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Trichiurus japonicus is an economically important fish that ranks 11th in global marine fish capture production. However, the reproductive characteristics of this fish have undergone notable changes in recent decades, potentially affecting the quality of offspring and sustainable utilization. To improve our understanding of the physiological regulation of maturation in T. japonicus, untargeted liquid chromatography mass spectrometry was utilized to identify the small molecules that characterize the comprehensive metabolic profiles of ovaries during ovary development from stage II to stage IV. According to the results of OPLS-DA, the ovarian metabolite profiles of the three developmental stages were separated. The concentrations of 124 and 100 metabolites were significantly altered between stage II vs. III and III vs. IV, respectively. Lipids and lipid-like molecules accounted for the largest proportion of the altered metabolites, followed by amino acids, peptides, and analogues. The significantly altered metabolites-enriched pathways differed slightly between stages II and III and stages III and IV. Steroid-related pathways were heavily affected during stages II to III, while significantly altered metabolites from stages III to IV were involved in oocyte-maturation-related pathways. Through metabonomics analysis, potentially important metabolic pathways and metabolites between different ovarian stages were detected, providing basic information for further investigation of maturation mechanisms in wild fish. [ABSTRACT FROM AUTHOR]

Many cells in vivo have their inherent motions, which involve numerous biochemical and biophysical signals synergistically regulating cell behavior and function. However, existing methods offer little information about the concurrently chemical and physical responses of dynamically pulsing cells. Here, we report a soft electrode with an electrospun poly(3,4‐ethylenedioxythiophene) (PEDOT)‐based nanomesh to fully comply with spontaneous motions of cells. Moreover, this electrode demonstrated excellent electrical conductivity, electrochemical performance and cellular biocompatibility. Cardiomyocytes cultured thereon exhibited autonomous and rhythmic contractility, and synchronously induced mechanical deformation of the underlying electrode, which allowed real‐time monitoring of nitric oxide release and electrophysiological activity of cardiomyocytes. This work provides a promising way toward recording chemical and electrical signals of biological systems with their natural motions. [ABSTRACT FROM AUTHOR]

Many cells in vivo have their inherent motions, which involve numerous biochemical and biophysical signals synergistically regulating cell behavior and function. However, existing methods offer little information about the concurrently chemical and physical responses of dynamically pulsing cells. Here, we report a soft electrode with an electrospun poly(3,4‐ethylenedioxythiophene) (PEDOT)‐based nanomesh to fully comply with spontaneous motions of cells. Moreover, this electrode demonstrated excellent electrical conductivity, electrochemical performance and cellular biocompatibility. Cardiomyocytes cultured thereon exhibited autonomous and rhythmic contractility, and synchronously induced mechanical deformation of the underlying electrode, which allowed real‐time monitoring of nitric oxide release and electrophysiological activity of cardiomyocytes. This work provides a promising way toward recording chemical and electrical signals of biological systems with their natural motions. [ABSTRACT FROM AUTHOR]

Using the mixed‐ligand method, a novel coordination polymer containing Co(II) of [Co(ditb)(tdc)]n·5nH2O (1) was prepared by reacting rigid 2,5‐thiophenedicarboxylic acid with flexible 1,4‐di(1H‐1,2,4‐triazol‐1‐yl)butane using the slow evaporation method. Active samples of 1 (1a for short) showed good stability and the ability to capture CO2. It can be used as a potential heterogeneous catalyst for chemical immobilization of epoxide and CO2 to form cyclic carbonates. In addition, catalyst 1a retained its original structure after four times of reuse. Its biological properties were assessed against Mycobacterium tuberculosis (M.TB) both in vivo and in vitro. First, the bactericidal ability of the compound against M.TB was measured by a sterilization curve. Then, the expression of pckA gene related to M.TB survival was detected using reverse transcription‐polymerase chain reaction. For the in vivo study, the colony‐forming unit determination was performed for evaluating the anti‐bacterial effect of the compound, and the enzyme‐linked immunosorbent assay was used for the inflammatory cytokine measurement in vivo. Through molecular docking simulations, all three types of polar atoms on the complex were confirmed to have chances of interacting with the protein. [ABSTRACT FROM AUTHOR]

