A highly sensitive and rapid method of in-situ surface-enhanced Raman spectroscopy (SERS) combining with electrochemical preconcentration (EP) in detecting malachite green (MG) in aquaculture water ...was established. Ag nanoparticles (AgNPs) were synthesized and spread onto the surface of gold electrodes after centrifuging to produce SERS-active substrates. After optimizing the pH values, preconcentration potentials and times, in-situ EP-SERS detection was carried out. A sensitive and rapid analysis of the low-concentration MG was accomplished within 200s and the limit of detection was 2.4 × 10−16M.
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•The method of in-situ SERS-EP in detecting MG was established firstly.•The SERS substrate is fast, simple to be prepared as well as at low cost.•The EP-SERS for MG detection has higher detection limit compared to earlier methods.•The whole EP-SERS process was timesaving and efficient.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
Epithelial ovarian cancer (EOC) is the most lethal of the gynecological malignancies. Exploring the molecular mechanisms and major factors of invasion and metastasis could have great significance for ...the treatment and prognosis of EOC. Studies have demonstrated that microRNA 106b (miR-106b) may be a promising therapeutic target for inhibiting breast cancer bone metastasis, but the role of miR-106b in EOC is largely unknown. In this work, miRNA-106b expression was quantified in various ovarian tissues and tumors. Ovarian carcinoma cell lines were transfected with miR-106b, after which, cell phenotype and expression of relevant molecules was assayed. Dual-luciferase reporter assays and xenograft mouse models were also used to investigate miR-106b and its target gene. MiR-106b mRNA expression was found to be significantly higher in normal ovarian tissues and benign tumors than in ovarian carcinomas and borderline tumors (p < 0.01), and was negatively associated with differentiation (Well vs. Por & Mod) and the International Federation of Gynecology and Obstetrics (FIGO) staging (stage I/II vs. stage III/IV) in ovarian carcinoma (p < 0.05). MiR-106b transfection reduced cell proliferation; promoted G1 or S arrest and apoptosis (p < 0.05); suppressed cell migration and invasion (p < 0.05); reduced Ras homolog gene family member C (RhoC), P70 ribosomal S6 kinase (P70S6K), Bcl-xL, Matrix metallopeptidase 2 (MMP2), MMP9 mRNA and protein expression; and induced p53 expression (p < 0.05). Dual-luciferase reporter assays indicated that miR-106b directly targets RhoC by binding its 3'UTR. MiR-106b transfection also suppressed tumor development and RhoC expression in vivo in xenograft mouse models. This is the first demonstration that miR-106b may inhibit tumorigenesis and progression of EOC by targeting RhoC. The involvement of miR-106b-mediated RhoC downregulation in EOC aggression may give extended insights into molecular mechanisms underlying cancer aggression. Approaches aimed at overexpressing miR-106b may serve as promising therapeutic strategies for treating EOC patients.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Abstract
The detection of microbial infections by plants induces the rapid formation of immune receptor complexes at the plasma membrane. However, how this process is controlled to ensure proper ...immune signaling remains largely unknown. Here, we found that the Nicotiana benthamiana membrane-localized leucine-rich repeat receptor-like kinase BAK1-INTERACTING RLK 2 (NbBIR2) constitutively associates with BRI1-ASSOCIATED RECEPTOR KINASE 1 (NbBAK1) in vivo and in vitro and promotes complex formation with pattern recognition receptors. In addition, NbBIR2 is targeted by 2 RING-type ubiquitin E3 ligases, SNC1-INFLUENCING PLANT E3 LIGASE REVERSE 2a (NbSNIPER2a) and NbSNIPER2b, for ubiquitination and subsequent degradation in planta. NbSNIPER2a and NbSNIPER2b interact with NbBIR2 in vivo and in vitro and are released from NbBIR2 upon treatment with different microbial patterns. Furthermore, accumulation of NbBIR2 in response to microbial patterns is tightly associated with NbBAK1 abundance in N. benthamiana. NbBAK1 acts as a modular protein that stabilizes NbBIR2 by competing with NbSNIPER2a or NbSNIPER2b for association with NbBIR2. Similar to NbBAK1, NbBIR2 positively regulates pattern-triggered immunity and resistance to bacterial and oomycete pathogens in N. benthamiana, whereas NbSNIPER2a and NbSNIPER2b have the opposite effect. Together, these results reveal a feedback regulatory mechanism employed by plants to tailor pattern-triggered immune signaling.
Nicotiana benthamiana fine-tunes pattern-triggered immunity through the receptor-like kinase NbBIR2, the co-receptor NbBAK1, and the E3 ligases NbSNIPER2a and NbSNIPER2b.
