Search

Background Ochratoxin A (OTA) is a harmful mycotoxin and secondary metabolite produced by several species of Aspergillus and Penicillium. It is of vital importance to establish a sensitive method to detect OTA in various grains. Objective A novel, highly sensitive chemiluminescence immunoassay was developed to determine OTA in wheat, corn, and poultry feed. Methods The immunomagnetic beads were used as solid phase carriers and separators in the competitive chemiluminescence method. Anti-OTA monoclonal antibodies coated on the magnetic beads were applied as the capturing antibodies, in which OTA would compete with OTA-alkaline phosphatase in binding with OTA antibodies. Results The proposed method exhibited good linearity (R2 ≥ 0.999), adequate OTA recovery rate (83.60–102.50%), and good repeatability. The sensitivity of the proposed method was 2.05 pg/mL. In addition, the method was highly selective and showed no cross-reactions with other mycotoxins (e.g., AFB1, DON, ZEN). The developed method was applied in the analysis of wheat, corn, and two animal feeds successfully. The results obtained by the chemiluminescence method showed a high correlation with those obtained by the HPLC method (correlation coefficient 0.9958). Conclusion This strategy shows great potential for other toxic and harmful mycotoxin detection. Highlights A novel highly rapid and sensitive chemiluminescence immunoassay was established for quantitation of OTA.

In this study, a sensitive high-performance liquid chromatography method for zearalenone determination in feeds and grains was established based on the automatic immunomagnetic bead purifier analyzer (KY-AMI-I) for the one-step sample pre-treatment. Various parameters that may affect the analytical performance were investigated. Immunomagnetic bead separation system based on a monoclonal antibody (anti-ZEN antibody-immunomagnetic beads conjugation) was individually prepared for the ZEN purification. Besides, the average recoveries in spiked samples ranged from 75.23 to 102.62%, and the relative standard deviations were between 3.55 and 8.07% (

MoP is a promising anode material choice for alkali ion storage ascribed to its superior electrochemical performance. However, severe volume expansion and poor stability limited its output performance. Here, well-dispersed MoP nanoparticles are confined in carbon nanofiber skeleton by facile electrospinning method, further introduction of titania enhances the interfacial reaction and structure stability. As a result, the carbon nanofiber encapsulated MoP/TiO2 composite exhibited 150 mAh g−1 at 1 A g−1 for 4000 cycles, without obvious capacity decay. The homogenous dispersion and heterostructure construction contribute to its superior performance. This design provides a simple way for the phosphide preparation.

Background The ultrasensitive monitoring strategy of zearalenone (ZEN) is essential and desirable for food safety and human health. In the present study, a coupling of gold nanoparticles-DNA barcode and direct competitive immunoassay-based real-time polymerase chain reaction signal amplification (RT-IPCR) for ZEN close to the sensitivity of PCR-like levels is described and evaluated. Results The RT-IPCR benefited from the use of a DNA barcode and RT-PCR detection strategy, thus resulting in ultrasensitive and simple detection for ZEN. Under the optimal RT-IPCR, the linear range of detection was from 0.5 to 1000 pg mL-1 and the limit of detection was 0.5 pg mL-1 , which was 400-fold lower than the enzyme-linked immunosorbent assay. The detection procedure was simplified and the detection time was shortened. The specificity, accuracy and precision of the RT-IPCR confirmed a high performance. ZEN-positive contamination levels were from 0.056 to 152.12 ng g-1 by the RT-IPCR, which was demonstrated to be highly reliable by liquid chromatography-tandem mass spectrometry. Conclusion The proposed RT-IPCR could be used as an alternative for detecting ZEN with satisfactory ultrasensitivity, simplicity, low cost and high-throughput. The present study could provide a strategy for the ultrasensitive detection of the small molecule with a simple and practical approach, which has significant appeal and application prospects.

V-based materials are widely applied as cathode materials for aqueous zinc-ion batteries because the high potential of V5+ provides a thermodynamic basis for the transition of Zn2+. The isotropy of high entropy amorphous vanadium oxide not only avoids the performance degradation caused by the broken crystal structure, but also offers more active sites and improves the solid-state solubility and transfer kinetics of Zn2+. Herein, a high entropy amorphous vanadium oxide@carbon matrix (VxOy@C) was obtained via a simple coprecipitation method. Two electrochemical activation strategies could induce the activation of amorphous VxOy@C as a high-performance cathode material for aqueous zinc-ion batteries (AZIBs). After electrochemical activation, the amorphous VxOy@C exhibited a high specific capacity of 399.1 mA h g−1, high rate capability of 314.1 mA h g−1 at 2.0 A g−1 and excellent cycling stability for up to 300 cycles with a specific capacity retention of 91.8%. It was proven that the amorphous lump of VxOy@C transforms into an amorphous fibrous material accompanied by an increase in the valency of V. With the insertion of Zn2+, the fibrous materials would be reversibly converted into a lamellar structure.

2,5-Furandicarboxylic acid (FDCA) has the potential to replace terephthalic acid (TPA) as a building block in the preparation of bio-based and degradable polyester materials. However, the lack of efficient and stable catalysts limits the commercial viability of FDCA production. In this work, metalloporphyrins were immobilized on mesoporous SiO2 and used as a novel oxidase to convert 5-hydroxymethylfurfural (HMF) to FDCA. As the catalytic active center of P450 monooxygenases and horseradish peroxidase, supported palladium, manganese, ferri, and cobalt porphyrins via ionic-liquid-functionalized SiO2 (PdPOP-SiO2, MgPOP-SiO2, FePOP-SiO2 and CoPOP-SiO2) were able to simulate oxidase alone under mild conditions. 100% HMF (0.02 M) conversion and 94.94% FDCA yield were achieved using PdPOP-SiO2 as a biomimetic oxidase after a 12 h reaction at 373 K in water at atmospheric pressure. This type of mimetic oxidase is stable, inexpensive, and readily available, and the method developed in this work can potentially be used to oxidize HMF to FDCA in an industrial setting.

Organic porous crystals constructed by only a single kind of weak molecular interaction are invaluable to understanding the nature of the formation of organic porous materials and developing new types of porous materials. Here, we designed and synthesized two pure organic compounds of PBO and PBS through integrating planar dibenzothiophene/dibenzofuran and two phenothiazine groups together with twisted C-N bonds, which form organic microporous crystals with very good stability against strong acids and bases via pure C-H•••π interactions. Accordingly, the effective absorption of toluene has been successfully realized with adsorbing capacity of 6.20 mmol g-1, regardless of the interference of water vapour. Excitedly, these microporous materials exhibit interesting crystal-to-crystal transformation (CCT) property accompanying with the changed pore size, once being fumigated by different organic solvents. Therefore, the desorption process of toluene could be completed through simple fumigation with dichloromethane (DCM) and the second transformation of the crystal occurred in this process.

In this study, a novel P2-type sodium manganese oxide is presented using a facile preparation method. With the help of titanium doping, and introduction of interstitial water, the electrochemical p...

With the desire to realize both high capacity and high voltage, a series of materials xLi2MnO3·(1-x)NaNi1/3Co1/3Mn1/3O2 (0.5 ≥ x ≥ 0.2) are designed and prepared, which is inspired by the idea from Li-excess materials (the addition of extra lithium, manganese and charge-compensating oxygen lead to formation of structurally integrated xLi2M′O3·(1-x)LiMO2, M′ = Mn or Ti and M = Mn, Co or Ni typically). The addition of Li2MnO3 is intended to enhance the high voltage operation of sodium ternary system NaNi1/3Co1/3Mn1/3O2. X-ray diffraction patterns show that the materials are prone to form P2-type structure with the increment of Li2MnO3. Rate capability and cycle performance indicate that a more stable structure tends to be distinguishable when the content of Li2MnO3 exceeds 0.3. It is manifested that the introduction of Li2MnO3 not only plays important role in enhancing interface stability but also prevents from dramatic side reaction with electrolyte during high voltage operation.

