A ginseng fermented liquid of Bifidobacterium animalis subspecies lactis ZKBb1223.1 and its application in preparing a drug for improving the appetite of patients undergoing radiotherapy and chemotherapy for tumors

Abstract

The present invention relates to the technical field of probiotic fermentation, and in particular to a ginseng fermentation liquid of Bifidobacterium animalis subspecies ZKBb1223.1 and its application in the preparation of a drug for improving the appetite of patients undergoing radiotherapy and chemotherapy for tumors. Bifidobacterium animalis subsp. lactis )ZKBb1223.1 has been deposited in the General Microbiology Center of China National Committee for the Preservation of Microorganisms on May 13, 2024, with the deposit address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 30550. The fermentation effect of animal Bifidobacterium lactis subspecies ZKBb1223.1 provided by the present invention is good. Clinical trials have shown that the use of animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid can improve the symptoms of anorexia after tumor and related treatments, increase the weight of patients, and improve the quality of life of patients without adverse reactions.

Description

Technical Field

[0001] The invention relates to the technical field of probiotic fermentation, and in particular to a ginseng fermentation liquid of animal Bifidobacterium lactis subspecies ZKBb1223.1 and application thereof in preparing a medicine for improving the appetite of patients undergoing radiotherapy and chemotherapy for tumors. Background Art

[0002] Radiotherapy, chemotherapy and other treatments can cause some adverse reactions in cancer patients, especially gastrointestinal reactions associated with intestinal microecological imbalance, such as nausea, vomiting, loss of appetite, etc., accompanied by weight loss, decreased physical strength and other complications. Data show that about 26%-58% of patients with advanced cancer will experience loss of appetite, and 33% of patients will experience loss of appetite, which affects the quality of life of patients and their confidence in continuing to receive anti-tumor chemotherapy, resulting in reduced tolerance to treatment.

[0003] Clinically, appetite stimulants, such as steroids and progesterone, are most commonly used to improve the symptoms of anorexia. However, such drugs can also cause side effects such as allergies, insomnia, and nausea. In recent years, studies have shown that Chinese medicine has achieved satisfactory clinical efficacy in improving the appetite of patients undergoing radiotherapy and chemotherapy, with few side effects. According to the theory of traditional Chinese medicine, the etiology and pathogenesis of digestive tract reactions after radiotherapy and chemotherapy are mainly spleen and stomach damage and dampness stagnation in the middle jiao. Spleen deficiency leads to inability to transport and transform, stomach disharmony and descent leads to dysfunction of reception and loss of appetite, and dampness and turbidity in the middle lead to abdominal distension, nausea and vomiting. The treatment principle is mainly to strengthen the spleen, eliminate dampness and harmonize the stomach. Ginseng is the dried root and rhizome of ginseng, a plant of the genus Panax, Araliaceae. It has a wide range of and significant pharmacological effects and is known as the "king of herbs". Ginseng has the effects of greatly replenishing vital energy, restoring pulse and consolidating deficiency, replenishing spleen and lungs, promoting fluid and nourishing blood, and calming the mind and improving intelligence. Ginseng is mainly used to treat serious illness, long-term illness, blood loss, dehydration resulting in loss of vital energy, fatigue and weak pulse; and spleen deficiency resulting in loss of appetite, fatigue, vomiting and diarrhea.

[0004] Fermentation is a method of preparing traditional Chinese medicine with a history of thousands of years. However, traditional fermented Chinese medicine is fermented under natural conditions using natural bacteria, and the fermentation conditions are uncontrollable. Modern fermentation uses known microorganisms to control fermentation conditions. Through the complex metabolic activities of microorganisms, the structure and content of active ingredients in traditional Chinese medicine are changed, thereby improving the efficacy and medicinal value of traditional Chinese medicine. Its application field is very wide.

[0005] CN 118058468 A provides a probiotic solid beverage, comprising Lactobacillus plantarum HF05, fermented Lactobacillus mucilaginosus HF06, fermented Lactobacillus mucilaginosus HF07 and Lactococcus lactis HF08, as well as functional components such as dietary fiber, natural fruit powder and ginseng powder. Although the solid beverage has a certain improvement effect on chronic colitis, its effect mainly depends on a postbiotic combination preparation. Since the ginseng powder has not been fermented, its active ingredients are difficult to be well utilized.

