Objective    To analyze the prevalence, risk factors of kidney disease in type 2 diabetic patients with KDIGO classi?cation of chronic kidney disease, also to study cardiovascular and cerebrovascular diseases and death in these patients, so as to investigate the significance of the KDIGO classification system.    Methods    One thousand six hundred and forty-five type 2 diabetic patients who were in hospitalization from June 2008 to December 2012 were grouped according to the KDIGO classification of chronic kidney disease and the incidence of vascular disease was analyzed based on the classification. Clinical features were compared between patients with or without kidney disease. The risk factors of kidney disease and the death of diabetic patients were also investigated.    Results    There were 915 male and 730 female, aged a median (57.86±12.54) years with (6.35±6.30) years duration of diabetes mellitus among the 1645 cases, and 37.2% of patients had concomitant kidney disease. According to the classi?cation of CKD, patients in CKD group 3a, group 3b and CKD group 4-5 accounted for 5.7%, 3.5% and 7.6%, while 33.4% of patients had proteinuria, among which 19.5% with microalbuminuria, 13.5% with macroalbuminuria. On complications, patients with hypertension accounted for 49.5%, hyperlipidemia 67.7%, diabetic retinopathy 27.4%, cardiovascular and cerebrovascular diseases 18.5%(coronary artery disease 16.5%, cerebrovascular diseases 8.8%). Statistical difference was detected in the incidence of diabetic retinopathy, coronary artery disease and cerebrovascular diseases between CKD group 3a and 3b (P＜0.05). The duration of diabetes, concomitant hypertention especially with elevated systolic blood pressure, diabetic retinopathy and hyperuricemia were the independent risk factors for type 2 diabetic patients with kidney disease. Age, Scr, complicating cardiovascular and cerebrovascular diseases and advanced CKD stage were the independent risk factors for the death of type 2 diabetic patients with kidney disease.    Conclusion    KDIGO classification of chronic kidney disease enables better staging of kidney diseases in diabetic patients for management and prognosis. Diabetic patients have a higher prevalence of renal diseases and cardiovascular and cerebrovascular events than the general population. Early control of factors such as blood pressure and serum uric acid can delay the progression of kidney disease, and the predictive role of diabetic retinopathy should be emphasized.   

Objective    To investigate the expression of serum adiopocyte fatty acid binding protein(A-FABP) in chronic kidney disease (CKD) and the role that A-FABP plays in CKD with atherosclerosis.    Methods    A total of 138 patients with CKD and 20 health control volunteers (HC) were involved in this study. The levels of serum A-FABP, free fatty acid (FFA), interleukin- 6 (IL-6), monocyte chemotactic protein 1(MCP-1) were measured by enzyme-linked immunosorbent assay(ELISA). Inteima-media thickness of common carotid artery was measured by color doppler ultrasound. Results    According to the progression of glomerular filtration rate(GFR), the patients with CKD were divided into three groups: group CKD1-2[eGFR≥ 60 ml·min-1·(1.73 m2)-1], group CKD 3-4[60 ml·min-1·(1.73 m2)-1 > eGFR ≥ 15 ml·min-1·(1.73 m2)-1], group CKD5[eGFR ＜ 15 ml·min-1·(1.73 m2)-1].The levels of serum A-FABP were relatively higher in CKD than that in HC group(P＜0.05), and that in the group CKD5 were the most highest (P＜0.01). The level of serum FFA in group CKD 1-2 was relatively higher than that in group HC (P＜0.05), and FFA had a rising trend with decreased eGFR. The level of serum A-FABP was positively correlated with the levels of serum FFA (r＝0.825, P＜0.01), and also positively correlated with IL-6 (r＝0.569, P＜0.01), MCP-1(r＝0.657, P＜0.01) in CKD by Pearson correlation analysis. The levels of A-FABP in 56 patients of CKD with vascular atherosclerosis were significantly higher than that in 82 patients without vascular atherosclerosis (P＜0.01).    Conclusion    Serum A-FABP maybe play an important role in the progression of vascular atherosclerosis in CKD.  

