CN115537458A - Application of fecal flora detection-based method in evaluation of nutritional metabolism level of vegetarian pregnant women - Google Patents
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Abstract
The invention belongs to the field of biomedicine, and particularly relates to application of fecal flora detection in evaluation of nutrition metabolism level of vegetarian pregnant women.
Description
Technical Field
The invention belongs to the field of biological medicines, and particularly relates to application of fecal flora detection in evaluation of nutritional metabolism level of vegetarian pregnant women.
Background
During pregnancy, maternal intestinal flora is associated with the metabolic state of the mother and the infant. However, no technology exists for monitoring the nutrition of mothers who eat vegetarian diets during pregnancy. The existing hospital diagnosis and treatment technology focuses on simple indexes such as anemia, vitamin A and vitamin D deficiency of pregnant women, and further refined evaluation on individual conditions is difficult to achieve.
The Chinese pregnant woman dietary guideline (2016) recommends that: the basic principle of the general population dietary guidelines includes various foods, mainly cereals, balanced eating, healthy body constitution, more vegetables, fruits, milk and soybeans, proper amount of fish, poultry, eggs and lean meat, less salt and oil, and sugar control and wine limitation.
The reasonable diet principle of the pregnant women should supplement the following five aspects on the basis of the above diet guideline principle:
1. folic acid is supplemented, and iodine salt is selected for eating foods rich in iron; folic acid replenisher 400 mug/d should be taken orally throughout pregnancy, and green vegetables should be taken daily; in the middle and late pregnancy, 20-50 g of red meat should be added every day, and the viscera or blood of the animal is eaten 1-2 times every week; besides the daily intake of iodized salt, pregnant women should often eat marine foods such as kelp and laver.
2. For patients with severe vomiting, a small amount of multiple meals can be taken, and food containing necessary amount of carbohydrate is guaranteed to be taken; the patients without obvious early pregnancy reaction in the early pregnancy should continue to keep the balance diet before pregnancy; pregnant women with obvious pregnancy and vomiting or poor appetite do not need to over-stress balanced diet; during pregnancy, at least 130g of carbohydrate, preferably digestible cereal grain food, must be taken per day; patients with poor appetite or severe vomiting need to seek physician help.
3. The intake of milk, fish, poultry, eggs and lean meat is properly increased in the middle and late pregnancy; the total intake of the milk and the products thereof in the middle and late pregnancy reaches 300-500 g/d; in the middle and late pregnancy, 150-250 g of fish, poultry, eggs and lean meat are added every day; preferably, deep-sea fish is eaten for 2-3 times per week.
4. Proper physical activity is performed, and proper weight is maintained in the pregnancy period; proper weight gain during pregnancy is beneficial to obtaining good pregnancy outcome, and attention should be paid to physical quality monitoring and management; the body mass of the early pregnancy does not change greatly, and can be measured for 1 time per month, and the body mass of the middle and late pregnancy should be measured for 1 time per week; healthy pregnant women should perform moderate physical activity for not less than 30 minutes per day.
5. The product is not smoking and drinking, and is enjoyable to breed new life, and is actively prepared for breast feeding.
However, it may be difficult to meet all the guidelines due to different family dietary habits or individual differences, especially for pregnant women who are vegetarian for a long time, whether their dietary structure is reasonable, and whether they can meet the nutritional needs of the fetus and mother is difficult to evaluate with guidelines. Dietary differences contribute to differences in the level of deep metabolism in individuals via the maternal intestinal flora and are therefore of central importance for the assessment of nutritional status of the pregnant woman.
Comprehensive and dynamic means for assessing nutrition, including dietary surveys, i.e., dietary habits during pregnancy; physical examination including measurement of body weight, sebum thickness, waist circumference, hip circumference, etc.; laboratory examination including blood biochemical level, blood routine level, blood vitamins and trace elements, and comprehensive and dynamic determination of nutritional status during pregnancy based on the above indexes. However, the macro-representation of the general population is investigated, and the individual nutrition difference of vegetarian pregnant women caused by the difference of different nutrition structures is ignored.
Therefore, the method has great significance for carrying out refined evaluation on the nutrition metabolism level of the pregnant women so as to obtain more accurate nutrition state of the pregnant women.
