Quercetin antagonized advanced glycated end products induced apoptosis and functional inhibition of fibroblasts from the prolapsed uterosacral ligament.

The altered behaviors and functions of pelvic floor fibroblasts are pathophysiological changes of pelvic organ prolapse (POP). Our previous study showed that advanced glycated end products (AGEs) accumulated in the pelvic tissues of POP and induced fibroblast apoptosis. The study was designed to investigate whether quercetin antagonize AGEs-induced apoptosis and functional inhibition of fibroblasts. The uptake of 5-ethynyl-2'-deoxyuridine (EdU) was evaluated for cell proliferation. Flow cytometric analysis was applied for cell apoptosis. Intracellular reactive oxygen species (ROS) content was determined by the fluorescence of dichlorofluorescein (DCF). The contractility of fibroblasts was measured by collagen gel contraction assay. The expressions of extracellular matrix (ECM) related genes and the expression of miR-4429 and caspase-3 were quantified by qPCR. The expressions of phosphatase and tensin homolog (PTEN), phosphoinositide 3-kinase (PI3K), serine-threonine kinase (Akt), and phosphorylated Akt (p-Akt) were analyzed by Western Blot. The down-regulation of miR-4429 was achieved by cell transfection. Quercetin antagonized AGEs-induced apoptosis, proliferation inhibition, and ROS increase in fibroblasts. Quercetin did not alleviate AGEs-induced contractile impairment of fibroblasts. Quercetin reduced the gene expressions of lysyl oxidase like protein 1 (LOXL1)and matrix metallopeptidase 1 (MMP1), and increased the gene expressions of lysyl oxidase (LOX) and fibrillin 2 (FBN2) in fibroblasts. Quercetin reversed AGEs-induced upregulation of PTEN and downregulation of PI3K, P-Akt, and miR-4429 in fibroblasts. The inhibitory effect of quercetin on AGEs-induced fibroblast apoptosis was inhibited by downregulating the expression of miR-4429. In conclusion, quercetin antagonized AGEs-induced apoptosis and functional inhibition of fibroblasts from the prolapsed uterosacral ligament. And inhibiting AGEs-induced down-regulation of miR-4429/PTEN/PI3K/Akt pathway was the mechanism underlying the antagonistic effect of quercetin on AGEs-induced apoptosis.

Exosomes derived from mesenchymal stromal cells: a promising treatment for pelvic floor dysfunction.

Pelvic floor dysfunction (PFDs), which include pelvic organ prolapse (POP), stress urinary incontinence (SUI) and anal incontinence (AI), are common degenerative diseases in women that have dramatic effects on quality of life. The pathology of PFDs is based on impaired pelvic connective tissue supportive strength due to an imbalance in extracellular matrix (ECM) metabolism, the loss of a variety of cell types, such as fibroblasts, muscle cells, peripheral nerve cells, and oxidative stress and inflammation in the pelvic environment. Fortunately, exosomes, which are one of the major secretions of mesenchymal stromal cells (MSCs), are involved in intercellular communication and the modulation of molecular activities in recipient cells via their contents, which are bioactive proteins and genetic factors such as mRNAs and miRNAs. These components modify fibroblast activation and secretion, facilitate ECM modelling, and promote cell proliferation to enhance pelvic tissue regeneration. In this review, we focus on the molecular mechanisms and future directions of exosomes derived from MSCs that are of great value in the treatment of PFD.

CD106/VCAM-1 distinguishes a fibroblast subpopulation with high colony-forming capacity and distinct protein expression from the uterosacral ligament.

