The response of physiological and xylem anatomical traits under cadmium stress in Pinus thunbergii seedlings.

Studying the response of physiological and xylem anatomical traits under cadmium (Cd) stress is helpful to understand plants' response to heavy metal stress. Here, seedlings of Pinus thunbergii were treated with 50, 100 and 150 mg/kg Cd2+ for 28 days. Cd and nonstructural carbohydrate (NSC) content of leaves, stems and roots, root Cd2+ flux, Cd distribution pattern in stem xylem and phloem, stem xylem hydraulic traits, cell wall component fractions of stems and roots, phytohormonal content such as abscisic acid (ABA), gibberellic acid 3 (GA3), molecule -indole-3-acetic acid (IAA) and jasmonic acid (JA) from both leaves and roots, as well as xylem anatomical traits from both stems and roots were measured. Root Cd2+ flux increased from 50 to 100 mmol/L Cd2+ stress, however, decreased at 150 mmol/LCd2+. Cellulose and hemicellulose in leaves, stems and roots did not change significantly under Cd stress, while pectin decreased significantly. The NSC content of both leaves and stems showed significant changes under Cd stress while the root NSC content was not affected. In both leaves and roots, the ABA content significantly increased under Cd stress, while the GA3, IAA and JA-ME content significantly decreased. Both xylem hydraulic conductivity and xylem water potential decreased with Cd stress, however, tracheid diameter and double wall thickness of the stems and roots were not affected. High Cd intensity was found in both the stem xylem and phloem in all Cd stressed treatments. Our study highlighted the in-situ observation of Cd distribution in both the xylem and phloem, and demonstrated the instant response of physiological traits such as xylem water potential, xylem hydraulic conductivity, root Cd2+ flux, NSC content, as well as phytohormonal content under Cd stress, and the less affected traits such as xylem anatomical traits, cellulose and hemicellulose.

Kinetic Limits of Graphite Anode for Fast-Charging Lithium-Ion Batteries.

Fast-charging lithium-ion batteries are highly required, especially in reducing the mileage anxiety of the widespread electric vehicles. One of the biggest bottlenecks lies in the sluggish kinetics of the Li+ intercalation into the graphite anode; slow intercalation will lead to lithium metal plating, severe side reactions, and safety concerns. The premise to solve these problems is to fully understand the reaction pathways and rate-determining steps of graphite during fast Li+ intercalation. Herein, we compare the Li+ diffusion through the graphite particle, interface, and electrode, uncover the structure of the lithiated graphite at high current densities, and correlate them with the reaction kinetics and electrochemical performances. It is found that the rate-determining steps are highly dependent on the particle size, interphase property, and electrode configuration. Insufficient Li+ diffusion leads to high polarization, incomplete intercalation, and the coexistence of several staging structures. Interfacial Li+ diffusion and electrode transportation are the main rate-determining steps if the particle size is less than 10 µm. The former is highly dependent on the electrolyte chemistry and can be enhanced by constructing a fluorinated interphase. Our findings enrich the understanding of the graphite structural evolution during rapid Li+ intercalation, decipher the bottleneck for the sluggish reaction kinetics, and provide strategic guidelines to boost the fast-charging performance of graphite anode.

Enhanced adsorption of Congo red from urea/calcium chloride co-modified biochar: Performance, mechanisms and toxicity assessment.

Adsorbents with excellent physicochemical properties and green synthetic routes are desired for efficient removal of Congo red (CR) wastewater. Hence, a novel approach was proposed within this work. Biochar NCBC obtained from Medulla Tetrapanacis was synthesized through co-modification with urea/calcium chloride. NCBC exhibited an enormous surface area (750.09 m2/g) and a micro-mesoporous composite structure. Higher nitrogen content was detected on the surface of NCBC (8.17%) compared to that of urea directly modified biochar (4.63%). Nitrogen observed on the surface of NCBC was presented as graphitic N, pyrrolic N, amine N as well as pyridinic N. Kinetic and isothermal investigations revealed the active sites on NCBC to be homogeneous and bind to CR mainly by chemisorption. Calculated maximum sorption of CR on NCBC was 2512.82 mg/g basing on Langmuir model. Moreover, the practicality of NCBC was further proved by the cultivation of Nelumbo nucifera Gaertn. and Penicillium.

Efficient removal of Congo red and methylene blue using biochar from Medulla Tetrapanacis modified by potassium carbonate.

