Porcine FcγRIIb mediates enhancement of porcine reproductive and respiratory syndrome virus (PRRSV) infection

Antibody-dependent enhancement (ADE) of virus infection caused by the uptake of virus-antibody complexes by FcγRs is a significant obstacle to the development of effective vaccines to control certain human and animal viral diseases. The activation FcγRs, including FcγRI and FcγRIIa have been shown to mediate ADE infection of virus. In the present paper, we showed that pocine FcγRIIb, an inhibitory FcγR, mediates ADE of PRRSV infection. Stable Marc-145 cell lines expressing poFcγRIIb (Marc-poFcγRII) were established. The relative yield of progeny virus was significantly increased in the presence of sub-neutralization anti-PRRSV antibody. The Fab fragment and normal porcine sera had no effect. Anti-poFcγRII antibody inhibited the enhancement of infection when cells were infected in the presence of anti-PRRSV antibody, but not when cells were infected in the absence of antibody. These results indicate that enhancement of infection in these cells by anti-PRRSV virus antibody is FcγRII-mediated. Identification of the inhibitory FcγR mediating ADE infection should expand our understanding of the mechanisms of pathogenesis for a broad range of infectious diseases and may open many approaches for improvements to the treatment and prevention of such diseases.     

Inhibition of hepatitis B virus replication in cultured cells and in vivo using 2′-O-guanidinopropyl modified siRNAs

Silencing hepatitis B virus (HBV) gene expression with exogenous activators of the RNA interference (RNAi) pathway has shown promise as a new mode of treating infection with the virus. However, optimizing efficacy, specificity, pharmacokinetics and stability of RNAi activators remains a priority before clinical application of this promising therapeutic approach is realised. Chemical modification of synthetic short interfering RNAs (siRNAs) provides the means to address these goals. This study aimed to assess the benefits of incorporating nucleotides with 2′-O-guanidinopropyl (GP) modifications into siRNAs that target HBV. Single GP residues were incorporated at nucleotide positions from 2 to 21 of the antisense strand of a previously characterised effective antiHBV siRNA. When tested in cultured cells, siRNAs with GP moieties at selected positions improved silencing efficacy. Stability of chemically modified siRNAs in 80% serum was moderately improved and better silencing effects were observed without evidence for toxicity or induction of an interferon response. Moreover, partially complementary target sequences were less susceptible to silencing by siRNAs with GP residues located in the seed region. Hydrodynamic co-injection of siRNAs with a replication-competent HBV plasmid resulted in highly effective knock down of markers of viral replication in mice. Evidence for improved efficacy, reduced off target effects and good silencing in vivo indicate that GP-modifications of siRNAs may be used to enhance their therapeutic utility.     

Porcine reproductive and respiratory syndrome virus induces interleukin-15 through the NF-κB signaling pathway

Porcine reproductive and respiratory syndrome virus (PRRSV) mainly infects macrophages/dendritic cells and modulates cytokine expression in these cells. Interleukin-15 (IL-15) is a pleiotropic cytokine involved in wide range of biological activities. It has been shown to be essential for the generation, activation, and proliferation of NK and NKT cells and for the survival and activation of CD8(+) effector and memory T cells. In this study, we discovered that PRRSV infection upregulated IL-15 production at both the mRNA and protein levels in porcine alveolar macrophages (PAMs), blood monocyte-derived macrophages (BMo), and monocyte-derived dendritic cells (DCs). We subsequently demonstrated that the NF-κB signaling pathway was essential for PRRSV infection-induced IL-15 production. First, addition of an NF-κB inhibitor drastically reduced PRRSV infection-induced IL-15 production. We then found that NF-κB was indeed activated upon PRRSV infection, as evidenced by IκB phosphorylation and degradation. Moreover, we revealed an NF-κB binding motif in the cloned porcine IL-15 (pIL-15) promoter, deletion of which abrogated the pIL-15 promoter activity in PRRSV-infected alveolar macrophages. In addition, we demonstrated that PRRSV nucleocapsid (N) protein had the ability to induce IL-15 production in porcine alveolar macrophage cell line CRL2843 by transient transfection, which was mediated by its multiple motifs, and it also activated NF-κB. These data indicated that PRRSV infection-induced IL-15 production was likely through PRRSV N protein-mediated NF-κB activation. Our findings provide new insights into the molecular mechanisms underling the IL-15 production induced by PRRSV infection.     

