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In mammals, nemo-like kinase 2 (NLK2) is a conservative protein kinase involved in Wnt/β-catenin signaling pathway and immune response. However, the role of NLK2 in immune response in teleost remain unclear. In this study, we identified an ortholog of mammalian NLK from grass carp (Ctenopharyngodon idellus) named CiNLK2. CiNLK2 shares a high level of homology with the counterparts, especially with that of Cyprinus carpio. CiNLK2 was ubiquitously expressed in all tested tissues (liver, brain, spleen, gill, kidney and eye) and its expression was up-regulated under the treatment with poly I:C or GCRV. Overexpression of CiNLK2 suppressed the production of IFN I in CIK cells whether or not treated with poly I:C. However, knockdown of CiNLK2 increased the expression level of IFN I. The analysis of subcellular localization showed that CiNLK2 protein was scattered throughout the cytoplasm and nucleus. In terms of mechanism, CiNLK2 can directly interact with MAVS and inhibit MAVS-induced IFN I response. Moreover, CiNLK2 increased the phosphorylation level of MAVS, which led to the degradation of MAVS protein. On the other hand, CiNLK2 suppressed the phosphorylation and nuclear translocation of IRF3. In general, CiNLK2 served as an inhibitor for IFN I response by targeting MAVS-IRF3 signal axis.

In mammals, DYRK2 increases p53 phosphorylation level by interacting with it and then promotes cell apoptosis. However, the function of fish DYRK2 has not yet been elucidated. In this paper, we cloned and identified the coding sequence (CDS) of a grass carp DYRK2 (CiDYRK2) which is 1773 bp in length and encodes 590 amino acids. SMART predictive analysis showed that CiDYRK2 possesses a serine/threonine kinase domain. Subsequently, we used the dsRNA analog polyinosinic-polycytidylic acid (poly (I:C) and Grass carp reovirus (GCRV) to stimulate grass carp and CIK cells for different times and found that CiDYRK2 mRNA was significantly up-regulated both in fish tissues and cells. To explore the function of CiDYRK2, we carried out overexpression and knockdown experiments of CiDYRK2 in CIK cells. Real-time quantitative PCR (Q-PCR), TdT-mediated dUTP nick end labeling (TUNEL) assay and flow cytometry were used to detect the ratio of BAX/BCL-2 mRNA, the number of TUNEL positive cells, the proportion of Annexin V-positive cells respectively. The results showed that CiDYRK2 significantly up-regulated BAX/Bcl-2 mRNA ratio and increased the number of TUNEL-positive cells, as well as the proportion of Annexin V-positive cells. On the contrary, knock-down of CiDYRK2 significantly down-regulated BAX/Bcl-2 mRNA ratio in the cells. Therefore, CiDYRK2 promoted cell apoptosis. To study the molecular mechanism by which CiDYRK2 promoting cell apoptosis, subcellular localization and immunoprecipitation experiments were used to study the relationship between grass carp DYRK2 and the pro-apoptotic protein p53. The results showed that CiDYRK2 and Cip53 were located and co-localized in the nucleus. Co-immunoprecipitation experiment also showed that CiDYRK2 and Cip53 can bind with each other. We further found that DYRK2 can increase the phosphorylation level of p53. In a word, our results showed that grass carp DYRK2 induces cell apoptosis by increasing the phosphorylation level of p53.

As a NAD+-dependent deacetylase, SIRT1 is widely involved in apoptosis and cellular inflammation via multiple pathways such as p53, NF-кB and STAT. More and more studies have shown that p53 is the first non-histone deacetylation target of SIRT1. SIRT1-p53 axis thus plays an important role in mammalian cells. IRF9 is an important member of interferon regulator factor family and performs an important role in innate immunity against foreign virus invasion. More importantly, human IRF9 can suppress the SIRT1-p53 axis. However, the functions and relationship between IRF9 and SIRT1-p53 axis are rarely studied in fish. To this end, we made a preliminary research on the functions of grass carp (Ctenopharyngodon idella) IRF9, SIRT1 and p53 in apoptosis and innate immunity. Firstly, we cloned and identified the ORF of SIRT1 (named CiSIRT1, MN125614) from C. idella and demonstrated that CiIRF9 promoted apoptosis, while CiSIRT1 inhibited apoptosis by flow cytometry and TUNEL experiments. Next, we found the interaction between CiSIRT1 and Cip53 in vivo by co-immunoprecipitation experiments. Moreover, the colocalization analysis also showed CiSIRT1 and Cip53 were mainly distributed in nucleus. Thirdly, we got a conclusion that CiIRF9 can repress the expression of CiSIRT1, implying that CiIRF9 regulates CiSIRT1-p53 axis. Finally, CiSIRT1 mRNA level was significantly up-regulated and the expression reached the highest level at 24 h post poly (I:C) stimulation in CIK cells. So, CiSIRT1 may exert an important function in innate immunity. Furthermore, we found CiSIRT1 down-regulated the expression of CiIFN1. In summary, CiIRF9 promotes apoptosis and innate immunity by inhibiting SIRT1-p53 axis. These findings will provide a new theoretical basis for the research on teleost innate immunity.

Accurate diagnosis of Mild Cognitive Impairment (MCI) has gained much more attention in the past few years. It should be noted that, in many recent works, the brain connection estimated by fMRI data have been provided the effective and robust biomarker for several neurological-disorders diagnosis. However, the existing works only focus on the single connection pattern (e.g., Pearson’s Correlation) for diagnosis, which often ignores the information from other patterns for diagnosis. Consequently, such approaches may not be sufficient to reveal the underlying connection differences between the groups of disease-affected patients and normal controls (NC), which limited its performance. As a result, Multiple Connection Pattern Combination (MCPC) method from Single Modal Data is come into beings. Specifically, we propose to combine three patterns of connections including Pearson’s Correlation, Partial Correlation and Dynamic Casual Modelling to identify MCI from Normal Controls (NC), using a kernel combination trick. 68 MCI and 69 NC from ADNI dataset are used for development and validation of our proposed MCPC method. As a result, the proposed methods achieved 87.40 % classification accuracy, significantly outperformed the case of methods using the single connection pattern.

It is critical for human health to develop sensitive and rapid analytical methods for detecting doxycycline (DOX) residues in food. This paper presents a novel metal-organic framework nanomaterial (Zn-MOF) based on dithiodiglycolic acid and its application in DOX detection by fluorescent probe method. Zn-MOF itself does not fluoresce. When DOX is added, the system exhibits strong fluorescence (100-fold) at 530 nm. The fluorescence intensity displayed an excellent linear relationship with DOX concentration with a detection limit of 2.7 nM. The reaction solution's fluorescence displayed a visible color shift from colorless to yellow that was concentration-dependent. A smartphone was used to detect DOX by recognizing the red, green, and blue values of the reaction solution and the corresponding test paper. The use of smartphones can speed up the detection process and streamline operations, offering a sensitive and visible method for the quantitative detection of DOX residues in actual samples. Interestingly, Zn-MOF can discriminate DOX from other tetracyclines with high selectivity. This material has been used successfully as a fluorescent probe to determine DOX in fish samples with an average spiked recovery of 94.6 % ∼ 95.1 %. The DOX levels in the measured perch samples were 1.25 ∼ 157 μg/kg. There are 2 batches of DOX exceeding the standard in 14 batches.