Disclosure of Invention
Aiming at the problem that nutrition state assessment specially aiming at vegetarian pregnant women is lacked in the prior art, the scheme aims to provide application of fecal flora detection in assessing nutrition metabolism levels of the vegetarian pregnant women, and by detecting relative abundance of fecal flora of the vegetarian pregnant women, individual and refined assessment of the nutrition metabolism states of the vegetarian pregnant women is realized, and the nutrition metabolism levels of the vegetarian pregnant women can be more accurately reflected.
Based on the above purpose, the technical scheme adopted by the invention is as follows:
in a first aspect, the invention provides a use of a fecal flora based assay for assessing nutritional metabolic levels of a vegetarian pregnant woman, the fecal flora assay comprising detecting the relative abundance of fecal flora associated with the nutritional status of the vegetarian pregnant woman; the fecal flora comprises: collinsella, holdemania, unlasicified S247, eubacterium, roseburia, unlasicified Lachnospiraceae, clostridium and Acidococcus.
The invention screens out 8 fecal bacteria highly related to the nutrition metabolism level of vegetarian pregnant women for the first time, and the abundance of the bacteria in the feces of the vegetarian pregnant women is detected to further evaluate the nutrition metabolism level of the vegetarian pregnant women, thereby having important significance for the diet guidance of the vegetarian pregnant women.
In a second aspect, the present invention provides a model for assessing the nutritional metabolic level of a vegetarian diet pregnant woman based on the abundance of fecal flora, the model being constructed from the relative abundance of fecal flora associated with the nutritional status of the vegetarian pregnant woman, the fecal flora comprising: collinsella, holdemania, unlasicified S247, eubacterium, roseburia, unlasicified Lachnospiraceae, clostridium and Acidococcus.
Preferably, the model score formula is as follows:
the model score =3.3082-17.0407 × collinesella relative abundance value-47.7252 × Holdemania relative abundance value-64042.2145 × Unclassified S247 relative abundance value-294.1574 × Eubacterium relative abundance value-35.7225 × Roseburia relative abundance value-4042.2451 × Unclassified Lachnospiraceae relative abundance value-82.1704 × Clostridium relative abundance value-94.5717 × Acidaminococcus relative abundance value.
Preferably, when the model score is above 0.759 or below 0.532, it is suggested that the nutritional level of the vegetarian pregnant woman is to be adjusted.
The model for evaluating the nutrition metabolism level of the vegetarian diet pregnant women based on the abundance of the fecal flora has evaluation accuracy of not less than 77% on the nutrition metabolism level of the vegetarian diet pregnant women, and has important significance on diet guidance of the vegetarian diet pregnant women.
In a third aspect, the invention provides a construction method of the model for evaluating the nutrition metabolism level of the vegetarian diet pregnant woman based on the abundance of the fecal flora, which comprises the following steps:
collecting feces samples of vegetarian pregnant women and omnivorous pregnant women, and comprehensively scoring the nutritional metabolism level of the vegetarian pregnant women and the omnivorous pregnant women;
constructing a gene library of the pregnant woman fecal flora, and obtaining fecal flora OTU clustering and species annotation through sequencing to obtain the relative abundance of the fecal flora;
screening out fecal flora associated with the nutritional metabolism level of the vegetarian pregnant women by comparing the relative abundance of the fecal flora of the omnivorous pregnant women and the vegetarian pregnant women and the comprehensive score of the nutritional metabolism level;
and constructing a model for evaluating the nutrition metabolism level of the vegetarian diet pregnant women based on screening out fecal flora associated with the nutrition metabolism level of the vegetarian diet pregnant women.
Preferably, the step of screening the fecal flora associated with the nutritional metabolism level of the vegetarian pregnant women by comparing the fecal flora abundance and the nutritional metabolism level comprehensive score of the omnivorous pregnant women and the vegetarian pregnant women is as follows:
the relative abundance of the fecal flora of the two groups of people, namely the omnivorous pregnant women and the vegetarian pregnant women, and the corresponding nutrition metabolism level are respectively regressed through a random forest algorithm, importance ranking is carried out according to the relative contribution of each genus to the model prediction accuracy, the relationship between the number of the genus and the model error in the model is evaluated by a cross-folding cross-validation method, the genus is selected according to the importance ranking, and the fecal flora associated with the nutrition metabolism level of the vegetarian pregnant women is screened out.
Preferably, the selected genera of gut flora that are highly correlated with the level of vegetative metabolism in vegetarian pregnant women include Collinsella, holdemania, unlasied S247, eubacterium, roseburia, unlasied Lachnospiraceae, clostridium and Acidaminococcus.
