Objective    To investigate the correlation of M-type phospholipase A2 receptor (PLA2R) genetic polymorphism in two single nucleotide polymorphisms (SNPs) with idiopathic membranous nephropathy (IMN) of Chinese Han population in Northeast China.    Methods    A total of 327 individuals were enrolled in the study including 95 adult patients with biopsy-proved IMN (IMN group) followed up for (25.4±11.6) months and 232 healthy people identified by healthy examination in China-Japan Union Hospital of Jilin University (HC group). Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the genotype and allele frequency of rs35771982 and rs3828323 site in PLA2R gene. The χ² test was performed to compare the distribution difference of allelic frequency and genotype frequency of the two sites in PLA2R gene between two groups. Unconditional Logistic regression analysis was used to determine the risk factor of IMN.    Results    IMN and HC group were matched in male predominance and body mass index (BMI). Patients with IMN were older than the healthy controls and had higher Scr, serum total cholesterol (TC), 24-hour urine protein level and lower serum albumin (Alb) level, lower estimated glomerular filtration rate (eGFR) than the healthy controls (all P＜0.01). The CC genotype frequency and the C allele frequency at SNP rs35771982 site of PLA2R gene in IMN group were significantly higher than those in HC group ( χ²＝13.658, P＝0.001; χ²＝15.315, P＝9.10×10^-5), whereas there was no distribution difference of genotype and allele frequency at rs3828323 site between two groups (χ²＝2.844, P＝0.241; χ²＝2.959, P＝0.085). The CC genotype at rs35771982 site in patients with IMN was not related to age, gender, BMI, blood pressure and several laboratory indexes such as Alb, TC, Scr, eGFR and 24-hour urine protein level (all P＞0.05). Unconditional Logistic regression analysis revealed that the genotype at rs35771982, age, TC, Scr and eGFR were correlated with IMN occurrence. The CC genotype at rs35771982 was the risk factor of IMN (OR＝4.408, 95%CI 1.488-13.058).    Conclusions    The CC genotype and C allele at rs35771982 site in PLA2R may be associated with the susceptibility to IMN, whereas the correlation between gene polymorphism at rs3828323 site and IMN is not demonstrated. The CC genotype at rs35771982 is the independent risk factor of IMN in Chinese Han population in Northeast China.
 

Objective    To investigate the incidence, primary disease and vascular access of the initial hemodialysis patients in Shanxi province during 2010-2011.    Methods    A total of 3434 chronic renal failure (CRF) patients starting their first-time hemodialysis in Shanxi province during 2010-2011 were surveyed. Their data were registered in Chinese national renal data system. All related data were collected from Chinese national renal data system.    Results    In Shanxi province, 1514 cases began hemodialysis in 2010 and the annual incidence was 46.62 per million people (pmp), 1920 cases began hemodialysis in 2011 and the annual incidence was 53.44 pmp. The most common causes of CRF in these hemodialysis patients were glomerulonephritis (62.4%), diabetic  nephropathy (22.0%), and hypertensive nephrosclerosis (8.1%). The most popular vascular access in CRF patients at the beginning of hemodialysis was temporary central venous catheter (48.9%), then arteriovenous fistula (40.9%).    Conclusions    In Shanxi province, maintenance hemodialysis patients are increasing and there are more male patients. Main causes of ESRD patients on hemodialysis are chronic glomerulonephritis, diabetic nephropathy and hypertensive nephrosclerosis. The major vascular access of CRF patients at the beginning of hemodialysis is temporary central venous catheter, which   indicates that delayed hemodialysis is still a glaring problem in Shanxi province. 
   

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    Non-dipper blood pressure pattern is very common in CKD patients. Nighttime pressure is closely associated to renal damage and cardiovascular injuries. 
   

