instructions.

[0116] 1.5 Experimental Results

[0117] 1.5.1 6 h after ethanol gavage as the time point for detecting the degree of hangover in mice

[0118] When the body is in a hangover state, its cognitive, motor and coordination functions are significantly reduced.

[0119] If the degree of hangover is mild, the effect of ethanol on mice cannot be fully reflected; if the mice are completely hangover and unconscious, the differences between the alcohol samples and the effect of ethanol on the motor ability of mice may not be reflected. Therefore, finding a suitable degree of hangover for mice is crucial for the evaluation of hangover degree.

[0120] Based on this, mice were given 2.5 g / kg ethanol by gavage, and the changes of various indicators at each time point were monitored for a continuous period of 24 hours. The results are as follows Figure 2 shown.

[0121] In behavioral tests, the movement and coordination abilities of mice showed a two-phase distribution with the time after ethanol gavage: first decreased and then recovered (0-2h), then decreased again and then recovered (2-24h) ( Figure 2 A, B and C). This is closely related to the blood ethanol concentration of mice. The blood ethanol concentration of mice was close to zero (<100 mg / L) 6 hours after oral gavage ( Figure 2 D) When the ethanol in the mouse is completely metabolized, the mouse enters a hangover state. At this time, the intermediate metabolite of ethanol, acetaldehyde, has not been completely decomposed ( Figure 2 E), acetaldehyde takes over the role of ethanol and continues to have a negative impact on the human body. Hangover occurs at 6 hours, and the body movement and coordination ability of mice are the lowest in this stage. The GABA level in the mouse cerebellum is similar to the change of blood ethanol, showing an increase first and then a decrease as the ethanol concentration decreases ( Figure 2 F). The ADH activity in mouse liver did not change significantly over time ( Figure 2 G), it has completed the ethanol decomposition metabolism at the hangover stage, so it is not suitable for the hangover assessment. However, when the hangover occurs (6h), its activity increases ( Figure 2 H), continuously catabolizes acetaldehyde.

[0122] Based on the changes in the above indicators, it was found that 6 hours after ethanol gavage, the ethanol concentration in the body of mice was close to zero, and their movement and coordination abilities were low. At the same time, there was still a large amount of acetaldehyde remaining in the blood to be decomposed, and the liver ALDH activity was vigorous. Therefore, this time point is more suitable as a time point for detecting the degree of hangover.

[0123] Example 2 Mathematical analysis model for detecting the degree of hangover in mice

[0124] 2.1 Construction of mathematical analysis model for hangover detection

[0125] In order to better reflect the degree of hangover, based on the above evaluation indicators, this patent establishes a hangover index (Hangover Index) mathematical model that can integrate the characteristics of the above indicators.

[0126] Specifically, the pure ethanol group was used as the control group, and each wine sample group was used as the test group. The probability (P1-P6) that the control group of each indicator in Example 1 was greater than or less than the test group was calculated, and then certain weights (W1-W6) were assigned to the corresponding probabilities of these indicators. Finally, the probability of each indicator was weighted and summed to obtain the final probability, which was defined as the Hangover Index. It was stipulated that the larger the probability, the greater the degree of hangover.

[0127] The specific analysis model is as follows Figure 3 shown.

[0128] Taking the pure ethanol group as the control, the various indicators of the test group were compared with those of the control group, the probability of each indicator was calculated, and the different probabilities were assigned certain weights and then weighted summed.

[0129] Specifically, the total moving distance of mice in the open field was defined as O, the control group and the test group were O0 and O1, where O 01 -o 0n and 11 -o 1n Represents the total moving distance of different mice in the control group and the test group in the open field. Similarly, the time on the rod in the mouse rotarod test, blood ethanol concentration, blood acetaldehyde concentration, cerebellar GABA level, and liver ALDH activity were defined as R, B, C, G, and A, respectively.

[0130] Next, compare the above two groups of indicators and define p1 = P(O0>O1), p2 = P(R0>R1), p3 = P(B0<B1),p4=P(C0> C1), p5=P(G0<G1),p6=P(A0> A1), and define the weights of p1–p6 as w1=w2=0.3, w3=w4=0.15, w5=w6=0.05, respectively. The specific ratio of w1-w6 is 6:6:3:3:1:1, and the sum of w1-w6 is 1.

[0131] Finally, p1–p6 are weighted and summed according to the weights w1-w6, that is p is defined as the Hangover Index, with a value between 0 and 1. The p of the control group is approximately 0.5. The smaller the p of the test group, the lower the degree of hangover.

[0132] The above calculation process can be quickly implemented using R language.

[0133] 2.2 Detection of hangover degree in mice and application of mathematical analysis model

[0134] 2.2.1 Experimental methods

[0135] The patented model was applied using physiological saline (positive control) and liquor samples with the same dose of ethanol to verify the sensitivity and reliability of the model.

