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Duck Tembusu virus (DTMUV) is a newly emerging pathogenic flavivirus that has caused huge economic losses to the duck industry in China since 2010. Moreover, the infection has spread rapidly, resulted in a potential public health concern. To improve our understanding of the host cellular responses to virus infection and the pathogenesis of DTMUV infection, we used RNA-Seq to detect the gene changes in DF-1 cells infected and mock-infected with DTMUV. A total of 663 differentially-expressed genes (DEGs) were identified in DTMUV-infected compared with mock-infected DF-1 cells at 24 h post-infection (hpi), among which 590 were up regulated and 73 were down regulated. Gene Ontology analysis indicated that the DEGs were mainly involved in cellular process, immune system processes, metabolic processes, and signal-organism process. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the DEGs were mainly involved in several signaling pathways such as Toll-like receptor signaling, Jak-STAT signaling, RIG-I-like receptor signaling and AGE-RAGE signaling pathway. Moreover, some selected DEGs were further confirmed by real-time PCR and the results were consistent with the sequencing data. To our knowledge, this study is the first to analyze the transcriptomic change in DF-1 cells following DTMUV infection. We believe that our research provides useful information in better understanding the host response to DTMUV infection and the inherent mechanism of DTMUV replication and pathogenicity.

Kinetic and affinity analysis of protein interactions reveals information on their related activities in biological processes. Herein, we established a system for evaluating the kinetics and affinity of the interaction between protein A and various IgG species on the surface of silica spheres of silica colloidal crystal (SCC) films by the extraordinary optical interference capabilities of 190 nm silica spheres after self-assembly. The equilibrium association constant (KA) was calculated by the equilibrium Langmuir model and nonlinear least-squares analysis of time-dependent data. The relative protein A/IgG binding affinity is human rabbitcowgoat. In addition, the competitive interaction of distinct species of IgG with protein A at the interface of SCC films was studied and performed. These findings may help with the use of protein A and other recognition components in a number of sensor types. Furthermore, this research might offer a novel approach to determining the kinetics and affinity of proteins on the surface of spheres particles, which may contribute to the development of the application of spheres particles in pharmaceutical science, biomedical engineering, and other techniques.

Avian influenza virus (AIV) subtype H9N2 is the most widespread AIV in poultry worldwide, causing great economic losses in the global poultry industry. Chickens and ducks are the major hosts and play essential roles in the transmission and evolution of H9N2 AIV. Vaccines are considered an effective strategy for fighting H9N2 infection. However, due to the differences in immune responses to infection, vaccines against H9N2 AIV suitable for use in both chickens and ducks have not been well studied. This study developed an inactivated H9N2 vaccine based on a duck-origin H9N2 AIV and assessed its effectiveness in the laboratory. The results showed that the inactivated H9N2 vaccine elicited significant haemagglutination inhibition (HI) antibodies in both chickens and ducks. Virus challenge experiments revealed that immunization with this vaccine significantly blocked virus shedding after infection by both homogenous and heterologous H9N2 viruses. The vaccine was efficacious in chicken and duck flocks under normal field conditions. We also found that egg-yolk antibodies were produced by laying birds immunized with the inactivated vaccine, and high levels of maternal antibodies were detected in the serum of the offspring. Taken together, our study showed that this inactivated H9N2 vaccine could be extremely favourable for the prevention of H9N2 in both chickens and ducks.

Tembusu virus (TMUV) is an avian-origined flavivirus that is prevalent in ducks and geese. TMUV causes reduced egg production and neurological problems, resulting in profound economic losses to the waterfowl industry. In the viral life cycle, cellular factors are required for viral entry, replication, assembly, release and so on. Heat shock protein 70 (HSP70) is reported to be involved in the replication of multiple viruses. In this study, we explored the roles of HSP70 in the TMUV life cycle. The results showed that TMUV infection induced HSP70 expression starting 12 h post-infection. An HSP70 inhibitor reduced TMUV viral RNA production and the number of virus particles, whereas an HSP70 activator enhanced the amount of viral RNA and virions that released from the cells. Further analysis revealed that HSP70 played important roles in the postentry stages of the TMUV life cycle, including viral replication, assembly and release. We also found that inhibition of HSP70 expression significantly reduced TMUV-induced apoptosis. Additionally, incubation of TMUV particles with an anti-HSP70 antibody significantly reduced viral infectivity, suggesting an association between HSP70 and TMUV particles. These results implicate HSP70 in the life cycle of TMUV, and therefore, targeting HSP70 may be a strategy for developing an anti-TMUV therapy.