Bot flies (Oestridae) are obligate endoparasites of mammals, and their extraordinary diversification is of great importance in understanding the evolution of parasitism. However, evolutionary analysis of Oestridae has long been impeded by lack of information. Here, the first three mitochondrial genomes of nasal bot flies (Cephalopina titillator , Cephenemyia trompe and Rhinoestrus usbekistanicus) and a comparative mitochondrial genomic analysis between subfamilies of Oestridae are presented. Contrasting to many other parasites, mitochondrial genomes of oestrids are conserved in structure, and genes have retained the same order and direction as the ancestral insect mitochondrial genome. Nucleotide composition is highly heterogenous, with Gasterophilinae possessing highest GC content and smallest genomic size. Mitochondrial evolutionary rates vary considerably, with Hypodermatinae and Oestrinae exhibiting a faster average rate than Cuterebrinae and Gasterophilinae. In addition, the first phylogenomic analysis covering all four bot fly subfamilies was conducted, supporting monophyly of Oestridae and a sister-group relationship of Hypodermatinae and Oestrinae. The only topological ambiguity is Cuterebrinae being a sister-group of either (Hypodermatinae + Oestridae) or Gasterophilinae. Thus, we suggest that mitochondrial genomes carry a great potential for phylogenetic analysis of Oestridae, and more information of Cuterebrinae is needed to illuminate the early evolutionary radiation and parasite-host coevolution of bot flies. [ABSTRACT FROM AUTHOR]

Abstract By introduction of trifluoromethyl and phenyl groups to 2-(2-thienyl)pyridine (thp), four new phosphorescent bis-cyclometalated iridium(III) complexes, (thp) 2 Ir(tpip), (cf 3 thp) 2 Ir(tpip), (cf 3 btp) 2 Ir(tpip) and (3-cf 3 btp) 2 Ir(tpip) (cf 3 thp = 2-(thiophen-2-yl)-4-(trifluoromethyl)pyridine, cf 3 btp = 2-(benzo[ b ]thiophen-2-yl)-4- (trifluoromethyl)pyridine, 3-cf 3 btp = 2-(benzo[ b ]thiophen-3-yl)-4-(trifluoromethyl)pyridine, tpip = tetraphenylimidodiphosphinate), were synthesized and fully characterized. The density functional theory and time-dependent DFT calculations show that the frontier orbitals are mainly localized in the Ir(III) ion and the cyclometalated ligands. Thus, the photophysical properties were dominated by the cyclometalated C∧N ligand, attributed to 3MLCT and 3ILCT transition. These Ir(III) complexes emit in the yellow-to-deep red region with photoluminescence quantum yields in the range 40.5–86.4% in CH 2 Cl 2 solutions at 298 K. The organic light-emitting diodes (OLEDs) using (thp) 2 Ir(tpip), (cf 3 thp) 2 Ir(tpip) and (cf 3 btp) 2 Ir(tpip) as yellow, orange and deep red emitters display good electroluminescent performance with low efficiency roll-off. Notably, the device based on (cf 3 thp) 2 Ir(tpip) possesses very high EL efficiencies with the maximum luminance efficiency and external quantum efficiency (EQE) of 53.9 cd A−1 and 17.9%, respectively. Furthermore, the EQE for this complex could be still retained as 15.4% at a luminance of 1000 cd m−2. Graphical abstract Image 1 Highlights • Four new phosphorescent iridium(III) complexes based on thienylpyridine derivatives were synthesized. • These iridium(III) complexes emit in the yellow-to-deep red region with photoluminescence quantum yields of 40.5–86.4%. • The yellow-to-deep red OLEDs have good performance with low efficiency roll-off. • The best device showed an EQE max of 17.9%, and the EQE remained as 15.4% at a luminance of 1000 cd m−2. [ABSTRACT FROM AUTHOR]

Abstract: Hypercholesterolemia is a dominant risk factor for the development and progression of atherosclerosis and cardiovascular diseases. Natural compounds have been proved to be useful in lowering serum cholesterol to slow down the progression of cardiovascular diseases. Pueraria lobata is employed clinically to treat cardiovascular diseases in China. In the present study, the atheroscleroprotective potential of the herb''s major active compound, puerarin, was investigated by monitoring serum lipid profile and major enzyme expressions on cholesterol homeostasis in Sprague–Dawley rats fed with control diet, hypercholesterolmic diet or hypercholesterolmic diet plus administration of puerarin (300 mg/kg/day, p.o.) for 4 weeks. Puerarin markedly attenuated the increased total cholesterol induced by hypercholesterolmic diet in both serum and liver. It caused a significant reduction in the atherogenic index. Expression of mRNA for hepatic 7α-hydroxylase (CYP7A1) was significantly enhanced but not for those of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) and lanosterol 14α-demethylase (CYP51). To further explore the atheroscleroprotective potential of puerarin, acetylcholine induced endothelium-dependent vasorelaxation and endothelial nitric oxide synthase (eNOS) expression on isolated thoracic aortas were analyzed. Animals administered with puerarin suppressed the hypercholesterolemic diet induced impairment of eNOS expression, whereas there was no significant difference in the endothelium-dependent vasorelaxation among various groups of animals. These data indicated that puerarin reduced the atherogenic properties of dietary cholesterol in rats. Its hypocholesterolemic function may be due to the promotion of cholesterol and bile acids excretion in liver. Whether puerarin targets directly on cholesterol homeostasis or both cholesterol homeostasis and endothelial function remains to be determined. [Copyright &y& Elsevier]

