Use of a nutritional composition

Abstract

The use of a nutritional composition is disclosed. The nutritional composition comprises medium- and long-chain fatty acid triglycerides and nervonic acid in a mass ratio of 1:1 to 5000:1. This nutritional composition has a synergistic effect in improving immunity, particularly in restoring the number of one or more macrophages, neutrophils, and T cells in immunocompromised individuals.

Description

Technical Field

[0001] This application relates to the fields of nutrition, food science, and pharmaceutical technology, and in particular to the use of nutritional compositions in the preparation of products for improving immunity. Background Technology

[0002] Immunity is the protective physiological response of the body to distinguish between "self" and "non-self" and to eliminate antigens and foreign substances. Its core lies in maintaining homeostasis (Peng Wanqiang, Immunity and Immune Regulation (II) [J]. Guangdong Animal Husbandry and Veterinary Science and Technology, 1984, (2): 57-62). As a key line of defense for the body's health, the immune system, once its function is impaired, will significantly increase the host's susceptibility to diseases such as infection and cancer. In the innate immune response, neutrophils, as the most abundant phagocytes, can quickly chemotactically approach the site of infection and efficiently eliminate pathogens through phagocytosis, degranulation, and the release of neutrophil extracellular traps. At the same time, macrophages also have a strong phagocytic function, not only directly phagocytizing and eliminating pathogens, but also bridging innate immunity and adaptive immunity through antigen presentation and secretion of cytokines, and precisely regulating inflammatory responses. On the adaptive immune side, T cells, as the core executors, play an irreplaceable role in mediating cellular immunity and assisting humoral immunity. Neutrophils, macrophages, and T cells work together to form an important pillar of the human body's systemic immune defense system.

[0003] The role of dietary lipids in nutrition and immune regulation is receiving increasing attention. Medium- and long-chain triacylglycerols (MLCTs) are a novel type of structural lipid that combines medium- and long-chain fatty acids on a glycerol backbone. The fatty acid composition of MLCTs is closer to that of breast milk, which is beneficial for fat digestion and absorption, enhancing the absorption of lipid nutrients, inhibiting body fat accumulation, and providing rapid and stable energy (see CHENG X et al., “Medium- and Long-Chain Triacylglycerol: Preparation, Health Benefits, and Food Utilization”, Annual Review of Food Science and Technology, 2024, 15: 381-408.). Compared to physical mixtures of long-chain triacylglycerols (LCTs) and medium-chain triacylglycerols (MCTs) with the same overall fatty acid composition, MLCTs exhibit different metabolic characteristics and physiological functions. Animal experiments have confirmed that, under conditions of consistent overall fatty acid composition, feeding mice with formulated lipid diets containing long-chain triglycerides with a human milk-like structure, compared to diets containing physically mixed long-chain triglycerides / medium-chain triglycerides from vegetable oils, can regulate thermogenesis, inhibit visceral fat accumulation, alter gut microbiota structure, and regulate lipid metabolism (Yuan, Tinglan et al., “Novel Human Milk Fat Substitutes Based on Medium- and Long-Chain Triacylglycerol Regulate Thermogenesis, Lipid Metabolism, and Gut Microbiota Diversity in C57BL / 6J Mice.” Journal of Agricultural and Food Chemistry, vol. 72, no. 12, 2024, pp. 6213-6225.).

[0004] Patent application WO 2024 / 146188 A1 discloses a fat component for a special medical purpose formulation food. In its fatty acid composition, medium-chain fatty acids account for 20 wt% to 50 wt% of the total fatty acids, the mass ratio of medium-chain to long-chain fatty acids is 1:1 to 1:3, and in its triglyceride structure, medium- and long-chain triglycerides account for 30% to 85 wt%, medium-chain fatty acids at the sn-1 and 3 positions account for 30 to 60 wt%, and long-chain fatty acids at the sn-2 position account for 65 wt% to 90 wt%. This fat component has a higher digestibility and absorption rate, can improve the bioavailability of EPA and DHA, helps improve the nutritional status of liver cancer patients during chemotherapy, provides immediate energy for postoperative liver cancer patients, and reduces inflammatory responses and improves immune function.

[0005] On the other hand, nervonic acid (NA), chemically known as cis-15-docosahexaenoic acid, is an omega-9 monounsaturated fatty acid that helps repair damaged nerve cells and tissues and promotes brain development. It holds significant research value in the prevention and treatment of brain diseases. The nervonic acid synthesized in the human body cannot fully meet the body's health needs, especially for special populations such as newborns, who require additional intake from external sources. However, its role in immunity remains unclear.

[0006] There is still a need in this field for nutritional compositions that can improve immunity. Summary of the Invention

[0007] To address the aforementioned technical problems, this application investigated the effect of a combination of medium- and long-chain fatty acid triglycerides and nervonic acid on improving immunity. It was found that a combination of medium- and long-chain fatty acid triglycerides and nervonic acid at a specific mass ratio (e.g., 0.1:1 to 5000:1, preferably 1:1 to 2500:1) can synergistically improve immunity and enhance immune efficacy.

[0008] In one aspect, this application provides the use of a nutritional composition for non-therapeutic purposes to improve immunity, characterized in that the nutritional composition comprises medium- and long-chain fatty acid triglycerides and nervonic acid, and the mass ratio of the medium- and long-chain fatty acid triglycerides to the nervonic acid is in the range of 0.1:1 to 5000:1.

[0009] In some implementations, the improvement of immunity includes enhancing systemic immunity and / or restoring one or more of the macrophages, neutrophils, and T cells that are reduced in immunocompromised individuals.

[0010] In some embodiments, the mass ratio of the medium- and long-chain fatty acid triglycerides to the nervonic acid is in the range of 1:1 to 2500:1.

[0011] In some embodiments, the mass ratio of the medium- and long-chain fatty acid triglycerides to the nervonic acid is in the range of 1.5:1 to 2400:1.

[0012] In some embodiments, the nutritional composition is used in the form of a food comprising the nutritional composition.

[0013] In some embodiments, the food is a health food that helps boost immunity.

[0014] In some embodiments, the food is infant food.

[0015] In some embodiments, the food is selected from one or more of dairy products, confectionery, beverages, bread, and biscuits.

[0016] In some embodiments, the food is milk powder or fermented food.

[0017] In some embodiments, the milk powder is selected from one or more of the following: infant formula, children's formula, adolescent formula, pregnant women's formula, postoperative patient formula, and middle-aged and elderly formula.

[0018] On the other hand, this application also provides the use of a nutritional composition in the preparation of a product, characterized in that the product is used to improve immunity, the nutritional composition comprising medium- and long-chain fatty acid triglycerides and nervonic acid, and the mass ratio of the medium- and long-chain fatty acid triglycerides to the nervonic acid is in the range of 0.1:1 to 5000:1.

