Children formula food for improving intestinal inflammation and / or microbial disorder as well as preparation method and application of children formula food

Abstract

The invention provides a children formula food for improving intestinal inflammation and / or microbial disorder as well as a preparation method and application of the children formula food. The invention firstly provides the infant formula food, the infant formula food is infant formula food or child formula food, the infant formula food comprises 2 '-fucosyllactose and lactose-N-neotetraose, and the mass ratio of the 2'-fucosyllactose to the lactose-N-neotetraose is (1-5.5): 1. The children formula food can improve intestinal inflammation and / or microbial disorder.

Description

Technical Field

[0001] The present invention belongs to the technical field of pediatric formula food, and specifically relates to a pediatric formula food containing 2'-fucosyllactose and lactose-N-neotetraose and helping to improve intestinal inflammation and / or microbial imbalance, and a preparation method and application thereof. Background Art

[0002] The first 1000 days of life is a window of opportunity for infant development. The dynamic development of the intestinal microecology complements the development of the intestinal immune system. The intestine is not only an important place for the digestion and absorption of nutrients, but also an important barrier to defend against external pathogens. Maintaining intestinal homeostasis and enhancing intestinal immune function are essential for maintaining body health. After birth, infants and young children are at high risk of gastrointestinal infections, inflammations and allergic diseases due to their immature digestive and immune functions and the unstable formation and development of intestinal flora. Breast milk is the first food that infants come into contact with. Breast milk is not only rich in nutrients, but also rich in immune active ingredients, growth factors and oligosaccharides. These ingredients in breast milk can not only provide immune support for infants and promote the maturation of intestinal barrier function, but also provide nutrients for probiotics such as bifidobacteria and lactobacilli, which helps to form a good microecological environment and ensure intestinal health. Literature results show that the infant's intestinal flora is closely related to the infant's development and the formation of the immune system. Establishing a healthy intestinal flora in the first 1000 days of life is important for the development and maturation of the intestinal mucosal immune system and the systemic immune system. The structure and relative abundance of the intestinal flora in early life will affect the probability of disease in the future.

[0003] In addition, there are a large number of keystone species in healthy humans, which play an important role in the species diversity and functional diversity of intestinal microorganisms and are indispensable. The specific composition of keystone microorganisms in infants and young children still needs to be further clarified. Some literature points out that the intestinal microbiome of infants is mainly composed of five phyla: Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria and Verrucomicrobia. A large number of studies have shown that the difference between the intestinal flora of diseased organisms and healthy organisms is mainly manifested in the decrease in the relative abundance of Firmicutes in the intestines of diseased organisms and the increase in the relative abundance of Proteobacteria.

[0004] Providing a nutritional composition that can prevent and / or improve intestinal inflammatory symptoms and / or microbial imbalance to infants or children is crucial for the healthy growth of infants and children. Summary of the invention

[0005] An object of the present invention is to provide a pediatric formula food that can improve intestinal inflammation and / or microbial imbalance.

[0006] Another object of the present invention is to provide a method for preparing the pediatric formula food.

[0007] Another object of the present invention is to provide the related applications of the pediatric formula food.

[0008] On the one hand, the present invention provides a pediatric formula food, which is an infant formula food or a children's formula food. The pediatric formula food includes 2'-fucosyllactose and lacto-N-neotetraose, and the mass ratio of 2'-fucosyllactose to lacto-N-neotetraose is (1 - 5.5):1.

[0009] 2'-fucosyllactose (2'-FL or 2-FL) is a trisaccharide structure formed by fucose and lactose, and is a representative substance of fucosylated oligosaccharides. Commercially available 2'-FL is usually prepared by microbial fermentation and has the same structure as the oligosaccharides found in human milk.

[0010] Lacto-N-neotetraose (LNnT) is an oligosaccharide composed of galactose, glucose, and N-acetylglucosamine, and is a representative substance of neutral oligosaccharides in breast milk. Commercially available LNnT is usually prepared by microbial fermentation and has the same structure as the oligosaccharides found in human milk.

[0011] The present invention finds through research that combining 2'-fucosyllactose and lacto-N-neotetraose in a specific ratio has a synergistic effect on improving intestinal inflammation symptoms, improving intestinal microbial dysbiosis, and restoring key intestinal flora.

[0012] In the present invention, unless otherwise specified, the pediatric includes infants, toddlers, or children. The infant is an infant aged 0 - 12 months (including infants aged 0 - 6 months and older infants aged 6 - 12 months), the toddler refers to a toddler aged 1 - 3 years, and the child refers to a child aged 3 years to 18 years (under 19 years old), especially a younger child aged 3 years to 6 years (under 7 years old).

