Identification and integrated analysis of lncRNAs and miRNAs in IPEC-J2 cells provide novel insight into the regulation of the innate immune response by PDCoV infection.

BACKGROUND: Noncoding RNAs (ncRNAs), including microRNAs (miRNAs) and long noncoding RNAs (lncRNAs), are pivotal regulators involved in the pathogenic mechanism of multiple coronaviruses. Porcine deltacoronavirus (PDCoV) has evolved multiple strategies to escape the innate immune response of host cells, but whether ncRNAs are involved in this process during PDCoV infection is still unknown. RESULTS: In this study, the expression profiles of miRNAs, lncRNAs and mRNAs in IPEC-J2 cells infected with PDCoV at 0, 12 and 24 hours postinfection (hpi) were identified through small RNA and RNA sequencing. The differentially expressed miRNAs (DEmiRNAs), lncRNAs (DElncRNAs) and mRNAs (DEmRNAs) were screened from the comparison group of IPEC-J2 cells at 0 and 12 hpi as well as the comparison group of IPEC-J2 cells at 12 and 24 hpi. The target genes of these DEncRNAs were predicted. The bioinformatics analysis of the target genes revealed multiple significantly enriched functions and pathways. Among them, the genes that were associated with innate immunity were specifically screened. The expression of innate immunity-related ncRNAs and mRNAs was validated by RT-qPCR. Competing endogenous RNA (ceRNA) regulatory networks among innate immunity-related ncRNAs and their target mRNAs were established. Moreover, we found that the replication of PDCoV was significantly inhibited by two innate immunity-related miRNAs, ssc-miR-30c-3p and ssc-miR-374b-3p, in IPEC-J2 cells. CONCLUSIONS: This study provides a data platform to conduct studies of the pathogenic mechanism of PDCoV from a new perspective and will be helpful for further elucidation of the functional role of ncRNAs involved in PDCoV escaping the innate immune response.

Revealing the mechanism of post-harvest processing on rose quality based on dynamic changes in water content, enzyme activity, volatile and non-volatile metabolites.

Existing studies on post-harvest processing of edible roses have mainly focused on processing techniques and physicochemical properties of the final dried products, with limited studies on how changes in metabolites during processing affect the quality of these products. This study investigated changes in water content and status, enzyme activity, phenolic compounds, and volatile and non-volatile compounds during processing and revealed the mechanisms by which post-harvest processing (drying without blanching (WBD) and drying with blanching (BD)) affects the quality of dried roses by establishing their correlations. Results showed that the blanching reduced the relative content of free water and water activity, thus reducing the subsequent drying time and enzyme activity. The BD method caused higher levels of phenolic compounds than the WBD method in terms of gallic acid, ellagic acid, epicatechin, and quercetin. The OPLS-DA analysis identified 6 differential volatiles out of 72 detected volatiles, contributing to the unique aroma of dried roses by activating olfactory receptors through hydrogen bonding and hydrophobic interactions. 58 differential metabolites were screened from 964 non-volatile metabolites. KEGG pathway analysis revealed that the changes in volatile and non-volatile metabolites induced by different processing methods were due to the effect of blanching on glutathione and fatty acid metabolism. These findings provide a comprehensive understanding of how post-harvest processing affects the quality of dried roses.

Modulation of the properties of starch gels by a one-step extrusion modification method based on Ca2+-citric acid synergistic crosslinking.

