Method for improving vaginal environment of PCOS patient by oral administration of lactobacillus salivarius

Abstract

The invention discloses a method for improving the vaginal environment of a PCOS patient by orally taking lactobacillus salivarius. The method comprises the following steps: understanding and selecting oral lactobacillus salivarius suitable for the patient; the lactobacillus salivarius is correctly taken by making a taking plan; other auxiliary treatment means are combined for cooperative treatment; according to the method, the micro-ecological balance in the vagina of the PCOS patient is improved and maintained through metabolites secreted by the lactobacillus salivarius, and when the lactobacillus salivarius is selected, it is ensured that the oral lactobacillus salivarius does not contain components harmful to the body of the patient, so that the oral lactobacillus salivarius does not contain any component harmful to the body of the patient; the selected oral lactobacillus salivarius meets the use standard of PCOS patients, the existence form of the selected oral lactobacillus salivarius is ensured to be beneficial to better administration of the patients, and the selected lactobacillus salivarius is ensured to be reliable in source, high in activity, safe and free of side effects; and the lactobacillus salivarius can be effectively improved and maintained in the vagina environment of a PCOS patient in the follow-up process on the selection level of the lactobacillus salivarius.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbiological technology, in particular to a method for improving the vaginal environment of PCOS patients by oral administration of lactobacillus salivarius. BACKGROUND

[0002] PCOS patients, i.e. polycystic ovary syndrome patients, are endocrine syndromes characterized by long-term anovulation and high androgen, and are also the most common cause of menstrual disorders in women of childbearing age. The main clinical manifestations include menstrual abnormalities, ovulation disorders and infertility, and due to high androgen, can have hirsutism and acne. Lactobacillus salivarius belongs to the family of lactobacillaceae and the genus of lactobacillus, and is a kind of lactic acid bacteria with potential probiotic properties. It can produce secretory proteins that can destroy Staphylococcus aureus biofilm, thereby inhibiting Staphylococcus aureus. In addition, as a kind of edible lactic acid bacteria, lactobacillus salivarius can enhance immunity and regulate intestinal flora. PCOS patients often have changes in the vaginal environment, such as a decrease in the number of lactic acid bacteria and an increase in pH, which may increase the risk of infection. Oral administration of lactobacillus salivarius, a probiotic, can help improve the vaginal environment and reduce the risk of infection. Currently, when improving the condition of patients based on oral administration of lactobacillus salivarius, different patients have different reactions to lactobacillus salivarius. Therefore, it is not possible to formulate a targeted administration plan according to the individual constitution and condition of the patient when taking oral lactobacillus salivarius, which may lead to gastrointestinal discomfort in different patients after taking lactobacillus salivarius, thereby failing to improve the overall health status of the vaginal environment. SUMMARY

[0003] The present application provides a method for improving the vaginal environment of PCOS patients by oral administration of lactobacillus salivarius, which can effectively solve the problem of the current oral administration of lactobacillus salivarius for improving the condition of patients, as different patients have different reactions to lactobacillus salivarius. Therefore, it is not possible to formulate a targeted administration plan according to the individual constitution and condition of the patient when taking oral lactobacillus salivarius, which may lead to gastrointestinal discomfort in different patients after taking lactobacillus salivarius, thereby failing to improve the overall health status of the vaginal environment.

[0004] To achieve the above-mentioned purpose, the present application provides the following technical scheme: a method for improving the vaginal environment of PCOS patients by oral administration of lactobacillus salivarius, which regulates the balance of intestinal flora in patients by oral administration of lactobacillus salivarius, helps to improve the vaginal microecological environment of patients, thereby relieving related symptoms, and specifically includes the following steps: S1, understanding and selecting oral lactobacillus salivarius suitable for the patient; S2, correctly taking lactobacillus salivarius by formulating an administration plan; S3, combine other auxiliary treatment methods to cooperate with treatment; S4, according to the change of symptoms to carry on the corresponding monitoring and examination.

[0005] According to the above technical scheme, in S1, when selecting oral lactobacillus salivarius, it is necessary to understand that oral lactobacillus salivarius is a probiotic, and to select oral lactobacillus salivarius with high quality, high activity and high purity, while ensuring that the selected oral lactobacillus salivarius does not contain harmful ingredients to the patient's body, harmful ingredients including high sugar content and additives, and the selected oral lactobacillus salivarius needs to pay attention to the number of live bacteria, strain species and the use standard of oral lactobacillus salivarius for PCOS patients.

