MiR-34a targets SIRT1 to reduce p53 deacetylation and promote sevoflurane inhalation anesthesia-induced neuronal autophagy and apoptosis in neonatal mice.

This study is to investigate the function of miR-34a and interactions between miR-34a, SIRT1, and p53 in sevoflurane-induced neuronal apoptosis and autophagy in neonatal mice. A mouse model was established by inhalation anesthesia with sevoflurane and injected with genetic reagents, followed by tests of learning and memory abilities and histological staining of the hippocampus. CCK-8 and AnnexinV/PI staining respectively measured the survival and apoptosis rates of primary hippocampal neurons cultured with sevoflurane. The expression levels of miR-34a, SIRT1, p53, Ac-p53, and autophagy- or apoptosis-related proteins were measured. Sevoflurane impaired the learning and memory abilities of mice, increased TUNEL-positive cells in their hippocampus, and hindered the survival of hippocampal neurons. Sevoflurane increased miR-34a, Bax, cleaved caspase-3, and the ratio of LC3-II/LC3-I and reduced SIRT1 and p62. MiR-34a overexpression promoted sevoflurane-induced neural damage, whereas SIRT1 inhibition or p53 upregulation counteracted the neuroprotection of miR-34a knockdown. SIRT1 was a target of miR-34a and promoted p53 deacetylation. MiR-34a promotes sevoflurane-stimulated neuronal apoptosis and autophagy in neonatal mice by inhibiting SIRT1 expression and subsequent p53 deacetylation.

Single-Frequency Precise Point Positioning Using Regional Dual-Frequency Observations.

High-precision and low-cost single-frequency precise point positioning (SF-PPP) has been attracting more and more attention in numerous global navigation satellite system (GNSS) applications. To provide the precise ionosphere delay and improve the positioning accuracy of the SF-PPP, the dual-frequency receiver, which receives dual-frequency observations, is used. Based on the serviced precise ionosphere delay, which is generated from the dual-frequency observations, the high-precision SF-PPP is realized. To further improve the accuracy of the SF-PPP and shorten its convergence time, the double-differenced (DD) ambiguity resolutions, which are generated from the DD algorithm, are introduced. This method avoids the estimation of fractional cycle bias (FCB) for the SF-PPP ambiguity. Here, we collected data from six stations of Shanghai China which was processed, and the corresponding results were analyzed. The results of the dual-frequency observations enhanced SF-PPP realize centimeter-level positioning. The difference between the results of two stations estimated with dual-frequency observations enhanced SF-PPP were compared with the relative positioning results computed with the DD algorithm. Experimental results showed that the relative positioning accuracy of the DD algorithm is slightly better than that of the dual-frequency observations enhanced SF-PPP. This could be explained by the effect of the float ambiguity resolutions on the positioning accuracy. The data was processed with the proposed method for the introduction of the DD ambiguity into SF-PPP and the results indicated that this method could improve the positioning accuracy and shorten the convergence time of the SF-PPP. The results could further improve the deformation monitoring ability of SF-PPP.

Molecular cloning and the expression pattern of AePOPB involved in the α-amanitin biosynthesis in Amanita exitialis fruiting bodies.

Amanita exitialis Zhu L. Yang & T. H. Li is the species responsible for the largest number of mushroom-associated human poisonings and fatalities in South China due to its lethal cyclic peptide toxins. Prolyl oligopeptidase B (POPB) is considered a key enzyme in the production of the highly toxic cyclic peptide α-amanitin. However, the POPB gene of A. exitialis has not been studied. In the present study we cloned and sequenced the full-length A. exitialis POPB (AePOPB) gene. The aim was to verify the gene structure and functions of AePOPB. The full-length sequence of AePOPB is 3144 bp, including 18 exons encoding 730 aa, and the advanced structure is very similar to that of the previously reported POPB in Galerina marginata (GmPOPB). The amino acid sequence of AePOPB is highly homologous with those from other amanitin-producing lethal mushrooms, implying that AePOPB may have a similar role in the biosynthesis of cyclic peptide toxins. Expression levels of AePOPB were detectable in all parts and developmental stages of the fruiting bodies, and AePOPB was expressed more strongly at early development stages (early and late elongation stages). At early and late elongation stages, the expression peaks occurred in the stipe, whereas at early and late mature stages, the expression peaks occurred in the pileus. The expression patterns of AePOPB in different stages and different parts of the fruiting bodies were highly consistent with those of Aeα-AMA, which is required for α-amanitin accumulation. These results indicate that AePOPB should be involved in the α-amanitin biosynthesis in A. exitialis.