Objective To explore the effects and mechanism of Baohuoside Ⅰ on transcription factors forkhead box protein 3(FoxP3)in regulatory T cell(Treg).Methods Treat induced regulatory T cells(iTreg cells:0,2,5,10,15,20 μmol·L-1 Baohuoside Ⅰ;MT-2 cells:0,2,5,10,20 μmol·L-1 Baohuoside Ⅰ)with different concentrations of Baohuoside Ⅰ for 16 hours.Use flow cytometry to detect the expression level of FoxP3,and use Western Blot to detect the expression level of FoxP3 protein.MT-2 cells were further divided into a control group and an experimental group.The control group was treated with dimethyl sulfoxide(DMSO)and 10 μg·mL-1 cycloheximide(CHX)for 0,2,4 and 6 hours;the experimental group was treated with 20 μmol·L-1Baohuoside Ⅰ and 10 μ g·mL-1 CHX for 0,2,4 and 6 hours.The degradation rate of FoxP3 in MT-2 cells from the control and experimental groups was detected by flow cytometry at different time points.Results In iTregs cells,the expression levels of FoxP3 in 0,2,5,10,15 and 20 μmol·L-1 Baohuoside Ⅰ groups were 1.00±0.00,0.98±0.07,0.95±0.04,0.86±0.04,0.57±0.04 and 0.55±0.04;the relative expression levels of FoxP3 protein were 1.00±0.00,1.04±0.14,0.86±0.06,0.64±0.33,0.36±0.15 and 0.10±0.04.The differences of above indexes between the 10,15 and 20 μmol·L-1 Baohuoside Ⅰ groups and the 0 μmol·L-1 Baohuoside Ⅰ group were statistically significant(all P＜0.001).In MT-2s cells,the expression levels of FoxP3 in0,2,5,10 and 20 μmol·L-1 Baohuoside Ⅰ groups were 1.00±0.00,0.88±0.05,0.84±0.08,0.70±0.16 and 0.57±0.14;the relative expression levels of FoxP3 protein were 1.00±0.00,0.89±0.10,0.76±0.19,0.70±0.19 and 0.37±0.12.The differences of above indexes between the 5,10 and 20 μmol·L-1 Baohuoside Ⅰ groups and the 0 μmol·L-1 Baohuoside Ⅰ group were statistically significant(P＜0.01,P＜0.001).Furthermore,Baohuoside Ⅰ decreased the stability of FoxP3 which can be observed in the CHX pulse-chase assay at the 6 hour time point where the expression levels of FoxP3 in the control group at 0.83±0.06 compared to the experimental group at 0.55±0.11(P＜0.001).Conclusion Baohuoside Ⅰcan destabilize FoxP3 in Tregs which has been shown to reduce the immunosuppressive functions of Tregs.

Objective To explore the effects and mechanism of Baohuoside Ⅰ on transcription factors forkhead box protein 3(FoxP3)in regulatory T cell(Treg).Methods Treat induced regulatory T cells(iTreg cells:0,2,5,10,15,20 μmol·L-1 Baohuoside Ⅰ;MT-2 cells:0,2,5,10,20 μmol·L-1 Baohuoside Ⅰ)with different concentrations of Baohuoside Ⅰ for 16 hours.Use flow cytometry to detect the expression level of FoxP3,and use Western Blot to detect the expression level of FoxP3 protein.MT-2 cells were further divided into a control group and an experimental group.The control group was treated with dimethyl sulfoxide(DMSO)and 10 μg·mL-1 cycloheximide(CHX)for 0,2,4 and 6 hours;the experimental group was treated with 20 μmol·L-1Baohuoside Ⅰ and 10 μ g·mL-1 CHX for 0,2,4 and 6 hours.The degradation rate of FoxP3 in MT-2 cells from the control and experimental groups was detected by flow cytometry at different time points.Results In iTregs cells,the expression levels of FoxP3 in 0,2,5,10,15 and 20 μmol·L-1 Baohuoside Ⅰ groups were 1.00±0.00,0.98±0.07,0.95±0.04,0.86±0.04,0.57±0.04 and 0.55±0.04;the relative expression levels of FoxP3 protein were 1.00±0.00,1.04±0.14,0.86±0.06,0.64±0.33,0.36±0.15 and 0.10±0.04.The differences of above indexes between the 10,15 and 20 μmol·L-1 Baohuoside Ⅰ groups and the 0 μmol·L-1 Baohuoside Ⅰ group were statistically significant(all P＜0.001).In MT-2s cells,the expression levels of FoxP3 in0,2,5,10 and 20 μmol·L-1 Baohuoside Ⅰ groups were 1.00±0.00,0.88±0.05,0.84±0.08,0.70±0.16 and 0.57±0.14;the relative expression levels of FoxP3 protein were 1.00±0.00,0.89±0.10,0.76±0.19,0.70±0.19 and 0.37±0.12.The differences of above indexes between the 5,10 and 20 μmol·L-1 Baohuoside Ⅰ groups and the 0 μmol·L-1 Baohuoside Ⅰ group were statistically significant(P＜0.01,P＜0.001).Furthermore,Baohuoside Ⅰ decreased the stability of FoxP3 which can be observed in the CHX pulse-chase assay at the 6 hour time point where the expression levels of FoxP3 in the control group at 0.83±0.06 compared to the experimental group at 0.55±0.11(P＜0.001).Conclusion Baohuoside Ⅰcan destabilize FoxP3 in Tregs which has been shown to reduce the immunosuppressive functions of Tregs.

