This study aimed to explore the mechanism of n-butanol alcohol extract of Baitouweng Decoction(BAEB) in the treatment of vulvovaginal candidiasis(VVC) in mice based on the negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis. In the experiment, female C57BL/6 mice were divided randomly into the following six groups: a blank control group, a VVC model group, high-, medium-, and low-dose BAEB groups(80, 40, and 20 mg·kg~(-1)), and a fluconazole group(20 mg·kg~(-1)). The VVC model was induced in mice except for those in the blank control group by the estrogen dependence method. After modeling, no treatment was carried out in the blank control group. The mice in the high-, medium-, and low-dose BAEB groups were treated with BAEB at 80, 40, and 20 mg·kg~(-1), respectively, and those in the fluconazole group were treated with fluconazole at 20 mg·kg~(-1). The mice in the VVC model group received the same volume of normal saline. The general state and body weight of mice in each group were observed every day, and the morphological changes of Candida albicans in the vaginal lavage of mice were examined by Gram staining. The fungal load in the vaginal lavage of mice was detected by microdilution assay. After the mice were killed, the degree of neutrophil infiltration in the vaginal lavage was detected by Papanicolaou staining. The content of inflammatory cytokines interleukin(IL)-1β, IL-18, and lactate dehydrogenase(LDH) in the vaginal lavage was tested by enzyme-linked immunosorbent assay(ELISA), and vaginal histopathology was analyzed by hematoxylin-eosin(HE) staining. The expression and distribution of NLRP3, PKCδ, pNLRC4, and IL-1Ra in vaginal tissues were measured by immunohistochemistry(IHC), and the expression and distribution of pNLRC4 and IL-1Ra in vaginal tissues were detected by immunofluorescence(IF). The protein expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by Western blot(WB), and the mRNA expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by qRT-PCR. The results showed that compared with the blank control group, the VVC model group showed redness, edema, and white secretions in the vagina. Compared with the VVC model group, the BAEB groups showed improved general state of VVC mice. As revealed by Gram staining, Papanicolaou staining, microdilution assay, and HE staining, compared with the blank control group, the VVC model group showed a large number of hyphae, neutrophils infiltration, and increased fungal load in the vaginal lavage, destroyed vaginal mucosa, and infiltration of a large number of inflammatory cells. BAEB could reduce the transformation of C. albicans from yeast to hyphae. High-dose BAEB could significantly reduce neutrophil infiltration and fungal load. Low-and medium-dose BAEB could reduce the da-mage to the vaginal tissue, while high-dose BAEB could restore the damaged vaginal tissues to normal levels. ELISA results showed that the content of inflammatory cytokines IL-1β, IL-18, and LDH in the VVC model group significantly increased compared with that in the blank control group, and the content of IL-1β, IL-18 and LDH in the medium-and high-dose BAEB groups was significantly reduced compared with that in the VVC model group. WB and qRT-PCR results showed that compared with the blank control group, the VVC model group showed reduced protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues of mice and increased protein and mRNA expression of NLRP3. Compared with the VVC model group, the medium-and high-dose BAEB groups showed up-regulated protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues and inhibited protein and mRNA expression of NLRP3 in vaginal tissues. This study indicated that the therapeutic effect of BAEB on VVC mice was presumably related to the negative regulation of NLRP3 inflammasome by promoting PKCδ/NLRC4/IL-1Ra axis.
Female ; Animals ; Humans ; Mice ; Candidiasis, Vulvovaginal/drug therapy* ; Inflammasomes/genetics* ; Interleukin-18 ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; 1-Butanol/pharmacology* ; Fluconazole/therapeutic use* ; Interleukin 1 Receptor Antagonist Protein/therapeutic use* ; Mice, Inbred C57BL ; Candida albicans ; Cytokines ; Drugs, Chinese Herbal/pharmacology* ; Ethanol ; RNA, Messenger ; Calcium-Binding Proteins/therapeutic use*