[0006] CN 105145819 A discloses a ginseng yogurt having the effects of relieving physical fatigue and regulating the stomach. Ginseng is used to make the product have the effect of relieving physical fatigue, but the effect of ginseng on improving appetite after fermentation is not studied. Summary of the invention

[0007] In view of the technical problem that there is a lack of research on ginseng fermented liquid in improving appetite in the prior art, the present invention provides a ginseng fermented liquid of Bifidobacterium lactis subsp. ZKBb1223.1 and its application in preparing a medicine for improving appetite in patients undergoing radiotherapy and chemotherapy for tumors.

[0008] In a first aspect, the present invention provides a strain of Bifidobacterium animalis subsp. lactis, which is Bifidobacterium animalis subsp. lactis ( Bifidobacterium animalis subsp. lactis ) ZKBb1223.1 was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on May 13, 2024. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No.30550.

[0009] In a second aspect, the present invention provides a ginseng fermented liquid of Bifidobacterium lactis subspecies ZKBb1223.1 capable of improving the appetite of patients with tumor radiotherapy and chemotherapy, which is prepared by the following method:

[0010] (1) Soaking ginseng in water and then boiling it to obtain ginseng extract;

[0011] (2) Add yeast extract powder and oligofructose to the ginseng extract and sterilize for later use;

[0012] (3) Inoculate the sterilized ginseng extract of step (2) with the above-mentioned Bifidobacterium animalis subspecies lactis ZKBb1223.1, shake well, and then culture anaerobically at 37° C. to obtain the product.

[0013] Furthermore, in step (1), the ginseng is soaked in water of eight times its mass for 60 minutes.

[0014] Furthermore, in step (1), the boiling condition is to boil the mixture and then keep boiling for 90 minutes.

[0015] Furthermore, in step (1), after decoction, the filtrate is filtered and separated, and the ginseng extract with a crude drug content of 1.0 g / mL is obtained by decoction and concentration.

[0016] Furthermore, in step (2), based on the volume of the ginseng extract, the amount of yeast extract powder added is 2-4 g / 100 mL, and the amount of fructooligosaccharide added is 5-7 g / 100 mL.

[0017] Furthermore, in step (3), 2 wt% to 4 wt% of a bacterial suspension of Bifidobacterium animalis subspecies lactis ZKBb1223.1 is inoculated into the ginseng extract, and the bacterial suspension is prepared by the following method:

[0018] Animal Bifidobacterium lactis subsp. ZKBb1223.1 was inoculated into MRS liquid culture medium supplemented with 0.05 g / L L-cysteine ​​hydrochloride, cultured anaerobically at 37°C for 24 h, activated and propagated to the 2nd to 3rd generation, and prepared into a bacterial suspension.

[0019] Furthermore, the pH of the ginseng fermentation liquid of Bifidobacterium animalis subspecies lactis ZKBb1223.1 is 3.5-3.9.

[0020] In a third aspect, the present invention also provides a use of the above-mentioned animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid in the preparation of a drug for improving the appetite of patients with tumor radiotherapy and chemotherapy.

[0021] The beneficial effects of the present invention are:

[0022] The animal Bifidobacterium lactis subspecies ZKBb1223.1 provided by the present invention has good fermentation effect. Clinical trials have shown that the use of animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid can improve the symptoms of anorexia after tumor and related treatments, increase the weight of patients, and improve the quality of life of patients without adverse reactions. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0024] Figure 1 This is a picture of the colony morphology of Bifidobacterium animalis subspecies lactis ZKBb1223.1.

[0025] Figure 2 This is a Gram staining picture of Bifidobacterium animalis subsp. lactis ZKBb1223.1. DETAILED DESCRIPTION

[0026] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0027] Unless otherwise specified, the reagents, materials, and instruments involved in the following examples are all conventional commercially available products; the methods used are all conventional methods in the art unless otherwise specified.

[0028] Among them, mupirocin lithium salt and TOS culture medium were purchased from Qingdao Haibo Biotechnology Co., Ltd., L-cysteine ​​hydrochloride was purchased from Sigma-Aldrich, USA, MRS liquid culture medium and MRS solid culture medium were purchased from Beijing Aoboxing Biotechnology Co., Ltd., DNA extraction kit was purchased from Beijing Tiangen Biochemical Technology Co., Ltd., and ginseng was purchased from Henan Yanhuang Pharmacy.

[0029] The Lactobacillus plantarum RZKLp100 involved in the following examples has been disclosed in Chinese invention patent CN 118374549 A, and Bifidobacterium animalis subsp. lactis BB-12 was purchased from Chr. Hansen of Denmark.

