Global quantitative proteomic analysis profiles of host protein expression in response to enterovirus A71 infection in bronchial epithelial cells based on tandem mass tag (TMT) peptide labeling coupled with LC-MS/MS uncovers the key role of proteasome in virus replication.

Enterovirus A71 (EV-A71) is a neurotropic human pathogen which mainly caused hand, foot and mouth disease (HFMD) mostly in children under 5 years-old. Generally, EV-A71-associated HFMD is a relatively self-limiting febrile disease, but there will still be a small percentage of patients with rapid disease progression and severe neurological complications. To date, the underlying mechanism of EV-A71 inducing pathological injury of central nervous system (CNS) remains largely unclear. It has been investigated and discussed the changes of mRNA, miRNA and circRNA expression profile during infection by EV-A71 in our previous studies. However, these studies were only analyzed at the RNA level, not at the protein level. It's the protein levels that ultimately do the work in the body. Here, to address this, we performed a tandem mass tag (TMT) peptide labeling coupled with LC-MS/MS approach to quantitatively identify cellular proteome changes at 24 h post-infection (hpi) in EV-A71-infected 16HBE cells. In total, 6615 proteins were identified by using TMT coupled with LC-MS/MS in this study. In the EV-A71- and mock-infected groups, 210 differentially expressed proteins were found, including 86 upregulated and 124 downregulated proteins, at 24 hpi. To ensure the validity and reliability of the proteomics data, 3 randomly selected proteins were verified by Western blot and Immunofluorescence analysis, and the results were consistent with the TMT results. Subsequently, functional enrichment analysis indicated that the up-regulated and down-regulated proteins were individually involved in various biological processes and signaling pathways, including metabolic process, AMPK signaling pathway, Neurotrophin signaling pathway, Viral myocarditis, GABAergic synapse, and so on. Moreover, among these enriched functional analysis, the "Proteasome" pathway was up-regulated, which has caught our attention. Inhibition of proteasome was found to obviously suppress the EV-A71 replication. Finally, further in-depth analysis revealed that these differentially expressed proteins contained distinct domains and localized in different subcellular components. Taken together, our data provided a comprehensive view of host cell response to EV-A71 and identified host proteins may lead to better understanding of the pathogenic mechanisms and host responses to EV-A71 infection, and also to the identification of new therapeutic targets for EV-A71 infection.

Application of alpha-enolase combined with CYFRA21-1 and CA125 in diagnosis of malignant pleural effu-sion / 实用医学杂志

Objective To compare the concentrations of alpha-enolase (ENO1),CYFRA21-1,and CA125 in the patients with malignant pleural effusion ,tuberculous exudative pleural effusion ,or parapneumonia pleural effusion. To explore the clinical value of ENO1 in pleural effusion combined with serum CYFRA21-1 and CA125 in diagnosis of malignant pleural effusion. Methods Enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of ENO1 in pleural effusions. The concentrations of CA125 and CYFRA21-1 in the blood samples were measured using chemiluminescence and magnetic particle-based chemiluminescence respective-ly. The sensitivity and specificity of ENO1 combined with serum CYFRA21-1 and CA125 detection were calculated. Results The concentration of ENO1 in malignant pleural effusion group was significantly increased(P<0.001);the concentrations of ENO1 did not differ significantly between tuberculous exudative pleural effusion and parapneu-monia pleural effusion(P>0.05). The sensitivity and specificity of ENO1 combined with serum CYFRA21-1 and CA125 detection in malignant pleural effusion were 94％ and 74％,98％ and 98％,respectively. Conclusions ENO1 combined with serum CYFRA21-1 and CA125 detection can improve the sensitivity of diagnosis of malignant pleural effusion and enhance the diagnostic rate of malignant pleural effusion.

Application of alpha-enolase combined with CYFRA21-1 and CA125 in diagnosis of malignant pleural effu-sion / 实用医学杂志

Objective To compare the concentrations of alpha-enolase (ENO1),CYFRA21-1,and CA125 in the patients with malignant pleural effusion ,tuberculous exudative pleural effusion ,or parapneumonia pleural effusion. To explore the clinical value of ENO1 in pleural effusion combined with serum CYFRA21-1 and CA125 in diagnosis of malignant pleural effusion. Methods Enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of ENO1 in pleural effusions. The concentrations of CA125 and CYFRA21-1 in the blood samples were measured using chemiluminescence and magnetic particle-based chemiluminescence respective-ly. The sensitivity and specificity of ENO1 combined with serum CYFRA21-1 and CA125 detection were calculated. Results The concentration of ENO1 in malignant pleural effusion group was significantly increased(P<0.001);the concentrations of ENO1 did not differ significantly between tuberculous exudative pleural effusion and parapneu-monia pleural effusion(P>0.05). The sensitivity and specificity of ENO1 combined with serum CYFRA21-1 and CA125 detection in malignant pleural effusion were 94％ and 74％,98％ and 98％,respectively. Conclusions ENO1 combined with serum CYFRA21-1 and CA125 detection can improve the sensitivity of diagnosis of malignant pleural effusion and enhance the diagnostic rate of malignant pleural effusion.