Objective    To understand the blood pressure variability (BPV) and the influencing factors through ambulatory blood pressure monitoring during hemodialysis (HD) in the end-stage renal disease (ESRD) patients.    Method    Eighty-one ESRD patients on maintenancing HD for more than three months were enrolled into the study. The patients were with properly dry body weight. The blood pressure was monitored using dynamic blood pressure monitor around the HD. BPV was estimated with the coefficient of variation (CV) and standard deviation (SD) of the systolic blood pressure (SBP-CV, SBP-SD). Patients were divided into two groups according to the mean of SBP-CV: high SBPV group and low SBPV group. The possible influencing factors such as age, dialysis duration, ultrafiltration volume, ultrafiltration/body weight, therapy of antihypertensive, electrolyte, nutrition state, metabolic bone disease indexes, inflammatory state and serum lipid state were analyzed and compared between the two groups. And multivariate stepwise regression analysis was made between the SBP-CV, SBP-SD and the above observational parameters.    Results    The average SBP-CV of the 81 patients was (8.12±3.16)%, SBP-SD was (11.22±4.55) mm Hg. The proportion of hypertention and hypotention in high SBPV(SBP-CV≥8.12%) group (20.0%, 25.7%) was higher than that in the low SBPV(SBP-CV＜8.12%) group (8.7%, 6.5%)(P＝0.009). Serum high-sensitivity c-reactive protein (hs-CRP) and alkaline phosphatase (ALP) were higher in high SBPV group than that in the low SBPV group[(7.19±5.95) mg/L vs (3.35±2.78) mg/L, P＝0.001 and (180.31±96.32) U/L vs (98.00±41.19) U/L, P＝0.049]. Serum creatinine and potassium were higher in the low SBPV group than that in the high SBPV group   [(1015.83±276.20) μmol/L vs (893.63±216.61) μmol/L, P＝0.034 and (5.27±0.78) mmol/L vs (4.80±0.23) mmol/L, P＝0.005]. SBP-SD was positively correlated with hs-CRP (β＝0.499, P＜0.01), SBP-CV was positively correlated with hs-CRP and dialysis vintage (β＝0.464 and 0.211, P＜0.01 and P＜0.05) by the multivariate stepwise regression analysis.    Conclusions    The SBP-CV during HD is 8.12% in ESRD patients. Hypertention and hypotention are more often in the higher SBPV patients. SBPV is closely related to the serum hs-CRP.

Objective    To investigate the association between endothelial dysfunction and arterial stiffness in continuous ambulatory peritoneal dialysis （CAPD） patients.    Methods    Ninety-four stable CAPD patients from a single center were enrolled in this cross-sectional study. Ultrasound evaluation was conducted on brachial artery to estimate endothelial-dependent flow-mediated dilation (FMD). Automatice pulse wave velocity (PWV) measuring system was applied to examine the carotid-femoral PWV. Blood pressure and biochemical parameters were detected. Pearson's correlation and Stepwise multiple regression analysis were performed to explore the relationship between FMD and PWV.    Results    PWV was significantly higher in patients with diabetes as compared to those without diabetes[(13.25±1.66) m/s vs (11.24±1.92) m/s, P＜0.01]. Furthermore, PWV was positively correlated with age(r＝0.319, P＝0.002), SBP (r＝0.289, P＝0.005) and C-reactive protein (r＝0.211, P＝0.041), was negatively correlated with albumin (r＝-0.429, P＝0.001) and FMD (r＝-0.466, P＝0.001). In multivariate regression analysis, diabetes mellitus, albumin, FMD, age and SBP were independently associated with PWV after adjustment.    Conclusion    Endothelial dysfunction is associated with greater arterial stiffness in CAPD patients.

