Search

The G protein–coupled bile acid receptor (GPBAR) is the membrane receptor for bile acids and a driving force of the liver–bile acid–microbiota–organ axis to regulate metabolism and other pathophysiological processes. Although GPBAR is an important therapeutic target for a spectrum of metabolic and neurodegenerative diseases, its activation has also been found to be linked to carcinogenesis, leading to potential side effects. Here, via functional screening, we found that two specific GPBAR agonists, R399 and INT-777, demonstrated strikingly different regulatory effects on the growth and apoptosis of non–small cell lung cancer (NSCLC) cells both in vitro and in vivo. Further mechanistic investigation showed that R399-induced GPBAR activation displayed an obvious bias for β-arrestin 1 signaling, thus promoting YAP signaling activation to stimulate cell proliferation. Conversely, INT-777 preferentially activated GPBAR-Gs signaling, thus inactivating YAP to inhibit cell proliferation and induce apoptosis. Phosphorylation of GPBAR by GRK2 at S310/S321/S323/S324 sites contributed to R399-induced GPBAR–β-arrestin 1 association. The cryoelectron microscopy (cryo-EM) structure of the R399-bound GPBAR-Gs complex enabled us to identify key interaction residues and pivotal conformational changes in GPBAR responsible for the arrestin signaling bias and cancer cell proliferation. In summary, we demonstrate that different agonists can regulate distinct functions of cell growth and apoptosis through biased GPBAR signaling and control of YAP activity in a NSCLC cell model. The delineated mechanism and structural basis may facilitate the rational design of GPBAR-targeting drugs with both metabolic and anticancer benefits.

Carboxysomes in cyanobacteria enclose the enzymes Rubisco and carbonic anhydrase to optimize photosynthetic carbon fixation. Understanding carboxysome assembly has implications in agricultural biotechnology. Here we analyzed the role of the scaffolding protein CcmM of the β-cyanobacterium Synechococcus elongatus PCC 7942 in sequestrating the hexadecameric Rubisco and the tetrameric carbonic anhydrase, CcaA. We find that the trimeric CcmM, consisting of γCAL oligomerization domains and linked small subunit-like (SSUL) modules, plays a central role in mediation of pre-carboxysome condensate formation through multivalent, cooperative interactions. The γCAL domains interact with the C-terminal tails of the CcaA subunits and additionally mediate a head-to-head association of CcmM trimers. Interestingly, SSUL modules, besides their known function in recruiting Rubisco, also participate in intermolecular interactions with the γCAL domains, providing further valency for network formation. Our findings reveal the mechanism by which CcmM functions as a central organizer of the pre-carboxysome multiprotein matrix, concentrating the core components Rubisco and CcaA before β-carboxysome shell formation., Biochemical, biophysical and structural analysis reveals how the scaffolding protein CcmM recruits the enzymes Rubisco and carbonic anhydrase into a condensate for encapsulation into carboxysomes—microcompartments in cyanobacteria that serve to optimize CO2 assimilation.

In this paper, in-depth analysis, and evaluation of the transient stability of a renewable energy power system are carried out, and a transient stability evaluation method based on stacked autoencoders is proposed. The method breaks through the traditional two-stage evaluation mode. The method proposes a deep network model with a multi-branch structure for different physical natures of different measured data. With the help of different sparse denoising coding networks (branches), features are extracted from each measurement separately and fused on top of the model to finally complete the transient stability assessment. The simulation results show that the method can effectively handle the simultaneous input of multiple measurements, and has the advantages of high accuracy and robustness in noisy environments. To effectively determine the model hyperparameters, the orthogonal test is used for model selection, which can significantly reduce the hyperparameter seeking space and save computational resources. The simulation results show that the method can effectively use multiple measurements to improve the accuracy of wind power prediction. Also, the input variable selection index based on mutual information can provide a good guide for further improvement of model performance.

