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Soybean is a short-day plant that typically flowers earlier when exposed to short-day conditions. However, the identification of genes associated with earlier flowering time but without a yield penalty is rare. In this study, we conducted genome-wide association studies (GWAS) using two re-sequencing datasets that included 113 wild soybeans (G. soja) and 1192 cultivated soybeans (G. max), respectively, and simultaneously identified a candidate flowering gene, qFT13-3, which encodes a protein homologous to the pseudo-response regulator (PRR) transcription factor. We identified four major haplotypes of qFT13-3 in the natural population, with haplotype H4 (qFT13-3 H4 ) being lost during domestication, while qFT13-3 H1 underwent natural and artificial selection, increasing in proportion from 4.5% in G. soja to 43.8% in landrace and to 81.9% in improve cultivars. Notably, most cultivars harbouring qFT13-3 H1 were located in high-latitude regions. Knockout of qFT13-3 accelerated flowering and maturity time under long-day conditions, indicating that qFT13-3 functions as a flowering inhibitor. Our results also showed that qFT13-3 directly downregulates the expression of GmELF3b-2 which is a component of the circadian clock evening complex. Field trials revealed that the qft13-3 mutants shorten the maturity period by 11 days without a concomitant penalty on yield. Collectively, qFT13-3 can be utilized for the breeding of high-yield cultivars with a short maturity time suitable for high latitudes., (© 2023 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

Using electrochemical water splitting to generate hydrogen is considered a desirable approach, which is greatly impeded by the sluggish dissociation of H 2 O and adsorption and desorption of H*. Effective hydrogen production can be achieved by speeding up the chemical process with a suitable electrocatalyst. In this work, we designed and synthesized a rare earth element cerium (Ce) regulated iron-nickel bimetallic phosphide Ce-NiFeP@NF (here NiFeP represents Fe 2 P/NiP 2 ) nanoarray with nanoflowers. For the hydrogen evolution reaction (HER), Ce-NiFeP@NF only needs an overpotential of 106 mV to provide a current density of 10 mA cm -2 , compared to NiFeP@NF (175 mV@10 mA cm -2 ). This self-supported electrocatalyst Ce-NiFeP@NF with a composite morphology exhibits excellent performance in the HER. Specifically, the introduction of Ce promotes the electron transfer process at the Fe 2 P/NiP 2 heterojunction interface and the Ce-NiFeP@NF nanocomposite structure with nanoflowers has a larger electrochemically active specific surface area, which is more conducive to improving the intrinsic catalytic activity. Also, a dual-electrode alkaline electrolytic cell (Ce-NiFeP@NF functions as both the anode and the cathode) operates with a cell voltage of only 1.56 V to achieve a current density of 10 mA cm -2 . The synergistic effect of rare earth element doping and heterojunction engineering can improve the morphology of intrinsic catalysts to achieve more efficient electrochemical water splitting for hydrogen production.

The habenula (Hb) is involved in many natural human behaviors, and the relevance of its alterations in size and neural activity to several psychiatric disorders and addictive behaviors has been presumed and investigated in recent years using magnetic resonance imaging (MRI). Although the Hb is small, an increasing number of studies have overcome the difficulties in MRI. Conventional structural-based imaging also has great defects in observing the Hb contrast with adjacent structures. In addition, more and more attention should be paid to the Hb's functional, structural, and quantitative imaging studies. Several advanced MRI methods have recently been employed in clinical studies to explore the Hb and its involvement in psychiatric diseases. This review summarizes the anatomy and function of the human Hb; moreover, it focuses on exploring the human Hb with noninvasive MRI approaches, highlighting strategies to overcome the poor contrast with adjacent structures and the need for multiparametric MRI to develop imaging markers for diagnosis and treatment follow-up. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY STAGE: 2., (© 2023 The Authors. Journal of Magnetic Resonance Imaging published by Wiley Periodicals LLC on behalf of International Society for Magnetic Resonance in Medicine.)

Development of efficient and robust cathode catalysts is critical for the commercialization of Li-O 2 batteries (LOBs). Herein, a well-designed CePO 4 @N-P-CNSs cathode catalyst for LOBs via coupling P-N site-rich N, P co-doped graphene-like carbon nanosheets (N-P-CNSs) with nano-CePO 4 via a novel "in situ derivation" coupling strategy by in situ transforming the P atoms of P-C sites in N-P-CNSs to CePO 4 is reported. The CePO 4 @N-P-CNSs exhibit superior bifunctional ORR/OER activity relative to commercial Pt/C-RuO 2 with an overall overpotential of 0.64 V (vs RHE). Moreover, the LOB with CePO 4 @N-P-CNSs as the cathode catalyst delivers a low charge overpotential of 0.67 V (vs Li/Li + ), high discharge capacity of 29774 mAh g -1 at 100 mA g -1 and long cycling stability of 415 cycles, respectively. The remarkably enhanced LOB performance is attributable to the in situ derived CePO 4 nanoparticles and the P-N sites in N-P-CNSs, which facilitate increased bifunctional ORR/OER activity, promote the rapid and effective decomposition of Li 2 O 2 and inhibit the formation of Li 2 CO 3 . This work may provide new inspiration for designing efficient, durable, and cost-effective cathode catalysts for LOBs., (© 2023 Wiley-VCH GmbH.)

The development of biomass-based composites has greatly reduced the daily consumption of plastics. However, these materials are rarely recyclable, thus, posing a severe threat to the environment. Herein, we designed and prepared novel composite materials with ultra-high biomass (i.e., wood flour) filling capacity and good closed-loop recycling properties. The dynamic polyurethane polymer was polymerized in situ on the surface of wood fiber, and then they were hot-pressed into composites. Fourier-transform infrared (FTIR), scanning electron microscopy (SEM), and dynamic thermomechanical analysis (DMA) measurements reveal good compatibility between the polyurethane and wood flour in the composites when the wood flour content is ≤80 wt%. The maximum tensile and bending strength of the composite are 37 and 33 MPa when the wood flour content is 80%. The higher wood flour content results in higher thermal expansion stability and creep resistance in the composites. Moreover, the thermal debonding of dynamic phenol-carbamate bonds facilitates the composites to undergo physical and chemical cycling. The recycled and remolded composites exhibit good mechanical property recovery rates and retain the chemical structures of the original composites.

Objectives: Despite a high risk of human papillomavirus (HPV) infection among men who have sex with men (MSM), few have ever tested. This study aimed to evaluate the feasibility and accuracy of HPV self-sampling among Chinese MSM, with the purpose of measuring the feasibility of self-sampling as an alternative in HPV testing scenarios., Methods: Eligible participants were those who were assigned male at birth, aged 18 or above, had sex with men in the past year and had never gotten HPV vaccine. Participants followed the instructions to self-sample and were also clinician-sampled from the same anatomical sites (oral fluid, penis and rectum) in both approaches. All specimens were processed using multiplex PCR assay. The reference standard of an individual with a true positive for HPV is determined via PCR test, regardless of sampling methods. Sensitivity and specificity were calculated for each approach independently and kappa test was used to assess the consistency between the two approaches., Results: Overall, 211 MSM were recruited at the local clinic from April to October 2020 in Zhuhai, China. The mean age was 31 years old. Only 3% of the participants sought help from healthcare providers during self-sampling. The prevalence of HPV was 49% (103 of 211). Clinician sampling detected 91 of 103 MSM infected with HPV, with a sensitivity of 88.3% (95% CI 80.2 to 93.6) and a specificity of 100.0% (95% CI 95.7 to 100.0). Self-sampling detected 81 of 103 MSM infected with HPV, with a sensitivity of 78.6% (95% CI 69.2 to 85.9) and a specificity of 100.0% (95% CI 95.7 to 100.0). The level of agreement was moderate between clinician sampling and self-sampling (k=0.67)., Conclusions: Self-sampled HPV testing demonstrated comparable accuracy and consistency to clinician sampling among MSM in China. It holds the potential to complement sexual health services especially among key populations., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.)

