Alamandine attenuates hepatic fibrosis by regulating autophagy induced by NOX4-dependent ROS.

Angiotensin II (Ang II) has been reported to aggravate hepatic fibrosis by inducing NADPH oxidase (NOX)-dependent oxidative stress. Alamandine (ALA) protects against fibrosis by counteracting Ang II via the MAS-related G-protein coupled (MrgD) receptor, though the effects of alamandine on hepatic fibrosis remain unknown. Autophagy activated by reactive oxygen species (ROS) is a novel mechanism of hepatic fibrosis. However, whether autophagy is involved in the regulation of Ang II-induced hepatic fibrosis still requires investigation. We explored the effect of alamandine on hepatic fibrosis via regulation of autophagy by redox balance modulation. In vivo, alamandine reduced CCl4-induced hepatic fibrosis, hydrogen peroxide (H2O2) content, protein levels of NOX4 and autophagy impairment. In vitro, Ang II treatment elevated NOX4 protein expression and ROS production along with up-regulation of the angiotensin converting enzyme (ACE)/Ang II/Ang II type 1 receptor (AT1R) axis. These changes resulted in the accumulation of impaired autophagosomes in hepatic stellate cells (HSCs). Treatment with NOX4 inhibitor VAS2870, ROS scavenger N-acetylcysteine (NAC), and NOX4 small interfering RNA (siRNA) inhibited Ang II-induced autophagy and collagen synthesis. Alamandine shifted the balance of renin-angiotensin system (RAS) toward the angiotensin converting enzyme 2 (ACE2)/alamandine/MrgD axis, and inhibited both Ang II-induced ROS and autophagy activation, leading to attenuation of HSCs migration or collagen synthesis. In summary, alamandine attenuated liver fibrosis by regulating autophagy induced by NOX4-dependent ROS.

Angiotensin (1-7) inhibits arecoline-induced migration and collagen synthesis in human oral myofibroblasts via inhibiting NLRP3 inflammasome activation.

Arecoline induces oral submucous fibrosis (OSF) via promoting the reactive oxygen species (ROS). Angiotensin (1-7) (Ang-(1-7)) protects against fibrosis by counteracting angiotensin II (Ang-II) via the Mas receptor. However, the effects of Ang-(1-7) on OSF remain unknown. NOD-like receptors (NLRs) family pyrin domain containing 3 (NLRP3) inflammasome is identified as the novel mechanism of fibrosis. Whereas the effects of arecoline on NLRP3 inflammasome remain unclear. We aimed to explore the effect of Ang-(1-7) on NLRP3 inflammasome in human oral myofibroblasts. In vivo, activation of NLRP3 inflammasomes with an increase of Ang-II type 1 receptor (AT1R) protein level and ROS production in human oral fibrosis tissues. Ang-(1-7) improved arecoline-induced rats OSF, reduced protein levels of NADPH oxidase 4 (NOX4) and the NLRP3 inflammasome. In vitro, arecoline increased ROS along with upregulation of the angiotensin-converting enzyme (ACE)/Ang-II/AT1R axis and NLRP3 inflammasome/interleukin-1ß axis in human oral myofibroblasts, which were reduced by NOX4 inhibitor VAS2870, ROS scavenger N-acetylcysteine, and NOX4 small interfering RNA (siRNA). Furthermore, arecoline induced collagen synthesis or migration via the Smad or RhoA-ROCK pathway respectively, which could be inhibited by NLRP3 siRNA or caspase-1 blocker VX-765. Ang-(1-7) shifted the balance of RAS toward the ACE2/Ang-(1-7)/Mas axis, inhibited arecoline-induced ROS and NLRP3 inflammasome activation, leading to attenuation of migration or collagen synthesis. In summary, Ang-(1-7) attenuates arecoline-induced migration and collagen synthesis via inhibiting NLRP3 inflammasome in human oral myofibroblasts.

Metabolic engineering of Escherichia coli for enhanced production of D-pantothenic acid.

