Application of a composition in the preparation of food or medicine that reduces blood lead levels

Abstract

This invention discloses the application of a composition in the preparation of foods or medicines that reduce blood lead levels, belonging to the field of food or pharmaceutical applications. The composition is for improving intestinal microecology, comprising inulin, galactooligosaccharides, polydextrose, and water-insoluble dietary fiber, wherein the weight ratio of inulin, galactooligosaccharides, polydextrose, and water-insoluble dietary fiber is 25:25:20:30, and the composition is taken immediately after being dissolved in warm water before meals. This invention, through the rational combination of water-insoluble dietary fiber and functional polysaccharides, adjusts the release rate of nutrients in the intestine, slows down digestion, absorbs and eliminates toxic substances from food, reduces intestinal permeability, and simultaneously possesses the functions of water absorption and swelling, gradient adhesion, mechanical isolation, mesh adsorption, ion exchange, and microbial regulation, providing a favorable environment and food for the growth of intestinal microorganisms, and effectively reducing blood endotoxins and blood lead levels, enhancing immunity, and preventing chronic diseases.

Description

[0001] This application is a divisional application with application number 201880033374.X, application date May 22, 2018, priority number 201710367848.1, priority date May 23, 2017, and the invention title at the time of application was "Anti-inflammatory composition and food and application for improving intestinal function and microecology of nutritional metabolism". Technical Field

[0002] This invention relates to the field of food or pharmaceuticals that reduce blood lead levels, and in particular to the use of a composition in the preparation of food or pharmaceuticals that reduce blood lead levels. Background Technology

[0003] Since World War II, with infectious diseases controlled by vaccines and antibiotics respectively, chronic diseases now account for over 80% of all human deaths. The causes of chronic diseases include the gut-related theory and lifestyle theory. The core of the gut-related theory is that changes in diet, chemical substances, and antibiotics damage the intestinal barrier, leading to increased inflammatory factors in the blood, metabolic disorders, and immune dysfunction. These cumulative effects induce chronic diseases. In other words, improving intestinal barrier function can effectively prevent the occurrence of gut-related chronic diseases.

[0004] The intestinal barrier is divided into biological, chemical, mechanical, and immune barriers. The chemical barrier comprises various chemical substances secreted by glands in the gastrointestinal tract, which are relatively difficult to detect non-invasively and target for intervention. The mechanical barrier is characterized by intestinal mucus thickness, goblet cell count, villus length, crypt depth, and intestinal permeability. Measuring intestinal permeability is particularly convenient, allowing for easy detection, research, and development of intervention products. Like the mechanical barrier, the immune barrier's improvement can be easily monitored through the measurement of blood immune parameters. The biological barrier, namely the diversity of the gut microbiota, lacks a universally accepted standard for gut microbiota health. It is generally believed that a richer gut microbiota diversity, higher levels of lactic acid-producing bacteria, and greater production of short-chain fatty acids indicate a healthier gut. The essential correlation between gut microbiota and human health and disease is increasingly being proven. Apart from pathogenic microorganisms, there are no universally accepted sufficient factors to cause disease. However, when the intestinal mechanical barrier is damaged, increased intestinal permeability, coupled with a deteriorating gut microbiota, can produce a cumulative effect, leading to increased blood inflammatory factors, immune disorders, and metabolic disturbances—a recognized sufficient factor in inducing chronic diseases.

[0005] Regarding anti-inflammation and immunity, the body's immune balance is crucial for maintaining human health. Modern populations are often in a "sub-healthy state," where their immune systems are frequently subjected to various stresses or functional abnormalities, leading to inflammatory responses. This inflammatory response lies between the basal homeostatic state and the classic inflammatory response, and is termed low-grade chronic inflammation or para-inflammation (Medzhitov 2008). Cytokines are a class of small-molecule proteins with broad biological activity synthesized and secreted by immune cells or certain non-immune cells in response to external stimuli. They mainly include interleukins (IL), tumor necrosis factor (TNF), interferon (IFN), chemokines, and colony-stimulating factors (CSF), and can serve as biomarkers for the diagnosis of inflammation or disease. Reducing inflammatory factors, increasing anti-inflammatory factors, and improving chemokine levels are important requirements for reducing inflammation and maintaining overall health.

[0006] Many well-known conclusions exist regarding the relationship between nutritional metabolism and disease. Amidst widespread interest in glucose and lipid metabolism research, a scientific team led by Dr. Thomas J. Wang and Dr. Robert E. Gasten at Massachusetts General Hospital conducted a study on 2,422 individuals with normal blood glucose levels, following them for 12 years. Of these, 201 developed diabetes. The team used liquid chromatography-tandem mass spectrometry (LC-MS / MS) to analyze amino acids, amines, and other metabolites in a reference sample. They discovered a highly significant correlation between five branched-chain amino acids and aromatic amino acids and future diabetes: isoleucine, leucine, valine, tyrosine, and phenylalanine. These findings were replicated in an independent prospective cohort study. These findings highlight the potential importance of amino acid metabolism in the pathogenesis of diabetes and contribute to diabetes risk assessment and prevention.

[0007] Regarding the gut microbiota, current research suggests that the human body is a "superorganism" composed of human cells and all symbiotic microorganisms, and also a highly complex ecosystem. The number of microbial cells living inside and outside the human body is even 10 times the number of human cells themselves, numbering in the trillions. The adult gut is home to approximately 10¹⁴ bacteria, mainly colonizing the colon, forming a microbial ecosystem weighing about 1.5 kg, including 1000 to 1150 species of bacteria.

[0008] Some now believe that many health problems in the human body are caused by an imbalance in the gut microbiota, with the balance of the microecology being the determining factor. For example, gut microbiota is linked to obesity, and the mechanism by which gut microbiota imbalance leads to obesity mainly involves the following three aspects: some butyrate-producing bacteria can degrade polysaccharides in the diet, converting them into short-chain fatty acids for the body to absorb and utilize, thus increasing the host's ability to absorb energy from food; gut microbiota can also regulate the expression activity of genes related to energy storage tissues, mainly genes related to lipogenesis such as Fiaf and ChREBP / SREBP-1; a high-fat diet alters the structure of the gut microbiota, causing metabolic endotoxemia, and then inducing chronic systemic inflammation through the LPS / CD14 signaling pathway, leading to obesity, insulin resistance, etc. Other studies have shown that gut microbiota metabolites are associated with an increased incidence of major adverse cardiovascular events, and gut microbiota is correlated with colorectal cancer. Furthermore, some believe that toxins produced by gut microbiota are an important cause of aging and disease.

[0009] Based on new understandings of the gut microbiota, it is recognized as a crucial factor in the development of various chronic diseases. Current research reports that regulating the structure of the gut microbiota can influence the synthesis and secretion of various gastrointestinal hormones, such as increasing the concentrations of GLP-1 and GLP-2 in the colon and decreasing the concentration of dipeptidyl peptidase IV (DPP-4), thereby improving metabolic endotoxemia. Furthermore, α-glucosidase inhibitors can significantly increase the number of beneficial bacteria such as Bifidobacteria, improve glucose metabolism, and reduce inflammation in adipose tissue.

[0010] Since World War II, the food structure of most countries has undergone many changes. Since the 21st century, humanity has faced the enormous challenge of the "dietary fiber gap," which refers to the difficulty people face in consuming sufficient dietary fiber according to recommended standards in modern diets. This leads to a chronic state of low-grade inflammation, weakened immunity, and ultimately, various chronic diseases.

[0011] However, existing research and applications still have many problems: 1. The enormous challenge of the "dietary fiber gap" facing humanity in the 21st century has not been addressed, leading to obstructed nutrient metabolism, disruption of the gut microbiota, and consequently, a cumulative effect that induces various chronic diseases, contributing to the increasing prevalence of chronic diseases, which account for more than 80% of human deaths; 2. There is no effective anti-inflammatory combination that simultaneously improves the imbalance of nutrient metabolism, intestinal function, and gut microbiota. While adding functional polysaccharide combinations or ordinary polysaccharide combinations alone may have positive effects on the human body, it can also cause negative effects such as increased intestinal permeability and an excess or deficiency of certain beneficial bacteria; 3. There is a lack of research and application on the synergistic effects of complex fibers to compensate for the shortcomings and deficiencies of simple polysaccharide combinations. Supplementing with probiotics has the limitation of targeting the large intestine, and supplementing with prebiotic combinations carries the risk of increased intestinal osmotic pressure and gut microbiota imbalance; 4. Difficulty in choosing foods is a major obstacle to a balanced diet. There is a lack of effective and simple anti-inflammatory staple foods to facilitate primary prevention and fundamentally extend human healthy lifespan, rather than relying solely on secondary prevention and clinical medicine to extend the lifespan of humans with diseases.