Advances in plant phenotyping technologies are dramatically reducing the marginal costs of collecting multiple phenotypic measurements across several time points. Yet, most current approaches and best statistical practices implemented to link genetic and phenotypic variation in plants have been developed in an era of single-time-point data. Here, we used time-series phenotypic data collected with an unmanned aircraft system for a large panel of soybean (Glycine max (L.) Merr.) varieties to identify previously uncharacterized loci. Specifically, we focused on the dissection of canopy coverage (CC) variation from this rich data set. We also inferred the speed of canopy closure, an additional dimension of CC, from the time-series data, as it may represent an important trait for weed control. Genome-wide association studies (GWASs) identified 35 loci exhibiting dynamic associations with CC across developmental stages. The time-series data enabled the identification of 10 known flowering time and plant height quantitative trait loci (QTLs) detected in previous studies of adult plants and the identification of novel QTLs influencing CC. These novel QTLs were disproportionately likely to act earlier in development, which may explain why they were missed in previous single-time-point studies. Moreover, this time-series data set contributed to the high accuracy of the GWASs, which we evaluated by permutation tests, as evidenced by the repeated identification of loci across multiple time points. Two novel loci showed evidence of adaptive selection during domestication, with different genotypes/haplotypes favored in different geographic regions. In summary, the time-series data, with soybean CC as an example, improved the accuracy and statistical power to dissect the genetic basis of traits and offered a promising opportunity for crop breeding with quantitative growth curves., (© 2022 The Authors. Journal of Integrative Plant Biology published by John Wiley & Sons Australia, Ltd on behalf of Institute of Botany, Chinese Academy of Sciences.)

The estimation of yield parameters based on early data is helpful for agricultural policymakers and food security. Developments in unmanned aerial vehicle (UAV) platforms and sensor technology help to estimate yields efficiency. Previous studies have been based on less cultivars (<10) and ideal experimental environments, it is not available in practical production. Therefore, the objective of this study was to estimate the yield parameters of soybean (Glycine max (L.) Merr.) under lodging conditions using RGB information. In this study, 17 time point data throughout the soybean growing season in Nanchang, Jiangxi Province, China, were collected, and the vegetation index, texture information, canopy cover, and crop height were obtained by UAV-image processing. After that, partial least squares regression (PLSR), logistic regression (Logistic), random forest regression (RFR), support vector machine regression (SVM), and deep learning neural network (DNN) were used to estimate the yield parameters. The results can be summarized as follows: (1) The most suitable time point to estimate the yield was flowering stage (48 days), which was when most of the soybean cultivars flowered. (2) The multiple data fusion improved the accuracy of estimating the yield parameters, and the texture information has a high potential to contribute to the estimation of yields, and (3) The DNN model showed the best accuracy of training (R 2 =0.66 rRMSE=32.62%) and validation (R 2 =0.50, rRMSE=43.71%) datasets. In conclusion, these results provide insights into both best estimate period selection and early yield estimation under lodging condition when using remote sensing., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Bai, Li, Zhao, Wang, Shao, Guo, Liu, Wang, Li, Guo, Wang, Li, Qiu and Jin.)

Direct oxidation of benzyl alcohol (BzOH) to benzaldehyde (BzH) using O 2 as the oxidant has crucial significance under mild conditions in industrial manufacturing. Herein, an efficient and ecofriendly Pd/O-g-C 3 N 4 heterojunction catalyst using oxygen-doped g-C 3 N 4 (O-g-C 3 N 4 ) with a porous surface as the support is developed for visible light-driven thermocatalytic oxidation of BzOH to BzH by using O 2 as the sole oxidant. The obtained Pd/O-g-C 3 N 4 exhibits visible light-enhanced catalytic performance under visible light irradiation with BzOH conversion and BzH yield of 89.4 and 76.6%, respectively, which are 1.96 and 1.83 times higher than those for the catalyst in the dark. Oxygen doping, as well as Pd deposition, can significantly narrow the band gap of g-C 3 N 4 , thereby enhancing its visible light harvesting ability. The resulting heterojunction interface between Pd nanoparticles and O-g-C 3 N 4 might promote separation of the photogenerated e - -h + pairs. In addition to achieving considerable BzOH conversion and BzH yield, Pd/O-g-C 3 N 4 also exhibits impressive stability. The current study provides potential applications for the green and efficient catalytic oxidation of other alcohols to corresponding aldehydes using O 2 as the oxidant.

Aim: We aimed at developing a fast and accurate method to detect Vibrio mimicus using real-time recombinase polymerase amplification assay., Methods and Results: Specific primers and probe were designed to target V. mimicus haemolysin (vmh) gene. Target DNA was successfully amplified at 41°C within 20 min. The method exhibited a high level of specificity and the sensitivity was 2.1 × 10 2 copies/25 μl or 8.4 copies/μl, which is in line with real-time polymerase chain reaction (PCR). The calibration curve plotted by the second-order polynomial regression showed better than the linear curve, as the correlation coefficient was raised to 0.9907, which suggested that the second-order polynomial regressions might be considered to apply to the quantification of real-time recombinase polymerase amplification (RPA). The limit of detection (LOD) was predicted to be 77 copies/25 μl or 3 copies/μl by a probit model. The limit of quantification (LOQ) was calculated to be 28 copies /25 μl or 1 copies/μl by a receiver operating characteristic (ROC) curve, which firstly make LOQ could be available to real-time RPA. For the performance of the real-time RPA in plasma samples, the detection sensitivity of real-time RPA was as good as the real-time PCR. For pretreatment of plasma samples, the boiling method was better than using kits, as it further shortened the time of the real-time RPA in detecting V. mimicus., Conclusions: The real-time RPA assay developed in our study shows multiple advantages over currently available DNA diagnostic method, including a quicker time-to-result for a single sample, requiring minimal infrastructure and technical support and being tolerant to inhibitors in plasma samples., Significance and Impact of the Study: The real-time RPA assay developed here is a potentially valuable tool for point-of-care (POC) diagnosis of V. mimicus infection in endemic field, especially in the resources-limited settings, as combined with portable devices., (© 2022 Society for Applied Microbiology.)

Development of efficient and robust electrocatalysts for complete oxidation of ethanol is critical for the commercialization of direct ethanol fuel cells. However, the complete oxidation of ethanol suffers from poor efficiency due to the low C1 pathway selectivity. Herein, single-atomic Ir (Ir 1 ) on hcp-PtPb/fcc-Pt core-shell hexagonal nanoplates (PtPb@PtIr 1 HNPs) enclosed by Pt(110) surface with a 7.2% tensile strain is constructed to drive complete electro-oxidation of ethanol. Benefiting from the construction of Ir 1 sites, the PtPb@PtIr 1 HNPs exhibit a Faraday efficiency of 57.93% for the C1 pathway, which is ≈8.3 times higher than that of the commercial Pt/C-JM. Furthermore, the PtPb@PtIr 1 HNPs show a top-ranked electro-activity achieving 45.1-fold and 56.3-fold higher than the specific and mass activities of Pt/C-JM, respectively. Meanwhile, the durability can be significantly enhanced by the construction of Ir 1 sites. Density functional theory calculations indicate that the strong synergy on the PtPb@PtIr 1 HNPs surface significantly promotes the breaking of CC bond of CH 2 CO* and facilitates CO oxidation and suppresses the deactivation of the catalyst. This work offers a unique single-atom approach using low-coordination active sites on shape-controlled nanocrystals to tune the selectivity and activity toward complicated catalytic reactions., (© 2022 Wiley-VCH GmbH.)