[0006] According to the above technical scheme, in S1, when selecting oral lactobacillus salivarius, it is necessary to understand that oral lactobacillus salivarius is a probiotic, and to select oral lactobacillus salivarius with high quality, high activity and high purity, while ensuring that the selected oral lactobacillus salivarius does not contain harmful ingredients to the patient's body, harmful ingredients including high sugar content and additives, and the selected oral lactobacillus salivarius needs to pay attention to the number of live bacteria, strain species and the use standard of oral lactobacillus salivarius for PCOS patients. At the same time, the selected oral lactobacillus salivarius of the appropriate form also needs to ensure that it comes from products produced by regular manufacturers and has been clinically verified to ensure that the selected oral lactobacillus salivarius is reliable, high in activity and safe without side effects.

[0007] According to the above technical scheme, in S2, during the preparation of the taking plan, the dose, the taking time and the periodic treatment course are determined; The determination of the dose is to determine the single oral lactobacillus salivarius dose according to the product instructions or the doctor's advice, and the oral lactobacillus salivarius dose can be adaptively changed according to the product activity, the patient's constitution and the patient's condition; The determination of the taking time refers to the determination of the taking time range of oral lactobacillus salivarius, which is generally within the time range of 30 minutes to 1 hour after meal to reduce the stimulation and discomfort of the stomach of the patient, increase the survival rate of lactobacillus salivarius in the intestinal tract, better play the role and ensure that the taking time is fixed within the specified time range after meal every day, help to maintain the stable level of probiotics in the patient's body; The periodic treatment course is determined according to the patient's individual condition and the doctor's advice to formulate a reasonable treatment course for the patient's condition. The specific condition of the patient is different, and the treatment course is also different. Usually, it needs to be treated for several weeks or months. During the treatment course of the patient, the treatment should not be interrupted, so as to improve and maintain the stable vaginal environment.

[0008] According to the above technical scheme, in S2, the taking plan is determined by determining the dose, the taking time and the periodic treatment course, and the oral lactobacillus salivarius is taken correctly according to the taking plan; Before the patient starts taking the oral Lactobacillus salivarius, a doctor needs to be consulted to determine whether the patient is suitable for taking probiotics, and the patient's medication being treated also needs to be determined to ensure that the patient's medication being treated does not interact with the oral Lactobacillus salivarius, and to avoid the patient's medication destroying the activity of Lactobacillus salivarius.

[0009] According to the above technical solution, in the S3, the specific auxiliary treatment means includes: balanced diet, moderate exercise, maintaining hygiene and reducing stress. The balanced diet refers to maintaining a balanced and healthy diet, increasing the intake of protein, vitamins and minerals, maintaining adequate water intake, reducing the intake of sugar and processed foods, improving the patient's immunity through dietary improvement, benefiting the diversification of the flora, helping the patient to improve the vaginal environment, and avoiding the intake of too much high-sugar and high-fat food to aggravate the patient's symptoms. The moderate exercise refers to promoting the patient's blood circulation, improving the patient's metabolism, and helping to improve the patient's overall health condition, and assisting in improving the patient's vaginal environment, and the selected exercise includes walking, jogging and yoga. The maintaining hygiene refers to keeping the patient's vagina clean by regularly washing with warm water, avoiding the use of strong cleaning products that can destroy the ecological balance of the patient's vaginal microorganisms. The reducing stress is to make the patient happy in body and mind by reducing stress, so as to assist the patient in regulating the endocrine system and further improving the symptoms of PCOS.

[0010] According to the above technical solution, in the S4, it is referred to as regularly conducting physical examination and monitoring related indicators during the patient taking Lactobacillus salivarius, so as to evaluate the treatment effect of Lactobacillus salivarius on the patient, and adjust the treatment scheme according to the result, which specifically includes observing the patient's symptom changes, regularly conducting gynecological examination and adjusting the patient's taking plan.

