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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 H

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.

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.

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.

Aqueous batteries that use metal anodes exhibit maximum anodic capacity, whereas the energy density is still unsatisfactory partially due to the high redox potential of the metal anode. Current metal anodes are plagued by the dilemma that the redox potential of Zn is not low enough, whereas Al, Mg, and others with excessively low redox potential cannot work properly in aqueous electrolytes. Mn metal with a suitably low redox potential is a promising candidate, which was rarely explored before. Here, we report a rechargeable aqueous Mn-metal battery enabled by a well-designed electrolyte and robust inorganic-organic interfaces. The inorganic Sn-based interface with a bottom-up microstructure was constructed to preliminarily suppress water decomposition. With this bubble-free interface, the organic interface can be formed via an esterification reaction of sucrose triggered by acyl chloride in the electrolyte, generating a dense physical shield that isolates water while permitting Mn

Metal-organic frameworks (MOFs) have drawn growing attention as promising electrode candidates for rechargeable batteries. However, most studies focus on the direct use of MOF electrodes without any modification. Post-synthetic modification, a judicious tool to modify targeted properties of MOFs, has been long-neglected in the field of batteries. Herein, crystal-facet engineering is proposed to design MOF-based electrodes with high capacity and fast electrochemical kinetics. We found that a thermally-modified strategy can regulate the dominant exposed facet of Ni-based MOF (PFC-8). With the optimally exposed facets, the battery exhibited admirable rate capability (139.4 mAh g

Mud crab (Scylla paramamosain) is an important economic species in China. Vibrio parahaemolyticus infection have caused a great economic loss in mud crab farming. The mechanism involved in the immune responses of mud crab to V. parahaemolyticus is unclear. In this study, the physiological and immune response to V. parahaemolyticus infection were investigated in S. paramamosain. The results showed that V. parahaemolyticus infection decreased total hemocyte counts, led to cytological damage, and caused high mortality. Transcriptome analysis showed that 1327 differentially expressed genes (DEGs), including 809 up-regulated and 518 down-regulated ones, were obtained after V. parahaemolyticus challenge. These DEGs were mainly involved in the immune response and infectious disease. Additionally, transcriptome analysis revealed that Toll, immune deficiency (IMD), and prophenoloxidase signalling pathways played essential roles in antibacterial immunity against V. parahaemolyticus infection in mud crab.

Vitamin D3 (VD3) has been shown to modulate the innate immune response in mammals but this has been rarely reported in fish. The current study found that increasing dietary VD3 content can reduce the density of yellow to dark brown pigmented macrophage aggregates (PMAs) in the spleens of yellow catfish infected with Edwardsiella ictaluri. The results of next-generation sequencing showed that a high dose of dietary VD3 (16,600 IU/kg) mainly affected the splenic immune response during Edwardsiella ictaluri infection via negative regulation of 'NF-κΒ transcription factor activity', 'NIK/NF-κΒ signaling' and the 'i-kappab kinase/NF-κΒ signaling' pathways. Follow-up qPCR showed that dietary VD3 increased the expression of NF-κΒ inhibitor iκb-α, decreased the expression of nf-κb p65, il-6, il1-β and tnf-α, and down-regulated the expression of nik, ikks and nf-κb p52 in the NIK/NF-kappaB signaling pathway. The above results indicate that dietary VD3 can modulate the splenic innate immune response of yellow catfish after Edwardsiella ictaluri infection by inhibiting the NF-κB activation signaling pathways.

Transmissible spongiform encephalopathies (TSEs) are epidemic neurodegenerative diseases caused by prion proteins, in particular, they are induced by misfolded prion proteins (PrPSc). PrPSc tend to aggregate into insoluble amyloid prion fibrils (fPrPWT), resulting in apoptosis of neuron cells and sequential neurodegeneration. Previous studies indicate that microglia cells play an important role in the innate immune system, and that these cells have good neuroprotection and delay the onset of TSEs. However, microglia can be a double-sided blade. For example, both Cu2+ and Mn2+ can induce microglia activation and secrete many inflammatory cytokines that are fatal to neuron cells. Unfortunately, PrP have cation binding sites at the N-terminus. When PrPSc accumulate during microglial phagocytosis, microglia may change the phenotype to secrete pro-inflammation cytokines, which increases the severity of the disease. Some studies have revealed an increase in the concentration of Mn2+ in the brains of patients. In this study, we treated microglia with fPrPWT and cations and determined I&kappa, B&alpha, and IL-1&beta, expression by Western blotting and quantitative polymerase chain reaction. The results showed that Mn&ndash, fPrPWT decreased I&kappa, levels and dramatically increased IL-1&beta, mRNA expression. In addition, competing binding between Cu2+ and Mn2+ can decrease the effect of Mn&ndash, fPrPWT on I&kappa, The effects of divalent cations and fPrPWT in microglia inflammation are also discussed.