Objective    To analyze the efficacy and safety of total parathyroidectomy (PTX) with forearm autograft in uremic patients with secondary hyperparathyroidism (SHPT).    Methods    One hundred and eighteen cases undergoing PTX with forearm autograft in our hospital from 2001-2010 were included in this study. Their preoperative and postoperative serum intact parathyroid hormone (iPTH), biochemistry tests (total calcium，inorganic phosphate and alkaline phosphate) were collected and postoperative symptom relief, complications and recurrence were investigated.    Results    Of all the 118 cases, 32 underwent endoscopic surgery and 86 open surgery. The surgery was performed successfully in 110 cases (93.2%) and one case died in perioperative period. Thyroid carcinoma was diagnosed during surgery in 2 cases and radical operation was performed at the same time. Temporary injury of recurrent laryngeal nerve was found in nine cases (7.6%). Postoperative hypocalcemia was frequently seen in 108 cases (91.5%) and it was effectively controlled by postoperative calcium administration. After operation, bone pain and itching were alleviated, and weakness, anemia and malnutrition status were improved in all the cases who received successful surgery. The postoperative levels of serum iPTH (P＜0.01), calcium (P＜0.01), phosphorus (P＜0.01) and calcium×phosphorus (P＜0.01) were decreased significantly than those in preoperative period. A long-term follow-up of over 3 years was carried out in 21 cases. Six cases recurred, among them, 4 cases relieved after removal of autografted parethroid tissue, and another two cases received the second operation. The longest follow-up period lasted for 9 years in two cases without recurrence.    Conclusions    PTX with forearm autograft is safe and effective in the treatment for uremic patients with SHPT. No severe complication is found during the long-term follow-up period.

Objective     To evaluate the values of urinary liver-fatty acid binding protein      (uL-FABP) and urinary neutrophil gelatinase-associated lipocalin (uNGAL) in diagnosis of acute kidney injury (AKI) caused by obstructive nephropathy and in the prediction of renal prognosis.    Methods     Clinical data of 30 patients with obstructive nephropathy were collected prospectively. uL-FABP and uNGAL were measured by ELISA at various time points. Risk factors of the renal outcome were evaluated. The patients were followed up for at least one year.    Results    Patients with AKI had higher levels of uL-FABP and uNGAL compared to those without AKI [700.00(154.62-1216.14) μg/g•Cr vs 26.90 (16.77-41.38) μg/g•Cr; 1266.69 (671.57-3396.07) μg/g•Cr vs 179.12 (90.98-215.16) μg/g•Cr, all P＜0.01]. Positive correlations of uL-FABP and uNGAL with serum creatinine were found (r＝0.552, 0.553, all P＜0.01). The AUCs of uL-FABP and uNGAL to detect AKI were 0.925 and 0.900. Patients with non complete renal recovery had higher levels of uL-FABP before operation and 72-hour after operation compared to those with complete renal recovery (all P＜0.01). Before operation, the AUC of uL-FABP to detect renal prognosis was 0.948, sensitivity was 85.7% and specificity was 90.9%. 72-hour after operation, the AUC of uL-FABP to detect renal prognosis was 0.935, sensitivity was 85.7% and specificity was 90.9%. Kaplan-Meier analysis revealed that uL-FABP before operation over 366.57 μg/g•Cr or uL-FABP 72-hour after operation over 223.60 μg/g•Cr were closely related to the poor progression of renal function.    Conclusions    uL-FABP and uL-NGAL have good accuracy in detecting AKI. The level of uL-FABP before operation and 72-hour after operation is helpful to predict the renal outcome of obstructive nephropathy.
 

Objective    To evaluate the effects of AngⅡ on the expression of IQ domain GTPase-activating protein1 (IQGAP1) and apoptosis of glomerular cells, and to explore the role of IQGAP1 in AngⅡ-induced apoptosis of glomerular cells.    Methods    Thirty-six male Wistar rats were randomly assigned to receive either saline or AngⅡ by osmotic mini-pump, or be used as normal control. The systolic blood pressure and proteinuria were measured at day 7, 14, 21, 28. After the animals were sacrificed at day 14, 28 respectively, the kidneys were collected. Renal pathological change, glomerular cell apoptosis were observed. The expression of glomerular IQGAP1 was assessed by immunohistochemistry, immunofluorescence and Western blotting. The activation of caspase-3 and phosphorylation of extracellular regulated protein kinases 1 and 2 (ERK1/2) were determined by Western blotting.    Results    AngⅡ-infused rats developed significant hypertension and marked proteinuria. Mild glomerular mesangial cell proliferation and mesangial matrix increase were also observed in AngⅡ-infused rats. The number of apoptotic glomerular cells in AngⅡ-infused rats was significantly more than that in normal control (P＜0.05). The expression of IQGAP1 in glomeruli distributed linearly along the capillary loops.  AngⅡ infusion up-regulated the expression of glomerular IQGAP1, which had an significantly positive correlation with activation of caspase-3(r＝0.689, P＜0.05) and phosphorylation of ERK1/2 (r＝0.658, P＜0.05).    Conclusion    The enhanced expression of IQGAP1 may be involved in AngⅡ-induced glomerular cells apoptosis via activation of ERK1/2 signaling pathway.
 

