Human astrovirus (HastV) is recognized as one of the leading causes of acute viral diarrhea in infants. The HastV non-structural protein, nsPla, and C-terminal protein, nsPla/4, contain various conserved functional domains,and may play an important role in virus replication, transcription and the virus-host interactions of HastV. This study used an E. coli system to investigate the expression of nsPla and nsPla/4 proteins. Firstly,the nsPla and nsPla/4 genes of HAstV-1 were cloned into the prokaryotic expression vector,PGEX-4T-1, to build the PGEX-4T-1a and PGEX-4T-la/4 fusion protein plasmids. Then, the recombinant plasmids were transformed into Escherichia coli BL21 (DE3) and induced with isopropyl-β-D-thiogalactopyranoside (IPTG). The optimal expression conditions of the two fusion proteins were identified and then analyzed by polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting, respectively. The results showed that the pGEX-4T-la fusion protein was maximally expressed at 30 °C after 12 hours of induction with 1.0 mM IPTG. The pGEX-4T-la/4 fusion protein was maximally expressed at 20 °C after 8 hours of induction with 0.5 mM IPTG. Western blot analysis showed that the two fusion proteins specificity reacted with the anti-nsPla and anti-GST monoclonal antibodies, respectively. This study successfully obtained the HAstV non-structural protein, nsP1a, and its C-terminal protein nsP1a/4 protein using an E. coli system. This novel study lays the foundation for future research into the pathogenic mechanisms of human astrovirus and the functions of its non-structural protein.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Mamastrovirus ; genetics ; metabolism ; Viral Nonstructural Proteins ; genetics ; metabolism

Objective:To explore the effect of time-sensitive incentive nursing on psychological resilience, quality of life, nursing satisfaction and disease cognition of postoperative patients with chronic sinusitis (CS) .Methods:From January 2019 to July 2020, the convenience sampling method was used to select 60 postoperative patients with CS who were admitted to Fuyang Hospital of Anhui Medical University as the research objects. Patients were randomly divided into the observation group and the control group, with 30 cases in each group. The control group carried out conventional nursing, and the observation group was given conventional nursing combined with time-sensitive incentive nursing. The Connor-Davidson Resilience Scale (CD-RISC) , the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) dimensions of mental health, general health, physical functioning and role emotional were used to compare the psychological resilience and quality of life of the two groups of patients before and after nursing. Disease cognition and nursing satisfaction of the two groups of patients after nursing were compared.Results:The scores of the CD-RISC dimensions, disease cognition, four dimensions of the SF-36, and nursing satisfaction of patients in the observation group after nursing were higher than those of the control group, and the differences were statistically significant ( P<0.01) . Conclusions:The application of time-sensitive incentive nursing in postoperative patients with CS can improve their psychological resilience, disease cognition, quality of life and nursing satisfaction.