Background: Pelvic organ prolapse (POP) is a common degenerative disease in women which may diminish quality of life. Investigating the pathological changes of the uterosacral ligament, including the functional changes of fibroblasts, is critical to understanding the pathophysiology of POP. This study was designed to isolate CD106-positive (CD106+) fibroblasts from the human uterosacral ligament and assess the function and expression of this subpopulation. Methods: We separated CD106+ fibroblasts and CD106 negative (CD106-) fibroblasts by fluorescence-activated cell sorting (FACS) and cultured them for subsequent experiments. Flow cytometric analysis was used to test the sorting efficiency, CD106 expression, and typical mesenchymal stem cell (MSC) phenotype marker expression. A colony-forming unit (CFU) assay was applied to evaluate the colony-forming ability of the fibroblasts. Trilineage differentiation capacities were assessed after in vitro induction. The protein levels of vimentin, fibroblast specific protein-1 (FSP-1), collagen I (COL 1), matrix metallopeptidase-1 (MMP-1), and α-smooth muscle actin (α-SMA) were detected by western blot analysis. The expression of CD106 was verified by flow cytometric analysis and immunohistochemistry (IHC) in the POP and non-POP groups. Results: The CD106+ fibroblasts were isolated with a purity of (93.50±3.91)%. The CD106+ fibroblasts exhibited higher colony-forming capacity than that of CD106- fibroblasts, but neither of them showed adipogenic or osteogenic differentiation similar to that of MSCs. The protein levels of MMP-1 and α-SMA were lower, and the level of COL 1 was higher in the CD106+ fibroblasts than in the CD106- fibroblasts. In addition, we observed a decreased expression of CD106 in the POP group compared with the non-POP group. Conclusions: Our results suggest that CD106+ fibroblasts possess a high colony-forming capacity and distinct protein expression, and this subpopulation is reduced in POP.

Advanced glycation end products (AGEs) downregulate the miR-4429/PTEN axis to promote apoptosis of fibroblasts in pelvic organ prolapse.

Background: Pelvic organ prolapse (POP) is a common degenerative disease among females. We previously reported that advanced glycation end products (AGEs), compounds derived from nonenzymatic glycoxidation reactions, accumulated in the human vaginal wall and impaired the function of fibroblasts in the pathogenesis of POP. This study investigated the apoptosis induced by AGEs in human uterosacral ligament fibroblasts and the underlying mechanism. Methods: Human uterosacral ligament fibroblasts were cultured and identified. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was performed to identify the expression of miR-4429, phosphatase and tensin homolog (PTEN), and caspase-3. Flow cytometric analysis was applied to detect the apoptosis rate of fibroblasts. Dual-luciferase reporter assay was performed to verify the relationship between miR-4429 and PTEN. The overexpression of miR-4429 and the inhibition of PTEN were achieved by cell transfections. Western blot analysis was used to detect the protein levels of PTEN, phosphoinositide 3-kinase (PI3K), and protein kinase B (Akt). Results: The AGEs promoted fibroblast apoptosis both in the POP and the non-POP groups. The expression of PTEN increased in fibroblasts from the POP group or fibroblasts treated with AGEs. It was confirmed that miR-4429 interacted with PTEN messenger RNA (mRNA), and the expression level of miR-4429 was reduced in fibroblasts from the POP group or fibroblasts treated with AGEs. Further, overexpression of miR-4429 alleviated increased PTEN expression and fibroblast apoptosis induced by AGEs. Similarly, inhibition of PTEN expression alleviated increased fibroblast apoptosis induced by AGEs. In addition, the protein expressions of PI3K and phosphorylated Akt were reduced in fibroblasts exposed to AGEs. Conclusions: We proposed that AGEs induced fibroblast apoptosis by regulating the miR-4429/PTEN/PI3K/Akt pathway in POP. Our results revealed a novel mechanism by which AGEs contributed to the molecular pathological alteration in POP.

Transvaginal single-port versus multi-port laparoscopic sacrocolpopexy: a retrospective cohort study.