The paper investigates a non-intrusive parallel time integration with multigrid for space-fractional diffusion equations in two spatial dimensions, which is discretized by the space–time finite element method to propagate solutions. We develop a multigrid-reduction-in-time (MGRIT) algorithm with time-dependent time-grid propagators and provide its two-level convergence theory under the assumptions of the stability and simultaneous diagonalizability on time-grid propagators. Numerical results show that the proposed method possesses the saturation error order, theoretical results of the two-level variant deliver good predictions for our model problems, and significant speedups of the MGRIT can be achieved when compared to the two-level variant with F-relaxation (an equivalent version of the parareal algorithm) and the sequential time-stepping approach.

Hypothesis: Polycarboxylate ether (PCE) comb-copolymers are widely used as water reducing agents in the concrete industry while maintaining a high fluidity via the polymer adsorption to the cement particles. PCE copolymers with a broad range of structures are well established by Free radical polymerization, however, understanding the structure-property relationship is still complex due to the high polydispersity of PCE copolymers prepared by conventional polymerization. The influence of different structural parameters using well-defined polymeric structures is yet to be explored. Experiments: In this study, two different types of comb-like random copolymers, namely polycarboxylate ether (PCE; poly(oligo(ethylene glycol) methyl ether methacrylate/methacrylic acid)) and polysulfonate ether (PSE; poly(oligo(ethylene glycol) methyl ether acrylate/sodium 4-styrenesulfonate)), were synthesized by RAFT polymerization to enable the synthesis of polymers with controlled features. The effect of charge types and side chain lengths on the adsorption, rheology, and dispersing ability of cement pastes have been studied. Findings: RAFT polymerization could be used to prepare PCE random copolymers with good control over the polymer molecular weight and narrow polydispersity (D < 1.3). Results revealed that the zeta-potential values depend on both the charge type and side chain lengths. Copolymers containing SO3- exhibited higher absolute negative zeta-potential values than COO- while PCE copolymers with shorter side chains developed higher absolute negative zeta-potential values. On the other hand, the adsorption study demonstrated that decreasing the side chain lengths lead to higher adsorption of PCE copolymers while Copolymers with COO- groups were found to be adsorbed more than SO3- counterparts. These results are further confirmed with the rheological studies and it is found that the shorter the side chain, the lower the yield stress and the higher the dispersion of cement pastes but to a limited effect. Additionally, the charge types have a major influence on the performance of superplasticizers. This study could make further progress in establishing superplasticizers with controlled architectures for better performance.

In mammals, toll-like receptor 3 (TLR3) is capable of recognizing double-stranded RNA and then initiates transcription of IFN-β. TLR3 can activate the innate immune system by phosphorylating extracellular signal-regulated kinase 1 (ERK1) in the mitogen-activated protein kinase (MAPK) pathway. As a downstream signaling protein of ERK1, ribosomal protein S6 kinase alpha 3 (RSK2) is activated through the "classical" MAPK pathway. So RSK2 plays a critical role in response to innate immune system induced by TRL3. However, the innate immune mechanism of RSK2 remains indistinct in fish. In this study, we cloned and characterized a full length cDNA sequence of RSK2 from Ctenopharyngodon idella (named CiRSK2, MH844551). The full length cDNA of CiRSK2 is 3930 bp with a coding sequence of 2202 bp encoding a polypeptide of 734 amino acids. The expression of CiRSK2 was ubiquitous and significantly up-regulated under the stimulation of poly (I:C) in eight different tissues of C. idella and C. idella kidney cells (CIK). In addition, poly (I:C) stimulation also up-regulated the expression of CiERK1 mRNA in CIK cells and the phosphorylation of CiERK1. We also demonstrated that the activated CiERK1 interacted with CiRSK2 by CO-IP assay and immunofluorescence assay. To further investigate the relationship between CiRSK2 and CiERK1, we performed subcellular localization of CiRSK2 at different periods of CiERK1 stimulation. The result showed that CiERK1 can make CiRSK2 enter the nucleus. Subsequently, we found that CiRSK2 increased the transcriptional level of CiBCL-2 and protein level of CiBCL-2 significantly. Then cell apoptosis was inhibited to a certain extent. Overall, our results suggested that CiRSK2 plays important roles in fish innate immunity and is able to inhibit cell apoptosis by up-regulating CiBCL-2.

Here, a pH-responsive ZnO nanoprobe mediated DNAzyme signal amplification strategy was proposed for sensitive detection and live cell imaging of multiple microRNAs. The nanoprobe including ZnO nanoparticles (ZnO NPs) core as carrier and cofactor provider and polydopamine shell for adsorption the functional hairpin DNAs was designed. When nanoprobe entered the cell through endocytosis, the acidic environment of the cell could decompose the ZnO NPs core to release the functional hairpin DNAs and Zn2+. The Zn2+ could act as cofactor for the DNAzyme cleavage amplification reaction, avoiding additional cell delivery processes. The recognition hairpin DNAs (H1 and H3) recognized microRNA and exposed the DNAzyme. The DNAzyme cleaved cyclically the reporter hairpin DNAs (H2 and H4), producing enhanced fluorescent signal for miR-21 and miR-373 detection with the detection limit of 54 pM and 38 pM, respectively. Expression levels of miR-21 in Hela, HepG-2 and L02 cells were differentiated. Furthermore, simultaneous imaging of miR-21 and miR-373 in same living cells was achieved. These results indicated this strategy could have a potential application in microRNAs assays for the accurate diagnosis and therapy of cancer.

MicroRNA (miRNA) plays significant roles in cell proliferation, differentiation and apoptosis, and has been considered to be valuable biomarker for cancer. Accurate and sensitive detection of miRNA is crucially significant for cancer diagnosis and treatment. Here, a MnO2 nanosheet-mediated ratiometric fluorescence biosensor was designed for miRNA detection and imaging in living cells. It contained MnO2 nanosheets acting as DNA carrier, and fluorescent donor (FAM)-labeled hairpin H1 (recognition probe) and fluorescent acceptor (TAMRA)-labeled hairpin H2 (amplification probe). When the biosensor entered cell by endocytosis, MnO2 nanosheets were degraded to Mn2+ via intracellular glutathione (GSH) and the adsorbed hairpins H1 and H2 were released. The intracellular target miRNA-21 hybridized with the recognition unit of H1 to initiate catalyzed hairpin assembly (CHA) and a large amount of H1-H2 duplexes were produced. This brought fluorescent donor FAM and fluorescent acceptor TAMRA into close proximity to produce fluorescence resonance energy transfer (FRET), inducing a ratiometric fluorescent response (donor signal decreased and acceptor signal enhanced) for miRNA-21 detection. Furthermore, this method could be applied to differentiate the expression levels of miRNA-21 in HeLa, HepG-2 and L02 cells. These results indicated that the proposed method possessed great potential in the early diagnosis of miRNA-related diseases.

