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Based on brake noise dynamometer test data, combined with the artificial intelligent algorithms, frictional braking noise is quantitatively analyzed and predicted in this study. To achieve this goal, a frictional braking noise prediction method is indicatively proposed, which consists of two main parts: first, based on the experimental data obtained from the brake noise dynamometer tests, and combining with the improved Long-Short-Term Memory (LSTM) algorithm, the coefficients of friction (COFs) are predicted under various braking test conditions. Then, based on the predicted braking COFs and other selected critical braking parameters, the quantitative prediction of frictional braking noise is obtained by means of the optimized eXtreme Gradient Boosting (XGBoost) algorithm. Finally, the inherent features of the XGBoost algorithm are employed to qualitatively analyze the importance of the main factors affecting the frictional braking noise. The prediction algorithms of COFs and frictional braking noise are validated by the brake dynamomter test data, and the R2 (R square) scores of both the LSTM and XGBoost prediction algorithms are 0.9, which verifies the feasibility of both algorithms. The main contribution of this work is to predict the braking noise based on a large set of test data and combined with the LSTM and XGBoost artificial intelligent algorithms, which can significantly save time for the brake system development and braking performance testing, and has significance to the rapid prediction of braking frictional noise and fast NVH (noise, vibration, and harshness) optimal design of frictional braking systems.

Background Elevated homocysteine is epidemiologically related to insulin resistance. Protein-tyrosine phosphatase 1B (PTP1B) is a negative regulator of insulin signaling. However, the effect of homocysteine on PTP1B remains unclear. Methods S-homocysteinylated PTP1B was identified by LC-ESI-MS/MS. The ability of thioredoxin system to recover active PTP1B from S-homocysteinylated PTP1B was confirmed by RNA interference. To address the mechanism for homocysteine to affect PTP1B activity, we performed 5-IAF insertion, activity assays, Western blotting, co-immunoprecipitation and glucose uptake experiments. Results The thiol-containing form of homocysteine (HcySH) suppressed phosphorylation of insulin receptor-β subunit, but enhanced PTP1B activity. This phenomenon was partially related to the fact that HcySH promoted PTP1B expression. Although the disulfide-bonded form of homocysteine (HSSH) modified PTP1B to form an inactive S-homocysteinylated PTP1B, HcySH-induced increase in the activities of cellular thioredoxin and thioredoxin reductase, components of thioredoxin system, could recover active PTP1B from S-homocysteinylated PTP1B. Thioredoxin system transferred electrons from NADPH to S-homocysteinylated PTP1B, regenerating active PTP1B in vitro and in hepatocytes. The actions of HcySH were also related with decrease in hepatic glucose uptake. Conclusions The effect of HcySH/HSSH on PTP1B activity depends, at least partially, on the ratio of active PTP1B and S-homocysteinylated PTP1B. High HcySH-induced an increase in thioredoxin system activity is beneficial to de-S-homocysteinylation and is good for PTP1B activity. General significance Our data provide a novel insight into post-translational regulation of PTP1B, and expand the biological functions of thioredoxin system.

Acute exacerbation of chronic obstructive pulmonary disease (AECOPD) is a global problem with high mortality. Its pathogenesis is not fully understood. To reveal new serum feature of AECOPD and their potential implications, we have analyzed 180 serum samples, and found that in the serum of AECOPD patients, 4-hydroxy-2-nonenal (4HNE)-protein adducts are dynamically increased as partial pressure of oxygen (PaO2) drops, which is accompanied by progressively decreasing thioredoxin reductase (TrxR1) and thioredoxin (Trx1), as compared with those of healthy people. This phenomenon is unique, because acute hypoxia patients have 1.1-fold or 1.7-fold higher serum TrxR1 or Trx1 activity, respectively, than healthy people, in keeping with low 4HNE level. Moreover, serum 4HNE-protein adducts may form disulfide-linked complexes with high-molecular-weight, the amount of which is significantly increased during AECOPD. Serum 4HNE-protein adducts include 4HNE-Trx1 adduct and 4HNE-TrxR1 adduct, but only the former is significantly increased during AECOPD. Through cell biology, biochemistry and proteomics methods, we have demonstrated that extracellular 4HNE and 4HNE-Trx1 adduct affect human bronchial epithelial cellsviadifferent mechanisms. 4HNE-Trx1 adduct may significantly alter the expression of proteins involved mainly in RNA metabolism, but it has no effect on TrxR1/Trx1 expression and cell viability. On the other hand, low levels of 4HNE promote TrxR1/Trx1 expression and cell viability, while high levels of 4HNE inhibit TrxR1/Trx1 expression and cell viability, during which Trx1, at least in part, mediate the 4HNE action. Our data suggest that increasing serum 4HNE and decreasing serum Trx1 in AECOPD patients are closely related to the pathological processes of the disease. This finding also provides a new basis for AECOPD patients to use antioxidant drugs.

Background Serum thioredoxin of type-2 diabetic patients is significantly higher than that of healthy people. Pathophysiological significance is unclear. Methods Effects of serum/extracellular thioredoxin on phosphorylation (activation) of hepatic insulin receptor (IR) were investigated by using methods in biochemistry, cell/molecular biology and mass spectrometry. Results In human serum, thioredoxin and insulin may interact. Their mixture contains a mixed disulfide between insulin B-chain and thioredoxin-Cys73, which limits their activities. In contrast, free form of serum/extracellular thioredoxin is active, and can regulate phosphorylation of insulin receptor β-subunits (IRβ) via direct/indirect mechanisms. The direct mechanism associates with positive regulation. Serum/extracellular thioredoxin increases insulin binding to IR, facilitating insulin-induced phosphorylation of IRβ and downstream AKT. The indirect mechanism is involved in negative regulation. Entry of extracellular thioredoxin into hepatic cells via IR enhances the expression and activity of cellular protein-tyrosine phosphatase 1B (PTP1B), which negatively regulates IRβ phosphorylation. After coordination between these two mechanisms, the positive impact of serum/extracellular thioredoxin overwhelms its negative impact on IRβ phosphorylation, which subsequently accelerates hepatic glucose uptake. In hepatic cells with thioredoxin deficiency, insulin-induced IRβ phosphorylation is decreased, which could be restored by extracellular thioredoxin entry. Moreover, the results from assaying 475 serum samples demonstrate a discriminating value of serum thioredoxin activity in diagnosing type-2 diabetes. Conclusion Serum/extracellular thioredoxin plays a critical role in regulating hepatic IRβ phosphorylation. General significance In case of insulin resistance/type-2 diabetes, hepatic IRβ is at low phosphorylation level, thereby the improvement effect of serum/extracellular thioredoxin on insulin-induced IRβ phosphorylation seems particularly important.

