TRIM6 facilitates SARS-CoV-2 proliferation by catalyzing the K29-typed ubiquitination of NP to enhance the ability to bind viral genomes.

The Nucleocapsid Protein (NP) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is not only the core structural protein required for viral packaging, but also participates in the regulation of viral replication, and its post-translational modifications such as phosphorylation have been shown to be an important strategy for regulating virus proliferation. Our previous work identified NP could be ubiquitinated, as confirmed by two independent studies. But the function of NP ubiquitination is currently unknown. In this study, we first pinpointed TRIM6 as the E3 ubiquitin ligase responsible for NP ubiquitination, binding to NP's CTD via its RING and B-box-CCD domains. TRIM6 promotes the K29-typed polyubiquitination of NP at K102, K347, and K361 residues, increasing its binding to viral genomic RNA. Consistently, functional experiments such as the use of the reverse genetic tool trVLP model and gene knockout of TRIM6 further confirmed that blocking the ubiquitination of NP by TRIM6 significantly inhibited the proliferation of SARS-CoV-2. Notably, the NP of coronavirus is relatively conserved, and the NP of SARS-CoV can also be ubiquitinated by TRIM6, indicating that NP could be a broad-spectrum anti-coronavirus target. These findings shed light on the intricate interaction between SARS-CoV-2 and the host, potentially opening new opportunities for COVID-19 therapeutic development.

The PLAG1 mRNA expression analysis among genetic variants and relevance to growth traits in Chinese cattle.

Pleomorphic adenoma gene 1 (PLAG1) encodes a developmentally regulated zinc finger protein, locating in growth-related QTNs. The mRNA expression of this gene was investigated in different tissues and from two different developmental periods, whilst to explore the functions of PLAG1 in growth traits of cattle. The results showed that PLAG1 was expressed in all examined tissues. However, PLAG1 expression levels in all examined tissues were significantly different between the 5-month fetus and 36-month adult cattle. Our juvenile results indicated PLAG1 is primarily expressed in embryonic tissues of Chinese cattle. Furthermore, two variations were identified. Association analysis revealed that the two variations were associated with growth traits (p < 0.05 or p < 0.01). These new findings provide a comprehensive overview of the critical roles of PLAG1 in growth traits modulation and can be highlighted as candidate molecular markers in cattle breeding.

Morus nigra L. leaves improve the meat quality in finishing pigs.

This study was conducted to evaluate effects of dietary Mulberry leaves on growth performance, carcass traits and meat quality in finishing pigs. Here, a total of 72 crossbred [(Landrace × Yorkshire) × Duroc] pigs with an average initial body weight of 70.03 ± 0.48 kg were used in this 45-day feeding trial. The pigs were randomly divided into three groups (6 pigs/pen and 4 replicates/group). Dietary treatments included a control diet (without any Mulberry leaves) and diets supplemented with 5% non- or fermented Mulberry leaf powder (MF or FMF respectively). The present findings indicated that compared with the control group, administration of MF or FMF significantly improved gain: feed ratio (p < .05) and reduced the backfat thickness (p < .05). Meanwhile, dietary MF and FMF significantly enhanced triglyceride deposition in Longissimus dorsi muscles (p < .05). Besides, both of MF and FMF could effectively improve the antioxidant capacity by increasing the content of T-AOC and SOD in serum and reduce the rancidity of pork. In conclusion, supplementary MF and FMF could promote gain: feed ratio, reduce backfat thickness, increase fat deposition in muscle and reduce the rancidity of pork.

Ciclopirox inhibits SARS-CoV-2 replication by promoting the degradation of the nucleocapsid protein.

The nucleocapsid protein (NP) plays a crucial role in SARS-CoV-2 replication and is the most abundant structural protein with a long half-life. Despite its vital role in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) assembly and host inflammatory response, it remains an unexplored target for drug development. In this study, we identified a small-molecule compound (ciclopirox) that promotes NP degradation using an FDA-approved library and a drug-screening cell model. Ciclopirox significantly inhibited SARS-CoV-2 replication both in vitro and in vivo by inducing NP degradation. Ciclopirox induced abnormal NP aggregation through indirect interaction, leading to the formation of condensates with higher viscosity and lower mobility. These condensates were subsequently degraded via the autophagy-lysosomal pathway, ultimately resulting in a shortened NP half-life and reduced NP expression. Our results suggest that NP is a potential drug target, and that ciclopirox holds substantial promise for further development to combat SARS-CoV-2 replication.

The E3 ligase TRIM22 restricts SARS-CoV-2 replication by promoting proteasomal degradation of NSP8.

Replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome is mediated by a complex of non-structural proteins (NSPs), of which NSP7 and NSP8 serve as subunits and play a key role in promoting the activity of RNA-dependent RNA polymerase (RdRp) of NSP12. However, the stability of subunits of the RdRp complex has rarely been reported. Here, we found that NSP8 was degraded by the proteasome in host cells, and identified tripartite motif containing 22 (TRIM22) as its E3 ligase. The interferon (IFN) signaling pathway was activated upon viral invasion into host cells, and TRIM22 expression increased. TRIM22 interacted with NSP8 and ubiquitinated it at Lys97 via K48-type ubiquitination. TRIM22 overexpression significantly reduced viral RNA and protein levels. Knockdown of TRIM22 enhanced viral replication. This study provides a new explanation for treating patients suffering from SARS-CoV-2 with IFNs and new possibilities for drug development targeting the interaction between NSP8 and TRIM22.IMPORTANCENon-structural proteins (NSPs) play a crucial role in the replication of severe acute respiratory syndrome coronavirus 2, facilitating virus amplification and propagation. In this study, we conducted a comprehensive investigation into the stability of all subunits comprising the RNA-dependent RNA polymerase complex. Notably, our results reveal for the first time that NSP8 is a relatively unstable protein, which is found to be readily recognized and degraded by the proteasome. This degradation process is mediated by the host E3 ligase tripartite motif containing 22 (TRIM22), which is also a member of the interferon stimulated gene (ISG) family. Our study elucidates a novel mechanism of antiviral effect of TRIM22, which utilizes its own E3 ubiquitin ligase activity to hinder viral replication by inducing ubiquitination and subsequent degradation of NSP8. These findings provide new ideas for the development of novel therapeutic strategies. In addition, the conserved property of NSP8 raises the possibility of developing broad antiviral drugs targeting the TRIM22-NSP8 interaction.

New rural pension scheme, intergenerational interaction and rural family human capital investments.

Introduction: The new rural pension scheme (NRPS) can improve the quality of life for rural older adult individuals; however, can it have a spillover effect on rural household human capital investments through intergenerational interactions? Methods: Based on data from the China Family Panel Studies (CFPS) in 2010, 2012, 2014, 2016, and 2018 and from the perspective of intergenerational interactions, the spillover effect and influencing mechanism of the new rural insurance policy on rural household human capital investments are empirically tested. Results: The results show that the participation of families in the new rural insurance policy can significantly promote the human capital investments of rural families, and they are robust. Moreover, the spillover effect of this new policy is significantly different due to the gender, insurance phase, and family income of the insured. Through intergenerational interactions, the new rural insurance policy has an impact on the human capital investments of rural families from the material level of intergenerational economic support, housework and childcare for children and the nonmaterial level of old-age care cognition. Discussion: Therefore, continuing to promote the coverage of the new rural insurance policy and scientifically improving rural social security through publicity and education to promote benign intergenerational family interactions can improve the accumulation of human capital in rural areas.

Spike-mediated ACE2 down-regulation was involved in the pathogenesis of SARS-CoV-2 infection.

The ongoing global pandemic of Coronavirus disease 2019 (COVID-19) poses a serious threat to human health, with patients reportedly suffering from thrombus, vascular injury and coagulation in addition to acute and diffuse lung injury and respiratory diseases. Angiotensin converting enzyme 2 (ACE2) as the receptor for SARS-CoV-2 entry, is also an important regulator of renin-angiotensin system (RAS) homeostasis, which plays an unsettled role in the pathogenesis of COVID-19. Here, we demonstrated that SARS-CoV-2 Spike protein activated intracellular signals to degrade ACE2 mRNA. The decrease of ACE2 and higher level of angiotensin (Ang) II were verified in COVID-19 patients. High dose of Ang II induced pulmonary artery endothelial cell death in vitro, which was also observed in the lung of COVID-19 patients. Our finding indicates that the downregulation of ACE2 potentially links COVID-19 to the imbalance of RAS.

Multiomics approach reveals the ubiquitination-specific processes hijacked by SARS-CoV-2.

The Coronavirus Disease 2019 (COVID-19) caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a global pandemic that seriously threatens health and socioeconomic development, but the existed antiviral drugs and vaccines still cannot yet halt the spread of the epidemic. Therefore, a comprehensive and profound understanding of the pathogenesis of SARS-CoV-2 is urgently needed to explore effective therapeutic targets. Here, we conducted a multiomics study of SARS-CoV-2-infected lung epithelial cells, including transcriptomic, proteomic, and ubiquitinomic. Multiomics analysis showed that SARS-CoV-2-infected lung epithelial cells activated strong innate immune response, including interferon and inflammatory responses. Ubiquitinomic further reveals the underlying mechanism of SARS-CoV-2 disrupting the host innate immune response. In addition, SARS-CoV-2 proteins were found to be ubiquitinated during infection despite the fact that SARS-CoV-2 itself didn't code any E3 ligase, and that ubiquitination at three sites on the Spike protein could significantly enhance viral infection. Further screening of the E3 ubiquitin ligases and deubiquitinating enzymes (DUBs) library revealed four E3 ligases influencing SARS-CoV-2 infection, thus providing several new antiviral targets. This multiomics combined with high-throughput screening study reveals that SARS-CoV-2 not only modulates innate immunity, but also promotes viral infection, by hijacking ubiquitination-specific processes, highlighting potential antiviral and anti-inflammation targets.

