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Mitogen-activated protein kinase kinase kinase 4 (MAP3K4) is a multifunctional mediator of the conserved MAPK signaling pathway that plays essential roles in the regulation of immune responses in mammals. However, the function of teleost MAP3K4s in innate immunity, especially in the intestinal immune system, is still poorly understood. In the current study, we identified a fish MAP3K4 homolog (CiMAP3K4) in Ctenopharyngodon idella as well as its immune function in intestine following bacterial infection in vivo and in vitro. The open reading frame (ORF) of CiMAP3K4 encodes putative peptide of 1544 amino acids containing a predicted serine/threonine protein kinase (S_TKc) domain with high identity with other fish MAP3K4s. Phylogenetic analysis revealed the CiMAP3K4 belonged to the fish cluster and showed the closest relationship to Pimephales promelas. Quantitative real-time PCR (qRT-PCR) analysis revealed that CiMAP3K4 transcripts were widely distributed in all tested tissues, especially with high expression in the muscle and intestine of healthy grass carp. In vitro, CiMAP3K4 gene expression was upregulated by bacterial PAMPs (lipolysaccharide (LPS), peptidoglycan (PGN), L-Ala-γ-D-Glu-meso-diaminopimelic acid (Tri-DAP) and muramyl dipeptide (MDP)) and pathogens (Aeromonas hydrophila and Aeromonas veronii) in primary intestinal cells. In vivo, the mRNA expression levels of CiMAP3K4 in the intestine were significantly induced by bacterial MDP challenge in a time-dependent manner; however, this effect could be inhibited by the bioactive dipeptides β-alanyl- l -histidine (carnosine) and alanyl-glutamine (Ala-Gln). Moreover, CiMAP3K4 was located primarily in the cytoplasm, and its overexpression increased the transcriptional activity of AP-1 in HEK293T cells. Collectively, these results suggested that CiMAP3K4 might play an important role in the intestinal immune response to bacterial infections, which paves the way for a better understanding of the intestinal immune system of grass carp.

In this study, we aimed to develop B. subtilis spore coat protein A (CotA) for the enzymatic determination of bilirubin. Firstly, molecular docking and oxidation kinetic analysis confirmed the feasibility of CotA for oxidizing bilirubin. Secondly, CotA showed pH-preferable oxidization performance to direct bilirubin (DB) in acidic conditions and an alkaline-catalytic oxidation capacity to total bilirubin (TB). Mechanism analysis results confirm that the conformational changes of CotA, DB and UB caused by pH changes are responsible for the selective oxidation of DB and TB by CotA. Then, CotA exhibits better structural characteristics and enzymatic performance than M. verrucaria-derived bilirubin oxidase (Mv-BOD). Besides, the strong anti-interference ability helps CotA adapt to complex catalytic environment in the detection of DB and TB. Our results prove that CotA can be used as a promising candidate bio-enzymatic detection reagent for DB and TB.

We report the self-assembly of an ultra-pH-sensitive hydrogel from a simple dipeptide derivative, ferrocene-diphenylalanine (noted as Fc-FF). The pure Fc-L-Phe-L-Phe-OH (noted as (L)-Fc-FF) or Fc-D-Phe-D-Phe-OH (noted as (D)-Fc-FF) peptide could self-assemble into right- or left-handed nanohelices in a mixture of water and organic solvent leading to formation of stable chiral hydrogels at a very narrow pH range of 5.7-5.9. Intriguingly, if we used a racemic mixture (noted as (L,D)-Fc-FF) of (L)-Fc-FF and (D)-Fc-FF, stable hydrogels could only be formed at much lower pH values with a weak mechanical property. The results indicated that the molecular chirality has great influence on the gelation of the peptides. Moreover, we investigated the gelation behaviour of the peptides in various solvents. Here the water molecules are essential in directing the chiral self-assembly of Fc-FF into entangled chiral nanostructures, leading to the formation of stimuli-responsive hydrogels. And in the presence of water molecules, a small amount of organic co-solvents also has significant effect on the gelation of Fc-FF peptides. This work demonstrated that the gelation of such a bioorganometallic molecule is very sensitive to the molecular conformation of itself and the self-assembly environment (pH, solvents, etc.), which has potential applications in chemical sensing and biomedicine.