Anthocyanins can help plants adapt and resist adverse environments and have important nutritional and medicinal effects on human beings. However, how environmental factors affect the anthocyanins accumulation of plants and how to improve the anthocyanins content of plants in different soils needs further exploration. Hence, this study aimed to investigate the effects of environmental factors on the accumulation of cyanidin, petunidin, malvidin, and delphinidin in the fruits of Lycium ruthenicum in sandy desert grassland (SS), gravel desert grassland (GD), and saline-alkali desert grassland (SD) in the lower reaches of the Shiyang River Basin. The variable importance screened the key environmental factors affecting anthocyanin accumulation in projection (VIP) and multiple stepwise regressions. The structural equation model (SEM) was established to understand how the climate and soil factors affect the total anthocyanin accumulation. For establishing soil nutrient optimization schemes by partial least squares regression (PLS) and the simplex algorithm used to improve the anthocyanin content in different types of desert grassland. In SS, electrical conductivity (EC) and microbial biomass carbon (SMBC) showed highly significant and positive effects on the content of total anthocyanin, cyanidin, and petunidin. In GD, soil moisture and microbial biomass nitrogen (SNBN) significantly negatively affected total anthocyanin content. In SD, catalase (CAT), phosphatase (PHO), and total potassium (TK) had the greatest impact on total anthocyanin content. It is indicated that the targeted improvement measures are necessary to increase anthocyanin content in the fruit of Lycium ruthenicum.

Duck Tembusu virus (DTMUV) is a newly emerging pathogenic flavivirus that has caused significant economic losses to the duck industry in China since 2010 due to egg production losses and neurological dysfunction. DTMUV is a public health concern because the infection spreads rapidly among birds. Retinoic acid-inducible gene-I (RIG-I)serves as an innate immune sensor and plays a key role in host antiviral defenses. Tripartite motif-containing protein 25 (TRIM25), an E3 ubiquitin ligase, is pivotal for RIG-I ubiquitination and activation. In addition, TRIM25 acts as an interferon-stimulated gene and mediates the antiviral activity. However, the effect of duck TRIM25 on DTMUV has not been assessed. Herein, we reportthe antiviral function of TRIM25 against DTMUV. First, we constructed the pcDNA3.1-c-myc-duTRIM25 plasmid. TRIM25 has a 2052 bp open reading frame that encodes a predicted 684 amino acid protein consisting of a RING finger domain, a B-box domain, a coiled-coil domain, and a PRY/SPRY domain. The protein sequence identity with chicken, mouse, and human TRIM25 is 69.7, 47.8, and 48.3%, respectively. TRIM25 was upregulated in BHK-21 cells, duck embryo fibroblasts, and 293T cellsupon DTMUV infection. The expression of viral RNA and proteins was significantly lower in cells over expressing TRIM25 than in control cells. Furthermore, siRNA-mediated silencing of TRIM25 increased the production of viral progeny. These results help elucidate the molecular mechanisms underlying the host response to DTMUV infection and suggest potential control measures for DTMUV outbreaks.

