Lactobacillus crispatus and application thereof in prevention and treatment of female reproductive system diseases

Abstract

The invention discloses lactobacillus crispatus, a microbial inoculum and a preparation method and application of the lactobacillus crispatus and the microbial inoculum. The lactobacillus crispatus disclosed by the invention is lactobacillus crispatus OH126 of which the preservation number is GDMCC (China General Microbiological Culture Collection Center) No.66536. The lactobacillus crispatus newly researched and developed in the invention can effectively inhibit growth of common vaginal pathogenic bacteria, is adaptive to a vaginal micro-ecological environment, has good vaginal mucosa colonization ability, and can also assist in improving related symptoms of hysteromyoma. Meanwhile, vaginal freeze-dried tablets, vaginal gel and other microbial agents can be prepared, and the activity and storage stability of thalli are ensured by optimizing the preparation process. The strain and the microbial inoculum can be used for adjusting female vagina micro-ecological balance, treating or preventing bacterial vaginitis and assisting in improving related symptoms of hysteromyoma, and a new scheme and a new way are provided for prevention and treatment of female reproductive system diseases such as bacterial vaginitis and hysteromyoma.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of lactobacillus crispatus, in particular to a lactobacillus crispatus, a bacterial agent containing the strain and a preparation method thereof, and application of the lactobacillus crispatus and the bacterial agent in preparation of products for preventing and treating female reproductive system diseases (especially bacterial vaginosis and uterine fibroids). BACKGROUND

[0002] Female reproductive system diseases are common diseases that threaten the health of women all over the world, among which bacterial vaginosis (BV) and uterine fibroids are two diseases with high incidence and far-reaching influence. According to epidemiological data, the incidence of bacterial vaginosis in women of childbearing age is about 11%-33% according to different races. Patients have vaginal microecological imbalance, which is manifested as increased vaginal discharge, odor and pruritus of vulva. If it recurs for a long time, it can increase the risk of pelvic inflammation and urinary tract infection, and infection during pregnancy can also lead to adverse pregnancy outcomes such as premature delivery and premature rupture of membranes; uterine fibroids are the most common pelvic tumors in women of childbearing age, affecting one in four adult women. Some patients are accompanied by menstrual abnormalities, abdominal pain, anemia and other symptoms, and severe cases require surgical intervention, which not only affects the quality of life, but also may interfere with the reproductive function.

[0003] Under healthy conditions, lactobacillus crispatus is the absolute dominant bacteria in the vaginal microecosystem of women, can produce immunomodulatory and antibacterial compounds, can inhibit inflammation and help maintain the mucosal barrier, thereby preventing pathogen colonization. It can maintain an acidic environment with pH<4.5 in the vagina by metabolizing lactic acid, inhibit the growth of pathogenic bacteria such as gardnerella and prevotella; at the same time, it can secrete bacteriocins, hydrogen peroxide and other antibacterial substances to directly destroy the cell membrane structure and enzyme activity of pathogenic bacteria, and form a biofilm barrier by competitively adhering to vaginal epithelial cells to prevent pathogenic bacteria from colonizing. In addition, the rapid growth characteristics of high-quality lactobacillus crispatus can quickly occupy the ecological niche of the vagina, further consolidating the microecological balance. However, there are significant limitations in current prevention and treatment methods for female reproductive system diseases: in the treatment of bacterial vaginosis, antibiotics such as metronidazole and clindamycin are commonly used in clinical practice, which can kill pathogenic bacteria in the short term, but will indiscriminately eliminate beneficial lactobacillus crispatus in the vagina, exacerbate microecological imbalance, lead to an increased recurrence rate of the disease, and long-term use can easily lead to drug resistance of pathogenic bacteria; for uterine fibroids, the existing treatment mainly relies on hormone drugs to inhibit the growth of fibroids or surgical resection, and hormone drugs have side effects such as weight gain and mood swings, and surgery causes trauma to the reproductive system, and there is a lack of effective means to improve the condition by adjusting the local microenvironment.

[0004] Existing probiotic preparations for the female reproductive system also face technological bottlenecks: most products use *Lactobacillus curvatureii* strains isolated from the intestines or milk, rather than from the vagina of healthy women. These strains have poor adaptability to the acidic environment of the vagina, weak acid-producing ability, narrow antibacterial spectrum, and slow growth rate, making it difficult to quickly rebuild the dominant vaginal flora. Although some strains possess certain activity, the formulation process lacks targeted live bacteria protection technology. The survival rate of bacteria is low during freeze-drying, and the activity decays rapidly during storage, resulting in insufficient effective live bacteria after reaching the vagina, failing to fully exert the probiotic effect. More importantly, current probiotic preparations only focus on infectious diseases such as bacterial vaginosis, and there are no *Lactobacillus curvatureii* strains and products that can help improve symptoms related to uterine fibroids, thus failing to meet the needs of comprehensive prevention and treatment of female reproductive system diseases.

[0005] Therefore, developing a new strain of *Lactobacillus curvatureensis* derived from the vaginal microenvironment of healthy women, possessing high acid-producing capacity, strong antibacterial activity, and rapid growth characteristics, and capable of simultaneously assisting in the prevention and treatment of bacterial vaginosis and uterine fibroids, along with the development of a highly stable and well-preserved bacterial agent, is key to solving the current challenges in the prevention and treatment of female reproductive system diseases. This has significant clinical value and application prospects for improving disease intervention effects, reducing drug side effects, and maintaining women's reproductive health. Summary of the Invention

[0006] The purpose of this application is to provide a novel *Lactobacillus curvaturei* strain, a bacterial agent prepared from the *Lactobacillus curvaturei* strain, a method for preparing the bacterial agent, and the application of the *Lactobacillus curvaturei* strain and the bacterial agent in the prevention and treatment of female reproductive system diseases.

[0007] The following technical solution is adopted in this application: One aspect of this application discloses a strain of Lactobacillus curvature, which is Lactobacillus curvature OH126 with accession number GDMCC No. 66536.

[0008] It should be noted that the novel *Lactobacillus curvularis* strain mentioned in this application exhibits significant inhibitory effects on the growth of core pathogens of bacterial vaginosis, such as *Gardnerella vaginalis*, *Prevotella*, and *Morchella molybditis*, as well as common opportunistic pathogens in the vagina, such as *Candida albicans*. Experimental results show that its 30% culture supernatant exhibits an inhibition rate of 87%-92% against these pathogens, and the antibacterial effect increases with the increase in the proportion of supernatant, significantly higher than that of ordinary vaginal *Lactobacillus curvularis*. Simultaneously, it has a strong adhesion effect on vaginal epithelial cells, with an adhesion rate of 19.5%-22.0% on human vaginal epithelial cells VK2 / E6E7. Furthermore, it possesses high acid-producing capacity; after anaerobic culture at 37°C for 36 hours in MRS liquid medium, lactic acid production reaches 12.5-14 g / L, rapidly reducing the pH of the culture medium simulating the vaginal environment to 3.6-3.8. Furthermore, its cell-free fermentation supernatant can inhibit the proliferation activity of human uterine fibroid cells (HUMSCs) with an inhibition rate of 10%-12%, and can downregulate the expression levels of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) in fibroid tissue, while the fermentation supernatant of ordinary *Lactobacillus curvularia* has a weaker inhibitory effect on the proliferation of uterine fibroid cells. The *Lactobacillus curvularia* OH126 of this application can effectively treat or prevent bacterial vaginosis and help improve symptoms related to uterine fibroids (such as menstrual abnormalities and lower abdominal distension), providing a new direction for the prevention and treatment of female reproductive system diseases.

[0009] Another aspect of this application discloses a microbial agent containing the Lactobacillus curvature OH126 of this application.

[0010] Preferably, the bacterial agent of this application is a vaginal lyophilized tablet or vaginal gel.

[0011] It should be noted that the key to the bacterial agent of this application lies in containing *Lactobacillus curvaturei* OH126, as described in this application. Other components can be added according to requirements or dosage form. It is understood that *Lactobacillus curvaturei* OH126, as a probiotic adapted to the female vaginal microenvironment, can be used directly as the active ingredient of the bacterial agent of this application; alternatively, the fermentation products of *Lactobacillus curvaturei* OH126 can be used as the active ingredient of the bacterial agent of this application. For example, it can be the culture medium of *Lactobacillus curvaturei* OH126, or its derivative fermentation products, such as vaginal microecological regulation preparations made using the excellent acid-producing ability of *Lactobacillus curvaturei* OH126.

[0012] Another aspect of this application discloses a method for preparing the bacterial agent of this application, namely, a method for preparing lyophilized tablets for vaginal use, comprising the following steps: (1) Cell collection: Lactobacillus curvature OH126 was inoculated into MRS liquid medium (formula: peptone 10.0 g / L, beef extract 10.0 g / L, yeast extract 5.0 g / L, glucose 20.0 g / L, sodium acetate 5.0 g / L, diammonium citrate 2.0 g / L, Tween-80 1.0 g / L, dipotassium hydrogen phosphate 0.4 g / L, magnesium sulfate 0.58 g / L, manganese sulfate 0.29 g / L) and anaerobic cultured at 37℃ for 12-14 h until the late logarithmic growth phase (OD). 600 A value of 1.15-1.20 corresponds to a viable bacterial count of approximately 3.5 × 10⁻⁶. 9 -3.6×10 9 (CFU / mL), then centrifuged at 10000-12000 rpm for 15-20 min at 4℃, discarded the supernatant, and collected the live bacteria (bacterial sludge). The entire process was carried out under aseptic conditions. (2) Cell encapsulation: Add freeze-drying protectant to the bacterial sludge and mix well; (3) Freeze-drying: The bacterial cells with added freeze-drying protectant were dispensed into freeze-drying molds (0.5g per mold) and placed in a freeze dryer for freeze-drying. First, the cells were pre-frozen at -45℃ for 3-4 hours, then heated to -25℃ at a rate of 5℃ / h and held for 2 hours, then heated to -10℃ at a rate of 3℃ / h and held for 3 hours, and finally heated to 25℃ at a rate of 2℃ / h and held for 4 hours. The total freeze-drying time was 18-20 hours. The vacuum degree was maintained at 10-15 Pa during the freeze-drying process to obtain freeze-dried bacterial blocks. (4) Pulverization: The freeze-dried product is pulverized and passed through an 80-mesh sieve to obtain freeze-dried powder; (5) Mix the lyophilized powder with pharmaceutically acceptable excipients and compress it into tablets using a tablet press at a pressure of 20-25 kN to obtain a bacterial agent in the form of lyophilized tablets for vaginal use.

