Chinese Journal of Nephrology

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Analysis of CLCNKB gene mutation in a family with classic Bartter syndrome

LIU Mao-jing;YU Ying;GAO Jie;FAN Li;HU Zhao

2011, 27(6): 395-399.

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Objective To investigate the mutations of CLCNKB gene in a family with classic Bartter syndrome. Methods Genetic DNA was extracted from peripheral blood leucocytes of family members. The coding exons and intron exon junctions of CLCNKB gene were amplyfied by PCR and sequenced directly. Fifty unrelated healthy subjects were selected to exclude the possibility of polymorphism. Results A heterozygous (missense) mutation (482T>G, L161R)was detected in the exon 4 of patients. The heterozygous mutation (L161R) was found in the mother, while no mutation was found in the father of this family. L161R had not been reported and was a novel mutation when referring to literatures and human genomic database home and abroad. Conclusion A new CLCNKB gene mutation (L161R) is identified for the first time.

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Prevalence and risk factors of chronic kidney disease in the adult population receiving healthy examination from Xinjiang

FAN Wen-juan;WANG Shu-xia;MINAWAER Yunusi;SANG Xiao-hong

2011, 27(6): 400-405.

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Objective To investigate the prevalence and risk factors of chronic kidney disease (CKD) in the adult population receiving healthy examination from Xinjiang． Methods A total of 10 025 subjects who were older than 20 years and received healthy examination in Health Center of the First Affiliated Hospital, Xinjiang Medical University, from January 2009 to April 2010 were enrolled. Results The prevalence of proteinuria was 4.86％(95％CI:4.44%-5.28%), hematuria 3.13% (95%CI:2.79%-3.47%), and reduced eGFR 2.66%(95%CI:2.34%-2.98%). Approximately 9.60% (95%CI:9.02%-10.18%) of these　subjects had at least one indicator of kidney damage． Albuminuria was independently associated with male, systolic blood pressure, BMI, glycometabolism abnormity, hypertriglyceridemia. Hematuria was independently associated with female, minority, aystolic blood pressure. Reduced eGFR was independently associated with female, age, obesity, hyperlipidemia, hyperuricemia. Age, gender, glycometabolism abnormity, systolic blood pressure, hyperuricemia and hyperlipidemia were independently correlated with CKD. Conclusions The prevalence of chronic kidney disease is 9.60% in the adult population receiving healthy examination, which is quite high and should be considered. The spectrum and correlated factors of CKD in this population are similar to studies reported．

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Investigation of death events caused by infection in end-stage renal disease patients undergoing hemodialysis

TANG Ying;ZHONG Yi-hong;GONG Shao-min;WANG Yi-mei;LV Wen-lv;DING Xiao-qiang

2011, 27(6): 406-410.

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Objective To explore the clinical characteristics and prevention management of death events caused by infections in end-stage renal disease (ESRD) patients undergoing hemodialysis. Methods Clinical data of ESRD patients undergoing hemodialysis in Nephrology Department of Zhongshan Hospital from 1998 to 2008 were retrospectively studied. Death causes, primary diseases, complications, infections, and survival time were analyzed. Results A total of 252 patients died including 162 males（64.29%） and 90 females（35.71%）. Average death age was (63.48±14.77) years. In death events, emergency dialysis accounted for 59.52%, and primary glomerular disease was the major primary diseases (27.23%), then diabetic nephropathy (16.90%) and hypertensive nephrosclerosis (14.55%). 34.8% death was caused by infections or promoted by infections, secondly by cerebrovascular events (23.6%). The elderly accounted for the majority of infection-associated deaths. 48.15% and 38.71% patients with deaths caused or promoted by infections respectively had shorter dialysis duration (<3 months), whose percentage was much higher than those with non-infection-associated deaths (30.34%). Primary infection was still pulmonary infection (77.59%), then blood infection (10.34%) and catheter-associated infection (5.17%). 58.62% infection-associated deaths had positive microbiologic test results, and gram negative bacillus accounted for 38.24%. 50% of patients with positive test was complicated with fungal infection. Conclusions Infection is a main cause and a critical promotion to death in ESRD hemodialysis patients, besides it is the main cause of death in the elderly (>75 years) and hemodialysis duration within 3 months, which may result in shorter survival. Pulmonary infection and gram negative bacillus combined with fungal infection should be considered in the treatment. Prophylaxis of nosocomial infection and pulmonary infection in hemodialysis patients should be more emphasized.

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Association of serum L-kynurenine with nutritional status and vascular stiffness in maintenance hemodialysis patients

WANG Li*;HONG Da-qing;WANG　Fang;LI Ming;WU Shu-kun;GAO Hui;WANG Jun-ru;WANG Zheng-tong;HE Qiang;LI Gui-sen;LIU Zhang-suo;YANG Xiu-chuan

2011, 27(6): 411-415.