Inhibition of Rac and Rac-linked functions by 8-oxo-2′-deoxyguanosine in murine macrophages

Rac is a protein involved in the various functions of macrophages (Mφ), including the production of reactive oxygen species (ROS), phagocytosis, chemotaxis and the secretion of cytokines (such as γ-INF). This study tested the effects of nucleosides containing 8-oxoguanine(8-hydroxyguanine) such as 8-oxo-2′-guanosine (8-oxoG) or 8-oxo-2′-deoxyguanosine (8-oxodG), on Rac and the above-listed Rac-associated functions of Mφ using mouse peritoneal Mφ (MpMφ). It is reported that 8-oxodG was able to effectively inhibit Rac and the Rac-associated functions of MpMφ. Compared to 8-oxodG, 8-oxoG showed negligible effects. Furthermore, normal nucleosides such as deoxyguanosine (dG), guanosine (G) and adenosine (A) did not exert any effects. These results suggest that 8-oxodG could be used as a potential tool to modulate the functions of Mφ that are intimately related to various pathological processes.     

The nonstructural protein 1 papain-like cysteine protease was necessary for porcine reproductive and respiratory syndrome virus nonstructural protein 1 to inhibit interferon-β induction

Porcine reproductive and respiratory syndrome virus nonstructural protein 1 (nsp1) could be auto-cleaved into nsp1α and nsp1β, both of which had the papain-like cysteine protease activities. Previous studies have shown that porcine reproductive and respiratory syndrome virus nsp1 was an interferon (IFN) antagonist. However, the mechanism by which nsp1 inhibited IFN-β production was unclear. Here, we used site-directed mutagenesis that inactivated the papain-like cysteine protease activities of nsp1 to explore whether the papain-like cysteine protease activities were required for nsp1 to disrupt IFN-β production. The results showed that mutations that inactivated papain-like cysteine protease activity of nsp1α made nsp1 lose its IFN antagonism activity, whereas mutations that inactivated papain-like cysteine protease activity of nsp1β did not influence the IFN antagonism activity of nsp1. In conclusion, our present work indicated that the papain-like cysteine protease activity of nsp1α was necessary for nsp1 to inhibit IFN-β induction.     

Genome organization and variation in the 3′-partial sequence of garlic latent virus in China

Ten different isolates of a carlavirus were detected by degenerate PCR from 12 garlic samples collected from 6 provinces in China, and the complete genome sequence of the Zhejiang isolate ZJ1 and 3′-terminal sequences of 9 other isolates were determined. The RNA genome of isolate ZJ1 consisted of 8363nts excluding the 3′-poly (A) tail, and the genome organization was similar to other carlaviruses with 6 open reading frames encoding a replicase, TGB1, TGB2, TGB3, CP and NABP respectively. Sequence comparisons showed that all 10 isolates were Garlic latent virus (GarLV). The variations in the TGB2, TGB3 and NABP were more significant than those in the CP. High homology was also detected between those isolates and Shallot latent virus (ShLV). Phylogenetic analysis suggested that GarLV isolates from garlic can be divided into 4 main groups and Chinese isolates belonged to each group. This is the first reported molecular analysis of members of the genus Carlavirus in China.     

Diversity and evolution of samara in angiosperm北大核心CSCD

Samara (winged fruit) can be dispersed easily by wind and may be a crucial factor for angiosperm spread and diversification. In a narrow sense, a samara is an indehiscent dry fruit with wing(s) developed from fruit pericarp, while in a broad sense samaras also include all winged fruits with wings developed from both pericarp and peri-anth or bracts. According to the wing shape and growth patterns of samaras, we divided samaras into six types, i.e. single-winged, lanceolate-winged, rib-winged, sepal-winged, bract-winged, and perigynous samaras. Perigynous samaras can be further classified into two forms, i.e. round-winged and butterfly-winged samaras. Accordingly, the aerodynamic behavior of samaras can be classified into five types, autogyro, rolling autogyro, undulator, helicopter, and tumbler. The rib-winged and round-winged samaras can be found in Laurales, a basal angiosperm, and may represent the primitive type of early samaras. In the derived clades, samaras evolved enlarged but unequal wings and decreased wing loading (the ratio of fruit weight to wing size), which is likely an adaptation to gentle wind and secondary dispersal through water or ground wind. The wings of some samaras (such as sepal-winged and bract-winged samaras) may have multiple functions including wind dispersal, physical defense for the seeds, and adjust seed germination strategy. The pantropical family Malpighiaceae is extraordinarily rich in samara types, which is likely related to its multiple inter-continent dispersal in history, which is known as “Malpighiaceae Route”. Therefore, Malpighiaceae can be used as a model system for the studies on samara adaptation and evolution. We identified the following issues that deserve further examination in future studies using both ecological and evo-devo methods: 1) the adaption of different types of samaras in dispersal processes, 2) the molecular and developmental mechanism of sepal- and bract-wings, and 3) the evolution of samara types and their effects on angiosperm diversification. © 2018 Editorial Office of Chinese Journal of Plant Ecology. All rights reserved.     

