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Edwardsiella tarda is one of the most harmful bacterial pathogens for aquaculture flatfish. After artificial infection of 47 Japanese flounder (Paralichthys olivaceus) families, resistant and susceptible families were identified in this study. High-throughput sequencing was performed on the liver transcriptome of uninfected groups (PoRU and PoSU) and infected groups (PoRC and PoSC). Through assembly and annotation, a total of 3012 and 1386 differentially expressed genes (DEGs) were identified in PoRU vs. PoSU and PoRC vs. PoSC. The significant enrichment pathways between PoRU and PoSU were mainly in metabolic and biosynthesis pathways. A total of thirty dominant enrichment pathways between PoRC and PoSC mainly focused on some immune-related pathways, including the hematopoietic cell lineage, cytokine-cytokine receptor interaction, complement and coagulation cascades, antigen processing and presentation, the intestinal immune network for immunoglobulin A (IgA) production and T/B cell receptor signaling pathway. Under the protein-protein interaction (PPI) analysis, hub genes, including CD molecules, complement component factors and chemokines, were identified in the network, and 16 core genes were differentially expressed in resistant and sustainable families in quantitative polymerase chain reaction (qPCR) validation. This study represents the first transcriptome analysis based on resistant and susceptible families and provides resistant genes to understand the potential molecular mechanisms of antibacterial function in marine fish. The results obtained in this study provide crucial information on gene markers for resistant breeding of Japanese flounder.

Zirconia-coated Cr2O3 aggregates synthesized by mixing ZrO2 powders and Cr2O3 aggregates with a phenolic resin binder followed by thermosetting treatment were employed as modified Cr2O3 aggregates to obtain Cr2O3–Al2O3–ZrO2 bricks (high-chromia bricks). The elastic modulus (E) and cold modulus of rupture (CMOR) of these high-chromia bricks before and after thermal shock cycles were systematically investigated, and the residual ratios of CMOR and E were calculated. The thermal shock resistance of the high-chromia bricks was significantly improved by the factor of modification of Cr2O3 aggregates. The mechanism of the improved thermal shock resistance of these high-chromia bricks was studied via microstructure analysis, and the crack propagation behavior was analyzed by scanning electron microscopy (SEM). The fracture work (γWOF), thermal shock damage factor (R′′′′), and thermal stress crack stability parameter (Rst) were measured and calculated using the wedge splitting test (WST). The results indicate that the porous ZrO2 coating layer wrapped the Cr2O3 aggregates, forming modified Cr2O3 aggregates that can increase crack deflection, free path of crack propagation, and fracture work, thus improving the thermal shock resistance of high-chromia bricks. The thermal shock resistance of the fabricated high-chromia bricks was highly correlated with the thickness of the ZrO2 coating layer surrounding the Cr2O3 aggregates. The variation trend of Rst is well consistent with the experimental results, which is suitable to evaluate the thermal shock resistance of high-chromia bricks.

Background: In recent years, the disease outbreak caused by Vibrio harveyi upset the booming development of the Chinese tongue sole (Cynoglossus semilaevis) farming industry. Genomic selection (GS) is a powerful method to improve the traits of interest, which has been proved in livestock and some fishes. Besides, the single nucleotide polymorphism (SNP) array is an efficient genotyping platform that can be used for genetic studies. To improve V. harveyi resistance in C. semilaevis, we firstly constructed a reference group of 1,572 individuals and investigated accuracies of four genomic methods (genomic best linear unbiased prediction (GBLUP), weighted GBLUP, BayesB, and BayesC) at predicting the genomic estimated breeding value (GEBV) using five-fold cross-validation and SNPs varying from 0.5 k to 500 k. Then, an SNP array was developed using the Affymetrix Axiom technology, and its accuracy in genotyping was evaluated by comparing SNPs generated by the array and by the re-sequencing technology. Finally, we selected 44 candidates as the parents of 23 families of C. semilaevis to evaluate the feasibility of the SNP array for GS.Results: all genomic methods outperformed the pedigree-based BLUP (ABLUP) when at least 50 k SNPs used for prediction, of which GBLUP resulted in better estimation than ABLUP when more than 1 k SNPs used. A 38 k SNP array, “Solechip No.1”, was developed with an average of 10.5 kb inter-spacing between two adjacent SNPs. The SNPs generated by the array and by the re-sequencing reached an average consistency of 94.8 %, of which 79.3 % of loci had a more than 90 % of the consistency. The survival rates of these 23 offspring families had a correlation of 0.706 with the family GEBVs (mid-parental GEBVs), and the average survival rate of the top five families in GEBVs (79.1 %) is higher than the bottom five families (58.1 %).Conclusion: GS is an efficient method to improve the V. harveyi resistance in C. semilaevis, and the SNP array “Solechip No.1” is a convenient and reliable tool for the Chinses tongue sole selective breeding practice.

