A composition for improving pulmonary nodules and detoxification, its preparation method and applications

Abstract

The present invention discloses a composition for improving lung nodules and detoxification, its preparation method and application, which relates to the technical field of medicine. The composition comprises the following components in parts by weight: 60-64 parts of dandelion extract, 20-30 parts of β-nicotinamide mononucleotide, 2-20 parts of coenzyme Q10, and 2-10 parts of selenium-enriched yeast. The composition is fermented and transformed by selenium-enriched yeast, which is beneficial to increasing the content of active ingredients, converting some other ingredients into active ingredients, and enhancing the drug effect. The components and raw materials used in the composition of the present invention are cheap and easily available, convenient to use, with small toxic and side effects. The provided preparation process is simple in operation, easy to conduct quality control during the production process, and the prepared composition has stable components.

Description

Technical Field

[0001] The present invention relates to the technical field of health products, and particularly relates to a composition for improving pulmonary nodules and detoxification, a preparation method thereof, and an application thereof. Background Art

[0002] The Chinese Guidelines for the Diagnosis and Treatment of Pulmonary Nodules define pulmonary nodules as round or irregular lesions with a diameter ≤ 3 cm, and the imaging manifestation is a shadow with increased density, which can be single or multiple, and the boundary is clear or unclear. According to the different densities of the nodules, they are divided into solid, part-solid, and ground-glass nodules. Among them, the malignant frequency of part-solid nodules is the highest, followed by ground-glass nodules and solid nodules. Studies have found that benign nodules can be inflammatory pseudotumors, tuberculomas, hamartomas, sclerosing hemangiomas, etc., and malignant nodules are mostly manifestations of early lung cancer. In the initial treatment, according to the different natures of pulmonary nodules, they are divided into three levels of high, medium, and low risk, corresponding to surgical treatment, anti-inflammatory treatment, or regular review. For nodules with a smaller diameter (<5 mm) or multiple nodules in the lungs, which do not have surgical indications or are difficult to operate surgically, the therapeutic effect of Western medicine is limited. The treatment of pulmonary nodules with traditional Chinese medicine can make up for the deficiencies of Western medicine treatment and has obvious advantages in reducing the size of nodules, preventing nodule malignancy, avoiding surgical risks, and improving the quality of life of patients, but the effect is slow. In recent years, more and more patients with pulmonary nodules have been found in physical examinations or outpatient examinations. For those patients who refuse surgery or have no surgical indications, based on the physiological characteristics of the patient's lungs, the combination of Western medicine and syndrome-differentiated traditional Chinese medicine conditioning treatment can, to a certain extent, reduce nodules, prevent malignancy, and relieve symptoms. Summary of the Invention

[0003] In view of the above-mentioned defects of the prior art, the present invention provides a composition for improving pulmonary nodules and detoxification, a preparation method thereof, and an application thereof to solve the problems raised in the above background art.

[0004] To achieve the above object, the present invention provides the following technical solutions:

[0005] A composition for improving pulmonary nodules and detoxification, comprising the following components in parts by weight: 60-64 parts of dandelion extract, 20-30 parts of β-nicotinamide mononucleotide, 2-20 parts of coenzyme Q10, and 2-10 parts of selenium-enriched yeast.

[0006] Preferably, a composition for improving pulmonary nodules and detoxification, comprising the following components in parts by weight: 62 parts of dandelion extract, 25 parts of β-nicotinamide mononucleotide, 12 parts of coenzyme Q10, and 8 parts of selenium-enriched yeast.

[0007] Preferably, for a composition for improving lung nodules and detoxifying, the preparation method of dandelion extract is as follows: Take dry dandelion, wash, crush it, add ethanol with a volume fraction of 70-95%, soak the dandelion and ethanol at a material-liquid ratio of 1:(3-6) g / mL for 50-70 min, filter to obtain a first filtrate and a first residue; Add water to the first residue, soak and extract it at a material-liquid ratio of 1:(6-10) g / mL at a temperature of 90-120 °C for 1-3 h, repeat 2-5 times, filter and combine the filtrates to obtain a second filtrate; Combine the first filtrate and the second filtrate, concentrate and dry to obtain dandelion extract.

[0008] Preferably, the preparation method of β-nicotinamide mononucleotide includes the following steps: Mix a pH 6-7 phosphate buffer solution with nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide and magnesium chloride to obtain a mixed solution; Adjust the pH of the mixed solution to 7.5-8 and the temperature to 30-35 °C, add purine nucleoside phosphorylase with a concentration of 5-10 g / L and react for 20-40 min, filter to obtain filtrate A; Add nicotinamide nucleoside kinase with a concentration of 2-6 g / L to filtrate A and react for 60-180 min, filter to obtain filtrate B; Add T4 polynucleotide kinase with a concentration of 20-30 g / L to filtrate B and react for 30-50 min to obtain β-nicotinamide mononucleotide.

