Metabolic skeletal disorders associated with impaired bone formation are a major clinical challenge. One approach to treat these defects is to silence bone-formation-inhibitory genes by small interference RNAs (siRNAs) in osteogenic-lineage cells that occupy the niche surrounding the bone-formation surfaces. We developed a targeting system involving dioleoyl trimethylammonium propane (DOTAP)-based cationic liposomes attached to six repetitive sequences of aspartate, serine, serine ((AspSerSer)(6)) for delivering siRNAs specifically to bone-formation surfaces. Using this system, we encapsulated an osteogenic siRNA that targets casein kinase-2 interacting protein-1 (encoded by Plekho1, also known as Plekho1). In vivo systemic delivery of Plekho1 siRNA in rats using our system resulted in the selective enrichment of the siRNAs in osteogenic cells and the subsequent depletion of Plekho1. A bioimaging analysis further showed that this approach markedly promoted bone formation, enhanced the bone micro-architecture and increased the bone mass in both healthy and osteoporotic rats. These results indicate (AspSerSer)(6)-liposome as a promising targeted delivery system for RNA interference-based bone anabolic therapy.

The research of carbon-based antivirals is still in its infancy, and their development into safe and effective carbon dots (CDs) with antiviral activity at multiple points in the life cycle of the virus remains to be explored. Here, we report a one-step method to apply curcumin in order to prepare of uniform and stable cationic carbon dots (CCM-CDs) with antiviral properties. The inhibitory effect of CCM-CDs on viral replication was studied by using porcine epidemic diarrhea virus (PEDV) as a coronavirus model. PEDV is applied as a coronavirus model to study the antiviral effect of as-prepared CCM-CDs on its replication. The cationic CCM-CDs treatment is found obviously to inhibit the proliferation of PEDV compared with the common CDs (EDA-CDs). The CCM-CDs treatment can change the structure of surface protein in viruses, thereby inhibiting viral entry. It can also suppresses the synthesis of negative-strand RNA of the virus, the budding of the virus, and the accumulation of reactive oxygen species by PEDV. Furthermore, CCM-CDs treatment is also found to suppress viral replication by stimulating the production of interferon-stimulating genes (ISGs) and proinflammatory cytokines. These results offer theoretical support for the development of CCM-CDs as a hopeful antiviral drug for the treatment of coronavirus infections, including PEDV.

Development of novel antiviral reagents is of great importance for the control of virus spread. Here, AgS nanoclusters (NCs) were proved for the first time to possess highly efficient antiviral activity by using porcine epidemic diarrhea virus (PEDV) as a model of coronavirus. Analyses of virus titers showed that AgS NCs significantly suppressed the infection of PEDV by about 3 orders of magnitude at the noncytotoxic concentration at 12 h postinfection, which was further confirmed by the expression of viral proteins. Mechanism investigations indicated that AgS NCs treatment inhibits the synthesis of viral negative-strand RNA and viral budding. AgS NCs treatment was also found to positively regulate the generation of IFN-stimulating genes (ISGs) and the expression of proinflammation cytokines, which might prevent PEDV infection. This study suggest the novel underlying of AgS NCs as a promising therapeutic drug for coronavirus.

No abstract

Developing nanomaterials-based antimicrobial agents has shown a widespread promise. In this study, silver nanoparticle-modified graphene oxide (GO-AgNPs) nanocomposites were self-assembled via interfacial electrostatic force. By using the porcine reproductive and respiratory syndrome virus (PRRSV) as a pattern, the antiviral effect of the as-prepared GO-AgNPs nanocomposites on the replication of virus was investigated. The results indicated that exposure with GO-AgNPs nanocomposites could obviously suppress PRRSV infection. It was found that GO-AgNPs nanocomposites exhibited a better inhibitory effect compared with AgNPs and GO. By selecting the porcine epidemic diarrhea virus (PEDV) as a contrast virus, GO-AgNPs nanocomposites were proven to have a broad antiviral activity. Mechanism studies showed that GO-AgNPs nanocomposites might prevent PRRSV from entering the host cells, with 59.2% inhibition efficiency. Meanwhile, GO-AgNPs nanocomposite treatment enhances the production of interferon-α (IFN-α) and IFN-stimulating genes (ISGs), which can directly inhibit the proliferation of virus. Taken together, this study reports a new type of antiviral agent and provides a promising pharmaceutical agent for treating infection by the highly pathogenic PRRSV. Moreover, it may provide novel ideas for the research and development of antiviral formulations based on nanocomposites and extend their applications in biological systems.

No abstract

Porcine deltacoronavirus (PDCoV) is an emerging swine coronavirus that causes diarrhea in piglets. Since the first outbreak of PDCoV in the United States in 2014, this novel porcine coronavirus has been detected in South Korea, Canada, Mexico, Thailand, and China. In this study, a Chinese PDCoV strain, designated CHN-HN-2014, was isolated from piglets with severe diarrhea on a pig farm in Henan Province, China, and examined with a specific immunofluorescence assay and electron microscopy. Genomic analysis showed that CHN-HN-2014 shares 91.6%-99.4% nucleotide identity with other known PDCoV strains. The pathogenicity of CHN-HN-2014 was further investigated in 5-day-old and 21-day-old piglets. Both kinds of piglets developed clear clinical symptoms, including vomiting, anorexia, lethargy, and severe diarrhea, by 2days postinoculation (DPI), and diarrhea persisted for about 5-6 days. Viral shedding was detected in rectal swabs until 14 DPI in challenged 5-day-old pigs and until 18 DPI in challenged 21-day-old pigs. At necropsy at 4 DPI, macroscopic and microscopic lesions were observed and viral antigen was detected in the small intestines with immunohistochemical staining. These data demonstrate that Chinese PDCoV strain CHN-HN-2014 shares high nucleotide identity with previously reported PDCoV strains and is pathogenic in 5-day-old and 21-day-old piglets.

The hepatitis B virus X protein (HBx) plays an important role in the development of hepatocellular carcinoma (HCC). The relationship was examined between HBV antigens and IAP (inhibitor of apoptosis) family in development of HCC. The expression levels of HBV antigens (HBsAg, HBcAg, and HBxAg) and members of the IAP family (survivin, XIAP, cIAP-1, and cIAP-2) were detected immunohistochemically in tissues from 34 cases of HCC and 30 cases of liver cirrhosis. The positive rate of survivin was higher than these three molecules in all three tissue types (P < 0.05). The positive rates of HBxAg and survivin were high in HCC (76.5% and 88.2%), paratumor (85.3% and 91.2%), and liver cirrhosis (100% and 93.3%) tissues, with no significant differences between the survivin- and HBxAg-positive rates (each P > 0.05). To examine the effect of HBx on survivin expression, plasmid pCMV-X (encoding the HBx gene) was transfected transiently with or without plasmid pcDNA3-sur (encoding the survivin gene) into H7402 hepatoma cells and L-O2 human normal liver cells. Cells over-expressing HBx alone showed increased apoptosis along with a dose-dependent increase in survivin levels. However, co-expression of survivin inhibited the HBx-induced apoptosis. To examine the effect of HBx on survivin in hepatoma cells without apoptosis, plasmid pCMV-X was transfected stably into human hepatoma H7402 cells and L-O2 cells. These H7402-X and L-O2-X cells showed high-level expression of both HBx and survivin, but did not show apoptosis. The addition of pSilencer 3.0-X, an RNAi vector targeting the HBx gene, reduced the expression levels of survivin protein in H7402-X cells. Collectively, these data demonstrate that HBx upregulates survivin expression in hepatoma tissues, suggesting that HBx and survivin may both be involved in carcinogenesis of HCC.