Atopic dermatitis (AD) is a chronic skin inflammatory disease characterized by disequilibrium between Th1/Th2 lymphocytes.This study aimed to investigate the therapeutic effect of HP-NAP in AD mice model.The model of AD was built with oxazolone (OXA) in BALB/c mice, then HP-NAP was used to treat AD by intraperitoneal injection. Ear thickness was measured by a digital thickness gauge. The ears tissues were collected and subjected to hematoxylin-eosin (HE) and toluidine blue (TB) staining. The mRNA expression levels of inflammatory cytokines (IL-1β, IL-5, IL-6, and TNF-α) in ear tissue were measured using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The secretion of IgE, IL-4, and IFN-γ was measured by enzyme-linked immunosorbent assay (ELISA).Treatment with HP-NAP successfully alleviated the symptoms of AD, such as erythema, horny substance, and swelling. The infiltration of lymphocytes and mast cells were significantly reduced following HP-NAP therapy. The secretion of IgE and IL-4 was significantly attenuated following treatment with HP-NAP. Additionally, HP-NAP observably downregulated inflammatory cytokine expression (e.g. IL-1β, IL-5, IL-6, and TNF-α) in ear tissues.Taken together, our results showed that HP-NAP possessed the potential to be a novel immunomodulatory candidate drug against AD.

Cadmium is one of the most common heavy metal pollutants in the aquatic environment. Mud crab (Scylla paramamosain) is considered a model organism to monitor the impact of heavy metals. However, knowledge about toxicological mechanism of cadmium in crustaceans still remains limited. In this study, mud crabs were exposed to different concentrations of cadmium (0, 1.25, 2.5, 5 and 10 mg/L) for 72 h. Cadmium exposure significantly decreased superoxide dismutase (SOD) activity, catalase (CAT) activity and total antioxidative capacity (T-AOC), and significantly increased malondialdehyde (MDA) and H2O2 levels. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) activity significantly increased after cadmium exposure. Moreover, integrated biological responses version 2 (IBRv2) analysis suggested that cadmium exposure exerted stronger toxicity on mud crab. Furthermore, oxidative stress induced by cadmium exposure could decrease total hemocyte count (THC), interrupt Ca2+ homeostasis, and lead to cytological damage. Cadmium exposure induced DNA damage, which activated DNA damage response signaling ATR-CHK1-p53 pathway. Our results also showed that cadmium exposure significantly increased the apoptosis and caspase-3 mRNA levels, which implied that cadmium induced apoptosis through a caspase-3 pathway.

MYB transcription factors (TFs) play important roles in the plant's response to abiotic stress. In this study, we cloned a novel MYB TF gene from Vaccinium corymbosum (blueberry) using rapid amplification of cDNA ends (RACE). The cDNA contained a 798-bp open reading frame that encodes a 265-amino acid protein. VcMYB4a possessed a C2/EAR-repressor motif domain and phylogenetic analysis showed that it clustered into a subgroup 4 with six Arabidopsis thaliana MYBs. Quantitative RT-PCR analysis demonstrated that VcMYB4a expression was downregulated by salt, drought, and cold treatment, but was induced by freezing and heat. Overexpression of VcMYB4a in blueberry callus enhanced sensitivity to salt, drought, cold, freezing, and heat stress. These results indicate that VcMYB4a may be an important repressor of abiotic stress in blueberry.

The immune and physiological responses of mud crab (Scylla paramamosain) under air exposure were studied. The results showed that air exposure increased plasma activities of AST, ALT, ALP. There was a significant increase in glucose (GLU) and malondialdehyde (MDA) levels after air exposure. The transcript levels of SOD, CAT, HSP90, HSP70, p53, and hypoxia-inducible factor-1 (HIF-1) were induced by air exposure. Furthermore, caspase-3 transcript significantly increased at 48 and 72 h, while it significantly decreased at 96 h and 120 h under air exposure. These results suggested that oxidative stress occurred in the prolonged period of air exposure. HIF-1 and p53 signaling pathways played an important role under air exposure.

The present study was aimed to investigate the low temperature toxicity and its protection by taurine in pufferfish. The experimental basal diets supplemented with taurine at the rates of 250 (control), 550, 850, 1140, 1430, 1740 mg kg−1 were fed to fish for 8 weeks. The results showed that fish fed diet with taurine had significantly improved weight gain and specific growth rate. After the feeding trial, the fish were then exposed to low temperature stress. The results showed that low temperature stress could induce reactive oxygen species (ROS) generation, disturb the cytoplasm Ca2+ homeostasis, and lead to oxidative stress and apoptosis. Compared with the control group, dietary taurine supplementation groups increased antioxidant enzyme genes such as manganese superoxide dismutase (Mn-SOD), glutathione peroxidase (GPx), glutathione reductase (GR) and catalase (CAT), heat shock proteins (HSP70) and complement C3 (C3) mRNA levels under low temperature stress. Meanwhile, dietary taurine supplementation groups reduced ROS generation, and stabilized the cytoplasm Ca2+ under low temperature stress. Furthermore, dietary taurine supplementation groups reduced apoptosis via decreasing caspase-3 activity. This is the first report to demonstrate the mechanisms of taurine against low temperature stress in fish.

This study was conducted to determine the effects of vitamin E on growth performance, biochemical parameters, and antioxidant capacity of pufferfish (Takifugu obscurus) exposed to ammonia stress. The experimental basal diets supplemented with vitamin E at the rates of 2.31 (control), 21.84, 40.23, 83.64, 158.93, and 311.64 mg kg−1 dry weight were fed to fish for 60 days. After the feeding trial, the fish were exposed to 100 mg L−1 ammonia-nitrogen for 48 h. The results shown that the vitamin E group significantly improved weight gain, specific growth rate, and the expression levels of growth hormone receptors and insulin-like growth factor. Fish fed with the vitamin E-supplemented diets could increase plasma alkaline phosphatase activities and decrease plasma glutamicoxalacetic transaminase and glutamic-pyruvic transaminase activities. The relative expression levels of heat shock proteins (40.23–311.64 mg kg−1 vitamin E diet group), manganese superoxide dismutase (83.64–158.93 mg kg−1 vitamin E diet group), catalase (40.23–311.64 mg kg−1 vitamin E diet group), and glutathione reductase (40.23–311.64 mg kg−1 vitamin E diet group) were upregulated. On the other hand, the decreased level of reactive oxygen species (ROS) was observed in the 83.64–311.64 mg kg−1 vitamin E additive group. These results showed that vitamin E might have a potentially useful role as an effective antioxidant to improve resistance in pufferfish.