The typical sexual size dimorphism (SSD) phenomenon of Chinese tongue sole (Cynoglossus semilaevis) seriously restricts the sustainable development of the fishing industry. Previous transcriptome analysis has found a close relationship between the steroid biosynthesis and C. semilaevis SSD. The 7-dehydrocholesterol reductase (dhcr7) and lathosterol 5-desaturase (sc5d) are two genes in the steroid biosynthesis pathway, playing important roles in lipid synthesis, cellular metabolism, and growth. The present study assessed their roles in the mechanism of C. semilaevis SSD. The quantitative polymerase chain reaction (qPCR) results showed that C. semilaevis dhcr7 was mainly expressed in female livers, and C. semilaevis sc5d was highly expressed in female livers and gonads. Dual-luciferase experiment showed that dhcr7 and sc5d promoters had strong transcriptional activity. The transcription factors E2F transcription factor 1 (E2F1), and CCAAT enhancer binding protein alpha (C/EBPα) significantly regulated the transcriptional activity of dhcr7 and sc5d promoters, respectively. Furthermore, small interfering RNA (siRNA) knockdown results showed that expression levels of several genes [SREBF chaperone (scap), membrane-bound transcription factor peptidase, site 1 (mbtps1), fatty acid synthase (fasn), sonic hedgehog (shh), bone morphogenetic protein 2b (bmp2b) and AKT serine/threonine kinase 1 (akt1)] were suppressed. Protein subcellular localization results indicated that Dhcr7 and Sc5d were both specifically distributed in the cytoplasm, with co-localization been observed. The present study provides evidence that dhcr7 and sc5d might regulate C. semilaevis sexual size dimorphism by involving in energy homeostasis and cell cycle, or by affecting PI3K-Akt and Shh signaling pathways. The detailed roles of these steroid biosynthesis genes regulating C. semilaevis SSD needed more information.

A polylactide composite fracture fixator loaded with vancomycin cationic liposome (PLA@VL) was prepared by reverse evaporation method. The method of cationic liposome encapsulating vancomycin could effectively improve antibacterial property and achieve drug sustained release effect, so as to reduce toxicity of antibiotics in vivo. Scanning electron microscope (SEM) was used to observe morphology and Fourier transform infrared spectroscopy (FTIR) was used to detect the composition of the internal fixator. In vitro drug release model, in vitro degradation model and body fluid osteogenesis model were designed in this study. On the other hand, the experiments of inhibition zone and MC3T3-E1 osteoblasts in mice were conducted to explore antibacterial property, cell activity and adhesion of the PLA@VL composite internal fixator. Alkaline phosphatase (ALP) staining method and alizarin red assay were used to detect the osteogenic induction ability of the composite internal fixator. Finally, mice fracture models were established to verify osteogenic and anti-infection abilities of the composite internal fixator in vivo. The results showed that MC3T3-E1 cells had better adhesion and proliferation abilities on the PLA@VL composite internal fixator than on the PLA fixator, which indicated that the PLA@VL composite internal fixator possessed excellent osteogenic and anti-infection abilities both in vivo and in vitro. Therefore, the above experiments showed that the fracture internal fixator combined with vancomycin cationic liposome had better biocompatibility, antibacterial ability and osteogenic ability, which provides a promising anti-infection material for the clinical field of fracture.

Corrosion resistance of four typical refractories against coal-water slurry coal slag, was investigated in laboratory. These typical refractories are high chromia brick (Cr2O3–Al2O3–ZrO2), fused AZS brick (Al2O3–ZrO2–SiO2), high zirconia brick (ZrO2) and Si3N4 bonded SiC brick, respectively. The cup test and micro-structural analysis method were used to evaluate the corrosion resistance and reaction mechanism at high temperature. Meanwhile, potential reactions between refractories and slag are predicted using thermodynamic calculation. The result shows that infiltrated depth increases in following order: Si3N4 bonded SiC brick

A systematic and generalized asymptotic derivative estimator design method is first presented for unknown maps by adding a sinusoidal excitation signal to the argument of the map. Then, based on the proposed asymptotic derivative estimator approach and based on the existing design methods for both finite-time and fixed-time state observers, finite-time and fixed-time derivative estimators are designed. The sufficient conditions for finite-time and fixed-time input-to-state stable of the finite-time and fixed-time derivative estimators are given respectively when a bounded disturbance input exists.

Alcohol intake increases the risk of cancer development. Approximately 3.6% human cancers worldwide derive from chronic alcohol drinking, including oral, liver, breast and other organs. Our studies in vivo and in vitro have demonstrated that diluted ethanol increase RNA Pol III gene transcription and promotes cell proliferation and transformation, as well as tumor formation. However, it is unclear about the effect of red wines on the human cancer cells. In present study, we investigated the roles of red wine in human cancer cell growth, colony formation and RNA Pol III gene transcription. Low concentration (12.5 mM to 25 mM) of ethanol enhances cell proliferation of breast and esophageal cancer lines, whereas its higher concentration (100 mM to 200 mM) slightly decreases the rates. In contrast, red wines significantly repress cell proliferation of different human cancer lines from low dose to high dose. The results reveal that the red wine also inhibits colony formation of human breast cancer and esophageal carcinoma cells. The effects of repression on different human cancer lines are in a dose-dependent manner. Further analysis indicates that ethanol increases RNA Pol III gene transcription, whereas the red wines significantly reduce transcription of the genes. Interestingly, the effects of mature wine (brick red) on cancer cell phenotypes are much stronger than young wine (intense violet). Together, these new findings suggest that red wines may contain some bioactive components, which are able to inhibit human cancer cell growth and colony formation.