[0011] According to the above technical solution, the observation of the patient's symptom changes refers to regularly observing the changes of the patient's vaginal discharge, odor and itching symptoms, and timely communicating with the doctor, and in the case that the patient's symptoms are improved, the patient continues to take Lactobacillus salivarius according to the original taking plan. The regular gynecological examination refers to that after the patient takes Lactobacillus salivarius, the patient needs to regularly conduct gynecological examination within a specified time, and through the examination of related indicators, the changes of the patient's vaginal environment and the condition of PCOS are known. The adjustment of the patient's taking plan refers to evaluating the treatment effect of the patient according to the results of gynecological examination and symptom changes, and adjusting the treatment scheme of Lactobacillus salivarius together with the doctor according to the evaluation effect, so as to make the patient achieve the best treatment effect.

[0012] Compared with the prior art, the present application has the following beneficial effects: 1. By taking oral lactobacillus salivarius, the vaginal microecological balance of PCOS patients can be improved and maintained through the metabolic products secreted by lactobacillus salivarius, and in the process of selecting lactobacillus salivarius, it is ensured that the oral lactobacillus salivarius does not contain components harmful to the patient's body, so that the selected oral lactobacillus salivarius meets the use standard of PCOS patients, ensures that the existence form of the selected oral lactobacillus salivarius can benefit the patient to take better, and ensures that the selected lactobacillus salivarius is reliable in source, high in activity and safe without side effects, so as to ensure that it can effectively improve and maintain the vaginal environment of PCOS patients in the subsequent process.

[0013] 2. By formulating a taking plan for the patient, the dosage, taking time and treatment cycle of the patient taking lactobacillus salivarius can be determined, so that the patient's treatment process can be flexibly adapted according to the product activity, the patient's constitution and the patient's condition, and by ensuring that the patient's taking time every day is within the specified time range, the patient's body can be effectively helped to maintain a stable level of probiotics, reduce irritation and discomfort, and by formulating a targeted treatment course according to the specific condition of the patient, the patient can be effectively treated according to different conditions, so as to conveniently improve and maintain a stable vaginal environment more effectively.

[0014] 3. By combining the treatment scheme of oral lactobacillus salivarius with auxiliary treatment means, the patient's immunity can be enhanced by dietary improvement, the patient's intestinal flora can be diversified, the patient's vaginal environment can be improved, the patient's blood circulation can be promoted by exercise, the patient's overall health condition can be improved, the patient's vaginal environment can be improved, and by maintaining hygiene, avoiding hygiene problems from destroying the patient's vaginal microbial ecological balance, and adjusting the patient's stress to make the patient happy in body and mind, the patient's endocrine system can be adjusted and the PCOS symptoms can be improved, so that on the basis of oral lactobacillus salivarius treatment, the patient's vaginal environment can be more effectively improved by combining auxiliary treatment means.

[0015] 4. By regularly conducting physical examination and monitoring related indicators during the treatment of oral lactobacillus salivarius in patients, the improvement and treatment effect of lactobacillus salivarius on the patient's symptoms can be evaluated through the examination structure, the patient's treatment scheme can be adjusted according to the examination results, so that the patient can obtain the best treatment effect in the subsequent treatment process, and the patient's treatment condition can be detected and responded to in the patient's treatment process, so that corresponding adjustment can be made according to the patient's treatment condition in time, and the effect of improving the patient's symptoms can be further ensured. BRIEF DESCRIPTION OF DRAWINGS

[0016] The accompanying drawings are included to provide a further understanding of the application and are incorporated in and constitute a part of this specification, illustrate embodiments of the application and are intended to provide a further understanding of the application, and are made a part of the specification. Embodiments of the application will be described and explained with additional specificity and detail through the use of the accompanying drawings in which:

[0017] In the drawings: Figure 1 is a step flow chart of the improved method of the application. DETAILED DESCRIPTION

[0018] The preferred embodiments of the application will be described herein below with reference to the accompanying drawings, in which it is understood that the preferred embodiments described herein are illustrative only and are not intended to limit the present application.