[0009] Preferably, nicotinamide riboside chloride is purchased from Guangdong Mingtong Biotechnology Co., Ltd.; adenosine 3'-monophosphate is purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; nicotinamide is purchased from Yunnan Xili Biotechnology Co., Ltd.;

[0010] Preferably, the phosphate buffer solution, nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide and magnesium chloride are mixed in a mass ratio of (20-25):(1-3):(0.5-2):(1-6):1; The volume ratio of the phosphate buffer solution, purine nucleoside phosphorylase, nicotinamide nucleoside kinase and T4 polynucleotide kinase is (30-45):(0.05-0.10):(0.1-0.5):(0.8-1.5).

[0011] Preferably, purine nucleoside phosphorylase is purchased from Shanghai Qincheng Biotechnology Co., Ltd.; T4 polynucleotide kinase is purchased from Yeasen Biotechnology (Shanghai) Co., Ltd.;

[0012] Preferably, the preparation method of selenium-enriched yeast includes inoculating Candida bombi in a selenium-enriched medium with a pH of 5.2-6.0 at a temperature of 31-34 °C and a dissolved oxygen content of 22-50%, and culturing it at a rotation speed of 250-600 rpm for 48-72 h to obtain selenium-enriched yeast with a selenium content of 0.6-1%.

[0013] Preferably, Candida bombicola (Latin name: Starmerella bombicola) was purchased from Shanghai Yanzun Biotechnology Co., Ltd., with the product number YZ-J7104.

[0014] Preferably, the selenium-enriched medium consists of a solvent and a solute. The solvent is water, and the solute and its concentration in the selenium-enriched medium are respectively 40 - 60 g / L of glucose, 1 - 3 mmol / L of sodium selenate, 30 - 45 g / L of MgSO4·7H2O, 4 - 9 g / L of chitosan with a molecular weight of 8000 - 10000 Da, and 15 - 20 g / L of glutamic acid.

[0015] Preferably, a preparation method of a composition for improving lung nodules and detoxifying includes uniformly mixing dandelion extract, β-nicotinamide mononucleotide, coenzyme Q10, and selenium-enriched yeast by weight, placing them in a fermentation tank for aerobic fermentation, and after the fermentation is completed, the target composition is obtained.

[0016] Preferably, a preparation method of a composition for improving lung nodules and detoxifying, the aerobic fermentation includes the following steps:

[0017] S1. When aerobic fermentation is carried out for 1 - 5 days, control the temperature in the fermentation tank at 23 - 27 °C, the pH value at 4.2 - 4.5, and the density of selenium-enriched yeast reaches 1×10 6 CFU / mL or more;

[0018] S2. When aerobic fermentation is carried out for 6 - 10 days, control the temperature in the fermentation tank at 30 - 34 °C, the pH value at 4.0 - 4.3, and the density of selenium-enriched yeast reaches 1×10 8 CFU / mL or more;

[0019] S3. When aerobic fermentation is carried out for 11 - 15 days, control the temperature in the fermentation tank at 35 - 38 °C, the pH value at 3.0 - 4.0, and the density of selenium-enriched yeast reaches 1×10 10 CFU / mL or more, which means the fermentation is completed.

[0020] Compared with the prior art, the beneficial effects of the present invention are:

[0021] (1) The composition is fermented and transformed using selenium-enriched yeast, which is beneficial to improving the content of active ingredients, converting some other ingredients into active ingredients, and increasing the medicinal effect. Each component acts together to achieve the therapeutic effects of improving lung nodules and promoting detoxification in the body.

[0022] (2) The component raw materials used in the composition of the present invention are cheap and easy to obtain, convenient to use, have small toxic and side effects, the provided preparation process is simple to operate, easy to control the quality during the production process, and the prepared composition has stable components.

[0023] (3) The sophorolipid produced by Candida bombicola is a glycolipid biosurfactant. Compared with other yeasts, it can absorb more selenium in the culture medium, increase the selenium content in the yeast, and is beneficial to the subsequent fermentation of the composition. At the same time, sophorolipid can increase the internal enzyme activity of dandelion plants, promote the decomposition of dandelion nutrients, and promote the production of active ingredients such as taraxasterol, choline, inulin, and taraxerol, making the composition highly efficient.

[0024] (4) Experimental data show that the compositions prepared in Example 3 and Comparative Examples 1-6 can reduce the contents of TNF-α, IL-10, IL-6, and IL-1β in the serum of rats. In particular, the treatment effect of Group 3 in Example 3 is the best, which can clear the lungs of toxins and improve the swelling of lung nodules. Detailed implementation manners

[0025] The technical solutions of the present invention will be further described in detail below in conjunction with the specific implementation manners.

[0026] Example 1

[0027] A composition for improving lung nodules and detoxifying, comprising the following components in parts by weight: 60 parts of dandelion extract, 20 parts of β-nicotinamide mononucleotide, 2 parts of coenzyme Q10, and 2 parts of selenium-enriched yeast.

[0028] The preparation method of the dandelion extract is as follows: Take dry dandelion, wash and crush it, add ethanol with a volume fraction of 70%, soak the dandelion and ethanol at a solid-liquid ratio of 1:3 g / mL for 50 min, and filter to obtain the first filtrate and the first filter residue; add water to the first filter residue, soak and extract at a solid-liquid ratio of 1:6 g / mL at a temperature of 90 °C for 1 h, repeat 2 times, and filter and combine the filtrates to obtain the second filtrate; combine the first filtrate and the second filtrate, concentrate and dry at a temperature of 70 °C for 60 min to obtain the dandelion extract.