In a fourth aspect, the invention provides application of the model for evaluating the nutrition metabolism level of the vegetarian diet pregnant women based on the abundance of the intestinal flora in preparing a nutrition state evaluation kit for the vegetarian diet pregnant women.
Compared with the prior art, the invention has the following beneficial effects:
the method screens fecal flora highly related to the nutrient metabolism level of vegetarian pregnant women for the first time, evaluates the nutritional state of the vegetarian pregnant women by detecting the combination abundance of specific intestinal microbiota on the basis of the flora abundance characteristics and contributes to the fine evaluation of the overall nutritional state of the vegetarian pregnant women so as to obtain more accurate nutritional state of the pregnant women and further provide reasonable guidance opinions, and has important significance on the nutrition monitoring and guidance of the pregnant women during pregnancy.
Drawings
FIG. 1 is a flow of the present case for recruiting subjects and exclusion criteria;
FIG. 2 is a graph showing the difference in intestinal flora between a vegetarian diet pregnant woman and a non-vegetarian diet pregnant woman;
FIG. 3 is a Lefse assay screening for differential genera;
FIG. 4 is a dissimilarity genus statistical information;
FIG. 5 is a ROC curve of a model for assessing the nutritional metabolic status of vegetarian diet pregnant women based on abundance of gut flora;
FIG. 6 is a ROC curve of a model for assessing the nutritional metabolic state of vegetarian diet pregnant women based on gut flora abundance versus test set analysis;
FIG. 7 is a graph showing the correlation between the nutrient metabolism and the intestinal flora of vegetarian and omnivorous pregnant women.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to the following examples. It will be understood by those skilled in the art that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The test methods used in the examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are commercially available unless otherwise specified.
Example 1
The application of the fecal flora-based detection in evaluating the nutrition of vegetarian pregnancy women comprises the following experimental steps:
(1) Sample and data acquisition
The experimental sample and data collection flow chart of the invention is shown in fig. 1, and pregnant women of 25-35 years of age and 3-4 months of gestation are recruited to take a frequency questionnaire and provide stool samples thereof. The study was approved by the fifth hospital affiliated with Guangzhou medical university, with all participants providing informed written consent. 40 pregnant women were recruited as a training set for the study, 12 of which were determined to be vegetarians and 28 of which were omnivores according to the questionnaire. The Body Mass Index (BMI) of the pregnant women at baseline (baseline at 12 weeks gestation), total energy intake at the time of the enrollment study, and future development of gestational diabetes were matched. Each woman completed the victoria cancer committee food frequency questionnaire (DQES V2.0 edition), directing dietary information including only from the onset of pregnancy. Researchers have performed macro-nutrient and fatty acid content analysis on the dietary information provided by pregnant women. Pregnant women also provided their own stool samples, which were refrigerated to-80 ℃ for storage prior to stool DNA isolation. In addition, fasting blood samples were collected, anthropometric data and medical and obstetric history were collected for all participants. The nutrition scoring of the pregnant women is based on the growth curve standard, healthy pregnant women generally accord with the growth curve standard, vegetarian pregnant women do not accord with the growth curve standard, but the vegetarian pregnant women do not necessarily mean malnutrition of the vegetarian pregnant women, so the nutrition scoring method analyzes and brings the faecal flora into the nutrition metabolism evaluation index of the vegetarian pregnant women.
(2) Data analysis
Establishing a gene library of fecal flora of pregnant women, obtaining OTU clustering and species annotation of intestinal flora by 16S rRNA sequencing to obtain species diversity and species abundance in an intestinal flora sample, comparing the flora difference of a heteroeating pregnant woman and a long-term vegetarian pregnant woman, and screening the differential flora by Lefse analysis, wherein the results are shown in figure 1 and figure 2, and the species with significant difference in abundance in different groups, namely the biomarkers with statistical difference are shown. The length of the histogram represents the magnitude of the effect of the significantly different species. Vegetarians were compared with other omnivores and screened for differentially by Lefse analysis.
