Objective    To explore the clinical significance of the serum and urine angiopoietin (ANGPTL) 3 and 4 levels in children with primary nephrotic syndrome (PNS).    Methods    Serum and urine samples from 180 children with PNS admitted from September 2012 to August 2013, and from 18 healthy children as control, were analyzed. Serum and urine ANGPTL3 and 4 concentrations were detected by ELISA. Urine protein (Up), urine creatinine (Ucr), serum creatinine (Scr), blood urea nitrogen (BUN), triglyceride (TG) and total cholesterol (TC) levels were analyzed by automatic biochemical analyzer. Data were analyzed by SPSS 19.0.    Results    (1) Serum ANGPTL3 concentration in PNS children was higher than that in healthy children population [1210.95 (671.28-1571.87) μg/L vs 308.20 (230.05-372.26) μg/L, P＜0.01]; Urine ANGPTL4/Cr in PNS children was also higher than that in healthy children [115.57 (26.50-129.81) ng/g vs 11.26 (2.23-15.11) ng/g, P＜0.01]; Serum ANGPTL4 concentration and urine ANGPTL3/Cr in PNS children had no difference with  those in healthy children population. (2) ANGPTL3 serum levels were positively correlated with age at onset (r=0.199, P=0.047), duration of disease (r=0.501, P=0.027), 24 h urine protein excretion (r=0.384, P=0.004), Up/Ucr (r=0.367, P=0.006), TG (r=0.314, P=0.021), and TC (r=0.444, P=0.001), while controlling the serum lipid level, serum ANGPTL3 level was also correlated with 24 h urine protein excretion (r=0.348, P＜0.001) and Up/Ucr (r=0.312, P＜0.001); Urine ANGPTL4/Cr was positively correlated with 24 h urine protein excretion (r=0.318, P=0.019), Up/Ucr (r=0.117, P=0.044). (3) No difference of serum and urine ANGPTL3 and 4 levels were found among steroid-sensitive, steroid-resistant, steroid-dependent PNS groups, as well as between the frequency relapse or non-frequency relapse groups, while in steroid-dependent and frequency relapse NS children, serum ANGPTL3 level could reflect the curative effect on proteinuria. (4) In PNS children with similar degree of proteinuria and lipid levels, the serum ANGPTL3 levels varied in different pathological types.    Conclusions    Serum ANGPTL3 level may be an important indicator of PNS incidence, disease severities, pathological types and curative effect on proteinuria in PNS. Urine ANGPTL4 level is correlated with proteinuria, which may have the clinical reference value in PNS.

Objective    To explore the relationship between serum sclerostin level, and mineral metabolism, bone density, abdominal aortic calcification in maintenance hemodialysis (MHD) patients.    Methods    Serum sclerostin levels from 175 cases of MHD patients were measured by ELISA. Calcaneus bone mineral density (BMD) was measured by quantitative ultrasound (QUS). The abdominal aortic calcification was detected by abdomen lateral plain radiographs. Interrelations among above parameters were examined statistically.    Results    The median sclerostin concentration of 175 patients was 160.50(100.67, 256.39) pmol/L. Serum sclerostin levels were correlated positively with age, BMI, serum calcium and serum 25(OH)-vitamin D, while negatively with spKt/v and serum iPTH. In multiple regression analysis, serum sclerostin levels were associated significantly and independently with age, sex, BMI and serum iPTH. Compared to patients with normal BMD (T score≥-1s), the patients with low BMD (T score<-1s) had lower serum sclerostin level [142.97(99.52, 226.02) vs 201.13(107.40, 327.84) pmol/L, P=0.035]. Serum sclerostin levels were correlated significantly and positively with calcaneus BMD. Multivariate logistic regression analysis showed that serum sclerostin level was an independent protective factor for low BMD in MHD patients[OR=0.241, 95%CI (0.078, 0.749), P=0.014].    Conclusions    Serum sclerostin levels are associated with mineral disorder and bone density. Sclerostin may become a promising marker of bone turnover in MHD patients.

Objective    To explore the effect of CYP3A5 gene polymorphisms on the whole blood trough concentration (C₀) of tacrolimus (TAC) in patients with idiopathic membranous nephropathy to identify an economical and optimal initial dosage delivering the best curative effect with  minimum drug adverse reaction.    Methods    Sixty patients with idiopathic membranous nephropathy were enrolled in this study. The CYP3A5 genotype was tested by fluorescence in situ hybridization (FISH). According to CYP3A5 genotype, the patients were divided into three groups (AA, AG, and GG). At the same time, the C₀ of TAC was measured by  enzyme multiplied immunoassay technique (EMIT). C₀ of TAC, daily dosage of TAC and the concentration/dose(C₀/D) ratio of TAC were detected after taking medicine at 8, 12, 16 and 24 weeks respectively, so as to corroborate the relation between CYP3A5 gene polymorphisms and the dosage of TAC.    Results    The oral TAC dosage had great variation among individuals. The occurrence of the CYP3A5 genetic polymorphisms (A6986G) designated as G was 53.33%. D and C₀ were significantly different at 8, 12, 16 and 24 weeks respectively (all P＜0.05). To reach the same C₀, the patients with AA needed 2-3-fold dosage of TAC than GG; and those with AG needed 1-2-fold dosage of TAC than GG. After 24-week treatment, the effective rate of AA group was markedly lower than AG and GG (16.67% vs 81.25%, 16.67% vs 87.50%，all P＜0.001). Among CR, PR and NR, there were no significantly difference on C₀ or C₀/D of TAC (P＞0.05).    Conclusions    CYP3A5 genotypes are correlated with blood concentration of TAC. CYP3A5 genotyping may be a new approach to predict the optimal initial dosage of tacrolimus in idiopathic membranous nephropathy.

