TGFBI promoter hypermethylation correlating with paclitaxel chemoresistance in ovarian cancer

The purpose of this study is to determine the methylation status of Transforming growth factor-beta-inducible gene-h3 (TGFBI) and its correlation with paclitaxel chemoresistance in ovarian cancer. The methylation status of TGFBI was examined in ovarian cancer and control groups by methylation-specific PCR (MSP) and bisulfite sequencing PCR (BSP). The TGFBI expression and cell viability were compared by Quantitative Real-Time PCR, Western Blotting and MTT assay before and after demethylating agent 5-aza-2'-deoxycytidine (5-aza-dc) treatment in 6 cell lines (SKOV3, SKOV3/TR, SKOV3/DDP, A2780, 2780/TR, OVCAR8). In our results, TGFBI methylation was detected in 29/40 (72.5%) of ovarian cancer and 1/10 (10%) of benign ovarian tumors. No methylation was detected in normal ovarian tissues (P < 0.001). No statistical correlation between RUNX3 methylation and clinicopathological characteristics was observed. A significant correlation between TGFBI methylation and loss of TGFBI mRNA expression was found (P < 0.001). The methylation level of TGFBI was significantly higher in paclitaxel resistant cell lines (SKOV3/TR and 2780/TR) than that in the sensitive pairs (P < 0.001). After 5-aza-dc treatment, the relative expression of TGFBI mRNA and protein increased significantly in SKOV3/TR and A2780/TR cells. However, no statistical differences of relative TGFBI mRNA expression and protein were found in other cells (all P > 0.05), which showed that re-expression of TGFBI could reverse paclitaxel chemoresistance. Our results show that TGFBI is frequently methylated and associated with paclitaxel-resistance in ovarian cancer. TGFBI might be a potential therapeutic target for the enhancement of responses to chemotherapy in ovarian cancer patients

Phase separation and enhanced wear resistance of Cu88Fe12 immiscible coating prepared by laser cladding

In order to eliminate the microstructure segregation of Cu–Fe immiscible alloys which characterized with a liquid miscible gap, the Cu88Fe12 (wt.%) immiscible coating was prepared by laser cladding. The phase separation characteristic and wear resistance of the Cu88Fe12 (wt.%) immiscible coating were also investigated. The results show that the size of the milled Cu88Fe12 composite powder is reduced comparing to that of the un-milled powder due to fracturing during mechanical milling. Moreover, the demixing or delamination disappears in the Cu88Fe12 immiscible coating and a large amount of face-centered-cubic (fcc) γ-Fe and body-centered-cubic (bcc) α-Fe particles are dispersed in the face-centered-cubic (fcc) ɛ-Cu matrix as a result of liquid phase separation. The size of Fe-rich particles presents an increasing tendency from the bottom to the top of the immiscible coating. As a result, the microhardness of the immiscible coating is improved compared with brass (∼138 HV0.2) due to the presence of high-hardness Fe-rich particles (∼191 HV0.2) and the solid solution strengthening effect of Fe in Cu-rich matrix. Furthermore, the width of ploughing, the width and height of wear scar on the surface of the immiscible coating are much less than those on the surface of brass. Therefore, the wear resistance of the immiscible coating is remarkably enhanced compared with brass

Blockage of transdifferentiation from fibroblast to myofibroblast in experimental ovarian cancer models