(3) Results of the experiment
Based on the data analysis results, 12 vegetarians and 28 omnivores samples were used as training sets in a random forest (R language randomForest 4.6 14package) to analyze the nutritional metabolism level of the pregnant women, as shown in fig. 5. Regression analysis is carried out on the relative abundance of the genera of the two groups of people and the comprehensive scores of the corresponding nutrition metabolism levels, the relative contribution of each genus to the model prediction accuracy is ranked according to importance, the relation between the number of the genera in the model and the model error is evaluated by using a ten-fold cross-validation method, the genera are selected according to the importance ranking, finally, 8 remaining genera are used for constructing the random forest regression model of the invention, and 8 genera highly related to the food preference and the nutrition state are determined (as shown in fig. 2, fig. 3 and fig. 4): collinsella, holdemania, unlasicified S247, eubacterium, roseburia, unlasicified Lachnospiraceae, clostridium and Acidococcus.
(4) Construction of random forest regression model
Based on the determined 8 most important genera closely related to the nutrient metabolism level of the vegetarian pregnant women, an evaluation model of the nutrient metabolism level of the vegetarian pregnant women is constructed by carrying out regression analysis on the relative abundance values of the 8 genera of the flora and the corresponding comprehensive scores of the nutrient level, and the fitting model AUC reaches 0.924. The evaluation model formula calculates a score of =3.3082-17.0407 × Collinsella relative abundance value-47.7252 × Holdemania relative abundance value-64042.2145 × Uncsified S247 relative abundance value-294.1574 × Eubacterium relative abundance value-35.7225 × Roseburia relative abundance value-4042.1 × Uncsaid Lachnospiraceae relative abundance value-82.1704 × Clostridia relative abundance value-94.5717 × Acidaminococcus relative abundance value; a formula score of more than 0.759 or less than 0.532 indicates that the patient needs careful attention and adjustments to the nutritional levels, as illustrated in fig. 7 and table 1.
(5) Verification of random forest regression model
According to the training set collection specification, 20 feces samples of each of the pregnant woman feces with the miscellaneous food and the vegetarian food are independently collected to serve as a test set, the performance of the constructed random forest regression model is tested, the ROC curve for nutrition evaluation of the vegetarian pregnant woman is shown in figure 6 by referring to the judgment standard, and the accuracy of the random forest regression model for nutrition level evaluation of the vegetarian pregnant woman can reach 0.77 (area AUC under the curve).
(6) Application of random forest regression model
Through 16S rRNA sequencing of a training set and a verification set, intestinal flora clustering and species annotation are obtained, species diversity and species abundance in intestinal flora samples are obtained, network analysis is carried out, as shown in figure 7, figure 7 is a biological synthesis function prediction analysis result, and the result shows that the microbiota related to the biological synthesis ways of fatty acid, lipid, folic acid and the like are more obvious in vegetarians.
The results show that the microbiota related to the biosynthesis pathways of fatty acid, lipid, folic acid and the like are more remarkable for pregnant women who eat vegetarians for a long time compared with ordinary miscellaneous foods. We further summarize the results of the analysis as shown in table 1, where table 1 shows the correlation between specific genera and related nutritional indicators, and predict whether the diet of vegetarian pregnant women is nutritionally balanced by means of a model, and then the clinician judges whether the dietary structure needs to be adjusted by determining the relative abundance of specific genera in vegetarian pregnant women, for example, by evaluating one of the vegetarian pregnant women in the verification set, and finds that the calculated value of the model is less than 0.532, and determines that the nutritional structure needs to be adjusted, and according to the further analysis of the flora abundance, the abundance of the genus Unclassified Lachnospiraceae in the pregnant women is lower than that of other vegetarians, and referring to table 1, the clinician needs to adjust the fibrous dietary structure of the pregnant women accordingly.
In conclusion, the metabolism of vegetarian pregnant women on different dietary structures is carefully evaluated through the analysis of the metabolic pathways of intestinal flora, and table 1 can be used for supplementing relevant nutrition for pregnant women with different dietary habits guided by clinical nutrition. The obtained bacteria are subjected to biosynthesis function prediction analysis, and guidance opinions are provided for nutrition supplement of vegetarian pregnant women.
TABLE 1 correlation of specific genera with clinical characteristics and dietary intake
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (8)
1. Use of a fecal flora based assay for assessing nutritional metabolic levels in vegetarian pregnant women, wherein the fecal flora assay comprises measuring the relative abundance of fecal flora associated with the nutritional status of a vegetarian pregnant woman; the fecal flora comprises: collinsella, holdemania, unlasicified S247, eubacterium, roseburia, unlasicified Lachnospiraceae, clostridium and Acidococcus.
