Pseudorabies virus exploits N6-methyladenosine modification to promote viral replication.

Introduction: Pseudorabies virus (PRV) is the pathogenic virus of porcine pseudorabies (PR), belonging to the Herpesviridae family. PRV has a wide range of hosts and in recent years has also been reported to infect humans. N6-methyladenosine (m6A) modification is the major pathway of RNA post-transcriptional modification. Whether m6A modification participates in the regulation of PRV replication is unknown. Methods: Here, we investigated that the m6A modification was abundant in the PRV transcripts and PRV infection affected the epitranscriptome of host cells. Knockdown of cellular m6A methyltransferases METTL3 and METTL14 and the specific binding proteins YTHDF2 and YTHDF3 inhibited PRV replication, while silencing of demethylase ALKBH5 promoted PRV output. The overexpression of METTL14 induced more efficient virus proliferation in PRV-infected PK15 cells. Inhibition of m6A modification by 3-deazaadenosine (3-DAA), a m6A modification inhibitor, could significantly reduce viral replication. Results and Discussion: Taken together, m6A modification played a positive role in the regulation of PRV replication and gene expression. Our research revealed m6A modification sites in PRV transcripts and determined that m6A modification dynamically mediated the interaction between PRV and host.

Effect of anemoside B4 on milk whey in clinical mastitis-affected cows elucidated using tandem mass tag (TMT)-based quantitative proteomics.

Intramuscular injection of anemoside B4 (AB4) has a superior therapeutic effect on clinical mastitis in lactating cows. Here, we explored AB4's effect on milk whey in clinical mastitis-affected cows using proteomics. Among fifty clinical mastitis cows received AB4 administration (0.05 ml/kg/day, for 7 days), twelve healed cows were selected and marked as group T. Twelve clinically heathy cows received the same dose of saline for 7 days, marked as group C. Collected milk whey of group T before and after AB4 administration marked as T1 and T2, respectively. The milk whey of group C after saline injection marked as C1. Milk whey protein changes were detected using tandem mass tag-based quantitative proteomic. We identified 872 quantifiable proteins in the samples. Among them, 511 proteins between T1 and C1, and 361 proteins between T2 and T1 were significantly altered. T1 than C1 had significantly more proteins associated with inflammatory damage and trans-endothelial migration of leukocytes, whereas these proteins were reduced in T2 treated with AB4. Compared with C, proteins associated with fibrin clot degradation and complement system activation were downregulated in T1 but upregulated in T2. In summary, AB4 can exert its therapeutic effect on clinical mastitis in cows mainly by reducing inflammatory damage, activating the complement system, inhibiting trans-endothelial migration of leukocytes, and promoting degradation of milk fibrin clots.

Antioxidant Capacity and Protective Effect of Cow Placenta Extract on D-Galactose-Induced Skin Aging in Mice.

Placental extract has been used for skin care and delaying skin aging. Cow placenta is an abundant resource with a large mass, which has not been harnessed effectively. Cow placenta extract (CPE) has the functions of antioxidation, anti-inflammatory, promoting growth and development, and promoting hair growth. However, little is known about the effect of oral administration of cow placenta extract on skin conditions. Therefore, the present study aimed to investigate the antioxidant capacity of CPE in vitro and in vivo and its protective effect on d-galactose (D-gal) induced skin aging in mice. The results showed that CPE had strong free radical scavenging, reducing and metal chelating activities. CPE can increase the activity of catalase (CAT), glutathione peroxidase (GSH-Px), peroxidase (POD), superoxide dismutase (SOD), and the content of glutathione (GSH), decrease the content of malondialdehyde (MDA). Moreover, CPE can decrease the gene and protein expression of matrix metalloproteinase 1a (MMP-1a) and matrix metalloproteinase 3 (MMP-3) and increase the expression of transforming growth factor-ß (TGF-ß) and tissue inhibitor of metalloproteinase 1 (TIMP-1) of mouse skin. Histopathological analysis showed CPE reduced the collagen damage caused by D-gal, increased collagen synthesis and reduced its degradation to delay skin aging.

The Effect of Mask Wearing on the Accuracy of Radiology Reports in an Academic Hospital Setting.

PURPOSE: In response to the pandemic, some public health agencies recommend the wearing of surgical masks in indoor spaces including radiology common reporting rooms. We aim to demonstrate whether mask wearing may lead to increased errors incidence in radiology reports. MATERIALS AND METHODS: Our prospective studywas conveyed in 2 parts. Firstly, the participants were surveyed if they believed that mask affected dictation. Then participants performed a dictation: they read artificial radiology reports using a commercial voice recognition (VR) system. They performed this task 5 times, each time donning a different mask in random order: a surgical mask, surgical visor, N-95, combination of 2 surgical masks and no mask. Error rates were compared with the Friedman test followed by pairwise Wilcoxon with bootstrapping. Multivariate Poisson regression was performed to test for interaction effects between potential predictors. RESULTS: 52 members of an academic radiology department participatedin the study (January - March 2021) . 65.4% of survey participants did not think or were not sure whether mask wearing could affect dictation process. Treating the no-mask condition as baseline, our study found that mean error rates significantly increased up to 2 times the baseline rate when a surgical mask, surgical visor, N-95 or a combination of 2 masks was donned (p < 0.0001). No significant differences in error rates were found between the different mask types (p > 0.05). Error rates were higher for participants with shorter VR training time (p < 0.0001) or who were non-native English speakers (p < 0.0001). There were no interaction effects between mask type, VR training time or English nativity, suggesting these variables to be independent predictors for error rate. Academic rank did not significantly affect the error rate. CONCLUSION: radiologists underestimate the influence of masks on dictation accuracy. mask wearing may lead to significant increase in dictational errors.