[0030] The ginseng extract involved in the following examples refers to the liquid component obtained by soaking ginseng in water and then boiling it. The specific preparation method is as follows:

[0031] Accurately weigh a certain amount of ginseng and soak it in 8 times the amount of water for 1 hour;

[0032] Put the soaked ginseng for 1 hour and the soaking water into the Chinese medicine steaming pot, boil it with high heat first, and then simmer it with low heat for 90 minutes. Note: the soaking water should be used directly for decocting the medicine without changing the water to avoid wasting the effective ingredients; stir moderately during the decocting process to prevent the pot from sticking, but avoid opening the lid frequently to prevent the loss of volatile ingredients;

[0033] The filtrate was separated by filtration, and then steamed and concentrated until the crude drug content (the amount of ginseng used / the volume of the ginseng extract after concentration) was 1.0 g / mL, thereby obtaining the ginseng extract.

[0034] Example 1 Isolation and identification of Bifidobacterium animalis subspecies lactis ZKBb1223.1

[0035] 1. Sample source: Breast milk collected from lactating women in Jilin City, Jilin Province in December 2009.

[0036] 2. Strain isolation: dilute the collected samples with physiological saline to 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 100 µL of the dilution was taken out and evenly spread on TOS medium containing 0.05 g / L mupirocin lithium salt, and cultured anaerobically in a biochemical incubator at 37 °C for 48 h to observe the growth of the colonies.

[0037] After the colony is formed, pick a single colony and perform three-zone streaking three times, and repeat the streaking separation several times until the colony is purified. The purified strain is inoculated into MRS liquid medium supplemented with 0.05g / L L-cysteine ​​hydrochloride for cultivation, and the culture medium of different strains and 50% sterile glycerol are mixed at a ratio of 1:1 (v / v), named separately, and stored in a -80℃ refrigerator.

[0038] 3. Identification of strains

[0039] (1) Colony morphology identification

[0040] ① Inoculate one of the isolated strains, 1223.1, onto TOS medium containing 0.05 g / L mupirocin lithium salt, culture anaerobically for 48 h, and observe and record the morphology and color of the colonies. Figure 1 As shown, the colonies were milky white, round, moist, opaque and had neat edges.

[0041] ② Pick the colonies on the medium in ① for smearing, fixation, primary staining with crystal violet, mordanting, decolorization, washing with water, secondary staining with safranin, drying, and microscopic examination. The microscopic examination results of strain 1223.1 are shown in Figure 2 Microscopic observation revealed that the bacteria were irregular rod-shaped, slightly swollen at the top, arranged singly or in pairs, and were Gram-positive.

[0042] (2) 16S rDNA sequence determination

[0043] ① Extraction of strain genomic DNA: Take the culture medium of isolated strain 1223.1, centrifuge at 5000r / min for 10min, discard the supernatant after centrifugation, collect the pure culture bacteria to be tested, use a DNA extraction kit to extract the strain genomic DNA, and operate according to the instructions.

[0044] ②PCR amplification: using bacterial universal primers 27F / 1492R.

[0045] The PCR amplification conditions were as follows: pre-denaturation at 94°C for 4 min; denaturation at 94°C for 30 s; annealing at 55°C for 35 s; extension at 72°C for 90 s; after 32 cycles, the final extension was at 72°C for 5 min.

[0046] ③Gene sequencing: PCR products were identified by 1.0% agarose gel electrophoresis, and gene sequencing was performed on the amplified products. The sequencing work was completed by Beijing Qingke Biotechnology Co., Ltd.

[0047] The 16S rDNA sequence of strain 1223.1 is as follows:

[0048]

[0049] After BLAST comparison, it was determined that strain ZKBb1223.1 was Bifidobacterium animalis subsp. lactis, so strain ZKBb1223.1 was named Bifidobacterium animalis subsp. lactis ZKBb1223.1.

[0050] (3) Strain preservation: The strain ZKBb1223.1 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on May 13, 2024. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The deposit number is CGMCC No. 30550 and the classification name is Bifidobacterium animalis subsp. lactis Bifidobacterium animalis subsp. lactis .