Objective    To investigate the changes of serum leptin levels and the influential factors in maintenance peritoneal dialysis patients.    Methods    Seventy-six peritoneal dialysis patients were chosen at the time before, and 3 months, 6 months, 12 months, 18 months and 24 months after they began the peritoneal dialysis therapy, to examine body mass index (BMI), triceps skinfold thickness (TSF), abdominal circumference, homeostasis model assessment of insulin resistance (HOMA-IR), the plasma lipid profile, and leptin in the same situation.    Results    For 24 months, these patients showed higher serum leptin level than the values before commencing peritoneal dialysis treatment (P＜0.01). The level of leptin was positively correlated with the BMI(r＝0.412, P＜0.01), TSF(r＝0.308, P＜0.01), abdominal circumference(r＝0.284, P＜0.01), HOMA-IR(r＝0.184, P＜0.01) and TG(r＝0.288, P＜0.01), negatively corelated with the high-density lipoprotein cholesterol(HDL-C)(r＝-0.285, P＜0.01). Multiple logistic regression analysis showed that BMI (β＝0.339, P＜0.01), TG(β＝0.157, P＜0.01) and HDL (β＝-0.126, P＜0.05)were significant predictive factors for the changes of serum leptin levels.    Conclusion    Leptin maybe involve in the occurrence and the development of cardiovascular events like other metabolic parameters in peritoneal dialysis therapy.

Objective    Plasma phospholipid metabolism analysis was employed to investigate plasma metabolic profile of acute renal graft rejection in order to find potential disease biomarkers and reveal its pathophysiological changes.    Methods    Selected 33 patients which did renal transplant in our hospital during 2009 to 2010, named preoperative group. 14 patients were diagnosed as acute graft rejection, and 19 patients were non-acute graft rejection. Then use ultra performance liquid chromatography (UPLC) coupled with quadrupole time of flight mass spectrometry (qTOF-MS) to establish the metabolic fingerprint in those patients. The date was preprocessed with software Markerlynx, and analyzed with partial least squares discriminant analysis    (PLS-DA), and data from conventional laboratory techniques were used for assessing renal function.    Results    In the plasma of renal transplant patients, seventeen biomarkers were discovered: creatinine, four sphingomyelins (SM), nine phosphatidylcholines (PC), three lysophosphatidylcholine (LPC). While the creatinine was increased, the others were reduced obviously.    Conclutions    UPLC-qTOF-MS based metabonomics can analyze the plasma phospholipid metabolism in patients with renal transplantation, reveal the law of phospholipids metabolic changes in the process of acute rejection. The levels of plasma phosphatidylcholine's esterlysis and fatty acids' β-oxidation are increased. The immunosuppressive therapy can reduce the activity of phospholipase, inhibit the level of plasma phosphatidylcholine's esterlysis and fatty acids' β-oxidation, which restraines the development of acute rejection after renal transplantation. Therefore it may be a promising new technology in diagnosing acute renal graft rejection after renal transplantation.   

Objective    To observe the regulation of Toll-like receptor 4 (TLR4) signal and the release of inflammation factors after angiotensin II (AngⅡ) stimulation in rat mesangial cells under high glucose condition, revealing the innate immune-related mechanism of injury by AngⅡ on mesangial cells under high glucose.    Methods    After synchronization, cells incubated with AngⅡ(10-7 mmo/L) and/or high glucose (25 mmol/L) were used as the stimulation group, cells without stimulation were as normal control (5.6 mmol/L glucose).    To determine the role of TLR4 and the adaptor myeloid differentiation factor 88 (MyD88), equal number of HBZY-1 cells were added with 10-5 mmol/L irbesartan and/or TLR4 blocker (10 mg/L) for 1 h and then incubated with AngⅡ (10-7 mmo/L) and/or high glucose (25 mmol/L) for 12 h or 24 h respectively. Real-time PCR was used to analyze TLR4 mRNA and MyD88 mRNA expression after 12 h. Immunofluorescence was used to observe TLR4 protein expression after 24 h; Western blotting was used to observe TLR4, MyD88 and nuclear factor κB (NF-κB) protein; ELISA was used to detect the concentration of MCP-1, IL-6 in cell supernatant respectively.    Results    Compared with normal control group, TLR4 mRNA and MyD88 mRNA were highly expressed in high glucose or AngⅡ-induced HBZY-1 cells (P＜0.01), TLR4, MyD88 and       NF-κB protein as well as MCP-1, IL-6 were also up-regulated significantly (P＜0.01). Compared with high glucose or AngⅡ group, MyD88 and NF-κB protein as well as MCP-1, IL-6 were further            up-regulated markedly in AngⅡ and high glucose costimulated group (P＜0.01)．In HBZY-1 cells that were preincubated with irbesartan and/or TLR4 blocker, TLR4 and MyD88 protein expression were obviously inhibited, IL-6 and MCP-1 production were also decreased remarkably compared with high glucose and/or AngⅡ group (P＜0.01).    Conclusions    High glucose and AngⅡ stimulate the release of proinflammatory factors in rat glomerular mesangial cells via TLR4-MyD88 pathway. This process is inhibited by irbesartan or TLR4 blocker via modulation of the signal. AngⅡ has the positive-regulation potential on the release of inflammation factors via TLR4 signal in rat mesangial cells under high glucose condition.