Relationships between biodiversity and multiple ecosystem functions (that is, ecosystem multifunctionality) are context-dependent. Both plant and soil microbial diversity have been reported to regulate ecosystem multifunctionality, but how their relative importance varies along environmental gradients remains poorly understood. Here, we relate plant and microbial diversity to soil multifunctionality across 130 dryland sites along a 4,000 km aridity gradient in northern China. Our results show a strong positive association between plant species richness and soil multifunctionality in less arid regions, whereas microbial diversity, in particular of fungi, is positively associated with multifunctionality in more arid regions. This shift in the relationships between plant or microbial diversity and soil multifunctionality occur at an aridity level of ∼0.8, the boundary between semiarid and arid climates, which is predicted to advance geographically ∼28% by the end of the current century. Our study highlights that biodiversity loss of plants and soil microorganisms may have especially strong consequences under low and high aridity conditions, respectively, which calls for climate-specific biodiversity conservation strategies to mitigate the effects of aridification.

Given the fact that excessive levels of reactive oxygen species (ROS) induce damage to proteins, lipids, and DNA, various ROS-generating agents and strategies have been explored to induce cell death and tumor destruction by generating ROS above toxic threshold. Unfortunately, hypoxia in tumor microenvironment (TME) not only promotes tumor metastasis but also enhances tumor resistance to the ROS-generated cancer therapies, thus leading to ineffective therapeutic outcomes. A variety of nanotechnology-based approaches that generate or release O2 continuously to overcome hypoxia in TME have showed promising results to improve the efficacy of ROS-generated cancer therapy. In this minireview, we present an overview of current nanomaterial-based strategies for advanced cancer therapy by modulating the hypoxia in the TME and promoting ROS generation. Particular emphasis is put on the O2 supply capability and mechanism of these nanoplatforms. Future challenges and opportunities of design consideration are also discussed. We believe that this review may provide some useful inspiration for the design and construction of other advanced nanomaterials with O2 supply ability for overcoming the tumor hypoxia-associated resistance of ROS-mediated cancer therapy and thus promoting ROS-generated cancer therapeutics.

An increasing number of studies have revealed that histone deacetylase (HDAC) mediated histone deacetylation is important for mammalian oocyte development. However, nonselective HDAC inhibitors (HDACi) were applied in most studies; the precise functions of specific HDAC classes during meiosis are poorly defined. In this study, the class IIa-specific HDACi MC1568 was used to reveal a crucial role of class IIa HDACs in the regulation of histone deacetylation during porcine oocyte meiosis. Besides, the functions of HDACs and histone acetyltransferases in regulating the balance of histone acetylation/deacetylation were also confirmed during oocyte maturation. After the validation of nontoxicity of MC1568 in maturation rate, spindle morphology, and chromosome alignment, effects of MC1568 on developmental competence of porcine somatic cell nuclear transfer (SCNT) embryos were evaluated, and data indicated that treatment with 10 μM MC1568 for 12 hours following electrical activation significantly enhanced the blastocyst rate and cell numbers. Moreover, results showed that optimal MC1568 treatment increased the H4K12 acetylation level in SCNT one cells and two cells. In addition, MC1568 treatment stimulated expression of the development-related genes OCT4, CDX2, SOX2, and NANOG in SCNT blastocysts. Collectively, our investigation uncovered a critical role of class IIa HDACs in the regulation of histone deacetylation during oocyte meiosis. Furthermore, for the first time, we showed that MC1568 can improve the in vitro development of porcine SCNT embryos. These findings provide an alternative HDACi for improving animal cloning efficiency and may shed more light on nuclear reprogramming.

Tembusu virus (TMUV) is a newly emerging flavivirus and has caused significant economic loss to the poultry industry in China. To date, the entry of TMUV into host cells remains poorly understood. Here, the mechanism of TMUV entry into BHK-21 cells was investigated. The depletion of cellular cholesterol by methyl-β-cyclodextrin led to a significant decline in the titers and RNA levels of the infectious TMUV. This reduction was restored by supplementation of exogenous cholesterol. Membrane cholesterol depletion mainly blocked viral internalization but not attachment. However, viral infection was unaffected by genistein treatment or caveolin-1 silencing by small interfering RNA. In addition, clathrin-mediated endocytosis might be utilized in TMUV entry given that the viral infection was inhibited by knockdown of clathrin heavy chain and treatment of chlorpromazine (CPZ). Moreover, the number of internalized virus particles decreased under CPZ treatment. Dynasore inhibited TMUV entry suggesting a role for dynamin. Our results reveal that TMUV entry into BHK-21 cells is dependent on cholesterol, clathrin and dynamin but not caveolae.

