Objective    To determine the relationship between changes of blood pressure (BP) during dialysis and mortality in maintenance hemodialysis (MHD) patients.    Methods    A total of 364 cases of MHD patients were collected prospectively and the relationship between changes of blood pressure during dialysis and mortality was assessed.    Results    The patients' age was (63.07±13.93) years. Over a follow-up of (54.86±19.84) months, a total of 85 (23.4%) all-cause and 46(12.6%) cardiovascular deaths occurred. Post-dialytic drops in systolic BP between 7.08 mmHg and 14.25 mmHg were associated with lower all-cause and cardiovascular mortality [OR=0.324 and 0.335, 95%CI (0.152, 0.692) and (0.123, 0.911), P=0.004 and 0.032, respectively]. Kaplan-Meier analysis showed that post-dialytic increase in systolic BP more than 0.25 mmHg was associated with higher all-cause and cardiovascular mortality (P=0.001, 0.044, respectively). Multivariate logistic regression analysis showed that post-dialytic increase in systolic BP more than 0.25 mmHg, hemoglobin, Kt/V were independent risk factors for all-cause mortality.    Conclusions    Post-dialytic increase in systolic BP more than 0.25 mmHg in MHD patients suggests higher mortality. Significant increased systolic BP after hemodialysis, hemoglobin level and Kt/V were independent risk factors for all-cause mortality.

Objective    To explore the correlation of Th17/Treg ratio and related cytokines with clinical and pathological activity in patients with lupus nephritis (LN).    Methods    The patients with lupus nephritis were enrolled into this study from June 2011 to Feb 2012. The demographic data, clinical activity and pathological index were recorded and analyzed in details. The frequency of Th17 and Treg in peripheral blood CD4+ T lymphocytes was tested by flow cytometry and the ratio of Th17 to Treg was calculated. The levels of such cytokines as IL-6, IL-10, IL-17, IL-23, TGF-β1 were detected by ELISA.    Results    A total of 60 patients with LN were enrolled in this study. Of them, 90% were female, 10% were male, the average age was (36.39±14.23) (14-75) years old, the duration was 1.00(0.33-9.25) years, the average SLEDAI was (9.50±4.61). Compared with control group, the ratio of Th17 to Treg in LN group was significantly higher [(0.95±0.67) vs (0.29±0.13), P＜0.05]; further the Th17/Treg ratio in active LN group was much higher [(1.33±0.71) vs (0.57±0.33), P＜0.01] compared with inactive LN group. The ration of Th17 to Treg was positively correlated with SLEDAI and AI (r＝0.650, P＜0.01; r＝0.675, P＜0.01). The level of serum IL-10 was negatively correlated with SLEDAI and AI(r＝-0.567, P＜0.01; r＝-0.422, P＜0.01). The level of serum IL-17 was positively correlated with SLEDAI and AI (r＝0.559, P＜0.01; r＝0.479, P＜0.01). The level of IL-23 was positively correlated with SLEDAI and AI (r＝0.339, P＜0.05; r＝0.350, P＜0.05). The levels of serum anti-dsDNA and C3 were not correlated with SLEDAI and AI in patients with LN (P＜0.01). As the diagnostic indicator of active LN(SLEDAI≥10), the AUCs of Th17/Treg, IL-17, IL-23 were 0.92 (0.83-1.00), 0.91 (0.81-1.00) and 0.67(0.51-0.83), respectively. As the diagnostic marker of AI＞8，the AUCs of Th17/Treg and IL-17 were 0.87(0.76-0.99) and 0.84 (0.69-0.99).    Conclusions    The ratio of Th17 to Treg and the level of serum IL-17 are good indicator for evaluating both clinical and pathological activity in patients with LN. The levels of IL-10 and IL-23 show better correlation with SLEDAI and AI than the traditional indicator anti-dsDNA and C3.

Objective    To investigate whether there is any difference in aortic stiffness among different hypertension subtypes in patients with chronic kidney disease.    Methods    Six hundred and twenty-six patients with chronic kidney disease were included in the present analysis. They were classified into four groups: normotension (n＝391) with systolic blood pressure (SBP) ＜140 mmHg and diastolic blood pressure (DBP) ＜90 mmHg; isolated systolic hypertension (ISH, n＝141) with SBP≥140 mmHg and DBP＜90 mmHg; isolated diastolic hypertension (IDH, n＝25) with SBP＜140 mmHg and DBP≥90 mmHg; systolic-diastolic hypertension (SDH, n＝69) with SBP≥140 mmHg and DBP≥90 mmHg. Aortic stiffness was assessed by pulse pressure and pulse wave velocity.     Results    The IDH group had lower mean age than the other groups(P＜0.01). The percentage of diabetes in the ISH group was higher than that in the other groups. The comparison of aortic stiffness showed that the ISH and SDH groups had higher aortic stiffness than the normotension and IDH groups (P＜0.01), but no significant difference in aortic stiffness was observed neither between the normotension and IDH groups nor between ISH and SDH groups. Conclusion   Aortic stiffness is significantly different among different hypertension subtypes, which may be an underlying cause for the different cardiovascular mortality among the hypertension subtypes.