Mitogenomes have been widely used for phylogenetic reconstruction of various Dipteran groups, but specifically for chironomid, they have not been carried out to resolve the relationships. Diamesinae (Diptera: Chironomidae) are important bioindicators for freshwater ecosystem monitoring, but its evolutionary history remains uncertain for lack of information. Here, coupled with one previously published and 30 new mitogenomes of Diamesinae, we carried out comparative mitogenomic analysis and phylogenetic analysis. Mitogenomes of Diamesinae were conserved in structure, and all genes arranged in the same order as the ancestral insect mitogenome. All protein‐coding genes in Diamesinae were under stronger purifying selection than those of other nonbiting midge species, which may exhibit signs of adaptation to life at cold living conditions. Phylogenetic analyses strongly supported the monophyly of Diamesinae, with Boreheptagyiini deeply nested within Diamesini. In addition, phylogenetic relationship of selected six genera was resolved, except Sympotthastia remained unstable. Our study revealed that the mitogenomes of Diamesinae are highly conserved, and they are practically useful for phylogenetic inference. [ABSTRACT FROM AUTHOR]

Evolutionary analysis of Prodiamesinae has long been impeded by lack of information, and its phylogenetic relationship with Orthocladiinae remains questionable. Here, ten complete mitochondrial genomes (mitogenomes) of Orthocladiinae sensu lato were newly sequenced, including three Prodiamesinae species and seven Orthocladiinae species. Coupled with published mitogenomes, a total of 12 mitogenomes of Orthocladiinae sensu lato were selected for a comparative mitogenomic analysis and phylogenetic reconstruction. Mitogenomes of Orthocladiinae sensu lato are conserved in structure, and all genes arrange the same gene order as the ancestral insect mitogenome. Nucleotide composition is highly biased, and the control region displayed the highest A + T content. All protein‐coding genes are under purifying selection, and the ATP8 evolves at the fastest rate. In addition, the mitogenomes of Orthocladiinae sensu lato are highly conserved, and they are practically useful for phylogenetic inference, suggesting a re‐classification of Orthocladiinae by sinking Prodiamesinae as a subgroup of Orthocladiinae. [ABSTRACT FROM AUTHOR]

Background: Asthma is a common complex chronic, inflammatory polygenic disease with heterogeneous manifestations, affecting individuals of all age groups and posing an immense burden on healthcare resources. A number of studies have identified the association between a disintegrin and metalloprotease 33 (ADAM33) polymorphisms and asthma risk, however, the results still remain inconclusive. The objective of the present study was to identify the effect of ADAM33 variants in asthma susceptibility.Methods: Eligible case-control studies published between January 2000 and June 2018 was searched and retrieved from online electronic databases. The odds ratio (OR) with its 95% confidence interval (CI) was employed to calculate the effect.Results: A total of 63 case-control studies were finally screened out, including 13,280 asthma patients and 13,340 controls. Eleven SNPs of ADAM33 gene were identified. Our results detected a significant association between ADAM33 T2, Q1, F + 1 and AA genotype of T + 1 polymorphisms and asthma risk in total population. Subgroup analysis by ethnicities showed that the alleles and genotypes of T2, Q1 and F + 1 polymorphisms were associated with asthma susceptibility among Asian populations, while V4 polymorphism was associated with asthma among Caucasian populations. Subgroup analysis by ages showed that T2, F + 1 and ST + 4 polymorphisms were associated with childhood asthma, while Q1 and V4 polymorphisms were associated with asthma risk in adults. Subgroup analysis by asthma severity showed that only the G allele of ADAM33 T1 polymorphism was associated with the severity of asthma when compared with the controls. In addition, T2, Q1 and F + 1 polymorphisms of ADAM33 were significantly associated with increased the asthma risk in Chinese asthma patients.Conclusions: Our results found that T2, Q1 and F + 1 polymorphisms of ADAM33 gene might contribute to asthma risk. Future well-designed case-control studies with large population and more ethnicities are still needed to estimate the association. [ABSTRACT FROM AUTHOR]