[0019] Embodiment 1: As shown in the technical scheme provided by the present application, a method for improving the vaginal environment of a PCOS patient by orally taking Lactobacillus salivarius is provided, which adjusts the balance of the patient's intestinal flora by orally taking Lactobacillus salivarius, helps to improve the patient's vaginal microecological environment, and thus relieves related symptoms, and specifically includes the following steps: Figure 1 S1, understanding and selecting suitable oral Lactobacillus salivarius for the patient; S2, correctly taking Lactobacillus salivarius by formulating a taking plan; S3, combining other auxiliary treatment methods for treatment; S4, corresponding monitoring and examination according to the change of symptoms. Based on the above technical scheme, in S1, when selecting oral Lactobacillus salivarius, it is necessary to understand that oral Lactobacillus salivarius is a probiotic, and to select oral Lactobacillus salivarius containing high quality, high activity and high purity, while ensuring that the selected oral Lactobacillus salivarius does not contain harmful ingredients to the patient's body, the harmful ingredients specifically including high sugar content and additives, and the selected oral Lactobacillus salivarius needs to pay attention to the number of live bacteria, strain types of the product, and make the oral Lactobacillus salivarius meet the use standards of PCOS patients.

[0020] Based on the above technical scheme, in S1, when selecting oral Lactobacillus salivarius, it is necessary to understand that oral Lactobacillus salivarius is a probiotic, and to select oral Lactobacillus salivarius containing high quality, high activity and high purity, while ensuring that the selected oral Lactobacillus salivarius does not contain harmful ingredients to the patient's body, the harmful ingredients specifically including high sugar content and additives, and the selected oral Lactobacillus salivarius needs to pay attention to the number of live bacteria, strain types of the product, and make the oral Lactobacillus salivarius meet the use standards of PCOS patients.

[0021] At the same time, the selected oral Lactobacillus salivarius in the appropriate form also needs to ensure that it is a product produced by a regular manufacturer and has been clinically verified to ensure that the selected oral Lactobacillus salivarius is reliable in source, high in activity and safe without side effects.

[0022] ​Based on the above technical solution, in S2, the process of formulating the dosing plan specifically includes determining the dosage, determining the dosing time, and formulating the treatment cycle. The dosage is determined based on the product instructions or doctor's advice. The dosage of oral Lactobacillus salivarius can also be adjusted according to the product activity, the patient's constitution, and the patient's condition. Determining the timing of administration refers to determining the time range for oral administration of Lactobacillus salivarius. Generally, it should be taken within 30 minutes to 1 hour after a meal to reduce stomach irritation and discomfort, while increasing the survival rate of Lactobacillus salivarius in the intestines, allowing it to exert its effects better. It is also important to ensure that the daily administration time is fixed within the prescribed time range after meals to help maintain a stable level of probiotics in the patient's body. A treatment cycle is designed based on the patient's individual circumstances and the doctor's advice to create a reasonable treatment plan for the patient's condition. The duration of the treatment will vary depending on the patient's specific situation, and it usually requires treatment lasting for several weeks or months. During the treatment course, the patient should not interrupt the treatment in order to improve and maintain a stable vaginal environment.

[0023] Based on the above technical solution, in S2, the dosing plan is determined by determining the dosage, the dosing time, and the cycle course, and the Lactobacillus salivarius is taken correctly according to the dosing plan; Before a patient starts taking oral Lactobacillus salivarius, they need to consult a doctor to determine if the patient is suitable for taking probiotics. It is also necessary to determine the medications the patient is currently taking to ensure that they do not interact with the oral Lactobacillus salivarius and to avoid the medications destroying the activity of the Lactobacillus salivarius.

[0024] Based on the above technical solution, in S3, specific auxiliary treatment methods include: a balanced diet, moderate exercise, maintaining hygiene, and reducing stress; A balanced diet refers to maintaining a balanced and healthy diet, increasing the intake of protein, vitamins and minerals, maintaining adequate water intake, and reducing the intake of sugar and processed foods. Dietary improvements can help enhance the patient's immunity, promote vaginal flora diversity, help improve the vaginal environment, and avoid consuming too much high-sugar and high-fat food, which can aggravate the patient's symptoms. Moderate exercise refers to promoting blood circulation and increasing metabolism through moderate aerobic exercise, which helps improve the patient's overall health and assists in improving the vaginal environment. Suitable exercises include walking, jogging, and yoga. Maintaining hygiene means keeping the patient's vagina clean by regularly washing with warm water, avoiding the use of harsh cleaning products, and preventing disruption of the ecological balance of the patient's vaginal microbiota. Reducing stress is to make the patient happy in body and mind by reducing stress, so as to help the patient regulate the endocrine system and further improve the symptoms of PCOS.