[0029] The preparation method of the β-nicotinamide mononucleotide includes the following steps: Mix pH 6 phosphate buffer with nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide, and magnesium chloride to obtain a mixed solution; adjust the pH of the mixed solution to 7.5, the temperature to 30 °C, add purine nucleoside phosphorylase with a concentration of 5 g / L and react for 20 min, and filter to obtain filtrate A; add nicotinamide nucleoside kinase with a concentration of 2 g / L to filtrate A and react for 60 min, and filter to obtain filtrate B; add T4 polynucleotide kinase with a concentration of 20 g / L to filtrate B and react for 30 min to obtain β-nicotinamide mononucleotide.

[0030] Among them, phosphate buffer, nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide, and magnesium chloride are mixed in a mass ratio of 20:1:0.5:1:1; the volume ratio of phosphate buffer, purine nucleoside phosphorylase, nicotinamide riboside kinase, and T4 polynucleotide kinase is 30:0.05:0.1:0.8.

[0031] The preparation method of the selenium-enriched yeast includes inoculating Candida bombi in a selenium-enriched medium with a pH of 5.2 at a temperature of 31 °C and a dissolved oxygen content of 22%, and culturing at a rotation speed of 250 rpm for 48 h to obtain selenium-enriched yeast with a selenium content of 0.6%.

[0032] The selenium-enriched medium consists of a solvent and a solute. The solvent is water, and the solute and its concentration in the selenium-enriched medium are 40 g / L of glucose, 1 mmol / L of sodium selenate, 30 g / L of MgSO4·7H2O, 4 g / L of chitosan with a molecular weight of 8000 - 10000 Da, and 15 g / L of glutamic acid.

[0033] A preparation method of a composition for improving lung nodules and detoxification includes uniformly mixing dandelion extract, β-nicotinamide mononucleotide, coenzyme Q10, and selenium-enriched yeast by weight, placing them in a fermentation tank for aerobic fermentation, and obtaining the target composition after fermentation is completed; the aerobic fermentation includes the following steps:

[0034] S1. At 1 day of aerobic fermentation, control the temperature in the fermentation tank at 23 °C, the pH value at 4.2, and the density of selenium-enriched yeast to reach 1×10 6 CFU / mL or more;

[0035] S2. At 6 days of aerobic fermentation, control the temperature in the fermentation tank at 30 °C, the pH value at 4.0, and the density of selenium-enriched yeast to reach 1 CFU / mL or more;

[0036] S3. At 11 days of aerobic fermentation, control the temperature in the fermentation tank at 35 °C, the pH value at 3.0, and the density of selenium-enriched yeast to reach 1×10 10 CFU / mL or more, which means the fermentation is completed;

[0037] Example 2

[0038] A composition for improving lung nodules and detoxification includes the following components by weight: 64 parts of dandelion extract, 30 parts of β-nicotinamide mononucleotide, 20 parts of coenzyme Q10, and 10 parts of selenium-enriched yeast.

[0039] The preparation method of dandelion extract is as follows: Take dry dandelion, wash, crush it, add ethanol with a volume fraction of 95%, soak the dandelion and ethanol at a solid-liquid ratio of 1:6 g / mL for 70 min, filter to obtain the first filtrate and the first filter residue; Add water to the first filter residue, soak and extract at a solid-liquid ratio of 1:10 g / mL at a temperature of 120 °C for 3 h, repeat 5 times, filter and combine the filtrates to obtain the second filtrate; Combine the first filtrate and the second filtrate, concentrate and dry at a temperature of 70 °C for 60 min to obtain dandelion extract.

[0040] The preparation method of the β-nicotinamide mononucleotide comprises the following steps: Mix a pH 7 phosphate buffer solution with nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide and magnesium chloride to obtain a mixed solution; Adjust the pH of the mixed solution to 8, the temperature is 35 °C, add purine nucleoside phosphorylase with a concentration of 10 g / L and react for 40 min, filter to obtain filtrate A; Add nicotinamide nucleoside kinase with a concentration of 6 g / L to filtrate A and react for 180 min, filter to obtain filtrate B; Add T4 polynucleotide kinase with a concentration of 30 g / L to filtrate B and react for 50 min to obtain β-nicotinamide mononucleotide.

[0041] The phosphate buffer solution, nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide and magnesium chloride are mixed in a mass ratio of 25:3:2:6:1; The volume ratio of the phosphate buffer solution, purine nucleoside phosphorylase, nicotinamide nucleoside kinase and T4 polynucleotide kinase is 45:0.10:0.5:1.5.

[0042] The preparation method of the selenium-enriched yeast comprises inoculating Candida bombi in a selenium-enriched medium with a pH of 6.0 at a temperature of 34 °C and a dissolved oxygen content of 50%, and culturing at a rotation speed of 600 rpm for 72 h to obtain selenium-enriched yeast with a selenium content of 1%.

