Characterization of chitosan-gelatin cryogel templates developed by chemical crosslinking and oxidation resistance of camellia oil cryogel-templated oleogels.

In this study, chitosan-gelatin conjugates were prepared by chemical crosslinking of tannic acid. The cryogel templates were developed through freeze-drying and immersed in camellia oil to construct cryogel-templated oleogels. Chemical crosslinking resulted in apparent colour changes and improved emulsion-related/rheological properties on conjugates. The cryogel templates with different formulas exhibited different microstructures with high porosities (over 96 %), and crosslinked samples might have higher hydrogen bonding strength. Tannic acid crosslinking also led to enhanced thermal stabilities and mechanical properties. Cryogel templates could reach a considerable oil absorption capacity of up to 29.26 g/g and prevent oil from leaking effectively. The obtained oleogels with high tannic acid content possessed outstanding antioxidant abilities. After 8 days of rapid oxidation at 40 °C, Oleogels with a high degree of crosslinking owned the lowest POV and TBARS values (39.74 nmol/kg, and 24.40 µg/g, respectively). This study indicates that the involvement of chemical crosslinking would favor the preparation and the application potential of cryogel-templated oleogels, and the tannic acid in the composite biopolymer systems could act as both the crosslinking agent and the antioxidant.

Polymerizable Surfactant Ligand for Stabilization and Film Formation of CsPbBr3 Nanocrystals.

Surfactant ligands are important in the synthesis of inorganic perovskite nanocrystals (NCs), not only for stabilizing NCs but also for surface defect passivation. A new polymerizable surfactant ligand with a multidentate l-cysteine head, a long oleoyl tail, and a polymerizable styrenyl group (NOSVC) is designed for the post-synthesis treatment and stabilization of colloidal CsPbBr3 NCs in this work. 1H nuclear magnetic resonance and X-ray photoelectron spectroscopy analysis show that the l-cysteine head has strong interactions with the NCs. The absolute photoluminescence quantum yields of the colloidal NCs are increased from 45.1% of the pristine NCs stabilized with oleic acid/oleyl amine to 91.8% after NOSVC treatment. NOSVC-stabilized CsPbBr3 colloidal NCs show enhanced stabilities when exposed in polar solvents. The NOSVC-stabilized CsPbBr3 NCs in a solid film state allow for a photopolymerization to be carried out with the assistance of a photoinitiator. The polymerized films of NOSVC-treated NCs exhibit significantly enhanced stability against thermal radiation, ultraviolet irradiation, and humidity. We also fabricated self-healing polymer films incorporating NOSVC-treated CsPbBr3 NCs as a green filter for a white light-emitting diode device. The green light-emitting films are very stable in humid environments, revealing the great application potential of NOSVC-treated CsPbBr3 NCs in flexible display and lighting devices.

Effects of different concentrations of sodium hypochlorite on dentine adhesion and the recovery application of sodium erythorbate. / ä¸åŒæµ“åº¦æ¬¡æ°¯é ¸é’ å¤„ç†å¯¹ç‰™æœ¬è´¨ç²˜æŽ¥çš„å½±å“åŠå¼‚æŠ—åè¡€é ¸é’ çš„æ¢å¤ä½œç”¨.

