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Localized surface plasmon resonances (LSPRs) can enhance the electromagnetic fields on metallic nanostructures upon light illumination, providing an approach for manipulating light-matter interactions at the sub-wavelength scale. However, currently, there is no thorough investigation of the physical mechanism in the dynamic formation of the strongly coupled LSPRs on sub-5 nm plasmonic cavities at the sub-picosecond scale. In this work, through femtosecond broadband transient absorption spectroscopy, we reveal the dynamic ultrastrong coupling processes in a nanoparticle-in-trench (NPiT) structure containing 2 nm gap cavities, and demonstrate a coherent motional coupling between vibrating AuNPs and the nanogaps. We achieve a maximum Rabi splitting energy of â¼660 meV in the sub-picosecond hot-electron relaxation time scale under the resonant excitation of the nanogap cavity's LSPR, reaching the ultrastrong coupling regime. This leads to a change of global vibration modes for the 2 nm gap cavity, potentially related to the dynamical Casimir effect with nanogap resonators.
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Multimode fiber lasers have become a new platform for investigating nonlinear phenomena since the report on spatiotemporal mode-locking. In this work, the multimode soliton pulsation with a tunable period is achieved in a spatiotemporal mode-locked fiber laser. It demonstrates that the pulsation period drops while increasing the pump power. Moreover, it is found that different transverse modes have the same pulsation period, asynchronous pulsation evolution and different dynamical characteristics through the spatial sampling technique and the dispersive Fourier transform technique. To further verify the experimental results, we numerically investigate the influences of the gain and the loss on the pulsation properties. It is found that within a certain parameter range, the pulsation period drops and rises linearly with the increase of the gain and the loss, respectively. The obtained results contribute to understanding the formation and regulating of soliton pulsations in fiber lasers.
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Spatiotemporal mode-locked (STML) fiber lasers have become a new platform for investigating nonlinear phenomena. In this work, spatiotemporal dual-periodic soliton pulsation (SDSP) is firstly observed in an STML fiber laser. It is found that in the SDSP, the long-period pulsations (LPPs) of different transverse modes are synchronous, while the short-period pulsations (SPPs) exhibit asynchronous modulations. The numerical simulation confirms the experimental results and further reveals that the proportion of transverse mode components can manipulate the periods of the LPP and SPP but does not affect the synchronous and asynchronous pulsations of different transverse modes. The obtained results bring the study of spatiotemporal dissipative soliton pulsation into the multi-period modulation stage, which helps to understand the complex spatiotemporal dynamics in STML fiber lasers and discover new dynamics in high-dimensional nonlinear systems.
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Laminin receptor (LR), which mediating cell adhesion to the extracellular matrix, plays a crucial role in cell signaling and regulatory functions. In the present study, a laminin receptor gene (SpLR) was cloned and characterized from the mud crab (Scylla paramamosain). The full length of SpLR contained an open reading frame (ORF) of 960 bp encoding 319 amino acids, a 5' untranslated region (UTR) of 66 bp and a 3' UTR of 49 bp. The predicted protein comprised two Ribosomal-S2 domains and a 40S-SA-C domain. The mRNA of SpLR was highly expressed in the gill, followed by the hepatopancreas. The expression of SpLR was up-regulated after mud crab dicistrovirus-1(MCDV-1) infection. Knocking down SpLR in vivo by RNA interference significantly down-regulated the expression of the immune genes SpJAK, SpSTAT, SpToll1, SpALF1 and SpALF5. This study shown that the expression level of SpToll1 and SpCAM in SpLR-interfered group significantly increased after MCDV-1 infection. Moreover, silencing of SpLR in vivo decreased the MCDV-1 replication and increased the survival rate of mud crabs after MCDV-1 infection. These findings collectively suggest a pivotal role for SpLR in the mud crab's response to MCDV-1 infection. By influencing the expression of critical innate immune factors and impacting viral replication dynamics, SpLR emerges as a key player in the intricate host-pathogen interaction, providing valuable insights into the molecular mechanisms underlying MCDV-1 pathogenesis in mud crabs.
