Objective To explore the clinical feature of severe enterovirus 71 (EV71) associated hand foot and mouth disease (HFMD),and genotype of EV71.Methods Fluorescent quantitation PCR was done for detecting EV71.RT-PCR was performed to amplify VP1 for sequencing and identifying genotype.A retrospective analysis was performed based on the clinical data of 15 cases with severe EV71 infection.Results EV71 nucleotide was positive in all 15 cases.The genotype of EV71 was C4.All cases had abnormal temperature and followed with nervous symptoms in the early stage.Average time was 1.26 days from onset to severe complications appearance.Eleven cases progressed to neurogenic pulmonary edema.Four cases accepted nasal continuous positive airway pressure.Eleven cases accepted oral trachea cannula mechanical ventilation.Except for 3 cases died,one case abandoned,others 11 cases were cured.Conclusion The isolated strains of EV71 in this study are all C4 genotype.All cases with severe EV71 infection were followed with nervous symptoms in the early stage,most of whom would progress to neurogenic pulmonary edema.The mortality would be cut down by using mechanical ventilation in early stage.

β-thalassemia is caused by β-globin gene mutations.However,heterogeneous phenotypes were found in individuals with same genotype,and still undescribed mechanism underlies such variation.We collected blood samples from 30 β-thalassemia major,30 β-thalassemia minor patients,and 30 matched normal controls.Human lncRNA Array v2.0 (8 × 60 K,Arraystar) was used to detect changes in long non-coding RNAs (lncRNAs) and mRNAs in three samples each from β-thalassemia major,β-thalassemia minor,and control groups.Compared with normal controls,1424 and 2045 lncRNAs were up-and downregulated,respectively,in β-thalassemia major patients,whereas 623 and 349 lncRNAs were up-and downregulated,respectively,in β-thalassemia minor patients.Compared with β-thalassemia minor group,1367 and 2356 lncRNAs were up-and downregulated,respectively,in β-thalassemia major group.We selected five lncRNAs that displayed altered expressions (DQ583499,X-inactive specific transcript (Xist),lincRNA-TPM1,MRFS16P,and lincRNA-RUNX2-2) and confirmed their expression levels in all samples using real-time polymerase chain reaction.Based on coding-non-coding gene co-expression network and gene ontology biological process analyses,several signaling pathways were associated with three common organ systems exhibiting β-thalassemia phenotypes:hematologic,skeletal,and hepatic systems.This study implicates that abnormal expression levels of lncRNAs and mRNA in β-thalassemia cases may be correlated with its various clinical phenotypes.

To prepare the polyclonal antibody against the 3D polymerase of foot and mouth disease virus (FMDV), the 3D polymerase gene of this virus was amplified by PCR and doubly digested with BamH I and Nde I. Then, it were cloned into expression vector pET-30a(+) to obtain the recombinant plasmid pET-3D and this plasmid was transformed to E.coli BL21(DE3) with induction by IPTG.The target protein was identified and purified with SDS+PAGE, and the inclusion bodies were extracted. The purified target protein was used as antigen to immunize New Zealand rabbits to prepare the polyclonal antibody against 3D polymerase of FMDV, which was then characterized by indirect ELISA and Western blotting. As demonstrated by SDS-PAGE, the target protein with a molecular weight at 46 ku was expressed. The polyclonal antibody showed high affinity and obvious specificity and its titer was above 1:8 000. This polyclonal antibody may lay a foundation for the further studies on the biological functions and epitopes of the 3D polymerase of FMDV.