D-pantothenic acid (D-PA) is an essential vitamin that has been widely used in various industries. However, the low productivity caused by slow D-PA production in fermentation hinders its potential applications. In this study, strategies of engineering the synthetic pathway combined with regulating methyl recycle were employed in E. coli to enhance D-PA production. First, a self-induced promoter-mediated dynamic regulation of D-PA degradation pathway was carried out to improve D-PA accumulation. Then, to drive more carbon flux into D-PA synthesis, the key nodes of the R-pantoate pathway which encoded the essential enzyme were integrated into the genome. Subsequently, the further increase in D-PA production was achieved by promoting the regeneration of methyl donor. The strain L11T produced 86.03 g/L D-PA with a productivity of 0.797 g/L/h, which presented the highest D-PA titer and productivity to date. The strategies could be applied to constructing cell factories for producing other bio-based products.

A Study of Grain Selection in Two-Dimensional (2D) Grain Selectors during the Investment Casting of Single-Crystal Superalloy.

In this study, a series of Bridgman casting experiments were conducted to study the physical processes occurring in 2D grain selectors with different geometric parameters. The corresponding effects of the geometric parameters on grain selection were quantified by using an optical microscopy (OM) and a scanning electronic microscopy (SEM) equipped with electron backscatter diffraction (EBSD) function. Based on the results, the influences of the geometric parameters of the grain selectors are discussed, and an underlying mechanism accounting for the experimental results is proposed. The critical nucleation undercooling in the 2D grain selectors during grain selection was also analyzed.

Sex differences in antioxidant ability and energy metabolism level resulting in the difference of hypoxia tolerance in red swamp crayfish (Procambarus clarkii).

Sexual dimorphism widely exists in crustaceans. However, sex differences in the hypoxia tolerance of crayfish have rarely been reported. In this study, the differences in hypoxia tolerance between the two sexes of crayfish were assessed according to mortality, pathological features of hepatopancreas, antioxidant enzyme activity and differentially expressed genes (DEGs) analysis using transcriptome. The results showed that male crayfish displayed significantly higher mortality than the female under hypoxia stress (p < 0.05). Furthermore, female crayfish demonstrated higher levels of antioxidant enzyme activity. Hematoxylin-eosin staining analysis revealed that the damage of hepatopancreas was more severe in the male crayfish compared to the female crayfish. Additionally, there was higher expression level of the DEGs in hypoxia-inducible factor (HIF) pathway and higher energy metabolism level in the female compared to the male. Together, these findings suggest that the female crayfish with higher antioxidant ability and energy metabolism level exhibits stronger hypoxia tolerance than the male crayfish, providing the theoretical support for investigating sex differences in hypoxia tolerance among crustaceans.

Determination of Johnson-Cook Constitutive of 15-5 PH Steel Processed by Selective Laser Melting.

To obtain a Johnson-Cook model of 15-5 PH steel formed by selective laser melting (SLM), and to determine the difference between the forging process, in this work, mechanical testing, penetration testing and numerical simulations were used to study 15-5 PH steel formed by SLM and forging. Finally, the Johnson-Cook model parameters and failure parameters of the 15-5 PH steel formed by SLM and forging were obtained. We found that the SLM process was beneficial for refining the grain size of 15-5 PH steel and for improving the mechanical properties of 15-5 PH steel, where the yield strength of its specimens increased by 13.1% compared with the forged specimens. The error between the numerical simulations and penetration tests was less than 10%, which verified the validity of the numerical model parameters. It was also found that the penetration ability and abrasion resistance of the SLM-shaped projectiles were slightly superior to those of the forged projectiles.

Influence of Hf and MC Carbide on Transverse Platform in Single Crystal Grain Selection of 2D Grain Selector.

CM247LC Ni-based components have been widely used in developing hot ends in aero-engines and gas industrial turbines, and these have exhibited promising directional solidification (DS) results. However, the superalloy CM247LC shows defects after adding carbon (C) and hafnium (Hf). In this study, the effects of adding C and Hf on grain selection have been explored to enhance the 2D grain selector's performance and reduce casting costs. The experimental results reveal that the final region of carbide formation is where the dendrite is pushed into the paste region and finally solidifies. The performance requirements of carbide on the alloy can be controlled by changing the paste region and solidification sequence.

Split-Hopkinson Pressure Bar Test of Silicone Rubber: Considering Effects of Strain Rate and Temperature.