Objective To investigate the possibility of urinary microRNA-29 (miR-29) as biomarker for diabetic nephropathy in patients with type 2 diabetes.    Methods    Sixty-one patients with type 2 diabetes were divided into 2 groups: diabetes with normoalbuminuria[n＝25, (58.88±11.75) years old] and diabetes with albuminuria[n＝36, (62.19±13.11) years old]. There were no significant differences in age and gender between groups. The contents of miR-29a, miR-29b and miR-29c in urine supernatant were determined by real-time quantitative PCR, and a synthetic cel-miR-39 was added to the urine as a spike-in control before miRNAs extraction. The laboratory parameters including urinary albumin excretion, serum creatinine, BUN, glycosylated hemoglobin, and blood lipids were collected, while retinopathy serves as non-invasive method to assess vascular fibrosis.    Results    There was no significant difference in glycosylated hemoglobin levels and duration of diabetes between two groups, while the diabetes with albuminuria group had lower estimated glomerular filtration rate (eGFR) (P＝0.001), had higher level of miR-29a, miR-29b and miR-29c in urine (P＝0.029, 0.032, 0.040) compared with diabetes normoalbuminuria group. Urinary albumin excretion rate significantly correlated with urinary miR-29a level (r＝0.284, P＝0.039) and miR-29b level (r＝0.275, P＝0.046), urinary miR-29b significantly correlated with BUN（r＝0.277, P＝0.031）in patients with type 2 diabetes. However, no correlation was found between miR-29a, miR-29b, miR-29c and other clinical parameters. Conclusion    Urinary miR-29a and miR-29b correlates with urinary albumin in patients with type 2 diabetes, and it needs further exploration and evaluation for urinary miR-29 to serve as potential biomarker for diabetic nephropathy in type 2 diabetes.

Objective    To investigate the association of single nucleotide polymorphisms (SNPs) of the mannan-binding lectin (MBL) gene with serum levels, development, progression and prognosis of severe lupus nephritis (LN).    Methods    A total of 107 severe lupus nephritis patients were enrolled in the study from January 2003 to October 2013. Integrated capillary electrophoresis was used to detect MBL gene polymorphism in peripheral blood DNA. ELISA was used to detect serum MBL concentration. Kaplan-Meier survival analysis was used to analyse the relationship of renal function, kidney prognosis with the gene polymorphism of rs11003125. Cox regression model analysis was used to identify possible risk factors of kidney prognosis.    Results    SNPs in rs11003125, rs7096206, rs7095891 and rs1800450 were found. The serum MBL concentration of patients with GG genotype in rs11003125 was higher than that with GC genotype, and both were higher than that with CC genotype (P＜0.01). Patients with SNP of rs11003125 had higher systolic blood pressure, diastolic blood pressure, mean arterial pressure, serum creatinine, urea nitrogen, 24 hours urinary protein, and lower glomerular filtration rate, shorter mean renal survival time (P＜0.05). Progressive severe LN patients had higher GC+CC (91.9% vs 75.7%, P＝0.041), CT+TT（32.4% vs 14.3%, P＝0.027） genotype frequencies at promoter rs11003125 and rs7095891, respectively, compared with that of non-progressive severe LN patients.    Conclusions    rs11003125, rs7096206 and rs1800450 polymorphisms of MBL gene are associated with lower serum MBL levels in severe LN patients. rs11003125 promoter polymorphisms of MBL gene may contribute to the onset severity, progression and prognosis of severe lupus nephritis.

Objective    To determine whether mutation of Hepatitis B virus (HBV) X gene is associated with hepatitis B virus-associated glomerulonephritis (HBV-GN).    Methods    The venous blood was collected from 50 patients with HBV-GN and 60 patients with asymptomatic HBV carriers (control group). Serum HBV DNA was extracted to determine the serum titer of HBV-DNA and then polymerase chain reaction (PCR) was used to detect the HBV X gene mutation.    Results    (1)There were not statistical significance between age and gender in HBV-GN group and control group (P＞0.05). There were not statistical significance of serum replication level of HBV DNA in HBV-GN with X gene mutation and control group (P＞0.05). Urine protein excretion in HBV-GN group with or without X gene mutation was found with statistical significance (P＜0.05). (2)Nucleotide mutations [84%(42/50)] resulted in amino acid substitution in HBV-GN. Nucleotide mutations changed in trans-function control region of X gene, including position nt1653, nt1726, nt1727, nt1730, nt1753, nt1762 and nt1764. (3)Nucleotide mutations [8%(5/60)] resulted in amino acid substitution in control group. Nucleotide mutations changed in position nt1632 and nt1635, located in non-functional region.    Conclusions     HBV X gene mutations and the subsequent amino acid substitutions are found in HBV-GN. The urine protein excretion level increases in patients with X mutation, suggesting that these mutations may play an important role in the pathogenesis of HBV-GN.