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Sequência de Aminoácidos , Proteínas de Artrópodes , Braquiúros , Regulação da Expressão Gênica , Imunidade Inata , Filogenia , Receptores de Laminina , Alinhamento de Sequência , Animais , Braquiúros/genética , Braquiúros/imunologia , Receptores de Laminina/genética , Receptores de Laminina/imunologia , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Proteínas de Artrópodes/química , Imunidade Inata/genética , Regulação da Expressão Gênica/imunologia , Alinhamento de Sequência/veterinária , Perfilação da Expressão Gênica/veterinária , Sequência de BasesRESUMO
We report, for the first time to the best of our knowledge, a spatiotemporal mode-locked (STML) multimode fiber laser based on nonlinear amplifying loop mirror (NALM), generating dissipative soliton resonance (DSR) pulses. Due to the complex filtering characteristics caused by the inherent multimode interference filtering structure and NALM in the cavity, the STML DSR pulse has wavelength tunable function. What's more, kinds of DSR pulses are also achieved, including multiple DSR pulses, and the period doubling bifurcations of single DSR pulse and multiple DSR pulses. These results contribute to further understand the nonlinear properties of STML lasers and may shed some light on improving the performance of the multimode fiber lasers.
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Cytochrome P450 (CYPs) enzymes are one of the critical detoxification enzymes, playing a key role in antioxidant defense. However, the information of CYPs cDNA sequences and their functions are lacked in crustaceans. In this study, a novel full-length of CYP2 from the mud crab (designated as Sp-CYP2) was cloned and characterized. The coding sequence of Sp-CYP2 was 1479 bp in length and encoded a protein containing 492 amino acids. The amino acid sequence of Sp-CYP2 comprised a conserved heme binding site and chemical substrate binding site. Quantitative real-time PCR analysis revealed that Sp-CYP2 was ubiquitously expressed in various tissues, and it was highest in the heart followed by the hepatopancreas. Subcellular localization showed that Sp-CYP2 was prominently located in the cytoplasm and nucleus. The expression of Sp-CYP2 was induced by Vibrio parahaemolyticus infection and ammonia exposure. During ammonia exposure, ammonia exposure can induce oxidative stress and cause severely tissue damage. Knocking down Sp-CYP2 in vivo can increase malondialdehyde content and the mortality of mud crabs after ammonia exposure. All these results suggested that Sp-CYP2 played a crucial role in the defense against environmental stress and pathogen infection in crustaceans.
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Braquiúros , Animais , Antioxidantes , Sequência de Bases , Filogenia , Amônia , Imunidade Inata/genética , Proteínas de ArtrópodesRESUMO
Heat shock proteins play an important role in host defense, and modulate immune responses against pathogen infection. In this study, a novel HSC70 from the mud crab (designated as SpHSC70) was cloned and characterized. The full length of SpHSC70 contained a 58 bp 5'untranslated region (UTR), an open reading frame (ORF) of 2,046 bp and a 3'UTR of 341 bp. The SpHSC70 protein included the conserved DnaK motif. The mRNA of SpHSC70 was highly expressed in the hemocytes, heart and hepatopancreas, and lowly expressed in the intestine. The subcellular localization results indicated that SpHSC70 was localized in both the cytoplasm and the nucleus. Moreover, SpHSC70 was significantly responsive to bacterial challenge. RNA interference experiment was designed to investigate the roles of SpHSC70 in response to bacterial challenge. V. parahaemolyticus infection induced the expression levels of SpPO, SpHSP70, SpSOD and SpCAT. Knocking down SpHSC70 in vivo can decrease the expression of these genes after V. parahaemolyticus infection. These results suggested that SpHSC70 could play a vital role in defense against V. parahaemolyticus infection via activating the immune response and antioxidant defense signaling pathways in the mud crab.