Avian influenza subtype H9N2 infection is a mild but highly contagious disease that is associated with a decrease in the efficacy of vaccine interventions, and an increase in susceptibility to secondary infections in poultry. However, the immune evasion mechanism of H9N2 avian influenza viruses (AIVs) in chickens is poorly understood. Dendritic cells (DCs) are immune cells of major importance, involved in innate immune responses against viruses, but also in the setting of adaptive immune response due to their high ability to present viral antigen. Therefore, in the present study we used high-throughput RNA-sequencing technology at the transcriptome level to identify the differentially expressed genes (DEGs) between chicken DCs infected with H9N2 virus and mock-infected DCs. We identified 4151 upregulated DEGs and 2138 downregulated DEGs. Further enrichment analysis showed that the upregulated DEGs were enriched in the biological processes mainly involved in signal transduction, transmembrane transport, and innate immune/inflammatory responses. In contrast, the downregulated DEGs were associated with the biological processes mainly including metabolic process, and MHC class I antigen processing and presentation. In addition, 49 of these immune-related DEGs were validated by reverse transcription quantitative PCR (RT-qPCR). Collectively, these data suggest that H9N2 virus infection may enhance the signal transduction, and innate immune responses in chicken DCs, but impair their metabolic functions and antigen-presenting responses, which provide helpful insight into the pathogenesis of H9N2 AIVs in chickens and managing this infection in poultry farms.

This data article reports the global gene expression analysis data of chicken DCs infected with H9N2 avian influenza virus (AIV) compared with mock infection. The differentially expressed genes (DEGs), and the data of GO enrichment analysis and KEGG pathway analysis for DEGs were reported here. In addition, some of these DEGs associated with innate immune response and antigen presentation were also verified by qPCR. The replication of H9N2 AIV in DCs, and the viability kinetic of DCs during H9N2 AIV infection, and the primers for qPCR were also reported in this data article. The data presented here was used on the research article entitled "Transcriptomic profile of chicken bone marrow-derive dendritic cells in response to H9N2 avianinfluenza A virus".

Highlights • The TP1 and TP2 peptides, derived from the TMUV E protein, inhibited TMUV infection. • TP2 exhibited cross-inhibitory activity against JEV. • The TP1 and TP2 peptides blocked TMUV antibody-dependent enhancement (ADE) in duck peripheral blood lymphocytes. • Both peptides interacted with the surface of TMUV, leading to release of viral RNA and interfering with virus:cell binding., The outbreak and spread of Tembusu virus (TMUV) has caused very large losses in the waterfowl-breeding industry since 2010. The viral envelope (E) protein, the principal surface protein of viral particles, plays a vital role in viral entry and fusion. In this study, two peptides derived from domain II (DII) and the stem of the TMUV envelope protein, TP1 and TP2, respectively, were tested for their antiviral activity. TP1 and TP2 inhibited TMUV infection in BHK-21 cells, and their 50% inhibitory concentrations (IC50) were 14.19 mg/L and 7.64 mg/L, respectively. Viral inhibition assays in different cell lines of avian origin showed that the inhibitory effects of TP1 and TP2 are not cell type dependent. Moreover, TP2 also exhibited inhibitory activity against Japanese encephalitis virus (JEV) infection. The two peptides inhibited antibody-mediated TMUV infection of duck peripheral blood lymphocytes. Co-immunoprecipitation assays and indirect enzyme-linked immunosorbent assays (ELISAs) indicated that both peptides interact with the surface of the TMUV virion. RNase digestion assays confirmed the release of viral RNA following incubation with TP1, while incubation with TP1 or TP2 interfered with the binding between TMUV and cells. Taken together, these results show that TP1 and TP2 may be developed into antiviral treatments against TMUV infection.