Citric acid (CA) is a green and safe food-grade crosslinking agent for starch, but its high crosslinking temperature limits its application. In this study, a "one-step" extrusion modification method based on Ca2+-esterification synergistic crosslinking was proposed for the preparation of high gel performance crosslinked starch at low temperatures (90 °C). The linear and nonlinear rheological properties of crosslinked starch were comprehensively characterized, and the enhancement effect of synergistic crosslinking reactions on starch gel properties was quantitatively studied. The results show that the elastic modulus of the synergistically crosslinked starch (SC-0.5Ca2+, G' = 3116 ± 36) was significantly increased by 879 % compared to the elastic modulus of starch without synergistically crosslinked modification (SC, G' = 318 ± 9). The elastic modulus of starch gels can be adjusted by changing the ion concentration. Nonlinear rheological Lissajous curve analysis results show that the synergistic crosslinked gel system has a stronger anti-deformation ability. In addition, the honeycomb porous structure and smaller pore size distribution of the synergistic crosslinked gels were characterized using scanning SEM. The XPS, FTIR and XRD results suggest that the synergistic crosslinking enhancement effect may involve various molecular forces such as electrostatic attraction, hydrogen bonding and ester bonding.

Enhancement of Starch Gel Properties Using Ionic Synergistic Multiple Crosslinking Extrusion Modification.

Crosslinking is a promising method to modulate the gel properties of food-grade starch gels. Still, the poor crosslinking effect of a single type of crosslinker limits the application of this method in starch gel modification. In this study, an Ca2+ synergistic multiple crosslinking modification method was proposed to prepare crosslinked starches with good gel properties and setting. The rheological properties of the samples were tested. The modified sample (SC-Ca-N3, G' = 1347 ± 27) showed a 79% improvement compared to the starch without synergistic crosslinking modification (SC-N, G' = 752 ± 6). The elastic modulus of starch gels can be adjusted by changing the degree of the crosslinking reaction. The results of nonlinear rheological Lissajous curve analysis showed that the synergistically crosslinked gel system strongly resisted deformation. In addition, the microstructure of the modified samples was characterized using scanning electron microscopy. The XPS, FTIR, and XRD results indicated that multiple molecular forces participate in the synergistic crosslinking reaction.

Isolation, identification and genetic characterization analysis of a fowl aviadenovirus serotype 4 strain from Tianjin, China.

A fowl aviadenovirus serotype 4 (FAdV-4), Y17215-1, was isolated from the liver of chickens with Hydropericardium-hepatitissyndrome(HHS) in a chicken farm of Tianjin, China. Obvious cytopathic effects were observed in the infected chicken liver hepatocellular carcinoma cell line (LMH cells) at 24 h post infection (hpi), which consisted of enlarger and rounder shape of cells. The typical and specific green fluorescence was observed by indirect immunofluorescence assay (IFA). Tissue Culture Infectious Dose50 (TCID50) of it measured after five stable passage in LMH cells reached 106.5TCID50/0.1 mL. The strain was inoculated through allantoic membrane of 10-day specific pathogen free(SPF) Chick embryos, the thicker allantoic membranes were observed at 120 hpi. 7-day-old SPF chickens were inoculated with the strain via intramuscular (i.m.) or intranasal (i.n.) injection which resulted in 100% mortality of test chickens. Additionally, the sickness and death of cohabitation chickens in the test group were observed which indicated that the virus can infect healthy chickens by horizontal transmission. The sick chickens showed depression, anorexia and diarrhea with green watery feces. Y17215-1-inoculated chickens mainly presented swollen liver with blood spot, and the enhancement of effusion or yellow gel like effusion that were observed in the pericardium through necropsy. Histopathological examination showed focal necrosis of hepatocytes and characteristic eosinophilic inclusion bodies in the cytoplasm. The results showed that the Y17215-1 isolate had high pathogenicity to SPF chickens. The phylogenetic analysis of the major structural proteins including hexon, fiber-1 and fiber-2 revealed that Y17215-1 strain belongs to C species of fowl aviadenovirus of aviadenovirus family, and has high homology with other Chinese strains isolated in recent years, but was distinct from ON1、MX-SHP95、KR5 and other foreign isolates. This study laid a foundation for further study of epidemiological investigation, pathogenic mechanism as well as the diagnosis and control technology of FAdV-4.

Epidemiology of Porcine Pseudorabies from 2010 to 2018 in Tianjin, China.