[0019] In some embodiments, the mass ratio of the medium- and long-chain fatty acid triglycerides to the nervonic acid is in the range of 1:1 to 2500:1.

[0020] In some embodiments, the mass ratio of the medium- and long-chain fatty acid triglycerides to the nervonic acid is in the range of 1.5:1 to 2400:1.

[0021] In some embodiments, the product is a pharmaceutical product or a health food product that helps improve immunity.

[0022] The beneficial effects of this application include: when medium- and long-chain fatty acid triglycerides and nervonic acid are used in combination at a mass ratio of 0.1:1 to 5000:1, preferably 1:1 to 2500:1, and more preferably 1.5:2400, there is a synergistic effect between the two in restoring the number of one or more of macrophages, neutrophils, and T cells reduced in immunocompromised individuals. Compositions containing medium- and long-chain fatty acid triglycerides and nervonic acid at a mass ratio of 0.1:1 to 5000:1 can be used to prepare pharmaceuticals for improving immunity or health foods that help enhance immunity. Attached Figure Description

[0023] To enable a full understanding of this application and its easy implementation, exemplary embodiments will now be described by way of non-limiting examples, with reference to the accompanying drawings. In the drawings:

[0024] Figure 1 A typical fluorescence intensity diagram of zebrafish T cells after sample treatment is shown, in which the red dashed line represents the analysis area;

[0025] Figure 2 A typical fluorescence intensity diagram of zebrafish neutrophils after sample treatment is shown, in which the yellow dashed line represents the analysis area;

[0026] Figure 3 A typical fluorescence intensity diagram of zebrafish macrophages after sample treatment is shown, in which the yellow dashed line represents the analysis area. Detailed Implementation

[0027] Unless otherwise indicated or defined, all terms used have their ordinary meaning in the art as would be understood by those skilled in the art. Furthermore, unless otherwise stated, all methods, steps, techniques, and operations not specifically detailed herein can and have been performed in a manner known per se as would be understood by those skilled in the art.

[0028] the term

[0029] In this application, the terms "medium- and long-chain triacylglycerol" or "MLCT" are used interchangeably to refer to triacylglycerols with a special structure in which both medium-chain and long-chain fatty acid chains are present on the backbone. As is known in the art, medium- and long-chain triacylglycerols are produced from edible vegetable oils and medium-chain triacylglycerols through transesterification with lipases, followed by distillation, decolorization, deodorization, and other processes. Medium- and long-chain triacylglycerols have attracted widespread attention as a star product among novel structural lipids. Their fatty acid composition is close to that of breast milk, which is beneficial for fat digestion and absorption. They can improve the absorption of lipid nutrients, inhibit the accumulation of body fat, and provide rapid and stable energy (Xinyi Cheng et al., Medium- and Long-Chain Triacylglycerol: Preparation, Health Benefits, and Food Utilization, *Annu. Rev. Food Sci. Technol.*, 2024, 15:381-408). Medium- and long-chain triacylglycerols have different metabolic characteristics and physiological functions than physically mixed long-chain and medium-chain triacylglycerols.

[0030] In this application, the term "medium-chain fatty acid" refers to fatty acids with 6 to 12 carbon atoms in their carbon chain, such as hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, and dodecanoic acid. The term "long-chain fatty acid" refers to fatty acids with 14 or more carbon atoms in their carbon chain, such as 14 to 30 carbon atoms, such as myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, and nervonic acid. The terms "medium-chain fatty acid triglyceride" and "long-chain fatty acid triglyceride" refer to the esterification products of "medium-chain fatty acid" and "long-chain fatty acid" with glycerol, respectively. The term "medium- and long-chain fatty acid triglycerides" refers to triglyceride compounds that simultaneously contain both medium-chain fatty acid residues and long-chain fatty acid residues in their molecular structure. This application does not have specific requirements for the medium- and long-chain fatty acid triglycerides used; those commonly used in the art can be used. Medium- and long-chain triglycerides can be used in pure form or in a non-pure form rich in medium- and long-chain triglycerides.

[0031] For example, the medium- and long-chain fatty acid triglycerides used in this application may contain C6~C6. 12 fatty acid residues and C 14 ~C 30 Fatty acid residues. C6~C 12 Fatty acid residues can originate from one or more of the following: hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, etc. C 14 ~C 30 The fatty acid residues may be derived from one or more of the following: myristic acid, palmitic acid, heptadecanic acid, oleic acid, linoleic acid, stearic acid, nonadecanic acid, eicosapentaenoic acid, docosahexaenoic acid, docosapatraenoic acid, docosahexaenoic acid, tricarboxylic acid, tetracarboxyenoic acid, docosapentaenoic acid, docosahexaenoic acid, arachidonic acid, etc. In some embodiments, C6~C 12 The fatty acid residues are derived from one or more of the following: hexanoic acid, octanoic acid, decanoic acid, and lauric acid. In some embodiments, C 14 ~C 30 The fatty acid residues are derived from one or more of the following: myristic acid, palmitic acid, oleic acid, linoleic acid, stearic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid. In some embodiments, C6~C 12 Fatty acid residues and C 14 ~C 30 The mass ratio of fatty acid residues is from 0.124 to 2.000. In some embodiments, C6~C 12 Fatty acid residues on the C 14 ~C 30The mass ratio of fatty acid residues can be within the range defined by 0.124, 0.125, 0.130, 0.140, 0.150, 0.160, 0.170, 0.180, 0.190, 0.200, 0.300, 0.400, 0.500, 0.600, 0.700, 0.800, 0.900, 1.000, 1.100, 1.200, 1.300, 1.400, 1.500, 1.600, 1.700, 1.800, 1.900, 2.000, or any two of these. In some embodiments, the "medium- and long-chain fatty acid triglycerides" can be prepared according to the method of patent application CN115369132A.

[0032] In this application, the term "nervonic acid," also known as "squalane," refers to an ω-9 monounsaturated fatty acid with the chemical name cis-15-tetracosanoic acid. Traditionally, nervonic acid is believed to promote nerve development and myelin formation, repair nerve damage, and delay neurodegenerative diseases, but its effects on immunity are not yet fully understood.

[0033] To date, further research is needed to determine whether combining medium- and long-chain triglycerides with nervonic acid can synergistically improve immunity and enhance immune efficacy.

