Interferon-Inducible Guanylate-Binding Protein 5 Inhibits Replication of Multiple Viruses by Binding to the Oligosaccharyltransferase Complex and Inhibiting Glycoprotein Maturation.

Viral infection induces production of type I interferons and expression of interferon-stimulated genes (ISGs) that play key roles in inhibiting viral infection. Here, we show that the ISG guanylate-binding protein 5 (GBP5) inhibits N-linked glycosylation of key proteins in multiple viruses, including SARS-CoV-2 spike protein. GBP5 binds to accessory subunits of the host oligosaccharyltransferase (OST) complex and blocks its interaction with the spike protein, which results in misfolding and retention of spike protein in the endoplasmic reticulum likely due to decreased N -glycan transfer, and reduces the assembly and release of infectious virions. Consistent with these observations, pharmacological inhibition of the OST complex with NGI-1 potently inhibits glycosylation of other viral proteins, including MERS-CoV spike protein, HIV-1 gp160, and IAV hemagglutinin, and prevents the production of infectious virions. Our results identify a novel strategy by which ISGs restrict virus infection and provide a rationale for targeting glycosylation as a broad antiviral therapeutic strategy. Highlights: The interferon-stimulated gene GBP5 is induced by SARS-CoV-2 infection in vitro and in vivo.ER-localized GBP5 restricts N-linked glycosylation of SARS-CoV-2 spike protein, leading to protein misfolding and preventing transport to the Golgi apparatus.GBP5 binds to OST complex accessory proteins and potentially blocks access of the catalytic subunit to the spike protein.GBP5 inhibits N-glycosylation of key proteins in multiple viruses, including SARS-CoV-2Pharmacological inhibition of OST blocks host cell infection by SARS-CoV-2, variants of concern, HIV-1, and IAV. Significance: Viral infection induces production of type I interferons and expression of interferon-stimulated genes (ISGs) that play key roles in inhibiting viral infection. We found that the interferon-stimulated gene GBP5 is induced by SARS-CoV-2 infection in vitro and in vivo. GBP5 inhibits N-glycosylation of key proteins in multiple viruses, including SARS-CoV-2. Importantly, pharmacological inhibition of Oligosaccharyltransferase (OST) Complex blocks host cell infection by SARS-CoV-2, variants of concern, HIV-1, and IAV, indicating future translational application of our findings.

Urushiol modified epoxy acrylate as UV spray painting oriental lacquer ink.

As a natural "water-based" polymer composite material, oriental lacquer is often referred to as the "king of coatings" and is used as a coating in the defense industry, chemical industry, petroleum industry, metallurgy and mining industry, textile painting and dyeing industry, pharmaceutical industry, as well as the protection of ancient buildings and cultural relics. However, the development of modern industrialization is greatly hindered by the high viscosity of oriental lacquer, the difficulty of spraying, the long drying cycle, and the seriousness of allergenicity. Herein, based on the principle of oriental lacquer and the characteristics of prepolymer in ink, we developed a new prepolymer for modulating UV oriental lacquer ink and explored the feasibility of using it as a raw material for UV spray painting. In this study, lacquer phenol was extracted from oriental lacquer and modified with epoxy acrylate by a simple mechanical compounding method to obtain lacquer epoxy acrylate. Moreover, the UV spray painting oriental lacquer ink was also prepared by using it as the main film-forming substance. The orthogonal experiment method was used to optimize the best formulation of UV spray painting oriental lacquer ink by using nozzle passability, viscosity and curing time as test indexes. Meanwhile, the film properties of UV spray painting oriental lacquer inks were also evaluated. The test results show that the UV spray painting oriental lacquer ink prepared with urushiol epoxy acrylate has better dispersion, excellent paint film performance, and solves the problem that oriental lacquer cannot be printed. This present work shows that urushiol epoxy acrylate as a new type of prepolymer has broad application prospects in the actual preparation of UV inks.

PCIF1-mediated deposition of 5'-cap N6,2'-O-dimethyladenosine in ACE2 and TMPRSS2 mRNA regulates susceptibility to SARS-CoV-2 infection.

Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to be a major health problem worldwide. Due to the fast emergence of SARS-CoV-2 variants, understanding the molecular mechanisms of viral pathogenesis and developing novel inhibitors are essential and urgent. Here, we investigated the potential roles of N6,2'-O-dimethyladenosine (m6Am), one of the most abundant modifications of eukaryotic messenger ribonucleic acid (mRNAs), in SARS-CoV-2 infection of human cells. Using genome-wide m6Am-exo-seq, RNA sequencing analysis, and Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 genome editing, we demonstrate that phosphorylated C-terminal domain (CTD)-interacting factor 1 (PCIF1), a cap-specific adenine N6-methyltransferase, plays a major role in facilitating infection of primary human lung epithelial cells and cell lines by SARS-CoV-2, variants of concern, and other coronaviruses. We show that PCIF1 promotes infection by sustaining expression of the coronavirus receptors angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) via m6Am-dependent mRNA stabilization. In PCIF1-depleted cells, both ACE2/TMPRSS2 expression and viral infection are rescued by re-expression of wild-type, but not catalytically inactive, PCIF1. These findings suggest a role for PCIF1 and cap m6Am in regulating SARS-CoV-2 susceptibility and identify a potential therapeutic target for prevention of infection.

Role of PCIF1-mediated 5'-cap N6-methyladeonsine mRNA methylation in colorectal cancer and anti-PD-1 immunotherapy.

Adenosine N6-methylation (m6A) and N6,2'-O-dimethylation (m6Am) are regulatory modifications of eukaryotic mRNAs. m6Am formation is catalyzed by the methyl transferase phosphorylated CTD-interacting factor 1 (PCIF1); however, the pathophysiological functions of this RNA modification and PCIF1 in cancers are unclear. Here, we show that PCIF1 expression is upregulated in colorectal cancer (CRC) and negatively correlates with patient survival. CRISPR/Cas9-mediated depletion of PCIF1 in human CRC cells leads to loss of cell migration, invasion, and colony formation in vitro and loss of tumor growth in athymic mice. Pcif1 knockout in murine CRC cells inhibits tumor growth in immunocompetent mice and enhances the effects of anti-PD-1 antibody treatment by decreasing intratumoral TGF-ß levels and increasing intratumoral IFN-γ, TNF-α levels, and tumor-infiltrating natural killer cells. We further show that PCIF1 modulates CRC growth and response to anti-PD-1 in a context-dependent mechanism with PCIF1 directly targeting FOS, IFITM3, and STAT1 via m6Am modifications. PCIF1 stabilizes FOS mRNA, which in turn leads to FOS-dependent TGF-ß regulation and tumor growth. While during immunotherapy, Pcif1-Fos-TGF-ß, as well as Pcif1-Stat1/Ifitm3-IFN-γ axes, contributes to the resistance of anti-PD-1 therapy. Collectively, our findings reveal a role of PCIF1 in promoting CRC tumorigenesis and resistance to anti-PD-1 therapy, supporting that the combination of PCIF1 inhibition with anti-PD-1 treatment is a potential therapeutic strategy to enhance CRC response to immunotherapy. Finally, we developed a lipid nanoparticles (LNPs) and chemically modified small interfering RNAs (CMsiRNAs)-based strategy to silence PCIF1 in vivo and found that this treatment significantly reduced tumor growth in mice. Our results therefore provide a proof-of-concept for tumor growth suppression using LNP-CMsiRNA to silence target genes in cancer.

[Using ultracision-harmonic scalpel as major instrument for tonsillectomy via nasal endoscope (a report of 31 cases)].

OBJECTIVE: To investigate the feasibility of applying ultracision-harmonic scalpel in tonsillectomy. METHOD: Thirty-one cases treated with tonsillectomy which applied with nasal endoscope and ultracision-harmonic scalpel (ultrasonic scalpel group) and 31 cases treated with routine operation (routine group) respectively. The operating time and blood loss during operation and the degree of pain and the region of inflammation humidity as well as hospitalization days were observed. RESULT: All indexes in ultrasonic scalpel group were lower than that in the routine group, including operating time and blood loss during operation, and they were obviously difference in statistics (P < 0.01). CONCLUSION: The application of ultracision-harmonic scalpel in tonsillectomy was safe, efficient and feasible, and would have a nice application prospect during ear-nose-throat operation.