Preparation and High-Temperature Resistance Properties of Phenolic Resin/Phosphate Hybrid Coatings.

In this study, a novel fabrication method was used to synthesize phenolic resin/phosphate hybrid coatings using aluminum dihydrogen phosphate (Al(H2PO4)3, hereafter denoted as Al), SC101 silica sol (Si) as the primary film-forming agent, and phenolic resin (PF) as the organic matrix. This approach culminated in the formation of Al+Si+PF organo-inorganic hybrid coatings. Fourier-transform infrared spectroscopy (FT-IR) and X-ray photoelectron spectroscopy (XPS) results confirmed the successful integration of hybrid structures within these coatings. The crystalline structure of the coatings post-cured at various temperatures was elucidated using X-ray diffraction (XRD). Additionally, the surface and cross-sectional morphologies were meticulously analyzed using scanning electron microscopy (SEM), offering insights into the microstructural properties of the coatings. The coatings' porosities under diverse thermal and temporal regimes were quantitatively evaluated using advanced image processing techniques, revealing a significant reduction in porosity to a minimum of 5.88% following a thermal oxidation process at 600 °C for 10 h. The antioxidant efficacy of the phosphate coatings was rigorously assessed through cyclic oxidation tests, which revealed their outstanding performance. Specifically, at 300 °C across 300 h of cyclic oxidation, the weight losses recorded for phosphate varnish and the phenolic resin-infused phosphate coatings were 0.15 mg·cm-2 and 0.09 mg·cm-2, respectively. Furthermore, at 600 °C and over an identical period, the weight reduction was noted as 0.21 mg·cm-2 for phosphate varnish and 0.085 mg·cm-2 for the hybrid coatings, thereby substantiating the superior antioxidation capabilities of the phenolic resin hybrid coatings in comparison to the pure phosphate varnish.

Corrosion Behavior of Ni-Cr Alloys with Different Cr Contents in NaCl-KCl-MgCl2.

This study investigates the corrosion behavior of Ni-Cr binary alloys, including Ni-10Cr, Ni-15Cr, Ni-20Cr, Ni-25Cr, and Ni-30Cr, in a NaCl-KCl-MgCl2 molten salt mixture through gravimetric analysis. Corrosion tests were conducted at 700 °C, with the maximum immersion time reaching up to 100 h. The corrosion rate was determined by measuring the mass loss of the specimens at various time intervals. Verifying corrosion rates by combining mass loss results with the determination of element dissolution in molten salts using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). Detailed examinations of the corrosion products and morphology were conducted using X-ray diffraction (XRD) and scanning electron microscopy (SEM). Micro-area elemental analysis on the corroded surfaces was performed using an energy dispersive spectrometer (EDS), and the elemental distribution across the corrosion cross-sections was mapped. The results indicate that alloys with lower Cr content exhibit superior corrosion resistance in the NaCl-KCl-MgCl2 molten salt under an argon atmosphere compared to those with higher Cr content; no corrosion products were retained on the surfaces of the lower Cr alloys (Ni-10Cr, Ni-15Cr). For the higher Cr alloys (Ni-20Cr, Ni-25Cr, Ni-30Cr), after 20 h of corrosion, a protective layer was observed in certain areas. The formation of a stable Cr2O3 layer in the initial stages of corrosion for high-Cr content alloys, which reacts with MgO in the molten salt to form a stable MgCr2O4 spinel structure, provides additional protection for the alloys. However, over time, even under argon protection, the MgCr2O4 protective layer gradually degrades due to chloride ion infiltration and chemical reactions at high temperatures. Further analysis revealed that chloride ions play a pivotal role in the corrosion process, not only facilitating the destruction of the Cr2O3 layer on the alloy surfaces but also possibly accelerating the corrosion of the metallic matrix through electrochemical reactions. In conclusion, the corrosion behavior of Ni-Cr alloys in the NaCl-KCl-MgCl2 molten salt environment is influenced by a combination of factors, including Cr content, chloride ion activity, and the formation and degradation of protective layers. This study not only provides new insights into the corrosion resistance of Ni-Cr alloys in high-temperature molten salt environments but also offers significant theoretical support for the design and optimization of corrosion-resistant alloy materials.