Zinc oxide (ZnO) nanomaterials are functional photocatalysts that show excellent photocatalytic activity, among which diverse morphologies, such as nanospheres, nanorods, nanoleaves, nanodisks, or nanoflowers, often possess different photocatalytic activities. In this study, we employed a simple ultrasonic treatment to controllably synthesize three types of three-dimensional (3D) fluffy ZnO nanoflowers with different nanostructures. Specifically, one nanoflower with smooth edges was formed via direct hydrothermal reaction without ultrasonic treatment (denoted as ZnO-0), and the other two with different jagged margins were obtained by ultrasonic treatment at 250 W and 950 W, respectively, before the hydrothermal procedure (denoted as ZnO-250 and ZnO-950). The results showed that the size, specific surface area, crystallite dimension, intrinsic donor defects and the signals of reactive radicals of ZnO nanoflowers all decreased with increasing ultrasonic intensity. Furthermore, the photocatalytic performance of such ZnO nanoflowers with similar morphologies but different nanostructures was investigated using methyl orange (MO) as a model pollutant. As expected, ZnO-0 possessed the highest photocatalytic activity with a kinetic constant of 0.0478 min−1, while the value decreased to 0.0251 min−1 and 0.013 min−1 for ZnO-250 and ZnO-950 under simulated sunlight irradiation, respectively, suggesting that the photocatalytic efficiency of ZnO nanoflowers decreased due to the ultrasonic treatment. These findings provide structural insight into the photocatalytic activity of ZnO nanoflowers.

The p38 mitogen-activated protein kinases (MAPKs) are essential cytoplasmic signal molecules of innate immune pathways that play a vital role in host immune defense responses to pathogenic challenges. In this study, two fish p38 genes (Cip38α and Cip38β) were characterized for the first time from the grass carp Ctenopharyngodon idella. Similar to other reported p38MAPKs, both Cip38α and Cip38β contained a conserved phosphorylation motif (Thr-Gly-Tyr, TGY) and a substrate binding site (Ala-Thr-Arg-Trp, ATRW) in the serine/threonine protein kinase (S_TKc) domain. Expression profile analysis showed that Cip38α and Cip38β mRNAs were broadly expressed in all of the examined tissues and developmental stages of C. idella. In addition, in vivo injection experiments directly revealed that Cip38α and Cip38β showed strong responsiveness to Aeromonas hydrophila and muramyl dipeptide (MDP) challenges, and their expression levels were significantly upregulated in the intestine of grass carp. Additionally, the MDP-induced expression levels of intestinal inflammatory cytokines (TNF-α and IL-15) and an antimicrobial peptide (β-defensin) were significantly inhibited by the p38MAPK-specific inhibitor SB203580. Moreover, the nutritional dipeptide carnosine and Ala-Gln were found to significantly suppress the bacterial MDP-induced expression of p38MAPK pathway genes and inflammatory cytokines in the intestine of grass carp. Finally, overexpression analysis demonstrated that Cip38α and Cip38β could act as efficient activators in the regulation of AP-1 signaling pathways through interaction with CiMKK6. Altogether, this study provided experimental evidence of the presence of a functional p38 pathway in grass carp, which revealed its involvement in the intestinal immune response to bacterial challenges in bony fish.

In order to popularize magnesium oxysulfate (MOS) cement in civil engineering, it is very necessary to investigate its carbonation related durability. It was found that carbonization can increase compressive and flexural strength of MOS cement to some extent and compressive strength of MOS cement increases gradually at first, then decreases during carbonation process. The displacement to ultimate compressive force of hardened MOS cement paste carbonated for 28 days is larger than that of non-carbonated one at the same age. Evaporable water testing and X-ray computed tomography (X-CT) were adopted to examine the porous structure of MOS cement paste with and without carbonation. It was found that carbonation refines the meso-pore (≧38 µm meters) volume ratio of hardened MOS cement with 0.5 water-to-cement ratio by about 17%. X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) were adopted to analyze the carbonation characteristics of MOS cement. The results indicate that magnesium hydroxide and some basic magnesium oxysulfate phase are obviously neutralized with carbon dioxide to form magnesium carbonate phases in the hardened MOS cement, which leads to the refined porous structures and corresponding increased toughness.

