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Objectives To investigate the expressions of miR-101 and COX-2 in colorectal cancer. Methods Thirty-twocolorectal cancer specimens and paired paracancer tissues were collected for assessment of miR-101 and COX-2 expressions by real-time quantitative PCR. Thecorrelation between miR-101 and COX-2 , as well as their correlations with the pathologywere analyzed. Results The expression level of miR-101 and COX-2 mRNA in the cancer tissues was significant difference between the cancer and para-cancer tissues (P 0.05). Conclusion Down-regulated miR-101 expression andparallel COX-2 overexpression hasbeen linked to colorectal cancer. miR-101 and COX-2might be thepotentialdiagnostic markers and therapeutic targets.
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Objective To investigate the influence of baicalin in human colon cancer SW480 cells,and to clarify its mechanism.Methods The SW480 cells were cultured and divided into blank control and 25,50 and 100 μmol·L-1 baicalin groups.The proliferation activity was detected with CCK-8 assay.The morphological changes of SW480 cells were detected by Annexin Ⅴ-FITC and DAPI coloration.The protein expression levels of Bcl-2,caspase 3 and caspase 9 were detected by Western blotting method. Results The CCK-8 assay results showed that the proliferation activities of SW480 cells in 25,50 and 100 μmol· L-1 baicalin groups were decreased significantly compared with blank control group at the time points of 24 h,48 h and 72 h (P <0.01),the proliferation activities of SW480 cells in 25 μmol·L-1 baicalin groups were decreased significantly compared with blank control group at the time points of 48 and 72 h (P <0.01).Cell shrinkage and nucleus fragmentation were observed in the SW480 cells after treated with 50 μmol·L-1 baicalin for 48 h.The Western blotting assay results showed that compared with blank control group,the protein expression levels of caspase 3 and caspase 9 in 25,50 and 100 μmol· L-1 baicalin groups were increased significantly (P <0.05 or P <0.01),and the protein expression levels of Bcl 2 in 25, 50 and 100 μmol·L-1 baicalin groups were decreased significantly (P <0.05 or P <0.01).Conclusion Baicalin can induce the apoptosis in SW480 cells,and the effect might be involved with the mitochondrial apoptotic pathway.
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Objective To construct and identify a novel IL-10 delivery system by transforming a hIL-10-containing plasmid into B.longum (BL -hIL -10).Methods A plasmid vector pBADs -GFP was selected which had been built by previous test and biosynthetic hIL-10 plasmid,through double enzyme digestion and enzyme reaction,to construct and identify PBADs-hIL-10 shuttle plasmid,then to synthesis BL-hIL-10.hIL-10 was expressed and secreted into the culture supernatant of BL-hIL-10 after 0.2% L-arabinose induction in vitro as examined by Western blot,enzyme-linked immunosorbent assay (ELISA)and RT-PCR;Culture supernatants and bacterium pellets were collected after continuous culture for 12,24 and 36h,respectively.hIL-10 was expressed and secreted into the culture supernatant of BL-hIL-10 after 0.2% L-arabinose induction in vitro as examined by Western blot,enzyme -linked immunosorbent assay (ELISA)and RT -PCR;Culture supernatants and bacterium pellets were collected after continuous culture for 12,24 and 36h,respectively.Results The BL-hIL-10 bacterial strain that can stably express hIL-10 factor was successfully screened out,and the levels of hIL-10 in both superna-tant and cell pellet were similarly reached maximum at 24h of culture.Conclusion BL-hIL-10 as a novel oral hIL-10 delivery system has been successfully established,which established a basis for the treatment of IBS with transgenic Bifidobacterium.
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<p><b>OBJECTIVE</b>To observe the effect of TSP-2, the antibody of Toll-like receptor 2 extracellular domain, on the expression and DNA-binding activity of nuclear factor-κB (NF-κB) p65 protein in mice with ulcerative colitis (UC).</p><p><b>METHODS</b>Sixty BALB/c mice were randomized equally into normal control group, UC model group, TSP-2 treatment group, and rabbit IgG treatment group. In the latter 3 groups, the mice were fed with 5% DSS (C6H7Na3O14S3) solution for 7 days to induced UC, followed then by treatment with daily injections of TSP-2 or rabbit IgG as appropriate for 7 days. The disease activity index was recorded during the treatment. The colitis tissues were collected after the treatments for HE staining and detecting the expression and DNA-binding activity of NF-κB p65 in the colon mucosa by Western blotting and ELISA.</p><p><b>RESULTS</b>The DNA binding activity and expressions of NF-κB P65 protein increased significantly in UC model group (P<0.05). TSP-2 treatment group significantly decreased the disease activity index (P<0.05) and lowered the DNA-binding activity and expression of NF-κB P65 protein (P<0.05) in the UC mouse models, while rabbit IgG produced no such effects (P>0.05).</p><p><b>CONCLUSION</b>TSP-2 can suppress the DNA-binding activity and protein expressions of NF-κB P65 and regulate excessive immune response in the intestines to ameliorate ulcerative colitis in mice.</p>