Recombinant pseudorabies virus with gI/gE deletion generated by overlapping polymerase chain reaction and homologous recombination technology induces protection against the PRV variant PRV-GD2013.

BACKGROUND: Since 2011, numerous highly virulent and antigenic variant viral strains have been reported in pigs that were vaccinated against the swine pseudorabies virus. These infections have led to substantial economic losses in the Chinese swine industry. RESULTS: This study, constructed a novel recombinant vaccine strain with gI/gE deletion (PRV-GD2013-ΔgI/gE) by overlapping PCR and homologous recombination technology. The growth curves and plaque morphology of the recombinant virus were similar to those of the parental strain. However, PRV-GD2013-ΔgI/gE infection was significantly attenuated in mice compared with that of PRV-GD2013. Two-week-old piglets had normal rectal temperatures and displayed no clinical symptoms after being inoculated with 105 TCID50 PRV-GD2013-ΔgI/gE, indicating that the recombinant virus was avirulent in piglets. Piglets were immunized with different doses of PRV-GD2013-ΔgI/gE, or a single dose of Bartha-K61 or DMEM, and infected with PRV-GD2013 at 14 days post-vaccination. Piglets given high doses of PRV-GD2013-ΔgI/gE showed no obvious clinical symptoms, and their antibody levels were higher than those of other groups, indicating that the piglets were completely protected from PRV-GD2013. CONCLUSIONS: The PRV-GD2013-ΔgI/gE vaccine strain could be effective for immunizing Chinese swine herds against the pseudorabies virus (PRV) strain.

Induction of autophagy and suppression of type I IFN secretion by CSFV.

Macroautophagy/autophagy plays an essential role in cellular responses to pathogens. However, the precise mechanisms and signaling pathways that modulate cellular autophagy in classical swine fever virus (CSFV)-infected host cells have not been confirmed. In this study, we showed that CSFV infection inhibits the phosphorylation of MTOR (mechanistic target of rapamycin kinase), subsequently leading to autophagy initiation. We also show that MAPK/ERK (mitogen-activated protein kinase) signaling is involved in CSFV-induced autophagy. The CSFV-induced inhibition of AKT/PKB (AKT serine/threonine kinase)-MTOR was observed to be partially responsible for the MTOR inactivation and subsequent autophagy initiation. Moreover, the CAMKK2/CaMKKß (calcium/calmodulin dependent protein kinase kinase 2)-PRKAA/AMPK (protein kinase AMP-activated catalytic subunit alpha) axis was found to be involved in CSFV-induced autophagy. Meanwhile, CSFV non-structural protein NS5A induced autophagy via the CAMKK2-PRKAA-MTOR signaling pathway but not the AKT-MTOR or MAPK1/ERK2-MAPK3/ERK1-MTOR pathway. Although the AKT-MTOR pathway also plays an important role in the induction of autophagy by CSFV. We also found the interaction between HSP90AB1/HSPCB and NS5A by tandem affinity purification/liquid chromatography-mass spectrometry (LC-MS) and immunoprecipitation. Furthermore, the CSFV-induced [Ca2+]cyto increase potently induced autophagy through CAMKK2 and PRKAA. Moreover, we isolated and identified the BECN1/Beclin 1 protein complexes by tandem affinity purification/LC-MS and immunoprecipitation, the interaction between BECN1 and MAVS was confirmed by immunoprecipitation, laser scanning confocal microscope technology, and GST affinity-isolation experiments. Furthermore, CSFV-mediated autophagy suppressing type I IFN production is related to the interaction between MAVS and BECN1. Finally, the modulation of autophagy induction pathways by different autophagy regulatory factors significantly affected the replication of CSFV.Abbreviations: AKT: AKT serine/threonine kinase; AMPK: Adenosine monophosphate-activated protein kinase; CAMKK2: Calcium/calmodulin dependent protein kinase kinase 2; CSFV: Classical swine fever virus; HRP: Horseradish peroxidase; HSP90AB1: Heat shock protein 90 alpha family class B member 1; IFN: Interferon; ISGs: IFN-stimulated genes; LC-MS: Liquid chromatography-mass spectrometry; MAP1LC3/LC3: Microtubule associated protein 1 light chain 3; MAPK: Mitogen-activated protein kinase; MAVS: Mitochondrial antiviral signaling protein; MOI: Multiplicity of infection; MTOR: Mechanistic target of rapamycin kinase; PBS: Phosphate-buffered saline; PRKAA: Protein kinase AMP-activated catalytic subunit alpha; shRNA: short hairpin RNA.

LncRNA OIP5-AS1 reduces α-synuclein aggregation and toxicity by targeting miR-126 to activate PLK2 in human neuroblastoma SH-SY5Y cells.