[0013] According to specific embodiments of the present invention, the pediatric formula food of the present invention is infant formula food (infant formula, toddler formula) or children's formula food (such as children's modified milk powder). Among them, in the dried matter equivalent of the infant formula food, the total protein content is 7.8 g to 22.8 g / 100 g (preferably, the proportion of whey protein in the total protein is 40% to 75%), the fat content is 14.9 g to 33.8 g / 100 g, and the carbohydrate content is 40.7 g to 79.3 g / 100 g. Among them, in the dried matter equivalent of the children's formula food, the total protein content is 16.5 g to 30 g / 100 g, the fat content is 10 g to 35 g / 100 g; the carbohydrate content is 40 g to 70 g / 100 g.

[0014] According to some specific embodiments of the present invention, the pediatric formula food of the present invention is infant formula for special medical purposes or toddler formula for special medical purposes.

[0015] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the mass ratio of 2'-fucosyllactose to lacto-N-neotetraose is (1 - 4.8):1.

[0016] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the mass ratio of 2'-fucosyllactose to lacto-N-neotetraose is (1 - 4):1.

[0017] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the mass ratio of 2'-fucosyllactose to lacto-N-neotetraose is 2:1.

[0018] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the content of 2'-fucosyllactose is 0.5 - 1.8 g / 100 g of food dry matter. The amount of lacto-N-neotetraose can be determined with reference to the aforementioned ratio range of 2'-fucosyllactose to lacto-N-neotetraose.

[0019] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the content of 2'-fucosyllactose is 0.6 - 1.6 g / 100 g of food dry matter.

[0020] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the content of 2'-fucosyllactose is 0.7 - 1.4 g / 100 g of food dry matter.

[0021] According to some specific embodiments of the present invention, in the pediatric formula food of the present invention, the content of 2'-fucosyllactose is 0.8 - 1.3 g / 100 g of food dry matter.

[0022] According to some specific embodiments of the present invention, the amount of 2'-fucosyllactose in the infant formula food of the present invention is 0.6 - 0.7 g / 100 g of food dry matter, 0.8 - 0.9 g / 100 g of food dry matter, 1.0 - 1.1 g / 100 g of food dry matter, 1.2 - 1.3 g / 100 g of food dry matter, 1.4 - 1.5 g / 100 g of food dry matter or 1.6 - 1.7 g / 100 g of food dry matter.

[0023] According to a specific embodiment of the present invention, in the infant formula food of the present invention, the raw materials providing total protein include one or more of raw milk (cow / sheep), whole milk powder (cow / sheep), skim milk powder (cow / sheep), whey protein powder (cow / sheep), demineralized whey powder (cow / sheep), β-casein, etc.

[0024] According to a specific embodiment of the present invention, in the infant formula food of the present invention, in addition to the basic raw materials containing milk fat, the raw materials providing fat may also include vegetable oil or OPO structured lipid.

[0025] According to a specific embodiment of the present invention, for the infant formula food of the present invention, a part of the carbohydrates can come from basic raw materials containing lactose such as milk, whole milk powder and / or skim milk powder, etc. In addition, lactose raw materials can be additionally added to provide carbohydrates.

[0026] According to a specific embodiment of the present invention, in the infant formula food of the present invention, in addition to the above components, it may also include conventional components of infant formula food or children's modified milk powder, for example, it may also include one or more of nutrients, dietary fiber, inositol, L-carnitine, docosahexaenoic acid, arachidonic acid, lutein, nucleotides.

[0027] In a specific embodiment, the infant formula food of the present invention is a dairy product, which can be a solid dairy product or a liquid dairy product. Specifically, it can be in the form of cheese, milk tablets, milk powder or liquid milk.

[0028] It can be understood that for different types of infant formula foods, the content of each functional substance in the final product should be appropriately adjusted within the range permitted by relevant standards and specifications.

[0029] In some specific embodiments of the present invention, the pediatric formula food of the present invention is milk powder, which can be prepared by adding / adjusting 2'-fucosyllactose and lacto-N-neotetraose to a basic formula powder. Among them, after the milk powder of the present invention is prepared, the contents of 2'-fucosyllactose and lacto-N-neotetraose can be determined by using conventional methods in the art, such as high performance liquid chromatography detection and analysis techniques. There is no particular limitation on the basic formula powder in the present invention, as long as it is a commonly used milk powder formula in the art. For example, the basic formula powder can be a common infant formula powder, toddler formula powder or children's milk powder. As a specific example, the basic formula powder of the present invention can take milk powder of animal origin as the main raw material, and can further be added with one or more of whey powder, vitamins, minerals, etc. As the milk powder of animal origin, cow milk powder or goat milk powder can be used.