Thienoisoindigo as a popular conjugated skeleton has been constantly applied in the construction of organic optoelectronic materials. Exploring its new applications in fluorescent sensors and bioimaging is helpful to extend its potential. In this work, a thienoisoindigo fluorophore was first selected as a building block to be applied in the construction of the near-infrared fluorescent materials with an aggregation-induced emission manner through introducing triarylamine, thiophene-bridged triarylamine, and

Using Calotropis gigantea fiber (CGF) as the bio-template, the magnetic carbonized fiber (MC-CGF) was successfully prepared by anchoring Fe3O4 onto CGF surface via pyrolysis, and well characterized by means of instrumental analysis. Subsequently, MC-CGF was used as the adsorbent to remove perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) from water. The adsorption equilibrium of PFOA and PFOS on MC-CGF could be achieved within 1 h and 2 h, with maximum adsorption capacity of ~ 200 mg/g for them at pH 3.0. Coexisting ions (Na+, Mg2+, Fe3+) promoted the adsorption efficiency of MC-CGF to PFOA/PFOS, in particular for multivalent metal cations. During the adsorption process, hydrophobic interaction was principally involved, and was considered to be the driving force for fast removal of perfluoroalkyl pollutants. Such a hydrophobic-driven interaction was further evidenced using other perfluoroalkyl pollutants with different C–F chain length and functional groups as the models. Finally, MC-CGF was easily regenerated and recycled many times, demonstrating its potential for removing perfluoroalkyl pollutants from water in the future.

BackgroundWe aimed to develop and validate a new nomogram for predicting the risk of intracranial hemorrhage (ICH) in patients with acute ischemic stroke (AIS) after intravenous thrombolysis (IVT).MethodsA retrospective study enrolled 553 patients with AIS treated with IVT. The patients were randomly divided into two cohorts: the training set (70%, n = 387) and the testing set (30%, n = 166). The factors in the predictive nomogram were filtered using multivariable logistic regression analysis. The performance of the nomogram was assessed based on the area under the receiver operating characteristic curve (AUC-ROC), calibration plots, and decision curve analysis (DCA).ResultsAfter multivariable logistic regression analysis, certain factors, such as smoking, National Institutes of Health of Stroke Scale (NIHSS) score, blood urea nitrogen-to-creatinine ratio (BUN/Cr), and neutrophil-to-lymphocyte ratio (NLR), were found to be independent predictors of ICH and were used to construct a nomogram. The AUC-ROC values of the nomogram were 0.887 (95% CI: 0.842–0.933) and 0.776 (95% CI: 0.681–0.872) in the training and testing sets, respectively. The AUC-ROC of the nomogram was higher than that of the Multicenter Stroke Survey (MSS), Glucose, Race, Age, Sex, Systolic blood Pressure, and Severity of stroke (GRASPS), and stroke prognostication using age and NIH Stroke Scale-100 positive index (SPAN-100) scores for predicting ICH in both the training and testing sets (p < 0.05). The calibration plot demonstrated good agreement in both the training and testing sets. DCA indicated that the nomogram was clinically useful.ConclusionsThe new nomogram, which included smoking, NIHSS, BUN/Cr, and NLR as variables, had the potential for predicting the risk of ICH in patients with AIS after IVT.

Series of B-doped perovskite-like materials CeCu0.5Co0.5O3 (B-C3O) were fabricated with unique ferromagnetic property due to partial substitution of non-magnetic 2p-impurities boron in the lattice. Then, B-C3O was used for activating peroxymonosulfate (PMS) for the degradation of norfloxacin (NOR), one kind of emerging pollutants with the concentration level up to mg/L in wastewaters. The results indicated that 5.0% B-C3O exhibited stable catalytic ability at pH 3.0-9.0 and high degradation efficiency in co-existing inorganic Cl−, SO42−, NO3−, H2PO4− and organic humic acid. Non-radical 1O2, radicals •OH and SO4●−, as well as ClO− were detected with synergy effect for NOR degradation. By quantifying free radicals, •OH with 0.52 µM and SO4●− with 10.91 µM were obtained at 180 min, verifying the leading role of SO4●−. The degradation process involved the defluorination and decarboxylation, as well as opening of quinolone and piperazinyl rings. Adopting alfalfa as the model plant, the toxicity effect before and after NOR degradation was finally evaluated with seed germination rate and chlorophyll content as the physiological indicators. In summary, non-metal B-doping not only provides a creative strategy for the development of ferromagnetic perovskite-like materials, but also affords excellent catalysts for aiding the advanced oxidation technology for removal of emerging pollutants in wastewaters.

