Pediococcus acidilactici NH3 for treating male prostate and improving sexual function as well as culture method and application thereof

Abstract

The invention relates to the technical field of probiotics, in particular to pediococcus acidilactici NH3 for treating male prostate and improving sexual function as well as a culture method and application of the pediococcus acidilactici NH3. The strain is preserved in Guangdong Microbial Culture Collection Center on April 18, 2025, and the preservation number is GDMCC No: 66163. The strain is a probiotic strain approved by the National Health Commission. The strain has the effects of regulating intestinal flora balance and promoting digestion and absorption of a human body, and also has the effects of treating male prostate and improving the sexual function. 5 billion strains and 50 milligrams of metabolites are given every day, so that the endophytic essential quantity of astragaloside, spinosin and psoralen can be continuously provided in the intestinal reproduction and growth process, the male prostate can be treated, the sexual function can be improved, and the body health can be kept.

Description

Technical Field

[0001] This invention relates to the field of probiotics technology, and in particular to a type of Pyrococcus lactis NH3 for treating male prostate problems and improving sexual function, as well as its culture method and application. Background Technology

[0002] The information disclosed in the background section of this invention is intended only to enhance the understanding of the overall background of the invention and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] Statistics show that the incidence of chronic prostatitis in my country ranges from 6.0% to 32.9%, and the prevalence of erectile dysfunction in adult men reaches 49.69%. Furthermore, the incidence of benign prostatic hyperplasia (BPH) is approximately 50% in men over 50 years of age, and as high as 90% in those over 80 years of age. These diseases pose a significant threat to men's physical and mental health and quality of life. In recent years, the role of the gut microbiota in human health has received increasing attention, with numerous studies revealing its association with various diseases such as rheumatoid arthritis, gastrointestinal diseases, cardiovascular diseases, and Alzheimer's disease. Although no direct effect of the gut microbiota on the prostate has yet been found, there is evidence suggesting that it may affect prostate health through indirect pathological mechanisms.

[0004] Studies have shown that a high-fat diet can induce gut microbiota dysbiosis in mice and significantly reduce the expression of tight junction proteins in the colonic epithelium. When the gut microbiota is disordered or imbalanced, the intestinal barrier function may be impaired, leading to increased intestinal permeability and triggering "leaky gut syndrome." The translocation of gut microbiota and its metabolites from the gut activates the immune system, leading to inflammation and the release of pro-inflammatory factors. If gut microbiota metabolites (such as short-chain fatty acids (SCFAs)) or bacterial components (such as lipopolysaccharides) enter the systemic circulation via venous and lymphatic pathways, they may induce a systemic inflammatory response and cause prostatitis.

[0005] The gut microbiota produces a variety of bioactive metabolites, such as bile acid derivatives, SCFAs, and tryptophan metabolites. These substances participate in the regulation of host physiological and pathological processes through different mechanisms and pathways. SCFAs are among the most important metabolites in the gut microbiota, mainly produced by the fermentation and decomposition of dietary fiber by gut microbiota, including acetic acid, propionic acid, and butyric acid. SCFAs are crucial for regulating the immune system, metabolism, and cell proliferation. SCFAs are associated with urinary system diseases such as prostate cancer (PCa), bladder cancer, and benign prostatic hyperplasia. SCFAs are recognized by G protein-coupled receptors (GPCRs); G protein-coupled receptor 43 (GPR43), G protein-coupled receptor 41 (GPR41), and G protein-coupled receptor 109A (GPR109A) have been identified as SCFA receptors. SCFA can bind to GPR43 and regulate various immune cells, including regulatory T cells (Treg cells), G protein-coupled receptor 109A (Th17 cells), macrophages, and B cells, by inhibiting histone deacetylase (HDAC), thereby suppressing inflammation. Summary of the Invention

[0006] In view of this, the present invention provides a *Pediococcus lactis* NH3 strain for treating male prostate problems and improving sexual function, along with its culture method and application. This bacterium was isolated from fresh oyster meat and identified as *Pediococcus lactis* using the 16S rRNA method. This strain, while possessing the functions of regulating intestinal flora balance and promoting human digestion and absorption, also has the effect of treating male prostate problems and improving sexual function.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solution: In a first aspect, the present invention provides a *Pediococcus lactis* NH3 strain for treating male prostate problems and improving sexual function. Pediococcus acidilactici This strain was deposited on April 18, 2025, at the Guangdong Provincial Center for Microbial Culture Collection (Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou), with accession number GDMCC No: 66163.

[0008] Secondly, the present invention provides the above-mentioned *Pediococcus lactis* NH3 ( Pediococcus acidilactici The cultivation method includes: scale-up culture, primary seed culture, seed tank culture and fermentation culture.

[0009] Furthermore, the culture medium for the expanded culture contains, per 1000 mL of water: 15-25 g lactose, 3.25-3.75 g raffinose, 4-6 g inulin, 2-5 g glucosyl alcohol, 0.5-2 g sodium acetate, 0.5-1 mL Tween-80, and 0.6-1 g dipotassium hydrogen phosphate; the pH of the culture medium for the expanded culture is adjusted to 6.0-6.5; preferably, the culture conditions for the expanded culture are static culture at 36-38℃ for 10-20 h.

[0010] Furthermore, the culture medium for the expanded culture contains, per 1000 mL of water: 20.0 g lactose, 3.50 g raffinose, 5.0 g inulin, 3.0 g g glucose alcohol, 1.0 g sodium acetate, 0.5 mL Tween-80, and 0.8 g dipotassium hydrogen phosphate; the pH of the culture medium for the expanded culture is adjusted to 6.0; preferably, the culture conditions for the expanded culture are static culture at 37°C for 16 h.

[0011] Furthermore, the culture medium for the primary seed culture contains, per 1000 mL of water: 2-5 g lactose, 1-3 g glucose alcohol, 4-6 g casein peptone, 1-3 g inulin, 2-4 g beef extract, 1-3 g arginine, 0.5-2 g sodium acetate, 0.5-2 g threonine, and 0.5-1 mL Tween-80; the pH of the culture medium for the primary seed culture is natural; preferably, the culture conditions for the primary seed culture are 36-38℃ for 20-30 h; the inoculum size for the primary seed culture is 0.4-0.6% by volume.