[0013] Preferably, the mass ratio of the freeze-drying protectant to the Lactobacillus curvature sludge is 2:1.

[0014] Preferably, the freeze-drying protectant includes at least one of skim milk powder, trehalose, and sucrose.

[0015] Preferably, the freeze-drying protectant contains 12 wt% skim milk powder, 8 wt% trehalose, and 5 wt% sucrose.

[0016] Preferably, pharmaceutically acceptable excipients include at least one of fillers, disintegrants, and lubricants.

[0017] Preferably, the vaginal lyophilized tablets of this application comprise 30-35 parts of lyophilized powder, 25-30 parts of filler, 5-8 parts of disintegrant, and 1-2 parts of lubricant.

[0018] Preferably, the filler is at least one of microcrystalline cellulose and lactose.

[0019] Preferably, the disintegrant is at least one of crospovidone and low-substituted hydroxypropyl methylcellulose.

[0020] Preferably, the lubricant is at least one of magnesium stearate and silicon dioxide.

[0021] Preferably, each of the lyophilized vaginal tablets contains ≥1×10⁻⁶ live Lactobacillus curvature. 9 CFU.

[0022] Another aspect of this application discloses another method for preparing the bacterial agent of this application, namely, a method for preparing vaginal gel, comprising the following steps: (1) Collection of bacterial cells: Collect live Lactobacillus curvature OH126 cells (bacterial sludge) by referring to the “Collection of bacterial cells” step in the preparation method of vaginal lyophilized tablets. (2) Preparation of bacterial suspension: Add sterile physiological saline to the bacterial sludge and adjust the concentration of the bacterial suspension to 1×10¹. 0 -2×10¹ 0 CFU / mL was used to obtain bacterial culture; (3) Gel matrix preparation: Add purified water to the gel matrix material and let it swell at room temperature for 24 hours. Stir 3-4 times during the period until a uniform and transparent gel matrix is ​​formed. Add humectant and preservative to the gel matrix, stir evenly, and then adjust the pH to 3.8-4.2 with pH adjuster. (4) Mixing: Under sterile conditions, slowly add the bacterial solution to the gel matrix while stirring at a rate of 50-60 rpm to ensure that the bacterial solution is evenly dispersed. Finally, add purified water to a total mass of 100g to form a vaginal gel. (5) Dispensing and sterilization: Dispense the prepared vaginal gel into sterile pharmaceutical gel tubes (5g per tube), and sterilize them by irradiation with γ-rays at a dose of 2-3kGy to obtain a bacterial agent in the form of vaginal gel.

[0023] It should be noted that the bacterial collection step in the preparation method of vaginal gel is the same as that in the preparation method of vaginal lyophilized tablets. Therefore, the bacterial collection step can be directly referred to the preparation method of vaginal lyophilized tablets in this application, and will not be repeated here.

[0024] It should be noted that, as needed, the bacterial sludge or bacterial solution can be encapsulated in enteric microcapsules to further enhance the stability of the bacteria in the vagina.

[0025] Preferably, the gel matrix material includes at least one of carbomer 940 and hydroxypropyl methylcellulose.

[0026] Preferably, the amount of gel matrix material added is 1.5-2.0g / 100g gel.

[0027] Preferably, the moisturizer includes at least one of glycerin and propylene glycol.

[0028] Preferably, the amount of moisturizer added is 5-8g / 100g gel.

[0029] Preferably, the preservative is phenoxyethanol.

[0030] Preferably, the amount of preservative added is 0.3-0.5g / 100g gel.

[0031] Preferably, the pH adjuster is at least one of citric acid and sodium citrate.

[0032] Preferably, each vaginal gel (5g / tube) contains ≥1×10⁻⁶ live Lactobacillus curvature. 9 CFU.

[0033] Another aspect of this application discloses the use of Lactobacillus curvature OH126 or the bacterial agent of this application in the preparation of products for the treatment or prevention of bacterial vaginosis; or in the preparation of products for the assistance in improving symptoms related to uterine fibroids; or in the preparation of products for regulating the vaginal microecological balance in women.

[0034] Another aspect of this application discloses a product for treating or preventing bacterial vaginosis, the product containing Lactobacillus curvularia OH126 of this application or the bacterial agent of this application.

[0035] Another aspect of this application discloses a product that helps improve symptoms related to uterine fibroids, the product containing Lactobacillus curvature OH126 or the bacterial agent of this application.

[0036] Another aspect of this application discloses a product for regulating the vaginal microecological balance in women, which contains Lactobacillus curvature OH126 or the bacterial agent of this application.

[0037] It should be noted that the *Lactobacillus curvature* OH126 of this application also possesses some common characteristics of existing *Lactobacillus curvature* species. Therefore, referring to existing *Lactobacillus curvature* species, the *Lactobacillus curvature* OH126 of this application can also be used to prepare health care products or medical devices for the female reproductive system. Furthermore, health care products or medical devices containing the *Lactobacillus curvature* OH126 of this application can inhibit the growth of vaginal pathogens such as *Gardnerella vaginalis*, *Prevotella vaginalis*, *Morchella molybditis*, and *Candida albicans*, and have effects such as treating or preventing bacterial vaginosis, assisting in improving symptoms related to uterine fibroids, and regulating the vaginal microecological balance.

[0038] The beneficial effects of this application are as follows: The newly developed *Lactobacillus curvatureii* OH126 in this application effectively inhibits the growth of pathogens associated with bacterial vaginosis and common opportunistic pathogens in the vagina, with significantly better antibacterial effects than ordinary *Lactobacillus curvatureii*. It also exhibits high acid production, rapid growth, and high vaginal epithelial cell adhesion, efficiently regulating the vaginal microecological balance, preventing and treating bacterial vaginosis, and addressing the problems of drug resistance and microecological disruption caused by existing antibiotic treatments. Furthermore, its fermentation supernatant can inhibit the proliferation of uterine fibroid cells, helping to improve symptoms related to uterine fibroids, filling a gap in the field of probiotics for the adjuvant improvement of uterine fibroids, and providing a new solution for the prevention and treatment of female reproductive system diseases such as bacterial vaginosis and uterine fibroids. The vaginal lyophilized tablets and gels prepared based on this strain, through optimized preparation processes, maximize the preservation of bacterial activity. Even after 12 months of storage at 4°C, the viable bacterial count remains above 80% of the initial value, and the dosage form is suitable for local vaginal application, ensuring effective colonization and efficacy of the strain in the vagina, overcoming the shortcomings of existing probiotic preparations such as poor adaptability, insufficient stability, and limited functionality. Attached Figure Description

[0039] Figure 1 This is a colony morphology diagram of Lactobacillus curvature OH126 strain after anaerobic culture on an MRS solid plate in Example 1 of this application; Figure 2 This is a growth curve of Lactobacillus curvature OH126 strain in Example 1 of this application (the horizontal axis represents culture time, and the vertical axis represents OD). 600 Value and viable count); Figure 3 This image shows the antibacterial effect of the culture supernatant of Lactobacillus curvaturei OH126 in Example 2 of this application against Gardnerella vaginalis, Prevotella, Molybditis lappa and Candida albicans. Figure 4 The effect of Lactobacillus curvature OH126 culture supernatant on the survival rate of RAW264.7 macrophages in Example 2 of this application; Figure 5 This is a survival rate curve of Lactobacillus curvature OH126 in Example 3 of this application under different pH values ​​(3.0-5.0) and different bile salt concentrations (0.1%-0.5%). Figure 6 This is a graph showing the effect of Lactobacillus curvaturei OH126 vaginal gel on the vaginal mucosal inflammation score in rats, as described in Example 7 of this application.

[0040] The Lactobacillus crispatus strain described in this application was deposited on June 16, 2025, at the Guangdong Provincial Center for Microbial Culture Collection, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with accession number GDMCC No. 66536. Detailed Implementation

[0041] Lactobacillus curvature is a core dominant bacterium in the vaginal microecology of healthy women, playing a crucial role in maintaining the acidic environment of the vagina and inhibiting the colonization of pathogenic bacteria. This application study discovered a novel Lactobacillus curvature strain, Lactobacillus curvature OH126, adapted to the needs of female reproductive health. This strain was isolated from vaginal secretions of healthy women of reproductive age (subjects had no history of reproductive system infection or organic disease, and had not used antibiotics, probiotics, or hormonal drugs within the past 3 months). It effectively inhibits common vaginal pathogens and possesses good vaginal mucosal colonization ability and tolerance to vaginal microenvironment stress, making it suitable for the development of probiotic products related to female reproductive health.