[0030] According to some specific embodiments of the present invention, the pediatric formula food of the present invention has the following raw material composition:

[0031] Raw cow milk (partially skimmed available): 700 - 3700 parts (for example: 800 parts, 1000 parts);

[0032] Whey powder: 0 - 600 parts (which can be converted into the corresponding amount of whey liquid, for example: 300 parts, 200 parts);

[0033] Whey protein powder and / or lacto-N-neotetraose raw material: 0 - 125 parts; preferably including lacto-N-neotetraose raw material: 1.5 - 22 parts; for example, whey protein powder: 0 - 100 parts (which can be converted into the corresponding amount of whey protein liquid, for example: 32 parts, 25 parts); raw material dedicated to providing lacto-N-neotetraose with higher purity: 1.5 - 22 parts (for example: 8 parts, 10 parts);

[0034] Edible vegetable blended oil (may contain 1,3-dioleoyl-2-palmitoyl glycerol): 80 - 240 parts (for example: 230 parts, 180 parts);

[0035] Lactose: 0 - 550 parts (for example: 200 parts, 245 parts);

[0036] Minerals: 8 - 30 parts (for example: 18 parts, 15 parts);

[0037] Trace elements: 0.5 - 2 parts (for example: 1 part, 1.5 parts);

[0038] Vitamins: 2.5 - 10 parts (for example: 5 parts, 6 parts);

[0039] α-Lactalbumin (providing high-quality protein): 0.625 - 3 parts (for example: 1.75 parts, 1.5 parts);

[0040] Linoleic acid (providing essential fatty acids): 2.5 - 5 parts (e.g., 5 parts, 4 parts);

[0041] 2'-Fucosyllactose: 2.5 - 13 parts (e.g., 5 parts, 10 parts);

[0042] Lacto-N-neotetraose: 2.5 - 5 parts (e.g., 3 parts, 4 parts).

[0043] According to a specific embodiment of the present invention, the raw material composition of the pediatric formula food of the present invention may further include one or more of prebiotics (such as galactooligosaccharides, fructooligosaccharides, etc., providing support for intestinal health), probiotics, and the like.

[0044] According to a specific embodiment of the present invention, in the pediatric formula food of the present invention, each component raw material can be commercially available.

[0045] In the food of the present invention, the amounts of various substances should meet the requirements of relevant standards.

[0046] On the other hand, the present invention also provides a method for preparing the pediatric formula food described above, and the method includes:

[0047] Using a wet method, a dry method, or a wet-dry composite production process, the raw materials providing 2'-fucosyllactose and lacto-N-neotetraose are mixed with other raw materials in the formula. After the mixed material is filtered to remove impurities, it is homogenized and further used to prepare the pediatric formula food.

[0048] According to some specific embodiments of the present invention, in the method for preparing the pediatric formula food of the present invention, the homogenization temperature is not lower than 50 °C and the pressure is not lower than 120 bar.

[0049] According to some specific embodiments of the present invention, the method for preparing the pediatric formula food of the present invention further includes:

[0050] The homogenized material is concentrated until the dry matter weight content of the concentrate is 40% - 60%;

[0051] The concentrated material is sterilized at a temperature above 80 °C for 10 - 60 seconds;

[0052] The sterilized material is dried through a spray drying tower (inlet air temperature 150 °C - 200 °C, outlet air temperature 80 °C - 120 °C, negative pressure -10 - -2 mmWG, high-pressure pump pressure 140 - 250 bar), and then introduced into a fluidized bed for secondary drying to obtain a powder.

[0053] In the preparation method of the present invention, ① according to the addition of 2'-fucosyllactose and lacto-N-neotetraose, the formula is optimized, which can improve intestinal inflammation and / or microbial dysbiosis, and ensure that infants and children can fully absorb and utilize these key nutrients. ② Mixing and formulation: The components are uniformly mixed in a specific ratio to ensure the consistency of the nutritional components in each formula food and avoid nutritional imbalance. Uniform mixing helps 2'-fucosyllactose and lacto-N-neotetraose play their best roles in the intestine and ensure the consistency of the product effects. ③ Production and processing: Advanced production processes such as homogenization and sterilization are adopted to ensure the safety and quality of the product, extend the shelf life, and at the same time retain the activity and stability of the nutritional components.

[0054] On the other hand, the present invention also provides the use of the pediatric formula food described above in the preparation of a product for improving intestinal inflammation symptoms and / or microbial dysbiosis. The product can be the pediatric formula food of any of the foregoing solutions of the present invention, or can be other products further prepared based on the pediatric formula food of the present invention.

[0055] According to a specific embodiment of the present invention, the improvement of intestinal inflammation symptoms and / or microbial dysbiosis includes one or more of the following:

[0056] Improving weight loss caused by intestinal inflammatory response;

[0057] Improving intestinal injury caused by inflammatory response;

[0058] Reducing the increase in serum myeloperoxidase (MPO) activity caused by intestinal inflammatory response;

[0059] Reducing the increase in serum D-lactic acid level caused by intestinal inflammatory response;

[0060] Reducing the increase in serum diamine oxidase (DAO) level caused by intestinal inflammatory response;

[0061] Improving intestinal barrier function;

[0062] Enhancing intestinal physical and / or mucus barrier;

[0063] Increasing the expression level of intestinal mucin;

[0064] Increasing the expression levels of intestinal occludin, claudin, and / or zonula occludens (ZOs);

[0065] Maintaining intestinal cell immune homeostasis;