Objective    To investigate the effects of low density lipoprotein receptor (LDLr) pathway on podocyte injury in diabetic nephropathy (DN) under inflammatory stress.     Methods    Male db/db mice and db/m mice were randomly divided into four groups (8 mice in each group): db/m group (control), casein injected db/m group (db/m＋casein), db/db group (db/db), and casein injected db/db group (db/db＋casein). An inflamed model of DN was established according to our previous study. 24-hour urinary protein was measured every week. The plasma lipid profile was detected by clinical biochemistry assay. Podocyte changes were evaluated by electron microscope and immunofluorescent staining. Lipid accumulation in the kidney was evaluated by oil red O staining and intracellular cholesterol quantitative assay. The protein expression of Wilm's tumor-1 (WT-1), nephrin, α-smooth muscle actin (α-SMA), and molecules correlated with LDLr pathway were examined by immunohistochemical staining or Western blotting. The colocalized protein expression of LDLr with WT-1 was examined by immunofluorescent staining and laser confocal microscopy.    Results    There were no differences in plasma levels of LDL and HDL among four groups. Compared with db/db group, the db/db+casein group showed markedly increased 24-hour urinary protein, more significant podocyte foot process effacement and podocyte damage, increased lipid droplet accumulation in kidneys, increased protein expressions of LDLr, SCAP and SREBP-2 in kidneys (all P＜0.05). Interestingly, increased LDLr protein expression in kidneys of db/db mice was negatively correlated with decreased nephrin protein expression (r＝-0.855, P＜0.01) and positively correlated with increased α-SMA protein expression (r＝0.768, P＜0.01).    Conclusions    The disruption of LDLr pathway induced by inflammation contributes to podocyte injuries in diabetic nephropathy.

Objective    To observe the members’ dynamic change of renin-angiotensin system(RAS) and angiotensin converting enzyme 2(ACE2)-Ang(1-7)-Mas axis during the continuous veno-venous hemodiafiltration (CVVHDF) treating the dogs with multiple organ dysfunction syndrome (MODS), and to investigate the efficacy mechanism on cardiac function.    Methods   Dogs were subjected to hemorrhagic shock plus resuscitation and endotoxemia to establish MODS model, then they were randomly divided into 2 groups: CVVHDF group (n=8) and MODS group (n=6). After endotoxin injection completion, the CVVHDF group received CVVHDF for 12 h, MODS group didn't. Radioimmunoassay, euzymelinked mmunosorbent assay (ELISA) were used to detect the content of renin, AngⅠ, AngⅡ, Ang (1-7). Real-time PCR was used to detect the expression of renin mRNA and ACE2 mRNA. Western blotting was used to detect the protein content of renin, AngⅡ, ACE2 and Ang (1-7).     Results     (1) Organ function: Compared with the MODS group, the vital signs, heart, lung and renal function were significantly ameliorated in the CVVHDF group, the difference had statistical significance (P＜0.05). (2) RAS changes: To detect index from the level of organs, genetic and molecular in arterial tissue, the results displayed that the content of renin, AngⅠ, AngⅡ, the expression of renin mRNA, the protein content of renin, AngⅡ in CVVHDF group were lower than that in MODS group, the difference had statistical significance (P＜0.01). (3)ACE2-Ang(1-7)-Mas axis's changes: Using the same methods above to detect corresponding indicators, the results displayed that the content of ACE2, Ang(1-7), the expression of ACE2 mRNA and the protein content of ACE2, Ang(1-7) in CVVHDF group were significantly improved than that in MODS group, the difference had statistical significance (P＜0.01).    Conclusions    In the process of CVVHDF treating MODS, the ACE2-Ang(1-7)-Mas axis plays the role which antagonizes the RAS system, and to protect the cardiac function. This research may be important for MODS’ clinical rescue.

