Objective To investigate the immune response of IL-10 to Schistosoma japonicum infection in the early infectioin model and SEA immunization model of the IGTP~(-/-) and IRG-47~(-/-) mice.Methods In the early infection model,the IGTP knock out (IGTP~(-/-)) mice,IRG-47 knock out (IRG-47~(-/-)) mice and wild-type (WT) C57BL/6J mice were exposed to 300 S.japonicum cercariae via the pinna and sacrificed on day 7 post-infection.Each mouse pinna section was stained with hematoxylin-eosin (HE) to detect the pathological lesions,and the culture supernatant of pinna was used to test the levels of Th1/Th2 cytokines by indirect ELISA.In the SEA immunization model,IGTP~(-/-) IRG-47~(-/-) and WT mice were immunized with SEA twice and sacrificed in 3 weeks after the initial immunization.SEA-specific IgG antibody in sera was detected by indirect ELISA;the levels of Th1/Th2 cytokines were tested in culture supernatant of splenocytes by indirect ELISA;the proportions of CD4~+ T cells,CD8~+ T cells,B cells,Th1 and Th2 cells in the spleen were assayed by FACS.Results Although no obvious differences on the pathology of pinna were observed among the three mouse groups,the level of IL-10 in the culture supernatant of pinna of IRG-47~(-/-) mice was lower than that of IGTP~(-/-) mice in 7 days after the exposure.Following SEA immunization,the level of SEA-specific IgG antibody in sera of IGTP~(-/-) mice was lower than that in WT mice,the level of IL-10 in the culture supernatant of splenocytes of IRG-47~(-/-) mice was higher than that of IGTP~(-/-) and WT mice with the stimulation of SEA.However,the proportion of Th2 cells in the spleen of IRG47~(-/-) mice was the lowest among the three mouse groups.Conclusions SEA is the stimulus of IRG-47 deficiency mice to defend Schistosoma japanicum infection and promote the host to produce a protective response,and IL-10 may play an important role in immune regulation in this process.

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.

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

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.