[0043] The selenium-enriched medium consists of a solvent and a solute. The solvent is water, and the solute and its concentration in the selenium-enriched medium are respectively 60 g / L of glucose, 3 mmol / L of sodium selenate, 45 g / L of MgSO4·7H2O, 9 g / L of chitosan with a molecular weight of 8000 - 10000 Da and 20 g / L of glutamic acid.

[0044] A preparation method of a composition for improving lung nodules and detoxifying, comprising mixing dandelion extract, β-nicotinamide mononucleotide, coenzyme Q10 and selenium-enriched yeast evenly by weight, placing them in a fermentation tank for aerobic fermentation, and obtaining the target composition after the fermentation is completed; The aerobic fermentation comprises the following steps:

[0045] S1. When aerobic fermentation is carried out for 5 d, control the temperature in the fermentation tank at 27 °C, the pH value at 4.5, and the density of selenium-enriched yeast reaches 1×10 6 CFU / mL or more;

[0046] S2. At 10 days of aerobic fermentation, control the temperature in the fermenter at 34 °C, the pH value at 4.3, and the density of selenium-enriched yeast reaches 10 8 CFU / mL or more;

[0047] S3. At 15 days of aerobic fermentation, control the temperature in the fermenter at 38 °C, the pH value at 4.0, and the density of selenium-enriched yeast reaches 1×10 10 CFU / mL or more, which means the fermentation is completed.

[0048] Example 3

[0049] A composition for improving lung nodules and detoxifying, comprising the following components in parts by weight: 62 parts of dandelion extract, 25 parts of β-nicotinamide mononucleotide, 12 parts of coenzyme Q10, and 8 parts of selenium-enriched yeast.

[0050] The preparation method of the dandelion extract is as follows: Take dry dandelion, wash and crush it, add ethanol with a volume fraction of 90%, soak the dandelion and ethanol at a material-liquid ratio of 1:4 g / mL for 60 min, and filter to obtain the first filtrate and the first filter residue; Add water to the first filter residue, soak and extract at a material-liquid ratio of 1:9 g / mL at a temperature of 100 °C for 2 h, repeat 3 times, and filter and combine the filtrates to obtain the second filtrate; Combine the first filtrate and the second filtrate, concentrate and dry at a temperature of 70 °C for 60 min to obtain the dandelion extract.

[0051] The preparation method of the β-nicotinamide mononucleotide includes the following steps: Mix phosphate buffer solution with a pH of 6.2, nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide, and magnesium chloride to obtain a mixed solution; Adjust the pH of the mixed solution to 7.8, the temperature to 32 °C, add purine nucleoside phosphorylase with a concentration of 8 g / L and react for 30 min, and filter to obtain filtrate A; Add nicotinamide nucleoside kinase with a concentration of 3 g / L to filtrate A and react for 100 min, and filter to obtain filtrate B; Add T4 polynucleotide kinase with a concentration of 25 g / L to filtrate B and react for 40 min to obtain β-nicotinamide mononucleotide.

[0052] The phosphate buffer solution, nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide, and magnesium chloride are mixed in a mass ratio of 22:2:1.5:5:1; The volume ratio of the phosphate buffer solution, purine nucleoside phosphorylase, nicotinamide nucleoside kinase, and T4 polynucleotide kinase is 40:0.08:0.2:1.0.

[0053] The preparation method of the selenium-enriched yeast includes inoculating Candida bombi in a selenium-enriched medium with a pH of 5.5 at a temperature of 32 °C and a dissolved oxygen content of 30%, and culturing at a rotation speed of 400 rpm for 36 h to obtain selenium-enriched yeast with a selenium content of 0.8%.

[0054] The selenium-enriched medium consists of a solvent and a solute. The solvent is water, and the solute and its concentration in the selenium-enriched medium are 50 g / L of glucose, 2 mmol / L of sodium selenate, 40 g / L of MgSO4·7H2O, 6 g / L of chitosan with a molecular weight of 8000 - 10000 Da, and 18 g / L of glutamic acid respectively.

[0055] A preparation method of a composition for improving lung nodules and detoxification includes uniformly mixing dandelion extract, β-nicotinamide mononucleotide, coenzyme Q10, and selenium-enriched yeast by weight, placing them in a fermentation tank for aerobic fermentation, and after the fermentation is completed, the target composition is obtained; the aerobic fermentation includes the following steps:

[0056] S1. At 2 days of aerobic fermentation, control the temperature in the fermentation tank at 25 °C, the pH value at 4.3, and the density of selenium-enriched yeast reaches 1×10 6 CFU / mL or more;

[0057] S2. At 5 days of aerobic fermentation, control the temperature in the fermentation tank at 32 °C, the pH value at 4.2, and the density of selenium-enriched yeast reaches 1×10 8 CFU / mL or more;

[0058] S3. At 10 days of aerobic fermentation, control the temperature in the fermentation tank at 36 °C, the pH value at 3.5, and the density of selenium-enriched yeast reaches 1×10 10 CFU / mL or more, which means the fermentation is completed.

[0059] Comparative Example 1

[0060] The difference between this comparative example and Example 3 is that the dandelion extract is replaced with a prunella vulgaris extract, and the other components and steps remain unchanged.