[0034] In this application, the term "immunity" refers to the sum of biological effects produced by the body in recognizing and responding to "self" and "foreign (non-self)" entities, which is a physiological function that maintains homeostasis under normal circumstances. This process is carried out by the immune system, which consists of multiple parts, including immune organs (such as bone marrow, thymus, lymph nodes, spleen, etc.), immune cells (such as T lymphocytes, B lymphocytes, phagocytes, natural killer cells, etc.), and immune molecules (such as antibodies, complement, cytokines, etc.). The immune system has three core physiological functions: immune defense (resisting the invasion of pathogenic microorganisms), immune surveillance (identifying and eliminating mutated cells, tumor cells, and virus-infected cells in the body), and immune homeostasis (eliminating aging and dead cells to maintain homeostasis). The term "immunity" refers to the comprehensive ability of the immune system to perform the above functions, that is, the body's intrinsic ability to protect itself and resist "non-self". In layman's terms, immunity is the body's ability to recognize and eliminate any foreign invaders (such as viruses and bacteria), process aging, damaged, dead, and degenerated cells, and recognize and deal with mutated cells and virus-infected cells. The strength of immunity is directly reflected in the body's ability to resist disease. Under the same pathogen exposure conditions, individuals with strong immunity may not develop symptoms or may experience mild symptoms, while individuals with weak immunity are more susceptible and experience more severe symptoms.

[0035] The immune system, a crucial line of defense for the body's health, significantly increases the host's susceptibility to infections and diseases such as cancer when its function is impaired. In the innate immune response, neutrophils, the most abundant phagocytes, rapidly chemotactically approach the site of infection, efficiently clearing pathogens through phagocytosis, degranulation, and the release of neutrophil extracellular traps. Simultaneously, macrophages also possess powerful phagocytic functions, not only directly engulfing and clearing pathogens but also bridging innate and adaptive immunity through antigen presentation and cytokine secretion, and precisely regulating inflammatory responses. On the adaptive immune side, T cells, as the core executors, play an irreplaceable role in mediating cellular immunity and assisting humoral immunity. Neutrophils, macrophages, and T cells work synergistically to form a vital pillar of the body's systemic immune defense system.

[0036] In this application, the term "non-therapeutic purpose" has the conventional meaning in the art, including, for example, nutritional and / or health purposes, such as for the preparation of food such as ordinary food, functional food or health food.

[0037] In this application, the term "improved immunity" refers to restoring abnormal immune function to a healthy balance through nutritional or pharmacological interventions. For example, for individuals with weakened immunity (such as those undergoing chemotherapy or post-operative care for cancer patients), it is necessary to enhance immune defense and surveillance capabilities; for those with overactive or disordered immune systems (such as those with autoimmune diseases), it is necessary to suppress excessive inflammatory responses; for infants and young children whose immune systems are still in the "learning" and "shaping" stage, it is necessary to provide them with the necessary nutrients for growth and development, activate their immune instincts, enhance immune memory, and create an environment suitable for the healthy development of their immune system. In some aspects, improving immunity may include enhancing systemic immunity and / or restoring one or more of the macrophages, neutrophils, and T cells that are reduced in individuals with weakened immunity.

[0038] Nutritional compositions and their uses

[0039] In one aspect, this application provides the use of a nutritional composition for non-therapeutic purposes to improve immunity, said nutritional composition comprising medium- and long-chain fatty acid triglycerides and nervonic acid, wherein the mass ratio of medium- and long-chain fatty acid triglycerides to nervonic acid is from 0.1:1 to 5000:1, for example from 1:1 to 2500:1, or even from 1.5:1 to 2400:1.

[0040] In another aspect, this application provides a method for improving immunity, comprising administering a nutritional composition to a subject, the nutritional composition comprising medium- and long-chain fatty acid triglycerides and nervonic acid, wherein the mass ratio of medium- and long-chain fatty acid triglycerides to nervonic acid is from 0.1:1 to 5000:1, for example from 1:1 to 2500:1, for example from 1.5:1 to 2400:1.

[0041] In another aspect, this application provides a product for improving immunity, the product comprising the nutritional composition described in any embodiment herein, the nutritional composition comprising medium- and long-chain fatty acid triglycerides and nervonic acid, and the mass ratio of medium- and long-chain fatty acid triglycerides to nervonic acid being 0.1:1 to 5000:1, for example 1:1 to 2500:1, for example 1.5:1 to 2400:1.

[0042] In any embodiment of this application, the improvement of immunity includes enhancing systemic immunity and / or restoring one or more of the macrophages, neutrophils and T cells that are reduced in immunocompromised individuals.

[0043] In any embodiment of this application, the mass ratio of medium- and long-chain fatty acid triglycerides to nervonic acid in the nutritional composition may be any value selected from the group consisting of or within any two of these values: about 0.1:1, about 0.3:1, about 0.5:1, about 0.8:1, about 1:1, about 1.2:1, 1.3:1, about 1.4:1, about 1.5:1, about 1.6:1, about 1.7:1, about 1.8:1, about 1.9:1, about 2:1, about 2.2:1, about 2.5:1, about 2.8:1, about 3:1, about 3.2:1, approximately 3.5:1, approximately 3.8:1, 4:1, approximately 4.5:1, approximately 5:1, approximately 5.5:1, approximately 6:1, approximately 7:1, approximately 8:1, approximately 9:1, approximately 10:1, approximately 12:1, approximately 15:1, approximately 18:1, approximately 20:1, approximately 25:1, approximately 30:1, approximately 35:1, approximately 40:1, approximately 45:1, approximately 50:1, approximately 55:1, approximately 60:1, approximately 70:1, approximately 80:1, approximately 90:1, approximately 100:1, approximately 110:1, approximately 120:1, approximately 150: 1. Approximately 180:1, Approximately 200:1, Approximately 220:1, Approximately 250:1, Approximately 280:1, Approximately 300:1, Approximately 320:1, Approximately 350:1, Approximately 380:1, Approximately 400:1, Approximately 420:1, Approximately 450:1, Approximately 480:1, Approximately 500:1, Approximately 520:1, Approximately 550:1, Approximately 580:1, Approximately 600:1, Approximately 650:1, Approximately 700:1, Approximately 750:1, Approximately 800:1, Approximately 800:1, Approximately 900:1, Approximately 950:1, Approximately 1000:1 Approximately 1050:1, approximately 1100:1, approximately 1200:1, approximately 1300:1, approximately 1400:1, approximately 1500:1, approximately 1600:1, approximately 1700:1, approximately 1800:1, approximately 1900:1, approximately 2000:1, approximately 2100:1, approximately 2200:1, approximately 2300:1, approximately 2400:1, approximately 2500:1, approximately 2800:1, approximately 3000:1, approximately 3500:1, approximately 4000:1, approximately 4500:1, approximately 4800:1, and approximately 5000:1. Preferably, the mass ratio of medium- and long-chain fatty acid triglycerides to nervonic acid in the nutritional composition is about 1.5:1, about 3:1, about 60:1, about 600:1, about 1200:1, or about 2400:1.

[0044] In some embodiments, the nutritional composition described in this application may be used in the form of food or pharmaceuticals containing the nutritional composition.