[0066] Improving intestinal immune cell dysregulation caused by inflammatory response;

[0067] Enhance the expression of intestinal CD4+ T cells;

[0068] Reduce the expression of intestinal CD8+ T cells;

[0069] Increase the ratio of intestinal CD4+ / CD8+ T cells;

[0070] Enhance the diversity and / or community richness of the intestinal microbiota;

[0071] Restore the intestinal keystone microbiota disorder or reduced diversity caused by inflammation or antibiotics;

[0072] Improve the reduction of intestinal microbial species caused by intestinal mucosal immune damage;

[0073] Increase the abundance of intestinal genera Muribaculaceae and / or Ruminococcus;

[0074] Reduce the abundance of intestinal genera Escherichia-Shigella and / or Alloprevotella.

[0075] Overall, the present invention mixes 2'-fucosyllactose and lacto-N-neotetraose in a certain proportion and uses them to prepare pediatric formula foods, which helps to improve intestinal inflammation and / or microbial dysregulation and is beneficial to the healthy growth of infants and children. Brief Description of the Drawings

[0076] Figure 1 Show the changes in body weight of rats in each group after LPS treatment for 6 h in a specific experiment of the present invention.

[0077] Figure 2 Show the weight loss of rats in each group after LPS treatment for 6 h in a specific experiment of the present invention.

[0078] Figure 3 Show the MPO activity in the sera of rats in each group in a specific experiment of the present invention.

[0079] Figures 4A - 4B Respectively show the levels of D-lactic acid and DAO in the sera of rats in each group in a specific experiment of the present invention.

[0080] Figures 5A - 5D Respectively show the expression of mucin and tight junction proteins in the colon tissues of rats in each group in a specific experiment of the present invention.

[0081] Figures 6A - 6C Respectively show the expression of immune cells in each group in a specific experiment of the present invention.

[0082] Figure 7 Show the effect of different groups on the number of OUTs in a specific experiment of the present invention.

[0083] Figure 8 Show the effects of different groups on the intestinal flora composition in a specific experiment of the present invention. Detailed implementation manners

[0084] Before further describing the specific implementation manners of the present invention, it should be understood that the protection scope of the present invention is not limited to the following specific implementation manners; it should also be understood that the terms used in the embodiments of the present invention are for describing specific implementation manners and not for limiting the protection scope of the present invention.

[0085] When the embodiments give a numerical range, it should be understood that unless otherwise specified in the present invention, any value between the two endpoints of each numerical range and the two endpoints themselves can be selected.

[0086] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the art of the present technology.

[0087] Except for the specific methods, devices, and materials used in the embodiments, according to the knowledge of those skilled in the art of the present technology and the description of the present invention, any methods, devices, and materials of the prior art similar or equivalent to those described in the embodiments of the present invention can also be used to implement the present invention.

[0088] Unless otherwise specified, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt the conventional techniques in the art of the present technology.

[0089] In animal experiments, all reagent materials and test substances used are obtained through regular commercial purchases.

[0090] Efficacy experiment of 2'-fucosyllactose and lacto-N-neotetraose composition in improving intestinal inflammatory symptoms and / or microbial dysbiosis

[0091] In this experiment, fecal samples of healthy infants aged 3 - 6 months were collected, and an infant microbiota humanized rat model was constructed using fecal transplantation technology. The rats were divided into 5 groups: normal rat control group (NC group, gavaged with 1 mL of PBS daily), model group (LPS group, gavaged with 1 mL of PBS daily), 2’-FL group (gavaged with 1 mL of 2’-FL daily, dose 443 mg / Kg body weight), LNnT group (gavaged with 1 mL of LNnT daily, dose 221.5 mg / Kg body weight), and 2’-FL + LNnT group (gavaged with 1 mL of the mixed solution of 2′-FL and LNnT daily, with a mass ratio of 2:1, total daily dose 665 mg / Kg body weight), with 12 rats in each group. After 14 days of gavage, the body weights of the rats in each group were measured. Except for the NC control group, the other 4 groups of rats were intervened with LPS (8 mg / Kg body weight), and the body weights were measured again after continuous observation for 6 h. After the animal experiment, blood samples of the rats, as well as the terminal ileum and colon tissues, were collected. The activities of MPO, the contents of DAO and D-lactic acid in the rat serum were detected using an ELISA kit (Solarbio, BC5715) according to the instructions.

[0092] The total RNA of the colon tissue was extracted after grinding and homogenizing in liquid nitrogen. The extracted RNA was reverse transcribed into cDNA using a reverse transcription kit (Qiagen, 51340). The expression levels of MUC2, ZO-1, Claudin-1, and Claudin-2 in the colon tissue were determined using a PCR instrument.

[0093] The expressions of CD4 and CD8 in the rat intestine were evaluated by immunofluorescence analysis, and the CD4 / CD8 ratio was calculated.