Gesture recognition is important in human–machine interaction. Current methods for solving gesture recognition have several disadvantages such as low recognition rate, slow speed and poor performance on recognizing multiple targets or long-distance targets in complex environments. In view of the above problems, we propose a gesture recognition approach that can recognize gestures quickly and accurately from complex background. This approach works on a deep convolutional network, which consists of a basic network module for extracting feature information, a squeeze-and-excitation networks for increasing feature channel affinity and a feature pyramid attention module for fusing context information with different scales. To test the proposed approach, we make a data set that contains 3289 images from difference complex scenes. Generally gestures in those images can be generally classified into 16 types. We have uploaded this data set for researchers use. Experimental results demonstrate that the recognition accuracy and speed of the proposed method can achieve 83.45% and 32.2 frames per second respectively, which has better comprehensive performance compared with other state-of-the-art recognition algorithms.

Colletotrichum higginsianum causes anthracnose disease in a wide range of cruciferous crops and has been used as a model system to study plant-pathogen interactions and pathogenicity of hemibiotrophic plant pathogens. Conidiation, hyphae growth, appressorial development and appressorial penetration are significant steps during the infection process of C. higginsianum. However, the mechanisms of these important steps during infection remain incompletely understood. To further investigate the mechanisms of the plant-C. higginsianum interactions during infection progress, we characterized Cyclase-Associated Protein (ChCAP) gene. Deletion of the ChCAP gene resulted in reduction in conidiation and hyphal growth rate. The pathogenicity of ΔChCAP mutants was significantly reduced with much smaller lesion on the infected leaves compared to that of wild type strain with typically water-soaked and dark necrotic lesions on Arabidopsis leaves. Further study demonstrated that the appressorial formation rate, turgor pressure, penetration ability and switch from biotrophic to necrotrophic phases decreased obviously in ΔChCAP mutants, indicating that the attenuated pathogenicity of ΔChCAP mutants was due to these defective phenotypes. In addition, the ΔChCAP mutants sectored on PDA with abnormal, dark color, vesicle-like colony morphology and hyphae tip. Moreover, the ΔChCAP mutants had a reduced intracellular cAMP levels and exogenous cAMP can partially rescue the defects of ΔChCAP mutants in appressorial formation and penetration rate, but not in colony morphology, conidial shape and virulence, indicating that ChCAP is a key component in cAMP signaling pathway and likely play other roles in biology of C. higginsianum. In summary, our findings support the role of ChCAP in regulating conidiation, intracellular cAMP level, hyphal growth, appressorial formation, penetration ability and pathogenicity of this hemibiotrophic fungus.

Adenosine is closely related to the development of cancer, and it can be regarded as a biomarker for cancer diagnosis and therapy. Here, a T7 exonuclease (T7 Exo)-assisted and target-triggered cascade dual recycling signal amplification strategy was developed for the sensitive and specific detection of adenosine. In this strategy, a capture strand (Cap)-inhibit strand (Inh) duplex and a DNA hairpin with intact G-quadruplex sequence are designed. In the presence of adenosine, Cap binds with adenosine specifically to form Cap-adenosine complex with recessed 5′-hydroxyl termini, causing the release of Inh. Under the action of T7 Exo, the adenosine is released from Cap-adenosine complex. Then the released adenosine interacts with the next Cap-Inh duplex to promote the release of a new Inh and the recycle I is completed. After that, the released Inh hybridizes with hairpin to form Inh-hairpin duplex with blunt 5′ -hydroxyl termini. Under the same action of T7 Exo, the Inh gets released and a G-quadruplex sequence is obtained. Subsequently, the released Inh continues opening hairpin and the recycle II is accomplished. Finally, plenty of G-quadruplex sequences are generated and then interact with N-methyl-mesoporphyrin IX (NMM) to obtain enhanced fluorescence signal. The limit of detection (LOD) of the proposed strategy is estimated to be 9.8 × 10−7 mol L−1, and the linear range of the strategy is from 5.0 × 10−6 mol L−1 to 7.0 × 10−4 mol L−1. Besides, the proposed strategy is capable of distinguishing adenosine from its analogues. This strategy holds promise in adenosine related biomedical research, clinical diagnosis and disease treatment.

SAMHD1 is an innate immunity restriction factor that inhibits virus infection through IRF3-mediated antiviral and apoptotic responses. Fish SAMHD1 shares some similar properties with those in mammals. In this study, a SAMHD1 orthologue from grass carp (Ctenopharyngodon idellus) was cloned and characterized. The full-length cDNA of CiSAMHD1 is 2792 bp with an ORF of 1884 bp encoding a polypeptide of 627 amino acids. Multiple alignments showed that SAMHD1 is highly conserved among different species. Phylogenetic tree analysis revealed that CiSAMHD1 shared a high degree of homology with Sinocyclocheilus rhinocerous SAMHD1. Expression analysis indicated that CiSAMHD1 was widely expressed in all tissues tested including the brain, eyes, spleen, gill, intestine, liver, heart and kidney. It was significantly up-regulated in spleen, liver and intestines after treatment with poly I:C. Also, CiSAMHD1 can be induced following stimulation with recombinant IFN in CIK cells. The promoter sequence of CiSAMHD1 was identified to explore the mechanism underlying the transcriptional regulation of CiSAMHD1. The promoter sequence of CiSAMHD1 (1370 bp) consists of IRF1, IRF3, IRF9 and p65 binding elements. Gel mobility shift assay also showed that IRF1, IRF3, IRF9 and p65 prokaryotic proteins can separately interact with CiSAMHD1 promoter. Dual luciferase assay and q-PCR suggested that the promoter of CiSAMHD1 can be activated by the overexpression of CiIRF3 and CiIRF9, but cannot be triggered by CiIRF1 and Cip65. In contrast, knockdown of CiIRF3 or CiIRF9 inhibits the transcription of CiSAMHD1. Intriguingly, CCK assay suggested that CiSAMHD1 decreased cell viability. TUNEL apoptosis assay and Hoechst 33258 staining assay indicated that apoptosis is induced by the overexpression of CiSAMHD1. Crystal violet staining, detection of two GCRV genes (vp3 and vp5) and viral titration showed that CiSAMHD1 can suppress the proliferation of grass carp reovirus (GCRV) in CIK cells.

Two-dimensional three-temperature (2-D 3-T) energy equations are a type of strongly nonlinear system that describe the energy diffusion and exchange between an electron and a photon or ion. In multi-physics simulations, the process of energy diffusion and exchange is coupled with some other physical processes, such as fluid dynamics. Typically, the Lagrangian method is employed in radiation hydrodynamics simulations. Consequently, the 3-T energy equations should be discretized on deforming meshes, which are moved with dynamics. In 2-D deforming meshes, a nine-point scheme must be employed to discretize the 3-T energy equations. Because the energy diffusion and swapping coefficients are strongly nonlinearly dependent on the temperature, and some physical parameters are discontinuous across the material interfaces, it is a challenge to solve the discretized nonlinear algebraic equations in multi-physics and multi-material applications. A Newton-Krylov method is used to solve the discretized 2-D 3-T energy equations. Based on the physical properties and the characteristics of the equations, four types of operator-based preconditioners are constructed. Numerical results show that all the preconditioning methods considered in this study are very effective.