The feeding habits of the mud shrimp Solenocera melantho were studied in the East China Sea by analysing 720 stomach contents and comparing diet composition and prey diversity among seasons, sexes and size classes. Feeding activity was linked to season and ovarian condition. The stomach fullness was high during spring and summer and low in fall. Females with advanced ovaries also displayed low stomach fullness, which suggests that feeding activity is affected by the reproductive cycle. The diet of S. melantho consisted of 42 different prey categories, primarily smaller crustaceans, Foraminifera, polychaetes, mollusks and fishes (larvae and eggs). Crustacea,Foraminifera and polychaetes together constituted the dominant prey groups, accounting for 75% of total prey abundance and >50% of total prey occurrence. Diet composition differed among seasons between sexes and size classes. Females and males consumed the same prey items but with different orders of importance. In terms of body size, the small shrimps preferred relatively small food items, such as Foraminifera and algae, whereas larger shrimps preferred polychaetes and crustaceans. Trophic diversity varied among the seasons within both sexes and size classes, with the highest values being observed in fall. Niche overlap was high within both sexes and size classes among seasons (0.6051-0.8740). [ABSTRACT FROM AUTHOR]

Highlights: [ • ] Electron-induced polymer charging is described by contour maps for the first time. [ • ] Leakage current and surface potential are highly correlated in contour features. [ • ] The total charge has the local maximum with respect to sample thickness. [•] Sample thickness normally affects charging effects more than electron mobility. [ABSTRACT FROM AUTHOR]

Abstract: Au nanoparticles dispersed cobalt monoxide (CoO) composite films were fabricated via a chemical solution approach combined with a spin-coating processing. Au particles were spherical approximately and uniformly dispersed in the amorphous CoO matrix. The mean diameters of Au particles in the Au/CoO thin films with 40mol% Au are about 30nm. The optical absorption peaks due to the surface plasmon resonance (SPR) of Au particles were observed in the UV–vis absorption spectra in the wavelength range of 550–650nm. The SPR peaks exhibit a red shift and intensify with increasing Au content from 10 to 40mol%, but show a blue shift and weaken from 40 to 60mol%. The band gap E g decreases with increasing Au contents from 10 to 40mol% but increases by further increasing Au content. [Copyright &y& Elsevier]

Abstract: Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, causes an acute infection of the central nervous system resulting in encephalitis of humans and many kinds of animals. NS5, the largest and most conserved flavivirus protein, is homologous to methyltransferase and RNA-dependent RNA polymerase. RNA interference is an effective anti-viral strategy to inhibit viral replication in vitro. In this study, four short hairpin RNA (shRNA) expression vectors (pS4.1-NS5-201, pS4.1-NS5-455, pS4.1-NS5-699, and pS4.1-NS5-804) targeting the NS5 gene of JEV were employed to target and destroy JEV transcripts. The four shRNAs expression plasmids were individually co-transfected into 293T cells with the plasmid pNS5-EGFP expressing NS5 fused to enhanced green fluorescent protein. The expression level of NS5 was evaluated by fluorescence microscopy, flow cytometry, real time RT-PCR, and Western blot. The four shRNA expression plasmids were also transfected into BHK-21 cells to examine their inhibition of viral replication by indirect immunofluorescence, real time RT-PCR, and Western blot. The results provided strong evidence that shRNAs targeting the NS5 gene could specifically and efficiently inhibit JEV replication. Three out of four plasmids were highly efficient at inhibiting viral replication, including pS4.1-NS5-455, pS4.1-NS5-699, and pS4.1-NS5-804. This was especially true for pS4.1-NS5-699, which reduced the levels of virus RNA and protein the most. Our data suggest that shRNAs could be used as a tool to inhibit JEV replication in vivo. [Copyright &y& Elsevier]

Objective: Obstructive sleep apnea (OSA) is accompanied by widespread abnormal spontaneous regional activity related to cognitive deficits. However, little is known about the topological properties of the functional brain connectome of patients with OSA. This study aimed to use the graph theory approaches to investigate the topological properties and functional connectivity (FC) of the functional connectome in patients with OSA, based on resting-state functional magnetic resonance imaging (rs-fMRI). Methods: Forty-five male patients with newly diagnosed untreated severe OSA and 45 male good sleepers (GSs) underwent a polysomnography (PSG), clinical evaluations, and rs-fMRI scans. The automated anatomical labeling (AAL) atlas was used to construct the functional brain connectome. The topological organization and FC of brain functional networks in patients with OSA were characterized using graph theory methods and investigated the relationship between functional network topology and clinical variables. Results: Both the patients with OSA and the GSs exhibited high-efficiency “small-world” network attributes. However, the patients with OSA exhibited decreased σ, γ, Eglob; increased Lp, λ; and abnormal nodal centralities in several default-mode network (DMN), salience network (SN), and central executive network (CEN) regions. However, the patients with OSA exhibited abnormal functional connections between the DMN, SN, and CEN. The disrupted FC was significantly positive correlations with the global network metrics γ and σ. The global network metrics were significantly correlated with the Epworth Sleepiness Scale (ESS) score, Montreal Cognitive Assessment (MoCA) score, and oxygen desaturation index. Conclusion: The findings suggest that the functional connectome of patients with OSA exhibited disrupted functional integration and segregation, and functional disconnections of the DMN, SN, and CEN. The aberrant topological attributes may be associated with disrupted FC and cognitive functions. These topological abnormalities and disconnections might be potential biomarkers of cognitive impairments in patients with OSA. [ABSTRACT FROM AUTHOR]