[0094] Finally, the total DNA was extracted using a genomic DNA extraction kit and sequenced on an Illumina MiSeq high-throughput sequencer. The data of each sample were split from the downloaded data according to the Barcode sequence and PCR amplification primer sequence. After truncating the Barcode and primer sequences, the FLASH software was used for sequence splicing. The results related to the intestinal flora composition of each group were obtained.

[0095] The group information is shown in Table 1:

[0096] Table 1

[0097] Group Details NC group Normal rat group, gavaged with PBS, without LPS treatment LPS group Model animal group, gavaged with PBS, with LPS treatment 2’-FL Monomer intervention group, gavaged with 2’-FL, with LPS treatment LNnT Monomer intervention group, gavaged with LNnT, with LPS treatment 2’FL + LNnT The ratio of 2’FL + LNnT is 2:1, gavaged with the composition, with LPS treatment

[0098] Experimental results:

[0099] 1. Changes in the body weights of rats in each group before and after LPS treatment

[0100] The body weights of all experimental rats were measured before the start of LPS injection, and there were no significant differences among the groups. The results are as Figure 1As shown in the initial body weight part. Six hours after LPS injection, the body weights of the rats in the injection groups (LPS group, 2’-FL group, LNnT group, and 2’-FL+LNnT group) were significantly decreased compared with those of the non-injected rats (NC group), and the results were as Figure 2 shown.

[0101] 2. Myeloperoxidase (MPO) activity in the serum of each group

[0102] Myeloperoxidase (MPO) in the serum is a leukocyte enzyme, mainly secreted by activated macrophages, neutrophils, and monocytes. When the body is under oxidative stress, such as infection or fever, the body can release a large amount of MPO, generate oxygen free radicals, promote the oxidation of low-density lipoprotein in the body, and promote the occurrence and development of inflammatory reactions.

[0103] The results were as Figure 3 shown. Compared with the NC group, after LPS treatment, the activity of MPO in the serum of rats in the LPS group increased significantly (p<0.05), indicating that LPS treatment would cause an increase in the level of intestinal inflammation. Compared with the LPS group, the serum MPO activities of the 2’-FL, LNnT, and 2’-FL+LNnT groups were significantly decreased. Compared with the other two groups, the 2'-FL+LNnT group could better reduce the activity of MPO in the serum, showing a certain synergistic effect, indicating that the 2'-FL+LNnT group could reduce the damage of inflammation to the intestine by reducing the MPO activity.

[0104] 3. D-lactate and diamine oxidase (DAO) levels in the serum of each group

[0105] D-lactate (D-LA) is a metabolite of intestinal bacteria, which can be produced by a variety of bacteria. Under normal circumstances, it is rarely absorbed by the human body. When the intestinal epithelial cells are damaged, the intestinal wall permeability increases, and a large amount of D-LA enters the blood. Since mammals lack the relevant enzymes to degrade D-LA, an increase in the level of D-LA in the blood can indicate damage to the intestinal barrier function.

[0106] Diamine oxidase (DAO) is a structural enzyme specifically present in small intestinal mucosal epithelial cells. It is the key enzyme for catalyzing the oxidation of histamine, and more than 90% of it exists in the small intestinal mucosa epithelium. The main role of DAO is to control the proliferation of the intestinal mucosa, and its activity is closely related to the metabolism of small intestinal mucosal epithelial cells. When the intestine is damaged or inflamed, the synthesis of DAO in the damaged area increases, and DAO enters the blood accordingly. DAO exists stably in the serum, so it can reflect the situation of intestinal barrier damage and repair.

[0107] As Figure 4A and Figure 4BAs shown, compared with the NC group, the serum D-Lactate and DAO levels of rats in the LPS group were significantly increased, indicating that the intestinal barrier of rats was severely damaged; compared with the LPS model group, the serum D-Lactate and DAO levels of rats in the intervention group were significantly decreased, indicating that their intestinal injuries were protected to a certain extent. The 2’-FL+LNnT composition was superior to 2’-FL and LNnT monomers in reducing the increase in DAO caused by LPS treatment, and there was no significant difference from the NC group, showing a certain synergistic effect.

[0108] 4. Gene expression levels of ileal mucin and tight junction proteins

[0109] Mucin is mainly composed of protein molecules, glycosyl compounds, amino acid residue modifiers, and metal ions, etc., and is one of the main components of the intestinal mucosal barrier. Members of the intestinal mucin family, such as mucin 1 (MUC1), mucin 2 (MUC2), mucin 5B (MUC5B), and mucin 13 (MUC13), are expressed in the intestine and play different functions. MUC2 consists of 5,100 amino acids and provides a protective barrier between the intestinal epithelial surface and the intestinal lumen. In addition to the mucosal barrier, tight junctions are composed of different occludin, claudin, and zonula occludens (ZOs). These three types of proteins and the actin cytoskeleton connect adjacent cells to form an epithelial barrier.