ASPECTS estimation using dual-energy CTA-derived virtual non-contrast in large vessel occlusion acute ischemic stroke: a dose reduction opportunity for patients undergoing repeat CT?

PURPOSE: Recent studies have shown the feasibility of dual-energy CT (DECT) virtual non-contrast (VNC) for determining infarct extent. In this study, patients presenting with large-vessel occlusion (LVO) acute ischemic stroke (AIS), we assess whether ASPECTS on DECTA-VNC differs from non-contrast CT (NCCT). METHODS: After IRB approval, LVO-AIS patients undergoing NCCT and DECTA between October 2016 and September 2018 were retrospectively reviewed. DECTA-VNC images were derived using Syngo.via (Siemens, Erlangen, Germany). ASPECTS was scored by two blinded neuroradiologists. Square-weighted kappa statistic, diagnostic performance, Wilcoxon signed-rank tests between groups, and CT doses were calculated. RESULTS: Fifty-one patients met inclusion criteria, with median age of 76 (IQR 67-82); 26/51 (51%) were female. Median time between last-known-well and CT was 120 min (IQR 60-252). DECTA-VNC ASPECTS score differed by ≤ 1 from consensus NCCT in 49/51 (96%) patients for reader 1 and in 46/51 (90%) for reader 2. ASPECTS on DECTA-SI and consensus NCCT differed by ≤ 1 in 45/51 (88%) for both readers. On a per ASPECTS-region basis, DECTA-VNC had 87% sensitivity, 95% specificity, 0.82% PPV, and 0.96% NPV. ASPECTS inter-rater agreement was highest for DECTA-VNC (κ = 0.71), DECTA-SI (κ = 0.48), and NCCT (κ = 0.40). NCCT median CTDIvol was 63.7 mGy (IQR 60.7-67.2); DLP was 1060.0 mGy·cm (IQR 981.0-1151.5). DECTA-VNC dose was lower: median CTDIvol was 20.9 mGy (IQR 19.8-22.2); DLP was 804.1 (IQR 691.6-869.4), p < 0.0001. CONCLUSION: DECTA-derived VNC yielded similar ASPECTS scores as NCCT and is therefore non-inferior in early ischemia-related low attenuation edema/infarct detection in acute LVO-AIS patients. Further evaluation of the role of DECTA-VNC in AIS imaging is warranted.

Comparative iTRAQ Proteomics Reveals Multiple Effects of Selenium Yeast on Dairy Cows in Parturition.

The effects of prepartum dietary supplementation with selenium yeast on low abundant plasma proteins in postpartum dairy cows are not known. In this study, 24 healthy parturient dairy cows were divided into two groups (group C, a control group, and group T, a selenium treatment group). Low abundance proteins were extracted from plasma samples of calving cows, and 542 proteins were identified by isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis. Dietary supplementation with selenium yeast caused differential abundance of 48 proteins with a fold change of more than 1.2 or less than 0.83 (p < 0.05); 14 proteins were upregulated and 34 were downregulated. The top five gene ontology (GO) enrichment terms for the differentially expressed proteins were protein homotetramerization (or tetramerization), defense response to bacteria or fungus, acute-phase reactions, nucleotide catabolic process, and positive regulation of lipid metabolic process. All proteins involved in acute-phase reactions were downregulated, indicating that selenium ameliorates systemic inflammation. The vast majority of proteins involved in the defense response to microorganisms were downregulated, thereby affecting innate immunity. The decreased abundance of apolipoprotein A-I and apolipoprotein C-II, critical proteins for positive regulation of lipid metabolism, indicated that selenium may optimize lipid metabolism. The iTRAQ results showed that prenatal supplementation with yeast selenium can relieve systemic inflammation after parturition. Moreover, selenium may reduce the effects of metabolic diseases, which can improve glyconeogenesis and prevent ketosis and fatty liver.

Characterization of mental health in cannabis dispensary users, using structured clinical interviews and standardized assessment instruments.

