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Objective To observe the effect of aralia saponins on the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in the kidney of diabetic nephropathy mice.Methods After 10 days of adaptive feeding,90 clean Kunming mice were randomly divided into the normal group (n =10) and 5 model groups (model group,positive drug benazepril intervention group,aralia saponins low,middle and high doses treatment groups).Excepted the normal group,the kidney damage model of type 2 diabetes mellitus in mice was induced by high-fat and high-sugar diet for one month plus disposable streptozotocin (STZ).The model was successfully constructed and killed after 6 weeks of treatment.A total of 25 mice failed to establish the model.And totally 55 mice were randomly divided into 5 groups with 11 mice in each group.The serum changes of blood urea nitrogen (BUN),serum creatinine (SCr),insulin,inflammatory factors interleukin-1α (IL-1α),interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in each group were detected.The expression of vascular endothelial growth factor and MMP-9 protein in renal tissue were detected by immunohistochemistry and Western blot.Results Compared with the normal group,the levels of blood glucose,insulin,BUN,SCr,IL-1α,IL-6 and TNF-α in the model group were significantly increased (P ＜ 0.05).The above indexes were decreased in positive drug group and aralia saponins treatment groups.The contents of insulin,BUN and SCr in the high dose of aralia saponins group were significantly lower than those parameters in benazepril group (P ＜ 0.05).In addition,the contents of blood glucose,IL-1α,IL-6 and TNF-α in the three dose aralia saponins groups were significantly lower than those parameters in the benazepril group (P ＜ 0.05).Compared with the normal group,the expression level of VEGF protein in the model group was significantly higher (P ＜ 0.05),and the expression level of MMP-9 protein was significantly lower (P ＜ 0.05).Compared with the model group,both benazepril and aralia saponins can reduce VEGF (P ＜ 0.05),increase MMP-9 (P ＜ 0.05).In addition for VEGF and MMP-9,the high dose of aralia saponins group and benazepril group was basically same.Conclusions Aralia saponins can significantly reduce blood glucose,insulin and serum inflammatory factors,while downregnlate VEGF and increase MMP-9 protein levels,thereby protecting the kidneys of diabetic nephropathy mice.
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Objective To study the effect of aralia total saponins on renal function of type 2 diabetic mice, and its effect on the Bax and Bcl-2 protein in renal tissues, in order to provide some references for the development of aralia total saponins. Methods The mice were divided into the normal group, model group, positive control group, low, medium and high dose aralia total saponins group by random number method. Except the normal group, the others were received with high-fat diet for one month+one-time large dose of streptozotocin (STZ) to induce type 2 diabetic model, and then the mice in the normal group and the model group were intragastrically administered with the same volume of normal saline, and the mice in the positive control group was given 1 mg/kg of benazepril solution, and the low, medium and high dose groups were given 30, 60, 120 mg/kg aralia total saponins. The body weight of 1 ml/kg mice was intragastrically administered once a day. After treatment for 6 weeks they were sacrificed, and the serum insulin, and SOD and MDA levels were measured, the urine creatinine (Cr), urea nitrogen (UN), and uric acid (UA) levels were also measured. The immunohistochemistry and Western blot were used to detect the Bax and Bcl-2 protein expression in kidney tissues. Results Compared with the model group, the blood glucose and insulin resistance index in the low, medium and high doses aralia total saponins group were significantly decreased (P<0.05); the levels of urine UN, Cr and UA significantly decreased (P<0.05); The serum SOD level increased and the MDA level significantly decreased (P<0.05). The average gray value of Bcl-2 increased (92.26 ± 11.36, 107.17 ± 9.26, 132.65 ± 8.45 vs. 56.42 ± 16.24) in kidney tissue. The average gray value of Bax (152.62 ± 9.86, 124.48 ± 10.36, 92.29 ± 10.10 vs. 171.38 ± 15.18) significantly decreased ( P<0.05); Bax protein (0.81 ± 0.06, 0.75 ± 0.07, 0.52 ± 0.09 vs. 2.02 ± 0.09) significantly decreased, but Bcl-2 protein (0.92 ± 0.08, 0.94 ± 0.12, 1.27 ± 0.07 vs. 0.30 ± 0.09) significantly increased (P<0.05). Conclusions The aralia total saponins can reduce blood sugar levels, meanwhile improve renal function in type 2 diabetic mice. The mechanism may be may be that aralia total saponins could improve the antioxidant capacity and inhibition of renal cell apoptosis.