This study aims to explore the effect of butyl alcohol extract of Baitouweng Decoction(BAEB) on vulvovaginal candidiasis(VVC) in mice and to clarify the mechanism from Toll-like receptors(TLRs)/MyD88 and Dectin-1/Syk signal pathways and NLRP3 inflammasome. To be specific, female KM mice were randomized into control group(i.g., normal saline), model group, fluco-nazole group(i.g., 20 mg·kg~(-1)), and low-dose, medium-dose, and high-dose BAEB groups(i.g., 20, 40, and 80 mg·kg~(-1), respectively). VVC was induced in mice except the control group. After the modeling, administration began and lasted 7 days. The ge-neral conditions and body weight of mice were recorded every day. On the 1 st, 3 rd, 7 th, and 14 th after vaginal infection by Candida albicans, the fungal load in the vaginal lavage fluid of the mice was measured with the plate method, and the morphology of C. albicans in vaginal lavage fluid was observed based on Gram staining. After the mice were killed, vaginal tissues were subjected to hematoxylin-eosin(HE) staining and periodic acid-Schiff(PAS) staining for vaginal histopathological analysis. The content of cytokines in vaginal lavage fluid, such as interleukin(IL)-1β, IL-18, tumor necrosis factor-α(TNF-α), IL-6, and S100 a8, was determined by enzyme-linked immunosorbent assay(ELISA), and content of reactive oxygen species(ROS) in vaginal tissues by tissue ROS detection kit. The protein expression of NLRP3, ASC, caspase-1, Dectin-1, Syk, MyD88, TLR2, TLR4, and nuclear factor-κB(NF-κB) in vaginal tissues was detected by Western blot, and the levels and distribution of NLRP3, Dectin-1, Syk, MyD88, TLR2, and TLR4 in vaginal tissues were determined with the immunohistochemical method. The results show that BAEB can improve the general conditions of VVC mice, reduce the fungal load and C. albicans hyphae in vaginal secretion, decrease ROS content in vaginal tissues and content of cytokines in vaginal lavage fluid, and down-regulate the expression of NLRP3, ASC, caspase-1, Dectin-1, Syk, MyD88, TLR2, TLR4, and NF-κB in vaginal tissues. The above results indicate that BAEB exerts therapeutic effect on VVC mice by down-regulating the key proteins in the TLRs/MyD88 and Dectin-1/Syk signal pathways and NLRP3 inflammasome.
1-Butanol/therapeutic use* ; Animals ; Candida albicans ; Candidiasis, Vulvovaginal/drug therapy* ; Caspase 1/metabolism* ; Cytokines/metabolism* ; Female ; Humans ; Inflammasomes/metabolism* ; Mice ; Myeloid Differentiation Factor 88/metabolism* ; NF-kappa B/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Plant Extracts/therapeutic use* ; Reactive Oxygen Species/metabolism* ; Signal Transduction ; Toll-Like Receptor 2/metabolism* ; Toll-Like Receptor 4/metabolism* ; Tumor Necrosis Factor-alpha/metabolism*

Background: Vulvovaginal Candidiasis (VVC) is one of the frequent infections of the female genital tract and is the second most common cause of vaginal infections after bacterial vaginosis. According to the Centers for Disease Control and Prevention, azoles are the first‑line treatment for VVC. Among the azoles available in the Philippines, only miconazole and clotrimazole are recommended for both pregnant and non‑pregnant women. Objective: Compare the effect of miconazole versus clotrimazole in the treatment of vulvovaginal candidiasis among patients seen at the out‑patient department in a tertiary hospital Materials and Methods: This involved review of the records of patients diagnosed with VVC in a tertiary medical center from 2016 to 2020. All records of women, pregnant and non‑pregnant, wherein single‑dose 1200 mg miconazole or 6‑day 100 mg clotrimazole given vaginally were included Results: Eleven out of the 316 records (3.46%) remained symptomatic after treatment, about 18.1% (2/161) from those who used miconazole and 81.8% (9/155) from those treated with clotrimazole (p 0.027). In terms of failure rate, for miconazole it was 1.2% (2/161), whereas for clotrimazole it was 5.8% (9/155). None of the charts were found to have recorded adverse reaction to the given treatment Conclusion Single‑dose miconazole intravaginal regimen has a higher clinical cure rate than the 6‑day clotrimazole intravaginal treatment. Thereby, single‑dose intravaginal miconazole has the potential to improve patient compliance and treatment outcome at a lower cost
Clotrimazole ; Miconazole ; Vaginitis ; Candidiasis, Vulvovaginal