[0110] The mRNA expression levels of MUC2, Claudin-1, Claudin-2, and ZO-1 reflect the function of the intestinal barrier. The results are as Figures 5A - 5D shown. Compared with the NC group, the mRNA expression levels in the LPS group were significantly decreased (P<0.05); the 2’-FL, LNnT, and 2’-FL+LNnT groups significantly changed this result and upregulated the mRNA expression levels of MUC2, Claudin-1, Claudin-2, and ZO-1. In particular, in the 2'-FL+LNnT group, the mRNA expression levels of MUC2, Claudin-1, Claudin-2, and ZO-1 were all significantly increased, showing the most obvious effect among all intervention groups, indicating a certain synergistic effect.

[0111] 5. Expression of immune cells in each group

[0112] For the expression of immune cells in each group, see Figures 6A - 6CCompared with the NC group, the expression of CD4+ in the intestines of rats in the LPS group was significantly decreased, while the expression of CD4+ T cells in rats in the 2’-FL, LNnT, and 2’-FL+LNnT groups was significantly increased. Compared with the NC group, the expression of CD8+ T cells in the LPS group was significantly increased, while the expression of CD8+ T cells in rats in the 2’-FL, LNnT, and 2’-FL+LNnT groups was significantly decreased. The CD4+ / CD8+ T results showed that compared with the NC control group, the ratio of CD4+ / CD8+ T cells in the LPS group was significantly decreased (P<0.05), indicating that the proportion of intestinal immune cells in rats was dysregulated and the immune function was impaired after LPS treatment. In the 2’-FL, LNnT, and 2’-FL+LNnT groups, the CD4+ / CD8+ was significantly increased, especially in the 2'-FL+LNnT group, which showed a significant increase in the ratio of CD4+ / CD8+ T cells, indicating a certain coordination effect.

[0113] 6. Effect of the combination of 2’-FL and LNnT on intestinal flora

[0114] In this experiment, the effect of the 2’-FL+LNnT composition on the intestinal flora of infants and young children was further studied.

[0115] As Figure 7 shown, the specific OTUs in the NC group were significantly more than those in the LPS group, indicating that LPS treatment led to a significant reduction in the species of intestinal microorganisms in rats. The number of OTUs in the 2'-FL+LNnT group was restored, indicating that the 2’-FL+LNnT group could significantly improve the reduction in the species of intestinal microorganisms caused by intestinal mucosal immune injury.

[0116] Table 2 shows the results of α-diversity. The results show that compared with the NC group, the observed species index, chao1 index, shannon index, and simpson index in the LPS group were all significantly decreased. Compared with the LPS group, the four indexes in the 2'-FL+LNnT group were all significantly increased. The results indicate that 2'-FL+LNnT can significantly improve the diversity and richness of the microbial community.

[0117] Table 2. α-diversity analysis of each group

[0118]

[0119]

[0120] Using the 16s sequencing method, the flora composition of intestinal microorganisms was analyzed. See the results of the flora composition analysis in Figure 8. The results showed that compared with the NC control group, the abundances of Lactobacillus and Muribaculaceae in the LPS group decreased significantly by 9.09% and 8.11% respectively, while the abundances of Escherichia-Shigella and Alloprevotella increased significantly by 7.15% and 11.14% respectively. Compared with the LPS model group, the abundance of Muribaculaceae in the 2'-FL+LNnT group increased by 4.11%, the abundance of Escherichia-Shigella decreased significantly by 4.17%, and the abundance of Alloprevotella decreased significantly by 19.40%. Muribaculaceae is a newly established genus name for this bacterium, which was formerly known as the S24-7 family and plays an important role in the process of fermenting carbohydrates. From the results of the flora, it can be seen that the 2'-FL+LNnT group successfully improved the structure of the intestinal flora compared with the LPS model group, especially reducing the abundances of some potentially harmful genera such as Escherichia-Shigella and Alloprevotella, and increasing the abundances of some potentially beneficial genera such as Muribaculaceae and Ruminococcus. The changes in other genera included some minor adjustments, making the flora structure of the 2'-FL+LNnT group closer to that of the NC control group.

[0121] Example 1

[0122] This example provides a powdered infant formula for infants aged 0 to 6 months. The total protein content in the powdered formula is 10.5 g / 100 g powder, the fat content is 27 g / 100 g powder; the carbohydrate content is 54.2 g / 100 g powder, 2’-FL is 1125 mg / 100 g, and LNnT is 375 mg / 100 g powder. The ratio of 2’-FL:LNnT is 3:1.