[0045] The medium- and long-chain triglycerides and nervonic acid in the nutritional composition may be used in amounts permitted for food science or pharmaceutical purposes. For example, the content of medium- and long-chain triglycerides may be from 0.1% to 30% by mass based on the total mass of the product. As an example, the content of medium- and long-chain triglycerides may be 0.1%, 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% by mass based on the total mass of the product, or a range defined by any two thereof. As another example, the content of nervonic acid may be from 0.002% to 0.3% based on the total mass of the product. As an example, based on the total mass of the product, the content of nervonic acid may be 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.25, 0.30%, or within the range defined by any two of these.

[0046] In some embodiments, when the nutritional composition described herein is used as a food, the food may be a general food or a specialty food. As an example, the food may be selected from dairy products, confectionery, beverages, bread, and biscuits; for instance, the food may be selected from milk powder or fermented foods.

[0047] In some implementations, the milk powder may be, for example, infant formula.

[0048] In some implementations, the food may be, for example, a functional food or health food that helps improve immunity.

[0049] Those skilled in the art will readily understand that, in addition to the nutritional composition, the food may also contain various food ingredients, such as one or more selected from the following: raw milk, demineralized whey powder, whey protein concentrate, lactose, blended vegetable oils, fructooligosaccharides, galactooligosaccharides, nucleotides, choline, vitamins, minerals, DHA, and taurine. These food ingredients are readily selected by those skilled in the art. For example, when the food is milk powder, in addition to the nutritional composition described in any embodiment herein, the milk powder may also include proteins such as α-lactalbumin and milk fat globule membrane protein; carbohydrates such as lactose; lipids; minerals such as calcium, iron, and phosphorus; vitamins; and other additives such as whey powder, choline tartrate, docosahexaenoic acid, arachidonic acid, and walnut oil.

[0050] In some embodiments, when the nutritional composition described herein is used as a pharmaceutical product, the product may also contain pharmaceutically acceptable carriers, diluents, or excipients.

[0051] In some embodiments, the product may also contain various excipients acceptable to food or pharmaceutical use (e.g., flavorings or colorings). The product may be in any suitable form, such as liquid, solid, powder, gel, etc., as long as it is suitable for use by the subject. When the product is a pharmaceutical product, it may be any dosage form, such as a solid dosage form (powder, tablet, etc.) or a liquid dosage form.

[0052] The food or medicine may be produced using preparation methods commonly used in the art, which will not be elaborated here.

[0053] In some embodiments, the nutritional composition or a product containing the nutritional composition may be used orally.

[0054] The present application will be more easily understood by referring to the following embodiments, which are only used to illustrate certain aspects and implementation methods of the present application and are not intended to limit the present application.

[0055] Example

[0056] Unless otherwise stated, all reagents used in this embodiment are commercially available or conventional materials. All of the following embodiments:

[0057] Medium- and long-chain fatty acid triglycerides (hereinafter referred to as "MLCT"): derived from medium- and long-chain fatty acid edible oils, with a purity of 69±10%, produced by Qingdao Haizhiyuan Life Technology Co., Ltd., with production batch number Y1505-22120101.

[0058] Nervonic acid (hereinafter referred to as "NA"): purity of over 90%, purchased from Guangzhou Green Extract Biotechnology, production batch number FH20240120.

[0059] The positive control drug, Bailin Capsules, was provided by Hangzhou Sino-American East China Pharmaceutical Co., Ltd., with production batch number 2411018D. Bailin Capsules are a widely used traditional Chinese medicine in clinical practice. Its main ingredient is fermented Cordyceps sinensis powder, which has the effects of enhancing immunity.

[0060] The modeling reagent, vinorelbine tartrate injection, was provided by Jiangsu Hansoh Pharmaceutical Co., Ltd., with production batch number 600211003. Existing literature indicates that vinorelbine tartrate has long-term toxic effects on the immune and hematopoietic systems of rats, manifested as thymic atrophy and bone marrow suppression.

[0061] 3-day-fiber (dpf) transgenic T-cell red fluorescent zebrafish, provided by Hangzhou Huante Bio-Fish Farming Center, were bred through natural pair mating and reached an age of 3 dpf. The T cells of this strain exhibit red fluorescence under a fluorescence microscope, used for cell quantification. The number of T cells in the thymus directly depends on the ability of hematopoietic stem cells (HSCs) to migrate from the bone marrow (head kidney in zebrafish) to the thymus, and the ability of the thymic microenvironment to support T-cell proliferation.

[0062] 3-day-fif (dpf) transgenic macrophage-producing green fluorescent zebrafish, provided by Hangzhou Huante Bio-Fish Farming Center, were bred through natural pair mating and reached 3 days post-fertilization (dpf). The macrophages of this strain exhibited green fluorescence under a fluorescence microscope, which was used for cellular quantification.

[0063] 3-day-fif (dpf) transgenic green fluorescent zebrafish with neutrophils, provided by Hangzhou Huante Bio-Fish Farming Center, were bred through natural pair mating and reached 3 days of age. Neutrophils can be specifically labeled under in vivo, and their number and activation level can be quantitatively characterized by changes in fluorescence intensity and distribution. This method is suitable for evaluating the mechanisms and efficacy of immune function regulation, protection against chemotherapy-induced immune damage, and anti-inflammatory activities.

[0064] In biomedical and food science research, the zebrafish immunodeficiency model is a standard experimental tool, primarily used for evaluating the efficacy of functional food / health product ingredients, screening drugs (especially immunomodulators or drugs to alleviate chemotherapy side effects), and studying basic immune mechanisms. By inducing immune cell development and hematopoietic function damage in zebrafish using chemosuppressive drugs, resulting in a decrease in the number of immune cells such as neutrophils, macrophages, and T cells, and a reduction in immune function, the number of immune cells (i.e., fluorescence intensity) in this model organism, simulating the immunosuppression and physiological immunodeficiency state after chemotherapy, serves as a direct indicator for efficacy evaluation.

[0065] Example 1: Determination of the maximum tolerated concentration of a single sample

[0066] This embodiment is designed to determine the highest non-lethal concentrations of medium- and long-chain fatty acid triglycerides and nervonic acid that zebrafish can tolerate, i.e., the maximum tolerated concentration (MTC) of a single sample.

[0067] Three-day-old (3 dpf) transgenic T-cell red fluorescent zebrafish were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well (group). A normal control group, a model control group, and a single-sample control group were set up, with a volume of 3 mL per well. Except for the normal control group, all other groups were intravenously injected with vinorelbine tartrate injection to establish a zebrafish immunodeficiency model. The single-sample control groups were administered water-soluble single-samples at concentrations shown in Table 1. After treatment at 28°C for 48 h, the maximum tolerated concentration of the single sample in the model zebrafish was determined.