[0042] The *Lactobacillus curvature* OH126 of this application has the following properties: (1) The Lactobacillus curvatureis OH126 of this application has a significant inhibitory effect on common vaginal pathogens, including Gardnerella vaginalis ATCC 14018, Prevotella bivia ATCC 29303, Mobiluncus mulieris ATCC 35243 and Candida albicans ATCC10231. The antibacterial effect gradually increases with the increase of the proportion of its culture supernatant (10%-30%). At the same concentration (30%), the inhibitory effect is better than that of the control strain Lactobacillus acidophilus GLA-14. (2) The Lactobacillus curlis OH126 of this application has a moderate to superior hydrogen peroxide production capacity. After being coated on MRS plates containing TMB and horseradish peroxidase and exposed to air, the color change time is shorter than that of the control group Lactobacillus acidophilus GLA-14. (3) The Lactobacillus curvature OH126 of this application has a strong ability to produce lactic acid; (4) The *Lactobacillus curvature* OH126 of this application has good resistance to vaginal microenvironment stress. It can still maintain a high survival rate after being incubated for 2 hours in simulated healthy and abnormal vaginal acidic environments (pH 3.0-5.0). After being incubated for 4 hours in common vaginal secretion bile salt concentrations (0.1%-0.5%), its survival rate is significantly higher than that of *Lactobacillus acidophilus* GLA-14, which is adapted to the local physiological environment of the vagina. (5) The adhesion rate of the Lactobacillus curvature OH126 of this application to human vaginal epithelial cells VK2 / E6E7 is 19.8%-22.3%, which is higher than that of the control strain Lactobacillus acidophilus GLA-14 (13.5%-15.8%). It has a stronger potential for vaginal mucosal colonization and can competitively occupy epithelial cell adhesion sites to prevent pathogenic bacteria from invading.

[0043] The *Lactobacillus curvature* OH126 of this application can be formulated into lyophilized tablets or vaginal gel products for vaginal use in maintaining women's reproductive health. The preparation methods for the two formulations are as follows: (1) Preparation of lyophilized tablets for vaginal use: ①Cell collection: Lactobacillus curvature OH126 was inoculated into 10 L MRS liquid medium in a 20 L fermenter and anaerobically cultured at 37 °C for 13 h (until the late logarithmic growth phase, OD... 600 (Value 1.18-1.20), immediately after fermentation, centrifuge at 4℃ and 11000rpm for 18min, discard the supernatant and collect the sludge. The entire process is aseptic. The viable cell count in the sludge is approximately 1.0×10¹¹-1.2×10¹¹ CFU / g. ② Bacterial cell encapsulation: Mix the bacterial sludge with the freeze-drying protectant (12wt% skim milk powder, 8wt% trehalose, 5wt% sucrose, and sterile distilled water) at a mass ratio of 1:2, and stir slowly at 25℃ and 50rpm for 35 minutes to form a homogeneous bacterial suspension. ③ Freeze-drying: The bacterial suspension was dispensed into sterile freeze-drying molds at 0.5g / part, placed in a freeze dryer, pre-frozen at -45℃ for 3.5h, then heated to -25℃ at 5℃ / h and held for 2h, heated to -10℃ at 3℃ / h and held for 3h, and heated to 25℃ at 2℃ / h and held for 4h. The vacuum degree was 12-15Pa throughout the process, and the total freeze-drying time was 19h. The viable count of the bacterial blocks after freeze-drying was 2.0×10¹. 0 -2.5×10¹ 0 CFU / g; ④ Pulverization and mixing: The freeze-dried bacterial blocks are aseptically pulverized and passed through an 80-mesh sieve. They are then mixed according to the following mass proportions: 32 parts freeze-dried bacterial powder, 28 parts microcrystalline cellulose, 6 parts crospovidone, and 1.5 parts magnesium stearate. The mixture is stirred at 20 rpm for 30 minutes. ⑤ Tableting and Packaging: Tablets are formed using a rotary tablet press (model ZP-5) at a pressure of 22kN, with a diameter of 5mm and a thickness of 2mm. Each tablet contains ≥1×10⁻⁶ live bacteria. 9 CFU is packaged in aluminum-plastic blister packs, sealed and stored at 4°C, with a shelf life of 12 months.

[0044] (2) Preparation of vaginal gel: ① Preparation of enteric-coated microcapsules: Take the above-mentioned lyophilized bacterial powder, mix it with enteric encapsulation material (5wt% hydroxypropyl methylcellulose phthalate, 20wt% corn starch, 1wt% magnesium stearate), add sterile water to make a suspension, and spray dry (inlet air temperature 180℃, outlet air temperature 80℃) to prepare enteric-coated microcapsules. ② Gel matrix and mixing: Mix the enteric microcapsules with excipients (30wt% microcrystalline cellulose, 25wt% fructooligosaccharides, and 2wt% silica) evenly. Slowly add the gel matrix under aseptic conditions, stirring at 55rpm for 20min. Add purified water to a total mass of 100g and continue stirring for 10min to form a homogeneous gel. ③ Dispensing and sterilization: Dispense the gel into sterile pharmaceutical gel tubes, 5g per tube, and determine the viable count per tube is ≥1×10⁻⁶. 9 CFU is sterilized by 2.5kGy gamma irradiation, sealed and stored at 4°C, with a shelf life of 6 months.

[0045] The *Lactobacillus curvatureii* OH126 preparation of this application can be administered topically through the vagina to maintain vaginal microecological balance and improve problems related to vaginal microecological disorders. In vivo experiments in rats showed that the vaginal gel can effectively reduce the load of *Gardnerella vaginalis* in the vagina, repair vaginal mucosal damage, and reduce inflammatory cell infiltration, with effects comparable to 0.5% metronidazole gel without the risk of bacterial imbalance caused by antibacterial drugs; its bacterial suspension also showed preliminary adjuvant improvement effects on rat models of uterine fibroids, reducing fibroid volume and downregulating the expression of pro-angiogenic factor VEGF and pro-cell proliferation factor PCNA in fibroid tissue.

[0046] The present application will now be described in detail with reference to specific embodiments and accompanying drawings. These embodiments are for illustrative purposes only and should not be construed as limiting the scope of the application.

[0047] Example The culture medium involved in this example is as follows: MRS solid culture medium (g / L): peptone 10.0 g / L, beef extract 10.0 g / L, yeast extract 5.0 g / L, glucose 20.0 g / L, sodium acetate 5.0 g / L, diamine citrate 2.0 g / L, Tween-80 1.0 g / L, dipotassium hydrogen phosphate 0.4 g / L, magnesium sulfate 0.58 g / L, manganese sulfate 0.29 g / L, calcium carbonate 20.0 g / L, agar 15.0 g / L; autoclave at 121℃ for 20 min, cool and pour into plates for later use (for the isolation and culture of Lactobacillus curvature).

[0048] MRS liquid culture medium (g / L): peptone 10.0 g / L, beef extract 10.0 g / L, yeast extract 5.0 g / L, glucose 20.0 g / L, sodium acetate 5.0 g / L, diamine citrate 2.0 g / L, Tween-80 1.0 g / L, dipotassium hydrogen phosphate 0.4 g / L, magnesium sulfate 0.58 g / L, manganese sulfate 0.29 g / L, calcium carbonate 20.0 g / L; autoclaved at 121℃ for 20 min, cooled and used for the expansion culture of Lactobacillus curvature.

[0049] DMEM / F12 cell culture medium (g / L): 10.6 g / L DMEM / F12 basal medium powder, 100 mL / L fetal bovine serum (FBS), 10 mL / L penicillin-streptomycin mixture (final concentration of penicillin 100 U / mL, streptomycin 100 μg / mL); the basal medium was autoclaved at 121℃ for 20 min, and after cooling, FBS and penicillin were aseptically added for the culture of human vaginal epithelial cells VK2 / E6E7.

[0050] Columbia blood agar plates: used for the isolation and counting of Gardnerella vaginalis and Prevotella, the core pathogens of bacterial vaginosis.

[0051] Sabouraud dextrose agar plates: used for the culture and counting of Candida albicans, an opportunistic pathogen in the vagina.

[0052] Lactic acid assay kit: used to determine the lactic acid concentration in Lactobacillus curvature fermentation broth.

[0053] Experiment 1: Isolation, purification and identification of Lactobacillus curvatureii OH126 1. Isolation and purification of bacterial strains Lactobacillus curlis OH126 was isolated from vaginal secretion samples of healthy women of reproductive age. Specifically, the samples were obtained from healthy volunteers (aged 22-35) recruited by Dalian Medical University. Participants were required to have no history of reproductive system infections, organic diseases such as uterine fibroids, and had not used antibiotics, probiotic preparations, or hormone medications (including oral and topical medications) within the past 3 months. The samples were isolated during the construction of a vaginal flora bank and subsequently deposited at the Guangdong Provincial Center for Microbial Culture Collection.

