A compound preparation for nourishing and maintaining the vagina and pelvic floor muscles by using plant fermented lactic acid bacteria lysate

Abstract

This invention discloses a compound preparation of plant-fermented lactic acid bacteria lysate for nourishing and protecting the vaginal pelvic floor muscles. By weight, it comprises the following components: kidney bean-honeysuckle fruit compound ferment: 4-8 parts; pomegranate peel-derived lactic acid bacteria lysate: 5-9 parts; bamboo fungus mycelium extract: 3-7 parts; compound sophorolipid: 6-8 parts; sodium hyaluronate: 2-5 parts; total glycosides of Centella asiatica: 1-3 parts; glycerin: 5-10 parts; lactic acid: 1-2 parts; purified... Water: 36-45 parts; Carbomer 940: 2-4 parts; Trehalose: 1-3 parts; Chitosan: 1-3 parts; Methylchloroisothiazolinone: 0.1-0.3 parts; Sodium hydroxide: 0.1-0.2 parts; This invention can regulate the vaginal microecological balance, nourish the vaginal mucosa, enhance the toughness of pelvic floor muscle fibers, and promote the repair of pelvic floor muscle damage, solving the technical problems of existing preparations having single function, weak targeted pelvic floor muscle care, and easy disruption of the vaginal microecology.

Description

Technical Field

[0001] This invention belongs to the field of female reproductive health care technology, specifically a compound preparation of plant fermented lactic acid bacteria cell lysate to nourish and care for the vaginal pelvic floor muscles. Background Technology

[0002] A woman's vaginal and pelvic floor muscle health is a crucial component of reproductive health. The pelvic floor muscles act like a "net," supporting organs such as the urethra, bladder, uterus, and rectum. Hormonal changes during pregnancy, the weight of the fetus, and muscle stretching during childbirth can easily lead to pelvic floor muscle relaxation, resulting in problems such as urinary incontinence, vaginal laxity, and uterine prolapse. Simultaneously, an imbalance in the vaginal microecology can easily cause vaginitis and mucosal damage, further exacerbating pelvic floor muscle dysfunction. Currently, most feminine hygiene products on the market focus on vaginal cleansing or simply pelvic floor muscle repair, offering limited functionality and failing to achieve the synergistic effect of vaginal microecological regulation and pelvic floor muscle nourishment and repair.

[0003] In existing technologies, lactic acid bacteria lysate products are widely used in skin care and intestinal health. They are rich in probiotic metabolites, amino acids, peptides and other active ingredients, and have good nourishing and repairing effects. However, when applied to vaginal and pelvic floor muscle care, there are problems such as low utilization rate of active ingredients and weak targeting. At the same time, existing pelvic floor muscle care preparations mostly use chemical ingredients or single plant extracts, which can easily irritate the vaginal mucosa, disrupt the vaginal microecological balance, and are difficult to effectively enhance the toughness of pelvic floor muscle fibers and promote damage repair.

[0004] Furthermore, existing plant fermentation products mostly focus on fruit and vegetable blends or single plant fermentation, with few reports on the use of fermented legumes and small berries for vaginal pelvic floor muscle care. While ingredients like sophorolipids and bamboo extracts have antibacterial and repairing effects, they are primarily used in washes or cosmetics; there are no reports of their synergistic use with lactic acid bacteria lysates and complex plant ferments for vaginal pelvic floor muscle care. Therefore, developing a compound preparation that combines vaginal microecological regulation, vaginal mucosal nourishment, and pelvic floor muscle repair functions, while also being highly safe and non-irritating, has become an urgent need in the field of women's feminine hygiene.

[0005] Based on this, a compound preparation of plant-fermented lactic acid bacteria lysate for nourishing and maintaining vaginal pelvic floor muscles was designed. Summary of the Invention

[0006] In view of the above situation and to overcome the defects of the prior art, the present invention provides a compound preparation of plant fermented lactic acid bacteria lysate to nourish and maintain the vaginal pelvic floor muscles, which effectively solves the problems mentioned in the background.

[0007] To achieve the above objectives, the present invention provides the following technical solution: a compound preparation for nourishing and protecting the vaginal pelvic floor muscles using plant-fermented lactic acid bacteria lysate, comprising the following components by weight: Kidney bean-honeysuckle berry fermented compound: 4-8 parts; Pomegranate peel-derived lactic acid bacteria lysate: 5-9 parts; Bamboo fungus mycelium extract: 3-7 parts; Complex sophorolipids: 6-8 parts; Sodium hyaluronate: 2-5 parts; Total glycosides from Centella asiatica: 1-3 parts; Glycerin: 5-10 parts; Lactic acid: 1-2 parts; Purified water: 36-45 parts; Carbomer 940: 2-4 servings; Trehalose: 1-3 parts; Chitosan: 1-3 parts; Methylchloroisothiazolinone: 0.1-0.3 parts; Sodium hydroxide: 0.1-0.2 parts.

[0008] Preferably, the preparation method of the kidney bean-honeysuckle fruit complex ferment is as follows: S1.1 Raw material pretreatment: Take 200-300 grams of fresh kidney beans and 150-250 grams of fresh honeysuckle berries. Remove impurities from the kidney beans, wash them, and cut them into 1×1 cm pieces. Remove the stems from the honeysuckle berries, wash them, mix the two together, put them into a juicer, add 500-700 ml of purified water, and juice for 10-15 minutes to obtain kidney bean-honeysuckle berry mixed juice. S1.2 Sterilization treatment: Pour the mixed juice into the fermentation tank, adjust the initial pH value to 4.8-5.2, sterilize at 115-120℃ for 10-15 minutes, cool to 32-36℃ to obtain the fermentation substrate; S1.3 Inoculation and Fermentation: Inoculate the fermentation substrate with a compound lactic acid bacteria at a volume of 2%-3% of the substrate. The compound lactic acid bacteria is a mixture of *Lactobacillus plantarum* and *Lactobacillus acidophilus*, with a volume ratio of 1:1. Ferment at 34-36℃ under anaerobic conditions for 24-30 hours, stirring every 6 hours at a speed of 120-150 rpm. The *Lactobacillus plantarum* is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1295 and deposit date of January 10, 2025; the *Lactobacillus acidophilus* is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1854 and deposit date of January 10, 2025. S1.4 Post-processing: After fermentation, centrifuge the fermentation broth at 5000-6000 rpm for 10-15 minutes, take the supernatant, filter it through a 0.22-micron filter membrane to obtain the kidney bean-blue honeysuckle berry compound fermentation product, and store it at 4℃ for later use. Preferably, the method for preparing the pomegranate peel-derived lactic acid bacteria lysate is as follows: S2.1 Preparation of pomegranate peel culture medium: Take 100-150 grams of dried pomegranate peel, crush it and pass it through an 80-100 mesh sieve, add 500-700 ml of purified water, adjust the pH value to 3.0-4.0, the pomegranate peel mass concentration is 3-7 g / ml, sterilize at 115-120℃ for 10-15 minutes, cool to 30-37℃, and obtain pomegranate peel fermentation culture medium; S2.2, Lactic acid bacteria proliferation: Inoculate Lactobacillus plantarum (preservation information is the same as in step S1.3) into the pomegranate peel fermentation medium. The inoculation amount is 4%-8% of the medium volume. Incubate at 30-37℃ for 48-72 hours to achieve lactic acid bacteria proliferation. S2.3. Cell isolation: After the proliferation is completed, first filter to remove pomegranate peel residue, then centrifuge the remaining liquid at a speed of 5000-6000 rpm for 5-15 minutes, take the precipitate, and obtain Lactobacillus plantarum cells; S2.4, Cell lysis treatment: Add Lactobacillus plantarum cells to 200-300 ml of physiological saline, stir well, place in an ice bath environment, and use ultrasonic disruption method to perform cell lysis. The ultrasonic power is 250-300 watts, with a 1-second interval between every 2 seconds of operation, and the disruption time is 25-30 minutes. S2.5 Purification: After lysing, centrifuge at 5000-6000 rpm for 10-15 minutes, take the supernatant, filter through a 0.22-micron filter membrane, freeze dry under vacuum at -40 to -30℃ for 12-18 hours to obtain pomegranate peel-derived lactic acid bacteria lysate powder. Preferably, the preparation method of the bamboo yellow mycelium extract is as follows: S3.1 Mycelial Culture: Take bamboo yellow fungus, inoculate it into PDA medium, and culture it for 7-10 days at 25-28℃ and 60%-70% humidity to obtain bamboo yellow mycelium; PDA medium formula: 200g potato, 20g glucose, 15-20g agar, 1000ml purified water, sterilize at 121℃ for 20 minutes; S3.2 Pretreatment: Take out the bamboo yellow mycelium, rinse it with purified water 3-5 times, dry it at 35℃ to constant weight, pulverize it and pass it through an 80-mesh sieve to obtain bamboo yellow mycelium powder; S3.3 Extraction: Take 30-50 grams of bamboo yellow mycelium powder, add 400-600 ml of anhydrous ethanol, with a material-to-liquid ratio of 1:13 to 1:15, and extract at 30-35℃ for 20-24 hours, stirring once every 4 hours during the process, with a stirring speed of 100-120 rpm. S3.4 Purification: After extraction, centrifuge at 4000-5000 rpm for 10-12 minutes, collect the supernatant, and distill the supernatant under reduced pressure at 50-60℃ and 0.06-0.08 MPa to remove ethanol, obtain a concentrated solution, filter through a 0.22-micron filter membrane, and freeze-dry under vacuum to obtain bamboo yellow mycelium extract; Preferably, the preparation method of the complex sophorolipid is as follows: S4.1, Strain Activation: Candida albicans was inoculated into an activation medium and cultured with shaking at 28-32℃ and 150-180 rpm for 18-24 hours to obtain the activated strain. The activation medium formula was: 20-30 g glucose, 5-10 g yeast extract, 5-10 g peptone, and 1000 ml purified water, sterilized at 121℃ for 20 minutes. The Candida albicans strain was deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 2.538 and deposit date of January 10, 2025. S4.2 Fermentation culture: Inoculate the activated strain into the fermentation medium at a volume of 5%-8% of the fermentation medium. The fermentation medium consists of 80-120g glucose, 50-80g coconut oil, 3-5g yeast extract, 1-2g potassium dihydrogen phosphate, 0.5-1g magnesium sulfate, and 1000ml purified water. Adjust the pH to 5.5-6.5 and ferment with shaking at 28-32℃ and 150-180 rpm for 48-72 hours. S4.3 Separation and purification: After fermentation, centrifuge the fermentation broth at 5000-6000 rpm for 10-15 minutes, take the supernatant, adjust the pH value to 2.0-2.5 with hydrochloric acid, let it stand for 2-3 hours, precipitate, collect the precipitate, wash it 2-3 times with anhydrous ethanol, and vacuum dry it at 40-50℃ for 8-12 hours to obtain the complex sophorolipid.