[0136] With the pure ethanol group as the control, the mice were gavaged with the three groups of samples with the same ethanol dose, and the relevant indicators were measured 6 hours after gavage.

[0137] 2.2.2 Experimental results

[0138] The results are as follows Figure 4 In terms of the motor ability of mice, their motor ability was significantly reduced after being gavaged with alcohol; however, the motor phenotype of the alcohol-like mice gavaged with the same dose of ethanol was better than that of the ethanol control group, and their moving distance in the open field and the time they stayed on the rotating rod were significantly higher than those of the ethanol control group ( Figure 4 A, B and C). Similarly, the blood ethanol concentration of mice in the liquor sample group was lower than that in the ethanol control group ( Figure 4 D), indicating that it may have a faster ethanol metabolism rate. However, there was no significant change in blood acetaldehyde concentration, cerebellar GABA level and liver ALDH activity at this time ( Figure 4 E).

[0139] After determining the model detection index of this patent, the mathematical analysis model is used to quickly calculate the probability value (p1-p6) corresponding to each group of indicators and the final hangover index: Hangover Index ( Figure 4 G), as shown in Table 1.

[0140] Table 1

[0141] EtOH Normal saline Liquor <![CDATA[p1]]> 0.44 0.00 0.00 <![CDATA[p2]]> 0.43 0.00 0.14 <![CDATA[p3]]> 0.42 0.00 0.18 <![CDATA[p4]]> 0.44 0.00 0.55 <![CDATA[p5]]> 0.44 0.41 0.47 <![CDATA[P6]]> 0.44 0.47 0.34 Hangover Index 0.43 0.04 0.19

[0142] The Hangover Index of the saline group was close to 0, indicating that the hangover degree detection and mathematical analysis model based on this patent can well distinguish whether the mice have drunk alcohol, reflecting that the model of this patent has high reliability; and the Hangover Index of a certain liquor sample was 0.19, which was lower than 0.43 of the ethanol group, indicating that the mathematical analysis model of this patent can better integrate and quantify the data characteristics of each detection index, reflecting the hangover degree of the liquor sample.

[0143] 2.2.3 Comparative experiments and results

[0144] Given that the weights of the given indicators are equal and their sum is 1, if only the open field test P1, only the rotarod test P2, only the two behavioral tests p1 and p2, and only P1, P2, P3, and P4 are calculated, the Hangover Index of a certain liquor is 0, 0.14, 0.07, and 0.22 respectively.

[0145] In this case, the model cannot fully reflect the overall hangover state of mice, and cannot distinguish the subtle differences between the samples in each group. The sensitivity of the overall model will be significantly reduced, and large errors will occur in actual application.

[0146] In summary, the accurate detection method and mathematical analysis model of hangover degree described in this patent have high reliability and sensitivity, and can be used for accurate detection of hangover degree in the actual alcohol industry.

[0147] discuss

[0148] There are currently no behavioral studies on evaluating the extent of hangover in mice.

[0149] Existing patents usually use behavioral tests such as righting reflex and roller running to evaluate the degree of drunkenness in mice. These behavioral tests are not very sensitive and can only determine the degree of severe drunkenness, such as coma. However, the overall performance of mice in the hangover state is often better than that in the drunken state, and the behavioral activity ability has been partially restored. At this time, more sensitive and sophisticated behavioral experiments are needed to accurately detect it.

[0150] The mouse open field test and rotarod test can better evaluate the mouse's exploration, general activity, movement and coordination ability, and are ideal behavioral tests for evaluating the overall degree of hangover in mice.

[0151] The present invention establishes a precise detection method for the degree of hangover in mice based on the spatiotemporal dynamic changes of ethanol metabolism. Using indicators related to ethanol drunkenness, it is found that mice have a hangover after 6 hours of ethanol metabolism ends after 2.5g / kg alcohol is intragastrically administered, which is an ideal hangover detection point. A mathematical analysis method that integrates various indicators is established to quantitatively calculate the degree of hangover of ethanol. The model is used to detect the degree of hangover of physiological saline and a certain liquor sample, and the reliability and sensitivity of the model are further verified. Therefore, the model of the present invention can be used for the precise detection of the degree of ethanol hangover in mice.

[0152] The patent of this invention discloses a method and mathematical analysis model for accurately detecting the degree of hangover based on the spatiotemporal changes of ethanol metabolism in mice, which can accurately detect the degree of hangover in mice in a comprehensive, objective and quantitative manner, and has high credibility and sensitivity.

[0153] The present invention not only has a wide range of applications, but also has great application potential. First, it can be used to accurately detect the degree of hangover of conventional wine samples; second, this patent can be used to improve or develop new brewing processes, such as evaluating the impact of various flavor substances on the degree of hangover, so as to target the flavor substances to improve the process; in addition, it can also be used to evaluate the efficacy of various alcohol remedies and test their actual efficacy in reducing the degree of ethanol hangover. Finally, through large-scale experiments, relevant industry standards can be formulated, and the degree of hangover can be included in the wine quality evaluation standards, which will promote the development of the entire wine industry, provide consumers with high-quality wine, and actively promote the internationalization of my country's liquor.