Insect Antimicrobial Peptides as Guardians of Immunity and Beyond: A Review.

Antimicrobial peptides (AMPs), as immune effectors synthesized by a variety of organisms, not only constitute a robust defense mechanism against a broad spectrum of pathogens in the host but also show promising applications as effective antimicrobial agents. Notably, insects are significant reservoirs of natural AMPs. However, the complex array of variations in types, quantities, antimicrobial activities, and production pathways of AMPs, as well as evolution of AMPs across insect species, presents a significant challenge for immunity system understanding and AMP applications. This review covers insect AMP discoveries, classification, common properties, and mechanisms of action. Additionally, the types, quantities, and activities of immune-related AMPs in each model insect are also summarized. We conducted the first comprehensive investigation into the diversity, distribution, and evolution of 20 types of AMPs in model insects, employing phylogenetic analysis to describe their evolutionary relationships and shed light on conserved and distinctive AMP families. Furthermore, we summarize the regulatory pathways of AMP production through classical signaling pathways and additional pathways associated with Nitric Oxide, insulin-like signaling, and hormones. This review advances our understanding of AMPs as guardians in insect immunity systems and unlocks a gateway to insect AMP resources, facilitating the use of AMPs to address food safety concerns.

Robust construction of konjac glucomannan/polylactic acid nanofibrous films incorporated with carvacrol via microfluidic blow spinning for food packaging.

In recent years, the application of biopolymer-based nanofibers prepared via microfluidic blow spinning (MBS) for food packaging has continuously increased due to their advantages of biocompatibility, biodegradability, and safety. However, the poor spinnability, undesirable water barrier capacity, and loss of antibacterial and antioxidant properties of biopolymer-based nanofibers strictly restrict their real-world applications. In this work, carvacrol (CV) incorporated konjac glucomannan (KGM)/polylactic acid (PLA) nanofibrous films (KP-CV) were produced by MBS. The FTIR spectra and XRD analysis revealed the hydrogen bonding interactions among CV, PLA, and KGM, thus significantly improving the TS of KP-CV nanofibrous films from 0.23 to 1.27 MPa with increased content of CV from 0 % to 5 %. Besides, KP-CV nanofibrous films showed improved thermal stability, excellent hydrophobicity (WCA: 128.19°, WVP: 1.02 g mm/m2 h kPa), and sustained release of CV combined with good antioxidant activities (DPPH radical scavenging activity: 77.51 ± 1.57 %), and antibacterial properties against S. aureus (inhibition zone: 26.33 mm) and E. coli (inhibition zone: 22.67 mm). Therefore, as prepared KP-CV nanofibrous films can be potentially applied as packaging materials for the extended shelf life of cherry tomatoes.

Multifunctional Sensors Made with Conductive Microframework and Biomass Hydrogel for Detecting Packaging Pressure and Food Freshness.

Food packaging detection devices have attracted attention to optimize storage situations and reduce food spoilage. However, low-cost and highly sensitive multifunctional sensors for detecting both food freshness and packaging pressure are still lacking. In this study, a multifunctional sensor was developed consisting of a MXene coated alcohol-soluble polyurethane fiber network (MXene/APU) and composite biohydrogel films made of konjac glucomannan, chitosan, and blueberry anthocyanin (KCB). Based on the pressure sensitivity of MXene/APU and the color changes of KCB in response to pH values, the sensor can detect internal package bulging, external squeezing, and food deterioration. The pressure sensor shows a sensitivity of 1.16 kPa-1, a response time of 200 ms, a wide strain range of 1092%, and stability over multiple loops. The pressure sensor could detect human motion and identify surface morphologies. The excellent sensor performance was attributed to the porous structure and large specific surface area of microfiber networks, conductivity of MXene nanosheets, and protective effect of KCB films coated on the conductive membrane. Besides, the microfluidic blow-spinning method used to prepare microfiber networks showed the advantages of low energy consumption and high production efficiency. Based on the color changes of blueberry anthocyanin loaded in KCB films in response to pH, the sensor realized sensitive spoilage detection of food containing protein. This study provides a new multifunctional food packaging sensing device and a greater understanding of the optimization and application of related devices.