Qinghai Lake is a major migratory-bird breeding site that has experienced several highly pathogenic avian influenza virus (AIV) epizootics. Plateau pikas ( Ochotona curzoniae) have previously been implicated in the ecology of avian influenza virus in this region. We first isolated an H9N2 AIV (A/Pika/Menyuan/01/2008) from plateau pikas between November 2008 and October 2009. Sequence analysis showed that the A/Pika/Menyuan/01/2008 AIV was closely related to the H9N2 AIV strain (A/Turkey/Wisconsin/ 1/1966). Our findings suggested that plateau pikas may contribute to AIV epidemiology in the Qinghai Lake region. [ABSTRACT FROM AUTHOR]

Understanding fish's spatial behaviors are important for conservation biology under global warming, as one of the direct responses of fish to climate change is the alterations in distribution. We chose Trichiurus japonicus , which is an ecologically and economically important species in the East China Sea (ECS), to study the vicennial responses of demersal fish to climate change. Eight environmental variables (sea bottom temperature, sea bottom salinity, chlorophyll-a concentration, sea surface height, bathymetry, distance to the coast, slope data, and Terrain ruggedness index) combined with the annual survey data were fitted using integrated nested Laplace approximations and stochastic partial differential equations approach under the Bayesian framework to investigate its distribution shifts in spring during 1999–2019. Based on the distribution model, three spatial behavior indicators (center of gravity, Inertia, and Isotropy) were calculated and correlated with six regional climate indicators (Kuroshio index, Taiwan Warm Current Water index, meridional wind, zonal wind, Pacific Decadal Oscillation, and El Niño/Southern Oscillation) to study the potential cause of its spatial behaviors. During the study periods, the biomass of T. japonicus declined gradually, with a brief increase in 2003, 2007–2008, and 2015–2017. T. japonicus preferred high salinity, warm and deep-water area. The relatively persistent hotspots of T. japonicus in spring were in the offshore waters of eastern Zhejiang Province. The change in the center of gravity during the study periods was not obvious, which implied that the distribution shifts were not necessarily vulnerable for T. japonicus in response to climate change and it has strong adaptability to the environment. We also found that strong Kuroshio Current and Taiwan Warm Current Water as well as cold phased of El Niño/Southern Oscillation were accompanied by weakened spatial aggregation of T. japonicus. Overall, for these types of fish with relatively persistent hotspots, the establishment of seasonally protected areas will contribute to the sustainable utilization of the species. • 20-year-long and almost uninterrupted temporal data were used to develop a distribution model for Trichiurus japonicus in the East China Sea. • T.japonicus preferred high salinity, warm and deep-water area in Spring. • Three spatial indicators showed the spatial behaviors of T.japonicus in spring were mainly affected by wind field, TWC, KI, and ENSO. • T. japonicus has strong adaptability to the environment and its the responses to climate change especially in distribution shifts are not necessarily vulnerable. • Our results have important implications for future fisheries management strategies. [ABSTRACT FROM AUTHOR]

Background:To evaluate the possible relationship between subclinical hypothyroidism (SCH) and metabolic syndrome (MS) and the response to clomiphene citrate (CC) stimulation in women with polycystic ovary syndrome (PCOS). Methods:One hundred and ninety-six women with PCOS were divided into two groups: (1) the SCH group with 92 patients; (2) the euthyroid (EU) group with 104 patients. Physical characteristics and metabolic parameters as well as the reaction to CC stimulating test were compared between these two groups. Results:(1) In the SCH group, significantly higher body mass index, Ferriman–Gallwey score, serum triglyceride, insulin and glucose of oral glucose tolerance test, homeostatic model assessment-insulin resistance (HOMA-IR) and significantly lower serum high-density lipoprotein cholesterol was observed in comparison with those in the EU group (p < 0.05). (2) The prevalence of CC resistance (30.4%), IR (43.5%) and MS (34.8%) in the SCH group was significantly higher than that in the EU group (p< 0.05). Conclusions:SCH was found associated with IR, MS and CC resistance in women with PCOS. PCOS patients with SCH may have a poorer treatment response to ovulation induction with CC. [ABSTRACT FROM PUBLISHER]

