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Objective:To evaluate the value of combined measurement of urinary insulin-like growth factor-binding protein 7 (IGFBP7) and urinary metalloproteinase inhibitor-2 (TIMP-2) in the early diagnosis and prognosis of cardiac surgery-associated acute kidney injury (CSA-AKI).Methods:From March 2018 to June 2018, cardiac surgery patients admitted to the cardiac macrovascular surgery department of the First Affiliated Hospital of Nanjing Medical University were prospectively included, and the blood creatinine was monitored to observe the presence of acute kidney injury (AKI). The prognostic information of the patients was collected, including in-hospital dialysis, in-hospital death, complete recovery of kidney function at discharge, death in one year after surgery, and progression to chronic kidney disease. The levels of urine IGFBP7 and TIMP-2 at 6 h, 24 h and 48 h after cardiac surgery were detected by enzyme linked immunosorbent assay (ELISA), and the urine creatinine (Cr) was also measured. Moreover, receiver operating characteristic curves (ROC) were plotted and the areas under the curves ( AUC) were calculated to evaluate the predictive value and prognostic value of urinary [TIMP-2]·[IGFBP7] (T*I for short) and urine T*I/urine Cr 2 in CSA-AKI. Results:A total of 74 patients with age of (58.43±10.91) years old and 47 males, were enrolled in this study, of which 24 cases (32.4%) had AKI and 10 cases (13.5%) had stage 2-3 AKI. Compared with the non-AKI group, the AKI group had significantly higher levels of urine T*I levels at 6 h and 24 h (both P<0.05). The AUC of T*I at 24 h predicting for AKI was 0.71(95% CI 0.59-0.81, P=0.001, cutoff value 0.020, sensitivity 79.2%, specificity 56.0%), while the AUC for stage 2-3 AKI was 0.85 (95% CI 0.75-0.92, P<0.001, cutoff value 0.083, sensitivity 70.0%, specificity 90.6%). Urinary T*I normalized for urinary creatinine excretion did not show better predictive value. In addition, of T*I at 24 h predicting for poor hospitalization outcome, renal recovery, and one year postoperative death, the AUC was 0.82(95% CI 0.71-0.90, P=0.001), 0.80(95% CI 0.66-0.86, P<0.001), and 0.81(95% CI 0.70-0.89, P=0.047), respectively. Conclusion:The combined detection of TIMP-2 and IGFBP7 in urine is expected to be a biomarker for early diagnosis of CSA-AKI and has certain clinical value in predicting the prognosis of CSA-AKI.
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Objective To investigate the urinary iodine level and the thyroid function of early pregnant women in Qingdao,to explore the relationship between the iodine nutritional status and hypothyroidism.Methods 326 early pregnant women (<12 gestational weeks)were collected.The urinary iodine concentration and serum thyroid stimulating hormone (TSH),free thyroxine (FT4 ),thyroid peroxidase antibody (TPOAb)were detected. Results The median of urine iodine of pregnant women was 176.57μg/L,and the ratios of iodine deficient,iodine adequate,iodine more than adequate and iodine excessive groups were 3 8.65%,41.10%,15.95% and 4.30%.The total ratio of innormal urine iodine was 58.9%(192 /326).The differences between the ratios of the iodine deficient group,iodine excessive group and iodine adequate group both had statistically significant(χ2 =4.308,P <0.05;χ2 =4.826,P <0.05 ).There was no effect of iodine intake levels on serum thyroid peroxidase antibody.Conclusion There is no iodine deficiency among early pregnant women in Qingdao,but still individuals expose iodine deficiency.Iodine deficiency and iodine excessive are the risk factors of hypothyroidism during early pregnancy women,so we commend to detect the urinary iodine level.
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Objective To explore the effect and mechanism of fluvastatin on the expression of fibronectin(FN) in human peritoneal mesothelial cells (HPMCs) induced by high-glucose peritoneal dialysate (HGPDS).Methods Cultured HPMCs were randomly divided into control,HGPDS,HGPDS plus GSK650394 10-5 mol/L (the competitive inhibitor of SGK1),different concentrations of fluvastatin,fluvastatin 10-6 mol/L and GSK650394 10-5 mol/L alone.The morphology change of HPMC was observed by light microscopy.The cellular viability was detected by MTT colorimetry.The mRNA and protein expressions of serum and glucocorticoid-inducible kinase 1 (SGK1) and FN were detected by RT-PCR,Western blotting or ELISA.Results After incubation with HGPDS,the cell morphology changed from typical cobblestone-like appearance to fibroblast-like appearance,and the cell viability was inhibited significantly (P＜0.05).Fluvastatin 10-6mol/L and GSK650394 could improved the cell morphology and the cell viability injured by HGPDS (P＜0.05).Compared with the normal control group,the mRNA and protein expressions of SGK1 and FN increased significantly in HPMC treated with HGPDS(P＜0.05).GSK650394 significantly decreased the high expression of SGK1 and FN (P＜0.05),also the fluvastatin had same effects as GSK650394 in dose-dependent manner (P＜0.05).Conclusions High-glucose peritoneal dialysate can increase FN expression in human peritoneal mesothelial cells,which can be attenuated by fluvastatin.The protective role of fluvastatin in HPMC may be partially achieved through the signal pathway of SGK1.
