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    临床研究

  • ZHANG Ai-hua;ZHANG Dong-liang;YIN Dao-xin;DING Jia-xiang;DIAO Zong-li;CUI Wen-ying;ZHAO Wei;XU Rui;LIU Wen-hu.
    2012, 28(8): 587-591.
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    Objective To evaluate the effect of cardiovascular disease (CVD) on the healthcare costs associated with chronic kidney disease (CKD). Methods A total of 96 patients with stage 3-4 CKD treated at CKD clinic of Beijing Friendship Hospital, Capital Medical University were enrolled in the study. Their mean age was (61.6±9.5) years including 65 male and 31 female patients. A retrospective analysis of clinical material and health claims between January 2010 and October 2010 was conducted. Firstly, patients were grouped according to median CKD-associated healthcare cost and clinical characteristics were compared between two groups. Secondly, patients were stratified into three categories based on CVD prevalence (with acute cardiovascular events, with CVD but no acute events, and without CVD), and CKD-associated healthcare costs were assessed among the groups. Finally, the potential factors influencing CKD-associated healthcare costs were evaluated by optimal scaling regression analysis. Results During January to October in 2010, median CKD-associated healthcare costs was 13 960.5 yuan (interquartile range 10 226.5, 19 667.2 yuan). In the group with higher healthcare costs, more females, greater urine albumin-creatinine ratio, more emergency observations and/or hospitalizations caused by acute cardiovascular events, higher diabetes mellitus prevalence and calcium-phosphorus products, and lower eGFR and hemoglobin levels were found (P<0.05, respectively). In contrast, the total prevalence of CVD was not significantly different between the groups (P=0.386). When grouping by CVD prevalence, significant differences of CKD-associated healthcare costs were observed only between patients with acute cardiovascular events and the other two groups (P<0.01, respectively). The median healthcare cost of the former was approximately twice as higher as that of the other two groups, and the maximal cost was also found in the acute-cardiovascular-event group. For the optimal scaling regression analysis, both emergency observations and/or hospitalizations caused by acute cardiovascular events and diabetes mellitus entered the equation, and standardized coefficients were -0.538 and -0.217 respectively (P<0.01 and P<0.05). Conclusions Emergency observations and/or hospitalizations caused by acute cardiovascular events are important factors inducing high CKD-associated healthcare costs in patients with stage 3-4 CKD. Therefore, the prevention of acute cardiovascular events may be favorable to reduce CKD-associated healthcare costs. Larger and longer-time perspective studies are required to confirm it. In addition, diabetes mellitus also influences CKD-associated healthcare costs.
  • LIU Jing;MA Kun-ling;LIU Bi-cheng;GAO Min;ZHANG Xiao-liang;LIU Hong;WANG Yan-li.
    2012, 28(8): 592-596.
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    Objective To investigate whether low density lipoprotein receptor (LDLr) pathway involves in the progression of vascular calcification (VC) in hemodialysis patients under microinflammation. Methods Twenty-eight hemodialysis patients were divided into control and inflammation group according to plasma C-reactive protein level. Surgically removed tissues from radial artery of patients receiving arteriovenostomy were used in experiments. Foam cell formation and calcification deposition were observed by hematoxylin-eosin (HE) and alizarin red S staining respectively. VC-related protein expression, such as bone morphogenetic proteins-2 (BMP-2), collagen Ⅰ, alkaline phosphatase (ALP), and LDLr and its related nuclear factor of transcriptional regulation, such as sterol regulatory element binding protein-2 (SREBP-2) and SREBP cleavage-activating protein (SCAP), were detected by immunohistochemistry and immunofluorescence staining. Results HE and alizarin red S staining showed that there were parallel increased foam cell formation and calcium deposit in continuous cross-sections of radial arteries in inflammation group compared to control group, which were closely correlated with increased protein expressions of LDLr, SREBP-2, BMP-2, and collagen I as shown by immunohistochemical and immunofluorescent staining. Confocal microscopy confirmed that inflammation enhanced the translocation of SCAP/SREBP-2 complex from endoplasmic reticulum to Golgi, thereby activating LDLr gene transcription. Inflammation increased protein expression of ALP and reduced protein expression of alpha-smooth muscle actin, contributing to the phenotype conversion of vascular smooth muscle cells in calcified vessels from the fibroblastic to the osteogenic, which were the main cell components in VC. Further analysis showed that the disruption of LDLr pathway induced by inflammation was positively correlated with the enhanced expression of BMP-2 and collagen I(r=0.782, P<0.01; r=0.644, P<0.05). Conclusion Inflammation accelerates the progression of VC in hemodialysis patients through the disruption of LDLr feedback regulation.