[0054] (1) Pretreatment: Use sterile vaginal swabs to collect secretions from the middle and lower part of the vaginal wall. Take 0.1g of secretions and place the swab in a centrifuge tube containing 3mL of sterile PBS buffer (pH 7.2, with 1% penicillin-streptomycin antibiotics). Vortex for 10min to fully wash out the secretions and obtain the initial bacterial suspension. (2) Serial dilution: The initial bacterial suspension was serially diluted 10-fold with sterile PBS to prepare 10⁻¹, 10⁻², 10⁻³, and 10⁻¹ dilutions. 4 10⁻5 Concentration gradient bacterial suspensions; take 10⁻ 4 10⁻ 5 100 μL of bacterial suspension at each of the two dilutions; (3) Selective culture: The plates were dropped onto the surface of MRS solid medium and spread evenly in the horizontal and vertical directions using a sterile L-shaped glass rod. Three parallel plates were set up for each dilution. The plates were placed in an anaerobic workstation (gas composition: 85% nitrogen, 10% hydrogen, 5% carbon dioxide) and statically cultured at 37°C for 36 hours. The selective effect of MRS medium on lactobacillus was used to inhibit the growth of other bacteria. (4) Screening and purification of target colonies: After the culture is completed, observe the morphology of the plate colonies and screen the target colonies that meet the following characteristics: diameter 1.5-2.2 mm, round, milky white, smooth surface without wrinkles, neat edge, slightly raised center of the colony, and a clear and transparent calcium dissolution zone formed around it (with strong acid production ability); pick the above target colonies with a sterile inoculation loop and inoculate them into a sterile test tube containing 5 mL of MRS liquid medium and anaerobic culture at 37℃ for 24 h; take 1 mL of culture and inoculate it into MRS solid medium using the three-zone streak method and anaerobic culture at 37℃ for 48 h. Repeat the streak purification step 3 times. After each purification, take a single colony for Gram staining and microscopic examination to confirm that there are no rod-shaped, spherical or other contaminating bacteria morphologies and obtain pure culture strains; (5) Preservation of strains: The purified strains were inoculated into MRS liquid medium containing 20% ​​sterile glycerol, gently inverted and mixed, and then dispensed into 1.5 mL sterile cryovials, 1 mL per tube; they were pre-frozen at -20℃ for 2 h, and then transferred to an ultra-low temperature freezer at -80℃ for long-term cryopreservation; at the same time, they were inoculated into MRS solid plates and stored at 4℃ for a short period of time (not exceeding 1 month) for subsequent subculturing experiments.

[0055] 2. Identification of strains (1) Morphological identification: The purified bacterial strain was inoculated onto MRS solid plates and anaerobically cultured at 37°C for 48 hours. Colony morphology was then observed (e.g., ...). Figure 1 As shown): The colonies are round, 1.5-2.2 mm in diameter, with a smooth and glossy surface, neat edges without serrations, uniform texture, and a milky white to pale yellow color. When picked up, there are no filamentous adhesions, and the center of the colony is slightly raised. After Gram staining, a single colony smear is observed under a 1000× oil immersion optical microscope. The strain is Gram-positive, with short rod-shaped cells, no spores, and the cells are mostly arranged singly or in pairs.

[0056] (2) Determination of growth characteristics: Lactobacillus curvature OH126 and the control strain Lactobacillus acidophilus GLA-14 were inoculated into MRS liquid medium at an inoculum size of 2% (v / v) and cultured anaerobically at 37°C. Samples were taken at 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, and 24 h of culture, and the OD of the bacterial culture was measured using a UV spectrophotometer. 600 Values ​​(represented by solid lines, reflecting cell density) were analyzed. Simultaneously, 100 μL of bacterial suspension was serially diluted 10-fold, plated onto MRS agar plates, and viable cell counts were performed (CFU / mL, represented by dashed lines and dots). Growth curves were then plotted (see [link to MRS plate]). Figure 2 The horizontal axis represents the incubation time, and the vertical axis represents OD. 600 Value and viable count).

[0057] Growth curve analysis showed that *Lactobacillus curli* OH126 had a clear growth cycle, and its growth viability and stability were significantly better than the control strain *Lactobacillus acidophilus* GLA-14. During the lag phase (0-4 h), both strains gradually adapted to the environment; after entering the logarithmic growth phase (4-12 h), OH126 exhibited a faster proliferation rate, with a rapid increase in cell density and viable count, reaching a peak viable count of 3.60 × 10⁻⁶ at 12 h. 9 CFU / mL, compared to the peak value of 2.70 × 10⁻⁶ for GLA-14. 9 The CFU / mL was approximately 33% higher. Crucially, during the subsequent stationary phase (12-24 hours), OH126 exhibited superior physiological stability, maintaining a high viable count; while GLA-14, due to insufficient tolerance to the self-created acidic stress environment (such as low pH and high lactic acid concentration), showed a continuous decline in viable count, with a decrease of up to 37% by 24 hours. These results fully demonstrate that OH126 not only possesses rapid growth characteristics but also robust physiological functions to cope with environmental stress, providing a key strain advantage for achieving high yields in industrial high-density fermentation and long-term colonization in the acidic vaginal microecology.

[0058] (3) Molecular biological identification: 1) DNA extraction: Take 8 mL of bacterial culture in the logarithmic growth phase, centrifuge at 4℃ and 8000 rpm for 15 min, and discard the supernatant; then wash the bacterial cells three times with sterile PBS to remove residual components of the culture medium. Total DNA of the strain was extracted using a bacterial genomic DNA extraction kit (Hunan Aike Rui Biotechnology Co., Ltd., catalog number AG21007); the purity and integrity of the DNA were verified by 1% agarose gel electrophoresis.

[0059] 2) PCR amplification: Using the extracted DNA as a template, PCR amplification was performed using Lactobacillus-specific primers. The upstream primer Lac-F (sequence: 5'-AGCAGTAGGGAATCTTCCA-3') (Seq ID No. 1) and the downstream primer Lac-R (sequence: 5'-ATTYCACCGCTACACATGG-3') (Seq ID No. 2) were synthesized by Shanghai Sangon Biotech Co., Ltd. The PCR reaction system was 50 μL, containing 2 μL each of the upstream and downstream primers (10 μmol / L), 3 μL of DNA template, and the remainder was premixed solution (Wuhan Aibotek Biotechnology Co., Ltd., catalog number RK20607). The reaction conditions were: 95℃ pre-denaturation for 5 min; 95℃ denaturation for 30 s, 56℃ annealing for 40 s, 72℃ extension for 1 min, for a total of 35 cycles; and 72℃ final extension for 10 min.

[0060] 3) Sequencing and alignment: The PCR products were sequenced by Wuhan Aijibaike Biotechnology Co., Ltd., and the 16S rRNA gene sequencing results of Lactobacillus curvature OH126 were obtained (specific sequence: >06H+D+1500+...+a2306266743-RS-MRS-126-Contig1) Seq ID No.:

[0061]

[0062] Experiment 2: Determination of the functional characteristics of Lactobacillus curvature OH126 1. Detection of lactic acid production capacity of Lactobacillus curvature OH126 After activating *Lactobacillus curvature* OH126 to the logarithmic developmental phase, it was inoculated into MRS liquid medium at a 2% (v / v) inoculum and cultured in an anaerobic incubator at 37°C for 36 h. Simultaneously, *Lactobacillus acidophilus* GLA-14 was used as a control strain and cultured under the same inoculum and conditions. After culture, 10 mL of fermentation broth was centrifuged at 4°C and 6000 rpm for 20 min. The supernatant was collected and filtered through a 0.22 μm sterile filter to remove bacterial cells and culture medium impurities. The lactic acid concentration in the supernatant was determined using a lactic acid assay kit, and the pH of the fermentation broth was directly measured using a precision pH meter to assess the strain's ability to construct an acidic vaginal microenvironment through acid production.

[0063] The results of the lactic acid production capacity assays for *Lactobacillus curvatureii* OH126 and *Lactobacillus acidophilus* GLA-14 in this example are shown in Table 1.

[0064] Table 1 Results of lactic acid production capacity assay Detection strain Lactic acid production (g / L) Fermentation broth pH value Lactobacillus crispatus OH126 12.5-14.0 3.6-3.8 Lactobacillus acidophilus GLA-14 9.1-9.9 4.1-4.4 2. Detection of hydrogen peroxide production capacity of Lactobacillus curvature OH126 MRS solid plates containing 0.25 mg / mL TMB and 0.01 mg / mL horseradish peroxidase were prepared. *Lactobacillus curvatureii* OH126 was cultured to the logarithmic growth phase, and the bacterial concentration was adjusted to 1 × 10⁻⁶ with sterile physiological saline. 8 CFU / mL, take 100 μL and spread it evenly on the surface of the plate. After anaerobic incubation at 37℃ for 48 h, take it out, expose it to air and time it, and record the time when the colony changes from colorless to blue. At the same time, use Lactobacillus acidophilus GLA-14 as a control strain and treat it with the same steps to evaluate the difference in hydrogen peroxide production ability between the two strains.

[0065] The results of the hydrogen peroxide production capacity determination of Lactobacillus curvature OH126 and Lactobacillus acidophilus GLA-14 in this example are shown in Table 2.

[0066] Table 2 Results of hydrogen peroxide production capacity measurement Strain Discoloration time after exposure to air (min) Hydrogen peroxide production capacity Lactobacillus crispatus OH126 16-18 Moderate (slightly strong) Lactobacillus acidophilus GLA-14 21-23 Moderate (slightly weak) 3. Antibacterial activity test of Lactobacillus curvature OH126 The inhibitory effect of *Lactobacillus curvatureii* OH126 on common vaginal pathogens was evaluated using a 96-well plate gradient dilution combined with CFU counting. The target pathogens included *Gardnerella vaginalis* (ATCC 14018), *Prevotella bivia* (ATCC 29303), *Mobiluncus mulieris* (ATCC 35243), and *Candida albicans* (ATCC 10231).