Duck Tembusu virus (DTMUV) is a newly emerging pathogenic flavivirus that has caused massive economic losses to the duck industry in China. The cellular factors required for DTMUV replication have been poorly studied. The ubiquitin-proteasome system (UPS), the major intracellular proteolytic pathway, mediates diverse cellular processes, including endocytosis and signal transduction, which may be involved in the entry of virus. In the present study, we explored the interplay between DTMUV replication and the UPS in BHK-21 cells and found that treatment with proteasome inhibitor (MG132 and lactacystin) significantly decreased the DTMUV progency at the early infection stage. We further revealed that inhibition of the UPS mainly occurs on the level of viral protein expression and RNA transcription. In addition, using specific siRNAs targeting ubiquitin reduces the production of viral progeny. In the presence of MG132 the staining for the envelope protein of DTMUV was dramatically reduced in comparison with the untreated control cells. Overall, our observations reveal an important role of the UPS in multiple steps of the DTMUV infection cycle and identify the UPS as a potential drug target to modulate the impact of DTMUV infection., Highlights • Treatment with proteasome inhibitor significantly decreased the DTMUV progency. • Inhibition of the UPS mainly occurs on the level of viral protein expression and RNA transcription. • Inhibit the expression of ubiquitin reduces the production of viral progeny.

Background Tembusu virus (TMUV), classified in the genus Flavivirus, causes reduced egg production and neurological problems in poultry. Flavivirus replication depends on the host endoplasmic reticulum (ER) and induces ER stress that leads to activation of the cellular unfolded protein response (UPR), an important signalling pathway that regulates many biological functions involved in viral pathogenesis and innate immunity. However, the mechanism of TMUV-induced UPR activation remains unclear. Results In this study, we systematically investigated the three UPR pathways in TMUV-infected BHK-21 cells. Our results showed that expression of glucose-related protein 78 (GRP78) and GRP94 was upregulated during the course of TMUV infection. We then demonstrated that TMUV activated the PERK pathway in the early stage of infection, resulting in upregulation of ATF4, GADD34 and CHOP, with CHOP induction leading to caspase-3 activation. We also found the IRE1 pathway to be activated, leading to splicing of X box binding protein 1 (XBP1) mRNA and enhanced expression of p58IPK. Finally, we observed increased expression of ATF6 and activity of ER stress-response elements, suggesting stimulation of the ATF6 pathway. In addition, ATF6 pathway activation correlated with the induction of downstream chaperones calnexin, calreticulin, ERp57 and PDI. UPR activity was also observed by the marked elevation in GRP78 and sXBP1 levels in TMUV-infected DF-1 cells. Conclusions This is the first report that TMUV infection-induced ER stress activates three branches of the UPR, and these results lay the foundation for elucidating the pathogenesis of TMUV and understanding the inherent mechanism of TMUV infection as well as the host response.

Newly emerging tembusu virus (TMUV) is a severe threat to poultry industry and causes huge economic losses. Humoral and cell-mediated immunity are both play vital roles in TMUV infection. Up to now, there has been no report on identification of T cell epitopes of the TMUV. In this work, we identified T cell epitopes within TMUV envelope (E) protein using synthesized peptides predicted in silico. A total of ten peptides could stimulate TMUV-specific T cells in murine ELISPOT and duck lymphocyte proliferation assay. Subsequently, DNA vaccine containing these T cell epitopes was constructed (pVAX-T) and the expression of multiepitope protein was confirmed by transfection of BHK-21 cells in vitro. Ducks were administrated intramusclarly to evaluated the immunologic effect of pVAX-T. In ducks immunized with pVAX-T, antibody against TMUV was undetectable, but the expression level of cytokines (IL-2, IL-6, IFN-γ) was upregulated both in peripheral blood lymphocytes and spleen. Furthermore, TMUV challenge revealed that cell-mediated immune response sitmulated by pVAX-T contributed to protection against TMUV infection. The identification of these T cell epitopes will contribute to designing epitope vaccine for preventing infection of TMUV and possibly provide the basis for further studies on cell-mediate immune response activated by TMUV.