The tumor suppressor protein p53 plays important roles in DNA repair, cell cycle and genetic stability. In the present study, a p53 gene in the mud crab (Scylla paramamosain) (designated as Sp-53) was identified and characterized. The open reading frame of Sp-53 was comprised a 1383 bp, which encoded a putative protein of 460 amino acids. Sp-53 is expressed in all examined tissues, with the highest expression in hepatopancreas and hemocytes. Vibrio parahaemolyticus infection induced oxidative stress, and led to DNA damage. The Sp-53 transcriptions in hepatopancreas were significantly up-regulated after V. parahaemolyticus infection. RNA interference (RNAi) experiment was used to understand the roles of Sp-53 in response to V. parahaemolyticus infection. Knocking down Sp-53 in vivo significantly reduced the expression of the Mn-SOD, Gpx3 and caspase 3 after V. parahaemolyticus infection. Moreover, the mortality of mud crabs and DNA damage in Sp-53-silenced mud crab challenged with V. parahaemolyticus were significantly higher than those in the control group. All these results suggested that Sp-53 played an important role in responses to V. parahaemolyticus infection through its participation in regulation of antioxidant defense, DNA repair and apoptosis.

Mud crab (Scylla paramamosain) is an economically important cultured species in China. Hypoxia is a major environmental stressor during mud crab culture. In the present study, we investigated the oxidative stress and transcriptome changes in the gills of mud crab after intermediate hypoxia stress with dissolved oxygen (DO) 3.0 ± 0.2 mg/L (named as “DO3”) and acute hypoxia stress with DO 1.0 ± 0.2 mg/L (named as “DO1”) for 0, 3, 6, 12 and 24 h. The superoxide dismutase (SOD) activity of DO1 increased significantly at 3, 6 and 24 h after hypoxia stress, while SOD activity of DO3 increased significantly at 6 and 24 h. The total antioxidant capacity (T-AOC) increased significantly at 6, 12 and 24 h after hypoxia stress. The malondialdehyde (MDA) concentration of DO1 increased significantly at 6, 12 and 24 h after hypoxia stress, while MDA concentration of DO3 only increased significantly at 6 h. The lactate dehydrogenase (LDH) activity of DO1 increased significantly at 3, 6, 12 and 24 h after hypoxia stress, while LDH activity of DO3 increased significantly at 12 and 24 h. Transcriptomic analysis was conducted at 24 h of gill tissues after hypoxia stress. A total of 1052 differentially expressed genes (DEGs) were obtained, including 394 DEGs between DO1 and DO3, 481 DEGs between DO1 and control group, 177 DEGs between DO3 and control group. DEGs were enriched in the pathways related to metabolism, immune functions, ion transport, and signal transduction. Transcriptional analysis showed that glycolysis and tricarboxylic acid cycle genes were the key factors in regulating the adaptation of mud crab to hypoxia stress.

Apoptosis plays a crucial role in many biological processes, including development, cellular homeostasis, and immune responses. Bax inhibitor-1 (BI-1) is an anti-apoptotic protein that protects cells from endoplasmic reticulum stress-induced apoptosis. In this study, a BI-1 gene from the pufferfish Takifugu obscurus (Pf-BI-1) was identified and characterized. The full length of Pf-BI-1 cDNA was 1387 bp, including a 5'-UTR of 82 bp, a 3'-UTR of 591 bp containing a poly-(A) tail, and an open reading frame (ORF) of 714 bp that encodes a polypeptide of 237 amino acids. Pf-BI-1 was ubiquitously expressed in various tissues, with the highest expression levels in the blood, brain, and gill. The expression of Pf-BI-1 was up-regulated in a time-dependent manner after heat shock stress, ammonia stress, and bacterial challenge. Intracellular localization revealed that Pf-BI-1 was primarily localized in the cell cytoplasm. Furthermore, over-expression of Pf-BI-1 could active NF-кB reporter genes in HeLa cells. These results indicated that Pf-BI-1 may be involved in the apoptosis and immunity process against ambient stressors in pufferfish.

The tumor suppressor protein p53 plays a critical role in cell cycle, apoptosis and DNA repair. In this study, the full-length pufferfish p53 (Pf-p53) was obtained, containing an open reading frame of 1095 bp, a 5′UTR of 157 bp and a 3′UTR of 285 bp with a poly (A) tail. The Pf-p53 encoded a polypeptide of 364 amino acids with a theoretical isoelectric point of 8.03 and predicted molecular weight of 40.6 kDa. Pf-p53 was ubiquitously expressed in various tissues with a high-level expression in kidney, liver and gill. Vibrio alginolyticus challenge could induce ROS production and disrupt Ca 2+ homeostasis, subsequently leading to the induction of DNA damage and apoptosis, while the Vibrio alginolyticus -induced oxidative stress can also increase the non-specific immunity. The pufferfish challenged with Vibrio alginolyticus showed a sharp increase of Pf-p53 transcript in liver. Subcellular localization analysis revealed that Pf-p53 was primarily localized in nucleus. Furthermore, overexpression of Pf-p53 in Hela cells could inhibit cell proliferation and the transcriptional activities of the NF-ĸB promoter. Taken together, our results indicated that Pf-p53 may play an important role in the immune response to Vibrio alginolyticus challenge.