[Display omitted] • We developed a 3D lactate electrochemical sensor by layer-by-layer assembly of carbon nanotubes, Prussian blue nanoparticles and lactate oxidase on a 3D scaffold electrode. • Further, the 3D sensor was integrated with collagen hydrogel with C6 glioma cells seeded therein and allowed the successful real-time monitoring of lactate released from C6 glioma cells, providing an effective detection platform for exploring the biochemical response in 3D cell culture systems. Three-dimensional (3D) cell culture has attracted increasing interest in many fields with the significant advantages in reproducing in vivo cell environment. For the characterization of biochemical information, electrochemical sensing has stood out from other techniques in real-time monitoring of living cells, and made an attempt to integrate with hydrogel for investigating the biological processes in 3D cell culture. However, it is still a challenge for real-time monitoring of cellular metabolites therein for the lack of electrochemical activity, and lactate (the product of aerobic glycolysis) is a typical example of such a predicament. Herein, we developed a 3D enzymatic electrochemical sensor that could be integrated with collagen hydrogel to monitor the lactate levels of cells cultured therein. Specifically, lactate oxidase immobilized on the outer surface of 3D sensor could oxidize lactate with H 2 O 2 as a product, and Prussian blue nanoparticles electrodeposited on carbon nanotubes adsorbed electrode conferred the excellent and stable electrocatalytic performance to H 2 O 2 for lactate detection. Further, the 3D sensor was integrated with collagen hydrogel with C6 glioma cells seeded therein and allowed the successful real-time monitoring of lactate released from C6 glioma cells, providing an effective detection platform for exploring the biochemical response in 3D cell culture systems. [ABSTRACT FROM AUTHOR]

Abstract: The development of cell-mediated immunity has been known extremely important in clearing porcine reproductive and respiratory syndrome virus (PRRSV) in infected pigs. However, the PRRS immunology regarding the interaction of T-cells and PRRSV proteins is poorly understood. To identify the T-cell immunodominant epitopes on the membrane (M) protein of PRRSV, a series of 31 overlapping pentadecapeptides covering the entire M protein were designed and synthesized. These peptides were screened by ELIspot analysis for their capabilities to elicit interferon-gamma (IFN-γ) responses in the peripheral blood mononuclear cells (PBMCs), which were collected from pigs immunized with attenuated PRRSV HuN4-F112 strain and challenged with highly pathogenic HuN4 strain. After three rounds of screening, 4 peptides (M3, M6, M8 and M12) were shown to elicit high expression of IFN-γ. The stimulation of high IFN-γ transcription in PBMCs by these 4 peptides was further confirmed in real-time PCR. The sequence alignment revealed that the epitope represented by peptide M6 was fully conserved in all of examined 42 North American genotype II PRRSV isolates and the epitopes represented by peptides M3, M8 and M12 showed 2–4 amino acid replacements. The finding of 4 T-cell immunodominant epitopes in the M protein of PRRSV will be beneficial to the understanding of the development of cell-mediated immunity. [Copyright &y& Elsevier]

Abstract: Pandemic strains of influenza A virus might arise by genetic reassortment between viruses from different hosts. Pigs are susceptible to both human and avian influenza viruses and have been proposed to be intermediate hosts or mixing vessels, for the generation of pandemic influenza viruses through reassortment or adaptation to the mammalian host. In this study, we summarize and report for the first time the coexistence of 10 (A–J) genotypes in pigs in China by analyzing the eight genes of 28 swine H9N2 viruses isolated in China from 1998 to 2007. Swine H9N2 viruses in genotype A and B were completely derived from Y280-like and Shanghai/F/98-like viruses, respectively, which indicated avian-to-pig interspecies transmission of H9N2 viruses did exist in China. The other eight genotype (C–J) viruses might be double-reassortant viruses, in which six genotype (E–J) viruses possessed 1–4 H5-like gene segments indicating they were reassortants of H9 and H5 viruses. In conclusion, genetic diversity of H9N2 influenza viruses from pigs in China provides further evidence that avian to pig interspecies transmission of H9N2 viruses did occur and might result in the generation of new reassortant viruses by genetic reassortment with swine H1N1, H1N2 and H3N2 influenza viruses, therefore, these swine H9N2 influenza viruses might be a potential threat to human health and continuing to carry out swine influenza virus surveillance in China is of great significance. [Copyright &y& Elsevier]