In this work, a ratiometric fluorescent probe (RF-probe) for highly sensitive and selective detection of uric acid was reported for the first time toward H2O2 based on inner filter effect (IFE) between bimetallic gold/silver nanoclusters (Au/Ag NCs) and 2,3-diaminophenazine (DAP). For this RF-probe, uric acid was degraded to allantoin and H2O2. Upon the addition of HRP, o-phenylenediamine (OPD) could be catalytically oxidized to DAP in the presence of H2O2, then the fluorescence intensity corresponding to DAP at 580 nm increased dramatically with a fluorescence quenching of BSA-Au/Ag NCs at 690 nm, resulting in a RF-probe toward uric acid. This RF-probe allowed for the sensitive detection of uric acid in range of 5.0 × 10−6 M to 5.0 × 10−5 M with a detection limit (S/N = 3) as low as 5.1 × 10−6 M. At the same time, it has been successfully used for uric acid levels detection in human serum, and the results are consistent with those of the hospital. RF-probe built may provide a ratiometric fluorescence universal platform for detection of various species involving in the production of H2O2 in other biological systems.

A novel fluorescent sensor based on bovine serum albumin stabilized gold/silver nanoclusters (BSA-Au/Ag NCs) was developed for sensitive and facile detection of alkaline phosphatase (ALP) activity. For this fluorescent sensor, ascorbic acid 2-phosphate (AAP) was decomposed into ascorbic acid (AA) and phosphate by catalysis with ALP. The initial red fluorescence of the BSA-Au/Ag NCs was effectively quenched by KMnO4 and then the fluorescence was recovered by addition of AA. The mechanism of interaction between BSA-Au/Ag NCs and KMnO4 and AA was studied with use of the fluorescence lifetime and UV-vis absorption spectra. The results indicated that the oxidation/reduction modulated by KMnO4/AA led to surface structure destruction/restoration of the BSA-Au/Ag NCs, resulting in fluorescence quenching/recovery. The proposed fluorescence-based method based on a dark background was used to detect ALP and had excellent sensitivity, with a detection limit of 0.00076 U/L. Moreover, the method was applied to the determination of added analytes, with satisfactory recoveries (97.0–105.0 %). In a simulated eutrophic water body, this method successfully detected ALP in actual water samples and could monitor the dynamic changes of ALP activity through visual observation. More importantly, the proposed fluorescent sensor not only has the advantages of simple operation and high sensitivity but has also been successfully used on filter paper to establish a rapid and visual test paper for ALP.

Zearalenone (ZEN) has a potential hazard to human health, and is frequently found in agro-products. To minimize ZEN exposure to consumers, a novel metal-organic framework-based immunoassay system using zeolitic imidazolate framework-encapsulated horseradish peroxidase and goat anti-mouse IgG (HRP/Ab@ZIF-L) as labels was proposed for rapid and ultrasensitive detection of ZEN in agro-products. The HRP/Ab@ZIF-L not only maintained recognition ability of antibody and catalytic activity of enzyme, but also protected encapsulated proteins against high temperature, organic solvents and long term storage. Under optimal conditions, the detection limit of HRP/Ab@ZIF- L -based immunoassay reached 0.5 ng/L for ZEN, which was approximately 126-fold lower than that of conventional HRP-based immunoassay. Moreover, the proposed method showed an excellent selectivity, and a good dynamic linear detection for ZEN in the range of 0.5 ng/L to 0.476 μg/L. The recoveries of ZEN from spiked corn and wheat samples ranged from 84.50% to 96.70% with the relative standard deviation under 8.9%. In brief, the proposed immunoassay method has potential application for rapid and ultrasensitive detection of ZEN in agro-products.

Zearalenone (ZEN) is a prevalent mycotoxin that needs intensive monitoring. A semi-quantitative and quantitative immunochromatographic assay (ICA) was assembled for investigating ZEN contamination in 187 samples of cereal and their products from China in 2019. The semi-quantitative detection model had a limit of detection (LOD) of 0.50 ng/mL with visual judgment and could be completely inhibited within 5 min at 3.0 ng/mL ZEN. The quantitative detection model had a lower LOD of 0.25 ng/mL, and ZEN could be accurately and digitally detected from 0.25&ndash, 4.0 ng/mL. The ICA method had a high sensitivity, specificity, and accuracy for on-site ZEN detection. For investigation of the authentic samples, the ZEN-positive rate was 62.6%, and the ZEN-positive levels ranged from 2.7 to 867.0 ng/g, with an average ZEN-positive level being 85.0 ng/g. Of the ZEN-positive samples, 6.0% exceeded the values of the limit levels. The ZEN-positive samples were confirmed to be highly correlated using LC-MS/MS (R2 = 0.9794). This study could provide an efficiency and accuracy approach for ZEN in order to achieve visual and digitized on-site investigation. This significant information about the ZEN contamination levels might contribute to monitoring mycotoxin occurrence and for ensuring food safety.

Herein, a simple and cost-effective chemiluminescence immunoassay combined with the magnetic particles (MPCLIA) is presented for the rapid detection and analysis of Aflatoxin B1, which is a potent carcinogen found in the environment. Several physicochemical factors that potentially influence the assay performance of the proposed method were investigated in detail, including concentrations of FITC-labelled antibody, AFB1-alkaline phosphatase and magnetic particles concentration. Under the optimized experiment condition, the method showed: (i) a low limit detection of 0.05 ng/mL; (ii) satisfactory recoveries in real grain and oil samples ranging from 78% to 109%, which was well within the requirement of residue analysis, and (iii) excellent sensitivity, wide dynamic range and easy to separate in the washing process, which was superior to enzyme-linked immunosorbent assay (ELISA) method. In addition, the device used is of low cost. Moreover, the MPCLIA results for the real samples were in good agreement with that obtained by the high-performance liquid chromatography. These results suggest that the proposed MPCLIA method has potential application for screening other toxic mycotoxins occur in the food industry or other applied fields.

The potential homogeneous assay employing immunomagnetic beads (IMB) has been receiving attention as a screening tool in food-safety control; the method is simple, efficient, and does not require long incubation times or complex separation steps. In this study, a homogeneous immunoassay has been successfully developed and applied in the determination of aflatoxin B1 (AFB1) contamination in agricultural products by coupling IMB and the biotin-streptavidin (BSA) (BSA-IMB) system. Under optimal conditions, the limit of detection (LOD, IC10), half-maximal inhibition concentration (IC50) and detection range (IC20–IC80) of BSA-IMB are 0.00579, 0.573 and 0.0183–17.9 ng mL−1, respectively, for AFB1. The detection of AFB1 by BSA-IMB can be achieved in 40 min (ELISA needs at least 180 min). The cross-reactivities of BSA-IMB with its analogues are negligible (

Sodium ion batteries (SIBs) are considered as a promising candidate in the field of large-scale power grids and power storage, owing to their abundant reserves and low price. As a promising intercalated SIBs electrode, vanadium-based compounds have multiple oxidation states, rich coordination polyhedron structure and open lattice frameworks. But those materials are prone to occur the irreversible electrochemical reactions and produce metastable structural materials in the process of electrochemical reaction, resulting in a rapid decrease in capacity. Herein, a novel bilayer-structured vanadium oxide (CaV8O20·3H2O)/Carbon composite material with high specific capacity and long cycling life was synthesized. The CaV8O20·3H2O in this research has overcome the shortcomings of most vanadium-based electrode materials. In the ex-situ mechanism of this study, it was proved that the insertion of calcium ions can effectively maintain the stability of the interplanar spacing of the material and carry out sufficient redox reactions. Combined with the results of electrochemical performance comparison and mechanism analysis, a uniform carbon coating can improve the electrochemical stability and specific capacity of CaV8O20·3H2O by increasing the conductivity of the material and inhibiting adverse side effects. The reported V-based carbon composite anode material exhibited a remarkable electrochemical property with a capacity of 0.2 A g−1 and the capacity retention rate was 92.7% after 200 cycles, and the capacity retention rate was 72% after 450 cycles at a current density of 5.0 A g−1. The novel pre-intercalation model may provide a new perspective for the development of vanadium-based materials in SIBs.