[0009] A method for preparing a compound preparation of plant-fermented lactic acid bacteria lysate for nourishing and protecting the vaginal pelvic floor muscles includes the following steps: S1: Weigh each component according to the mass fraction, mix the kidney bean-blue honeysuckle fruit complex ferment, pomegranate peel-derived lactic acid bacteria cell lysate, bamboo yellow mycelium extract, and complex sophorolipid, add 20-30 ml of purified water, stir evenly, and stir at a constant temperature of 30-35℃ for 30-40 minutes to obtain mixture A; S2: Add sodium hyaluronate, total asiaticoside, and glycerin to mixture A, stir at 150-200 rpm for 20-30 minutes until completely dissolved to obtain mixture B; S3: Add carbomer 940, trehalose, and chitosan, and continue stirring for 15-20 minutes. Adjust the stirring speed to 120-150 rpm and stir until the system is uniform and free of particles to obtain mixture C. S4: Add lactic acid and the remaining purified water, stir well, adjust the pH of the system to 3.8-4.5 with sodium hydroxide, add methylchloroisothiazolinone, and continue stirring for 10-15 minutes to obtain mixture D; S5: Sterilize the mixture D at 80-85℃ for 20-30 minutes, cool to room temperature, fill and seal to obtain the compound preparation.

[0010] Preferably, in step S5, the rate of cooling to room temperature after sterilization is 2-3℃ / minute, the filling environment is a sterile environment, and the contents are sealed and stored at a temperature of 2-8℃ after filling.

[0011] Preferably, the compound preparation is used in the preparation of vaginal and pelvic floor muscle care products.

[0012] Compared with the prior art, the beneficial effects of the present invention are: 1. This invention identifies four synergistic components through screening: kidney bean-honeysuckle fruit complex fermentation product, pomegranate peel-derived lactic acid bacteria cell lysate, bamboo fungus mycelium extract, and complex sophorolipid. There is a clear synergistic effect among these four components. Combined with other components such as sodium hyaluronate, centella asiatica total glycosides, and glycerin, a triple care system of "microecological regulation - mucosal nourishment - pelvic floor muscle repair" is formed. This solves the technical problems of existing preparations having single function, weak targeting of pelvic floor muscle care, and easy disruption of vaginal microecology, thus achieving synergistic care of the vagina and pelvic floor muscles. 2. The kidney bean-blue honeysuckle fruit compound fermentation product, pomegranate peel-derived lactic acid bacteria cell lysate, bamboo yellow mycelium extract, and compound sophorolipid of the present invention are prepared using natural plant raw materials and probiotic fermentation. The preparation process is green and environmentally friendly, without harmful additives, and the products have high activity and can be mass-produced. 3. The compound preparation of the present invention has high safety and is non-irritating. The pH value is controlled at 3.8-4.5, which is consistent with the acidic environment of the vagina. It can promote the colonization of lactobacilli, maintain the vaginal microecological balance, nourish the vaginal mucosa, repair damage, and enhance the toughness of pelvic floor muscle fibers. It is suitable for long-term vaginal and pelvic floor muscle care and has broad application prospects. Detailed Implementation

[0013] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0014] This invention provides a compound preparation for nourishing and protecting the vaginal pelvic floor muscles by plant-fermented lactic acid bacteria lysate, comprising the following components by mass: kidney bean-honeysuckle fruit compound fermentation product: 4-8 parts; Pomegranate peel-derived lactic acid bacteria lysate: 5-9 parts; Bamboo fungus mycelium extract: 3-7 parts; Complex sophorolipids: 6-8 parts; Sodium hyaluronate: 2-5 parts; Total glycosides from Centella asiatica: 1-3 parts; Glycerin: 5-10 parts; Lactic acid: 1-2 parts; Purified water: 36-45 parts; Carbomer 940: 2-4 servings; Trehalose: 1-3 parts; Chitosan: 1-3 parts; Methylchloroisothiazolinone: 0.1-0.3 parts; Sodium hydroxide: 0.1-0.2 parts.

[0015] This invention utilizes four synergistic components: kidney bean-honeysuckle fruit complex fermentation product, pomegranate peel-derived lactic acid bacteria lysate, bamboo fungus mycelium extract, and complex sophorolipids. A clear synergistic mechanism exists among these four components: the complex sophorolipids, as a natural biosurfactant, gently cleanses the vagina and inhibits harmful bacteria. Simultaneously, it disrupts the lipid barrier on the vaginal mucosa, promoting the penetration of polyphenols and flavonoids from the kidney bean-honeysuckle fruit complex fermentation product, as well as active peptides and amino acids from the pomegranate peel-derived lactic acid bacteria lysate, into the deep layers of the vaginal mucosa and pelvic floor muscles; bamboo fungus mycelium… The extract contains bamboo red fungus extract, which promotes collagen production and enhances the resilience of pelvic floor muscle fibers. Its anti-inflammatory effect also alleviates vaginal mucosal inflammation, creating a suitable environment for the regulation of the microecology by lactobacillus lysate. The active ingredients in pomegranate peel-derived lactobacillus lysate promote pelvic floor muscle cell proliferation and regulate vaginal lactobacillus colonization. The kidney bean-honeysuckle fruit complex ferment nourishes the vaginal mucosa and strengthens the mucosal barrier, further enhancing the microecological regulation effect. These four components work synergistically to form a triple-care system of "microecological regulation - mucosal nourishment - pelvic floor muscle repair." These four components, combined with sodium hyaluronate, total asiatica glycosides, glycerin, lactic acid, and other ingredients, maintain the vaginal acidic environment (pH 3.8-4.5), promote lactobacillus colonization, nourish the vaginal mucosa, repair damage, enhance the resilience of pelvic floor muscle fibers, and promote pelvic floor muscle injury repair. This addresses the problem of single-function existing preparations and is highly safe and non-irritating, suitable for long-term care.

