Aim To evaluate the effect of CYP1A1 on intracellular Ca2+ in human vascular endothelial cells(HVEC304).Methods The expression plasmid of human CYP1A1 was constructed,HVEC304 was transfected,and the intracellular calcium ion level was monitored by microfluorescent technique.Result The expression plasmid of human CYP1A1 was constructed and its expression location was extranuclear.In HVEC304 cells over-expressing CYP1A1(HVEC304-CYP1A1),Ca2+ basal level decreased obviously(P

Objective To investigate the effect of advanced glycation end products (AGEs) on the disruption of tight junctions in rat glomerular endothelial cells (rGEnCs) and the role of renin-angiotensin system (RAS) in this pathological procedure.Methods Primary cultured rGEnCs were incubated with AGEs at concentrations of 20 mg/L,40 mg/L and 80 mg/L,for 6 h,12 h and 24 h respectively.The cells were treated with captopril (1 mmol/L) or valsartan (10 μ mol/L)to block RAS.The endothelial permeability was investigated by transendothelial electrical resistance and the flux of fluorescein isothiocyanate-conjugated bovine serum albumin.The expression of AGEs receptor (RAGE),tight junction proteins [occludin,claudin-5,junctional adhesion molecules A (JAM-A) and zona occludens-1 (ZO-1)]and RAS components [angiotensinogen,renin and angiotensin Ⅱ type 1 receptor (AT1)]were detected by Western blotting.Immunofluorescence was used to demonstrate the disruptions of the tight junction proteins.The activity of angiotensin converting enzyme (ACE) was evaluated by UV spectrophotometry.Angiotensin Ⅱ (Ang Ⅱ ) was measured by enzyme immunoassay.Results The monolayer permeability,the expression of RAGE,the activity of ACE,the concentration of Ang Ⅱ and the expression of AT1 of rGEnCs were increased after induced by AGEs.Meanwhile,AGEs decreased the expression of occludin,claudin5 and JAM-A and induced disruption of tight junction proteins.Pretreatment with anti-RAGE antibody (100 mg/L),captopril or valsartan could attenuate the detrimental effect of AGEs.Conclusion The changes of permeability induced by AGEs in glomerular endothelial cells are partly mediated by RAS through RAGE.

Objective To explore the features and influencing factors of ambulatory blood pressure in chronic kidney disease (CKD) patients.Methods A total of 540 CKD patients from May 2010 to May 2012 in our department were enrolled in this study.Ambulatory blood pressure monitoring was carried out.Blood pressure (BP),proteinuria and other clinical parameters were measured regularly.Ultrasonography was used to evaluate cardiac structure and function,carotid intima-media thickness and plaque.Univariate and multivariate analysis were used to examine the association between BP and clinical parameters.Results 63.9％ of CKD patients was non-dipper BP pattern,and 36.1％ was dipper BP pattern.As compared to dipper BP patients,those with non-dipper BP had higher ratio of nighttime/daytime proteinuria (0.51±0.29 vs 0.42±0.21,P ＜ 0.01),lower estimated glomerular filtration rate (eGFR) [(56.2±48.2) vs (75.5±56.5) ml· min 1 · (1.73 m2)-1,P ＜ 0.01],higher serum cystatin C[(2.8±2.0) mg/L vs (2.1±2.0) mg/L,P ＜ 0.01],higher left ventricular mass index [(53.7±23.2) vs (45.1± 16.3) g/m2,P ＜ 0.01] and severely damaged left ventricular diastolic function and higher carotid intima-media thickness [(0.7±0.3) vs (0.6±0.2) mm,P＜ 0.01].Nighttime blood pressure was independent predictor for proteinuria,eGFR and left ventricular mass index.Conclusions Nondipper blood pressure pattern is very common in CKD patients.Nighttime pressure is closely associated to renal damage and cardiovascular injuries.