OBJECTIVES: Root canal therapy is the most effective and common method for pulpitis and periapical periodontitis. During the root canal preparation, chemical irrigation plays a key role. However, sodium hypochlorite (NaOCl), the widely used irrigation fluid, may impact the bonding strength between dentin and restorative material meanwhile sterilization and dissolving. Therefore, it's important to explore the influence of NaOCl on the adhesion between dentin and restoration materials to ensure clinical efficacy. This study aims to explore the effect of NaOCl on dentine adhesion and evaluate the effect of dentine adhesion induced by sodium erythorbate (ERY), and to provide clinical guidance on dentin bonding after root canal therapy. METHODS: Seventy freshly complete extracted human third molars aged 18-33 years old, without caries and restorations were selected. A diamond saw was used under running water to achieve dentine fragments which were divided into 10 groups with 14 fragments in each group: 2 control [deionized water (DW)±10% ERY] and 8 experimental groups (0.5%, 1%, 2.5%, and 5.25% NaOCl±10% ERY). The dentine specimens in the control group (treated with DW) and the experimental groups (treated with 0.5% NaOCl, 1% NaOCl, 2.5% NaOCl, and 5.25% NaOCl) were immersed for 20 min using corresponding solutions which were renewed every 5 min. The other 5 groups were immersed in 10% ERY for 5 min after an initial washing with DW for 1 min. Then, we selected 4 dentine fragments from all 14 fragments in each group and the numbers and diameters of opening dentinal tubules were observed under scanning electron microscope (SEM). The other 10 dentine fragments from each group were used to make adhesive samples by using self-etch adhesive wand composite resin. All the above adhesive samples were sectioned perpendicular to the bonded interface into 20 slabs with a cross-sectional area of 1 mm×1 mm using a diamond saw under the cooling water, and then the morphology of 10 slabs in each group's bonding interface was observed from aspects of formation of resin tags, depth of tags in dentin, and formation of hybrid layer under SEM. The other 10 slabs of each group's microtensile bond strength and failure modes were also analyzed. RESULTS: Among the 0.5% NaOCl, 1% NaOCl, 2.5% NaOCl, and 5.25% NaOCl groups, the number and diameter of patent dentinal tubules gradually increased with the rise of concentration of NaOCl solution (all P<0.05). Among the DW, 0.5% NaOCl, 1% NaOCl, 2.5% NaOCl, and 5.25% NaOCl groups, the number and diameter of patent dentinal tubules increased after using ERY, but without significant difference (all P>0.05). Among the DW, 0.5% NaOCl, 1% NaOCl, and 2.5% NaOCl groups, the scores of formation of resin tags under SEM gradually increased with the increase of concentration of NaOCl solution, while the score in the 5.25% NaOCl group decreased significantly compared with the score of the 2.5% NaOCl group (P<0.05). There was no significant difference between using 10% ERY groups and without using 10% ERY groups (all P>0.05). The scores of length of the tags under SEM in the 5.25% NaOCl group was significantly higher than the scores of DW, 0.5% NaOCl, and 1% NaOCl groups (all P<0.05), and it was also higher than the score of the 2.5% NaOCl group, but without significant difference (P>0.05). There was no significant difference between using 10% ERY groups and without using 10% ERY groups (P>0.05). The scores of formation of hybrid layer under SEM in the 2.5% NaOCl and 5.25% NaOCl groups significantly decreased compared with the score of the DW group (all P<0.05). There were significant differences between the 2.5% NaOCl±10% ERY groups and between the 5.25% NaOCl±10% ERY groups (all P<0.05). Microtensile bond strength was greater in the 0.5% NaOCl, 1% NaOCl, and 2.5% NaOCl groups, but lower in the 5.25% NaOCl group than that in the DW group (all P<0.05). There were significant differences between the 2.5% NaOCl±10% ERY groups and between the 5.25% NaOCl±10% ERY groups (all P<0.05). The incidence of type "Adhesive" of failure modes in the 5.25% NaOCl group was significantly higher than that in other groups (all P<0.05), while the incidence of type "Adhesive" in the 5.25% NaOCl+10% ERY group was lower than that in the 5.25% NaOCl group (P<0.05). CONCLUSIONS: The bonding strength to dentine increases with the increase of NaOCl concentration when the concentration lower than 2.5%; whereas it is decreased at a higher concentration (such as 5.25%). 10% ERY has a definite recovery effect on attenuated bonding strength to 5.25% NaOCl-treated dentine.

A two-year water-stable 2D MOF with aqueous NIR photothermal conversion ability.

Structural lability in humid air or water severely limits the practical use of MOFs. Developing new MOFs with exceptional water stability is interesting for both industrial applications and academic research. Herein we report a new method to improve the water stability of MOFs by using three-dimensional rigid shielding ligands. A very highly stable two-dimensional MOF (CuCP-MOF) is synthesized in this work, in which [2,2]paracyclophane dicarboxylate ligands are coordinated with Cu(ii) ions to form a paddle wheel structure. CuCP-MOF is a triclinic crystal with unit cell parameters a = 10.065 Å, b = 10.897 Å, c = 10.940 Å, α = 90.676°, ß = 91.729°, and γ = 92.725° determined by single crystal X-ray diffraction and DFT simulation. It can easily form MOF nanosheets due to the large interlayer distance and weak interlayer interactions. It shows good aqueous stability, and remains intact after storage in water for two years, as evidenced by FTIR and XRD analyses. CuCP-MOF shows a strong absorption in the NIR range due to the d-d transition of Cu(ii). The aqueous dispersions of CuCP-MOF exhibit high NIR photothermal conversion efficiency, about 17.5% for a laser with an energy density of 5 W cm-2 (808 nm) and 22.0% for a laser of 2 W cm-2 on average.