The Effects of Diet on the Immune Responses of the Oriental Armyworm Mythimna separata.

Nutrients can greatly affect host immune defenses against infection. Possessing a simple immune system, insects have been widely used as models to address the relationships between nutrition and immunity. The effects of high versus low protein-to-carbohydrate ratio (P:C) diets on insect immune responses vary in different studies. To reveal the dietary manipulation of immune responses in the polyphagous agricultural pest oriental armyworm, we examined immune gene expression, phenoloxidase (PO) activity, and phagocytosis to investigate the immune traits of bacteria-challenged oriental armyworms, which were fed different P:C ratio diets. We found the oriental armyworms that were fed a 35:7 (P:C) diet showed higher phenoloxidase (PO) activity and stronger melanization, and those reared on a 28:14 (P:C) diet showed higher antimicrobial activity. However, different P:C diets had no apparent effect on the hemocyte number and phagocytosis. These results overall indicate that high P:C diets differently optimize humoral immune defense responses in oriental armyworms, i.e., PO-mediated melanization and antimicrobial peptide synthesis in response to bacteria challenge.

Two venom serpins from the parasitoid wasp Microplitis mediator inhibit the host prophenoloxidase activation and antimicrobial peptide synthesis.

Endoparasitoid wasps inject venom proteins into the hemocoel of host insects to ensure survival, growth, and development of their progenies by blocking host immunity. We previously identified ten serine protease inhibitors of the serpin superfamily in venom of the endoparasitoid wasp, Microplitis mediator, but it is unclear how these inhibitors may interact with host immune serine proteases. In this study, we investigated the functions of two serpins, MmvSPN-1 and MmvSPN-2, in the regulation of humoral immune responses in two hosts, the oriental armyworm Pseudaletia separate and the cotton bollworm Helicoverpa armigera, by dsRNA knockdown and biochemical assays using recombinant proteins. Knockdown of the two serpins resulted in increases in prophenoloxidase (PPO) activation and antimicrobial peptide (AMP) production in the hosts. After injection into the host hemocoel, the recombinant serpins inhibited PPO activation and AMP transcription. Mass spectrometry analysis of the pull-downs and in vitro reconstitution experiments revealed that HacSP29, a clip-domain serine protease in H. armigera, is the target of these two serpins. Therefore, these two inhibitors in the wasp venom may protect eggs from attacks by melanization and AMPs in the host insects.

Rho 1 participates in parasitoid wasp eggs maturation and host cellular immunity inhibition.

Endoparasitoid wasps introduce venom into their host insects during the egg-laying stage. Venom proteins play various roles in the host physiology, development, immunity, and behavior manipulation and regulation. In this study, we identified a venom protein, MmRho1, a small guanine nucleotide-binding protein derived from ovary in the endoparasitoid wasp Microplitis mediator and found that knockdown of its expression by RNA interference caused down-regulation of vitellogenin and juvenile hormone, egg production, and cocoons formation in the female wasps. We demonstrated that MmRho1 entered the cotton bollworm's (host) hemocytes and suppressed cellular immune responses after parasitism using immunofluorescence staining. Furthermore, wasp MmRho1 interacted with the cotton bollworm's actin cytoskeleton rearrangement regulator diaphanous by yeast 2-hybrid and glutathione s-transferase pull-down. In conclusion, this study indicates that MmRho1 plays dual roles in wasp development and the suppression of the host insect cellular immune responses.

Biosynthesis of Quercetin-Loaded Melanin Nanoparticles for Improved Antioxidant Activity, Photothermal Antimicrobial, and NIR/pH Dual-Responsive Drug Release.