Biochar (BC) prepared from Medulla Tetrapanacis was recognized as having potential in environmental remediation because of its porous texture, abundant chemical surface groups and mineral composition. In this study, a novel modified biochar (KBC) derived from Medulla Tetrapanacis achieved efficient adsorption of Congo red (CR) and methylene blue (MB). Characterization results suggested that the modification had little effect on the group distribution of KBC, but a dramatic expansion of specific surface area was observed on KBC (1180.45 m2/g) compared to BC (198.51 m2/g). KBC exhibited a maximum sorption of 584.17 mg/g for CR and 318.01 mg/g for MB. Kinetic and isotherm studies revealed sorption of CR and MB by KBC was chemosorption, which occurred on the monolayer surface. The comprehensive analysis also provided the basis for the application of KBC to practical production. These outcomes suggested that KBC may become a new option for the effluent treatment.

Electron beam irradiation grafting of metal-organic frameworks onto cotton to prepare antimicrobial textiles.

Textiles modified with antimicrobial nanomaterials have excellent comprehensive performance. However, the shedding of nanoparticles often occurs in actual use. This not only reduces the service life of antimicrobial textiles, but also causes potential harm. Here, we report a new method to covalently immobilize a zinc-imidazolate MOF (ZIF-8) onto cotton fabric by electron beam irradiation to prepare antimicrobial textiles with excellent durability. A series of characterization analysis showed the electron beam irradiation did not damage the structure of the ZIF-8 nanoparticles and the particles were successfully introduced onto cotton fibers via poly hydroxyethyl acrylate (PHEA). The modified cotton fabric exhibited >99% inhibition of Escherichia coli, Staphylococcus aureus and Candida albicans. The results of dry cleaning and rub resistance tests showed that the prepared antimicrobial cotton fabric had significant durability which was attributed to the strong covalent binding between the MOF and textile.

3D Porous Structure-Inspired Lignocellulosic Biosorbent of Medulla tetrapanacis for Efficient Adsorption of Cationic Dyes.

The focus of this work was on developing a green, low-cost, and efficient biosorbent based on the biological structure and properties of MT and applying it to the remediation of cationic dyes in dye wastewater. The adsorption performance and mechanism of MT on methylene blue (MB) and crystal violet (CV) were investigated by batch adsorption experiments. The results demonstrated that the highest adsorption values of MT for MB (411 mg/g) and CV (553 mg/g) were greatly higher than the reported values of other biosorbents. In addition, the adsorption behaviors of methylene blue (MB) and crystal violet (CV) by MT were spontaneous exothermic reactions and closely followed the pseudo-second-order (PSO) kinetics and Langmuir isotherm. Further, the depleted MT was regenerated using pyrolysis mode to convert depleted MT into MT-biochar (MBC). The maximum adsorption of Cu2+ and Pb2+ by MBC was up to 320 mg/g and 840 mg/g, respectively. In conclusion, this work presented a new option for the adsorption of cationic dyes in wastewater and a new perspective for the treatment of depleted biosorbents.

Vacancy-enhanced oxygen redox and structural stability of spinel Li2Mn3O7-x.

Vacancies have been proved effective in activating the oxygen redox and stabilizing the structure of the oxide cathode materials for the Na-ion batteries, but their effect on the cathode materials of the Li-ion batteries is unclear. We herein show that they have similar effect on spinel [Li4/7Mn2/7□1/7]8a[Li4/7Mn10/7]16d[O4-x']32e.

Feasibility to Improve the Stability of Lithium-Rich Layered Oxides by Surface Doping.

Li-rich layer-structured oxides are considered promising cathode materials for their specific capacities above 250 mAh·g-1. However, the drawbacks such as poor rate performance, fast capacity fading, and the continuous transition metal (TM) migration into the Li layer hinder their commercial applications. To address these issues, surface doping of Ti and Zr was conducted to the Li- and Mn-rich layered oxide (LMR), Li1.2Mn0.54Ni0.13Co0.13O2. The drop of the average discharge potentials of the Ti- and Zr-doped LMR was reduced by 593 and 346 mV in 100 cycles, respectively. With aberration-corrected scanning transmission electron microscopy and electron energy loss spectroscopy, we clarified that Ti4+ and Zr4+ ions are located near the surface of the material, anchor the surface oxygen, and stabilize the LMR structure. The difference in the strengths of the Ti-O and Zr-O bonds and the doping-resultant electronic structures were determined with density functional theory (DFT) calculations and soft X-ray absorption spectroscopy (SXAS), responsible for the electrochemical performance of surface-doped materials. These findings verify our modification strategies to enhance the cycling performances of the promising LMR cathode materials.