In this work, titanium dioxide (TiO2)-incorporated rice straw fiber (RS)/poly(butylene succinate) (PBS) biocomposites were prepared by injection molding with different TiO2 powder loadings. The RS/PBS with 1 wt% TiO2 demonstrated the best mechanical properties, where the flexural strength and modulus increased by 30.34% and 28.39%, respectively, compared with RS/PBS. The non-isothermal crystallization of neat PBS, RS/PBS composites, and titanium-dioxide-incorporated RS/PBS composites was investigated by differential scanning calorimetry (DSC) and X-ray diffraction (XRD). The non-isothermal crystallization data were analyzed using several theoretical models. The Avrami and Mo kinetic models described the non-isothermal crystallization behavior of neat PBS and the composites; however, the Ozawa model was inapplicable. The crystallization temperature (Tc), half-time of crystallization (t1/2), and kinetic parameters (FT) showed that the crystallizability followed the order: TiO2-incorporated RS/PBS composites > RS/PBS > PBS. The RS/PBS with 1 wt% TiO2 showed the best crystallization properties. The Friedman model was used to evaluate the effective activation energy of the non-isothermal crystallization of PBS and its composites. Rice straw fiber and TiO2 acted as nucleating agents for PBS. The XRD results showed that the addition of rice straw fiber and TiO2 did not substantially affect the crystal parameters of the PBS matrix. Overall, this study shows that RS and TiO2 can significantly improve the crystallization and mechanical properties of PBS composites.

Corona Virus Disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is seriously threatening human health. Following SARS-CoV-2 infection, immune cell infiltration creates an inflammatory and oxidative microenvironment, which can cause pneumonia, severe acute respiratory syndrome, kidney failure, and even death. Clinically, a safe and effective treatment strategy remains to be established. Herein, a nano-bait strategy for inhibition of SARS-CoV-2 infection by redirecting viral attack while simultaneously relieving inflammation is developed. Specifically, the nano-bait was based on the exosome-sheathed polydopamine (PDA@Exosome) nanoparticles, which were generated by exocytosis of the PDA nanoparticles from H293T cells. In this approach, PDA@Exosome inherits from the source cells of H293T cells a surface display of ACE2 through pre-engineered expression. The resulting PDA@Exosome can compete with ACE2-expressing epithelial cells for S protein binding, in either the pre-exposure or post-exposure route. Moreover, relying on the ability of PDA to intercept and deactivate radical species, the PDA@Exosome can significantly attenuate the level of inflammatory cytokines by mediating oxidative stress, a major cause of organ injury. Due to its high trapping, multiple antioxidant ability, and good biocompatibility, the HACE2-exosome based nano-bait is a promising robust antiviral nanotherapeutics for the ongoing COVID-19 pandemic.

Novel three-dimensional hierarchical α-calcium sulfate hemihydrate with a straw-sheaf morphology (3D α-HH straw-sheaves) are synthesized successfully in glycerin aqueous solution by a simple one-pot method, using as an efficient adsorbent for Pb 2+ removal from water. The 3D straw-sheaf morphology, that closely depends on the glycerin/water volume ratio (V Gly /V H2O ), can be accurately fabricated only when V Gly /V H2O is not lower than 3/1. 3D α-HH straw-sheaves are generated via multistep-splitting growth coupled with self-assembly. The obtained 3D α-HH straw-sheaves are further used as an adsorbent to remove Pb 2+ from water, exhibiting excellent Pb 2+ removal performance with an equilibrium adsorption capacity of 79.19 mg Pb g α-HH -1 and removal efficiency of 98.98%, that both higher than those of plate- and columnar-like α-HH. Moreover, the experimental adsorption data for the 3D α-HH straw-sheaves is well fitted with pseudo-second-order kinetic model, and the adsorption isotherm is in good agreement with Langmuir model. The Pb 2+ adsorption mechanism is thought to be a chemical adsorption process enforced by chemical bonding and ion exchange. This work demonstrates that 3D α-HH straw-sheaves are highly promising in removing Pb 2+ from wastewater, thereby broadening the research field for the practical application of gypsum-based materials., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Recent studies reported that miR-128 was differentially expressed in cardiomyocytes in response to pathologic stress. However, its function and mechanism remain to be fully elucidated. The aim of the present study was to investigate the role of miR-128 in chronic angiotensin II (Ang II) infusion-induced cardiac remodeling and its underlying mechanism. The cardiac remodeling and heart failure in vivo were established in C57BL/6 mice by chronic subcutaneous Ang II delivery. Knocking down miR-128 was conducted in the hearts of the mice by intravenous injection of HBAAV2/9-miR-128-GFP sponge (miR-128 inhibitor). In vitro experiments of cardiac hypertrophy, apoptosis, and aberrant autophagy were performed in cultured cells after Ang II treatment or transfection of miR-128 antagomir. Our results showed that chronic Ang II delivery for 28 days induced cardiac dysfunction, hypertrophy, fibrosis, apoptosis, and oxidative stress in the mice, while the miR-128 expression was notably enhanced in the left ventricle. Silencing miR-128 in the hearts of mice ameliorated Ang II-induced cardiac dysfunction, hypertrophy, fibrosis apoptosis, and oxidative stress injury. Moreover, Ang II induced excessive autophagy in the mouse hearts, which was suppressed by miR-128 knockdown. In cultured cells, Ang II treatment induced a marked elevation in the miR-128 expression. Downregulation of miR-128 in the cells by transfection with miR-128 antagomir attenuated Ang II-induced apoptosis and oxidative injury probably via directly targeting on the SIRT1/p53 pathway. Intriguingly, we found that miR-128 inhibition activated PIK3R1/Akt/mTOR pathway and thereby significantly damped Ang II-stimulated pathological autophagy in cardiomyocytes, which consequently mitigated cell oxidative stress and apoptosis. In conclusion, downregulation of miR-128 ameliorates Ang II-provoked cardiac oxidative stress, hypertrophy, fibrosis, apoptosis, and dysfunction in mice, likely through targeting on PIK3R1/Akt/mTORC1 and/or SIRT1/p53 pathways. These results indicate that miR-128 inhibition might be a potent therapeutic strategy for maladaptive cardiac remodeling and heart failure., Competing Interests: The authors declare that they have no conflict of interest., (Copyright © 2021 Heqin Zhan et al.)

Achieving high catalytic performance with high CO 2 selectivity is critical for commercialization of direct ethanol fuel cells. Here, we report carbon-supported highly porous Pt 2 Ir alloy nanocrystals (p-Pt 2 Ir/C) for an ethanol oxidation reaction (EOR) that displays nearly 7.2-fold enhancement in mass activity and promotes antipoisoning ability and durability for the EOR as compared with the commercial Pt/C-JM. Moreover, the catalyst exhibits high CO 2 selectivity, 3.4-fold at 0.65 V (vs. SCE) and 4.1-fold at 0.75 V (vs. SCE) higher as compared with the carbon-supported porous Pt nanocrystals (p-Pt/C). The highly porous structure is composed of interconnected one-dimensional (1D) rough branches with an average diameter of only 1.9 nm, largely promoting Pt utilization efficiency and accelerating mass transfer. The 1D rough branch surface exposed many atomic steps/corners endowed with abundant high activity sites. Alloying with Ir can significantly improve the antipoisoning ability, durability, and C-C bond cleavage ability, thereby evidently enhancing its EOR performance.

