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Background Epithelial-myofibroblast transition (EMT) of human lens epithelial cells (LECs) induced by transforming growth factor-β2 (TGF-β2) is the main mechanism in the pathogenesis of posterior capsular opacification(PCO).Seeking an effective drug capable of inhibiting this process is important for the prevention and treatment of PCO.Objective The purpose of this study was to investigate the inhibitory effect of rapamycin (RAPA)on the proliferation of human LECs and TGF-β2-induced EMT.Methods Human LEC strain(SRA01/04)was cultured in DMEM with high glucose and 10％ fetal bovine serum.The cells were consequently cultured in serumfree DMEM with 5 mg/L TGF-β2,TGF-β2+10 mg/L RAPA,TGF-β2 + 100 mg/L RAPA,TGF-β2 + 1000 mg/L RAPA or TGF-β2 +10 000 mg/L RAPA for 72 hours,and SRA01/04 cultured in serum-free DMEM were used as control.The proliferation rate(A490)of SRA01/04 in the different groups was detected using the MTT assay and the rate of inhibition of RAPA was calculated.The expressions of the α-smooth muscle actin(α-SMA) and E-cadherin(E-cad)mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot,respectively.The changes in the expression of α-SMA and E-cad in SRA01/04 were evaluated by Western blot 24,48 and 72 hours after TGF-β2 +400 mg/L RAPA treatment.Results The A490 value of SRA01/04 was 0.680±0.020,0.550±0.013,0.480±0.014,0.400±0.011 and 0.200±0.019 in the control group,TGF-β2 group,TGF-β2 + 10 mg/LRAPA group,TGF-β2 + 100 mg/L RAPA group,TGF-β2 + 1000 mg/L RAPA and TGF-β2 + 10 000 mg/L RAPA group,respectively,showing a gradually declining trend in SRA01/04 rate of proliferation with increasing RAPA concentrations (F =101.920,P =0.000).RT-PCR and Western blot assay showed that the relative expression levels of α-SMA mRNA (α-SMA mRNA/β-actin mRNA)and protein (α-SMA/β-actin)in the cells were significantly increased in the TGF-β2 treatment group.However,with exposure to RAPA,the relative expression levels of α-SMA mRNA and protein were significantly lowered with increasing RAPA concentrations,but the expression levels of E-cad mRNA and protein were raised (α-SMA mRNA:F =294.660,P =0.000 ; α-SMA protein:F =346.950,P =0.000 ; E-cad mRNA:F =264.250,P =0.000 ; E-cad protein:F =317.327,P =0.000).In addition,after exposure to 400 mg/L RAPA,the expression levels of α-SMA protein gradually reduced and those of E-cad protein gradually increased with increasing treatment durations,showing significant differences among the different time points (α-SMA:F =693.864,P =0.000 ;E-cad:F=369.286,P =0.000).Conclusions RAPA can inhibit the proliferation of SRA01/04 in vitro and arrest EMT of SRA01/04 induced by TGF-β2 in a dose-and time-dependent manner.