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Objective To study the therapeutic effect of bitter gourd saponins on salt-sensitive kidney injury induced by high salt diet and its possible mechanism.Methods 50 Sprague Dawley (SD) rats were randomly divided into normal group,model group and low-dose,middle-dose and high-dose treatment group after 10 days of adaptive feeding.Each group had 10 rats.Except the normal group,the other four groups were given high salt diet (4.0％ high salt diet) to induce salt-sensitive kidney damage in rats.The normal group and the model group were given 1.0 m/(kg · d) normal saline,and the three dosage groups of total saponins of balsam pear were given 10 mg/(kg · d),20 mg/(kg · d) and 40 mg/(kg · d) respectively.After 8 weeks of treatment,rats were sacrificed and collect the 24-hour proteinuria,creatinine.Serum creatinine,serum aldosterone,serum sodium and serum potassium were measured,and renal histopathology and the expression of podocin and nephrin were detected.Results Pathological examination of model group showed obvious glomerular sclerosis and renal interstitial fibrosis,and glomerular sclerosis in the treatment group was obviously improved by bitter gourd saponins;The systolic pressure in the model group was 170 mmHg,significantly higher than that of the normal and treatment groups,the systolic blood pressure of the treatment groups were obvious decreased when treated by bitter gourd saponins (P ＜ 0.05);Compared with normal group,serum creatinine and 24 h proteinuria / urine creatinine in model group were significantly increased (P ＜ 0.05),while creatinine clearance rate and aldosterone were significantly decreased (P ＜ 0.05),and the above indexes in bitter gourd saponins treatment group were significantly improved;Compared with the model group,the protein and mRNA expression of podocin and nephrin were significantly decreased (P ＜ 0.05),while the two indexes can be revered by bitter gourd saponins in treatment group (P ＜ 0.05).Conclusions The bitter gourd saponins can significantly improve the symptoms of salt-induced hypertensive nephropathy in rats,which may be related with the expression of podocin and nephrin in renal tissue,thereby inhibiting glomerulosclerosis and improving renal interstitial fibrosis.
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Objective To observe the expression of interferon induced protein (IP)-10 and the role of nuclear factor-kappaB (NF-κB) signaling pathway in rat peritoneal mesothelial cells (RPMCs) under the action of lipopolysaccharide (LPS).Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and cultured under defined in vitro conditions.The cells were exposed respectively to different concentrations of LPS (0,10,100,1 000,10 000 ng/ml) for 3 h or treated with LPS (100 ng/ml)for different time points (0,1,3,6,12,24,48 h).For observing the effect of LPS on the expression of p-p65 and p65,the RPMCs were treated with LPS (100 ng/ml) for different time points (0,15,30,60,120 min).For observing the effect of BAY11-7085 on the expression of IP-10 mRNA,the RPMCs were treated by LPS or pretreated with BAY11-7085 (5 μ mol/L) for2 h,then treated with LPS for another 3 h,respectively.Expression of IP-10 mRNA was examined by reverse transcription-polymerase chain reaction (RT-PCR).Expression of NF-κB and p-NF-κB protein was detected by Western blot.The secretion of IP-10 was determined by enzyme-linked immunosorbent assay (ELISA).Results Compared with the control group,stimulation of RPMCs with 10 ng/ml LPS resulted in a significant increase in the expression of IP-10 mRNA (P ＜0.05).1 000 ng/ml LPS has the strongest effect on IP-10 expression compared with that of 10 ng/ml and 100 ng/ml LPS.Treatment with 100 ng/ml LPS resuhed in time-dependent increase in the gene level of IP-10,with the peak at 3 h.However,after that time point,the gene level of them was gradually attenuated.Following treatment with LPS (100 ng/ml),the level of p-NF-κB began to increase at 15 min,gradually reached the peak at 1 hour,and then decreased.But the level of which at 2 h is still significant higher than that of medium control.5 μmol/L BAY11-7085 significantly decreased the up-regulation of IP-10 induced by LPS.Conclusions LPS enhanced the expression of IP-10 on RPMCs in a concentration-dependent and a time-dependent manner.LPS induced expression of IP-10 depended on the NF-κB signal transduction pathway.
