Objective To investigate the protective effects of selenium on nitric oxide(NO)-mediated myocardial apoptosis.Methods The AC16 cardiomyocyte cultured in vitro were divided into control group,selenium treatmentgroup,sodium nitroprusside(SNP) treatment group and selenium + SNP treatment group,SNP was the exogenous NO donor.There was no intervention in the control group,and an equal volume of the culture solution was added to the treatment groups.The selenium treatment group added a dose of 100 μg/L of selenium,the SNP treatment group added a dose of 1.0 mmol/L of SNP,and the selenium + SNP treatment group was pretreated by 100 μg/L selenium for 4 h followed by 1.0 mmol/L SNP;the cells or supernatants were collected after 24 h of culture.The content of NO was detected by Griess method in supernatants.The level of cell reactive oxygen species was detected by flow cytometry.The changes of cell mitochondrial membrane potential and apoptosis were observed under fluorescence microscope.The real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expression levels of apoptosis-related genes B-cell lymphoma-2 (Bcl-2) associated X protein (Bax) and Bcl-2,respectively.Results The NO content in the control group,selenium treatment group,SNP treatment group and selenium + SNP treatment group were (10.3 ± 1.8),(9.2 ± 2.1),(15.2 ± 3.5),(14.3 ± 2.6) μmmol/L,respectively;SNP had a main effect on NO content (F =23.33,P ＜ 0.05).The cell reactive oxygen species were 31.63 ± 1.40,29.52 ± 2.86,60.62 ± 4.83,50.08 ± 2.41,respectively;selenium and SNP had main effects on reactive oxygen species (F =12.19,187.20,P ＜ 0.05),selenium combined with SNP had an interactive effect on reactive oxygen species (F =5.42,P ＜ 0.05).The cell mitochondrial membrane potential levels were 0.42 ± 0.11,0.37 ± 0.07,7.25 ± 1.91,and 5.21 ± 1.59,respectively;selenium and SNP had main effects on cell mitochondrial membrane potential levels (F =14.21,440.01,P ＜ 0.05),selenium combined with SNP had an interactive effect on cell mitochondrial membrane potential levels (F =12.89,P ＜ 0.05).Selenium had main effects on nuclear pyknosis ratio,Bcl-2 mRNA and Bax protein expressions (F =9.52,10.84,22.17,P ＜ 0.05);SNP had main effects on nuclear pyknosis ratio,Bax and Bcl-2 mRNA expressions,and Bcl-2 protein expression (F =192.86,21.90,16.09,18.39,P ＜ 0.05);selenium combined with SNP had an interactive effect on Bax,Bcl-2 mRNA and protein expressions (F =20.51,7.59,15.38,11.97,P ＜ 0.05).Conclusion The SNP can induce apoptosis of AC16 cardiomyocyte;selenium combined with SNP has an interactive effect on AC16 cardiomyocyte,indicating that selenium has protective effect on NO modiated myocardial apoptosis.

Objective:To investigate the relationship between single nucleotide polymorphisms of transforming growth factor-β2 (TGFβ2) gene and Keshan disease (KD) in Han population of Shaanxi Province.Methods:KD region in Huangling County, Yan'an City, Shaanxi Province was selected as the investigation site in this study. Using the method of cluster random sampling, 52 families with KD in 6 administrative villages in Huangling County (Duanjiawan Village, Taoqu Village, Yaoping Village, Jianzhuang Village, Anjiao Village in Yaoping Town, and Houziping Village in Diantou Town) were selected for epidemiological investigation. According to the "Diagnosis of Keshan Disease" (WS/T 210-2011), 285 subjects were identified, including 79 patients with KD (case group) and 206 healthy controls (control group). Genomic DNA was extracted from the peripheral venous blood. The polymorphism of genetic variation of TGFβ2 gene rs6658835 was genotyped by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF). Chi-square (χ 2) test and t-test were used to analyze the baseline data, and binary logistic regression model was used to analyze the influencing factors of KD, all samples were tested for Hardy-Weinberg equilibrium using goodness-of-fit χ 2 test, differences in genotype and allele frequencies between case and control groups were compared by χ 2 test, and logistic regression analysis was used to compare the genotype frequencies between two groups after adjusting for confounding factors. Results:Epidemiological investigation showed that there were significant differences in age and heart murmur between case group and control group ( t = 7.03, χ 2 = 9.66, P < 0.05). The analysis of binary logistic regression model showed that the influence of age on KD was statistically significant (χ 2 = 20.72, P < 0.001). The gene frequency distribution of TGFβ2 gene rs6658835 in case group and control group conformed to the Hardy-Weinberg equilibrium (χ 2 = 0.02, P = 0.900). Correlation analysis results: the difference of genotype frequency of TGFβ2 gene rs6658835 in case group (GG, GA, AA: 6.3%, 38.0%, 55.7%) and control group (GG, GA, AA: 10.7%, 43.7%, 45.6%) was not statistically significant (χ 2 = 2.78, P = 0.249). After adjustment by age, the difference of genotype frequency and dominant model of TGFβ2 gene rs6658835 in case group and control group was statistically significant (χ 2adj = 5.43, 4.86, P < 0.05), the difference of recessive model of TGFβ2 gene rs6658835 in case group and control group was not statistically significant (χ 2adj = 2.12, P = 0.145). Conclusion:TGFβ2 gene rs6658835 is associated with KD in Han population of Shaanxi Province.

