Transcriptome Analysis of the Midgut of the Chinese Oak Silkworm Antheraea pernyi Infected with Antheraea pernyi Nucleopolyhedrovirus.

The Antheraea pernyi nucleopolyhedrovirus (ApNPV) is an exclusive pathogen of A. pernyi. The intense interactions between ApNPV and A. pernyi cause a series of physiological and pathological changes to A. pernyi. However, no detailed report exists regarding the molecular mechanisms underlying the interactions between ApNPV and A. pernyi. In this study, four cDNA libraries of the A. pernyi midgut, including two ApNPV-infected groups and two control groups, were constructed for transcriptomic analysis to provide new clues regarding the molecular mechanisms that underlie these interactions. The transcriptome of the A. pernyi midgut was de novo assembled using the Trinity platform because of the lack of a genome resource for A. pernyi. Compared with the controls, a total of 5,172 differentially expressed genes (DEGs) were identified, including 2,183 up-regulated and 2,989 down-regulated candidates, of which 2,965 and 911 DEGs were classified into different GO categories and KEGG pathways, respectively. The DEGs involved in A. pernyi innate immunity were classified into several categories, including heat-shock proteins, apoptosis-related proteins, serpins, serine proteases and cytochrome P450s. Our results suggested that these genes were related to the immune response of the A. pernyi midgut to ApNPV infection via their essential roles in regulating a variety of physiological processes. Our results may serve as a basis for future research not only on the molecular mechanisms of ApNPV invasion but also on the anti-ApNPV mechanism of A. pernyi.

iPSCs are transcriptionally and post-transcriptionally indistinguishable from fESCs.

Induced pluripotent stem cells (iPSCs) are generated by reprogramming mouse or human somatic cells to a pluripotent state by introducing key transcription factors and have great therapeutic potential. It has been illustrated that the transcriptional and post-transcriptional profiles of nuclear-transferred embryonic stem cells (ntESCs) is identical to those of embryonic stem cells derived from fertilized blastocysts (fESCs). Although iPSCs seem to be indistinguishable from fESCs, the degree of transcriptomic and proteomic similarity among iPSCs, ntESCs, and fESCs has not yet been elucidated completely. To investigate whether iPSCs and fESCs have similar therapeutic potential, we compared mRNA and protein pro?les of mouse iPSC, ntESCs, and matching fESCs lines using microarray technology, iTRAQ method, and bioinformatic analyses. Real-time PCR, two-dimensional LC, and MS/MS analyses were further conducted to study the expression of speci?c transcripts and identify and quantitate 929 proteins. Our results demonstrate that, like ntESCs, the iPSC and matching fESCs lines have very similar transcriptional and protein expression profiles. This is consistent with their similar developmental potential.

A novel polymer-lipid hybrid nanoparticle for efficient nonviral gene delivery.

AIM: To develop a novel non-viral vector with high transfection efficiency and low cytotoxicity. METHODS: Poly (ethylene glycol)-distearoylphosphatidylethanolamine (PEG-DSPE) was incorporated into polymer-lipid hybrid nanoparticles (PLN) to construct a PEG-DSPE modified long circulating PLN (L-PLN). The L-PLN was prepared by the emulsifying-solvent evaporation method, L-PLN and L-PLN/DNA complexes were characterized. Both HEK293 and MDA-MB-231 cells transfected by L-PLN/DNA complexes were observed under a fluorescence microscope. The transfection efficiency of the complexes to HEK293 cells was further evaluated by flow cytometry. RESULTS: The GFP fluorescence intensity in HEK293 cells transfected by the L-PLN/DNA complexes (N/P=10) was about 37.2%, which was higher than those transfected by PLN alone or commercial Lipofectamine 2000. The L-PLN exhibited minimal toxicity at a low N/P ratio compared with other vectors. CONCLUSION: L-PLN as a novel gene delivery system, has higher transfection efficiency and acceptable cytotoxicity compared to the corresponding PLN, which is beneficial for the development of non-viral gene transfer vectors and may offer an alternative strategy for the future gene therapy.