Crystal phase engineering is a powerful strategy for regulating the performance of electrocatalysts toward many electrocatalytic reactions. Herein we demonstrate that Au@Pd 1 Cu concave nanocubes (CNCs) with an ordered body-centered cubic (bcc) PdCu alloy shell enclosed by many high active high-index facets can be adopted as highly active yet stable electrocatalysts for the ethanol oxidation reaction (EOR). These CNCs are more efficient than other nanocrystals with a disordered face-centered cubic (fcc) PdCu alloy surface and display high mass and specific activities of 10.59 A mg pd -1 and 33.24 mA cm -2 , which are 11.7 times and 4.1 times higher than those of commercial Pd black, respectively. Our core-shell CNCs also exhibit robust durability with the weakest decay in activity after 250 potential-scanning cycles, as well as outstanding antipoisoning ability. Alloying with Cu and the ordered bcc phase surface can provide abundant OH ads species to oxidize carbonaceous poison to avoid catalyst poisoning, and the exposed high-index facets on the surface can act as highly catalytic sites.

Vibrio vulnificus (V. vulnificus) is a Gram-negative bacterium living in warm and salty water. This marine bacterium could produce hemolysin (VVH), which often causes serious gastroenteritis or septicemia when people contact to seawater or seafood containing V. vulnificus. Timely diagnosis is regard as essential to disease surveillance. In this paper, we aimed at developing a quick and sensitive method for the detection of Vibrio vulnificus using real time recombinase polymerase amplification (real time RPA). Specific primers and an exo probe were designed on the basis of the vvhA gene sequence available in GenBank. Target DNA could be amplified and labeled with specific fluorophore within 20 min at 38 °C. The method exhibited a high specificity, only detecting Vibrio vulnificus and not showing cross-reaction with other bacteria. The sensitivity of this method was 2 pg per reaction (20 μL) for DNA, or 200 copies per reaction (20 μL) for standard plasmid. The detection limit (LOD) stated as the target level that would be detected 95% of the time and estimated was 1.58 × 10 2 copies by fit of the probit to the results of 8 replicates in different concentration. For quantitative analysis of the real time RPA, the second order polynomial regression was adopted in our study. The results showed the correlation coefficients were raised above 0.98, which suggested this model might be a better choice for the quantitative analysis of real time RPA compared to the routine linear regression model. For artificially contaminated plasma samples, Vibrio vulnificus could be detected within 16 min by real time RPA at concentration as low as 1.2 × 10 2  CFU/mL or 2.4 CFU per reaction (20 μL). Thus, the real time RPA method established in this study shows great potential for detecting Vibrio vulnificus in the research laboratory and disease diagnosis., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The network security transmission of digital images needs to solve the dual security problems of content and appearance. In this paper, a visually secure image compression and encryption scheme is proposed by combining compressed sensing (CS) and regional energy. The plain image is compressed and encrypted into a secret image by CS and zigzag confusion. Then, according to the regional energy, the secret image is embedded into a carrier image to obtain the final visual secure cipher image. A method of hour hand printing (HHP) scrambling is proposed to increase the pixel irrelevance. Regional energy embedding reduce the damage to the visual quality of carrier image, and the different embedding positions between images greatly enhances the security of the encryption algorithm. Furthermore, the hyperchaotic multi-character system (MCS) is utilized to construct measurement matrix and control pixels. Simulation results and security analyses demonstrate the effectiveness, security and robustness of the propose algorithm.

In the actual image processing process, we often encounter mixed images that contain multiple valid messages. Such images not only need to be transmitted safely, but also need to be able to achieve effective separation at the receiving end. This paper designs a secure and efficient encryption and separation algorithm based on this kind of mixed image. Since chaotic system has the characteristics of initial sensitivity and pseudo-randomness, a chaos matrix is introduced into the compressed sensing framework. By using sequence signal to adjust the chaotic system, the key space can be greatly expanded. In the algorithm, we take the way of parallel transmission to block the data. This method can realize the efficient calculation of complex tasks in the image encryption system and improve the data processing speed. In the decryption part, the algorithm in this paper can not only realize the restoration of images, but also complete the effective separation of images through the improved restoration algorithm., Competing Interests: The authors declare no conflict of interest.

In this paper, a new image encryption transmission algorithm based on the parallel mode is proposed. This algorithm aims to improve information transmission efficiency and security based on existing hardware conditions. To improve efficiency, this paper adopts the method of parallel compressed sensing to realize image transmission. Compressed sensing can perform data sampling and compression at a rate much lower than the Nyquist sampling rate. To enhance security, this algorithm combines a sequence signal generator with chaotic cryptography. The initial sensitivity of chaos, used in a measurement matrix, makes it possible to improve the security of an encryption algorithm. The cryptographic characteristics of chaotic signals can be fully utilized by the flexible digital logic circuit. Simulation experiments and analyses show that the algorithm achieves the goal of improving transmission efficiency and has the capacity to resist illegal attacks.

Molecular characterizations, including microsatellite instability (MSI) and the CpG island methylator phenotype (CIMP) showed strong associations in colorectal carcinoma (CRC) and provided a deeper understanding of the etiology of disease. However, the global relationship between epigenetic alternations and changes in mRNA expression in CRC remains largely undefined, especially regarding the roles of DNA methyltransferases (DNMTs). Here, we conducted a systematic network comparison to explore the global conservation between co-expressed and co-methylated modules. We successfully identified immune-related modules that were regulated by DNMTs and had strong associations with immune-infiltrating neutrophils and dendritic cells in CRC. Moreover, we found that genes in those modules were prognostic for CRC, with 97.1% (168/173) being significantly influenced by DNMTs. Thus, this study resolved an interaction between DNA methylation and mRNA expression through DNMTs. Additionally, we provided evidence that DNMTs control the global hypomethylation of oncogenes, including ALOX5AP and CSF3R that otherwise have high methylation in normal colons. Such genes were also more sensitive to DNMT changes, such as in CRC. Collectively, our analyzes provided a systems biology approach to investigate the association among different molecular phenotypes in diseases.