DNA methyltransferase (MTase) aberrant expression has a close relationship to tumorigenesis. DNA MTase activity detection is of great importance to its biomedical research and theranostics study. Here, multiple sealed primers-mediated rolling circle amplification (RCA) strategy is developed for sensitively and specifically detecting DNA MTase activity. The DNA probe has a folded, double-loop structure that seals multiple primers. First, in the presence of DNA MTase, the DNA probe is methylated, which then gets cleaved by the restriction endonuclease and breaks into multiple DNA oligonucleotide fragments. Second, each DNA oligonucleotide fragment acts as an independent primer for triggering RCA reaction respectively, producing long DNA strands that contain several interval G-quadruplexes. Finally, copious of G-quadruplexes are obtained, which bind N-methylmesoporphyrin IX (NMM) to generate significantly enhanced fluorescence. When DNA MTase is absent or inactive, the DNA probe is stable and cannot release the primers for RCA reaction. In the proposed strategy, the action of DNA MTase on one DNA probe is converted to the multiple amplifications triggered by multiple released primers. The detection limit for Dam MTase is down to 0.0085 U/mL, and the target MTase can be well discriminated from its MTases analogues. The method is utilized in screening of Dam MTase inhibitors and analyzing of spiked Dam MTase in biological samples. The results suggest that the strategy may provide a promising tool for DNA MTase activity detection in biomedical research and cancer theranostics.

Isoprocarb is a widely used carbamate insecticide in agriculture and aquaculture. Overuse of isoprocarb always leaves toxic residues in soil and water, however, the potential ecotoxicity of isoprocarb to organisms is still confusing. In this study, zebrafish embryo was used as a model to evaluate the toxicity of isoprocarb. Zebrafish embryos (96 hpf) were separately exposed at different concentrations of isoprocarb. The mortality rate, hatchability rate, average heart beat of the zebrafish embryo were separately calculated. Our results suggested that exposure to isoprocarb induced developmental toxicity in zebrafish embryos. HE staining showed that exposure to isoprocarb caused developmental defect in the hindbrain of zebrafish embryos. As expected, the behavioral analysis also showed that the motor ability of zebrafish embryos were significantly inhibited following exposure to isoprocarb. In terms of mechanism, The expressions of genes involved in neurodevelopment signaling pathways, such as foxo3a, gfap, syn2a, elavl3 and sox19b, were inhibited in zebrafish embryos after exposure to isoprocarb. The acetylcholinesterase (AChE) activity was also reduced in isoprocarb-treated zebrafish embryos. Moreover, oxidative stress was induced by increasing the reactive oxygen species (ROS) level and decreasing the activity of antioxidant enzyme (SOD) after exposure to isoprocarb. Expectedly, acridine orange (AO) staining and the detection of some apoptosis-related genes revealed that oxidative stress resulted in apoptosis. In short, the expressions of genes associated with the neurodevelopmental signaling pathway are inhibited, and oxidative stress is also induced in zebrafish embryos after exposure to isoprocarb, which may be the molecular basics of isoprocarb-induced neurotoxicity in zebrafish embryos.

PKR plays a significant role in IFN antiviral responses and in mediating apoptosis. Its activity is crucial for cellular antiviral and subsequent recovery. In mammalian cells, Protein Activator of the Interferon-induced Protein Kinase (PACT) and Trans-Activation-Responsive RNA-Binding Protein 2 (TARBP2) have the opposite effect on PKR activity in a dsRNA independent manner. There are some corresponding regulators of PKR in fish, too. In previous studies, we found that grass carp PACT can activate PKR in dsRNA independent manner. In this study, we tried to find out the effect of grass carp TARBP2 on PKR regulation. Grass carp TARBP2 expression is significantly increased at 6h post-poly I:C stimulation in CIK cells and grass carp tissues, indicating that it may play a role in poly I:C-mediated immune response. Then, we found that CiTARBP2 interacts with CiPKR and CiPACT, suggesting that it may regulate PKR activity by direct interaction with PKR or its regulators. Further, poly I:C promotes the phosphorylation of CiTARBP2 and enhances the interaction of CiTARBP2 and CiPKR. Finally, over-expression of CiTARBP2 decreases CiPKR phosphorylation and inhibits PKR-induced apoptosis. Therefore, our study reveals that CiTARBP2 can bind to CiPKR, CiPACT and CiTARBP2. The phosphorylated TARBP2 has stronger affinity to PKR, which results in the decrease of PKR phosphorylation and inhibition of cell apoptosis.

Herein, an enzyme-free and label-free fluorescence microRNA biosensor based on cascade amplification of DNAzyme-powered three-dimensional DNA walker and hybridization chain reaction was developed. Firstly, the microRNA hybridized with the locking DNA and caused the locking DNA released from the blocked walking probes (BWPs) through toehold-mediated strand displacement reaction. The DNAzyme was exposed and then the walking probes automatically moved along the three-dimensional tracks and catalyzed the cleavage of hairpin DNA substrates (HDSs), releasing abundant triggers. Finally, the triggers initiated the hybridization chain reaction to generate amplified fluorescence signals. Compared with the three-dimensional DNA walker powered by protein enzyme, this DNAzyme-powered three-dimensional DNA walker was simpler and more stable. The high amplification efficiency of cascade amplification guaranteed a satisfactory sensitivity with a detection limit of 7.9 fM. The toehold-mediated strand displacement reaction guaranteed an admirable specificity. The microRNA analysis in real biological sample further indicated that this biosensor offered an alternative strategy for the quantification of microRNA.

Objective To determine the clinical features of confirmed cases of severe fever with thrombocytopenia syndrome (SFTS) and to explore the early warning indicators of death from SFTS. Methods A retrospective case–control study was performed at a single medical institution in Yantai. A total of 20 SFTS patients who died (death group) during January 2014 to December 2015 and another 40 age- and sex-matched SFTS patients who survived (survivor group) were identified from the case records. The differences in demographic characteristics, clinical signs and symptoms, and laboratory parameters in the early stage of disease were compared between the two groups. Conditional logistic regression was used to identify the independent risk factors for mortality in SFTS patients. Results Univariate logistic regression analysis showed that a disturbance of consciousness, pulse–temperature deficit, neurological signs, hemorrhagic manifestations, pulmonary infection, decreased lymphocyte percentage, high lactate dehydrogenase and creatine kinase levels, increased serum creatinine, blood urea nitrogen, and C-reactive protein (CRP), hyponatremia, and prolonged activated partial thromboplastin time (APPT) and prothrombin time were associated with mortality. On multivariate logistic regression analysis, the independent predictors of death were neurological signs (odds ratio (OR) 31.247, 95% confidence interval (CI) 4.813–202.853), hemorrhagic manifestations (OR 20.251, 95% CI 2.056–199.443), disturbance of consciousness (OR 15.359, 95% CI 2.139–110.268), hyponatremia (OR 5.280, 95% CI 1.235–22.575), increased CRP (OR 2.641, 95% CI 1.090–6.396), increased serum creatinine (OR 6.776, 95% CI 1.047–43.840), and prolonged APTT (OR 6.018, 95% CI 1.450–24.975). Conclusions Neurological signs, hemorrhagic manifestations, disturbance of consciousness, hyponatremia, prolonged APTT, and increased CRP and serum creatinine are risk factors for death in SFTS.

