AIM:To explore the effect of tomatidine(TA)on lipopolysaccharide(LPS)-induced nerve cell in-jury and the underlying mechanism.METHODS:The neuroinflammation model was induced by treating SH-SY5Y cells with LPS.These cells were divided into control(CON),LPS,and LPS+TA groups.The LPS group was treated with 5 μg/mL LPS for 24 h to establish an inflammatory model.The LPS+TA group was first treated with 5 μmol/L tomatidine for 24 h and then co-cultured with 5 μg/mL LPS for 24 h.Cell viability was detected using the CCK-8 assay.RT-qPCR was used to detect the mRNA expression of inflammatory factors tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β).The protein expression of transcription factor EB(TFEB),p-TFEB,P62,and microtubule-associated protein 1 light chain 3(LC3)expression was detected through Western blot.TFEB localization and cleaved caspase-3 expression were detected through immunofluorescence.The cell apoptosis rate was detected through flow cytometry.RESULTS:(1)Compared with the CON group,the LPS group exhibited significant increases in IL-1β and TNF-α mRNA levels(P＜0.05),the cell apoptosis rate,and the p-TFEB level(P＜0.01).By contrast,P62,LC3-Ⅱ/LC3-Ⅰ,and TFEB protein ex-pression levels decreased significantly(P＜0.05),and TFEB was mainly localized in the cytoplasm.(2)Compared with the LPS group,tomatidine treatment significantly decreased the p-TFEB protein expression level(P＜0.01),increased the TFEB protein expression level(P＜0.01),and promoted the TFEB protein to migrate into the nucleus.After treatment of tomatidine,the LC3-Ⅱ/LC3-Ⅰ protein expression level significantly increased(P＜0.05),and the cell apoptosis rate signifi-cantly decreased(P＜0.01).In addition,the TNF-α mRNA level significantly decreased after tomatidine treatment(P＜0.01).CONCLUSION:Tomatidine improves autophagy dysfunction,inflammatory reaction,and cell apoptosis induced by LPS via activating the transcription factor EB.

OBJECTIVETo investigate pathogens and molecular-epidemiology characteristics of viral meningoencephalitis in the monitoring sites of Zhejiang province, 2013.

METHODSCerebrospinal fluid and/or stool specimens were collected from suspected patients admitted to the monitoring hospitals in southern and northern Zhejiang province. Such specimen were subject to real-time qPCR for the detection of Human enterovirus (HEV), Japanese encephalitis virus (JEV), Mumps virus (MuV), Herpes simplex virus (HSV) and Cytomegalovirus (CMV). HEVs were isolated using the RD and Hep-2 cell lines, while VP1 genes from all HEV-positive isolates or RNA-positive specimen were amplified, sequenced, for homology and evolution analysis.

RESULTS92 (38.5%) of the 239 samples collected from 229 patients were detected as virus nucleic acid positive, including 87 HEV positive samples, 1 MuV positive, 2 HSV positive, and 2 CMV positive; of the 87 HEV positive samples, 38 were further determined to be Coxsackievirus (CV) and 49 as Echovirus (E). 56 HEV strains were isolated from 239 (23.4%) samples. From the 31 cerebral fluid specimen of nucleic acid positive yet virus isolation negative, the most specimen were identified with E9 (9 specimen), followed by CVA9 (8 specimen); the viral serotype of Zhejiang province HEV were CVA9, CVB4, CVB5, E6, E7, E9, E11, E14, E16, E25 and E30, respectively. Predominant epidemic strains identified at southern and northern Zhejiang province were CVB5 and E6 respectively. The phylogenetic analysis of VP1 gene showed that all the HEV isolates in Zhejiang province were HEV-B.

CONCLUSIONThe HEV-B was the main pathogen for viral meningoencephalitis in Zhejiang province in 2013, including 11 serotypes, while E7 was the first time to be isolated in Zhejiang province. The predominant isolates were CVB5 and E6 in southern and northern Zhejiang province respectively. The positive rate of viral nucleic acid detection was significantly higher than that of viral isolation. Regular EV isolation method was exposed to the risk of missing-detection of E9 and CVA9.

Biological Evolution ; China ; epidemiology ; Cytomegalovirus ; Encephalitis Virus, Japanese ; Encephalitis, Viral ; Enterovirus ; Enterovirus B, Human ; Hepatitis E virus ; Humans ; Meningitis, Viral ; epidemiology ; genetics ; Meningoencephalitis ; Molecular Epidemiology ; Mumps virus ; Phylogeny