Real-Time Remaining Useful Life Prediction of Cutting Tools Using Sparse Augmented Lagrangian Analysis and Gaussian Process Regression.

Remaining useful life (RUL) of cutting tools is concerned with cutting tool operational status prediction and damage prognosis. Most RUL prediction methods utilized different features collected from different sensors to predict the life of the tool. To increase the prediction accuracy, it is often necessary to mount a great deal of sensors on the machine in order to collect more types of signals, which can heavily increase the cost in industrial applications. To deal with this issue, this study, for the first time, proposed a new feature network dictionary, which can enlarge the number of candidate features under limited sensor conditions, and the developed dictionary can potentially contain as much useful information as possible. This process can replace the installation of more sensors and incorporate more information. Then, the sparse augmented Lagrangian (SAL) feature selection method is proposed to reduce the number of candidate features and select the most significant features. Finally, the selected features are input to the Gaussian Process Regression (GPR) model for the RUL estimation. Extensive experiments demonstrate that our proposed RUL estimation framework output performs traditional methods, especially for the cost savings for on-line RUL estimation.

Thin-Film Composite Membrane Prepared by Interfacial Polymerization on the Integrated ZIF-L Nanosheets Interface for Pervaporation Dehydration.

Thin-film composite (TFC) membranes are attracting wide attention because their ultrathin selective layer usually corresponds to the higher membrane flux for pervaporation. However, the direct preparation of the TFC membranes on ceramic substrates confronted with the great difficulties because the larger pores on ceramic substrate surfaces are detrimental to the formation of an intact polyamide (PA) selective layer produced by interfacial polymerization (IP) reaction. Here, the integrated ZIF-L nanosheets were proposed to be used as an assistance interlayer for the first time to eliminate the existence of the pores of the ceramic support, and provides a better basis for the formation of an intact PA selective layer by IP reaction between TMC and ethylenediamine (EDA). The experimental data obtained in pervaporation (PV) show that the increased flux from 1.1 to 2.9 kg/m2h corresponds to the decreased separation factor from 396 to 110 when the feed concentration of ethanol decreases from 95 wt % to 80 wt % at 50 °C. In addition, the membrane flux increases from 0.8 to 2.5 kg/m2h with a change of the separation factor from 683 to 111 when the operational temperature varies from 30 to 60 °C. These results demonstrate the great potential of the fabricated TFC membranes in practical application for PV dehydration of organic solutions.

Competitive Solvation Enhanced Stability of Lithium Metal Anode in Dual-Salt Electrolyte.

The development of lithium metal batteries is hindered by the low Coulombic efficiency and poor cycling stability of the metallic lithium. The introduction of consumptive LiNO3 as an additive can improve the cycling stability, but its low solubility in the carbonate electrolytes makes this strategy impractical for long-term cycling. Herein we propose LiNO3 as a cosalt in the LiPF6-LiNO3 dual-salt electrolyte to enhance the cycling stability of lithium plating/stripping. Competitions among the components and the resultant substitution of NO3- for PF6- in the solvation shell facilitate the formation of a Li3N-rich solid electrolyte interphase (SEI) film and suppress the LiPF6 decomposition. The highly Li+ conductive and stable SEI film effectively tailors the lithium nucleation, suppresses the formation of lithium dendrites, and improves the cycling performance. The competitive solvation has profound importance for the design of a complex electrolyte to meet the multiple requirements of secondary lithium batteries.

Metal-free catalytic hydrocarboxylation of hexafluorobut-2-yne.

An efficient method for stereoselective synthesis of trifluorinated enol esters catalyzed by base was introduced. The DFT calculations and experimental results both supported the nucleophilic addition process. The protocol featured mild reaction conditions and showed a wide functional group tolerance. The one-pot simultaneous etherification and esterification of the salicylic acids further demonstrated the prospective synthetic application.

Functional differences of three CXCL10 homologues in the giant spiny frog Quasipaa spinosa.

