Objective To isolate the Japanese encephalitis virus carried by Culex tritaeniorhynchus in Dongchuan District of Yunnan Province and analyze its molecular characteristics, so as to provide insights into the prevention and control of Japanese encephalitis in Yunnan Province. Methods Mosquito specimens were collected using mosquito-trapping lamps from pig farms in Batang Village and Xiaoxin Village, Dongchuan District, Kunming City, Yunnan Province in July 2016, and the mosquito species was identified according to the mosquito morphology. Then, 60 to 100 mosquitoes of each species served as a group and were ground. Baby hamster kidney-21 (BHK-21) cells and Aedes albopictus clone C6/36 cells were used for virus isolation, and positive isolates were identified using flavivirus primers. The positive isolates were amplified using reverse transcription polymerase chain reaction (RT-PCR) assay with 15 pairs of specific primers covering the full length of the genotype I Japanese encephalitis virus, and DNA sequence assembly was performed using the software SeqMan in the DNASTAR package. The obtained sequences were aligned with the complete sequences of 38 Japanese encephalitis virus downloaded from the GenBank with the software MegAlign, and the nucleotide and amino acid homology analyses of the obtained sequences were performed. The difference in amino acid sites was analyzed with the software GeneDoc, and phylogenetic trees were created based on the sequences of the coding region and E protein of the isolated Japanese encephalitis virus with the software Mega X. In addition, the secondary and tertiary structures of the E protein of the Japanese encephalitis virus were predicted using the online tool SOPMA and the software Swiss-Model. Results A total of 5 820 mosquitoes were collected and 3 843 Cx. tritaeniorhynchus (66.03%) were identified according to the mosquito morphology. A positive virus isolate, termed YNDC55-33, was isolated from Cx. tritaeniorhynchoides following batches of virus isolation from mosquito specimens, and cytopathic effect was observed following inoculation into BHK-21 and C6/36 cells. The YNDC55-33 virus isolate was successfully amplified with the flavivirus primes, and a long sequence containing 300 nucleotides was obtained. Following sequence alignment using the BLAST tool, the sequence of the YNDC55-33 virus isolate had high homology with that of the genotype I Japanese encephalitis virus. A long sequence with 10 845 nucleotides in length, which encoded 3 432 amino acids, was obtained by splicing the full sequence of the YNDC55-33 virus isolate. Phylogenetic analysis based on the whole-genome sequence and E gene sequence of the YNDC55-33 virus isolate showed that the new YNDC55-33 virus isolate was most closely related to the genotype I Guizhou isolate (GenBank accession number: HM366552), with nucleotide homology of 98.5% and amino acid homology of 99.4%, and the YNDC55-33 virus isolate shared 97.96% ± 0.33% nucleotide homology and 99.35% ± 0.08% amino acid homology with other genotype I Japanese encephalitis virus isolates, and < 90% nucleotide homology and < 98% amino acid homology with other genotypes of Japanese encephalitis virus. The YNDC55-33 virus isolate and the live attenuated virus vaccine candidate SA14-14-2 isolate differed at 16 amino acid sites on E gene, and 7 out of 8 key amino acid sites related to neurovirulence. The secondary and tertiary structures of the E protein of the YNDC55-33 virus isolate were predicted to be characterized by random coils. Conclusions A genotype I Japanese encephalitis virus was isolated from Cx. tritaeniorhynchus in Dongchuan District, Kunming City. This virus isolate and the live attenuated virus vaccine candidate SA14-14-2 isolate does not differ at antigenic epitopes-related key amino acid sites, and the major protein structure of the virus isolate is random coils. This study adds new data for the epidemiological distribution of Japanese encephalitis virus in Yunnan Province, which may provide insights into the prevention and control of Japanese encephalitis in the province.

In the all-media era,online public opinion events are causing more and more trouble to the healthcare industry,and public hospitals in this situation are facing a severe test in terms of online public opinion management,which is generally problematic.It introduces the hospital network public opinion governance strategies and methods based on hazard vulnerability analysis of Peking University People's Hospital and explores a practical and feasible hos-pital online public opinion management model from the aspects of public opinion risk identification,public opinion management system construction,and improvement of departmental management level,to enhance the level of hospital public opinion governance,with a view to reducing and avoiding negative impacts on the hospital caused by online public opinion events,building a harmonious doctor-patient relationship,and improving the work of medical services.The aim is to reduce and avoid the negative impact of online public opinion events on the hospital,to build a harmonious doctor-patient relationship and improve medical services.