Tetrabromobisphenol A bis(2-hydroxyethyl) ether (TBBPA DHEE), as one of the TBBPA derivatives, is frequently applied as a flame retardant. To date, however, little evidence has been gathered regarding its environmental behavior and fate. This is mainly attributed to the lack of an analytical approach available to measure it. Mass spectrometry cannot determine levels of TBBPA DHEE. LC-MS could not ionize TBBPA DHEE to yield enough fragments, while GC-MS could not be applied for the analysis either, due to the thermal degradation of TBBPA DHEE. We here designed a novel hapten, then conjugated it with a carrier protein and produced a type of polyclonal antibody capable of recognizing TBBPA DHEE. Based on this approach, a sensitive and high-throughput indirect competitive enzyme-linked immunosorbent assay (ELISA) was established. The respective detection limit (LOD, based on 90% B/B0) and IC50 calculated from the standard curve were 0.702 and 9.868 ng/mL for TBBPA DHEE. And the established method showed good tolerance to various matrix interferents. Using our ELISA, we measured the concentrations of these substances from environmental water samples with no pretreatment other than filtration. We found the presence of TBBPA DHEE in environmental water samples collected in Jiangsu province, and the detected concentrations ranging from 1.5 to 7.7 ng/mL. Our proposed method here could be applied to completely and systematically investigate TBBPA DHEE and TBBPA MHEE concentrations in environmental samples.

Due to the harmful nature of 4,4′-diaminobiphenyl (DABP) to the environment and human health, the determination of DABP is of great significance and has received great attention. However, there is no electrochemical method reported to detect DABP. In this paper, perylenetetracarboxylic acid (PTCA) noncovalently functionalizes carbon nanohorns (PTCA-CNHs) nanohybrids were produced using a simple method and applied for the first electrochemical sensing of DABP. Herein, PTCA has two key advantages for functionalizing CNHs in DABP analysis: (I) PTCA could improve the dispersibility of CNHs in water and (II) enhance the accumulation capacity of DABP on PTCA-CNHs surface by means of the electrostatic interactions between PTCA (negative) and DABP (positive). The results show that the oxidation current of DABP at the PTCA-CNHs modified screen printed carbon electrode (SPCE) is much higher than that of the CNHs/SPCE and bare SPCE. The PTCA-CNHs/SPCE has a linear response range of 5.0 to 400.0 nM with a detection limit of 1.8 nM towards DABP detection.

The objective of this study is to characterize the relationship between forkhead box C2 protein (Foxc2) and leptin under adipose inflammatory response.Lipopolysaccharide (LPS)-induced inflammatory model was conducted. Data from wild-type and ob/ob mice were used to compare the alternative role of leptin on Foxc2-mediated inflammation and browning. Transcriptional regulation and protein-protein interaction were analyzed by bioinformatics and proved by chromatin immunoprecipitation and co-immunoprecipitation experiment.Foxc2 and leptin correlated with inflammation and browning of white adipose tissue (WAT) in LPS-treated mice. Moreover, Foxc2-mediated inhibition of inflammation involved downstream activation of leptin signal and promoted WAT browning. We then determined CREB, the potential transcriptional factor of leptin, was required for Foxc2-mediated inflammation in the regulation of WAT browning. Foxc2 alleviated adipocyte inflammation by reducing leptin-mediated Janus-activated kinase 2/signal transducer and activator of transcription 3 (STAT3) pathway. Importantly, STAT3 physically interacted with PRDM16 and formed a complex to promote WAT browning. Exogenous Foxc2 overexpression also ameliorated inflammation and promoted adipose browning in high fat diet (HFD)-induced obese mice.Our results indicated that Foxc2 inhibited inflammation and promoted browning of WAT through positive regulation of leptin signal and the STAT3-PRDM16 complex. These findings identify a new potential means to prevent and treat obese caused metabolic syndrome of mammals.

On the basis of H2O2-mediated growth of gold nanoparticle (AuNPs), a novel plasmonic enzyme-linked immunosorbent assay (pELISA) was developed with a polyclonal antibody for the ultrasensitive simultaneous naked-eye detection of tetrabromobisphenol A bis(2-hydroxyetyl) ether (TBBPA DHEE) and tetrabromobisphenol A mono(hydroxyethyl) ether (TBBPA MHEE), one of the major derivatives and byproducts of tetrabromobisphenol A (TBBPA), respectively. In this modified indirect competitive pELISA, glucose oxidase (GOx) played an important role leading to the growth of AuNPs through a reaction between GOx and glucose to produce hydrogen peroxide (H2O2). In addition, further signal amplification was achieved via a large number of GOx molecules, which were immobilized on silica nanoparticles carrying poly brushes (SiO2@PAA) to increase the enzyme load, and the whole complex was conjugated on the second antibody. Under the optimized conditions, 10–3 μg/L TBBPA DHEE can be distinguished via the observation of a colored so...

Lanthanide ions, the excellent fluorescence markers, have the potential to simplify the performance of multi-analytes analysis. In this work, a powerful time-resolved fluoroimmunoassay (TRFIA) based on dual-labeled format were performed for detecting clenbuterol and salbutamol simultaneously in water and swine feed samples. Europium (Eu3+)-labeled goat anti-mouse IgG and Samarium (Sm3+)-labeled goat anti-rabbit IgG were prepared and used as fluorescent probes. Under optimal conditions, the half-maximal inhibition concentration (IC50) and the limit of detection (LOD, IC10) of the TRFIA were 0.263 and 0.007 ng/mL for clenbuterol, and 0.179 and 0.014 ng/mL for salbutamol, respectively. The cross-reactivities of the TRFIA between these two β2-agonists and with their analogues were negligible, thus indicated high specificity and promoted the simultaneous determination. The spiked recoveries were 72.2%–102.2% with relative standard deviations (RSDs) of 3.4–13.8% for clenbuterol and salbutamol in water and swine feed samples. Furthermore, the results of TRFIA correlated well with those obtained by HPLC. The proposed dual-labeled TRFIA was a satisfactory tool for simultaneous, sensitive, efficient and labour-saving detection of clenbuterol and salbutamol in water and swine feed samples.