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Braquiúros , Vibrioses , Vibrio parahaemolyticus , Animais , Vibrio parahaemolyticus/fisiologia , Vibrioses/microbiologia , Interferência de RNA , Bactérias/metabolismo , Proteínas de Artrópodes , FilogeniaRESUMO
Activating transcription factor 6 (ATF6) is critical for regulation of unfolded protein response (UPR), which is involved in the endoplasmic reticulum (ER) proteostasis maintenance and cellular redox regulation. In the present study, a ATF6 gene from the mud crab (designated as Sp-ATF6) has been cloned and identified. The open reading frame of Sp-ATF6 was 1917 bp, encoding a protein of 638 amino acids. The deduced amino acid sequences of Sp-ATF6 contained a typical basic leucine zipper (BZIP domain). Sp-ATF6 was widely expressed in all tested tissues, with the highest expression levels in the hemocytes and the lowest in the muscle. Subcellular localization showed that Sp-ATF6 was expressed in both nucleus and cytoplasm of S2 cells. The expression level of Sp-ATF6 was induced by hydrogen peroxide and V. parahaemolyticus challenge, indicating that the ATF6 pathway was activated in response to ER stress. In order to know more about the regulation mechanism of the Sp-ATF6, RNA interference experiment was investigated. Knocking down Sp-ATF6 in vivo can decrease the expression of antioxidant-related genes (CAT and SOD) and heat shock proteins (HSP90 and HSP70) after V. parahaemolyticus infection. All these results suggested that Sp-ATF6 played a crucial role in the defense against environmental stress and pathogen infection in crustaceans.
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Braquiúros , Animais , Braquiúros/microbiologia , Peróxido de Hidrogênio , Fator 6 Ativador da Transcrição/genética , Fator 6 Ativador da Transcrição/metabolismo , Filogenia , Sequência de Aminoácidos , Bactérias/metabolismo , Proteínas de Artrópodes/química , Imunidade Inata/genéticaRESUMO
We report a narrow bandwidth spatiotemporal mode-locked (STML) ytterbium-doped fiber laser, based on a homemade carbon nanotube/polyvinyl alcohol composite film and the multimode interference filtering effect. The wavelength-tunable narrow bandwidth STML operations combined with different pulse states are achieved, including single pulse, multiple pulses, and harmonics. The 3-dB bandwidth at the single-pulse state is 103 pm, while at the harmonic state, it is as narrow as 26 pm. To give an insight into the generation of the narrow bandwidth STML pulses, numerical simulations are performed. Such a laser has a wide range of potential applications in fields of optical communication and optical measurement, as well as provides a favorable platform for studying the evolution dynamics of multimode solitons.
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Glutaredoxin (Grx) is a class molecule oxidoreductase, which plays a key role in maintaining redox homeostasis and regulating cell survival pathways. However, the expression pattern and function of Grx remain unknown in the mud crab (Scylla paramamosain). In the present study, a novel full-length of Grx 5 from the mud crab (designated as Sp-Grx 5) was cloned and characterized. The open reading frame of Sp-Grx 5 was 441 bp, which encoded a putative protein of 146 amino acids. The amino acid sequence of Sp-Grx 5 contained a typical C-G-F-S redox active motif and several GSH binding sites. Sp-Grx 5 widely existed in all tested tissues with a high-level expression in hepatopancreas. Subcellular localization showed that Sp-Grx 5 was located in the cytoplasm and nucleus. The expression of Sp-Grx 5 was significantly up-regulated after Vibrio parahaemolyticus infection and cadmium exposure, suggesting that Sp-Grx 5 was involved in innate immunity and detoxification. Furthermore, overexpression of Sp-Grx 5 could improve cells viability after H2O2 exposure. All these results indicated that Sp-Grx 5 played important roles in the redox homeostasis and innate immune response in crustaceans.