Objective    To observe the relationship between protein phosphatase-2Ac (PP2Ac) and renal interstitial fibrosis, and to investigate the effects and the underlying mechanism of norcantharidin (NCTD) on renal fibrosis in unilateral ureteral obstruction (UUO) rats.    Methods    Sixteen male Sprague-Dawley rats were randomly divided into four groups: Sham operation group, UUO day 3 group, UUO day 7 group and UUO day 14 group. The rats were sacrificed at day 1, 3, 7, 14 respectively and kidneys were harvested. Immunohistochemical staining were used to detect the expression of fibronectin (FN), type I collagen (Col-I) and PP2Ac. Another twenty male Sprague-Dawley rats were also randomly divided into four groups: Sham operation group, UUO group, low dose NCTD treatment group (0.05 mg·kg-1·d-1) and high dose NCTD treatment group (0.1 mg·kg-1·d-1). The rats in NCTD treatment groups were injected with norcantharidin into abdominal cavity 1 day before operation, while the Sham and UUO group were injected with equal normal saline. Rats were sacrificed at day 14 after surgery and the kidneys were harvested. Immunohistochemical staining, Western blotting and real-time PCR were used to detect the expression of FN, Col-I, α-SMA, E-cadherin and PP2Ac.     Results    (1)Compared with sham group, the expression of PP2Ac, FN and Col-I increased with the development of ureteral obstruction (all P＜0.05). The expression of PP2Ac was positively correlated with the expression of FN and Col-I (r＝0.894 and 0.887, all P＜0.05). (2)Compared with sham group, the expression of FN, Col-I, α-SMA increased and the E-cadherin decreased in UUO group, the expression of PP2Ac also increased (all P＜0.05). After the treatment of NCTD, the above changes were all alleviated in a dose dependent manner, and the expression of PP2Ac was down-regulated (all P＜0.05).    Conclusion    NCTD can ameliorate tubulointerstitial fibrosis and its anti-fibrosis effect may be related to its inhibition to PP2Ac.

Objective To explore the role of liver X receptor (LXR) agonist T0901317 on thrombomodulin (TM) expression in human glomerular endothelial cells and the possible mechanisms. Methods    Different concentrations of T0901317 were used to stimulate human glomerular endothelial cells for different time, then LXRα, LXRβ expression were detected by using Western blotting analysis; the roles of T0901317 on TM mRNA and TM protein expression were observed by using real-time PCR, Western blotting and immunofluorescence assay. LXRα, LXRβ gene interference segment Si-hLXRα, Si-hLXRβ were transfected into human glomerular endothelial cells with the concentration of 100 nmol/L respectively, then the roles of Si-hLXRα, Si-hLXRβ on the TM protein and TM mRNA expression were assayed by Western blotting and real time PCR.    Results    Human glomerular endothelial cells expressed LXRα and LXRβ. Compared to the normal cells and DMSO group, T0901317 could significantly promote TM expression in human glomerular endothelial cells (P＜0.05) and showed a time- and dose-dependent manner. TM expression in Si-hLXRα transfected group was significantly lower than that in the control group (P＜0.05), while TM expression in Si-hLXRβ transfected group had no significant difference compared to the control group.    Conclusions    Human glomerular endothelial cells express LXRα and LXRβ. LXR agonist T0901317 promotes TM expression in human glomerular endothelial cells, which may be mainly through activating LXRa.

Objective    To investigate the role of JAK2-STAT3 pathway in the expression of complement factor B (CFB) in autosomal dominant polycystic kidney disease (ADPKD).    Methods    Renal tissue samples of patients with ADPKD after nephrectomy were collected. Normal renal tissue samples as control were taken from patients after radical nephrectomy. Renal tissue samples of Han: SPRD Cy/+ rats (ADPKD model) and wild-type Han: SPRD +/+ rats were also collected at 4, 8, 16 week. Han:SPRD Cy/+ rat renal tubular epithelial cells (16 w) were primarily cultured in vitro, then stimulated with the JAK2 inhibitor (WP1066) and STAT3 inhibitor (pyrimethamine) for 24 h respectively. Western blotting was used to detect the expression of p-JAK2, JAK2, p-STAT3, STAT3, CFB protein.    Results    Compared with control group, the protein expressions of p-JAK2, p-STAT3, STAT3, CFB significantly increased in the renal tissue of ADPKD patients (all P＜0.05). The protein expressions of p-JAK2, JAK2, p-STAT3, STAT3 and CFB also significantly increased in the renal tissue of Cy/+ rats compared with wild-type rats (all P＜0.01). When the Cy/+ renal tubular epithelial cells were treated with WP1066, the expressions of p-JAK2, p-STAT3, CFB were suppressed (P＜0.05) and the degree of inhibition was correlated with the WP1066 dose. Pyrimethamine inhibited the protein expressions of p-STAT3 and CFB in the tubular epithelial cells of Cy/+ rats (all P＜0.05) and the degree of inhibition was correlated with the pyrimethamine dose.    Conclusions    The JAK2-STAT3 pathway is abnormally activated in ADPKD and increases the protein expression of CFB. CFB protein level is correlated with the progress of ADPKD, suggesting that it may take part in the growth and development of ADPKD vesicles.