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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.

Microporous gold is common in many Te-Au deposits. Experimental formation and textures of microporous gold have recently been studied. However, few studies have worked on naturally occurring microporous gold. At the Dongping gold deposit, which is one of the most studied Au-Ag-Te deposits in China natural microporous gold is widely distributed in both primary and oxidized ores. This provides a good opportunity to study the textures and the formation of microporous gold in nature. This gold is pseudomorphous in addition to being porous, indicating that it formed via interface coupled dissolution-reprecipitation (ICDR) reactions. Four textures that were not described previously in experiments were observed in samples from this deposit. (1) A hackly interface between the microporous gold and parent calaverite. Te content shows a decreasing trend with increasing distance from the interface. This texture may result from diffusion of excess surface energy as the ICDR reaction moves towards equilibrium. (2) A less-porous band exists in the microporous gold. The band has fewer pores, smaller crystal sizes and similar chemical composition with the surrounding microporous gold. It likely formed due to the fluctuation of the fluid physicochemical conditions (including decreasing temperature, αO2(aq) and increasing pH) that changes the dissolution rate of calaverite. (3) Microporous gold precipitated between quartz and calaverite. As well, gaps developed between gold and remnant calaverite. This texture is derived from catalytic effects of a heterogeneous nucleation or recrystallization process. It is helpful for extraction and aqueous processing of Au-Te ores, and accounts for the phenomenon that quartz can accumulate gold in some hydrothermal gold deposits. (4) Petzite coexists with Ag-bearing microporous gold, indicating that petzite may also decompose to microporous gold under hydrothermal conditions. Calaverite could be dissolved without altered pyrite at high-temperature (>200 °C) conditions, but under weathering conditions, pyrite would be altered first. In primary ores, microporous gold coexists with pyrite and baryte, indicating that it was formed during the late-stage mineralization. In oxidized ores, microporous gold coexists with limonite and baryte, indicating it may have generated during late-stage mineralization and then altered by weathering. Hydrothermal fluids derive from late-stage mineralization may play the main role in the formation of microporous gold compared to weathering processes. This observation also leads us to reappraise the importance of hydrothermal fluids in the weathering dominated ICDR geological processes.

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.

Forkhead box C2 (Foxc2) protein is a transcription factor in regulation of development, metabolism, and immunology. However, the regulatory mechanisms of Foxc2 on proliferation and apoptosis of preadipocytes are unclear. In this study, we found that high-fat-diet-induced obesity elevated the expression of Foxc2 and cyclin E after 6 weeks. Additionally, Foxc2 suppressed preadipocyte differentiation, increased cell counts and augmented G1-S transition of preadipocytes, along with the elevation of cyclin E expression and the reduction levels of p27 and p53. Furthermore, Foxc2 knockdown reduced early apoptotic cells with accompanying reduction of mitochondrial membrane potential and increased fragmentation of genomic DNA. We show that Foxc2 reduces the expression of Bax, caspase-9, and caspase-3 in both serum-starved and palmitic acid-induced cell apoptotic models, which confirms the anti-apoptotic role of Foxc2. Moreover, the protein kinase B (Akt)/mammalian target of rapamycin (mTOR)C1 signaling pathway and the ERK/mTORC1 signaling pathway were activated along with preadipocyte proliferation in response to Foxc2 overexpression, whereas apoptosis marker genes were downregulated during this process. Those effects were blocked by the interference of Foxc2 or signal pathways specific inhibitors. These data collectively reveal that Foxc2 enhances proliferation of preadipocytes and inhibits apoptosis of preadipocytes by activating the Akt/mTORC1 and ERK/mTORC1 signaling pathways.

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.

The Haoyaoerhudong gold deposit in the northern margin of the North China Craton (NCC) is a large tonnage lower-grade deposit with a reserve of about 148 tons of gold. Gold mineralization is characterized by pyrite and pyrrhotite films and thin veins on the schistosity plane of the Proterozoic black shales. The orebodies, strictly controlled by a near EW-trending shear zone, are stratabound within carbonaceous phyllite and andalusite-garnet schist of the Bilute Formation. Hydrogen and oxygen isotopic data show that the ore-forming fluid was derived from a magmatic source and mixed with meteoric water. Sulfur and carbon isotope data indicate that most of the sulfur and carbon came from the black shale strata. Well-defined biotite Ar-Ar plateau age and inverse isochron age show that the deposit formed at ca. 270Ma, which suggests a probable link between Hercynian magmatism and gold mineralization. Studies on regional geology, ore geology, isotope geochemistry, and ore-forming age substantiate a complex evolutionary history of the deposit. The Proterozoic black shales rich in gold, sulfur, and organic matter, which were deposited in the Proterozoic continental margin rifts, comprised the source bed for gold mineralization. EW-ENE-structures, products of Palaeozoic orogenic process, provided pathways and mineralization space for ore-forming fluids. Hercynian tectono-magmatism and subsequent hydrothermal events remobilized gold and drove the ore-forming fluids to dilatational fracture zones. Related to postcollisional magmatic hydrothermal events, the Haoyaoerhudong gold deposit is considered as a special type of orogenic gold deposit formed in the compression–extension transition stage.