[0061] A composition for improving lung nodules and detoxification includes the following components by weight: 62 parts of prunella vulgaris extract, 25 parts of β-nicotinamide mononucleotide, 12 parts of coenzyme Q10, and 8 parts of selenium-enriched yeast.

[0062] The preparation method of the prunella vulgaris extract is as follows: Take dried prunella vulgaris, wash and crush it, add ethanol with a volume fraction of 90%, soak the prunella vulgaris and ethanol at a material-liquid ratio of 1:4 g / mL for 60 min, and filter to obtain a first filtrate and a first filter residue; add water to the first filter residue, soak and extract it at a material-liquid ratio of 1:9 g / mL at a temperature of 100 °C for 2 h, repeat 3 times, and filter and combine the filtrates to obtain a second filtrate; combine the first filtrate and the second filtrate, and concentrate and dry them at a temperature of 70 °C for 60 min to obtain the dandelion extract.

[0063] A preparation method of a composition for improving lung nodules and detoxification, comprising mixing Prunella vulgaris extract, β-nicotinamide mononucleotide, coenzyme Q10 and selenium-enriched yeast evenly according to weight parts, placing them in a fermentation tank for aerobic fermentation, and obtaining the target composition after fermentation is completed; the aerobic fermentation includes the following steps:

[0064] S1. At 2 days of aerobic fermentation, control the temperature in the fermentation tank at 25 °C, the pH value at 4.3, and the density of selenium-enriched yeast to reach 1×10 6 CFU / mL or more;

[0065] S2. At 5 days of aerobic fermentation, control the temperature in the fermentation tank at 32 °C, the pH value at 4.2, and the density of selenium-enriched yeast to reach 1×10 8 CFU / mL or more;

[0066] S3. At 10 days of aerobic fermentation, control the temperature in the fermentation tank at 36 °C, the pH value at 3.5, and the density of selenium-enriched yeast to reach 1×10 10 CFU / mL or more, which means the fermentation is completed.

[0067] Comparative Example 2

[0068] The difference between this comparative example and Example 3 is that β-nicotinamide mononucleotide is replaced with nicotinamide, and the other components and steps remain unchanged.

[0069] A composition for improving lung nodules and detoxification, comprising the following components in parts by weight: 62 parts of dandelion extract, 25 parts of nicotinamide, 12 parts of coenzyme Q10, and 8 parts of selenium-enriched yeast.

[0070] A preparation method of a composition for improving lung nodules and detoxification, comprising mixing dandelion extract, nicotinamide, coenzyme Q10 and selenium-enriched yeast evenly according to weight parts, placing them in a fermentation tank for aerobic fermentation, and obtaining the target composition after fermentation is completed; the aerobic fermentation includes the following steps:

[0071] S1. At 2 days of aerobic fermentation, control the temperature in the fermentation tank at 25 °C, the pH value at 4.3, and the density of selenium-enriched yeast to reach 1×10 6 CFU / mL or more;

[0072] S2. At 5 days of aerobic fermentation, control the temperature in the fermentation tank at 32 °C, the pH value at 4.2, and the density of selenium-enriched yeast to reach 1×10 8 CFU / mL or more;

[0073] S3. At 10 days of aerobic fermentation, control the temperature in the fermentation tank at 36 °C, the pH value at 3.5, and the density of selenium-enriched yeast to reach 1×10 10 CFU / mL or more, which means the fermentation is completed.

[0074] Comparative Example 3

[0075] The difference between this comparative example and Example 3 is that the preparation method of β-nicotinamide mononucleotide does not involve segmented enzymatic hydrolysis, and the other components and steps remain unchanged.

[0076] A composition for improving lung nodules and detoxification, comprising the following components in parts by weight: 62 parts of dandelion extract, 25 parts of β-nicotinamide mononucleotide, 12 parts of coenzyme Q10, and 8 parts of selenium-enriched yeast.

[0077] The preparation method of the β-nicotinamide mononucleotide comprises the following steps: at a temperature of 32 °C, reacting a pH 6.2 phosphate buffer solution with nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide, magnesium chloride concentration, 8 g / L purine nucleoside phosphorylase

[0078] , 3 g / L nicotinamide riboside kinase concentration, and 25 g / L T4 polynucleotide kinase for 170 min to obtain β-nicotinamide mononucleotide.

[0079] The phosphate buffer solution, nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide, and magnesium chloride are mixed in a mass ratio of 22:2:1.5:5:1; the volume ratio of the phosphate buffer solution, purine nucleoside phosphorylase, nicotinamide riboside kinase, and T4 polynucleotide kinase is 40:0.08:0.2:1.0.

[0080] A preparation method of a composition for improving lung nodules and detoxification, comprising mixing dandelion extract, β-nicotinamide mononucleotide, coenzyme Q10, and selenium-enriched yeast in parts by weight evenly, placing them in a fermentation tank for aerobic fermentation, and obtaining the target composition after fermentation is completed; the aerobic fermentation comprises the following steps:

[0081] S1. At 2 days of aerobic fermentation, control the temperature in the fermentation tank at 25 °C, the pH value at 4.3, and the density of selenium-enriched yeast to reach 1×10 6 CFU / mL or more;

[0082] S2. At 5 days of aerobic fermentation, control the temperature in the fermentation tank at 32 °C, the pH value at 4.2, and the density of selenium-enriched yeast to reach 1×10 8 CFU / mL or more;

[0083] S3. At 10 days of aerobic fermentation, control the temperature in the fermentation tank at 36 °C, the pH value at 3.5, and the density of selenium-enriched yeast to reach 1×10 10 CFU / mL or more, which means the fermentation is completed.