Anatomical characteristics of xylem in tree rings and its relationship with environments北大核心CSCD

The anatomical traits of xylem are the characteristics of tree rings at the cellular and subcellular scales, and are often reflection of environmental signals. Studying the relationships between anatomical traits of xylem and environmental change not only provide physiological explanations to the statistics in dendroclimatology, but can also provide a new vision for studying the adaptation process and response strategies of tree growth to climate change. In this paper, with the relationships between the anatomical characteristics of xylem in tree-rings (cell chronology) and climate change as a main thread, we first outline the basic principles and mechanisms of wood anatomical features to record environmental signals, and expounded the basic methods involved in the process of xylem anatomy. Secondly, we discuss the relationship between the anatomical features of xylem and climate factors. We then propose the following as possible directions of future research based on the existing knowledge gap in the topical area: (1) to explore the temporal and spatial variations in the anatomical characteristics of xylem in tree-rings along radial and tangential directions and the relationships with environmental changes; (2) to explore the threshold of tree growth response to environmental plasticity and adaptation processes; (3) to assess the synergistic and antagonistic effects as well as the formation mechanisms of climate response among different tree-ring proxies, and to determine the specific roles and contributions of major climatic factors during different periods of tree-ring formation.     

Constraint line methods and the applications in ecology北大核心CSCD

With increasing data availability in the big data era, many traditional statistical analyses based on the mean or median are insufficient or inappropriate to elucidate the complex patterns of variation. This is particularly the case when multiple factors are involved and the bivariate scatter occurs as scatter clouds. In such circumstances, constraint line (or envelope) method could be an alternative and effective tool to extract the data boundaries, thus improves our understanding of the complex relationships between limiting factor and response factor. Here, we synthesize the major findings and achievements in the field of applying the constraint line method in ecology. Specifically, we first describe the history and development of the constraint line method. We then discuss the techniques to establish the constraint lines with examples, and discuss the applications and implications of the constraint lines in species distribution, population performance, and optimization problem. We suggest simultaneously application of both constraint lines and regression techniques to the same datasets to achieve a comprehensive understanding of ecological process and underlying mechanisms. Such combined methods should be used with special attention to the role of spatial heterogeneity and scale dependency. We also discuss in detail the potential applicability of the constraint line method in studying the linkages between ecosystem services, and land system design.     

Effects of experimental warming on plant reproductive phenology in Xizang alpine meadow北大核心CSCD

Aims: Climate warming strongly influences reproductive phenology of plants in alpine and arctic ecosystems. Here we focus on phenological shifts caused by warming in a typical alpine meadow on the Qinghai-Xizang Plateau. Our objective was to explore phenological responses of alpine plant species to experimental warming. Methods: Passive warming was achieved using open-top chambers (OTCs). The treatments included control (C), and four levels of warming (T1, T2, T3, T4). We selected Kobresia pygmaea, Potentilla saundersiana, Potentilla cuneata, Stipa purpurea, Festuca coelestis and Youngia simulatrix as the focal species. Plant phenology was scored every 3-5 days in the growing season. The reproductive phenology phases of each species were estimated through fitting the phenological scores to the Richards function. Important findings: Under soil water stress caused by warming, most plants in the alpine meadow advanced or delayed their reproductive events. As a result, warming significantly delayed phenological development of K. pygmaea. Warming significantly advanced reproductive phenology of P. saundersiana, S. purpurea and F. coelestis, but not of P. cuneata and Y. simulatrix. In addition, warming significantly shortened the average flowering duration of alpine plant species. The potentially warmer and drier growing seasons under climate change may shift the reproductive phenology of the alpine systems in similar pattern.     

