Objective To investigate the possible mechanism of glomerular injury in diabetes mellitus by determining whether epithelial-mesenchymal transition (EMT) is caused by high glucose in mice podocytes. Methods Using mice glomerular podocyte cell line as an in vitro system, podocytes were incubated with glucose(12.5 mmol/L, 25 mmol/L, 50 mmol/L) and mannitol (50 mmol/L) for 36 hours. Then the cells were collected and expression of alpha-smooth muscle actin(α-SMA), fibronectin (FN), CD2 associated protein (CD2AP) and Wilms' tumor 1 gene (WT-1) was detected by Western blot and indirect immunofluorescence staining. Results Under low glucose (5.6 mmol/L) and mannitol (50 mmol/L) condition, there were high expression of CD2AP and WT-1, and low expression of α-SMA and FN in mice podocytes. After 36 hours treatment with high glucose (12.5 mmol/L), the expression of α-SMA and FN in podocytes was significantly increased, and the expression of α-SMA and FN was further up-regulated with the increase of glucose dosage (25, 50 mmol/L). The indirect immunofluorescence staining revealed the similar result, and the percentage of positive α-SMA cells was also increased compared with low glucose and mannital group (P<0.05). Meanwhile, Western blot showed that high glucose could down-regulate the expressions of CD2AP and WT-1 in a dose-dependent manner. Conclusion EMT may be a potential pathway leading to podocyte dysfunction and glomerular injury under high glucose conditions.

ObjectiveTo investigate the influence of fluid balance and model of renal replacement therapy (RRT) on renal function and prognosis of patients suffering from septic acute kidney injury (AKI).Methods A retrospective cohort analysis of 117 septic AKI patients who had undergone RRT between January 2009 and December 2014 was performed in the Second Affiliated Hospital of Nanjing Medical University. The patients were divided into positive fluid balance group (n = 52) and negative fluid balance group (n = 65) according to the total amount of fluid calculated from the difference between fluid administered and fluid lost during the first 1 week of RRT. The incidence of renal recovery and death of the patients by 60 days as the endpoint events were taken to judge the prognosis of two groups. RRT strategies included continuous renal replacement therapy (CRRT) and intermittent renal replacement therapy (IRRT). Multiple factors including estimated glomerular filtration rate (eGFR), sequential organ failure assessment (SOFA) score, RRT model, the accumulation of fluid before initiation of RRT, and negative fluid balance during RRT were analyzed for outcome predictors by Cox proportional hazards model.Results There were no differences between two groups regarding clinical characteristics. The percentage of receiving CRRT in the negative fluid balance group was slightly higher than that of the positive fluid balance group (52.31% vs. 36.54%,χ2 = 2.899,P = 0.089). With Kaplan-Meier survival curves, it was shown that the patients of negative fluid balance group had a higher rate of recovery of renal function (χ2 = 4.803,P = 0.028) and significantly lower mortality rate (χ2 = 9.505, P = 0.002). The rate of recovery of renal function by 60 days was higher in the negative fluid balance group than that in the positive fluid balance group (47.69% vs. 28.85%,χ2 = 3.991,P = 0.046), while the mortality rate was significantly lowered in the negative fluid balance group compared with that of the positive fluid balance group (40.00% vs. 67.31%,χ2 = 4.378,P = 0.036). Cox multivariate regression was used for excluding confounding factors. After adjusting for the clinically relevant variables, RRT negative fluid balance was significantly associated with recovery of renal function [hazard ratios (HR) = 2.440, 95% confidence intervals (95%CI) = 1.089-5.464,P = 0.030] and mortality (HR = 0.443, 95%CI = 0.238-0.822,P = 0.010]. Higher eGFR before RRT and CRRT were independent favorable factors for recovery of renal function (HR= 1.014, 95%CI = 1.003-1.026,P = 0.012;HR = 3.138, 95%CI = 1.765-7.461,P = 0.002), and higher SOFA score was associated with a significantly higher risk of death (HR = 1.115, 95%CI = 1.057-1.177, P< 0.001).ConclusionsOnce the patients with septic AKI showed the signs of fluid overload, timely RRT and effective removal of excessive liquid may reverse the adverse prognosis. RRT with negative fluid balance is beneficial for the recovery of renal function, and reduce the mortality in patients with septic AKI, and CRRT model is a good choice.

Objective:To explore the clinical characteristics of Haff disease in our hospital,be familiar with the disease and provide some experience in diagnosis and treatment.Methods:Inpatients with Haff disease in our Kidney disease center between July and August 2016 were retrospectively analyzed.The clinical data of patients was collected and statistically analyzed.Results:There were 66 patients (28 of male and 38 of female) in total with Haff disease between July and August 2016 in our ward.The average onset age was 35.5 years old (18-76 years old) and the average latency period was 5.6 hours (1-24 hours).All the patients had crayfish before the onset of the disease.The initial symptoms included muscle pain and concentrated brown urine (19 cases,28.8％).Laboratory tests suggested that transaminase and myotropin were increased significantly (alanine aminotransferase,aspartate aminotransferase,lactate dehydrogenase,creatine kinase,myoglobin and creatine kinase isoenzyme).A trace of blood (45％) and protein (16.7％) was present in some patients' urine test.After treatment,muscle pain was alleviated and urine color turned to clear in all patients.All the transaminase and muscle enzymes were significantly decreased (P＜0.05).No blood or protein was detected in the urine test.Conclusions:The epidemiology of Haffdisease in summer at Nanjing was related to the intake of crayfish.The latency period was short and the initial symptoms were mostly muscle soreness,with or without myoglobinuria.Laboratory tests showed serum creatine phosphokinase,myoglobin,creatine kinase isoenzymes were increased significantly.The treatment period was short with good prognosis.Generally no sequelae was observed.