BACKGROUND: Cannabis is commonly used for its medical properties. In particular, cannabis is purported to have beneficial effects on a wide range of neuropsychiatric conditions. Studies assessing mental health in cannabis dispensary users typically evaluate symptoms using self-report check lists, which provide limited information about symptom severity, and whether subjects meet criteria for a psychiatric diagnosis. There is, therefore, a need for studies which assess mental health in dispensary users with standardized and well validated scientific instruments, such as those used in clinical drug trials. METHODS: One hundred medical cannabis users were recruited from a community dispensary. All subjects completed a structured clinical interview with the Mini-International Neuropsychiatric Interview (MINI). Subjects also completed the Perceived Stress Scale-10, PROMIS Fatigue Scale, PROMIS Sleep Disturbance Scale, Beck Depression Inventory, the Patient Health Questionnaire-15 and the Brief Pain Inventory. Details about cannabis use were also recorded. RESULTS: Lifetime prevalence of mental illness in this cohort was high, and a large proportion of subjects endorsed psychological symptoms. The proportion of subjects who met criteria for classification of a current psychiatric disorder was low for mood disorders, but high for anxiety disorders and substance abuse/dependence. Cannabis use differed between the main psychiatric conditions. CONCLUSIONS: The present results indicate that rates of mental illness may be high in medical cannabis dispensary users. Use of structured clinical assessments combined with standardized symptom severity questionnaires provide a feasible way to provide a more rigorous and detailed evaluation of conditions and symptoms in this population.

[Role of Sertoli cells in regulating spermatogenesis: An update].

Spermatogenesis is a complex process in which Sertoli cells play a pivotal role. Sertoli cells are closely related to testicular development and male fertility by providing nutrition for the proliferation of germ cells and maintaining the balance of microenvironment in seminiferous tubules. This article focuses on the roles of Sertoli cells in normal and abnormal spermatogenesis, outlines the association of spermatogenesis with the number and morphology of and protein expressions and secretions in Sertoli cells, and provides some reference for the studies of the gene therapy for male infertility and male contraception.

Genome-wide DNA methylation profile of prepubertal porcine testis.

The biological structure and function of the mammalian testis undergo important developmental changes during prepuberty and DNA methylation is dynamically regulated during testis development. In this study, we generated the first genome-wide DNA methylation profile of prepubertal porcine testis using methyl-DNA immunoprecipitation (MeDIP) combined with high-throughput sequencing (MeDIP-seq). Over 190 million high-quality reads were generated, containing 43642 CpG islands. There was an overall downtrend of methylation during development, which was clear in promoter regions but less so in gene-body regions. We also identified thousands of differentially methylated regions (DMRs) among the three prepubertal time points (1 month, T1; 2 months, T2; 3 months, T3), the majority of which showed decreasing methylation levels over time. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed that many genes in the DMRs were linked with cell proliferation and some important pathways in porcine testis development. Our data suggest that DNA methylation plays an important role in prepubertal development of porcine testis, with an obvious downtrend of methylation levels from T1 to T3. Overall, our study provides a foundation for future studies and gives new insights into mammalian testis development.

[Effects of retinol on expressions of epidermal growth factor, stem cell factor, colony-stimulating factor 1 and leukemia inhibitory factor in human umbilical cord-derived mesenchymal stem cells].

OBJECTIVE: To investigate effects of retinol on the expressions of epidermal growth factor (EGF), stem cell factor (SCF), colony-stimulating factor 1 (CSF1) and leukemia inhibitory factor (LIF) in cultured human umbilical-derived mesenchymal stem cells (UCMSCs). METHODS: Human UCMSCs were isolated from human umbilical cord and identified for immunophenotypes. The cells were then cultured in DMEM/F12 media supplemented with 12% fetal bovine serum (FBS), 12% FBS+1 µmol/L retinol, 15% knockout serum replacement (KSR) and 15% KSR+ 1 µmol/L retinol. The expressions of the cytokines EGF, SCF, CSF1 and LIF in the cells were detected using RT-PCR and ELISA. RESULTS: The isolated cells exhibited characteristic immunophenotypes of human UCMSCs and expressed EGF, CSF1 and SCF at both mRNA and protein levels but not LIF protein. Retinol (1 µmol/L) significantly promoted the expressions of SCF and CSF1 at both mRNA and protein levels but did not result in changes of EGF and LIF expressions in human UCMSCs. CONCLUSION: Retinol at the concentration of 1 µmol/L can promote expression of SCF and CSF1 in human UCMSCs in vitro.

CRISPR/Cas9 genome editing technique and its application in site-directed genome modification of animals.

CRISPR/Cas system, which uses CRISPR RNAs (crRNAs) to guide Cas nuclease to silence invading nucleic acids, is self-defense system against exogenous virus or plasmid in bacteria and archaea. Through molecular modification, the typeⅡCRISPR/Cas system has become a highly efficient site-directed genome editing technique, which is simpler than zinc-finger nucleases (ZFNs) and transcription activator like effector nucleases (TALENs) and easier to be designed and applied. In this review, we summarize the evolutionary history of CRISPR/Cas9 system, the working principle and modification process of type Ⅱ CRISPR/Cas and its application in animal genome modification. We also analyze the existing problems and improvement program of the CRISPR/Cas9 system as well as its application prospect combined with successful cases, which may provide innovative perspectives on improving animal traits and establishing animal models of human diseases.