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Objective To investigate the effect of fluvastatin (FLV) on the expression of β1 integrin in puromycin aminonucleoside (PAN)-treated podocytes and its mechanism.Methods Cultured human podocytes were divided into PAN,different concentrations of fluvastatin (1 × 10-8 to 1 × 10-5 mol/L),SOD,H2O21 groups respectively.Expressions of β1 integrin and reactive oxygen species (ROS) in podocytes were detected by Western blotting and DCFHDA (2' 7'-Dichlorofluoresecein 3' 6'-diacetate) respectively.The viability of podocyte was determined by MTT colorimetry.Results PAN and H2O2 significantly decreased the expression of β1 integrin and increased the synthesis of ROS in podocytes (P＜0.05respectively).Lower concentration fluvastatin or SOD treatment up-regulated β1 integrin and downregulated ROS of podocytes induced by PAN (P＜0.05 respectively).MTT revealed that lower podocyte viability was found in higher concentration fluvastatin,PAN and H2O2 groups.Lower concentration fluvastatin and SOD could protect podocytes against PAN.Conclusion Fluvastatin attenuates the injury of podocyte induced by PAN and increases the expression of β1 integrin,whose mechanism may be associated with the inhibition of the ROS activity.
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Objective To explore the effect and associated mechanism of fluvastatin combined with losartan on the expression of vascular endothelial growth factor (VEGF) in human podocytes induced by angiotensin (Ang)Ⅱ.Methods The differentiated human podocytes were cultured with various concentrations of Ang Ⅱ (10-9 to 10-5 mol/L) in vitro,followed by treatment of fluvastatin (10-7,10-6 and 10-5 mol/L),losartan (10-7,10-6 and 10-5 mol/L),extracellular signal-regulated kinase (ERK) inhibitor PD98059,and combination of fluvastatin and lolsartan.Expressions of VEGF and phosphorylation (p)ERK1/2 protein in podocytes were detected by Western blotting.RT-PCR was used to examine VEGF mRNA expression (P＜0.01).Results Ang Ⅱ up-regulated the expressions of VEGF and p-ERK1/2 in time-and dose-dependent manner.Above elevated expressions of VEGF and p-ERK1/2 induced by Ang Ⅱ could be down-regulated by fluvastatin,losartan and PD98059 respectively (P＜0.05).More obvious reduction of above expressions was found in combination of fluvastatin and losartan as compared to single agent (P ＜0.05).Conclusions Either fluvastatin or losartan can down-regulate the over-expression of VEGF and p-ERK1/2 induced by Ang Ⅱ in human podocytes,and their combination has a cooperative effect.The ERK signaling pathway may be one of the mechanisms of their renal protective effects.
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Objective To investigate the clinical and renal pathological features in the new rat model induced by anti-human podocyte-protein antibody. Methods The rat model was induced by once intravenous injection of rabbit anti-human podocyte-protein antiserum which was prepared at first. Male Sprague-Dawley rats (n=36) were randomly divided into six groups (6rats in each group): control group (CG), the time points of day 7 group (D7), day 14 group (D14),day 21 group (D21) and day 28 (D28) group after antiserum injection, and day 28 group after the normal rabbit serum injection (NRG). The level of 24 hour proteinuria, the clearance of creatinine,albumin, blood urea nitrogen, triglyceride, cholesterol and serum creatinine were measured. The renal morphology was detected under the light microscope, immunofluorescence microscope, and electron microscope. Results 24-hour proteinuria (mg) was gradually increased, and the level of proteinuria in D28 (48.56±13.80) was significantly higher than that in CG (5.34±2.77, P＜0.01)and NRG (11.32±4.90, P＜0.01). The clearance of creatinine (ml/min) and serum creatinine (μmol/L) in D28 (0.90±0.47, 33.48±9.94) were significantly different from CG (1.68±0.54, P＜0.05;26.03±2.67, P＜0.05), but showed no difference with NRG (1.34±0.87, P＞0.05; 27.40±4.73, P＞0.05). The level of albumin (g/L) was lower in D7, D14, D21, D28 (28.20±0.87, 27.80±1.97,27.42±1.66, 27.77±1.95) than CG (29.98±0.76, P＜0.05). But there was no difference in the level of albumin among the groups after antiserum injection and NRG (28.68±1.18, P＞0.05). The level of blood urea nitrogen, triglyceride, cholesterol showed no difference among the groups (P＞0.05). The renal morphology showed no obvious changes between CG and NRG. Among the groups after antiserum injection, the renal pathological changes under the light microscope were some spikes formation in D28. Immunostained for rabbit IgG in rat glomeruli progressively decreased over the 28 days, while rat IgG progressively increased. The renal section deposition for rat complement 3 reached a maximum at day 21 then decreased afterward. Under the electron microscope, there were immune complexes and foot process fusion at day 14. Conclusions The new rat model induced by anti-human podocyte-protein antibody showing typically clinical and pathological changes of the membranous nephropathy is successfully established.