Using a split-Hopkinson pressure bar test instrument with a temperature control device, in this work, silicone rubber was tested at different temperatures (-40 °C-200 °C) and different strain rates (1.2 × 103 s-1-7.2 × 103 s-1). The results showed that the dynamic mechanical properties of silicone rubber were strain-rate sensitive at different temperatures and the yield strength of the silicone rubber increased with an increase in the strain rate. At a higher strain rate, silicone rubber showed temperature sensitivity. With a decrease in the strain rate, the influence of temperature on silicone rubber gradually decreased. Differential scanning calorimetry analysis showed that silicone rubber had good thermal stability at high temperatures. When the temperature was as low as -40 °C, the silicone rubber underwent a glass transition, showing the characteristics of brittle materials.

The first determination and analysis of the complete mitochondrial genome of Ancistrus temmincki (Siluriformes: Loricariidae).

In order to fully comprehend the evolution and kinship of fishes in the family of Loricariidae, the complete mitochondrial genome of the Loricariidae fish Ancistrus temmincki was firstly characterized in the present study. The whole mitogenome was 16,657 bp in size and consisted of 13 protein-coding genes, 22 tRNAs, 2 rRNAs genes, a control region and origin of light-strand replication. The proportion of coding sequences with a total length of 11,473 bp was 68.88%, which encoded 3,813 amino acids. The genome composition was highly A + T biased (56.29%), and exhibited AT-skew (0.0661) and a negative GC-skew (-0.2740). All protein-coding genes were started with ATG except for GTG in CO1, while stopped with the standard TAN codons or a single T. The control region (D-loop) ranging from 15,635 bp to 16,657 bp was 1023 bp in size. Until now, there is hardly any studies on the complete mitochondrial sequence in the genus of Ancistrus, phylogenetic analysis showed that A. temmincki was most closely related to Ancistrus cryptophthalmus in the genus of Ancistrus. The complete mitochondrial genome sequence has provided a new insight into the taxonomic classification, and a more complex picture of the species diversity within the family of Loricariidae.

Grain Orientation Optimization of Two-Dimensional Grain Selector during Directional Solidification of Ni-Based Superalloys.

The grain selection method is widely used in industry to produce Ni-based single crystal superalloys. A Z-form two-dimensional (2D) grain selector was designed to obtain high-quality single crystals. To control grain orientation deviation, one of the most important defects of the single crystal superalloys in casting, Z-form 2D grain selectors with different take-off angle were investigated in this study. The MM247LC superalloy single crystal samples were obtained by the Bridgman method modified by the Z-form grain selectors in this study. The Electron Backscattered Diffraction (EBSD) and the Optical Microscopy (OM) were used to observe and measure the grain selection growth and the microstructural evolution and orientation of the single crystal were also discussed. The results show that a Z-form 2D grain selector with an appropriate take-off angle can significantly reduce the deviation of the grain orientation. A single crystal superalloy with a deviation angle less than 6° can be obtained effectively when the take-off angle was 40°.

Proteomic analysis of extracellular vesicles released from heat-stroked hepatocytes reveals promotion of programmed cell death pathway.

Liver injury is a common complication of severe heat stroke (HS). Extracellular vesicles (EVs) are part of a novel pathway mediating intercellular communication. Whether EVs are involved in the pathogenesis underlying HS-induced liver injury remains unknown. Here, we explored the role of hepatocyte EVs in HS-induced liver injury and their protein regulation patterns after HS induction. Isobaric tags for relative and absolute quantification technology (iTRAQ) and liquid chromatography-tandem mass spectrometry analysis identified changes in the proteomic profiles of hepatocyte-derived heat-stroked EVs, and we identified 53 up-regulated proteins. Bioinformatics analysis verified that the regulation of programmed cell death was the most significant altered pathway. To clarify the effect of HS hepatocyte-derived EVs in inducing hepatocyte-programmed death and injury, they were added to recipient hepatocytes and injected into mice. This treatment significantly induced the synthesis of apoptosis (caspase-3/8) and necroptosis-associated proteins [receptor-interacting protein 1 (RIP1), RIP3, and mixed lineage kinase domain-like protein]; moreover, it increased the numbers of apoptotic and necroptotic cells in hepatocytes and liver tissues and increased the levels of biochemical liver injury markers (alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase). Our study is the first comprehensive analysis of the hepatocyte-derived heat-stroked EV proteome confirming the induction of liver injury by Evs. We provide a novel explanation for the pathological mechanism underlying HS-induced liver injury.

[Enhancing thermostability of xylanase from rumen microbiota by molecular cyclization].

