Objective   To conduct a prospective, randomly controlled trial, evaluating the combination of tacrolimus, corticosteroids and entecavir for the treatment of adult patients with biopsy-proven hepatitis B virus - associated membrane nephropathy (HBV - MN).  Methods    A total of 38 patients with biopsy - confirmed HBV - MN were randomized to the tacrolimus group (n=19) and the control group (n=19). Patients in tacrolimus group received combination therapy of tacrolimus (0.05 mg•kg^-1•d^-1), corticosteroids (prednisone acetate, 0.5 mg•kg^-1•d^-1) and entecavir (0.5 mg/d), whereas patients in control group received entecavir mono-therapy (0.5 mg/d). The primary end point was the percentage of patients reaching complete remission (CR) or partial remission (PR).  Results    The probability of remission in the treatment group was 88.89% and 94.44% after 6 and 12 months, but only 38.89% and 58.82% in the control group, respectively. The decrease in proteinuria was significantly greater in the treatment group. Entecavir was used for the treatment of hepatitis in all patients, which caused the disappearance of serum hepatitis B viral DNA(HBV-DNA) and the normalization of ALT and AST levels in 3 months. Notably, two patients in the control group and one patient in the treatment group reached the secondary end point. One patient in the tacrolimus-treated group showed a relapse during the taper period.  Conclusion   This treatment protocol not only can control the replication of HBV-DNA but also can reduce proteinuria and preserve renal function, it is one of useful therapeutic options for patients with HBV-MN.

Objective   To analyze the clinical features of renal and urinary lesions in IgG4 related disease (IgG4 - RD).  Methods   Clinical data of 24 cases of IgG4 - RD patients were retrospectively analyzed, including serum creatinine, hemoglobin, inflammatory factors, and serum IgG4 level before and after treatment.  Results   Twenty four cases of IgG4 - RD with renal and urinary lesions were diagnosed in Peking Union Medical College Hospital from Aug 2010 to May 2014, including 21 males and 3 females. The average age of the patients was (63.79±12.13) years old, and the average number of organs involvement was (4.38±1.55). Elevated inflammatory factors and serum IgG4 level were detected in all patients. Elevated urine protein was detected in 19 cases and 6 cases were diagnosed as nephritic syndrome among 20 patients. Increased Scr were detected in 21 cases with an average value of (410.48±352.17) μmol/L and 3 cases had been treated with hemodialysis. Anemia was found in 14 cases and 8 cases were detected with single or bilateral kidney atrophy. Among the 21 cases with elevated Scr, 11 cases were caused by postrenal obstruction and 3 cases were pathologically proved as tubulointerstitial nephritis. Received steroid treatment, Scr level declined dramatically one week later (P＜0.05) and further declined one month later (P＜0.01); erythrocyte sedimentation rate declined significantly one month later (P＜0.01)；serum IgG4 level did not decline significantly after one month (P=0.146), but declined significantly after two months (P＜0.01).  Conclusions   Renal insufficiency is not rare in IgG4 - RD with renal and urinary lesions, and it is sensitive to steroid therapy; the decline in Scr can be seen earlier than the change in inflammatory factors and serum IgG4 level.