[0084] Comparative Example 4

[0085] The difference between this comparative example and Example 3 is that the selenium-enriched yeast is replaced with non-selenium-enriched yeast, and the other components and steps remain unchanged.

[0086] A composition for improving lung nodules and detoxification, comprising the following components in parts by weight: 62 parts of dandelion extract, 25 parts of β-nicotinamide mononucleotide, 12 parts of coenzyme Q10, and 8 parts of Candida bombi.

[0087] A preparation method of a composition for improving lung nodules and detoxification, comprising mixing dandelion extract, β-nicotinamide mononucleotide, coenzyme Q10 and Candida bombi evenly according to parts by weight, placing them in a fermentation tank for aerobic fermentation, and obtaining the target composition after fermentation is completed; the aerobic fermentation includes the following steps:

[0088] S1. When aerobic fermentation is carried out for 2 days, control the temperature in the fermentation tank at 25 °C, the pH value at 4.3, and the density of Candida bombi reaches 1×10 6 CFU / mL or more;

[0089] S2. When aerobic fermentation is carried out for 5 days, control the temperature in the fermentation tank at 32 °C, the pH value at 4.2, and the density of Candida bombi reaches 1×10 8 CFU / mL or more;

[0090] S3. When aerobic fermentation is carried out for 10 days, control the temperature in the fermentation tank at 36 °C, the pH value at 3.5, and the density of Candida bombi reaches 1×10 10 CFU / mL or more, which means the fermentation is completed.

[0091] Comparative Example 5

[0092] The difference between this comparative example and Example 3 is that: replacing Candida bombi enriched with selenium with Saccharomyces cerevisiae enriched with selenium, and the other components and steps remain unchanged.

[0093] A composition for improving lung nodules and detoxification, comprising the following components in parts by weight: 62 parts of dandelion extract, 25 parts of β-nicotinamide mononucleotide, 12 parts of coenzyme Q10, and 8 parts of selenium-enriched yeast.

[0094] The preparation method of the selenium-enriched yeast includes inoculating Saccharomyces cerevisiae into a selenium-enriched medium with a pH of 5.5 at a temperature of 32 °C and a dissolved oxygen content of 30%, and culturing it at a rotation speed of 400 rpm for 36 hours to obtain selenium-enriched yeast with a selenium content of 0.8%.

[0095] The selenium-enriched medium is composed of a solvent and a solute. The solvent is water, and the solute and its concentration in the selenium-enriched medium are 50 g / L of glucose, 2 mmol / L of sodium selenate, 40 g / L of MgSO4·7H2O, 6 g / L of chitosan with a molecular weight of 8000-10000 Da, and 18 g / L of glutamic acid.

[0096] A method for preparing a composition for improving lung nodules and detoxifying, comprising uniformly mixing dandelion extract, β-nicotinamide mononucleotide, coenzyme Q10 and selenium-enriched yeast according to weight, placing the mixture in a fermentation tank for aerobic fermentation, and obtaining the target composition after the fermentation is completed; the aerobic fermentation comprises the following steps:

[0097] S1. During the 2nd day of aerobic fermentation, the temperature in the fermentation tank was controlled at 25°C, the pH value was 4.3, and the density of selenium-enriched yeast reached 1×10 6 CFU / mL or above;

[0098] S2. During the 5th day of aerobic fermentation, the temperature in the fermentation tank was controlled at 32°C, the pH value was 4.2, and the density of selenium-enriched yeast reached 1×10 8 CFU / mL or above;

[0099] S3. After 10 days of aerobic fermentation, the temperature in the fermentation tank was controlled at 36°C, the pH value was 3.5, and the density of selenium-enriched yeast reached 1×10 10 When the CFU / mL is above, the fermentation is complete.

[0100] Comparative Example 6

[0101] The difference between this comparative example and Example 3 is that the preparation method of the composition does not adopt segmented fermentation, and the other components and steps remain unchanged.

[0102] A preparation method of a composition for improving lung nodules and detoxification, comprising mixing dandelion extract, β-nicotinamide mononucleotide, coenzyme Q10 and selenium-enriched yeast in parts by weight, placing the mixture in a fermenter for aerobic fermentation for 17 days, controlling the temperature in the fermenter at 36°C, the pH value at 3.5, and the density of the selenium-enriched yeast at 1×10 10 CFU / mL or above, the fermentation is complete and the target composition is obtained.

