Biofilm Formation of Candida albicans Facilitates Fungal Infiltration and Persister Cell Formation in Vaginal Candidiasis.

BACKGROUND: Vaginal candidiasis is an important medical condition awaiting more effective treatment. How Candida albicans causes this disease and survives antifungal treatment is not yet fully understood. This study aimed to establish a comprehensive understanding of biofilm-related defensive strategies that C. albicans uses to establish vaginal candidiasis and to survive antifungal treatment. METHODS: A mouse model of vaginal candidiasis was adopted to examine the formation of biotic biofilms on the vaginal epithelium and fungal infiltration by laboratory and clinical strains of C. albicans. Histopathological changes and local inflammation in the vaginal epithelium caused by C. albicans of different biofilm phenotypes were compared. Antifungal susceptibility testing was carried out for C. albicans grown as planktonic cells, microplate-based abiotic biofilms, and epithelium-based biotic biofilms. Formation of persister cells by C. albicans in different growth modes was also quantified and compared. RESULTS: C. albicans wild-type reference strains and clinical isolates, but not the biofilm-defective mutants, formed a significant number of biotic biofilms on the vaginal epithelium of mice and infiltrated the epithelium. Biofilm formation and epithelial invasion induced local inflammatory responses and histopathological changes in the vaginal epithelium including neutrophil infiltration and subcorneal microabscesses. Biofilm growth on the vaginal epithelium also led to high resistance to antifungal treatments and promoted the formation of antifungal-tolerant persister cells. CONCLUSION: This study comprehensively assessed biofilm-related microbial strategies that C. albicans uses in vaginal candidiasis and provided experimental evidence to support the important role of biofilm formation in the histopathogenesis of vaginal candidiasis and the recalcitrance of the infection to antifungal treatment.

Using 17ß-estradiol heparin-poloxamer thermosensitive hydrogel to enhance the endometrial regeneration and functional recovery of intrauterine adhesions in a rat model.

Mechanical damage or infection to the endometrium can lead to the formation of adhesions in the uterine cavity, which may result in reduced reproductive outcome and/or pregnancy complications. The prognosis of this disease is poor due to few effective treatments and the complex environment of endometrium. Heparin-Poloxamer Hydrogel (HP hydrogel) is a nontoxic and biodegradable biomaterial, which has been commonly used as a sustained-release delivery system. In this study, we applied a mini-endometrial curette to scrape the endometrium of rats to mimic the process of curettage in patients. After the establishment of IUA model in rats, we injected the thermo-sensitive hydrogel(E2-HP hydrogel) into the injured uterine cavity and evaluated the therapeutic effect of E2-HP hydrogel on the recovery of IUA. Our results showed that E2-HP hydrogel can significantly facilitate the regeneration of injured endometrium along with inhibiting the cell apoptosis in IUA model. Furthermore, we revealed that E2-HP hydrogel on the recovery of IUA was closely associated with the upregulation of kisspeptin through activating the ERK1/2 and MAPKs p38 pathways. In conclusion, E2-HP hydrogel can effectively transfer E2 into the injured endometrium and it can be considered as a promising therapeutic method for the women with intrauterine adhesions.

SNHG5 enhances Paclitaxel sensitivity of ovarian cancer cells through sponging miR-23a.

BACKGROUND: Ovarian cancer is one of the most lethal gynecological malignancies throughout the world. Long non-coding RNA (lncRNA) small nucleolar RNA host gene 5 (SNHG5) has been reported to play an important role in several human cancers, but the role of SNHG5 in the chemoresistance of ovarian cancer cells is yet elusive. METHOD: The expression of SNHG5 and miR-23a were determined by quantitative reverse transcriptase polymerase chain reaction. The effects of SNHG5 and miR-23a on the sensitivity of ovarian cancer cells to paclitaxel (PTX) were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and flow cytometry. The subcellular location of SNHG5 was detected by a subcellular fraction assay. The interaction between SNHG5 and miR-23a was determined by luciferase reporter assay and RNA immunoprecipitation assay. RESULTS: The expression of SNHG5 was downregulated in the cancer genome atlas cohort. Similarly, decreased expression of SNHG5 was observed in ovarian cancer tissues. Moreover, lower expression of SNHG5 was found in PTX-resistant ovarian cancer patients as well as PTX-resistant ovarian cancer cell lines. Downregulation of SNHG5 expression was indicative of poor prognosis in patients with ovarian cancer. Overexpression of SNHG5 enhanced the sensitivity of SKOV3/PTX and HeyA-8/PTX cells to PTX in vitro and enhanced PTX sensitivity in tumors in vivo. Interestingly, an inverse correlation between SNHG5 and miR-23a expression was found in ovarian cancer tissues and SNHG5 functioned as a decoy for miR-23a. Silencing of miR-23a overcame the resistance of SKOV3/PTX and HeyA-8/PTX cells to PTX. More importantly, miR-23a overexpression could reverse the inductive effect of SNHG5 overexpression on PTX sensitivity of ovarian cancer cells. CONCLUSION: SNHG5 enhanced the sensitivity of ovarian cancer cells to PTX through sponging miR-23a, providing a new mechanism of chemoresistance in ovarian cancer.