Thymidylate synthase (TS) is a key enzyme in the de novo biosynthesis of thymidine monophosphate, serving as a well-known drug target in chemotherapy against cancers and infectious diseases. Additional to its clinical value, TS is supposed to be a promising drug target in aquatic-disease control. To facilitate designing pathogen-specific TS inhibitors for shrimp-disease control, we report the crystal structures of TS from Litopenaeus vannamei (LvTS) in the apo form, LvTS-dUMP complex and LvTS-dUMP-raltitrexed complex at 2.27 A, 1.54 A, and 1.56 A resolution, respectively. LvTS shares a similar fold with known TSs, existing as a dimer in the crystal. The apo LvTS and LvTS-dUMP take an open conformation, and raltitrexed binding induces structural changes into a closed conformation in LvTS-dUMP-raltitrexed. Compared to those in other known TS-dUMP-raltitrexed complexes with the closed conformation, the C-terminal loop in LvTS-dUMP-raltitrexed shifts its position away from the bound raltitrexed; the distance between C6 of dUMP and Sγ of the catalytic cysteine is obviously longer than that in the known TS structures with closed conformations, resembling that in the TS structures with open conformations. Other species-specific interactions with dUMP and raltitrexed are also observed. Therefore, LvTS-dUMP-raltitrexed adopts a loosely closed conformation with structural features intermediate between the closed and the open conformations that were reported in other TSs. Our study provides the first crustcean TS structure, and reveals species-specific interactions between TSs and the ligands, which would facilitate designing pathogen-specific TS inhibitors for shrimp-disease control.

Hydrogen sulfide (H2S), a well-known signaling molecule, exerts significant regulatory effects on the cardiovascular and nervous systems. Therefore, monitoring the metabolism of H2S offers a potential mechanism to detect various diseases. In addition, biotin is significantly used as a targeting group to detect cancer cells exclusively. In this work, a biotin-guided benzoxadizole-based fluorescent probe, NP-biotin, was developed for H2S detection and evaluated in normal liver cell (LO2) and liver cancer cell (HepG2) lines. Results reveal that NP-biotin can detect cellular H2S with high sensitivity and selectivity. Moreover, NP-biotin has been confirmed to possess the ability to target cancer cells under the guidance of the biotin group.

dUTPases are key enzymes in all life kingdoms. A staphylococcal repressor protein (Stl) inhibited dUTPases from multiple species to various extents. Understanding the molecular basis underlying the inhibition differences is crucial to develop effective proteinaceous inhibitors of dUTPases. Herein, we report the complex structure of Stl N-terminal domain (StlN-ter) and Litopenaeus vannamei dUTPase domain (lvDUT65-210). Stl inhibited lvDUT65-210 through its N-terminal domain. The lvDUT65-210-StlN-ter complex structure revealed a heterohexamer encompassing three StlN-ter monomers bound to one lvDUT65-210 trimer, generating two types of Stl-dUTPase interfaces. Interface I is formed by Stl interaction with the lvDUT65-210 active-site region that is contributed by motifs I-IV from its two subunits; interface II results from Stl binding to the C-terminal motif V of the third lvDUT65-210 subunit. Structural comparison revealed both conserved features and obvious differences in Stl-dUTPase interaction patterns, giving clues about the inhibition differences of Stl on dUTPases. Noticeably, interface II is only observed in lvDUT65-210-StlN-ter. The Stl-interacting residues of lvDUT65-210 are conserved in other eukaryotic dUTPases, particularly human dUTPase. Altogether, our study presents the first structural model of Stl interaction with eukaryotic dUTPase, contributing to a more complete view of Stl inhibition and facilitating the development of proteinaceous inhibitor for eukaryotic dUTPases.