Lhx9 is a LIM-homeodomain protein related to gonad development and sex reversal. In this study, we cloned and characterized CS-Lhx9 in the gonads of the Chinese tongue sole (Cynoglossus semilaevis). The full-length cDNA of CS-Lhx9 was 3123 bp, including an ORF of 1149 bp encoding 383 amino acids which contains two LIM domains and a homeobox domain. CS-Lhx9 transcripts were primarily observed in the testis of male and neomale at 1 yah, but nearly undetectable in the ovary. During the development of gonad, CS-Lhx9 exhibited an increasing trend and appeared to reach its peak value of expression in testis at 1 yah. In situ hybridization was performed in male and neomale gonads at 210 dah and 1 yah. The results showed strong CS-Lhx9 signals in the spermatids and spermatozoa (germ cells). Methylation level of CS-Lhx9 promoter was higher in female and lower in male and neomale gonads, showing a negative correlation with CS-Lhx9 expression.

The healthcare industry is confronted with the challenge of offering customized services while in the meantime to control increasing healthcare costs. Modularization is an important approach to reduce healthcare costs and improve patient-centered services via decreasing process complexity and enhancing flexibility through configuring pre-identified service modules. Recognizing the importance of modularity for healthcare services, this paper introduces Design Structure Matrix (DSM) as a technique for healthcare process modularization. A DSM-based modularization and sequencing algorithm is developed to allocate healthcare activities to service modules using Genetic Algorithm (GA) and arrange sequences of services both within and across service modules to support modular clinical pathway design. The proposed algorithm is implemented with a case study, the results of which have demonstrated the feasibility and applicability of the proposed DSM-based modularization method for healthcare process design. This research is partially supported by the National Natural Science Foundation of China under Project Number 71873015.

Rac1 and Rac2, belonging to the small Rho GTPase family, play an important role during the immune responses. In this study, a Rac1 homolog (CsRac1) and a Rac2 homolog (CsRac2) were cloned from the Cynoglossus semilaevis. The full-length of CsRac1 and CsRac2 cDNA was 1219 bp and 1047 bp, respectively. Both CsRac1 and CsRac2 contain a 579 bp open reading frame (ORF) which encoding a 192 amino acids putative protein. The predicted molecular weight of CsRac1 and CsRac2 was 21.41 kDa and 21.35 kDa, and their theoretical pI was 8.50 and 7.91, respectively. Sequence analysis showed that the conserved RHO domain was detected both from amino acid of CsRac1 and CsRac2. Homologous analysis showed that CsRac1 and CsRac2 share high conservation with other counterparts from different species. The CsRac1 and CsRac2 transcript showed wide tissue distribution, in which CsRac1 and CsRac2 exhibit the highest expression level in liver and gill, respectively. The expression level of CsRac1 and CsRac2 fluctuated in the liver and gill tissues at different time points after challenged by Vibrio harveyi. Specifically, CsRac1 and CsRac2 were significantly up-regulated at 48 h and 96 h post injection. Moreover, the knocking down of CsRac1 and CsRac2 in cell line (TSHKC) reduced the expression of CsPAK1, CsIL1-β and CsTNF-α. The present data suggests that CsRac1 and CsRac2 might play important roles in the innate immunity of half-smooth tongue sole.

Sexual size dimorphism (SSD), whereby females and males exhibit different body sizes, are widely documented in animals. To explore crucial regulators implicated in female-biased SSD of Chinese tongue sole (Cynoglossus semilaevis), GWAS was conducted on 350 females and 59 males. Twenty SNPs and 25 genes including zbed1, nsd3, cdc45, klhl29, and smad4 with -log(p) 7 were screened, mainly mapping to sex chromosome. The chromosome W-linked gene zbed1 attracted particular attention because it is a master key for cell proliferation. Thus, the regulatory network of zbed1 in C. semilaevis was explored by DAP-seq and 1352 peaks were discovered in the female brain. Moreover, zbed1 potentially regulated hippo signaling pathway, cell cycle, translation, and PI3k-Akt signaling pathway in C. semilaevis. These findings identify crucial SNPs and genes associated with female-biased SSD in C. semilaevis, also provide the first genome-wide survey for the zbed1 regulatory network in fish species.

In mammals, microtubule-dependent trafficking could participate the immune response, where the motor proteins are suggested to play an important role in this process, while the related study in fish was rare. In this study, dctn5, a subunit of dyactin complex for docking motor protein, was obtained by previous immune QTL screening. The full-length cDNAs of two dctn5 transcript variants were cloned and identified (named dctn5_tv1 and dctn5_tv2, respectively). Tissue distribution showed that dctn5_tv1 was widely distributed and high transcription was observed in immune tissue (skin), while dctn5_tv2 was predominantly detected in gonad and very low in other tissues. Time-course expression analysis revealed that dctn5_tv1 could be up-regulated in gill, intestine, skin, spleen, and kidney after Vibrio harveyi challenge. Moreover, recombinant Dctn5_tv1 exhibited high antimicrobial activity against Escherichia coli and Streptococcus agalactiae due to binding to bacteria cells. Taken together, these data suggest Dctn5_tv1 is involved in immune response of bacterial invasion in Chinese tongue sole.