Nanozymes, defined as nanomaterials with enzyme-like activity, have attracted extensive interest in both fundamental and applied research. Laccases are members of the multi-copper oxidases, which are utilized as green catalysts in the environmental catalysis and biochemical industry. In this paper, we report a facile strategy for the preparation of a new class of nanozyme (denoted as CH-Cu) with laccase-like activity inspired by the structure of the active site and the electron transfer pathway of laccase via the coordination of Cu+/Cu2+ with a cysteine (Cys)-histidine (His) dipeptide. The CH-Cu nanozymes exhibit excellent catalytic activity, recyclability and substrate universality and have a similar Km (Michaelis constant) and a higher vmax (maximum rate) than laccase at the same mass concentration. They are robust under a variety of conditions, such as extreme pH, high temperature, long-term storage and high salinity, which can cause severe loss in the catalytic activity of laccase. Higher efficacy of the CH-Cu nanozymes compared with laccase in the degradation of chlorophenols and bisphenols is also demonstrated in a batch reaction. Furthermore, a method for the quantitative detection of epinephrine by a smart phone is established based on the CH-Cu nanozymes. We believe that this nanozyme has promising applications in environmental catalysis and rapid detection and expect that combining key peptides as metal ligands with metal ions to mimic the structure of the catalytic center of a natural enzyme will be a general and important strategy for the design and synthesis of a new type of nanozyme that can be used in various applications.

Alcohol intake increases the risk of cancer development. Approximately 3.6% human cancers worldwide derive from chronic alcohol drinking, including oral, liver, breast and other organs. Our studies in vivo and in vitro have demonstrated that diluted ethanol increase RNA Pol III gene transcription and promotes cell proliferation and transformation, as well as tumor formation. However, it is unclear about the effect of red wines on the human cancer cells. In present study, we investigated the roles of red wine in human cancer cell growth, colony formation and RNA Pol III gene transcription. Low concentration (12.5 mM to 25 mM) of ethanol enhances cell proliferation of breast and esophageal cancer lines, whereas its higher concentration (100 mM to 200 mM) slightly decreases the rates. In contrast, red wines significantly repress cell proliferation of different human cancer lines from low dose to high dose. The results reveal that the red wine also inhibits colony formation of human breast cancer and esophageal carcinoma cells. The effects of repression on different human cancer lines are in a dose-dependent manner. Further analysis indicates that ethanol increases RNA Pol III gene transcription, whereas the red wines significantly reduce transcription of the genes. Interestingly, the effects of mature wine (brick red) on cancer cell phenotypes are much stronger than young wine (intense violet). Together, these new findings suggest that red wines may contain some bioactive components, which are able to inhibit human cancer cell growth and colony formation.

5-Formyl-2-furancarboxylic acid (FFCA) is increasingly important building blocks, which has promising applications in fuels, chemical intermediates, drugs, etc. However, highly selective oxidation of 5-hydroxymethylfurfural (5-HMF) to FFCA by enzyme catalysis is a new challenging problem due to the absence of specific enzymes. Recently we identified a kind of laccase (CotA-TJ102) from Bacillus subtilis TJ-102 could convert selectively 5-HMF to FFCA, and no further oxidation production (such as FDCA). The initial conditions resulted in low 5-HMF conversion ratio at 23.5% and low FFCA yield at 19.3% after 96 h, but high FFCA selectivity at 82.4%. Then the proposed mechanism in detail for the selective oxidation of HMF to FFCA by CotA-TJ102 was deduced and discussed. After optimization of reaction parameters such as pH, TEMPO concentration and temperature, FFCA could be obtained in a high yield of 98.55% with a 5-HMF conversion ratio of nearly 100% after a short reaction time of 12 h. Finally, after immobilization of CotA-TJ102 on magnetic nanoparticles, the FFCA yield remained 83.28% for 10 of recycling times based on the high FFCA selectivity (>96%), which reflected high stability and good reusability. Therefore, this enzymatic approach constitutes a promising method for the green production of FFCA.