[0103] Experimental Example 1 Test on Inhibition of Pulmonary Nodule Swelling

[0104] 1.1 Experimental Materials

[0105] The target compositions prepared in Examples 1 to 3 and Comparative Examples 1 to 6; 120 SD male rats, weighing 200 to 220 g;

[0106] 1.2 Modeling method

[0107] Each group of rats was intraperitoneally injected with a 3.0% sodium pentobarbital solution (dose of 45 mg / kg body weight) and fixed after anesthesia. The rat neck skin was disinfected with 75.0% ethanol, and the neck was cut along the midline of the neck, and the muscles were bluntly separated to expose the trachea. Non-exposed endotracheal intubation was used for poisoning, and 250 μL of silica suspension with a mass concentration of 300 g / L was injected into the trachea at one time;

[0108] 1.3 Detection indicators

[0109]

[0110] 1.4 Experimental methods

[0111] 120 male SD rats, 10 rats in each group, were divided into the groups of Example 1 - 3, the groups of Comparative Example 1 - 6, the model group, the control group (commercially available product: Guizhi Fuling Capsule, composed of Ramulus Cinnamomi, Poria, Persica, Radix Paeoniae Alba, and Cortex Moutan), and the blank group; among them, the groups of Example 1 - 3 were model rats + the compositions prepared in Example 1 - 3; the groups of Comparative Example 1 - 6 were model rats + the compositions prepared in Comparative Example 1 - 6; the model group was model rats + an equal amount of normal saline; the control group was model rats + an equal amount of commercially available product; the blank group was normal rats + an equal amount of normal saline; the administration dose was 10 g / kg / day for a total of 45 days. After the administration ended, the rats in each group were anesthetized and sacrificed, the weight of the lung tissue of the rats was weighed, and the lung swelling coefficient and the inhibition rate of lung nodule swelling were calculated;

[0112] 1.5 Experimental results

[0113] The results are shown in Table 1:

[0114] Table 1 Swelling test

[0115]

[0116]

[0117] As can be seen from Table 1, the compositions of Example 1 - 3 prepared by the present invention have the best inhibition rate of lung nodule swelling and the best effect. By comparing Example 3 with Comparative Example 1, Comparative Example 2, and Comparative Example 5, it can be known that if the dandelion extract is replaced with Prunella vulgaris water extract, or β - nicotinamide mononucleotide is replaced with nicotinamide, or selenium - rich Bombus terrestris Candida is replaced with selenium - rich baker's yeast, the expected effect cannot be achieved, indicating that the technical solution of the present invention has selectivity for components, and the selection of yeast affects the effectiveness of the composition; by comparing Example 3 with Comparative Example 3 and 4, it can be known that the active ingredients in β - nicotinamide mononucleotide prepared by non - segmented enzymatic hydrolysis are lower than those prepared by segmented enzymatic hydrolysis, and its inhibition rate of lung nodule swelling will decrease. The rich selenium element in yeast has a great impact on the overall formula effect; by comparing Example 3 with Comparative Example 6, it can be known that segmented fermentation is beneficial to the leaching of active ingredients, and different active ingredients require different leaching conditions, and the present technical solution has a certain selectivity for the preparation method.

[0118] Experimental Example 2 Serum detection

[0119] 2.1 Experimental materials

[0120] The target compositions prepared in Example 3 and Comparative Examples 1-6; 100 male SD rats, weighing 200-220 g;

[0121] 2.2 Modeling method

[0122] Each group of rats was intraperitoneally injected with a 3.0% sodium pentobarbital solution (dose: 45 mg / kg body weight). After anesthesia, they were fixed. The neck skin of the rats was disinfected with 75.0% ethanol by volume. An incision was made along the midline of the neck, and the muscles were bluntly separated to expose the trachea. Non-exposed tracheal intubation was used for exposure to the toxin. A single injection of 250 μL of a 300 g / L silicon dioxide suspension was injected into the trachea;

[0123] 2.3 Detection indicators

[0124] Serum TNF-α (tumor necrosis factor), IL-10 (interleukin 10), IL-6 (interleukin 6) and IL-1β (interleukin 1β);

[0125] 2.4 Experimental method

[0126] 100 male SD rats, 10 in each group, were divided into Example 1 group, Comparative Examples 1-6 groups, model group and control group (commercially available product: Guizhi Fuling Capsule, composed of cinnamon twig, poria cocos, peach kernel, white peony root, and moutan bark) and blank group; among them, the Example 1 group was model rats + the composition prepared in Examples 1-3; the Comparative Examples 1-6 groups were model rats + the composition prepared in Comparative Examples 1-6; the model group was model rats + an equal amount of normal saline; the control group was model rats + an equal amount of commercially available product; the blank group was normal rats + an equal amount of normal saline; the administration dose was 10 g / kg / day for a total of 45 days. After the administration ended, the rats in each group were anesthetized and sacrificed, and blood was taken from the abdominal aorta and then sacrificed. After the whole blood was left standing at room temperature for 30 min, it was centrifuged at 3500 r / min for 10 min at 4°C, and the supernatant was taken. The contents of TNF-α, IL-10, IL-6 and IL-1β in the serum were detected by ELISA method;

[0127] 2.5 Experimental results

[0128] As shown in Table 2:

[0129] Table 2 Test results of the contents of TNF-α, IL-10, IL-6 and IL-1β in serum

[0130]

[0131] As can be seen from Table 2, compared with the blank group, the contents of TNF-α, IL-10, IL-6 and IL-1β in the serum of rats in the model group increased; compared with the model group, the contents of TNF-α, IL-10, IL-6 and IL-1β in the serum of rats in Example 3 group, Comparative Example 1-6 groups and the control group decreased, and the various indicators of Example 3 group were closer to the blank group.