Multi-camera vision systems (MVS) have a larger field of view than a single camera and are often applied to acquire geometric information of large-scale objects or scenes. Data integration from multiple cameras is a very important part for vision systems which is usually solved by using a global calibration method. After installing the MVS, in some cases, cameras do not have a common field of view (CFV) or only a narrow CFV. Accurate global calibration of cameras with non-overlapping field of view (FOV) is a very challenging task. A variety of global calibration of non-overlapping multi-camera methods (GCNM) have been proposed to estimate the relative positions and orientations of cameras based on different types of media or techniques such as large-range measuring devices, large-scale calibration targets, optical mirrors, motion model, laser projection, visual measuring instruments, etc. However, the GCNM is not yet a completely solved problem. Choosing which type of GCNM method to use is highly dependent on the specific vision system. Thus, in this paper, we present a comparative review of different GCNM methods and analyze accuracy, range, defects, and applications. Researchers and developers can take it as background information for their future works.

Despite Japanese flounder ( Paralichthys olivaceus ) is of importance in aquaculture of Northeast Asia, selective breeding programs for growth performance traits remain limited. The aim of this study was to estimate parameters of growth performance traits in this species. The heritabilities, genetic and phenotypic correlations, and breeding values were estimated for body weight (BW), total length (TL), condition factor (K), and average daily gain (ADG). The genetic analyses were performed on a total number of 186 full-sib families (108 sires and 146dams, 16,807 individuals with integrated pedigree information) from 4 year-classes based on three mixed animal models with tank and year-class as fixed effects, mean family trait values at tagging as a covariate, and additive genetic effect as a random effect using the restricted maximum likelihood method. For all growth traits the maternal and sir-dam interaction effects in proportion to phenotypic variance were very low (0.00–0.05). The heritability estimates for growth traits ranged from 0.12 to 0.39, which is in a low to moderate level. These results indicated that there is a considerable additive genetic variation in growth traits, and the ongoing selective breeding project will produce considerable genetic improvement in growth traits of Japanese flounder. The genetic and phenotypic correlations among BW, TL and ADG were high and positive (0.87–0.94, P Statement of relevance This paper offers guidelines to select breeding strategy in Japanese flounder.

In mammals, Class II, major histocompatibility complex (MHC II) transactivator (CIITA) recognizes microbial pathogens and triggers immune responses. In Chinese tongue sole Cynoglossus semilaevis, Cs-CIITA was prevalently expressed in various tissues. Cs-CIITA, Cs-MHC IIA and Cs-MHC IIB were expressed significantly higher in skin in susceptible families infected with Vibrio harveyi, while higher expression of Cs-CIITA and Cs-MHC IIB was examined in liver in resistant families. In addition, the three genes were up-regulated in gill, skin, intestine, liver, spleen and kidney at 48 h or 72 h after V. harveyi infection. Furthermore, the three genes were co-expressed in the epithelial mucous cells of gill, skin, and intestine. Knockdown of Cs-CIITA regulates the expression of other inflammation-related genes, including CD40, IL-1β, IL-6, RelB, cRel, NFκB, and Myd88. These results suggest that CIITA functions in the inflammatory responses of C. semilaevis against V. harveyi, via MHC II transcriptional regulation.

In this paper, we consider a coordinated multi-aircraft 4D (3D space plus time) trajectories planning problem which is illustrated by planning 4D trajectories for aircraft traversing an Air Traffic Control (ATC) sector. The planned 4D trajectories need to specify each aircraft’s position at any time, ensuring conflict-free and reducing fuel and delay costs, with possible aircraft maneuvers such as speed adjustment and flight level change. Different from most existing literature, the impact of buffer safety distance is also under consideration, and conflict-free is guaranteed at any given time (not only at discrete time instances). The problem is formulated as a pure-strategy game with aircraft as players and all possible 4D trajectories as strategies. An efficient maximum improvement distributed algorithm is developed to find equilibrium at which every aircraft cannot unilaterally improve further, without enumerating all possible 4D trajectories in advance. Proof of existence of the equilibrium and convergence of the algorithm are given. A case study based on real air traffic data shows that the algorithm is able to solve 4D trajectories for online application with estimated 16.7% reduction in monetary costs, and allocate abundant buffer safety distance at minimum separation point. Scalability of the algorithm is verified by computational experiments.

Alcohol intake is associated with numbers of different human cancers, such as hepatocellular carcinoma (HCC) and breast cancer. However, the molecular mechanism remains to be elucidated. RNA polymerase III-dependent genes (Pol III genes) deregulation elevates cellular production of tRNAs and 5S rRNA, resulting in an increase in translational capacity, which promote cell transformation and tumor formation. To explore a common mechanism of alcohol-associated human cancers, we have comparably analyzed that alcohol causes deregulation of Pol III genes in liver and breast cells. Our results reveal that alcohol enhances RNA Pol III gene transcription in both liver and breast cells. The induction of Pol III genes caused by alcohol in ER+ breast cancer lines or liver tumor lines are significantly higher than in their non-tumor cell lines. Alcohol increases cellular levels of Brf1 mRNA and protein, (which depeted) Brf1 is a key transcription factor and specifically regulate Pol III gene activity. Alcohol activates JNK1 to upregulate transcription of Brf1 and Pol III genes, whereas inhibition of JNK1 by SP600125 or its siRNA significantly decreases the induction of these genes. Furthermore, alcohol increases the rates of transformation of liver and breast cells, repressed JNK1 and Brf1 expression decrease transcription of Pol III genes and reduce the rates of colony formation of AML-12 and MCF-10 cells. Together, these studies support the idea that alcohol induces deregulation of Brf1 and RNA Pol III genes in liver and breast cells, which share a common signaling pathway to promote cell transformation. Through the common mechanism, alcohol-induced deregulation of RNA Pol III genes brings about greater phenotypic changes.