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.

// Qingda Meng 1 , Davide Valentini 2 , Martin Rao 1 , Zhenjiang Liu 1 , Shanshan Xie 1 , Ann Morgell 3 , Ernest Dodoo 1 , Matthias Lohr 3 , Elena Rangelova 3 , Marco del Chiaro 3 , Ingemar Ernberg 4 and Markus Maeurer 1, 2 1 Division of Therapeutic Immunology (TIM), Department of Laboratory Medicine (LABMED), Karolinska Institutet, Stockholm, Sweden 2 Centre for Allogeneic Stem Cell Transplantation (CAST), Karolinska University Hospital Huddinge, Stockholm, Sweden 3 Pancreatic Surgery Unit, Division of Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm, Sweden 4 Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden Correspondence to: Markus Maeurer, email: markus.maeurer@gmail.com Keywords: pancreatic cancer; mesothelin; interferon gamma; survival; antigen-specific response Received: February 27, 2017 Accepted: June 19, 2017 Published: April 27, 2018 ABSTRACT Most patients with pancreatic cancer present with extensive metastasis at diagnosis, with a 5-year survival rate of approximately 5%, despite chemotherapy and surgery. New treatment modalities are needed to improve survival. Mesothelin is a tumor-associated antigen (TAA) in patients with pancreatic cancer that could be used to gauge cellular immune responses directed against transformed cells since up to 100 percent of pancreatic ductal adenocarcinoma cells have been shown to strongly express mesothelin. A prospective, observational study was carried out in twenty-six, chemotherapy-naive patients with resectable pancreatic ductal adenocarcinoma. Participants were between 48 and 81 years (median age: 64.5 years), 15 males and 11 females. All participants were clinically followed-up between 439 and 853 days post-surgery (n=14) or until death (n=12). Peripheral blood drawn on the day of surgery was stimulated with a mesothelin peptide pool (42 peptides, non-overlapping), individual mesothelin peptides, positive (anti-CD3 antibody, OKT3) and negative controls (medium) with or without adding IL-21. Kaplan-Meier estimators were used to gauge patients’ survival pattern in relation to mesothelin-specific IFN-γ responses. A survival benefit was linked with IFN-γ responses to peptides corresponding to mature mesothelin (p=0.018) and targeted recognition of the mesothelin 601-615 epitope (MQEALSGTPCLLGPG) (p=0.006) in the presence of IL-21. Conversely, production of high levels of IFN-γ to OKT3 stimulation with IL-21 conditioning was associated with reduced survival of patients (p=0.016). Gauging anti-Mesothelin- directed immune responses will aid to identify patients i) in need of a more intensive clinical follow-up and ii) who may benefit from immunotherapeutic approaches targeting mesothelin.

Background The origin of pulmonary sinus cusp (PSC)-derived ventricular arrhythmia (VA) is a highly specialized anatomical structure; therefore, insertion of the radiofrequency ablation catheter tip to the target site to ensure safe ablation is a major challenge for clinicians. Purpose To summarize ablation methods, and provide valuable experience for the treatment of pulmonary sinus cusp-derived VA with ablation. Methods A retrospective analysis and summary of the clinical data of 15 patients undergoing catheter ablation for pulmonary sinus cusp-derived VA in Cardiac Intervention Therapy Center, The Second Xiangya Hospital of Central South University between January 2013 and July 2015 was conducted. Results For the 15 patients, the pulmonary sinus cusp-derived VA originated from the lower regions of the pulmonary sinuses, leading from the right, left and anterior sinuses of the pulmonary sinus cusp in 4, 6 and 5 patients, respectively. Nine patients with pulmonary sinus cusp-derived VAs originating from the right and anterior sinuses underwent successful reversed U-curve catheter ablation, while the other 6 cases with arrhythmias originating from the left sinus underwent successful ablation with the conventional method (non-reversed U-curve catheter ablation). All the patients were followed-up for 6–31 months, and no cases of recurrence or complications occurred. Conclusions Reversed U-curve catheter ablation is suitable for VAs originating from the right and anterior pulmonary sinus cusps, while conventional ablation can also be used for those originating from the left PSCs. This article is protected by copyright. All rights reserved

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.