[0025] Based on the above technical solution, S4 refers to regularly conducting physical examination and monitoring related indicators during the patient taking Lactobacillus salivarius, so as to evaluate the treatment effect of Lactobacillus salivarius on the patient, and adjust the treatment plan according to the results, which specifically includes observing the change of patient's symptoms, regularly conducting gynecological examination and adjusting patient's taking plan.

[0026] Based on the above technical solution, observing the change of patient's symptoms refers to paying attention to regularly observing the change of symptoms such as vaginal secretion, odor and itching of the patient, and communicating with the doctor in time. In the case of improvement of the patient's symptoms, continue to take Lactobacillus salivarius according to the original taking plan. Regularly conducting gynecological examination refers to that after taking Lactobacillus salivarius, the patient needs to regularly conduct gynecological examination within a specified time, and know the change of the vaginal environment and the condition of PCOS by checking related indicators; Adjusting the patient's taking plan refers to evaluating the treatment effect of the patient according to the results of gynecological examination and symptom change, and adjusting the treatment plan of Lactobacillus salivarius with the doctor according to the evaluation effect, so that the patient can achieve the best treatment effect.

[0027] Example 2: 1. Research object and sample collection: 1.1 Selection of research object 240 cases of 18-45 year-old patients with PCOS in the reproductive center of our hospital from January 2022 to June 2024 were selected as the research group. The diagnosis standard of PCOS refers to the 2003 Rotterdam standard. All research objects did not use hormone drugs within 3 months, and patients with endocrine diseases caused by other ovulation disorders and incomplete data or incomplete detection items were excluded. This study was approved by the medical ethics committee of our hospital (approval number IBR-20220015-R), and all patients signed the informed consent form.

[0028] 1.2 In vitro fertilization-embryo transfer clinical plan: Patients received gonadotropin-releasing hormone antagonist protocol, short long protocol in luteal phase, long protocol in follicular phase, prolonged protocol, minimal stimulation protocol or controlled ovarian hyperstimulation burst protocol treatment, when at least two follicles had a diameter > 18 mm, recombinant human chorionic gonadotropin (r-hCG) 10000 IU was used as a trigger for final oocyte maturation, oocyte retrieval was performed 36 h after r-hCG administration, day 3 blastulation stage embryo grading referred to the 2011 ESHRE Istanbul consensus, including cell number, fragmentation, symmetry, multinucleation, vacuoles and zona pellucida, at least one transferred embryo was a high quality embryo of grade 7A, 8A, 9A, 7B, 8B and 9B, ET was performed under transabdominal ultrasound guidance.

[0029] 1.3 Grouping and protocol PCOS patients 60 cases, divided into 2 groups, 30 cases in each group, namely PCOS group and PCOS + lactobacillus salivarius group, from the 0th day, PCOS + lactobacillus salivarius group takes a pack (oral) containing about 50 mg of freeze-dried probiotics (about 109 CFU of lactobacillus salivarius CECT5713, Baishi (Shanghai) Biotechnology Co., Ltd.) per day, for 3 months, after the diagnosis of pregnancy, oral administration of probiotics continues until the 15th week of pregnancy, during the entire study, the probiotics are stored at 4°C, and the patients are required to record the evaluation compliance in the form of clock-in, the low compliance rate (percentage of total treatment dose) is set to 86%, within 4 weeks before sampling, all participants are required to have not received hormone therapy, antibiotics or other probiotics, vaginal samples are collected at least 7 days after sexual intercourse to avoid or minimize the influence of partner semen on vaginal pH, microbial community composition or immune characteristics.

[0030] 1.4 Sample collection Patients were placed in lithotomy position, vaginal samples were collected from the upper third of the vagina with a sterile swab, the secretion on the cervical os was wiped off with sterile gauze, then the swab was rolled gently in the endocervical canal to collect the cervical sample, all sample swabs were placed in 2 mL Falcon tubes, immediately stored in liquid nitrogen for freezing, then transferred to the laboratory and stored at -80°C.

