The goal of this clinical trial is to explore the effect of Lactobacillus johnsonii and white pomegranate extract on female bacterial status, it aims to answer are:

• Evaluate Lactobacillus johnsonii TCI250 probiotics or white pomegranate extract in regulating female vaginal bacteria and improving vaginal health.
• Evaluate Lactobacillus johnsonii TCI250 probiotics or white pomegranate extract in regulating inflammation.

Participants will be randomly assigned to placebo (n = 50), probiotic heat-killed Lactobacillus johnsonii TCI250 group (n = 50) and white pomegranate extract group (n = 50) for 8 weeks. Researchers will compare the vaginal flora and inflammation.

This study is an interventional trial conducted in a randomized double-blind controlled study, we will recruit 150 participants. Participants will be randomly d divided into three groups (Group A: Placebo; Group B: heat-killed Lactobacillus johnsonii TCI250; Group C: White pomegranate extract, dosage of 500mg) for 8 weeks. Subsequently, participants will be followed up for an additional 8 weeks to observe changes in the vaginal microbiota and inflammation. At weeks 0, 4, 8, and 16, participants will record basic information such as personal dietary habits, lifestyle, and health conditions. Additionally, vaginal epithelial cells and whole blood (5 mL) will be collected at weeks 0, 8, and 16 for liver and kidney function monitoring (GPT, creatinine).

1. Study Participants i. Inclusion Criteria (1) Females aged 20-45 (2) Nugent score ≥ 4 (3) Menstrual cycle of 28-60 days ii. Exclusion Criteria (1) Pregnant or lactating women (2) Antibiotic usage (3) Other reproductive-related inflammatory diseases (e.g., candidiasis, gonorrhea, chlamydia infection) (4) Allergy to probiotics or White Pomegranate-related components

   iii. Withdrawal Criteria Participants are free to decide whether to participate in this trial, and they can withdraw their consent at any time during the trial without providing any reason. If participants experience discomfort, they may withdraw from the trial at any time without facing any penalties or compromising their rights.

2. Subject Recruitment Interested participants will be recruited openly through posters posted at Chung Shan Medical University Hospital. The researchers will explain the trial content for 15-20 minutes, and if interested, participants will sign the informed consent form.

3. Research Methods i. Questionnaire assessment ii. Nugent score evaluation iii. Inflammatory markers detection: Including IL-4, IL-10, IL-6, IL-8, MCP-1, TNF-α, IL-1β. I iv. Microbiota gene sequencing: Utilizing 16S rRNA gene sequences to analyze the vaginal microbiota.

4. Statistical Methods Data from this study will be analyzed using SigmaPlot statistical software (version 12.5; Systat Software, San Jose, CA) and SAS statistical software (version 9.4, Statistical Analysis System Institute Inc., Cary, NC, USA). One-way ANOVA will be used to compare differences among the three groups, repeated measurement ANOVA to compare pre- and post-intervention values within each group, and multiple linear regression to analyze the impact of supplement intake on vaginitis scores, inflammation, and microbiota diversity. The results will be presented as mean ± standard deviation (SD). Statistical significance will be indicated by p < 0.05.

Through this study, the administration of Lactobacillus johnsonii TCI250 probiotics and White Pomegranate extract is investigated for their effectiveness in improving the vaginal microbiota in women. This could potentially serve as a means of promoting vaginal health and well-being.

Inclusion Criteria:

1. Females aged 20-45
2. Nugent score ≥ 4
3. Menstrual cycle of 28-60 days

Exclusion Criteria:

1. Pregnant or lactating women
2. Antibiotic usage
3. Other reproductive-related inflammatory diseases (e.g., candidiasis, gonorrhea, chlamydia infection)
4. Allergy to probiotics or White Pomegranate-related components