Objective    To describe the clinical characteristics of one child with primary hyperoxaluria types Ⅲ, and to analyze the potential mutant genes in his family.    Methods    AGXT, GRHPR and HOGA1 genes were analyzed by direct sequencing analysis in this family. One hundred unrelated healthy subjects were also analyzed as controls.    Results    The child had early onset of symptoms (0.8 year). His principal clinical manifestation included nephrolithiasis and obstructive nephropathy, however his nephrocalcinosis was mild. And he presented high urine oxalate, high urine calcium, and lower citrate levels. Two novel heterozygous mutations in HOGA1 were identified (compound heterozygous), one mutation was a 2-bp substitution at the last position in exon 6 and the first position of intron 6 respectively (c.834_834+1GG＞TT); another was a guanine to adenine substitution of the last nucleotide of exon 6 (c.834G＞A). Both of these variants found in this study probably acted as splicing mutations. Direct sequencing analysis failed to find these mutations in 100 unrelated healthy subjects. In addition, a SNP (c.715G＞A, p.V239I) was found in this family. There were no mutations detected in AGXT and GRHPR.    Conclusions    Two novel mutations are identified probably in association with PHⅢ. This is the first description and investigation on mutant gene analysis of PHⅢ in Asia.

Objective    To explore the clinical characteristics and related risk factors in patients with severe uremic secondary hyperparathyroidism (SHPT) complicated with Sagliker syndrome (SS).    Methods    A total of 229 patients with severe uremic SHPT admitted in our hospital from February 2011 to April 2015 were enrolled, among which 33 cases were taken as positive group (SS group), and 196 cases as control group. The differences between two groups in demographic data (such as gender and age), complications, and biochemical indexes were compared, with potential risk factors of SS being analyzed.    Results    There were significant differences between median duration of dialysis in positive group (11 years) and that in control group (8 years, P＜0.001). Compared to control group, the patients in SS group had lower levels of serum creatinine (Scr), blood urea nitrogen (BUN), albumin (Alb), phosphorus (P), and higher serum levels of parathyroid hormone (iPTH), ferritin, hypersensitive C-reactive protein and alkaline phosphatase (ALP), as well as higher calcification in heart valves and abdominal aortic (all P＜0.05). The unadjusted logistic regression models showed that longer duration of dialysis (＞10 years, OR=6.182, P=0.002), higher serum levels of ALP (＞347 U/L, OR=5.786, P=0.002) and iPTH (＞1764 ng/L, OR=4.960, P=0.001), and calcification in heart valves and abdominal aortic (OR=8.635, P＜0.001; OR=5.039, P=0.001) were associated with increased risks of SS, and higher serum Alb was a protect factor for SS (OR=0.904, P=0.014). The multivariate regression analysis showed that longer duration of dialysis (＞10 years, OR=5.121, P=0.036), higher serum level of iPTH (＞1764 ng/L, OR=4.130, P=0.017), calcification in heart valves (OR=11.714, P＜0.001) were independent risk factors of SS.   Conclusions    Severe uremic SHPT patients with longer duration of dialysis and higher serum level of iPTH are more likely to develop SS.

Objective    To compare the performance of newly developed Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation and Modification of Diet in Renal Disease (MDRD) equation in patients with peripheral arterial diseases (PAD).    Methods    A total of 841 patients with PAD were enrolled in this retrospective cohort study. Estimated glomerular filtration rate (eGFR), calculated by MDRD and CKD-EPI equation respectively, was analyzed by Spearman correlation analysis, Bland-Altman method and Kappa test for the evaluation of correlation and consistency. Net re-classification improvement (NRI) was adopted to compare the death risk assessment between these two equations.    Results    Although the eGFR was 4.33 ml•min^-1•(1.73 m²)^-1 higher with MDRD equation than with CKD-EPI equation, there were still significant correlation and fine consistency between eGFR_MDRD and eGFR_CKD-EPI (Kappa：0.749, r=0.991, P＜0.05). The CKD-EPI equation re-classified 9 (1.1%) patients upward to higher eGFR category and 143 (17.0%) patients  downward to lower eGFR category. Besides, the performance of risk assessment for all-cause death was better with CKD-EPI equation than with MDRD equation (NRI=0.059, P＜0.05), which was not the case for cardiovascular death (NRI=0.022, P＞0.05).    Conclusions    There is no solid evidence suggesting that CKD-EPI equation performs better than MDRD equation.