The exosomal integrin α5ß1/AEP complex derived from epithelial ovarian cancer cells promotes peritoneal metastasis through regulating mesothelial cell proliferation and migration.

PURPOSE: Epithelial ovarian cancer (EOC) is one of the most malignant cancers in the gynecologic system. Many patients are diagnosed at an advanced stage with disseminated intra-peritoneal metastases. EOC spreads via both direct extension and trans-coelomic spread. However, the interplay between human peritoneal mesothelial cells (HPMCs) and EOC cells is still ambiguous. We hypothesize that integrins (ITG) in HPMCs may play important roles in EOC metastasis. METHODS: The expression of different integrin subtypes from HPMCs was assessed using Western blotting. The expression of integrin α5ß1 (ITGA5B1) and its co-localization with asparaginyl endopeptidase (AEP) in HPMCs derived from EOC patients (EOC-HPMCs) were assessed using immunofluorescence. The role and mechanism of the exosomal ITGA5B1/AEP complex in HPMCs was assessed using both in vitro and in vivo assays. A retrospective study involving 234 cases was carried out to assess ITGA5B1 and AEP levels in circulating sera and ascites of EOC patients, as well as associations between ITGA5B1/AEP expression and overall survival. RESULTS: We found that ITGA5B1was highly expressed and co-localized with AEP in EOC cells, and that the exosomal ITGA5B1/AEP complex secreted by EOC cells played an important role in the proliferation and migration of HPMCs. High levels of exosomal ITGA5B1/AEP were also found in circulating sera and ascites of EOC patients, and the expression of ITGA5B1/AEP in EOC tissues was found to be negatively associated with overall survival. CONCLUSIONS: Our data indicate that EOCs may regulate the function of HPMCs through exosomal ITGA5B1/AEP, which may be crucial for peritoneal metastasis.

Development and internal validation of a Nomogram for preoperative prediction of surgical treatment effect on cesarean section diverticulum.

BACKGROUND: The aim of this study was to develop and validate an individualized score based on preoperative parameters to predict patient outcomes after vaginal repair of cesarean section diverticulum. METHODS: This is a retrospective cohort study (Canadian Task Force classification II-2). Patients were enrolled between Jun 11, 2012, to May 27, 2016. Multivariable logistic regression analyses were used to construct the predictive model. Then, we generated a nomogram to assess the individualized risk of poor prognosis after operation. This prediction model included information from 167 eligible patients diagnosed with cesarean section diverticulum who underwent vaginal repair. Class-A healing group was defined as CSD patients who had menstruation duration of no more than 7 days and a thickness of the remaining muscular layer of no less than 5.8 mm after vaginal repair according to conferences. Others were included in the non-class-A healing group. A final nomogram was computed using a multivariable logistic regression model. RESULTS: The factors contained in the individualized prediction nomogram included the depth/ the thickness of the remaining muscular layer ratio, number of menstruation days before surgery, White blood cell and fibrinogen. This model demonstrated adequate discrimination and calibration (C-index = 0.718). There was a significant difference in the number of postmenstrual spotting days (12.98 ± 3.86 VS 14.46 ± 2.86, P = 0.022) and depth/ the thickness of the remaining muscular layer ratio (2.81 ± 1.54 VS 4.00 ± 3.09, P = 0.001) between two groups. Decision curve analysis showed that this nomogram was clinically useful. CONCLUSIONS: This cesarean section diverticulum score can predict the outcomes of cesarean section diverticulum and can be useful for counseling patients who are making treatment decisions.

MUC16 impacts tumor proliferation and migration through cytoplasmic translocation of P120-catenin in epithelial ovarian cancer cells: an original research.