The uptake of trace elements can be impeded by precipitation in the presence of carbonates and sulfates. The objective of this study was to investigate whether ethylenediaminetetraacetic acid (EDTA) enhances the performance of anaerobic digestion by forming dissolved complexes with Fe 2+ . Batch experiments were performed in this study and acidogenic and methanogenic phases were artificially simulated. EDTA was added to both of phases to examine its effects on Fe bioavailability, metabolic parameters and microbial community structure. The results showed that EDTA significantly accelerated the digestion process in both phases because its addition changed the Fe sorption law and increased Fe-bioavailability. The microbial community structure changed following by the change of Fe-fractions which was determined by EDTA. This study demonstrated that EDTA as ligand could increase the Fe-bioavailability and then reduced or replaced the addition of Fe., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The addition of trace elements to aid anaerobic digestion has already been widely studied. However, the effects of rare trace elements on anaerobic digestion remain unclear. In this study, the effects of Mo, Se and Mn on anaerobic digestion of rice straw were explored. The results showed the methane yield increased by 59.3%, 47.1% and 48.9% in the first 10 days following addition of Mo (0.01 mg/L), Se (0.1 mg/L) and Mn (1.0 mg/L), respectively. Toxic effects and the accumulation of volatile fatty acids (VFAs) were observed when the Se, Mo and Mn concentrations were greater than 100, 1000 and 1000 mg/L, respectively. The half-maximal inhibitory concentrations (IC 50 ) for Se, Mn and Mo were 79.9 mg/L, 773.9 mg/L and 792.3 mg/L, respectively. The addition of trace elements has changed the bacterial structure of the bacteria, which in turn has affected the digestion performance., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Recently, Celastrus orbiculatus ethyl acetate extracts (COE) have been investigated for their anticancer effects on digestive tract tumors. However, the therapeutic effects of COE on esophageal squamous carcinoma cells (ESCC) have not been investigated. In the present study, the effects of COE on the cell cycle and apoptosis of ESCCs were assessed in vitro , and it was revealed that COE treatment triggered G 0 /G 1 cell cycle arrest, and induced DNA damage and apoptosis in a dose-dependent manner in ESCC. Activation of the phosphatidylinositol 3-kinase/protein kinase-B/mechanistic target of rapamycin (mTOR) pathway was also suppressed by COE. Additionally, the combined treatment with COE and rapamycin (an mTOR inhibitor) acted synergistically in ECA-109 cells compared with the treatment with COE or rapamycin alone. These findings extend the understanding of the action of COE and suggest that COE has potential as a treatment option for ESCC as a single treatment or in combination.

A one-step in situ nanoconfined pyrolysis strategy was developed to anchor highly active single Co atoms on the P,N-doped porous carbon@carbon nanotube coaxial nanocables (Co-P,N-CNT), which exhibit remarkable enhanced ORR electrocatalytic activity in both acidic and alkaline media.

Ethnopharmacological Relevance: The traditional Chinese medicine (TCM) herb Celastrus orbiculatus is an important folk medicinal plant in China that has been used as an anti-inflammatory, antitumor, and analgesic in various diseases. The ethyl acetate extract of C. orbiculatus (C. orbiculatus extract, COE) was reported to show significant antitumor effects. However, no study in China or abroad has reported the effect and mechanism of COE in triggering apoptosis of gastric cancer (GC) cells., Aim of Study: To further uncover the molecular mechanism underlying COE's apoptotic and anti-proliferative effects and lay a foundation for the development of novel, effective antitumor TCM agents., Materials and Methods: The effect of COE on AGS and BGC-823 GC cell viability was examined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis of AGS and BGC-823 cells induced by COE was analyzed using flow cytometry and a mitochondrial membrane potential assay kit (JC-1). The proliferating GC cells were identified and examined using a 5-bromo-2'-deoxyuridine (BrdU) staining kit and flow cytometric analysis. A western blot assay was used to detect the effect of COE on apoptosis-related proteins, B-cell lymphoma-2 (Bcl-2), Bcl-extra-large (Bcl-xL), Bcl-2-like protein 12 (Bcl-L12), Bcl-2-associated X protein (Bax), and caspase as well as proliferation-related proteins, phosphoinositide 3-kinase (PI3K)/Akt/mechanistic target of rapamycin (mTOR)/p70s6k. Transmission electron microscopy (TEM) and an animal imaging technique were used to evaluate the microstructure of apoptotic GC cells and the effect of COE on tumor cell growth in vivo, respectively., Results: The results indicate that COE significantly inhibited proliferation and induced apoptosis of GC AGS and BGC-823 cell lines both in vivo and in vitro. COE significantly decreased the cell mitochondrial membrane potential. Moreover, COE downregulated the levels of Bcl-2, Bcl-xL, and PI3K/Akt/mTOR/p70s6k while those of Bax and caspase were upregulated. More interestingly, COE altered the microstructure of the mitochondria., Conclusion: All these data collectively indicate that COE not only has significant antiproliferative effects but also has both in vivo and in vitro apoptotic effects. In addition, COE altered the structure and function of the mitochondria, which is another potential pathway for the antitumor activity of COE. These findings may provide a basis for the development of new anticancer TCM candidates., (Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.)

Aim: To investigate the effect of mechanical stress on periostin and semaphorin-3A expression in a murine model of postmenopausal osteoporosis and in osteoblast-like MC3T3-E1 cells., Materials and Methods: Female mice were divided into three groups and treated with a sham operation, ovariectomy (OVX) or OVX plus treadmill training (OVX+Run). After 10 weeks, tibias were used for histological analysis. MC3T3-E1 cells were burdened by mechanical stress using a centrifuge or were treated with periostin, and the production of biologically-active semaphorin-3A was examined in vitro., Results: In OVX+Run group tibias, the number of tartrate-resistant acid phosphatase-positive osteoclasts was lower than in the OVX group, and the expression of periostin and semaphorin-3A was higher. In MC3T3-E1 cells, centrifugal stress significantly increased periostin and semaphorin-3A mRNA expression. Treatment with periostin increased the semaphorin-3A level., Conclusion: We speculate that mechanical load may increase periostin production in osteoblasts, and periostin may inhibit osteoclast differentiation by its effects on semaphorin-3A. Our results support the concept of a positive correlation between exercise and inhibition of osteoclasts in post-menopausal osteoporosis., (Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Metal-organic frameworks (MOFs) are of great interest as potential electrochemically active materials. However, few studies have been conducted into understanding whether control of the shape and components of MOFs can optimize their electrochemical performances due to the rational realization of their shapes. Component control of MOFs remains a significant challenge. Herein, we demonstrate a solvothermal method to realize nanostructure engineering of 2D nanoflake MOFs. The hollow structures with Ni/Co- and Ni-MOF (denoted as Ni/Co-MOF nanoflakes and Ni-MOF nanoflakes) were assembled for their electrochemical performance optimizations in supercapacitors and in the oxygen reduction reaction (ORR). As a result, the Ni/Co-MOF nanoflakes exhibited remarkably enhanced performance with a specific capacitance of 530.4 F g -1 at 0.5 A g -1 in 1 M LiOH aqueous solution, much higher than that of Ni-MOF (306.8 F g -1 ) and ZIF-67 (168.3 F g -1 ), a good rate capability, and a robust cycling performance with no capacity fading after 2000 cycles. Ni/Co-MOF nanoflakes also showed improved electrocatalytic performance for the ORR compared to Ni-MOF and ZIF-67. The present work highlights the significant role of tuning 2D nanoflake ensembles of Ni/Co-MOF in accelerating electron and charge transportation for optimizing energy storage and conversion devices.

Constitutive photomorphogenesis 9 signalosome (CSN) consists of a total of eight subunits (CSN1-CSN8) in mammalian cells. CSN6 may promote carcinogenesis by positively regulating v‑myc avian myelocytomatosis viral oncogene homolog (Myc) and MDM2 proto‑oncogene stability, and is regarded as a potential target for cancer therapy. Quercetin has a substantial anticancer effect on various human cancer cells. The present study investigated the effects of quercetin on HT-29 human colorectal cancer cell viability, apoptosis and cell cycle arrest using an MTT assay, flow cytometry, transmission electron microscopy and western blotting. It was determined that quercetin inhibited HT‑29 cell viability in a dose‑dependent manner. Cell shrinkage, chromatin condensation and nuclear collapse were observed in the 50, 100 and 200 µM quercetin groups. The exposure of HT‑29 cells to quercetin led to significant cell cycle arrest in the S‑phase. Western blot analysis revealed that quercetin reduced the protein expression levels of phosphorylated-Akt and increased CSN6 protein degradation; therefore, affecting the expression levels of Myc, p53, B‑cell lymphoma 2 (Bcl‑2) and Bcl‑2 associated X protein. The overexpression of CSN6 reduced the effect of quercetin treatment on HT‑29 cells, suggesting that quercetin‑induced apoptosis may involve the Akt‑CSN6‑Myc signaling axis in HT‑29 cells.