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Background Ultraviolet irradiation promotes cellular apoptosis by affecting the mitochondrial transmembrane potential,including human lens epithelial cells (LECs).Gene associated with retinoid-interferoninduced mortality-19 (GRIM-19),a cell death regulatory protein,is essential for the assembly and function of mitochondrial complex Ⅰ.However,whether LECs apoptosis induced by ultraviolet irradiation is related to GRIM-19 is still unclear.Objective The purpose of this study was to investigate the relationship between the apoptosis of human LECs caused by ultraviolet with GRIM-19 expression in vitro.Methods Human LEC line(SRA01/04)was cultured in α-MEM containing 10％ fetal bovine serum.The cells were exposed to ultraviolet ray at doses of 0,30,60,90,120 or 150 mJ/cm2 when cell growth reached the logarithmic phase and 80％ confluency.The rate of apoptosis of the cells was assayed using flow cytometry,and the level of expression and relative amount of GRIM-19 protein (GRIM-19/β-actin) were detected by Western blot.The relationship between apoptosis and the GRIM-19/β-actin value among the different treatment groups was compared using One-way ANOVA,and the correlation of LECs apoptosis rate and GRIM-19 expression level was assessed by Pearson linear analysis.Results A significant difference was found in the apoptosis rate among the different treatment groups(F=149.32,P＜0.01).Compared with the 0 mJ/cm2 ultraviolet irradiation group,the apoptosis rate of LECs was significantly increased in the 60,90,120 and 150 mJ/cm2 ultraviolet irradiation groups (q =17.02,-25.20,-29.41,-8.61,P ＜ 0.01).The expression of the GRIM-19 protein in the LECs suspension was enhanced by ultraviolet irradiation at 60,90,120 and 150 mJ/cm2.The relative expression of the GRIM-19 protein (GRIM-19/β-actin) was significantly different among the various groups (F=6.87,P＜0.05),and the GRIM-19/β-actin values in the 60,90,120,150 mJ/cm2 ultraviolet irradiation groups were elevated in comparison with the un-irradiated group(2.01±0.76,2.98± 1.80,3.97± 1.61,2.42± 1.28 vs.0.56±0.23),which showed statistically significant differences (q =4.12,-5.04,-7.09,-3.85,P ＜ 0.01).In addition,a positive correlation was seen between the rate of apoptosis and the expression of the GRIM-19 protein(r=0.71,P＜0.01).Conclusions GRIM-19 is expressed in normal human LECs.The apoptosis of human LECs accompanies the up-regulation of GRIM-19.The expression of GRIM-19 in LECs increases with ultraviolet irradiation in a doseindependent manner.
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Objective To investigate the relationship between drinking-water type of endemic arsenicosis and adult carotid artery atherosclerosis. Methods In 2009, 285 participants aged over 40 from drinking-water type of endemic arsenism areas and 293 residents aged over 40 from control areas were investigated in Yingxian county,Shanxi province. Portable-type B mode color ultrasound was used to examine the carotid artery of all participants.The carotid atherosclerosis were diagnosed and graded through the ultrasonograms. Content of water arsenic and hair arsenic of 10 people randomly selected in every villages were detected. Results A total of 5 villages with drinkingwater type of endemic arsenicosis as observation group and 5 villages without drinking-water type of endemic arsenicosis as control group were investigated. The prevalence rates of adult carotid atherosclerosis within observation group were 35.09%(20/57), 55.74%(34/61), 38.46%(20/52), 36.51%(23/63) and 46.15%(24/52), respectively,and standardized prevalence rates were 32.5%, 33.8%, 34.9%, 46.2% and 47.3%, respectively and the prevalence rates of adult carotid atherosclerosis within control group were 18.18%(10/55), 30.77%(16/52), 20.00%(10/50),18.67% (14/75) and 21.31% ( 13/61 ), respectively; the standardize prevalence rates were 22.4%, 17.7%, 10.7%,24.6%, 18.9%, respectively. The standardize prevalence rates were higher in observation group [39.50%(113/285) ]than that in control group[39.50%(113/285), T = 26, P ＜ 0.01 ]. The severity of adult carotid atherosclerosis (composition of 4 - 7 scores ) was compared between observation group [ 17.70%(20/113 )] and control group [ 14.06% (9/64) ], and the difference was insignificant(x2 = 0.26, P ＞ 0.05). Conclusions The prevalence rate of carotid atherosclerosis in drinking-water type of endemic arsenicosis areas is higher than that of the control areas.The study provides evidence that arsenic poisoning can cause atherosclerosis.