A hybrid method with high reliability to detect ellipses in low SNR industrial images is proposed.Using candidate elliptic arcs with high fitting precision as seed arcs to group with other arcs.A novel method to extract regions which may contain one ellipse is proposed.New selection scores for evaluating hypothetical ellipse are used.Experimental results show high reliability of proposed method. Many ellipse detection methods have been proposed for detecting ellipses in images. However, they are unsuitable for industrial images due to low signal-to-noise ratios (SNR). This paper presents an ellipse detection method combining the advantages of Hough transform(HT) based methods and the advantages of edge following methods, which is capable of detecting fragmented ellipses and is both computational and memory efficient. Our method works in two steps. In the first step, an edge following method is proposed to quickly and accurately extract the majority of ellipses. For ellipses missed in the first step, candidate regions where each may contain one missed ellipse are extracted in the second step using cluster analysis, and then a HT based method is performed on these regions to extract the missed ellipses. This can not only guarantee the accuracy of the HT based method, but also save the memory and computation time. We test the performance of our method using both synthetic images and low SNR industrial images. Experimental results demonstrate that the proposed method performs far better than existing methods in terms of recall, precision, F-measure, and reliability. Especially in term of reliability, our method has achieved a very high value close to 1 while the reliabilities of state-of-the art methods are almost less than 0.5.

In previous research, the full-length sghC1q (CssghC1q) recombinant protein from half-smooth tongue sole (Cynoglossus semilaevis) displayed antibacterial and antiviral activities. However, the roles of the different domains (gC1q and coiled-coil domains) in CssghC1q are still unknown. Thus, in this study, with the help of the pET-28a vector, recombinant proteins comprising different CssghC1q domains were constructed and verified. However, only the recombinant pET-28a-CsgC1q protein was successfully purified. The pET-28a-CsgC1q recombinant protein only exhibited antibacterial activities against Vibrio anguillarum among the bacteria tested. Compared with the full-domain recombinant protein, the recombinant CsgC1q domain exhibited a very low antibacterial activity (10%) at very high protein concentrations (0.1 mg/ml). The CsgC1q protein could show 80% antimicrobial activity against V. anguillarum, though a very high concentration (2.25 mg/ml) was necessary. The CsgC1q protein, which displayed the antibacterial activities, could require its coiled-coil region to promote correct protein folding; thus, without this domain, the protein would fold inefficiently. The results indicate that the CsgC1q domain may be the key antibacterial activities function domain. However, the CsgC1q domain alone is insufficient for proper antibacterial activity. The coiled-coil region, which is located upstream of the CsgC1q domain, might be important for proper folding and antibacterial activities.

In the present work, we investigate a repairable service parts inventory system that has a central repair facility and several locations storing inventory called bases. If a part fails, then the failed part is identified and replaced with a ready-to-use part from the base. Afterwards, the failed part is sent to the repair facility, where it is repaired and allocated to one of the bases, with the objective being to identify the base with the most urgent need of a service part to minimize the expected backorder cost. To achieve this, we examine the initial base-stock provisioning problem in conjunction with real time stock allocation decision making. By modeling the problem as a Markov decision process, we characterize the optimal solution for each decision and prove that identifying the optimal policy for one of the decisions leads to the optimal solution for the other. Considering the computational intensity of the multi-base problem, we propose a heuristic technique for the stock allocation problem based on relative value function and average backorder cost at a single base. Further, we compare the performance of the heuristic model with the myopic policy, which is widely applied in the literature, to validate the efficiency of our proposed heuristic. A sensitivity analysis is carried out to illustrate the effects of important problem parameters on the performance of the presented heuristic. Results reveal that the proposed stock allocation policy outperforms the myopic policy.

In this paper, we demonstrate a plasmon-induced transparency (PIT) sample based on a hybrid structure of a metasurface/BiFeO3/Si, which is theoretically and experimentally investigated in the terahertz range. A distinct transparency window can be available in the transmission spectra, which originates from the near-field coupling of two resonators. A modulation depth of 82% can be obtained at the transparency window through tuning the optical pumping power. The photo-excited carriers suppress the resonances of the two bright modes, then the transparency window of the PIT metasurface is nearly disappeared. This work provides a new perspective on exploring the multiferroic material BiFeO3 as well as expanding the practical applications of BFO-based functional devices in the terahertz range.

