Objective To investigate the inhibitory effect of blocking PI3K/AKT pathway by wortmannin on hypoxia-inducible factor 1α (HIF-1α) and the effect on the expression of glycolysis associated genes in human esophageal carcinoma cell lines TE1 and TE13,and to analyze the relation between PI3K/AKT-HIF-1α pathway and glycolysis in esophageal carcinoma cells. Methods Esophageal carcinoma cell lines TE1 and TE13 pretreated with wortmannin (2 μmol/L) were incubated under normoxic and hypoxic conditions.And each cell line was divided into four groups.The expression of HIF-1α and glycolysis associated genes GLUT-1,LDHA and HK-Ⅱ at protein level were measured by.Western blot.The expression of HIF-1α,GLUT-1,LDHA and HK-Ⅱ at mRNA level was determined by real-time PCR. The activities of LDH and HK-Ⅱ and lactic acid (LA)concentration in the culture supernatant were tested with spectrophotometer method.Results Under normoxic condition,HIF-1α was expressed in TE1 cells and the expression of HIF-1α was inhibited by wortmannin (2 μmol/L),the most significant inhibitory effect was at 12 hours,therefore 12 hours was selected for the subsequent hypoxia experiment.Compared with untreated cells,the expression of HIF-1α、HK-Ⅱ 、GLUT-1、LDH-A at protein level significantly decreased in TE1 and TE13 cells after pretreated with wortmannin (P ＜ 0.05),and the expression of HIF-1α、HK-Ⅱ at mRNA level significantly decreased (P＜ 0.05).Under normoxic and hypoxic conditions,the HK-Ⅱ and LDH activities in TE1 and Te13 esophageal carcinoma cells significantly decreased after treated with wortmannin compared with untreated cells (P＜0.05).Under hypoxia condition,the enzyme activity increased in untreated cells (P＜ 0.05). Under normoxic and hyp0xic conditions,the lactic acid concentration in the culture supernatant obviously decreased in cells treated with wortmannin compared with untreated cells (P＜ 0.05). Under hypoxia condition,lactic acid concentration increased in wortmannin treated cells (P ＜ 0.05). Conclusions Under normoxic and hypoxic conditions,wortmannin decrease lactic acid concentration through inhibiting the expression of HIF-1α and glycolysis associated genes, which indicate PI3K/AKT-HIF-1α pathway was closely related to glycolysis in esophageal carcinoma cells.

Objective To evaluate the effect of cardiovascular disease (CVD) on the healthcare costs associated with chronic kidney disease (CKD).Methods A total of 96 patients with stage 3-4 CKD treated at CKD clinic of Beijing Friendship Hospital,Capital Medical University were enrolled in the study.Their mean age was (61.6±9.5) years including 65 male and 31 female patients.A retrospective analysis of clinical material and health claims between January 2010 and October 2010 was conducted.Firstly,patients were grouped according to median CKD-associated healthcare cost and clinical characteristics were compared between two groups.Secondly,patients were stratified into three categories based on CVD prevalence (with acute cardiovascular events,with CVD but no acute events,and without CVD),and CKD-associated healthcare costs were assessed among the groups.Finally,the potential factors influencing CKD-associated healthcare costs were evaluated by optimal scaling regression analysis.Results During January to October in 2010,median CKD-associated healthcare costs was 13960.5 yuan (interquartile range 10226.5,19667.2 yuan).In the group with higher healthcare costs,more females,greater urine albumin-creatinine ratio,more emergency observations and/or hospitalizations caused by acute cardiovascular events,higher diabetes mellitus prevalence and calcium-phosphorus products,and lower eGFR and hemoglobin levels were found (P＜0.05,respectively).In contrast,the total prevalence of CVD was not significantly different between the groups (P=0.386).When grouping by CVD prevalence,significant differences of CKD-associated healthcare costs were observed only between patients with acute cardiovascular events and the other two groups (P＜0.01,respectively).The median healthcare cost of the former was approximately twice as higher as that of the other two groups,and the maximal cost was also found in the acute-cardiovascular-event group.For the optimal scaling regression analysis,both emergency observations and/or hospitalizations caused by acute cardiovascular events and diabetes mellitus entered the equation,and standardized coefficients were -0.538 and -0.217 respectively (P＜0.01 and P＜0.05).Conclusions Emergency observations and/or hospitalizations caused by acute cardiovascular events are important factors inducing high CKD-associated healthcare costs in patients with stage 3-4 CKD.Therefore,the prevention of acute cardiovascular events may be favorable to reduce CKD-associated healthcare costs.Larger and longer-time perspective studies are required to confirm it.In addition,diabetes mellitus also influences CKD-associated healthcare costs.