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Objective To investigate the disease situation of adult carotid atherosclerosis in water-related endemic fluorosis areas in Heilongjiang province in 2008 so as to explore the relationship between water-related endemic fluorosis and incidence of carotid atherosclerosis disease. Methods A total of 266 participants over the age of 40 from four villages in Zhaozhou county with water fluoride ≥ 1.0 mg/L in a fluorosis area and 283 residents over the age of 40 from four villages in Tailai county with water fluoride ＜ 1.0 mg/L in a nonfluorosis area were investigated. A portable-type B mode color ultrasound was used to examine the left carotid artery of all participants.The carotid atherosclerosis was diagnosed and graded through the ultrasonograms. Results The prevalence rates of carotid atherosclerosis in the 4 fluorosis villages(Xinfeng, Taipingshan, Baochan and Houzheng villages) were 47.3% (35/74), 63.5%(40/63), 73.3%(33/45) and 60.7%(51/84), respectively, and in the 4 nonfluorosis villages(Hala,Qianxing, Sanjia and Ailin villages) were 32.7% (17/52), 32.9% (24/73), 39.2% (31/79) and 30.4% (24/79),respectively. The prevalence rates of carotid atherosclerosis in every villages standardized by age were 47.3% ,63.5%,73.3% ,60.7% and 34.7% ,36.3% ,43.0% ,41.3%, respectively. Statistic method used was Wilcoxon two sample test and the differences were significant(T = 26, P ＜ 0.05 ). Total carotid atherosclerosis positive rate standardized by age in the 4 fluorosis villages was significantly higher than that in the 4 nonfluorosis villages[57.5%(153/266) ,37.8% (107/283), x2 = 21.36, P ＜ 0.01 ]. After standardized by age, the severity of carotid atherosclerosis was significantly different between fluorosis villages and nonfluorosis villages (x2 = 36.15, P ＜ 0.01 ). Conclusion The prevalence rate of carotid atherosclerosis in endemic fluorosis area is higher than that in nonfluorosis area.
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Objective To study the practicality and repeatability of B-mode ultrasonography on diagnosis and score of carotid atherosclerosis. Methods Ninety-nine B-mode ultrasonography pictures of carotid atherosclerosis were obtained from field investigation. According to the established standard(normal scored as 0, other scored as 1 - 7 by the severity), the 99 pictures were diagnosed and scored once together by three trained inexperienced members of the research team. Ten days later, these pictures were diagnosed and scored respectively, by the three members.The diagnostic results were analyzed with statistical methods to test the feasibility and repeatability of this set of diagnostic and scoring system. Results The joint diagnostic results by the three group members were regard as standard with scores 0 in 27 cases, 1 in 27 cases, 2 in 10 cases, 3 in 4 cases, 4 in 13 cases, 5 in 8 cases, 6 in 6 cases, and 7 in 4 cases. Independent diagnostic results by member A was 0 in 29 cases, 1 in 24 cases, 2 in 11 cases, 3 in 2 cases, 4 in 12 cases, 5 in 11 cases, 6 in 6 cases, and 7 in 4 cases. Independent diagnostic results by member B was 0 in 29 cases, 1 in 22 cases, 2 in 14 cases, 3 in 4 cases, 4 in 13 cases, 5 in 5 cases, 6 in 8 cases, and 7 in 4 cases. Independent diagnostic results by member C was 0 in 28 cases, 1 in 25 cases, 2 in 8 cases, 3 in 5 cases, 4 in 14 cases, 5 in 10 cases, 6 in 4 cases, and 7 in 5 cases. Comparison of the 4 diagnostic scores, the difference was not statistically significant(F = 0.019, P ＞ 0.05). Joint diagnostic results of the three members were compared with the standard and correlation coefficient were 0.977,0.987,0.932, respectively(all P ＜ 0.01 ). The correlation coefficient of diagnostic results of each member were 0.969,0.935,0.928, respectively (all P ＜0.01 ). Diagnostic results of each member were consistent with the standard and the compliance rate were 88.9% (88/99) ,90.9%(90/99) and 86.9%(86/99), respectively. Conclusions B-mode ultrasonography is a non-injure method for diagnosis of carotid atherosclerosis in epidemiology investigation. The method is easy to grasp and has a good repeatability.