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Objective To study the association of serum L-kynurenine changes with nutritional status and vascular stiffness in maintenance hemodialysis (MHD) patients. Methods Twenty gender- and age-matched healthy volunteers (healthy group) and 40 MHD patients, including 20 cases with α-keto aicd (α-keto acid group) and 20 cases without α-keto aicd (non-α-keto acid group) were enrolled in the study. Serum L-kynurenine was measured by high performance liquid chromatography. C-reactive protein (CRP) and interleukin 6 (IL-6) were detected. Subjective global assessment (SGA) and malnutrition inflammation score (MIS) were applied to evaluate the nutritional status. Pulse wave velocity (PWV) was used to evaluate arterial stiffness for both groups of MHD patients. Results Serum L-kynurenine was significantly higher in MHD patients than that in healthy subjects [(3.20±1.12) μmol/L vs (1.74±0.27) μmol/L, P<0.01], while such difference was not found between α-keto aicd group and non-α-keto-aicd group [(3.20±0.88) μmol/L vs (3.29±1.34) μmol/L, P>0.05]. IL-6 was significantly higher in MHD patients as compared to healthy subjects [(6.45±3.78) ng/L vs (1.38±1.59) ng/L, P<0.01], while such difference was found between α-keto aicd group and non-α-keto aicd group [(3.37±0.82) ng/L vs (9.62±2.48) ng/L, P<0.05]. There was no difference of CRP concentration between two MHD groups. As compared to non-α-keto acid group, higher SGA score (26.00±1.75 vs 22.67±2.61, P=0.001), lower MIS score (5.82±2.27 vs 10.00±2.62, P＝0.002), lower left side PWV [(21.11±8.21) m/s vs (24.57±5.45) m/s, P=0.244] and lower right side PWV [(19.27±3.22) m/s vs (24.19±5.41) m/s, P=0.015] were observed in α-keto aicd group. Pearson analysis showed positive correlation between serum L-kynurenine and IL-6 (r＝0.352, P＝0.011) and negative correlation between L-kynurenine and pre-dialysis Scr (r=-0.412, P=0.019). Conclusions Inflammation is common in MHD patients. Tryptophan degeneration product L-kynurenine may indicate inflammation status. α-keto acid improves nutritional status, anemia and arterial stiffness maybe through the alleviation of inflammation in MHD patients.

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Acute kidney injury is a risk factor for the long-term prognosis of cardiac surgery

LIU Shang;CHE Miao-lin;XIE Bo;XUE Song;ZHU Ming-li;QIAN Jia-qi;NI Zhao-hui;YAN Yu-cheng

2011, 27(6): 416-422.

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Objective To investigate the long-term prognosis and possible risk factors of acute kidney injury(AKI) following cardiac surgeries. Methods Clinical data of 941 patients undergoing open heart surgery from January 2004 to June 2007 were retrospectively analyzed. Survival and renal function of above patients were informed through telephone or clinic follow-up till February 2010. Long-term survival was analyzed using risk-adjusted Cox proportional hazards regression model. Results Of the 941 patients after cardiac surgery, 275 (29.2％) developed AKI and 666（70.8%）did not. Survival was worse in patients with AKI（67.8% vs 85.6%，P<0.01）and was proportional to its severity (AKIN 1, 2 and 3 stages: 70.7%, 62.3% and 58.6%，P<0.01). Although 90.5% of patients had complete renal recovery after AKI at discharge of hospital, they still had an increased risk for death during follow-up compared with patients without AKI (69.6% vs 85.6%，P<0.01). In the Cox proportional hazards regression model, age (HR=2.238), anemia (HR=1.625), prolonged operation time (HR=1.153), AKI severity (HR=1.473) were independent risk factors for long-term prognosis after cardiac surgery. At the end of the follow-up, patients with AKI had statistically higher Scr than non-AKI patients (107.6 μmol/L vs 83.0 μmol/L, P=0.014), and among those AKI patients, 34.0% progressed into CKD 3-5 stages, 34.8% developed double serum creatinine. Conclusions AKI is a risk factor for the long-term prognosis of cardiac surgery. Patients with complete renal recovery after AKI still has an increased risk for death and poor renal function compared with patients without AKI.

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Effects of the different sections of receptor-associated protein on the expression and distribution of TRPC6, synaptopodin and podocalyxin in passive Heymann nephritis

CAO Hai-xia;LI Da-yong;FAN Ya-ping;DA Zhan-yun;WANG Feng;CHEN Xiao-lan;WU Ya-jun;SHI Lan

2011, 27(6): 423-427.