Herein, an integrated and restructive probe mediated strand displacement amplification (SDA) strategy is developed for sensitive and specific DNA MTase activity detection. The probe is specially designed with a hairpin structure, which encloses the MTase recognition site in the stem and seals the SDA primer and template in the loop. Under the action of DNA MTase, the probe is methylated and cleaved by DpnI endonuclease, causing its stem truncated. The truncated structure then reconstructs with a shrunken loop and a partially-hybridized duplex of stem, drawing the SDA primer and template domains close to trigger cascade amplification. Numerous G-quadruplexes are produced and intercalated by N-methyl-mesoporphyrin IX (NMM) to generate enhanced fluorescent signal. This strategy detects MTase activity down to 0.063 U/mL, and well distinguishes Dam MTase from its analogues. It is further applied for the MTase detection in biological samples and MTase inhibition analysis. The strategy will provide a promising tool for detection MTase activity in biomedical study and early cancer diagnosis.

Human telomerase is an endogenous ribonucleoprotein that is over-expressed in most types of malignant cancer cells. Sensitive and specific detection of telomerase activity is crucial for better understanding its role in cancer cells and further exploring its function in cancer diagnosis. Here, we develop convertible DNA ends-based silver nanoprobes for sensitive and specific colorimetric detection telomerase activity. Silver nanoprobes are constructed by modifying telomerase binding substrates (TS) that are pre-hybridized with complementary sequences onto silver nanoparticles (AgNPs), via the coordination between consecutive cytosines in TS strand and AgNPs. This forms blunt-end terminated, double-stranded DNA on the surface of AgNPs. Under the action of telomerase, TS on the silver nanoprobes are elongated with telomeric repeats, converting DNA stiff blunt ends to flexible single-stranded dangling ends. The dangling ends enhance the stability of nanoprobes and relieve their salt-induced aggregation, and the solution shows a yellow color. When telomerase is inactive, the blunt end-terminated nanoprobes cannot resist salt-induced aggregation, resulting in a gray color of solution. Based on telomerase-regulated DNA “blunt-dangling” ends conversion-induced AgNPs' dispersity and color change, colorimetric detection of the endogenous telomerase with AgNPs is realized. The detection limit is equivalent to 1 cell/μL of telomerase activity, and extracts from cancer cells and normal cells are visually distinguished through color difference. The proposed strategy will offer a new approach for reliable, convenient quantification of telomerase activity in biochemical research and clinical diagnosis.

DNA as a novel biomaterial can be used to fabricate different kinds of DNA nanostructures based on its principle of GC/AT complementary base pairing. Studies have shown that DNA nanostructure is a nice drug carrier to overcome big obstacles existing in cancer therapy such as systemic toxicity and unsatisfied drug efficacy. Thus, different types of DNA nanostructure-based drug delivery nanosystems have been designed in cancer therapy. To improve treating efficacy, they are also developed into more functional drug delivery nanosystems. In recent years, some important progresses have been made. The objective of this review is to make a retrospect and summary about these different kinds of DNA nanostructure-based drug delivery nanosystems and their latest progresses: (1) active targeting; (2) mutidrug co-delivery; (3) construction of stimuli-responsive/intelligent nanosystems.

DNA methyltransferase (MTase) is a predictive cancer biomarker and drug target, and the sensitive detection of DNA MTase activity is crucial to early cancer diagnosis and therapy. In this work, we developed a loop-mediated cascade amplification strategy for highly sensitive fluorescent detection of DNA MTase activity based on strand displacement amplification (SDA) and exponential rolling circle amplification (ERCA). Firstly, we designed a long stem-loop probe (LSLP) which contains a methylation site for DNA MTase recognition, a long stem for ensuring the stability of probe, and a loop for initiating subsequent amplification. The loop and part of stem of LSLP acted as a trigger strand for subsequent signal output process. And the trigger strand was fully enclosed in loop of LSLP by the long stem, avoiding the nonspecific amplification caused by the leakage of probe. The LSLP was methylated by DNA MTase and then was specifically cleaved by DpnI endonuclease, producing a trigger strand. Under the synergetic action of polymerase and nicking enzyme, the trigger strand initiated SDA, producing many primers. The produced primers initiated ERCA, synthesizing numerous G-quadruplex sequences. The G-quadruplex sequences interacted with N-methylmesoporphyrin IX, obtaining an enhanced fluorescent signal. The method could detect as low as 8.1 × 10−5 U/mL DNA MTase. Furthermore, this assay was successfully used to assess the inhibition effect of inhibitors for DNA MTase activity. These results show that our system has a great potential in early cancer diagnosis and therapy.

PRKRA (interferon-inducible double-stranded RNA-dependent protein kinase activator A) is a protective protein which regulates the adaptation of cells to ER stress and virus-stimulated signaling pathways by activating PKR. In the present study, a grass carp (Ctenopharyngodon idella) PRKRA full-length cDNA (named CiPRKRA, KT891991) was cloned and identified. The full-length cDNA is comprised of a 5′ UTR (36 bp), a 3′ UTR (350 bp) and the longest ORF (882 bp) encoding a polypeptide of 293 amino acids. The deduced amino acid sequence of CiPRKRA contains three typical dsRNA binding motifs (dsRBM). Phylogenetic tree analysis revealed a closer evolutionary relationship of CiPRKRA with other fish PRKRA, especially with Danio rerio PRKRA. qRT-PCR showed that CiPRKRA was significantly up-regulated after stimulation with tunicamycin (Tm) and Poly I:C in C. idella kidney (CIK) cells. To further study its transcriptional regulation, the partial promoter sequence of CiPRKRA (1463 bp) containing one ISRE and one CARE was cloned by Tail-PCR. Subsequently, grass carp IRF2 (CiIRF2) and ATF4 (CiATF4) were expressed in Escherichia coli BL21 and purified by affinity chromatography with the Ni-NTA His-Bind Resin. In vitro, both CiIRF2 and CiATF4 bound to CiPRKRA promoter with high affinity by gel mobility shift assays, revealing that IRF2 and ATF4 might be potential transcriptional regulatory factors for CiPRKRA. Dual-luciferase reporter assays were applied to further investigate the transcriptional regulation of CiPRKRA in vivo. Recombinant plasmid of pGL3-PRKRAPro was constructed and transiently co-transfected into CIK cells with pcDNA3.1-CiIRF2 and pcDNA3.1-CiATF4, respectively. The results showed that both CiIRF2 and CiATF4 significantly decreased the luciferase activity of pGL3-PRKRAPro, suggesting that they play a negative role in CiPRKRA transcription.