BACKGROUND: Epithelial ovarian cancer (EOC) remains one of the most lethal gynecologic cancers, and its pathogenetic mechanism remains unclear. Here we show that MUC16 promotes the translocation of p120-catenin (p120ctn) to the cytoplasm and consequently activates ras homolog (Rho) GTPases RhoA/Cdc42 activation to modulate the proliferation and migration abilities of EOC cells. METHODS: We collect 94 ovarian cancer (OC) patients' tissue samples to constitute tissue microarray (TMA) and analyze the MUC16 and p120ctn expression levels. Lentivirus transfection is used to overexpress cytoplasmic tail domain (CTD) of MUC16 and CRISPR/Cas9 genome-editing system is firstly used to knock out MUC16 in EOC cells. The proliferation or migration ability of cells is analyzed by MTS or migration assay. RESULTS: We find that MUC16 and p120ctn are aberrantly overexpressed in 94 clinical OC samples compared with benign ovarian tumors (BOT). MUC16 is a critical inducer of the proliferation and migration of EOC cells and the CTD of MUC16 plays an important role during this process. In addition, we reveal the relationship between MUC16 and p120ctn, which has not previously been studied. We show that MUC16 promotes the translocation of p120ctn to the cytoplasm and consequently activates Rho GTPases to modulate the proliferation and migration abilities of EOC cells. The cell proliferation and migration abilities induced by MUC16 are mediated by p120ctn through RhoA/Cdc42 activation. CONCLUSIONS: The highly expressed MUC16 promotes the translocation of p120ctn to the cytoplasm, where it activates RhoA/Cdc42 to modulate the proliferation and migration abilities of EOC cells. These findings may provide new targets for the treatment of EOC.

Effect of pelvic floor dysfunction on sexual function and quality of life in Chinese women of different ages: An observational study.

AIM: The aim of the present study was to investigate the effect of female pelvic floor dysfunction on sexual function and quality of life among different age groups. METHODS: An observational study was carried out on 648 patients with pelvic organ prolapse (POP) or/and stress urinary incontinence. We assessed female sexual function and quality of life through the Prolapse/Urinary Incontinence Sexual Questionnaire Short Form and Incontinence Quality of Life Scale before surgeries, respectively. Patients were assigned into different age groups. RESULTS: The mean age of all patients was 62.04 ± 9.39 years. A total of 436 patients had POP, 120 patients had stress urinary incontinence and the remaining patients had both. Although sexual frequency decreased with increasing age, 517 patients still remained sexually active. A total of 10.83% of the patients aged ≥70 years were still sexually active, and 8.51% of them had sex less than one time per month. The Prolapse/Urinary Incontinence Sexual Questionnaire Short Form score and sexual frequency decreased with increasing age. However, the Incontinence Quality of Life Scale score increased with increasing age. Patients with only POP and aged ≥70 years showed a higher Incontinence Quality of Life Scale score (P < 0.001). CONCLUSIONS: The present study showed that POP and stress urinary incontinence are common among older Chinese women, and re associated with decreased sexual satisfaction and quality of life in China. It suggests that the existing sexual requirement of these patients should also be reconsidered before surgical assessment. Doctors should take sexual satisfaction into consideration when choosing a personalized type of surgery to improve the patients' quality of life physiologically and psychologically. Geriatr Gerontol Int 2019; 19: 299-304.

Exosomes Released from Tumor-Associated Macrophages Transfer miRNAs That Induce a Treg/Th17 Cell Imbalance in Epithelial Ovarian Cancer.

The immune microenvironment is crucial for epithelial ovarian cancer (EOC) progression and consists of tumor-associated macrophages (TAM) and T lymphocytes, such as regulatory T cells (Treg) and T helper 17 (Th17) cells. In this study, the Treg/Th17 ratio was significantly higher in EOC in situ and in metastatic peritoneal tissues than in benign ovarian tumors and benign peritoneum. The Treg/Th17 ratio was associated with histologic grade and was an independent prognostic factor for overall survival of EOC patients. On the basis of microarray analysis of exosomes derived from TAMs, we identified miRNAs enriched in the exosomes, including miR-29a-3p and miR-21-5p. When the two miRNA mimics were transfected into CD4+ T cells, they directly suppressed STAT3 and regulated Treg/Th17 cells, inducing an imbalance, and they had a synergistic effect on STAT3 inhibition. Taken together, these results indicate that exosomes mediate the interaction between TAMs and T cells, generating an immune-suppressive microenvironment that facilitates EOC progression and metastasis. These findings suggest that targeting these exosomes or their associated miRNAs might pave the way for the development of novel treatments for EOC.

Daily communication, conflict resolution, and marital quality in Chinese marriage: A three-wave, cross-lagged analysis.