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Objective To investigate the protective effect of aralia total saponins on renal injury induced by streptozotocin (STZ) in type 2 diabetic mice, meanwhile to explore its protective mechanism. Methods Fifty male Kunming mice were randomly divided into the normal group, the model group and the aralia total saponins low, middle and high does groups. All the rats were given high fat diet 8 weeks and then received STZ 45 mg/kg to built type 2 diabetic mice model, except the noraml group. After the models establishment,the aralia total saponins low, middle and high does groups were given the aralia total saponins 30, 60, 120 mg/kg treatment, andthe normal group and the model group were given the equal normal saline, once each day. After 4th and 8th week administration, the urinary protein levels of 24 h in each group were detected. After the last treatment, all the mice were sacrificed to detected the changes of blood glucose, insulin and inflammatory related factors. Immunohistochemistry, Western blotting and real-time quantitative PCR were used to observe the expression of TLR2 and TLR2 in the kidney tissue. Results Compared with the model group, the low, middle and high does groups in 24-hour proteinuria, blood glucose, insulin resistance index decreased (P<0.05), the insulin increased(P<0.05). The serum TNF-α (16.66 ± 0.20 ng/L, 14.49 ± 0.27 ng/L, 13.52 ± 0.22 ng/L vs.20.33 ± 0.56 ng/L),IL-1β(0.46 ± 0.04 ng/L,0.44 ± 0.04 ng/L,0.37 ± 0.04 ng/L vs.0.55 ± 0.05 ng/L),NF-κB (28.71 ± 6.14 ng/L, 26.26 ± 5.48 ng/L, 25.69 ± 5.61 ng/L vs. 36.55 ± 8.90 ng/L) significantly decreased (P<0.05).The kidney TLR2 mRNA(1.92 ± 0.18,1.46 ± 0.23,1.28 ± 0.21 vs.2.69 ± 0.22),TLR4 mRNA(2.20 ± 0.19,2.08 ± 0.27,1.57 ± 0.22 vs.2.78 ± 0.23),TLR2 porteins(0.82 ± 0.11,0.52 ± 0.06,0.44 ± 0.07 vs.0.77 ± 0.13),TLR4 proteins(0.52 ± 0.04,0.42 ± 0.09,0.26 ± 0.06 vs.0.86 ± 0.12)significantly decreased(P<0.05). Conclusions The aralia total saponins can significantly reduce the blood glucose, insulin resistance index and 24-hour urinary protein in type 2 diabetic nephropathy of mice, increase the insulin, and analyzing its mechanism may be total saponins can inhibit the expression of TLR2 and TLR4 protein in the kidney, and further reduce the inflammatory response.