[0031] 1.5 DNA extraction The total genomic DNA of each sample was extracted using the QIAamp DNA Mini Kit, and the specific steps were as follows: before use, add anhydrous ethanol in buffer GD and rinse PW, add isopropanol in buffer GFA, add the volume reference bottle label, remove 200 μL of intestinal contents to a 2 mL centrifuge tube, and place the tube on ice, add 500 μL of buffer SA, 100 μL of buffer SC, and 15 μL of Proteinase K to the sample, mix using a TGrinder H24 tissue grinding homogenizer (OSE-TH-01) (oscillate at a speed of 6 M / S for 30 s, with an interval of 30 s, for a total of 2 cycles), incubate at 70 ℃ for 15 min, shake 2-3 times during incubation, vortex for 15 s, centrifuge at 12000 rpm (13400 g) for 3 min, transfer the supernatant to a new centrifuge tube, add 10 μL of RNase A, mix well after shaking, and let stand at room temperature for 5 min, add 200 μL of buffer SH, mix well, and place on ice for 5 min, centrifuge at 12000 rpm (13400 g) for 3 min, transfer the supernatant from the previous step to a new 1.5 mL centrifuge tube, add an equal volume of buffer GFA (add isopropanol), add the solution from the previous step to a CR2 adsorption column (place the adsorption column in a collection tube), centrifuge at 12000 rpm (13400 g) for 30 s, discard the waste, place the CR2 adsorption column in the collection tube, add 700 μL of rinse PW (check if anhydrous ethanol has been added before use) to the CR2 adsorption column, centrifuge at 12000 rpm (13400 g) for 30 s, discard the waste, and place the CR2 adsorption column in the collection tube, repeat the operation, place the CR2 adsorption column back into the collection tube, centrifuge at 12000 rpm (13400 g) for 2 min, discard the waste, and let the CR2 adsorption column stand at room temperature for 5 min to completely dry the residual rinse solution in the adsorption material, transfer the CR2 adsorption column to a clean centrifuge tube, add 50 μL of elution buffer TB to the middle of the adsorption membrane, let stand at room temperature for 2-5 min, centrifuge at 12000 rpm (13400 g) for 2 min, collect the solution in a centrifuge tube, measure the DNA concentration using a NanoDrop 2000, and store at -20 ℃ to prevent DNA degradation.

[0032] 2. The effect of oral Lactobacillus salivarius on the vaginal environment of PCOS patients 2.1 Vaginal pH determination and Nugent score The subjects were tested after 3 months of taking Salivaria lactis, the pH of the vaginal lateral wall and Nugent score, the pH test used Whatman pH paper (pH 3.8-5.5 and pH 6.0-8.1), the swab material was transferred to a glass slide, heated and fixed and subjected to Gram staining, the morphological type of Gram-positive, Gram-negative and Gram-variable bacteria was quantified, Nugent score 0-3 was considered normal, 4-6 was considered moderate, and 7-10 was considered bacterial vaginosis, the results showed that the vaginal pH determination and Nugent score of the PCOS+Salivaria lactis group were significantly lower than those of the PCOS group (Table 1) Table 1 Comparison of vaginal pH determination and Nugent score of the two groups

[0033] 2.2 Isolation and culture of lactic acid bacteria The other swab material was serially diluted and inoculated onto MRS agar plates supplemented with L-cysteine for the isolation of lactic acid bacteria, MRS medium was added with 0.5 g / L L-cysteine hydrochloride, adjusted to pH 6.2, autoclaved at 121 ℃ for 20 minutes, and after pouring the plates, an anaerobic environment was pre-equilibrated for 24 hours, the sample swab was vortexed in 1 mL of sterile saline, gradient diluted to 10⁻ 6 , 100 μL of each was spread on the plate and placed in an anaerobic workstation at 37 ℃ for 72 hours, different morphological colonies (diameter, color, edge characteristics) were selected and identified using a MALDI-TOF mass spectrometer (Bruker Biotyper), and a match score ≥ 2.0 was considered as the target strain.

[0034] 2.3 TaqMan qPCR detection of Salivaria lactis DNA DNA was extracted from each sample and the concentration was measured, specific primers and probes were used for fluorescent quantitative PCR detection, a 1:10 DNA dilution of Salivaria lactis CECT5713 (ranging from 2 ng to 0.2 pg) was used to calculate the concentration of the target bacterial genome, all samples and standards were repeated three times, the primer sequences were: forward primer For-Sal-3 (5'-GTCGTAACAAGGTAGCCGTAGGA-3') and reverse primer Rev-Sal-1 (5'-TAAACAAAGTATTCGATAAATGTACAGGTT-3'), probe SalivarISRDes-4 (5'-CGGCTGGATCACC-3').