RNA binding proteins (RBPs) regulate gene expression by controlling mRNA export, translation, and stability. When altered, some RBPs allow cancer cells to grow, survive, and metastasize. Cold-inducible RNA binding protein (CIRP) is overexpressed in a subset of breast cancers, induces proliferation in breast cancer cell lines, and inhibits apoptosis. Although studies have begun to examine the role of CIRP in breast and other cancers, its role in normal breast development has not been assessed. We generated a transgenic mouse model overexpressing human CIRP in the mammary epithelium to ask if it plays a role in mammary gland development. Effects of CIRP overexpression on mammary gland morphology, cell proliferation, and apoptosis were studied from puberty through pregnancy, lactation and weaning. There were no gross effects on mammary gland morphology as shown by whole mounts. Immunohistochemistry for the proliferation marker Ki67 showed decreased proliferation during the lactational switch (the transition from pregnancy to lactation) in mammary glands from CIRP transgenic mice. Two markers of apoptosis showed that the transgene did not affect apoptosis during mammary gland involution. These results suggest a potential in vivo function in suppressing proliferation during a specific developmental transition.

The aquatic microbial community is sensitive to environmental change; however, the impacts of those changes combined with disease outbreaks affecting S. paramamosain are unknown. Thus, from March to October, we explored the interaction between aquacultural environmental conditions and microbial composition and function in open-air aquaculture ponds containing S. paramamosain in Southern China. The microbial community structure was significantly positively correlated with microbial community function. The environment variables such as temperature and salinity during May and June changed more quickly compared with other periods, resulting changes in the structure and function of the microbial community affected S. paramamosain survivability, with higher crab mortality observed from May to June compared with other periods. These included changes in the relative abundance of Microtrichales, Synechococcales, Rhodobacterales, Chitinophagales, and SAR11_clade, and corresponding functions associated with glycolysis and/or gluconeogenesis, porphyrin and chlorophyll metabolism, photosynthetic proteins, and transcription factors. These changes could impact S. paramamosain mortality and be used to evaluate the health status of the ponds. Though the environment variables during July~October changed slowly comparing to May and June, the ponds microflora changed which benefit S. paramamosain survivability with correspondingly low S. paramamosain mortality. Therefore, rapid environmental change alters the structure and function of the aquatic microflora, increasing S. paramamosain mortality.

Glutathione S-transferase (GST) plays important roles in cellular detoxification and antioxidant defense. A Mu-type glutathione S-transferase (designated as SpMu-GST) was obtained from the mud crab Scylla paramamosain. The open reading frame of SpMu-GST was comprised a 690 bp, which encoded a putative protein of 229 amino acids. Quantitative real-time PCR (qRT-PCR) revealed that the SpMu-GST mRNA was expressed in all examined tissues, with highest expression in hepatopancreas. During ammonia exposure, the SpMu-GST transcriptions in hepatopancreas and gill were significantly up-regulated at early exposure time. Moreover, RNA interference (RNAi) experiment was designed to understand the roles of SpMu-GST under ammonia exposure. Ammonia exposure reduced the levels of glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) and total antioxidative capacity (T-AOC), and increased the formation of malondialdehyde (MDA). After knockdown of the SpMu-GST level, GST activity and T-AOC were significantly decreased at some exposure time after ammonia exposure. However, the mortality of mud crabs and malondialdehyde (MDA) contents significantly increased under ammonia exposure. These results further suggested that SpMu-GST played a critical role in mud crab antioxidant defenses in response to environmental stress.

As the most abundant biological entities, viruses are major players in marine ecosystems. However, our knowledge about virus-host interactions and viral ecology in the deep sea remains very limited. In this study, a novel bacteriophage (designated as phage BVE2) infecting Bacillus cereus group bacteria, was isolated from deep-sea sediments. Phage BVE2 caused host lysis within 1.5 h after infection. However, the presence of two integrase-encoding genes in the BVE2 genome suggested that BVE2 may also follow a temperate strategy. The genome of phage BVE2 is approximately 20 kb in length and is predicted to encode 28 proteins. Genomic and phylogenetic analysis suggested that BVE2 is a highly mosaic phage that has inherited genetic features from Wbeta-like viruses, B. cereus prophages, and its host, suggesting that frequent horizontal gene transfer events occurred during its evolution. This study will help to reveal the evolutionary history of Wbeta-like viruses and improve our understanding of viral diversity and virus–host interactions in the deep sea.

The mud crab, Scylla paramamosain, is an economically-important crab in China. Air exposure is an important environmental stressor during mud crab culture and transportation. Adaptive mechanisms responding to air exposure in mud crabs are still poorly understood. In this study, mud crabs were exposed to air for 120 h. Air exposure decreased total hemocyte counts, led to cytological damage, and caused high mortality. Transcriptomic analysis was conducted at 0, 6 and 96 h after air exposure. A total of 3530 differentially expressed genes (DEGs) were identified. DEGs were mainly involved in the oxidative stress response, metabolism, cellular processes, signal transduction, and immune functions. Transcriptomic analysis also revealed that genes of glycolysis and of the tricarboxylic acid cycle were key factors in regulating the mud crab adaptation to air exposure.