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Objective To assess the prevalence of carotid atherosclerosis in adult patients with KaschinBeck disease(KBD) in endemic area, and to investigate the association between KBD and atherosclerosis. Methods B-mode ultrasound was used to investigate carotid atherosclerosis in patients with KBD over 40 years old in Shanhe farm and Shangzhi city and residents in Shuangcheng county in 2009. Ultrasonography images were saved and graded according to intima media morphology. Results The prevalence rates of carotid atherosclerosis in patients with KBD from the 5 villages affiliated to Shanhe farm were 50.0%(26/52),37.3%( 19/51 ),42.5%(30/71 ),58.2% (39/67) and 44.0% (22/50) , respectively. The prevalence rates of carotid atherosclerosis in patients with KBD from the 5 villages affiliated to Shangzhi city were 63.0% (34/54), 45.3% (24/53), 47.2% (25/53), 60.0% (30/50)and 51.9% (28/54), respectively. The prevalence rates of carotid atherosclerosis in people without KBD in the 5 control villages affiliated to Shuangcheng county were 63.9% (46/72),43.1% (31/72),57.1% (40/70),46.9% (30/64) and 53.0%(35/66), respectively. Ten villages in KBD endemic area and 5 villages in control area were investigated. The prevalence rates of carotid atherosclerosis in the 15 villages were standardized and compared by Wilcoxon test. The differences were not significant between two KBD endemic areas and one control( T = 25.0,24.5,all P ＞ 0.05). There was no significant difference in prevalence of atherosclerosis in the 3 areas(x2 = 0.36, P＞0.05). Comparing the severity of carotid atherosclerosis in 3 locations, the difference was statistically significant (x2 = 15.28, P ＜ 0.05) and there was significant difference statistically between the subjects in Shangzhi city and that in the control group(x2 = 8.92, P ＜ 0.05). There was no significant difference between the detection rate of atherosclerosis and the severity of KBD, and similar results in the grade of atherosclerosis and the severity of KBD were found (x2 = 1.88,5.07, all P ＞ 0.05). Conclusions The prevalence rate of carotid atherosclerosis in historical endemic area in Heilongjiang province is not higher than that in the control area.
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Objective To survey the prevalence rates of adult carotid atherosclerosis in water-sourceoriginated high iodine area in Shandong province in 2008 and discuss the relationship between water-sourceoriginated high iodine and carotid atherosclerosis occurrence. Methods Yuncheng county was chosen as observation area which included Dachenlou, Weigai, Liuyi and Ershilipu villages. Juye county was chosen as control that included Zhangxi, Bihainan, Xiaguan and Qiji villages. Two hundred and ninety-nine participants aged over 40 from water-source-originated high iodine areas and 323 residents aged over 40 from normal iodine areas were investigated.Portable-type B mode color Doppler was performed to examine the carotid artery of all participants. The adult carotid atherosclerosis was diagnosed and graded through the ultrasonograms. Results The prevalence rates of adult carotid atherosclerosis in the 4 water-source-originated high iodine villages of Dachenlou, Weigai, Liuyi and Ershilipu were 47.1% (33/70) ,62.2% (51/82) ,67.5% (52/77) and 58.6% (41/70), respectively and the prevalence rates of adult carotid atherosclerosis in the 4 normal iodine villages of Zhangxi, Bihainan, Xiaguan and Qiji were 40.7%(35/86),40.8% (31/76),38.2% (34/89) and 37.5% (27/72), respectively. The prevalence rates of adult carotid atherosclerosis standardized by age in the 4 high iodine villages and the 4 normal iodine villages were 45.81% ,58.18% ,61.63%,55.34% and 34.66%,36.25%,43.01% ,41.30%, respectively. The prevalence rates were higher in high iodine villages than that in control villages(T = 26, P ＜ 0.05 ). There were 136 people scored "0", 120 people scored "1 -3",43 people scored "4 - 7", and total 299 people. There were 201 people scored "0", 87 people scored "1 - 3", 35 people scored "4 - 7", and total 323 people. The expected prevalence rates and severity of adult carotid atherosclerosis was significantly different between high iodine area and normal iodine area (x2 = 17.54,18.42, all P ＜ 0.01).Conclusion The prevalence rate and severity of adult atherosclerosis in water-source-originatod high iodine area is higher than that in normal iodine area.