Quercetin (QCT) is a promising dose-dependent nutraceutical that usually suffers from poor water solubility and low bioavailability issues. In this work, a novel QCT-loaded nanoscale delivery system was constructed based on the oxidative self-polymerization of melanin (Q@MNPs). The FT-IR, XRD, and Zeta potential analyses confirmed that QCT was successfully absorbed on the melanin nanoparticles (MNPs) via Π-Π and hydrogen bonding interactions. The encapsulation efficiency and particle size of Q@MNPs were 43.78% and 26.68 nm, respectively. Q@MNPs improved the thermal stability of QCT and the antioxidant properties in comparison to MNPs. Meanwhile, Q@MNPs presented fantastic photothermal conversion capacity and stability triggered by the NIR laser, which significantly enhanced the antibacterial capability with a sterilization rate of more than 98% against E. coli and S. aureus. More importantly, Q@MNPs exhibited NIR/pH dual-responsive drug release behavior and good biocompatibility (at concentrations of < 100 µg/mL). Thus, Q@MNPs show promising prospects for flavonoid delivery.

YTHDC2-Mediated circYTHDC2 N6-Methyladenosine Modification Promotes Vascular Smooth Muscle Cells Dysfunction Through Inhibiting Ten-Eleven Translocation 2.

Type 2 diabetes condition mediated vascular smooth muscle cell (VSMCs) dysfunction. However, the mechanism of VSMCs dysfunction in diabetic patients needs further elucidation. VSMCs are an important component of the vascular wall, participate in the process of vascular remodeling, and play a vital role in the vascular complications of diabetes. Studies have found that circular RNAs (circRNAs) play a key regulatory role in the occurrence and development of VSMCs dysfunction. In this study, we stimulated VSMCs with high glucose and identified a new circular RNA, circYTHDC2, using circRNA chip analysis. circYTHDC2 was highly expressed in VSMCs treated with high glucose. Knockout of circYTHDC2 significantly inhibited the proliferation and migration of VSMCs. Metformin treatment significantly inhibited the expression of YTHDC2 and circYTHDC2. The upstream mechanism analysis revealed that the stability of circYTHDC2 was regulated by YTHDC2-mediated m6A modification. Furthermore, circYTHDC2 negatively regulates the expression of Ten-Eleven Translocation 2 (TET2) by targeting the unstable motif of TET2 3'UTR, thereby promoting dedifferentiated "synthetic type" transformation of VSMC. Taken together, these results suggest that the YTHDC2/circYTHDC2/TET2 pathway is an important target of metformin in preventing the progression of VSMCs dysfunction under high glucose.

Scavenger receptor B1 mediates phagocytosis and the antimicrobial peptide pathway in the endoparasitic wasp Micropilits mediator.

Scavenger receptors (SRs) are a family of pattern recognition receptors (PRRs) in the immune system. They are required for phagocytosis and act as co-receptors of Toll-like receptors to regulate immune signaling pathways in the fight against pathogens. Little is known about the function of SRs in insects. Here, we reported on a member of the SR family from the parasitic wasp Micropilits mediator (designated MmSR-B1) that is responsive to bacterial infection. The recombinant extracellular CD36 domain of MmSR-B1 produced in Escherichia coli cells is capable of binding to peptidoglycans and bacterial cells, causing agglutination of bacteria. Furthermore, we demonstrated that double-stranded RNA-mediated knockdown of MmSR-B1 impedes hemocyte phagocytosis and downregulates the expression of antimicrobial peptide (AMP) genes defensins and hymenoptaecins. Knockdown of MmSR-B1 led to increased death of the wasps when challenged by bacteria. Our study suggests that MmSR-B1 mediates phagocytosis and the production of AMPs in M. mediator wasps.

Palladium-catalyzed bisthiolation of terminal alkynes for the assembly of diverse (Z)-1,2-bis(arylthio)alkene derivatives.