Based on three annual waves of data obtained from 268 Chinese couples in the early years of marriage and using a three-wave, cross-lagged approach, the present study examined the associations among daily marital communication, marital conflict resolution, and marital quality. Results indicated unidirectional associations linking daily marital communication or marital conflict resolution to marital quality (instead of reciprocal associations); and when considered simultaneously in a single model, daily marital communication and marital conflict resolution explained variance in marital quality above and beyond each other. Furthermore, the authors also found a significant longitudinal, indirect association linking husbands' daily marital communication at Wave 1 to husbands' marital quality at Wave 3 via husbands' marital conflict resolution at Wave 2. Taken altogether, the current study adds to an emerging body of research aimed at clarifying: (a) the directionality of the associations between couple interactive processes and marital well-being; (b) the unique roles of daily marital communication and marital conflict resolution in predicting marital outcomes; and (c) how daily marital communication and marital conflict resolution may operate in conjunction with each other to shape the development of couple relationship well-being. (PsycINFO Database Record

Impact of transvaginal modified sacrospinous ligament fixation with mesh for the treatment of pelvic organ prolapse-before and after studies.

BACKGROUND: Pelvic organ prolapse (POP) is a common disease in women. The aim of this research was to evaluate the safety, efficacy and complication of transvaginal modified sacrospinous ligament fixation with mesh using for the treatment of vaginal vault prolapse. MATERIALS AND METHODS: This was a prospective study including information from 60 symptomatic women with anterior-apical pelvic floor prolapse. The patients underwent transvaginal modified sacrospinous ligament fixation combined with anterior vaginal wall mesh between May 2014 and Sep 2015. The perioperative data including clinical characteristic, operation time, blood loss, and surgical complications were collected at 1 year and 2 years. During a 2-year follow-up, the primary outcome evaluation included Pelvic organ prolapse Quantification system (POP-Q), Incontinence Quality of Life scale (I-QoL), the Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12) and the Pelvic Floor Distress Inventory, short form 20 (PFDI-20). RESULTS: The mean follow-up time was 2 years (range 24-37 months). The patients' mean age was 66.75 ±â€¯6.44. Of 60 patients who were enrolled in this research, 26 patients had severe stress urinary incontinence (SUI). The mean operation time was 99.14 ±â€¯19.60 min and the mean estimated blood loss was 73.83 ±â€¯41.05 ml. The rate of anatomical success was 98.3% and one patient had a recurrence. The POP-Q point measurements were evaluated preoperatively and postoperatively (P < 0.001). Moreover, the quality of life and sexual activity were all improved postoperatively via I-QoL, PISQ-12 and PFDI-20 scores (P < 0.001). There was no injury to the rectum, bladder, major pelvic vessels and pudendal nerves. However, 18 patients had postoperative complications. CONCLUSIONS: This study showed that transvaginal modified sacrospinous ligament fixation with mesh might be performed easily and might be a safe surgery for elderly patients whose requirements for sexual life were relatively low. Further researches were required to investigate its long-term efficacy.

Crosstalk between TEMs and endothelial cells modulates angiogenesis and metastasis via IGF1-IGF1R signalling in epithelial ovarian cancer.

BACKGROUND: Epithelial ovarian cancer (EOC) is the leading cause of death from gynaecologic malignancies and has a poor prognosis due to metastasis. Drugs targeting the angiogenesis pathway significantly improve patient outcome. However, the key factors linking angiogenesis and metastasis have not been elucidated. In this study, we found Tie2 expressing monocytes (CD14+Tie2+, TEMs) as key contributors to angiogenesis and metastasis of EOC. METHODS: Tissue slides were evaluated by immunofluorescence for the presence of total tissue macrophages and TEMs. The correlation between microvascular density (MVD) values and the TEMs number or ratio was calculated in both ovarian cancer tissues and peritoneum. The rate of TEMs in monocytes was evaluated in the peripheral blood of female healthy donors, benign cysts patients, and EOC patients using flow cytometry. The TEMs rate in ascites from EOC patients was also evaluated by flow cytometry. The concentration of Ang2, as the ligand of Tie2, was examined by ELISA in serum samples of EOC patients, benign cysts patients, and ascites samples of EOC patients. The effects of Ang2 on the migration and the cytokine expression of TEMs were further examined. The pro- angiogenesis activity of TEMs via IGF1 was performed in both in vivo and in vitro. And the IGF1 blocking test was performed using neutralising antibody. RESULTS: TEMs were significantly higher in tumour foci, peripheral blood and ascites in EOC patients. The proportion of TEMs among total tissue macrophages was positively correlated with tumour MVD. In vivo animal results showed that TEMs promoted EOC angiogenesis and metastasis. Further functional and mechanisms studies revealed that concentration of angiopoietin 2 (Ang2), a ligand of Tie2, was elevated in EOC ascites which further recruit TEMs in a dose-dependent manner as a powerful chemokine to TEMs. Recruited TEMs promoted endothelial cell function through IGF1-activated downstream signalling. Blocking secreted IGF1 using inhibiting antibody reduced TEMs mediated angiogenesis and metastasis. CONCLUSIONS: TEMs significantly increased in EOC patients and were recruited to tumour loci by the increased Ang2. The increased TEMs have diagnostic value in ovarian cancer and were positively correlated with the MVD in ovarian cancer tissue. Furthermore, TEMs promote angiogenesis via IGF1 in both in vivo and in vitro experimental systems after stimulation by Ang2. Altogether, this study paves the way to develop novel therapy targets as the axis of Ang2-TEMs-IGF1 in EOC.