Pseudorabies (PR), the causative agent of Aujeszky's disease, has rapidly increased in recent years and has caused significant economic losses. To understand the seroprevalence and epidemiological characteristics of PR in Tianjin, China, a total of 23,627 blood and 1,093 tissue samples were collected from 228 pig farms during January 2010 to December 2018. The Pseudorabies virus (PRV) glycoprotein E (gE) antibody was tested by enzyme-linked immunosorbent assay (ELISA), and wild-type PRV (WT PRV) was detected by gE-polymerase chain reaction (PCR). Macroscopic and microscopic lesions were observed in tissue samples. The results showed that 46.70% of the serum samples and 49.76% of pig farms were seropositive for PRV gE antibody based on the ELISA results, and 13.54% of the tissue samples were positive for WT PRV detected by PCR. The positive rate of serum samples increased rapidly after 2011 and reached 62.40% in 2013. Although it gradually decreased from 2014 to 2018, the positive rate of serum samples remained at a high level. The positive rate of pig farms showed the same trend. Moreover, after 2011, the detection rate of WT PRV was increased rapidly and was significantly higher than in 2010 and 2011. Macroscopic and microscopic lesions were observed in various tissues during histopathological examination. Based on univariate analysis, the increased risk of seropositivity was associated with the immune status and infection in sows and fattening pigs. These findings demonstrate that PR was prevalent in the region of Tianjin, China. These epidemiological data can assist in the control of PR.

Efficacy evaluation of two commercial modified-live virus vaccines against a novel recombinant type 2 porcine reproductive and respiratory syndrome virus.

NADC30-like porcine reproductive and respiratory syndrome virus (PRRSV) causing clinical disease outbreaks has been recently reported in China. The recombination occurring among PRRSV strains could lead to the emergence of novel and more virulent viruses. In our previous study, a novel recombinant type 2 PRRSV (TJnh1501) between NADC30-like and modified-live virus (MLV)-like derived from the Chinese highly pathogenic PRRSV was shown to have higher pathogenicity than NADC30-like PRRSV. It remains unknown whether the emergence of the novel recombinant PRRSV strain can lead to variable protection efficacy of the MLV vaccines. In this paper, two typical commercial MLV vaccines were used to evaluate their efficacy to block TJnh1501 infection and onset of clinical symptoms. Our results showed that both MLV vaccines could shorten the period of fever and reduce viral loads in sera, but were not able to reduce the clinical signs and lung lesions indicating that the two commercial MLV vaccines provide limited cross-protection efficacy against the novel recombinant type 2 PRRSV infection. This study gives valuable suggestions for the use of MLV vaccines to control PRRSV infection in the field.

Novel triple-reassortant H1N1 swine influenza viruses in pigs in Tianjin, Northern China.

Pigs are susceptible to both human and avian influenza viruses and therefore have been proposed to be mixing vessels for the generation of pandemic influenza viruses through reassortment. In this study, for the first time, we report the isolation and genetic analyses of three novel triple-reassortant H1N1 swine influenza viruses from pigs in Tianjin, Northern China. Phylogenetic analysis showed that these novel viruses contained genes from the 2009 pandemic H1N1 (PB2, PB1, PA and NP), Eurasian swine (HA, NA and M) and triple-reassortant swine (NS) lineages. This indicated that the reassortment among the 2009 pandemic H1N1, Eurasian swine and triple-reassortant swine influenza viruses had taken place in pigs in Tianjin and resulted in the generation of new viruses. Furthermore, three human-like H1N1, two classical swine H1N1 and two Eurasian swine H1N1 viruses were also isolated during the swine influenza virus surveillance from 2009 to 2013, which indicated that multiple genetic lineages of swine H1N1 viruses were co-circulating in the swine population in Tianjin, China. The emergence of novel triple-reassortant H1N1 swine influenza viruses may be a potential threat to human health and emphasizes the importance of further continuous surveillance.