Objective    To assess the characteristics of different doses of cisplatin-induced acute kidney injury, further to understand mitochondrial dysfunction and its role in acute kidney injury (AKI).    Methods    Male C57BL/6J mice were first randomly divided into two groups: control group (n＝6) and AKI group (n＝12). Then, AKI group was subsequently divided into other two groups according to different dose of cisplatin (10 mg/kg or 20 mg/kg). AKI group received intraperitoneal injection of cisplatin. All mice were sacrificed after 72 h of injection. Renal biochemical function, renal pathological changes, renal injury markers, kidney mitochondrial function and structural changes were observed.    Results    (1) After 72 hours of injection, the AKI group performed significant kidney injury changes compared to control group, thereinto 20 mg/kg group was more serious than 10 mg/kg group. With the cisplatin dose increasing, renal function markers such as serum creatinine, urine protein gradually increased. (2)Kidney biopsy showed tubular structural damage, the formation of protein casts, kidney injury molecule-1 (KIM-1) gradually increased(P＜0.05). (3)Electron microscopy found tubular mitochondrial structural damage, mtDNA copy number decreased, the level of peroxisome proliferator-activated receptor -gamma coactivator-1alpha (PGC-1α), ATP synthase β decreased(P＜0.05), and Western blotting manifested cytochrome C was released from mitochondria to the cytoplasm. These data all exhibited significant difference between different groups(P＜0.05).    Conclusions    Cisplatin induces acute kidney injury in dose-dependent manner. Mitochondrial dysfunction participates in kidney injury, and is also related to the kidney pathological damage.

Objective    To investigate the effects of the rat serum with chronic renal failure(CRF) on ubiquitin-proteasome pathway, histone acetyltransferase p300 and activation of activating transcription factor 4(ATF4) of rat arterial vascular smooth muscle cells(VSMCs) cultured in vitro, and explore the possible mechanism.    Methods    To establish the rat model of CRF by 5/6 nephrectomy, VSMCs were incubated in the media with the 10% of CRF serum or control serum in vitro. The mRNA expressions of ubiquitin(Ub), ubiquitin activating enzyme(E1), ubiquitin ligases enzymes (β-transducin repeat containing protein 1, β-TrCP1), p300 and ATF4 in the rat VSMCs were examined by using real-time PCR. Expressions of E1, β-TrCP1, p300 and ATF4 proteins in response to the CRF serum in VSMCs were determined by Western blotting analysis. The enzyme activities of 20S proteasomes in the total protein were examined by using three special fluorogenic peptide substrates.    Results    The CRF serum significantly promoted the mRNA expressions of Ub, E1, β-TrCP1, p300 and ATF4 in VSMCs in a time dependent manner. Compared with that in control serum group, the mRNA levels of Ub, E1, β-TrCP1, p300 and ATF4 in CRF serum group increased significantly (P＜0.01). The CRF serum also increased the protein expressions of E1, β-TrCP1 and p300 in a time dependent manner. The expression of ATF4 was decreased, but the difference was not significant (P＞0.05). Compared with that in control serum group, the protein expressions of E1, β-TrCP1, p300 and ATF4 in CRF serum group increased significantly (P＜0.01). The activities of 20S proteasomes in the CRF serum group were significantly increased in a time dependent manner. Compared with that in control serum group, the activities of 20S proteasomes in the CRF serum group increased significantly (P＜0.01).    Conclusions    The serum of CRF rat can effectively active the ubiquitin-proteasome pathway, but ATF4 ubiquitinylated degradation is blocked. The latter may be associated with increased expression of p300.