Objective   To observe the effects of parathyroidectomy(PTX) on bone metabolism and bone mineral density(BMD) in maintenance hemodialysis patients with secondary hyperparathyroidism (SHPT).  Methods   A total of 26 dialysis patients with SHPT were treated with PTX. Serum calcium, phosphorus, alkaline phosphatase(ALP) levels were determined by standard methods. The levels of serum intact parathyroid (iPTH), osteocalcin (OC), procollagen type I amino- terminal propeptide (PINP), β - crosslaps (β - CTX) were measured by chemiluminescence. BMD was measured by dual energy X ray absorptiometry. iPTH, OC, PINP, β-CTX, serum calcium, phosphorus, ALP were measured before parathyroidectomy and 1, 3, 6, 12, 18, 24 months after operation. Lumbar spine (LS) and femoral neck (FN) BMD were measured before and 24 months after PTX.  Results   Compared with that before operation, serum OC[(104.49±25.42) μg/L vs (695.46±355.62) μg/L, P＜ 0.01] and PINP levels [(248.36 ± 159.38) μg/L vs (809.28 ± 283.50) μg/L, P＜0.01] progressively decreased 3 months after PTX, and serum β-CTX levels [(1.60±0.64) μg/L vs (3.37±1.34) μg/L, P＜ 0.01] decreased 1 month after PTX. Compared with that before operation, BMD levels increased 24 months after PTX in LS[(0.88±0.23) g/cm² vs (0.78±0.23) g/cm², P＜0.01] and FN[(0.96±0.19) g/cm² vs (0.84±0.24) g/cm², P＜0.01], and Z-scores were also increased in both LS[(-1.24±0.55) vs (-1.66± 0.24), P＜0.01] and FN[(-1.51±0.72) vs (-1.93±0.40), P＜0.01]. Correlation analysis showed that baseline iPTH was positively correlated with △Z-score in FN (r=0.584, P=0.002) and LS (r=0.400, P= 0.043), and so did the OC with △Z - score in FN (r=0.651, P＜0.001) and LS (r=0.509, P=0.008).  Conclusion   The levels of OC, PINP and β-CTX are reduced and BMD is improved in hemodialysis patients with SHPT after PTX.

Objective   To investigate the relationship between the prevalence of isolated systolic hypertension (ISH) and the stages of chronic kidney disease (CKD) in chronic kidney disease outpatient clinic.  Methods   CKD patients of stages 1, 2, 3, 4 and 5 were recruited (n=626). Based on office systolic pressure (SBP) and diastolic pressure (DBP), they were classified into four subtypes: normotension (＜140/90 mmHg), isolated diastolic hypertension (IDH，SBP＜140 mmHg and DBP ≥ 90 mmHg), ISH (SBP≥140 mmHg and DBP＜90 mmHg) and systolic- diastolic hypertension (SDH, SBP≥140 mmHg and DBP≥90 mmHg).  Results   The control rate of blood pressure was 86.4%, 75.6%, 65.3%, 51.0% and 37.0% at CKD stage 1, 2, 3, 4 and 5, respectively, which decreased with the advancement of CKD. There was a stepwise increase in the prevalence of ISH (0, 9.2%, 23.9%, 28.6% and 37.0% at CKD stage 1, 2, 3, 4 and 5, respectively) and SDH (4.5%, 8.4%, 8.0%, 17.3%, 21.9% at CKD stage 1, 2, 3, 4 and 5, respectively). Logistic regression analysis showed that age, diabetes and CKD stages were independent predictors of ISH. Compared with CKD stage 1-2, CKD stage 3, 4 and 5 showed 2.388, 2.697 and 5.980 folds risk in developing ISH.  Conclusion  The prevalence of ISH increases correspondingly with the advancement of stages of CKD, which may partially contribute to the increased cardiovascular mortality during the progress of CKD.

Objective    To evaluate the association between serum 25-hydroxyvitamin D₃ [25(OH)D₃] and arterial stiffness in patients with chronic kidney disease (CKD).  Methods    Three hundred patients with CKD were included, and were divided into two groups based on serum 25(OH)D₃ levels: vitamin D deficient [25(OH)D₃＜20 μg/L] and vitamin D non-deficient [25(OH)D₃≥20 μg/L]. Brachial ankle pulse wave velocity (baPWV), which reflected arterial stiffness, was calculated using the single-point method. Clinical data were collected in details. Correlation between serum 25(OH)D₃ level and baPWV was assessed by the single factor correlation test and multiple linear regression analysis.  Results    The prevalence of vitamin D deficiency was 62.7%(188/300). The concentration of 25(OH)D₃ was (17.62±8.54) μg/L in total patients, but was (12.38±4.55) μg/L and (26.44±6.05) μg/L in the subgroups of vitamin D deficient and non-deficient, respectively(P＜0.01). There was a higher value of baPWV in the group of vitamin D deficient than that of vitamin D non-deficient (1 827.34±429.11 vs 1 555.31±353.14, P＜0.01). Serum 25(OH)D₃ level and baPWV was negatively correlated in total patients(r=-0.38, P＜0.01) and each stage of CKD(stage 2-5)[r=-0.30, P＜0.05; r=-0.26, P＜0.05; r=-0.46, P＜0.01; r=-0.57, P＜0.01]. Multiple linear regression analysis showed that vitamin D level was independently associated with baPWV(Model 1: β =-0.18, P＜0.01; Model 2: β =-0.17, P=0.01). Both models accounted for 50% (R²=0.50) of total variance of baPWV.  Conclusions    Vitamin D deficiency is common in CKD, and a low 25(OH)D₃ level is significantly associated with increased arterial stiffness in these patients. Clinical intervention studies are needed to clarify whether treatment with vitamin D decreases the risk of cardiovascular disease in patients with CKD.