<p>Abstract</p> <p>Background</p> <p>Tumour stromal myofibroblasts can promote tumour invasion. As these cells are genetically more stable than cancer cells, there has been enormous interest in developing targeted molecular therapies against them. Chloride intracellular channel 4 (CLIC4) and reactive oxygen species (ROS) have been linked with promoting stromal cell transdifferentiation in various cancers, but little is known of their roles in ovarian cancer. In this study, we examined the functional roles that both CLIC4 and ROS play in the process of ovarian cancer cell-stimulated or TGF-β1 induced fibroblast-to-myofibroblast transdifferentiation. We also examine whether it is possible to reverse such a process, with the aim of developing novel therapies against ovarian cancer by targeting activated transdifferentiated myofibroblasts.</p> <p>Results</p> <p>We demonstrate that TGF-β1 induced or CM^SKOV3activate transdifferentiated myofibroblasts (fibroblasts). These fibroblasts mimic "reactive" stromal myofibroblasts and demonstrate significant up-regulation of CLIC4 expression and increased level of ROS production. Blocking the production of ROS with an antioxidant consequently reduces the expression of CLIC4, and is accompanied by disappearance of <it>α</it>-smooth-muscle actin (α-SMA), a myofibroblast marker, suggesting ROS acts as a signalling molecule that promotes and enhances CLIC4 activities in the myofibroblast transdifferentiaton process. Down-regulation of CLIC4 with a generic agent or specific siRNA both significantly reduces the expression of factors related to the phenotypes and functions of myofibroblasts, such as α-SMA, hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF), thus reversing the myofibroblast phenotype back to fibroblasts. These results convincingly show that ROS and CLIC4 are responsible for TGF-β1 induced fibroblast-to-myofibroblast transdifferentiaton and down-regulation of both is sufficient to block transdifferentiated myofibroblasts.</p> <p>Conclusion</p> <p>Molecular targeting of ROS and CLIC4 has the potential to develop novel therapies for ovarian cancer.</p

Starch-Based Flexible Coating for Food Packaging Paper with Exceptional Hydrophobicity and Antimicrobial Activity

Herein, we fabricated a starch-based flexible coating for food packaging papers with excellent hydrophobicity and antimicrobial properties. FTIR (Fourier transform infrared) and XRD (X-ray diffraction) spectra revealed the homogeneous dispersion of the ZnO nanoparticles (NPs) in the composite film within 5% ZnO NP dosage. SEM (scanning electron microscope) and AFM (atomic force microscope) micrographs confirmed the increased roughness on the composite film with the increased dosages of ZnO NPs. Hydrophobic characteristics showed that dramatic enhancement was obtained in the values and stabilities of DCAs (dynamic contact angles) in the resultant film and coated paper. TG (thermogravimetry) results demonstrated the increased thermal stabilities of the composite films. Significantly, a decreased water vapor transmission rate was observed in the coated paper. When 20% guanidine-based starch and 2% CMC (carboxy methyl cellulose) was added, a flexible coating with excellent antimicrobial activity towards Escherichia coli can be obtained. Furthermore, the migration of ZnO NPs into the food simulants was well below the overall migration legislative limit. The resultant starch-based flexible composite film and coated paper established an effective approach to develop a green-based material for food packaging applications

A single center survey of anxiety and depression status in Chinese cystic fibrosis patients

Objective To study anxiety and depression in Chinese patients with cystic fibrosis (CF). Methods Clinical data from 28 CF patients in the cystic fibrosis registry study of Peking Union Medical College Hospital were collected and analyzed. Hospital anxiety and depression scale was used to identify anxiety and depression. Results The cohort had 42.9% (12/28) males with a mean age of 23±4.8 years. 14 patients (50%) had anxiety or depression. Among them, 8 subjects (28.6%) had moderate and severe anxiety or depression, and 5 patients (17.9%) were with moderate and severe anxiety, and 7 patients(25%) with moderate and severe depression. One patient(3.6%) had severe anxiety, no patients with severe depression, and four patients (14.3%) had both moderate and severe anxiety and depression. Compared to patients without anxiety or depression, patients with anxiety had significantly lower FVC% pred (54.5%±18.8% with anxiety, 80.0%±14.6% without anxiety or depression, P＜0.05), and patients with depression had significantly lower FEV1% pred (41.6%±21.6% with depression, 64.2%±20.9% without anxiety or depression, P＜0.05) and FVC% pred (53.9%±21.0% with depression, 80.0%±14.6% without anxiety or depression, P＜0.01). Conclusions Half of CF patients have anxiety or depression, with a high comorbidity rate of anxiety and depression and a low prevalence of severe anxiety or depression in our cohort. CF patients with anxiety or depression have worse pulmonary ventilationy function compared to CF patients without anxiety or depression

Seed soaking with melatonin promotes seed germination under chromium stress via enhancing reserve mobilization and antioxidant metabolism in wheat