[0012] Furthermore, the culture medium for the primary seed culture contains, per 1000 mL of water: 3 g lactose, 2 g glucose alcohol, 5 g casein peptone, 2 g inulin, 3 g beef extract powder, 2 g arginine, 1 g sodium acetate, 1 g threonine, and 1 mL Tween-80; the pH of the culture medium for the primary seed culture is natural; preferably, the culture conditions for the primary seed culture are culture at 37°C for 25 h; the inoculum amount for the primary seed culture is 0.5% by volume.

[0013] Further, the culture medium for the seed tank culture comprises, by weight percentage: lactose 2-4%, glucose 0.5-1.5%, dextran 0.5-1%, casein peptone 0.3-0.6%, potassium dihydrogen phosphate 0.2-0.5%, sodium acetate 0.6-1.0%, arginine 0.1-0.3%, Tween-80 0.1-0.2%, polyether defoamer 0.05-0.15%, and tap water 92-94%; the pH of the culture medium for the seed tank culture is natural; preferably, the culture conditions for the seed tank culture are: tank pressure 0.04-0.06 MPa, culture at 36-38℃ for 10-20 h; the inoculum size for the seed tank culture is 4-6%, by volume.

[0014] Further, the culture medium for the seed tank culture comprises, by weight percentage: 3% lactose, 1% glucan, 0.5% dextran, 0.4% casein peptone, 0.3% potassium dihydrogen phosphate, 0.8% sodium acetate, 0.2% arginine, 0.1% Tween-80, 0.1% polyether defoamer, and 93.6% tap water; the pH of the culture medium for the seed tank culture is natural. Preferably, the culture conditions for the seed tank culture are: tank pressure 0.05 MPa, culture at 37°C for 15 h; the inoculum size for the seed tank culture is 5% by volume.

[0015] Further, the fermentation culture medium comprises, by weight percentage: lactose 3-5%, casein peptone 0.6-1.0%, dextran 0.5-1.5%, raffinose 1.5-2.0%, yeast extract 1.0-1.5%, sodium acetate 0.2-0.5%, Tween-80 0.1-0.3%, soybean protein powder 0.5-1.5%, dipotassium hydrogen phosphate 0.8-1.0%, polyether defoamer 0.05-0.15%, and tap water 87-89%; preferably, the pH value of the fermentation culture medium is 6.0-7.0; preferably, the fermentation culture conditions are: tank pressure 0.04-0.06 MPa, temperature 36-38℃, and time 10-20 h.

[0016] Further, the fermentation culture medium comprises, by weight percentage: 4% lactose, 0.8% casein peptone, 1% dextran, 1.7% raffinose, 1.2% yeast extract, 0.3% sodium acetate, 0.2% Tween-80, 1% soybean protein powder, 0.9% dipotassium hydrogen phosphate, 0.1% polyether defoamer, and 88.8% tap water; preferably, the pH value of the fermentation culture medium is 6.5; preferably, the fermentation culture conditions are: tank pressure 0.05 MPa, temperature 37°C, and time 15 h.

[0017] Thirdly, the present invention provides a microbial inoculant comprising an active ingredient, said active ingredient comprising the *Pediococcus lactis* NH3 or its fermentation broth as described in the first aspect.

[0018] Furthermore, the microbial agent comprises inactivated Pediococcus lactis NH3 or its fermentation broth.

[0019] Fourthly, the present invention provides the use of the *Pediococcus lactis* NH3 described in the first aspect or the microbial agent described in the third aspect in the preparation of health foods that help regulate intestinal flora or aid digestion.

[0020] Fifthly, the present invention provides the use of the *Pediococcus lactis* NH3 described in the first aspect or the microbial agent described in the third aspect in the preparation of a medicament for treating nonbacterial prostatitis, benign prostatic hyperplasia, and kidney deficiency.

[0021] Furthermore, the drug exists in the form of lyophilized bacterial powder.

[0022] Sixthly, the present invention provides a method for preparing freeze-dried bacterial powder for treating nonbacterial prostatitis, benign prostatic hyperplasia, and kidney deficiency, comprising: The fermentation broth obtained after the second fermentation culture is inactivated and centrifuged to obtain the final product; the metabolites of the inactivated wet bacterial mud are freeze-dried, then mixed with auxiliary materials to prepare a suspension, and the suspension is freeze-dried to obtain the final product.

[0023] Furthermore, the inactivation refers to UHT inactivation treatment of the fermentation broth at 100~140℃ for 5~8s.

[0024] Furthermore, the centrifugation speed is 8000~12000 r / min; specifically, centrifugation is performed at a feed rate of 1~3 kg / min.

[0025] Furthermore, the moisture content of the freeze-dried bacterial powder is ≤ 4% by mass.

[0026] Furthermore, the excipients include lactose, astragalus powder, jujube seed powder, and fucoidan.

[0027] Furthermore, the mass ratio of the inactivated wet bacterial sludge metabolites to the auxiliary materials is 1:2~2.5.

[0028] Compared with the prior art, the present invention has achieved the following beneficial effects: (1) The Lactococcus lactis NH3 provided by the present invention has the functions of regulating the balance of intestinal flora and promoting human digestion and absorption, as well as treating male prostate and improving sexual function. It has the characteristics of being non-toxic, effective, safe and green, and can play an important role in human health.

[0029] (2) The finished product made from the lactic acid cocci NH3 provided by the present invention has a significant effect on the treatment of non-bacterial prostatitis, benign prostatic hyperplasia and kidney deficiency. Administering 5 billion strains of the present invention and 50 mg of its metabolites daily can sustainably provide the necessary endogenous amounts of astragaloside A, spinosin, and psoralen during the intestinal reproduction and growth process, which can treat male prostatitis, improve sexual function, and maintain the body's health. Attached Figure Description

[0030] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0031] Figure 1 This is a comparative graph showing the levels of TNF-α, IL-8, and IL-10 in prostate tissue homogenates from different groups in this invention. Figure 2 This is a graph showing the effect of the present invention on the prostate index in mice; Figure 3 This is a graph showing the effect of the present invention on mouse serum T levels; Figure 4 This is a graph showing the effect of the present invention on serum DHT and E2 levels in mice; Figure 5 This is a diagram showing the effect of the present invention on the capture latency of mice; Figure 6 This is a diagram showing the effect of the present invention on the mounting latency and ejaculation latency in mice; Figure 7 This is a graph showing the effect of the present invention on the number of mouse captures, mountings, and ejaculations; Figure 8 This is a graph showing the effect of the present invention on serum Cr concentration; Figure 9 This is a graph showing the effect of the present invention on serum BUN concentration; Figure 10 This is a diagram showing the effect of the present invention on LH, FSH, and E2 levels; Figure 11 This is a diagram showing the effect of the present invention on GnRH levels; Figure 12 This is a graph showing the effect of the present invention on T levels; Figure 13 This is a diagram showing the effect of the present invention on ACP in the prostate and fructose in the seminal vesicle of mice; Figure 14 This is a graph showing the effect of the present invention on NO and cGMP in the mouse penis; Figure 15 This is a diagram showing the effect of the present invention on NOS in the mouse penis; Figure 16 This is a diagram showing the effect of the present invention on PDE-5 in the mouse penis. Detailed Implementation

[0032] It should be noted that the following detailed descriptions are exemplary and intended to provide further illustration of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0033] The technical solution of the present invention will be further described below with reference to specific embodiments.