Thioredoxin is a redox protein while adiponectin is an adipokine. Their relationship has been less appreciated. Here we show that in serum of patients with acute exacerbations of chronic obstructive Pulmonary Disease, decreased activity of thioredoxin coexists with increased level of adiponectin as partial pressure of arterial oxygen decreases. From the results with adiponectin-knockout mice and in vitro experiments, we have found a reciprocal inhibitory relationship, by which adiponectin inhibits cytosolic thioredoxin (Trx1) expression, whereas serum Trx1 influences adiponectin multimerization. The association between extracellular Trx1 and adiponectin attenuates their functions. This relationship is dynamic, and correlated with a body’s physiological conditions.

Background and purpose Insulin-sensitizing drugs are currently limited, and identifying new candidates is a challenge. Protein tyrosine phosphatase 1B (PTP1B) negatively regulates insulin signalling, and its inhibition is anticipated to improve insulin resistance. Here, the pharmacological properties of CX08005, a novel PTP1B inhibitor, were investigated. Experimental approach Recombinant hPTP1B protein was used to study enzyme activity and mode of inhibition. Docking simulation explored the interactions between CX08005 and PTP1B. Insulin sensitivity was evaluated by glucose tolerance test (GTT) in diet-induced obese (DIO) and KKAy mice; glucose-stimulated insulin secretion (GSIS), homeostasis model assessment of insulin resistance index (HOMA-IR) and whole-body insulin sensitivity (ISWB ) were also determined. A hyperinsulinaemic-euglycaemic clamp was performed to evaluate insulin-stimulated glucose disposal in both whole-body and insulin-sensitive tissues. Furthermore, CX08005's effects on muscle, fat and liver cells were determined in vitro. Key results CX08005 competitively inhibited PTP1B by binding to the catalytic P-loop through hydrogen bonds. In DIO mice, CX08005 ameliorated glucose intolerance dose-dependently (50-200 mg·kg(-1) ·day(-1) ) and decreased the HOMA-IR. In KKAy mice, CX08005 (50 mg·kg(-1) ·day(-1) ) improved glucose intolerance, GSIS, ISWB and hyperglycaemia. CX08005 also enhanced insulin-stimulated glucose disposal, increased glucose infusion rate and glucose uptake in muscle and fat in DIO mice (hyperinsulinaemic-euglycaemic clamp test). CX08005 enhanced insulin-induced glucose uptake in 3T3-L1 adipocytes and C2C12 myotubes, and increased phosphorylation of IRβ/IRS1 and downstream molecules in hepatocytes in a dose- and insulin-dependent manner respectively. Conclusions and implications Our results strongly suggest that CX08005 directly enhances insulin action in vitro and in vivo through competitive inhibition of PTP1B.

The simulation software of manipulator is generally developed by manufacturers, and the products are only suitable for the specific company, but cannot be used in universal. Aiming at this problem, this paper establishes a mathematical model with attributes through parametric design. The standard D-H parameter method is used to model the kinematics of the manipulator. Based on the expert system, the intelligent path planning of the manipulator is carried out, and the trajectory control instructions of the manipulator are generated. According to the Monte Carlo method of point cloud, the workspace of manipulator is obtained, and the bounding box technology is used for real-time collision detection. Taking the welding work as an example, the parametric modelling and trajectory planning simulation of the manipulator are realized, and a general off-line programming platform is established.

High level of palmitate is associated with metabolic disorders. We recently showed that enhanced level of S-palmitoylated cytosolic thioredoxin (Trx1) in mouse liver was new characteristic feature of insulin resistance. However, our understanding of the effect of S-palmitoylation on Trx1 is limited, and the tissue specificity of Trx1 S-palmitoylation is unclear. Here we show that S-palmitoylation also occurs at Cys73 of Trx1 in living endothelial cells, and the level of S-palmitoylated Trx1 undergoes regulation by insulin signaling. Trx1 prefers thiol-thioester exchange with palmitoyl-CoA to acetyl-CoA. S-palmitoylation alters conformation or secondary structure of Trx1, as well as decreases the ability of Trx1 to transfer electrons from thioredoxin reductase to S-nitrosylated protein-tyrosine phosphatase 1B and S-nitroso-glutathione. Our results demonstrate that S-palmitoylation is an important post-translational modification of human Trx1.

Zhong, L. and Li, X., 2015. Simulation analysis of lightweight cylindrical lattice materials with different unit cells. To address the difficulty of designing lattice material structures, five different unit cells are presented including edge structure, vertex structure, face center structure, interconnect vertex structure and cross face center structure. The finite element analysis (FEA) method is introduced to analyze the mechanical properties of these five typical unit cells. It is indicated that, when stressed, bended or twisted with the same shape, size, mass, and loading force by changing their cellular pillar section R, the edge structure possesses optimal performance. The simulation results of the functional efficiency and bearing efficiency data obtained for various unit cells could shed some light on the lattice material designing under different loading conditions.