[0012] Furthermore, lead is a heavy metal environmental pollutant with multi-systemic toxicity and is listed by the World Health Organization as one of the top ten public health chemicals of concern. Its toxicological mechanism is mainly based on its high affinity for sulfhydryl enzymes, which can interfere with the metabolism of essential metals such as calcium, iron, and zinc, inducing oxidative stress and mitochondrial dysfunction, ultimately leading to cell damage and apoptosis. Epidemiological studies have shown that even when blood lead levels are below 3.5 µg / dL, children's cognitive function still declines in a dose-dependent manner; for every 10 µg / L increase, IQ decreases by an average of 2–5 points, accompanied by attention deficit and behavioral abnormalities. Long-term low-dose exposure in adults is significantly associated with hypertension, atherosclerosis, and ischemic heart disease; in 2019, approximately 5.45 million cardiovascular deaths worldwide were attributed to lead exposure. In addition, lead can accumulate in bones (accounting for 95% of the total body weight), with a half-life of 10–30 years, and is released into the bloodstream during pregnancy, lactation, or osteoporosis, causing endogenous secondary exposure.

[0013] In summary, in recent years, correlation analysis techniques such as metabolomics, proteomics, metagenomics, immunomics, and microbiome have been increasingly applied to life science research. Research focuses on developing anti-inflammatory compositions that improve nutritional metabolism, intestinal function, and gut microbiota, eliminating pathogenic factors such as chronic inflammation, blood lead levels, and weakened immunity, improving sub-health conditions, and preventing chronic diseases. Developing anti-inflammatory foods that simultaneously meet human health needs and maintain gut microbiota is an important nutritional health issue. This invention, based on this understanding, develops a composition for use in the preparation of foods or pharmaceuticals that reduce blood lead levels, thereby improving gut microbiota while simultaneously reducing heavy metals in the blood. This will conveniently improve sub-health conditions and effectively prevent chronic diseases, making it a crucial research topic today. Summary of the Invention

[0014] The technical problem to be solved by the present invention is to provide an application of a composition in the preparation of food or medicine that reduces blood lead content, so as to achieve the purpose of reducing blood heavy metals while improving intestinal microecology.

[0015] To solve the above-mentioned technical problems, the present invention provides an application of a composition in the preparation of food or medicine that reduces blood lead content. The composition is a composition for improving intestinal microecology, comprising inulin, galactooligosaccharides, polydextrose, and water-insoluble dietary fiber, wherein the weight ratio of inulin, galactooligosaccharides, polydextrose, and water-insoluble dietary fiber is 25:25:20:30, and the composition is taken with warm water before meals and immediately after eating.

[0016] In a further improvement, the inulin is fructooligosaccharide, polyfructooligosaccharide, or a mixture of fructooligosaccharide and polyfructooligosaccharide; the water-insoluble dietary fiber is cellulose, hemicellulose, lignin, or a mixture thereof that is insoluble in water.

[0017] In a further improvement, the water-insoluble dietary fiber is wheat bran fiber extracted from wheat bran.

[0018] With this design, the present invention has at least the following advantages:

[0019] This invention's composition, through a rational combination of water-insoluble dietary fiber and functional polysaccharides, adjusts the release rate of nutrients in the intestines, slows down digestion, accelerates cholesterol excretion, absorbs and eliminates toxic substances from food, and particularly improves the metabolism of branched-chain amino acids and aromatic amino acids related to diabetes prevention. It also ensures the stability or reduction of blood trimethylamine oxidase biochemical indicators, reduces intestinal permeability, and simultaneously possesses water-absorbing swelling, gradient adhesion, mechanical isolation, mesh adsorption, ion exchange, and microbial regulation functions, providing a favorable environment and food for intestinal microbial growth and maintaining intestinal microbial homeostasis. The synergistic effect of the anti-inflammatory composition eliminates the negative effects of using functional polysaccharides alone, reduces blood endotoxins, blood lead, and inflammatory factors, enhances cellular metabolism and immune capacity, eliminates pathogenic factors such as chronic inflammation and weakened immunity, and effectively prevents the occurrence of chronic diseases such as cardiovascular and cerebrovascular diseases and diabetes. This invention's composition can effectively improve nutrient metabolism, reduce intestinal permeability, maintain the homeostatic environment of intestinal microorganisms, and prevent chronic diseases.

[0020] The food containing the above-mentioned composition is an indispensable staple food for people every day. It provides the human body with a balanced nutritional composition, improves physical condition, effectively prevents foodborne chronic diseases, and protects health. Attached Figure Description

[0021] The above is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0022] Figure 1 This is a graph showing the change in pH value of mouse intestinal contents in Application Example 1 of the present invention.

[0023] Figure 2 This is a graph showing the changes in DAO, a biochemical marker in mouse serum, in Example 1 of the present invention.

[0024] Figure 3 This is a graph showing the changes in the serum biochemical indicator TMAO in mice during Example 1 of the present invention.

[0025] Figure 4 This is a graph showing the changes in the mouse serum biochemical index SCFA in Example 1 of the present invention.

[0026] Figure 5 This is a diagram showing the changes in the height of the villi and crypt sections of the mouse small intestine in Example 1 of the present invention.

[0027] Figure 6 This is a diagram showing the composition of the gut microbiota at the phylum level in the blank group mice during 21 days in Application Example 1 of this invention.

[0028] Figure 7 This is a diagram showing the composition of the gut microbiota at the phylum level in experimental group A mice during 21 days in Example 1 of this invention.

[0029] Figure 8 This is a diagram showing the composition of the gut microbiota at the phylum level in experimental group B mice during 21 days in Example 1 of this invention.

[0030] Figure 9 This is a diagram showing the composition of the gut microbiota at the phylum level in experimental group C mice during 21 days in Example 1 of this invention.

[0031] Figure 10 This is a diagram showing the composition of the gut microbiota at the genus level in the four experimental groups of mice during 21 days in Example 1 of this invention.

[0032] Figure 11 This is a scatter plot of the statistical results of blood biochemical indicators differences among all trial participants after 45 days of consumption in Example 2 of this invention. Figure 1 Among them, 11A represents hemoglobin content data, 11B represents mean hemoglobin content data, 11C represents mean hemoglobin concentration data, 11D represents immune factor IgA data, 11E represents insulin data, and 11F represents triglyceride data.

[0033] Figure 12This is a scatter plot of the statistical results of blood biochemical indicators differences among all trial participants after 45 days of consumption in Example 2 of this invention. Figure 2 Among them, 12A represents uric acid data, 12B represents serum lead content data, 12C represents albumin data, 12D represents globulin data, 12E represents albumin-to-globulin ratio data, and 12F represents alanine aminotransferase (ALT) data.

[0034] Figure 13 This is a scatter plot of the statistical results of blood biochemical indicators differences among all trial participants after 45 days of consumption in Example 2 of this invention. Figure 3 Among them, 13A represents total bilirubin data, 13B represents indirect bilirubin data, 13C represents high-density lipoprotein data, 13D represents low-density lipoprotein data, 13E represents D-lactic acid data, and 13F represents glutamine data.

[0035] Figure 14 This is a scatter plot of the statistical results of amino acid index differences among all trial participants after 45 days of consumption in Example 2 of this invention. Figure 1 Among them, 14A represents leucine data, 14B represents valine data, 14C represents phenylalanine data, 14D represents isoleucine data, 14E represents tyrosine data, and 14F represents the sum of the first five amino acid data.

[0036] Figure 15 This is a scatter plot showing the sum of tyrosine and five amino acid data for the grouped population after 45 days of food consumption in Example 2 of this invention. In this plot, 15A represents tyrosine data and 15B represents the sum of the first five amino acid data.