[0034] Example 1: Strain Identification A strain of *Pediococcus lactis* was isolated from fresh oyster meat and identified as *Pediococcus lactis* NH3 by the 16S rRNA method. The 16S rRNA sequence of the strain is shown in SEQ ID NO:1.

[0035] The *Pediococcus lactis* NH3 strain screened above was deposited on April 18, 2025, at the Guangdong Provincial Center for Microbial Culture Collection (Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou), with accession number GDMCC No: 66163.

[0036] SEQ ID NO:1 GGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCGTGTAGGCGAGTTGCAGCCTACAGTCCGAACTGAGAATGGTTTTAAGAGATTAGCTAAACCTCGCGGTTTCGCGACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTC GTCCCCACCTTCCTCCCGGTTTGTCACCGGCAGTCTCACTAGAGTGCCCAACTGAATGCTGGCAACTAGTAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCATTCTGTCCCCGAAGGGAACACCTAATCTCTTA GGTTGGCAGAAGATGTCAAGACCTGGTAAGGTTCTTCGCGTAGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGCCCCGTCAATTCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGATTACTTAATGCGTTAGCTGCAGCACTGAAGGGCGGAAACCCTCCAAC ACTTAGTAATCATCGTTTACGGCATGGACTACCAGGGTATCTAATCCTGTTCGCTACCCATGCTTTCGAGCCTCAGCGTCAGTTACAGACCAGACAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACTCA Example 2: Culture of the strain Equipment used in production: microbial culture chamber, clean bench, constant temperature incubator, various test tubes, Erlenmeyer flasks, seed tanks, fermentation tanks, vacuum freeze dryer, three-dimensional mixer, low temperature pulverizer, vacuum packaging machine.

[0037] I. Expanding the culture of microbial strains Microbial culture: Large-scale culture of microbial strains stored in a -80℃ refrigerator.

[0038] Culture medium preparation: lactose 20.0g, raffinose 3.50g, inulin 5.0g, glucol 3.0g, sodium acetate 1.0g, Tween-80 0.5mL, dipotassium hydrogen phosphate 0.8g, tap water 1000mL, pH adjusted to 6.0.

[0039] Procedure: Add the lactose and other ingredients from the above formula to 1000mL of water in the formula without any order of addition, stir for 10 minutes to dissolve, then dispense 5mL into test tubes, seal each test tube with kraft paper stoppers, and sterilize in a medical sterilizer at 0.1~0.12MPa for 20 minutes. The culture medium is now ready.

[0040] Inoculation: Under aseptic conditions (clean bench), open the cryovial containing bacteria stored at -80℃ and thaw it at room temperature. Then, take one vial of sterilized culture medium, add 0.5 mL of the thawed bacterial solution to the vial, seal the vial with kraft paper and gently shake it. Place it in a constant temperature incubator at 37℃ for 16 hours. This is called the expanded culture medium.

[0041] II. Primary Seed Culture Primary seed culture formula: lactose 3g, glucol 2g, casein peptone 5g, inulin 2g, beef extract powder 3g, arginine 2g, sodium acetate 1g, threonine 1g, Tween-80 1mL, tap water 1000mL, pH value natural.

[0042] Procedure: Add the above raw materials to 1000mL of tap water without regard to the order of addition, turn on the stirrer and stir for 10 minutes until completely dissolved. This is called the first-stage seed culture medium.

[0043] Add the prepared primary seed culture medium, after stirring and dissolving, to the Erlenmeyer flask according to the required dosage. Then, wrap the flask opening with six layers of gauze and one layer of kraft paper. Place the flask in a sterilizer for sterilization. Before sterilization, first open the exhaust valve on the sterilizer until a small amount of steam is released, then close it. When the pressure on the sterilizer reaches 0.05 MPa, open the exhaust valve again for 6-8 minutes to release the steam. Then close the exhaust valve and continue heating. Start timing when the steam pressure in the sterilizer reaches 0.1-0.12 MPa. The sterilization time is 20 minutes. After sterilization, remove the flask from the heat source and allow it to cool naturally. When the pressure gauge on the sterilizer reaches zero, turn on the sterilizer, take out the sterilized culture medium and place it in the laminar flow hood to cool naturally. When the temperature drops to 37℃, transfer the above-cultured expansion inoculum to an Erlenmeyer flask containing sterilized culture medium. The inoculation volume is 0.5%, that is, 100mL of primary seed culture medium and 0.5mL of expansion inoculum. After completion, seal the bottle mouth with the original gauze and kraft paper, and shake it slightly by hand to mix the inoculated inoculum with the culture medium evenly. Then place it in a constant temperature incubator and incubate at 37℃ for 25 hours. This is called primary inoculum.

[0044] III. Seed Tank Cultivation Sterilization of fermentation equipment, pipelines, and aseptic filtration systems: First, open the valves of all inlet and outlet pipes and the sterile air pipe, and introduce steam at 0.12-0.14 MPa to allow the steam to connect with the pipe valves and a small amount of steam to be discharged. Continuous ventilation for 40 minutes, then close the valves of all inlet and outlet pipes and set aside for later use.

[0045] Sterilization of empty seed tanks and fermentation tanks: Close all valves, open the drain valve at the bottom of the tank and the drain valve in the tank jacket, open the direct steam valve to introduce steam at 0.12~0.14MPa into the tank, start timing when the temperature inside the tank reaches 121℃, sterilization time is 40min, then close the drain valve at the bottom of the tank and the drain valve in the jacket, the temperature inside the tank will naturally drop to 37℃ for standby.

[0046] Formula: 3% lactose, 1% glucol, 0.5% dextran, 0.4% casein peptone, 0.3% potassium dihydrogen phosphate, 0.8% sodium acetate, 0.2% arginine, 0.1% Tween-80, 0.1% polyether defoamer, 93.6% tap water, natural pH.