Highlights • The newly emerging duck/goose tembusu virus resulted in serious economic loss in China. • GTV E protein DI/II is sufficient to inhibit JS804 infection in a dose-dependent manner. • DI/II binds to cell membrane and this binding blocks goose tembusu virus E protein access to cell membrane., Flavivirus envelope protein locates at the outermost surface of viral particle and mediates virus entry and fusion infection, and domains I and II of E protein play an important role in this process. In this study, we have expressed and purified goose tembusu virus (GTV) E protein domains I and II (DI/II) from E. coli, and tested conceptual approach that purified protein serves as anti-viral reagent. We found that DI/II inhibited GTV JS804 infection in BHK-21 cells in a dose-dependent manner, and this inhibition activity was achieved by binding to cell membrane specifically. Moreover, JS804 treated with DI/II specific anti-serum decreased its infectivity to BHK-21 cells. Taken together, this is first to show that the purified DI/II domain of tembusu virus expressed in E. coli was able to interfere with virus infection, which opens an avenue to develop novel anti-viral regents to prevent and eventually eradicate GTV infection.

Since April 2010, a novel contagious disease in ducks and geese, with egg drop, feed uptake decline and neurological signs, caused by a newly emerged virus has spread around Eastern China. Dissection conducted on the dead geese demonstrated hemorrhage in brain, lung, liver, heart, ovary, and enlarged and necrotic spleen. A new virus, named Goose/Jiangsu/804/2010 (JS804) virus, was isolated in Jiangsu area from geese. Then the virus was re-isolated from the affected geese and replicated well in duck embryo fibroblasts and Vero cells, causing the cytopathic effect. The virus was identified as an enveloped positive stranded RNA virus with a size of approximately 40-60 nm in diameter. The full-length genome of this isolated virus was determined, showing that it is closely related to Tembusu virus (a mosquito-borne Ntaya group flavivirus) than other members of the Flaviviridae based on the data of phylogenetic analyses. Our systematic studies fulfill Koch's postulates precisely, and therefore, the causative agent of geese occurring in Eastern China is a new flavivirus. This is the first report that flavivirus infects not only egg-laying and breeder ducks but also geese. The findings extend our understanding of how the virus spreads and causes disease.

H9N2 Avian influenza virus has repeatedly infected humans and other mammals, which highlights the need to determine the pathogenicity and the corresponding mechanism of this virus for mammals. In this study, we found two H9N2 viruses with similar genetic background but with different pathogenicity in mice. The A/duck/Nanjing/06/2003 (NJ06) virus was highly pathogenic for mice, with a 50% mouse lethal dose of 102.83 50% egg infectious dose, whereas the A/duck/Nanjing/01/1999 (NJ01) virus was low pathogenic for mice, with a 50% mouse lethal dose of >106.81 50% egg infectious dose. Further studies showed that the NJ06 virus grew faster and reached significantly higher titers than NJ01 in vivo and in vitro. Moreover, the NJ06 virus induced more severe lung lesions, and higher levels of inflammatory cellular infiltration and cytokine response in lungs than NJ01 did. However, only twelve different amino acid residues (HA-K157E, NA-A9T, NA-R435K, PB2-T149P, PB2-K627E, PB1-R187K, PA-L548M, PA-M550L, NP-G127E, NP-P277H, NP-D340N, NS1-D171N) were found between the two viruses, and all these residues except for NA-R435K were located in the known functional regions involved in interaction of viral proteins or between the virus and host factors. Summary, our results suggest that multiple amino acid differences may be responsible for the higher pathogenicity of the NJ06 virus for mice, resulting in lethal infection, enhanced viral replication, severe lung lesions, and excessive inflammatory cellular infiltration and cytokine response in lungs. These observations will be helpful for better understanding the pathogenic potential and the corresponding molecular basis of H9N2 viruses that might pose threats to human health in the future.