Abstract
Encouraged by the goal of achieving net-zero carbon emission by 2050, the exploration of insulating material becomes one of the core projects to the electrical equipment innovations. The ...rise of composites enables the insulating material to be endowed with more functions. By this concern, adopting the coupling agents is one of the effective methods to improve the compatibility of composites, which highly strengthens the desired properties. The functional groups of coupling agents, nevertheless, could not always show a positive impact to access enhanced properties. In this work, four types of silane coupling agents including amino-propyl, glycidyletheroxy-propyl (GP), vinyl, and methacryloxy-propyl functional groups are employed to modify the aluminum nitride (AlN) fillers, and the modification effects on the insulating and thermal conductivity of the AlN/poly(m-phenylenedicarbonyl-m-phenylenediamine) (PMIA) composite paper are systematically investigated and compared. The results show that a proper coupling agent is beneficial to the uniform dispersion of the inorganic filler in the organic matrix and highly contributes to their interface quality, where the heat transfer path is established that boosts the heat dissipation. By tailoring the trap depth and density toward deeper and higher, the carrier transport is highly confined which enhances the breakdown strength to a large extent. Therefore, high breakdown strength and thermal conductivity of 182.9 kV mm
−1
and 0.302 W (m·K)
−1
, respectively, are achieved in the GP modified AlN/PMIA paper, which are 16.7% and 167.4% higher than that of pure PMIA paper.
To study the earth pressure characteristics of EPS particles light weight soil, the change laws of coefficient of earth pressure at rest are analyzed by
K
0
consolidation tests. The results show that ...the measured coefficient of earth pressure at rest of light weight soil is about 0.14–0.31, which is only 1/3–1/2 of the theoretical calculation values of Jaky formula. The measured coefficient of earth pressure at rest of remolded soil is 3/4 of the theoretical calculation values of Jaky formula.When the EPS particles volume ratio and cement mixed ratio of light weight soil are constant respectively, the coefficient of earth pressure at rest first increases and finally tends to be a stable value with the increasing of vertical stress. In the unloading process, the coefficient of earth pressure at rest first increases slowly and then increases rapidly with the decreasing of vertical stresses. When EPS particles volume is larger, the coefficient of earth pressure at rest is greater than 1 during unloading process. During the loading and unloading process, the loading curve and unloading curve are quite different, and the unloading curve is above the loading curve. When the unloading is completed, the residual lateral stress of light weight soil increases with the increasing of EPS particles volume ratios and the decreasing of cement mixed ratios. When the vertical stress is constant, the lateral stress and the coefficient of earth pressure at rest increase with the increasing of EPS particles volume ratios and the decreasing of cement mixed ratios.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Recent studies have shown that microRNAs may regulate the ABCB1 gene (ATP-binding cassette, sub-family B MDR/TAP, member 1). Computational programs have predicted that the 3'-untranslated region ...(3'-UTR) of ABCB1 contains a potential miRNA-binding site for miR-186. Here, we investigated the role of miR-186 in sensitizing ovarian cancer cells to paclitaxel and cisplatin.
Human ovarian carcinoma cell lines OVCAR3, A2780, A2780/DDP, and A2780/Taxol were exposed to paclitaxel or cisplatin with or without miR-186 transfection, and cell viability was determined by MTT assay. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis were used to assess the MDR1, GST-π, and MRP1 expression levels. Dual-luciferase reporter assay was used to reveal the correlation between miR-186 and ABCB1. Lower miR-186 while higher MDR1 and GST-π mRNA expression levels were found in the A2780/Taxol and A2780/DDP cells than in the A2780 cells. After miR-186 transfection, all the cell lines showed increased sensitivity to paclitaxel and cisplatin. MiR-186 transfection induced apoptosis while anti-miR-186 transfection reduced apoptosis. The dual-luciferase reporter assay verified that that miR-186 combined with the 3'-untranslated region (UTR) of ABCB1. MDR1 and GST-π mRNA and protein expression levels were downregulated after transfection with miR-186 but upregulated following anti-miR-186 transfection compared to the mock and negative control cancer cells; however, the MRP1 expression levels did not significantly differ among the groups.
Our results are the first to demonstrate that miR-186 may sensitize ovarian cancer cell to paclitaxel and cisplatin by targeting ABCB1 and modulating the expression of GST-π.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Metastasis‐associated in colon cancer‐1 (MACC1), has recently been identified as a key regulator in the progression of many cancers. However, its role in endometrial carcinoma (EC) remains unknown. ...MACC1 expression was determined in EC and normal endometrial tissues by immunohistochemistry. EC cell phenotypes and related molecules were examined after MACC1 downregulation by Small interfering RNA (siRNA) or microRNA (miRNA) transfection. We found that MACC1 was highly expressed in EC tissues than normal samples, and was significantly different in FIGO staging (I and II vs. III and IV), the depth of myometrial infiltration (<1/2 vs. ≥1/2), lymph nodes metastasis (negative vs. positive), besides, MACC1 overexpression was correlated with lower cumulative and relapse‐free survival rate. MACC1 downregulation by siRNA transfection significantly induced G1 phrase arrest, suppressed EC cell proliferation, migration, and invasion. In addition, MACC1 downregulation also reduced expression of Cyclin D1 and Cyclin‐dependent Kinase 2 (CDK2), N‐cadherin (N‐Ca), α‐SMA, matrix metalloproteinase 2 (MMP2), and MMP9, but increased expression of E‐cadherin (E‐Ca). Bioinformatic predictions and dual‐luciferase reporter assays indicate that MACC1 is a possible target of miR‐23b. MiR‐23b overexpression reduced MACC1 expression in vitro and induced G1 phrase arrest, suppressed cell proliferation, migration, and invasion. MiR‐23b transfection also reduced Cyclin D1 and CDK2, N‐Ca, α‐SMA, MMP2, MMP9 expression, but increased E‐Ca expression. Furthermore, the nude mouse xenograft assay showed that miR‐23b overexpression suppressed tumour growth through downregulating MACC1 expression. Taken together, our results demonstrate for the first time that MACC1 may be a new and important diagnosis and therapeutic target of endometrial carcinoma.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
The assemblies of plasmonic nanoparticles (NPs) are the universal methods for enhancing their surface-enhanced Raman scattering (SERS) activities. However, the present methods suffer from the ...problems of poor reproducibility, complicated fabrication, or the adsorption of ligands on the surface, which limit their practical applications. In this work, by using a facile freeze-thaw method, we are able to fabricate the assemblies of Ag NPs with highly reproducible SERS activity without the use of ligands. Moreover, the Ag NPs can be well kept in a frozen state for a long time with few influences on the reproducibility (relative standard deviation, RSD ca. 7%), while those kept in colloid (4 °C) suffer from gradual surface oxidation and aggregation. Such a simple freeze-thaw method does not require the introduction of any ligands (or linkers) with long-term stability and reproducibility, implying its wide applications in practical SERS sensing.