[0047] Specific operation: First, put the tap water used in the formula into the sterilized seed tank. Turn on the mixer and add the raw materials required in the formula (the order of addition is not important). Then, stir continuously and introduce steam for heating. First, use steam in the tank jacket to heat to 95°C, then switch to direct steam heating. When the temperature in the tank reaches 121°C, maintain it for 20 minutes to achieve the sterilization effect. Then, turn off the steam and start cooling down by using tap water in the tank jacket. When the temperature inside the tank reaches 37°C, it is ready for use. This is called seed culture medium. Then, under aseptic conditions, inoculate the prepared primary bacterial culture solution into the seed tank. The inoculation amount is 5% of the culture medium in the tank. For every 100 kg of seed tank culture medium, 5 kg of primary bacterial solution is added. Then, cover the tank with the inoculation cap and start stirring and culturing. The culturing conditions are: temperature 37℃, pH value at rest, stirring speed 80 r / min, tank pressure 0.05 MPa, and culturing time 15 h to achieve the required conditions. If the tank pressure drops, sterile air can be introduced to maintain the tank pressure. If the tank pressure is too high, the vent valve on the top of the tank can be used to adjust it. This is called the seed tank bacterial culture.

[0048] IV. Fermentation Culture Formula: 4% lactose, 0.8% casein peptone, 1% dextran, 1.7% raffinose, 1.2% yeast extract, 0.3% sodium acetate, 0.2% Tween-80, 1% soybean protein powder, 0.9% dipotassium hydrogen phosphate, 0.1% polyether defoamer, and 88.8% tap water.

[0049] Specific operation: First, add the tap water used in the formula to the sterilized fermentation tank. Start the mixer and add the raw materials used in the formula to the fermentation tank (the order of addition is not important). Then, introduce steam for heating. First, use jacketed steam to heat to 95℃, then switch to direct steam heating. When the temperature in the tank reaches 121℃, start timing and maintain it for 30 minutes to achieve sterilization. Then, cool down the tank using tap water in the jacket. When the temperature in the tank drops to 37℃, start inoculating with the inoculum. Before inoculation, first increase the pressure in the seed tank to 0.1MPa, and maintain the pressure in the fermentation tank at 0.05MPa. Then, open the inoculation pipeline valve and transfer the cultured seeds from the seed tank to the fermentation tank through pressure difference. Then, close the inoculation channel valve.

[0050] The inoculum amount is 6 kg of inoculum cultured in the seed tank for every 100 kg of culture medium in the fermenter. The culture conditions are: culture temperature 37℃, stirring speed 140 r / min, tank pressure maintained at 0.05 MPa. During the fermentation process, the pH value is maintained at 6.5 through an automatic acid and alkali control system. If the tank pressure drops, sterile air can be introduced to maintain the tank pressure until the fermentation is completed. The fermentation lasts for 15 hours, and then the temperature is lowered to 20℃. This is the fermentation broth.

[0051] V. Inactivation and freeze-drying 1. Perform UHT inactivation treatment at 120℃ for 6 seconds.

[0052] 2. The fermentation broth is pumped into a storage tank for centrifugation. Operation: Start the centrifuge, adjust the speed to 10000 r / min, and centrifuge at a feed rate of 2 kg / min. After centrifugation, collect the metabolites of the inactivated wet sludge and freeze-dry them.

[0053] 3. Freeze-drying of mycelial sludge metabolites Preparation of bacterial suspension: Take 1000g of inactivated wet bacterial mud metabolites, add 4000mL of sterile physiological saline, 800g of lactose, 1000g of astragalus powder, 500g of jujube seed powder, and 30g of fucoidan.

[0054] Procedure: First, take 2500mL of physiological saline from the formula, add lactose and stir to dissolve, then add 1000g of inactivated wet bacterial sludge metabolites, then add the remaining physiological saline, astragalus powder and excipients from the formula and stir thoroughly to form a suspension. Freeze-dry the suspension until the moisture content is controlled at 4%, which becomes the freeze-dried bacterial powder of inactivated bacterial metabolites.

[0055] Tests showed that the freeze-dried bacterial powder contained 26% extracellular polysaccharides, 7% polypeptides, 22% amino acids, and 100 billion inactivated bacteria per gram.

[0056] Experimental Example 1 1. Metabolomics analysis The contents of astragaloside A, spinosin, and psoralen in the feces of lyophilized bacterial powder and mice (the mice were divided into a control group and an experimental group. The control group was fed normally and given 1.0 mL of physiological saline by gavage once a day; the experimental group was fed normally and given 1.0 mL of lyophilized bacterial powder solution (30 mg of lyophilized bacterial powder prepared in Example 1 dissolved in 1.0 mL of sterile water) by gavage once a day. After ten days of feeding, the contents of astragaloside A, spinosin, and psoralen were determined. The contents are shown in Table 1.

[0057] Determination of Astragaloside A content: High performance liquid chromatography Spinosin content detection method: liquid chromatography-tandem mass spectrometry Method for detecting psoralen content: High performance liquid chromatography (HPLC) Table 1. Content of astragaloside A, spinosin, and psoralen in the feces of mice fed with freeze-dried bacterial powder

[0058] 2. Effects on chronic nonbacterial prostatitis Fifty SD rats were randomly divided into a control group (n=10), a model group (n=10), and an experimental group (n=30). Rats in both the model and experimental groups were anesthetized with an intraperitoneal injection of 2.5% sodium pentobarbital (0.3 mL / 100 g), and under sterile conditions underwent castration. The skin was sutured, disinfected, and bandaged. The rats were then returned to their cages and allowed free access to food. On the second day, rats received a subcutaneous injection of estradiol benzoate (0.25 mg / kg) in their backs for 30 consecutive days. Rats in the control group received a subcutaneous injection of distilled water (0.125 mg / kg) for 30 consecutive days.

[0059] After successful model establishment, the experimental groups were randomly divided into three groups: experimental group 1 (n=10), experimental group 2 (n=10), and experimental group 3 (n=10). The control group and the model group were fed normally while being administered 1.0 mL of physiological saline by gavage. The experimental groups were fed normally while being administered 1.0 mL of lyophilized bacterial powder solution (20, 30, and 40 mg of the lyophilized bacterial powder prepared in Example 1 dissolved in 1.0 mL of sterile water) by gavage once daily for 30 days.