Overnutrition can lead to oxidative stress, but its underlying mechanism remains unclear. In this study, we report that human liver-derived HepG2 cells utilize cytosolic thioredoxin reductase (TrxR1) and thioredoxin (hTrx1) to defend against the high glucose/palmitate-mediated increase in reactive oxygen species. However, enhanced TrxR1/hTrx1 palmitoylation occurs in parallel with a decrease in their activities under the conditions studied here. An autoacylation process appears to be the major mechanism for generating palmitoylated TrxR1/Trx1 in HepG2 cells. A novel feature of this post-translational modification is the covalent inhibition of TrxR1/hTrx1 by palmitoyl-CoA, an activated form of palmitate. The palmitoyl-CoA/TrxR1 reaction is NADPH-dependent and produces palmitoylated TrxR1 at an active site selenocysteine residue. Conversely, S-palmitoylation occurs at the structural Cys62/Cys69/Cys72 residues but not the active site Cys32/Cys35 residues of hTrx1. Palmitoyl-CoA concentration and the period of incubation with TrxR1/hTrx1 are important factors that influence the inhibitory efficacy of palmitoyl-CoA on TrxR1/hTrx1. Thus, an increase in TrxR1/hTrx1 palmitoylation could be a potential consequence of high glucose/palmitate. The time-dependent inactivation of the NADPH-TrxR1-Trx1 system by palmitoyl-CoA occurs in a biphasic manner - a fast phase followed by a slow phase. Kinetic analysis suggests that the fast phase is consistent with a fast and reversible association between TrxR1/hTrx1 and palmitoyl-CoA. The slow phase is correlated with a slow and irreversible inactivation, in which selenolate/thiolate groups nucleophilically attack the α-carbon of bound palmitoyl-CoA, leading to the formation of thioester/selenoester bonds. hTrx1 can enhance rate of fast phase but limits the rate of slow phase when it is present in a preincubation mixture containing NADPH, TrxR1 and palmitoyl-CoA. Therefore, hTrx1 may provide palmitoylation sites or partially protect the TrxR1 active site selenol/thiol group(s) from palmitoylation. Our data suggest that Se/S-palmitoylation acts as an important modulator of TrxR1/hTrx1 activities, representing a novel potential mechanism that underlies overnutrition-induced events.

Abnormally enhanced tissue factor (TF) activity is related to increased thrombosis risk in which oxidative stress plays a critical role. Human cytosolic thioredoxin (hTrx1) and thioredoxin reductase (TrxR), also secreted into circulation, have the power to protect against oxidative stress. However, the relationship between hTrx1/TrxR and TF remains unknown. Here we show reversible association of hTrx1 with TF in human serum and plasma samples. The association is dependent on hTrx1-Cys-73 that bridges TF-Cys-209 via a disulfide bond. hTrx1-Cys-73 is absolutely required for hTrx1 to interfere with FVIIa binding to purified and cell-surface TF, consequently suppressing TF-dependent procoagulant activity and proteinase-activated receptor-2 activation. Moreover, hTrx1/TrxR plays an important role in sensing the alterations of NADPH/NADP+ states and transducing this redox-sensitive signal into changes in TF activity. With NADPH, hTrx1/TrxR readily facilitates the reduction of TF, causing a decrease in TF activity, whereas with NADP+, hTrx1/TrxR promotes the oxidation of TF, leading to an increase in TF activity. By comparison, TF is more likely to favor the reduction by hTrx1-TrxR-NADPH. This reversible reduction-oxidation reaction occurs in the TF extracellular domain that contains partially opened Cys-49/-57 and Cys-186/-209 disulfide bonds. The cell-surface TF procoagulant activity is significantly increased after hTrx1-knockdown. The response of cell-surface TF procoagulant activity to H2O2 is efficiently suppressed through elevating cellular TrxR activity via selenium supplementation. Our data provide a novel mechanism for redox regulation of TF activity. By modifying Cys residues or regulating Cys redox states in TF extracellular domain, hTrx1/TrxR function as a safeguard against inappropriate TF activity. Background: Oxidant-induced elevation in tissue factor activity increases thrombosis risk. Results: Tissue factor can form a disulfide bridge with human cytosolic thioredoxin and be reduced by thioredoxin-thioredoxin reductase-NADPH along with a decrease in its activity. Conclusion: Thioredoxin/thioredoxin reductase is involved in orchestration of tissue factor-dependent events. Significance: Revealing novel modulators can be exploited for therapeutic benefits in tissue factor-related disorders.

A novel immunoassay using the eggshell membrane as immobilization platform for determination of human IgG (HIgG) has been developed. The immunoassay was based on a sandwich immunoreaction of rabbit anti-human IgG (primary antibody), HIgG and the goat anti-human IgG labeled with FITC (secondary antibody). Due to well permeability and highly biocompatibility of eggshell membrane, a friendly microenvironment was constructed to prolong the life-times of the immobilized proteins. Under the optimal conditions, the fluorescence intensity is linear to the concentration of HIgG in a range of 20–100 ng mL−1 (r = 0.9918). The proposed method was successfully applied to the determination of the HIgG in the standard blood serum, and the results are satisfactory compared with that obtained by general immunonephelometric method.

(−)-Epigallocatechin gallate (EGCG) is a major bioactive component in leaves of green tea, and has been widely investigated for its anti-tumor activity. The interaction between EGCG and the key peptides or proteins (e.g. glutathione, enzymes) in vivo is thought to be involved in the toxicity and anti-cancer mechanism of EGCG. However, the true anti-tumor mechanism of EGCG is not clear, and few studies have focused on the reactivity of EGCG toward peptides or proteins under physiological conditions (pH 7.4, 37°C). In this work, the covalent interactions between EGCG and model peptides containing one or more nucleophilic residues (i.e. Arg, Cys, Met, and α-NH2 of the N-terminus of peptides) under physiological condition were fully characterized using mass spectrometry. It was found that EGCG can react with the thiol groups of peptides to form adducts under physiological conditions (pH 7.4, 37°C), even in the absence of the peroxidase/hydrogen peroxide system. Besides the thiol groups of peptides, it is firstly reported that EGCG also reacts with α-NH2 of the N-terminus or arginine residues of model peptides to form Schiff base adducts, and the methionine residues of model peptides can be easily oxidized by hydrogen peroxide (H2O2) generated during the process of EGCG auto-oxidation to form methionine sulfoxide products. The preference for the reaction of nucleophlic residues of peptides with EGCG was determined to have the following order: Cys > α-NH2 of the N-terminus > Arg. The neutral loss ions of [M+H–170]+ and [M+H-138]+ were detected in all tandem mass spectra of the EGCG adducts of peptides, which indicates that these two neutral loss ions can be considered as the characteristic neutral loss ions of peptides modified by EGCG. Results of the present research provide insights into the toxicology and anti-tumor mechanism of EGCG in vivo. Copyright © 2009 John Wiley & Sons, Ltd.