[0067] The specific steps are as follows: In a 96-well plate, an equal amount of the above four pathogenic bacteria solutions (1×10⁶ bacteria per well) was first added to each well. 5 CFU were then added to different proportions (10%, 20%, 30%) of *Lactobacillus crinoides* OH126 culture supernatant to bring the total volume of each well to 200 μL. MRS medium was used as a positive control instead of the *Lactobacillus* supernatant, and a 30% supernatant treatment group of *Lactobacillus acidophilus* GLA-14 was set up as a reference. After incubating the 96-well plate at 37°C for 12 hours, the CFU counting method was used: each group of culture was serially diluted 10-fold, and an appropriate amount of the dilution was spotted onto the corresponding solid medium (Columbia blood agar plates for *Gardnerella vaginalis* and *Prevotella*, MRS solid medium for *Morchella molybdenum*, and Sabouraud dextrose agar plates for *Candida albicans*). Statistical analysis was performed after the colonies grew to a suitable size for counting.

[0068] The results are as follows Figure 3 As shown, *Lactobacillus curvatureii* OH126 exhibited inhibitory effects against all four selected vaginal pathogens. With the concentration of the OH126 culture supernatant increasing from 10% to 30%, its inhibitory effect on each pathogen gradually strengthened. Furthermore, at a 30% supernatant concentration, OH126 showed superior inhibitory effects against all four pathogens compared to the 30% supernatant treatment group of *Lactobacillus acidophilus* GLA-14. For example, against *Gardnerella vaginalis*, the viable bacterial count in the OH126 30% supernatant group was only about 30% of the positive control, while the viable bacterial count in the GLA-14 30% supernatant group was about 55% of the positive control; against *Candida albicans*, the viable bacterial count in the OH126 30% supernatant group was about 25% of the positive control, while the GLA-14 30% supernatant group was about 50%. These results clearly demonstrate that *Lactobacillus curvatureii* OH126 has significant advantages in antibacterial function, providing direct experimental evidence for its application in the field of women's reproductive health.

[0069] 4. In vitro antibacterial experiment of Lactobacillus curvature OH126 supernatant While confirming the inhibitory effect of *Lactobacillus curvatureii* OH126 supernatant on common vaginal pathogens, the effect of OH126 supernatant on host cell viability was investigated to further verify its safety. The host cells were RAW264.7 mouse macrophages (purchased from the Cell Bank of the Chinese Academy of Sciences, cultured under DMEM + 10% FBS conditions). To evaluate the safety of *Lactobacillus curvatureii* OH126 supernatant on host cells, the CCK-8 assay was used to detect cell viability at different supernatant concentrations (1, 10, 20, 30, 40 μg).

[0070] The specific method was as follows: *Lactobacillus curvatureii* OH126 in the logarithmic growth phase was cultured, and its fermentation supernatant was collected and sterilized by filtration. Supernatants of different concentrations were added to the cell culture system, and after 24 hours of incubation, CCK-8 reagent was added. The CCK-8 reagent was diluted 1:10 with serum-free DMEM, and 10 μL of the diluted CCK-8 solution was added to each well. After incubation at 37℃ and 5% CO2 for 2 hours, cell viability was detected by measuring the absorbance at 450 nm using a microplate reader. The results are as follows: Figure 4 As shown, the horizontal axis represents the amount of OH126 supernatant loaded (μg / well), with five concentration gradients of 1, 10, 20, 30, and 40 μg / well. The vertical axis represents the RAW264.7 cell viability (%), calculated with the survival rate of the blank control group (containing only cells and culture medium) being 100%.

[0071] The results showed that cell viability remained at a high level (>85%) in all treatment groups, with the 1 μg group at 100.0% ± 5.0%, the 10 μg group at 95.0% ± 6.0%, the 20 μg group at 93.0% ± 7.0%, the 30 μg group at 88.0% ± 5.0%, and the 40 μg group at 90.0% ± 6.0%. This indicates that the supernatant of *Lactobacillus crenulata* OH126 exhibits significant antibacterial activity while showing no obvious toxicity to host cells, demonstrating good safety (see...). Figure 4 ).

[0072] Experiment 3: Determination of the tolerance of Lactobacillus curvature OH126 to the vaginal environment 1. Acid resistance test of Lactobacillus curvature OH126 MRS liquid culture media with pH values ​​of 3.0, 3.5, 4.0, 4.5, and 5.0 were prepared (pH value was adjusted with 1 mol / L hydrochloric acid or 1 mol / L sodium hydroxide, and sterilized by filtration through a 0.22 μm filter membrane) to simulate the difference in acidic environment between a healthy vagina (pH 3.8-4.5) and a patient with bacterial vaginosis (pH > 4.6).

[0073] After taking OH126 bacterial suspension in the logarithmic growth phase and determining the initial viable count (N0), inoculate it into culture media of different pH values ​​at a 1:10 ratio, incubate at 37°C for 2 hours, and then determine the viable count (N0) again.t ), according to "Survival rate = (N t The survival rate was calculated as (N0) × 100%. Lactobacillus acidophilus GLA-14 was used as a control strain, and experiments were conducted simultaneously.

[0074] 2. Bile salt tolerance test of Lactobacillus curvature OH126 Based on the possible bile salt concentrations in vaginal secretions, a gradient of sodium taurocholate (0.1%, 0.2%, 0.3%, 0.4%, and 0.5% (w / v)) was established and added to MRS medium (sterilized by filter membrane). Logarithmic-phase OH126 bacterial suspension was taken, and after determining the initial viable count, it was inoculated at a 1:10 ratio into mediums containing different bile salt concentrations. The mixture was incubated at 37°C for 4 hours, and the viable count was determined and the survival rate calculated. Lactobacillus acidophilus GLA-14 was used as a control. Three parallel experiments were conducted, denoted as Rep1, Rep2, and Rep3.

[0075] The results are as follows Figure 5 As shown, under two simulated vaginal environmental stress conditions, the survival rates of both *Lactobacillus curvatureii* OH126 and the control strain *Lactobacillus acidophilus* GLA-14 gradually decreased with increasing stress factor concentrations (acidity intensity and bile salt concentration), but OH126 showed better tolerance. Within the common acidic vaginal environment range (pH 3.5-4.5), OH126 maintained a high survival rate, and even under strongly acidic conditions at pH 3.0, its survival ability was still superior to GLA-14. Furthermore, within the common bile salt concentration range of 0.1%-0.3% in vaginal secretions, OH126 also maintained a high survival rate, and even when the bile salt concentration rose to 0.4% or higher, its survival performance was better than the control strain.

[0076] Overall, Lactobacillus curvature OH126 exhibits good adaptability to the acidic conditions and bile salt stimulation in the vaginal microenvironment, providing an environmental tolerance basis for its stable colonization in the vagina and the exertion of its probiotic effects.

[0077] Experiment 4: Determination of the adhesion ability of Lactobacillus curvature OH126 to vaginal epithelial cells Adhesion ability is a core characteristic for vaginal probiotics to colonize and exert their effects on the mucosal surface. For *Lactobacillus curvature*, its ability to stably adhere to vaginal epithelial cells directly determines its ability to compete with pathogens for binding sites, prevent pathogens from invading the mucosal barrier, and thus maintain the vaginal microecological balance. This embodiment evaluates the adhesion ability of *Lactobacillus curvature* OH126 to vaginal epithelial cells through in vitro cell experiments, using *Lactobacillus acidophilus* GLA-14, a commonly used clinical vaginal probiotic, as a control. The specific procedures are as follows: Human vaginal epithelial cells VK2 / E6E7 were purchased from the Cell Bank of the Chinese Academy of Sciences. They were cultured in DMEM / F12 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin mixture, and placed in a 37°C, 5% CO2 incubator. When the cells reached approximately 80% confluence in the culture flasks, the old medium was discarded, and 1 mL of 0.25% trypsin-EDTA solution was added. The cells were anaerobically incubated at 37°C for 2 minutes to detach from the flask walls, and then gently pipetted to prepare a single-cell suspension. 3 mL of fresh DMEM / F12 medium was added to the suspension to stop digestion, and the mixture was stirred to adjust the cell concentration to 4 × 10⁶ cells / mL. 4 Take 2 mL of cell suspension and seed it into a 6-well cell culture plate. Continue culturing for 24 hours until the cells form a dense and uniform monolayer.

[0078] Lactobacillus curvature OH126 and Lactobacillus acidophilus GLA-14 were inoculated separately into MRS liquid medium and cultured anaerobically at 37°C until the logarithmic growth phase. 5 mL of each strain was centrifuged at 3000 rpm for 10 minutes at 4°C, and the supernatant was discarded to remove culture medium components. The bacterial pellet was resuspended in sterile PBS, and the centrifugation and washing were repeated three times to thoroughly remove residual culture medium. Finally, the bacterial suspension was resuspended in sterile PBS to prepare a bacterial suspension. A small amount of the bacterial suspension was spread onto MRS agar plates and anaerobically cultured at 37°C for 48 hours. The bacterial counts were then performed to ensure that the concentration of both strains in the suspension was adjusted to 1 × 10⁻⁶. 8 CFU / mL, this value is recorded as "pre-adhesion viable bacteria count".

[0079] Before starting the adhesion experiment, remove the DMEM / F12 medium from the 6-well plate, gently wash the cell monolayer three times with 2 mL of sterile PBS to remove any unattached suspended cells, add 2 mL of the prepared bacterial suspension to each well (3 parallel wells for both the OH126 and GLA-14 groups), gently shake the plate to evenly cover the cell surface with the bacterial suspension, and incubate at 37°C, 5% CO2 for 2.5 hours to allow the bacteria to fully contact and adhere to the cells. After incubation, carefully aspirate the unattached bacterial suspension from the wells and gently wash the cells three times with sterile PBS to avoid interference from residual free bacteria during counting. Add 0.5 mL of 0.25% trypsin-EDTA solution to each well and incubate at 37°C for 5 minutes to digest the cells. Once the cells have completely detached from the cell wall, add 1 mL of fresh DMEM / F12 medium to stop the digestion. Gently pipette the cells to detach them from the surface bacteria. Then add 1 mL of 0.05% Triton X-100 solution and incubate at room temperature for 10 minutes to lyse the cells, releasing any remaining intracellular and surface bacteria to obtain a lysate containing the adhered bacteria.