Suppressor of cytokine signaling 3 (SOCS3), a signal transduction cytokine, is involved in lipid metabolism as well as in cell proliferation, differentiation, apoptosis, and so on. To explore the effects of SOCS3 on apoptosis and lipid metabolism in liver, we used a simple effective method named hydrodynamic tail vein injection to overexpress SOCS3. Then orbital blood was obtained for the assessment of blood lipid after injection. Lipid metabolism related genes were detected by Western blot after the determination of serum lipids. Meanwhile, liver cell apoptosis was observed by Hoechst and TUNEL staining and the expression of apoptosis related proteins Bax, Bcl-2, and Caspase3 were detected as well as the JAK2/STAT3 signaling pathway. In addition, we also demonstrated the effect of SOCS3 in prime hepatocyte by overexpression or interference of SOCS3 along with SD1008, which is a specific inhibitor of the JAK2/STAT3 signaling pathway. Taken together, all the results indicated that SOCS3 promoted lipid synthesis in mice liver and promoted hepatocyte apoptosis by inhibiting the activation of the JAK2/STAT3 signaling pathway, however the detailed regulation mechanism had not yet been fully understood and needs further study.

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.

The Hashitu deposit is a newly-discovered Mo deposit in the southern part of the Great Hinggan Range, NE China. Molybdenum mineralization occurs as quartz-sulfide veins within the Hashitu granite-porphyry composite pluton. The sulfide assemblage in the veins is dominated by molybdenite, with minor amounts of galena, sphalerite, chalcopyrite, pyrite and marcasite. The associated gangue minerals are quartz, fluorite, calcite, sericite, chlorite and epidote. Whole-rock chemical compositions show that the Hashitu granites belong to the A2-type. The U–Pb ages of zircons from the Hashitu granite and porphyry units are 147 ± 1 Ma and 143 ± 2 Ma, respectively. The Re–Os isochron age of molybdenites from the deposit is 150 ± 4 Ma. The molybdenite Re–Os model ages vary from 144 to 150 Ma, with a weighted mean of 147 ± 1 Ma. The results show that the ages of zircon crystallization and Mo mineralization are similar, mostly within analytical uncertainties, and that the host granite pluton is one of many late-Jurassic plutons in the Great Hinggan Range. The formation of the late-Jurassic granitic plutons in this region coincides with the subduction of the Pacific plate beneath the North China block which took place ∼2000 km to the east at the time. The occurrence of abundant late-Jurassic granitoids with compositions similar to the Hashitu pluton in the Great Hinggan Range is a positive sign for more discoveries of Mo deposits in this region.

In order to investigate the mechanism of suppressor of cytokine signaling 2 (SOCS2) on mitochondrial biogenesis, RNA interference and over-expression plasmid vectors of SOCS2 were used to transfect murine skeletal muscle C2C12 cells. Results demonstrated that over-expression of SOCS2 inhibited the differentiation of C2C12 cells, and reduced the expression of MyHC, MyoD and MyoG while elevated the protein expression of MSTN. Meanwhile the expression of PGC-1α, MDH, CPT-1 were significantly elevated in the RNA interference of SOCS2 group which were decreased in SOCS2 overexpression group. However, there was no change on the expression of UCP1 in both two groups. JC-1 dyeing showed overexpression of SOCS2 decreased the mitochondrial membrane potential and results of immunofluorescence, real-time PCR and western blotting indicated the increase expression of Cyt c, while interference SOCS2 had the opposite effects in C2C12 cells. Moreover, interference of SOCS2 elevated the p38 phosphorylation level then further increased the phosphorylation of ATF2, whereas overexpression of SOCS2 alleviated this phenomenon. Taken together, our observations indicated that SOCS2 could suppress myotube formation, act as an anti-regulator of mitochondria biogenesis via inhibiting p38 MAPK signal pathway.

We conducted a population-based case-control study in Connecticut women to test the hypothesis that genetic variations in DNA repair pathway genes may modify the relationship between body mass index (BMI) and risk of non-Hodgkin lymphoma (NHL). Compared to those with BMI

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.

Sirtuin type 1 (Sirt1) and protein kinase B (Akt2) are associated with development of obesity and inflammation, but the molecular mechanisms of Sirt1 and Akt2 interaction on adipose inflammation remain unclear. To explore these mechanisms, a mouse model was used. Mice were fed with a high-fat diet (HFD) for 8 weeks, with interventions of resveratrol (RES) or nicotinamide (NAM) during the last 15 days. The HFD reduced Sirt1 mRNA in adipose tissue and elevated interleukin-6 (IL-6) expression. RES reduced the adipose tissue weight, increased the Sirt1 mRNA level, and reduced both mRNA and protein levels of IL-6, MCP-1, inducible nitric oxide synthase, and TNF-α by inhibiting phosphorylation of Akt2 in adipose tissue. Additionally, macrophage type I marker genes were reduced while macrophage type II marker genes were elevated by RES addition. Moreover, activation of Akt2 signal by using insulin significantly blunted the inhibitory effect of RES on adipose inflammation. Immunoprecipitation assay demonstrated that RES enhances the protein-protein interaction between Sirt1 and Akt2, but NAM inhibits this interaction. Furthermore, Sirt1 significantly reduced the levels of raptor and inactivated mammalian target of rapamycin (mTOR)C1 signal by interacting with Akt2, and confirmed that RES attenuated adipose inflammation by inhibiting the mTOR/S6K1 pathway via rapamycin.