The giant Shangfanggou skarn-porphyry Mo–Fe deposit, one of the largest Mo deposits in the East Qinling Orogen, is hosted by the Neoproterozoic Luanchuan Group on the southern margin of the North China Craton. The mineralization occurs mainly as veinlets and in veinlet-disseminated form within altered schists, skarns, hornfelses, and/or marbles. The hydrothermal alteration and mineralization at Shangfanggou took place over four stages. Stage 1 involved potassic alteration and silicification in the porphyry, skarnization and magnetization in carbonate strata, and hornfelsization in schists. Quartz–molybdenite veins associated with phyllic alteration were produced in the porphyry during stage 2, along with sericitization, chloritization, serpentinization, and silicification in the skarns, hornfelses and/or marbles. Quartz–polymetallic sulfide veins related to phyllic alteration were produced during stage 3, and ore-barren quartz ± calcite ± pyrite veins associated with weak carbonatation were produced during stage 4. C, O, and D isotope compositions of hydrothermal calcite, dolomite, magnetite, phlogopite, and quartz from different stages of mineralization during the formation of the Shangfanggou deposit range from − 12.9‰ to + 8.8‰, − 3.2‰ to + 22.7‰, and − 100‰ to − 80‰, respectively. Sulfur isotope compositions of sulfides from the ores of the Shanfanggou deposit range from 3.6‰ to 4.4‰. 206 Pb/ 204 Pb, 207 Pb/ 204 Pb, and 208 Pb/ 204 Pb ratios of 17.648 to 18.742 (apart from one magnetite with a value of 20.78), 15.526 to 15.690, and 38.197 to 39.823 in the Shanfanggou deposit. The evolution of the ore-forming fluids in the Shangfanggou deposit, as revealed by δ 18 O, δD, and δ 13 C isotope studies, was one of progressive modification of the magmatic fluids through fluid–wall rock interactions along conduits during fluid ascent, and this signifies the involvement of formation/strata water during the earlier stages of mineralization, and the predominance of meteoric waters over magmatic fluids in the later stages. The carbon, sulfur, and lead isotope compositions suggest mixing took place between the magmas and the host rocks within the region. Integrating the data obtained from studies of the regional geology, ore geology, fluid inclusions, and C–H–O–S–Pb isotope geochemistry, we conclude that the Shangfanggou Mo–Fe deposit is a porphyry-skarn type system that formed during a transition from compressional to extensional tectonics during the Jurassic–Early Cretaceous continental collision between the North China Craton and Yangtze Craton. The ore materials were sourced from magmatic ore-forming fluids that mixed with components derived from carbonaceous wall rocks during their ascent, and which were gradually mixed with meteoric waters, especially in the later stages.

Alpha-melanocyte stimulating hormone (αMSH) has an important role in the regulation of body weight and energy expenditure. Nevertheless, the molecular mechanisms of circulating αMSH on preadipocyte proliferation remain elusive. We found αMSH was reduced by high fat diet (HFD) while leptin was elevated in adipose tissue. Leptin resistance and endoplasmic reticulum (ER) stress of adipose tissue were increased in obese mice. αMSH increased leptin sensitivity and alleviated ER stress along with increased p-STAT3 level and reduced SOCS3, GRP78, CHOP, ATF4, p27 and p53 levels. αMSH and leptin co-treatment alleviated ER stress through decreasing the levels of GRP78 and CHOP. Tunicamycin (TM) or thapsigargin (Tg) induced ER stress blunted leptin sensitivity and inhibited preadipocyte proliferation. αMSH and leptin co-treatment increased the cell number, augmented G1-S transition, elevated leptin sensitivity, and reduced ER stress; it also activated Notch1 signal and stimulated preadipocyte proliferation, whereas ER stress marker genes were decreased during this process. However, the effects of αMSH and leptin were blocked by the specific inhibitor of Notch1 signal. In summary, our data revealed αMSH enhanced leptin sensitivity and preadipocyte proliferation, meanwhile inhibited ER stress of preadipocytes by activating Notch1 signal.

A fast and ultrasensitive indirect competitive time-resolved fluoroimmunoassay (TRFIA) was developed for the analysis of paclobutrazol in environmental water and soil samples. Paclobutrazol hapten was synthesized and conjugated to bovine serum albumin (BSA) for producing polyclonal antibodies. Under optimal conditions, the 50% inhibitory concentration (IC50 value) and limit of detection (LOD, IC20 value) were 1.09μgL(-1) and 0.067μgL(-1), respectively. The LOD of TRFIA was improved 30-fold compared to the already reported ELISA. There was almost no cross-reactivity of the antibody with the other structural analogues of triazole compounds, indicating that the antibody had high specificity. The average recoveries from spiked samples were in the range from 80.2% to 104.7% with a relative standard deviation of 1.0-9.5%. The TRFIA results for the real samples were in good agreement with that obtained by high-performance liquid chromatography analyses. The results indicate that the established TRFIA has potential application for screening paclobutrazol in environmental samples.

Increased awareness of phthalic acid esters (PAEs) toxicity has given rise to a dramatic increase in concern about the determination of these contaminations in the environment. In this paper, a sensitive, selective and rapid enzyme immunoassay of ELISA based on polyclonal antibody for detecting butyl benzyl phthalate (BBP) was developed and applied in the environmental water and soil samples. The hapten of BBP was synthesized, then applied to prepare artificial antigen and produce polyclonal antibody capable of specific recognizing BBP. From the optimal standard curve of ELISA for BBP, the values of LOD (limit of detection, IC10) and IC50 were 2.5 and 79.4 ng/mL, respectively. The ELISA showed high specificity, with the cross-reactivity toward BBP analogs

In this communication, 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) funtionalized graphene nanoribbons (HP-β-CD/GNRs) were prepared for the first time by using a simple wet chemical strategy, and then, the HP-β-CD/GNRs were applied for constructing electrochemical sensors for three representative analytes (p-aminophenol, p-AP; l-tyrosine, Tyr; rhodamine B, RhB). Owing to the synergetic effects of GNRs (excellent electrochemical properties and large surface area) and HP-β-CD (high host–guest recognition and enrichment capability), the highly sensitive electrochemical sensing platforms of three analytes were established by using HP-β-CD/GNRs modified electrode, and the detection limits were 0.0008, 0.003, and 0.001 μM for p-AP, Tyr, and RhB, respectively. Keywords: Graphene nanoribbon, Cyclodextrin, Electrochemical sensor, p-Aminophenol, l-Tyrosine, Rhodamine B

An indirect competitive enzyme-linked immunosorbent assay (ELISA) for flonicamid was developed based on a polyclonal antibody. 4-Trifluoronicotinic acid (TFNA) was used as flonicamid hapten and conjugated to bovine serum albumin (BSA) to produce an immunogen and ovalbumin (OVA) to produce a coating antigen. Polyclonal antibodies against flonicamid were successfully produced by immunizing New Zealand white rabbits. Under the optimal conditions, the 50% inhibitory concentration (IC50 value) of standard curves was 7.89 mg L−1 and the limit of detection (IC10) was 0.063 mg L−1. There was almost no cross-reactivity of the antibody with three structurally related metabolites, indicating that the antibody had high specificity. The recoveries from spiked water, soil, tomato and apples were in the range of 82.4–113.4% with relative standard deviations of 2.9–10.3%. Moreover, the ELISA for spiked samples showed reliability and high correlation with gas chromatography. These results suggest that the proposed ELISA method has potential application for screening flonicamid in agricultural and environmental samples.

Cytomegalovirus (CMV) infection remains a life-threatening condition in individuals with a suppressed immune system. CMV may also represent a clinically relevant target for immune responses in CMV-positive malignancies. We established a protocol to expand CMV-specific T cells (CMV-T) using peripheral blood mononuclear cells (PBMCs). PBMCs from 16 HLA-A*0201 donors were cultured with a cytokine cocktail comprising IL-2/IL-15/IL-21 along with overlapping peptides from CMV-pp65. Ten days later, T cells were stimulated with anti-CD3 (OKT3) and irradiated autologous PBMCs. CMV-T were detected by HLA-A*0201 CMV-pp65NLVPMVATV wild type and q226a mutant tetramers (for high-affinity T cells), intracellular cytokine staining, a CD107a mobilization assays as well as IFN-γ and TNF-α production in cell culture supernatants. We reliably obtained 50.25 ± 27.27% of CD8+ and 22.08 ± 21.83% of CD4+ T cells post-CMV-pp65 stimulation of PBMCs with a Th1-polarized phenotype and decreased Th2/Th17 responses. Most CD3 + CD8 + tetramer+ T cells were effector-memory cells, particularly among high-affinity CMV-T (q226a CMV-tetramer+). High-affinity CMV-T cells, compared to WT-tetramer+ cells, expressed higher IL-21R and lower FasL post-stimulation with CMV-pp65. The IL-2/IL-15/IL-21 cocktail also promoted CCR6 and CXCR3 expression necessary for T-cell migration into tissues. We have optimized methods for generating high-affinity CMV-specific T cells that can be used for adoptive cellular therapy in clinical practice.