An efficient and straightforward palladium-catalyzed three-component cascade bisthiolation of terminal alkynes and arylhydrazines with sodium thiosulfate (Na2S2O3) as the sulfur source for the assembly of functionalized (Z)-1,2-bis(arylthio)alkene derivatives is described. Using 0.5 mol% IPr-Pd-Im-Cl2 as the catalyst, a wide range of terminal alkynes and arylhydrazines are well tolerated, thus producing the desired products in good yields with good functional group tolerance and excellent regioselectivity. Moreover, this protocol could be readily scaled up, showing potential applications in organic synthesis and material science.

The Effect of Waste Engine Oil and Waste Polyethylene on UV Aging Resistance of Asphalt.

Waste engine oil (WEO) and waste polyethylene (WPE) are two common wastes, which are easy to pollute the environment. As the primary material in road construction, natural asphalt is a non-renewable energy source and asphalt is vulnerable to ultraviolet (UV) radiation during the service life. It results in degradation of asphalt pavement performance. In this paper, 22 wt % to 82 wt % of WEO and WPE were used to modify asphalts and the UV aging simulation experiment was carried out. The physical parameters of asphalts before the UV aging experiment show that the asphalt containing 42 wt % WPE and 62 wt % WEO mixture (42 wt % WPE + 62 wt % WEO) has similar physical properties with that of the matrix asphalt. Besides, gel permeation chromatography (GPC) verifies that the molecular weight distribution of the asphalt containing 42 wt % WPE + 62 wt % WEO is close to that of the matrix asphalt. The storage stability test shows that 42 wt % WPE + 62 wt % WEO has good compatibility with the matrix asphalt. The functional groups and micro-morphology of asphalts before and after the UV aging experiment were investigated by Fourier transform infrared spectroscopy (FTIR) and atomic force microscopy (AFM). FTIR results display that 42 wt % WPE + 62 wt % WEO can effectively reduce the formation of carbonyl and sulfoxide functional groups. AFM shows that 42 wt % WPE + 62 wt % WEO can also retard the formation of a "bee-like" structure in asphalt after the UV aging experiment. Based on the above results, it can be concluded that WEO and WPE mixture can replace part of asphalt and improve the UV aging resistance of asphalt.

Manipulation of the silkworm immune system by a metalloprotease from the pathogenic bacterium Pseudomonas aeruginosa.

Antimicrobial peptide (AMP) production and melanization are two key humoral immune responses in insects. Induced synthesis of AMPs results from Toll and IMD signal transduction whereas melanization depends on prophenoloxidase (PPO) activation system. During invasion, pathogens produce toxins and other virulent factors to counteract host immune responses. Here we show that the pathways leading to PPO activation and AMP synthesis in the silkworm Bombyx mori are affected by a metalloprotease, named elastase B, secreted by Pseudomonas aeruginosa (PAO1). The metalloprotease gene (lasB) was expressed shortly after PAO1 cells had been injected into the larval silkworm hemocoel, leading to an increase of elastase activity. Injection of the purified PAO1 elastase B into silkworm hemolymph compromised PPO activation. In contrast, the protease caused a level increase of gloverin, an AMP in the hemolymph. To verify our results obtained using the purified elastase B, we infected B. mori with PAO1 ΔlasB mutant and found that PO activity in hemolymph of the PAO1 ΔlasB-infected larvae was significantly higher than that in the wild type-infected. The mutant-inhabited hemolymph had lower levels of gloverin and antimicrobial activity. PAO1 ΔlasB showed a decreased viability in the silkworm hemolymph whereas the host had a lower mortality. In addition, the effects caused by the ΔlasB mutant were restored by a complementary strain. These data collectively indicated that the elastase B produced by PAO1 is an important virulent factor that manipulates the silkworm immune system during infection.

Effect of ultrasonic field on the enzyme activities and ion balance of potential pathogen Saccharomyces cerevisiae.