Thermal stress, encountered frequently in aquaculture, affects diverse physiological processes and behavior of fish; however, the mechanism underlying these effects may vary among species. Half-smooth tongue sole (Cynoglossus semilaevis) widely inhabits Asian coastal waters and has been intensively cultured in China. In this study, the transcriptomes of three tissues (gill, liver and muscle) of half-smooth tongue sole stressed at temperatures up to 35°C were profiled and compared with those of the fish living at a normal temperature of 25°C. Of 105,228 transcripts, 23,213 were annotated in GO terms, and 718, 1236 and 561 were found to express differentially in gill, liver and muscle from control, respectively. Of the differentially expressed genes, 119 were shared by all three tissues. Tissue-overlapping transcripts and the pathways and functions they defined may underline the primary response of fish to thermostressing. High temperature may cause directly protein mis-folding and hypoxia. Cells suffering thermostress either survive hypoxia or are eliminated by immune system. High temperature may have interacted indirectly through HSPs with HIF-1 and JAKs/STATs signaling pathways. The former up-regulates the expression of hypoxia inducible genes while the later up-regulates the expression of genes associating with insulin and NOD-like receptors signaling pathways. Insulin pathway functions to sustain homeostasis of sugar and lipids, aiding to survive cells, while NOD-like receptors signaling pathway strengthens immune, apoptotic and inflammatory responses, helping to survive cells. These understandings may help improve our culturing practice, the culture performance of half-smooth tongue sole and its breeding.

To enhance the dynamic and static performance of the extremum seeking control (ESC) scheme, a novel fast ESC scheme without steady-state oscillation is proposed, in which the structure of the classic ESC scheme is adjusted to make the sinusoidal excitation signal amplitude locally exponentially converge to zero. The improved ESC scheme can speed up the convergence to shorten the seeking time dramatically, and enlarge the search area to avoid falling to local extrema effectively. It can also reduce the sinusoidal excitation signal amplitude to eliminate the adverse effects of the steady-state oscillation eventually. The rigorous stability analysis and proof of the improved ESC scheme are provided in detail, and the simulation results are presented to illustrate its effectiveness and superiority. Finally, the application of the improved ESC scheme to antilock braking systems (ABS) has been discussed through comparing with classical perturbation-based ESC scheme and sliding-mode-based ESC scheme to illustrate the practicability of the improved scheme.

CCR6 is an important binding receptor of CCL20 and beta-defensins, and has multiple functions in the innate and acquired immune responses. In this study, we cloned the PoCCR6A and PoCCR6B genes of the Japanese flounder and studied the gene structure and expression patterns of these two genes in bacterial infection. The full-length PoCCR6A cDNA is 1415 bp and the open reading frame (ORF) is 1113 bp, encoding a 370-amino-acid peptide. The full-length PoCCR6B cDNA is 2193 bp and the ORF is 1029 bp, encoding a 363-amino-acid peptide. The structures of PoCCR6A and PoCCR6B indicate that they are single-exon genes. The predicted proteins encoded by PoCCR6A and PoCCR6B have the typical G-protein-coupled receptor (GPCR) family signature of seven transmembrane domains and several conserved structural features. A tissue distribution analysis showed that PoCCR6A is predominately expressed in the intestine, gill, and blood, and PoCCR6B in the gill, spleen, and liver. The expression patterns of the two chemokine receptors were analyzed during bacterial infection. In spleen and kidney, the expression of PoCCR6A was significantly upregulated at 24 h after infection, whereas the expression of PoCCR6B was steady at these time points. While in intestine, both of them were upregulated at 6 h-12 h after infection, and in gill the expression levels of them were upregulated at 24 h. The patterns of expression suggested that PoCCR6A and PoCCR6B play an important role in the immune response of the Japanese flounder, especially in the mucosal tissues.

Complement factor I (Cfi) is a soluble serine protease which plays a crucial role in the modulation of complement cascades. In the presence of substrate modulating cofactors (such as complement factor H, C4bp, CR1, etc), Cfi cleaves and inactivates C3b and C4b, thereby controlling the complement-mediated processes. In this study, we sequenced and characterized Cfi gene from Cynoglossus Semilaevis (designated as CsCfi) for the first time. The full-length cDNA of CsCfi was 2230 bp in length, including a 98 bp 5'-untranslated region (UTR), a 164 bp 3'-UTR and a 1968 bp open reading frame (ORF). It encoded a polypeptide of 656 amino acids, with a molecular mass of 72.28 kDa and an isoelectric point of 7.71. A signal peptide was defined at N-terminus, resulting in a 626-residue mature protein. Multiple sequence alignment revealed that Cfi proteins were well conserved with the typical modular architecture and identical active sites throughout the vertebrates, which suggested the conserved function of Cfi. Phylogenetic analysis indicated that CsCfi and the homologous Cfi sequences from teleosts clustered into a clade, separating from another clade from the cartilaginous fish and other vertebrates. Tissue expression profile analysis by quantitative real-time PCR (qRT-PCR) showed that CsCfi mRNA constitutively expressed in all tested tissues, with the predominant expression in liver and the lowest in stomach. Temporal expression levels of CsCfi after challenging with Vibrio anguillarum showed different expression patterns in intestine, spleen, skin, blood, head kidney and liver. The recombinant CsCfi (rCsCfi) protein showed broad-spectrum antimicrobial activities against the Gram-positive bacteria Staphylococcus aureus and the Gram-negative bacteria Escherichia coli, Pseudomonas aeruginosa and Shewanella putrefaciens. The research revealed that CsCfi plays an important role in C. Semilaevis immunity.