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Braquiúros , Aminoácidos , Animais , Proteínas de Artrópodes/química , Bactérias/metabolismo , Sequência de Bases , Cádmio/toxicidade , Glutarredoxinas/genética , Peróxido de Hidrogênio , Imunidade Inata/genética , FilogeniaRESUMO
Phosphofructokinase (PFK), the key enzyme of glycolysis, can catalyze the irreversible transphosphorylation of fructose-6-phosphate forming fructose-1, 6-biphosphate. In the present study, a PFK gene from the mud crab Scylla paramamosain, named SpPFK, was cloned and characterized. The full length of SpPFK contained a 5'untranslated region (UTR) of 249 bp, an open reading frame of 2,859 bp, and a 3'UTR of 1,248 bp. The mRNA of SpPFK was highly expressed in the gill, followed by the hemocytes and muscle. The expression of SpPFK was significantly up-regulated after mud crab dicistrovirus-1 (MCDV-1) infection. Knocking down SpPFK in vivo by RNA interference significantly reduced the expression of lactate dehydrogenase after MCDV-1 infection. Furthermore, silencing of SpPFK in vivo increased the survival rate of mud crabs and decreased the MCDV-1 copies in the gill and hepatopancreas after MCDV-1 infection. All these results suggested that SpPFK could play an important role in the process of MCDV-1 proliferation in mud crab.
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Braquiúros , Animais , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/metabolismo , Braquiúros/genética , Braquiúros/metabolismo , Proliferação de Células , Fosfofrutoquinases/genética , Fosfofrutoquinases/metabolismo , FilogeniaRESUMO
Oxidative stress is considered as the toxicity mechanism of environmental stressors on aquatic organisms. This study aims to explore the effects of oxidative stress on physiological responses, DNA damage and transcriptional profiles of the mud crabs Scylla paramamosain. In the present study, mud crabs were injected with 0.1% and 1% hydrogen peroxide (H2O2) for 72 h. The results showed that superoxide dismutase and catalase activities significantly decreased after H2O2 injection. Malondialdehyde content, H2O2 content, aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase activity significantly increased after H2O2 injection. Moreover, DNA damage occurred after H2O2 injection. Transcriptome analysis showed that 531 and 372 differentially expressed genes (DEGs) were identified after 0.1% and 1% H2O2 injection, respectively. These DEGs were mainly involved in the oxidative stress response and immune functions. All these results indicated that oxidative stress could impair both antioxidant defense systems and immune systems. Transcriptome analysis provided valuable information on gene functions associated with the response to oxidative stress in the mud crab.
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Braquiúros , Dano ao DNA/efeitos dos fármacos , Peróxido de Hidrogênio/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Animais , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , TranscriptomaRESUMO
Apoptosis plays essential roles in the immune defense mechanism against pathogen infection. Caspase 3 is a family of cysteine proteases involved in apoptosis and the immune response. In this study, the full-length of mud crab (Scylla paramamosain) caspase 3 (designated as Sp-caspase 3) was cloned and characterized. The open reading frame of Sp-caspase 3 was comprised a 1035 bp, which encoded a putative protein of 344 amino acids. Sp-caspase 3 was ubiquitously expressed in various tissues with a high-level expression in hemocytes. Cellular localization analysis revealed that Sp-caspase 3 was located in the cytoplasm and nucleus. Over-expression of Sp-caspase 3 could induce cell apoptosis. In addition, V. Parahaemolyticus infection induced the relative expression of caspase-3 mRNA and increased caspase-3 activity. Knocking down Sp-caspase 3 in vivo significantly reduced cell apoptosis and increased mortality of mud crab after V. parahaemolyticus infection. These results indicated that Sp-caspase 3 played important roles in the immune response and apoptosis against bacterial infection.
