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Objective:To investigate the inhibitory effect of CLC-2 chloride channel targeted blocking on fibrosis of human conjunctival fibroblasts (HConF).Methods:HConF were divided into blank control group, lipofectamine 2000 (Lipo2000) group, nonsense small interfering RNA (siRNA) group, and CLC-2 siRNA transfected group.The HConF were cultured in medium containing the corresponding transfection reagents according to grouping.No intervention was given to blank control group.The expression level of CLC-2 mRNA of HConF was detected by real-time fluorescence quantitative PCR; absorbance ( A) value indicating the proliferative ability of HConF was determined by CCK-8 kit; the apoptosis ratio of HConF was tested by flow cytometry; the migration ability of HConF was identified by cell scratch test and Transwell migration assay; the contraction rate of HConF was assayed by collagen contraction test; the expression levels of collagenⅠ, collagen Ⅲ, PI3K, Akt, p-PI3K and p-Akt proteins were measured by Western blot. Results:Significant differences were found in relative expression levels of CLC-2 mRNA and A value among four groups ( F=90.110, 198.680; both at P<0.001). The relative expression level of CLC-2 mRNA and A value were significantly lower in CLC-2 siRNA transfected group than nonsense siRNA group, showing statistically significant differences (both at P<0.001). The proportion of apoptotic HConF in blank control group, Lipo2000 group, nonsense siRNA group, and CLC-2 siRNA transfected group was (4.78±1.10)%, (4.54±1.51)%, (4.82±0.88)% and (28.90±0.91)%, respectively, and a statistically significant difference was found ( F=363.260, P<0.001). The proportion of apoptotic HConF was significantly higher in CLC-2 siRNA transfected group than nonsense siRNA group, with a statistically significant difference ( P<0.001). Statistically significant differences were found in cell migration rate and the number of migrating cells among four groups ( F=74.493, 1 625.431; both at P<0.01). The cell migration rate of HConF in CLC-2 siRNA transfected group was significantly lower and the number of migrating cells was significantly smaller than those of nonsense siRNA group, with statistically significant differences (both at P<0.001). A statistically significant difference in contraction rate was found among four groups ( F=104.692, P<0.001). The contraction rate of HConF was significantly lower in CLC-2 siRNA transfected group than nonsense siRNA group, and the difference was statistically significant ( P<0.001). Statistically significant differences were found in relative expression levels of collagen Ⅰ and collagen Ⅲ proteins, p-PI3K/PI3K ratio, and p-Akt/Akt ratio among four groups ( F=112.073, 456.931, 340.889, 43.021; all at P<0.001). The relative expression levels of collagen Ⅰ and collagen Ⅲ proteins, p-PI3K/PI3K ratio and p-Akt/Akt ratio in CLC-2 siRNA transfected group were significantly lower than those of nonsense siRNA group, showing statistically significant differences (all at P<0.05). Conclusions:Targeted blocking of CLC-2 chloride channel gene expression can inhibit fibrosis of HConF by promoting apoptosis of HConF through PI3K/Akt signaling pathway and inhibit fibrotic processes such as cell migration, collagen synthesis and collagen contraction.
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Purpose@#Sedentary behavior attributes to the increased risk of some cancers and all-cause mortality. The evidence is limited for the association between television (TV) viewing time, a major sedentary behavior, and risk of colorectal cancer death. We aimed to examine this association in Japanese population. @*Materials and Methods@#A prospective cohort study encompassed of 90,834 men and women aged 40-79 years with no prior history of colorectal cancer who completed a self-administered food frequency questionnaire, and provided their TV viewing information. The participants were followed-up from 1988-1990 to the end of 2009. The hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated by the Cox proportional hazard regression for risk of colorectal cancer mortality according to TV viewing time. @*Results@#During the median 19.1-year follow-up period, we documented 749 (385 men and 364 women) colorectal cancer deaths. The multivariable-adjusted HRs for mortality from colorectal cancer were 1.11 (0.88-1.41) for 1.5 to < 3 hr/day, 1.14 (0.91-1.42) for 3 to < 4.5 hr/day and 1.33 (1.02-1.73) for ≥ 4.5 hr/day in comparison to < 1.5 hr/day TV watching; p-trend=0.038, and that for 1-hour increment in TV viewing time was 1.06 (1.01-1.11). Moreover, the multivariable-adjusted HR (95%CI) of colon cancer for 1-hour increment in TV viewing time was 1.07 (1.02-1.13). Age, body mass index, and level of leisure-physical activity did not show significant effect modifications on the observed associations. @*Conclusion@#TV viewing time is associated with the increased risk of colorectal cancer mortality among Japanese population, more specifically colon rather than rectal cancer.
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Purpose@#Sedentary behavior attributes to the increased risk of some cancers and all-cause mortality. The evidence is limited for the association between television (TV) viewing time, a major sedentary behavior, and risk of colorectal cancer death. We aimed to examine this association in Japanese population. @*Materials and Methods@#A prospective cohort study encompassed of 90,834 men and women aged 40-79 years with no prior history of colorectal cancer who completed a self-administered food frequency questionnaire, and provided their TV viewing information. The participants were followed-up from 1988-1990 to the end of 2009. The hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated by the Cox proportional hazard regression for risk of colorectal cancer mortality according to TV viewing time. @*Results@#During the median 19.1-year follow-up period, we documented 749 (385 men and 364 women) colorectal cancer deaths. The multivariable-adjusted HRs for mortality from colorectal cancer were 1.11 (0.88-1.41) for 1.5 to < 3 hr/day, 1.14 (0.91-1.42) for 3 to < 4.5 hr/day and 1.33 (1.02-1.73) for ≥ 4.5 hr/day in comparison to < 1.5 hr/day TV watching; p-trend=0.038, and that for 1-hour increment in TV viewing time was 1.06 (1.01-1.11). Moreover, the multivariable-adjusted HR (95%CI) of colon cancer for 1-hour increment in TV viewing time was 1.07 (1.02-1.13). Age, body mass index, and level of leisure-physical activity did not show significant effect modifications on the observed associations. @*Conclusion@#TV viewing time is associated with the increased risk of colorectal cancer mortality among Japanese population, more specifically colon rather than rectal cancer.