2,5-Diformylfuran (2,5-DFF) is an important furan-based chemical with broad application potential. To address the difficulties in isolating/purifying the product and recycling the 2,2,6,6- tetramethylpiperidine-1-oxy (TEMPO)-based catalysts, we employed a solvent with a low boiling point in which TEMPO has a low solubility to investigate the effects of TEMPO and its analogs on catalyzing the oxidation of 5-hydroxymethylfurfural (5-HMF). The results indicated that in ethyl acetate with the substitution of one H of TEMPO at the C4 position with either an acetamido ( NH-Ac), methoxy ( OCH3), hydroxy ( OH) or isothiocyanato ( N C S) group, the resulting compound had a product selectivity similar to that of TEMPO (>86%), except for the 4-isothiocyanato-TEMPO-catalyzed reaction. However, after two Hs were substituted with an oxo group ( O), the catalytic efficiency significantly decreased (product yield 16%, selectivity 52%). Based on both the catalytic efficiency and catalyst solubility, a highly efficient method for preparing 2,5-DFF was established using ethyl acetate as the solvent, 4-acetamido-TEMPO as the catalyst and Fe(NO3)3 and NaCl as cocatalysts. At 40 °C with a substrate concentration of 0.125 mol/L, 5-HMF was completely converted in 2 h, and the yield of 2,5-DFF was 89%. Moreover, when the substrate concentration increased to 0.25 mol/L, the yield of 2,5-DFF was 87%. When 4-acetamido-TEMPO was recovered by centrifugation, the product yield remained as high as 86% after three cycles, demonstrating good recyclability.

Cu 2+ plays a decisive role for the bio-oxidation in the active center of laccase. In the fermentation-purified process, the loss of Cu 2+ reduces the activity and the high cost limits the application of laccase. In this study, we developed a fermentation-permeabilization combined process based on the regulation of Cu 2+ binding time to produce the permeabilized cells, in which Cu 2+ can enter the cells freely to greatly improve the laccase activity and reduce the immobilization cost by about 19 times. So the permeabilized-cells is suitable for biodegradation of antibiotic pollution in the environment, which was applied for the biodegradation of ciprofloxacin (CIP) and tetracycline-HCl (TCH) and the degradation efficiency reached 95.42% and 98.73%, respectively, with low ecotoxicity of the degradation products. Finally, we theoretically analyzed the degradation mechanism and the degradation pathways. Therefore, this study provided a low-cost, eco-friendly, and widely applicable method for organic pollutants removal.

In this study, using furfuryl alcohol as the precursor carbon and mesoporous silica as the template, and furfuryl alcohol-derived mesoporous carbon (FMC) was prepared. The specific surface area of FMC was 1022.61 m2/g, the pore volume was 1.71 cm3/g, and the mesoporous volume was 98.8%. Based on the adsorption kinetics of pharmaceuticals onto the FMC in synthetic urine, equilibrium adsorption was reached in 120 min, and it followed a pseudo-second-order model. The adsorption isotherms were well-fitted by the Sips isotherm model, and the saturated adsorption capacities of diclofenac and oxytetracycline in fresh urine were 411.8 mg/g and 465.9 mg/g, respectively. Batch experiment results showed that pharmaceutical removal was strongly influenced by urine components such as sodium chloride, urea, and ammonium hydroxide. The adsorption of diclofenac and oxytetracycline was influenced by many factors including π-π interactions, hydrogen bonds, and electrostatic forces. FMC exhibited excellent reusability and retained urine nutrients during pharmaceutical adsorption.

An alkaline-modified natural sponge was applied as support of gold and silver nanocatalysts and a deformable reactor to hold the reaction solution, quickly adsorb several kinds of environmental pollutants (4-nitrophenol, hexavalent chromium, and methyl orange) and efficiently catalyse their conversion. As the reaction solution is sustained in the connecting micro-pores constructed by the stretched and knotted sponge fibers, the catalytic reaction is free from external mass transfer and suffers slight internal mass transfer limitation. This batch-wise reaction system employing an operation procedure of injection-extrusion achieved a nearly complete conversion of the reactants within 1 min or much less. This compressible support can also be packed in a continuous flow reaction system and can be repeated use for 37 times without any loss of catalytic activity. This study provides a generalizable and straightforward strategy for utilizing deformable soft supports that may inspire some unique industrial applications.