BACKGROUND: Sacrocolpopexy is the gold standard treatment for apical prolapse. With the development of minimally invasive surgical techniques, the new approach of transvaginal single-port laparoscopic sacrocolpopexy (TS-LSC) has become available. However, its therapeutic effects remain unclear. The aim of this study is to compare the middle-term clinical outcomes of transvaginal single-port laparoscopic sacrocolpopexy with multi-port laparoscopic sacrocolpopexy (LSC) for apical prolapse. METHODS: We conducted a retrospective cohort study. Patients with advanced apical prolapse who underwent either TS-LSC or LSC between May 2017 to June 2019 were enrolled. Baseline demographics, perioperative results, perioperative and postoperative complications, pelvic organ prolapse quantification (POPQ) scores, pelvic floor distress inventory (PFDI-20) score and pelvic organ prolapse/urinary incontinence sexual function questionnaire (PISQ-12) score were collected at 2 years. RESULTS: 89 subjects were analyzed: 46 in TS-LSC and 43 in LSC group. Follow-up time was 38.67 ± 7.46 vs 41.81 ± 7.13 months, respectively. Baseline characteristics and perioperative outcomes were similar except that pain score was lower (2.37 ± 0.90 vs 3.74 ± 1.05) and cosmetic score was higher (9.02 ± 0.75 vs 7.21 ± 0.89) in TS-LSC group (P < 0.05). Complication rates did not differ between groups. 3 mesh exposure in each group were noted. Recurrence rate was 2.17% in TS-LSC and 6.98% in LSC, no apical recurrence occurred. Constipation was the most common postoperative symptom. Besides, patients in TS-LSC group had better POP-Q C point (- 6.83 ± 0.54 vs - 6.39 ± 0.62, P < 0.05), and similar Aa, Ap and TVL values. Bladder and pelvic symptoms were improved in both groups, but colorectal symptoms were not relieved. There were no differences of PISQ-12 scores between groups. CONCLUSION: TS-LSC was not inferior to LSC at 2 years. Patients may benefit from its mild pain, better cosmetic effect and better apical support as well as good safety and efficacy. TS-LSC is a promising considerable choice for advanced vaginal apical prolapse. Trial registration ChiCTR2000032334, 2020-4-26 (retrospectively registered).

The Clinical Value and Variation of Antithyroid Antibodies during Pregnancy.

Antithyroid antibodies, which include thyroid-stimulating hormone receptor antibodies (TRAbs), thyroid peroxidase antibodies (TPOAbs), and thyroid globulin antibodies (TgAbs), are widely known for their tight association with thyroid autoimmune diseases. The variation in all three kinds of antibodies also showed different trends during and after pregnancy (Weetman, 2010). This article reviewed the the physiological changes, while focusing on the variation of thyroid antibodies concentration in women during and after pregnancy, and adverse consequences related to their elevation. Since abnormal elevations of these antithyroid antibodies may lead to adverse outcomes in both mothers and fetuses, special attention must be paid to the titer of the antibodies during pregnancy. The molecular mechanisms of the variations in those antibodies have yet to be explained. The frequency and timing of thyroid antibody measurement, as well as different reference levels, also remain to be elucidated.

MSC-based therapy in female pelvic floor disorders.

Mesenchymal stem cells (MSCs), also referred to as multipotent stromal cells or mesenchymal stromal cells, are present in multiple tissues and capable of differentiating into diverse cell lineages, holding a great promise in developing cell-based therapy for a wide range of conditions. Pelvic floor disorders (PFDs) is a common degenerative disease in women and may diminish a woman's quality of life at any age. Since the treatments for this disease are limited by the high rates of recurrence and surgical complications, seeking an ideal therapy in the restoration of pelvic floor function is an urgent issue at present. Herein, we summarize the cell sources of MSCs used for PFDs and discuss the potential mechanisms of MSCs in treating PFDs. Specifically, we also provide a comprehensive review of current preclinical and clinical trials dedicated to investigating MSC-based therapy for PFDs. The novel therapy has presented promising therapeutic effects which include relieving the symptoms of urinary or fecal incontinence, improving the biological properties of implanted meshes and promoting the injured tissue repair. Nevertheless, MSC-based therapies for PFDs are still experimental and the unstated issues on their safety and efficacy should be carefully addressed before their clinical applications.

The cytotoxicity of advanced glycation end products was attenuated by UCMSCs in human vaginal wall fibroblasts by inhibition of an inflammatory response and activation of PI3K/AKT/PTEN.