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Braquiúros , Caspase 3 , Vibrioses , Vibrio parahaemolyticus , Animais , Proteínas de Artrópodes/metabolismo , Braquiúros/enzimologia , Braquiúros/imunologia , Braquiúros/microbiologia , Caspase 3/metabolismo , Filogenia , Vibrioses/imunologia , Vibrioses/veterinária , Vibrio parahaemolyticus/imunologiaRESUMO
Circular RNA (circRNA) molecules contain microRNA (miRNA) response elements that are able to competitively bind miRNAs as well as function as miRNA sponges within cells, which can reduce miRNA inhibition of target genes, thereby increasing their expression. TargetScan and miRanda bioinformatic tools were used to analyze the binding sites between genes. The relative levels of gene expression in tissues and cells were verified using quantitative reverse transcription-polymerase chain reaction. Inhibition of cell proliferation was detected using a WST-8 method. Cell invasion ability and migration ability were assessed using a Transwell migration assay and a scratch assay, respectively. The binding of miRNA and circRNA was detected using an RNA pull-down assay. Bifluorescence reporter gene vectors were constructed to verify the binding of miRNA to messenger RNA. A tumor model of cervical cancer cell transplantation in mice was constructed to observe the effect of the genes on tumor growth. hsa_circ_0031288 and B-cell CLL/lymphoma 6 (Bcl-6) exhibited high expression in cervical cancer cells and tissue, while hsa-miR-139-3p exhibited low expression. Reducing hsa_circ_0031288 and Bcl-6 expression or increasing hsa-miR-139-3p expression significantly inhibited the migration, invasion, proliferation, and growth of xenograft and HeLa cells. hsa_circ_0031288 had a regulatory effect on hsa-miR-139-3p, and hsa-miR-139-3p targeted the 3' untranslated region of Bcl-6. Reducing hsa_circ_0031288 expression promoted hsa-miR-139-3p expression, while overexpressing miR-139-3p inhibited the transcription of Bcl-6. In the cervical cancer HeLa cell line, the hsa_circ_0031288/hsa-miR-139-3p/Bcl-6 regulatory axis affects cell migration and proliferation, and its mechanism may involve hsa_circ_0031288 acting as a sponge for hsa-miR-139-3p, thereby relieving the transcriptional inhibition of Bcl-6. This suggests an approach for elucidating the pathogenesis of cervical cancer while offering new intervention targets for cervical cancer treatment.
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Movimento Celular/genética , Retroalimentação Fisiológica , MicroRNAs/metabolismo , Proteínas Proto-Oncogênicas c-bcl-6/metabolismo , RNA Circular/metabolismo , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Animais , Feminino , Regulação Neoplásica da Expressão Gênica , Células HEK293 , Células HeLa , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , Invasividade Neoplásica/genética , Proteínas Proto-Oncogênicas c-bcl-6/genética , Interferência de RNA , RNA Circular/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/genética , Carga Tumoral/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
This research is to predict anti-Alzheimer's disease active constituents on the target of acetylcholinesterase(AChE) from Glycyrrhizae Radix et Rhizoma with the help of pharmacophore and molecular docking. AChE ligand-based pharmacophore model was set up and the molecular library of the constituents from Glycyrrhizae Radix et Rhizoma were established by collecting literature. Then the constituents from Glycyrrhizae Radix et Rhizoma were screen for the potential AChE inhibitory potency in silico through matching with the best pharmacophore model. The flexible docking was used to evaluate the interactions between compounds screened from pharmacophore model and AChE protein(PDB ID:4 EY7). The interactions were expressed including but not limited to CDOCKER interaction energy, hydrogen bonds and non-bonding interactions. The molecular library of Glycyrrhizae Radix et Rhizoma contains 44 chemical constituents. As for the pharmacophore model, six kinds of potential AChE inhibitory constituents from Glycyrrhizae Radix et Rhizoma were considered to be the promising compounds according to the results of searching 3 D database of pharmacophore model. The molecular docking was possessed and the interaction patterns were given to show the detail interactions. The compounds screening from the pharmacophore model were consistent with the existing studies to some degree, indicating that the virtual screen protocols of AChE inhibitory constituents from Glycyrrhizae Radix et Rhizoma based on pharmacophore and molecular docking was reliable.