Owing to the harmful nature of 4-nitrochlorobenzene (4-NCB, one toxic organic pollutant) and the low cost, high sensitivity and ease of operation of electrochemical method, it is highly desirable to develop effect electrochemical sensor for the detection of 4-NCB. Herein, by partially unzipping carbon nanohorns (CNHs) via a simple wet-chemistry method, CNHs and graphene oxide (GO) (CNHs/GO) nanohybrids were produced for sensing 4-NCB with synergistic properties. While the retained CNHs offer a path for rapid electron transport, the GO sheets formed by partially unzipping CNHs provide abundant active sites, further increase in surface area as well as improved dispersibility of the CNHs/GO. Our results show that the CNHs/GO modified electrode has excellent sensitivity to 4-NCB with a wide linear response range and a detection limit as low as 10 nM.

Leptin is an adipocyte-derived hormone and maintains adipose function under challenged conditions. Autophagy is also essential to maintain cellular homeostasis and regulate characteristics of adipose tissue. However, the effects of leptin on autophagy of adipocyte remain elusive. Here, we demonstrated endoplasmic reticulum (ER) stress and leptin were correlated with autophagy and inflammation by transcriptome sequencing of adipose tissue. Leptin-mediated inhibition of autophagy was involved in upstream reduction of ER stress proteins such as Chop, GRP78, and Atf4, since blockage of autophagy using pharmacological approach had no effect on tunicamycin-induced ER stress. Moreover, we determined KLF4, the potential transcriptional factor of Atf4, was required for the leptin-mediated autophagy in the regulation of adipocyte inflammation. Importantly, ATF4 physically interacted with ATG5 and subsequently formed a complex to promote adipocyte autophagy. Further analysis revealed that Atg5, a core component of autophagosome, was the target for leptin-mediate autophagy. In addition, leptin alleviated ER stress-induced inflammation by reducing autophagy-mediated degradation of IκB in adipocytes. Exogenous leptin treatment also ameliorated autophagy and inflammation of white adipose tissue in ob/ob mice. Taken together, our results indicated that leptin inhibited ER stress-mediated autophagy and inflammation through the negatively regulation of Atf4/Atg5 complex in adipocytes. These findings identify a new potential means for intervention of autophagy to prevent or treat obese caused metabolic syndrome of mammals.

The suppressor of cytokine signaling 3 (SOCS3) is an established negative feedback regulation transcription factor associated with leptin, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and growth hormone (GH). However, the regulatory mechanism of SOCS3 on inflammation and apoptosis of adipocyte is still not clear. In this study, we found an increased expression of adipocyte inflammatory cytokines TNF-α and IL-6 due to leptin treatment. Meanwhile Caspase3, a key executioner of apoptosis, was also elevated in this process. In addition, we observed that SOCS3 could promote inflammation whereas SOCS3 interference reversed this effect in LPS-induced adipocytes inflammatory model. Moreover, the expression of apoptosis-associated genes Bax, cleaved-Caspase9 and cleaved-Caspase3 were elevated along with decreased Bcl-2 expression as detected with Western blot and ELISA assay. The phosphorylation level of JAK2/STAT3 signal was inhibited by SOCS3 along with the elevated expression of IL-6, TNF-α and Caspase3. We also demonstrated that stable knockdown of SOCS3 along with SD1008, a specific inhibitor of JAK2/STAT3 signaling pathway, could significantly inhibit inflammation and apoptosis of adipocyte. Altogether, these results inferred that SOCS3 promotes adipocyte apoptosis by both aggravating inflammation and inhibiting the activity of JAK2/STAT3 signaling pathway.

Multiwalled carbon nanotubes (MWCNT) were covered with reduced graphene oxide nanoribbons (rGONR) to give a material with a core-shell heterostructure of the type MWCNT@rGONR. It was obtained by (a) longitudinal partial unzipping of MWCNT to form MWCNT@GONR, and (b) subsequent chemical reduction with hydrazine to give MWCNT@rGONR. The MWCNT@rGONR heterostructure was used to modify a glassy carbon electrode (GCE) to obtain an electrochemical sensor for p-dihydroxybenzene (DHB). The synergistic effects of the MWCNT and the rGONR results in a distinctly improved redox current towards DHB compared to a bare GCE, an MWCNT/GCE, and an MWCNT@GONR/GCE. At the working voltage range from −100 to 400 mV, it displays a linear response to DHB in the 80 to 3000 nM concentration range with a 20 nM detection limit.

Salbutamol (SAL) has potential hazardous effects on health, and has been found in natural water globally. To protect consumers from risks related to SAL residues, a fast and highly sensitive enzyme-linked immunosorbent assay (ELISA) was developed for the detection of SAL in environmental water samples. SAL hapten was synthesized and conjugated to proteins to be used as the immunogen for producing polyclonal antibodies. Under optimized conditions, the ELISA showed a 50% inhibitory concentration (IC50 value) of 0.466μg/L, with a limit of detection (LOD, IC10 value) of 0.021μg/L. The LOD of the ELISA was 5-fold lower than the maximum residue limits. The average recoveries of SAL from environmental samples, including tap water, river water and wastewater, ranged from 80.1 to 115.6%, which is well within the requirements of residue detection. Moreover, the results of the ELISA correlated well with high-performance liquid chromatography results, with a correlation coefficient of 0.988. These results suggest that the proposed ELISA method has potential application for the monitoring of SAL residues in environmental water samples.

Background Information Microtubule affinity-regulating kinase 4 (MARK4) deficiency has been reported to negatively regulate diet-induced obesity and to mitigate insulin resistance in knockout mice, and thus may play a role in metabolic syndrome. However, the details of the molecular mechanism have yet to be revealed and the impacts of MARK4 on apoptosis remain unexplored. This study investigated the role of Mark4 in the regulation of lipid accumulation and apoptosis in adipocytes and analysed signalling pathways involved. Results We found that Mark4 significantly up-regulated the expression of gene sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS), acetyl-CoA carboxylase-α (ACCα) and peroxisome proliferator activated receptor-γ (PPARγ); and reduced the protein contents of adipose triglyceride lipase (ATGL), as evidenced by the dramatic increasing lipid droplet accumulation in 3T3-L1 cells. Furthermore, a terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) apoptosis assay showed that Mark4 triggered apoptosis of adipocytes; and apoptosis was confirmed by the decreased protein contents of B-cell lymphoma-2 (Bcl-2), full-length caspase-3 and full-length caspase-9, as well as the increased expression of Bax, cleaved caspase-3 and cleaved caspase-9. Analysis of special inhibitors allowed us to offer the following explanation for these impacts of Mark4: activation of Jun N-terminal kinase1 (JNK1) promoted both apoptosis and adipogenesis, whereas inhibition of the p38 mitogen-activated protein kinase (p38MAPK) pathway contributed to lipid accumulation alone. Conclusions Mark4 promotes adipogenesis in 3T3-L1 adipocytes by activating the JNK1 and inhibiting the p38MAPK pathway, and triggers apoptosis by activating the JNK1 pathway. We conclude that anti-Mark4 therapy targetted to inhibit lipid accumulation and apoptosis of adipocytes shows potential as a novel therapeutic strategy for treatment of obesity-associated metabolic complications.