[0068] Table 1. Results of maximum tolerated concentration for individual samples (n = 30)

[0069] As shown in Table 1, the maximum tolerated concentration (MTC) for MLCT and NA was 2000 μg / mL. Subsequent dosing experiments were designed based on these maximum tolerated concentrations.

[0070] Example 2: Immunotherapy Evaluation Experiment – ​​Efficacy of Increasing T Cell Fluorescence Intensity

[0071] T cells, as the core executors of adaptive immunity, play an irreplaceable role in mediating cellular immunity and assisting humoral immunity. T cells occupy a central position in immune regulation, leading the body's specific immune response, immune memory formation, and tumor immune surveillance; they are the top-level regulatory cells maintaining the overall immune structure. This embodiment primarily examines the effect of the test sample on improving T cell reduction in a zebrafish model of weakened immunity.

[0072] Three-day-old (3 dpf) transgenic T-cell red fluorescent zebrafish were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well (group). Each well contained 3 mL of liquid. Normal control group, model control group, positive control group, single-sample control group, and experimental group were established. Except for the normal control group, all other control groups and experimental groups received intravenous injection of 0.2 mg / mL vinorelbine tartrate injection to establish a zebrafish immunodeficiency model, with an injection volume of 10 nL / zebrafish.

[0073] For the zebrafish in the positive control group, Bailin capsules were administered in water at a dose of 15.0 μg / mL; for the single-sample control group, different concentrations of MLCT or NA were administered in water; for the experimental group, different combinations of MLCT and NA were administered in water. The composition of the samples given to the single-sample control group and the experimental group is shown in Table 2.

[0074] After treatment at 28℃ for 48 h, all experimental and control groups underwent treatment. Ten zebrafish were randomly selected from each group and photographed under a fluorescence microscope. Data were acquired using NIS-Elements D 3.20 advanced image processing software. Because T cells are mainly concentrated in the thymus tissue, and their dense aggregation and overlapping make single-cell counting difficult, semi-quantitative analysis was performed using the fluorescence intensity of T cells in the thymus. The statistical analysis results of this index were used to evaluate the efficacy of the samples in increasing T cell fluorescence intensity. Higher fluorescence intensity indicates higher immunity; therefore, the statistical analysis results of T cell fluorescence intensity can be used to evaluate the immune-enhancing efficacy of the samples.

[0075] Statistical results are expressed as mean ± standard deviation (mean ± SE). Statistical analysis was performed using SPSS software, and a p-value < 0.05 was considered statistically significant.

[0076] In Table 2 below, the synergistic increase in T cell fluorescence intensity in the experimental group was calculated using the following formula (a-1):

[0077]

[0078] Equation (a-1)

[0079] The cooperation coefficient for the increase in T cell fluorescence intensity in the experimental group can be calculated according to the following formula (a-2):

[0080]

[0081] Equation (a-2)

[0082] The unit dose cooperativity coefficient for the increase in T cell fluorescence intensity in the experimental group can be calculated according to the following formula (a-3):

[0083]

[0084] Equation (a-3).

[0085] Table 2. Evaluation results of the efficacy of the samples in improving T-cell reduction (n = 30)

[0086]

[0087] From Table 2 and Figure 1 The results showed that the T cell fluorescence intensity in the model control group was significantly reduced compared with the normal control group, indicating that the zebrafish immunodeficiency model was successfully established by intravenous injection of vinorelbine tartrate injection.

[0088] Compared with the model control group, the T cell fluorescence intensity of the positive control group given Bailin capsules was increased, indicating that the reduction of T cells in the zebrafish model was improved, and thus its immunity was improved.

[0089] Compared with the model control group, the T cell fluorescence intensity of the single-sample control group that was given MLCT (1500 μg / mL) alone was increased, indicating that MLCT alone can improve immunity.

[0090] Compared with the model control group, the experimental groups using the combination of MLCT and NA showed a significant increase in T cell fluorescence intensity (P<0.05). For example, the experimental groups given formulations 4 to 6 all showed a significant increase in T cell fluorescence intensity (P<0.05).

[0091] Furthermore, compared to the single-sample control group, the combination of MLCT and NA showed a significant synergistic effect. For example, as shown in Table 2, formulation 1 (375 μg / mL MLCT + 0.625 μg / mL NA), control formulation 1 (375 μg / mL MLCT), and control formulation 4 (0.625 μg / mL NA) increased the fluorescence intensity of T cells in the zebrafish model by 5105 pixels, 1032 pixels, and 1537 pixels, respectively. The increase in T cell fluorescence intensity caused by formulation 1 (5105 pixels) was much larger than the simple sum of the increases caused by control formulations 1 and 4 (2569 pixels), showing a significant synergistic effect. Similarly, the increases in T cell fluorescence intensity caused by formulations 2 to 6 were also higher than the simple sum of the increases caused by the corresponding amounts of the single sample, i.e., they also showed a significant synergistic effect. Therefore, it can be seen that when MLCT and NA are used in combination, especially at a ratio of 1.5:1 to 2400:1, they have a synergistic effect in improving the fluorescence intensity of T cells in immunocompromised individuals. In other words, the combination of the two can synergistically improve immunity.

[0092] On the other hand, synergistic effects can also be characterized by the synergistic coefficient, which is the ratio of the recovery brought by the formulation containing the combination of MLCT and NA to the sum of the recovery brought by the corresponding two individual samples (comparative formulation). A synergistic coefficient greater than 1 indicates a synergistic effect.

[0093] Furthermore, considering the dosage of the components, the strength of the synergistic effect can be further characterized by the unit dose synergistic coefficient (also known as the contribution rate of the unit dose to the synergistic coefficient). The unit dose synergistic coefficient can reflect the strength of the synergistic effect of formulations containing different doses of MLCT and NA. The larger the unit dose synergistic coefficient, the stronger the contribution of the unit dose to the synergistic effect.

[0094] Since each well has the same volume, the dose is directly proportional to the concentration. For the sake of simplicity, only the concentration is used to calculate the unit dose synergistic coefficient without further incorporating the well volume.

[0095] For example, as shown in Table 2, the recovery of zebrafish T cell fluorescence intensity by formulation 1, control formulation 1, and control formulation 4 compared to the model control group was 5105 pixels, 1032 pixels, and 1537 pixels, respectively. The synergy coefficient of formulation 1 = 5105 / (1032+1537) = 1.99 > 1, indicating that the recovery of zebrafish T cell fluorescence intensity by formulation 1 compared to the model control group (5105 pixels) is greater than the sum of the recovery of zebrafish T cell fluorescence intensity by individual control formulation 1 (1032 pixels) and individual control formulation 4 (1537 pixels) compared to the model control group (2569 pixels). Furthermore, considering the concentrations of each component (MLCT: 375 μg / mL, NA: 0.625 μg / mL), the unit dose synergy coefficient of formulation 1 = 1.99 / (375+0.625) = 5.29 × 10⁻⁶. -3 .