Objective MicroRNAs (miRNAs) regulate prostate tumorigenesis and progression by involving different molecular pathways. In this study, we examined the role of miR-572 in prostate cancer (PCa). Methods The proliferation rates of LNCaP and PC-3 PCa cells were studied using MTT assays. Transwell migration and Matrigel invasion assays were performed to evaluate cell migration and invasion, respectively. Protein expression levels were examined using western blotting. Docetaxel-induced apoptosis was evaluated by Caspase-Glo3/7 assays. The putative miR-572 binding site in the phosphatase and tensin homolog (PTEN) 3ʹ untranslated region (3ʹ UTR) was assessed with dual-luciferase reporter assays. Additionally, miR-572 expression levels in human PCa tissues were examined by qRT-PCR assays. Results Upregulation of miR-572 promoted proliferation, migration, and invasion of PCa cells. Overexpression of miR-572 decreased sensitivity of PCa cells to docetaxel treatment by reducing docetaxel-induced apoptosis. MiR-572 can regulate migration and invasion in PCa cells. Furthermore, miR-572 could regulate expression of PTEN and p-AKT in PCa cells by directly binding to the PTEN 3ʹ UTR. MiR-572 expression levels were increased in human PCa tissues and associated with PCa stage. Conclusions miR-572 displayed essential roles in PCa tumor growth and its expression level may be used to predict docetaxel treatment in these tumors.

In this paper, the mechanics equation of the dynamic compaction machine arm frame is established based on the theory of statics, the arm lifting rope force is given, the influence of the wire rope force with relative position parameters changing is also discussed. The critical factors that affect the arm lifting rope force are identified. Compared the simulation and the test results, the major forces errors are less than 8%. The correctness of the theoretical study and simulation are verified. The research results could be used for the arm wire rope type selection of the dynamic compaction.

Nitrite is one of major environmental pollutants that can impact immunological parameters in aquatic organisms. In the present study, we investigated the effects of nitrite exposure on oxidative stress, DNA damage and apoptosis in mud crab (Scylla paramamosain). Mud crab were exposed to 0, 5, 10 and 15 mg L−1 nitrite for 72 h. These data showed that acid phosphatase (ACP) and alkaline phosphatase (ALP) activity significantly decreased in treatments with various concentrations of nitrite (5, 10 and 15 mg L−1) after 24 and 48 h, while the levels of nitric oxide (NO) significantly increased in these treatments. Nitrite exposure could suppress superoxide dismutase (SOD) and catalase (CAT) activity, and increase the formation of malondialdehyde (MDA) after 48 and 72 h of exposure. In addition, nitrite exposure decreased total haemocyte counts after 48 and 72 h of exposure. Cytological damage, DNA damage and apoptosis was observed obviously at 72 h after nitrite exposure. Moreover, nitrite exposure significantly induced the mRNA levels of phosphorylated Jun N-terminal kinases (JNK), and eventually activated p53 signaling and caspase-3. These results indicated that nitrite exposure could induce oxidative stress, which further caused DNA damage and apoptosis in mud crab. Our results will be helpful to understand the mechanism of nitrite toxicity on crustaceans.

The rapid emergence and dissemination of antibiotic resistance poses a threat to human health and to the marine environment. We have investigated the abundance and diversity of antibiotic resistance genes (ARGs) and of antibiotic-resistant bacteria (ARB), during the seedling period, rearing period, and harvesting period in seven marine fish cage-culture areas in Guangdong. Spatial and temporal variations of AGRs and ARB were also analyzed. Culture-based methods and quantitative PCR were used to detect ARB and ARGs. Bacterial resistance rates were no significantly different within farming periods. The proportion of antibiotic-resistant bacteria was extremely low (average on 1.15%), except for oxytetracycline-resistant bacteria (average on 34.15%). Vibrio was the most common ARB. Sul1, tetB, and ermB, had the highest relative abundance. The abundance of ARGs in the harvesting period was significant highest. The total abundance of ARGs was highest at Raoping and lowest at Dayawan and Liusha. Most ARGs were associated with opportunistic pathogens. The environmental factors effecting ARB and ARGs are complex, and no key factors were identified. This study provides a theoretical basis for assessing the harmfulness of ARGs and ARB to food safety and human health.

The current epidemiological situation of viral nervous necrosis virus (VNNV) on Hainan Island was investigated. A total of 490 hatchery-reared fish and 652 wild fish were sampled for VNNV detection from March 2013 to May 2014. Positive detection rates of 84.53% (153/181) and 0.97 % (3/309) were obtained in diseased and healthy hatchery-reared samples, respectively, by conventional RT-PCR. However, using more-sensitive nested RT-PCR, the positive detection rates in healthy hatchery-reared fish reached up to 64.08% (198/309), suggesting that asymptomatic VNNV carriers commonly exist among larvae and juveniles breeding on Hainan Island. In wild-fish samples, 2.6% (17/652) and 34.2% (223/652) positive detection rates were observed using RT-PCR and nested RT-PCR, respectively, indicating that wild fish may be a potential reservoir for VNNV. Phylogenetic analysis showed that all 52 VNNV isolates from cultured fish belong to the RGNNV genotype, but 2 out of 48 VNNV isolates from wild fish samples were found to be of the SJNNV genotype. This study is the first to confirm the existence of SJNNV-genotype VNNV in China. Golden pompano, an important fish species for culture, was selected as a fish model to investigate the optimal conditions for RGNNV disease progression in artificial infection experiments. The effects of temperature, salinity, and fish size were evaluated. Results showed that 28 °C and 20 ‰ are the optimal infection temperature and salinity, respectively, and golden pompano juveniles with small body sizes are more susceptible to RGNNV. These findings are highly consistent with those conditions involved in the natural outbreak of RGNNV.