[0016] Sodium hyaluronate has excellent moisturizing and repairing abilities, which can promote the regeneration and repair of vaginal mucosa and reduce inflammation and discomfort; total glycosides of Centella asiatica can promote collagen synthesis, enhance the toughness of pelvic floor muscle fibers, and assist in the repair of pelvic floor muscles; glycerin plays a moisturizing role and maintains the moisture of vaginal mucosa; lactic acid is used to adjust the pH value of the preparation to make it conform to the acidic environment of the vagina, promote the colonization of lactobacilli, and maintain the vaginal microecological balance; purified water is used as a solvent to ensure that all components are fully dissolved and free from impurities.

[0017] Carbomer 940 serves as the gel matrix, enabling the formulation to form a uniform and stable gel, facilitating its application and adhesion to the vaginal and pelvic floor muscle surfaces. Trehalose acts as a protectant, safeguarding the stability of active ingredients such as lactic acid bacteria lysates and preventing their inactivation. Chitosan possesses antibacterial, hemostatic, and mucosal repair properties, which can enhance the formulation's protective effects. Methylchloroisothiazolinone acts as a preservative, extending the formulation's shelf life. Sodium hydroxide is used to adjust the pH of the formulation to a suitable range.

[0018] The compound sophorolipid is a mixture of acidic and lactone-type sophorolipids, with acidic sophorolipids accounting for 60%-70% and lactone-type sophorolipids accounting for 30%-40%. This ratio of compound sophorolipids exhibits both good antibacterial activity and biocompatibility, effectively inhibiting harmful bacteria while promoting the penetration of active ingredients, and is non-irritating to the vaginal mucosa. The bamboo mycelium extract contains ≥0.5% bamboo red mycelium A, ensuring its antioxidant, anti-inflammatory, and repairing effects. The pomegranate peel-derived lactic acid bacteria lysate contains ≥15mg / g of lactic acid bacteria active ingredients, ensuring its microecological regulation and pelvic floor muscle repair effects.

[0019] The preparation method of the kidney bean-honeysuckle berry complex ferment is as follows: S1.1 Raw Material Pretreatment: Take 200-300g of fresh kidney beans and 150-250g of fresh honeysuckle berries. Remove impurities from the kidney beans, wash them, and cut them into 1×1cm pieces. Remove the stems from the honeysuckle berries and wash them. Mix the two together and put them into a juicer. Add 500-700ml of purified water and juice for 10-15 minutes to obtain a kidney bean-honeysuckle berry mixed juice. In this step, the ratio of kidney beans to honeysuckle berries is optimized to fully exert their synergistic effect and increase the content of active ingredients in the fermentation products. The juicing time is controlled at 10-15 minutes to ensure that the raw materials are fully broken down and the active ingredients are released, while avoiding excessive impurities caused by over-juicing.

[0020] S1.2 Sterilization: Pour the mixed juice into a fermentation tank, adjust the initial pH to 4.8-5.2, sterilize at 115-120℃ for 10-15 minutes, and cool to 32-36℃ to obtain the fermentation substrate. Controlling the sterilization temperature and time within the above range effectively kills unwanted microorganisms in the mixed juice, preventing contamination and ensuring the fermentation effect is not affected, while also preserving the nutrients in the raw materials. The cooling temperature of 32-36℃ meets the fermentation temperature requirements of the compound lactic acid bacteria, facilitating subsequent inoculation and fermentation.

[0021] S1.3 Inoculation and Fermentation: Inoculate the fermentation substrate with a compound lactic acid bacteria culture at a volume of 2%-3% of the substrate. The compound lactic acid bacteria culture is a mixture of *Lactobacillus plantarum* and *Lactobacillus acidophilus* in a volume ratio of 1:1. Fermentation is carried out at 34-36℃ under anaerobic conditions for 24-30 hours. During fermentation, the bacteria are stirred every 6 hours at a speed of 120-150 rpm. Maintaining an inoculation volume of 2%-3% ensures a sufficient number of lactic acid bacteria, promoting smooth fermentation. The 1:1 ratio of the mixed lactic acid bacteria allows for synergistic fermentation, enhancing the activity of the fermentation product. Maintaining a fermentation temperature of 34-36℃ and anaerobic conditions promotes the proliferation and metabolism of the lactic acid bacteria. Stirring ensures sufficient contact between the lactic acid bacteria and the fermentation substrate, improving fermentation efficiency and product uniformity. The *Lactobacillus plantarum* strain is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1295 and deposit date of January 10, 2025; the *Lactobacillus acidophilus* strain is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1854 and deposit date of January 10, 2025. The strains can be obtained directly from the depository institutions, facilitating reproduction by those skilled in the art.

[0022] S1.4 Post-processing: After fermentation, centrifuge the fermentation broth at 5000-6000 rpm for 10-15 minutes, collect the supernatant, and filter it through a 0.22-micron filter membrane to obtain the kidney bean-honeysuckle berry complex fermented product. Store at 4℃ for later use. Controlling the centrifugation speed and time within the above range effectively removes impurities and bacterial precipitates from the fermentation broth. The 0.22-micron filter membrane further filters out fine impurities, ensuring the purity of the fermented product. Storing at 4℃ maintains the activity of the fermented product and prevents the inactivation of active ingredients.

[0023] The preparation method of pomegranate peel-derived lactic acid bacteria lysate is as follows: S2.1 Preparation of pomegranate peel culture medium: Take 100-150 grams of dried pomegranate peel, crush it, and pass it through an 80-100 mesh sieve. Add 500-700 ml of purified water, adjust the pH to 3.0-4.0, and the pomegranate peel concentration to 3-7 g / ml. Sterilize at 115-120℃ for 10-15 minutes, and cool to 30-37℃ to obtain the pomegranate peel fermentation culture medium. Crushing the pomegranate peel and passing it through an 80-100 mesh sieve increases its contact area with the culture medium, releasing more active ingredients and providing nutrients for the proliferation of lactic acid bacteria. Adjusting the pH to 3.0-4.0 meets the growth requirements of *Lactobacillus plantarum* and inhibits the growth of other bacteria. The sterilization conditions are the same as in step 1.2 to ensure the sterility of the culture medium.

[0024] S2.2 Lactic acid bacteria proliferation: Inoculate *Lactobacillus plantarum* (preservation information same as step S1.3) into the pomegranate peel fermentation medium at an inoculation rate of 4%-8% of the medium volume. Incubate at 30-37℃ for 48-72 hours to achieve lactic acid bacteria proliferation. Controlling the inoculation rate at 4%-8% ensures rapid proliferation of lactic acid bacteria, and controlling the incubation time at 48-72 hours allows the lactic acid bacteria to reach a stable proliferation state and accumulate sufficient active ingredients. This incubation time was determined through optimization experiments; after 48 hours of incubation, the number of lactic acid bacteria reached 10... 9 When the concentration of CFU / mL was above a certain level, there was no significant increase in the number of cells after 72 hours, thus establishing this range.

[0025] S2.3. Cell Isolation: After proliferation, first filter to remove pomegranate peel residue, then centrifuge the remaining liquid at 5000-6000 rpm for 5-15 minutes, collect the precipitate, and obtain *Lactobacillus plantarum* cells. Filtering to remove pomegranate peel residue avoids impurities affecting subsequent cell lysis treatment. Controlling the centrifugation speed and time within the above range can effectively separate the lactic acid bacteria cells and ensure the purity of the cells.