[0035] Results showed that the relative abundance of lactic acid bacteria in PCOS+Lactobacillus salivarius group was significantly higher than that in PCOS group, and the DNA copy number of Lactobacillus salivarius in PCOS+Lactobacillus salivarius group was estimated to be (1.22±0.35)×10 7 Table 2. Comparison of relative abundance of lactic acid bacteria and DNA copy number of Lactobacillus salivarius between two groups

[0036] Table 2. Comparison of relative abundance of lactic acid bacteria and DNA copy number of Lactobacillus salivarius between two groups

[0037] 3. Effect of oral Lactobacillus salivarius on IVF-ET outcomes in PCOS patients 3.1 Comparison of the number of oocytes and the quality of oocytes between two groups of patients There was no significant difference in the total number of oocytes and mature oocytes between the PCOS group and the Lactobacillus salivarius group, but the maturation rate of oocytes in the PCOS group was slightly lower than that in the control group (Table 3).

[0038] Table 3. Comparison of the number of oocytes and the quality of oocytes between two groups of patients

[0039] 3.2 Comparison of embryo development indicators between two groups of patients The results showed that the number of mature oocytes, the number of oocyte fertilization, the number of normal oocyte fertilization, the number of blastocyst culture and the number of blastocyst formation in the PCOS group were significantly lower than those in the Lactobacillus salivarius group (P<0.001), as shown in Table 4.

[0040] Table 4. Comparison of embryo development indicators between two groups of patients

[0041] 3.3 Comparison of pregnancy indicators between two groups of patients Each patient was followed up, and the fetal heartbeat was detected by ultrasound 5 weeks after embryo transfer, which was considered as clinical pregnancy; the clinical pregnancy rate = the number of clinical pregnancy cycles / the number of transfer cycles×100%; the abortion rate = the number of abortion cycles / the number of clinical pregnancy cycles×100%; the live birth rate = the number of live births / the number of transfers×100%. The fresh cycle pregnancy outcomes of two groups of patients were analyzed, and the lost cycles were excluded. The results showed that the live birth rate in the Lactobacillus salivarius group was significantly higher than that in the PCOS group, and there was no significant difference in the clinical pregnancy rate and the abortion rate (P>0.05), as shown in Table 5.

[0042] Table 5. Comparison of pregnancy indicators between two groups of patients

[0043] Finally, it should be noted that the above only describes the preferred examples of the present application, and is not intended to limit the present application. Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art will appreciate that modifications can be made to the technical solutions described in the foregoing embodiments, or equivalent replacements can be made to some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for improving the vaginal environment in PCOS patients by oral administration of Lactobacillus salivarius, characterized in that: Oral administration of Lactobacillus salivarius can regulate the balance of the gut microbiota in patients, helping to improve the vaginal microecological environment and thus alleviate related symptoms. The specific steps include the following: S1. Understand and select the appropriate oral lactobacillus for the patient; S2. Properly take Lactobacillus salivarius by developing a dosage plan; S3. Combine with other auxiliary treatment methods to complement the treatment; S4. Conduct corresponding monitoring and examinations based on changes in symptoms.

2. The method for improving the vaginal environment of PCOS patients by oral administration of Lactobacillus salivarius according to claim 1, characterized in that: In S1, when selecting oral lactobacillus, it is necessary to understand that oral lactobacillus is a probiotic and to select oral lactobacillus containing high quality, high activity, and high purity. At the same time, it is necessary to ensure that the selected oral lactobacillus does not contain any components that are harmful to the patient's body. Harmful components specifically include high sugar content and additives. When selecting oral lactobacillus, attention should be paid to the number of live bacteria and the types of strains in the product, and the oral lactobacillus should meet the usage standards for PCOS patients.