OBJECTIVES: This study aimed at investigating the enzyme activities and ion concentrations in potential pathogen S.cerevisiae upon ultrasonic treatment. METHODS: The activities of ATPase and antioxidase were identified by ATPase, SOD, and CAT assay kits following the instructions. Extracellular Ca2+ and K+ concentrations were determined in an atomic absorption spectrometer with calcium and potassium hollow-cathode lamps as radiation sources. RESULTS: SOD and CAT activities were enhanced by relatively low ultrasonic power at early time points and reduced to lower levels. Total ATPase, Na+/K+-ATPase, and Ca2+/Mg2+-ATPase activities were reduced by ultrasonic field, with higher reducing rate at stronger ultrasonic power and early time points. In addition, ultrasonic field disturbed the Ca2+ and K+ balances in S.cerevisiae cells. CONCLUSIONS: Ultrasonic field resulted in the reduce even the lost of S.cerevisiae cell viability.

Effect of ultrasound treatment conditions on Saccharomyces cerevisiae by response surface methodology.

OBJECTIVES: This study aimed to investigate the effect of different ultrasound treatment conditions on the inactivation of Saccharomyces cerevisiae with the application of response surface methodology (RSM). METHODS: Ultrasound treatment were applied on different concentrations of S. cerevisiae cells with different pH, temperature, ultrasound power, irradiating time, and pulse duty ratio. Cell viability was determined by plate counting method. Response surface methodology was used to analyze the correlation among various factors. RESULTS: Limited with low ultrasound power, lower pH value slightly improved the ultrasound treatment efficiency. Also, higher nonlethal temperature and ultrasound power, longer irradiation time, and lower pulse duty ratio facilitated the inactivation of S. cerevisiae. Cell concentration had no effect on ultrasound efficiency. CONCLUSIONS: Ultrasound power played the most important role in the ultrasound irradiation process according to RSM analyses. Information derived from this study may aid in the control of the sublethal injury of S. cerevisiae during ultrasound treatment in food industry.

Transcriptome and expression profiling analysis of the hemocytes reveals a large number of immune-related genes in mud crab Scylla paramamosain during Vibrio parahaemolyticus infection.

BACKGROUND: Mud crab Scylla paramamosain is an economically important marine species in China. However, frequent outbreaks of infectious diseases caused by marine bacteria, such as Vibrio parahaemolyticus, result in great economic losses. METHODOLOGY/PRINCIPAL FINDINGS: Comparative de novo transcriptome analysis of S. paramamosain infected with V. parahaemolyticus was carried out to investigate the molecular mechanisms underlying the immune response to pathogenic bacteria by using the Illumina paired-end sequencing platform. A total of 52,934,042 clean reads from the hemocytes of V. parahaemolyticus-infected mud crabs and controls were obtained and assembled into 186,193 contigs. 59,120 unigenes were identified from 81,709 consensus sequences of mud crabs and 48,934 unigenes were matched proteins in the Nr or Swissprot databases. Among these, 10,566 unigenes belong to 3 categories of Gene Ontology, 25,349 to 30 categories of KEGG, and 15,191 to 25 categories of COG database, covering almost all functional categories. By using the Solexa/Illumina's DGE platform, 1213 differentially expressed genes (P<0.05), including 538 significantly up-regulated and 675 down-regulated, were detected in V. parahaemolyticus-infected crabs as compared to that in the controls. Transcript levels of randomly-chosen genes were further measured by quantitative real-time PCR to confirm the expression profiles. Many differentially expressed genes are involved in various immune processes, including stimulation of the Toll pathway, Immune Deficiency (IMD) pathway, Ras-regulated endocytosis, and proPO-activating system. CONCLUSIONS/SIGNIFICANCE: Analysis of the expression profile of crabs under infection provides invaluable new data for biological research in S. paramamosain, such as the identification of novel genes in the hemocytes during V. parahaemolyticus infection. These results will facilitate our comprehensive understanding of the mechanisms involved in the immune response to bacterial infection and will be helpful for diseases prevention in crab aquaculture.

Prospect theory based estimation of drivers' risk attitudes in route choice behaviors.