[0132] Toxicity experiment of Experimental Example 3

[0133] The acute toxicity test showed that the target composition prepared in Example 3 was continuously intragastrically administered to 30 mice at 30 times, 50 times and 100 times the human dosage for three days, and observed for seven days. All the mice survived healthily without any toxic reaction.

[0134] The above are only the preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A composition for improving lung nodules and detoxification, characterized in that, Comprising the following components in parts by weight: 60 - 64 parts of dandelion extract, 20 - 30 parts of β-nicotinamide mononucleotide, 2 - 20 parts of coenzyme Q10, 2 - 10 parts of selenium-enriched yeast; The preparation method of β-nicotinamide mononucleotide comprises the following steps: Mix phosphate buffer solution with pH 6 - 7, nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide and magnesium chloride to obtain a mixed solution; adjust the pH of the mixed solution to 7.5 - 8, the temperature to 30 - 35 °C, add purine nucleoside phosphorylase with a concentration of 5 - 10 g / L and react for 20 - 40 min, then filter to obtain filtrate A; add nicotinamide riboside kinase with a concentration of 2 - 6 g / L to filtrate A and react for 60 - 180 min, then filter to obtain filtrate B; add T4 polynucleotide kinase with a concentration of 20 - 30 g / L to filtrate B and react for 30 - 50 min to obtain β-nicotinamide mononucleotide; The preparation method of selenium-enriched yeast comprises inoculating Candida bombi in a selenium-enriched medium with pH 5.2 - 6.0 at a temperature of 31 - 34 °C and a dissolved oxygen content of 22 - 50%, and culturing at a rotation speed of 250 - 600 rpm for 48 - 72 h to obtain selenium-enriched yeast with a selenium content of 0.6 - 1%; The preparation method of the composition for improving lung nodules and detoxifying comprises mixing dandelion extract, β-nicotinamide mononucleotide, coenzyme Q10 and selenium-enriched yeast evenly by weight, placing them in a fermentation tank for aerobic fermentation, and obtaining the target composition after fermentation is completed; The aerobic fermentation comprises the following steps: S1. When the aerobic fermentation lasts for 1 - 5 days, control the temperature in the fermenter at 23 - 27 °C, the pH value at 4.2 - 4.5, and the density of selenium-enriched yeast reaches 1×10 6 CFU / mL or more; S2. When the aerobic fermentation lasts for 6 to 10 days, control the temperature in the fermenter at 30 to 34 °C, the pH value at 4.0 to 4.3, and the density of selenium-enriched yeast reaches 1×10 8 CFU / mL or more; S3. When the aerobic fermentation lasts for 11 - 15 days, control the temperature in the fermenter at 35 - 38 °C, the pH value at 3.0 - 4.0, and the density of selenium-enriched yeast reaches 1×10 10 CFU / mL or more, then the fermentation is completed.

2. The composition for improving pulmonary nodules and detoxification according to claim 1, characterized in that, Comprising the following components in parts by weight: 62 parts of dandelion extract, 25 parts of β-nicotinamide mononucleotide, 12 parts of coenzyme Q10, 8 parts of selenium-enriched yeast.

3. The composition for improving pulmonary nodules and detoxification according to claim 1, characterized in that, The preparation method of dandelion extract is as follows: Take dry dandelion, wash, crush and add ethanol with a volume fraction of 70 - 95%, soak the dandelion and ethanol at a material-liquid ratio of 1:(3 - 6) g / mL for 50 - 70 min, filter to obtain the first filtrate and the first filter residue; add water to the first filter residue, soak and extract at a material-liquid ratio of 1:(6 - 10) g / mL at a temperature of 90 - 120 °C for 1 - 3 h, repeat 2 - 5 times, filter and combine the filtrates to obtain the second filtrate; combine the first filtrate and the second filtrate, concentrate and dry to obtain dandelion extract.

4. A composition for improving pulmonary nodules and detoxification according to claim 1, characterized in that, Phosphate buffer solution, nicotinamide riboside chloride, adenosine 3'-monophosphate, nicotinamide and magnesium chloride are mixed in a mass ratio of (20 - 25):(1 - 3):(0.5 - 2):(1 - 6):1; the volume ratio of phosphate buffer solution, purine nucleoside phosphorylase, nicotinamide riboside kinase and T4 polynucleotide kinase is (30 - 45):(0.05 - 0.10):(0.1 - 0.5):(0.8 - 1.5).

5. A composition for improving pulmonary nodules and detoxification according to claim 1, characterized in that, The selenium-enriched medium consists of a solvent and a solute, the solvent is water, and the solute and its concentration in the selenium-enriched medium are respectively 40 - 60 g / L of glucose, 1 - 3 mmol / L of sodium selenate, 30 - 45 g / L of MgSO4﹒7H2O, 4 - 9 g / L of chitosan with a molecular weight of 8000 - 10000 Da and 15 - 20 g / L of glutamic acid.

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