3. The method for improving the vaginal environment of PCOS patients by oral administration of Lactobacillus salivarius according to claim 2, characterized in that: In S1, when selecting oral Lactobacillus salivarius, it is also necessary to determine the form in which the oral Lactobacillus salivarius product exists. The forms of oral Lactobacillus salivarius usually include capsules, granules, and chewable tablets. At the same time, the appropriate form of oral Lactobacillus salivarius should be selected, and it should be ensured that it comes from a reputable manufacturer and has undergone clinical validation.

4. The method for improving the vaginal environment of PCOS patients by oral administration of Lactobacillus salivarius according to claim 1, characterized in that: In S2, the process of developing a treatment plan specifically includes determining the dosage, determining the timing of administration, and developing a treatment cycle. The dosage is determined based on the product instructions or doctor's advice, and the dosage of oral Lactobacillus salivarius is also subject to adaptive changes based on product activity, patient's constitution and condition. The determination of the administration time refers to the determination of the time range for oral administration of Lactobacillus salivarius. It is generally taken within the range of 30 minutes to 1 hour after a meal to reduce stomach irritation and discomfort for patients, while increasing the survival rate of Lactobacillus salivarius in the intestines, so that it can play a better role, and to ensure that the administration time is fixed within the prescribed time range after meals every day. The aforementioned treatment cycle is designed based on the patient's individual circumstances and the doctor's advice to create a reasonable treatment plan for the patient's condition. The duration of the treatment will vary depending on the patient's specific situation, and it usually requires treatment lasting for several weeks or months. During the treatment course, patients should be careful not to interrupt the treatment.

5. A method for improving the vaginal environment of PCOS patients by oral administration of Lactobacillus salivarius according to claim 4, characterized in that: In S2, a dosing plan is determined by specifying the dosage, dosing time, and treatment cycle, and Lactobacillus salivarius is taken correctly according to the dosing plan; Before a patient starts taking oral Lactobacillus salivarius, they need to consult a doctor to determine if the patient is suitable for taking probiotics. It is also necessary to determine the medications the patient is currently taking to ensure that they do not interact with oral Lactobacillus salivarius.

6. A method for improving the vaginal environment of PCOS patients by oral administration of Lactobacillus salivarius according to claim 1, characterized in that: In S3, specific adjunctive treatment methods include: a balanced diet, moderate exercise, maintaining hygiene, and reducing stress; The so-called balanced diet refers to maintaining a balanced and healthy diet, increasing the intake of protein, vitamins and minerals, maintaining adequate water intake, reducing the intake of sugar and processed foods, and improving the patient's immunity through dietary improvements, which is conducive to the diversification of gut microbiota. The moderate exercise refers to promoting blood circulation, increasing metabolism, improving overall health, and assisting in improving the vaginal environment through moderate aerobic exercise. Selected exercises include walking, jogging, and yoga. Maintaining hygiene refers to keeping the patient's vagina clean by regularly washing with warm water, avoiding the use of harsh cleaning products, and preventing disruption of the ecological balance of the patient's vaginal microorganisms. The stress reduction mentioned above aims to alleviate stress and improve the patient's mental and physical well-being, thereby helping to regulate the endocrine system and further improve PCOS symptoms.

7. A method for improving the vaginal environment of PCOS patients by oral administration of Lactobacillus salivarius according to claim 1, characterized in that: S4 refers to conducting regular physical examinations and monitoring relevant indicators during the patient's use of Lactobacillus salivarius to assess the therapeutic effect of Lactobacillus salivarius on the patient and adjust the treatment plan based on the results. Specifically, this includes observing changes in the patient's symptoms, conducting regular gynecological examinations, and adjusting the patient's medication plan.

8. A method for improving the vaginal environment of PCOS patients by oral administration of Lactobacillus salivarius according to claim 7, characterized in that: The observation of changes in patient symptoms refers to regularly observing changes in vaginal discharge, odor, and itching, communicating with the doctor in a timely manner, and continuing to take Lactobacillus salivarius according to the original treatment plan if the patient's symptoms improve. The regular gynecological examination mentioned above refers to the requirement that patients undergo regular gynecological examinations within a specified time after taking Lactobacillus salivarius, in order to understand the changes in their vaginal environment and the condition of their PCOS by checking relevant indicators. Adjusting the patient's treatment plan refers to assessing the patient's treatment effectiveness based on gynecological examination results and changes in symptoms, and then working with the doctor to adjust the Lactobacillus salivarius treatment regimen based on the assessment results.

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