This paper applied prospect theory (PT) to describe drivers' route choice behavior under Variable Message Sign (VMS), which presented visual traffic information to assist them to make route choice decisions. A quite rich empirical data from questionnaire and field spot was used to estimate parameters of PT. In order to make the parameters more realistic with drivers' attitudes, they were classified into different types by significant factors influencing their behaviors. Based on the travel time distribution of alternative routes and route choice results from questionnaire, the parameterized value function of each category was figured out, which represented drivers' risk attitudes and choice characteristics. The empirical verification showed that the estimates were acceptable and effective. The result showed drivers' risk attitudes and route choice characteristics could be captured by PT under real-time information shown on VMS. For practical application, once drivers' route choice characteristics and parameters were identified, their route choice behavior under different road conditions could be predicted accurately, which was the basis of traffic guidance measures formulation and implementation for targeted traffic management. Moreover, the heterogeneous risk attitudes among drivers should be considered when releasing traffic information and regulating traffic flow.

Characterization of microRNAs in mud crab Scylla paramamosain under Vibrio parahaemolyticus infection.

BACKGROUND: Infection of bacterial Vibrio parahaemolyticus is common in mud crab farms. However, the mechanisms of the crab's response to pathogenic V. parahaemolyticus infection are not fully understood. MicroRNAs (miRNAs) are a class of small noncoding RNAs that function as regulators of gene expression and play essential roles in various biological processes. To understand the underlying mechanisms of the molecular immune response of the crab to the pathogens, high-throughput Illumina/Solexa deep sequencing technology was used to investigate the expression profiles of miRNAs in S. paramamosain under V. parahaemolyticus infection. METHODOLOGY/PRINCIPAL FINDINGS: Two mixed RNA pools of 7 tissues (intestine, heart, liver, gill, brain, muscle and blood) were obtained from V. parahaemolyticus infected crabs and the control groups, respectively. By aligning the sequencing data with known miRNAs, we characterized 421 miRNA families, and 133 conserved miRNA families in mud crab S. paramamosain were either identical or very similar to existing miRNAs in miRBase. Stem-loop qRT-PCRs were used to scan the expression levels of four randomly chosen differentially expressed miRNAs and tissue distribution. Eight novel potential miRNAs were confirmed by qRT-PCR analysis and the precursors of these novel miRNAs were verified by PCR amplification, cloning and sequencing in S. paramamosain. 161 miRNAs (106 of which up-regulated and 55 down-regulated) were significantly differentially expressed during the challenge and the potential targets of these differentially expressed miRNAs were predicted. Furthermore, we demonstrated evolutionary conservation of mud crab miRNAs in the animal evolution process. CONCLUSIONS/SIGNIFICANCE: In this study, a large number of miRNAs were identified in S. paramamosain when challenged with V. parahaemolyticus, some of which were differentially expressed. The results show that miRNAs might play some important roles in regulating gene expression in mud crab under V. parahaemolyticus infection, providing a basis for further investigation of miRNA-modulating networks in innate immunity of mud crab.

Synthesis and antitumor activity of novel nitrogen mustard-linked chalcones.

A series of nitrogen mustard-linked chalcones were synthesized and evaluated for their antitumor activity in vitro against the K562 and HepG2 cell lines. The aldol condensation of [N,N-bis(chloroethyl)-3-amino]-acetophenone (2) with aromatic aldehydes afforded the nitrogen mustard-linked chalcones. Among the analogs tested, compounds 5e and 5k exhibited significant anti-proliferation activities against K562 cells with IC50 values of 2.55 and 0.61 µM, respectively, which revealed higher cell toxicity than the standard drugs cisplatin (IC50>200 µM) and adriamycin (IC50=14.88 µM). The methoxyl and N,N-dimethyl groups on the B-ring of the chalcone frame enhanced the inhibitory activities against both the K562 and HepG2 cell lines. The structure-activity relationship study indicated that the inhibitory activities significantly varied with the position(s) and species of the substituted group(s).