Exosomes derived from hypoxic epithelial ovarian cancer deliver microRNA-940 to induce macrophage M2 polarization.

Hypoxia is a common feature of solid tumors. It is closely related to tumor progression. Exosomal microRNAs derived from cancers are considered to be mediators between cancer cells and the tumor microenvironment. In addition, the number of tumor-associated macrophages (TAMs) in the tumor microenvironment has also been demonstrated to correlate with tumor development. However, the relationship between tumor-secreted exosomes and TAM polarization under hypoxic conditions during tumor progression is not clear. Herein, we demonstrated that hypoxia induces the high expression of microRNA-940 (miR­940) in exosomes derived from epithelial ovarian cancer (EOC). We also found that miR­940 is highly expressed in exosomes isolated from ascites of EOC patients. Moreover, the overexpression of miR­940 in macrophages delivered by exosomes stimulated M2 phenotype polarization, while the M2 subtype macrophages promoted EOC proliferation and migration. These results highlight the function of hypoxia in enhancing the high level of expression of miR­940 in tumor exosomes taken up by macrophages. We also showed that the tumor-promoting function of miR­940 is mediated by TAM polarization in EOC. These findings show that tumor-derived exosomal miR­940 induced by hypoxia plays an important role in stimulating TAM polarization in the progression of EOC.

The expression of Duffy antigen receptor for chemokines by epithelial ovarian cancer decreases growth potential.

Epithelial ovarian cancer (EOC) is one of the main causes of cancer-associated mortality in females with gynecological malignancies. Duffy antigen receptor for chemokines (DARC) has previously been reported to be involved in tumor growth and the inhibition of tumor metastasis. However, the association between DARC and EOC remains unknown. The aim of the present study was to investigate the expression of DARC in the SKOV3 human epithelial ovarian cancer cell line with the establishment of a subcutaneous model in nude mice. To investigate the effects of DARC on the tumorigenesis of human epithelial ovarian cancer cells, GV287-DARC-L.V lentiviral vectors containing a DARC overexpression construct were transfected into SKOV3 cells. The present study revealed that transfection with DARC reduced the viability of SKOV3 cells in vitro by performing an MTT assay. SKOV3-DARC and SKOV3-negative control (NC) cells cultured in vitro were injected into nude mice to establish a subcutaneous model. The ovarian tumor volumes and the tumor weights were observed. Immunohistochemistry to detect CD31 expression was used to determine the microvessel density (MVD) in SKOV3-DARC and SKOV3-NC tumors. The results of the present study revealed that DARC-induced inhibition of tumor growth was associated with MVD in xenograft tumors. This suggested that DARC was a negative regulator of tumor growth in EOC, primarily via the inhibition of tumor angiogenesis.

Suppression of endothelial cell migration by tumor associated macrophage-derived exosomes is reversed by epithelial ovarian cancer exosomal lncRNA.

