ABSTRACT
Background Light-induced retinal damage results in the damage of retinl pigment epithelial (RPE) cells and therefore affects the pathogenesis and development of age-related macular degeneration (AMD).Studies showed that tissue factor (TF) is overexpressed in oxidative damaged RPE cells and the choroidal neovascularization (CNV) of AMD,speculating that the suppression of TF can prevent the damage of RPE cells and inhibit CNV.Objective This study was conducted to observe the protective effects of TF targeting peptide (TFTP),a new drug of autologous synthesis,on human RPE-cells induced by blue light.Methods Human RPE cells were isolated from donor eye and cultured.Cultured cells were divided into blank control group,model group and TFTP treated group.Light-induced RPE cell damage model was established by exposuring the cells in the blue light of (4.0±-0.5) mW/cm2 for 12 hours in the model group,and different concentrations (10,100,150,200,300 μmol/L) of TF-TP were added into the medium to pretreat the cells for 24 hours and then exposed the cells to the blue light for 12 hours in the TF-TP groups.The cell viability was determined by CCK-8 assay.The morphology and ultrastructure in the cells were observed under the inverted microscope and transmission electron microscope.The apoptosis of the cells was assayed by Hoechst staining.The expressions of TF and apoptosis-related protein bax,bcl-2 in the cells were determined by Western blot.Results CCK-8 assay showed that there was no significant difference in the cell viability among blank control group and different concentrations TF-TP groups (F=2.15,P =0.11).The cell survival rate of blank control group,model group and 150 μmol/L TF-TP group was (100.0±0.00) ％,(43.79±6.55) ％ and (63.45±3.57) ％,and the survial rate was increased in the 150 μmol/L TF-TP group compared with the model group (P =0.00),and 150 μmol/L was detemined as a optimal concentration of TF-TP.A lot of shrinkage,deformation,suspension cells were exhibited under the optical microscope,and decrease of microvilli structure,rupture of mitochondrial cristae and vacuolar degeneration of the cells were found in the model group,and the damage of the cells were evidently lightened in the 150 μ mol/L TF-TP group.The apoptosis rate of the cells were (0.98 ±0.19)％,(9.98 ±0.82) ％ and (5.73 ±0.88) ％ in the blank group,model group and 150 μmol/L TF-TP group,respectively,with a significant difference among the groups (F =206.18,P =0.00),and the apoptosis rate of the cells in the 150 μmol/L TF-TP group was significantly lower than that in the model group (P＜0.05).Compared with the blank control group,the relative expression of bax and TF was obviously increased and that of bcl-2 was decreased in the model group;while the expression of bax and TF was lower,and that of bcl-2 was higher in the 150 μmol/L TF-TP group compared with the model group (all at P ＜ 0.05).Conclusions Pretreation of TF-TP can lessen cell apoptosis and increase cell survival rate and therefore plays a protective role to blue light-induced human RPE cells possibly by inhibiting bax/bcl-2 apoptotic pathways mediated by TF.
ABSTRACT
<p><b>OBJECTIVE</b>To compare fecal microbiota and metabolites between colorectal cancer (CRC) patients and healthy population.</p><p><b>METHODS</b>Feces from fifteen CRC patients and twelve normal people were analyzed by using pyrosequencing and gas chromatography mass spectrometry(GC/MS).</p><p><b>RESULTS</b>There were no significant differences in the overall microbial community structure associated with the disease state, but 18 bacterial genera were underrepresented or overrepresented in the CRC samples. GC-MS profiling revealed higher concentrations for 9 kinds of amino acids and metabolites of short-chain fatty acids, lower concentrations for 3 kinds of unsaturated fatty acids and 2 kinds of glycerin and ursodeoxycholic acid in stool samples from CRC patients. Correlative analysis between the combined datasets revealed some potential relationships between stool metabolites and certain bacterial species.</p><p><b>CONCLUSIONS</b>There are significant differences in fecal metabolites and the relative abundance of certain types of bacteria between CRC patients and healthy people, which can provide insight into microbial functions occurring in a cancer environment and will help direct future mechanism studies.</p>
ABSTRACT
AIM: To investigate the expression of objective gene and the immunosuppressive effect of naked DNA vaccine pγ1.Ig H.SPTT targeting tissue factor by intrasplenic inoculation on colorectal cancer ( CRC) .METH-ODS:A special naked DNA vaccine which carried the SPTT peptides and immunoglobulin H chain gene ( named pγ1.Ig H.SPTT DNA plasmid ) was constructed by molecular biological techniques .After a single injection of this plasmid into the spleen, the concentrations of transgene product SPTT-Ig H in the peripheral blood at different time points were detected by ELISA, and the plasmid transfection efficiency and characteristics were analysis by PCR and Southern blotting .The immu-nologic effect of the plasmid on the CRC was observed in the mice .RESULTS:The strongest expression of SPTT-Ig H was observed during the 4th week after a single injection of the plasmid , which began to decline at the 12th week and disap-peared at the 16th week.The concentration of SPTT-Ig H in the peripheral blood at the 2nd week after transfection of plas-mid was 7.2 μg/L, then increased gradually , and reached a peak of 13.11μg/L at the 8th week.The plasmid-transcrip-tional gene was only expressed in the spleen , and was not detected in the lymph nodes , bone marrow, liver, kidney, and other organisms.Transfection of pγ1.Ig H.SPTT into the spleen had inhibitory effects on colorectal cancer as compared with control group .CONCLUSION:Naked DNA vaccine pγ1.Ig H.SPTT stimulates the immune response for protecting the body against colorectal cancer , which is a safe and effective method for CRC immunotherapy .