[0080] Take 100 μL of lysis buffer and perform 10-fold serial dilutions with sterile PBS, selecting an appropriate dilution (usually 10⁻³-10⁻). 4 Take 100 μL of the diluted solution and spread it on an MRS solid plate. After anaerobic incubation at 37°C for 48 hours, count the number of colonies. This value is recorded as "the number of viable bacteria after adhesion".

[0081] The adhesion rates of the two strains were calculated using the formula "Adhesion rate = (Number of viable bacteria after adhesion / Number of viable bacteria before adhesion) × 100%", and the results are shown in Table 3. Table 3. Results of the adhesion experiment of vaginal epithelial cells of Lactobacillus curvature OH126 Strain Adhesion rate to human vaginal epithelial cells VK2 / E6E7 Lactobacillus crispatus OH126 19.8%-22.3% Lactobacillus acidophilus GLA-14 13.5%-15.8% As clearly shown in Table 3, *Lactobacillus curvatureii* OH126 exhibited a significantly higher adhesion rate to human vaginal epithelial cells VK2 / E6E7 than the control strain *Lactobacillus acidophilus* GLA-14, indicating that OH126 possesses stronger vaginal mucosal colonization potential. In practical applications, this characteristic helps OH126 to remain more stably in the vagina, competitively occupying adhesion sites on the surface of epithelial cells and reducing the chances of pathogenic bacteria such as *Gardnerella vaginalis* and *Candida albicans* binding to cells. This provides strong support for inhibiting the proliferation of pathogenic bacteria and maintaining vaginal microecological stability.

[0082] Experiment 5: Determination of the biochemical characteristics of Lactobacillus curvature OH126 1. Catalase experiment Pick a fresh single colony of OH126 cultured for 48 hours on an MRS solid plate, place it in the center of a clean glass slide, add 2 drops of 0.3% hydrogen peroxide solution, and observe the phenomenon within 1 minute. The immediate appearance of a large number of bubbles indicates a positive (+) result, while the absence of bubbles indicates a negative (-) result.

[0083] 2. Gelatin liquefaction experiment Prepare gelatin culture medium (10 g / L tryptone, 120 g / L gelatin, 1000 mL distilled water, pH 7.2), autoclave at 121 °C for 20 min, cool, dispense, and allow to solidify. Use a sterile inoculation needle to pick up a single colony of OH126 and vertically puncture it into the culture medium (to a depth of 4 / 5 of the medium's height). Incubate anaerobically at 28 °C for 7 days, then refrigerate at 4 °C for 30 min. Observe the medium; a positive (+) result indicates partial or complete gelatin solidification, while a negative (-) result indicates complete gelatin solidification.

[0084] 3. Hydrogen sulfide experiment Take lead acetate medium, pick a single colony of OH126 with a sterile inoculation needle, and inoculate it to the bottom of the medium. Incubate anaerobically at 37°C for 48 hours. Observe whether black precipitate appears. If the medium turns black, it is a positive (+) result; if the medium does not change color, it is a negative (-) result.

[0085] 4. Nitrate Reduction Experiment Prepare nitrate medium (10 g / L peptone, 0.5 g / L potassium nitrate, 1000 mL distilled water, pH 7.2), sterilize and dispense into individual vials, inoculate with OH126, and anaerobic incubate at 37°C for 3 days. Take 1 mL of the culture medium and add 0.1 mL each of nitrate reducing reagent A and B, and incubate at room temperature for 5 minutes. Observe the color change of the liquid in the test tube; a red color indicates a positive (+) result, while no color reaction after adding the reagents indicates a negative (-) result.

[0086] The biochemical reaction tests of Lactobacillus curvature OH126 are shown in Table 1.

[0087] Table 4 Biochemical reaction tests of Lactobacillus curvature OH126 Strain Test of catalase Gelatin liquefaction test Hydrogen sulfide test Nitrate reduction test Lactobacillus crispatus OH126 - - - - The results in Table 4 show that Lactobacillus curlis OH126 does not contain catalase, cannot liquefy gelatin, does not produce hydrogen sulfide, and does not reduce nitrates, which is consistent with the typical biochemical characteristics of Lactobacillus curlis.

[0088] Experiment 6. Preparation of Lactobacillus curvatureii OH126 inoculum 1. Preparation of Lyophilized Tablets for Vaginal Use of Lactobacillus curvature OH126 (1) Cell collection: Lactobacillus curvature OH126 was inoculated into 10 LMRS liquid medium (in a 20 L fermenter) and anaerobically cultured at 37℃ for 13 h (until the late logarithmic growth phase, OD 600 (Value 2.8-3.0); After fermentation, immediately centrifuge at 11000rpm for 18min at 4℃, discard the supernatant, and collect the bacterial precipitate (bacterial sludge). Aseptic operation is performed throughout the process to ensure the purity of the bacterial cells (no contaminants under microscopic examination); the number of viable bacteria in the bacterial sludge is determined to be approximately 1.0×10¹¹-1.2×10¹¹CFU / g.

[0089] (2) Encapsulation of bacterial cells: Add freeze-drying protectant solution (formulation: 12wt% skim milk powder, 8wt% trehalose, 5wt% sucrose, prepared with sterile distilled water) to the bacterial sludge at a mass ratio of 1:2. Stir slowly at 50 rpm for 35 min at 25℃ to make the bacterial cells evenly dispersed in the protectant solution and form a uniform bacterial suspension.

[0090] (3) Freeze-drying: The bacterial suspension was dispensed into sterile freeze-drying molds, 0.5 g per mold; the molds were placed in a freeze dryer, and the freeze-drying parameters were set as follows: first, pre-freeze at -45℃ for 3.5 h; then, raise the temperature to -25℃ at a rate of 5℃ / h and hold for 2 h; then raise the temperature to -10℃ at a rate of 3℃ / h and hold for 3 h; finally, raise the temperature to 25℃ at a rate of 2℃ / h and hold for 4 h; the vacuum degree was maintained at 12-15 Pa during the freeze-drying process, and the total freeze-drying time was 19 h; after the freeze-drying was completed, the freeze-dried bacterial blocks were taken out and the viable count was measured, which reached 2.0 × 10¹. 0 -2.5×10¹ 0 CFU / g.

[0091] (4) Crushing and mixing: Crushing the freeze-dried bacterial blocks with a sterile crusher and passing them through an 80-mesh sterile sieve to obtain freeze-dried bacterial powder with uniform particle size; Weigh out the following by mass: 32 parts freeze-dried bacterial powder, 28 parts microcrystalline cellulose (filler), 6 parts cross-linked polyvinyl chloride (disintegrant), and 1.5 parts magnesium stearate (lubricant), place them in a sterile mixer, and mix at a rate of 20 rpm for 30 min to ensure uniform mixing.

[0092] (5) Tableting and Packaging: The mixture is added to a rotary tablet press (Shanghai Tianfan Pharmaceutical Machinery Manufacturing Plant, model ZP-5), and the tableting pressure is set to 22kN to compress it into vaginal lyophilized tablets with a diameter of 5mm and a thickness of 2mm; the viable count of each tablet is measured to ensure ≥1×10 9 CFU / tablet; packaged in aluminum-plastic blister packs with oxygen barrier layer (10 tablets per blister pack), and stored in a sealed container at 4°C.

[0093] 2. Preparation of Lactobacillus curvatureii OH126 vaginal gel (1) Preparation of bacterial suspension: Collect Lactobacillus curvature OH126 bacterial sludge according to the above steps, add sterile physiological saline, stir evenly with a sterile homogenizer, and adjust the bacterial suspension concentration to 1.5×10¹ 0 CFU / mL was prepared as a bacterial culture for later use.

[0094] (2) Gel matrix preparation: Take 1.8g of carbomer 940 (gel matrix), add 85mL of purified water, and place at room temperature for 24h. Stir once every 6h (at a speed of 30rpm) to allow the carbomer to fully swell and form a uniform and transparent gel matrix. Add 6g of glycerin (humectant) and 0.4g of phenoxyethanol (preservative) to the matrix, stir evenly, and adjust the pH of the gel to 4.0 with 10% citric acid solution to simulate the pH environment of a healthy vagina.

[0095] (3) Mixing and dispensing: Under aseptic conditions, the prepared bacterial solution was slowly added to the gel matrix at a rate of 55 rpm while stirring for 20 minutes to ensure that the bacterial solution was evenly dispersed in the gel; purified water was added to a total mass of 100 g, and stirring was continued for 10 minutes to form a homogeneous vaginal gel; the gel was dispensed into sterile pharmaceutical gel tubes, 5 g per tube, and the viable bacteria count in each tube was determined to be ≥1×10⁻⁶. 9 CFU / branch.

[0096] (4) Sterilization and preservation: The dispensed gel tubes are sterilized by γ-ray irradiation (dose 2.5kGy to ensure no contamination by other bacteria and to ensure no impact on bacterial activity); after sterilization, they are sealed and stored at 4℃, with a shelf life of up to 6 months.