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Objective To investigate the effects of different sections of receptor associated protein (RAP) on the expression and distribution of TRPC6, synaptopodin and podocalyxin in passive Heymann nephritis (PHN). Methods Male Sprague-Dawley rats were injected with three kinds of antisera (anti-RAP full-length serum, anti-RAP N-terminal serum and anti-RAP C-terminal serum) to establish three kinds of PHN models. The control group was injected with normal rabbit serum. The quatitation of 24 h urinary protein, serum albumin and creatinine were taken before injection and one week after PHN model successfully induced. The histopathologic changes of renal tissues were observed by light microscopy. The expression and distribution of TRPC6, synaptopodin and podocalyxin in glomerular podocytes were observed by laser scanning confocal microscopy and analyzed by fluorescence quantitative software after indirect immunofluorescence double staining. Results The quantities of 24 h urinary protein in the three model groups were significantly higher than those of themselves before injection and control groups (P<0.01, respectively). The values of serum albumin and creatinine were not significantly different before and after injection (P>0.05). The expression of TRPC6 in podocytes was higher in the PHN model groups than that of control group. Fluorescence intensity of TRPC6 in RAP full-length group was stronger than that in RAP N-terminal or C-terminal groups. The expressions of synaptopodin and podocalyxin distributed along the glomerular basement membrane as spot, discontinuous short line and defect of some segments, and were lower in three PHN groups than those of control group. Fluorescence intensity of synaptopodin and podocalyxin among three PHN groups had no differences. Conclusions RAP full-length and N-terminal or C-terminal parts can increase the expression of podocyte TRPC6, but decrease the expressions of synaptopodin and podocalyxin, and alter their distribution, which may be associated with the proteinuria, however, their role in the PHN pathogenesis needs further study.

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Effects of low protein diet with α-keto acid supplement on kidney sclerosis and renin-angiotensin system in renal ablation rats

LONG Quan;CHEN Ｊing;HAO Chuan-ming;LIN Shan-tan;GU Yong

2011, 27(6): 428-434.

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Objective To observe the influence of low protein diet with α-keto acid on kidney sclerosis and renin-angiotensin system in renal ablation rats. Methods Chronic renal failure rat model was established by renal ablation in 30 male SD rats, then the animals were randomly assigned to the following diet groups: normal protein group (NPD: 18% casein protein), low protein group (LPD: 6% casein protein) and supplemented low protein group (LK: 5% casein protein＋1% α-keto acids). Ten male SD sham-operated rats received 18% casein protein as control. All the rats were killed at the end of the 12th week. Pathologic changes were assessed by PAS staining. AngⅡ in homogenate and plasma were measured by radioimmunoassay and ELISA respectively. Immunohistochemistry and Western blotting were used to detect the protein expression of TGF-β1, renin and AT1R. Real-time PCR was used to detect the gene expression of renin and AT1a, the main subtype of AT1 receptor. Results Body weight, total protein and serum albumin had not significant difference among the four groups (all P>0.05). Serum creatinine and proteinuria of nephrectomized rats were significantly higher compared to the control group (all P<0.05). Proteinuria of the LK group was lower than that of NPD and LPD groups (all P<0.05). Pathological results indicated fibrosis indices were significantly improved after LPD and LK intervention. Expressions of renin, AngⅡ and AT1R in LK group were significantly lower than those in NPD group (all P<0.05). Conclusions Low protein diet with α-keto acids supplement therapy exhibits renal protective effects of reducing urine protein excretion and improving renal fibrosis, which might be related to the attenuation of local renin-angiotensin system in activity nephrectomized rats.

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Change of renin-angiotensin system in cultured mesangial cells by serum from nephrectomized rats feeding with low protein diet and α-keto acid supplement

YIN Ying;LONG Quan;YOU Li;LI Hai-ming;ZHANG Li-yin;GU Yong;HAO Chuan-ming;LIN Shan-tan;CHEN Jing

2011, 27(6): 435-441.