[0037] Figure 16 This is a scatter plot of D-lactic acid / glutamine data after 45 days of testing in all participants in Application Example 2 of this invention. 16A represents D-lactic acid data and 16B represents glutamine data.

[0038] Figure 17 This is a scatter plot of data on five decreased cytokines in all trial participants after 45 days of consumption in Example 2 of this invention. Among them, 17A is the data of cytokine IL-12p70, 17B is the data of cytokine IL-13, 17C is the data of cytokine IL-2, 17D is the data of cytokine IL-23, and 17E is the data of cytokine RANTES.

[0039] Figure 18 This is a scatter plot of data on five rising cytokines in all trial participants after 45 days of consumption in Example 2 of this invention. Among them, 18A is the data of cytokine IL-1RA, 18B is the data of cytokine IL-7, 18C is the data of cytokine TNF-β, 18D is the data of cytokine MCP-1, and 18E is the data of cytokine MIP-1α.

[0040] Figure 19 This is a scatter plot of data on four rising cytokines in all trial participants after 45 days of consumption in Example 2 of this invention. Among them, 19A is the data of cytokine IP-10, 19B is the data of cytokine TNF-α, 19C is the data of cytokine GM-CSF, and 19D is the data of cytokine IL-18.

[0041] Figure 20 This is a scatter plot of data on four rising cytokines in all trial participants after 45 days of consumption in Example 2 of this invention. Among them, 20A is the data of cytokine IL-27, 20B is the data of cytokine IL-15, 20C is the data of cytokine IL-4, and 20D is the data of cytokine GRO-α. Detailed Implementation

[0042] The present invention relates to an anti-inflammatory composition that improves nutrient metabolism, reduces intestinal permeability, and improves intestinal microecology, comprising inulin, galactooligosaccharides, polydextrose, and water-insoluble dietary fiber, wherein the ratio of inulin, galactooligosaccharides, polydextrose, and water-insoluble dietary fiber is 10-30: 5-30: 10-40: 10-50.

[0043] Inulin of this invention refers to fructooligosaccharides, polyfructooligosaccharides, or a mixture of fructooligosaccharides and polyfructooligosaccharides; galactooligosaccharides (GOS) are functional oligosaccharides with natural properties, whose molecular structure generally consists of 1 to 7 galactose groups attached to galactose or glucose molecules, i.e., Gal-(Gal)n-Glc / Gal (n is 0-6); polydextrose is a water-soluble dietary fiber; and solkfloc refers to cellulose, hemicellulose, lignin, or a mixture thereof that are insoluble in water, and the particle size can be microcrystalline cellulose.

[0044] To make the anti-inflammatory composition that improves nutrient metabolism, reduces intestinal permeability, and improves intestinal microecology more convenient to act on the human body, the present invention also provides an anti-inflammatory food containing the above-mentioned anti-inflammatory composition, the anti-inflammatory food further comprising cereal powder, wherein the formulation ratio of the cereal powder to the anti-inflammatory composition of the present invention is 65-95:35-5.

[0045] Grain flour refers to wheat flour, rice (japonica rice, indica rice, glutinous rice) flour, corn flour, potato flour or combinations thereof. The balanced anti-inflammatory food forms formed by the grain flour and the above-mentioned combinations mainly include anti-inflammatory rice, flour, noodles (rolled noodles, dried noodles, semi-dried noodles, wet noodles), pasta, rice noodles, bread, bread dough or bread premix, etc., and are not limited to these forms.

[0046] The preferred formulation ratio of the cereal powder to the anti-inflammatory composition of the present invention is 75-90:25-10.

[0047] Of course, the anti-inflammatory food of the present invention may also include minerals, vitamins, and nutritional fortifiers and supplements such as xylooligosaccharides, L-arabinose, stachyose, and guar gum, and the scope of their addition complies with the requirements of the Codex Alimentarius Commission (CAC) General Principles for the Addition of Essential Nutrients to Foods.

[0048] The anti-inflammatory composition of this invention can improve nutritional metabolism. Water-insoluble dietary fiber slows down digestion and accelerates the excretion of cholesterol, endotoxins, and heavy metals, maintaining the intestinal microbial environment. It has functions such as water absorption and swelling, gradient adhesion, mechanical isolation, mesh adsorption, ion exchange, and microbial regulation. In this invention, inulin, galactooligosaccharides, and polydextrose can selectively stimulate the growth and activity of one or more beneficial bacteria, promoting their growth and reproduction while inhibiting the growth and activity of harmful bacteria. This balanced anti-inflammatory food can effectively maintain the homeostasis of intestinal microorganisms, reduce intestinal permeability, eliminate pathogenic factors such as chronic inflammation and immune disorders, effectively improve physical condition, and prevent chronic diseases.

[0049] Product Example 1: Anti-inflammatory Nutritional Rice

[0050] The cereal powder is a mixture of japonica rice flour and glutinous rice flour in a ratio of 80:20. The anti-inflammatory composition is a mixture of inulin, galactooligosaccharide, polydextrose, and microcrystalline cellulose in a ratio of 30:10:30:30. The ratio of cereal powder to anti-inflammatory composition is 85:15.

[0051] The preparation method of this anti-inflammatory and nutritious rice is as follows:

[0052] 1. Place each component of the anti-inflammatory composition in a mixer according to the specified proportions and mix thoroughly to prepare a premixed powder for later use;

[0053] 2. Mix japonica rice flour and glutinous rice flour in a ratio of 80:20 to form a rice flour mixture. Mix this rice flour mixture with the premixed powder obtained in step 1 in a ratio of 85:15. Add 27-29% water and stir to adjust the consistency to obtain an anti-inflammatory nutritious rice semi-finished product.

[0054] 3. The anti-inflammatory nutritious rice semi-finished product obtained in step 2 is extruded and granulated using a twin-screw extruder, then dried using a fluidized bed dryer, and then cooled and sieved to obtain the anti-inflammatory nutritious rice of this embodiment.

[0055] The finished anti-inflammatory nutritious rice of this embodiment resembles natural rice in appearance and has an attractive appearance. Its texture is delicate and smooth, without any grainy feel, and it does not become hard. Its functional characteristics include meeting the body's anti-inflammatory nutritional needs, having a glycemic index of less than 50, and improving constipation and the intestinal microecological environment.

[0056] Product Example 2: Anti-inflammatory Nutritional Flour

[0057] The cereal flour is made from ordinary wheat flour, and the anti-inflammatory composition is a mixture of inulin, galactooligosaccharides, polydextrose, and microcrystalline cellulose in a ratio of 30:5:40:25, with the ratio of cereal flour to anti-inflammatory composition being 80:20.

[0058] The preparation method of this anti-inflammatory nutritional flour is as follows:

[0059] 1. Place each component of the anti-inflammatory composition in a mixer according to the specified proportions and mix thoroughly to prepare a premixed powder for later use;

[0060] 2. Mix the regular flour with the premixed powder obtained in step 1 at a ratio of 80:20, stir thoroughly to adjust the consistency, and sift to obtain the anti-inflammatory nutritional flour of this embodiment.

[0061] The anti-inflammatory nutritional flour of this embodiment has a finished product that resembles ordinary flour, with a delicate texture and no grainy feel when used in various pasta dishes. Its functional characteristics include: meeting the body's anti-inflammatory nutritional needs, having a glycemic index of less than 50, and improving constipation and the intestinal microecological environment.

[0062] Product Example 3: Anti-inflammatory Nutritional Noodles

[0063] The cereal flour is made from ordinary wheat flour, and the anti-inflammatory composition is a mixture of inulin, galactooligosaccharides, polydextrose, and microcrystalline cellulose in a ratio of 10:30:10:50, with the ratio of cereal flour to anti-inflammatory composition being 65:35.

[0064] The preparation method of this anti-inflammatory and nutritious noodle is as follows:

[0065] 1. Place each component of the anti-inflammatory composition in a mixer according to the specified proportions and mix thoroughly to prepare a premixed powder for later use;

[0066] 2. Mix the all-purpose flour with the premixed powder obtained in step 1 at a ratio of 65:35, then add water and stir until the water content is about 45%. The water temperature should be 15-20℃. Stir for 10-15 minutes.