The capacity for thermal tolerance is critical for industrial enzyme. In the past decade, great efforts have been made to endow wild-type enzymes with higher catalytic activity or thermostability using gene engineering and protein engineering strategies. In this study, a recently developed SpyTag/SpyCatcher system, mediated by isopeptide bond-ligation, was used to modify a rumen microbiota-derived xylanase XYN11-6 as cyclized and stable enzyme C-XYN11-6. After incubation at 60, 70 or 80 ℃ for 10 min, the residual activities of C-XYN11-6 were 81.53%, 73.98% or 64.41%, which were 1.48, 2.92 or 3.98-fold of linear enzyme L-XYN11-6, respectively. After exposure to 60-90°C for 10 min, the C-XYN11-6 remained as soluble in suspension, while L-XYN11-6 showed severely aggregation. Intrinsic and 8-anilino-1-naphthalenesulfonic acid (ANS)-binding fluorescence analysis revealed that C-XYN11-6 was more capable of maintaining its conformation during heat challenge, compared with L-XYN11-6. Interestingly, molecular cyclization also conferred C-XYN11-6 with improved resilience to 0.1-50 mmol/L Ca²âº or 0.1 mmol/L Cu²âº treatment. In summary, we generated a thermal- and ion-stable cyclized enzyme using SpyTag/SpyCatcher system, which will be of particular interest in engineering of enzymes for industrial application.

2-D Selector Simulation Studies on Grain Selection for Single Crystal Superalloy of CM247LC.

In the present work, the single crystal superalloy CM247LC was selected as the research material. By using directional experiments and the cellular automaton finite element (CAFE) model, the process of grain texture evolution in a two-dimensional grain selector was investigated to clarify the mechanism of grain selection in the two-dimensional passage during the process of directional solidification (DS). To optimize single crystal turbine blade production processes, the effects of grain selector geometries (i.e., selector diameter and pitch length, take-off angle) on the microstructure and stray grain were simulated and discussed.

Heatstroke-induced hepatocyte exosomes promote liver injury by activating the NOD-like receptor signaling pathway in mice.

BACKGROUND: Liver injury is a common and important clinical issue of severe heat stress (HS), which has toxic effects and promotes subsequent multiple organ failure. The pathogenesis of HS-induced liver injury has not been fully elucidated. Passively injured hepatocytes also drive liver injury. Exosomes, extracellular vesicles secreted by hepatocytes as "danger signals," mediate the intercellular transportation of diverse functional protein cargoes and modulate the biological processes of target cells. However, whether hepatocyte exosomes are involved in HS-induced liver injury has not been reported. The purpose of the current study was to clarify the release of hepatocyte exosomes under HS conditions and to explore their role in mediating HS-induced liver injury. METHODS: HS was induced in hepatocytes or mice by hyperthermic treatment at 43.0 °C for 1 h. Exosomes from control and HS-exposed hepatocytes were isolated by standard differential ultracentrifugation. The hepatocyte exosomes were characterized, and the differentially expressed proteins of the control and HS exosomes were identified by isobaric tags for relative and absolute quantitation (iTRAQ) mass spectrometry and subjected to Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. Recipient hepatocytes were treated with control or HS exosomes, whereas in vivo, the exosomes were infused into mice. The internalization of HS hepatocyte exosomes by hepatocytes or the liver was tracked. The effect of HS exosomes on the activation of the NOD-like receptor signaling pathway and liver injury was demonstrated in vitro and in vivo. RESULTS: HS induced an increase in the release of exosomes from hepatocytes, which were internalized by recipient liver cells in vitro and taken up by the liver in vivo. HS significantly changed the proteomic profiles of hepatocyte exosomes based on the iTRAQ analysis. The KEGG pathway analysis revealed the enrichment of proteins associated with injury and inflammatory signaling pathways, especially the NOD-like receptor signaling pathway, the activity of which was upregulated. Subsequently, the capacity of HS hepatocyte exosomes to activate the NOD-like receptor signaling pathway was verified and found to aggrevate liver damage and inflammation in vitro and in vivo. CONCLUSIONS: This study is the first preliminary study to demonstrate the induction of acute liver injury by hepatic exosomes in the setting of severe HS and reveals potentially related pathways. These results provide a basis for future research and the identification of new targets for clinical intervention.