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BACKGROUND@#This study aimed to describe the status of alcohol consumption and drug use among young adults as well as their determinants.@*METHODS@#We conducted a cross-sectional study of 356 young adults (aged 18 to 24 years) living in Palau in 2013. The prevalence of self-reported alcohol and marijuana usage were compared within and between sexes, age groups, ethnicities, and education levels.@*RESULTS@#The proportion of current drinking was higher in people aged 21-24 than in those aged 18-20 (73.2% vs. 60.9%, p = 0.09 in men and 48.3% vs. 30.0%, p = 0.02 in women), while that of marijuana use did not differ between the age groups. The proportions of current drinking and marijuana use were higher in Palauan than in other ethnicities (current drinking: 70.6% vs. 40.6%, p = 0.005 in men and 38.8% vs. 16.6%, p = 0.04 in women; lifetime marijuana use: 80.0% vs. 52.9%, p = 0.02 in men and 56.1% vs. 30.6%, p = 0.09 in women). The proportion of frequent (3 times or more) marijuana users was higher for the lower educated than for the higher educated (62.5% vs. 32.1%, p < 0.001 in men and 33.9% vs. 24.4%, p = 0.12 in women).@*CONCLUSIONS@#Sex, age, ethnicity, and education were significant determinants of alcohol and marijuana use.
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Female , Humans , Male , Young Adult , Age Factors , Alcohol Drinking/ethnology , Cross-Sectional Studies , Educational Status , Marijuana Use/ethnology , Palau/epidemiology , Prevalence , Sex FactorsABSTRACT
Objective To study the effect and mechanisms of chloride channel blocker 5-Nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) on thansforming growth factor β1 (TGF-β1) induced human conjunctival fibroblasts (HConF) fibrosis.Methods Cell counting kit (CCK-8) was used to screen out the optimal TGF-β1 treatment time and the optimal NPPB concentration.The cells were divided into control group,TGF-β1 treatment group and TGF-β1+NPPB group.Cell proliferation and cell cycle were detected by CCK-8 and flow cytometer,respectively.Cell migration ability were observed by scratch and transwell migration assays.Western blot and Real time-PCR were used to detect the expression of collagen Ⅰ (COL-Ⅰ),fibronectin (FN) and α-smooth muscle actin (α-SMA).The phosphorylation level of PI3K and Akt were measured by Western blot.Results TGF-β1 promotes cell proliferation in a time-dependent manner.There was no statistically significant difference in A values between 48 hours and 72 hours after TGF-β1 treatment (P =0.064).Forty-eight hours was selected as the most appropriate time for TGF-β1 treatment.NPPB inhibited HConF cell proliferation in a concentration-dependent manner.Compared with the control group,the proliferation A values of cells in the 50 mol/L and 100 mol/L NPPB groups were significantly reduced (P =0.020,0.000),and 100 mol/L was selected as the optimal concentration of NPPB.The cell proliferation A value,migration area and migration cell number of TGF-β1 +NPPB group were significantly lower than those of TGF-β1 treatment group (all at P＜0.05).Compared with the control group and TGF-β1 +NPPB group,the proportion of G1 phase cells in the TGF-β1 treatment group was reduced,and the proportion of cells in the S phase and G2/M phase were increased,with statistically significant differences between them (all at P ＜ 0.05).The protein and mRNA expression of α-SMA,COL-Ⅰ and FN in the TGF-β1 treatment group were higher than those in the control group and TGF-β1+NPPB group,with statistically significant differences between them(all at P＜0.05);the ratios of p-PI3K/PI3K and p-Akt/Akt in the TGF-β1 treatment group were significantly higher than those in the control group and TGF-β1 +NPPB group,with statistically significant differences between them (all at P＜0.05).Conclusions NPPB may inhibit TGF-β1 induced HConF fibrosis process by inhibiting phosphorylation of PI3K and Akt.
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PURPOSE: The evidence on effects of TV viewing time among premenopausal and postmenopausal women for breast cancer risk remains controversial and limited. MATERIALS AND METHODS: A prospective study encompassing 33,276 (17,568 premenopausal, and 15,708 postmenopausal) women aged 40-79 years in whom TV viewing time, menstrual, and reproductive histories were determined by a self-administered questionnaire. The follow-up was from 1988 to 2009 and hazard ratios (HRs) with 95% confidence intervals (CIs) of breast cancer incidence were calculated for longer TV viewing time in reference to shorter TV viewing time by Cox proportional hazard models. RESULTS: During 16.8-year median follow-up, we found positive associations between TV viewing time and breast cancer incidence with a borderline significant trend among total women and a significant trend among postmenopausal women. Among total women, the multivariable HRs (95% CIs) for risk of breast cancer in reference to < 1.5 hr/day of TV viewing time were 0.89 (0.59-1.34) for 1.5 to < 3.0 hr/day, 1.19 (0.82-1.74) for 3.0 to < 4.5 hr/day, and 1.45 (0.91-2.32) for ≥ 4.5 hr/day (p for trend=0.053) and among postmenopausal women, the corresponding risk estimates were 1.10 (0.42-2.88), 2.54 (1.11-5.80), and 2.37 (0.92-6.10) (p for trend=0.009), respectively. CONCLUSION: Prolonged TV viewing time was associated with increased risk of breast cancer, especially among postmenopausal women.