RNA binding protein (RBPs) and microRNAs (miRNAs or miRs) are post-transcriptional regulators of gene expression that are implicated in development of cancers. Although their individual roles have been studied, the crosstalk between RBPs and miRNAs is under intense investigation. Here, we show that in breast cancer cells, cyclin E1 upregulation by the RBP HuR is through specific binding to regions in the cyclin E1 mRNA 3' untranslated region (3'UTR) containing U-rich elements. Similarly, miR-16 represses cyclin E1, dependent on its cognate binding sites in the cyclin E1 3'UTR. Evidence in the literature indicates that HuR can regulate miRNA expression and recruit or dissociate RNA-induced silencing complexes (RISC). Despite this, miR-16 and HuR do not affect the other’s expression level or binding to the cyclin E1 3'UTR. While HuR overexpression partially blocks miR-16 repression of a reporter mRNA containing the cyclin E1 3'UTR, it does not block miR-16 repression of endogenous cyclin E1 mRNA. In contrast, miR-16 blocks HuR-mediated upregulation of cyclin E1. Overall our results suggest that miR-16 can override HuR upregulation of cyclin E1 without affecting HuR expression or association with the cyclin E1 mRNA.

PID control and the velocity feedback control for ABS are studied of the wheel slip rate as controlled object. Comparison on these two kinds of control methods is analyzed. It is obtained that velocity feedback control and PID control exist much similarity in some extent, according to the hardware in the loop (HIL) simulation platform. However, the velocity feedback control has more obvious advantages than PID control in terms of the adaptability to different road conditions. It can maintain stable control of system dynamically under the changed road condition, leading to the braking distance shorter.

Based on the ABS model of a single wheel system, the traditional logic threshold, PID and fuzzy control method have been used to study the control system of commercial vehicle’s ABS. As the most important task of commercial vehicle’s ABS is to prevent the two front (steering) wheels from locking, the slip ratio should be controlled strictly in the safe range. A very short time of locking may cause the failure of steering, so control method which can keep slip ratio in the secure scope accurately can help vehicle avoid the danger of runaway. Set up the model on Simulink platform. Then, use the three control methods to control the ABS. By comparing and analyzing the processes and results under the control methods, find an appropriate way that can provide the most secure and effective control. From the perspective of processes (slip ratio changes), the simulations under the three control methods show the PID control gives the best slip ratio curve. The curve is almost coinciding with the ideal slip ratio curve. The oscillation of slip ratio given by fuzzy control is the most sever. From the point of view of the results (braking distance), the braking system with PID control has the shortest braking distance while with the fuzzy control has the longest. That is not to say fuzzy control is not advanced but the actual experiences difficult to get. And with the long response time, the fuzzy control is not suitable for commercial vehicle’s ABS. As a result and with the development and decreasing cost of PID controller, it is competitive in today’s ABS market.

Avian pathogenic Escherichia coli (APEC) induce septicemia in chickens by invading type II pneumocytes to breach the blood-air barrier. The virulence of APEC can be regulated by quorum sensing (QS). Andrographolide is a QS inhibitor of Pseudomonas aeruginosa (P. aeruginosa). Therefore, we investigate whether andrographolide inhibits the injury of chicken type II pneumocytes by avian pathogenic E. coli O78 (APEC-O78) by disrupting the bacterial QS system. The results showed that sub-MIC of andrographolide significantly reduced the release of lactate dehydrogenase (LDH), F-actin cytoskeleton polymerization, and the degree of the adherence to chicken type II pneumocytes induced by APEC-O78. Further, we found that andrographolide significantly decreased the autoinducer-2 (AI-2) activity and the expression of virulence factors of APEC-O78. These results suggest that andrographolide reduce the pathogenicity of APEC-O78 in chicken type II pneumocytes by interfering QS and decreasing virulence. These results provide new evidence for colibacillosis prevention methods in chickens.

The present study was conducted to investigate the effects of astaxanthin on growth performance, biochemical parameters, ROS production, and immune-related gene expressions of the pufferfish (Takifugu obscurus) under high temperature stress. The experimental basal diets supplemented with astaxanthin at the rates of 0 (control), 20, 40, 80, 160, and 320 mg kg−1 were fed to fish for 8 weeks. The results showed that the fish fed diet with 80, 160, and 320 mg kg−1 astaxanthin significantly improved weight gain and specific growth rate. Furthermore, fish fed the moderate dietary astaxanthin increased plasma alkaline phosphatase activities, and decrease plasma aspartate aminotransferase and alanine aminotransferase activities. After the feeding trial, the fish were exposed to high temperature stress for 48 h. The results shown that astaxanthin could suppress ROS production induced by high temperature stress. Meanwhile, compared with the control group, the astaxanthin groups increased SOD, CAT, and HSP70 mRNA levels under high temperature stress. These results showed that the basal diet supplemented with 80–320 mg kg−1 astaxanthin could enhance growth, nonspecific immune responses, and antioxidant defense system and improve resistance against high temperature stress in pufferfish.