OBJECTIVES: Human papillomavirus (HPV) is a kind of spherical DNA virus, which is related to many factors such as immune status and pregnancy. Due to the decrease of immunity, pregnant women are more likely to have HPV infection, which causes serious imbalance of vaginal microecology and is not beneficial to pregnancy outcome. Therefore, this study focuses on the impact of HPV infection on vaginal microecology and maternal and neonatal outcomes. METHODS: A total of 140 pregnant women with HPV infection during pregnancy, who received obstetric examination in the First Affiliated Hospital of Hainan Medical College from November 2017 to July 2019, were selected as a HPV infection group, and 150 normal pregnant women with HPV negative in the same period were selected as a control group. Vaginal secretions were collected from all the pregnant women at 28-34 weeks of gestation to evaluate vaginal pH, cleanliness and microecological status, and to record pregnancy outcomes for all pregnant women. RESULTS: The proportions of vaginal pH>4.5, constituent ratio of flora density and diversity of I-II, positive detection rate of vulvovaginal candidiasis (VVC) and bacterial vaginosis (BV) in HPV infected pregnant women were significantly higher than those in the control group (all CONCLUSIONS Pregnant women with HPV infection during pregnancy are more likely to have vaginal microecological disorders, and can increase the risk of adverse pregnancy outcomes such as premature delivery and chorioamnionitis.
Candidiasis, Vulvovaginal ; Cesarean Section ; Female ; Humans ; Infant, Newborn ; Papillomavirus Infections/epidemiology* ; Pregnancy ; Vaginosis, Bacterial

To investigate the effects of butyl alcohol extract of Baitouweng Decoction(BAEB) on neutrophil chemotaxis in vaginal mucosa of mice with vulvovaginal candidiasis(VVC). Seventy-two SPF female Kunming mice were randomly divided into normal control group, model group, fluconazole group, BAEB low-dose group, middle-dose group and high-dose group. Subcutaneous injection of estradiol benzoate was conducted to induce pseudo-estrus, and then 2×10~6 CFU·mL~(-1)of Candida albicans was inoculated into vaginal lumen, followed by drug treatment for 7 days. Gram staining was used to observe the morphological changes of C. albicans in vagina; vaginal fungal load was detected on agar plate. Histological changes of vaginal tissues in mice were observed by HE staining. Lactate dehydrogenase(LDH), interleukin-6(IL-6) and tumor necrosis factor(TNF-α) levels in mouse lavage fluid were detected by enzyme-linked immunosorbent assay(ELISA). Neutrophils in vaginal lavage fluid was observed and counted by using Pap smear. The levels of IL-8 and MIP-2 in vaginal mucosa were detected by ELISA. IL-8 and MIP-2 mRNA levels in vaginal mucosa of mice were detected by qRT-PCR. The results showed that as compared with the normal group, VVC model group had a large number of hyphae and a high level of fungal loadinvagina. The vaginal mucosa was completely destroyed, the number of neutrophils increased, and the protein and mRNA levels of IL-8 and MIP-2 were up-regulated. After BAEB treatment, the hyphae of the treatment group was decreased, the fungal load was decreased, the impaired mucosa showed different degrees of improvement, the inflammatory factors were decreased to varying degrees, and the protein and mRNA levels of chemokine IL-8 and MIP-2 were down-regulated. In conclusion, BAEB may be used to treat VVC by inhibiting vulvovaginal candidiasis via blocking neutrophils recruitment into vagina.
1-Butanol ; Animals ; Candida albicans ; Candidiasis, Vulvovaginal/drug therapy* ; Chemotaxis/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Female ; Mice ; Mucous Membrane/drug effects* ; Neutrophils/drug effects* ; Vagina/diagnostic imaging*