[0123] This example is achieved by compounding raw materials in the following weight parts. The raw material composition includes (preparing 1000 parts by weight):

[0124] 800 parts of raw cow's milk, 300 parts of desalted whey powder (D90), 220 parts of edible vegetable blending oil, 190 parts of lactose, 35 parts of galacto-oligosaccharide, 27 parts of α-lactalbumin powder, 22 parts of casein, 8 parts of oligofructose, 5 parts of whey protein powder (rich in milk fat globule membrane), 4 parts of phospholipids, 24 parts of compound nutrients, wherein the compound nutrients include about 4 parts of compound vitamin nutrition package, about 2 parts of choline chloride nutrition package, about 12 parts of mineral 2 nutrition package, about 1 part of mineral 1 nutrition package, about 2 parts of magnesium chloride nutrition package and about 3 parts of potassium chloride nutrition package, and the base materials of each nutrition package are lactose, 12 parts of 2'-fucosyllactose (2'-FL), 4 parts of lactose-N-neotetraose (LNnT), 16 parts of DHA, 22 parts of ARA, about 7 parts of lactoferrin and about 0.7 parts of nucleotides.

[0125] The preparation process of the infant formula milk powder of this embodiment:

[0126] The mixed material is homogenized after filtering and removing impurities, and the homogenization temperature is not less than 50°C and the pressure is not less than 120bar. Subsequently, the homogenized material is concentrated, and the dry matter weight content of the concentrate is 40% to 60%. The concentrated material is sterilized at above 80°C for 10 to 60 seconds. The sterilized material is dried by a spray drying tower (inlet air temperature 150°C to 200°C, exhaust air temperature 80°C to 120°C, negative pressure -10 to -2mmWG, high pressure pump pressure 140 to 250bar), and then introduced into a fluidized bed for secondary drying to obtain a powder.

[0127] After testing, all indicators of the finished product meet the standard requirements for infant formula milk powder.

[0128] Example 2

[0129] The present embodiment provides a powdered formula food for infants aged 0 to 6 months, wherein the total protein content of the powdered formula food is 10.5 g / 100 g powder, the fat content is 27 g / 100 g powder, the carbohydrate content is 54.6 g / 100 g powder, the 2'-FL is 590 mg / 100 g powder, and the LNnT is 285 mg / 100 g powder, and the ratio of 2'-FL:LNnT is 2:1.

[0130] This embodiment is achieved by compounding the following raw materials in parts by weight, and its raw material composition includes (preparing 1000 parts by weight):

[0131] 1000 parts of raw cow milk, 360 parts of demineralized whey powder (D90), 230 parts of edible vegetable blended oil, 170 parts of lactose, 30 parts of galactooligosaccharide, 25 parts of α-lactalbumin powder, 20 parts of casein, 8 parts of fructooligosaccharide, 5 parts of whey protein powder (rich in milk fat globule membrane), 4 parts of phospholipid, 24 parts of compound nutrients, among which the compound nutrients include about 4 parts of compound vitamin nutrient package, about 2 parts of choline chloride nutrient package, about 12 parts of mineral two nutrient package, about 1 part of mineral one nutrient package, about 2 parts of magnesium chloride nutrient package and about 3 parts of potassium chloride nutrient package, the base material of each nutrient package is lactose, 6 parts of 2'-fucosyllactose (2'-FL), 3 parts of lacto-N-neotetraose (LNnT), 13 parts of DHA, 15 parts of ARA, and about 1 part of lactoferrin.

[0132] The specific preparation process of the infant formula milk powder in this example is the same as that in Example 1.

[0133] After testing, the various indicators of this finished product meet the standard requirements of infant formula milk powder.

[0134] Example 3

[0135] This example provides a powdered infant formula food for 0-6 months old. The total protein content in this powdered formula food is 10.5 g / 100 g powder, the fat content is 27 g / 100 g powder; the carbohydrate content is 52.2 g / 100 g powder, 2'-FL is 1475 mg / 100 g powder, LNnT is 295 mg / 100 g powder, and the ratio of 2'-FL:LNnT is 5:1.

[0136] This example is achieved by compounding the following raw materials in parts by weight. Its raw material composition includes (preparing 1000 parts by weight):

[0137] 1500 parts of raw cow milk, 490 parts of demineralized whey powder (D90), 200 parts of edible vegetable blended oil, 20 parts of lactose, 30 parts of galactooligosaccharide, 15 parts of fructooligosaccharide, 5 parts of whey protein powder (rich in milk fat globule membrane), 4 parts of phospholipid, 24 parts of compound nutrients, among which the compound nutrients include about 4 parts of compound vitamin nutrient package, about 2 parts of choline chloride nutrient package, about 12 parts of mineral two nutrient package, about 1 part of mineral one nutrient package, about 2 parts of magnesium chloride nutrient package and about 3 parts of potassium chloride nutrient package, the base material of each nutrient package is lactose, 15 parts of 2'-fucosyllactose (2'-FL), 3 parts of lacto-N-neotetraose (LNnT), 10 parts of DHA, 13 parts of ARA, and about 0.4 part of nucleotide.

[0138] The specific preparation process of the infant formula milk powder in this example is the same as that in Example 1.

[0139] After testing, the various indicators of this finished product meet the standard requirements of infant formula milk powder.