The research achievement of strata movement law caused by subway construction and control standards was summarized and analyzed, and point out that this research on the laws of strata movement (or deformation) caused by subway construction mainly focused on three aspects. The first was research on the ground vertical settlement laws perpendicular to the tunnel excavation direction. The second was research on the ground vertical settlement laws parallel to the tunnel excavation direction. The third was research on the ground horizontal movement regularity perpendicular to the tunnel excavation direction. Through the analysis of the current research results, it is points out the shortages existing in the research, and deeply research in the future. It may provide the theoretical basis, also for practical engineering application provides a certain technical support.

Guava (Psidium guajava L.) leaf extracts have antiviral and antibacterial activity against shrimp pathogens such as yellow-head virus (YHV), white spot syndrome virus (WSSV), and Vibrio harveyi, which make it a potential water disinfectant for use in shrimp culture. In this study, the safety of guava leaf supplementation in shrimp was evaluated by studying its influence on growth and the non-specific immune response of Penaeus monodon. Six diets containing different levels of guava leaves (0% [basal diet], 0.025% [G1], 0.05% [G2], 0.1% [G3], 0.2% [G4], and 0.4% [G5]) were fed to groups of shrimp (1.576 ± 0.011 g body weight) in triplicate for 56 days. Growth performance (final body weight, WG, PWG, SGR) of shrimp fed guava leaf diets was significantly higher (P0.05) than that of shrimp fed on the basal diet. The G1 diet resulted in the highest body weight gain (308.44%), followed by the G2 (295.45%), G3 (283.05%), G5 (281.29%), G4 (276.11%), and finally the basal diet (214.58%). Survival of shrimp in the G1 diet group was higher than that of shrimp in the control and the other experimental groups; however, no statistical differences (P0.05) were found. Dietary supplementation with guava leaf improved the activities of prophenoloxidase (PO) and nitric oxide synthase (NOS) in serum, and of superoxide dismutase (SOD), acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LSZ) both in serum and hepatopancreas of shrimp. In the experimental groups, the activities of these enzymes followed a similar pattern of change; they increased initially at low levels of dietary supplementation and then decreased with increasing concentrations of dietary guava leaf. Serum PO and SOD activities in shrimp fed the G1 diet reached 7.50 U ml(-1) and 178.33 U ml(-1), respectively, with PO activity being significantly higher than in controls. In shrimp fed the G1 diet, SOD, ACP, and AKP activities in hepatopancreas were significantly higher than in the controls, reaching 57.32 U g(-1), 23.28 U g(-1), and 19.35 U g(-1) protein, respectively. The highest activities of serum ACP, AKP, LSZ, and of hepatopancreas LSZ, were observed in the G3 diet group. Total nitric oxide synthase (TNOS) activity was highest (64.80 U ml(-1)) in the G4 diet group, which was significantly higher than that observed in the control group. These results suggest that dietary guava leaf supplementation could enhance the growth performance and non-specific immune response of P. monodon. Therefore, guava leaf is considered safe for use as a water disinfectant in shrimp culture.

Through research of vehicle dynamic feature, analysis and research of the problem of driving wandering by user retroactions, the article put forward some test and evaluation methods of driving wandering. And the article is discussed about main reasons and solutions of driving wandering in-depth.

Bottom-business multi-story masonry structure is widely used in small and middle towns in the southward in China. In the downtown of Beichuan county which affected by Wenchuan earthquake, more than 80% of this kind of building collapsed. But the Apartment of Beichuan Telecommunication Bureau behaved well earthquake resistant capacity with a moderate damage in the earthquake. The obvious difference between this building and others is the setting of winged columns in the front longitudinal wall of the first floor. For proving the influence of these members in the structure seismic capacity, the earthquake simulation shaking table test of 2 1/5 reduced scale models were designed and carried out. The models dynamic response, acceleration, displacement and strain were measured and collected, that model with winged columns behave well was testified. And the reinforcement method of balancing stiffness and increasing ductility is put forward. The results provide a foundation for the retrofitting design of the existing houses.

Ginsenoside Rh2, an active component of ginseng, exhibits immunoregulatory and anti-inflammatory properties. Rh2-B1, a sulfated derivative, was prepared to enhance its water solubility. We studied the effect of Rh2-B1 on CTLL-2, a CD8⁺ cytotoxic T cell line that was known for protecting against viral infection.We aimed to investigate the effect of Rh2-B1 on interferon (IFN)-γ production and cell proliferation and its possible mechanism.Enzyme-linked immunosorbent assay (ELISA) was employed to analyze the IFN-γ concentration of the whole blood and the supernatant of CTLL-2 cell culture. Cell proliferation assay was conducted using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Western blots were used to evaluate changes in signal transduction pathways in CTLL-2 cells.Rh2-B1 was able to enhance IFN-γ production from whole blood culture of Balb/c mice. We then evaluated the effect of Rh2-B1 on a cytotoxic T cell line, CTLL-2 for cell proliferation, IFN-γ production and its molecular mechanism. Rh2-B1 promoted cell proliferation and IFN-γ production of CTLL-2 cells. It also induced activation of p38 mitogen-activated protein kinase (MAPK) and extracellular-signal-regulated kinases (ERK), but inhibited p56 Lck and transducer and activator of transcription 5 (STAT5) expression. The effect was blocked by the specific p38 MAPK inhibitor SB203580 and ERK inhibitor U0126.Rh2-B1 could stimulate cell proliferation and IFN-γ production by activating the p38 MAPK- and ERK-dependent signaling pathways in cytotoxic T cells. This may be a novel medicine for treatment of viral infections.