Objective To evaluate the applicability of glomerular filtration rate (GFR) caJcuated by Cock-croft-Gault equation, MDRD1 equation and abbreviated MDRD equation in the chronic glomerulonephritis. Methods 143 chronic kidney disease(CKD) patients diagnosed in our hospital from January 2005 to December 2007 were investigated. Glomerular filtration rate(GFR) was estimated with Cockcroft-Gault equation, MDRD1 equation and ab-breviated MDRD equation. The accuracy of estimated GFRs was compared with 99Tcm-DTPA-GFR in chronic glomer-ulonephritis patients. Results Bland-Altman analysis demonstrated that Cockcroft-Gault equation was more consist-ent with GFR and sGFR than the other equations. Linear regression analysis showed that the slopes of Cockcroft-Gault equation were closer to the identical line,with a smaller bias and higher accuracy . When compared, the bias as well as accuracy of estimated GFRs with sGFR in different stages of CKD,Cockcroft-Gault equation showed good results. Condusion When plasma creatinine is checked with enzymatic method, Cockcroft-Gault equation is more appropriate than MDRD1 equation and abbreviated MDRD equation for estimating renal function in chronic glomeru-lonephritis patients.

Objective To investigate the effects of supernatant of cultured mesangial vcells with serum IgA1 from [gA nephropathy patients on apoptosis of podocyte. Methods Jacalin affinity chromatography and Sephacryl S-200 molecular sieve chromatography were used to isolate IgA1. Apoptosis rate of podocyte was assessed by flow cytometer. Monomeric IgA1 (mIgA1) was transformed to aggregated IgA1(aIgA1) by heating. IgA-mesangial cell supernatant was prepared by collecting spent medium in which growth-arrested mesangial cells were incubated with different aIgA1, then the medium with RPMI 1640 containing 0.5%FBS was cultured with growth-arrested podocyte. Real time PCR was used to detect the mRNA expression of Bcl-2, Bax, Fas and Fas-L. Results Apoptosis rate of podocyte by supematant of cultured mesangial cell with algal from IgAN patients was higher than that from healthy and control groups [(28.5±5.9 ) % vs (22.5± 5.8)%, (20.5±4.5)%, all P<0.05]. Fas mRNA expression of podocyte exposed to supematant of cultured mesangial cells with aIgA1 from IgAN patients increased significantly and was 1.89 folds of control (P<0.05), while Bcl-2 mRNA expression significantly decreased and was 72% of control (P<0.05). The concentrations of Ang Ⅱ and TGF-β in supernatant of cultured mesangial cells with IgA1 from IgA nephropathy were significantly higher than those from healthy control [(13.2±3.4) ng/L vs (8.2±2.3) ng/L, /'<0.05; (15.4±3.4) ng/L vs (10.8±3.2) ng/L, P<0.05]. Conclusion Supernatant of cultured mesangial cells with IgA1 from IgA nephroapthy patients can induce apoptosis of podocyte, which may play a role in the progression of IgAN.

ObjectiveTo evaluate the applicability of glomerular filtration rate estimation equations in elderly Chinese patients with chronic kidney disease(CKD). MethodsA total of 103 elderly CKD patients were investigated. Glomerular filtration rate (GFR) was estimated by Coekcroft -Gault equation, MDRD1 equation, abbreviated MDRD equation, Jellife 1973 equation, Mawer equation, Hull equation, Jellife 1971 equation, plasma creatinine reciprocal equation, Gate equation and Bjornsson equation. The accuracy of estimated GFRs was compared with that of GFR estimated by 99mTe-DTPA-GFR (sGFR)in elderly CKD patients. ResultsBland-Altman analysis demonstrated that GFRs evaluated by Cockcroft -Gault equation, Bjornsson equation and Hull equation were more consistent with sGFR than the other equations. But all the equations were not well consistent with sGFR. Linear regression showed that the slopes of Jellife 1973 equation and Cockcroft-Gault equation were closer to the identical line. Bias of MDRD1 equation was smaller. Accuracy of Bjornsson equation, Jellife 1973 equation and Cockcroft-Gault equation were higher than the other equations. When comparing the bias as well as accuracy of estimated GFRs with sGFR in different stages of CKD, GFR estimated by Bjornsson equation, Coekcroft-Gault equation and Jellife1973 equation showed better Results . ConclusionsWhen plasma creatinine is checked by enzymatic method, GFR estimating equations may show great bias in elderly Chinese CKD patients.