[0067] 3. Cook for 10-15 minutes at a temperature of about 25°C. After cooking, roll the dough, cut it into strips, dry it, and cool it with cold air to fix the strips. The moisture content will be reduced to 14%, thus obtaining the anti-inflammatory and nutritious noodles of this embodiment.

[0068] The anti-inflammatory and nutritious noodles in this embodiment have a delicate, smooth, and chewy texture. Their functional characteristics include: meeting the body's anti-inflammatory nutritional needs, having a glycemic index of less than 50, and improving constipation and the intestinal microecological environment.

[0069] Product Example 4: Anti-inflammatory Nutritional Bread

[0070] The cereal flour is medium-gluten flour, and the anti-inflammatory composition is a mixture of inulin, galactooligosaccharide, polydextrose, and microcrystalline cellulose in a ratio of 30:30:30:10, with the ratio of cereal flour to anti-inflammatory composition being 95:5.

[0071] The preparation method of this anti-inflammatory and nutritious bread is as follows:

[0072] 1. Place each component of the anti-inflammatory composition in a mixer according to the specified proportions and mix thoroughly to prepare a premixed powder for later use;

[0073] 2. Mix the all-purpose flour with the premixed powder obtained in step 1 at a ratio of 95:5, then add water and stir. Follow the traditional bread-making process: kneading, first fermentation, second dough preparation, second fermentation, shaping, molding, pre-baking processing, baking, and cooling to obtain the anti-inflammatory nutritional breadsticks of this embodiment. This step eliminates the need for ingredients such as white sugar and vegetable oil used in traditional bread-making methods due to the characteristics of the anti-inflammatory composition.

[0074] This embodiment of the anti-inflammatory nutritional bread has a delicate, smooth texture, a fluffy and soft structure, and is not prone to collapsing. Its functional characteristics include meeting the body's anti-inflammatory nutritional needs and improving constipation and the intestinal microecological environment.

[0075] The anti-inflammatory composition of this invention can improve nutritional metabolism. Water-insoluble dietary fiber slows down digestion and accelerates the excretion of cholesterol and heavy metals, maintaining the intestinal microbial environment. It has functions such as water absorption and swelling, gradient adhesion, mechanical isolation, mesh adsorption, ion exchange, and microbial regulation. In this invention, inulin, galactooligosaccharides, and polydextrose can selectively stimulate the growth and activity of one or more beneficial bacteria, promoting their growth and reproduction while inhibiting the growth and activity of harmful bacteria. This balanced anti-inflammatory food can effectively maintain the homeostasis of intestinal microorganisms, reduce intestinal permeability, eliminate pathogenic factors such as chronic inflammation and immune disorders, effectively improve physical condition, and prevent chronic diseases.

[0076] Application Example 1

[0077] This experiment categorized glycan compositions into three types: 1) functional glycan compositions; 2) ordinary glycan compositions; and 3) compositions containing multiple functional and ordinary glycans. These compositions were used to study the synergistic effects of different types of glycan compositions. The focus of this synergistic effect was twofold: firstly, changes in the abundance and living environment of suitable microorganisms; and secondly, changes in their impact on host physiological indicators. The specific experimental procedures are as follows:

[0078] 1. Materials and Equipment

[0079] 1.1 Reagents

[0080] DAO kit, SCFA kit, and TMAO kit were provided by Shanghai Enzyme-Link Biotechnology Co., Ltd.

[0081] 1.2 Laboratory Animals

[0082] SPF-grade 8-week-old BARBL / C mice were purchased from the Experimental Animal Center of the Academy of Military Medical Sciences.

[0083] 1.3 Experimental Feed

[0084] Four experimental feed groups were prepared according to the different classifications of the above-mentioned polysaccharide compositions:

[0085] The blank group O had 100% base food, which is the general feed for mice;

[0086] The experimental group A, consisting of a base material and functional polysaccharide composition, specifically comprises: 85% base material, 7.5% galactooligosaccharides, and 7.5% inulin.

[0087] The experimental group B, consisting of a base material and a common polysaccharide composition, specifically comprises: 85% base material, 7.5% insoluble dietary fiber (wheat bran extract), and 7.5% polydextrose.

[0088] Experimental group C, consisting of a base material, a functional polysaccharide composition, and a common polysaccharide composition, specifically comprises: 85% base material, 4% galactooligosaccharides, 3.5% inulin, 4% insoluble dietary fiber, and 3.5% polydextrose. All experimental feeds were provided by Beijing Ruiqianjing Technology Development Co., Ltd. This formulation is based on food application, assuming a daily average intake of 250-300 grams of staple food per person, and aims to provide a reference intake of 37-45 grams of complex dietary fiber.

[0089] 1.4 Main Instruments and Equipment

[0090] Electronic balance and pH meter, Mettler Toledo Instruments (Shanghai) Co., Ltd.; Gas chromatograph-mass spectrometer.

[0091] 2 Experimental Methods

[0092] 2.1 Experimental Design

[0093] Mice were randomly assigned to four groups: control group O, experimental group A, experimental group B, and experimental group C (8 mice per group). They were fed in an independent ventilation cage system with strict control of 12 hours of light / darkness and an ambient temperature of 22℃.

[0094] This experiment was conducted over four weeks, with week 1 being the acclimatization period and weeks 2, 3, and 4 being the experimental periods. During the acclimatization period, all four groups of mice were fed a blank diet (i.e., 100% basal food). During the experimental period, mice in the blank group (O) were fed a blank diet, while mice in the experimental groups were fed the corresponding diets for experimental groups A, B, and C, respectively. At the end of the second and third weeks of the experimental period, three mice were randomly selected, their eyeballs were enucleated to collect blood, and the mice were euthanized by dislocation. The small intestine was collected and stored in formalin solution for HE staining. Colonic contents samples were collected and stored at -80℃. pH, bacterial flora composition, etc., were analyzed. After blood collection, serum was separated, and various biochemical indicators in the serum, including diamine oxidase (DAO), trimethylamine oxide (TMAO), and short-chain fatty acids (SCFA), were measured using a kit.

[0095] 2.2 Determination of serum metabolic parameters

[0096] Blood was collected from mouse eyeballs and allowed to stand at room temperature for 1 hour. The collected serum was then centrifuged at 3000 rpm for 15 minutes and separated, and stored at -80°C. Serum biochemical markers, including DAO, TMAO, and SCFA, were measured using a kit.

[0097] 2.3 Determination of pH value of intestinal contents

[0098] Weigh out 100 mg of intestinal contents, add deionized water at a rate of 15 mL / g, disperse thoroughly, centrifuge at 13000g for 2 min, and measure the pH value of the supernatant using a pH meter.

[0099] 2.4 Measurement of intestinal contents metabolomics

[0100] Weigh 100 mg of sample and add a methanol:water:chloroform (3:1:1) solution. Vortex to disperse evenly, let stand for 12 hours, and centrifuge to collect the supernatant. Take 100 μl of the extract and add 20 μl of 0.2 mg / mL ribitol as an internal standard. Mix thoroughly and dry under N2 at 45 °C. Add 40 μL of 20 mg / mL methoxyamine hydrochloride solution to the dried extract, mix thoroughly, and react at 130 rpm for 90 min at 30 °C. After complete reaction, add 40 μL of BSTFA (containing 1% TMCS), mix thoroughly, and incubate in a gas bath at 37 °C for 30 min. Remove and place at room temperature for 120 min, then test at 4 °C.

[0101] GC-MS chromatographic conditions: GC conditions: Agilent 7890, tandem LECO Pegasus 4D TOF / MS detector;

[0102] Gerstel MPS injection system; Column: DB-5MS 30m × 250μm × 0.25mm

[0103] Temperature program: 70℃, hold for 1 min, increase to 280℃ at 5℃ / min, hold for 10 min.

[0104] Carrier gas: He; Flow rate: 1 mL / min; Injection volume: 1 μL; Split ratio: 1:2;

[0105] Mass spectrometry conditions:

[0106] MS conditions: EI source: mass scan range: 50-800 Da, scan rate 10 / s; injection port, transfer line and ion source temperatures: 250℃, 250℃ and 220℃ respectively.

