Porcine hemagglutinating encephalomyelitis virus(PHEV)is highly prevalent and wide-ly distributed,and its harm on pig farming is of great concern,therefore,effective strategies for prevention and control are necessary.In this study,chloroquine(CQ)was selected and its effect and mechanism on replication and proliferation of PHEV were investigated.The results showed that CQ(1-100 mmol/L)inhibits the replication and proliferation of PHEV in a dose-dependent manner and has no cytotoxicity on N2a cells.The production of inflammatory cytokines and activa-tion of NF-κB signaling pathway were both inhibited by CQ concentration at 50 mmol/L in PHEV-infected N2a cells.In conclusion,we confirmed the inhibition of CQ in replication and proliferation of PHEV.It is expected to be applied to the prevention and treatment of PHEV in clinical.

Porcine hemagglutinating encephalomyelitis virus(PHEV)is one of the coronaviruses susceptible to swine populations.The non-structural protein 2(NS2)encoded by its genome is fre-quently deleted during the epidemic transmission of the virus,but its biological significance re-mains unclear.In order to explore the structure and function of the NS2 protein,this study utilized platforms such as ProtParam,TMHMM,NetPhos3.1,and ExPASy to analyze its physicochemical properties,spatial structure,genetic evolution,and post-translational modification characteristics.Meanwhile,the NS2 protein was expressed in eukaryotes and transcriptome sequencing was per-formed to clarify the biological processes it participates in.The results showed that the NS2 protein consists of 233 amino acids,with a molecular weight of 26.735 kDa,and a half-life of approximately 30 hours in mammals.It includes 13 phosphorylation sites,2 N-glycosylation sites,and 1 O-glyco-sylation site,with no signal peptide and strong hydrophilicity.The a-helix accounts for the highest proportion in NS2(43.78％),followed by random coils(36.05％).The homology of the NS2 pro-tein between the epidemic strains PHEV-CC14 and PHEV-JL/2008 in Northeast China is 99.57％.The NS2 protein is widely involved in the regulation of nerve-related functions,such as axon guid-ance and synaptic development.This study preliminarily clarified the biological function of the NS2 protein,providing a new perspective for understanding the pathogenic mechanism of PHEV.

Porcine hemagglutinating encephalomyelitis virus(PHEV)is one of the coronaviruses susceptible to swine populations.The non-structural protein 2(NS2)encoded by its genome is fre-quently deleted during the epidemic transmission of the virus,but its biological significance re-mains unclear.In order to explore the structure and function of the NS2 protein,this study utilized platforms such as ProtParam,TMHMM,NetPhos3.1,and ExPASy to analyze its physicochemical properties,spatial structure,genetic evolution,and post-translational modification characteristics.Meanwhile,the NS2 protein was expressed in eukaryotes and transcriptome sequencing was per-formed to clarify the biological processes it participates in.The results showed that the NS2 protein consists of 233 amino acids,with a molecular weight of 26.735 kDa,and a half-life of approximately 30 hours in mammals.It includes 13 phosphorylation sites,2 N-glycosylation sites,and 1 O-glyco-sylation site,with no signal peptide and strong hydrophilicity.The a-helix accounts for the highest proportion in NS2(43.78％),followed by random coils(36.05％).The homology of the NS2 pro-tein between the epidemic strains PHEV-CC14 and PHEV-JL/2008 in Northeast China is 99.57％.The NS2 protein is widely involved in the regulation of nerve-related functions,such as axon guid-ance and synaptic development.This study preliminarily clarified the biological function of the NS2 protein,providing a new perspective for understanding the pathogenic mechanism of PHEV.

Porcine hemagglutinating encephalomyelitis virus(PHEV)is highly prevalent and wide-ly distributed,and its harm on pig farming is of great concern,therefore,effective strategies for prevention and control are necessary.In this study,chloroquine(CQ)was selected and its effect and mechanism on replication and proliferation of PHEV were investigated.The results showed that CQ(1-100 mmol/L)inhibits the replication and proliferation of PHEV in a dose-dependent manner and has no cytotoxicity on N2a cells.The production of inflammatory cytokines and activa-tion of NF-κB signaling pathway were both inhibited by CQ concentration at 50 mmol/L in PHEV-infected N2a cells.In conclusion,we confirmed the inhibition of CQ in replication and proliferation of PHEV.It is expected to be applied to the prevention and treatment of PHEV in clinical.