Chemokines are a superfamily of structurally related chemotactic cytokines exerting significant roles in acting as a bridge between the innate and adaptive immune responses. In this study, we identified three CXC motif chemokine 10 (CXCL10) homologues (QsCXCL10-1, QsCXCL10-2 and QsCXCL10-3) from giant spiny frog Quasipaa spinosa. All three deduced QsCXCL10 proteins contained four conserved cysteine residues as found in other known CXC chemokines. Phylogenetic analysis showed that QsCXCL10-1, 2, 3 and other CXCL10s in amphibian were grouped together to form a separate clade. These three QsCXCL10s were highly expressed in spleen and blood. Upon infection with Staphylococcus aureus or Aeromonas hydrophila, the expressions of QsCXCL10s were markedly increased in spleen and blood during biotic stresses. Meanwhile, the QsCXCL10s transcription in liver could also be up-regulated under abiotic stresses such as cold and heat stresses. The recombinant proteins of frog CXCL10 homologues were produced and purified in E. coli and possessed similar but differential bioactivities. Both rCXCL10-1 and rCXCL10-2 had strong effects on the up-regulation of pro-inflammatory cytokines (TNF-α, IL-1ß and IL-8) in vivo, whereas rCXCL10-3 induced a weak expression of these cytokines. Moreover, the rCXCL10-1 and rCXCL10-2 could strongly promote splenocyte proliferation and induce lymphocytes migration, while rCXCL10-3 had limited effects on these biological processes. All three frog chemokines triggered their functional activities by engaging CXC motif chemokine receptor 3 (CXCR3). Taken together, these results revealed that the three QsCXCL10s had similar but differential functional activities in mediating immune responses and host defenses, which might contribute to a better understanding of the functional evolution of CXCL10 in vertebrates.

The first evidence of four transcripts from two Interleukin 18 genes in animal and their involvement in immune responses in the largest amphibian Andrias davidianus.

Interleukin 18 (IL-18), a member of IL-1 cytokine superfamily, is an important proinflammatory cytokine with multiple functions in both innate immunity and acquired immunity. However, the characteristics and functional roles of IL-18 remain largely unknown in amphibians, which were classed as major group of vertebrates. In the present study, two IL-18 genes (AdIL-18A and AdIL-18B) and four transcripts (AdIL-18A1, AdIL-18A2, AdIL-18B1 and AdIL-18B2) were firstly identified and characterized from Chinese giant salamander (Andrias davidianus). To the best of our knowledge, this is the first report on the presence of more than one gene copy or two transcripts of IL-18 in one species. The complete open reading frames of AdIL-18A1, AdIL-18A2, AdIL-18B1 and AdIL-18B2 were 588 bp, 603 bp, 591 bp and 606 bp, respectively. The putative AdIL-18 proteins possessed the typical IL-1 domains and phylogenetic analysis indicated that AdIL-18s grouped together with other vertebrate IL-18 proteins. The expression profiles of AdIL-18s were investigated under the challenges of Aeromonas hydrophila, Staphylococcus ureae and Poly (I:C) respectively, and the results suggested that AdIL-18s were involved in the immune responses against both bacterial and viral infections. Moreover, the expression levels of two NF-κBs (P100 and P105) and four proinflammatory cytokines (IL-1ß, IL-6, TNF-α and IFN-γ) were inhibited in AdIL-18A1/A2-silenced cells when treated with bacteria and viral RNA analog. Additionally, the transcription levels of these immune-related cytokine genes were markedly induced when the lymphocytes were treated with recombinant AdIL-18A1 or AdIL-18A2 proteins, implying the involvement of AdIL-18s in triggering NF-κB signaling and proinflammatory responses. These results might provide new insights into the origin or evolution of IL-18 in amphibians and even in vertebrates.

[Research advance in circular RNAs].

Circular RNAs (circRNAs) are a class of endogenous, covalently closed, single-stranded RNA without 3'-poly(A) and 5'-cap structures. CircRNAs are characterized by universality, diversity, stability and conservation, and have been found to regulate mammalian transcription and be translated into proteins. In this review, we summarized the biogenesis, classification, expression, distribution, biological functions and regulation of circRNAs. In addition, we discussed the association of circRNAs with diseases and the methods for identification and characterization of circRNAs. Finally, we speculated the application prospect and research direction of circRNAs.

The nuclear factor interleukin 3-regulated (NFIL3) transcription factor involved in innate immunity by activating NF-κB pathway in mud crab Scylla paramamosain.