Extracellular proteases are often produced as pre-pro-enzyme and then undergo multiple processing steps to mature into the active form. The protease Epp, a virulent factor of Vibrio anguillarum, belongs to this family. Its maturation might be regulated by Ca2+ via its polycystic kidney disease (PKD) domain, but the molecular mechanism is unknown. Herein, we report the crystal structure of the first PKD domain from V. anguillarum Epp (Epp-PKD1) and its specific Ca2+ -binding capacity. Epp-PKD1 exists as a monomer, consisting of seven beta-strands which form two beta-sheets stacking with each other. One Ca2+ is bound by the residues Asn3, Gln4, Asp27, Asp29, Asp68 and a water molecule with a pentagonal bipyramidal geometry. Incubating the apo Epp-PKD1 with Ca2+ but not Mg2+, Mn2+, or Zn2+, enhances the thermal and chemical stability of Epp-PKD1, indicating its specific binding to Ca2+. Epp-PKD1 shares high similarity in both sequence and overall structure with that of Vibrio cholerae PrtV, a homologous protease of Epp, however, they differ in the oligomeric state and local structure at the Ca2+-binding site, suggesting maturation of PrtV and Epp might be differently regulated by Ca2+. Likely, proteases may take advantage of the structural diversity in PKD domains to tune their Ca2+-regulated maturation process. (C) 2018 Elsevier Inc. All rights reserved.

Semi-coke (SC) is a solid waste of coal chemical process, and the comprehensive utilization of SC is a challenging issue. Based on physicochemical characteristics, SC was firstly utilized as a mineral biochar for activating persulfate (PS) to promote simultaneous degradation of sulfadiazine (SDZ) and tetracycline (TC) in a binary mixture. The influencing parameters were systematically investigated. The dominant reactive oxygen species (ROS) were identified by combination of quenching experiments and electron spin resonance (EPR) analysis. Subsequently, possible activation mechanism and degradation pathways were also proposed. With the evolution of SDZ and TC in SC/PS system, the Vibrio fischeri was used to examine the acute toxicity to explore the detoxification efficiency. The experimental results indicated that in such a binary system, a mutually promoted degradation occurred between SDZ and TC, and at optimized conditions of 0.75 g/L PS, 0.50 g/L SC, pH 3.0 and 30 ℃, the developed SC/PS system had high degradation ability for simultaneous removal of SDZ (97.8%) and TC (97.2%). The polar groups and defects of SC were conducive to the adsorption; the oxygen-containing functional groups, pyridine N and graphite C on SC surface could be used as active sites for activating PS to produce ROS. During the degradation process, both radical (•OH and SO4•−) and non-radical (1O2) pathways coexisted. SC exhibited an excellent reusability, with the degradation efficiency of 84.5% for SDZ and 95.9% for TC after 4 cycles, and high mineralization efficiency of 67.5% after 4 h, demonstrating that such a unique SC/PS system exhibited a great potential for simultaneously efficient removal of different kinds of antibiotics in actual wastewaters.

Perfluorooctanoic acid (PFOA) is an anthropogenic persistent organic compound that resistant to traditional oxidative decomposition, bringing thus huge pressure to the environment. Here, the open hollow structured Calotropis gigantea fiber (CGF) was developed using a facile pyrolysis and subsequent magnetization method, and the resulting magnetic carbonized fiber (MC-CGF) was applied as an efficient catalyst for PFOA decomposition at room temperature in persulfate-based oxidation process. The experimental results indicated that with the addition of MC-CGF, the removal efficiency of PFOA could reach to 75.76% at 30℃ and pH 2.0. By calculation, the apparent activation energy was reduced from 62.76 kJ/mol to 22.49 kJ/mol, demonstrating that in the presence of MC-CGF, a lower reaction temperature and a shorter time were enough to reach the higher efficiency for PFOA decomposition. Mechanism research showed that during the decomposition process, PFOA underwent a first decarboxylation process and two possible decomposition pathways should also be taken into discussion. Taking pakchoi as the plant model, the process toxicity was finally evaluated, and the changes in chlorophyll content stated that the decomposition process of PFOA by MC-CGF assisted persulfate system was relatively harmless and environmental-friendly.

