| Objective:1.To analyze the effect of RXR in proliferation inhibition of human neuroblastoma induced by BDE-47;further explore the effect of cell nuclear receptors that taking RXR? as the hub in BDE-47-induced neurotoxicity for uncovering the molecular mechanisms of PBDEs neurotoxicity mediated by unclear receptors.2.To investigate and analyze the abnormal proportion of thyroid functions and the prevalence of thyroid dysfunction in a community health center from Shenzhen.Methods:1.To construct RXR-knockout SK-N-SH and RXR-overexpression SK-N-SH.The RXR-knockout SK-N-SH(RXR/KO-SK-N-SH)were constructed by knockout of RXR genes with CRISPR-cas9 and screening and identification with G418.The RXR-overexpression SK-N-SH cells(RXR/OE-SK-N-SH)were constructed by constructing eukaryotic expression vector and transfecting SK-N-SH with nuclear transfection technique,and screening and identification with G418.The wild-type SK-N-SH,RXR/KO-SK-N-SH and RXR/OE-SK-N-SH exposed to BDE-47 with different concentrations were measured with CCK-8 assay.The expression level of genes RXR?,TR?,TR?,PPAR?,PPAR? in the three SK-N-SH treated with BDE-47 were assayed by q-PCR and Western Blot.2.100 individuals from a community health center of Futian district of Shenzhen city were recruited in this research.The total 8 index of sex,age,height,weight,waist circumference,iodine salt intake,living time in Shenzhen and educational level were survived.The thyroid function indicators in serum was detected by chemiluminescence method,and the B-scan ultrasonography was applied to thyroid morphology check.EpiData 3.1 was used for date input and all statistical analyses were performed using SPSS Statistics(version 22.0,IBM Inc.,Chicago,IL,USA).The baseline characteristics were presented as frequencies with percentages for categorical variables and as mean ± standard deviation(SD)for continuous variables.?2 test was used to compare the abnormal rates of levels of thyroid stimulating hormone(TSH),total T3(TT3),free triiodothyronine(FT3),total T4(TT4),free thyroxine(FT4)in serum and B-scan ultrasonography of thyroid,and the difference of abnormal rate of TSH,TT3,TT4 in serum and B-scan ultrasonography of thyroid in sex,age,iodine salt intake,body mass index(BMI),waist circumference,living time in Shenzhen and education level.The incidence ratios of subclinical hyperthyroidism,hyperthyroidism,subclinical hypothyroidism and hypothyroidism and abnormal ratios of B-scan ultrasonography of thyroid combined with above indicators were further assessed.A univariate logistic regression analysis was performed to assess the factors which may influence on abnormal of B-scan ultrasonography of thyroid.Taking P value less than 0.20 as the standard of potential covariates,a multivariate logistic regression analysis was performed to assess the association between abnormal of B-scan ultrasonography of thyroid with the TSH,TT3,FT3,TT4 and FT4 in serum and other demography parameters.P<0.05(bilateral)was considered the difference statistically significant.Results:1.Successfully constructed RXR-knockout SK-N-SH(RXR/KO-SK-N-SH)and RXR-overexpression SK-N-SH(RXR/OE-SK-N-SH).protein expression of RXR? in RXR/KO-SK-N-SH and RXR/OE-SK-N-SH was 0.13 and 1.87 times higher than that in wild-type SK-N-SH,respectively.2.BDE-47 inhibited the proliferation of wild-type SK-N-SH?RXR/KO-SK-N-SH and RXR/OE-SK-N-SH,significantly,and the inhibition rate increased with the increase of BDE-47 treatment concentration and treatment time.the IC50 of RXR/KO-SK-N-SH is much lower than that of wild-type SK-N-SH and RXR/OE-SK-N-SH and is half of that of wild-type SK-N-SH.3.The mRNA and protein expression of RXR?,TR?,TR?,PPAR? and PPAR? were different in all three cells.RXR? was not expressed in the RXR/KO-SK-N-SH.The mRNA and protein expression of RXR? increased significantly in wild-type SK-N-SH and RXR/OE-SK-N-SH after the three cells treated by BDE-47.TR? and PPAR? expression increased in the wild-type SK-N-SH and RXR/OE-SK-N-SH,but decreased in the RXR/KO-SK-N-SH.With the increase of BDE-47 treatment concentration,the expression of TR? and PPAR? did not change significantly in all three cells.4.The number of abnormal serum FT3 and FT4 levels was 1 among 100 recruited persons in the social health center.Ate iodized salt,iodine-free salt,and the composition ratio of the two alternate eaters were 82%,15% and 3% and the abnormal individual number of serum TSH levels were respectively 17,2 and 7 in the three groups.The abnormal rates of serum TT3 levels were statistically significant among different age groups(P <0.05).The composition ratios of the study population in age groups of less than 26,27 to38,39 to 50,51 to 62 and more than 63 were 7%,51%,22%,12% and 8%,respectively 5,19,14,1 and 2 cases were detected in each group.The abnormal rate of serum TT3 level was statistically significant in different age groups(P<0.05).There was 8.33% in the 51~62 age group and the abnormal rate was the lowest in all age groups,and there were respectively 71.43%,37.25%,63.64% and 75.00% in age groups of less than 26,27 to38,39 to 50 and more than 63.The abnormal rate of serum TSH,TT3,TT4 level and B-scan ultrasonography of thyroid was higher in women than men,but the difference was not statistically significant,and the four indexes were not statistically different in the BMI,waist circumference,Shenzhen residence time and education level group.A total of four cases of subclinical hyperthyroidism,hyperthyroidism and subclinical hypothyroidism were detected,including one case of subclinical hyperthyroidism and one case of hyperthyroidism,two cases of subclinical hypothyroidism and no hypothyroidism.T he number of abnormal of B-scan ultrasonography of thyroid was thirty-three.All of these thyroid dysfunctions were accompanied abnormal of B-scan ultrasonography of thyroid.A univariate logistic regression analysis found that BMI was the influencing factor of abnormal of B-scan ultrasonography of thyroid(95% ci : 1.01~1.30;p ci :1.01~0.05).Multivariate logistic regression did not find an association between thyroid-related hormone abnormalities and thyroid abnormality rate of B-scan ultrasonography.Conclusion:1.RXR? played a critical role in improving cells viability.RXR? affected the expression of TR? and PPAR? in the SK-N-SH treated with BDE-47.BDE-47 toxic effects in the SK-N-SH were performed via activating RXR? and further affecting expression of TR? and PPAR?.2.Subclinical hyperthyroidism,hyperthyroidism and subclinical hypothyroidism patients were found among the 100 recruited persons in the social health center,and hypothyroidism patients were not detected.All patients with thyroid dysfunction were accompanied by thyroid abnormality of B-scan ultrasonography.The serum TT3 level was the lowest in the 51~62 age group and the remaining age groups are higher.There was no association between thyroid hormone level turbulence and thyroid abnormality rates of B-scan ultrasonography in this survey study. |
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