Epigallocatechin 3-gallate (EGCG) is the most abundant catechin in green tea and may combat bacteria with few side-effects. Its selectivity for different bacterial infections remains unclear, and hence the identification of the underlying mechanism is of practical importance. Both the thioredoxin (Trx) system and the glutathione/glutaredoxin (Grx) system support bacterial growth. Some pathogenic bacteria are naturally deficient in the Grx system. We analyzed the effect of green tea extract (GTE) and EGCG on wild-type and null mutants of Escherichia coli with either Trx or Grx system deficiency and found that GTE and EGCG selected the Trx system as a target and killed the mutant that is exclusively dependent on Trx/Trx reductase (TrxR). EGCG inhibited the activity of both Trx1 and TrxR of E. coli in a dose-dependent and time-dependent manner. The IC50 values of EGCG for the reduced forms of E. coli Trx1/TrxR were ~ 3-4-fold lower than those for their non-reduced forms. The IC50 value of EGCG for the E. coli Trx1 system was 56-fold lower than that for the mammalian Trx1 system. The inhibition by EGCG of both Trx1 and TrxR of E. coli was irreversible. EGCG-induced inactivation of E. coli Trx1 was a second-order process, and that of E. coli TrxR was an affinity-labeling process. The covalent binding sites for EGCG in E. coli Trx1 were Trp(28) , Trp(31) and Cys(32) , and in E. coli TrxR were Cys(135) and Cys(138) . Moreover, the sensitivity of Staphylococcus aureus to EGCG was similar to that of an E. coli mutant with Grx system deficiency. EGCG-induced inactivation of Trx/TrxR in S. aureus coincided with suppressed growth of this virulent pathogen. Our findings suggest a role for EGCG-dependent Trx/TrxR inactivation in potentiating antibacterial activity of EGCG.

Insulin-sensitizing drugs are currently limited, and identifying new candidates is a challenge. Protein tyrosine phosphatase 1B (PTP1B) negatively regulates insulin signalling, and its inhibition is anticipated to improve insulin resistance. Here, the pharmacological properties of CX08005, a novel PTP1B inhibitor, were investigated.Recombinant hPTP1B protein was used to study enzyme activity and mode of inhibition. Docking simulation explored the interactions between CX08005 and PTP1B. Insulin sensitivity was evaluated by glucose tolerance test (GTT) in diet-induced obese (DIO) and KKAy mice; glucose-stimulated insulin secretion (GSIS), homeostasis model assessment of insulin resistance index (HOMA-IR) and whole-body insulin sensitivity (ISWB ) were also determined. A hyperinsulinaemic-euglycaemic clamp was performed to evaluate insulin-stimulated glucose disposal in both whole-body and insulin-sensitive tissues. Furthermore, CX08005's effects on muscle, fat and liver cells were determined in vitro.CX08005 competitively inhibited PTP1B by binding to the catalytic P-loop through hydrogen bonds. In DIO mice, CX08005 ameliorated glucose intolerance dose-dependently (50-200 mg·kg(-1) ·day(-1) ) and decreased the HOMA-IR. In KKAy mice, CX08005 (50 mg·kg(-1) ·day(-1) ) improved glucose intolerance, GSIS, ISWB and hyperglycaemia. CX08005 also enhanced insulin-stimulated glucose disposal, increased glucose infusion rate and glucose uptake in muscle and fat in DIO mice (hyperinsulinaemic-euglycaemic clamp test). CX08005 enhanced insulin-induced glucose uptake in 3T3-L1 adipocytes and C2C12 myotubes, and increased phosphorylation of IRβ/IRS1 and downstream molecules in hepatocytes in a dose- and insulin-dependent manner respectively.Our results strongly suggest that CX08005 directly enhances insulin action in vitro and in vivo through competitive inhibition of PTP1B.

Thioredoxin reductases (TrxRs) from mammalian cells contain an essential selenocysteine residue in the conserved C-terminal sequence Gly-Cys-SeCys-Gly forming a selenenylsulfide in the oxidized enzyme. Reduction by NADPH generates a selenolthiol, which is the active site in reduction of Trx. The three-dimensional structure of the SeCys498Cys mutant of rat TrxR in complex with NADP + has been determined to 3.0-Å resolution by x-ray crystallography. The overall structure is similar to that of glutathione reductase (GR), including conserved amino acid residues binding the cofactors FAD and NADPH. Surprisingly, all residues directly interacting with the substrate glutathione disulfide in GR are conserved despite the failure of glutathione disulfide to act as a substrate for TrxR. The 16-residue C-terminal tail, which is unique to mammalian TrxR, folds in such a way that it can approach the active site disulfide of the other subunit in the dimer. A model of the complex of TrxR with Trx suggests that electron transfer from NADPH to the disulfide of the substrate is possible without large conformational changes. The C-terminal extension typical of mammalian TrxRs has two functions: ( i ) it extends the electron transport chain from the catalytic disulfide to the enzyme surface, where it can react with Trx, and ( ii ) it prevents the enzyme from acting as a GR by blocking the redox-active disulfide. Our results suggest that mammalian TrxR evolved from the GR scaffold rather than from its prokaryotic counterpart. This evolutionary switch renders cell growth dependent on selenium.

We have determined the sequence of 23 peptides from bovine thioredoxin reductase covering 364 amino acid residues. The result was used to identify a rat cDNA clone (2.19 kilobase pairs), which contained an open reading frame of 1496 base pairs encoding a protein with 498 residues. The bovine and rat thioredoxin reductase sequences revealed a close homology to glutathione reductase including the conserved active site sequence (Cys-Val-Asn-Val-Gly-Cys). This also confirmed the identity of a previously published putative human thioredoxin reductase cDNA clone. Moreover, one peptide of the bovine enzyme contained a selenocysteine residue in the motif Gly-Cys-SeCys-Gly (where SeCys represents selenocysteine). This motif was conserved at the carboxyl terminus of the rat and human enzymes, provided that TGA in the sequence GGC TGC TGA GGT TAA, being identical in both cDNA clones, is translated as selenocysteine and that TAA confers termination of translation. The 3'-untranslated region of both cDNA clones contained a selenocysteine insertion sequence that may form potential stem loop structures typical of eukaryotic selenocysteine insertion sequence elements required for the decoding of UGA as selenocysteine. Carboxypeptidase Y treatment of bovine thioredoxin reductase after reduction by NADPH released selenocysteine from the enzyme with a concomitant loss of enzyme activity measured as reduction of thioredoxin or 5,5'-dithiobis(2-nitrobenzoic acid). This showed that the carboxyl-terminal motif was essential for the catalytic activity of the enzyme.