Pelvic organ prolapse (POP) occurs when the pelvic organs (bladder, bowel or uterus) herniate into the vagina, causing incontinence, voiding, and bowel and sexual dysfunction, negatively impacting upon a woman's quality of life. Intermediate intermolecular cross-links and advanced glycation cross-links increase in prolapsed tissue. Stem cells are able to participate in tissue repair due to their ability to differentiate into multiple lineages, and thus into various types of connective tissue cells, so they therefore hold great promise for treating pelvic floor dysfunction. The current study found that advanced glycation end products (AGEs) inhibited the viability and proliferation of human vaginal wall fibroblasts (VWFs), were cytotoxic to VWFs, and also induced the apoptosis of VWFs. In contrast, umbilical cord-derived mesenchymal stem cells (UCMSCs) secreted anti-inflammation cytokines to protect against the cytotoxic effects of fibroblasts induced by AGEs and attenuated the cytotoxic effect of AGE on fibroblasts by activation of the PI3K/Akt-PTEN pathway. This study demonstrated that UCMSCs inhibited the cytotoxic effect of AGE in cells from patients with POP by inducing an anti-inflammatory reaction and activating the PI3K/AKT/PTEN signaling pathway. The current results provide important insights into use of stem cells to treat POP.

New thoughts in exploring the pathogenesis, diagnosis, and treatment of threatened abortion.

Threatened abortion is a common complication of pregnancy. Since the underlying mechanisms behind this condition are complicated, predicting and treating threatened abortion is a challenge for clinicians. Interestingly, a recent article in Bioscience Trends (Biosci Trends 2019; DOI: 10.5582/bst.2019.01111) revealed a higher, not lower, level of ꞵ-human chorionic gonadotropin (hCG) and estrogen during the first 6 weeks of pregnancy, suggesting a novel association between ꞵ-hCG, estrogen, and threatened abortion. Unfortunately, this study was limited by its small sample size, unconvincing trial design, and inadequate exploration of the underlying mechanisms. This low-quality evidence indicates that a higher level of ꞵ- hCG and estrogen is associated with threatened abortion. However, that work provided some new insights for further studies of threatened abortion.

Sperm DNA fragmentation valued by SCSA and its correlation with conventional sperm parameters in male partner of recurrent spontaneous abortion couple.

The objective of this study is to evaluate the predictive value of sperm DNA fragmentation Index (DFI) in unexplained recurrent spontaneous abortion (RSA) and to investigate its correlation with conventional sperm parameters. Besides, we aimed to reveal the necessity of establishing a DFI clinical threshold of each laboratory for the prognostic diagnosis of RSA and establish our own DFI threshold. Semen samples were collected from male partners of RSA patients (n = 139) and healthy recent fathers (control, n = 200). DFI was tested using SCSA and conventional semen analysis was performed using an automatic semen analyzer. The DFI value and distribution were compared between the two groups using corresponding statistical software. The diagnostic threshold value was established by ROC curve. The correlation between DFI and the conventional semen parameters of the 139 cases was further analyzed using Student's t test and Mann-Whitney U test. Our result showed that DFI was significantly higher in RSA patients compared with normal donor controls. We established our own DFI threshold at 13.59%. There was only a weak partial correlation between DFI values and conventional sperm analysis parameters. Our present study suggested that DFI might be used as a valuable predictor for RSA independent of conventional sperm parameters. Additionally, we recommend that each laboratory should establish its own clinical DFI threshold for more precise prediction of RSA and we recommend that sperm DNA fragmentation test should be included in complete sperm quality assessment in addition to conventional semen analysis for RSA male partners.

Parkinson's disease-like motor and non-motor symptoms in rotenone-treated zebrafish.

The pesticide rotenone is widely used to produce Parkinson's disease (PD)-like symptoms in rodents, but few studies have examined whether rotenone-treated zebrafish can serve as an animal model of PD. Here, we report that 4 weeks of rotenone treatment induced motor and non-motor PD-like symptoms in adult zebrafish. Compared with control fish, rotenone-treated fish spent less time swimming at a fast speed, indicating a deficit in motor function. In the light-dark box test, rotenone-treated fish exhibited longer latencies to enter the dark compartment and spent more time in the light compartment, reflecting anxiety- and depression-like behavior. Furthermore, rotenone-treated fish showed less of an olfactory preference for amino acid, indicating olfactory dysfunction. These behavioral symptoms were associated with decreased levels of dopamine in the brains of rotenone-treated fish. Taken together, these results suggest that rotenone-treated zebrafish are a suitable model of PD.