Although gonadotrophins are major regulators of ovarian function in teleosts and other vertebrates, accumulating evidence indicates that the growth hormone (GH)-insulin-like growth factor (IGF) axis also plays an important role in fish reproduction. As a first step to understand the physiological role of the GH-IGF system in the ovarian development of starry flounder (Platichthys stellatus), the expression profiles of GH and IGF messenger RNAs (mRNAs) and plasma GH, IGF-I, estradiol-17β (E2), and testosterone (T) levels during the ovarian development were investigated. The developmental stages of ovaries were divided into five stages (II, III, IV, V, and VI) by histological analysis. The hepatosomatic index (HSI) and gonadosomatic index (GSI) values increased and peaked at stage IV and stage V, respectively, and then declined at stage VI. Pituitary GH mRNA levels decreased sharply at stage III and raised to top level at stage VI. The hepatic IGF-I mRNA levels ascended to maximum value at stage V and then declined significantly at stage VI. However, the hepatic IGF-II mRNA levels remained stable and increased significantly at stage VI. In contrast, the ovarian IGF-I mRNA levels increased gradually and peaked at stage VI. The ovarian IGF-II mRNA levels were initially stable and increased significantly at stage V until the top level at stage VI. Consistent with the pituitary GH mRNA levels, plasma GH levels reduced sharply at stage III and remained depressed until stage V and then raised remarkably at stage VI. Plasma IGF-I level peaked at stage V and then declined to initial level. Plasma E2 level peaked at stage IV and then dramatically descended to the basal level. Plasma T level peaked at stage V and then declined significantly back to the basal level. Based on statistical analysis, significant positive correlations between hepatic IGF-I mRNA and GSI, ovarian IGF-II mRNA and hepatic IGF-II mRNA, ovarian IGF-I mRNA and ovarian IGF-II mRNA, and plasma IGF-I and plasma T were observed, respectively. These results suggest that the GH-IGF system may be involved in the ovarian development of starry flounder; GH and IGFs appear to play distinct roles in the regulation of the ovarian development in paracrine/autocrine manners. These findings extend our knowledge of the roles of the GH-IGF axis on reproduction regulation in fish.

Considering pH-dependent fluorescence of curcuminoids, a microemulsion electrokinetic chromatographic (MEEKC) method was developed under acidic conditions for their separation and detection using laser-induced native fluorescence (LINF), so as to solve the analysis of urine metabolism for curcuminoids. The microemulsion composition was optimized by response surface methodology (RSM), and the effects of buffer pH and organic modifiers were systematically investigated. The optimal buffer for the separation of curcuminoids was chosen as follows: 2.8% (v/v) ethyl acetate, 80 mM SDS and 2.8% (v/v) n-butanol to form microemulsion, 28% (v/v) ethanol as organic modifier, and 20 mM phosphoric acid as electrolyte at pH 3.0. Under these conditions, four curcuminoids including curcumin, demethoxy curcumin (DMC), bisdemethoxy curcumin (BDMC) and demethyl curcumin (DEC) could be well separated within 18 min, and the detection limits (LOD, based on S/N=3) were calculated to be 71, 60, 22, and 147 pg mL-1, respectively. Combined with solid-phase extraction (SPE), the developed MEEKC-LINF method has been successfully applied to continuously monitor the curcuminoids and related metabolites in human urine collected from a healthy volunteer after oral administration of curry, testifying that this method has potential for evaluating the pharmacological activity of curcuminoids.

The thymus is a lobulated unique lymphoid immune organ that plays a critical role in the selection, development, proliferation, and differentiation of T cells. The thymus of developing chickens undergoes continued morphological alterations; however, the biomolecular and transcriptional dynamics of the postnatal thymus in avian species is not clear yet. Therefore, the thymuses from chickens at different stages of development (at weeks 0, 1, 5, 9, 18, and 27) were used in the present study. The RNA-seq method was used to study the gene expression patterns. On average, 24120819 clean reads were mapped, differentially expressed genes (DEGs) were identified on the basis of log values (fold change), including 744 upregulated and 425 downregulated genes. The expression pattern revealed by RNA-seq was validated by quantitative real-time PCR (qPCR) analysis of four important genes, which are PCNA, CCNA2, CCNB2, and CDK1. Thus, the current study revealed that during postnatal development, the thymus undergoes severe atrophy. Thymus structure was damaged and gene expression changed dramatically, especially at the 27th week of age. Moreover, we found significant changes of several signaling pathways such as the cytokine-cytokine receptor interaction and cell cycle signaling pathways. Hence, it may be inferred that those signaling pathways might be closely related to the postnatal chicken thymus development.