Objective    To investigate the effect of benazepril on intergrin-linked kinase (ILK) and α-smooth muscle actin (α-SMA) expression in glomerular mesangial cells induced by high-glucose.    Methods    The mesangial cells from SD rat (HBZY-1) were cultured conventionally and randomly divided into four groups: normal glucose (D-glucose 5.5 mmol/L, group NG), mannitol-treated  group (mannitol 20 mmol/L, group MG), high glucose (D-glucose 30 mmol/L, group HG), Benazepril-treated high glucose group (D-glucose 30 mmol/L＋Benazepril 10 μmol/L, group ACEI). Cells from NG, MG, HG, ACEI gronps were harvested after 3, 6, 12, 24, 48 and 72 hours of treatment respectively. The mRNA expressions of ILK and α-SMA were detected by RT-PCR. The protein levels of ILK and          α-SMA were detected by Western blotting and immunofluorescence.    Results    The expressions of ILK mRNA and protein in HG group were significantly increased compared with those in NG group (all P＜0.05). The increased expressions of ILK and α-SMA in HG group were time-dependent and the expression reached the peak at 48 h (ILK, P＜0.05) or 72 h (α-SMA, P＜0.01). The expressions of ILK and α-SMA in ACEI group were lower than those in HG group (all P＜0.01), but failed to rescue to the same level as those in NG. There was no significant differences of ILK expressions between MG group and NG group at the same time point (P＞0.05). The expressions of α-SMA mRNA and protein in MG were higher than that in NG (P＜0.05), which suggest that high osmotic pressure could cause the increasing of α-SMA.    Conclusions    Benazepril can decrease the expressions of ILK and α-SMA to inhibit the process of fibrosis in DN and mediate the phenotypic transformation of glomerular mesangial cells. The phenotypic transformation of glomerular mesangial cells in glucose may also depend on high osmotic pressure in DN.
  

Objective    To observe the cell proliferation and the protein expression of STAT1,phosphorylation of STAT1 (p-STAT1), STAT3, p-STAT3 and transforming growth factor β1 (TGF-β1) in human glomerulur mesangial cells (HMCs) induced by high glucose after STAT1-siRNA transfection.   Methods    Three STAT1-siRNA sequences were designed and synthetized. HMCs in 6-well plate were transiently transfected with STAT1-siRNA using Lipofectamine 2000. After transfection for 48 h or 72 h, STAT1 mRNA and protein expression were detected by real-time PCR and Western blotting, respectively, to choose the effective sequence in later experiments. After transfection for 24 h and stimulated with 25 mmol/L glucose for 24 h, 48 h, 72 h, cell proliferation was measured by MTT assays, the protein expressions of STAT1, p-STAT1, STAT3 and p-STAT3 were detected by Western blotting, the expression of TGF-β1 was detected by ELISA in each group.    Results    High glucose could stimulate HMCs proliferation. The protein expressions of p-STAT1, p-STAT3 and TGF-β1 were increased in the group stimulated by high glucose (P＜0.05). The protein expressions of p-STAT3 and TGF-β1 were further increased in HMCs induced by high glucose after STAT1-siRNA transfection (P＜0.05).    Conclusions    Under high glucose conditions, JAK-STAT signal transduction pathway of HMCs can be activated, then it is far greater when HMCs are induced by high glucose after STAT1-siRNA transfection. The secretion of TGF-β1 is increased in HMCs under the state of high glucose, and it is further increased after STAT1-siRNA transfection, which is related to the kidney fibrosis.
 