[0051] Example 2 Preparation of Ginseng Fermentation Broth of Bifidobacterium animalis subsp. lactis ZKBb1223.1

[0052] 1. Animal Bifidobacterium lactis subsp. ZKBb1223.1 Ginseng fermented liquid

[0053] (1) Pretreatment of ginseng extract:

[0054] Add 4g / 100mL yeast extract and 5g / 100mL fructooligosaccharide to the ginseng extract, mix well, sterilize at 121℃ for 15min, and set aside;

[0055] (2) Preparation of Bifidobacterium animalis subspecies lactis ZKBb1223.1 bacterial suspension:

[0056] The preserved animal Bifidobacterium lactis subsp. ZKBb1223.1 was inoculated into MRS liquid medium supplemented with 0.05 g / L L-cysteine ​​hydrochloride, cultured anaerobically at 37°C for 24 h, activated and propagated to the second generation, and prepared into a bacterial suspension;

[0057] (3) Fermented ginseng extract:

[0058] The bacterial suspension was inoculated into the sterilized ginseng extract at an inoculum rate of 2wt%, and after shaking, the mixture was placed in an anaerobic incubator at 37°C and cultured for 24 hours to obtain animal Bifidobacterium lactis subsp. ZKBb1223.1 ginseng fermentation liquid I.

[0059] 2. Animal Bifidobacterium lactis subsp. ZKBb1223.1 Ginseng fermented liquid II

[0060] (1) Pretreatment of ginseng extract:

[0061] Add 3g / 100mL yeast extract powder and 6g / 100mL fructooligosaccharide to the ginseng extract, mix well, sterilize at 121℃ for 15min, and set aside;

[0062] (2) Preparation of Bifidobacterium animalis subspecies lactis ZKBb1223.1 bacterial suspension:

[0063] The preserved animal Bifidobacterium lactis subsp. ZKBb1223.1 was inoculated into MRS liquid medium supplemented with 0.05 g / L L-cysteine ​​hydrochloride, cultured anaerobically at 37°C for 24 h, activated and propagated to the third generation, and prepared into a bacterial suspension;

[0064] (3) Fermented ginseng extract:

[0065] The bacterial suspension was inoculated into the sterilized ginseng extract at an inoculum rate of 3wt%, and after being shaken, the mixture was placed in an anaerobic incubator at 37°C and cultured for 24 hours to obtain the animal Bifidobacterium lactis subsp. ZKBb1223.1 ginseng fermentation liquid II.

[0066] 3. Animal Bifidobacterium lactis subspecies ZKBb1223.1 Ginseng fermented liquid

[0067] (1) Pretreatment of ginseng extract:

[0068] Add 2g / 100mL yeast extract powder and 7g / 100mL fructooligosaccharide to the ginseng extract, mix well, sterilize at 121℃ for 15min, and set aside;

[0069] (2) Preparation of Bifidobacterium animalis subspecies lactis ZKBb1223.1 bacterial suspension:

[0070] The preserved animal Bifidobacterium lactis subsp. ZKBb1223.1 was inoculated into MRS liquid medium supplemented with 0.05 g / L L-cysteine ​​hydrochloride, cultured anaerobically at 37°C for 24 h, activated and propagated to the third generation, and prepared into a bacterial suspension;

[0071] (3) Fermented ginseng extract:

[0072] The bacterial suspension was inoculated into the sterilized ginseng extract at an inoculum rate of 4wt%, and after being shaken, the mixture was placed in an anaerobic incubator at 37°C and cultured for 24 hours to obtain animal Bifidobacterium lactis subsp. ZKBb1223.1 ginseng fermentation liquid III.

[0073] Comparative Example 1 Preparation of Ginseng Fermentation Broth IV

[0074] (1) Pretreatment of ginseng extract:

[0075] Add 3g / 100mL yeast extract powder and 6g / 100mL fructooligosaccharide to the ginseng extract, mix well, sterilize at 121℃ for 15min, and set aside;

[0076] (2) Preparation of Bifidobacterium animalis subspecies lactis BB-12 bacterial suspension:

[0077] The preserved animal Bifidobacterium lactis subsp. BB-12 was inoculated into MRS liquid medium supplemented with 0.05 g / L L-cysteine ​​hydrochloride, cultured anaerobically at 37° C. for 24 h, activated and propagated to the third generation, and a bacterial suspension was prepared;

[0078] (3) Fermented ginseng extract:

[0079] The bacterial suspension was inoculated into the sterilized ginseng extract at an inoculum rate of 3wt%, and after being shaken well, the mixture was placed in an anaerobic incubator at 37°C and cultured for 24 hours to obtain ginseng fermentation liquid IV.