[0026] S2.4, Cell Lysis Treatment: Add *Lactobacillus plantarum* cells to 200-300 ml of physiological saline, stir well, and place in an ice bath environment. Perform cell lysis using ultrasonic disruption. The ultrasonic power is 250-300 watts, with a 1-second pause after every 2 seconds of operation, for a total disruption time of 25-30 minutes. Physiological saline maintains the osmotic pressure of the cells, preventing the inactivation of active ingredients during cell lysis. The ice bath environment lowers the temperature during ultrasonication, preventing denaturation of active ingredients due to high temperatures. Controlling the ultrasonic power, pause time, and disruption time within the above range ensures thorough cell disruption, releasing intracellular active ingredients while avoiding over-disruption that could damage them. Experiments have verified that the cell disruption rate reaches over 90% under these parameters.

[0027] S2.5 Purification: After lysing, centrifuge at 5000-6000 rpm for 10-15 minutes, collect the supernatant, filter through a 0.22-micron filter membrane, and freeze-dry under vacuum at -40 to -30°C for 12-18 hours to obtain pomegranate peel-derived lactic acid bacteria lysate powder. Centrifugation and filtration remove broken bacterial fragments, ensuring the purity of the lysate; vacuum freeze-drying removes moisture at low temperatures, maximizing the preservation of active ingredients. Controlling the drying temperature and time within the above range ensures drying effectiveness and prevents product clumping. The content of the active ingredients of the lactic acid bacteria is detected by high-performance liquid chromatography (HPLC).

[0028] The preparation method of bamboo yellow mycelium extract is as follows: S3.1 Mycelial Culture: Take *Phyllostachys edulis* spawn and inoculate it into PDA medium (formula: 200g potato, 20g glucose, 15-20g agar, 1000ml purified water, sterilized at 121℃ for 20 minutes). Culture for 7-10 days at 25-28℃ and 60%-70% humidity to obtain *Phyllostachys edulis* mycelium. PDA medium is rich in nutrients, meeting the growth requirements of *Phyllostachys edulis* spawn. Maintaining the above-mentioned temperature and humidity promotes rapid growth of *Phyllostachys edulis* mycelium. A culture time of 7-10 days allows the mycelium to reach maturity and accumulate sufficient *Phyllostachys edulis* glycosides. This culture time was determined experimentally; mycelium matures in 7 days and begins to age after 10 days, hence this range was established.

[0029] S3.3 Pretreatment: The bamboo yellow mycelium is removed, rinsed 3-5 times with purified water, dried at 35℃ to constant weight, pulverized, and passed through an 80-mesh sieve to obtain bamboo yellow mycelium powder. Rinsing with purified water removes culture medium impurities from the mycelium surface; drying at 35℃ avoids high-temperature damage to bamboo red mycelium glycosides; and pulverizing and passing through an 80-mesh sieve increases the contact area between the mycelium and the extraction solvent, improving extraction efficiency.

[0030] S3.3 Extraction: Take 30-50 grams of bamboo yellow mycelium powder and add 400-600 ml of anhydrous ethanol, with a material-to-liquid ratio of 1:13 to 1:15. Extract at 30-35℃ for 20-24 hours, stirring every 4 hours at a speed of 100-120 rpm. Anhydrous ethanol, as the extraction solvent, can effectively extract active ingredients such as bamboo red mycelium glycosides from bamboo yellow mycelium. Controlling the material-to-liquid ratio, extraction temperature, and time within the above range ensures extraction efficiency while avoiding solvent waste and destruction of active ingredients. Stirring allows the mycelium powder to fully contact the solvent, improving extraction efficiency. This extraction time range was determined through parameter optimization experiments. When extracted for 20-24 hours, the extraction rate of bamboo red mycelium glycosides reaches over 85%; below 20 hours, the extraction rate is less than 70%; above 24 hours, the extraction rate does not significantly improve and increases solvent consumption.

[0031] S3.4 Purification: After extraction, centrifuge at 4000-5000 rpm for 10-12 minutes, collect the supernatant, and distill it under reduced pressure at 50-60℃ and 0.06-0.08 MPa to remove ethanol, obtaining a concentrated solution. Filter through a 0.22-micron filter membrane and freeze-dry under vacuum to obtain the bamboo yellow mycelium extract. Centrifugation removes undissolved mycelial residue, reduced pressure distillation removes ethanol at low temperature to avoid denaturation of bamboo yellow mycelium A due to high temperature, filtration further removes impurities, and freeze-drying preserves the activity of the active ingredients.

[0032] The preparation method of complex sophorolipids is as follows: S4.1, Strain Activation: Candida albicans was inoculated into activation medium (20-30 g glucose, 5-10 g yeast extract, 5-10 g peptone, 1000 ml purified water, sterilized at 121℃ for 20 minutes). The culture was then incubated with shaking at 28-32℃ and 150-180 rpm for 18-24 hours to obtain the activated strain. The activation medium is rich in nutrients, which can promote the activation and proliferation of Candida albicans. Shaking culture ensures sufficient contact between the strain and the medium, improving activation efficiency. Maintaining the culture temperature and rotation speed within the above range ensures the activity of the strain. The Candida albicans strain is deposited at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 2.538, and deposit date of January 10, 2025. The strain can be obtained directly from the depository institution.

[0033] S4.2 Fermentation Culture: Inoculate the activated strain into the fermentation medium at a volume of 5%-8% of the fermentation medium. The fermentation medium consists of 80-120g glucose, 50-80g coconut oil, 3-5g yeast extract, 1-2g potassium dihydrogen phosphate, 0.5-1g magnesium sulfate, and 1000ml purified water. Adjust the pH to 5.5-6.5 and ferment with shaking at 28-32℃ and 150-180 rpm for 48-72 hours. Glucose and coconut oil in the fermentation medium serve as carbon sources, promoting the synthesis of sophorolipids by Candida albicans. Potassium dihydrogen phosphate and magnesium sulfate act as nutrients, aiding in the growth of the strain. Adjusting the pH to 5.5-6.5 meets the growth requirements of Candida albicans. Shaking fermentation improves fermentation efficiency, ensuring the yield and quality of sophorolipids. The fermentation time range was determined through parameter optimization experiments. When fermenting for 48-72 hours, the yield of sophorolipids reaches 12-15 g / L. Below 48 hours, the yield is less than 8 g / L, and above 72 hours, the yield does not increase significantly and energy consumption increases.

[0034] S4.3 Separation and Purification: After fermentation, centrifuge the fermentation broth at 5000-6000 rpm for 10-15 minutes, collect the supernatant, adjust the pH to 2.0-2.5 with hydrochloric acid, let it stand for 2-3 hours to precipitate, collect the precipitate, wash it 2-3 times with anhydrous ethanol, and vacuum dry it at 40-50℃ for 8-12 hours to obtain the complex sophorolipid. Centrifugation removes bacterial cells and impurities; adjusting the pH to 2.0-2.5 causes the sophorolipid to precipitate; washing with anhydrous ethanol removes impurities from the precipitate; and vacuum drying removes moisture to obtain a high-purity complex sophorolipid.

[0035] The present invention also provides a method for preparing the above-mentioned compound formulation, comprising the following steps: S1: Weigh each component according to the specified mass fractions. Mix the kidney bean-honeysuckle fruit complex ferment, pomegranate peel-derived lactic acid bacteria lysate, bamboo fungus mycelium extract, and complex sophorolipids. Add 20-30 ml of purified water, stir well, and maintain a constant temperature of 30-35℃ for 30-40 minutes to obtain mixture A. Constant temperature stirring promotes the complete dissolution and mixing of the four components, ensuring their synergistic effect. Controlling the stirring time to 30-40 minutes avoids uneven mixing that could lead to decreased formulation stability.

[0036] S2: Add sodium hyaluronate, total asiaticosides, and glycerin to mixture A, and stir at 150-200 rpm for 20-30 minutes until completely dissolved to obtain mixture B. A higher stirring speed promotes rapid dissolution of sodium hyaluronate, total asiaticosides, and glycerin, preventing particle formation and ensuring the homogeneity of the formulation.

[0037] S3: Add Carbomer 940, trehalose, and chitosan, and continue stirring for 15-20 minutes. Adjust the stirring speed to 120-150 rpm and stir until the system is homogeneous and free of particles, obtaining mixture C. Reducing the stirring speed can prevent Carbomer 940 from clumping and ensure that Carbomer 940, trehalose, and chitosan are fully dispersed, so that the system reaches a homogeneous state.