Background The modification of cardiac energy metabolism during atrial fibrillation (AF) has been demonstrated in previous studies, indicating a close association between these 2 processes. The aim of the present study was to identify the underlying mechanisms via profiling of the expression of energy metabolism-related proteins in the left atrial appendage (LAA) of patients with AF. Methods and results Isobaric tag for relative and absolute quantification-coupled 2-D liquid chromatography-tandem mass spectrometry (iTRAQ-coupled 2-D LC-MS/MS) was used to profile the expression of energy metabolism-related proteins in the LAA from valvular disease patients with sinus rhythm (SR; n=6) and AF (n=8). Using ProteinPilot 4.0, 122 energy metabolism-related proteins, consisting of 39 carbohydrate metabolism-related proteins, 22 proteins involved in lipid metabolism, 49 biological oxidation-related proteins and 12 other kinds of proteins, were identified. Most of them were key enzymes involved in energy metabolism. Moreover, most of the proteins that were expressed differently in the LAA between the AF and SR patients, and which were related to energy metabolism, were downregulated. These results were further validated on western blot. Conclusions Atrial myocardium energy production in valvular disease patients is impaired during permanent AF, and this impairment in energy production may be involved in the matrix of AF formation.

Alpha melanocyte stimulating hormone (α-MSH) and Forkhead box C2 protein (Foxc2) enhance lipolysis in multiple tissues. However, their relationship in adipose fatty acid oxidation (FAO) remains unclear. Here, we demonstrated that α-MSH and Foxc2 increased palmitate oxidation to CO2 in white (WAT) and brown adipose tissue (BAT). C/EBPβ expression was reduced by α-MSH and Foxc2. FFA level was elevated by α-MSH and pc-Foxc2 treatment along with increased FAO in white and brown adipocytes. The expression of FAO key enzymes, medium-chain acyl-CoA dehydrogenase (MCAD) and long-chain acyl-CoA dehydrogenase (LCAD) were increased in α-MSH and pc-Foxc2 group. Combination of α-MSH and Foxc2 treatment synergistically promoted FAO through increasing the activity of CPT-1 and phosphorylation of ACC. We found C/EBPβ bind to MC5R and Foxc2 promoter regions and inhibited FAO. cAMP level was increased by α-MSH and Foxc2 individually treated or combined treatment. Furthermore, cAMP/PKA pathway-specific inhibitor (H89) blocked the FAO, despite in α-MSH and Foxc2 both added group. While forskolin, the cAMP agonist, promoted FAO and enhanced the effect of α-MSH and Foxc2. Collectively, α-MSH and Foxc2 mutual promote FAO in WAT and BAT via cAMP/PKA signal pathway. And C/EBPβ as a transcription suppressor inhibits α-MSH and Foxc2 expression and FAO.

A simple and signal-amplified, label-free electrochemical impedimetric immunosensor for dibutyl phthalate (DBP), a type of phthalate ester, was developed using gold nanoparticles (AuNPs) induced signal amplification. The approach was based on an indirectly competitive binding system of coating antigen that coated on antigen/chitosan/MWCNTs@GONRs/GCE (modified electrodes), target (DBP) and anti-DBP antibody. Signal amplification was performed via the enlargement of AuNPs through NADH-promoted catalytic precipitation, which caused a marked increase in the electron-transfer resistance and electrostatic repulsion after an AuNP-labeled second antibody conjugated to anti-DBP. Under the optimized conditions, this method displayed: (i) low detection limits (7 ng/mL, 10 times lower than the traditional ELISA method using the same antibody); (ii) satisfactory accuracy (recoveries, 86.0–120.4%) and agreement with corresponding ELISA method; (iii) high tolerance to some environmental interferents; and (iv) low cost and low sample consumption (6 µL). Our results demonstrate the great potential and high efficiency of this immunosensor in practical applications for the cost-effective and sensitive monitoring of DBP in aquatic environments.

MAP/Microtubule affinity-regulating kinase 4 (Mark4) plays an important role in the regulation of microtubule organization, adipogenesis and apoptosis. However, the role of Mark4 plays in oxidative stress and inflammation are poorly understood. In this study, we found Mark4 was induced by high fat diet (HFD) while PPARγ was elevated significantly in mice adipocytes. Further analyses revealed Mark4 impaired mitochondrial oxidative respiration and increased reactive oxygen species (ROS) production. At same time, the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) were greatly reduced. By treating cells with H2O2 and vitamin E (VE), Mark4 accentuated oxidative stress along with increased mRNA level of inflammatory factor interleukin-6 (IL-6) and decreased leptin mRNA. Furthermore, we found PPARγ bind to Mark4 promoter region and inhibited Mark4 expression. We showed PPARγ interacted with Mark4 and inhibited the stimulating effect of Mark4 on oxidative stress and inflammation. Finally, we demonstrated that the IKKα/NF-κB signal pathway was involved in Mark4 induced oxidative stress and inflammation, while PTDC, a special inhibitor of NF-κB signal pathway, reduced oxidative stress and inflammation. Thus, our study indicated that Mark4 was a potential drug target for treating metabolic diseases.

Generation of T lymphocytes with reactivity against cancer is a prerequisite for effective adoptive cellular therapies. We established a protocol for tumor-infiltrating lymphocytes (TILs) from patients with pancreatic ductal adenocarcinoma. Tumor samples from 17 pancreatic cancer specimens were cultured with cytokines (IL-2, IL-15, and IL-21) to expand TILs. After 10 days of culture, TILs were stimulated with an anti-CD3 antibody (OKT3) and irradiated allogeneic peripheral blood mononuclear cells. Reactivity of TILs against tumor-associated antigens (mesothelin, survivin, or NY-ESO-1) was detected by intracellular cytokine production by flow cytometry. Cytotoxicity was measured using a Chromium 51 release assay, and reactivity of TILs against autologous tumor cells was detected by INF-[gamma] production (ELISA). TIL composition was tested by CD45RA, CCR7, 4-1BB, LAG-3, PD-1, TIM3, and CTLA-4 marker analysis. TCR V[beta] was determined by flow cytometry and TCR clonality was gauged measuring the CDR3 region length by PCR analysis and subsequent sequencing. We could reliably obtain TILs from 17/17 patients with a majority of CD8(+) T cells. CD3(+)CD8(+), CD3(+)CD4(+), and CD3(+)CD4(-)CD8(-)[double-negative (DN) T cells] resided predominantly in central (CD45RA(-)CCR7(+)) and effector (CD45RA-CCR7-) memory subsets. CD8(+) TILs tested uniformly positive for LAG-3 (about 100%), whereas CD4(+) TILs showed only up to 12% LAG-3(+) staining and PD-1 showed a broad expression pattern in TILs from different patients. TILs from individual patients recognized strongly (up to 11.9% and 8.2% in CD8(+)) NY-ESO-1, determined by ICS, or mesothelin, determined respectively by TNF-[alpha] and IFN-[gamma] production. Twelve of 17 of CD8(+) TILs showed preferential expansion of certain TCR V[beta] families (eg, 99.2% V[beta]13.2 in CD8(+) TILs, 77% in the V[beta]1, 65.9% in the V[beta]22, and 63.3% in the V[beta]14 family). TCR CDR3 analysis exhibited monoclonal or oligoclonal TCRs, some of them (eg, CD8(+) V[beta]13.2) reacting strongly against autologous tumor defined by INF-[gamma] production or by cytotoxicity. We have optimized methods for generating pancreatic cancer–specific TILs that can be used for adoptive cellular therapy of patients with pancreatic cancer.