Abstract: Cu/SiO2 nano-composite thin films were prepared by a sol–gel method. The structure and optical property of the films were investigated with a special emphasis on the influences of Cu content. Cu particles were basically spherical and dispersed in the SiO2 matrix. The optical absorption peaks due to the surface plasmon resonance of Cu particles were observed in the wavelength range of 550–600nm. The absorption property is enhanced with increasing Cu content, showing a maximum value in the films with 30at.% Cu. Increasing Cu content above 30at.% results in a decrease in absorption intensity. The absorption peak shows a red-shift trend with increasing Cu content from 5 to 30at.% and then turns to blue-shift by further increasing the Cu content from 30 to 35at.%. The band gap Eg decreases with increasing Cu content from 10 to 30at.% but increases by further increasing Cu content. [Copyright &y& Elsevier]

Abstract: DNA and recombinant virus vaccines against swine influenza virus (SIV) have been pursued with promising results, but induce poor immunogenicity. This study evaluated the effects of a vaccine regimen in mice including priming with three DNA vaccines expressing soluble HA (sHA), complete HA (tmHA), or sHA fused with three copies murine C3d (sHA–mC3d3) and boosting with recombinant pseudorabies virus expressing HA (rPRV–HA). Immune responses were monitored by ELISA, HI assays, and virus neutralization. Protective efficacy was evaluated by virus isolation from lungs, distribution in tissues, and pathology following challenge with H3N2 SIV. Priming with sHA–mC3d3 and boosting with rPRV–HA induced higher levels of HA-specific antibodies and yielded the most effective protection. This finding implied that priming with a DNA vaccine expressing C3d fused with antigen and boosting with a recombinant vector vaccine is an effective way to induce protective humoral immunity and prevent some infectious diseases. [Copyright &y& Elsevier]

Abstract: In vitro and in vivo studies have demonstrated that inhibition of phospholamban (PLB) expression in myocardium can restore left ventricular systolic function in failing heart. Ultrasound mediated microbubble destruction provides a new option for noninvasive gene transfer in heart. In this study, we transfered pAAV-antisense phospholamban (pAAV-asPLB) to the hearts of myocardial infarction (MI) mice, using ultrasound mediated microbubble destruction. Then we estimated the protein levels of PLB, Ser16-PLB and cardiac sarcoplasmic reticulum Ca2+ ATPase (SERCA). The left ventricular ejection fraction (LVEF), fraction shortening (FS) and SERCA activity were measured as well. MI mice were generated by ligating the left anterior descending coronary artery. Microbubbles were prepared by sonicated perfluorocarbon gas with dextrose and albumin. A mixture of pAAV-asPLB plasmid and microbubble was injected via tail vein while the heart was simultaneously exposed to ultrasound via transthoracic insonation. Three weeks later, LVEF (48.2±5.18% vs 39.1±5.38%, p <0.05), FS (19.6±2.59% vs 16.0±2.29%, p <0.05), SERCA activity (3.00±0.29 vs 2.12±0.30, p <0.05) and Ser16-PLB protein level (0.8±0.25 vs 0.46±0.18, p <0.05) were increased while PLB protein level (1.45±0.38 vs 2.05±0.31, p <0.05) was decreased compared with the MI mice with saline injection. The above parameters in MI mice with only pAAV-asPLB plasmid injection or pAAV-asPLB plasmid combined with ultrasound alone were not significantly improved. pAAV-LacZ was used as a reporter gene to determine the efficiency and localization of transfection. The expression of β-galactosidase was not found in liver, lung and brain, but found only in tubular epithelial cells of kidney and found in heart. These results confirm that asPLB gene transfection can be achieved by ultrasound mediated microbubble destruction with organ specificity. The effective transfection can partly restore heart function in MI mice. (E-mail: s0hu0001@hotmail.com) [Copyright &y& Elsevier]

Abstract: Since April 2006, swine herds have experienced the outbreaks of a highly pathogenic porcine reproductive and respiratory syndrome (PRRS) in China. To explore the possible mechanism of the emergence of the highly pathogenic PRRS and more fully understand the extent of genetic diversity of PRRSV in China, we analyzed the ORF5 gene sequences of 159 representative PRRSV isolates in 16 provinces from 2006 to 2008. Sequence and phylogenetic analyses showed that all these 159 isolates belonged to the North American genotype and were further divided into six subgenotypes; 140 of 159 isolates were closely related to the highly pathogenic PRRSV with 98.5–100% nucleotide and 98.3–100% amino acid sequence identities and belonged to Subgenotype I; and 3, 8, 4, 3, 1 of 159 isolates were part of Subgenotypes II–VI, respectively. Amino acid analysis of the GP5 protein revealed that all the isolates in Subgenotypes I–III were found to be highly variable in the primary neutralizing epitope; most of the isolates in Subgenotypes I and IV had more glycosylation sites than those in Subgenotypes II, III, V and VI; and 1, 5, and 9 unique amino acid mutations were observed in Subgenotypes I, IV and VI, respectively. In conclusion, our study provides the evidence of coexistence of six different subgenotype isolates in pigs in China from 2006 to 2008, and emphasizes the importance of reinforcing PRRSV surveillance, especially after the emergence of highly pathogenic PRRS in China. [Copyright &y& Elsevier]