This study attempts to identify receptor elements for Tembusu virus (TMUV) on DF-1 cells. Using co-immunoprecipitation and virus overlay protein binding assays, we identified a TMUV-binding protein of approximately 70-kDa on DF-1 cell membranes. Mass spectroscopy identified the protein to be heat shock protein (HSP) A9, which was reconfirmed by an anti-HSPA9 antibody. Indirect immunofluorescence demonstrated a significant degree of colocalization between HSPA9 and TMUV on cell surface. Additionally, an antibody against HSPA9 could inhibit TMUV infection in DF-1 cells in a dose-dependent manner. These results might suggest that HSPA9 is a putative receptor for TMUV.

Duck Tembusu virus (DTMUV) is a newly emerging pathogenic flavivirus that has caused massive economic losses to the duck industry in China. DTMUV infection mainly results in significant decreases in egg production in egg-laying ducks within 1-2 weeks post infection. However, information on the comparative protein expression of host tissues in response to DTMUV infection is limited. In the present study, the cellular protein response to DTMUV infection in duck ovarian follicles was analyzed using nano-flow high-performance liquid chromatography-electrospray tandem mass spectrometry. Quantitative proteomic analysis revealed 131 differentially expressed proteins, among which 53 were up regulated and 78 were down regulated. The identified proteins were involved in the regulation of essential processes such as cellular structure and integrity, RNA processing, protein biosynthesis and modification, vesicle transport, signal transduction, and mitochondrial pathway. Some selected proteins that were found to be regulated in DTMUV-infected tissues were screened by quantitative real-time PCR to examine their regulation at the transcriptional level, western blot analysis was used to validate the changes of some selected proteins on translational level. To our knowledge, this study is the first to analyze the proteomic changes in duck ovarian follicles following DTMUV infection. The protein-related information obtained in this study may be useful to understand the host response to DTMUV infection and the inherent mechanism of DTMUV replication and pathogenicity.

A secondary amino-modified mesoporous molecular sieve MCM-41 was obtained by reaction of bis(3-(triethoxysilyl)propyl)amine with MCM-41. The chiral Salen-Mn (III) complex was anchored onto the modified MCM-41 by a multi-step grafting method and two heterogenized catalysts with different Mn contents were obtained. The catalysts were characterized by XRD, N2 adsorption, ICP, FT-IR and DR UV-Vis. Their catalysis on asymmetric epoxidation of several olefins was studied with NaClO and m-CPBA as oxidants respectively. It was found that both the activity and enantioselectivity of the catalysts decreased after the homogeneous catalyst was heterogenized. The reasons resulting in the decrease of catalytic performance were discussed.

A chiral Manganese (III) salen complex was immobilized on the walls of MCM-41 (mobile crystalline material) through the multi-grafting method. The immobilized complex was characterized by XRD, FTIR, UV-Vis, ICP and Nitrogen sorption, and was applied to the asymmetric epoxidation of unfuctionalized alkenes including 1,2-dihydronaphthalene, α-methylstyrene, cis-β-methylstyrene, styrene using NaClO and m-chloroperbenzoic acid (m-CPBA) as oxidants respectively. The immobilized complex showed good activity and enantioselectivity in the epoxidation of 1,2-dihydronaphthalene by using NaClO as oxidant. It could also be run for 4 times in the epoxidation of α-methylstyrene without obvious loss of activity or enantiomeric excess.

Three new salen ligands with carbohydrate moieties were prepared from a salicylaldehyde derivative obtained by reaction of 1,2:5,6-di-O-isopropylidene-α- d -glucofuranose with 3-tert-butyl-5-(chloro-methyl)-2-hydroxybenzaldehyde. These ligands were coordinated with Mn(III) to give three chiral salen–Mn(III) complexes. The complexes were characterized and employed in the asymmetric epoxidation of unfunctionalized alkenes. Catalytic results showed that although there are no chiral groups on the diimine bridge, these complexes had some enantioselectivity, which indicates the carbohydrate moiety has an asymmetric inducing effect in the epoxidation reaction.