It has been reported that many long noncoding RNAs (lncRNA) are abnormally expressed in Parkinson's disease (PD). However, the knowledge about the role of dysregulated lncRNA in the pathological process of PD and the potential molecular regulation mechanism is still limited. Our immunofluorescence data show that miR-126 enhances the aggregation and toxicity of synuclein, while lncRNA OIP5-AS1 reduces the aggregation and toxicity of MPP + induced α-synuclein by targeting miR-126. Luciferase experiments have found that miR-126 regulates α-synuclein by targeting PLK2. Western blot and IP experimental analysis showed that this process is achieved by regulating PLK2/α-synuclein autophagy. In conclusion, our data indicate that OIP5-AS1 promotes the autophagy of PLK2-α-synuclein by targeting the miR-126 axis with pathogenic factors, thus reducing the aggregation toxicity of α-synuclein, which It will help better to understand the mechanism of dopaminergic neuron loss in PD and provide novel treatment options.

Dietary and serum selenium in coronary heart disease and all-cause mortality: An international perspective.

BACKGROUND AND OBJECTIVES: The objective of this study was to explore the associations of dietary selenium and serum selenium concentration with coronary heart disease (CHD) prevalence and all-cause mortality among participants in United States. METHODS AND STUDY DESIGN: Using data collected from the National Health and Nutrition Examination Survey (NHANES) 1999-2006, 17867 individuals were included. Logistic regression analyses were used to explore the associations between dietary selenium intake and serum selenium concentration and prevalent of CHD. Multivariable Cox regression was used to identify the association between dietary selenium intake and all-cause mortality. The nonlinear relationships were assessed using generalized additive models. RESULTS: A U-shaped association between dietary intake of selenium and all-cause mortality was observed. Compared with the lowest quartile, the second quartile of dietary intake of selenium was inversely associated with all-cause mortality (Hazard ratio [HR]: 0.802, 95% confidence interval [CI]: 0.658, 0.977, p=0.029). There was no evidence of association between dietary selenium intake and CHD risk (Odds ratio [OR]: 1.001, 95% CI: 0.999, 1.003, p=0.206). Furthermore, serum selenium concentration was negatively associated with CHD risk (OR: 0.989, 95% CI: 0.981, 0.997, p=0.006). Comparing with the lowest quartile, participants with the highest serum selenium concentration had a statistically significant decreased prevalence of CHD, with OR (95% CI) of 0.417 (0.259, 0.669) (p<0.001). The smoothing curve also showed a non-linear relationship between serum selenium and risk of CHD. CONCLUSIONS: This analysis suggested that a higher serum selenium concentration was associated with reduced risk of CHD, and that the relationship was non-linear. In addition, an appropriate dietary selenium intake might reduce all-cause mortality.

Association between daytime napping duration and depression in middle-aged and elderly Chinese: evidence from the China Health and Retirement Longitudinal Study (CHARLS): A cross-sectional study in China.

The effect of the afternoon napping duration on the risk of depression has not been well established, particularly with regard to sex and age differences. The present study examines the association between afternoon napping duration and depression stratified by sex and age among Chinese adults aged 45 years or older.The 2011 to 2012 survey of the China Health and Retirement Longitudinal Study was utilized, including 5746 participants. We conducted logistic regression with the overall sample and subjects stratified by sex and age.Elderly men with short napping (<30 minutes) had lower odds of having depression symptoms compared with those with no napping group (OR = 0.66, 95% CI = 0.44-0.97). In addition, the finding indicated that middle-aged women with long napping (≥90 min) had a marginally significant difference than those in reference, which showed a negative effect on depression (OR = 0.72, 95% CI = 0.51-1.01).Our findings revealed that extended daytime napping duration can decrease the risk of depression status among middle and elderly people. Moreover, relevant promotion measures should be adopted, such as a suitable rest environment and regular napping habits. The potential mechanism should be clarified by a longitudinal survey to examine the specific causality.

Activation of Interleukin-1ß Release by the Classical Swine Fever Virus Is Dependent on the NLRP3 Inflammasome, Which Affects Virus Growth in Monocytes.

Classical swine fever virus (CSFV) is a classic Flavivirus that causes the acute, febrile, and highly contagious disease known as classical swine fever (CSF). Inflammasomes are molecular platforms that trigger the maturation of proinflammatory cytokines to engage innate immune defenses that are induced upon cellular infection or stress. However, the relationship between the inflammasome and CSFV infection has not been thoroughly characterized. To understand the function of the inflammasome response to CSFV infection, we infected porcine peripheral blood monocytes (PBMCs) with CSFV. Our results indicated that CSFV infection induced both the generation of pro-interleukin-1ß (pro-IL-1ß) and its processing in monocytes, leading to the maturation and secretion of IL-1ß through the activation of caspase 1. Moreover, CSFV infection in PBMCs induced the production and cleavage of gasdermin D (GSDMD), which is an inducer of pyroptosis. Additional studies showed that CSFV-induced IL-1ß secretion was mediated by NLRP3 and that CSFV infection could sufficiently activate the assembly of the NLRP3 inflammasome in monocytes. These results revealed that CSFV infection inhibited the expression of NLRP3, and knockdown of NLRP3 enhanced the replication of CSFV. In conclusion, these findings demonstrate that the NLRP3 inflammasome plays an important role in the innate immune response to CSFV infection.