An update of the vegetation classification in China北大核心CSCD

The vegetation classification in China was updated by referring to recent advances for vegetation classification worldwide and on the basis of our former paper Recognition and Proposal on the Vegetation Classification System of China (hereafter, \"Recognition and Proposal\"). In this revision, the criteria for vegetation classification were discussed and unified, and a quantified standard for high, medium, and low level units in a new hierarchical classification scheme was supplemented. Compared with the old classification scheme in \"Recognition and Proposal\", the units at the level of vegetation type were substantially changed. Finally, in order to improve mutual communication among international peers, a comparison was carried out between the new revised scheme and each of International Classification and Mapping of Vegetation of UNESCO, The National Vegetation Classification Standard of United States, and The Braun-Blanquet Classification Scheme.     

Inhibition of HCV translation by disrupting the structure and interactions of the viral CRE and 3′ X-tail

A phylogenetically conserved RNA structure within the NS5B coding region of hepatitis C virus functions as a cis-replicating element (CRE). Integrity of this CRE, designated SL9266 (alternatively 5BSL3.2), is critical for genome replication. SL9266 forms the core of an extended pseudoknot, designated SL9266/PK, involving long distance RNA–RNA interactions between unpaired loops of SL9266 and distal regions of the genome. Previous studies demonstrated that SL9266/PK is dynamic, with ‘open’ and ‘closed’ conformations predicted to have distinct functions during virus replication. Using a combination of site-directed mutagenesis and locked nucleic acids (LNA) complementary to defined domains of SL9266 and its interacting regions, we have explored the influence of this structure on genome translation and replication. We demonstrate that LNAs which block formation of the closed conformation inhibit genome translation. Inhibition was at least partly independent of the initiation mechanism, whether driven by homologous or heterologous internal ribosome entry sites or from a capped message. Provision of SL9266/PK in trans relieved translational inhibition, and mutational analysis implied a mechanism in which the closed conformation recruits a cellular factor that would otherwise suppresses translation. We propose that SL9266/PK functions as a temporal switch, modulating the mutually incompatible processes of translation and replication.     

Inhibition of the synthesis of polyamines and macromolecules by 5′-methylthioadenosine and 5′-alkylthiotubercidins in BHK21 cells

5′-Methylthioadenosine and four 5′-alkylthiotubercidins were tested for their ability to inhibit polyamine synthesis in vitro and to decrease polyamine concentration and prevent growth of baby-hamster-kidney (BHK21) cells. 5′-Methylthioadenosine and 5′-methylthiotubercidin decreased the activity of spermidine synthase from brain to roughly the same extent, whereas brain spermine synthase was much more strongly inhibited by 5′-methylthioadenosine compared with 5′-methylthiotubercidin. These nucleoside derivatives also inhibited the growth of BHK21 cells and increased the concentration of putrescine. 5′-Methylthioadenosine decreased cellular spermine concentration, whereas 5′-methylthiotubercidin lowered the concentration of spermidine. The activities of ornithine decarboxylase and S-adenosylmethionine decarboxylase were enhanced in cells grown in the presence of 5′-methylthiotubercidin. The growth inhibition produced by these nucleoside derivatives was not reversed by exogenous spermidine or spermine. 5′-Ethylthiotubercidin, 5′-propylthiotubercidin and 5′-isopropylthiotubercidin did not appreciably inhibit spermidine or spermine synthase in vitro or decrease the cellular polyamine content, but effectively prevented the growth of BHK21 cells. All nucleoside derivatives at concentrations of 0.2–1 mm caused a rapid inhibition of protein synthesis. It is concluded that the growth inhibition produced by 5′-methylthioadenosine and 5′-alkylthiotubercidins was not primarily due to polyamine depletion but other target sites, for instance the cellular nucleotide pool, cell membranes etc. must be considered.     

Application and comparison of remote sensing GPP models with multi-site data in China北大核心CSCD