OBJECTIVE: To study the mechanism by which epithelial ovarian cancer (EOC)-derived exosomes restore the migration of endothelial cells that is suppressed by TAM-derived exosomes. METHODS: Exosomes were isolated from TAMs in the ascites of patients with EOC. The effect of exosomes on the expression of endothelial cell miRNA was monitored by PCR. The miRNA mimics were transfected to explore their effects. Microarray data and literature searches were used to predict target genes and the impact of target gene pathways, and small interfering RNA was used to target these genes. We used migration assays to determine whether ovarian cancer cell-derived exosomes participate in the regulation of TAMs and endothelial cells. We used microarray data to identify the target lncRNA, and we constructed target lncRNA expression plasmids to validate targets by Western blotting. RESULTS: We separated TAMs from the ascites of patients with EOC and isolated exosomes from TAM supernatants. After co-culture with HUVECs, these exosomes were efficiently incorporated into HUVECs. The migration of HUVECs was suppressed significantly in the exosome group compared with blank controls (P < 0.05).The miRNA mimic transfection and target gene prediction found that TAM-derived exosomes targeted the miR-146b-5p/TRAF6/NF-κB/MMP2 pathway to suppress endothelial cell migration; this result was supported by PCR and Western blotting analyses. The expression of exosomal miR-146b-5p isolated from serum in the EOC group was significantly increased compared to healthy individuals. Finally, TAM-derived exosomes and EOC SKOV3-derived exosomes in combination stimulated HUVEC cells and overcame the inhibition of endothelial cell migration caused by TAM-derived exosomes. Two lncRNAs that were carried by SKOV3-derived exosomes were identified as NF-κB pathway-associated genes by Western blotting. CONCLUSION: TAM-derived exosomes can inhibit the migration of endothelial cells by targeting the miR-146b-5p/TRAF6/NF-kB/MMP2 pathway. However, EOC-derived exosomes can transfer lncRNAs to remotely reverse this effect of TAMs on endothelial cells.

The expression of asparaginyl endopeptidase promotes growth potential in epithelial ovarian cancer.

Epithelial ovarian cancer (EOC) is the most common and lethal cancer-related death among females in the world. Asparaginyl endopeptidase (AEP) is a member of C13 family peptidases and expressed in the extracellular matrix and tumor cells. The aim of this article is to explore the function of asparaginyl endopeptidase in epithelial ovarian cancer. The expression of AEP was examined in 20 EOC samples, 3 EOC metastasis samples, 6 fallopian tube metastasis samples, 4 peritoneum metastasis samples and 20 benign ovarian tumor samples by immunohistochemistry. The expression of AEP was also evaluated in serum and ascites of EOC patients by elisa. And we used a lentiviral vector to overexpress AEP in human epithelial ovarian cancer cell lines SKOV3ip and detected the function of AEP-SKOV3ip cells both in vitro and in vivo. The growth of AEP-SKOV3ip cells was observed by MTT, migration and tube formation assays in vitro. Additionally, the subcutaneous mice model was used to identify the tumor growth and metastasis in vivo. Mice tumors were stained for CD31 to determine the microvessel density (MVD). We demonstrated that AEP was highly expressed in the EOC patient tissues and ascites. The AEP transfected SKOV3ip cells could both promote tumor growth in vitro and in vivo. The MVD in AEP-SKOV3ip group was higher than that in NC-SKOV3ip group. Therefore, our results demonstrated that AEP could induce EOC growth and progressionboth in vitro and in vivo.

Differential distribution of tumor-associated macrophages and Treg/Th17 cells in the progression of malignant and benign epithelial ovarian tumors.

Epithelial ovarian cancer (EOC) is one of the predominant causes of cancer-associated mortality in women with gynecological oncology. Tumor-associated macrophages (TAMs), regulatory T cells (Treg cells) and T helper cell 17 (Th17) cells have been hypothesized to be involved in the progression of EOC. However, the association between TAMs and T cells remains to be elucidated. The aim of the present study was to investigate the differential distribution of TAMs, Treg cells and Th17 cells in benign ovarian tumor tissues and in tissues from patients with EOC, and to examine their association with the clinical pathology of EOC. A total of 126 tissue samples from patients with EOC and 26 tissue samples from patients with benign ovarian tumors were analyzed, and it was identified that the distribution of TAMs, Treg cells, Th17 cells and the ratio of Treg/Th17 cells were higher in the patients with EOC using triple color immunofluorescence confocal microscopy. The high frequency of TAMs and ratio of Treg/Th17 cells in late tumor grades suggested that they may be significant in tumor progression. The frequency of TAMs was different between the histological types of EOC. Immunohistochemistry was used to investigate the microvessel density (MVD) in the EOC and benign ovarian tumor tissues. A higher MVD was observed in the EOC patient tissues, particularly, in the late tumor grade tissues. The present study provided clinical data demonstrating the high distribution of TAMs and T-cells in EOC, which may contribute to tumor progression through angiogenesis. The mechanisms by which TAMs are associated with Treg cells and Th17 cells requires further investigation as prognostic factors and therapeutic targets for EOC.

Interaction between Treg cells and tumor-associated macrophages in the tumor microenvironment of epithelial ovarian cancer.