Objective To explore the effect of high glucose on the renin-angiotensin system (RAS) in rat glomerular endothelial cells and its probable mechanism.Methods Rat glomerular endothelial cells were stimulated by culture medium containing 5 mmol/L or 30 mmol/L glucose (with or without pre-treatment of captopril and chymostatin) for 12 h,24 h,48 h,72 h.ELISA,real-time PCR,Western blotting and confocal immunofluorescence microscopy were employed to examine the follows:the angiotensin Ⅱ (Ang Ⅱ ) concentration in cell lysate and the culture medium; the mRNA levels of angiotensinogen and renin; the protein levels of angiotensin Ⅱtype 1 receptor (AT1R),angiotensin Ⅱ type 2 receptor (AT2R),renin and angiotensinogen in cell lysate; localization of intracellular AT1R,AT2R and renin.Results Exposure to high glucose for only 12 h or 72 h resulted in a significant increase of Ang Ⅱ levels in the culture medium compared with control cells (P＜0.05),but only exposure to high glucose for 72 h resulted in a significant increase of Ang Ⅱ levels in the cell lysate compared with control cells (P＜0.05).However,exposure to high glucose for 24 h or 48 h had no effects on Ang Ⅱ levels in the cell lysate or culture medium.The captopril and chymostatin were able to antagonize high glucose induced Ang Ⅱ generation when exposure time was 72 h but not 12 h.Exposure to high glucoseincreased the mRNA level of angiotensinogen,but reduced the level of renin mRNA,meanwhile angiotensinogen protein increased,and AT1 protein reduced,but protein levels of AT2R and renin were unchanged.However,the transformation of AT2R from the cell nucleus to cytoplasm was observed.Conclusions High glucose can activate the local renin-angiotensin system in rat glomerular endothelial cells,and the probable mechanism may contribute to ACE and non-ACE pathways.The effects of high glucose on glomerular endothelial cells may also involve in the substrate and receptors of Ang Ⅱ.

Objective To investigate the related factors and prognosis of pulmonary hypertension (PAH) in hemodialysis (HD) patients for early diagnosis and intervention of PAH.Methods A retrospective cohort study was conducted in 183 long-term hemodialysis patients with complete follow-up data from January 1,2010 to December 30,2015 from the blood purification center of the Third Affiliated Hospital of Sun Yat-sen University.The follow-up deadline was December 30,2017,and the endpoints were death and cardiovascular events.The clinical data,laboratory examinations,cardiac color Doppler ultrasound parameters and prognosis of patients with and without PAH were compared.Multivariate logistic regression was used to analyze the risk factors for PAH in HD patients.The survival rates were calculated by Kaplan-Meier method,and the survival curves were compared by Log-rank test between the two groups.A multivariate Cox proportional hazard regression model was used to examine the association between PAH and all-cause mortality in HD patients.Results Of the 183 hemodialysis patients,79(43.2％) were female,104(56.8％) were male,and the age was (56.1±16.9) years,of which 72(39.3％) were complicated with PAH.Compared with the non-PAH group,patients in the PAH group was older and had a longer duration of dialysis (both P ＜ 0.05).The left atrial diameter (P=0.002) and the proportion of valvular calcification (P=0.004) were significantly higher in the PAH group than that in the non-PAH group.Logistic regression analysis showed increased age (OR=1.027,95％ CI 1.001-1.053,P=0.041) and increased duration of dialysis (OR=1.129,95％ CI 1.004-1.269,P=0.042) were risk factors for PAH in HD patients.After a median follow-up of 27.8 months,Kaplan-Meier survival analysis showed that all-cause mortality was higher in the PAH group than that in the non-PAH group ~x2=5.636,P=0.018).The main cause of death in two groups was cardiovascular event.Afteradjusting for age,diabetes mellitus,duration of dialysis,valvular calcification,and hypertension,Cox regression showed that PAH increased the risk of all-cause mortality in HD patients (HR=1.894,95％ CI 1.083-3.313,P=0.025).Conclusions HD patients complicated with PAH are more common and the prognosis is poor.Increased age and increased duration of dialysis may be risk factors for PAH in HD patients.Regular color Doppler echocardiography is helpful for early detection and diagnosis of PAH.