2. A model for assessing the nutritional metabolic level of vegetarian diet pregnant women based on the abundance of fecal flora, the model constructed from the relative abundance of fecal flora associated with the nutritional status of vegetarian pregnant women, the fecal flora comprising: collinsella, holdemania, unlasicified S247, eubacterium, roseburia, unlasicified Lachnospiraceae, clostridium and Acidococcus.
3. The model for assessing the nutritional metabolic level of a vegetarian diet pregnant woman based on fecal flora abundance according to claim 2 wherein the model score is formulated as follows:
the model score is =3.3082-17.0407 × collinesella relative abundance-47.7252 × Holdemania relative abundance-64042.2145 × outstanding S247 relative abundance-294.1574 × Eubacterium relative abundance-35.7225 × Roseburia relative abundance-4042.2451 × outstanding Lachnospiraceae relative abundance-82.1704 × Clostridium relative abundance-94.5717 × acidamicoccus relative abundance.
4. The model of claim 3 for assessing the nutritional metabolic level of a vegetarian diet pregnant woman based on fecal flora abundance, wherein a model score above 0.759 or below 0.532 indicates that the nutritional level of the vegetarian pregnant woman is to be adjusted.
5. The method for constructing a model for evaluating the nutritional metabolism level of a vegetarian diet pregnant woman based on the abundance of fecal flora according to any one of claims 2 to 4, comprising the steps of:
collecting feces samples of vegetarian pregnant women and omnivorous pregnant women, and comprehensively scoring the nutritional metabolism level of the vegetarian pregnant women and the omnivorous pregnant women;
constructing a gene library of the fecal flora of the pregnant woman, and obtaining the OTU cluster and species annotation of the fecal flora through sequencing to obtain the relative abundance of the fecal flora;
screening out fecal flora associated with the nutritional metabolism level of vegetarian pregnant women by comparing the relative abundance of the fecal flora of the omnivorous pregnant women and the vegetarian pregnant women and the comprehensive score of the nutritional metabolism level;
and constructing a model for evaluating the nutritional metabolism level of the vegetarian diet pregnant woman based on the screened fecal flora associated with the nutritional metabolism level of the vegetarian diet pregnant woman.
6. The construction method of claim 5, wherein the step of screening out the fecal flora associated with the nutritional metabolism level of the vegetarian pregnant women by comparing the fecal flora abundance and the nutritional metabolism level comprehensive scores of the omnivorous pregnant women and the vegetarian pregnant women comprises the following steps:
the relative abundance of the fecal flora of the two groups of people, namely the omnivorous pregnant women and the vegetarian pregnant women, and the corresponding nutrition metabolism level are respectively regressed through a random forest algorithm, importance ranking is carried out according to the relative contribution of each genus to the model prediction accuracy, the relationship between the number of the genus and the model error in the model is evaluated by a cross-folding cross-validation method, the genus is selected according to the importance ranking, and the fecal flora associated with the nutrition metabolism level of the vegetarian pregnant women is screened out.
7. The construction method of claim 6, wherein the selected intestinal flora associated with the nutritional metabolic level of vegetarian pregnant women comprises Collinsella, holdemania, uncelassiad S247, eubacterium, roseburia, uncelassiad Lachnospiraceae, clostridium and Acidaminococcus.
8. Use of the model for assessing the nutritional metabolic level of a vegetarian diet pregnant woman based on the abundance of fecal flora as claimed in any one of claims 2 to 4 in the preparation of a kit for assessing the nutritional status of a vegetarian pregnant woman.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114292930A (en) * | 2021-12-14 | 2022-04-08 | 上海交通大学医学院附属瑞金医院 | Application of fecal flora detection in children inflammatory bowel disease |
| CN117690476A (en) * | 2023-12-11 | 2024-03-12 | 广东医科大学附属医院 | Prediction method for evaluating contribution rate of intestinal flora to arsenic metabolism by using zebra fish model |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114292930A (en) * | 2021-12-14 | 2022-04-08 | 上海交通大学医学院附属瑞金医院 | Application of fecal flora detection in children inflammatory bowel disease |
| CN114292930B (en) * | 2021-12-14 | 2023-12-26 | 上海交通大学医学院附属瑞金医院 | Application of fecal flora-based detection in children inflammatory bowel disease |
| CN117690476A (en) * | 2023-12-11 | 2024-03-12 | 广东医科大学附属医院 | Prediction method for evaluating contribution rate of intestinal flora to arsenic metabolism by using zebra fish model |
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