[0038] S4: Add lactic acid and the remaining purified water, stir well, adjust the pH of the system to 3.8-4.5 with sodium hydroxide, add methylchloroisothiazolinone, and continue stirring for 10-15 minutes to obtain mixture D. Adjusting the pH to 3.8-4.5 matches the acidic environment of the vagina. Adding methylchloroisothiazolinone can extend the shelf life of the preparation, and stirring can ensure that methylchloroisothiazolinone is evenly dispersed.

[0039] S5: Sterilize the mixture D at 80-85℃ for 20-30 minutes, cool to room temperature, fill and seal to obtain the compound formulation. Controlling the sterilization temperature and time within the above range effectively kills bacteria in the mixture, ensuring the safety of the formulation; slow cooling avoids problems such as stratification and clumping due to sudden temperature changes; and aseptic filling avoids secondary contamination.

[0040] In step S5, the cooling rate to room temperature after sterilization is 2-3°C / minute. The filling environment is sterile. After filling, the product is sealed and stored at a temperature of 2-8°C. Slow cooling ensures the stability of the formulation, aseptic filling avoids contamination, and storage at 2-8°C maintains the activity of the active ingredients in the formulation and extends its shelf life.

[0041] This invention also provides the application of the above-mentioned compound preparation in the preparation of vaginal and pelvic floor muscle care products. This compound preparation can be formulated into various dosage forms such as gels, washes, and suppositories to meet the needs of different populations. It can effectively regulate the vaginal microecological balance, nourish the vaginal mucosa, enhance the toughness of pelvic floor muscle fibers, and promote the repair of pelvic floor muscle damage. It can be widely used in the field of feminine hygiene products.

[0042] Example 1: A compound preparation containing plant-derived fermented lactic acid bacteria lysate for nourishing and protecting the vaginal and pelvic floor muscles, comprising the following components by weight: Four parts of kidney bean-blue honeysuckle fruit complex fermentation product, five parts of pomegranate peel-derived lactic acid bacteria cell lysate, three parts of bamboo yellow mycelium extract, six parts of complex sophorolipid, two parts of sodium hyaluronate, one part of total glycosides of Centella asiatica, five parts of glycerol, one part of lactic acid, 36 parts of purified water, two parts of carbomer 940, one part of trehalose, one part of chitosan, 0.1 part of methylchloroisothiazolinone, and 0.1 part of sodium hydroxide.

[0043] Preparation of Kidney Bean-Honeysuckle Fruit Fermentation S1.1 Raw material pretreatment: Take 200g of fresh kidney beans and 150g of fresh honeysuckle berries. Remove impurities from the kidney beans, wash them and cut them into 1×1cm pieces. Remove the stems from the honeysuckle berries and wash them. Mix them together and put them into a juicer. Add 500ml of purified water and juice for 10 minutes to obtain kidney bean-honeysuckle berry mixed juice. S1.2 Sterilization treatment: Pour the mixed juice into the fermentation tank, adjust the initial pH value to 4.8, sterilize at 115℃ for 10 minutes, cool to 32℃, and obtain the fermentation substrate; S1.3 Inoculation and Fermentation: A compound lactic acid bacteria (Lactobacillus plantarum and Lactobacillus acidophilus in a 1:1 volume ratio) was inoculated into the fermentation substrate at 2% of the substrate volume. Fermentation was carried out at 34°C under anaerobic conditions for 24 hours, with stirring every 6 hours at a stirring speed of 120 rpm. The Lactobacillus plantarum was deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1295 and deposit date of January 10, 2025. The Lactobacillus acidophilus was deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1854 and deposit date of January 10, 2025. S1.4 Post-processing: After fermentation, centrifuge at 5000 rpm for 10 minutes, take the supernatant, filter through a 0.22 micron filter membrane to obtain the kidney bean-blue honeysuckle berry complex fermentation product, and store at 4℃ for later use. Preparation of pomegranate peel-derived lactic acid bacteria cell lysate S2.1 Preparation of pomegranate peel culture medium: Take 100g of dried pomegranate peel, crush it and pass it through an 80-mesh sieve, add 500ml of purified water, adjust the pH value to 3.0, the pomegranate peel mass concentration is 3g / ml, sterilize at 115℃ for 10 minutes, cool to 30℃, and obtain pomegranate peel fermentation culture medium. S2.2, Lactic acid bacteria proliferation: Inoculate the culture medium with Lactobacillus plantarum (preservation information is the same as in step S1.3), the inoculation amount is 4% of the culture medium volume, and incubate at 30℃ for 48 hours to achieve lactic acid bacteria proliferation; S2.3, Cell Isolation: After the proliferation is completed, filter to remove pomegranate peel residue, centrifuge at 5000 rpm for 5 minutes, take the precipitate, and obtain Lactobacillus plantarum cells; S2.4, Cell lysis treatment: Add the bacterial cells to 200 ml of physiological saline, stir evenly, and under ice bath conditions, use 250 watt ultrasonic waves to break them up, with a 1-second pause after every 2 seconds of operation, for a total of 25 minutes. S2.5 Purification: After the cell lysis is completed, the mixture is centrifuged at 5000 rpm for 10 minutes, the supernatant is collected, filtered through a 0.22-micron filter membrane, and freeze-dried at -40℃ for 12 hours to obtain the cell lysis product powder of pomegranate peel lactic acid bacteria. Preparation of bamboo yellow mycelium extract S3.1 Mycelial Culture: Take bamboo yellow fungus and inoculate it into PDA medium (formula: 200g potato, 20g glucose, 15g agar, 1000ml purified water, sterilized at 121℃ for 20 minutes). Culture for 7 days at 25℃ and 60% humidity to obtain bamboo yellow mycelium. S3.2 Pretreatment: Take out the mycelium, rinse it three times with purified water, dry it at 35℃ to constant weight, pulverize it and pass it through an 80-mesh sieve to obtain bamboo yellow mycelium powder; S3.3 Extraction: Take 30 g of powder, add 400 ml of anhydrous ethanol, with a material-to-liquid ratio of 1:13, and extract at 30°C for 20 hours, stirring once every 4 hours at a stirring speed of 100 rpm; S3.4 Purification: After extraction, centrifuge at 4000 rpm for 10 minutes, take the supernatant, remove ethanol by vacuum distillation at 50℃ and 0.06 MPa to obtain a concentrated solution, filter through a 0.22-micron filter membrane, and freeze dry under vacuum to obtain bamboo yellow mycelium extract. Preparation of complex sophorolipids S4.1, Strain Activation: Candida albicans was inoculated into activation medium (20 g glucose, 5 g yeast extract, 5 g peptone, 1000 ml purified water, sterilized at 121°C for 20 minutes), and cultured at 28°C with shaking at 150 rpm for 18 hours to obtain the activated strain; the Candida albicans strain is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 2.538 and deposit date of January 10, 2025; S4.2 Fermentation culture: Inoculate the activated strain into the fermentation medium (80 g glucose, 50 g coconut oil, 3 g yeast extract, 1 g potassium dihydrogen phosphate, 0.5 g magnesium sulfate, 1000 ml purified water), adjust the pH to 5.5, and ferment at 28℃ and 150 rpm for 48 hours. S4.3 Separation and purification: After fermentation, centrifuge at 5000 rpm for 10 minutes, take the supernatant, adjust the pH value to 2.0 with hydrochloric acid, let stand for 2 hours, precipitate, collect the precipitate, wash twice with anhydrous ethanol, and vacuum dry at 40℃ for 8 hours to obtain the complex sophorolipid. The preparation method of the above-mentioned compound preparation is as follows: S1: Weigh each component according to the mass fraction, mix the kidney bean-blue honeysuckle fruit complex ferment, pomegranate peel lactic acid bacteria cell lysate, bamboo yellow mycelium extract, and complex sophorolipid, add 20 ml of purified water, stir evenly, and stir at 30℃ for 30 minutes to obtain mixture A; S2: Add sodium hyaluronate, total asiaticoside, and glycerin, stir at 150 rpm for 20 minutes until completely dissolved to obtain mixture B; S3: Add carbomer 940, trehalose, and chitosan, and continue stirring for 15 minutes. Adjust the speed to 120 rpm and stir until the system is uniform and free of particles to obtain mixture C. S4: Add lactic acid and the remaining purified water (36-20=16 ml), stir well, adjust the pH of the system to 3.8 with sodium hydroxide, add methylchloroisothiazolinone, and continue stirring for 10 minutes to obtain mixture D; S5: Sterilize the mixture D at 80℃ for 20 minutes, cool it to room temperature at a rate of 2℃ / min, fill and seal it under aseptic conditions to obtain the compound preparation (gel), and store it at 2-8℃.