UNLABELLED Flavivirus envelope protein locates at the outermost surface of viral particle and mediates virus entry and fusion infection, and plays an important role in eliciting neutralizing antibody. In this study, goose tembusu virus (GTV) E protein was expressed and purified from Escherichia coli. The immunological changes and protection efficiency of this protein serving as recombinant subunit vaccine were then tested. In ducks, recombinant E protein markedly elicited specific neutralizing antibody, stimulated the secretion of IL-2, TNF-α and IFN-γ cytokines and promoted the proliferation of lymphocytes. Additionally, population of soluble CD4 and soluble CD8 molecules in sera significantly increased in response to this antigen. Moreover, virus challenge revealed that recombinant E protein contributed to protection against GTV challenge. Taken together, the data indicate that recombinant GTV E protein could induce significant humoral and cell-mediated responses and that recombinant E protein can serve as a potential subunit vaccine candidate against GTV. SIGNIFICANCE AND IMPACT OF THE STUDY The sudden outbreak and quick spread of newly emerging goose tembusu virus (GTV) have resulted in serious economic loss. There is no effective commercial vaccine or reasonably available control measure so far. In this study, GTV E protein was expressed and purified from Escherichia coli, and it was found that recombinant E protein could induce significant humoral and cell-mediated responses. It indicated that recombinant E protein can serve as a potential subunit vaccine candidate against GTV.

An unsymmetrical manganese (III) salen complex was anchored onto the surface of MCM-41 by a multi-step grafting method. The resulting material was characterized by FT-IR, X-ray diffraction, DR UV–Vis, nitrogen adsorption and ICP. The catalytic activity of the immobilized catalyst was tested in the aerobic epoxidation of olefins. The immobilized catalyst showed relatively high activity and epoxide selectivity in the aerobic epoxidation of cyclohexene, styrene, 1,2-dihydronaphthalene and α-methylstyrene. The immobilized catalyst can be reused six times. However, the conversion of cyclohexene decreased from more than 99% in the first run to 73.7% in the sixth, while 64.8% of manganese leaching was observed.

Heat shock protein A9 (HSPA9), a member of the heat shock protein family, is a putative receptor for Tembusu virus (TMUV). By using Western blot and co-immunoprecipitation assays, E protein domains I and II were identified as the functional domains that facilitate HSPA9 binding. Twenty-five overlapping peptides covering domain I and domain II sequences were synthesized and analyzed by using an HSPA9 binding assay. Two peptides showed the capability of binding to HSPA9. Dot blot assay of truncated peptides indicated that amino acid residues 19 to 22 and 245 to 252 of E protein constitute the minimal motifs required for TMUV binding to HSPA9. Importantly, peptides harboring those two minimal motifs could effectively inhibit TMUV infection. Our results provide insight into TMUV-receptor interaction, thereby creating opportunities for elucidating the mechanism of TMUV entry.

Avian tembusu virus (TMUV), which was first identified in eastern China, is an emerging virus causing serious economic losses in the Chinese poultry industry. Here, we report the complete genome sequence of goose tembusu virus strain JS804, isolated from Jiangnan white geese with severe neurological signs. The genome of JS804 is 10,990 nucleotides (nt) in length and contains a single open reading frame encoding a putative polyprotein of 3,425 amino acids. Research of the whole sequence of tembusu virus will help us to understand further the molecular and evolutionary characteristics and pathogenesis of this virus.

With the consideration of the system features, an indicator system of information assurance for aeronautical information system is established. The indicator system is based on SSE (Systems Security Engineering) theory, and DSC (Dynamic System Control) method. The indicator system is established from four aspects, stratagem, technique, management and engineering. Fuzzy-AHP (Analytic Hierarchy Process) is used to evaluate the static indicators. A quite good result is got.