The coefficient matrices of the linear equation systems arising from the radiation diffusion problems usually have orders of magnitude difference between their off-diagonal entries. While solving these linear equations with a preconditioned iterative method, the entries with small magnitudes may be insignificant to the preconditioner efficiency. In this paper, we use a filter to remove such small entries in the coefficient matrix while constructing the preconditioner. The proposed filter eliminates the small entries first according to a so-called weak dependence matrix, which relies on the conception of the strength of connections in algebraic multigrid. The preconditioner is then built based on the filtered matrix instead of the original one. Four strategies of filtering out entries are designed and investigated. Numerical results for various model-type problems and two real application problems, i.e., the multi-group radiation diffusion equations and the three temperature energy equations, are provided to show the effectiveness of the proposed method. In particular, this paper provides a practical approach to choose a proper parameter in the proposed method, which should help solve linear equation systems of radiation diffusion problems.

A novel MNAzyme signal amplification strategy was developed for enzyme-free and label-free detection of DNA-binding proteins. This strategy relied on the binding-mediated MNAzyme cleavage and G-quadruplex-based light-up fluorescence switch. Three DNA sequences were designed to construct the MNAzyme in which DNA1 (including half binding site of the target protein and a toehold sequence) and DNA2 (including another half binding site of the target protein and one MNAzyme partzyme) firstly hybridized. The target protein recognized the binding sites on DNA1-DNA2 hybrid to form a stable protein-DNA1-DNA2 conjugates. Then, the MNAzyme was assembled with the presence of DNA3 which contained another MNAzyme partzyme and the complementary sequence of DNA1. The active MNAzyme cleaved DNA4 to release the G-quadruplex that was locked in the stem of DNA4. Finally, N-methyl mesoporphyrin IX (NMM) was inserted into the released G-quadruplex structure and the fluorescence signal was turned on. Taking nuclear factor-κB p50 (NF-κB p50) as the model, the limit of detection was low to 0.14 nM. Furthermore, the sequence-specific recognition of NF-κB p50 with DNA displayed excellent selectivity and specificity. The results in present work showed that this strategy will be a promising tool for DNA-binding proteins analysis in biomedical exploration and clinical diagnosis.

In the regulatory network, miRNAs play a regulatory role in a cooperative or antagonistic manner. Simultaneous accurate detection and imaging of multiplexed miRNAs in living cells are of great significance for miRNA-associated biological research and disease diagnosis and treatment. Herein, a MnO2 nanosheet-mediated target-binding-induced fluorescence resonance energy transfer (FRET) strategy was developed for detection and imaging of multiplexed miRNAs in living cells. Two pairs of DNA probes (P1-AF 488/P1′-Cy3 and P2-AF 488/P2′-AF 594) contained the complementary sequence to target miRNAs (miRNA-373 and miRNA-96) and labelled with different fluorescence dyes were designed. They were adsorbed onto MnO2 nanosheets by physisorption to form DNA/MnO2 nanocomposite probes. When the DNA/MnO2 nanocomposite probes were taken up by cells, the MnO2 nanosheets were reduced by intracellular glutathione, accompanying the release of DNA probe pairs. Then the DNA probe pairs specifically recognized and combined with miRNA-373 and miRNA-96 to form stable duplexes, respectively, bringing labelled fluorophores into close proximity to occur FRET. Based on this, the simultaneous imaging of miRNA-373 and miRNA-96 in MDA-MB-231 and L02 cells was successfully implemented. The results displayed a higher expression level of target miRNAs in MDA-MB-231 cells compared to L02 cells. The changes in expression levels of miRNA-96 induced by anti-miRNA-96 or mimics in MDA-MB-231 cells could also be monitored. In addition, the ratiometric detections of multiplexed miRNAs were achieved by utilizing the DNA probe pairs. The proposed strategy provides an alternative method for simultaneous accurate detection and imaging of multiplexed miRNAs and has potential application in biomedical applications.

A green, novel and efficacious method for the simultaneous extraction and enrichment of rutin and narcissoside from the compound Anoectochilus roxburghii (Wall.) Lindl. oral liquid (CAROL) and Anoectochilus roxburghii (Wall.) Lindl. was developed. Ionic liquid-surfactant synergic cloud-point extraction (IL-CPE) was used to enrich two analytes, which were determined by high-performance liquid chromatography (HPLC). Some parameters affecting IL-CPE were optimized, such as ratio and volume of 1-butyl-3-methyl-imidazolium hexafluorophosphate and Triton X-114, pH of the sample, NaCl concentration, total extraction volume, incubation temperature and time, centrifuge rate and time. The corresponding linearity range for two analytes exhibited good linearity (r2＞0.9997), with the average added recoveries ranging from 92.1% to 98.9%. The limits of detection of rutin and narcissoside were 0.26 and 0.30 ng/mL, respectively. The method was successfully applied for the determination of two flavonoids in the complex-matrix sample, i.e. CAROL and the water extract of A. roxburghii. The mass spectrum data showed that the sample contained rutin and narcissoside. Compared with conventional extraction methods, IL-CPE exhibited higher extraction efficiency and better extraction selectivity. This method may provide a novel platform for the determination of active ingredients in compound Chinese medicine oral liquid and herb.

Here, a MNAzyme probes mediated DNA logic platform was developed for microRNAs (miRNAs) logic detection and cancer cells identification. A series of MNAzyme probes containing the cleavage active center were designed. Four types of logic gates were constructed, including YES, AND, XOR and NOR gate. These logic gates used miRNAs that were high expression in cancer cells as logic inputs and used MNAzyme cleavage amplification reaction to output signals. For the construction of intracellular logic gates, MnO2 nanosheets were used as carriers and cofactor providers. When MnO2 nanoprobes entered the cells through endocytosis, the intracellular glutathione degraded the MnO2 nanosheets to release the cofactor Mn2+ and MNAzyme probes. The MNAzyme probes bound to the miRNAs and catalyze the MNAzyme cleavage amplification reaction, producing enhanced fluorescent signal with "true" output. The logic detection of miRNAs was achieved by integrating information from the AND, XOR and NOR logic gates. Moreover, through the construction of intracellular YES and AND logic gates, the cancer cells identification, especially the identification of same type of cancer cells with different phenotypes was achieved. These experimental results showed that this platform held great promise in accurate diagnosis and treatment of cancer.

Herein, a bipedal-unequivalent three-dimensional DNA walker is designed and applied to biosensing. It involves three components: (i) DNA three-way junction switch (DTWJS), in which two longer strands containing different sequences are partially complementary serving as bipedal-unequivalent walking strand (B-UWS) and the shortest one serves as blocking strand; (ii) fluorophore-labelled hairpins H1 and H2, co-modified onto gold nanoparticles (AuNPs), serving as track strands; (iii) hairpins H3 and H4, contain sequences complementary to H1 and H2, respectively, serving as fuel strands. By target binding to blocking strand, B-UWS is liberated to hybridize with H1 and H2 in turn. The unfolded H1 and H2 get hybridized with H3 and H4, respectively, accompanying by B-UWS sequential release and fluorescence signal accumulation. Then B-UWS continues to hybridize with another track strands to walk. Compared with unipedal and bipedal-equivalent DNA walkers, this bipedal-unequivalent DNA walker has twice the sustainable operation capability through kinetic and affinity studies. Using let-7a as a model target, it shows a detection limit of 68 pM and satisfactory reproducibility in biological fluids. This DNA walker provides a new sustainable operation mode and will be a potential analytical tool in clinical diagnosis.