China is the largest producer of tilapia in the world, and genetically improved farmed tilapia (GIFT) strain of Oreochromis niloticus is the main breed which is cultured widely in coastal and inland areas of China. Streptococcal disease, caused by Streptococcus agalactiae, is one of the most prevalent diseases in farmed tilapia that causes huge economic loss, leading to urgent needs for disease-resistant breed of tilapia. In view of the predictive accuracy and selective efficiency, genomic selection (GS) could be a feasible and optimal method to improve the S. agalactiae resistance in GITF. In this study, the genetic architecture of resistance to S. agalactiae and the accuracy of three GS methods at improving the resistance in GIFT were investigated by artificial challenge test (for phenotypes) and whole-genome re-sequencing (for genotypes). A total of 897 individuals were genotyped (776 individuals were used as the reference group) and 3,759,557 high-quality SNPs were obtained for downstream analysis. The genome-wide association study (GWAS) using weighted genomic BLUP (wGBLUP) and 1 M SNPs revealed that resistance to S. agalactiae in GIFT belongs to a polygenic genetic architecture. Seven suggestive trait-related SNPs were identified on chromosomes 4, 7, 11, and 18. These SNPs explained more than 1% genetic variance. Three GS (GBLUP, wGBLUP, and BayesCπ) methods had better estimation than pedigree-based BLUP, which indicated that the genomic selection approach could be a feasible and promising method to improve the S. agalactiae resistance in GIFT. Although wGBLUP and BayesCπ resulted in similar estimation accuracy, wGBLUP had priority when estimating time needs to be considered.

Edwardsiella tarda is one of the most prevalent and economically important diseases worldwide in aquaculture, including olive flounder (Paralichthys olivaceus). Genetic improvement for resistance to E. tarda is an alternative strategy for the prevention of disease outbreaks. The objective of this work was to evaluate the genetic parameters of disease resistance by challenge test data. Mortality was recorded daily for 16 days (day1 to 16). At day 4 and 16, the cumulative mortality was 52%, and 87%, respectively. Resistance was defined as binary test survival (BTS), day of death (DD), and binary test day survival (BTDS). For DD, only day 16 was set as the cut-off day, while for the other two traits, each test day was set as a cut-off day. These traits were fitted in four statistical models for estimating variance components and calculating genetic parameters. Namely, 1) cross-sectional linear model for BTS (CLM) and DD (LM), 2) threshold (logit) model (CTM) for BTS, and 3) linear (logit) repeatability model (LRM) for BTDS. In LM, the heritability of resistance was 0.23 ± 0.06. In other three models, heritabilities were estimated at each cut-off point (test day) respectively, and heritabilites fluctuated daily. At day 4 (with approximately 50% overall mortality), the heritability was 0.17 ± 0.11 for CLM, 0.20 ± 0.14 for CTM and 0.09 ± 0.08 for LRM. Heritabilities obtained based on LM (at day16) and other three model estimated at day 4 are considered to be accurate. The selection accuracies of LM and LRM are nearly identical (0.735 and 0.755 respectively) and are 9.5% to13.9% higher than values of CLM and CTM (0.663 and 0.671 respectively). The rank correlations between EBVs obtained from CLM, CTM and LRM were very high (ranging from 0.97–1.00), indicating a near identical ranking of families. However, the correlations between EBVs from LM and from other three models were substantially lower (ranging from 0.79–0.82), which imply a substantial reranking of families. Low to moderate heritabilities indicated that resistance against E. tarda can be genetically improved, and selection by using models (i.e. LM and LRM) which take time to death into account is more accurate than cross-sectional models.

Configuration has been recognized as an effective methodology to provide high product variety that caters to individual customer’s needs in the manufacturing industry. For healthcare service configuration, this decision-making process can be taken as service package creation and treated analogously as a project, which is defined as a collection of tasks. This research develops a decision support model to integrate individual patients into the healthcare service configuration process. The healthcare service configuration is formulated as a Resource Constrained Project Scheduling Problem (RCPSP), and a bi-level optimization algorithm based on Genetic Algorithm (GA) is developed for problem solving. The methodology and algorithm are implemented with a case study based on the data obtained from a general hospital in Singapore, which has demonstrated the applicability of healthcare service configuration.

Half-smooth tongue sole (Cynoglossus semilaevis) is one of the most valuable marine aquatic species in Northern China. Given to the rapid development of aquaculture industry, the C. semilaevis was subjected to disease-causing bacteria Vibrio anguillarum. It therefore is indispensable and urgent to understand the mechanism of C. semilaevis host defense against V. anguillarum infection. In the present study, the extensively analysis at the transcriptome level for V. Anguillarum disease in tongue sole was carried out. In total, 94,716 high quality contigs were generated from 75,884,572 clean reads in three libraries (HOSG, NOSG, and CG). 22,746 unigenes were identified when compared with SwissProt, an NR protein database and NT nucleotide database. 954 genes exhibiting the differentially expression at least one pair of comparison in all three libraries were identified. GO enrichment for these genes revealed gene response to biotic stimulus, immune system regulation, and immune response and cytokine production. Further, the pathways such as complement and coagulation cascades and Vibrio cholerae infection pathways were enriched in defensing of pathogen. Besides, 13,428 SSRs and 118,239 SNPs were detected in tongue sole, providing further support for genetic variation and marker-assisted selection in future. In summary, this study identifies several putative immune pathways and candidate genes deserving further investigation in the context of development of therapeutic regimens and lays the foundation for selecting resistant lines of C. semilaevis against V. anguillarum.