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IJS, KILJ, NUK, PNG, UL, UM
As one of the most frequently diagnosed cancers in women, the development and progression of epithelial ovarian carcinoma (EOC) remains an open area of research. The role of long non‐coding RNAs ...(lncRNAs) in EOC is an emerging field of study. We found that LncRNA TDRG1 (human testis development‐related gene 1) was highly expressed in EOC tissues than in normal ovarian tissues, and expression differed significantly with differentiation. LncRNA TDRG1 downregulation suppressed EOC cell proliferation, migration, and invasion, while its overexpression had the opposite effect. Bioinformatic predictions and dual‐luciferase reporter assays showed that LncRNA TDRG1 has possible miRNA‐93 (miR‐93) binding sites. LncRNA TDRG1 downregulation upregulated miR‐93 expression, while its overexpression reduced miR‐93 expression. In addition, TDRG1 downregulation reduced the expression of Ras homolog gene family member C (RhoC), P70 ribosomal S6 kinase (P70S6 K), Bcl‐xL, and matrix metalloproteinase 2 (MMP2) protein, which are regulated by miR‐93, while its upregulation induced RhoC, P70S6 K, Bcl‐xL, and MMP2 protein expression. In vivo, LncRNA TDRG1 overexpression induced tumor development and RhoC expression. Taken together, our results demonstrated for the first time that LncRNA TDRG1 may be a new and important diagnostic and therapeutic target in EOC.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
AIM:To investigate the expression of key biomarkers in hepatoma cell lines,tumor cells from patients’blood samples,and tumor tissues.METHODS:We performed the biomarker tests in two steps.First,cells ...plated on coverslips were used to assess biomarkers,and fluorescence intensities were calculated using the NIH Image J software.The measured values were analyzed using the SPSS19.0 software to make comparisons among eight cell lines.Second,eighty-four individual samples were used to assess the biomarkers’expression.Negative enrichment of the blood samples was performed,and karyocytes were isolated and dropped onto pretreated glass slides for further analysis by immunofluorescence staining.Fluorescence intensities were compared among hepatocellular carcinoma(HCC)patients,chronic HBV-infected patients,and healthy controls following methods similar to those used for cell lines.The relationships between the expression of biomarkers and clinical pathological parameters were analyzed by Spearman rank correlation tests.In addition,we studied the distinct biomarkers’expression with three-dimensional laser confocal microscopy reconstructions,and Kaplan-Meier survival analysis was performed to understand the clinical significance of these biomarkers.RESULTS:Microscopic examination and fluorescence intensity calculations indicated that cytokeratin 8/18/19(CK)expression was significantly higher in six of the seven HCC cell lines examined than in the control cells,and the expression levels of asialoglycoprotein receptor(ASGPR)and glypican-3(GPC3)were higher in all seven HCC cell lines than in the control.Cells obtained from HCC patients’blood samples also displayed significantly higher expression levels of ASGPR,GPC3,and CK than cells from chronic HBV-infected patients or healthy controls;these proteins may be valuable surface biomarkers for identifying HCC circulating tumor cells isolated and enriched from the blood samples.The stem cell-like and epithelial-mesenchymal transition-related biomarkers could be detected on the karyocyte slides.ASGPR and GPC3 were expressed at high levels,and thus three-dimensional reconstructions were used to observe their expression in detail.This analysis indicated that GPC3 was localized in the cytoplasm and membrane,but that ASGPR had a polar localization.Survival analyses showed that expression of GPC3 and ASGPR is associated with a patient’s overall survival(OS).CONCLUSION:ASGPR,GPC3,and CK may be valuable HCC biomarkers for CTC detection;the expression of ASGPR and GPC3 might be helpful for understanding patients’OS.
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