As one member of ADAR family, ADAR1 (adenosine deaminase acting on RNA 1) can convert adenosine to inosine within dsRNA. There are many ADAR1 splicing isoforms in mammals, including an interferon (IFN) inducible ∼150 kD protein (ADAR1-p150) and a constitutively expressed ∼110 kD protein (ADAR1-p110). The structural diversity of ADAR1 splicing isoforms may reflect their multiple functions. ADAR1 splicing isoforms were also found in fish. In our previous study, we have cloned and identified two different grass carp ADAR1 splicing isoforms, i.e. CiADAR1 and CiADAR1-like, both of them are IFN-inducible proteins. In this paper, we identified a novel CiADAR1 splicing isoform gene (named CiADAR1a). CiADAR1a gene contains 15 exons and 14 introns. Its full-length cDNA is comprised of a 5′ UTR (359 bp), a 3′ UTR (229 bp) and a 2952 bp ORF encoding a polypeptide of 983 amino acids with one Z-DNA binding domain, three dsRNA binding motifs and a highly conserved hydrolytic deamination domain. CiADAR1a was constitutively expressed in Ctenopharyngodon idella kidney (CIK) cells regardless of Poly I:C stimulation by Western blot assay. In normal condition, CiADAR1a was found to be present mainly in the nucleus. After treatment with Poly I:C, it gradually shifted to cytoplasm. To further investigate the mechanism of transcriptional regulation of CiADAR1a, we cloned and identified its promoter sequence. The transcriptional start site of CiADAR1a is mapped within the truncated exon 2. CiADAR1a promoter is 1303 bp in length containing 4 IRF-Es. In the present study, we constructed pcDNA3.1 eukaryotic expression vectors with IRF1 and IRF3 and co-transfected them with pGL3-CiADAR1a promoter into CIK cells. The results showed that neither the over-expression of IRF1 or IRF3 nor Poly I:C stimulation significantly impacted CiADAR1a promoter activity in CIK cells. Together, according to the molecular and expression characteristics, subcellular localization and transcriptional regulatory mechanism, we deduced that CiADAR1a shared a high degree of homology with mammalian ADAR1-p110.

NF-κB is an important transcription factor for regulating the multiple inflammatory and immune related gene transcription. It can bind with the nuclear factor κB site within the promoter of target genes to regulate their transcriptions. p65, the all-important subunit of NF-κB, is ubiquitously expressed in cells. In the present study, we cloned and identified the p65 subunit from grass carp (Ctenopharyngodon idella) (named Cip65) by homologous cloning and RACE technique. The full length of Cip65 cDNA is 2481 bp along with 9 bp 5' UTR, 639 bp 3' UTR and the largest open reading frame (1833 bp) encoding a polypeptide of 610 amino acids with a well conserved Rel-homology domain (RHD) in N-terminal and a putative transcription activation domain (TAD) in C-terminal. Cip65 gathers with other teleost p65 proteins to form a fish-specific clade clearly distinct from those of mammalian and amphibian counterparts on the phylogenetic tree. In CIK (C. idellus kidney) cells, the expression of Cip65 was significantly up-regulated under the stimulation with Poly I:C. As one member of the NF-κB inhibitor protein (IκB) family, IκBα can dominate the activity of NF-κB by interacting with it. To study the molecular mechanisms of negative feedback loop of NF-κB signaling in fish, we cloned grass carp IκBα (CiIκBα) promoter sequence. CiIκBα promoter is 414 bp in length containing two RelA binding sites and a putative atypical TATA-box. Meanwhile, Cip65 and its mutant proteins including C-terminus deletion mutant of Cip65 (Cip65-ΔC) and N-terminus deletion mutant of Cip65 (Cip65-ΔN) were expressed in Escherichia coli BL21 and purified by affinity chromatography with the Ni-NTA His-Bind resin. In vitro, Cip65 rather than Cip65-ΔC and Cip65-ΔN showed high affinity with CiIκBα promoter sequence by gel mobility shift assays. In vivo, the cotransfection of pcDNA3.1-Cip65 (or pcDNA3.1-Cip65-ΔC, pcDNA3.1-Cip65-ΔN respectively) with pGL3-CiIκBα and pRL-TK renilla luciferase plasmid into CIK cells showed that pcDNA3.1-Cip65 rather than pcDNA3.1-Cip65-ΔC and pcDNA3.1-Cip65-ΔN, can increase the luciferase activity. Taken together, these results suggested that Cip65 can regulate the expression of CiIκBα and works as a negative feedback loop in NF-κB pathway.

Protein kinase R (PKR), the double-stranded RNA-activated protein kinase, exists in mammalian and fish. PKZ, a PKR-like protein kinase containing Z-DNA binding domains, just exists in fish. PKR and PKZ work synergistically in the antiviral defense by inhibiting intracellular protein translation. The transcriptional factor IRF3 (interferon regulatory factor 3) acts as a key regulator of type I IFN (Interferon) and ISG (interferon stimulated gene). On the basis of the cloned CiIRF3 previously, CiIRF3 with His-tag was over-expressed in BL21 Escherichia coli, and the expressed protein was purified by affinity chromatography with Ni-NTA His-Bind Resin. In this study, we have demonstrated that grass carp (Ctenopharyngodon idellus) PKR (CiPKR) and PKZ (CiPKZ) genes were inducible by Poly I:C in C. idella kidney (CIK) cells. So, they might be implicated in the intracellular antiviral activity. To understand the up regulatory mechanism of CiPKR and CiPKZ genes upon virus induction, we constructed wild type (pGL3-CiPKR-luc and pGL3-CiPKZ-luc) and the mutant (pGL3-CiPKR-nISRE-luc and pGL3-CiPKZ-nISRE-luc) reporter gene vectors according to the promoter sequences of CiPKR (KJ704845) and CiPKZ (KJ704844). In vitro, gel mobility shift assays demonstrated that CiIRF3 can combine CiPKR and CiPKZ promoters with high affinity. However, CiIRF3 bound to the mutants CiPKR-nISRE and CiPKZ-nISRE faintly. Whereafter, the recombinant plasmids of pGL3-CiPKR-luc, pGL3-CiPKZ-luc were transiently co-transfected with pcDNA3.1-CiIRF3, pcDNA3.1-CiIRF7 respectively into CIK cells. Cell transfection assays indicated that CiIRF3 and CiIRF7 up-regulated the transcriptional level of CiPKR and CiPKZ. The results also revealed that the consensus sequence of ISRE (interferon stimulated response element) is an important regulatory element for the transcriptional initiation of CiPKR and CiPKZ.