A rational design method to improve β-mannanase (ManTJ102) thermostability was developed successfully in this study. The flexible area of residues 330–340 in ManTJ102 was firstly selected from analysis of molecular dynamics simulation and then the critical amino acid residue (Ala336Pro) with the lowest mutation energy was determined by virtual mutation, whose mutant was named as Mutant336. Afterward, the dynamics transition temperature (Tdtt) of ManTJ102 and Mutant336 was evaluated by simulated annealing and heating, and Mutant336 with higher Tdtt was implemented for experimental verification of the enzyme thermostability. As a result, the half-life of Mutant336 activity was 120 min at 60 °C, which was 24-fold of ManTJ102, and the irreversible thermal denaturation constant of Mutant336 was only about 2/5 of ManTJ102, indicating that Mutant336 has better thermostability than ManTJ102. Furthermore, Mutant336 has much higher β-mannanase activity and specific activity than ManTJ102. Therefore, Mutant336 was more suitable to further research for applications.

Nanoscale zero valent iron (nZVI) and bimetallic nZVI has been widely applied to remove various contaminants from water and soil, but their activities can be greatly reduced due to the precipitation of the generated Fe ions. Here we adopted tannic acid (TA) as stabilizer to synthesize high activity Fe-Pd nanoparticles (NPs) and scavenger of the generated Fe ions to keep the rapid dechlorination of two organochlorides. Inductively coupled plasma optical emission spectrometry and UV–vis spectroscopy measurements indicated the generated Fe ions were harvested by TA and formed bis- and tris-TA-iron complexes in the dechlorination solution. TA promoted 40-fold and 10-fold increase in dechlorination efficiency of p-chlorophenol and florfenicol attributing to the complexation of Fe ion by TA. This strategy may inspire novel ways to deal with the passivation of nZVI caused by the precipitation of Fe ions and benefits for the developing of longevity and high-efficient nZVI materials.

5-Hydroxymethylfurfural is an important bio-based chemical that shows broad application potential. In the present study, the use of chitosan nanoparticles was proposed to adsorb and recycle chromium ions from aqueous solution with Cr(III) and Cr(VI) removal rates of up to 97.5% and 81.2%, respectively. A chromium-based ceramic powder catalyst was designed and synthesized, the structural properties of the catalyst were characterized, and the catalytic performances of the catalyst were evaluated. The results showed that after chromium was doped into the blank ceramic, the main lattice structure of the ceramic was preserved. Both the specific surface area and pore diameter decreased, and a portion of the Cr(VI) was transformed to Cr(III). The chromium-based ceramic powder and sulfuric acid co-catalyzed the one-pot synthesis of 5-hydroxymethylfurfural from glucose in the water/dimethyl sulfoxide system, with the product yield reaching 64.7%. After the catalyst was reused 5 times, the product yield was still close to 60%. Thus, the catalyst exhibited good recyclability. The results of this study provide a good example of the one-step synthesis of 5-hydroxymethylfurfural from glucose.

5-Hydroxymethylfurfural (HMF), converted from sustainable biomass resources, is a versatile intermediate in chemical industry. A kind of ceramic powder containing Sn4+ ions (Sn-CP) with good hardness was synthesized by a rapid method for the isomerization of glucose. HCl was introduced for catalysing the fructose-to-HMF reaction and promoting the isomerization reaction. Production of HMF from glucose catalysed by Sn-CP/HCl demonstrated high HMF yield (63.9%) and effective catalyst recyclability (HMF yield >53.7%). Moreover, when fructose and poly-saccharides (sucrose, inulin, starch and maltose) as feedstocks, HMF yields of more than 55% were obtained. Hence integration of catalytic reaction and conversion of carbohydrates to HMF provided a green process over inexpensive catalyst.