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Medicamentos de Ervas Chinesas , Glycyrrhiza , Triterpenos , Simulação de Acoplamento Molecular , RizomaRESUMO
Malaria information was collected through online systems in Baoying County from 2005 to 2014 and descriptive analysis was performed on the data. A total of 175 cases of malaria were reported in the years, with no report of death. During 2005-2008, vivax malaria cases constituted 86.4%ï¼19/22ï¼ of all cases reported, and all infected locally with diease onset mainly occurred from May to November. During 2009-2014, falciparum malaria cases constituted 82.4%ï¼126/153ï¼, and all imported cases, which showed a trend of increase, from one case in 2007 to 22 in 2009. All the cases reported during 2010-2014 were imported cases. As to malaria control, there was an overall regular treatment rate of 100%, 195 patients had radical cure during the resting stage, and blood test was conducted in 52 827 fevered patients, resulting in a positive rate of 0.3%ï¼149/52 827ï¼. Although no local malaria cases were reported since 2010, the imported malaria is still an important issue.
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Malária/epidemiologia , China , HumanosRESUMO
Hypoxia-inducible factors -1 (HIF-1) is a crucial transcription factor that regulates the expression of glycolytic genes. Our previous study proved that the Mud crab dicistrovirus-1 (MCDV-1) can induce aerobic glycolysis that favors viral replication in mud crab Scylla paramamosain. However, the role of HIF-1 on key glycolytic genes during the MCDV-1 infection has not been examined. In this study, the intricate interplay between HIF-1 and the key glycolysis enzyme, lactate dehydrogenase (LDH), was investigated after MCDV-1 infection. The expression of LDH was significant increased after MCDV-1 infection. Additionally, the expression of HIF-1α was upregulated following MCDV-1 infection, potentially attributed to the downregulation of prolyl hydroxylase domains 2 expression. Subsequent examination of the SpLDH promoter identified the presence of hypoxia response elements (HREs), serving as binding sites for HIF-1α. Intriguingly, experimental evidence demonstrated that SpHIF-1α actively promotes SpLDH transcription through these HREs. To further elucidate the functional significance of SpHIF-1α, targeted silencing was employed, resulting in a substantial reduction in SpLDH expression, activity, and lactate concentrations in MCDV-1-infected mud crabs. Notably, SpHIF-1α-silenced mud crabs exhibited higher survival rates and lower viral loads in hepatopancreas tissues following MCDV-1 infection. These results highlight the critical role of SpHIF-1α in MCDV-1 pathogenesis by regulating LDH gene dynamics, providing valuable insights into the molecular mechanisms underlying the virus-host interaction.
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Braquiúros , Dicistroviridae , Animais , Braquiúros/metabolismo , Ácido Láctico/metabolismo , L-Lactato Desidrogenase/genética , L-Lactato Desidrogenase/metabolismo , Fator 1 Induzível por Hipóxia/genética , Fator 1 Induzível por Hipóxia/metabolismo , HipóxiaRESUMO
Glutaredoxin (Grx) is a glutathione-dependent oxidoreductase that plays a key role in antioxidant defense. In this study, a novel Grx2 gene (SpGrx2) was identified from the mud crab Scylla paramamosain, which consists of a 196 bp 5' untranslated region, a 357 bp open reading frame, and a 964 bp 3' untranslated region. The putative SpGrx2 protein has a typical single Grx domain with the active center sequence C-P-Y-C. The expression analysis revealed that the SpGrx2 mRNA was most abundant in the gill, followed by the stomach and hemocytes. Both mud crab dicistrovirus-1 and Vibrioparahaemolyticus infection as well as hypoxia could differentially induce the expression of SpGrx2. Furthermore, silencing SpGrx2 in vivo affected the expression of a series of antioxidant-related genes after hypoxia treatment. Additionally, SpGrx2 overexpression significantly increased the total antioxidant capacity of Drosophila Schneider 2 cells after hypoxia, resulting in a reduction of reactive oxygen species and malondialdehyde content. The subcellular localization results indicated that SpGrx2 was localized in both the cytoplasm and the nucleus of Drosophila Schneider 2 cells. These results indicate that SpGrx2 plays a crucial role as an antioxidant enzyme in the defense system of mud crabs against hypoxia and pathogen challenge.