Candidiasis, Vulvovaginal ; microbiology ; Communicable Diseases ; microbiology ; Female ; Humans ; Microbiota ; physiology ; Vagina ; microbiology ; Vaginitis ; microbiology ; Vaginosis, Bacterial ; microbiology

OBJECTIVE: To evaluate the efficacy of cis-2-dodecenoic acid (BDSF) in the treatment and prevention of vaginal candidiasis in vivo. METHODS: The activities of different concentrations of BDSF against the virulence factors of Candida albicans (C. albicans) were determined in vitro. An experimental mouse model of Candida vaginitis was treated with 250 μmol/L BDSF. Treatment efficiency was evaluated in accordance with vaginal fungal burden and inflammation symptoms. RESULTS: In vitro experiments indicated that BDSF attenuated the adhesion and damage of C. albicans to epithelial cells by decreasing phospholipase secretion and blocking filament formation. Treatment with 30 μmol/L BDSF reduced the adhesion and damage of C. albicans to epithelial cells by 36.9% and 42.3%, respectively. Treatment with 200 μmol/L BDSF completely inhibited phospholipase activity. In vivo mouse experiments demonstrated that BDSF could effectively eliminate vaginal infection and relieve inflammatory symptoms. Four days of treatment with 250 μmol/L BDSF reduced vaginal fungal loads by 6-fold and depressed inflammation. Moreover, BDSF treatment decreased the expression levels of the inflammatory chemokine-associated genes MCP-1 and IGFBP3 by 2.5- and 2-fold, respectively. CONCLUSION BDSF is a novel alternative drug that can efficiently control vaginal candidiasis by inhibiting the virulence factors of C. albicans.
Animals ; Candida albicans ; drug effects ; metabolism ; pathogenicity ; physiology ; Candidiasis, Vulvovaginal ; drug therapy ; genetics ; immunology ; microbiology ; Chemokine CCL2 ; genetics ; immunology ; Disease Models, Animal ; Fatty Acids, Monounsaturated ; administration & dosage ; Female ; Fungal Proteins ; genetics ; metabolism ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; genetics ; immunology ; Mice ; Virulence ; drug effects ; Virulence Factors ; genetics ; metabolism

Bacterial vaginosis (BV) and complicated vulvovaginal candidiasis (VVC) are frequently occurring vaginal infections in postmenopausal women, caused by an imbalance in vaginal microflora. Postmenopausal women suffer from decreased ovarian hormones estrogen and progesterone. A normal, healthy vaginal microflora mainly comprises Lactobacillus species (spp.), which act beneficially as a bacterial barrier in the vagina, interfering with uropathogens. During premenopausal period, estrogen promotes vaginal colonization by lactobacilli that metabolizing glycogen and producing lactic acid, and maintains intravaginal health by lowering the intravaginal pH level. A lower vaginal pH inhibits uropathogen growth, preventing vaginal infections. Decreased estrogen secretion in postmenopausal women depletes lactobacilli and increases intravaginal pH, resulting in increased vaginal colonization by harmful microorganisms (e.g., Enterobacter, Escherichia coli, Candida, and Gardnerella). Probiotics positively effects on vaginal microflora composition by promoting the proliferation of beneficial microorganisms, alters the intravaginal microbiota composition, prevents vaginal infections in postmenopausal. Probiotics also reduce the symptoms of vaginal infections (e.g., vaginal discharge, odor, etc.), and are thus helpful for the treatment and prevention of BV and VVC. In this review article, we provide information on the intravaginal mechanism of postmenopausal vaginal infections, and describes the effectiveness of probiotics in the treatment and prevention of BV and VVC.
Candida ; Candidiasis, Vulvovaginal ; Colon ; Enterobacter ; Escherichia coli ; Estrogens ; Female ; Glycogen ; Humans ; Hydrogen-Ion Concentration ; Lactic Acid ; Lactobacillus ; Microbiota ; Odors ; Postmenopause ; Premenopause ; Probiotics* ; Progesterone ; Vagina ; Vaginal Discharge ; Vaginal Diseases ; Vaginosis, Bacterial

OBJECTIVETo investigate the vaginal flora in patients with recurrent vulvovaginal candidiasis (RVVC).