The membrane progestin receptor α (mPRα) plays an important role in the regulation of oocyte meiotic maturation in fish. However, the endocrine regulation of multiple spawning in fish with asynchronous ovarian development remains to be determined. We identified four genes (FSHβ, LHβ, CGα and mPRα) that are widely expressed in both the reproductive and non-reproductive tissues of rock bream Oplegnathus fasciatus and examined their expression during ovarian maturation. The level of FSHβ transcripts in the pituitary remained high during vitellogenic stages, when the plasma levels of estradiol-17β reached maximum levels and oocyte development was stimulated. LHβ expression in the pituitary and ovary reached maximum levels during oocyte maturation when the plasma levels of testosterone also reached a maximum. A significant correlation between mPRα transcript levels and the gonadosomatic index was observed during vitellogenesis and final maturation. Brain gonadotropin (FSHβ and LHβ) and mPRα were continuously expressed at different stages of ovarian development. The dynamic and diverse nature of gonadotropins and mPRα gene expression patterns suggests that these genes might jointly regulate ovarian development and maturation through the brain–pituitary–ovary axis in the rock bream endocrine system. J. Exp. Zool. 323A: 430–444, 2015. © 2015 Wiley Periodicals, Inc.

Siderophore-interacting proteins (SIPs) play an important role in iron acquisition in many bacteria. SIPs release iron from the internalized ferric siderophore complex by reducing ferric iron to ferrous iron, but how the iron is reduced is not well understood. Here, asipgene was identified in the genome ofVibrio anguillarum775. To further understand the catalytic mechanism of the protein, the SIP was overexpressed inEscherichia coliRosetta (DE3) cells, purified and crystallized for X-ray diffraction analysis. The crystal diffracted to 1.113 Å resolution and belonged to space groupP21, with unit-cell parametersa = 64.63,b= 58.47,c= 70.65 Å, β = 114.19°.

In order to elucidate the possible roles of insulin-like growth factors I and II (IGF-I and IGF-II) in the embryonic development of Platichthys stellatus, their cDNAs were isolated and their spatial expression pattern in adult organs and temporal expression pattern throughout embryonic development were examined by quantitative real-time PCR assay. The IGF-I cDNA sequence was 1,268 bp in length and contained an open reading frame (ORF) of 558 bp, which encoded 185 amino acid residues. With respect to IGF-II, the full-length cDNA was 899 bp in length and contained a 648-bp ORF, which encoded 215 amino acid residues. The amino acid sequences of IGF-I and IGF-II exhibited high identities with their fish counterparts. The highest IGF-I mRNA level was found in the liver for both sexes, whereas the IGF-II gene was most abundantly expressed in female liver and male liver, gill, and brain. The sex-specific and spatial expression patterns of IGF-I and IGF-II mRNAs are thought to be related to the sexually dimorphic growth and development of starry flounder. Both IGF-I and IGF-II mRNAs were detected in unfertilized eggs, which indicated that IGF-I and IGF-II were parentally transmitted. Nineteen embryonic development stages were tested. IGF-I mRNA level remained high from unfertilized eggs to low blastula followed by a significant decrease at early gastrula and then maintained a lower level. In contrast, IGF-II mRNA level was low from unfertilized eggs to high blastula and peaked at low blastula followed by a gradual decrease. Moreover, higher levels of IGF-I mRNA than that of IGF-II were found from unfertilized eggs to high blastula, vice versa from low blastula to newly hatched larva, and the different expression pattern verified the differential roles of IGF-I and IGF-II in starry flounder embryonic development. These results could help in understanding the endocrine mechanism involved in the early development and growth of starry flounder.