The C1q family includes many proteins that contain a globular (gC1q) domain, and this family is widely conserved from bacteria to mammals. The family is divided into three subgroups: C1q, C1q-like and ghC1q. In this study, a novel C1q family member, s ghC1q , was cloned and identified from Cynoglossus semilaevis (named CssghC1q ). The full-length CssghC1q cDNA spans 905 bp, including an open reading frame (ORF) of 768 bp, a 5′-untranslated region (UTR) of 25 bp and a 3′-UTR of 112 bp. The ORF encodes a putative protein of 255 amino acids (aa) with a deduced molecular weight of 28 kDa. The predicted protein contains a signal peptide (aa 1–19), a coiled-coil region (aa 61–102) and a globular C1q (gC1q) domain (aa 117–255). Protein sequence alignment indicated that the C-terminus of CssghC1q is highly conserved across several species. Phylogenetic analysis indicated that CssghC1q is most closely related to Maylandia zebra C1q-like-2-like . The CssghC1q genomic sequence spanned 1562 bp, with three exons and two introns. CssghC1q is constitutively expressed in all evaluated tissues, with the highest expression in the liver and the weakest in the heart. After a challenge with Vibrio anguillarum, CssghC1q transcript levels exhibited distinct time-dependent response patterns in the blood, head kidney, skin, spleen, intestine and liver. Recombinant CssghC1q protein exhibited antimicrobial activities against Gram-negative bacteria, Gram-positive bacteria and viruses. The minimum inhibitory concentration (MIC) values against Vibrio harveyi, Vibrio anguillarum, Pseudomonas aeruginosa and Staphylococcus aureus were 0.043 mg/mL, 0.087 mg/mL, 0.174 mg/mL and 0.025 mg/mL, respectively. A low concentration (0.06 mg/mL) of CssghC1q showed significant antiviral activity in vitro against nervous necrosis virus (NNV). These results suggest that CssghC1q plays a vital role in immune defense against bacteria and viruses.

article i nfo Article history: piwi is an important regulator gene in germ cell division during spermatogenesis. piwi homologous genes are in- volved in gametogenesis and germline specification, and knocking down these genes could affect germ cell mei- otic progression. To understand the function of piwi-related genes in spermatogenesis, we cloned a Piwi- subfamily member (piwil2 gene) from the gonad of Cynoglossus semilaevis. The full-length of piwil2 cDNA was 3314 bp, including a 3162 bp open reading frame (ORF), a 60 bp 5'-UTR, along with a 92 bp 3'-UTR, and encoded a predicted protein of 1053 amino acid residues. Phylogenetic analysis showed that the PIWIL2 putative protein belonged to the Argonaute protein family, and Piwi-subfamily, with typical PAZ and Piwi domains. Ultrathinsec- tionsof differentgonadalstages, and real-timequantitative PCR showed that therelative expression of thepiwil2 gene couldn't be detected until day 95 (95 days) larvae, when germ cell divided rapidly in C. semilaevis. The piwil2 transcripts were more abundant in gonads of males and neo-males than in females, and weakly expressed in other tissues and organs. Compared with the relative expression of piwil2 in gonads, the CpG methylation levels were significantly higher in females. Chromosomal fluorescence in situ hybridization (FISH) showed that the piwil2 gene was located on the Z sex chromosome of C. semilaevis. These results suggest that piwil2 plays an important role in spermatogenesis of C. semilaevis.

The growth arrest and DNA-damage-inducible protein 45 gamma (Gadd45g) is known to play a major role in embryonic development and sex determination. In this study, two Gadd45g genes were isolated from half-smooth tongue sole (Cynoglossus semilaevis). Using chromosomal fluorescence in situ hybridization (FISH), Gadd45g1 and Gadd45g2 were located on the W and Z chromosomes, respectively. The full-length cDNA sequences of Gadd45g1 (1270bp) and Gadd45g2 (1181bp) were predicted to contain a 480-bp coding sequence that could encode a protein of 159 amino acids residues. A phylogenetic tree showed that the predicted Gadd45g1 and Gadd45g2 amino acid sequences clustered closely in one branch. It is proposed that Gadd45g1 and Gadd45g2 are paralogous genes derived from the divergence of the sex chromosome. Ka/Ks ratios indicated that Gadd45g1 and Gadd45g2 may have undergone a high number of mutations and have a divergence time of only about 68,000years, although Gadd45g homologs are highly conserved. The qRT-PCR demonstrated that Gadd45g1 and Gadd45g2 were highly expressed in ovary, and negligibly expressed in testis of male and neo-male. During development of the ovary (from 80 to 150days), the expression levels of both genes reached high levels. Gadd45g1 was also highly expressed at 50days, the stage just before gonad differentiation in C. semilaevis. All these findings imply functional divergence of the two Gadd45g homologs; Gadd45g1 may be necessary for sex differentiation in the early stage of gonad development, and then Gadd45g1 and Gadd45g2 maintain ovary development and the female character of half-smooth tongue sole.