[0107] Data processing

[0108] After peak alignment, deconvolution, and peak retrieval of each compound in the chromatogram using a Chroma TOF 4.50 workstation (identified using the built-in NIST mass spectrometry database and relevant standards), the original chromatogram was normalized using ribitol (internal standard). After integration (signal-to-noise ratio S / N > 100), the names of each chromatographic peak and metabolite, peak area, and other relevant data were finally obtained.

[0109] 2.5 Measurement of changes in small intestinal villus height and crypt depth

[0110] Image Pro-Plus 6.0 method for analyzing villus length, crypt depth and goblet cells in intestinal tissue: The line tool of IPP software was used to select images with relatively extended villus and intact intestinal villus to measure the villus length and crypt depth of each slice.

[0111] 2.6 Determination of 16SDNA in Intestinal Contents

[0112] Microbial DNA extraction was performed according to the instructions of the Intestinal Contents DNA Extraction Kit (Omega Bio-tek, Norcross, GA, US). The quality of the extracted DNA was assessed by 1% agarose gel electrophoresis. DNA samples were stored at -20º C. PCR amplification of the 16S v3-v4 hypervariable region of the bacterial 16S ribosomal RNA gene was performed using the Illumina MiSeq PE300 sequencing platform via high-throughput sequencing (amplification conditions: 95℃ for 2 min, 95℃ for 30 s, 55℃ for 30 s, 72℃ for 30 s, 25 cycles, followed by extension at 72℃ for 10 min). The primers used were: forward primer (515F) 5'-barcode-GTGCCAGCMGCCGCGG)-3', reverse primer (907R) 5'-CCGTCAATTCMTTTRAGTTT-3', and an 8 bp tag sequence was added to each sample. Each sample was amplified in triplicate. The reaction volume was 25 μL, including 2.5 μL of 10 × Pyrobest Buffer, 2 μL of 2.5 mM dNTPs, 1 μL each of forward and reverse primers (10 μM), 0.4 U of Pyrobest DNA Polymerase (TaKaRa), 15 ng of DNA template, and the remainder ddH2O. Amplicons were recovered from 2% agarose gels and purified according to the AxyPrep DNA Gel Extraction Kit instructions (Axygen Biosciences, Union City, CA, US), and then quantified using QuantiFluor™-ST (Promega, US). The purified amplicons were mixed in equal volumes and then sequenced at both ends (2 × 300 bp) according to the standard Illumina MiSeq sequencing platform protocol. The original FASTQ files were processed using QIIME (version 1.17) software, with the following processing criteria: (i) A sliding window of 10 bp was used. If the average quality value within the window was below 20, the last base was truncated from the beginning of the window, and sequences smaller than 50 bp were filtered out. (ii) Samples were distinguished based on tag sequences. The allowed number of mismatches for tag sequences was 0, and the allowed number of mismatches for primers was 2. Sequences containing ambiguous bases were removed. (iii) The overlap between sequences that were spliced ​​together must be at least 10 bp, and sequences that could not be spliced ​​were removed. Operational Units (OTUs) were generated using UPARSE (version 7.1, http: / / drive5.com / uparse / ) software, with a similarity of 97%. Chimeric sequences were then identified and removed using UCHIME software.For taxonomic alignment, the silva (SSU115) 16S ribosomal RNA database was used, and the algorithm was RDP Classifier (http: / / rdp.cme.msu.edu / ) with a confidence threshold of 70%.

[0113] 3. Results and Conclusions

[0114] 3.1 pH value of intestinal contents

[0115] See attached document Figure 1 As shown, in the control group O mice, the pH of the intestinal contents was 8.23 ​​and 8.17 at days 14 and 21, respectively. However, the pH of the intestinal contents in the experimental groups A, B, and C dropped sharply to around 6.0, with group C showing the most stable value. This is because bacteria break down oligosaccharides, producing short-chain fatty acids, which lowers the pH.

[0116] Intestinal pH is an important parameter of the intestinal microecology. The pH parameter changes significantly. As can be seen from this experiment, the feed formula of experimental group C is the most stable and ideal.

[0117] 3.2 Serum biochemical indicators in mice

[0118] Table 1. Changes in serum biochemical parameters of mice after dietary intervention in the experimental group.

[0119]

[0120] From Table 1 and Appendix Figures 2 to 4 It can be seen that the serum indicators of the experimental group mice were compared with those of the blank group as follows:

[0121] Experimental Group A: The DAO index showed unstable changes, the negative TMAO value increased, and the SCFA value did not change significantly.

[0122] Experimental Group B: The DAO index did not change significantly, the negative value of TMAO changed significantly over 21 days (became larger), and the SCFA fluctuated greatly.

[0123] Experimental group C: DAO, TMAO, and SCFA showed significant changes.

[0124] Since diamine oxidase (DAO) is an intracellular enzyme in intestinal mucosal cells, the level of DAO in the blood directly reflects the degree of damage to the intestinal mucosa and the level of intestinal permeability. It is generally believed that oligosaccharides selectively promote the proliferation of beneficial bacteria in the gut, thereby promoting host health. However, there are also reports that fructooligosaccharides increase intestinal permeability in rats, and at high doses, even increase the ectopic growth of Salmonella. Excessive consumption of oligosaccharides can also cause intestinal discomfort, such as borborygmus and bloating. Table 1 shows that at the end of the second week of the experiment, the DAO value of group A mice was the lowest, but it increased at the end of the third week. This may be because excessive functional oligosaccharide composition increased intestinal permeability in mice, leading to an increase in DAO value. Group C mice showed a gradual decrease, indicating that the synergistic effect of the formulation in group C compensated for the important deficiencies of the formulations in groups A and B, making it an ideal formulation.

[0125] Trimethylamine oxide (TMAO), a choline metabolite dependent on gut microbiota, plays an important role in the development and progression of cardiovascular diseases. Elevated host TMAO levels are significantly positively correlated with cardiovascular diseases. Gut microbiota use ingested nutrients such as lecithin, choline, and carnitine as carbon energy sources. Gut microbiota possess the ability to cleave trimethylamine (an enzyme not found in mammals), which breaks the C-N bonds in these nutrients, releasing trimethylamine as a metabolic waste product. This trimethylamine then enters the liver via the portal circulation, where it is oxidized by flavin monooxidase, primarily (FMO3), to generate TMAO.

[0126] At the end of the second and third weeks of the experiment, the values ​​of groups A and B were even higher than those of the blank group O, indicating a negative effect. This suggests that although the functional polysaccharide composition and the ordinary polysaccharide composition have an impact on changes in the intestinal microecology, they have a phased negative effect on the host. The serum TMAO value of mice in group C was the lowest, which indicates that the diet of group C can promote the growth of beneficial bacteria, balance microbial homeostasis, and inhibit microorganisms that can produce trimethylamine oxide precursors.

[0127] Short-chain fatty acids (SCFAs) play a vital role in maintaining human gut health. Butyrate, in particular, acts as a growth inducer and inflammation inhibitor of the colonic mucosa, inducing apoptosis in cancer cells and preventing colon cancer. The level of SCFAs in the gut reflects bacterial activity and influences the regulation of liver lipids and carbohydrates. Bacteria utilize oligosaccharides to produce short-chain fatty acids, which then bind to… Figure 1 The pH values ​​of the intestinal contents of mice in experimental groups A, B, and C were significantly lower than those in the control group. However, analysis of the serum test results in the second week of the experiment revealed that the SCFA level in the serum of mice in the control group was the highest. At the end of the third week of the experiment, the serum results in group C showed no significant change. This result is related to the reduced permeability of the thickened intestinal mucosa, which is consistent with the lowest DAO level in the serum of group C.

[0128] In summary, the three experimental groups (A, B, and C) represent three types of formulations. Group C is the most ideal formulation; the synergistic effect of its components positively impacted gut microbiota homeostasis and the host without producing side effects. Groups A and B, while also positively impacting gut microbiota homeostasis and the host, both caused some side effects at certain stages. This was further confirmed in subsequent experimental results.

[0129] 3.3 Effects on the height of small intestinal villi and crypts

[0130] Table 2. Changes in intestinal villus height and crypt depth after dietary intervention in the experimental group.