[0080] Comparative Example 2 Preparation of Ginseng Fermentation Liquid V

[0081] (1) Pretreatment of ginseng extract:

[0082] Add 3g / 100mL yeast extract powder and 6g / 100mL fructooligosaccharide to the ginseng extract, mix well, sterilize at 121℃ for 15min, and set aside;

[0083] (2) Preparation of Lactobacillus plantarum RZKLp100 bacterial suspension:

[0084] The preserved Lactobacillus plantarum RZKLp100 was inoculated into MRS liquid medium, cultured aerobically at 37°C for 24 hours, activated and propagated to the third generation, and a bacterial suspension was prepared;

[0085] (3) Fermented ginseng extract:

[0086] The bacterial suspension was inoculated into the sterilized ginseng extract at an inoculum rate of 3wt%, and after being shaken well, the mixture was placed in an aerobic incubator at 37°C and cultured for 24 hours to obtain ginseng fermentation liquid V.

[0087] Test Case

[0088] The pH values ​​and viable counts of the animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation broth 1, animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation broth 2, animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation broth 3 prepared in Example 2, ginseng fermentation broth 4 prepared in Comparative Example 1, and ginseng fermentation broth 5 prepared in Comparative Example 2 were measured, and the results are shown in Table 1 below.

[0089] Table 1 pH value and viable bacterial count of ginseng fermentation liquid

[0090]

[0091] By comparison, it can be seen that the fermentation effect of the animal Bifidobacterium lactis subspecies ZKBb1223.1 provided by the present invention is better than the existing commonly used animal Bifidobacterium lactis subspecies BB-12 and other strains screened in the laboratory.

[0092] Comparative Example 3 Preparation of Ginseng Composite Bacterial Liquid

[0093] (1) Pretreatment of ginseng extract:

[0094] Add 3g / 100mL yeast extract powder and 6g / 100mL fructooligosaccharide to the ginseng extract, mix well, sterilize at 121℃ for 15min, and set aside;

[0095] (2) Preparation of Bifidobacterium animalis subspecies lactis ZKBb1223.1 bacterial suspension:

[0096] The preserved animal Bifidobacterium lactis subsp. ZKBb1223.1 was inoculated into an MRS liquid culture medium supplemented with 0.05 g / L of L-cysteine ​​hydrochloride, anaerobically cultured at 37° C. for 24 h, activated and propagated to the third generation, and then inoculated into another MRS liquid culture medium supplemented with 0.05 g / L of L-cysteine ​​hydrochloride according to an inoculum amount of 3 wt %, and placed in an anaerobic incubator at 37° C. for continued culture to obtain a bacterial suspension;

[0097] (3) Mixing:

[0098] The bacterial suspension was added to the sterilized ginseng extract to make the concentration of animal Bifidobacterium lactis subsp. ZKBb1223.1 in the liquid reach 2.36×10 9 CFU / mL, and the ginseng composite bacterial liquid was prepared.

[0099] Example 3 Clinical Trial

[0100] 1. Case selection criteria:

[0101] (1) Case inclusion criteria:

[0102] ① Aged 18-80 years old, diagnosed with malignant tumor by pathology or cytology according to the diagnostic criteria of "Chinese Common Malignant Tumor Diagnosis and Treatment Guidelines", with no restrictions on clinical stage and pathological type;

[0103] ② Patients undergoing surgery, radiotherapy or chemotherapy and who have symptoms of anorexia;

[0104] ③Karnofsky functional status score (KPS score)>60 points, no severe heart, liver, kidney, or bone marrow dysfunction;

[0105] ④ The expected survival period is greater than 2 months;

[0106] ⑤ Voluntarily accept the treatment of this program, have good compliance, and sign the informed consent form;

[0107] ⑥ No cognitive or mental disorders, normal language expression ability, and able to cooperate with the examination.

[0108] (2) Case exclusion criteria:

[0109] ① Those who do not meet the inclusion criteria;

[0110] ② Those with cognitive and mental disorders;

[0111] ③ Those who are unable to eat through the mouth or have eating disorders such as dysphagia and intestinal obstruction;

[0112] ④ Pregnant or lactating women;

[0113] ⑤ Those who are participating in other drug trials or taking other appetite-enhancing drugs at the same time, or using oral and / or intravenous nutritional support therapy.

[0114] (3) Case exclusion and dropout criteria

[0115] ① The subject voluntarily requests to withdraw from the trial;

[0116] ② The subjects have poor compliance and cannot complete all the treatment;

[0117] ③ Those who experience severe adverse reactions during treatment;

[0118] ④ Patients whose condition worsened or died during the study period.