  • LI Zhan-yuan;HUANG Wen;ZHENG Yu;YE Han-yang;JIN Ling-wei;YE Bai-ru;ZHOU Zhi-hong.
    2012, 28(8): 597-601.
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    Objective To examine the correlation of serum neutrophil gelatinase-associated lipocalin (NGAL) with dialysis adequacy, microinflammatory state and iron metabolism, and to assess the value of NGAL in identifying the dialysis adequacy in maintenance hemodialysis (MHD) patients. Methods A total of 150 MHD patients and 50 healthy control people were enrolled in the study. Clinical data were collected and serum NGAL, CRP, transferrin saturation, serum ferritin were measured. MHD patients were divided into adequacy group and non-adequacy group according to spKt/V value. Serum NGAL between two groups was compared and correlation of NGAL with spKt/V, inflammatory factors, and other clinical indexes was examined by Pearson, multivariate regression Logistic model and area under curve (AUC) of ROC. MHD patients were followed up for 3 months. Similar correlation analysis was carried out before and after follow up between two groups. Results Serum NGAL of MHD patients was higher compared to healthy people [(445.45±50.34) μg/L vs (50.02±6.45) μg/L]. Among 150 MHD patients, 95 were classified into adequacy group, while 55 into non-adequacy group according to spKt/V value. Serum NGAL was significantly higher in adequacy group as compared to non-adequacy group [(589.14±56.34) μg/L vs (360.13±46.23) μg/L, P<0.05]. Serum NGAL was positively correlated to spKt/V, CRP, TSAT [r=0.652, 0.825, 0.785, all P<0.05], but not to serum ferritin. Serum NGAL was also correlated to spKt/V, CRP, TSAT by multivariate regression Logistic model. AUC showed that NGAL level could reflex the condition of dialysis adequacy. During the follow up, all adequacy patients remained the same adequacy status. Thirty-eight out of 55 non-adequacy patients were qualified for complete adequacy after intervention. The serum NGAL before and after intervention was (368.14±56.21) μg/L and (360.56±46.23) μg/L respectively without significant difference. Conclusions Serum NGAL is obviously higher in MHD patients with dialysis adequacy compared to those without dialysis adequacy. Serum NGAL is positively correlated to spKt/V, CRP, TSAT. NGAL can reflex the condition of dialysis adequate in MHD patients.
  • LIN Xing-hui;YAN Yu-cheng;ZHU Ming-li;GU Le-yi;NI Zhao-hui;ZHANG Wei-ming;QIAN Jia-qi.
    2012, 28(8): 602-605.
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    Objective To investigate the clinical characteristics of twice-weekly hemodialysis patients. Methods Data were collected from Shanghai Renal Registry. A total of 1288 patients undergoing regular hemodialysis(HD) with dialysis adequacy index and other biochemical parameters in Shanghai in January 2007 were enrolled into the cohort study with 2 years follow-up. Clinical characteristics and outcome of twice-weekly HD patients were analyzed as compared with thrice-weekly HD patients. Results Compared with patients on thrice-weekly HD, the twice-weekly HD patients were significantly younger and had significantly shorter HD vintage, smaller body surface area, longer HD session time, higher single-pool Kt/V (spKt/V) and serum albumin but lower weekly Kt/V(P<0.05). There was no statistical difference in ultrafiltration volume between two groups. Kaplan-Meier survival analysis indicated that both groups had similar two-year survival. Multivariate Cox regression analysis showed that age, body mass index, serum albumin and weekly Kt/V were predictors of patient mortality. Conclusion It is acceptable for some hemodialys patients with twice-weekly HD, and close monitor of dialysis adequacy and volume status is necessary for this therapy model.