METHODSVaginal swabs were collected at different time points from 6 RVVC patients and 5 healthy women of child-bearing age. The dynamic changes, microbiota composition, alpha diversity and beta diversity in the two groups were assessed by analyzing the 16S rRNA V4 hypervariable region amplified from the total genomic DNA from the swabs.

RESULTSLactobacillus was the predominant species in healthy women with similar proportions of L.iners and L.crispatus; small proportions of Gardnerella, Prevotella and other genus were also detected. In some healthy women, the vaginal flora showed a high relative abundance of anaerobic bacteria such as Gardnerella, Prevotella, Atopobium, Sneathia. Compared with the healthy women, patients with RVVC showed a significantly reduced diversity of vaginal flora, where L.iners was the predominant species and the content of L.crispatus decreased significantly. In healthy women, the vaginal flora fluctuated with the menstrual cycle, and the fluctuation was the most prominent during menstruation; the dominant species either alternated regularly or maintain an absolute superiority in the menstrual cycle. The vaginal flora showed attenuated fluctuation in women with RVVC, were highly conserved within the menstrual cycle, and maintained a similar composition in the episodes and intermittent periods.

CONCLUSIONThe vaginal flora of RVVC patients do not undergo regular variations with the menstrual cycle and shows a similar composition between the episodes and intermittent periods. Promoting the production of L.iners or inhibiting the colonization of L.crispatus to restore the composition of the vaginal flora may help in the treatment of RVVC.

Candidiasis, Vulvovaginal ; microbiology ; Case-Control Studies ; Female ; Humans ; Lactobacillus ; classification ; isolation & purification ; Longitudinal Studies ; Menstrual Cycle ; Microbiota ; RNA, Ribosomal, 16S ; isolation & purification ; Vagina ; microbiology

BACKGROUNDVulvovaginal candidiasis (VVC) was a common infection associated with lifelong harassment of woman's social and sexual life. The purpose of this study was to describe the species distribution and in vitroCandidaCandida spp.) isolated from patients with VVC over 8 years.

METHODSSpecies which isolated from patients with VVC in Peking University First Hospital were identified using chromogenic culture media. Susceptibility to common antifungal agents was determined using agar diffusion method based on CLSI M44-A2 document. SPSS software (version 14.0, Inc., Chicago, IL, USA) was used for statistical analysis, involving statistical description and Chi-square test.

RESULTSThe most common strains were Candida (C.) albicans, 80.5% (n = 1775) followed by C. glabrata, 18.1% (n = 400). Nystatin exhibited excellent activity against all species (<4% resistant [R]). Resistance to azole drugs varied among different species. C. albicans: clotrimazole (3.1% R) < fluconazole (16.6% R) < itraconazole (51.5% R) < miconazole (54.0% R); C. glabrata: miconazole (25.6% R) < clotrimazole (50.5% R) < itraconazole (61.9% R) < fluconazole (73.3% R); Candida krusei: clotrimazole (0 R) < fluconazole (57.7% R) < miconazole (73.1% R) < itraconazole (83.3% R). The susceptibility of fluconazole was noticeably decreasing among all species in the study period.

CONCLUSIONSNystatin was the optimal choice for the treatment of VVC at present. The species distribution and in vitroCandida spp. isolated from patients with VVC had changed over time.

Antifungal Agents ; pharmacology ; Candida ; drug effects ; pathogenicity ; Candidiasis, Vulvovaginal ; microbiology ; China ; Clotrimazole ; pharmacology ; Drug Resistance, Fungal ; Female ; Fluconazole ; pharmacology ; Humans ; Itraconazole ; pharmacology ; Miconazole ; pharmacology ; Microbial Sensitivity Tests