Currently, the traditional fermentation using konjac powder for β-mannanase production presents operational difficulties and high energy consumption, because of the individual hot air sterilization for konjac powder. A simply enzymatic hydrolysis pretreatment for konjac powder was developed to solve the problems of the traditional process in a 7-L fermenter. In the new process, when hydrolysis yield of konjac powder was above 50%, the media became liquid state from gelatinous state and could be sterilized immediately, avoiding the hot air sterilization and solving the operational difficulties. Interestingly, the new process didn’t have negative influence on β-mannanase production. Additionally, it could save close to 23% of power consumption during the whole fermentation. For another example, it did work well using locust bean gum for β-mannanase production in a 7-L fermenter. Therefore, the new process might be scaled up for industrial production using mannan-based bioresource as substrate.

Chirality is one of the most important structural features in biological systems. Peptides, as a class of small biomolecules, can self-assemble into various chiral supramolecular polymers with unprecedented levels of diversity and complexity, which have practical ramifications in various fields from catalysis to sensing, optics, and biomedicine. In this review, we will focus on the chiral transfer during peptide self-assembly from the molecular scale to the nano-, micro- and mesoscale, and highlight the vital roles of chirality in determining the structures and functions of the self-assembled supramolecular polymeric materials. By rational control of the chiral self-assembly of peptide monomers, supramolecular polymers with sophisticated hierarchical structures and enhanced chemical and biological functions can readily be achieved. These can provide inspiration for researchers to design highly efficient asymmetric catalysts, templates fabricating materials with tunable chiroptics, or to synthesize biomaterials with chirality-encoded properties for biomedical applications.

Profiting from Pt-like electronic structure and high electrical conductivity, molybdenum phosphide (MoP) has received widespread attention as a hydrogen evolution reaction (HER) catalyst. Although various effective strategies have been developed to regulate the bare MoP synthesis, the electrocatalytic performance of MoP is still far from satisfactory, especially in pH-universal applications. Recently, doping of heterogeneous elements, particularly for rare earth (RE) metals, has been emerged as an effective method to precisely regulate the local electronic structure of phosphides. Herein, the novel and controllable La/Yb-doped MoP nanoparticles encapsulated in nitrogen-doped carbon matrix (MoP@NC) are synthesized. The as-prepared La/Yb-doped catalysts (RE-MoP@NC (RE = La, Yb)) exhibit fantastic HER electrocatalytic performance for both activity and durability in a wide pH range. Detailed structural characterizations and density functional theory (DFT) calculations validate that the electronic densities around Mo and P atoms are effectively tuned by La/Yb doping atoms, resulting in the optimization in Gibbs free energy of MoP toward the hydrogen adsorption, which can boost its intrinsic HER activity. This research enriches RE-modified catalysts and reveals that the RE metal doping can be extended to other non-noble metal catalysts as a general method.

In recent years, nanocellulose-based materials have been increasingly applied as a lot of biosorbents for the treatment of water pollutants due to their large specific surface area, easy modification, environmental friendliness, and reproducibility. In this review, surface modification of nanocellulose-based adsorbents with various effective adsorption groups is described, as well as polymer grafting and hybrid composite fabrication. The adsorption mechanisms involved in the adsorption process of pollutants by adsorbents are further analysed and summarized. The regeneration methods of nanocellulose adsorbents with different adsorption mechanisms are also demonstrated. In addition, this paper also briefly describes the forms of nanocellulose-based adsorbents with large-scale application including membranes, gels, flocculants and magnetic composites.

Black phosphorus (BP) nanodots inevitably interact with electrolyte ions because of their extensive applications in many fields. In biomedical applications, BP-nanodots might encounter various salt ions in body fluids and have a propensity to aggregate, further resulting in an enhanced toxicity risk. In this study, the effects of electrolyte cations (Na+, K+, Ca2+, and Mg2+) on the aggregation kinetics of BP-nanodots were investigated by means of dynamic light scattering (DLS). The critical coagulation concentration values of BP-nanodots were determined to be 242.11 mM NaCl, 95.01 mM KCl, 1.92 mM CaCl2, and 5.88 mM MgCl2. Divalent cations accelerated aggregation of BP-nanodots because of the stronger surface charge screening and their bridging with oxidized phosphorus species (PxOy) available on BP-nanodot surfaces. Furthermore, the divalent cations may interact with PxOy to form insoluble complex coatings on the nanoparticle surface and shield the charge. After modification with PEG, BP-nanodots presented enhanced stability in the presence of 2.5 mM CaCl2. This work is of great significance for understanding the aggregation behaviors of ultrasmall nanoparticles such as BP-nanodots in electrolyte solutions and provides insight into their safety assessment associated with biomedical applications.