In the all-media era,online public opinion events are causing more and more trouble to the healthcare industry,and public hospitals in this situation are facing a severe test in terms of online public opinion management,which is generally problematic.It introduces the hospital network public opinion governance strategies and methods based on hazard vulnerability analysis of Peking University People's Hospital and explores a practical and feasible hos-pital online public opinion management model from the aspects of public opinion risk identification,public opinion management system construction,and improvement of departmental management level,to enhance the level of hospital public opinion governance,with a view to reducing and avoiding negative impacts on the hospital caused by online public opinion events,building a harmonious doctor-patient relationship,and improving the work of medical services.The aim is to reduce and avoid the negative impact of online public opinion events on the hospital,to build a harmonious doctor-patient relationship and improve medical services.

In the all-media era,online public opinion events are causing more and more trouble to the healthcare industry,and public hospitals in this situation are facing a severe test in terms of online public opinion management,which is generally problematic.It introduces the hospital network public opinion governance strategies and methods based on hazard vulnerability analysis of Peking University People's Hospital and explores a practical and feasible hos-pital online public opinion management model from the aspects of public opinion risk identification,public opinion management system construction,and improvement of departmental management level,to enhance the level of hospital public opinion governance,with a view to reducing and avoiding negative impacts on the hospital caused by online public opinion events,building a harmonious doctor-patient relationship,and improving the work of medical services.The aim is to reduce and avoid the negative impact of online public opinion events on the hospital,to build a harmonious doctor-patient relationship and improve medical services.

In the all-media era,online public opinion events are causing more and more trouble to the healthcare industry,and public hospitals in this situation are facing a severe test in terms of online public opinion management,which is generally problematic.It introduces the hospital network public opinion governance strategies and methods based on hazard vulnerability analysis of Peking University People's Hospital and explores a practical and feasible hos-pital online public opinion management model from the aspects of public opinion risk identification,public opinion management system construction,and improvement of departmental management level,to enhance the level of hospital public opinion governance,with a view to reducing and avoiding negative impacts on the hospital caused by online public opinion events,building a harmonious doctor-patient relationship,and improving the work of medical services.The aim is to reduce and avoid the negative impact of online public opinion events on the hospital,to build a harmonious doctor-patient relationship and improve medical services.

In the all-media era,online public opinion events are causing more and more trouble to the healthcare industry,and public hospitals in this situation are facing a severe test in terms of online public opinion management,which is generally problematic.It introduces the hospital network public opinion governance strategies and methods based on hazard vulnerability analysis of Peking University People's Hospital and explores a practical and feasible hos-pital online public opinion management model from the aspects of public opinion risk identification,public opinion management system construction,and improvement of departmental management level,to enhance the level of hospital public opinion governance,with a view to reducing and avoiding negative impacts on the hospital caused by online public opinion events,building a harmonious doctor-patient relationship,and improving the work of medical services.The aim is to reduce and avoid the negative impact of online public opinion events on the hospital,to build a harmonious doctor-patient relationship and improve medical services.

In the all-media era,online public opinion events are causing more and more trouble to the healthcare industry,and public hospitals in this situation are facing a severe test in terms of online public opinion management,which is generally problematic.It introduces the hospital network public opinion governance strategies and methods based on hazard vulnerability analysis of Peking University People's Hospital and explores a practical and feasible hos-pital online public opinion management model from the aspects of public opinion risk identification,public opinion management system construction,and improvement of departmental management level,to enhance the level of hospital public opinion governance,with a view to reducing and avoiding negative impacts on the hospital caused by online public opinion events,building a harmonious doctor-patient relationship,and improving the work of medical services.The aim is to reduce and avoid the negative impact of online public opinion events on the hospital,to build a harmonious doctor-patient relationship and improve medical services.