Wetting behavior and interfacial characteristic of the Sn-3.5Ag/Ni system are investigated by sessile drop method at the temperature range of 523~773K. The reactive wetting processes demonstrate that: contact angles between the solder and Ni substrate decrease as exponential decay and the equilibrium contact angles decrease monotonously with the temperature increasing. Triple-line mobility is enhanced as the temperature increases. Compositions of the Sn-3.5Ag/Ni interface are identified by EPMA and EDS analysis as Ni3Sn4adjacent to the solder and Ni3Sn adjacent to the Ni substrate, respectively. The formation of the interface IMC was known to greatly improve the reliability of the solder joints in integrated circuits.

Wettability and interfacial characteristic of the Sn-3.0Ag-0.5Cu/Ni system are investigated by sessile drop method at the temperature range of 503~673K. The reactive wetting processes demonstrate that: contact angles between the solder and Ni substrate decrease as exponential decay and the equilibrium contact angles decrease monotonously with the temperature increasing. Triple-line mobility is enhanced as the temperature increases. Interface of the Sn-3.0Ag-0.5Cu /Ni interface are identified by EPMA and EDS analysis as (Cu,Ni)6Sn5 adjacent to the solder and Ni3Sn4 adjacent to the Ni substrate, respectively. Cu is condensed at the interface, the composition of (Cu,Ni)6Sn5 is (23.16~23.46)Ni- (36.56~37.52) Cu-(39.02~40.27)Sn (atom %). The formation of the (Cu,Ni)6Sn5 IMC was known to greatly improve the reliability of the solder joints in integrated circuits.

This study examined the effects of nicotinamide on proliferation, differentiation, and energy metabolism in a primary culture of bovine adipocytes. After treatment of cells with 100-500 μM nicotinamide, cell growth was measured using 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and cellular lipid content was assessed by Oil Red O staining and a triglyceride (TG) assay. Several factors related to energy metabolism, namely adenosine triphosphatase (ATPase) activity, nitric oxide (NO) content, nitric oxide synthase (NOS) activity, the number of mitochondria and the relative expression of glyceraldehydes-3- phosphate dehydrogenase (GAPDH), peroxisome proliferator-activated receptor-γ (PPARγ) and inducible NOS (iNOS), were also investigated. Results showed that nicotinamide induced both proliferation and differentiation in bovine preadipocytes. Nicotinamide decreased NO production by inhibiting NOS activity and iNOS mRNA expression, and controlled lipolytic activity by increasing ATPase activity and the number of mitochondria. The present study provides further evidence of the effects of nicotinamide on lipid and energy metabolism, and suggests that nicotinamide may play an important role in the development of bovine adipose tissue in vivo. This

It has been reported that there are multiple mechanisms by which bufalin could exert its antimetastatic effect. HIF-1α has been reported to be involved in tumor migration and invasion by regulating EMT. However, it is not known whether bufalin could exert the antimetastatic effect by modulating HIF-1α expression in hepatocellular carcinoma. In the present study, we aimed to evaluate the antimetastatic potential of bufalin in vivo and in vitro. Our results demonstrated that the liver/lung metastases were significantly reduced in bufalin-treated mice, as tested in the orthotopic transplanted and tail vein injection tumor models. Furthermore, the epithelial-to-mesenchymal transition (EMT) was inhibited in bufalin-treated tumors, as reflected the upregulation of E-cadherin, and downregulation of N-cadherin, vimentin, Snail. Similar results were observed in SMMC7721 cells treated with bufalin. Moreover, the transforming growth factor-β1 (TGF-β1)-induced EMT was also abrogated by bufalin. Mechanistically, our study demonstrated that hypoxia-inducible factor-1α (HIF-1α) played an important role in the antimetastatic effect of bufalin in hepatocellular carcinoma. Importantly, HIF-1α expression may be regulated through the inhibition of the PI3K/AKT/mTOR pathway. Taken together, our results suggest that bufalin suppresses hepatic tumor invasion and metastasis and that this process may be related to the PI3K/AKT/mTOR/ HIF-1α axis.