Exact periodic kink-wave solution, periodic soliton and doubly periodic solutions for the potential Kadomtsev–Petviashvii (PKP) equation are obtained using homoclinic test technique and extended homoclinic test technique, respectively. It is investigated that periodic soliton is degenerated into doubly periodic wave varying with direction of wave propagation.

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.

Rationale: This study aimed to describe the case of a 3-year-old girl with old bilateral facet dislocation on cervical vertebrae 6 and 7, who had spinal cord transection, received surgical treatment, and achieved a relative satisfactory therapeutic effect. Patient concerns: A 3-year-old girl was urgently transferred to the hospital after a car accident. Diagnoses: she was diagnosed with splenic rupture, intracranial hemorrhage, cervical dislocation, spinal transection, and Monteggia fracture of the left upper limb. Interventions: The girl underwent emergency splenectomy and was transferred to the intensive care unit of the hospital 15 days later. One-stage anterior–posterior approach surgery (anterior discectomy, posterior laminectomy, and pedicle screw fixation) was performed when the patient stabilized after 45-day symptomatic treatment. The operation was uneventful. Outcomes: The reduction of lower cervical dislocation was satisfactory, with sufficient spinal cord decompression. The internal fixation position was good, and the spinal sequence was well restored. The girl was discharged 2 weeks later after the operation and followed up for 2 years. The major nerve function of both upper limbs was recovered, with no obvious retardation of the growth of immature spine. Lessons: A satisfactory therapeutic effect was achieved for a pediatric old subaxial cervical dislocation with bilateral locked facets using anterior discectomy, posterior laminectomy, and pedicle screw fixation. The posterior pedicle screw fixation provided a good three-dimensional stability of the spine, with reduced risk and complications caused by anterior internal fixation. The growth of immature spine was not obviously affected during the 2-year follow-up.

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.

To explore the impact of different left ventricular (LV) lead pacing sites on clinical outcome of cardiac resynchronization therapy (CRT) or the device with defibrillator (CRT-D). The location of the LV lead was assessed by means of coronary venograms in right and left anterior oblique views which

New exact wave solutions including homoclinic wave, kink wave and soliton solutions for the 2D Ginzburg–Landau equation are obtained using the auxiliary function method, generalized Hirota method and the ansatz function technique under the certain constraint conditions of coefficients in equation, respectively. The result shows that there exists a kink-wave solution which tends to one and the same periodic wave solution as time tends to infinite.

In this paper, we obtain a new class of exact cross kink-wave and periodic solitary-wave solutions for Jimbo–Miwa equation by using two-soliton method, bilinear method and transforming parameters into complex ones. Moreover, we investigate singular and non-singular phenomenons of solutions. In addition, we study the resonance and non-resonance interactions between y – t periodic solitons and different line solitons.

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.

We read with great interest the paper by Di Biase et al. [(1)][1]. These investigators suggested that the recurrence of atrial fibrillation (AF) was significantly reduced by combined pulmonary vein and left atrial appendage (LAA) isolation (LAAI) for the treatment of persistent and long-standing

Melatonin is synthesized in the pineal gland and controls circadian rhythm of peripheral adipose tissue, resulting in changes in body weight. Although core regulatory components of clock rhythmicity have been defined, insight into the mechanisms of circadian rhythm-mediated proliferation in adipose tissue is still limited. Here, we showed that melatonin (20 mg/kg/d) promoted circadian and proliferation processes in white adipose tissue. The circadian amplitudes of brain and muscle aryl hydrocarbon receptor nuclear translocator-like 1 (Bmal1, P

MicroRNAs are small non-coding RNAs that partially bind to the 3’ untranslated (3’UTR) regions of target genes in animals and regulate protein production of the target transcripts. MiR-103 has been confirmed to play a critical role in lipid metabolism, however, the target genes and signaling pathway regulated by miR-103 is still unclear. In our experiment, we observed a positive function of miR-103 on the adipogenic differentiation of 3T3-L1 pre-adipocyte. Furthermore, we proved that this function of miR-103 worked through activating AKT/mTOR signal pathway and impairing target gene MEF2D. By inhibiting and over-expressing the MEF2D gene, we found that MEF2D had a negative role in regulating adipocyte key genes, and this function of MEF2D could be impaired by miR-103. In conclusion, we found that miR-103 can promote 3T3-L1 cells differentiation by targeting MEF2D and activating AKT/mTOR signal pathway. These results will shed a light on further study of microRNAs.