[0044] Example 2 (Preferred Example) A compound preparation containing plant-derived fermented lactic acid bacteria lysate for nourishing and protecting the vaginal and pelvic floor muscles, comprising the following components by weight: Six parts of kidney bean-blue honeysuckle fruit complex fermentation product, seven parts of pomegranate peel-derived lactic acid bacteria cell lysate, five parts of bamboo yellow mycelium extract, seven parts of complex sophorolipid, three and a half parts of sodium hyaluronate, two parts of total asiaticoside, seven and a half parts of glycerol, one and a half parts of lactic acid, four and a half parts of purified water, three and a half parts of carbomer 940, two and a half parts of trehalose, two and a half parts of chitosan, two and a half parts of methylchloroisothiazolinone, and one and a half parts of sodium hydroxide.

[0045] Preparation of Kidney Bean-Honeysuckle Fruit Fermentation S1.1 Raw material pretreatment: Take 250g of fresh kidney beans and 200g of fresh honeysuckle berries. Remove impurities from the kidney beans, wash them and cut them into 1×1cm pieces. Remove the stems from the honeysuckle berries and wash them. Mix them together and put them into a juicer. Add 600ml of purified water and juice for 12 minutes to obtain kidney bean-honeysuckle berry mixed juice. S1.2 Sterilization treatment: Pour the mixed juice into the fermentation tank, adjust the initial pH value to 5.0, sterilize at 118℃ for 12 minutes, cool to 34℃, and obtain the fermentation substrate; S1.3 Inoculation and Fermentation: A compound lactic acid bacteria (Lactobacillus plantarum and Lactobacillus acidophilus volume ratio 1:1) was inoculated into the fermentation substrate at a volume of 2.5% of the fermentation substrate volume. Fermentation was carried out at 35°C under anaerobic conditions for 27 hours, with stirring every 6 hours at a stirring speed of 135 rpm. The Lactobacillus plantarum was deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1295 and accession date of January 10, 2025. The Lactobacillus acidophilus was deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1854 and accession date of January 10, 2025. S1.4 Post-processing: After fermentation, centrifuge at 5500 rpm for 12 minutes, take the supernatant, filter through a 0.22 micron filter membrane to obtain the kidney bean-blue honeysuckle berry complex fermentation product, and store at 4℃ for later use. Preparation of pomegranate peel-derived lactic acid bacteria cell lysate S2.1 Preparation of pomegranate peel culture medium: Take 125g of dried pomegranate peel, crush it and pass it through a 90-mesh sieve, add 600ml of purified water, adjust the pH value to 3.5, the pomegranate peel mass concentration is 5g / ml, sterilize at 118℃ for 12 minutes, cool to 34℃, and obtain pomegranate peel fermentation culture medium. S2.2, Lactic acid bacteria proliferation: Inoculate the culture medium with Lactobacillus plantarum (preservation information is the same as in step S1.3), the inoculation amount is 6% of the culture medium volume, and incubate at 34℃ for 60 hours to achieve lactic acid bacteria proliferation; S2.3. Cell isolation: After the proliferation is completed, filter to remove pomegranate peel residue, centrifuge at 5500 rpm for 10 minutes, take the precipitate, and obtain Lactobacillus plantarum cells; S2.4, Cell lysis treatment: Add the bacterial cells to 250 ml of physiological saline, stir evenly, and under ice bath conditions, use 275 watt ultrasonic waves to break them up, with a 1-second pause after every 2 seconds of operation, for a total of 27 minutes. S2.5 Purification: After the cell lysis is completed, the mixture is centrifuged at 5500 rpm for 12 minutes, the supernatant is collected, filtered through a 0.22-micron filter membrane, and freeze-dried at -35℃ for 15 hours to obtain the cell lysis product powder of pomegranate peel-derived lactic acid bacteria. Preparation of bamboo yellow mycelium extract S3.1 Mycelial Culture: Take bamboo yellow fungus and inoculate it into PDA medium (formula: 200g potato, 20g glucose, 18g agar, 1000ml purified water, sterilized at 121℃ for 20 minutes), and culture it at 26.5℃ and 65% humidity for 8.5 days to obtain bamboo yellow mycelium; S3.2 Pretreatment: Take out the mycelium, rinse it 4 times with purified water, dry it at 35℃ to constant weight, pulverize it and pass it through an 80-mesh sieve to obtain bamboo yellow mycelium powder; S3.3 Extraction: Take 40 g of powder, add 500 ml of anhydrous ethanol, with a material-to-liquid ratio of 1:14, and extract at 32.5℃ for 22 hours, stirring once every 4 hours at a stirring speed of 110 rpm. S3.4 Purification: After extraction, centrifuge at 4500 rpm for 11 minutes, take the supernatant, remove ethanol by vacuum distillation at 55℃ and 0.07 MPa to obtain a concentrated solution, filter through a 0.22-micron filter membrane, and freeze dry under vacuum to obtain bamboo yellow mycelium extract. Preparation of complex sophorolipids S4.1, Strain Activation: Candida albicans was inoculated into activation medium (25 g glucose, 7.5 g yeast extract, 7.5 g peptone, 1000 ml purified water, sterilized at 121°C for 20 minutes), and cultured at 30°C with shaking at 165 rpm for 21 hours to obtain the activated strain; the Candida albicans strain is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 2.538 and deposit date of January 10, 2025; S4.2 Fermentation culture: Inoculate the activated strain into the fermentation medium (100 g glucose, 65 g coconut oil, 4 g yeast extract, 1.5 g potassium dihydrogen phosphate, 0.75 g magnesium sulfate, 1000 ml purified water), adjust the pH to 6.0, and ferment at 30℃ and 165 rpm for 60 hours with shaking. S4.3 Separation and Purification: After fermentation, centrifuge at 5500 rpm for 12 minutes, collect the supernatant, adjust the pH to 2.25 with hydrochloric acid, let stand for 2.5 hours, precipitate, collect the precipitate, wash twice with anhydrous ethanol, and vacuum dry at 45℃ for 10 hours to obtain the complex sophorolipid; the ratio of the acidic to lactone-type sophorolipids was detected by high performance liquid chromatography (HPLC), with methanol-water (volume ratio 70:30) as the mobile phase and 210 nm as the detection wavelength; The preparation method of the above-mentioned compound preparation is as follows: S1: Weigh each component according to the mass fraction, mix the kidney bean-blue honeysuckle fruit complex ferment, pomegranate peel-derived lactic acid bacteria cell lysate, bamboo yellow mycelium extract, and complex sophorolipid, add 25 ml of purified water, stir evenly, and stir at 32.5℃ for 35 minutes to obtain mixture A; S2: Add sodium hyaluronate, total asiaticosides, and glycerin, stir at 175 rpm for 25 minutes until completely dissolved to obtain mixture B; S3: Add carbomer 940, trehalose, and chitosan, and continue stirring for 17.5 minutes. Adjust the speed to 135 rpm and stir until the system is uniform and free of particles to obtain mixture C. S4: Add lactic acid and the remaining purified water (40-25=15 ml), stir well, adjust the pH of the system to 4.2 with sodium hydroxide, add methylchloroisothiazolinone, and continue stirring for 12.5 minutes to obtain mixture D; S5: Sterilize the mixture D at 82.5℃ for 25 minutes, cool it to room temperature at a rate of 2.5℃ / min, fill and seal it under aseptic conditions to obtain the compound preparation (gel), and store it at 2-8℃.

