Two human Fab antibodies against avian influenza A (H5N1) virus were obtained by panning a H5N1 patient-derived antibody phage library using purified virions of the H5N1 patient isolate A/Anhui/1/2005 and HA protein of the H5N1 reference viruse A/Viet Nam/1203/2004. After testing the binding properties and antiviral function to H5N1 virus, the selected Fab antibodies were converted to full human IgG antibodies with recombinant baculovirus/insect cell system. Both mAbs, AVFluIgG01 and AVFluIgG03, bound to HA in immunofluorescence assay (IFA) without cross-reaction with the other substypes of influenza A viruses (H1N1, H3N2). The cross-reactivity of the two antibodies for different strains of H5N1 was tested in vitro by micro-neutralization assays. In vitro, mAb AVFluIgG01 potently neutralized not only the selected well-characterized Clade 2 H5N1 viruses isolated from mainland of China except A/Guangdong/1/2006, but also the Clade 1 representative isolate A/Viet Nam/1203/2004; and AVFluIgG03 neutralized all the selected Clade 2 H5N1 viruses isolated from mainland of China, but had no neutralizing activity with the Clade 1 H5N1 virus A/Viet Nam/1203/2004. The results bring new prospect for the prophylaxis or treatment of H5N1 virus infection and may provide a clue for novel vaccine development.
Amino Acid Sequence ; Animals ; Antibodies, Viral ; genetics ; immunology ; Antibody Specificity ; Birds ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Influenza Vaccines ; genetics ; immunology ; Influenza in Birds ; immunology ; virology ; Molecular Sequence Data ; Neutralization Tests ; Recombinant Proteins ; immunology ; Sequence Homology, Amino Acid

Aim To investigate the protective effect of Huoxue Jiedu recipe on autophagy injury of H9C2 cardiomyocytes induced by hypoxia/reoxygenation and its mechanism. Methods H9C2 cardiomyocytes were used to establish a hypoxia/reoxygenation injury model. The effective concentration was screened and the cell activity was detected by CCK8 assay. The apoptotic rate of myocardial cells was detected by flow cytometry. The expression of autophagy marker LC3 was observed by laser confocal microscopy. The mRNA levels of Beclin-1, LC3 and Bcl-2 were detected by real-time quantitative PCR. The expressions of Beclin-1, LC311/I, Cleaved caspase-3, β-catenin, p-p65, Bcl-2, p62, p-Akt, p-mTOR were detected by Western blot. Results Huoxue Jiedu recipe can enhance the growth activity of myocardial cells and reduce the apoptotic rate and autophagy level, and it can enhance the activation of PI3K/Akt/mTORCl pathway, decrease Beclin-1 and LC3 mRNA levels, while increase Bcl-2 mRNA levels. It also decreased the expression of Beclin-1, LC311/I, Cleaved caspase-3, β-catenin, p-p65, and increased the expression of p62, p-Akt, p-mTOR, and Bcl-2. Conclusions Huoxue Jiedu recipe can reduce the level of autophagy and apoptosis of myocardial cells by regulating the autophagy pathway of PI3K/Akt/mTORCl, thereby playing a protective role in hypoxia/reoxygenation H9C2 myocardial cells.

This study aimed to investigate bacterial community in an urban drinking water distribution system (DWDS) during an occurrence of colored water. Variation in the bacterial community diversity and structure was observed among the different waters, with the predominance of Proteobacteria. While Verrucomicrobia was also a major phylum group in colored water. Limnobacter was the major genus group in colored water, but Undibacterium predominated in normal tap water. The coexistence of Limnobacter as well as Sediminibacterium and Aquabacterium might contribute to the formation of colored water.
Bacteria ; genetics ; isolation & purification ; Drinking Water ; microbiology ; RNA, Ribosomal, 16S ; genetics ; Water Microbiology ; Water Supply ; analysis