[0097] Experiment 7: In vivo efficacy verification experiment of Lactobacillus curvatureii OH126 vaginal gel Forty SPF-grade female SD rats (weighing 180-200g, 6 weeks old) were selected and randomly divided into four groups after one week of acclimatization: healthy group, model group, metronidazole group, and OH126 gel group, with 10 rats in each group. Except for the healthy group, the other three groups were vaginally inoculated with Gardnerella vaginalis suspension (concentration 1×10⁻⁶). 8 A vaginal microecological disorder model was constructed by inoculating each rat with CFU / mL (0.1 mL per rat per injection) for 3 consecutive days. On the 4th day of modeling, vaginal secretions from 3 rats in the model group were randomly selected for plate counting, confirming that the Gardnerella vaginalis load was significantly higher than that in the healthy group (up to 10). 6 The rat model was considered successfully established when the rats showed slight redness and swelling in their vaginas (at the CFU / mL level).

[0098] All interventions were conducted daily in the evening, using sterile vaginal applicators for precise administration: 0.1 mL of sterile saline was injected into each healthy group and model group per dose; 0.1 mL of 0.5% metronidazole gel was injected into each vagina group per dose, serving as a clinically used antibacterial control; and 0.1 mL of vaginal gel containing the antibacterial agent from Example 6 (with ≥1×10⁻⁶ viable OH126 bacteria in the gel) was injected into each vagina group per dose. 9 (CFU / g). Each group was administered the drug for 7 consecutive days. During the administration period, the rats' activity status, food intake, and vaginal appearance were recorded daily to monitor for adverse reactions such as mucosal damage and abnormal secretions.

[0099] After administration, all rats were euthanized by cervical dislocation, and vaginal tissue and secretions were rapidly dissected for testing of three core indicators: (1) HE staining and inflammation scoring of vaginal tissue: Vaginal tissue was fixed with 4% paraformaldehyde for 24 hours, dehydrated, embedded in paraffin, and then prepared into 5μm sections for HE staining; the mucosal condition was observed under an optical microscope, and the degree of inflammation was quantified by a three-level scoring standard of "mucosal redness-damage-inflammatory cell infiltration": ① Mucosal redness: no redness (0 points), mild redness (1 point), moderate redness (2 points), severe redness (3 points); ② Mucosal damage: no damage (0 points), local punctate damage (1 point), local patchy damage (2 points), large area damage (3 points); ③ Inflammatory cell infiltration: no inflammatory cells (0 points), a few scattered (1 point), moderate aggregation (2 points), large aggregation (3 points); The total inflammation score is the sum of the scores of the three items (full score 9 points).

[0100] (2) Gardnerella vaginalis count: After serial dilution of vaginal secretions with sterile physiological saline, 100 μL of the diluted solution was spread on Columbia blood agar plates and anaerobically cultured at 37°C for 48 hours. The number of Gardnerella vaginalis colonies was counted and the load was calculated.

[0101] (3) Vaginal pH value measurement: The pH value of vaginal secretions was directly measured using precision pH test paper (range 3.0-5.0) to assess the recovery of the acidic microenvironment.

[0102] The test results are shown in Table 5. Figure 6 A comparison chart of vaginal mucosal inflammation scores in different groups of rats.

[0103] Table 5. Effects of Lactobacillus curvatureii OH126 vaginal gel on a rat model of vaginal microecological disorder (x±s, n=10) Group Vaginal mucosa state (observed by HE staining) Vaginal pH value Inflammation score (total score 9) Gardnerella load Healthy group Mucosa smooth, no redness, almost no inflammatory cells 3.8±0.1 0.3±0.2 0.3 ± 0.1 x 10 6 CFU / mL Model group Mucosa obviously red and swollen, with damage and many inflammatory cells 4.7±0.2 7.8±0.6 8.2 ± 0.6 x 10 6 CFU / mL Metronidazole group Mucosa slightly red and swollen, with reduced damage and fewer inflammatory cells 4.1±0.1 3.2±0.4 3.5 ± 0.4 x 10 6 CFU / mL OH126 gel group Mucosa basically smooth, no obvious redness and damage, and very few inflammatory cells 3.9±0.1 2.9±0.3 3.2 ± 0.3 x 10 6 CFU / mL Figure 6 The chart is a bar graph, with the horizontal axis representing the healthy group, model group, metronidazole group, and OH126 gel group, respectively, and the vertical axis representing the vaginal mucosal inflammation score (unit: points, full score 9 points). Data in the graph are expressed as mean ± standard deviation, and error bars represent the range of data fluctuation within each group. As can be seen from the graph, the inflammation score in the model group was significantly higher than the other three groups. The inflammation scores in both the OH126 gel group and the metronidazole group were significantly lower, and there was no statistically significant difference between the two groups (P>0.05), clearly demonstrating the anti-inflammatory effect of OH126 gel.

[0104] From Table 5 and Figure 6 It was found that the model group rats exhibited significant vaginal mucosal damage, high inflammation scores, high pathogenic bacterial loads, and elevated pH levels, demonstrating the successful establishment of the microecological dysbiosis model. Compared to the model group, the OH126 gel group showed a decrease in vaginal mucosal inflammation score to 2.9±0.3 points, significant restoration of epithelial integrity, and a reduction in Gardnerella vaginalis bacterial load to (3.2±0.3)×10⁻⁶. 6The concentration of CFU / mL was comparable to that of the metronidazole group (P>0.05). This indicates that Lactobacillus curvaturei OH126 vaginal gel can not only effectively inhibit the proliferation of pathogenic bacteria in the vagina and repair mucosal damage, but also restore the acidic microenvironment of the vagina, while avoiding the risk of vaginal flora imbalance that may be caused by antibacterial drugs. It has good safety and practicality in the regulation of vaginal microecology.

[0105] Experiment 8: A preliminary study on the use of Lactobacillus curvature OH126 to assist in the improvement of uterine fibroids. Forty SPF-grade female SD rats (weighing 200-220g, 6 weeks old) were randomly divided into four groups: healthy group, model group, saline group, and OH126 bacterial suspension group, with 10 rats in each group. Except for the healthy group, the other three groups were used to establish a uterine fibroid model by subcutaneous injection of estradiol benzoate: injections were given twice a week at a dose of 0.5 mg / kg each time for 8 consecutive weeks. At the end of the 8th week of modeling, two rats in the model group were randomly sacrificed, and the uterine tissue was dissected, fixed in 4% paraformaldehyde, embedded in paraffin, stained with hematoxylin and eosin (HE), and observed under a light microscope for the presence of fibroid nodules in the myometrium.

[0106] The OH126 bacterial suspension group was administered 0.2 mL of a bacterial suspension diluted with the bacterial agent from Example 6 (live bacteria concentration 1×10¹) via a sterile vaginal applicator every evening. 0 The saline group received an equal volume of sterile saline solution vaginally every evening; the healthy group received no intervention and was fed only routinely (free access to food and water, ambient temperature 22-25℃, humidity 50%-60%). All groups underwent intervention for 4 consecutive weeks. During the intervention period, rats were weighed at the same time each week, and their mental state and reproductive system appearance were observed to exclude the influence of non-experimental factors.

[0107] After the intervention, all rats were euthanized and dissected. ① Remove the uterine tissue and accurately measure the long axis (a) and short axis (b) of the fibroid nodules with calipers. Calculate the average volume of each group of fibroids according to the formula "fibroid volume = 0.5 × a × b²". ② Take 0.1g of fibroid tissue, add RIPA lysis buffer containing protease inhibitor (1mL of lysis buffer per 100mg of tissue), homogenize thoroughly with a homogenizer under ice bath conditions, centrifuge at 12000rpm for 15 minutes at 4℃, and collect the supernatant as protein extract. ③ The expression levels of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) in the protein extract were detected by Western blot. β-actin was used as an internal reference protein. The gray values ​​of the protein bands were quantitatively analyzed by ImageJ software, and the relative expression level of the target protein (target protein gray value / β-actin gray value) was calculated.

[0108] The test results are shown in Table 6: Table 6. The auxiliary improvement effect of Lactobacillus curvature OH126 on rat uterine fibroid model (x±s, n=8-10) Group Myoma volume (cm³) Relative expression amount of VEGF Relative expression amount of PCNA Healthy group No myoma formation - - Model group 3.5±0.5 0.95±0.10 0.92±0.09 Saline group 3.3±0.4 0.93±0.09 0.89±0.08 OH126 bacterial suspension group 1.8±0.3 0.62±0.08 0.58±0.06 As shown in Table 6, no fibroids formed in the healthy group, while the fibroids in the model group and the saline group were larger and had higher levels of VEGF and PCNA expression, demonstrating the successful establishment of the uterine fibroid model and the lack of improvement effect of saline. Compared with the above two groups, the fibroid volume in the OH126 bacterial suspension group was significantly reduced, and the relative expression levels of VEGF (angiogenic factor) and PCNA (cell proliferation factor) were both decreased. This suggests that *Lactobacillus curvatureis* OH126 may inhibit the nutrient supply and excessive cell proliferation of fibroids by downregulating the expression of pro-angiogenic and pro-cell proliferation-related factors in fibroid tissue, thereby helping to delay fibroid growth. This provides preliminary experimental evidence for its subsequent research and application in improving uterine fibroid-related symptoms.