NFIL3 is a transcriptional activator of the IL-3 promoter in T cells. In vertebrates, it has been characterized as an essential regulator of several cellular processes such as immunity response, apoptosis and NK cells maturation. However, the identification and functional characterization of NFIL3 still remains unclear in arthropods. In this study, the NFIL3 homologue was firstly cloned and characterized in mud crab Scylla paramamosain. The full-length of SpNFIL3 was 2, 041 bp in length with an open reading frame of 1, 509 bp, containing a conserved basic region of leucin zipper domain. The qRT-PCR analysis indicated that SpNFIL3 was significantly highly expressed in hepatopancreas and in hemocytes. Moreover, the SpNFIL3 transcription could be up-regulated after the challenge of Vibrio alginolyticus or virus-analog Poly (I:C). The dual-luciferase reporter assays revealed that SpNFIL3 could activate NF-κB pathway. The immunofluorescence assay indicated SpNFIL3 was located in nucleus. After NFIL3 was interfered in vivo and in vitro, the expressions of two NF-κB members (SpRelish and SpDorsal), six antimicrobial peptide genes (SpCrustin and SpALF2-6) and pro-inflammatory cytokine SpIL-16 were suppressed, and the bacteria clearance capacity of crabs was also markedly impaired in NFIL3 silenced crabs. These results indicated that SpNFIL3 played crucial role in the innate immunity of S. paramamosain and it also brought new insight into the origin and evolution of NFIL3 in arthropods and even in invertebrates.

The Activin-like ligand Dawdle regulates innate immune responses through modulating NF-κB signaling in mud crab Scylla paramamosain.

Activins, members of transforming growth factor ß (TGF-ß) superfamily, are pleiotropic cytokines with critical roles in mediating cell proliferation, differentiation, homeostasis, apoptosis and immune response. However, the structural characteristics and specific functions of Activins remain largely unknown in invertebrates. In the present study, an Activin-like ligand Dawdle (Daw) was firstly identified and characterized from mud crab Scylla paramamosain. The obtained cDNA sequence of SpDaw was 2, 196 bp long with a 1, 149 bp open reading fame, which encoded a putative protein of 382 amino acids. The putative SpDaw protein contained a signal peptide, a TGF-ß propeptide region and a TGF-ß domain. Real-time PCR analysis demonstrated that SpDaw was predominantly expressed at early embryonic development stage and premolt stages, implying its participation in development and growth. Furthermore, SpDaw responded to both Vibro alginolyticus and Poly (I:C) challenges, suggesting the involvement of SpDaw in innate immune responses. Knockdown of SpDaw in vivo dramatically increased the expressions of NF-κB signaling genes and anti-lipopolysaccharide factor (ALF) genes, and the bacteria clearance efficiency was also markedly enhanced in SpDaw-silenced crabs. Moreover, the in vitro experiment further demonstrated that recombinant SpDaw protein could block the increased transcription of IKKs, NF-κBs and ALFs induced by pathogen challenges. Taken together, these results indicated that SpDaw not only participated in development and growth processes but also played an immune-regulatory role in crabs' innate immunity, which may pave the way for a better understanding of TGF-ß superfamily members in crustacean species.

Transforming growth factor-ß-activating kinase 1 and its binding protein 1 participate in the innate immune responses via modulating the IMDNFκB signaling in mud crab (Scylla paramamosain).

Transforming growth factor-ß-activating kinase 1 (TAK1) is essential for diverse important biological functions, such as innate immunity, development and cell survival. In the present study, the homologs of TAK1 and TAK1-binding protein 1 (TAB1) were identified and characterized from mud crab Scylla paramamosain for the first time. The full-length cDNAs of SpTAK1 and SpTAB1 were 2, 226 bp and 2, 433 bp with 1, 782 bp and 1, 533 bp open reading frame (ORF), respectively. The deduced SpTAK1 protein contained a conserved S_TKc (Serine/threonine protein kinases, catalytic) domain, and the putative SpTAB1 protein possessed a typical PP2Cc (Serine/threonine phosphatases, family 2C, catalytic) domain and a potential TAK1 docking motif. Real-time PCR analysis showed that SpTAK1 and SpTAB1 were highly expressed at early development stages, suggesting their participation in crab's development process. Moreover, the expression levels of SpTAK1 and SpTAB1 in hepatopancreas were positively stimulated after challenge with Vibro alginolyticus and Poly (I:C), implying the involvement of SpTAK1 and SpTAB1 in innate immune responses against both bacterial and viral infections. When SpTAK1 or SpTAB1 were silenced in vivo, the expression levels of two IMDNFκB signaling components (SpIKKß and SpRelish) and six antimicrobial peptide (AMP) genes (SpALF1-5 and SpCrustin) were significantly reduced, and the bacteria clearance capacity of crabs was also markedly impaired in SpTAK1 or SpTAB1 silenced crabs. Additionally, overexpression of SpTAK1 and SpTAB1 in HEK293T cells could markedly activate the mammalian NF-κB signaling. Collectively, our results suggested that TAK1 and TAB1 regulated crab's innate immunity via modulating the IMDNFκB signaling. These findings may provide new insights into the TAK1/TAB1-mediated signaling cascades in crustaceans and pave the way for a better understanding of crustacean innate immune system.