Celastrus orbiculatus is used as a folk medicine in China for the treatment of numerous diseases. The ethyl acetate extract of Celastrus orbiculatus (COE) also displays a wide range of anti-cancer activities in the laboratory. However, the effectiveness of COE-induced autophagy and its mechanism of action in colorectal cancer cells have not been investigated thus far. The present study analyzed the effect of COE on HT-29 cell viability, apoptosis and autophagy using MTT assay, flow cytometry, transmission electron microscopy and western blotting. Additionally, the autophagy inhibitor 3-methyladenine and the autophagy inducer rapamycin were used to further explore the effects of COE-induced autophagy in HT-29 cells. The present study also examined whether the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/Akt/mechanistic target of rapamycin (mTOR)/p70 ribosomal protein S6 kinase (p70S6K) signaling pathway was involved in the regulation of COE-induced autophagy. The results revealed that COE inhibited HT-29 cell proliferation and decreased cell survival in a time- and dose-dependent manner, and that COE possessed the ability to induce both apoptosis and autophagy in HT-29 cells. Furthermore, autophagy and apoptosis induced by COE synergized to inhibit colorectal cancer growth. In addition, COE treatment decreased the phosphorylation of Akt and its downstream effectors mTOR and p70S6K. Taken together, these results demonstrate that both autophagy and apoptosis were activated during COE treatment of HT-29 cells, and that COE-induced autophagy decreases the viability of HT-29 cells via a mechanism that may depend on the PI3K/Akt/mTOR/p70S6K signaling pathway. Furthermore, compounds that induce autophagy administered in combination with COE may be an attractive strategy for enhancing the anti-tumor potency of COE in colorectal cancer.

An efficient copper-TEMPO-catalyzed one-pot synthesis of α-ketoamides from phenethyl alcohol derivatives was developed firstly. Moreover, molecular oxygen in open air was employed as the oxidant with a broad substrate scope, which makes this methodology more practical. Based on some control experiments, a plausible mechanism was proposed.

Crude okra (Abelmoschus esculentus L.) polysaccharide (RPS) was obtained by water extraction and alcohol precipitation. Three purified fractions of RPS, designated RPS-1, RPS-2, and RPS-3, were fractioned by diethylaminoethyl (DEAE)-cellulose chromatography. Their molecular weights, monosaccharide compositions, infrared (Fourier transform infrared [FT-IR]) spectra, and nuclear magnetic resonance (NMR) spectra were analyzed. Their immunomodulatory activity was evaluated with an in vitro cell model (RAW264.7 cells). In vivo immunomodulatory activity of RPS-2 was evaluated in normal and cyclophosphamide-induced immunosuppressed mice. The results showed that the molecular weights of RPS-1, RPS-2, and RPS-3 were 600, 990, and 1300 kDa, respectively. RPS-1 and RPS-2 were mainly composed of galactose, rhamnose, galacturonic acid, and glucuronic acid, while RPS-3 was mainly composed of galactose, rhamnose, galacturonic acid, glucuronic acid, and glucose. FT-IR and NMR spectrum data indicated a rhamnogalacturonan I characteristic of polysaccharide. Both RPS and its purified fractions RPS-1, RPS-2, and RPS-3 significantly increased RAW264.7 cell proliferation, nitric oxide (NO) production, inducible nitric oxide synthase (iNOS) expression, and tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and interleukin (IL)-10 secretion (P < .05). The purified fraction RPS-2 also increased the spleen index, splenocyte proliferation, and cytokine secretion in vivo. These results indicate that okra polysaccharides may potentially serve as novel immunomodulators.

An economic and practical AIBN-initiated aerobic oxidation system of benzylic alcohols, hetero aryl alcohols and allyl alcohols was developed for the first time. Moderate to excellent yields were obtained with a broad substrate scope. Moreover, a proposed mechanism of a radical process was assumed and confirmed by the key intermediate detected.

Hepatitis E virus- (HEV-) mediated hepatitis has become a global public health problem. An important regulatory protein of HEV, ORF3, influences multiple signal pathways in host cells. In this study, to investigate the function of ORF3 from the swine form of HEV (SHEV), high-throughput RNA-Seq-based screening was performed to identify the differentially expressed genes in ORF3-expressing HepG2 cells. The results were validated with quantitative real-time PCR and gene ontology was employed to assign differentially expressed genes to functional categories. The results indicated that, in the established ORF3-expressing HepG2 cells, the mRNA levels of CLDN6, YLPM1, APOC3, NLRP1, SCARA3, FGA, FGG, FGB, and FREM1 were upregulated, whereas the mRNA levels of SLC2A3, DKK1, BPIFB2, and PTGR1 were downregulated. The deregulated expression of CLDN6 and FREM1 might contribute to changes in integral membrane protein and basement membrane protein expression, expression changes for NLRP1 might affect the apoptosis of HepG2 cells, and the altered expression of APOC3, SCARA3, and DKK1 may affect lipid metabolism in HepG2 cells. In conclusion, ORF3 plays a functional role in virus-cell interactions by affecting the expression of integral membrane protein and basement membrane proteins and by altering the process of apoptosis and lipid metabolism in host cells. These findings provide important insight into the pathogenic mechanism of HEV.

Aim of the Study: Celastrus orbiculatus has been used as a folk medicine in China for the treatment of many diseases. In the laboratory, the ethyl acetate extract of Celastrus orbiculatus (COE) displays a wide range of anticancer functions. However, the inhibition of the metastasis mechanism of COE in gastric cancer cells has not been investigated so far. The present study was undertaken to determine if the antimetastatic effects of COE were involved in inhibition of the epithelial-mesenchymal transition (EMT) of human gastric adenocarcinoma SGC-7901 cells., Methods: The adhesion, invasion, and migration of SGC-7901 cells were determined by COE treatment in vitro, using Matrigel-coated plate, transwell membrane chamber, and wound healing models, respectively. In vivo, the growth-inhibiting and antimetastatic effects of COE on the nude mice model of gastric cancer were tested and the mechanisms were explored. The expression of EMT markers and nuclear factor κB (NF-κB)/Snail signaling pathway were evaluated by using western blotting and immunohistochemistry., Results: Treatment with COE dose-dependently inhibited the proliferation, adhesion, invasion, and migration of SGC-7901 cells in vitro, which was realized by enhancing the expression of E-cadherin and reducing N-cadherin and vimentin expression. Moreover, COE suppressed the activation of NF-κB/Snail signaling pathway induced by tumor necrosis factor-α. In addition, COE effectively suppressed tumor growth and metastasis in the nude mice model due to reduced expression of N-cadherin, vimentin, NF-κB p65, and Snail and increased expression of E-cadherin in the tumor tissues., Conclusion: Our findings provided new evidence that COE is an effective inhibitor of metastatic potential of SGC-7901 cells through suppression of EMT and NF-κB/Snail signal pathway. Based on these findings, COE may be considered a novel anticancer agent for the treatment of metastasis in gastric cancer., (© The Author(s) 2015.)