Peptide assembly provides a new approach for the construction of enzyme-like catalysts. However, low stability is still a big challenge for the peptide-based enzyme mimics because the non-covalently driven structures are often destroyed when exposed to environmental changes. Inspired by the catalytic centre of natural horseradish peroxidase (HRP) and the biological function of dityrosine in structural proteins, in this study, we presented the construction of a stable peptide-based HRP mimic (CCA-YH) through introducing covalent bonds. CCA-YH was obtained through the co-assembly of YYHYY and hemin, followed by a ruthenium-mediated photo-crosslinking reaction. Catalytic activity assay demonstrated that CCA-YH displayed enhanced catalytic ability than the non-covalent co-assemblies (CA-YH). Remarkably, CCA-YH exhibited superior long-term stability than CA-YH and natural HRP under different pH, temperature, ion strength and organic medium environments, as well as a favorable reusability. Finally, CCA-YH was applied to the colorimetric detection of uric acid and sarcosine in human urine, which displayed satisfactory sensitivity, specificity and reproducibility. This is the first report the construction of stable peptide-based enzyme mimic based on covalent co-assembly strategy.

Whey powder, a by-product of dairy industry, is an attractive raw material for value-added products. In this study, utilization of whey powder as substrate for low-cost preparation of β-galactosidase as main product and ethanol as by-product were investigated by a litre-scale integrated strategy, encompassing fermentation, isolation, permeabilization and spray drying. Firstly, through development of low-cost industrial culture and fed-batch strategies by Kluyveromyces lactis, 119.30U/mL β-galactosidase activity and 16.96mg/mL by-product ethanol were achieved. Afterward, an up-dated mathematic model for the recycling permeabilization was established successfully and 30.4g cells sediment isolated from 5L fermentation broth were permeabilized completely by distilled ethanol from broth supernatant. Then β-galactosidase product with 5.15U/mg from protection of gum acacia by spray drying was obtained. Furthermore, by-product ethanol with 31.08% (v/v) was achieved after permeabilization. Therefore, the integrated strategy using whey powder as substrate is a feasible candidate for industrial-scale implementation.

Studies in several cancers have suggested that miR-218 has anti-tumor activities, but its function is yet to be elucidated. In this study, we investigated the regulation and function of miR-218 (miR-218-5p) in the cell cycle progression of gastric cancer (GC). We found that miR-218 could suppress proliferation of gastric cancer cells, induce cell cycle arrest at the G1 phase and inhibit tumor growth and metastasis in vivo. We also demonstrated that miR-218 specifically targeted the 3′-UTR regions of CDK6 and cyclin D1 and inhibited the expression of these molecules, which in turn repressed the pRb/E2F1 signaling pathway. Overexpression of CDK6 and Cyclin D1 reversed miR-218-mediated inhibition of pRB/E2F1 signaling and attenuated the miR-218-induced cell cycle arrest. More importantly, miR-218 expression was significantly reduced and inversely correlated with the levels of CDK6 and Cyclin D1 in gastric cancer tissues. Decreased miR-218 expression was also correlated with advanced clinical stage, lymph node metastasis, and poor prognosis in gastric cancer patients. Furthermore, we showed that miR-218 expression was directly activated by E2F1 through the transactivation of miR-218 host genes, SLIT2 and SLIT3, revealing a negative feedback regulation of miR-218 expression. Taken together, our results describe a regulatory loop miR-218-CDK6/CyclinD1-E2F1 whose disruption may contribute to cell cycle progression in gastric cancer and indicate the potential application of miR-218 in cancer therapy.