Objective To investigate the relationship between single nucleotide polymorphisms of Bcl-2 related anti apoptotic protein 3 (BAG3) gene and Keshan disease (KD) in north Chinese Han population.Methods In 2002 a total of 285 Chinese Han subjects,including 79 KD patients and 206 control subjects were involved in this study.Genomic DNA was extracted from the peripheral venous blood sample.Blood samples were provided by the Institute of Endemic Disease Prevention,Xi'an Jiaotong University,and stored at 80 ℃.The polymorphism of genetic variation was genotyped by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF).The data was analyzed using TYPER 4.0 or SPSS16.0 software.All sample groups were tested for Hardy-Weinberg equilibrium using goodness-of-fit x2 test.Differences in genotype distribution and allele frequencies between case and control were compared by x2 test.Logistic regression analysis was applied to detect association using age as a confounding factor.Results All sample group passed the Hardy-Weinberg equilibrium test (P ＞ 0.05).Significant differences were not observed in genotype distribution between cases (rs2234962:CC,CT,TT were 0.0％,0.0％ and 100.0％,respectively;rs196295:GG,GA,AA were 22.8％,54.4％ and 22.8％,respectively;rs3858339:GG,GT,TT were 5.1％,38.0％ and 56.9％,respectively;rs3858340:TT,TC,CC were 5.1％,38.0％ and 56.9％,respectively) and controls (rs2234962:CC,CT,TT were 0.0％,1.0％ and 99.0％,respectively;rs196295:GG,GA,AA were 21.4％,51.5％ and 26.2％,respectively;rs3858339:GG,GT,TT were 5.8％,34.5％ and 59.7％,respectively;rs3858340:TT,TC,CC were 5.8％,34.5％ and 59.7％,respectively) for rs2234962,rs3858339,rs196295 and rs3858340 on BAG3 gene (x2 =0.685,0.408,0.330,0.330,P ＞ 0.05).Significant differences were not observed in genotype after agecorrecting between cases and controls for 4 SNPs on BAG3 gene (x2 =0.001,0.019,1.009,0.019,P ＞ 0.05).Conclusion The results suggest that the BAG3 gene might not be a susceptibility gene of KD in north Chinese Han population.

Objective To investigate the relationship between single nucleotide polymorphisms of interleukin 23 receptor (IL-23R) gene and Keshan disease (KD) in Northwest Chinese Han population.Methods A total of 285 Chinese Han subjects from Huangling,Shaanxi,including 79 KD patients (case group) and 206 control subjects (control group) were involved in this study.Genomic DNA was extracted from peripheral venous blood.The polymorphism of genetic variation was genotyped by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF).All sample groups were tested for Hardy-Weinberg equilibrium using goodness-of-fit x2 test.Differences in genotype distribution between two groups were compared by x2 test.Logistic regression analysis was applied to detect association using age as a confounding factor.Results The gene frequency distribution of IL-23R gene rs10889677 in case group and control group conformed to the Hardy-Weinberg equilibrium (x2 =0.254,P ＞ 0.05).Correlation analysis results:the difference of genotype frequency of IL-23R gene rs10889677 in case group (CC,CA,AA were 6.3％,36.7％,57.0％,respectively) and control group (CC,CA,AA were 5.3％,43.2％,51.5％,respectively) was not statistically significant (x2 =1.008,P ＞ 0.05).After age adjustment,there was no significant difference in genotype frequency of IL-23R gene rs10889677 (x2sdj =0.669,P ＞ 0.05) between two groups.Conclusion There is no correlation between IL-23R gene rs10889677 and KD in Northwest Chinese Han population.

Objective:To screen differential metabolites and metabolic pathways in urine of adult patients with Kashin-Beck disease (KBD), so as to provide scientific basis for finding specific biomarkers and pathogenesis of KBD.Methods:In Yongshou County, the KBD area in Shaanxi Province, adult KBD patients were selected as the case group, and healthy people without clinical symptoms of KBD were selected as the control group in the same disease area. The subjects' fasting mid-morning urine was collected, and liquid chromatography-mass spectrometry (LC-MS) technology was used to detect small-molecule metabolites in the urine. Multivariate statistical analysis [partial least square discriminant analysis (PLS-DA)] and comparison with KEGG and human metabonomics database (HMDB) were used to identify and screen differential metabolites and metabolic pathways in KBD patients.Results:A total of 58 subjects were included, 39 cases in the case group, including 23 males and 16 females; the age was (61.2 ± 7.8) years old; the body mass index was (22.7 ± 6.5) kg/m 2. There were 19 cases in the control group, including 10 males and 9 females; the age was (50.0 ± 9.0) years old; the body mass index was (24.3 ± 5.5) kg/m 2. Three first-order differential metabolites (HT-2 toxin, T-2 tetraol and seleno-adenosine selenomethionine) were identified and screened, which were highly related to the pathogenesis of KBD, and all were down-regulated. There were 38 second-order differential metabolites, among them, 10 were up-regulated and 28 were down-regulated. Nine differential metabolic pathways were screened, mainly involving amino acid metabolism, lipid metabolism and energy metabolism. Conclusions:The urine metabolism profiles of adult KBD patients and healthy people are significantly different, mainly involving amino acid metabolism, lipid metabolism and energy metabolism. The first-order differential metabolites HT-2 toxin, T-2 tetraol and seleno-adenosine selenomethionine are highly correlated with the pathogenesis of KBD.