[0131]

[0132] From Table 2 and Appendix Figure 5 It can be seen that the indicators of experimental groups A, B, and C differed from those of the blank group O mice after 14 and 21 days of feeding. The small intestine is the main site for the digestion, absorption, and transport of nutrients in the body; therefore, a good small intestinal mucosal structure is particularly important for perfecting digestive physiological functions and promoting the growth and development of the body. As an important component of the small intestine, the intestinal villi not only play a crucial role in the absorption of nutrients, but also suffer from the strong vibrations that disrupt the colonization of harmful bacteria. The effects of probiotics on intestinal morphology and function may extend beyond changes in villus morphology to include more significant changes at the cellular level. The ratio of villus height to crypt depth reflects the functional state of the small intestine. A decrease in this ratio indicates mucosal damage, decreased digestive and absorptive functions, and impaired growth and development. After 14 days of feeding, the ratio of villus height to crypt depth in groups A and B was higher than that in the control group, while the difference in group C was not significant. After 21 days, the ratio of villus height to crypt depth in groups B and C was higher than that in the control group, with group C showing the best performance. This may be because it promotes the establishment of a normal gut microbiota in mice, maintains the stability of the digestive tract environment, and improves the environment for the presence of intestinal villi. This was also verified in the blood biochemical test of diamine oxidase (DAO) values ​​in group C.

[0133] 3.4 Measurement of intestinal contents metabolomics

[0134] Table 3. Changes in the values ​​of some organic acids in the experimental group

[0135]

[0136] Table 3 shows the changes in the content of isoleucine, leucine, valine, tyrosine, succinic acid, oxalic acid, valerate, and isovaleric acid in the intestinal contents of mice fed different diets. Diabetes mellitus leads to abnormal metabolism of some amino acids, with two main characteristics: 1. The total plasma amino acid content and glucogenic amino acid content decrease in diabetes, and are significantly negatively correlated with blood glucose; 2. Regardless of the level of blood glucose control, the content of branched-chain amino acids and their proportion of total amino acids both increase.

[0137] However, 5-10 years before the onset of diabetes, the levels of some amino acids are elevated, especially five amino acids in the blood: isoleucine, leucine, valine, tyrosine, and phenylalanine. This experiment clearly shows, particularly the results after 21 days, that these five amino acids changed significantly with a rational feed formulation, primarily showing a marked decrease. This has significant implications for the prevention and reduction of diabetes incidence.

[0138] Assessing these amino acid indicators is not only significant for predicting the risk of developing diabetes, but also for heart disease and tumors (kidney cancer, pancreatic cancer), enabling targeted early intervention and prevention, and playing a precise primary prevention role.

[0139] Bacteria utilize oligosaccharides to produce short-chain fatty acids, and their increased levels lead to a decrease in intestinal pH. Acetic acid is mainly produced by Bacteroides, while butyrate is primarily produced by Firmicutes. Butyrate is a major energy source for colonic cells, increasing intestinal health and potentially reducing intestinal permeability and preventing metabolic endotoxemia. At 14 days of feeding, compared with A and B mice, group C showed increased levels of succinic acid and oxalic acid, and decreased levels of valeric acid; at 21 days of feeding, oxalic acid levels increased, while succinic acid and isovaleric acid levels decreased.

[0140] 3.5 16SDNA assay of intestinal contents

[0141] In this experiment, bacteria from four phyla were detected in mouse intestinal contents samples, with Bacteroidetes, Firmicutes, and Proteobacteria being the three most abundant phyla. Figures 6 to 9 The phylum-level composition of the gut microbiota in mice after 21 days is shown. It can be seen that in the control group (0 mice), the abundance of Bacteroidetes reached 65.6%, Firmicutes 31.66%, and Proteobacteria 1.44%.

[0142] In experimental group A mice, the abundance of the three reached 87.84%, 2.98%, and 8.77%, respectively;

[0143] In experimental group B mice, the abundance of the three reached 90.72%, 5.93%, and 2.75%, respectively;

[0144] In experimental group C mice, the abundance of the three reached 85%, 65%, 7.00%, and 6.45%, respectively.

[0145] Compared with the control group, experimental groups A, B, and C showed an increase in the abundance of Bacteroidetes and Proteobacteria, and a decrease in the abundance of Firmicutes. This indicates that the ingestion of oligosaccharides by mice promoted the proliferation of Bacteroidetes and Proteobacteria, while inhibiting the growth of Firmicutes.

[0146] In this experiment, bacteria from 14 genera were detected in mouse intestinal contents, with proportions exceeding 0.1%. *Bacteriodes*, *Alloprevotella*, *Parabacteriodes*, *Lachnospiraceae*, and *Parasutterella* were the predominant genera in the intestinal contents samples. Figure 10 The genus-level composition of the gut microbiota in mice after 21 days shows that:

[0147] After mice were fed diet A, the amount of Bacteriodes in the intestinal contents increased significantly, while the amount of Bacteriodes in the intestinal contents of mice in group B decreased significantly, and the change in Bacteriodes in group C was not obvious. This indicates that the functional oligosaccharide composition has the effect of promoting the proliferation of Bacteriodes, while ordinary oligosaccharides do not have this effect.

[0148] The levels of Alloprevotella and Parabacteriodes increased in the intestinal contents samples of mice in groups B and C, while they decreased in group A, indicating that common oligosaccharides have the ability to promote the proliferation of Alloprevotella and Parabacteriodes.

[0149] Compared to the control group, Lachnospiraceae levels decreased in groups A, B, and C, while Parasutterella levels increased. Galacto-oligosaccharides and inulin, as prebiotics, are not digested and absorbed by the body but directly enter the large intestine, playing a crucial role in adjusting gut microbiota, maintaining a normal gut environment, regulating intestinal function, and improving overall health. Polydextrose, a water-soluble dietary fiber, can shorten gastric emptying time, effectively improving intestinal function. After ingestion, it is fermented only in the lower part of the gastrointestinal tract, producing short-chain fatty acids such as butyric acid, which lowers intestinal pH and helps resist infection. Insoluble dietary fiber cannot dissolve in water or be fermented by microorganisms in the large intestine. However, it can balance the metabolism of protein, fat, and carbohydrates, slow down the release of nutrients, promote balanced development of small intestinal tissue, increase overall intestinal motility, reduce the residence time of excrement in the intestine, and increase the volume of intestinal contents, thus playing an important role in improving the microbial environment and promoting bowel movements.

[0150] This experiment focuses on verifying the synergistic effect of functional polysaccharides and ordinary polysaccharide compositions. This synergistic effect focuses on two aspects: first, balancing the effects of anti-inflammatory nutrition on changes in host physiological indicators.

[0151] Second, the effects of balancing anti-inflammatory nutrition on the microbial environment were investigated. Experimental results showed that all types of polysaccharide compositions had significant effects on the physiological indicators and intestinal microecology of mice. Among them, the synergistic effect of functional polysaccharide and ordinary polysaccharide compositions produced positive effects on the host without producing side effects. The experimental groups that added functional polysaccharide compositions or ordinary polysaccharide compositions alone produced positive effects on the host, but also caused certain side effects in stages.

[0152] 3.6 Experimental Conclusions and Analysis:

[0153] This experiment investigated the effects of different polysaccharide compositions on mice. Four different feed formulations were used to feed the mice: group O, group A, group B, and group C. Genomic analysis of the intestinal microbiota revealed that different diets significantly impacted the gut microbiota. Furthermore, analysis of colonic contents pH and serum SCFA data showed that the fermentation of oligosaccharides and dietary fiber produced large amounts of short-chain fatty acids, leading to a decrease in pH. Analysis of serum DAO and TMAO levels and intestinal villus data revealed that group C mice performed best, indicating that for normal mice, a diversity of polysaccharides and dietary fiber contributes to a healthy gut system.

[0154] In summary, all types of polysaccharide compositions significantly affected the physiological indicators and gut microbiota of mice. The synergistic effect of compositions containing functional and ordinary polysaccharides produced positive effects on the host without causing side effects. In contrast, experimental groups supplemented with either functional or ordinary polysaccharide compositions alone, while producing positive effects on the host, also experienced some side effects at certain stages. This indicates that nutritional fortifiers and supplements such as ordinary polysaccharides are important nutritional components and key elements for maintaining health.

[0155] Application Example 2

[0156] This embodiment selected 40 volunteer subjects for human trials, and examined changes in various indicators before and after 45 days of the trial. The specific human trial steps are as follows:

[0157] 1. Target audience selection

[0158] Inclusion criteria:

[0159] 1.1 Age: 18-75 years old;

[0160] 1.2 Informed consent, volunteer participants.