Objective    To investingate the effect of low-density lipoprotein (LDL) on epithelial-mesenchymal transition and extracellular matrix (ECM) accumulation in human peritoneal mesothelial cells (HPMCs).    Methods    (1)HPMCs were randomly divided into control group, LDL group (100 mg/L) and LDL (100 mg/L)＋lactoferrin (100 mg/L, LDL receptor blocking agent) group. After co-cultured for 24 h, the expression of LDL receptor in HPMCs was examined by immunofluorescence staining, and the LDL uptake by HPMCs was observed with oil red O staining. (2)HPMCs were cultured with different concentrations of LDL (0, 25, 50, 100 mg/L). After co-cultured for 24 h, the change of cell morphology was observed by inverted phase contrast microscope, and the expression of α-smooth muscle actin (α-SMA) was examined by immunofluorescence. (3) HPMCs were randomly divided into control group (5.6 mmol/L glucose), mannitol group (M, 2.18% mannitol), low glucose group (LG, 30 mmol/L), high glucose group (HG, 120 mmol/L) and HG＋LDL group (120 mmol/L glucose＋100 mg/L LDL). Co-cultured for 48 h, the mRNA expression of α-SMA, E-cadherin and type 1 plasminogen activator inhibitor (PAI-1) was detected by real-time quantitative PCR, the protein expression of α-SMA was detected by Western blotting, the content of type I collagen (ColⅠ) and PAI-1 in supernatant was detected by ELISA.    Results    (1) After co-cultured with LDL for 24 h, the expressin of LDL receptor was found on the cell membrane of HPMCs. Oil red staining showed that LDL could be uptaken into the cells and abolished by LDL receptor blocker. (2) HPMCs tended to be loosely intercellular connected to each ofher, and prsesnted significant formation of fibroblast?like spindle morphology. The cytoplasm immunofluorescence intensity of α-SMA gradually increased with the increase of LDL concentration. Compared to the control group, the expressions of α-SMA mRNA and protein were significantly increased, and the expression of E-cadherin mRNA was decreased in HG＋LDL group(all P＜0.05). But the expressions of the parameters above-mentioned were not significant different between HG group and HG＋LDL group or between HG group and control group. (3) Compared with HG group or control group, the concentrations of ColⅠ[(19.27±0.17) μg/L vs (14.09±0.30) μg/L or (14.81±0.91) μg/L, all P＜0.05] and PAI-1 [(498.24±76.91) ng/L vs (342.19±30.43) ng/L or (220.39±33.82) ng/L, all P＜0.05] in supernatant of HPMCs were significantly up-regulated in HG＋LDL group, meanwhile the expression of PAI-1 mRNA was significantly higer than that in control group (P＝0.022）.    Conclusions    HPMCs uptake LDL into cells via LDL receptors. LDL can induce HPMCs transdifferentiation in the condition of high glucose, increase the secretion of ColⅠ, inhibit the degradation of ECM through up-regulating the expression of PAI-1, and lead to ECM accumulation. 

Objective    To investigate the effect of hepatitis B virus X (HBX) gene on apoptosis and immune molecules of human proximal renal tubular epithelial cell line (HK-2).    Methods    The eukaryotic vector pcDNA3.1-myc-HBX containing HBX gene was transiently transfected into HK-2 cells by lipofectamine mediation. Untransfected HK-2 cells and those transfected with empty vector were used as controls. The TLR4 expression was detected by real-time PCR and Western blotting. The apoptosis of cells and expression of MHC-Ⅱ and CD40 were detected by flow cytometry, and the contents of IL-4 and IFN-γ in the supernatant were detected by ELISA.    Results     Compared with control groups, the number of apoptotic cells was significantly increased in the HBX transfection group (P＜0.05), and the expressions of TLR4, MHC-Ⅱ and CD40 were also significantly increased in the HBX transfection group (all P＜0.05). IFN-γ level in the supernatant of HBX transfection group was higher (P＜0.05), but IL-4 level was lower as compared to control groups (all P＜0.05).    Conclusions    Over-expression of HBX gene may induce apoptosis of HK-2 cells and up-regulate the expression of immune molecules of renal tubular epithelial cells leading to injury of cells and dysfunction of immunomicroenviroment.