Aims Estimation of gross primary productivity (GPP) of vegetation at the global and regional scales is important for understanding the carbon cycle of terrestrial ecosystems. Due to the heterogeneous nature of land surface, measurements at the site level cannot be directly up-scaled to the regional scale. Remote sensing has been widely used as a tool for up-saling GPP by integrating the land surface observations with spatial vegetation patterns. Although there have been many models based on light use efficiency and remote sensing data for simulating terrestrial ecosystem GPP, those models depend much on meteorological data; use of different sources of meteorological datasets often results in divergent outputs, leading to uncertainties in the simulation results. In this study, we examines the feasibility of using two GPP models driven by remote sensing data for estimating regional GPP across different vegetation types. Methods Two GPP models were tested in this study, including the Temperature and Greenness Model (TG) and the Vegetation Index Model (VI), based on remote sensing data and flux data from the China flux network (ChinaFLUX) for different vegatation types for the period 2003-2005. The study sites consist of eight ecological stations located in Xilingol (grassland), Changbaishan (mixed broadleaf-conifer forest), Haibei (shrubland), Yucheng (cropland), Damxung (alpine meadow), Qianyanzhou (evergreen needle-leaved forest), Dinghushan (evergreen broad-leaved forest), and Xishuangbanna (evergreen broad-leaved forest), respectively. Important findings All the remote sensing parameters employed by the TG and VI models had good relationships with the observed GPP, with the values of coefficient of determination, R2, exceeding 0.67 for majority of the study sites. However, the root mean square errors (RMSEs) varied greatly among the study sites: the RMSE of TG ranged from 0.29 to 6.40 g·m-2·d-1, and that of VI ranged from 0.31 to 7.09 g·m-2·d-1, respectively. The photosynthetic conversion coefficients m and a can be up-scaled to a regional scale based on their relationships with the annual average nighttime land surface temperature (LST), with 79% variations in m and 58% of variations in a being explainable in the up-scaling. The correlations between the simulated outputs of both TG and VI and the measured values were mostly high, with the values of correlation coefficient, r, ranging from 0.06 in the TG model and 0.13 in the VI model at the Xishuangbanna site, to 0.94 in the TG model and 0.89 in the VI model at the Haibei site. In general, the TG model performed better than the VI model, especially at sites with high elevation and that are mainly limited by temperature. Both models had potential to be applied at a regional scale in China.     

Association of HLA classⅠand classⅡgenes with severe acute respiratory syndrome in the northern Chinese population

Severe acute respiratory syndrome (SARS) was a major epidemic at the beginning of the 21^st century. This highly infectious disease is caused by a novel coronavirus (SARS-CoV), whose immune reaction is still not completely understood. This study described the genetic patterns of HLA-A, -B, and -DRB1 loci in patients from Beijing who survived SARS, and examined whether an association between HLA genes and susceptibility/resistance to SARS exists. A total of 148 Chinese Han SARS survivors were recruited to donate convalescent plasma in 2003. HLA low-resolution genotyping was carried out using PCR-SSP. Allele frequencies were compared with published frequencies of HLA alleles from 11 755 unrelated northern Chinese Han bone marrow donors by Fisher''s exact test. In this cohort, 13, 25 and 13 alleles were observed at HLA-A, -B, and -DRB1 loci respectively. Fisher''s exact tests revealed four alleles (A*26, DRB1*04, DRB1*09, and DRB1*16) that showed a nominal association significance with the SARS virus (P<0.05), yet none of these associations remained significant after correction. Our study suggests that HLA polymorphisms were unlikely to have contributed significantly to either the susceptibility or resistance to the SARS-Cov infection in patients who survived SARS in the Northern Chinese population, thus leaving an open question for future studies into a possible association HLA class Ⅰ and class Ⅱ genes with SARS in patients who were unable to survive the infection.     

Association of HLA classⅠand classⅡgenes with severe acute respiratory syndrome in the northern Chinese population

Severe acute respiratory syndrome (SARS) was a major epidemic at the beginning of the 21^st century. This highly infectious disease is caused by a novel coronavirus (SARS-CoV), whose immune reaction is still not completely understood. This study described the genetic patterns of HLA-A, -B, and -DRB1 loci in patients from Beijing who survived SARS, and examined whether an association between HLA genes and susceptibility/resistance to SARS exists. A total of 148 Chinese Han SARS survivors were recruited to donate convalescent plasma in 2003. HLA low-resolution genotyping was carried out using PCR-SSP. Allele frequencies were compared with published frequencies of HLA alleles from 11 755 unrelated northern Chinese Han bone marrow donors by Fisher's exact test. In this cohort, 13, 25 and 13 alleles were observed at HLA-A, -B, and -DRB1 loci respectively. Fisher's exact tests revealed four alleles (A*26, DRB1*04, DRB1*09, and DRB1*16) that showed a nominal association significance with the SARS virus (P<0.05), yet none of these associations remained significant after correction. Our study suggests that HLA polymorphisms were unlikely to have contributed significantly to either the susceptibility or resistance to the SARS-Cov infection in patients who survived SARS in the Northern Chinese population, thus leaving an open question for future studies into a possible association HLA class Ⅰ and class Ⅱ genes with SARS in patients who were unable to survive the infection.