[0154] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A method for constructing a mathematical analysis model for accurate detection of hangover degree, characterized in that: Includes steps: (S1) Provide two groups of mice with the same origin and baseline conditions and raise them under the same environmental conditions; (S2) administering alcohol orally to one group of mice, and administering ethanol orally to another group of mice; (S3) performing an open field test on the mice in the alcohol gavage group and the mice in the ethanol gavage group, respectively, and recording the probability P1 that the distance moved by the mice in the alcohol gavage group in the open field is smaller than the distance moved by the mice in the ethanol gavage group in the open field; (S4) performing a rotating rod test on the mice in the alcohol gavage group and the mice in the ethanol gavage group, respectively, and recording the probability P2 that the rotating rod time of the mice in the alcohol gavage group is shorter than that of the mice in the ethanol gavage group; (S5) respectively testing the blood ethanol concentration of the mice in the wine gavage group and the ethanol gavage group, and recording the probability P3 that the blood ethanol concentration of the mice in the wine gavage group is greater than the blood ethanol concentration of the mice in the ethanol gavage group; and (S9) Based on each recorded probability, different weights W are assigned to each recorded probability according to the correlation between each detection index and the degree of hangover i ; Establish a mathematical analysis model for accurate detection of hangover degree, so as to obtain the hangover index (Hangover Index) P: Among them, step (S3), step (S4), and step (S5) can be performed in any order.

2. The method according to claim 1, characterized in that The method may also optionally include the following steps: (S6) respectively detecting the blood acetaldehyde concentration of the mice in the alcohol gavage group and the mice in the ethanol gavage group, and recording the probability P4 that the blood acetaldehyde concentration of the mice in the alcohol gavage group is lower than the blood acetaldehyde concentration of the mice in the ethanol gavage group; (S7) respectively detecting the concentration of acetaldehyde dehydrogenase (ALDH) in the liver of the mice in the alcohol gavage group and the mice in the ethanol gavage group, and recording the probability P5 that the concentration of ALDH in the liver of the mice in the alcohol gavage group is lower than that of the mice in the ethanol gavage group; (S8) Detecting cerebellar gamma-aminobutyric acid (GABA) in the mice in the alcohol gavage group and the mice in the ethanol gavage group, respectively, and recording the probability P6 that the cerebellar GABA level in the mice in the alcohol gavage group is greater than the cerebellar GABA level in the mice in the ethanol gavage group.

3. The method according to claim 1, characterized in that The experiments and detections of the method are carried out 6-8 hours after mice are gavaged with ethanol or alcohol.

4. The method according to claim 1, characterized in that In the intragastric administration of the wine and ethanol, the ethanol content of the wine and ethanol is the same.

5. An evaluation system for detecting the degree of hangover, characterized in that: The system comprises: (M1) Input module: The input module is configured to input open field test data, rotating rod test data and blood alcohol concentration data of the subject to be tested; (M2) Evaluation module: The evaluation module is configured to evaluate the open field test data, the rotating rod test data, and the blood ethanol concentration data based on the mathematical analysis model for accurate detection of the degree of hangover, so as to obtain an evaluation result of the degree of hangover of the subject to be tested; wherein the mathematical analysis model for accurate detection of the degree of hangover is a mathematical analysis model for accurate detection of the degree of hangover constructed by the construction method according to claim 1; the model formula is: and (M3) Output module: The output module is configured to output the evaluation result.

6. The system according to claim 5, characterized in that The greater the evaluation result value of the evaluation is, the higher the degree of hangover is; conversely, the smaller the evaluation result value of the evaluation is, the lower the degree of hangover is.

7. The system according to claim 5, characterized in that The alcoholic beverages include those selected from the group consisting of: white wine, yellow wine, fruit wine, beer, rice wine, medicinal wine, or a combination thereof.

8. The system according to claim 5, characterized in that The data input by the input module also includes data selected from the following group: blood acetaldehyde concentration, cerebellar GABA level, liver ALDH concentration, or a combination thereof.

9. An evaluation method, characterized in that Includes steps: (a) providing the open field test data, rotarod test data and blood ethanol concentration data of the test subject; (b) inputting the open field test data, the rotarod test data and the blood ethanol concentration data into the mathematical analysis model for accurate detection of hangover degree constructed by the construction method of claim 1, wherein the model formula is: (c) using the mathematical analysis model for accurate detection of hangover degree to evaluate the open field test data, the rotarod test data and the blood ethanol concentration data, thereby obtaining the hangover degree of the subject to be tested; and (d) Output the evaluation results.

10. The evaluation method according to claim 9, characterized in that: The greater the evaluation result value of the evaluation is, the higher the degree of hangover is; conversely, the smaller the evaluation result value is, the lower the degree of hangover is.

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