Long-term treatment with ambroxol (ABX), a bronchial expectorant, was found to prevent acute exacerbation of chronic obstructive pulmonary disease (AECOPD). The underlying mechanism remains unclear. To address this, we have investigated the effect of ABX on critical antioxidant proteins thioredoxin (Trx) and thioredoxin reductase (TrxR) that are decreased in patients with AECOPD. Trx, TrxR and NADP(H) form Trx system, which is involved in regulating numerous oxidative stress-related events. In human bronchial epithelial cells, treatment with ABX from 0 to 200 μM gradually increased mRNA and protein levels of TrxR/Trx. At these ABX concentrations, TrxR activity was elevated progressively, whereas Trx activity exhibited a dose-dependent biphasic response, increasing at 50 and 75 μM, but decreasing at ABX over 150 μM. Pre-treatment with 75 μM ABX enhanced the capacity of the cells to eliminate reactive oxygen species, which was largely prevented by knockdown of cytosolic Trx (hTrx1). In a purified system, ABX shortened the initial lag phase during the reduction of insulin disulfide by Trx system. Pre-treatment of NADPH-reduced TrxR with ABX caused a dose- and time-dependent increase in thiolate/selenolate species, i.e. the catalytically active form of TrxR. Kinetic analysis demonstrated that the reduction of H2O2 by TrxR or Trx system were enhanced by 100 or 200 μM ABX. When hTrx1 was mixed with ABX in a molar ratio of 1:1 to 1:100 (which could occur in human plasma), changes in intrinsic Trp fluorescence occurred, and the response of reduced hTrx1 was especially remarkable. These data reveal an ABX-sensing mechanism of TrxR/Trx. We therefore conclude that the antioxidant actions of ABX at physiological concentrations are, at least partially, mediated by TrxR and/or Trx system.

Abdominal trauma accounts for nearly 20% of all severe traffic injuries and can often result from intentional physical violence, from which blunt liver injury is regarded as the most common result and is associated with a high mortality rate. Liver injury may be caused by a direct impact with a certain velocity and energy on the abdomen, which may result in a lacerated liver by penetration of fractured ribs. However, liver ruptures without rib cage fractures were found in autopsies in a series of cases. All the victims sustained punches on the abdomen by fist. Many studies have been dedicated to determining the mechanism underlying hepatic injury following abdominal trauma, but most have been empirical. The actual process and biomechanism of liver injury induced by blunt impact on the abdomen, especially with intact ribs remained, are still inexhaustive. In order to investigate this, finite element methods and numerical simulation technology were used. A finite element human torso model was developed from high resolution CT data. The model consists of geometrically-detailed liver and rib cage models and simplified models of soft tissues, thoracic and abdominal organs. Then, the torso model was used in simulations in which the right hypochondrium was punched by a fist from the frontal, lateral, and rear directions, and in each direction with several impact velocities. Overall, the results showed that liver rupture was primarily caused by a direct strike of the ribs induced by blunt impact to the abdomen. Among three impact directions, a lateral impact was most likely to cause liver injury with a minimum punch speed of 5 m/s (the momentum was about 2.447 kg.m/s). Liver injuries could occur in isolation and were not accompanied by rib fractures due to different material characteristics and injury tolerance.

The elucidation and prediction of the biomechanics of lower limb fractures could serve as a useful tool in forensic practices. Finite element (FE) analysis could potentially help in the understanding of the fracture mechanisms of lower limb fractures frequently caused by car-pedestrian accidents. Our aim was (1) to develop and validate a FE model of the human lower limb, (2) to assess the biomechanics of specific injuries concerning run-over and impact loading conditions, and (3) to reconstruct one real car-pedestrian collision case using the model created in this study. We developed a novel lower limb FE model and simulated three different loading scenarios. The geometry of the model was reconstructed using Mimics 13.0 based on computed tomography (CT) scans from an actual traffic accident. The material properties were based upon a synthesis of data found in published literature. The FE model validation and injury reconstruction were conducted using the LS-DYNA code. The FE model was validated by a comparison of the simulation results of three-point bending, overall lateral impact tests and published postmortem human surrogate (PMHS) results. Simulated loading scenarios of running-over the thigh with a wheel, the impact on the upper leg, and impact on the lower thigh were conducted with velocities of 10 m/s, 20 m/s, and 40 m/s, respectively. We compared the injuries resulting from one actual case with the simulated results in order to explore the possible fracture bio-mechanism. The peak fracture forces, maximum bending moments, and energy lost ratio exhibited no significant differences between the FE simulations and the literature data. Under simulated run-over conditions, the segmental fracture pattern was formed and the femur fracture patterns and mechanisms were consistent with the actual injury features of the case. Our study demonstrated that this simulation method could potentially be effective in identifying forensic cases and exploring of the injury mechanisms of lower limb fractures encountered due to inflicted lesions. This model can also help to distinguish between possible and impossible scenarios.

On the basis of studying the structure and design specification of Flender’s gear units, a method of parametric design and modeling for batch gear units based on Pro/Engineer has been adopted. According to given dimensions, we has initially defined variables in Pro/Engineer and established a family table which comprises all the required dimensions, then assigned variables or equations to corresponding feature dimensions during modeling. The model of a gear unit consists of two sections: one is the fundamental shape, the dimensions of which are all given; the other is the external embellish, including the bosses around the axes, the oil gallery for lubrication, the inspecting hole and so on, the dimensions of which are to be designed and determined by designers so as to create an ideal figure of the case. Thus, each series of gear units merely requires a single model, so that all parts in the entire series of gear units shall be generated through the family table and saved as an individual product respectively. The experimental results demonstrate that the development period is shortened, the cost is reduced and the productivity is increased.