[0060] After 30 days of gavage administration, rats in each group were anesthetized by intraperitoneal injection of 2.5% sodium pentobarbital (0.3 mL / 100 g). Under aseptic conditions, the rats were dissected through a midline incision in the lower abdomen, reaching the peritoneal cavity. The prostate gland was removed, rinsed thoroughly with physiological saline, and weighed. 0.5 mL of physiological saline was added per gram of prostate tissue, and the mixture was homogenized using a glass homogenizer on an ice-water bath. After centrifugation at 3000 rpm for 20 min, the supernatant was collected. The levels of tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), and interleukin-10 (IL-10) in the prostate tissue homogenate were determined using enzyme-linked immunosorbent assay (ELISA). The test results are as follows: Figure 1 As shown.

[0061] according to Figure 1 It was found that the levels of TNF-α, IL-8, and IL-10 in the prostate tissue of rats in the model group were significantly higher than those in the control group. The levels of TNF-α, IL-8, and IL-10 in the prostate tissue of rats in each experimental group were significantly lower than those in the model group. This indicates that the freeze-dried bacterial powder can improve the inflammatory state of rat tissues with experimental chronic nonbacterial prostatitis and reduce the inflammatory effects of local inflammatory cytokines.

[0062] 3. Effects on benign prostatic hyperplasia Fifty specific pathogen-free (SPF) grade male Kunming mice were used. Ten mice were randomly selected as the control group, and the other four groups (n=10 each) were used to establish a mouse BPH model. These groups were designated as the model group, experimental group 1, experimental group 2, and experimental group 3. All mice in the model group were subcutaneously injected daily with 5 mg / kg of testosterone propionate (dissolved in soybean oil), while the control group received an equal volume of the solvent. This treatment continued for three weeks. In addition to normal feeding, the experimental groups were administered 1.0 mL of lyophilized bacterial powder solution (20, 30, and 40 mg of the lyophilized bacterial powder prepared in Example 1 dissolved in 1.0 mL of sterile water, respectively) via gavage. The model group and control group were administered the same volume of 0.5% sodium carboxymethyl cellulose solution (CMC solution) via gavage simultaneously with normal feeding. All treatments were administered once daily for three weeks.

[0063] ① Two hours after the last gavage (with fasting but free access to water for 12 hours), the mice were weighed, then euthanized by cervical dislocation, and the prostate was quickly removed. The wet weight of the prostate was measured, and the prostate index was calculated.

[0064] Prostate index = Prostate wet weight (mg) / Mouse body weight (g) ② Two hours after the last gavage (with fasting but free access to water for 12 hours), blood was collected from the orbital cavity of the mice. The blood was centrifuged at 3500 r / min for 10 min to separate the serum. The serum levels of testosterone (T), dihydrotestosterone (DHT), and estradiol (E2) were measured according to the kit instructions.

[0065] ③ The prostate was fixed in 10% formaldehyde solution, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE staining). The morphological changes of the prostate in each group of mice were compared.

[0066] Prostate index in mice, such as Figure 2 As shown. According to Figure 2 It can be seen that the prostate index in the model group was significantly higher than that in the control group, and all experimental groups could significantly reduce the prostate index in mice.

[0067] Mouse serum T level test results as follows Figure 3 As shown; DHT and E2 level test results are as follows Figure 4 As shown. From Figure 3 ,4 It can be seen that, compared with the control group, the serum T and DHT levels in the model group were significantly increased, and the serum E2 level was significantly decreased. Compared with the model group, all experimental groups significantly reduced the serum DHT and T levels and significantly increased the serum E2 level in mice.

[0068] Table 2 shows the effects on prostate tissue morphology in BPH model mice. As shown in Table 2, the model group exhibited significant pathological changes of benign prostatic hyperplasia, indicating successful model establishment. Compared with the model group, each experimental group significantly alleviated the pathological changes of benign prostatic hyperplasia in the model mice.

[0069] Table 2. Effects on prostate tissue morphology in BPH model mice (n=10)

[0070] Note: Grade 0: Normal glandular epithelium and stroma; Grade I: Hyperplasia of glandular epithelium, columnar in shape, occasionally with pseudopapillary formation or a small amount of fibrous connective tissue hyperplasia and smooth muscle hyperplasia in the stroma; Grade II: Hyperplasia of glandular epithelium, with a small number of papillae or pseudopapillaries and a small amount of fibrous connective tissue and smooth muscle hyperplasia in the stroma; Grade III: Hyperplasia of glandular epithelium, with more papillae or pseudopapillary protrusions, and significant hyperplasia of fibrous connective tissue and smooth muscle in the stroma, appearing as wide bands.

[0071] 4. Effects on sexual function and renal function in mice with kidney deficiency Fifty male ICR mice were randomly divided into 5 groups of 10 mice each: control group, model group, experimental group 1, experimental group 2, and experimental group 3. The control group was fed normally, while the model group and each experimental group were fed with 4 mg / kg / day. -1 Diluted estradiol benzoate was administered intraperitoneally once daily for 16 consecutive days. In the renal deficiency state, each experimental group was administered 1.0 mL of lyophilized bacterial powder solution (20, 30, and 40 mg of the lyophilized bacterial powder prepared in Example 1 dissolved in 1.0 mL of sterile water) by gavage once daily for 16 consecutive days. The control and model groups were administered the same volume of distilled water by gavage. Fifty ICR female mice were subcutaneously injected with estradiol benzoate solution at a dose of 20 µg / mouse 48 hours before the mating experiment to induce estrus. Male mice from all three groups were placed in cages for 3 minutes to acclimatize, one male at a time, followed by one estrus female mouse, to observe the mice's sexual function indicators.

[0072] ①During the experiment, the weight of the mice was measured every 4 days, and the condition of the mice on the 12th day after gavage was observed.

[0073] ② On day 11 after gavage, a mating experiment was conducted to detect the capture latency, mounting latency, ejaculation latency, number of captures, number of mountings, and number of ejaculations in mice.

[0074] Capture behavior: Male rats follow and chase female rats, and exhibit sniffing and licking behavior towards the female's genitals; Mounting behavior: The sexual behavior in which the male mouse mounts the back of the female mouse and moves its waist back and forth. Ejaculation behavior: After mounting behavior, the male rat will lick its penis.

[0075] The number of capture, mounting, and ejaculation behaviors by the male rat was defined as the number of times these behaviors occurred within 20 minutes of the female rat being placed in the female rat's enclosure.