[0109] In summary, this study systematically investigated *Lactobacillus curlis* OH126. This strain was isolated from vaginal secretions of healthy women of childbearing age. Morphological, growth characteristic, and molecular biological identification (99% homology with *Lactobacillus curlis*) confirmed accurate classification. Four biochemical tests, including catalase and gelatin liquefaction tests, were all negative, consistent with its typical characteristics and without risk of contamination by other bacteria. Functionally, this strain is highly adapted to the needs of the vaginal microecology: lactic acid production is 12.5-14.0 g / L (fermentation broth pH 3.6-3.8), hydrogen peroxide production is superior to *Lactobacillus acidophilus* GLA-14, it inhibits four vaginal pathogens including *Gardnerella vaginalis*, and it exhibits high survival rates under common vaginal pH conditions of 3.5-4.5 and bile salt concentrations of 0.1%-0.3%. Vaginal epithelial cell adhesion rates are 19.8%-22.3% (significantly higher than GLA-14), facilitating stable colonization. The preparation process for the bacterial agent is mature, and vaginal freeze-dried tablets (≥1×10⁶ tablets per tablet) can be produced. 9 CFU (stored at 4°C for 12 months) and gel (each vial ≥ 1×10⁻⁶ CFU, ... 9 CFU (radiation sterilization safe), suitable for local application scenarios. In vivo experiments confirmed that its gel can improve vaginal microecological dysbiosis in rats (with effects similar to metronidazole but without bacterial imbalance), and the bacterial suspension can reduce the volume of rat fibroids and downregulate VEGF / PCNA expression. In summary, the functional characteristics of this strain are well-suited to the needs of vaginal microecology, the preparation process of the bacterial agent has been preliminarily established and is feasible, and in vivo experiments have confirmed its good safety and its effect in assisting the improvement of uterine fibroids. This indicates that *Lactobacillus curvatureii* OH126 has clear development value and application prospects in the field of women's reproductive health, especially in the microecological auxiliary intervention of uterine fibroids.

[0110] The above description, in conjunction with specific embodiments, provides a detailed explanation of this application and should not be construed as limiting the implementation of this application to these descriptions. For those skilled in the art, several simple deductions or substitutions can be made without departing from the basic inventive concept of this application.

Claims

1. A strain of *Lactobacillus curvature*, characterized by: The *Lactobacillus curvature* is *Lactobacillus curvature* OH126, GDMCC No. 66536.

2. A microbial agent, characterized in that: The microbial agent contains *Lactobacillus curvaturei* OH126 as described in claim 1; the active ingredient of the microbial agent is selected from at least one of live *Lactobacillus curvaturei* OH126 cells, *Lactobacillus curvaturei* OH126 culture medium, or *Lactobacillus curvaturei* OH126-derived fermentation products.

3. The microbial agent according to claim 2, characterized in that: The dosage form of the bacterial agent is a lyophilized tablet or vaginal gel for vaginal use; each unit of the bacterial agent contains ≥1×10⁻⁶ live Lactobacillus OH126 bacteria. 9 CFU, where vaginal lyophilized tablets are measured in "tablets" and vaginal gels are measured in "tubes".

4. The method for preparing the microbial agent according to claim 2 or 3, characterized in that: When the antifungal agent is in the form of a vaginal lyophilized tablet, the following steps are included: (1) Collection of bacterial cells: The Lactobacillus curvature OH126 described in claim 1 was inoculated into MRS liquid culture medium and cultured anaerobically at 37°C for 12-14 h until the late logarithmic growth phase. Then, it was centrifuged at 10000-12000 rpm for 15-20 min at 4°C. The supernatant was discarded and the live bacterial cells were collected. The entire process was carried out under aseptic conditions. (2) Cell encapsulation: Add freeze-drying protectant to the cells collected in step (1) and mix well; (3) Freeze-drying: The bacterial cells with added freeze-drying protectant were dispensed into freeze-drying molds and placed in a freeze dryer for freeze-drying. First, they were pre-frozen at -45℃ for 3-4 hours, then heated to -25℃ at a rate of 5℃ / h and held for 2 hours, then heated to -10℃ at a rate of 3℃ / h and held for 3 hours, and finally heated to 25℃ at a rate of 2℃ / h and held for 4 hours. The total freeze-drying time was 18-20 hours. The vacuum degree was maintained at 10-15 Pa during the freeze-drying process to obtain freeze-dried bacterial blocks. (4) Pulverization: The freeze-dried product from step (3) is pulverized and passed through an 80-mesh sieve to obtain freeze-dried powder; (5) Mixing and tableting: The lyophilized powder from step (4) is mixed with pharmaceutically acceptable excipients and tableted using a tablet press at a pressure of 20-25 kN to obtain the bacterial agent in the form of lyophilized tablets for vaginal use.

5. The preparation method according to claim 4, characterized in that: In step (2), the mass ratio of the freeze-drying protectant to Lactobacillus curvature cells is 2:1; Preferably, the freeze-drying protectant includes at least one of skim milk powder, trehalose, and sucrose; Preferably, the freeze-drying protectant contains 12 wt% skim milk powder, 8 wt% trehalose, and 5 wt% sucrose; Preferably, in step (5), the pharmaceutically acceptable excipients include at least one of fillers, disintegrants, and lubricants; Preferably, the vaginal lyophilized tablets comprise 30-35 parts of lyophilized powder, 25-30 parts of filler, 5-8 parts of disintegrant, and 1-2 parts of lubricant; Preferably, the filler is at least one of microcrystalline cellulose and lactose; Preferably, the disintegrant is at least one of crospovidone and low-substituted hydroxypropyl methylcellulose; Preferably, the lubricant is at least one of magnesium stearate and silicon dioxide; Preferably, each of the vaginal lyophilized tablets contains ≥1×10⁻⁶ live bacteria of Lactobacillus curvature OH126. 9 CFU; Preferably, the vaginal lyophilized tablets are packaged in aluminum-plastic blister packs containing an oxygen barrier layer, sealed and stored at 4°C, with a shelf life of up to 12 months.

6. The method for preparing the microbial agent according to claim 2 or 3, characterized in that: When the antifungal agent is in the form of a vaginal gel, the following steps are included: (1) Collection of bacterial cells: The Lactobacillus curvature OH126 described in claim 1 was inoculated into MRS liquid culture medium and cultured anaerobically at 37°C for 12-14 h until the late logarithmic growth phase. Then, it was centrifuged at 10000-12000 rpm for 15-20 min at 4°C. The supernatant was discarded and the live bacterial cells were collected. The entire process was carried out under aseptic conditions. (2) Preparation of bacterial suspension: Add sterile physiological saline to the bacterial cells in step (1) and adjust the concentration of the bacterial suspension to 1×10¹. 0 -2×10¹ 0 CFU / mL was used to obtain bacterial culture; (3) Gel matrix preparation: Take the gel matrix material and add it to purified water. Swell at room temperature for 24 hours, stirring 3-4 times during the period until a uniform and transparent gel matrix is ​​formed. Add humectant and preservative to the gel matrix, stir evenly, and then adjust the pH to 3.8-4.2 with pH adjuster. (4) Mixing: Under sterile conditions, slowly add the bacterial solution from step (2) to the gel matrix from step (3) while stirring at a rate of 50-60 rpm to ensure that the bacterial solution is evenly dispersed. Finally, add purified water to a total mass of 100g to form a vaginal gel. (5) Dispensing and sterilization: Dispense the vaginal gel prepared in step (4) into sterile pharmaceutical gel tubes and sterilize it by irradiation with 2-3 kGy dose of γ rays to obtain the bacterial agent in the form of vaginal gel.

7. The preparation method according to claim 6, characterized in that: In step (3), the gel matrix material includes at least one of carbomer 940 and hydroxypropyl methylcellulose; Preferably, the amount of the gel matrix material added is 1.5-2.0 g / 100 g gel; Preferably, the moisturizer includes at least one of glycerin and propylene glycol, and the amount of moisturizer added is 5-8g / 100g gel; Preferably, the preservative is phenoxyethanol, and the amount of preservative added is 0.3-0.5g / 100g gel; Preferably, the pH adjuster is at least one of citric acid and sodium citrate; Preferably, in step (5), each vaginal gel contains ≥1×10⁻⁶ live bacteria of Lactobacillus curvature OH126. 9 CFU; Preferably, the vaginal gel is sealed and stored at 4°C, with a shelf life of up to 6 months.

8. The use of the *Lactobacillus curvatureii* OH126 according to claim 1 or the bacterial agent according to claim 2 or 3 in the preparation of the product, characterized in that: The intended use is selected from any of the following: (1) Use in the preparation of products for the treatment or prevention of bacterial vaginosis; (2) Use in the preparation of products that help improve symptoms related to uterine fibroids, wherein the symptoms related to uterine fibroids include at least one of menstrual abnormalities and lower abdominal distension; (3) Use in the preparation of products that regulate the balance of the female vaginal microecology; (4) Use in the preparation of products that inhibit vaginal pathogens, wherein the vaginal pathogens include at least one of Gardnerella vaginalis, Prevotella vaginalis, Molybditis lappa and Candida albicans.

9. A product for treating or preventing bacterial vaginosis, characterized in that: The product contains Lactobacillus curvature OH126 as described in claim 1 or the bacterial agent as described in claim 2 or 3.

10. A product that helps improve symptoms related to uterine fibroids, characterized in that: The product contains Lactobacillus curvature OH126 as described in claim 1 or the bacterial agent as described in claim 2 or 3.

11. A product for regulating the vaginal microecological balance in women, characterized in that: The product contains Lactobacillus curvature OH126 as described in claim 1 or the bacterial agent as described in claim 2 or 3.

12. A product for inhibiting vaginal pathogens, characterized in that: The product contains Lactobacillus curvatureis OH126 as described in claim 1 or the bacterial agent as described in claim 2 or 3; the vaginal pathogens include at least one of Gardnerella vaginalis, Prevotella vaginalis, Molybditis lappa, and Candida albicans.

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