RAFT-Derived Polymethacrylates as a Superior Treatment for Recurrent Vulvovaginal Candidiasis by Targeting Biotic Biofilms and Persister Cells.

BACKGROUND: Vulvovaginal candidiasis (VVC) is a common infection in need of more effective treatment. Formation of epithelium-associated Candida biofilms and the presence of persister cells are among the major contributing factors to the recurrence of this condition. We have previously developed RAFT-derived polymethacrylates that are effective in killing C. albicans biofilms in vitro. This study aimed to examine the clinical potential of polymethacrylates as antifungals for treatment of recurrent VVC (RVVC). METHODS: A mouse model of VVC was used to establish vaginal epithelium-associated biofilms, using C. albicans isolates from VVC/RVVC patients. A comparison was made of the efficacies of polymethacrylates and conventional antifungals, clotrimazole and nystatin, in killing Candida in epithelium-associated biofilms in vivo. Ex vivo biofilms were used for Candida population profiling and to quantify persister cells in vaginal epithelia. The potency of polymethacrylates and conventional antifungals against persister cells, either as sole agents or in combination, was assessed. RESULTS: Polymethacrylates showed negligible local toxicity, resistance to vaginal acidity, and outstanding in vivo activity against vaginal epithelium-associated C. albicans biofilms. In vivo tests polymethacrylates outperformed the conventional antifungals, nystatin and clotrimazole at concentrations 50 times below the over-the-counter concentrations. Using polymethacrylates was associated with fewer persister cells, and better eradication of persister cells pre-selected by conventional antifungals. CONCLUSION: This study systematically assessed the clinical potential of RAFT-derived polymethacrylates as an effective treatment for VVC/RVVC in a mouse model. Polymethacrylates effectively killed vaginal epithelium-related C. albicans in vivo by specially targeting biotic biofilms and persister cells. Treatment presented negligible local toxicity.

Analyzing the clinical significance of postoperative methotrexate in the management of early abdominal pregnancy: analysis of 10 cases.

OBJECTIVES: To assess the clinical value and treatment outcomes of postoperative methotrexate (MTX) therapy in themanagement of early abdominal pregnancy. MATERIAL AND METHODS: We retrospectively analyzed ten (10) cases of early abdominal pregnancy at our hospital between7th August, 2006 and 20th April, 2017. RESULTS: Out of the ten (10) cases identified, six (6) patients and four (4) patients underwent surgery (laparotomy or laparoscopy)only and surgery (laparotomy or laparoscopy) plus IM 50 mg/m2 methotrexate (MTX) within 24 hours of surgeryrespectively. The gestation age and serum ß-HcG levels were significantly lower (p < 0.05, 6.0 ± 1.82 and 8073.2 ± 9561.0)in the surgery plus MTX group in comparison to (7.33 ± 3.61 and 15625 ± 21275.2) for the surgery only group. Ultrasoundimaging findings reported extra uterine pregnancy in all cases and diagnostic surgery was necessary to locate precise siteof implantation to plan further treatment. Days of hospitalization were shorter in the surgery + MTX group than in thesurgery only group (3.00 ± 0.816 versus 5.66 ± 2.80). CONCLUSIONS: Earliness in diagnosis coupled with the appropriate (methotrexate) MTX regime could help prevent unwantedcomplications that could arise from delayed or misdiagnosis.

Metformin Promotes Regeneration of the Injured Endometrium Via Inhibition of Endoplasmic Reticulum Stress-Induced Apoptosis.

Intrauterine adhesion (IUA) is now recognized as one of the most common diseases in reproductive-age women. Metformin, a well-known frontline oral antidiabetic drug, has been found effective in numerous different diseases. The aim of this study was to determine the effect of metformin on reducing adhesions in an animal model of IUA. Sprague-Dawley rats were randomized into 4 groups: sham operation, control, metformin-treated for 7 days, and metformin-treated for 14 days. To establish the IUA model, mechanical injury to the endometria of rats was induced with a mini curette. Metformin was injected intraperitoneally after surgery. A significant amelioration in both the number of glands and the fibrotic area, compared to those of the control group, was detected 14 days after metformin intervention. The expression levels of antigen KI-67 and vascular endothelial growth factor were increased at 7 and 14 days after treatment. However, the transforming growth factor-ß expression was decreased at 14 days after treatment. Endoplasmic reticulum stress-related apoptosis proteins (glucose-regulated protein 78, caspase-12, and CCAAT/enhancer binding protein (EBP) homologous protein) were downregulated after metformin treatment. Moreover, we determined that the effect of metformin was related to the inhibition of endoplasmic reticulum stress-induced apoptosis via the Phosphatidylinositol 3 kinase (PI3K)/Protein kinase B (AKT) and Extracellular regulated protein kinases1/2 pathways. In conclusion, metformin can attenuate the adhesion and promote the regeneration of the endometrium of the IUA rat, and metformin may serve as a novel therapeutic strategy for IUA patients.