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Proteínas de Artrópodes , Braquiúros , Glutarredoxinas , Animais , Braquiúros/imunologia , Braquiúros/microbiologia , Glutarredoxinas/química , Glutarredoxinas/genética , Glutarredoxinas/metabolismo , Proteínas de Artrópodes/metabolismo , Drosophila , Especificidade de Órgãos , Sequência de Bases , Sequência de Aminoácidos , Oxigênio/metabolismo , Transcriptoma , Oxirredutases/metabolismo , Clonagem Molecular , Linhagem CelularRESUMO
Cadmium is one of hazardous pollutants that has a great threat to aquatic organisms and ecosystems. The intestine plays important roles in barrier function and immunity to defend against environmental stress. However, whether cadmium exposure caused the intestine injury is not well studied. Thus, the aim of this study was to explore the potential mechanisms of cadmium toxicity in the intestine of mud crab (Scylla paramamosain) via physiological, histological, microbial community, and transcriptional analyses. Mud crabs were exposed to 0, 0.01, and 0.125 mg/L cadmium. After a 21-day of cadmium exposure, 0.125 mg/L cadmium caused intestine damaged by decreasing superoxide dismutase and catalase activities, and increasing hydrogen peroxide and malondialdehyde levels. Integrated biological index analysis confirmed that the toxicity of cadmium exhibited a concentration-dependent manner. Comparative transcriptional analyses showed that the up-regulations of several genes associated with heat shock proteins, detoxification and anti-oxidant defense, and two key signaling pathways (PI3k-Akt and apoptosis) revealed an adaptive response mechanism against cadmium exposure. Transcriptomic analysis also suggested that cadmium exposure disturbed the expression of ion transport and immune-related genes, indicating that it has negative effects on the immune functions of the mud crab. Furthermore, the intestinal microbial diversity and composition were significantly influenced by cadmium exposure. The abundance of the dominant phyla Fusobacteria and Bacteroidetes significantly changed after cadmium exposure. KEGG pathway analysis demonstrated that cadmium exposure could change energy metabolism and environmental information processing. Overall, we concluded that excessive cadmium exposure could be potentially exerted adverse effects to the mud crab health by inducing oxidative damage, decreasing immune system, disrupting metabolic function, and altering intestinal microbial composition. These results provided a novel insight into the mechanism of cadmium toxicity on crustaceans.
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Braquiúros , Microbiota , Animais , Transcriptoma , Braquiúros/metabolismo , Cádmio/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Estresse Oxidativo , Antioxidantes/metabolismo , IntestinosRESUMO
Prolyl hydroxylase 2 (PHD2) is the key oxygen sensor that regulates the stability of the hypoxia-inducible factor -1α (HIF-1α). In this study, a novel PHD2 gene from the mud crab Scylla paramamosain, named SpPHD2, was cloned and identified. The full-length transcript of SpPHD2 was found to be 1926 bp, consisting of a 333 bp 5' untranslated region, a 1239 bp open reading frame, and a 354 bp 3' untranslated region. The putative SpPHD2 protein contained a Prolyl 4-hydroxylase alpha subunit homologues (P4Hc) domain in the C-terminal and a Myeloid translocation protein 8, Nervy, and DEAF-1 (MYND)-type zinc finger (zf-MYND) domain in the N-terminal. The mRNA expression of SpPHD2 was found to be widely distributed across all examined tissues. Additionally, the subcellular localization results indicated that the SpPHD2 protein was mainly localized in the cytoplasm. The in vivo silencing of SpPHD2 resulted in the upregulation of SpHIF-1α and a series of downstream genes involved in the HIF-1 pathway, while SpPHD2 overexpression in vitro dose-dependently reduced SpHIF-1α transcriptional activity, indicating that SpPHD2 plays a crucial role in SpHIF-1α regulation. Interestingly, the expression of SpPHD2 increased under hypoxic conditions, which was further inhibited by SpHIF-1α interference. Moreover, four hypoxia response elements were identified in the SpPHD2 promoter, suggesting that a feedback loop exists between SpPHD2 and SpHIF-1α under hypoxia. Taken together, these results provided new insights into the regulation of SpPHD2 in response to hypoxia in S. paramamosain.