GPATCH3, a protein with G-patch domain, is known to participate in innate immune response and organ development in mammals. However, there are few reports on GPATCH3 in fish. Here the cDNA sequence of GPATCH3 was cloned from Ctenopharyngodon idella (CiGPATCH3, MN149902) and was determined its character. A cDNA sequence of CiGPATCH3 is 1646 bp and contains an ORF of 1221 bp translating a protein of 407 amino acids. Phylogenetic analysis uncovered that CiGPATCH3 possesses a relatively high degree of homology with Cyprinus carpio GPATCH3. The mRNA level of CiGPATCH3 was increased following the intracellular stimulation of poly (I:C) into CIK cells. In vivo, over-expression of CiGPATCH3 can significantly up-regulate IFN 1 and ISG15 expression at mRNA and protein levels. To investigate the molecular mechanism by which GPATCH3 initiates the innate immune response in fish, co-IP experiments were performed to analyze the substrates of CiGPATCH3. The results showed that CiGPATCH3 directly interacted with CiSTING, but not with CiIRF3, CiIRF7, CiTBK1 or CiIPS-1. As compared with the single transfection of CO cells with either CiGPATCH3 or CiSTING, the expression of IFN 1 was more significantly up-regulated in cells under treatment with dual transfection of CiGPATCH3 and CiSTING. Knockdown of CiGPATCH3 inhibited STING-mediated IFN 1 expression in fish cells. Over-expression of CiGPATCH3 and CiSTING facilitated the phosphorylation and cytoplasmic-to-nuclear translocation of CiIRF7. These results explicitly showed that CiGPATCH3 up-regulates IFN 1 and ISG15 expression via the activation of STING-IRF7 signal axis in vivo.

Herein, a track-regenerated DNA walker is constructed and applied for sensitive detection. Blocker-silenced walking strands and linear DNAs serving as roadbed strands are co-modified onto gold nanoparticles. Fluorophore/quencher dually-labeled hairpin DNAs serving as rail strands are free in solution. Roadbed strands hybridize with rail strands to form the track. Activated by the stimulus specifically binding to blocker, the walking strand will be liberated and hybridize with the rail strand to convert its 3′-end from protruding to recessed state. The rail strand then gets digested by exonuclease III, accompanying by fluorophore release and fluorescence enhancement. The vacant roadbed strand anew binds free rail strands, resulting in the regeneration of track. The walking strand continues to interact with the next rail strand and constantly steps forward on the regenerative track, accumulating large amounts of signal. This feature allows the DNA walker to be tailored into a sensor. For Ebola virus detection, it obtains a detection limit down to 3.5 fM and detects the target in biological fluids with good recovery. The DNA walker offers a new mode of sustainable operation and will provide a sensitively and reliably analytical tool in diagnosis.

The present study investigated the neural responses to errors in a shooting game - and how these neural responses may relate to behavioral performance - by examining the ERP components related to error detection (error-related negativity; ERN) and error awareness (error-related positivity; Pe). The participants completed a Shooter go/no-go task, which required them to shoot at armed targets using a gaming gun, and avoid shooting innocent non-targets. The amplitude of the ERN and Pe was greater for shooting errors than correct shooting responses. The ERN and Pe amplitudes elicited by incorrect shooting appeared to have good internal reliability. The ERN and Pe amplitudes elicited by shooting behaviors also predicted better behavioral sensitivity towards shoot/don't-shoot stimuli. These results suggest that it is possible to obtain online brain response measures to shooting responses and that neural responses to shooting are predictive of behavioral responses.

This paper describes the development and validation of a liquid chromatography–mass spectrometric assay for propafenone and its application to a pharmacokinetic study of propafenone administered as a new propafenone hydrochloride sustained-release capsule (SR-test), as an instant-release tablet (IR-reference) and as the market leader sustained-release capsule (Rythmol, SR-reference) in male beagle dogs (n=8). In Study A comparing SR-test with IR-reference in a crossover design Tmax and t1/2 of propafenone for SR-test were significantly higher than those for IR-reference while Cmax and AUC were lower demonstrating the sustained release properties of the new formulation. In Study B comparing SR-test with SR-reference the observed Cmax and AUC of propafenone for SR-test (124.5±140.0 ng/mL and 612.0±699.2 ng·h/mL, respectively) were higher than for SR-reference (78.52±72.92 ng/mL and 423.6±431.6 ng·h/mL, respectively) although the differences were not significant. Overall, the new formulation has as good if not better sustained release characteristics to the market leader formulation., Graphical abstract This paper compares the pharmacokinetics of propafenone administered as a new formulation of sustained-release capsules with those for instant-release tablets and the market leader sustained-release capsules in male beagle dogs. The results show the new sustained-release capsule provides significantly slower release than from the instant-release tablet and similar pharmacokinetics to the market leader sustained-release capsules.

In this paper, we consider the properties of monotonicity-preserving and global conservation- preserving for interpolation operators. These two properties play important role when interpolation operators used in many real numerical simulations. In order to attain these two aspects, we propose a one-dimensional (1D) new cubic spline, and extend it to two-dimensional (2D) using tensor-product operation. Based on discrete convolution, 1D and 2D quasi-interpolation operators are presented using these functions. Both analysis and numerical results show that the interpolation operators constructed in this paper are monotonic and conservative. In particular, we consider the numerical simulations of 2D Euler equations based on the technique of structured adaptive mesh refinement (SAMR). In SAMR simulations, effective interpolators are needed for information transportation between the coarser/finer meshes. We applied the 2D quasi-interpolation operator to this environment, and the simulation result show the efficiency and correctness of our interpolator. (C) 2010 Elsevier Inc. All rights reserved.

Bismuth molybdate (Bi2MoO6) nanoparticles with the size of 10 nm were prepared on NaY type zeolite by the impregnation with Bi(NO3)3 and (NH4)6Mo7O24 solution and subsequent calcination at different temperatures. The mechanism of the formation of Bi2MoO6 nanoparticles on NaY type zeolite is discussed based on the adsorption data at various temperatures. The photocatalytic activities of the Bi2MoO6/NaY (BM) were evaluated by the degradation of methyl orange (MO) in aqueous solution under visible light irradiation.

The 2-D 3-T heat conduction equations can be used to approximately describe the energy broadcast in materials and the energy swapping between electron and photon or ion. To solve the equations, a fully implicit finite volume scheme is often used as the discretization method. Because the energy diffusion and swapping coefficients have a strongly nonlinear dependence on the temperature, and some physical parameters are discontinuous across the interfaces between the materials, it is a challenge to solve the discretized nonlinear algebraic equations. Particularly, as time advances, the temperature varies so greatly in the front of energy that it is difficult to choose an effective initial iterate when the nonlinear algebraic equations are solved by an iterative method. In this paper, a method of choosing a nonlinear initial iterate is proposed for iterative solving this kind of nonlinear algebraic equations. Numerical results show the proposed initial iterate can improve the computational efficiency, and also the convergence behavior of the nonlinear iteration.