ABSTRACT
Objective To explore the effects of chronic sodium arsenic (NaAsO2) exposure on energy metabolism in human keratinocytes (HaCaT).Methods Malignant transformation was induced when HaCaT cells were continuously exposed to an environmentally relevant level (0.1 μ mol/L) of inorganic arsenic for 28 weeks,HaCaT and malignantly transformed cells were kept in Group of Chronic Disease and Environmental Genomics,School of Public Health,China Medical University.Basal oxygen consumption rate and extracellular acidification rate in control group and malignantly transformed group were measured by Agilent Seahorse XF24.Changes of glucose intake in malignantly transformed HaCaT cells were detected by 2-NBDG.Cells in logarithmic growth phase were collect,mRNA levels of key components in glycolytic pathwa [glucose-6-phosphate dehydrogenase (G6PD),phosphofructokinase (PFKL),glyceraldehyde-3-phosphate dehydrogenase (GAPDH)] and mitochondrial respiratory chain complex [NADH dehydrogenase iron-sulfur protein subunit 3 (NDUFS3),succinate dehydrogenase iron-sulfoprotein subunit (SDHB),cytochrome C (CYTC),CYTC oxidase subunit Ⅳ (COX4L1),ATP synthase F1 subunit (ATP5B),and ATP synthase F0 subunit (ATP5G1)] were determined by real-time quantitative PCR (RT-qPCR).Results The basal oxygen consumption rate of malignantly transformed group [(44.784 ± 10.159) pMoles/min] was lower than that of control group [(66.842 ± 15.756) pMoles/min,t =4.914,P ＜ 0.05].The glucose intakes of malignantly transformed group (1.592 ± 0.410 for cells cultured without serum,and 1.631 ± 0.323 for cells cultured with serum) were significantly higher than those of control group (1.000 ± 0.334,1.000 ± 0.196,t =2.916,4.068,P ＜0.05).The mRNA levels of G6PD (0.555 ± 0.009),NDUFS3 (0.623 ± 0.031) and SDHB (0.702 ± 0.094)in malignantly transformed group were significantly lower than those of control group (1.000 ± 0.027,1.000 ±0.034,1.000 ± 0.114,t =23.690,11.340,2.814,P ＜ 0.05);the mRNA levels of PFKL (1.787 ± 0.176),GAPDH (1.466 ± 0.111),CYTC (2.461 ± 0.179),ATP5B (1.956 ± 0.161),and ATP5G1 (2.055 ± 0.052) were significantly higher than those of control group(1.000 ± 0.153,1.000 ± 0.069,1.000 ± 0.030,1.000 ± 0.091,1.000 ± 0.237,t =4.696,4.985,11.540,7.436,6.021,P ＜ 0.05).Conclusion When continuously exposed to an environmentally relevant level of inorganic arsenic,the glucose metabolic pattern of HaCaT cells is changed along with lower oxygen consumption and higher glycolysis levels.
ABSTRACT
Objective To study the emmetropization in guinea pig eyes during the normal development from birth in order to provide theoretical basis to use guinea pigs as model for research on near-sightedness.Methods Sixty-four guinea pigs were assigned to 8 groups (n=8,4 male/4 female).Each group underwent a series of ocular examinations at one of the 8 time-points (0,1,2,3,5,7,9 and 11 weeks),including refraction(R),radius of corneal curvature (RCC),depth of anterior segment(AS),thickness of crystalline lens (CL),length of vitreous chamber (VC)and axial length (AL).Pooled results from both eyes of the same animals with mixed sexes can be used for further assessment of the emmetropization with the associated biometrical changes of the eye.Correlations were made between RCC,AS,CL,VC,AL and R. Results There were no significant differences between the right eye and left eye or between male and female in results of all the examinations. The refraction at birth was (+5.25?0.22 )D in guinea pigs.This value rapidly decreased during the first 3 weeks,approaching(+1.34?0.61) D by 11 weeks.There was no significant difference in refraction between 9 and 11 weeks (P= 0.215). The RCC was (3.23?0.01) mm at birth,AS was (1.20?0.00) mm at birth,CL was (2.72?0.02) mm at birth and VC was (3.27?0.01) mm at birth,they increased within the first 3 weeks despite a transient decrease in the RCC within the first week.Such an increase continued except the AS which became constant after 3 weeks.The VC was more correlated to the emmetropization (r=-0.818,P
ABSTRACT
By using antr-ras P21 mouse monoclonal (MoAb), SCI-Oncogema 1, the authors examined immunohistochemica! staining in pancreatc adeocarcinoma. The percentage of positive staining was 24/43 (558%). Furthermore it was indicated that the positive staining rate of antr-ras P21 MoAb was related to either histopathological grade or clinical stage. Upon statistical analysis of the correlation between the staining of anti-ras P21 and patient prognosis with Kaplan-Meier curve and Log-rant test, the positive staining cases showed comparatively better prognosis than the negative ones. Our study suggests that ras P21 expression may be important in the early stage of pancreatic carcinoma.