Thioredoxin (Trx) and thioredoxin reductase (TrxR) function as antioxidant and anti-apoptotic proteins, which are often up-regulated in drug-resistant cancer cells. (-)-epigallocatechin-3-gallate (EGCG) is a naturally occurring antioxidant in green tea, but also exhibits prooxidant and apoptosis-inducing properties. We have previously showed a linkage between EGCG-induced inactivation of TrxR and decreased cell survival, revealing TrxR as a new target of EGCG. However, the molecular events underlying the importance of Trx/TrxR in EGCG-induced cytotoxicity remain unclear. Here, we show that the crosstalk between EGCG and Trx/TrxR occurred in a redox-dependent manner, and EGCG induced inactivation of Trx/TrxR in parallel with increased ROS levels in HeLa cells. Moreover, EGCG displayed great reactivity with Cys/Sec residues that have low pK(a) values. The structure of EGCG suggests that its quinone form would readily react with thiolate and selenolate nucleophiles. Using mass spectrometry, we have demonstrated the formation of EGCG-Trx1 (Cys(32)) and EGCG-TrxR (Cys/Sec) conjugates, confirming that EGCG quinone specifically conjugates with active-site Cys(32) in Trx or C-terminal Cys/Selenocysteine (Sec) couple in TrxR under conditions where Trx/TrxR are reduced. Non-reduced form of Trx/TrxR could escape from EGCG inhibition. These data reveal a potential mechanism for enhancing EGCG-induced cancer cell death by the NADPH-dependent reduction of Trx/TrxR.

Complex network theory is put forward in following research to analyze the statistical parameters of network business statically in E-commerce area. Empirical Study is based on the data provided by an online shopping website and statistical parameters such as enterprise act degree and clustering coefficient is studied accordingly. Thus we found that E-commerce displays the power law and clustering as most complex networks. It’s verified that the effect of customer nodes with high degree, so called as those customers with high loyalty, is important to develop new customer resources. Therefore we come to a conclusion that complex network theory is a feasible and effective tool for E-commerce company to deploy their business with the research of customer loyalty.

Objective: Elevation of homocysteine and thioredoxin (Trx) levels was found in some patients with coronary artery diseases (CAD). However, their correlations with CAD were not clear. Dysfunction of thioredoxin/thioredoxin reductase (TrxR) may cause oxidative stress that is common to CAD. We seek to determine the association among homocysteine, Trx/TrxR and CAD. Methods: Serum samples were collected from 150 CAD patients under statin treatment and 122 nonCAD controls. Risk factors for atherosclerosis including homocysteine, lipids and glucose levels were analyzed. Trx/TrxR activities and protein levels were determined using super-insulin assay and Western blot, respectively. One-way ANOVA, Tukey’s post hoc test and Spearman’s rank correlation coefficient were used for statistical analysis. CAD severity was evaluated by angiographic Gensini score. Results: Compared with non-CAD group, CAD group had significantly increased TrxR activity (P < 0.05) and homocysteine levels (P < 0.01), but not Trx activity. After further dividing CAD group using homocysteine below 15M as reference, Trx activity decreased significantly in CAD group with high homocysteine, and was inversely associated with homocysteine levels (r = −0.199, P < 0.05) that was, however, weakly positively associated with TrxR activity. Neither lipids nor glucose significantly affected Trx/TrxR activity. Association of CAD severity with low Trx plus high homocysteine was strong (r = −0.458, P < 0.001), but with high homocysteine alone was rather weak (r = 0.125, P = 0.225). Conclusion: In CAD patients, high homocysteine levels may cause low Trx activity, which is closely correlated to the extent and severity of CAD.

Mammalian cytosolic thioredoxin reductase is a homodimer of 55 kDa subunit containing an essential penultimate selenocysteine residue. An active analogue of the rat enzyme in which cysteine replaces selenocysteine has been expressed in Escherichia coli cells at high levels and purified to homogeneity. The pure enzyme contains one FAD per subunit and shows spectral properties identical to that of the wild-type thioredoxin reductase. The isolated enzyme in its oxidized and reduced forms or the enzyme complexed with NADP(+) was crystallized by the hanging-drop vapour-diffusion method. The diffraction pattern extends to 3 A resolution. The crystals are monoclinic, space group P2(1), with unit-cell parameters a = 78.9, b = 140.5, c = 170.8 A, alpha = 94.6 degrees. There are three dimeric molecules in the asymmetric unit.

The material requisition flow in the warehousing management was studied in the article, and combining the mining algorithm of causal rules, the material requisition based on data mining was developed. The system could fulfill the management requirements of the material requisition process for the enterprise, mine the data of material return and deeply analyze reasons, which can help the enterprise to enhance the production quality and reduce the production cost.

Elevated blood histamine plays a role in the pathogenesis of atherosclerosis. Calcium signaling mediates histamine action in endothelial cells. Selenium (Se) is a dietary essential trace element for humans. Se compounds in different oxidation states were found to exhibit an opposing effect on the histamine-induced calcium signaling in the ECV304 cell line. When Se in the form of sodium selenite was added in the cell culture, the reactivity of the histamine H(1)-receptor was increased as reported in our previous paper. We here show that as a culture supplement, sodium selenite enhanced the activity of selenoprotein thioredoxin reductase (TrxR) and the calcium response to histamine stimulation, which were reversed by treating the cells with gold thioglucose, a nucleophilic drug that selectively modifies thiolate/selenolate groups. Sodium selenite most likely caused a reductive shift in the thiol/disulfide redox balance through increasing TrxR activity. In contrast, when the cells were treated with Se in the form of ebselen, a thiol oxidant with peroxidase-like activity, histamine-induced calcium release and calcium entry were significantly suppressed. This effect appeared related to the thiol-directed modification rather than the peroxidase-like activity of ebselen, because this inhibitory effect was not replicated by increasing cellular peroxidase activity. Thus, the opposing effects of sodium selenite and ebselen on histamine-induced calcium signaling are achieved, at least in part, through their opposite actions in modulating the thiol/disulfide redox state.