[0119] 2. Experimental plan:

[0120] (1) Trial grouping: All 160 patients were randomly assigned to the ZKBb1223.1 fermented ginseng treatment group, BB-12 fermented ginseng treatment group, RZKLp100 fermented ginseng treatment group, and ginseng compound bacterial liquid treatment group, with 40 cases in each group. There was no significant difference in age, gender, appetite, KPS score, body weight, bone marrow function, liver and kidney function, and electrocardiogram changes among the groups (P>0.05), and the groups were comparable.

[0121] (2) Treatment method: Each person in the ZKBb1223.1 fermented ginseng treatment group took 50 mL of the animal Bifidobacterium lactis subsp. ZKBb1223.1 ginseng fermentation liquid II of Example 2 three times a day, once in the morning, noon and evening, for a course of 14 days;

[0122] In the BB-12 fermented ginseng treatment group, each person took the ginseng fermented liquid IV prepared in comparative example 1 three times a day, 50 mL each time, once in the morning, noon and evening, and the treatment course lasted for 14 days;

[0123] Each person in the RZKLp100 fermented ginseng treatment group took ginseng fermented liquid V prepared in comparative example 2 three times a day, 50 mL each time, once in the morning, noon and evening, and the treatment course lasted for 14 days;

[0124] Each person in the ginseng compound bacteria liquid treatment group took the ginseng compound bacteria liquid prepared in Comparative Example 3 three times a day, 50 mL each time, once in the morning, noon and evening, and took it immediately after preparation. The treatment course lasted for 14 days.

[0125] 3. Observation indicators and results:

[0126] (1) Body weight: Observe and record the body weight before and 14 days after treatment. Refer to the weight change in the 2002 edition of the "Guidelines for Clinical Research of New Chinese Medicines" as the evaluation standard.

[0127] Markedly effective: body weight after treatment increased by ≥1kg compared with that before treatment;

[0128] Stable: weight gain or loss after treatment is less than 1 kg;

[0129] Ineffective: Body weight reduction after treatment is ≥1kg compared with that before treatment.

[0130] The results are shown in Table 2 below.

[0131] Table 2 Comparison of the efficacy evaluation of weight changes in each group before and after treatment (unit: case)

[0132]

[0133] (2) Appetite: According to the fifth edition of the Common Criteria for Adverse Events (CTCAE), appetite loss is divided into 5 levels:

[0134] Grade I: Loss of appetite, but unchanged eating habits;

[0135] Grade II: Changes in eating habits, not accompanied by weight loss or malnutrition, but requiring oral nutritional supplements;

[0136] Grade III: Significant weight loss or malnutrition symptoms (such as insufficient oral intake of calories and / or fluids), requiring nasogastric feeding or total parenteral nutrition;

[0137] Grade IV: life-threatening, requiring emergency treatment;

[0138] Level V: Death.

[0139] Efficacy evaluation: after treatment, the symptoms dropped by 2 or more grades compared with before treatment, or dropped to grade 0 (no loss of appetite) for marked effect; after treatment, the symptoms dropped by 1 grade compared with before treatment for effective; the symptoms did not alleviate or worsened for ineffective. Total effective rate (%) = (marked effect + effective) / total number of cases × 100%.

[0140] The results are shown in Table 3 below.

[0141] Table 3 Comparison of the evaluation of appetite efficacy in each group before and after treatment (Unit: case)

[0142]

[0143] (3) Quality of life: assessed using the Karnofsky Performance Score (KPS).

[0144] The KPS score decreased by ≥10 points compared with before treatment;

[0145] The KPS score decreased or increased by <10 points compared with before treatment and was stable;

[0146] The KPS score was improved when it increased by ≥10 points compared with before treatment.

[0147] Among them, the total effective rate (%) = (number of stable cases + number of improved cases) / total number of cases × 100%.

[0148] The results are shown in Table 4 below.

[0149] Table 4 Comparison of KPS scores in each group before and after treatment (unit: case)

[0150]

[0151] (4) Safety observation: Blood routine, urine routine, stool routine, liver and kidney function tests were performed before treatment and after the treatment.

[0152] By comparison, all patients in this trial had no adverse reactions during the treatment.