[0046] Example 3 A compound preparation containing plant-derived fermented lactic acid bacteria lysate for nourishing and protecting the vaginal and pelvic floor muscles, comprising the following components by weight: Eight parts of kidney bean-blue honeysuckle fruit complex fermentation product, nine parts of pomegranate peel-derived lactic acid bacteria cell lysate, seven parts of bamboo yellow mycelium extract, eight parts of complex sophorolipid, five parts of sodium hyaluronate, three parts of total asiaticoside, ten parts of glycerol, two parts of lactic acid, four and five parts of purified water, four parts of carbomer 940, three parts of trehalose, three parts of chitosan, three parts of methylchloroisothiazolinone, and two and two parts of sodium hydroxide.

[0047] Preparation of Kidney Bean-Honeysuckle Fruit Fermentation S1.1 Raw material pretreatment: Take 300g of fresh kidney beans and 250g of fresh honeysuckle berries. Remove impurities from the kidney beans, wash them and cut them into 1×1cm pieces. Remove the stems from the honeysuckle berries and wash them. Mix them together and put them into a juicer. Add 700ml of purified water and juice for 15 minutes to obtain kidney bean-honeysuckle berry mixed juice. S1.2 Sterilization treatment: Pour the mixed juice into the fermentation tank, adjust the initial pH value to 5.2, sterilize at 120℃ for 15 minutes, cool to 36℃, and obtain the fermentation substrate; S1.3 Inoculation and Fermentation: A compound lactic acid bacteria (Lactobacillus plantarum and Lactobacillus acidophilus volume ratio 1:1) was inoculated into the fermentation substrate at a volume of 3% of the substrate volume. Fermentation was carried out at 36°C under anaerobic conditions for 30 hours, with stirring every 6 hours at a stirring speed of 150 rpm. The Lactobacillus plantarum was deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1295 and deposit date of January 10, 2025. The Lactobacillus acidophilus was deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1854 and deposit date of January 10, 2025. S1.4 Post-processing: After fermentation, centrifuge at 6000 rpm for 15 minutes, take the supernatant, filter through a 0.22 micron filter membrane to obtain the kidney bean-honeysuckle fruit complex fermentation product, and store at 4℃ for later use. Preparation of pomegranate peel-derived lactic acid bacteria cell lysate S2.1 Preparation of pomegranate peel culture medium: Take 150g of dried pomegranate peel, crush it and pass it through a 100-mesh sieve, add 700ml of purified water, adjust the pH value to 4.0, the pomegranate peel mass concentration is 7g / ml, sterilize at 120℃ for 15 minutes, cool to 37℃, and obtain pomegranate peel fermentation culture medium. S2.2, Lactic acid bacteria proliferation: Inoculate the culture medium with Lactobacillus plantarum (preservation information is the same as in step S1.3), the inoculation amount is 8% of the culture medium volume, and incubate at 37℃ for 72 hours to achieve lactic acid bacteria proliferation; S2.3. Cell isolation: After the proliferation is completed, filter to remove pomegranate peel residue, centrifuge at 6000 rpm for 15 minutes, take the precipitate, and obtain Lactobacillus plantarum cells; S2.4, Cell lysis treatment: Add the bacterial cells to 300 ml of physiological saline, stir evenly, and under ice bath conditions, use 300 watt ultrasonic waves to break them up, with a 1-second pause after every 2 seconds of operation, for 30 minutes. 2.5 Purification: After lysing, the sample was centrifuged at 6000 rpm for 15 minutes, the supernatant was collected, filtered through a 0.22-micron filter membrane, and freeze-dried at -30℃ for 18 hours to obtain the lysate powder of pomegranate peel-derived lactic acid bacteria. Preparation of bamboo yellow mycelium extract S3.1 Mycelial Culture: Take bamboo yellow fungus and inoculate it into PDA medium (formula: 200g potato, 20g glucose, 20g agar, 1000ml purified water, sterilized at 121℃ for 20 minutes). Culture it at 28℃ and 70% humidity for 10 days to obtain bamboo yellow mycelium. S3.2 Pretreatment: Take out the mycelium, rinse it 5 times with purified water, dry it at 35℃ to constant weight, pulverize it and pass it through an 80-mesh sieve to obtain bamboo yellow mycelium powder; S3.3 Extraction: Take 50 g of powder, add 600 ml of anhydrous ethanol, with a material-to-liquid ratio of 1:15, and extract at 35°C for 24 hours, stirring once every 4 hours at a stirring speed of 120 rpm. S3.4 Purification: After extraction, centrifuge at 5000 rpm for 12 minutes, take the supernatant, remove ethanol by vacuum distillation at 60℃ and 0.08 MPa to obtain a concentrated solution, filter through a 0.22-micron filter membrane, and freeze dry under vacuum to obtain bamboo yellow mycelium extract. Preparation of complex sophorolipids S4.1, Strain Activation: Candida albicans was inoculated into activation medium (30 g glucose, 10 g yeast extract, 10 g peptone, 1000 ml purified water, sterilized at 121°C for 20 minutes), and cultured at 32°C with shaking at 180 rpm for 24 hours to obtain the activated strain; the Candida albicans strain is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 2.538 and deposit date of January 10, 2025; S4.2 Fermentation culture: Inoculate the activated strain into the fermentation medium (120 g glucose, 80 g coconut oil, 5 g yeast extract, 2 g potassium dihydrogen phosphate, 1 g magnesium sulfate, 1000 ml purified water), adjust the pH to 6.5, and ferment at 32℃ and 180 rpm for 72 hours. S4.3 Separation and purification: After fermentation, centrifuge at 6000 rpm for 15 minutes, take the supernatant, adjust the pH value to 2.5 with hydrochloric acid, let stand for 3 hours, precipitate, collect the precipitate, wash 3 times with anhydrous ethanol, and vacuum dry at 50℃ for 12 hours to obtain the complex sophorolipid. The preparation method of the above-mentioned compound preparation is as follows: S1: Weigh each component according to the mass fraction, mix the kidney bean-blue honeysuckle fruit complex ferment, pomegranate peel lactic acid bacteria cell lysate, bamboo yellow mycelium extract, and complex sophorolipid, add 30 ml of purified water, stir evenly, and stir at 35℃ for 40 minutes to obtain mixture A. S2: Add sodium hyaluronate, total asiaticoside, and glycerin, stir at 200 rpm for 30 minutes until completely dissolved to obtain mixture B; S3: Add carbomer 940, trehalose, and chitosan, and continue stirring for 20 minutes. Adjust the speed to 150 rpm and stir until the system is uniform and free of particles to obtain mixture C. S4: Add lactic acid and the remaining purified water (45-30=15 ml), stir well, adjust the pH of the system to 4.5 with sodium hydroxide, add methylchloroisothiazolinone, and continue stirring for 15 minutes to obtain mixture D; S5: Sterilize the mixture D at 85℃ for 30 minutes, cool it to room temperature at a rate of 3℃ / min, fill and seal it under aseptic conditions to obtain the compound preparation (gel), and store it at 2-8℃.

[0048] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0049] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A compound preparation of plant-fermented lactic acid bacteria lysate for nourishing and protecting the vaginal and pelvic floor muscles, characterized in that, By weight, it includes the following components: Kidney bean-honeysuckle berry fermented compound: 4-8 parts; Pomegranate peel-derived lactic acid bacteria lysate: 5-9 parts; Bamboo fungus mycelium extract: 3-7 parts; Complex sophorolipids: 6-8 parts; Sodium hyaluronate: 2-5 parts; Total glycosides from Centella asiatica: 1-3 parts; Glycerin: 5-10 parts; Lactic acid: 1-2 parts; Purified water: 36-45 parts; Carbomer 940: 2-4 servings; Trehalose: 1-3 parts; Chitosan: 1-3 parts; Methylchloroisothiazolinone: 0.1-0.3 parts; Sodium hydroxide: 0.1-0.2 parts.