CSFV Infection Up-Regulates the Unfolded Protein Response to Promote Its Replication.

Classical swine fever (CSF) is an OIE-listed, highly contagious animal disease caused by classical swine fever virus (CSFV). The endoplasmic reticulum (ER) is an organelle in which the replication of many RNA viruses takes place. During viral infection, a series of events elicited in cells can destroy the ER homeostasis that cause ER stress and induce an unfolded protein response (UPR). In this study, we demonstrate that ER stress was induced during CSFV infection as several UPR-responsive elements such as XBP1(s), GRP78 and CHOP were up-regulated. Specifically, CSFV transiently activated IRE1 pathway at the initial stage of infection but rapidly switched off, likely due to the reduction in cytoplasm Ca2+ after viral incubation. Additionally, our data show that the ER stress induced by CSFV can promote CSFV production, which the IRE1 pathway play an important role in it. Evidence of ER stress in vivo was also confirmed by the marked elevation of GRP78 in CSFV-infected pig PBMC and tissues. Collectively, these data indicate that the ER stress was induced upon CSFV infection and that the activation of the IRE1 pathway benefits CSFV replication.

Effect of plant growth-promoting bacteria (PGPR) and arbuscular mycorrhizal fungi (AMF) inoculation on oats in saline-alkali soil contaminated by petroleum to enhance phytoremediation.

To investigate the effect of plant growth-promoting bacteria (PGPR) and arbuscular mycorrhizal fungi (AMF) on phytoremediation in saline-alkali soil contaminated by petroleum, saline-alkali soil samples were artificially mixed with different amount of oil, 5 and 10 g/kg, respectively. Pot experiments with oat plants (Avena sativa) were conducted under greenhouse condition for 60 days. Plant biomass, physiological parameters in leaves, soil enzymes, and degradation rate of total petroleum hydrocarbon were measured. The result demonstrated that petroleum inhibited the growth of the plant; however, inoculation with PGPR in combination with AMF resulted in an increase in dry weight and stem height compared with noninoculated controls. Petroleum stress increased the accumulation of malondialdehyde (MDA) and free proline and the activities of the antioxidant enzyme such as superoxide dismutase, catalase, and peroxidase. Application of PGPR and AMF augmented the activities of three enzymes compared to their respective uninoculated controls, but decreased the MDA and free proline contents, indicating that PGPR and AMF could make the plants more tolerant to harmful hydrocarbon contaminants. It also improved the soil quality by increasing the activities of soil enzyme such as urease, sucrase, and dehydrogenase. In addition, the degradation rate of total petroleum hydrocarbon during treatment with PGPR and AMF in moderately contaminated soil reached a maximum of 49.73%. Therefore, we concluded the plants treated with a combination of PGPR and AMF had a high potential to contribute to remediation of saline-alkali soil contaminated with petroleum.

Contribution of reactive oxygen species (ROS) to genotoxicity of nitrobenzene on V. faba.

Nitrobenzene is an important organic intermediate widely used in industry that can be hazardous to the environment. In our previous study, nitrobenzene showed genotoxic effect on soybean and tobacco plants at concentrations in the culture medium higher than 10 mg/L. The genotoxicity of nitrobenzene has been hypothesized to be multifactorial and reflective of the generation of free radicals; however, the mechanism has not been fully elucidated. The aim of this study was to investigate the relationship between the induction of genotoxicity and the production of free radicals in young seedlings of V. faba exposed to nitrobenzene, nitrobenzene + Vitamin C, and the controls (distilled water or Vitamin C). Micronucleus and chromosome aberration assays performed on root and leaf tissue of V. faba seedlings exposed to nitrobenzene (25 mg/L) demonstrated genotoxic effects which were partly reduced by Vitamin C at 25 mg/L. Increases in lipid peroxidase, O2•-, H2O2, superoxide dismutase and catalase activities were also observed in these tissues along with an attenuation of their induction by Vitamin C. Concomitant occurrence of genotoxicity and the generation of free radicals that are attenuated in the presence of Vitamin C, a scavenger of cellular free radicals, indicate that reactive oxygen species may contributes to genotoxicity of nitrobenzene in V. faba. These results are valuable for further understanding the genotoxicity mechanism of nitrobenzene.