In the all-media era,online public opinion events are causing more and more trouble to the healthcare industry,and public hospitals in this situation are facing a severe test in terms of online public opinion management,which is generally problematic.It introduces the hospital network public opinion governance strategies and methods based on hazard vulnerability analysis of Peking University People's Hospital and explores a practical and feasible hos-pital online public opinion management model from the aspects of public opinion risk identification,public opinion management system construction,and improvement of departmental management level,to enhance the level of hospital public opinion governance,with a view to reducing and avoiding negative impacts on the hospital caused by online public opinion events,building a harmonious doctor-patient relationship,and improving the work of medical services.The aim is to reduce and avoid the negative impact of online public opinion events on the hospital,to build a harmonious doctor-patient relationship and improve medical services.

In the all-media era,online public opinion events are causing more and more trouble to the healthcare industry,and public hospitals in this situation are facing a severe test in terms of online public opinion management,which is generally problematic.It introduces the hospital network public opinion governance strategies and methods based on hazard vulnerability analysis of Peking University People's Hospital and explores a practical and feasible hos-pital online public opinion management model from the aspects of public opinion risk identification,public opinion management system construction,and improvement of departmental management level,to enhance the level of hospital public opinion governance,with a view to reducing and avoiding negative impacts on the hospital caused by online public opinion events,building a harmonious doctor-patient relationship,and improving the work of medical services.The aim is to reduce and avoid the negative impact of online public opinion events on the hospital,to build a harmonious doctor-patient relationship and improve medical services.

Objective To investigate the effect of glycine N-methyl transferase (GNMT)on intrauterine adhesion (IUA)fibrosis and its related mechanism.Methods In vivo experiment:A total of 36 healthy female SD rats (SPF grade,6～8 weeks old and weighing from 180～220 g)were subjected in this study.IUA model of SD rats and IUA model of GNMT overexpressed rats were established.RT-qPCR and immunofluorescence assay were applied to detect GNMT expression level in normal uterus and model group.RT-qPCR and Western blotting were used to detect the mRNA and protein levels of fibrosis-related molecules and the activation of TGF-β1/Smad3 signaling pathway in each group.The number of endometrial glands in each group was observed by HE staining.Masson staining was used to analyze the severity of endometrial fibrosis in each group.In vitro experiment:transformed human endometrial stromal cells (THESCs)fibrotic phenotype model was constructed using TGF-β1,and THESCs stably transfected with GNMT overexpression lentvirus were treated with TGF-β1.RT-qPCR and Western blotting were used to detect the mRNA and protein expression of fibrosis-related molecules.The expression of TGF-β1/Smad3 signaling pathway was detected by Western blotting.TGF-β1/Smad3 signaling pathway was activated by TGF-β1/Smad signaling pathway activator (SRI-011381),and the expression of TGF-β1/Smad3 signaling pathway and key molecular proteins of fibrosis phenotype was measured with Western blotting.Results In vivo experiment,the mRNA and protein expression levels of GNMT were significantly decreased in the IUA rats than the control rats (P<0.05).Overexpression of GNMT decreased the mRNA and protein levels of fibrosis related molecules,Collagen Ⅰ,Collagen Ⅲ and FN in the IUA rats (P<0.05),and decreased the phosphorylation levels of TGF-β1 and its downstream Smad3 protein (P<0.05).HE and Masson staining showed that overexpression of GNMT could increase the number of endometrial glands and reduce the severity of fibrosis in the IUA rats (P<0.05).In vitro experiments:overexpression of GNMT decreased the mRNA and protein levels of Collagen Ⅰ,Collagen Ⅲ and FN associated with fibrotic phenotype of THESCs (P<0.05),and reduced the phosphorylation level of Smad3 protein,downstream of TGF-β1 (P<0.05).After activation of TGF-β1/Smad3 signaling pathway,the protein levels of TGF-β1/Smad3 signaling pathway and downstream fibrosis phenotype molecules,Collagen Ⅲ and FN,were significantly decreased in the LV-GNMT+SRI-011381 group.Conclusion Overexpression of GNMT can inhibit endometrial fibrosis by regulating TGF-β1/Smad3 signaling pathway,thus achieving therapeutic effect on IUA.