An enzyme-linked immunosorbent assay (ELISA) was developed based on a polyclonal antibody for the analysis of thiacloprid in agricultural samples. Thiacloprid hapten was synthesized and conjugated to bovine serum albumin to produce an immunogen and ovalbumin to produce a coating antigen. Polyclonal antibodies were obtained from immunized New Zealand white rabbits. Under optimal conditions (5 % methanol, 0.1 mol/L Na+, pH 5.5), the ELISA showed a 50 % inhibitory concentration (IC50) value of 0.01 mg/L and a limit of detection (IC10) of 0.47 μg/L. No obvious cross-reaction with the other structural analogs of neonicotinoid insecticides showed that the polyclonal antibodies had a high specificity for thiacloprid. The average recoveries from spiked water, soil, pear, and tomato were in the range of 80 to 119 %. The results of the ELISA were confirmed by high-performance liquid chromatography, and the correlation of the results from the two methods had a high correlation coefficient of 0.99 (n = 3) in spiked samples (soil, pear, and tomato). The proposed ELISA could successfully be applied to the determination of thiacloprid residues in agricultural samples.

The Jiawula Ag–Pb–Zn deposit lies in the renowned Ag–polymetallic metallogenic province of northern China. The origin of this structurally controlled ore body is linked to fluid migration and mineralization along cogenetic fault systems. Sulphur isotopic compositions suggest that the ore-forming aqueous solutions were derived mainly from deep magmatic fluids. Hydrogen and oxygen isotopic compositions indicate that these fluids were magmatic during early stages of ore formation and meteoric during late-stage mineralization. Lead isotopic compositions indicate that this metal was derived mainly from a mantle source, and to a lesser extent from a crustal source. Collectively, the isotopic data indicate that formation of the Jiawula Ag–Pb–Zn deposit was ultimately a reflection of late Yanshanian (140–120 Ma) volcanic–subvolcanic hydrothermal activity. The addition of meteoric water to these magmatic hydrothermal fluids created favourable conditions for mineralization. During ore formation, metallogenesis took...

Cardiovascular disease associated with oxidative stress, including atherosclerosis, is the leading cause of mortality worldwide. The accelerated proliferation and migration of vascular smooth muscle cells are the predominant characteristics of atherogenesis, and endothelial dysfunction is a major risk factor for the pathogenesis of atherosclerosis. Podocalyxin (PODXL), a type I member of the cluster of differentiation 34 family of sialomucins, functions as a pro-adhesive molecule. Emerging evidence has revealed the importance of micro (mi)RNAs in the cardiovascular system. The present study demonstrated that there was an inverse association between miRNA (miR)-125b and PODXL in human umbilical vein endothelial cells and human aortic vascular smooth muscle cells (HAVSMCs) treated with oxidized low‑density lipoprotein (LDL) and platelet derived growth factor. Additionally, miR-125b had a suppressive function in cell proliferation and migration, at least partially via targeting PODXL in the HAVSMCs. Furthermore, the data suggested that the functions of miR-125b in arteriosclerosis obliterans may be associated with transgelin, lectin-type oxidized LDL receptor-1, vascular endothelial-cadherin, intercellular adhesion molecule-1, interleukin-6 and monocyte chemotactic protein-1. In conclusion, miR-125b was found to be important in arteriosclerosis obliterans by suppressing the expression of PODXL and may serve as a potential therapeutic target for the treatment of arteriosclerosis obliterans.

Hanjiangite, ideal formula Ba 2 Ca(V 3+ Al)[Si 3 AlO 10 (OH) 2 ]F(CO 3 ) 2 , is a new mineral found in the Shiti barium deposit in the Dabashan region, China. Hanjiangite was collected from lenticular witherite ore bodies and witherite-barite-quartz veins cutting carbonaceous slates. It is found as disseminations in both settings. It occurs as thin, euhedral, tetragonal, tabular crystals, and anhedral grains. Grain size commonly varies from 0.05 to 4 mm and occasionally reaches 15 mm. It is yellow-green or dark green, with one well-developed or perfect cleavage on {001}. Other physical properties are: brittle, transparent to translucent, nonfluorescent, vitreous luster, greenish white streak, hardness 4 in the Mohs scale, splintery fracture, and a measured density of 3.69 g/cm 3 . Hanjiangite is biaxial (−), α = 1.615, β = 1.655, γ = 1.700 (589 nm), and displays strong pleochroism from pale yellow-green to dark green. It is monoclinic, with unit-cell parameters refined from single-crystal X-ray diffraction: a = 5.2050(12), b = 9.033(2), c = 32.077(8) A, β = 93.49(8)°, V = 1505.4(8) A 3 , Z = 4, space group C 2. The strongest seven lines of the X-ray powder-diffraction pattern [ d in A( I )( hkl )] are: 15.866(7)(002), 5.340(91)(006), 4.010(10)(1 14), 3.209(23) (027), 2.676(100) (1110), 2.294(29)(137), and 2.008(11)(228). Chemical analysis by a combination of electron microprobe, HF combustion, IR absorption, thermogravimetric analysis (TGA) and crystal-structure refinement gives SiO 2 19.64, TiO 2 1.12, Al 2 O 3 11.19, MgO 0.54, CaO 4.91, BaO 34.89, V 2 O 3 9.93, Cr 2 O 3 1.75, Na 2 O 0.20, K 2 O 0.06, MnO 0.01, FeO 0.02, NiO 0.01, SrO 0.34, Y 2 O 3 0.85, La 2 O 3 0.14, Nd 2 O 3 0.26, F 1.80, Cl 0.04, CO 2 10.37, H 2 O 1.30, F=O –0.76, Cl=O –0.01, sum 98.60 wt%. The empirical formula is (Ba 1.98 Na 0.06 K 0.01 ) ∑2.05 (Ca 0.76 Mg 0.12 Y 0.06 Sr 0.03 La 0.01 Nd 0.01 ) ∑0.99 (V 1.15 Al 0.75 Cr 0.20 Ti 0.12 ) ∑2.22 [(Si 2.84 Al 1.16 ) ∑4.00 O 10 ][(OH) 1.25 O 0.77 ] ∑2.02 (F 0.82 Cl 0.01 ) ∑0.83 (CO 3 ) 2.05 based on 19 anions per formula unit. A single-crystal X-ray structure determination (sample 2005st-17) was carried out with a Smart APEX CCD system using Mo K α (λ = 0.71073 A) radiation. The structure was solved using direct methods, and refined with the SHELXTL PC (Bruker AXS Inc.) package. Anisotropic refinement using all measured independent data and reflections with I > 2σ( I ) resulted in an R 1 factor of 0.08 and wR 2 of 0.20. The crystal structure is composed of alternating T-O-T and Ba 2 Ca(CO 3 ) 2 F layers. The Ba 2 Ca(CO 3 ) 2 F layer occurs between the T-O-T layers. The T site contains both Si and Al, and the O site both V 3+ and Al. The interlayer hosts not only Ba (like in chernykhite) but also Ca atoms and (CO 3 ) groups. While the coordination of the cations in the TOT layer is obvious, the coordinations of Ca and Ba in the interlayer are not straight forward. A hanjiangite has three polytypes, namely 1 M -type, 2 M -type, and 3 T -type.

