ABSTRACT
ABSTRACT New viruses of the Picornavirales order have been discovered with the increase in the number of sequences obtained by high-throughput sequencing, as well as human stool-associated RNA virus (husavirus [HuV]), found in human stool samples. However, there is much to be clarified about HuV. Its cellular host, evolutionary history, and other biological characteristics are still unknown. Therefore, samples collected from human beings and environmental samples in a watershed in Southern Brazil were processed for the metagenomic library. Upon metagenomic analysis, we identified a HuV (husavirus LMM_67754 OP019707) genome with 8,846 bp, which was reported for the first time in Southern Brazil. The new genome presents only 37% of nucleotide identity with Brazilian strains and more than 90% with genomes from China, Vietnam, Venezuela, and the Netherlands. The HuV phylogeny presents significant differences among genomes, probably because multiple introductions of the virus may have occurred. Many questions still need to be answered about HuV. Therefore, more sequences and studies on this virus are necessary to improve the comprehension of the unknown origin of Picornavirales.
ABSTRACT
; Aim To study the synergistic inhibition effect of hydroxychloroquine (HCQ) and bevacizumab (B E V) on the cell growth of HUVECs and its mechanism. Methods RTCA,flow apoptosis assay and the vascularization assay of HUVECs were used to study the effect of HCQ combined with BEV on HUVEC proliferation, cell apoptosis and vascularization. The expressions of LC3 and p62 proteins were detected by Western blot, autophagy flow was observed through m Cherry EGFPLC3 double fluorescence, the changes of autophagosome and mitochondria were observed by electron microscopy, and the regulatory molecular mechanism of HCQ combined with BEV was revealed by transcriptome sequencing (RNA-seq). Results Compared with single drug groups, the combination of HCQ and BEV could synergistically inhibit HUVEC proliferation, promote HUVEC apoptosis, and inhibit the formation of vascular structures in HUVECs. HCQ could inhibit autophagy of HUVECs. Transcriptome sequencing results showed that the combination of HCQ and BEV influenced 69 different expressed genes,and the combination of HCQ and BEV might synergistically inhibit HUVECs by regulating "hub" genes such as RICTOR and PIK3CA. Conclusions The combination of HCQ and BEV can synergistically inhibit HUVECs, which is related to not only the inhibition of autophagy by HCQ, but also the regulation of " hub " genes such as RICTOR and PIK3CA.