Lipopolysaccharide (LPS) stimulates macrophages to release proinflammatory cytokines. MicroRNAs (miRNAs) are short noncoding RNAs that are involved in inflammatory reaction. Our previously study identified the downregulated expression of mmu-miR-27a-5p in RAW267.4 cells treated with LPS. To dissect the mechanism that mmu-miR-27a-5p regulates target genes and affects proinflammatory cytokine secretion more clearly, based on previous bioinformatics prediction data, one of the potential target genes, MCPIP1 was observed to be upregulated with qRT-PCR and western blot. Luciferase reporter assays were performed to further confirm in silico prediction and determine that MCPIP1 is the target of mmu-miR-27-5p. The results suggested that mmu-miR-27a-5p directly targeted the 3'-UTR of MCPIP1 and the interaction between mmu-miR-27-5p and the 3'-UTR of MCPIP1 is sequence-specific. MCPIP1 overexpression decreased the secretion of IL-6, IL-1β, and IL-10 in macrophage cells stimulated with LPS. Our findings may provide the important information for the precise roles of mmu-miR-27a-5p in the macrophage inflammatory response to LPS stimulation in the future.

Background: Celastrus orbiculatus has been used as a folk medicine in China for the treatment of many diseases. In the laboratory, the ethyl acetate extract of Celastrus orbiculatus (COE) displays a wide range of anticancer functions. However, the inhibition of the metastasis mechanism of COE in gastric cancer cells has not been investigated so far., Methods: The present study was undertaken to determine if the anti-metastasis effect of COE was involved in inhibiting of epithelial-mesenchymal transition (EMT) of human gastric adenocarcinoma SGC-7901 cells. In vitro, a well-established experimental EMT model involving transforming growth factor β1 (TGF-β1) was applied. Viability, invasion and migration, protein and mRNA expression of tumor cells were analyzed by MTT assay, transwell assay, western blot and real-time PCR, respectively. The molecular targets of COE in SGC-7901 cells were investigated by two-dimensional gel electrophoresis (2-DE) and MALDI-TOF-TOF mass spectrometer. Overexpression of heat shock protein 27 (HSP27) was performed by transfected with the recombinant retroviral expression plasmid. In vivo, the anti-metastasis mechanisms of COE in the peritoneal gastric cancer xenograft model was explored and the effect was tested., Results: The non-cytostatic concentrations of COE effectively inhibited TGF-β1 induced EMT process in SGC-7901 cells, which is characterized by prevented morphological changes, increased E-cadherin expression and decreased Vimentin, N-cadherin expression. Moreover, COE inhibited invasion and migration induced by TGF-β1. Using a comparative proteomics approach, four proteins were identified as differently expressed, with HSP27 protein being one of the most significantly down-regulated proteins induced by COE. Moreover, the activation of nuclear factor κB (NF-κB)/Snail signaling pathway induced by tumor necrosis factor-α (TNF-α) was also attenuated under the pretreatment of COE. Interestingly, overexpression of HSP27 significantly decreases the inhibitory effect of COE on EMT and the NF-κB/Snail pathway. Furthermore, COE significantly reduced the number of peritoneal metastatic nodules in the peritoneal gastric cancer xenograft model., Conclusions: Taken together, these results suggest that COE inhibits the EMT by suppressing the expression of HSP27, correlating with inhibition of NF-κB/Snail signal pathways in SGC-7901 cells. Based on these results, COE may be considered a novel anti-cancer agent for the treatment of metastasis in gastric cancer.

Background: Invasion of fibroblast-like synoviocytes (FLSs) is critical in the pathogenesis of rheumatoid arthritis (RA). The metalloproteinases (MMPs) and activator of nuclear factor-kappa B (NF-κB) pathway play a critical role in RA-FLS invasion induced by interleukin-1 beta (IL-1β) and tumour necrosis factor-alpha (TNF-α). The present study aimed to explore the anti-invasive activity and mechanism of Celastrus orbiculatus extract (COE) on IL-1β and TNF-α combination-stimulated human RA-FLSs., Methods: We investigated the effect of COE on IL-1β and TNF-α combination-induced FLS invasion as well as MMP expression and explored upstream signal transduction., Results: COE suppressed IL-1β and TNF-α combination-stimulated FLSs invasion by inhibiting MMP-9 expression and activity. Furthermore, our results revealed that COE inhibited the transcriptional activity of MMP-9 by suppression of the binding activity of NF-κB in the MMP-9 promoter, and inhibited IκBα phosphorylation and nuclear translocation of NF-κB., Conclusions: COE inhibits IL-1β and TNF-α combination-induced FLSs invasion by suppressing NF-κB-mediated MMP-9 expression.

A cluster of differentiation antigen 14 (CD14) is involved in lipopolysaccharide (LPS)-induced proinflammatory cytokine release and LPS-induced septic shock. MicroRNAs (miRNAs) are short non-coding RNAs that are involved in the epigenetic regulation of cellular process and bacterial infection. Our previous study indicated that siRNA against CD14 effectively inhibited LPS-induced tumor necrosis factor alpha, chemokine (C-X-C motif) ligand 2, interleukin-6 release, and NO production. To identify miRNAs which are affected by CD14 gene silencing and dissect the mechanisms of the attenuating of LPS-induced damaging immune activation more clearly, based on the CD14 knockdown RAW264.7 macrophage cell line established in our previous study, miRNAs expression profiling of CD14 knockdown RAW264.7 cells were analyzed with miRNA microarray and validated by qRT-PCR, the potential targets were predicted and subjected to gene ontology (GO) pathway and biological processes analysis. We demonstrated for the first time that CD14 knockdown significantly changed the expression of 199a-3p, miR-199a-5p, and miR-21-5p in RAW264.7 cells, and significantly enriched GO terms in the predicted target genes of these miRNAs were apoptosis process, immune response, inflammatory response, innate immune response, anti-apoptosis, cytokine production, and cytokine-mediated signaling pathway. These findings may improve our understanding about functional mechanism of miRNAs in the attenuating of LPS-induced damaging immune activation more clearly.

Hepatitis E virus (HEV) has emerged as an important cause of epidemic and sporadic acute viral hepatitis worldwide, which is a major public health challenge. A better understanding of the interaction between the virus and the host cell would be very helpful for its therapy. Swine HEV (SHEV) open reading frame 3 (ORF3) is a regulatory protein that alters the activity of selected transcription factors and cytoplasmic signaling pathways. MicroRNAs (miRNAs) are potent post-transcriptional regulators of protein-coding genes and represent an interesting lead to study SHEV infection and to identify new therapeutic targets. To explore how SHEV ORF3 affects miRNAs in host cells, we used miRNA array analysis to compare the expression patterns of miRNAs in stable cell lines that expressed or did not express SHEV ORF3. We found a significant down-regulation of miR-221 and -222 in ORF3 expressing human embryonic kidney 293 cell line. Among the 116 candidate targets genes of miR-221 and -222 that we detected in silico, we demonstrated that the expression of the cyclin-dependent kinase inhibitor 1B, also named p27(kip1), was directly regulated by these miRNAs. We hypothesize that SHEV ORF3-induced miR-221/222 downregulation enhances p27(kip1) expression in HEK293 cells. This provides new avenues for future exploration of the precise roles of miRNAs in SHEV infection.

