This study was aimed to examine the effect of TREK-1 silencing on the function of astrocytes. Three 21-nucleotide small interfering RNA (siRNA) duplexes (siT1, siT2, siT3) targeting TREK-1 were constructed. Cy3-labeled dsRNA oligmers were used to determine the transfection efficiency in cultured astrocytes. TREK-1-specific siRNA duplexes (siT1, siT2, siT3) at the optimal concentration were transfected into cultured astrocytes, and the most efficient siRNA was identified by the method of immunocytochemical staining and Western blotting. The proliferation of astrocytes tranfected with TREK-1-targeting siRNA under hypoxia condition was measured by fluorescence-activated cell sorting (FACS). The results showed that TREK-1 was expressed in cultured astrocytes. The dsRNA oligmers targeting TREK-1 could be transfected efficiently in cultured astrocytes and down-regulate the expression of TREK-1 in astrocytes. Moreover, the down-regulation of TREK-1 in astrocytes contributed to the proliferation of astrocytes under hypoxia condition as determined by cell cycle analysis. It was concluded that siRNA is a powerful technique that can be used to knockdown the expression of TREK-1 in astrocytes, which helps further investigate the function of TREK-1 channel in astrocytes under physicological and pathological condition.

OBJECTIVETo investigate the expression of the peripheral blood retinoid-related orphan receptor gamma t (RORgamma t) and 4-1BB/4-1BBL mRNA in oral lichen planus (OLP) patients.

METHODSThe expression levels of peripheral blood' RORyt mRNA and 4-1BB/4-1BBL mRNA of 30 samples of OLP patients and 30 cases of healthy human were detected using real-time fluorescent quantitative-polymerase chain reaction (FQ-PCR) technology.

RESULTSIn the OLP group, the expression level of RORgamma t, 4-1BB, 4-1BBL mRNA were higher than those in the control group (P < 0.05). After the conversion of the 2-delta delta CT, the expression levels of three kinds of mRNA in the OLP group were 3.087, 3.320 and 4.005 times of the control group respectively. In the OLP group, the expression levels of RORgamma t mRNA and 4-1BB mRNA had no obvious correlation (P > 0.05), the expression levels of 4-1BB and 4-1BBL mRNA had a positive correlation (r = 0.455 0, P < 0.05).

CONCLUSIONThe abnormal expression of RORyt, 4-1BB and 4-1BBL mRNA may play certain roles in the pathogenesis of OLP.

Humans ; Lichen Planus, Oral ; Male ; RNA, Messenger ; Retinoids

This study was aimed to examine the effect of TREK-1 silencing on the function of astrocytes. Three 21-nucleotide small interfering RNA (siRNA) duplexes (siT1, siT2, siT3) targeting TREK-1 were constructed. Cy3-labeled dsRNA oligmers were used to determine the transfection efficiency in cultured astrocytes. TREK-1-specific siRNA duplexes (siT1, siT2, siT3) at the optimal concentration were transfected into cultured astrocytes, and the most efficient siRNA was identified by the method of immunocytochemical staining and Western blotting. The proliferation of astrocytes tranfected with TREK-1-targeting siRNA under hypoxia condition was measured by fluorescence-activated cell sorting (FACS). The results showed that TREK-1 was expressed in cultured astrocytes. The dsRNA oligmers targeting TREK-1 could be transfected efficiently in cultured astrocytes and down-regulate the expression of TREK-1 in astrocytes. Moreover, the down-regulation of TREK-1 in astrocytes contributed to the proliferation of astrocytes under hypoxia condition as determined by cell cycle analysis. It was concluded that siRNA is a powerful technique that can be used to knockdown the expression of TREK-1 in astrocytes, which helps further investigate the function of TREK-1 channel in astrocytes under physicological and pathological condition.
Animals ; Astrocytes ; physiology ; Cells, Cultured ; Gene Silencing ; physiology ; Potassium Channels ; Potassium Channels, Tandem Pore Domain ; genetics ; RNA Interference ; physiology ; RNA, Small Interfering ; genetics ; Rats

OBJECTIVE： To analyze and predict potential pharmacological effects and mechanism of flavonoids from Glycyrrhiza uralensis. METHODS： By the means of network pharmacology， according to TCM integrative pharmacology platform （TCMSP）， using oral bioavailability （OB）＞30％ and drug-likeness （DL）＞0.18 of compound as reference， flavonoids compound of G. uralensis were screened. The potential targets of flavonoids were predicted with pharmacophore matching and PharmMapper date base. DAVID V 6.8 analysis tool was used for KEGG signaling pathway analysis and GO biological process enrichment analysis （using P＜0.05 as judgement standard） of target protein. A flavonoids-targets-signaling pathways network was built through Cytoscape 3.5.1 software. RESULTS： A total of 19 flavonoids compounds （such as liquiritin， isoliquiritin and liquiritigenin， etc.） were screened， involving 78 target proteins as cellular retinoic acid-binding protein 2 and neprilysin （188 times in total）， 40 signaling pathways （among them， 8 pathways related to cancer， 8 pathways related to endocrine system， 6 pathways related to signal transduction， 5 pathways related to infectious diseases and 3 pathways related to metabolism） as insulin signaling pathway， PI3K-Akt and so on. The flavonoids-targets-signaling pathways network model showed that flavonoids compounds of G. uralensis could act on different metabolic pathways through multiple targets. CONCLUSIONS： The flavonoids of G. uralensis have therapeutic effect on diseases of cancer， endocrine system， infectious diseases， metabolism and so on. It may have potential anti-parkinson’s effect.

Enterotoxigenic Escherichia coli（ETEC）infection can induce watery diarrhea，leading to dehydration，electrolyte disturbance，and even death in severe cases. Recombinant B subunit/inactivated whole-cell cholera（rBS/WC）vaccine is effective in preventing ETEC infectious diarrhea. On the basis of the latest evidence on etiology and epidemiology of ETEC，as well as the effectiveness，safety，and health economics of rBS/WC vaccine，National Clinical Research Center for Child Health（The Children’s Hospital，Zhejiang University School of Medicine）and Zhejiang Provincial Center for Disease Control and Prevention invited experts to develop expert consensus on rBS/WC vaccine in prevention of ETEC infectious diarrhea. It aims to provide the clinicians and vaccination professionals with guidelines on using rBS/WC vaccine to reduce the incidence of ETEC infectious diarrhea.