  • CHEN Zi-jin;CHEN Xiao-nong;MA Xiao-bo;HOU Liang;DING Bei;LING Hua-wei;LI Xiao;REN Hong;CHEN Nan.
    2012, 28(8): 606-610.
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    Objective To explore the incidence and associated factors of aortic artery calcification (AAC) by lateral lumbar X-ray score in maintenance hemodialysis (MHD) patients. Mehtods A total of 155 MHD patients with complete clinical data in our hospital were enrolled in the study. Lateral lumbar X-ray score of the abdominal aorta was used to determine AAC in MHD patients. Results Aortic calcification was most severe in front of the fourth lumbar segment and ameliorated in higher lumbar levels. 63.63% of MHD patients presented visible calcification in the abdominal aorta, and 28.39% had severe calcification with more than three segments. Age (OR=1.094, P<0.01), dialysis vintage (OR=1.013, P=0.022), triglyceride (OR=1.261, P=0.030) and phosphate level (OR=1.324, P=0.023) were risk factors of abdominal aorta calcification, however serum albumin level (OR=0.239, P=0.013) was protect factor of aortic calcification. Conclusions Incidence of AAC is quite high in MHD patients and associated with increasing of age, duration of hemodialysis, serum triglyceride, phosphate level and plasma albumin. The semi-quantitative X-ray method of determining vascular calcification is less expensive and may be widely available clinically.
  • 基础研究

  • CHENG Gen-yang;LI Hai-jian;LIU Zhang-suo; TANG Lin;QUAN Song-xia;
    2012, 28(8): 611-615.
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    Objective To investigate the effect of rosiglitazone on renal interstitial fibrosis in chronic cyclosporine nephropathy (CCN) rats. Methods Twenty-eight rats were randomly assigned to control group, rosiglitazone (RGZ, 5 mg&#8226;kg-1&#8226;d-1) group, cyclosporine A(CsA, 15 mg&#8226;kg-1&#8226;d-1) group, rosiglitazone(5 mg&#8226;kg-1&#8226;d-1) +CsA group. Real-time PCR and RT-PCR methods were used to investigate the expressions of OPN, RANTES on the 14th day and MMP-9, TIMP-1 on the 35th day in kidney of CCN respectively. Results In comparison with control group, the expressions of OPN, RANTES, MMP-9, TIMP-1 in CsA and RGZ+CsA groups were increased (P<0.05). In comparison with the CsA group , the expressions of OPN, RANTES, MMP-9, TIMP-1 in CsA+RGZ group significantly decreased (P<0.05). Conclusion Rosiglitazone may protect renal tissue after CCN by decreasing expressions of OPN, RANTES, MMP-9, TIPM-1.
  • TAN Ruo-yun;FANG Yi;SU Wei-fang;YANG Jun-wei;ZHANG Wei;GU Min.
    2012, 28(8): 616-621.
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    Objective To investigate the possible mechanism that hepatocyte growth factor (HGF) inhibits renal tubular epithelial-mesenchymal transition (EMT), and to determine whether Smurf2 expression induced by TGF-β1 can be reversed by HGF in normal rat kidney epithelial cells (NRK-52E). Methods Using rat NRK-52E cell line as an in vitro system, NRK-52E cells were incubated with 5 μg/L TGF-β1 for 0-24 h. Part of cells were pretreated with 20 μg/L HGF for 30 min or not, then incubated with or without 5 μg/L TGF-β1 for 1 h or 48 h. The other cells were transfected with pFlag-Smurf2 or Smurf2 siRNA for 24 h, then treated with or without 20 μg/L HGF for 24 h. The expressions of Smurf2, SnoN, E-cadherin, alpha-smooth muscle actin (α-SMA) and fibronectin (FN) were detected by Western blotting and indirect immunofluorescence staining assays. Results Compared to normal control, TGF-β1 could rapidly induce Smurf2 protein expression in a short time (P<0.01). Meanwhile, the expressions of FN and α-SMA were significantly induced, and the expression of E-cadherin was reduced in NRK-52E cells by TGF-β1. In contrast, in the NRK-52E cells pretreated with HGF, HGF could obviously inhibit Smurf2 expression induced by TGF-β1, and reversed the down-regulation of SnoN (P<0.01) and E-cadherin(P<0.05), the up-regulation of α-SMA (P<0.01) and FN (P<0.01) induced by TGF-β1. Moreover, overexpression of Smurf2 in NRK-52E cells could partly inhibit the up-regulation of SnoN protein by HGF, while down-regulation of Smurf2 could up-regulate the expression of SnoN induced by HGF. Conclusions HGF can abolish EMT induced by TGF-β1 in renal tubular epithelial cells through down-regulating Smurf2 expression and suppressing ubiquitin-proteasome dependent degradation of SnoN.