Epithelial ovarian cancer (EOC) is the most lethal gynecological malignancy. Inflammatory cells in the EOC microenvironment play a key role in tumor progression. In the present study, we investigated the mechanism of the accumulation of regulatory T cells (Tregs) induced by interleukin-10 (IL-10) derived from tumor-associated macrophages (TAMs) in the EOC microenvironment. The frequency of Tregs and TAMs was detected by immunofluorescence in 40 EOC tissues and 20 benign ovarian tumors, as well as the expression of IL-10 which was assessed by immunohistochemistry. It was found that the frequency of Treg cells and TAMs was significantly higher in the EOC than those in the benign ovarian tumors. The expression of IL-10 was also found to be higher in the EOC than that in the benign tumors. EOC patients with a high frequency of Tregs exhibited a significantly shorter overall survival time compared to those with a low frequency of Tregs. In addition, the expression of IL-10 in ascites and blood serum and the IL-10 released in the co-cultured system supernatants were detected by ELISA. Following CD4+ T-cell co-culturing with macrophages and IL-10, it was observed by flow cytometric analysis that the frequency of Treg cells was increased in the presence of IL-10. It was also established that IL-10 released in the co-cultured supernatants was increased. We also detected the mechanism of Treg cells induced by IL-10 in vivo. The SKOV3 cell tumor volume and weight were much higher in the presence of IL-10 in a mouse subcutaneous model. These data suggest that IL-10 secreted by TAMs increase the frequency of Treg cells through the activation of Foxp3 during T-cell differentiation and promotes tumor progression.

Epithelial ovarian cancer-secreted exosomal miR-222-3p induces polarization of tumor-associated macrophages.

Cancer secreted exosomal miRNAs are emerging as mediators between tumor-stoma crosstalk. Here, we show epithelial ovarian cancer (EOC)-derived exosomes activated macrophages to a tumor-associated macrophage (TAM)-like phenotype with SOCS3/STAT3 pathway involvement, which could facilitate the progression of cancer. MiR-222-3p was enrichment in exosomes released from EOC cells and it could be transferred to macrophages. Overexpression of miR-222-3p in macrophages induced polarization of the M2 phenotype. Luciferase assay verified miR-222-3p targeted SOCS3 genes and expression of SOCS3 was decreased after transfection with a miR-222-3p mimic. Down-regulation of SOCS3 correlated with an increased expression of STAT3 activation. MiR-222-3p could be detected in the exosomes from serum and its levels were related to EOC. These observations propose tumor-derived exosomal miR-222-3p is an effective regulator in the polarization of tumor-promoting M2 macrophages and may be a biomarker of EOC.

Efficacy of Postoperative Hormone Replacement Therapy on Prognosis of Patients with Serous Ovarian Carcinoma.

BACKGROUND: Ovarian cancer is the most common cause of gynecological cancer-associated death. Iatrogenic menopause might adversely affect the quality of life and health outcomes in young female cancer survivors. We evaluated whether postoperative hormone replacement therapy (HRT) had a negative influence on the progression-free survival (PFS) of patients with papillary serous ovarian cancer (SOC). METHODS: We retrospectively reviewed the medical records of patients with papillary SOC, treated from January 1980 to December 2009, who suffered from menopause with or without HRT. Clinical characteristics of patients were compared between the two groups (HRT and non-HRT). Blood samples were collected from all the participants to detect serum cancer antigen (CA) 125. Hazard ratios with 95% confidential intervals for each variable were calculated by univariable and multivariable conditional Logistic regression analyses. RESULTS: Among 112 identified patients, 31 were HRT users and 81 were not. The two groups did not significantly differ in median age at diagnosis (t = 0.652, P = 0.513), International Federation of Gynecology and Obstetrics (FIGO) stage (χ2 = 0.565, P = 0.754), differentiation (χ2 = 1.728, P = 0.422), resection status (χ2 = 0.070, P = 0.791), relapse (χ2 = 0.109, P = 0.741), chemotherapy course (t = -1.079, P = 0.282), follow-up interval (t = 0.878, P = 0.382), or PFS (t = 0.580, P = 0.562). Median Kupperman score at the onset of HRT was 30.81 and 12.19 after the therapy (t = 3.302, P = 0.001). According to the analysis, the strongest independent variables in predicting PFS were FIGO stage and disease that was not optimally debulked. CONCLUSIONS: Postoperative HRT is not a prognostic factor for PFS of patients with papillary SOC. However, multicenter studies are needed to verify and extend our findings.