Objective  To investigate the effect of albumin on expression of NLRP3 inflammasome and its downstream cytokines IL-1β and IL-18 in tubular epithelial cells.    Methods  Thirty mesangioproliferative glomerulonephritis (MsPGN) patients with different levels of proteinuria were selected, and their renal biopsy samples were stained by PAS and Masson to observe tubular epithelial cells injury and inflammatory cells infiltration. NLRP3, caspase-1, IL-1β and IL-18, as well as different inflammatory cells, were detected by immunohistostaining.  In vitro, Western blotting and real-time PCR were employed to detect NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA in HK-2 cells stimulated by bovine serum albumin (BSA) (20 g/L).    Results    In MsPGN patients with high levels of proteinuria, there were obvious renal tubular epithelial cell injury and inflammatory cells infiltration (all P＜0.05), and the expressions of NLRP3, caspase-1, IL-1β and IL-18 were up-regulated compared to patients with low levels of proteinuria (all P＜0.05).  Furthermore, IL-1β and IL-18 expressions were positively correlated with the degree of proteinuria (r=0.836, P＜0.05; r=0.901, P＜0.05). NLRP3, caspase-1, IL-1β and IL-18 protein and mRNA were significantly increased in HK-2 cells stimulated by BSA compared to the control group (all P＜0.05).    Conclusions    Albumin is able to induce NLRP3 inflammasome activation in tubular epithelial cells, which may be the mechanism of tubulointerstitial injury and inflammation caused by proteinuria.

Objective    To evaluate the effect of oxidized LDL (Ox-LDL) on scavenger receptor A (SR-A) expression in mouse podocytes and to explore the possible mechanism.    Methods    The conditionally immortalized mouse podocyte cell line was cultured in vitro and exposed to Ox-LDL of different concentrations for 24 h, or 20 mg/L Ox-LDL for different hours. Cell cholesterol accumulation was examined. Real-time quantity PCR and Western blotting were used to analyze the expression level of PTEN, SR-A and nephrin. Podocytes were incubated with DiI labeled Ox-LDL for 4 h and immunofluorescence was used to analyze lipid uptaking. To further confirm the relationship between PTEN and SR-A, PTEN inhibitor bpv (hOpic) [dipotassium bisperoxo (5-hydroxypyridine-2-carboxyl) oxovanadate (V) ], PTEN siRNA and PTEN adenovirus (adPTEN) were used to dual-directional regulate PTEN expression, so as to observe the change of SR-A, nephrin, cell cholesterol accumulation and lipid uptake.    Results    SR-A was expressed on mouse podocyte and mediated podocyte lipid uptake. Compared with control group, Ox-LDL increased cell cholesterol accumulation, and up-regulated SR-A expression along with inhibited expression of PTEN and nephrin (P＜0.05), which were correlate with dose and exposure time of Ox-LDL. Expression of PTEN significantly inhibits the expression level of SR-A and lipid uptake induced by Ox-LDL (P＜0.05), thereby decreasing cell cholesterol accumulation, but up-regulating nephrin level (P＜0.05). However, down-regulation of PTEN could cause opposite effect.    Conclusion    Ox-LDL up-regulates SR-A through decreasing the expression of PTEN, and contributes to podocyte injury.

Objective    Long noncoding RNA (lncRNA) has been identified to regulate DNA methylation, histone acetylation, and gene post-transcriptional regulation in kinds of diseases, including tumorigenesis, obesity and so on. Therefore, lncRNAs might be the potential targets of mesangial cells proliferation.    Methods    Mesangial cells were exposed to suitable concentration of TGF-β through cell proliferation assay; then the lncRNAs expression levels were detected by microarray in experimental group and control group separately; finally the differentially expressed lncRNAs were identified by      RT-PCR; meanwhile, and the expression levels of target genes were also detected by RT-PCR.    Results   Cell viability assay confirmed that 10 ng/ml TGF-β could promote mesangial cell proliferation significantly. Totally, over 30 000 lncRNAs were detected in TGF-β treated MCs and control group cells separately. Compared to the control group, 5550 lncRNAs differentially expressed in TGF-β treated MCs, including 119 up-regulated and 147 down-regulated over 2 fold. RT-PCR results appeared that uc.60, MRAK079149, MRAK029456, XR_005507, XR_007641, uc.14, and uc.412 were significantly up-regulated in TGF-β treated MCs, and BC088254, DQ402472, BC098733, BC158832, BC098746 were stably down-regulated. Compared to the control group, the mRNA expression levels of AATF and NEK were increased in the TGF-β treated mesangial cells (P＜0.05). AATF and NEK were downstream target genes of uc.412 and MRAK079149 respectively.    Conclusion    The differential expression of long noncoding RNAs presents in the experimental mesangial cells proliferation induced by TGF-β.