Invasion of fibroblast-like synoviocytes (FLSs) is critical in the pathogenesis of rheumatoid arthritis (RA). The metalloproteinases (MMPs) and activator of Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) pathway play a critical role in RA-FLS invasion induced by lipopolysaccharide (LPS). The present study aimed to explore the anti-invasive activity of celastrol on LPS-stimulated human RA-FLSs, and to elucidate the mechanism involved. We investigated the effect of celastrol on LPS-induced FLS migration and invasion as well as MMP expression and explored the upstream signal transduction. Results showed that celastrol suppressed LPS-stimulated FLS migration and invasion by inhibiting MMP-9 expression and activity. Furthermore, our results revealed that celastrol inhibited the transcriptional activity of MMP-9 by suppressing the binding activity of NF-κB in the MMP-9 promoter, and suppressed the TLR4/MyD88/NF-κB pathway. Administration of celastrol (0.5 mg/kg and 1 mg/kg, intraperitoneally) daily for 3 weeks in a collagen-induced arthritis rat model markedly alleviated the clinical signs, synovial hyperplasia and inflammatory cell infiltration of joints. In conclusion, celastrol might inhibit FLS migration and invasion induced by LPS by suppressing TLR4/NF-κB-mediated MMP-9 expression, providing a theoretical foundation for the clinical treatment of RA with celastrol.

Both hypoxia and interleukin-17A (IL-17A) promote the migration and invasion of fibroblast-like synoviocytes (FLSs), which are critical for the pathogenesis of rheumatoid arthritis (RA). However, the biochemical pathways regulating IL-17A combined with hypoxia are not well defined. In this study, we found that co-stimulating RA-FLSs with IL-17A and hypoxia did not appear to promote the epithelial-mesenchymal transition (EMT), but did increase cell motility. We further showed that a proinvasive effect of IL-17A on FLSs under hypoxia might be through upregulation of matrix metalloproteinase 2 (MMP2) and MMP9. Moreover, IL-17A-induced expression of MMP2 and MMP9 under hypoxia was accompanied by increased activation of nuclear factor-κB (NF-κB)/hypoxia-inducible factor-1α (HIF-1α). Knockdown or inhibition of HIF-1α and NF-κB by small interfering RNA or specific small molecule inhibitors blocked IL-17A-mediated and hypoxia-mediated MMP2 and MMP9 expression, cell migration, and invasion. In addition, the inhibition of NF-κB led to a marked decrease in the expression of HIF-1α, which indicated that IL-17A activated HIF-1α via the NF-κB pathway in hypoxia. Taken together, our observations suggest a synergetic effect of IL-17A and hypoxia that might contribute to the migration and invasion of RA-FLSs by upregulating the expression of MMP2 and MMP9 by activation of the NF-κB/HIF-1α pathway., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Objective: To investigate the apoptotic effects and underlying molecular mechanisms of Celastrus orbiculatus (C. orbiculatus) extract in human hepatocellular carcinoma cells., Methods: Human hepatocellular carcinoma cells (HCCLM6) were treated with C. orbiculatus extract (COE) at different nontoxic concentrations (10, 20, 40, 80, and 160 microg/mL). The effect of COE on HCCLM6 viability was examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Cellular apoptosis following COE treatment was assessed by flow cytometry and western blot analysis., Results: COE significantly inhibited cell viability and induced apoptosis of HCCLM6 cells in a dose-dependent manner. Apoptosis was accompanied by increased Bax expression and decreased Bcl-2 expression. In addition, COE treatment led to the release of cytochrome c, activation of caspase-3, and cleavage of poly (ADP-ribose) polymerase (PARP). Furthermore, activation of extracellular signal-regulated kinase (ERK), p38 kinase, and c-Jun N-terminal kinase (JNK) phosphorylation, and down-regulation of Akt phosphorylation was observed., Conclusion: COE induces mitochondrial-mediated, caspase-dependent apoptosis in HCCLM6 cells, which might be attributed to the activation of mitogen-activated protein kinase (MAPK) and inhibition of Akt signaling pathways. These data suggest that COE may be a potential treatment for human hepatocellular carcinoma.

To explore the potential role of norepinephrine (NE) and nitric oxide (NO) in activities of rat hypothalamus arginine vasopressin (AVP) neurons in response to immune challenge, we observed the effect of prazosin, an antagonist of alpha1-adrenergic receptor, and the specific nitric oxide synthase (NOS) inhibitor N(w)nitro-L-arginine-methylester (L-NAME) on the Fos expression in AVP neurons induced by systemic lipopolysaccharide (LPS) using double immunohistochemistry. Intravenous (i.v.) injection of LPS induced Fos expression in AVP neurons mainly in the hypothalamus paraventricular nucleus (PVN) and in the supraoptic nucleus (SON). The percentage of Fos-positive AVP neurons was dose-dependent. Pretreatment with prazosin (5 mg/kg) effectively suppressed the Fos expression induced by LPS (5 microg/kg), whereas L-NAME (30 mg/kg) did not influence the Fos expression in the AVP neurons induced by LPS (0.25, 0.5, 1, 5 microg/kg). Our results suggest that the activation of central AVP neurons caused by systemic LPS may be mediated by NE through alpha1-adrenergic receptors, but could not be changed by NO.

Clonidine, an alpha(2)-adrenoceptor agonist, has been reported to inhibit gastric and small intestinal motility in rats. Whether clonidine also inhibits colonic motility is still not clear. The aim of this study was to examine the effect of clonidine on colonic motility and its possible site of action in adult Wistar rats. Colonic motilities in anesthetized rats in vivo or motilities of the isolated colon of rats in vitro were recorded. Clonidine was administered intravenously (i.v.) and intracerebroventricularly (i.c.v.) in vivo while bath administration was used in in vitro study. Clonidine i.v. or i.c.v. significantly inhibited colonic motility. This inhibitory effect was antagonized by pre-administration of yohimbine, an alpha(2)-adrenoceptor antagonist, but not by pre-administration of prazosin, an alpha(1)-adrenoceptor antagonist. Also, we have unpublished data indicating that the sympathectomy antagonized the inhibitory effect of systemically administered clonidine. A significant depression of colonic motility on the isolated colon was induced by bath administration of noradrenaline, while no such inhibition was seen by clonidine. The results of the present study suggested that clonidine inhibits colonic motility in rats through activation of central alpha(2)-adrenergic receptor.

The aim of the present study was to investigate the effect of indomethacin on the Fos expression in arginine vasopressin (AVP)-containing neurons in the hypothalamus and tyrosine hydroxylase (TH)-containing neurons in the locus coeruleus (LC) using dual-labeled immunohistochemistry. In the hypothalamus, intraperitoneal (i.p) injection of different doses [2.5 microg/100 g, 125 microg/100 g body weight (b.w.)] of lipopolysaccharide (LPS) induced a significant Fos expression in AVP neurons in the supraoptic nucleus (SON), the magnocellular division (mPVN) and the parvocellular division (pPVN) of the paraventricular nucleus (PVN). Pretreatment with the cyclooxygenase inhibitor indomethacin (0.8 mg/100 g b.w.) significantly blocked the Fos expression in these AVP neurons induced by a low dose of LPS (2.5 microg/100 g) but had no effect on the Fos expression induced by a high dose of LPS (125 microg/100 g). Similarly, in the brain stem, a large number of TH-positive neurons in the LC expressed Fos after administration of either dose of LPS. Indomethacin prevented the Fos expression induced only by a low dose of LPS, but not by a high dose of LPS. These results suggest that the activation of AVP neurons in PVN and SON and TH neurons in LC response to immune challenge might be mediated-at least partially-by prostaglandins.