Effects of both cold and heat stress on the liver of the giant spiny frog (Quasipaa spinosa): stress response and histological changes.

Ambient temperature-associated stress can affect normal physiological functions in ectotherms. To assess the effects of cold or heat stress on amphibians, giant spiny frogs (Quasipaa spinosa) were acclimated at 22°C followed by exposure to 5°C or 30°C for 0, 3, 6, 12, 24 and 48 h, respectively. Histological alterations, apoptotic index, generation of mitochondrial reactive oxygen species (ROS), antioxidant activity indices and stress-response gene expression in frog livers were subsequently determined. Results showed that many fat droplets appeared after 12 h of heat stress and the percentage of melanomacrophage centres significantly changed after 48 h at both stress conditions. Furthermore, the mitochondrial ROS levels were elevated in a time-dependent manner up to 6 h and 12 h in the cold and heat stress groups, respectively. The activities of superoxide dismutase, glutathione peroxidase and catalase were successively increased with increasing periods of cold or heat exposure, and their gene expression levels showed similar changes in both stress conditions. Most tested heat shock protein (HSP) genes were sensitive to temperature exposure, and the expression profiles of most apoptosis-related genes was significantly upregulated at 3 and 48 h under cold and heat stress, respectively. Apoptotic index at 48 h under cold stress was significantly higher than that under heat stress. Notably, lipid droplets, HSP30, HSP70 and HSP110 might be suitable bioindicators of heat stress. The results of these alterations at physiological, biochemical and molecular levels might contribute to a better understanding of the stress response of Q. spinosa, and perhaps amphibians more generally, under thermal stress.

Effects of heat stress on the liver of the Chinese giant salamander Andrias davidianus: Histopathological changes and expression characterization of Nrf2-mediated antioxidant pathway genes.

Nuclear factor E2-related factor 2 (Nrf2) is a crucial transcription factor that regulates the basal and inducible expression of many antioxidant-relevant genes, and the Nrf2-mediated antioxidant pathway has been regarded as a critical switch in the initiation of cellular defence systems against oxidative damages. In this study, Nrf2 was first identified and characterized in the Chinese giant salamander (Andrias davidianus). A. davidianus was exposed to a high ambient temperature of 30 °C for various periods of time (0, 3, 6, 12, 24, 48 and 72 h). We investigated the effects of heat stress on alterations of the hepatic malondialdehyde (MDA) concentration, the activities of lactic acid dehydrogenase (LDH), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD), the histology of the liver, and the mRNA expression patterns of 11 genes involved in the Nrf2-mediated antioxidant pathway in A. davidianus. The results showed that both the hepatic LDH activity and MDA content significantly increased after heat exposure, indicating that heat stress could induce cell injury and oxidative damage. Histological analysis of the liver showed that heat stress caused hepatocyte abnormalities, fat accumulation and ultrastructural alterations of the hepatocytes, endoplasmic reticulum and nuclei. The expression patterns of genes involved in the Nrf2-mediated antioxidant pathway in the liver were distinct when A. davidianus was exposed to heat stress. To the best of our knowledge, this study is the first on the characterization of Nrf2 in A. davidianus and even in amphibians. The results indicated that heat stress could induce oxidative damage, and the Nrf2 antioxidant pathway might play a critical role in the resistance against heat stress in A. davidianus. These findings will deepen and enrich the current knowledge on the evolutionary conserved antioxidant roles and mechanisms of Nrf2 in A. davidianus, or even in amphibians, in the antioxidant defence against heat stress.