[0096] As shown in Table 2, the synergistic coefficients of formulations 1 to 6 are all greater than 1, indicating that the combination of MLCT and NA has a significant synergistic effect in improving the reduced fluorescence intensity of T cells in immunocompromised individuals. For example, the mass ratio of MLCT to NA in formulations 1 to 6 is 1.5:1 to 2400:1, indicating that when MLCT and NA are used in combination at a mass ratio of 1.5:1 to 2400:1, they have a synergistic effect in improving the reduced fluorescence intensity of T cells in immunocompromised individuals.

[0097] Furthermore, as shown in Table 2, formulations 1 to 6 all exhibit satisfactory unit dose synergy coefficients. In particular, the unit dose synergy coefficients of formulations 1 to 6 are ranked as follows: formulation 4 > formulation 1 > formulation 5 > formulation 3 > formulation 2 > formulation 6, indicating that satisfactory unit dose synergy coefficients can be obtained when the mass ratio of MLCT to NA is within the range of 1.5:1 to 2400:1, preferably 1.5:1 to 1200:1, and more preferably 3:1 to 1200:1.

[0098] Example 3: Immunotherapy Evaluation Experiment – ​​Efficacy of Increasing Neutrophil Fluorescence Intensity

[0099] Neutrophils, as the primary effector cells of innate immunity, are the body's first line of defense against invading pathogens such as bacteria and fungi. They can be rapidly recruited to sites of inflammation and injury, and promptly eliminate invading pathogens through chemotaxis, phagocytosis, and the release of inflammatory mediators and antimicrobial peptides, playing a rapid defensive role in acute anti-infective immune responses. During the body's systemic immune response, T cells, macrophages, and neutrophils, through precise functional coordination, signal transduction, and effector synergy, construct a complete defense network that closely links innate and adaptive immunity, jointly maintaining immune homeostasis, resisting pathogen invasion, and eliminating abnormal cells.

[0100] This embodiment mainly examines the effect of the test sample on improving neutropenia in a zebrafish model of low immunity.

[0101] Transgenic neutrophil-producing green fluorescent zebrafish, 3 days post-fertilization (3 dpf), were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well (group). Each well contained 3 mL of liquid. Normal control group, model control group, positive control group, single-sample control group, and experimental group were established. Except for the normal control group, all other control groups and experimental groups received intravenous injection of 0.2 mg / mL vinorelbine tartrate injection to establish a zebrafish immunodeficiency model, with an injection volume of 10 nL / zebrafish.

[0102] For the zebrafish in the positive control group, Bailin capsules were administered in water at a dose of 15.0 μg / mL; for the single-sample control group, different concentrations of MLCT or NA were administered in water; for the experimental group, different combinations of MLCT and NA were administered in water. The composition of the samples given to the single-sample control group and the experimental group is shown in Table 3.

[0103] After treatment at 28℃ for 48 h, all experimental and control groups had 10 zebrafish randomly selected from each group and photographed under a fluorescence microscope. Data were acquired using NIS-Elements D 3.20 advanced image processing software. Because zebrafish neutrophils are distributed in a discrete, punctate pattern with clear cell boundaries and no significant overlap, cell number, expressed as fluorescence intensity, was used for evaluation. Higher neutrophil fluorescence intensity indicates a greater number of neutrophils, and consequently, higher immunity. Therefore, statistical analysis of neutrophil fluorescence intensity was used to evaluate the immune-enhancing efficacy of the samples. Higher fluorescence intensity correlates with higher immunity; therefore, statistical analysis of neutrophil fluorescence intensity can be used to evaluate the immune-enhancing efficacy of the samples.

[0104] Statistical results are expressed as mean ± standard deviation (mean ± SE). Statistical analysis was performed using SPSS software, and a p-value < 0.05 was considered statistically significant.

[0105] In Table 3 below, the synergistic increase in fluorescence intensity of neutrophils in the experimental group was calculated using the following formula (b-1):

[0106]

[0107] Equation (b-1)

[0108] The synergistic coefficient for the increase in fluorescence intensity of neutrophils in the experimental group can be calculated according to the following formula (b-2):

[0109]

[0110] Equation (b-2)

[0111] The unit dose synergistic coefficient for the increase in fluorescence intensity of neutrophils in the experimental group can be calculated according to the following formula (2-2):

[0112]

[0113] Equation (b-3).

[0114] Table 3. Evaluation results of the efficacy of the samples in improving neutropenia (n = 30)

[0115]

[0116] From Table 3 and Figure 2 The results show that, compared with the normal control group, the fluorescence intensity of neutrophils in the model control group was significantly reduced, indicating that the zebrafish immunodeficiency model was successfully established by intravenous injection of vinorelbine tartrate injection.

[0117] Compared with the model control group, the neutrophil fluorescence intensity of the positive control group given Bailin capsules was increased, indicating that the reduction of neutrophils in the zebrafish model was improved, which in turn indicates that its immunity was improved.

[0118] Compared with the model control group, the neutrophil fluorescence intensity of the single-sample control group that was given MLCT (375 μg / mL, 750 μg / mL or 1500 μg / mL) or NA (250 μg / mL) alone was also increased, indicating that MLCT alone and NA alone can improve immunity.

[0119] Compared with the model control group, the neutrophil fluorescence intensity in the experimental groups using the combination of MLCT and NA was significantly increased (P<0.05). For example, the experimental groups given formulations 1 to 6 all showed a significant increase in neutrophil fluorescence intensity (P<0.05).

[0120] Furthermore, compared to the single-sample control group, the combination of MLCT and NA showed a significant synergistic effect. For example, as shown in Table 3, formulation 1 (375 μg / mL MLCT + 0.625 μg / mL NA), control formulation 1 (375 μg / mL MLCT), and control formulation 4 (0.625 μg / mL NA) increased the fluorescence intensity of neutrophils in the zebrafish model by 4.4 pixels, 3.7 pixels, and 0.4 pixels, respectively. The increase in neutrophil fluorescence intensity caused by formulation 1 (4.4 pixels) was greater than the simple sum of the increases caused by control formulations 1 and 4 (4.1 pixels), demonstrating a significant synergistic effect. Similarly, the increases in neutrophil fluorescence intensity caused by formulations 2 to 6 were also higher than the simple sum of the increases caused by the corresponding amounts of the single sample, thus also demonstrating a significant synergistic effect. Therefore, it can be seen that the combination of MLCT and NA, especially at a ratio of 1.5:1 to 2400:1, has a synergistic effect in improving the fluorescence intensity of neutrophils in immunocompromised individuals, that is, the combination of the two can synergistically improve immunity.