Objective: To analyze the relationship between serum uric acid (SUA) level and clinical indicators in maintenance hemodialysis (MHD) patients, and explore its influence on all-cause mortality and cardiovascular mortality. Methods: This study was a retrospective cohort study. Patients who received MHD from the blood purification center of the Third Affiliated Hospital of Sun Yat-sen University from January 1, 2011 to December 30, 2015 were enrolled in the queue. They were divided into 3 groups according to the first and third quantile of the SUA level quartiles, and the baseline data of clinical and laboratory examinations were compared. The correlation between SUA level and clinical indicators was analyzed by Pearson correlation coefficient. Kaplan-Meier method and Cox proportional hazard regression model were used to examine the association between SUA and all-cause mortality and cardiovascular mortality in MHD patients. Results: A total of 201 patients were enrolled in the study. The age of the patients was (56.9±16.7) years and the baseline SUA level was (531.1±137.9) μmol/L. Patients were divided into 3 groups with the first quantile (442 μmol/L) and the third quantile (620 μmol/L) of the SUA quartiles as the boundary points: group 1 (SUA<442 μmol/L, n=52), group 2 (SUA 442-620 μmol/L, n=101) and group 3 (SUA>620 μmol/L, n=48). The results showed that the patients in group 1 were older and had more proportion of patients with diabetes mellitus and cardiovascular diseases than those in group 3 (all P<0.05). Compared to group 3, the serum albumin, serum phosphorus and serum creatinine were lower in group 1, while the hypersensitive C-reactive protein was higher (all P<0.05). Pearson correlation analysis showed that SUA level was positively correlated with albumin (r=0.135, P=0.047), blood phosphorus (r=0.269, P<0.001) and serum creatinine (r=0.333, P<0.001), and negatively correlated with hypersensitive C-reactive protein (r=-0.216, P=0.002). After a median follow-up of 49.8 months, 66(32.8%) all-cause deaths and 32(15.9%) cardiovascular deaths were recorded. Kaplan-Meier method showed that with the decrease of SUA, all-cause mortality (Log-rank χ²=18.27, P<0.001) and cardiovascular mortality (Log-rank χ²=15.04, P=0.001) increased. After adjusting for age, gender, comorbidity and other factors using the Cox proportional hazards model, the all-cause mortality and cardiovascular mortality decreased by 20.1% (HR=0.799, 95% CI 0.651-0.980, P=0.031) and 29.6% (HR=0.704, 95% CI 0.524-0.946, P=0.020) for each 100 μmol/L increase in baseline SUA. Compared to group 1, all-cause mortality (HR=0.332, 95%CI 0.142-0.774, P=0.011) and cardiovascular mortality (HR=0.140, 95%CI 0.030-0.657, P=0.013) were lower in the group 3. Conclusion Low SUA level increases the risk of all-cause mortality and cardiovascular mortality in MHD patients.

Objective To cxplore the optimal levels of serum calcium,phosphorus and intact parathyroid hormone (iPTH) in peritoneal dialysis (PD) patients.Methods This study is a single center,retrospective cohort study.The associations between serum calcium,phosphorus and iPTH and all-cause mortality in 217 PD patients were analyzed.All patients started PD between January 1,2008 and April 30,2016 were enrolled and followed up to December 31,2016.At baseline and every 3 months,biochemical and therapeutic information was collected.Cox proportional hazard regression models and cubic splines analysis were employed to assess the lowest mortality risk ranges in serum markers of bone metabolism.Results There was no significantly difference between patients within target ranges based on KDOQI or KDIGO guideline and those outside the target ranges by Kaplan-Meier survival analysis.The lowest mortality risk ranges were 2.17-2.40 mmol/L for serum calcium,1.20-1.67 mmol/L for serum phosphorus and 180-350 ng/L for serum iPTH by using Cox models and cubic splines analysis.Moreover,cumulate survival had significant difference between patients within the descriptive ranges and those out of the descriptive ranges at time-averaged values but not at baseline values.Conclusions The optimal time-averaged ranges of PD patients are 2.17-2.40mmol/L for serum calcium,1.20-1.67 mmol/L for serum phosphorus and 180-350 ng/L for serum iPTH.These ranges need further validation by large population studies to further conform.