[0161] 1.3 Anyone who meets the above two criteria can be included in the trial cases.

[0162] Exclusion criteria:

[0163] 1.4 Age > 75 years or < 18 years.

[0164] 1.5 Patients with major cardiovascular and cerebrovascular diseases, such as stroke, severe hypertension, heart failure, major infectious diseases, and mental illness.

[0165] 1.6 Individuals who have participated in other clinical trials within the past 3 months.

[0166] 2. Tasting Program

[0167] 2.1 Trial Food: The anti-inflammatory combination food was provided by Beijing Ruiqianjing Technology Development Co., Ltd., and all raw materials used were food and food ingredients. The food included inulin, galactooligosaccharides, polydextrose, and water-insoluble dietary fiber, with a weight ratio of 25:25:20:30.

[0168] 2.2 Trial method: Three packets per day, 10 grams per packet, taken with warm water before meals (immediately after eating).

[0169] 2.3 Trial period: 45 consecutive days of use.

[0170] 2.4 Precautions: The test group should take the test food according to the recommended method and dosage. During the trial, maintain normal dietary habits and medication habits. If constipation improves during the trial, discontinue laxatives as appropriate. Avoid excessive alcohol consumption during the trial.

[0171] 3. Testing Time Points and Testing Content

[0172] Table 4. Detection Time Points and Detection Contents of the Human Food Trial

[0173]

[0174] 4. Testing Items

[0175] 4.1 Basic Information: Blood Pressure, Weight, Constipation

[0176] 4.2 Complete blood count (complete blood count and liver function tests in one tube)

[0177] 4.3 Sampling

[0178] 4.4 Liver function and fasting blood glucose (diabetic patients should self-test their blood glucose two hours after a meal and record it on the CRF form and investigator's manual)

[0179] 4.5 Blood lipids (triglycerides)

[0180] 4.6 Improvement of cellular metabolic capacity: amino acids (leucine, isoleucine, phenylalanine, tyrosine, valine).

[0181] 4.7 Intestinal function tests: DAO, D-lactate, TMAO

[0182] 4.8 Serum inflammatory factors

[0183] 5. Testing methods and instruments used

[0184] 5.1 Complete blood count: Blood routine analyzer BC3000 (testing unit: Beijing Hyster Medical Laboratory Co., Ltd.).

[0185] 5.2 Liver function: Roche cobas c501 chemiluminescence assay (testing unit: Beijing Hyster Medical Laboratory Co., Ltd.).

[0186] 5.3 Blood glucose and blood lipids: Blood glucose (Roche cobas c501), blood lipids (Roche cobas c701).

[0187] 5.4 Amino acid detection: High performance liquid chromatography-quadrupole ion trap tandem mass spectrometry API3200Q-TRAP (testing unit: Beijing Mass Spectrometry Medical Research Co., Ltd.).

[0188] 5.5 Serum DAO and serum D-lactic acid: (Testing unit: Beijing Mass Spectrometry Medical Research Co., Ltd.)

[0189] 5.6 Serum TMAO: Mass spectrometry detection by LC-MS / MS (Agilent 6430) (Testing unit: Academy of Military Medical Sciences).

[0190] 5.7 Serum inflammatory factors: protein liquid phase suspension chip (Luminex 200) (testing unit: Academy of Military Medical Sciences).

[0191] 6. Test Results

[0192] 6.1 Results of Differences in Blood Biochemical Indicators

[0193] Differences in blood biochemical indicators were compared among all 40 individuals (the general population), 10 individuals with high BMI and diabetes, and 25 healthy individuals (excluding those with lung cancer, psoriasis, etc.). The results are as follows: Figure 11-13 See Table 5.

[0194] Table 5. Results of differences in blood biochemical indicators among the participants after 45 days of trial.

[0195]

[0196] Note: In the table, "↓" indicates that the trend of each indicator after the food tasting is downward compared to before the food tasting; "↑" indicates that the trend of each indicator after the food tasting is upward compared to before the food tasting; "---" indicates that there is no significant change in each indicator after the food tasting compared to before the food tasting.

[0197] From the appendix Figure 11-13 According to the specific numerical analysis and the summary statistics in Table 5 above, (1) the blood insulin levels of all participants in this experiment increased, as shown in the attached table. Figure 11 As shown in E, pancreatic function is improved. Combined with hematological, immunological, and diabetes-related amino acid metabolism results, this anti-inflammatory composition has significant effects on the prevention of diabetes.

[0198] (2) The results of this study showed that uric acid levels were significantly reduced in all participants, as shown in the attached figure. Figure 12 As shown in Figure A, nucleic acid metabolism is improved. Combined with the results of hematological and immunological comprehensive indicators, it is shown that this anti-inflammatory composition has significant effects on improving nucleic acid metabolism, preventing gout, and improving renal function in response to the current situation of high uric acid levels.

[0199] (3) The results of this study showed that blood lead levels were significantly reduced in all participants, as shown in the attached figure. Figure 12 As shown in Figure B, this anti-inflammatory composition has significant implications for improving blood environment and reducing heavy metal damage in the context of today's heavily polluted environment.

[0200] (4) The results of this study showed that all participants experienced a decrease in alanine aminotransferase (ALT), indirect bilirubin, and total bilirubin, as shown in the attached figure. Figure 12 As shown in F, 13B, and 13A, liver function was improved. Combined with the results of hematological and immunological comprehensive indicators, it is shown that this anti-inflammatory composition has significant effects on improving liver function.

[0201] (5) The results of this study showed that all participants improved their hemoglobin, mean corpuscular hemoglobin (MCH) content, mean corpuscular hemoglobin concentration, and globulin hematological parameters, as shown in the attached figure. Figure 11 As shown in A, 11B, 11C, and 12D, the results, combined with immunological and nutritional metabolic findings, indicate that this anti-inflammatory composition has significant implications for improving sub-health conditions.

[0202] (6) The results of this study showed that all participants experienced a reduction in low-density cholesterol and triglycerides, as shown in the attached figure. Figure 13 As shown in D and 11F, blood insulin levels increased, as indicated in the appendix. Figure 11 As shown in E, this demonstrates improved pancreatic function. Combined with the results of hematological and immunological comprehensive indicators, this is of great significance for improving lipid metabolism and preventing cardiovascular and cerebrovascular diseases.

[0203] (7) The results of this study showed that albumin, albumin-to-globulin ratio, and immunoglobulin IgA were increased in all participants, as shown in the attached figure. Figure 12 As shown in C, 12E, and 11D, enhancing immunity, combined with the results of immunology and nutritional metabolism, improves the basic elements of human health.

[0204] 6.2 Results of differences in amino acid indicators

[0205] The amino acid levels of all 40 participants (the entire trial population), 10 individuals with high BMI and diabetes, and 25 healthy individuals (excluding those with lung cancer, psoriasis, etc.) were compared. The results are as follows: Figure 14-15 See Table 6.

[0206] Table 6. Results of differences in amino acid levels among the population after 45 days of trial feeding.

[0207]

[0208] Note: In the table, "↓" indicates that the trend of each indicator after the food trial is downward compared to before the food trial; "↑" indicates that the trend of each indicator after the food trial is upward compared to before the food trial; "---" indicates that there is no significant change in each indicator after the food trial compared to before the food trial.

[0209] Nutritional metabolism is the foundation of health. Amino acid (protein) metabolism is closely related to lipid metabolism and glucose metabolism. Diabetes-related abnormal amino acid metabolism has two main characteristics: 1. The total amino acid content and glucogenic amino acid content in plasma decrease in diabetes, and are significantly negatively correlated with blood glucose; 2. Regardless of the quality of blood glucose control, the content of branched-chain amino acids and their proportion of total amino acids increase.

[0210] However, 5-10 years before the onset of diabetes, the levels of some amino acids are elevated, especially five amino acids in the blood: isoleucine, leucine, valine, tyrosine, and phenylalanine. The results of this experiment show that after consuming the anti-inflammatory composition of this application for 45 days, the sum of these five amino acids significantly decreased, as shown in the attached figure. Figure 14 and 15 As shown, this has significant implications for preventing the incidence of diabetes.