A fast and sensitive polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) was developed for the analysis of flonicamid in environmental and agricultural samples. Two haptens of flonicamid differing in spacer arm length were synthesized and conjugated to proteins to be used as immunogens for the production of polyclonal antibodies. To obtain most sensitive combination of antibody/coating antigen, two antibodies were separately screened by homologous and heterologous assays. After optimization, the flonicamid ELISA showed that the 50% inhibitory concentration (IC50 value) was 3.86mgL(-1), and the limit of detection (IC20 value) was 0.032mgL(-1). There was no cross-reactivity to similar tested compounds. The recoveries obtained after the addition of standard flonicamid to the samples, including water, soil, carrot, apple and tomato, ranged from 79.3% to 116.4%. Moreover, the results of the ELISA for the spiked samples were largely consistent with the gas chromatography (R(2)=0.9891). The data showed that the proposed ELISA is an alternative tool for rapid, sensitive and accurate monitoring of flonicamid in environmental and agricultural samples.

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for the herbicide haloxyfop-P-methyl is reported. Two haptens of haloxyfop-P-methyl with different spacer-arm lengths were synthesized, to which polyclonal antibodies were obtained by immunizing New Zealand rabbits. The most sensitive combination of antibody/coating antigen was selected by homologous and heterologous assays. The results indicated that the spacer-arm length of hapten can affect assay sensitivity. The optimized ic-ELISA showed that the 50% inhibitory concentration (IC(50)) was 0.0419 mg/L, and the limit of detection (LOD) was 0.0019 mg/L. Recoveries were obtained from the agricultural samples, including water, soil, cucumbers, cabbages, tomatoes, and potatoes; all were in the range of 87.4%-111.8%, well within the requirements of residue detection.

Obesity is associated with macrophage infiltration and metabolic inflammation, both of which promote metabolic disease progression. Melatonin is reported to possess anti-inflammatory properties by inhibiting inflammatory response of adipocytes and macrophages activation. However, the effects of melatonin on the communication between adipocytes and macrophages during adipose inflammation remain elusive. Here, we demonstrated melatonin alleviated inflammation and elevated α-ketoglutarate (αKG) level in adipose tissue of obese mice. Mitochondrial isocitrate dehydrogenase 2 (Idh2) mRNA level was also elevated by melatonin in adipocytes leading to increase αKG level. Further analysis revealed αKG was the target for melatonin inhibition of adipose inflammation. Moreover, sirtuin 1 (Sirt1) physically interacted with IDH2 and formed a complex to increase the circadian amplitude of Idh2 and αKG content in melatonin-inhibited adipose inflammation. Notably, melatonin promoted exosomes secretion from adipocyte and increased adipose-derived exosomal αKG level. Our results also confirmed that melatonin alleviated adipocyte inflammation and increased ratio of M2 to M1 macrophages by transporting of exosomal αKG to macrophages and promoting TET-mediated DNA demethylation. Furthermore, exosomal αKG attenuated signal transducers and activators of transduction-3 (STAT3)/NF-κB signal by its receptor oxoglutarate receptor 1 (OXGR1) in adipocytes. Melatonin also attenuated adipose inflammation and deceased macrophage number in chronic jet-lag mice. In summary, our results demonstrate melatonin alleviates metabolic inflammation by increasing cellular and exosomal αKG level in adipose tissue. Our data reveal a novel function of melatonin on adipocytes and macrophages communication, suggesting a new potential therapy for melatonin to prevent and treat obesity caused systemic inflammatory disease.

Fatty acid binding protein 4 (FABP4), plays key role in fatty acid transportation and oxidation and increases with leptin synergistically during adipose inflammation process. However, the regulation mechanism between FABP4 and leptin on mitochondrial fatty acid oxidation remains unclear. In this study, we found that FABP4 reduced the expression of leptin, CPT-1 and AOX1 in mice adipocytes. Conversely, FABP4 was down-regulated in a time-dependent manner by leptin treatment. Additionally, forced expression of FABP4 attenuated the expression of PGC1-α, UCP2, CPT-1, AOX1 and COX2 compared with leptin incubation. Moreover, mitochondrial membrane potential, fatty acid oxidation enzyme medium-chain acyl-CoA dehydrogenase (MCAD), long-chain acyl-CoA dehydrogenase (LCAD) and Cyt C levels were reduced in response to the overexpression of FABP4. These reductions correspond well with the reduced release of free fatty acid and the inactivation of mitochondrial complexes I and III by FABP4 overexpression. Furthermore, addition of the Akt/mTOR pathway-specific inhibitor (MK2206) blocked the mitochondrial fatty acid oxidation and respiration factors, whereas interference of FABP4 overcame these effects. Taken together, FABP4 could reverse the activation of the leptin-induced mitochondrial fatty acid oxidation and the inhibition of Akt/mTOR signal pathway played a key role in this process.

Pyroptosis is a proinflammatory form of cell death that is associated with pathogenesis of many chronic inflammatory diseases. Melatonin is substantially reported to possess anti-inflammatory properties by inhibiting inflammasome activation. However, the effects of melatonin on inflammasome-induced pyroptosis in adipocytes remain elusive. Here, we demonstrated that melatonin alleviated lipopolysaccharides (LPS)-induced inflammation and NLRP3 inflammasome formation in mice adipose tissue. The NLRP3 inflammasome-mediated pyroptosis was also inhibited by melatonin in adipocytes. Further analysis revealed that gasdermin D (GSDMD), the key executioner of pyroptosis, was the target for melatonin inhibition of adipocyte pyroptosis. Importantly, we determined that nuclear factor κB (NF-κB) signal was required for the GSDMD-mediated pyroptosis in adipocytes. We also confirmed that melatonin alleviated adipocyte pyroptosis by transcriptional suppression of GSDMD. Moreover, GSDMD physically interacted with interferon regulatory factor 7 (IRF7) and subsequently formed a complex to promote adipocyte pyroptosis. Melatonin also attenuated NLRP3 inflammasome activation and pyroptosis, which was induced by LPS or obesity. In summary, our results demonstrate that melatonin alleviates inflammasome-induced pyroptosis by blocking NF-κB/GSDMD signal in mice adipose tissue. Our data reveal a novel function of melatonin on adipocyte pyroptosis, suggesting a new potential therapy for melatonin to prevent and treat obesity caused systemic inflammatory response.

Adiponectin is a cytokine produced predominantly by adipose tissue and correlates with glucose and lipid homeostasis. However, the effects of adiponectin on endoplasmic reticulum (ER) stress and apoptosis of adipose tissue remain elusive. In this study, we found that tunicamycin-induced ER stress increased serum free fatty acid (FFA) and impaired glucose tolerance, elevated the mRNA levels of GRP78, Chop, ATF2 and caspase 3, but reduced adiponectin mRNA level in white adipose tissue. Moreover, ER stress-triggered adipocyte apoptosis by increasing cellular FFA level and Ca2+ level. Further analysis revealed that adiponectin alleviated ER stress-induced adipocyte apoptosis by elevating peroxisome proliferator-activated receptor alpha (PPARα) mRNA level. Our data also confirmed that adiponectin reduced early apoptotic cells and blocked the mitochondrial apoptosis pathway by activating the AdipoR1/AMP-activated protein kinase (AMPK) signal pathway. In addition, PPARα bound to ATF2 promoter region and inhibited transcription of ATF2. The inhibition of adipocyte apoptosis by adiponectin was correlated with transcriptional suppression of ATF2. Furthermore, adiponectin inhibited ER stress-induced apoptosis by activating the AMPK/PKC pathway. In summary, our data demonstrate adiponectin inhibited ER stress and apoptosis of adipocyte in vivo and in vitro by activating the AMPK/PPARα/ATF2 pathway. Our study establishes that adiponectin is an important adipocytokine for preventing and treating obesity.