[0153] 4. Conclusion:

[0154] First, from the evaluation of the efficacy of weight change, the number of cases of marked effect in the ZKBb1223.1 fermented ginseng treatment group was significantly higher than that in the BB-12 fermented ginseng treatment group, the RZKLp100 fermented ginseng treatment group, and the ginseng compound bacterial liquid treatment group, with a marked effect rate of 62.5%, significantly better than the other groups of 45.0%, 42.5%, and 40.0%. This data strongly proves the significant effect of ZKBb1223.1 fermented ginseng extract in promoting patients' weight recovery.

[0155] Secondly, in terms of appetite efficacy evaluation, the ZKBb1223.1 fermented ginseng treatment group also performed well, with the sum of significant and effective cases reaching 37 cases, accounting for 92.5% of the total cases, much higher than 70.0%, 62.5% and 52.5% of other groups, confirming that the fermented ginseng liquid of the present invention significantly improved the patient's appetite.

[0156] Finally, in the comparison of KPS scores, the ZKBb1223.1 fermented ginseng treatment group also showed obvious advantages, with 26 cases of improvement, accounting for 90.0% of the total number of cases, significantly better than the 72.5%, 70.0% and 65.0% of the other groups. KPS score is one of the important indicators to measure the quality of life of cancer patients. The significant advantage of the ZKBb1223.1 fermented ginseng treatment group in this indicator further illustrates the positive role of the fermented ginseng liquid of the present invention in improving the quality of life of patients.

[0157] Analysis of the differences in data among the ZKBb1223.1 fermented ginseng treatment group, BB-12 fermented ginseng treatment group, and RZKLp100 fermented ginseng treatment group showed that the animal Bifidobacterium lactis subspecies ZKBb1223.1 provided by the present invention has a more efficient fermentation ability for ginseng extract, which not only ensures a significant increase in the number of live bacteria in the fermentation product, but also may promote the deep release and efficient utilization of the active ingredients in ginseng.

[0158] At the same time, the difference between the data of the ZKBb1223.1 fermented ginseng treatment group and the ginseng composite bacterial liquid treatment group was analyzed, and it was found that animal Bifidobacterium lactis subspecies ZKBb1223.1 was likely to produce metabolites and other components that help improve the appetite of patients undergoing tumor radiotherapy and chemotherapy during the fermentation process, thereby further enhancing the therapeutic effect. It can be seen that the present invention uses animal Bifidobacterium lactis subspecies ZKBb1223.1 to ferment ginseng extract, and successfully achieves the synergistic effect of two natural ingredients, microorganisms and ginseng, compared with simple mixing, and has achieved the significant effect of "1+1>2" in practical applications.

[0159] Example 4 Typical Case

[0160] Case 1: Guan, female, housewife, went to the doctor for irregular vaginal bleeding accompanied by pain in the lower abdomen or lumbosacral region and was diagnosed with cervical cancer. After 4 chemotherapy sessions, she had a loss of appetite, could hardly eat any food, and had a serious weight loss. After drinking the animal bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid II of this embodiment 2, her appetite was significantly improved, and she could eat some porridge, soup and the like; she continued to drink the animal bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid II in embodiment 2 of the present invention, and her appetite gradually recovered, her spirit gradually recovered, her resistance also increased, and her quality of life improved.

[0161] Case 2: Liu, male, employee, 31 years old, initially had symptoms such as blood in sputum after nasal suction, or bloody nose, but did not pay attention to it. Later, lumps appeared on both sides of the neck, which were hard and had poor mobility. A physical examination in the hospital diagnosed nasopharyngeal carcinoma. After chemotherapy, he suffered from fatigue, dizziness, loss of appetite, nausea, vomiting, tasteless or changed taste in the mouth, insomnia or drowsiness, etc. After taking animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid II of Example 2 of the present invention three times in the morning, noon and evening, it was found that the symptoms of nausea and vomiting were reduced; appetite increased, dizziness and fatigue symptoms were reduced, and appetite improved significantly after drinking for one month, weight increased, and sleep quality also improved.

[0162] Case 3: Li, male, 50 years old, working in the army, has liked smoking since childhood, even 5 packs of cigarettes a day, never leaving his cigarettes, unable to quit. After chemotherapy, he had anorexia, mental fatigue, and weight loss. After drinking the animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid II of Example 2 of the present invention, his appetite was significantly improved, he could eat some food such as porridge and soup, and he could get out of bed and walk; continuing to drink the animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid II in Example 2 of the present invention, his appetite gradually recovered, his spirit gradually recovered, and his weight also recovered.