Chromium (Cr) pollution has serious harm to crop growth, while little is known on the role of melatonin (MT) on seed germination and physiology in Cr-stressed wheat. The effects of seed soaking with MT on growth, reserve mobilization, osmotic regulation and antioxidant capacity of wheat seeds during germination under hexavalent chromium (100 μM) stress were investigated. The results indicated that Cr toxicity decreased the seed germination rate by 16% and suppressed the growth of germinated seeds compared to unstressed seeds. MT in the concentration-dependent manner increased germination rate and promoted subsequent growth when seeds were exposed to Cr stress, but the effect could be counteracted at high concentration. Seed soaking with MT (100 μM) markedly decreased Cr accumulation in seeds, radicals and coleoptiles by 15%, 6% and 15%, respectively, and enhanced α-amylase activity and soluble sugar and free amino acids content in seeds to improve reserve mobilization under Cr stress, compared with Cr treatment. Furthermore, decreasing the level of osmotic regulators (soluble sugar and soluble protein) in radicles under MT combined with Cr treatment confirmed the reduction of osmotic stress caused by Cr stress. Importantly, MT pretreatment reduced H2O2 content by 19% and O2·− release rate by 45% in radicles under Cr toxicity compared with Cr-stressed wheat, in terms of promoting scavenging ability and decreasing production ability, which was to upregulate the activities and encoding genes expression levels of superoxide dismutase (SOD), catalase (CAT), ascorbic acid peroxidase (APX) and peroxidase (POD) and to downregulate plasma membrane-bound NADPH oxidase (NOX) encoding genes (TaRbohD, TaRbohF) expression, respectively. In all, these results provided evidence that seed soaking with MT could be a potentially method to protect wheat seeds from Cr toxicity, which effectively ameliorated germination under Cr stress by enhancing reserve mobilization and antioxidant metabolism in wheat

Membranous and nuclear staining of CLDN18 in HPV‐independent and HPV‐associated endocervical adenocarcinomas

Abstract Objectives A classification system for endocervical adenocarcinoma (ECA) based on high‐risk human papillomavirus (HPV) status has been established; however, the immunohistochemical markers distinguishing HPV‐independent and HPV‐associated ECAs have not been fully described. Here, we aimed to characterize ECA immunopathological features. Methods We evaluated the immunohistochemical profile of CLDN18, CDX2, PAX8, p16, p53, and CEA in 60 ECAs comprising 10 HPV‐independent ECAs and 50 HPV‐associated ECAs. Both the membranous and nuclear expression levels of CLDN18 were analyzed. Results Membranous CLDN18 (CLDN18 [M]) was found to be expressed in the mucinous epithelium of all HPV‐independent ECAs, including eight gastric‐type ECAs (G‐ECAs), one endometrioid ECA, and one clear cell ECA, but no nuclear CLDN18 (CLDN18 [N]) expression was detected in HPV‐independent ECAs. Among HPV‐associated ECAs, CLDN18 (M) expression levels in intestinal‐type (I‐ECAs) and usual‐type ECAs (U‐ECAs) were significantly different from those in invasive stratified mucin‐producing (iSMILE) carcinomas (p = 0.036). Positive CLDN18 (M) staining was present in 55.6% (5/9) of intestinal‐type and 39.4% (13/33) of usual‐type ECAs and was not present in iSMILE ECAs. Silva pattern C cancers expressed higher levels of CLDN18 (M) than Silva pattern A and B cancers (p = 0.004), whereas the CLDN18 (N) expression levels in cancers showing Silva pattern A were significantly higher than those in cancers exhibiting Silva patterns B and C (p < 0.001). Conclusion Membranous CLDN18 is expressed in ECAs and is particularly frequently expressed in HPV‐independent ECAs, and membranous CLDN18 expression has potential as a therapeutic target. Nuclear staining of CLDN18 is a new immunohistochemical marker for diagnosing Silva pattern A HPV‐associated ECAs and is associated with a good prognosis. Further studies should investigate the therapeutic and prognostic significance of membranous and nuclear CLDN18 expression and develop a related test that can be implemented in the clinical evaluation of ECAs