[0076] The incubation period for capture, mounting, and ejaculation is the time from when the female mouse is introduced until the male mouse first exhibits capture, mounting, and ejaculation behaviors.

[0077] ③ Thirty minutes after the last gavage, load the base of the mouse's tail with an iron block equal to 10% of its body weight, and place the mouse in a swimming tank with a water depth of 35 cm and a water temperature of 25±1℃. Use a stopwatch to record the time from when the mouse starts swimming until it finally sinks to the bottom and dies; this is the swimming survival time.

[0078] ④ After the last gavage, blood was collected once from the fundus venous plexus of the mouse (the mouse was fasted for 12 hours before blood collection but allowed to drink water). The serum was collected by centrifugation, and the serum creatinine (Cr) and blood urea nitrogen (BUN) levels were measured.

[0079] ⑤ After the last gavage, blood was collected once from the fundus venous plexus of the mouse (the mouse was fasted for 12 hours before blood collection but allowed to drink water). The serum was collected by centrifugation, and the levels of T, luteinizing hormone (LH), follicle-stimulating hormone (FSH), E2, and gonadotropin-releasing hormone (GnRH) were measured. Seminal vesicle tissue was weighed, cut into small pieces, and added to pre-cooled 0.9% physiological saline at a volume ratio of 1:9. The mixture was homogenized, centrifuged, and the supernatant was collected to prepare a 10% seminal vesicle homogenate. The fructose and acid phosphatase (ACP) contents were measured.

[0080] ⑥ After the mice were euthanized, the penile tissue was weighed, cut into small pieces, and added to pre-cooled 0.9% physiological saline at a volume ratio of 1:9. The mixture was homogenized, centrifuged, and the supernatant was collected to prepare a 10% penile homogenate. The activities of nitric oxide synthase (NOS), phosphodiesterase type 5 (PDE-5), nitric oxide (NO), and cyclic guanosine monophosphate (cGMP) were then tested.

[0081] The body weight of mice in each group was statistically analyzed, and the results are shown in Table 3. Compared with the control group, the mice in the model group experienced a decrease in body weight due to kidney deficiency, with a significant decrease in body weight on day 12. In contrast, the experimental groups showed an increase in body weight compared to the model group, approaching the body weight of the control group. This indicates that the freeze-dried bacterial powder product alleviated the weight loss caused by kidney deficiency to some extent.

[0082] Compared with the control group, the model group mice exhibited symptoms of kidney deficiency, such as lethargy, hunching over, slow movement, sluggish response, and sweating. These same symptoms were present in all experimental groups before gavage administration, indicating the successful establishment of the kidney deficiency model. On day 12 after gavage administration, the kidney deficiency symptoms in all experimental groups were alleviated compared to the model group.

[0083] Table 3 Effects on mouse body weight (n=10)

[0084] Figure 5 This is a diagram showing the effect of the present invention on the capture latency of mice; Figure 6 This is a diagram showing the effect of the present invention on the mounting latency and ejaculation latency in mice; Figure 7 This is a graph showing the effect of the present invention on the number of mouse captures, mountings, and ejaculations. According to... Figure 5 , Figure 6 , Figure 7 It was found that, compared with the control group, the model group mice had longer capture latency, mounting latency, and ejaculation latency, and fewer capture, mounting, and ejaculation attempts. Compared with the model group mice, the experimental groups mice had shorter capture latency, mounting latency, and ejaculation latency, and although the number of capture, mounting, and ejaculation attempts increased, the differences in mounting and ejaculation attempts were not significant. This indicates that freeze-dried bacterial powder products can reduce the damage caused by kidney deficiency and restore sexual function in mice.

[0085] Table 4 shows the comparison results of survival time in swimming experiments of mice in different groups. According to Table 4, compared with the control group, the survival time of mice in the model group during weight-bearing swimming was significantly shorter. After gavage, the survival time of mice in each experimental group during weight-bearing swimming was significantly longer, indicating that probiotics have a significant anti-fatigue effect.

[0086] Table 4 Comparison of survival time of mice in different groups during swimming experiment (n=10)

[0087] Figure 8 This is a graph showing the effect of the present invention on serum Cr concentration; Figure 9 This is a graph showing the effect of this invention on serum BUN concentration. Serum Cr and BUN concentrations are two important indicators for evaluating whether kidney function is normal. Figure 8 , 9 As shown, the concentrations of Cr and BUN in the serum of mice in the model group were significantly higher than those in the control group. After gavage, the concentrations of Cr and BUN in the serum of mice decreased. The freeze-dried bacterial powder product can reduce the concentrations of Cr and BUN in mouse serum, thus reducing the kidney damage they cause.

[0088] Figure 10This is a diagram showing the effect of the present invention on LH, FSH, and E2 levels; Figure 11 This is a diagram showing the effect of the present invention on GnRH levels; Figure 12 This is a graph showing the effect of this invention on T levels. According to... Figure 10 It was found that the levels of LH, FSH, and E2 in the model group mice were significantly higher than those in the control group mice. After gavage, the levels of LH, FSH, and E2 in the mice significantly decreased. Figure 11 , Figure 12 It was found that the GnRH and T levels in the model group mice were significantly lower than those in the control group mice. The GnRH and T levels in each experimental group mice were significantly higher than those in the model group mice, and recovered to levels close to those in the control group mice. This indicates that kidney deficiency leads to hormonal imbalances in mice, and that the freeze-dried bacterial powder product can restore this homeostasis.

[0089] Figure 13 This diagram illustrates the effects of this invention on ACP in the prostate and fructose in the seminal vesicles of mice. Changes in ACP concentration in seminal plasma are important indicators for understanding prostate function and diagnosing prostate diseases. Changes in fructose concentration can indicate the presence of seminal vesicle diseases. Based on... Figure 13 The results showed that the concentration of ACP in the seminal plasma of the model group mice was significantly higher than that of the control group mice, while the concentration of fructose was the opposite. Compared with the model group mice, the concentration of ACP in the seminal plasma of each experimental group mice was significantly lower, while the concentration of fructose was higher, but the difference was not significant. This indicates that the freeze-dried bacterial powder product can treat prostate diseases and seminal vesiculitis by reducing the concentration of ACP in the prostate and increasing the concentration of fructose in the seminal vesicles.