Abstract: As pigs are susceptible to both human and avian influenza viruses, they have been proposed to be intermediate hosts or mixing vessels for the generation of pandemic influenza viruses through reassortment or adaptation to the mammalian host. In this study, we reported avian-like H1N1 and novel ressortant H1N2 influenza viruses from pigs in China. Homology and phylogenetic analyses showed that the H1N1 virus (A/swine/Zhejiang/1/07) was closely to avian-like H1N1 viruses and seemed to be derived from the European swine H1N1 viruses, which was for the first time reported in China; and the two H1N2 viruses (A/swine/Shanghai/1/07 and A/swine/Guangxi/13/06) were novel ressortant H1N2 influenza viruses containing genes from the classical swine (HA, NP, M and NS), human (NA and PB1) and avian (PB2 and PA) lineages, which indicted that the reassortment among human, avian, and swine influenza viruses had taken place in pigs in China and resulted in the generation of new viruses. The isolation of avian-like H1N1 influenza virus originated from the European swine H1N1 viruses, especially the emergence of two novel ressortant H1N2 influenza viruses provides further evidence that pigs serve as intermediate hosts or “mixing vessels”, and swine influenza virus surveillance in China should be given a high priority. [Copyright &y& Elsevier]

Abstract: Urotensin II (U-II), a cyclic undecapeptide, is the natural ligand of the urotensin II (UT) receptor, a G protein-coupled receptor. In the present study, we used the substituted-cysteine accessibility method to identify specific residues in transmembrane domains (TMDs) six and seven of the rat urotensin II receptor (rUT) that contribute to the formation of the binding pocket of the receptor. Each residue in the R256(6.32)-Q283(6.59) fragment of TMD6 and the A295(7.31)-T321(7.57) fragment of TMD7 was mutated, individually, to a cysteine. The resulting mutants were expressed in COS-7 cells, which were subsequently treated with the positively charged methanethiosulfonate-ethylammonium (MTSEA) or the negatively charged methanethiosulfonate-ethylsulfonate (MTSES) sulfhydryl-specific alkylating agents. MTSEA treatment resulted in a significant reduction in the binding of TMD6 mutants F268C(6.44) and W278C(6.54) and TMD7 mutants L298C(7.34), T302C(7.38), and T303C(7.39) to 125I-U-II. MTSES treatment resulted in a significant reduction in the binding of two additional mutants, namely L282C(6.58) in TMD6 and Y300C(7.36) in TMD7. These results suggest that specific residues orient themselves within the water-accessible binding pocket of the rUT receptor. This approach, which allowed us to identify key determinants in TMD6 and TMD7 that contribute to the UT receptor binding pocket, enabled us to further refine our homology-based model of how U-II interacts with its cognate receptor. [Copyright &y& Elsevier]

Abstract: Classical swine and avian-like H1N1 influenza viruses were reported widely in swine population worldwide, but human-like H1N1 swine viruses were reported occasionally. In 2006, a human-like H1N1 swine virus (A/swine/Guangdong/96/06) was isolated from pigs in Guangdong province, which was reported in China for the first time. To get further evidence for infection of pigs with human-like H1N1 influenza viruses, we analyzed eight gene segments of three human-like swine H1N1 viruses (A/swine/Guangdong/96/06, A/swine/Tianjin/01/04 and A/swine/Henan/01/06) isolated in China. All the eight genes of the three viruses are highly homologous to recent (about 2000) and early (1980s) human H1N1 influenza viruses, respectively. Phylogenetic analyses revealed that A/Swine/Guangdong/96/06 was directly derived from about 2000 human H1N1 influenza viruses, while A/swine/Tianjin/01/04 and A/swine/Henan/01/06 seemed to be descendants of human H1N1 viruses circulating in 1980s. Seroprevalence of our isolate (A/swine/Guangdong/96/06) confirmed the presence of human-like H1N1 virus in pigs in China. Existence of these influenza viruses, especially older viruses (A/swine/Tianjin/01/04 and A/swine/Henan/01/06), indicates that human-like H1N1 influenza viruses may remain invariant for long periods in pigs and provides the evidence that pigs serve as reservoirs of older influenza viruses for human pandemics. [Copyright &y& Elsevier]