Objective    To systematic evaluate the efficacy of paricalcitol on estimated glomerular filtration rate (eGFR) and proteinuria in non-dialysis chronic kidney disease (CKD) patients. Methods    According to the collaborative search strategy, PubMed, the clinical control test database of Cochrane Library, Embase, Chinese Wanfang database, CNKI, VIP database (form the date of database establishment to March 2014) were searched. Published and unpublished literature, abstracts in academic meetings (ASN, WCN, CSN) were also searched by hand. The randomized controlled trials (RCTs) about the efficacy paricalcitol on eGFR and proteinuria in non - dialysis CKD patients were selected. Review Manager Software 5.2 was used for statistical analysis. Results    Seven RCTs with a total of 834 patients were included (508 in experimental group, 326 in placebo group). No statistical difference of the efficacy on eGFR[SMD=-0.10, 95% CI: (-0.28 - 0.07), P=0.26] between lower dose paricalcitol (＜2 μg/d) group and placebo group, while higher dose (2 μg/d) group reduced eGFR significantly [SMD=-0.45, 95% CI: (-0.63 - -0.27), P＜0.01]. Compared with placebo, paricalcitol reduced proteinuria significantly [OR(95%CI): 2.09(1.52-2.58), P＜0.01], and there was no difference between different dose groups [OR(95%CI): 1.09(0.62-1.91), P=0.77]. Lower dose group [OR(95%CI): 0.93(0.57 - 1.52), P=0.76] and higher dose group [OR(95% CI): 2.08(0.70 - 6.18), P=0.19] did not significantly increase the risk of adverse events.  Conclusions    Lower dose paricalcitol (＜2 μg/d) has no effect on eGFR in non-dialysis CKD patients meanwhile reduces proteinuria. The higher dose (2 μg/d) may reduce eGFR without farther reduction in proteinuria.

Objective    To explore the effect and mechanism of magnesium on calcification induced by hyperphosphate.  Methods    Vascular smooth muscle cells (VSMCs) were primarily cultured in vitro and induced calcification by β - glycerophosphate (β - GP). VSMCs were randomly divided into control group, high phosphorus group (10 mmol/L β-GP), magnesium intervention group(10 mmol/L β - GP + 3 mmol/L MgSO₄) and 2 - aminoethoxy - diphenylborate (2 - APB, an inhibitor of magnesium transporter) intervention group(10 mmol/L β-GP+3 mmol/L MgSO₄ +10^-4 mol /L 2-APB). Calcium deposition and alkaline phosphatase (ALP) activity were measured by alizarin red staining, quantification of calcium and enzyme linked immunosorbent assay. RT-PCR and Western blotting were used to observe the expression of core binding factor α-1 (Cbfα-1) mRNA and protein, respectively. In vivo, male Sprague-Dawley rats (n=24) were randomly divided into control group (methylcellulose+high phosphorous diet), vascular calcification group (adenine suspension + high phosphorous diet), high magnesium intervention group(adenine suspension+high phosphorous and magnesium diet). The aortic pulse wave velocity (PWV) was measured, and vascular calcification was determined by von Kossa stain and quantification of calcium. Cbfα-1 in aortic was measured by immunohistochemistry.  Results    In vitro, compared with high phosphorus group, calcification, ALP activity (P＜0.05) and Cbfα - 1 expression in VSMCs were significantly decreased in magnesium intervention group after incubation for 14 days, but the addition of 2 - APB might inhibit the protective effect of magnesium on VSMCs. Dynamic observation of Cbfα-1 showed that magnesium significantly inhibited the expression of Cbfα-1 (P＜0.05) on the third day and the inhibitory role was obviously increased in a time - dependent manner. Consistent with the findings in vitro, the aortic PWV, calcification were all significantly reduced (P＜0.05) in high magnesium intervention group with high serum magnesium level, when compared with vascular calcification group. Immunohistochemistry showed that hypermagnesemia down- regulated obviously the expression of Cbfα-1 induced by hyperphosphatemia(P＜0.05).  Conclusion    Magnesium protects against vascular calcification by inhibiting osteogenic differentiation of VSMCs.