[0121] On the other hand, synergistic effects can also be characterized by the synergistic coefficient, which is the ratio of the recovery brought by the formulation containing the combination of MLCT and NA to the sum of the recovery brought by the corresponding two individual samples (comparative formulation). A synergistic coefficient greater than 1 indicates a synergistic effect.

[0122] Furthermore, considering the dosage of the components, the strength of the synergistic effect can be further characterized by the unit dose synergistic coefficient (also known as the contribution rate of the unit dose to the synergistic coefficient). The unit dose synergistic coefficient can reflect the strength of the synergistic effect of formulations containing different doses of MLCT and NA. The larger the unit dose synergistic coefficient, the stronger the contribution of the unit dose to the synergistic effect.

[0123] Since each well has the same volume, the dose is directly proportional to the concentration. For the sake of simplicity, only the concentration is used to calculate the unit dose synergistic coefficient without further incorporating the well volume.

[0124] For example, as shown in Table 3, Formula 1, Comparative Formula 1, and Comparative Formula 4 resulted in a recovery of neutrophil fluorescence intensity in zebrafish by 4.4 pixels, 3.7 pixels, and 0.4 pixels, respectively, compared to the model control group. The synergy coefficient of Formula 1 = 4.4 / (3.7+0.4) = 1.07 > 1, indicating that the recovery of neutrophil fluorescence intensity in zebrafish by Formula 1 compared to the model control group (4.4 pixels) is greater than the sum of the recovery of neutrophil fluorescence intensity in zebrafish by individual Comparative Formula 1 (3.7 pixels) and Individual Comparative Formula 4 (0.4 pixels) compared to the model control group (4.1 pixels). Furthermore, considering the concentrations of each component (MLCT: 375 μg / mL, NA: 0.625 μg / mL), the unit dose synergy coefficient of Formula 1 = 1.07 / (375+0.625) = 2.86 × 10⁻⁶. -3 .

[0125] As shown in Table 3, the synergistic coefficients of formulations 1 to 6 are all greater than 1, indicating that the combination of MLCT and NA has a significant synergistic effect in improving the reduced fluorescence intensity of neutrophils in immunocompromised individuals. For example, the mass ratio of MLCT to NA in formulations 1 to 6 is 1.5:1 to 2400:1, indicating that when MLCT and NA are used in combination at a mass ratio of 1.5:1 to 2400:1, they have a synergistic effect in improving the reduced fluorescence intensity of neutrophils in immunocompromised individuals.

[0126] Furthermore, as shown in Table 3, formulations 1 to 6 all exhibit satisfactory unit dose synergy coefficients. In particular, the unit dose synergy coefficients of formulations 1 to 6 are ranked as follows: formulation 1 > formulation 3 > formulation 2 > formulation 4 > formulation 5 > formulation 6, indicating that a satisfactory unit dose synergy coefficient can be obtained when the mass ratio of MLCT to NA is within the range of 1.5:1 to 2400:1, preferably 1.5:1 to 1200:1, and more preferably 3:1 to 1200:1.

[0127] Example 4: Immunotherapy Evaluation Experiment – ​​Efficacy of Increasing Macrophage Fluorescence Intensity

[0128] Besides T cells and neutrophils, macrophages also play an important role in the immune system. As innate immune cells with powerful phagocytic functions, macrophages are not only responsible for directly engulfing and clearing pathogens, but also for bridging innate and adaptive immunity and precisely regulating inflammatory responses by presenting antigens, secreting cytokines, and activating T lymphocytes.

[0129] This embodiment mainly examines the effect of the test sample on improving the reduction of macrophages in a zebrafish model of low immunity.

[0130] Three days post-fertilization (3 dpf) transgenic macrophage-derived green fluorescent zebrafish were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well (group). Each well contained 3 mL of liquid. Normal control group, model control group, positive control group, single-sample control group, and experimental group were established. Except for the normal control group, all other control groups and experimental groups received intravenous injection of 0.2 mg / mL vinorelbine tartrate injection to establish a zebrafish immunodeficiency model, with an injection volume of 10 nL / zebrafish.

[0131] For the zebrafish in the positive control group, Bailin capsules were administered in water at a dose of 15.0 μg / mL; for the single-sample control group, different concentrations of MLCT or NA were administered in water; for the experimental group, different combinations of MLCT and NA were administered in water. The composition of the samples given to the single-sample control group and the experimental group is shown in Table 4.

[0132] After treatment at 28℃ for 48 h, all experimental and control groups underwent treatment. Ten zebrafish from each group were randomly selected and photographed under a fluorescence microscope. Data were acquired using NIS-Elements D 3.20 advanced image processing software. Because zebrafish macrophages are both scattered and aggregated at sites of inflammation, the fluorescence intensity of macrophages in the tail vein after the cloaca was used for semi-quantitative analysis. Higher fluorescence intensity indicates higher immunity; therefore, statistical analysis of macrophage fluorescence intensity can be used to evaluate the immune-enhancing efficacy of the samples.

[0133] Statistical results are expressed as mean ± standard deviation (mean ± SE). Statistical analysis was performed using SPSS software, and a p-value < 0.05 was considered statistically significant.

[0134] In Table 4 below, the synergistic increase in fluorescence intensity of macrophages in the experimental group was calculated using the following formula (c-1):

[0135]

[0136] Equation (c-1)

[0137] The synergistic coefficient for the increase in fluorescence intensity in macrophages in the experimental group can be calculated according to the following formula (c-2):

[0138]

[0139] Equation (c-2)

[0140] The unit dose synergistic coefficient for the increase in fluorescence intensity of macrophages in the experimental group can be calculated according to the following formula (c-3):

[0141]

[0142] Equation (c-3).

[0143] Table 4. Evaluation results of the efficacy of the samples in improving macrophage reduction (n = 30)

[0144]

[0145] Compared with the normal control group, the fluorescence intensity of macrophages in the model control group was significantly reduced, indicating that the zebrafish immunodeficiency model was successfully established by intravenous injection of vinorelbine tartrate injection.

[0146] Compared with the model control group, the fluorescence intensity of macrophages in the positive control group given Bailin capsules was increased, indicating that the reduction of macrophages in the zebrafish model was improved, which in turn indicates that its immunity was improved.

[0147] Compared with the model control group, the macrophage fluorescence intensity of the single-sample control group that was given MLCT (750 μg / mL or 1500 μg / mL) or NA (250 μg / mL) alone was also increased, indicating that MLCT alone and NA alone can improve immunity.

[0148] Compared with the model control group, the macrophage fluorescence intensity in the experimental groups using the combination of MLCT and NA was significantly increased (P<0.05). For example, the experimental groups given formulations 1 to 6 all showed a significant increase in macrophage fluorescence intensity (P<0.05).