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Objective To observe the changes of renin-angiotensin system（RAS） in cultured mesangial cells by serum from 3/4 nephrectomized rats feeding with low protein diet and α-keto acid. Methods Thirty male SD rats received 3/4 nephrectomy (Nx) were placed on 18% normal protein diet (NPD, n=10), 6% low protein diet (LPD, n=10) or 5% low protein plus 1% α-keto acid diet (LK, n=10) for 12 weeks. Ten male SD sham-operated rats fed with 18% normal protein diet were used as control (sham group). In addition, mesangial cells were cultured in sera (10%) collected from above animals treated with or without losartan (0.02 mmol/L) for 48 hours. ELISA was applied to detect the level of AngⅡ, TGF-β1 and fibronectin(FN) in cell medium. Western blotting was used to determine the protein level of AT1 receptor (AT1R) and real-time PCR was used to detect the mRNA level of AT1R, TGF-β1 and FN. Results (1) Nutritional indices including body weight, total protein and albumin had no significant difference in each group. (2) Serum creatinine and 24 h proteinuria were significantly inceased in nephrectomized groups compared to sham group (P<0.05, respectively). 24 h proteinuria was greatly lower in LK group than that in NPD and LPD groups (P<0.05, respectively). (3) LK greatly decreased the level of AngⅡ[NPD(12.70±0.12) mg/g protein; sham (8.04±0.62) mg/g protein] in supernatant as well as the protein and mRNA expression of AT1R in cultured mesangial cells (P<0.05). (4) NPD serum directly induced higher secretion [FN: sham (20.58±0.46) g/g protein, NPD(39.84±0.06) g/g protein; TGF-β1: sham (10.12±0.56) mg/g protein, NPD(83.85±3.58) mg/g protein] and mRNA expression of FN and TGF-β1 compared with sham group (P<0.05). LPD decreased these increment (P<0.05) and LK showed stronger inhibitory effect (P<0.05). (5) Losartan application sharply reduced FN and TGF-β1 production both in supernatant and in mRNA expression in NPD serum treated cells (P<0.05, respectively). Conclusion Low protein diet with α-keto acids supplement directly inhibits the RAS in mesangial cells which may contribute to its beneficial effect on the kidney.

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Effects of erythropoietin on glomerular endothelial cells function in rats with chronic renal failure

WAN Jian-xin;YANG Xia;CUI Jiong;ZOU Zhen-huan

2011, 27(6): 442-447.

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Objective To investigate the effects of erythropoietin (EPO) on the function of glomerular endothelial cells in rats with chronic renal failure (CRF). Methods The CRF model was established by a two stage 5/6 nephrectomy procedure in rats. Experimental rats were randomly divided into four groups: sham operation group (control group), CRF group, CRF rats treated with 30 U/kg EPO (low-dosage group) and with 50 U/kg EPO (high-dosage group). CRF rats received EPO by hypodermic injection for 6 weeks and then were sacrificed. Serum creatinine (Scr), blood urea nitrogen (BUN), urine protein, haematoglobin (Hb) and blood pressure were measured. The renal morphologic changes were evaluated on periodic acid-schiff (PAS) stained sections. The CD34 and CD31 expressions in glomerulus were detected by immunohistochemistry method. The mRNA of endothelin 1(ET-1), endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor(VEGF) were detected by RT-PCR. Results The expressions of CD34 and CD31 protein in glomerulus, and the expressions of eNOS and VEGF mRNA in renal tissue were higher in EPO treatment group than those in CRF model group (all P<0.05). The expression of ET-1 mRNA in renal tissue was lower in EPO treatment group than that in CRF model group. In addition, the Scr, BUN, urine protein and blood pressure in EPO treatment group were significantly lower than those in CRF model group (all P<0.05). Haematoglobin in EPO treatment group was higher than that in CRF model group (P<0.05). Reanl pathological injury was improved by EPO treatment in dose-dependent manner. Conclusion EPO can ameliorate renal pathological injury and renal function in rats with chronic renal failure, maybe through promoting the renovation of glomerular capillary endothelium and improving the function of glomerular endothelial cells.

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Change of V-ATPase B subunit in transforming growth factor β1-induced epithelial-mesenchymal transition of rat renal proximal tubular cells

CAO Xue-qin;ZHAO Shi-li;QIN Jing;LI Xiao-yan;FAN Jin-jin;MAO Hai-ping;YANG Qiong-qiong;YU Xue-qing

2011, 27(6): 448-453.

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Objective To investigate the change of V-ATPase B subunits on epithelial to mesenchymal transition (EMT) in rat renal tubular epithelial cells (NRK52E) stimulated by transforming growth factor β1 (TGF-β1). Methods NRK52E cells were stimulated by TGF-β1 (10 μg/L) for 0 h (control), 12 h, 24 h, 48 h, 72 h after serum-free culture for 24 h. The mRNA and protein expression of E-cadherin, α-SMA, B2 and B1 subunits of V-ATPase were detected by real-time PCR, Western blotting and immunofluorescence. Results After stimulated by TGF-β1 (10 μg/L) for 48 h, the expression of α-SMA was markedly increased (P<0.05), but the expression of E-cadherin was dramatically decreased(P<0.05). Meanwhile, the expressions of V-ATPase subunit B2 was significantly increased (P<0.05). However, the B1 subunit distributed rarely in NRK 52E cells, and did not increase after TGF-β1 stimulation. Double-label immunofluoerscence staining also showed that the V-ATPase B2 subunit was increased in the cytosol, tending to accumulate to the cell membrane after TGF-β1 stimulation. Conclusions The main isoform of V-ATPase distributed in NRK52E cells is B2 subunit. B2 subunit is increased alone with TGF-β1-induced EMT. It may suggest that V-ATPase B2 subunit may play a potential role in TGF-β1-induced tubular EMT and renal fibrosis.

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临床研究

基础研究

临床研究

基础研究