[0163] Case 4: Liu, male, self-employed, had hepatitis B, and had poor diet and living habits, which led to liver cancer. After one cycle of chemotherapy, his hair almost fell out, and he developed symptoms such as fatigue, weight loss, loss of appetite, abdominal distension, diarrhea, nausea, and vomiting. He had poor resistance and was prone to illness. After taking the animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermented liquid II of Example 2 of the present invention three times a week, morning, noon, and evening, it was found that the symptoms of nausea and vomiting subsided and the appetite increased. After drinking for one month, his appetite improved significantly, his weight increased, and his immunity also improved accordingly.

[0164] Case 5: Chen, female, teacher, went to the doctor for blood in stool, upper abdominal pain, and difficulty in defecation and was diagnosed with colon cancer. After admission for examination, she underwent laparoscopic colectomy and postoperative adjuvant chemotherapy. After chemotherapy, she had anorexia, listlessness, sallow complexion, fatigue, weak voice, and weight loss. After taking the animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid II of Example 2 of the present invention three times in the morning, noon, and evening for one week, it was found that her appetite increased. After drinking for one month, her appetite improved significantly, her weight increased, her mental outlook improved, and her complexion became ruddy.

[0165] Although the present invention has been described in detail with reference to the accompanying drawings and in combination with the preferred embodiments, the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and these modifications or substitutions shall be within the scope of the present invention. Any person of ordinary skill in the art may easily think of changes or substitutions within the technical scope disclosed by the present invention, and these shall be within the scope of protection of the present invention.

Claims

1. A ginseng fermented liquid of Bifidobacterium lactis subspecies ZKBb1223.1 capable of improving the appetite of patients undergoing radiotherapy and chemotherapy for tumors, characterized in that: Prepared by the following method: (1) Soaking ginseng in water and then boiling it to obtain ginseng extract; (2) Add yeast extract powder and oligofructose to the ginseng extract and sterilize for later use; (3) inoculating the ginseng extract sterilized in step (2) with Bifidobacterium animalis subsp. lactis ( Bifidobacterium animalis subsp. lactis )ZKBb1223.1, shake well and culture anaerobically at 37℃ to obtain; Bifidobacterium animalis subsp. lactis ZKBb1223.1 was deposited at the General Microbiology Center of China Microbiological Culture Collection Administration on May 13, 2024. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The deposit number is CGMCC No. 30550. The classification name is Bifidobacterium animalis subsp. lactis Bifidobacterium animalis subsp. lactis .

2. The animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid as claimed in claim 1, characterized in that In step (1), the ginseng is soaked in water of eight times its mass for 60 minutes.

3. The animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid as claimed in claim 1, characterized in that In step (1), the boiling condition is to boil the mixture and then keep boiling for 90 minutes.

4. The animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid as claimed in claim 1, characterized in that In step (1), after decoction, the filtrate is filtered and separated, and the ginseng extract with a crude drug content of 1.0 g / mL is obtained by decoction and concentration.

5. The animal Bifidobacterium lactis subspecies ZKBb1223.1 ginseng fermentation liquid according to claim 1, characterized in that: In step (2), based on the volume of the ginseng extract, the amount of yeast extract powder added is 2-4 g / 100 mL, and the amount of oligofructose added is 5-7 g / 100 mL.

6. The ginseng fermentation liquid of animal Bifidobacterium lactis subspecies ZKBb1223.1 according to claim 1, characterized in that: In step (3), 2 wt% to 4 wt% of a bacterial suspension of Bifidobacterium animalis subspecies lactis ZKBb1223.1 is inoculated into the ginseng extract, and the bacterial suspension is prepared by the following method: Animal Bifidobacterium lactis subsp. ZKBb1223.1 was inoculated into MRS liquid culture medium supplemented with 0.05 g / L L-cysteine ​​hydrochloride, cultured anaerobically at 37°C for 24 h, activated and propagated to the 2nd to 3rd generation, and prepared into a bacterial suspension.

7. The ginseng fermentation liquid of animal Bifidobacterium lactis subspecies ZKBb1223.1 according to claim 1, characterized in that: The pH of ginseng fermentation liquid of Bifidobacterium animalis subspecies Lactis ZKBb1223.1 is 3.5~3.

9.

8. Use of the ginseng fermented liquid of Bifidobacterium animalis subsp. lactis ZKBb1223.1 as claimed in any one of claims 1 to 7 in the preparation of a medicament for improving the appetite of patients undergoing radiotherapy and chemotherapy for tumors.

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