A parametric FEM analysis method is introduced for the design of disc brakes. The basic process of parametric FEM modeling is summarized, the FEM model of disc brakes is established. Finally, the parametric FEM strength and stiffness analysis are conducted for the typical disc brake assembly.

Shrinkage cracking is the direct cause which leads to the deterioration of durability and integrity of concrete. In this paper, ring-test methods are used to study the early age cracking characterization of compound mineral admixtures concrete which includes steel slag, blast furnace slag and fly ash. The effect of compound admixtures on cracking characterization of concrete is discussed, restraint stress caused by concrete shrinkage and cracking age are analyzed. Through the comprehensive consideration of the free shrinkage strain, creep, restriction factors on the effect of the cracking of concrete specimens. The results show that suitable proportion mineral admixtures can be used to reduce cracking in concrete. The risk of cracking can be estimated for compound admixtures concrete, so that it will improve the durability of concrete structures.

Low temperature is an important environmental factor in aquaculture farming that affects the survival and growth of organisms. In the present study, we investigated the effects of low temperature on biochemical parameters, oxidative stress and apoptosis in pufferfish. In the stress group, water temperature decreased from 25 °C to 13 °C at a rate of 1 °C/1 h. Fish blood and liver were collected to assay biochemical parameters, oxidative stress and expression of genes at 25 °C, 21 °C, 17 °C, 13 °C and 13 °C for 24 h. The results showed that low temperature could decrease total blood cell count, inhibit cell viability, and subsequently lead to DNA damage. Biochemical parameters such as plasma protein and ALP significantly declined in fish under low temperature, while a significant increase in AST, ALT, LDH and glucose was observed. The gene expression of antioxidant enzymes (SOD and CAT), HSP90 and C3 were induced by low temperature stress. Furthermore, the gene expression of apoptotic related genes including P53, caspase-9 and caspase-3 were up-regulated, suggesting that caspase-dependent pathway could play important roles in low temperature-induced apoptosis in fish. This study may provide baseline information about how cold stress affects the physiological responses and apoptosis in fish.

In the past, when need to draw one model of brake parts, users have to use the CAD software and make it one step by step, which wastes a lot of time and energy because of the repetitive artificially drawing. In order to improve the efficiency of drawing the models of brake parts, the paper builds one brake parts library by using the Visual C++6.0 and doing secondary development on the CATIA. Through one example, the paper proves the feasibility of building brake parts library.

Chemokines are small, secreted cytokine peptides known principally for their ability to induce migration and activation of leukocyte populations and regulate the immune response mechanisms. The cobia (Rachycentron canadum), a marine finfish species, has a great potential for net cage aquaculture in the South China Sea. We isolated and characterized a CC chemokine cDNA from cobia-designated RcCC2. Its cDNA is 783 bp in length and encodes a putative protein of 110 amino acids. Homology and phylogenetic analysis revealed that the RcCC2 gene, which contains four conserved cysteine residues, shares a high degree of similarity with other known CC chemokine sequences and is closest to the CCL19/21 clade. The mRNA of RcCC2 is expressed constitutively in all tested tissues, including gill, liver, muscle, spleen, kidney, head kidney, skin, brain, stomach, intestine and heart, but not blood, with the highest level of expression in gill and liver. The reverse transcription quantitative polymerase chain reaction was used to examine the expression of the RcCC2 gene in immune-related tissues, including head kidney, spleen and liver, following intraperitoneal injection of the viral mimic polyriboinosinic polyribocytidylic acid, formalin-killed Vibrio carchariae (bacterial vaccine) and phosphate-buffered saline as a control. RcCC2 gene expression was up-regulated differentially in head kidney, spleen and liver during 12 h after challenge. These results indicate that the RcCC2 gene is inducible and is involved in immune responses, suggesting RcCC2 has an important role in the early stage of viral and bacterial infections.

A hybrid coupling beam is developed in this study to render the reinforced concrete (RC) shear wall structure a higher seismic performance. As the first protection from earthquake, the coupling beam is able to dissipate more energy when combined in series with a metallic damper. The strength of the metallic damper is carefully selected so that the RC part of the coupling beam remains elastic, while all plasticity goes into the metallic damper. This mechanism protects the RC part from seismic damage. And the metallic damper can be quickly replaced once it is damaged. This significantly enhances the reparability of entire structure, making it possible to be immediately functioned after earthquakes. The stiffness of the hybrid coupling beam is similar as the traditional RC coupling beam, so that the dynamic characteristics are almost unchanged. In this study, one hybrid coupling beam and one traditional RC beam are experimentally examined. The seismic dissipation capability of the hybrid beam is approximately twice of the traditional beam, and the damage of the RC part of the hybrid beam is very limited.

Considering the importance of vehicle active safety system, this article designs the manageable man-machine interactive software that can help engineers to design the vehicle active safety system. Engineers can interact with the software and involve in most processes of active safety system design through the visual software. And the software can improve design efficiency, shorten the design cycle, and reduce design costs. Through the successful development of the software, the paper shows the important role VC++ plays in vehicle active safety system design and this will be instructive.