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Objective To investigate the role of fibroblast growth factor-23 (FGF23) in secondary hyperparathyroidism (SHPT). Methods (1)Serum FGF23 and intact parathyroid hormone (iPTH)from 38 maintenance hemodialysis (MHD) patients were measured by ELISA and chemiluminescence enzyme immunoassay respectively.(2) Parathyroid cells from six SHPT patients underwent parathyroidectomy with forearm autotrlansplantation were cultured for 24 h,then were induced by 0.1 mg/L FGF23.The supernatant was collected at 0.6,12,24 and 48 h respectively. The concentration of iPTH was measured by chemiluminescence enzyme immunoassay. (3)Protein expression of Klotho,FGFR1,FGFR3,GATA-3 and PCNA in parathyroid tissue from 33 SHPT eases and 3 healthy people were detected by immunohistochemistry SP and PV methods respectively. Positive cell rate and absorbance were calculated. Results (1) Serum FGF23 [(3901.85±2618.11) ng/L] was positively correlated with serum iPTH [(460.00±489.77) ng/L] in MHD patients. (2) 0.1 mg/L FGF23 suppressed iPTH secretion of parathyroid cells only at 24 h time point in vitro (P<0.05). (3) Expression of GATA-3, FGFR3, Klotho and PCNA was significantly increased and FGFRl was signiticantly decreased in parathyroid tissue of SHPT-patients as compared to healthy people. (4) Positive cell rate of GATA-3 was positively correlated with iPTH (r~2=0.1901, P=0.0425) and PCNA (r~2=0.2584, P=0.0025). Klotho was positively correlated with FGFRI and FGFR3 (r~2=0.2046, P=0.0082;r~2=0.2833, P=0.0014). PCNA was negatively correlated with FGFR1 (r~2=0.1292, P=0.0399) and positively correlated with FGFR3 (r~2=0.1226, P=0.0457). FGFR1 was negatively correlated with serum phosphate (r~2=0.2329, P=0.0044) and positively correlated with serum calcium (r~2=0.1422, P=0.0305). Conclusions FGF23 level is positively correlated with iPTH level in MHD patients. FGF23 can inhibit iPTH secretion of parathyroid cells in a weak and short way, which may be associated with the proliferation of GATA-3 positive cells and parathyroid cells, the up-regulation of FGFR3 and the down-regulation of FGFR1 expression.
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Objective To investigate the effect of hyperphosphatemia on type Ⅱa sodium dependent phosphate cotransporter (NaPi-2)mRNA expression in the kidneys of 5/6 nephrectomized (STNx)rats and evaluate the interference of Renagel. Methods Forty-two male Sprague-Dawley rats underwent 5/6 nephrectomy or sham operation. The animals were then divided into seven groups: (1)STNx+high-Pi diet(HP) group (1.2%P); (2)STNx+low-Pi diet(LP) group(0.2%P); (3)STNx+ normal-Pi diet (NP) group; (4)STNx+HP+Renagel-treated (Ren)group; (5)sham+HP group; (6) sham+LP group; (7)sham+NP group. Serum ionized calcium (iCa) , phosphorus, and iPTH levels were detected at day 2,7,14.The kidneys were removed after the rats were sacrificed on day 14 and serum 1,25-(OH)2D3 and FEp were measured. RT-PCR was applied to examine NaPi-2 mRNA. Results Serum P and iPTH levels in STNx+HP group were significantly higher than those in STNx+ LP and 3 sham groups (P0.05)among these five groups mentioned above. Serum P and iPTH levels in Renagel-treated group were much lower than those in STNx+HP group (P
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Objective To investigate the mechanism of high phosphate - stimulated calcification in vascular smooth muscle cell. Methods The cell culture of bovine aortic smooth muscle was performed. Osteocalcin (OC) mRNA expression and calcium deposition in different phosphate concentrations [normal phosphate (Pi 1. 5mmol/L), high phosphate (2.0mmol/L)] were determined by radioimmunoassay, RT-PCR, and 0-cresolphthalein complexone method respectively. Results After 72 hours,high phosphate treatment of smooth muscle cells(SMC) enhanced the expression of osteocalcin protein [Pi 2. 0 mmol/L versus Pi 1. 5 mmol/L: (15. 03?2. 60) pg/u-g protein versus (2. 98 ?0.84) pg/ug protein, P
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AIM To observe the effect of phosphonoformic acid (PFA) in different concentration on vascular calcification induced by elevated phosphate. METHODS The cell culture of bovine aortic smooth muscle were performed. Calcium deposition in different phosphate and PFA concentrations were determined by O cresolphthalein complexone method, and osteocalcin expression by radioimmunity and RT PCR. RESULTS Compared to Pi 1 5 mmol?L -1 group,bovine smooth muscle cells (BSMC) cultured in medium containing Pi 2 0 mmol?L -1 phosphate level increased calcium deposition 〔On day 6, (77 187?11 692) mg?g -1 Pro versus (25 768?1 750) mg?g -1 Pro, P