2. The compound preparation of plant-fermented lactic acid bacteria lysate for nourishing and protecting the vaginal and pelvic floor muscles according to claim 1, characterized in that, The preparation method of the kidney bean-honeysuckle fruit complex ferment is as follows: S1.1 Raw material pretreatment: Take 200-300 grams of fresh kidney beans and 150-250 grams of fresh honeysuckle berries. Remove impurities from the kidney beans, wash them, and cut them into 1×1 cm pieces. Remove the stems from the honeysuckle berries, wash them, mix the two together, put them into a juicer, add 500-700 ml of purified water, and juice for 10-15 minutes to obtain kidney bean-honeysuckle berry mixed juice. S1.2 Sterilization treatment: Pour the mixed juice into the fermentation tank, adjust the initial pH value to 4.8-5.2, sterilize at 115-120℃ for 10-15 minutes, cool to 32-36℃ to obtain the fermentation substrate; S1.3 Inoculation and Fermentation: Inoculate the fermentation substrate with a compound lactic acid bacteria at a volume of 2%-3% of the substrate. The compound lactic acid bacteria is a mixture of *Lactobacillus plantarum* and *Lactobacillus acidophilus*, with a volume ratio of 1:

1. Ferment at 34-36℃ under anaerobic conditions for 24-30 hours, stirring every 6 hours at a speed of 120-150 rpm. The *Lactobacillus plantarum* is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 1.1295 and a deposit date of January 10, 2025. The *Lactobacillus acidophilus* is deposited at the same center with accession number CGMCC No. 1.1854 and a deposit date of January 10, 2025. S1.4 Post-processing: After fermentation, centrifuge the fermentation broth at 5000-6000 rpm for 10-15 minutes, take the supernatant, filter it through a 0.22-micron filter membrane to obtain the kidney bean-blue honeysuckle berry complex fermentation product, and store it at 4℃ for later use.

3. The compound preparation of plant-fermented lactic acid bacteria lysate for nourishing and protecting the vaginal pelvic floor muscles according to claim 1, characterized in that, The preparation method of the pomegranate peel-derived lactic acid bacteria lysate is as follows: S2.1 Preparation of pomegranate peel culture medium: Take 100-150 grams of dried pomegranate peel, crush it and pass it through an 80-100 mesh sieve, add 500-700 ml of purified water, adjust the pH value to 3.0-4.0, the pomegranate peel mass concentration is 3-7 g / ml, sterilize at 115-120℃ for 10-15 minutes, cool to 30-37℃, and obtain pomegranate peel fermentation culture medium; S2.2, Lactic acid bacteria proliferation: Inoculate pomegranate peel fermentation medium with Lactobacillus plantarum at a volume of 4%-8% of the medium volume, and culture at 30-37℃ for 48-72 hours to achieve lactic acid bacteria proliferation; S2.

3. Cell isolation: After the proliferation is completed, first filter to remove pomegranate peel residue, then centrifuge the remaining liquid at a speed of 5000-6000 rpm for 5-15 minutes, take the precipitate, and obtain Lactobacillus plantarum cells; S2.4, Cell lysis treatment: Add Lactobacillus plantarum cells to 200-300 ml of physiological saline, stir well, place in an ice bath environment, and use ultrasonic disruption method to perform cell lysis. The ultrasonic power is 250-300 watts, with a 1-second interval between every 2 seconds of operation, and the disruption time is 25-30 minutes. S2.5 Purification: After lysing, centrifuge at 5000-6000 rpm for 10-15 minutes, take the supernatant, filter through a 0.22-micron filter membrane, freeze dry under vacuum at -40 to -30℃ for 12-18 hours to obtain pomegranate peel-derived lactic acid bacteria lysate powder.

4. The compound preparation of plant-fermented lactic acid bacteria lysate for nourishing and protecting the vaginal pelvic floor muscles according to claim 1, characterized in that, The preparation method of the bamboo yellow mycelium extract is as follows: S3.1 Mycelial Culture: Take bamboo yellow fungus, inoculate it into PDA medium, and culture it for 7-10 days at 25-28℃ and 60%-70% humidity to obtain bamboo yellow mycelium; PDA medium formula: 200g potato, 20g glucose, 15-20g agar, 1000ml purified water, sterilize at 121℃ for 20 minutes; S3.2 Pretreatment: Take out the bamboo yellow mycelium, rinse it with purified water 3-5 times, dry it at 35℃ to constant weight, pulverize it and pass it through an 80-mesh sieve to obtain bamboo yellow mycelium powder; S3.3 Extraction: Take 30-50 grams of bamboo yellow mycelium powder, add 400-600 ml of anhydrous ethanol, with a material-to-liquid ratio of 1:13 to 1:15, and extract at 30-35℃ for 20-24 hours, stirring once every 4 hours during the process, with a stirring speed of 100-120 rpm. S3.4 Purification: After extraction, centrifuge at 4000-5000 rpm for 10-12 minutes, collect the supernatant, and distill the supernatant under reduced pressure at 50-60℃ and 0.06-0.08 MPa to remove ethanol, obtain a concentrated solution, filter through a 0.22-micron filter membrane, and freeze-dry under vacuum to obtain bamboo yellow mycelium extract.

5. The compound preparation of plant-fermented lactic acid bacteria lysate for nourishing and protecting the vaginal and pelvic floor muscles according to claim 1, characterized in that, The preparation method of the complex sophorolipid is as follows: S4.1, Strain Activation: Candida albicans was inoculated into an activation medium and cultured with shaking at 28-32℃ and 150-180 rpm for 18-24 hours to obtain the activated strain. The activation medium formula was: 20-30 g glucose, 5-10 g yeast extract, 5-10 g peptone, and 1000 ml purified water, sterilized at 121℃ for 20 minutes. The Candida albicans strain was deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 2.538 and accession date January 10, 2025. S4.2 Fermentation culture: Inoculate the activated strain into the fermentation medium at a volume of 5%-8% of the fermentation medium. The fermentation medium consists of 80-120g glucose, 50-80g coconut oil, 3-5g yeast extract, 1-2g potassium dihydrogen phosphate, 0.5-1g magnesium sulfate, and 1000ml purified water. Adjust the pH to 5.5-6.5 and ferment with shaking at 28-32℃ and 150-180 rpm for 48-72 hours. S4.3 Separation and purification: After fermentation, centrifuge the fermentation broth at 5000-6000 rpm for 10-15 minutes, take the supernatant, adjust the pH value to 2.0-2.5 with hydrochloric acid, let it stand for 2-3 hours, precipitate, collect the precipitate, wash it 2-3 times with anhydrous ethanol, and vacuum dry it at 40-50℃ for 8-12 hours to obtain the complex sophorolipid.

6. The preparation method of the compound preparation of plant fermented lactic acid bacteria lysate for nourishing and protecting the vaginal pelvic floor muscles according to claim 1, characterized in that, Includes the following steps: S1: Weigh each component according to the mass fraction, mix the kidney bean-blue honeysuckle fruit complex ferment, pomegranate peel-derived lactic acid bacteria cell lysate, bamboo yellow mycelium extract, and complex sophorolipid, add 20-30 ml of purified water, stir evenly, and stir at a constant temperature of 30-35℃ for 30-40 minutes to obtain mixture A; S2: Add sodium hyaluronate, total asiaticoside, and glycerin to mixture A, stir at 150-200 rpm for 20-30 minutes until completely dissolved to obtain mixture B; S3: Add carbomer 940, trehalose, and chitosan, and continue stirring for 15-20 minutes. Adjust the stirring speed to 120-150 rpm and stir until the system is uniform and free of particles to obtain mixture C. S4: Add lactic acid and the remaining purified water, stir well, adjust the pH of the system to 3.8-4.5 with sodium hydroxide, add methylchloroisothiazolinone, and continue stirring for 10-15 minutes to obtain mixture D; S5: Sterilize the mixture D at 80-85℃ for 20-30 minutes, cool to room temperature, fill and seal to obtain the compound preparation.

7. The preparation method of the compound preparation of plant fermented lactic acid bacteria cell lysate for nourishing and protecting the vaginal pelvic floor muscles according to claim 6, characterized in that, In step S5, the rate of cooling to room temperature after sterilization is 2-3℃ / minute, the filling environment is aseptic, and the contents are sealed and stored at a temperature of 2-8℃ after filling.

8. The application of the compound preparation according to claim 1 in the preparation of vaginal and pelvic floor muscle care products.