Aquilaria sinensis can generate agarwood, which is closely related with endophyte. Up to now, studies mainly focused on the effects of endophytic fungi on agarwood formation, but studies about endophytic bacteria are rarely reported. In our research, the T-RFs and Shannon index of endophytic bacteria in samples of agarwood increase. The number of distinctive T-RFs fragments of corresponding samples in the same group accounted for more than 60% the number of total T-RFs fragments. In samples of no-agarwood, the dominant bacterial population are Anoxybacillus, Clostridium, Candidatus endobugula, Lysinibacillus. In samples of agarwood, the dominant bacterial population are Clostridium, Lysinibacillus, Luteimonas, phytoplasma. Besides, there are. specific T-RFs fragment in samples of agarwood and no-agarwood respectively. When we perform cluster analysis, we found samples of agarwood highly gather together and samples of no-agarwood highly gather together. This means community of endophytic bacteria emerge significant and regular changes during agarwood formation, which may be result of agarwood production, or maybe it is important reason of agarwood production. In this paper, we obtain more comprehensive and accurate community of endophytic bacteria in Aquilaria sinensis and it's variation during agarwood formation using T-RFLP, which is first study of effects of endophytic bacteria on agarwood formation, and will help to exploit resource of endophytic bacteria more reasonably.
Bacteria ; classification ; genetics ; isolation & purification ; Biodiversity ; Endophytes ; classification ; genetics ; isolation & purification ; Thymelaeaceae ; microbiology ; Wood ; microbiology

OBJECTIVETo observe the changes of intestinal flora in diarrhea type irritable bowel syndrome with Pi-wei dampness-heat syndrome (IBS-PDS).

METHODSThe seven kinds of common intestinal bacteria in feces, including enteri bacillus, enterococci, saccharomycete, bifid bacteria, lactobacillus, bacteroides and peptococcus were studied in 21 patients suffered from IBS-PDS, and compared with those in 22 patients with IBS with deficiency of Pi syndrome (DPS) and 25 healthy subjects as control.

RESULTSAs compared with the healthy subjects, the levels of enteri bacillus and enterococci were significantly increased (P<0.01), the levels of bifid bacteria, Lactobacillus and Peptococcus were significantly decreased (P < 0.01), and saccharomycete and Bacteroides were insignificantly different in patients with PDS. As compared with patients with DPS, the levels of enteri bacillus, enterococci, bifid bacteria, Lactobacillus, Peptococcus and Bacteroidaceae were significantly increased except the level of saccharomycete.

CONCLUSIONThere may be alteration of intestinal flora in patients with IBS-PDS.

Adult ; Bifidobacterium ; isolation & purification ; Diagnosis, Differential ; Diarrhea ; etiology ; microbiology ; Enterobacteriaceae ; isolation & purification ; Female ; Humans ; Intestines ; microbiology ; Irritable Bowel Syndrome ; complications ; microbiology ; Lactobacillus ; isolation & purification ; Male ; Medicine, Chinese Traditional ; Middle Aged