[0090] Figure 14 This is a graph showing the effect of the present invention on NO and cGMP in the mouse penis; Figure 15 This is a diagram showing the effect of the present invention on NOS in the mouse penis; Figure 16 This is a graph showing the effect of this invention on PDE-5 in the mouse penis. The concentrations of NO and cGMP, and the activities of NOS and PDE-5 in the penile homogenate are key factors in the physiological process of male penile erection. According to... Figure 14 , Figure 15 , Figure 16 It was found that the concentrations of NO, cGMP, and NOS activity in the penile homogenate of the model group mice were significantly lower than those in the control group, while the activity of PDE-5 was the opposite. After gavage, compared with the model group mice, the concentrations of NO, cGMP, and NOS activity in the penile homogenate of each experimental group mice were significantly increased, while the activity of PDE-5 was the opposite. This indicates that the freeze-dried bacterial powder product can inhibit the decrease in NO, cGMP concentration and NOS activity, increase PDE-5 activity, and restore the sexual function of mice.

[0091] 5. The recruitment targets were males with an average age of 35. As of August 2025, a total of 120 people were recruited to participate in the experiment. All of the above-mentioned individuals signed written informed consent forms. If a participant was unable to sign, an agent (usually a family member) would sign. The basic information of the participants is shown in Table 5.

[0092] Table 5. Basic Information on Participants' Participation in the Experiment

[0093] Prostatitis: Results were collected from 30 participants after 8 days of use. After 8 days, 8 participants experienced improvement in symptoms such as poor mental state, insomnia, and burning or stinging sensations during urination. After 16 days, 15 participants experienced significant improvement in these symptoms. After 24 days, 22 participants experienced extremely significant improvement, with reduced urination frequency and increased sexual interest. After 30 days, 27 participants experienced complete improvement in these symptoms, with normal urination frequency, normal sexual activity, and restored libido. Three participants did not experience improvement after taking Lactococcus lactis.

[0094] Benign Prostatic Hyperplasia (BPH): Results were collected from 30 participants after 8 days of use. After 8 days, 7 participants experienced improvement in symptoms such as dribbling urination, slow urination, incomplete emptying of the bladder, weak urine stream, increased nighttime urination, and poor sleep quality. After 16 days, 11 participants experienced significant improvement in these symptoms. After 24 days, 20 participants experienced extremely significant improvement. After 30 days, 26 participants experienced complete improvement in these symptoms. Four participants did not experience any improvement after taking Lactococcus lactis.

[0095] Kidney Deficiency: Results were collected from 30 participants after 15 days of use. After 8 days, 5 participants experienced improvement in symptoms such as fatigue, lethargy, low energy, easy fatigue; aversion to cold, cold limbs, heaviness in the body; lower back and knee pain, cold back pain, muscle weakness; and decreased sexual function. After 30 days, 11 participants experienced significant improvement in these symptoms. After 45 days, 20 participants experienced extremely significant improvement in symptoms such as fatigue, lethargy, low energy, easy fatigue; aversion to cold, cold limbs, heaviness in the body; lower back and knee pain, cold back pain, muscle weakness; and decreased sexual function, impotence, and premature ejaculation. After 60 days of use, 24 people experienced complete improvement in symptoms such as fatigue, lethargy, low energy, and easy exhaustion; cold intolerance, cold limbs, and heaviness in the body; lower back and knee pain, cold pain in the lower back, and weakness in the muscles and bones; and decreased sexual function, impotence, and premature ejaculation. Six people did not experience any improvement after taking Lactococcus lactis.

[0096] Experimental Example 2 1. Experiment on regulating the function of mouse gut microbiota Mice were divided into a control group and an experimental group, with 10 mice in each group. The control and model groups were given 1.0 mL of physiological saline by gavage once daily, in addition to normal feeding. The experimental group was given 1.0 mL of lyophilized bacterial powder solution (30 mg of lyophilized bacterial powder prepared in Example 1 dissolved in 1.0 mL of sterile water) by gavage once daily, in addition to normal feeding. After 30 days of gavage, mouse feces were aseptically collected, weighed, and diluted 10-fold with sterile diluent to prepare a homogeneous suspension. This suspension was then serially diluted 10-fold, and appropriate dilutions were selected and inoculated onto various selective culture media. The number of bacteria per gram of wet feces was calculated, and the logarithm was used for statistical analysis. Twenty-four hours after the last administration of the lyophilized bacterial powder, rectal feces were collected to detect the intestinal flora. The results are shown in Table 6.

[0097] Table 6. Suitable culture conditions and detection of gut microbiota (x - ±SD)

[0098] As can be seen from Table 6, the freeze-dried bacterial powder product can increase the number of beneficial bacteria (Bifidobacterium and Lactobacillus), while the number of harmful bacteria (Enterobacterium and Enterococcus) does not increase but instead shows a slight decreasing trend, indicating that the freeze-dried bacterial powder product prepared in Example 1 of this invention has the function of regulating the intestinal flora of mice.

[0099] 2. Mouse gastrointestinal motility test Mice were divided into a control group, a model group, and an experimental group, with 10 mice in each group. The control and model groups were given 1.0 mL of sterile water by gavage while being fed normally, once daily. The experimental group was given 1.0 mL of lyophilized bacterial powder solution (30 mg of lyophilized bacterial powder prepared in Example 1 dissolved in 1.0 mL of sterile water) by gavage while being fed normally, once daily. After 15 days of gavage, all mice were fasted for 16 hours. Then, the model and experimental groups were given compound diphenoxylate by gavage, while the control group was given distilled water by gavage. 30 minutes later, all mice were given gum arabic ink by gavage. 30 minutes later, the mice were euthanized by cervical vertebrae amputation. The abdomen was quickly incised along the midline to separate the mesentery. The intestine was cut from the pylorus to the ileocecal junction, and the small intestine was gently pulled into a straight line. The length of the intestine was measured as the "total length of the small intestine," and the distance from the pylorus to the ink tip was measured as the "ink propulsion length." The ink propulsion rate was calculated using the following formula. The small intestine ink propulsion rate is shown in Table 7.

[0100] ; The units for both ink propulsion length and small intestine propulsion length are centimeters.

[0101] Table 7. Intestinal ink propulsion rate (x) - ±SD)

[0102] As shown in Table 7, the mouse intestinal peristalsis inhibition model was successfully established by administering compound diphenoxylate. Compared with the model group, the small intestinal ink propulsion rate increased in the experimental group, indicating that the freeze-dried bacterial powder product prepared in Example 1 of this invention can promote small intestinal peristalsis and digestion.