Objective    To observe the effect of fasudil on cytoskeleton reconstruction in mouse podocytes induced by angiotensinⅡ(AngⅡ), as well as to study the protective mechanism of fasudil in the pathological changes of podocytes. Methods    Conditionally immortalized mouse podocytes were treated with AngⅡ(10^-7 mol/L). The podocytes were pre-incubated for 30 min or 60 min with various concentrations of fasudil (10^-8, 10^-7, 10^-5 mol/L), then Ang Ⅱ (10^-7 mol/L) were added and further - incubated for 24 hours. The cytoskeleton distribution of podocyte as indicated by F - actin and synaptopodin was observed by fluorescence microscopy and Western blotting. At the same time, the activity of Rho signal pathway that mediates actin filament polymerization was analyzed by measuring the extent of Rho-associated coiled-coil-containing protein kinase 1 (ROCK-1) and myosin phosphatase-1 (MYPT1). Results    Compared to the control group, AngⅡ disrupted the podocyte actin cytoskeleton and significantly decreased the expression of synaptopodin (P＜0.05), while fasudil stabilized the actin filaments, and improved the synaptopodin expression (P＜0.05). The expression enhancement of ROCK-1 and MYPT1 by Ang Ⅱ were reduced significantly by fasudil (P＜0.05). Conclusion    The cytoskeleton reconstruction of podocytes can be induced by AngⅡ and inhibited by fasudil, which suggests that the protective effect of fasudil may be partially contributed to the Rho/ROCK signaling pathway.

Objective    To investigate the underlying mechanism of ursolic acid in attenuating diabetic mesangial cells injury induced by high glucose (HG).  Methods    Rat glomerular mesangial cells were cultured in normal glucose, HG, HG with LY294002 and HG with ursolic acid. The cell proliferation and hypertrophy were detected by MTT and the ratio of total protein content to cell number. miRNA - 21 was detected by quantitative real - time PCR. The PI3K - Akt - mTOR pathway, autophagy associated protein and collagen I were detected by Western blotting and quantitative real- time PCR. The autophagosomes were observed by electron microscope.  Results    Compared with normal control group, the cells exposed to HG showed up-regulating miRNA-21 expression(P＜0.01), down-regulating PTEN protein and mRNA expression(P＜0.01), up-regulating p85PI3K, phospho(p)-Akt, p-mTOR, p62/SQSTMI, collagen I expressions and down-regulating LC3II expression(P＜0.01). Ursolic acid and LY294002 inhibited HG-induced mesangial cell hypertrophy and proliferation(P＜0.01), down -regulated the expressions of p85PI3K, p-Akt, p-mTOR, p62/SQSTMI and collagen I and up-regulated the expression of LC3II(P＜0.01). But LY294002 had no effect on the expression of miRNA-21 and PTEN. Ursolic acid down-regulated miRNA-21 expression(P＜0.01), up-regulated PTEN protein and mRNA expression(P＜0.01).  Conclusion    Ursolic acid may inhibit high glucose-induced mesangial cell miRNA-21 overexpression, up-regulate PTEN, inhibit the activation of PI3K-Akt-mTOR signaling pathway and the enhanced autophagy to reduce the accumulation of extracellular matrix and ameliorate cell hypertrophy and proliferation.