The membrane progestin receptor α (mPRα) plays an important role in the regulation of oocyte meiotic maturation in fish. However, the endocrine regulation of multiple spawning in fish with asynchronous ovarian development remains to be determined. We identified four genes (FSHβ, LHβ, CGα and mPRα) that are widely expressed in both the reproductive and non-reproductive tissues of rock bream Oplegnathus fasciatus and examined their expression during ovarian maturation. The level of FSHβ transcripts in the pituitary remained high during vitellogenic stages, when the plasma levels of estradiol-17β reached maximum levels and oocyte development was stimulated. LHβ expression in the pituitary and ovary reached maximum levels during oocyte maturation when the plasma levels of testosterone also reached a maximum. A significant correlation between mPRα transcript levels and the gonadosomatic index was observed during vitellogenesis and final maturation. Brain gonadotropin (FSHβ and LHβ) and mPRα were continuously expressed at different stages of ovarian development. The dynamic and diverse nature of gonadotropins and mPRα gene expression patterns suggests that these genes might jointly regulate ovarian development and maturation through the brain-pituitary-ovary axis in the rock bream endocrine system.

To evaluate the efficacy and safety of late-course hypofractionated radiation treatment of muscle-invasive bladder carcinoma after bladder-conserving surgery.Seventy-six patients with transitional cell bladder carcinoma, stage II (T2-4N0M0), after transurethral resection, were enrolled. Pirarubicin was given at 30 mg/mOne-, three- and five-year overall survival rates were 98, 78 and 69.5%, respectively. Mean survival time was 58.4 months (95% CI: 52.6, 64.2). In addition, 1-, 3- and 5-year local control rates were 100, 80.5 and 76.1%, respectively. Mean local control time was 60.7 months (95% CI: 55.1, 66.3). The cumulative incidence of local/regional failure and distant failure was 28.9%. The rate of single local/regional failure was 13.2%, but distant failure rate was 21.1%.Concurrent pirarubicin-based late-course hypofractionated radiation therapy showed desirable local control rate and acceptable toxicity. It could be used after bladder-conserving surgery to allow patients to preserve their bladder.

Obesity is characterized by excessive accumulation of white adipose tissue. Bromodomain-containing protein 2 (Brd2), which belongs to the bromodomain and extraterminal domain family of proteins, suppresses adipocyte differentiation. DNA methylation is critical for several differentiation processes and possibly in adipocyte differentiation. However, whether DNA methylation regulates the expression of Brd2 is not clear. In our study, we demonstrated that DNA methylation contributes to the regulation of Brd2 expression during pre-adipocyte differentiation. Brd2 mRNA levels were low in pre-adipocytes, increased in early adipocytes, and declined in mature adipocytes. To test whether and how Brd2 expression is regulated by DNA methylation during the differentiation of 3T3-L1 pre-adipocytes to adipocytes, cells were cultured in the presence of the methylation inhibitor 5-aza-2'-deoxycytidine (5-Aza). Pre-adipocytes and adipocytes exposed to 5-Aza exhibited a dose-dependent increase in Brd2 transcription levels, while only mature adipocytes exhibited increased expression of Brd2 protein. Subsequently, we tested the DNA methylation status of the Brd2 promoter region. Bisulfite-sequencing analysis revealed that six CpG sites in two predicted promoters of Brd2 were demethylated in early adipocytes and highly methylated in mature adipocytes. Digestion of bisulfite-converted PCR products of the Brd2 promoter region from 3T3-L1 cells with BstU1 (CGGC) revealed that the demethylation rate of the Brd2 promoter was consistent with Brd2 mRNA expression in differentiating 3T3-L1 cells. In conclusion, DNA demethylation of the Brd2 promoter region induced Brd2 expression during differentiation of 3T3-L1 cells into adipocytes.

In this paper, we present one method to develop and design an inexpensive electrolyzer simulation system, which can be applied to the management of the electrolyzer production. Firstly, we explore the framework of the electrolyzer simulation system. Our system is divided into several modules, including resource loading, scene rendering, scene interaction, collision detection and data visualization module. All these modules are designed particularly. Finally, we present the results and the results show that our method is feasible and valid. Our electrolyzer simulation system has been used in practical projects to manage the aluminum production and aluminum production education.