Development of a High-Efficiency Z-Form Selector for Single Crystal Blades and Corresponding Grain Selection Mechanism.

Single crystal (SX) is widely used in modern turbine blades to improve the creep fracture, fatigue, oxidation, and coating properties of the turbine, so that the turbine engine has excellent performance and durability. In this paper, the single crystal super alloy MM247LC is used as the research material. The evolution of grain structure in a two-dimensional grain selector was studied by directional experiments, and the mechanism of grain selection in the two-dimensional channel during directional solidification was clarified. In order to optimize the production process of single crystal turbine blades, the effects of the geometrical structure of a Z-type separator (i.e., wire diameter and take-off angle) on the crystal orientation, microstructure, and grain efficiency of blades were discussed.

Grain Selection in a High-Efficiency 2D Grain Selector During Casting of Single-Crystal Superalloys.

Using electron backscattered diffraction techniques (EBSD) and optical microscopy (OM), the grain selection and competitive growth in a new-designed high-efficiency two-dimensional (2D) selector during solidification of Ni-based single-crystal (SX) superalloys have been investigated with emphasis on the geometry of the selector part in this article. It is found that the efficiency of the grain selector depends greatly on the thickness and eccentric distance of the selector part. When the thickness is smaller than 3 mm, a single grain can be selected. After reducing this value, the grain selector becomes more effective. When the eccentric distance is larger than 8 mm, one grain can be selected. As the eccentric distance increases, the selector's efficiency is optimized. Recommendations for optimizing the geometry of the selector part are provided.

Using a Three-Dimensional Reduction Method in the High-Efficiency Grain Selector and Corresponding Grain Selection Mechanism.

In the development of a high-efficiency grain selector, the spiral selectors are widely used in Ni-based single crystal (SX) superalloys casting to produce single crystal turbine blades. For the complex three-dimensional structure of the spiral, a 2D grain selector was designed to investigate in this paper. As a result, the parameters of two-dimensional grain selection bond and the corresponding grain selection mechanism were established, and the three-dimensional grain selection bond was designed again by means of two-dimensional coupling optimization parameters.

Investigation of Start Block and Corresponding Influence for Grain Selection during Casting of Single-Crystal Superalloys.

To figure out the impact of the parameters of a starter block (the diameter D and height H) on grain selection and the selecting mechanism, a spiral selector was measured with optical microscopy (OM) and electron backscatter diffraction (EBSD) during the solidification of Ni-based single crystal (SX) superalloys. In this experiment, starter blocks with diameters of 8 mm, 10 mm, 15 mm, and 30 mm and a height of 30 mm were designed to find the best parameters. Recommendations for optimizing starter block geometry are provided.

AVE 0991 Attenuates Pyroptosis and Liver Damage after Heatstroke by Inhibiting the ROS-NLRP3 Inflammatory Signalling Pathway.

We previously demonstrated that angiotensin-(1-7) (Ang-(1-7)), an essential endocrine factor, inhibits the NLRP3 inflammasome by regulating reactive oxygen species (ROS) in fibrotic livers. We also demonstrated that the NLRP3 inflammasome contributes to the liver damage induced by pyroptosis after heatstroke. However, the role of Ang-(1-7) in the hepatocytes under heat stress remains uncertain. We aimed to examine the change in angiotensin peptides in the livers affected by heatstroke and the effect on the ROS-NLRP3 inflammatory signalling pathway. In vivo, increased angiotensin II (Ang II) and decreased Ang-(1-7) in the serum of heatstroke patients suffering from hepatic dysfunction were observed. The change in angiotensin peptides was considered a potential biomarker that could be used to predict hepatic dysfunction. Enhanced Ang II and attenuated Ang-(1-7) levels were also observed in the liver tissue of heatstroke rats, which were consistent with their receptors and converting enzymes. Hepatic damage associated with increased ROS and protein expression levels of NOX4, NLRP3, caspase-1, and IL-1ß was attenuated by AVE 0991, an analogue of Ang-(1-7). In vitro, pyroptosis, characterized by activated caspase-1 and IL-1ß, was observed in hepatocytes under heat stress, which was enhanced by Ang II and attenuated by antioxidants, NOX4 siRNA, and AVE 0991. In summary, AVE 0991 attenuates pyroptosis and liver damage induced by heat stress by inhibiting the ROS-NLRP3 inflammatory signalling pathway.