Selenium is an essential micronutrient for humans and animals, and its deficiency can predispose to the development of pathological conditions. This study evaluates the effect of selenium deficiency on the thioredoxin system, consisting of NADPH, selenoprotein thioredoxin reductase (TrxR), and thioredoxin (Trx); and the glutathione system, including NADPH, glutathione reductase, glutathione, and glutaredoxin coupled with selenoprotein glutathione peroxidase (GPx). We particularly investigate whether inactive truncated TrxR is present under selenium-starvation conditions due to reading of the UGA codon as stop. Feeding rats a selenium-deficient diet resulted in a large decrease in activity of TrxR and GPx in rat liver but not in the levels of Trx1 and Grx1. However, selenium deficiency induced mitochondrial Grx2 10-fold and markedly changed the expression of some flavoproteins that are involved in the cellular folate, glucose, and lipid metabolism. Liver TrxR mRNA was nearly unchanged, but no truncated enzyme was found. Instead, a low-activity form of TrxR with a cysteine substituted for the penultimate selenocysteine in the C-terminal active site was identified in selenium-deficient rat liver. These results show a novel mechanism for decoding the UGA stop codon, inserting cysteine to make a full-length enzyme that may be required for selenium assimilation.—Lu, J., Zhong, L., Lönn, M. E., Burk, R. F., Hill, K. E., Holmgren, A. Penultimate selenocysteine residue replaced by cysteine in thioredoxin reductase from selenium-deficient rat liver.

(-)-Epigallocatechin gallate (EGCG) is a major bioactive component in leaves of green tea, and has been widely investigated for its anti-tumor activity. The interaction between EGCG and the key peptides or proteins (e.g. glutathione, enzymes) in vivo is thought to be involved in the toxicity and anti-cancer mechanism of EGCG. However, the true anti-tumor mechanism of EGCG is not clear, and few studies have focused on the reactivity of EGCG toward peptides or proteins under physiological conditions (pH 7.4, 37 degrees C). In this work, the covalent interactions between EGCG and model peptides containing one or more nucleophilic residues (i.e. Arg, Cys, Met, and alpha-NH(2) of the N-terminus of peptides) under physiological condition were fully characterized using mass spectrometry. It was found that EGCG can react with the thiol groups of peptides to form adducts under physiological conditions (pH 7.4, 37 degrees C), even in the absence of the peroxidase/hydrogen peroxide system. Besides the thiol groups of peptides, it is firstly reported that EGCG also reacts with alpha-NH(2) of the N-terminus or arginine residues of model peptides to form Schiff base adducts, and the methionine residues of model peptides can be easily oxidized by hydrogen peroxide (H(2)O(2)) generated during the process of EGCG auto-oxidation to form methionine sulfoxide products. The preference for the reaction of nucleophlic residues of peptides with EGCG was determined to have the following order: Cysalpha-NH(2) of the N-terminusArg. The neutral loss ions of [M+H-170](+) and [M+H-138](+) were detected in all tandem mass spectra of the EGCG adducts of peptides, which indicates that these two neutral loss ions can be considered as the characteristic neutral loss ions of peptides modified by EGCG. Results of the present research provide insights into the toxicology and anti-tumor mechanism of EGCG in vivo.

Applying to the system engineering, software engineering theory and methods, a system is designed with subjective and objective, qualitative and quantitative evaluation and intelligent decision support. Analytic Hierarchy Process (AHP) makes the weight setting more reasonable. Meanwhile enterprise’s comprehensive evaluation model has been constructed by the efficiency coefficient method with multi-objective decision-making. In the system, the expandable decision-making knowledge database was designed in the form of single-tree inference, which can give feasible decision-making suggestions based on different evaluation result.

Mammalian cytosolic thioredoxin reductase (TrxR1) is an attractive target for developing cancer chemopreventive agents since its inhibition is associated with a reduced growth of cancer cells. However, the known inhibitors of this enzyme mostly have a toxic effect on human health. We report on a non-toxic inhibitor, green tea. TrxR1 was found to be inhibited by green tea extracts (Gte) with an IC50 value of 256 microg/ml. Catechins, the major components of Gte, showed various inhibitory effects, in which (-)-epigallocatechin-3-gallate (EGCG) exhibited a stronger inhibition than any other catechins tested. The inhibition of TrxR1 by EGCG was close to competitive (Ki = 64 microM) with substrate DTNB and was non-competitive (Ki = 92 microM) with co-enzyme NADPH. The preincubation of TrxR1 with EGCG led to irreversible enzyme inactivation in a time-dependent manner, which was highly effective in the presence of NADPH. The inactivation included an equilibrium step used to form a reversible TrxR1-EGCG complex (EI) (dissociation constant Ki* = 43 microM), and an isomerization step used to form an irreversible complex (E*I) (rate constant k3 = 4.8 x 10(-3) s(-1)). We have identified thiol/selenol groups in the active site as reactive sites that mediated TrxR1 inhibition by EGCG. When cultured HeLa cells were treated with Gte or EGCG for 22-24 h, TrxR1 activity in cell extracts was significantly inhibited, accompanied by a reduction of cell viability in a concentration-dependent manner (IC50 = 40 microg/ml for Gte and 107 microM for EGCG). The inactivation of TrxR1 by Gte/EGCG is most likely linked to a reduction of HeLa cell viability.

The technique of three dimensional solid element model and assembly was used to determine the temperature field of radial-flow impeller. The FEM software Cosmos was applied to analyze model system, and the precise analytic results were obtained. The results show that the analytic model can reflect the stead-state and transient temperature field characteristics of impeller directly, and thus can be worthy reference to analyze and calculate the temperature field of impeller in engineering design.