  • REN Zhi-long;LIANG Wei;DING Guo-hua;CHEN Cheng;ZHOU Min;XU Wan;YANG Hong-xia.
    2012, 28(8): 622-627.
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    Objective To evaluate the effect of angiotensinⅡ(AngⅡ) on the change of nephrin phosphorylation both in AngⅡ-infused rat model and cultured podocytes. Methods Thirty Wistar rats were subcutaneously embedded with osmotic minipumps and randomly divided into 3 groups according to receiving either AngⅡ at a dose of 400 ng&#8226;kg-1&#8226;min-1 or AngⅡ+ telmisartan at a dose of 3 mg&#8226;kg-1&#8226;d-1, or normal saline as a control group. Blood pressure and 24-hour urinary albumin were measured at 0 d, 7 d, 14 d, 21 d and 28 d of the experiment. Renal histomorphology was evaluated through electron microscopy. The concentrations of AngⅡ both in blood plasma and kidney were detected by radioimmunoassay. In vitro, cultured murine podocytes were exposed to Ang Ⅱ(10-6 mol/L) pretreated with or without losartan (10-5 mol/L) for different time periods. Nephrin and its phosphorylation expression were analyzed by Western blotting. The distribution of F-actin was presented by FITC-phallodin labeling. The change in phenomenon of F-actin was evaluated by cortical F-actin score index (CFS). Results (1)AngⅡ-infused rats exhibited increased AngⅡconcentration, significant hypertension and marked albuminuria. (2)In AngⅡ-infusion group, nephrin expression was decreased(P<0.05). AngⅡ-receiving rats displayed diminished phosphorylation of nephrin. (3)In vitro, the phosphorylation of nephrin was significantly reduced after AngⅡ stimulation for 3-6 hours(P<0.05). (4)AngⅡ stimulatation resulted in irregularly arrangement of F-actin followed by the redistribution of F-actin to podocyte periphery and formation of F-actin ring, in which the CFS obviously increased compared to control (P<0.05). Conclusions Phosphorylation of nephrin is important for the survival status of podocytes. AngⅡ-induced nephrin dephosphorylation may be an important molecular mechanism for AngⅡ-induced podocyte cytoskeleton rearrangement and foot process effacement.
  • BIAN Bao-ping;WAN Ying;CHEN Rong;ZHANG Chun-mei;TANG Hua;LUO Na;ZOU Ping.
    2012, 28(8): 628-632.
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    Objective To investigate the expression of periostin in the kidneys of rats with obstructive nephropathy and its relevance to renal interstitial fibrosis. Methods Eighteen male adult SD rats were randomly divided into sham group, model group and benazepril group (6 in each group). Unilateral ureteral obstruction (UUO) model was induced by ligating the left ureter of rats. RT-PCR was used to detect the mRNA expressions of periostin and TGF-β1, and ELISA to detect the protein expression of periostin, AngⅡ, and TGF-β1 in kidney tissue. Pathological changes of renal tissue were observed by HE and Masson staining. The protein expression of collagen Ⅰ(ColⅠ) in kidney tissue was examined by immunohistochemical staining. Results The mRNA expressions of periostin and TGF-β1 in model group increased markedly as compared with sham group (all P<0.05), and benazepril could decrease these mRNA expressions (all P<0.05). The protein expressions of periostin, AngⅡ and TGF-β1 in kidney tissue were significantly increased in model group as compared with sham group (all P<0.05), and benazepril could decrease these protein expressions (all P<0.05). The expression of periostin in kidney tissue was positively correlated with the expressions of AngⅡ,TGF-β1 and ColⅠ, as well as the degree of renal interstitial fibrosis (r=0.652, 0.781, 0.776 and 0.825 respectively, all P<0.05). Conclusion Periostin expression is up-regulated in kidney tissue of rats with obstructive nephropathy, which is associated to the over-deposition of extracellular matrix (ECM) in the kidneys of UUO rats.