Objective:To evaluate the clinical application value of iohexol plasma clearance assay in assessing glomerular filtration rate (GFR) in patients with chronic kidney disease (CKD), and identify alternative methods of 99mTc-diethylene triamine pentaacetic acid renal kinetic imaging (Gates) method for measuring GFR ( 99mTc-mGFR). Methods:It was a cross-sectional study. The CKD patients hospitalized in the Department of Nephrology at the Second Affiliated Hospital of Nanjing Medical University between October 2022 and September 2023 were enrolled. Iohexol plasma clearance was determined by collecting blood samples at 2 and 4 hours after intravenous administration of 5 ml iohexol, and high performance liquid chromatography was used to measure the plasma concentration of iohexol. Br?chner-Mortensen and Jacobsson formulas were used to calculate the double and single plasma iohexol clearance, respectively. CKD-epidemiology collaboration equation based on serum creatinine concentration was used to calculate the estimated GFR (eGFR). Pearson's coefficient was performed to analyze the correlation of iohexol dual plasma clearance assay-measured GFR (iohexol-DS-mGFR), iohexol single plasma 4 hours clearance assay-measured GFR (iohexol-SS 4h-mGFR), iohexol single plasma 2 hours clearance assay-measured GFR (iohexol-SS 2h-mGFR) and eGFR with 99mTc-mGFR as the gold standard. Bland-Altman analysis, 95% limits of agreement, and intra-correlation coefficient were used to compare the diagnostic concordance of Iohexol-DS-mGFR, Iohexol-SS 4h-mGFR, Iohexol-SS 2h-mGFR and eGFR with 99mTc-mGFR. Results:The study enrolled 64 CKD patients, aged (58.91±13.08) years old, comprising of 38 males and 26 females. The distribution of patients across CKD stages based on 99Tc-mGFR was as follows: 12 patients (18.8%) in stage 1, 14 patients (21.9%) in stage 2, 26 patients (40.6%) in stage 3, 10 patients (15.6%) in stage 4, and 2 patients (3.1%) in stage 5. The Pearson correlation analysis revealed that the correlation coefficients of iohexol-DS-mGFR, iohexol-SS 4h-mGFR, iohexol-SS 2h-mGFR and eGFR with 99mTc-mGFR were 0.925, 0.867, 0.820 and 0.894 (all P<0.001), respectively. The median deviation of absolute value of iohexol-DS-mGFR, iohexol-SS 4h-mGFR, iohexol-SS 2h-mGFR and eGFR in the total study population were 6.66, 9.63, 11.47 and 9.59 ml·min -1·(1.73 m 2) -1, respectively. The proportions of iohexol-DS-mGFR, iohexol-SS 4h-mGFR, iohexol-SS 2h-mGFR and eGFR located in the gold-standard GFR ± 10% interval (P10) were 35.9%, 29.7%, 26.6% and 29.7%, respectively, and the proportions located in the gold-standard GFR ± 30% interval ( P30 ) were 87.5%, 68.8%, 60.9% and 73.4%, respectively. Conclusions:Iohexol dual plasma clearance is well correlated and consistent with 99mTc-mGFR. It is a safe and easy alternative to isotope 99mTc-mGFR for clinical use in determining GFR.

Objective:To investigate the evaluation value of one-stop whole-brain dynamic volume CT angiography and CT perfusion imaging (CTA-CTP) in the cynomolgus monkeys models of middle cerebral artery occlusion (MCAO).Methods:Ten adult cynomolgus monkeys were selected and examined by head and neck CTA-CTP and craniocerebral MRI to rule out craniocerebral space-occupying lesions or cerebrovascular malformation. Under guidance of digital substraction angiography (DSA), the right femoral artery was dissected and monkey autologous thrombosis was injected into the right middle cerebral artery (MCA) through microcatheter to prepare MCAO models. Whole brain DSA was performed intraoperatively to observe whether the model was successfully prepared, and head and neck CTA-CTP was performed 24 h and 7 d after modeling to determine the locations and brain blood flow changes of ischemic lesions. The monkeys were sacrificed 8 d after modeling, and the brain tissues were stained with 2,3,5-triphenyltetrazolium chloride (TTC).Results:Among the 10 cynomolgus monkeys, one was excluded because of preoperative cerebrovascular malformation, and one died of cerebral hernia caused by cerebral hemorrhage during the experiment. The remaining 8 MCAO models were successfully prepared. Intraoperative DSA orthography showed unclear M1 segment and distal branch of MCA. Brain CT scan 24 h and 7 d after modeling showed obvious cerebral ischemic lesions in the right MCA blood supply area, and the infarct extent 7 d after surgery was more obvious than that 24 h after surgery. CTA examination showed obvious blood flow interruption imaging in the in M1 segment of MCA on the right side, the distal vessels were not clearly displayed and the distal branches of the infarct side 7 d after surgery were obvious decreased as compared with those 24 h after surgery. CTP scan showed that the cerebral blood volume of the right cerebrum was obviously reduced as compared with that of the left cerebrum, which was consistent with the blood supply area of MCA; and the infarct cores and penumbra areas 7 d after surgery were obvious increased as compared with those 24 h after surgery. TTC staining showed that the ischemic lesions of the brain tissue on the slices were gray and involved multiple layers, and the range was roughly consistent with the infarction sites shown by DSA and CT imaging.Conclusion:One-stop whole brain dynamic volume CTA-CTP has good evaluation value in imaging findings in MCAO animal models.