[0211] 6.3 Results of differences in intestinal permeability indicators

[0212] Differences in intestinal permeability in all 40 individuals (all participants), 10 individuals with high BMI and diabetes, and 25 healthy individuals (excluding those with lung cancer, psoriasis, etc.) were compared. The results are as follows: Figure 16 See Table 7.

[0213] Table 7. Results of differences in intestinal permeability indicators among the population after 45 days of trial feeding.

[0214]

[0215] Note: In the table, "↓" indicates that the trend of each indicator after the food trial is downward compared to before the food trial; "↑" indicates that the trend of each indicator after the food trial is upward compared to before the food trial; "---" indicates that there is no significant change in each indicator after the food trial compared to before the food trial.

[0216] This set of indicators are landmark indicators for measuring intestinal permeability. Trimethylamine oxide (TMAO) is an exceptionally sensitive indicator; in animal studies, most functional polysaccharides, when used alone, tend to worsen this indicator. However, this study showed no difference in the entire trial population, demonstrating that the synergistic effect of the formulation compensates for the deficiencies of the functional polysaccharide composition. D-lactic acid is a rigid indicator representing the level of intestinal permeability; its significant decrease is shown in the attached figure. Figure 16 As shown in Figure A, this indicates a decrease in the entry of the intestinal metabolite D-lactic acid into the bloodstream, resulting in a significant improvement in intestinal function. Glutamine levels are also reduced, as shown in the attached figure. Figure 16 As shown in Figure B, this demonstrates improved cell utilization, particularly of intestinal mucosal epithelial cells. This aligns with the improved bowel function, including constipation, observed in all participants in the trial. Reduced intestinal throughput and improved bowel function are crucial foundations for overall health improvement.

[0217] 6.4 Results of Cytokine Differences

[0218] The cytokine levels of all 40 participants (the entire trial population), 10 individuals with high BMI and diabetes, and 25 healthy individuals (excluding those with lung cancer, psoriasis, etc.) were compared. The results are as follows: Figure 17-20 See Table 8.

[0219] Table 8. Results of differences in cytokine levels among the population after 45 days of trial.

[0220]

[0221] Note: In the table, "↓" indicates that the trend of each indicator after the food trial is downward compared to before the food trial; "↑" indicates that the trend of each indicator after the food trial is upward compared to before the food trial; "---" indicates that there is no significant change in each indicator after the food trial compared to before the food trial.

[0222] Immune balance is crucial for maintaining human health. Modern populations, due to external pressures and unhealthy lifestyles, are often in a "sub-healthy state," where their immune systems are frequently subjected to various stresses or malfunction, leading to inflammatory responses. This inflammatory response lies between a basal homeostatic state and a classic inflammatory response, and is termed low-grade chronic inflammation or para-inflammation. Cytokines are a class of small-molecule proteins with broad biological activity that are synthesized and secreted by immune cells or certain non-immune cells in response to external stimuli. They mainly include interleukins (IL), tumor necrosis factor (TNF), interferons (IFN), chemokines, and colony-stimulating factors (CSF), and can be used as biomarkers for the diagnosis of inflammation or diseases.

[0223] The five inflammatory factors that decreased in this trial—IL-12p70, IL-13, IL-2, IL-23, and RANTES—are listed below. Figure 17As shown, IL-12 and IL-23 are mainly produced by antigen-presenting cells such as dendritic cells (DCs) and macrophages. IL-12 can promote the differentiation of inflammatory Th1 cells, while IL-23 can maintain the activity of inflammatory Th17 cells and inhibit the activity of anti-inflammatory Treg cells. IL-2 is mainly produced by inflammatory Th1 cells. IL-13 is mainly produced by anti-inflammatory Th2 cells. RANTES is significantly upregulated after T cell activation and can chemotactically attract activated T cells. This anti-inflammatory composition can significantly inhibit four pro-inflammatory factors, namely IL-12, IL-2, IL-23, and RANTES, indicating that it may improve sub-inflammatory conditions by inhibiting related inflammatory T cells.

[0224] As attached Figure 18-20 As shown, among the cytokines that increased in this experiment, IL-1RA is a natural IL-1 receptor antagonist and can inhibit the inflammatory response of the pro-inflammatory IL-1 pathway. Studies have shown that IL-1RA is significantly decreased in the intestinal mucosa of CD and UC patients, predicting the activation of the pro-inflammatory IL-1 pathway in disease states. The results of this study indicate that the combined treatment significantly increased the in vivo IL-1RA secretion level (from 0 before intervention to an average of 314 pg / ml after intervention). IL-4 can promote the differentiation and maturation of anti-inflammatory Th2 cells, and IL-27 can induce T cells to produce IL-10 under certain conditions, thus exerting an immunosuppressive effect. The combined treatment significantly increased the in vivo secretion levels of IL-4 and IL-27. Of course, not all cytokines are ideal; this is related to the duration of use, cell metabolic cycle, and bodily regulation. Further measurements are needed to assess the improvement in cytokine regulation with prolonged use.

[0225] 6.5 Results of Gut Microbiota Differences

[0226] Before and after the food trial in the entire population, the Wilcoxon test was used to examine the differences in relative abundance of bacterial communities at the phylum, class, order, family, genus and species levels between paired samples, and the comparison results were corrected for P values ​​by FDR (p.adj<0.05). The differences at the species level are shown in Table 9 below.

[0227] Table 9. Results of comparisons among bacterial groups

[0228]

[0229] As shown in Table 9 above, there were significant differences in the gut microbiota among all participants, which is consistent with the results of biochemical and intestinal function tests.

[0230] This invention presents an anti-inflammatory composition that improves nutrient metabolism, intestinal function, and intestinal microecology. Through a rational combination of water-insoluble dietary fiber and functional polysaccharides, it adjusts the release rate of nutrients in the intestine, slows digestion, accelerates cholesterol excretion, absorbs and eliminates toxic substances from food, and particularly improves the metabolism of branched-chain amino acids and aromatic amino acids related to diabetes prevention. It also ensures the stability or reduction of blood trimethylamine oxidase biochemical indicators, reduces intestinal permeability, and simultaneously possesses water-absorbing and swelling, gradient adhesion, mechanical isolation, mesh adsorption, ion exchange, and microbial regulation functions, providing a favorable environment and food for intestinal microbial growth and maintaining intestinal microbial homeostasis. The synergistic effect of the anti-inflammatory composition eliminates the negative effects of using it alone, reduces blood endotoxins, blood lead, and inflammatory factors, enhances cellular metabolism and immune capacity, eliminates pathogenic factors such as chronic inflammation and low immunity, and effectively prevents the occurrence of chronic diseases such as cardiovascular and cerebrovascular diseases and diabetes. This invention's anti-inflammatory composition effectively improves nutrient metabolism, reduces intestinal permeability, maintains the homeostatic environment of intestinal microorganisms, and prevents chronic diseases.

[0231] The anti-inflammatory food containing the above-mentioned anti-inflammatory composition is an indispensable staple food for people every day. It provides the human body with a balanced nutritional composition, improves physical condition, and effectively prevents foodborne chronic diseases, thus ensuring health.

[0232] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any simple modifications, equivalent changes, or alterations made by those skilled in the art using the disclosed technical content shall fall within the protection scope of the present invention.

Claims

1. The use of a composition in the preparation of food or medicine that reduces blood lead levels, characterized in that, The composition is a composition for improving intestinal microecology, comprising inulin, galactooligosaccharides, polydextrose and water-insoluble dietary fiber, wherein the weight ratio of inulin, galactooligosaccharides, polydextrose and water-insoluble dietary fiber is 25:25:20:30, and the composition is taken with warm water before meals and immediately after eating.

2. The use of the composition according to claim 1 in the preparation of food or medicine that reduces blood lead levels, characterized in that, The inulin is fructooligosaccharide, fructooligosaccharide, or a mixture of fructooligosaccharide and fructooligosaccharide; the water-insoluble dietary fiber is cellulose, hemicellulose, lignin, or a mixture thereof that is insoluble in water.

3. The use of the composition according to claim 2 in the preparation of food or medicine that reduces blood lead levels, characterized in that, The water-insoluble dietary fiber is wheat bran fiber extracted from wheat bran.

Images