[0140] The above description of the embodiments is to enable those of ordinary skill in the art to understand and use the invention. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative efforts. For example, infant formula liquid milk can be converted according to the dilution ratio with reference to the formula range of infant formula milk powder. Therefore, the present invention is not limited to the above embodiments, and the improvements and modifications made by those skilled in the art without departing from the scope of the present invention according to the disclosure of the present invention should be within the protection scope of the present invention.

Claims

1. A pediatric formula food, the pediatric formula food being an infant formula food or a children's formula food, the pediatric formula food comprising 2'-fucosyllactose and lactose-N-neotetraose, the mass ratio of 2'-fucosyllactose to lactose-N-neotetraose being (1-5.5):1; in, The infant formula food has a total protein content of 7.8g to 22.8g / 100g, a fat content of 14.9g to 33.8g / 100g, and a carbohydrate content of 40.7g to 79.3g / 100g, converted into dry matter; the children's formula food has a total protein content of 16.5g to 30g / 100g, a fat content of 10g to 35g / 100g, and a carbohydrate content of 40g to 70g / 100g, converted into dry matter.

2. The pediatric formula food according to claim 1, wherein: The mass ratio of 2'-fucosyllactose to lactose-N-neotetraose is (1-4.8):1, preferably (1-4):1; More preferably, the mass ratio of 2'-fucosyllactose to lacto-N-neotetraose is 2:

1.

3. The pediatric formula food according to claim 1 or 2, wherein: The amount of 2'-fucosyllactose contained in the food is 0.5-1.8 g / 100 g of food dry matter.

4. The pediatric formula food according to claim 1 or 2, wherein: The amount of 2'-fucosyllactose in the food is 0.6-0.7g / 100g food dry matter, 0.8-0.9g / 100g food dry matter, 1.0-1.1g / 100g food dry matter, 1.2-1.3g / 100g food dry matter, 1.4-1.5g / 100g food dry matter or 1.6-1.7g / 100g food dry matter.

5. The pediatric formula food according to claim 1, wherein the raw materials thereof comprise: Raw milk: 700-3700 servings; Whey powder: 0-600 parts; Whey protein powder: 0-100 servings; Edible vegetable blended oil: 80-240 parts; Lactose: 0-550 parts; Minerals: 8-30 parts; Trace elements: 0.5-2 parts; Vitamins: 2.5-10 portions; α-lactalbumin: 0.625-3 parts; Linoleic acid: 2.5-5 parts; 2'-fucosyllactose raw material: 2.5-13 parts; Lactose-N-neotetraose raw material: 2.5-5 parts.

6. The pediatric formula food according to any one of claims 1 to 5, which is milk powder or liquid milk.

7. A method for preparing the pediatric formula food according to any one of claims 1 to 6, the method comprising: The raw materials providing 2'-fucosyllactose and lactose-N-neotetraose are mixed with other raw materials in the formula by wet or dry method or dry-wet composite production process, and the mixed materials are homogenized after filtering and removing impurities, and are further used to prepare the pediatric formula food.

8. The preparation method according to claim 7, further comprising: The homogenized material is concentrated to a dry matter weight content of 40% to 60%; The concentrated material is sterilized at above 80°C for 10 to 60 seconds; The sterilized material is dried by a spray drying tower (inlet air temperature 150°C ~ 200°C, exhaust air temperature 80°C ~ 120°C, negative pressure -10 ~ -2mmWG, high pressure pump pressure 140 ~ 250bar), and then introduced into a fluidized bed for secondary drying to obtain powder.

9. Use of the pediatric formula food according to any one of claims 1 to 6 in the preparation of a product for improving intestinal inflammation symptoms and / or microbial imbalance.

10. The use according to claim 9, wherein: The improvement of intestinal inflammation symptoms and / or microbial imbalance includes one or more of the following: Improve weight loss caused by intestinal inflammatory response; Improve intestinal damage caused by inflammatory response; Reduce the increase in serum myeloperoxidase (MPO) activity caused by intestinal inflammatory response; Reduce the increase in serum D-lactic acid levels caused by intestinal inflammatory response; Reduce the increase in serum diamine oxidase (DAO) levels caused by intestinal inflammatory response; Improve intestinal barrier function; strengthening the intestinal physical and / or mucus barrier; Increase the expression level of intestinal mucin; Increase the expression levels of intestinal occludin, claudin and / or zonulaoccludens (ZOs); Maintain intestinal cell immune homeostasis; Improve intestinal immune cell disorders caused by inflammatory response; Increase intestinal CD4+T cell expression; Reduce intestinal CD8+T cell expression; Improve the ratio of intestinal CD4+ / CD8+T cells; Improving the diversity and / or richness of the intestinal microbiome; Restoring the disturbance or reduced diversity of the intestinal cornerstone flora caused by inflammation or antibiotics; Improve the reduction of intestinal microbial species caused by intestinal mucosal immune damage; Increased abundance of the intestinal bacterial genera Muribaculaceae and / or Ruminococcus; Reduction of the abundance of intestinal Escherichia-Shigella and / or Alloprevotella.

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