PSB0739 inhibits formation of semen-derived amyloid fibril / 南方医科大学学报

OBJECTIVE@#To explore the inhibitory effect of PSB0739 on the formation of semen-derived amyloid fibrils.@*METHODS@#PAP248-286 (440 μmol/L) was incubated with PSB0739 at different concentrations, and at different time points of incubation, aliquots were taken from each sample for Congo red staining to detect the formation of amyloid fibers. The morphology of amyloid fibrils incubated in the presence or absence of PSB0739 was visualized using transmission electron microscope. The effect of PSB0739 on amyloid fibril formation was determined using virus infection assays at different time points, and the surface charges of amyloid fibril incubated with PSB0739 were calculated using a Zeta potentiometer. The cytotoxicity of PSB0739 in Hela cells was determined using MTT assay. The antiviral effect of PSB0738 against HIV- 1 was evaluated by infection assay.@*RESULTS@#PSB0739 inhibited SEVI fibril formation in a dose-dependent manner . At 48 h of incubation, 220 μmol/L of PSB0739 obviously inhibited the formation of amyloid fibrils in 440 μmol/L of SEVI. Transmission electron microscopy revealed that 220 μmol/L PSB0739 prevented PAP248- 286 (440 μmol/L) from forming amyloid fibrils. PSB0739 antagonized SEVI-mediated enhancement of HIV-1 infection, and 1760 μmol/L of PSB0739 completely reversed the positive charge of SEVI ( < 0.05). PSB0739 below the concentration of 62.5 μmol/L showed no obvious cytotoxicity in Hela cells (>0.05). PSB0739 showed a direct anti-HIV activity with an IC of 21.77±5.15 μmol/L.@*CONCLUSIONS@#PSB0739 can inhibit the formation of semen-derived amyloid fibrils .

The cloning of mouse nicotinamide mononucleotide adenylyl-transferase gene and the detecting of its expression / 中华老年医学杂志

Objective To construct eukaryotic expressing vector of the mouse NMNAT1 (nicotinamide mononucleotide adenylyl-transferase) gene and examine its ability to express the NMNAT1 gene in Hela cells.Methods The full-length NMNAT1 cDNA sequence was amplified by PCR and cloned into the plasmid of T-vector and then to pcDNA3.1 construct.The recombinant plasmid pcDNA3.1-NMNAT1 was identified by DNA sequencing and then transfected with Lipofectamine2000 into Hela cells.The expression of NMNAT1 was detected by real time quantitative PCR (qPCR) and Western blot after 48 h transfection.Results The recombinant eukaryotic vector carrying NMNAT1 gene was constructed successfully in a match with database and this vector could up-regulate the expression of the NMNAT1 gene both in mRNA and protein levels in Hela cells.Conclusions The eukaryotic vector carrying NMNAT1 gene (pcDNA3.1-NMNAT1) enhances the expression of NMNAT1 gene.