[0103] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A type of *Pediococcus lactis* NH3 (for treating male prostate issues and improving sexual function) Pediococcus acidilactici ), characterized in that, This strain was deposited at the Guangdong Provincial Center for Microbial Culture Collection on April 18, 2025, with accession number GDMCC No: 66163.

2. The *Pediococcus lactis* NH3 as described in claim 1 Pediococcus acidilactici The cultivation method of ) is characterized by, The cultivation methods include: scale-up culture, primary seed culture, seed tank culture, and fermentation culture.

3. The cultivation method as described in claim 2, characterized in that, The culture medium for expansion culture contains, per 1000 mL of water: 15-25 g lactose, 3.25-3.75 g raffinose, 4-6 g inulin, 2-5 g glucosyl alcohol, 0.5-2 g sodium acetate, 0.5-1 mL Tween-80, and 0.6-1 g dipotassium hydrogen phosphate; the pH of the culture medium for expansion culture is adjusted to 6.0-6.5; the culture conditions for expansion culture are static incubation at 36-38℃ for 10-20 h. Alternatively, the culture medium for primary seed culture contains the following per 1000 mL of water: lactose 2-5 g, glucose 1-3 g, casein peptone 4-6 g, inulin 1-3 g, beef extract 2-4 g, arginine 1-3 g, sodium acetate 0.5-2 g, threonine 0.5-2 g, and Tween-80 0.5-1 mL; the pH of the culture medium for primary seed culture is natural; the culture conditions for primary seed culture are 36-38℃ for 20-30 h; the inoculum size for primary seed culture is 0.4-0.6%, by volume. Alternatively, the culture medium for seed tank culture may contain, by weight percentage: lactose 2-4%, glucol 0.5-1.5%, dextran 0.5-1%, casein peptone 0.3-0.6%, potassium dihydrogen phosphate 0.2-0.5%, sodium acetate 0.6-1.0%, arginine 0.1-0.3%, Tween-80 0.1-0.2%, polyether defoamer 0.05-0.15%, and tap water 92-94%; the pH of the culture medium for seed tank culture should be natural; the culture conditions for seed tank culture are: tank pressure 0.04-0.06 MPa, culture at 36-38℃ for 10-20 h; the inoculum size for seed tank culture is 4-6%, by volume. Alternatively, the fermentation culture medium may contain, by weight percentage: lactose 3-5%, casein peptone 0.6-1.0%, dextran 0.5-1.5%, raffinose 1.5-2.0%, yeast extract 1.0-1.5%, sodium acetate 0.2-0.5%, Tween-80 0.1-0.3%, soybean protein powder 0.5-1.5%, dipotassium hydrogen phosphate 0.8-1.0%, polyether defoamer 0.05-0.15%, and tap water 87-89%; the pH of the fermentation culture medium is 6.0-7.0; the fermentation conditions are: tank pressure 0.04-0.06 MPa, temperature 36-38℃, and time 10-20 h.

4. The cultivation method as described in claim 3, characterized in that, The culture medium for expansion culture contains, per 1000 mL of water: 20.0 g lactose, 3.50 g raffinose, 5.0 g inulin, 3.0 g g glucosyl alcohol, 1.0 g sodium acetate, 0.5 mL Tween-80, and 0.8 g dipotassium hydrogen phosphate; the pH of the culture medium for expansion culture is adjusted to 6.0; the culture conditions for expansion culture are static incubation at 37℃ for 16 h. Alternatively, the primary seed culture medium contains, per 1000 mL of water: 3 g lactose, 2 g glucose alcohol, 5 g casein peptone, 2 g inulin, 3 g beef extract powder, 2 g arginine, 1 g sodium acetate, 1 g threonine, and 1 mL Tween-80; the pH of the primary seed culture medium is natural; the primary seed culture conditions are 37°C for 25 h; the inoculum size for the primary seed culture is 0.5% by volume. Alternatively, the culture medium for seed tank culture comprises, by weight percentage: 3% lactose, 1% glucan, 0.5% dextran, 0.4% casein peptone, 0.3% potassium dihydrogen phosphate, 0.8% sodium acetate, 0.2% arginine, 0.1% Tween-80, 0.1% polyether defoamer, and 93.6% tap water; the pH of the culture medium for seed tank culture is natural; the culture conditions for seed tank culture are: tank pressure 0.05 MPa, culture at 37°C for 15 h; the inoculum size for seed tank culture is 5% by volume. Alternatively, the fermentation culture medium may, by weight percentage, comprise: 4% lactose, 0.8% casein peptone, 1% dextran, 1.7% raffinose, 1.2% yeast extract, 0.3% sodium acetate, 0.2% Tween-80, 1% soybean protein powder, 0.9% dipotassium hydrogen phosphate, 0.1% polyether defoamer, and 88.8% tap water; the pH of the fermentation culture medium is 6.5; the fermentation conditions are: tank pressure 0.05 MPa, temperature 37°C, and time 15 h.

5. A microbial inoculant, characterized in that, It includes an active ingredient, which includes the *Pediococcus lactis* NH3 or its fermentation broth as described in claim 1.

6. The use of the *Pediococcus lactis* NH3 as described in claim 1 or the microbial agent as described in claim 5 in the preparation of health foods that help regulate intestinal flora or aid digestion.

7. The use of the *Pediococcus lactis* NH3 as described in claim 1 or the microbial agent as described in claim 5 in the preparation of a medicament for treating nonbacterial prostatitis, benign prostatic hyperplasia, and kidney deficiency.

8. The application as described in claim 7, characterized in that, The drug exists in the form of lyophilized bacterial powder.

9. A method for preparing freeze-dried bacterial powder for treating nonbacterial prostatitis, benign prostatic hyperplasia, and kidney deficiency, characterized in that, include: The fermentation broth obtained after fermentation culture according to claim 2 is inactivated and centrifuged to obtain the following: The inactivated wet bacterial sludge metabolites were freeze-dried, then mixed with excipients to prepare a suspension, and the suspension was freeze-dried to obtain the final product.

10. The method as described in claim 9, characterized in that, Inactivation refers to UHT inactivation treatment of fermentation broth at 100~140℃ for 5~8s. Alternatively, centrifuge at a speed of 8000~12000 r / min; centrifuge at a feed rate of 1~3 kg / min; Alternatively, the moisture content of the freeze-dried bacterial powder is ≤4% by mass. Alternatively, the excipients may include lactose, astragalus powder, jujube seed powder, and fucoidan; Alternatively, the mass ratio of inactivated wet bacterial sludge metabolites to auxiliary materials may be 1:2~2.5.

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