  • TANG Lin;YI Run;YANG Bing;LI Hui;CHEN Hui-juan;WANG Liu-wei;LIU Zhang-suo.
    2012, 28(8): 633-638.
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    Objective To investigate the effect of valsartan on expression of HIF-1α, TIMP-1, MMP-9 in the process of renal interstitial fibrosis(RIF) in diabetic nephropathy(DN) rats. Methods Fifty-four SD rats were randomly divided to 3 groups: control group(group C), DN group (group D), valsartan treatment group (group T). Rats of group D and T were given streptozocin (STZ) by intraperitoneal injection to establish animal model of diabetes. After diabetic models were successfully made, rats of group T were treated by valsartan(40 mg&#8226;kg-1&#8226;d-1). Serum glucose, serum creatinine and urinary protein were measured at the 4th, 8th and 12th weeks. Masson staining was used to calculate the area of RIF. The methods of immunohistochemistry and real-time PCR were used to detect the expressions of HIF-1α, TIMP-1, MMP-9 in renal interstitium. Results Compared with group C, the areas of RIF were larger, 24-hour urinary protein, Scr were higher, the mRNA and protein expression of HIF-1α and TIMP-1 increased, while the mRNA and protein expression of MMP-9 decreased in D and T group(P<0.05). At the 8th, 12th weeks, compared with group D, the areas of RIF were smaller, 24-hour urinary protein, Scr were relatively lower expressions of HIF-1α and TIMP-1 decreased, while the eapressions of MMP-9 increased in group T (P<0.05). Conclusion Valsartan may delay the progress of RIF in DN rats by down-regulating the expressions of HIF-1α, TIMP-1, up-regulating the expression of MMP-9, then improve renal function.
  • TAN Zhi-cheng;YAN Yan;LI Rong-shan.
    2012, 28(8): 639-642.
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    Objective To observe the protective effect of different doses sodium hydrosulfide (NaHS) as a donor of hydrogen sulfide on renal ischemia reperfusion injury (IRI). Methods Twenty-eight healthy male Wistar rats were randomly divided into 4 groups, namely sham operation (Sham) group (n=7), renal ischemia reperfusion(IR) group(n=7), sodium hydrosulfide (NaHS) high dose group(n=7), sodium hydrosulfide low dose group (n=7). After excision of the right kidney, two dose NaHS group(300 nmol/min, 1.5 μmol/min) received 15-minute continuous administration via left renal artery. Sham group and IR group received same volume of saline. Five minutes after stopping drug, the left renal pedicle in NaHS group and IR group was clipped with no damage arteriole occlusion. After 45 minutes blockade, a model of acute renal ischemia reperfusion injury was established. Sham group underwent the same procedure as model groups without clamping the left renal artery. Specimens of renal tissue and blood were harvested at 24-hour after blood flow restore in the kidney. BUN and Scr were measured. Kidney pathological damage was semi-quantitatively analyzed. Production of H2S in renal tissue was detected. Expression of cystathionine γ lyase (CSE) and cystathionine β synthase (CBS) mRNA in kidney tissue was examined by real-time PCR. Results Compared with sham group, BUN and Scr increased significantly (P<0.01), kidney tissue H2S production decreased significantly (P<0.01), mRNA expression of CSE and CBS down-regulated significantly (P<0.01) in IR group. The kidney pathology of sham group was normal, while acute tubular necrosis was found in IR group. Compared with IR group, BUN and Scr decreased significantly (P<0.01), kidney tissue H2S production increased significantly (P<0.05), mRNA expression of CSE and CBS up-regulated significantly (P<0.01) in NaHS groups. Pathological damage of acute tubular necrosis was significantly improved in NaHS pretreatment group. There was no significant difference between two NaHS doses groups. Conclusion Hydrogen sulfide has a protective effect on renal IRI.