Black tea is recently reported to have anti-carcinogenic effects through pro-oxidant property, but the underlying mechanisms remain unclear. Mammalian cytosolic thioredoxin reductase (TrxR1) is well -known for its anti-oxidation activity. In this study, we found that black tea extract (BTE) and theaflavins (TFs), the major black tea polyphenols, inhibited the purified TrxR1 with IC(50) 44 microg/ml and 21+/-1 microg/ml, respectively. Kinetics of TFs exhibited a mixed type of competitive and non-competitive inhibition, with K(is) 4+/-1 microg/ml and K(ii) 26+/-5 microg/ml against coenzyme NADPH, and with K(is) 12+/-3 microg/ml and K(ii) 27+/-5 microg/ml against substrate DTNB. In addition, TFs inhibited TrxR1 in a time-dependent manner. In an equilibrium step, a reversible TrxR1-TFs complex (E*I) forms, which is followed by a slow irreversible first-order inactivation step. Rate constant of the inactivation was 0.7 min(-1), and dissociation constant of E*I was 51.9 microg/ml. Treatment of NADPH-reduced TrxR1 with TFs decreased 5-(Iodoacetamido) fluorescein incorporation, a fluorescent thiol-reactive reagent, suggesting that Sec/Cys residue(s) in the active site may be involved in the binding of TFs. The inhibitory capacity of TFs depends on their structure. Among the TFs tested, gallated forms had strong inhibitory effects. The interactions between TFs and TrxR1 were investigated by molecular docking, which revealed important features of the binding mechanism of theaflavins. An inhibitory effect of BTE on viability of HeLa cells was observed with IC(50) 29 microg/ml. At 33 microg/ml of BTE, TrxR1 activity in HeLa cells was decreased by 73% at 22 h after BTE treatment. TFs inhibited cell viability with IC(50) 10+/-4 microg/ml for HeLa cells and with IC(50) 20+/-5 microg/ml for EAhy926 cells. The cell susceptibility to TFs was inversely correlated to cellular levels of TrxR1. The inhibitory actions of TFs on TrxR1 may be an important mechanism of their anti-cancer properties.

The ways to set up document management system are proposed. These ways based on Java B/S adopts N-tier frameworkof MVC Model 2 in J2EE platform and uses EJB, Struts Web Framework and Hibernate technology and so on. The framework of this system and its functional modules are studied, which perfect and improve expansibility, robustness, loaded capabil-ity and executive efficiency. A basis for improving the quality of knowledge management and achieving data share and cooperative design based on Internet will be provided by this research for enterprises.

Intracellular Ca 2þ signaling controls many cellular functions. Understanding its regulation by selenoproteins is essential for understanding the role of selenoproteins in regulating cell functions. The activity of thioredoxin reductase (TrxR), thioredoxin (Trx) content, and the activity of glutathione peroxidase (GPx) in the human endothelial cells cultured in selenium-supplemented medium (refer as Se þ cells) was found 70%, 40%, and 20% higher, respectively than those in the cells cultured in normal medium (refer as Se 0 cells). The intracellular Ca 2þ signaling initiated by inositol 1,4,5-trisphosphate (IP3), histamine, thapsigargin (TG), carbonyl cyanide p-(tri-fluoromethoxy) phenyl-hydrazone (FCCP), and cyclosporin A (CsA) was investigated in both Se þ and Se 0 cells. It was interestingly found that the higher activity of selenoproteins reduced the sensitivity of IP3 receptor to the IP3-triggered Ca 2þ release from intracellular stores, but enhanced activation of the receptor-coupled phospholipase C in histamine-stimulated Se þ cells by showing much more generation of IP3 and higher elevation of cytosolic Ca 2þ . The higher selenoprotein activity also reduced susceptibility of the uniporter to the mitochondrial uncoupler, susceptibility of the permeability transition pore (PTP) to its inhibitor, and the vulnerability of endoplasmic reticulum (ER) Ca 2þ -ATPase to its inhibitor in selenium-supplementing cells. The results suggest that cell calcium signaling is subjected to thiol-redox regulation by selenoproteins. J. Cell. Physiol. 205: 97‐106

The thioredoxin system, composed of thioredoxin (Trx), thioredoxin reductase (TrxR), and NADPH, is a powerful protein disulfide reductase system with a broad substrate specificity. Recently the selenazol drug ebselen was shown to be a substrate for both mammalian TrxR and Trx. We examined if alpha-tocopherol quinone (TQ), a product of alpha-tocopherol oxidation, is reduced by ebselen in the presence of TrxR, since TQ was not a substrate for the enzyme itself. Ebselen reduction of TQ in the presence of TrxR was caused by ebselen selenol, generated from fast reduction of ebselen by the enzyme. TQ has no intrinsic antioxidant activity, while the product of reduction of TQ, alpha-tocopherolhydroquinone (TQH(2)), is a potent antioxidant. The thioredoxin system dependence of ebselen to catalyze reduction of other oxidized species, such as hydrogen peroxide, dehydroascorbate, and peroxynitrite, is discussed. The ability of ebselen to reduce TQ via the thioredoxin system is a novel mechanism to explain the effects of the drug as an antioxidant in vivo.

Mammalian thioredoxin reductases (TrxR) are dimers homologous to glutathione reductase with a selenocysteine (SeCys) residue in the conserved C-terminal sequence -Gly-Cys-SeCys-Gly. We removed the selenocysteine insertion sequence in the rat gene, and we changed the SeCys(498) encoded by TGA to Cys or Ser by mutagenesis. The truncated protein having the C-terminal SeCys-Gly dipeptide deleted, expected in selenium deficiency, was also engineered. All three mutant enzymes were overexpressed in Escherichia coli and purified to homogeneity with 1 mol of FAD per monomeric subunit. Anaerobic titrations with NADPH rapidly generated the A(540 nm) absorbance resulting from the thiolate-flavin charge transfer complex characteristic of mammalian TrxR. However, only the SeCys(498) --> Cys enzyme showed catalytic activity in reduction of thioredoxin, with a 100-fold lower k(cat) and a 10-fold lower K(m) compared with the wild type rat enzyme. The pH optimum of the SeCys(498) --> Cys mutant enzyme was 9 as opposed to 7 for the wild type TrxR, strongly suggesting involvement of the low pK(a) SeCys selenol in the enzyme mechanism. Whereas H(2)O(2) was a substrate for the wild type enzyme, all mutant enzymes lacked hydroperoxidase activity. Thus selenium is required for the catalytic activities of TrxR explaining the essential role of this trace element in cell growth.