[0149] Furthermore, compared to the single-sample control group, the combination of MLCT and NA showed a significant synergistic effect. For example, as shown in Table 4, formulation 1 (375 μg / mL MLCT + 0.625 μg / mL NA), control formulation 1 (375 μg / mL MLCT), and control formulation 4 (0.625 μg / mL NA) increased the fluorescence intensity of macrophages in the zebrafish model by 3233 pixels, 1780 pixels, and 1116 pixels, respectively. The increase in macrophage fluorescence intensity caused by formulation 1 (3233 pixels) was greater than the simple sum of the increases caused by control formulations 1 and 4 (2896 pixels), demonstrating a significant synergistic effect. Similarly, the increases in macrophage fluorescence intensity caused by formulations 2 to 6 were also higher than the simple sum of the increases caused by the corresponding amounts of the single sample, thus also showing a significant synergistic effect. Therefore, it can be seen that when MLCT and NA are used in combination, especially at a ratio of 1.5:1 to 2400:1, they have a synergistic effect in improving the fluorescence intensity of macrophages in immunocompromised individuals. In other words, the combination of the two can synergistically improve immunity.

[0150] On the other hand, synergistic effects can also be characterized by the synergistic coefficient, which is the ratio of the recovery brought by the formulation containing the combination of MLCT and NA to the sum of the recovery brought by the corresponding two individual samples (comparative formulation). A synergistic coefficient greater than 1 indicates a synergistic effect.

[0151] Furthermore, considering the dosage of the components, the strength of the synergistic effect can be further characterized by the unit dose synergistic coefficient (also known as the contribution rate of the unit dose to the synergistic coefficient). The unit dose synergistic coefficient can reflect the strength of the synergistic effect of formulations containing different doses of MLCT and NA. The larger the unit dose synergistic coefficient, the stronger the contribution of the unit dose to the synergistic effect.

[0152] Since each well has the same volume, the dose is directly proportional to the concentration. For the sake of simplicity, only the concentration is used to calculate the unit dose synergistic coefficient without further incorporating the well volume.

[0153] For example, as shown in Table 4, the recovery of zebrafish macrophage fluorescence intensity by formulation 1, control formulation 1, and control formulation 4 compared to the model control group was 3233 pixels, 1780 pixels, and 1116 pixels, respectively. The synergy coefficient of formulation 1 = 3233 / (1780+1116) = 1.12 > 1, indicating that the recovery of zebrafish macrophage fluorescence intensity by formulation 1 compared to the model control group (3233 pixels) is greater than the sum of the recovery of zebrafish macrophage fluorescence intensity by individual control formulation 1 (1780 pixels) and individual control formulation 4 (1116 pixels) compared to the model control group (2896 pixels). Furthermore, considering the concentrations of each component (MLCT: 375 μg / mL, NA: 0.625 μg / mL), the unit dose synergy coefficient of formulation 1 = 1.12 / (375+0.625) = 2.97 × 10⁻⁶. -3 .

[0154] As shown in Table 4, the synergistic coefficients of formulations 1 to 6 are all greater than 1, indicating that the combination of MLCT and NA has a significant synergistic effect in improving the reduced fluorescence intensity of macrophages in immunocompromised individuals. For example, the mass ratio of MLCT to NA in formulations 1 to 6 is 1.5:1 to 2400:1, indicating that when MLCT and NA are used in combination at a mass ratio of 1.5:1 to 2400:1, they have a synergistic effect in improving the reduced fluorescence intensity of macrophages in immunocompromised individuals.

[0155] Furthermore, as shown in Table 4, formulations 1 to 6 all exhibit satisfactory unit dose synergy coefficients. In particular, the unit dose synergy coefficients of formulations 1 to 6 are ranked as follows: formulation 4 > formulation 1 > formulation 2 > formulation 5 > formulation 3 > formulation 6, indicating that a satisfactory unit dose synergy coefficient can be obtained when the mass ratio of MLCT to NA is within the range of 1.5:1 to 2400:1, preferably 1.5:1 to 1200:1, and more preferably 3:1 to 1200:1.

[0156] In summary, in immunocompromised individuals, the composition of this application synergistically increases the number of T cells, neutrophils, and macrophages, thereby improving immunity. This application also validated the synergistic effect of MLCT and osteopontin in improving systemic immunity in a zebrafish immunocompromised model.

[0157] The above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit it. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. These modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application.

Claims

1. Use of a nutritional composition for non-therapeutic purposes for improving immunity, characterized in that, The nutritional composition comprises medium-long chain fatty acid triglyceride and nervonic acid, and the mass ratio of the medium-long chain fatty acid triglyceride to the nervonic acid is in the range of 0.1:1 to 5000:

1.

2. Use according to claim 1, characterized in that, The use is a non-therapeutic purpose use for enhancing immunity and / or improving immune efficacy.

3. Use according to claim 1, characterized in that, The improving immunity includes enhancing systemic immunity, and / or restoring one or more of macrophages, neutrophils and T cells that are reduced in an immunocompromised individual.

4. Use according to claim 1, characterized in that, The mass ratio of the medium-long chain fatty acid triglyceride to the nervonic acid is in the range of 1:1 to 2500:

1.

5. Use according to claim 1, characterized in that, The mass ratio of the medium-long chain fatty acid triglyceride to the nervonic acid is in the range of 1.5:1 to 2400:

1.

6. Use according to any one of claims 1 to 5, characterized in that, The nutritional composition is used in the form of a food product comprising the nutritional composition.

7. Use according to claim 6, characterized in that, The food product is a health food product that helps to enhance immunity.

8. Use according to claim 6, characterized in that, The food product is a baby food product.

9. Use according to claim 6, characterized in that, The food product is one or more selected from the group consisting of dairy products, candies, beverages, breads and cookies.

10. Use according to claim 6, characterized in that, The food product is a milk powder or a fermented food product.

11. Use according to claim 10, characterized in that, The milk powder is one or more selected from the group consisting of infant formula milk powder, child formula milk powder, adolescent formula milk powder, pregnant woman formula milk powder, postoperative patient formula milk powder and middle-aged and elderly formula milk powder.

12. Use of a nutritional composition in the manufacture of a product, characterized in that, The product is for improving immunity, the nutritional composition comprises medium-long chain fatty acid triglyceride and nervonic acid, and the mass ratio of the medium-long chain fatty acid triglyceride to the nervonic acid is in the range of 0.1:1 to 5000:

1.

13. Use according to claim 12, characterized in that, The mass ratio of the medium-long chain fatty acid triglyceride to the nervonic acid is in the range of 1:1 to 2500:

1.

14. Use according to claim 12, characterized in that, The mass ratio of the medium-long chain fatty acid triglyceride to the nervonic acid is in the range of 1.5:1 to 2400:

1.

15. Use according to claim 12, characterized in that, The product is a pharmaceutical product or a health food product that helps to improve immunity.

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