Dietary supplementation of Morus nigra L. leaves decrease fat mass partially through elevating leptin-stimulated lipolysis in pig model.

ETHNOPHARMACOLOGICAL RELEVANCE: Mulberry leaves are the dry leaves of Morus nigra L. trees, which are widely cultivated in central and southern China. Mulberry has a long history of medicinal use, such as anti-stress, lowering blood glucose and anti-obesity. AIM OF THE STUDY: Explore the effects of mulberry leaves on fat deposition as well as the underlying mechanisms. MATERIALS AND METHODS: Total of 48 fattening pigs weighing about 70 kg were randomly allotted to normal diet or die supplemented with 5% (w/w) mulberry leave powder. Changes of fat mass, indicated by backfat thickness was measured with Piggyback tester, blood triglyceride and cholesterol were tested using commercial biochemical kits, serum hormones were estimated by ELISA, and leptin-related signaling activity were assessed using western-blot. RESULTS: Supplementation with Mulberry leaf feed (MF) significantly reduced serum triglyceride and free cholesterol concentrations and increased the ratio of high-density lipoprotein cholesterol (HDL-c) to low-density lipoprotein cholesterol (LDL-c), while serum glucose and free fatty acids remained unchanged. Dietary MF resulted in a significant reduction in the size of adipocytes and backfat thickness (P < 0.05). Accordingly, hormone-sensitive lipase (HSL) in backfat was significantly up-regulated and fatty acid synthase (FAS) was down-regulated by MF supplementation (both P < 0.05). Furthermore, MF supplementation significantly elevated circulating leptin and adiponectin without influencing serum insulin and glucocorticoid. Moreover, significantly higher leptin receptor (Leptin-R) and phosphorylated signal transducer and activator of transcription 3 (p-STAT3) were detected in MF-supplemented pigs, suggesting an enhanced leptin signaling induced by MF in the subcutaneous fat. CONCLUSIONS: Mulberry leaves have obvious anti-obesity effects, providing a theoretical basis for the development of mulberry leaves as a drug against obesity.

Effect of Fermented Corn-Soybean Meal on Serum Immunity, the Expression of Genes Related to Gut Immunity, Gut Microbiota, and Bacterial Metabolites in Grower-Finisher Pigs.

Fermented corn-soybean meal (fermented feed, FF) is commonly used in swine production, but the effects of FF on gut health remain unclear. In this study, serum immunity, mRNA abundances of antimicrobial peptides (AMPs) and Toll-like receptors (TLR1-9), bacterial abundance in the duodenum and colon, and colonic metabolic phenotypes were determined in crossbred barrows (Duroc × Landrace × Large White) fed FF or normal feed (unfermented feed, UF) (n = 6). When compared to the UF group, the results showed that serum levels of IgG and IgM were significantly increased in FF group pigs (P < 0.05). FF significantly decreased the abundances of Bacteroides and Verrucomicrobia in the duodenum and decreased the abundances of Bacteroides, Proteobacteria, and Verrucomicrobia in the colon while it significantly increased the abundances of Firmicutes and Actinobacteria (P < 0.05). Furthermore, a Spearman's correlation analysis showed that serum immunity and the expression of genes related to gut immunity were associated with bacterial strains at the family level. Moreover, differentially abundant colonic microbiota were associated with colonic metabolites. LC-MS data analyses identified a total of 1,351 metabolites that markedly differed between the UF and FF groups. C5-Branched dibasic acid metabolism was significantly upregulated whereas the purine metabolism was significantly downregulated (P < 0.05) in the colonic digesta of pigs in the FF meal group compared to the UF meal group. Collectively, these results indicated that FF meal could influence serum immunity and the expression of genes related to gut immunity, correlating with the gut microbiota and bacterial metabolites in grower-finisher pigs. This study may provide an alternative strategy for improving the intestinal health of grower-finisher pigs.

Fermented corn-soybean meal elevated IGF1 levels in grower-finisher pigs.

Fermentation has attracted increasing attention in pig industry, because of low costs and numerous benefits on pig growth and health as well as environmental improvement, although the mechanisms remain largely unknown. In the present study, fermented corn-soybean meal significantly improved average daily gain and gain:food ratio (P < 0.05). Fermented feed (FF) significantly increased insulin-like growth factor 1 (IGF1) transcription in liver (P < 0.05). Meanwhile, fermented meal significantly enhanced the binding of CCAAT/enhancer-binding protein beta (C/EBPß) to IGF1 promoter and C/EBPß expression in liver (both P < 0.05). FF tended to increase IGF1 proteins in liver and serum too (both 0.05 < P < 0.10). Meanwhile, FF slightly but significantly increased hepatic and circulating triglyceride and total cholesterol levels, as well as serum ratio of high-density to low-density cholesterol (all P < 0.05). Our data indicated that FF could significantly augment the binding of C/EBPß to IGF1 promoter and promote hepatic IGF1 expression and production, thus boost pig growth.