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Objective To investigate the effect of bitter gourd saponins on the salt -sensitive hypertension caused kidney damage in rats ,and analyze the mechanism of its therapeutic effect .Methods 50 SD rats were fed for 10 days with normal diet ,and then based on random number method ,the rats were randomly divided into 5 groups:the normal control group,model control group and the low total saponins bitter,medium and high dose treatment group,10 rats in each group.Then,the rats in the normal control group and the model control group were given 1mL· kg-1 · d-1 normal saline,and the rats in bitter gourd saponins groups were given high salt diet 8 weeks to establish the model .After 4 weeks,the total saponins of bitter gourd treatment groups were given 10,20 and 40mg· kg-1· d-1.8 weeks later, the rats were sacrificed and the renal pathology was detected by HE staining .The changes of blood pressure , heart rate,urinary function and blood renal function were also analyzed .At last,Western blot and semi -quantitative PCR were used to detect the endothelin -1 (ET-1) in renal tissue.Results For the systolic blood pressure,in whole of treatment,the normal control group maintained at about 120 mmHg, while the model control group maintained at 170mmHg,which of the bitter gourd saponins three doses groups compared to the model control group was significantly lower (t=1.765,1.982,2.126,all P<0.05),further improved.And the heart rate had no statistically significant difference among the groups(all P>0.05).Compared with the normal control group,the blood urea nitrogen (BUN), creatinine (Cr),uric acid (UA),urine volume,urinary protein and N -acetyl beta -D-Glucosaminidase (NAG) levels in the model control group were significantly increased (t=28.703,33.932,29.298,4.695,10.989,10.871, all P<0.05),which in the bitter gourd saponins treatment groups were significantly decreased (all P<0.05).HE staining showed that the rats in the normal group were normal ,and the model control group had obvious glomerular sclerosis and renal interstitial fibrosis , after bitter gourd saponins treatment , the condition was significantly improved .Immu-nohistochemistry,Western blot and semi-quantitative PCR showed that normal control group had almost a little expression of ET-1,and compared with the normal control group ,which of the model control group was significantly increased ( t=14.650,11.387,all P<0.05),and the ET-1 expression in the bitter gourd saponins treatment groups significantly decreased.Conclusion The bitter gourd saponins can significantly improve the symptoms of hypertension and renal damage induced by high salt diet in rats ,which may be related with regulation of ET -1 expression in renal tissue .
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Objective To evaluate the effect of isosceles right angle triangulation on deep vein puncture under ultrasound localization. Methods In this prospective randomized case-controlled clinical study, the enrolled patients who required deep veins puncture to establish transient hemodialysis pathway were randomly divided into treatment group treated with ultrasound localization puncture based on isosceles right triangle theory, and control group treated with traditional localization puncture following exploration of blood vessels using color doppler ultrasound, with 46 cases in each group. The differences in gender, age, causes for hemodialysis, puncture sites, successful rate of initial puncture, rates of complications and cathetering-consuming time were compared between two groups. Results The differences in gender, age, causes for hemodialysis and puncture sites between two groups were not statistically significant. In control group, 30 patients received puncture through internal jugular veins, among whom 16 received successful initial puncture, with the successful rate of initial puncture of 53.33%, and the cathetering-consuming time of (545.5 ± 120.3) s; and 16 patients received puncture through femoral veins, among whom 8 received successful initial puncture, with the successful rate of initial puncture of 50.00%, and the cathetering-consuming time of (500.6 ± 120.6) s. Meanwhile, in control group, 3 patients had hematoma in puncture sites, and another 3 had arterial mis-puncture, with the total rate of complications of 13.04%(6/46). In treatment group, 32 patients received puncture through internal jugular veins, among whom 31 received successful initial puncture, with the successful rate of initial puncture of 96.88%, and the cathetering-consuming time of( 99.5 ± 34.8) s;and 14 patients received puncture through femoral veins, among whom 14 succeed in initial puncture, with the successful rate of initial puncture of 14/14, and the cathetering-consuming time of (99.8 ± 44.8) s. None in treatment group had hematoma in puncture sites or arterial mis-puncture. Both groups finished deep veins puncture intubation successfully. Treatment group was markedly higher in the successful rate of initial puncture (P<0.05), notably shorter in cathetering-consuming time (P<0.05), and prominently lower in the total rate of complications than control group (P<0.05), and all differences were statistically significant. Conclusions Ultrasound localization based on isosceles right triangle theory has superiority to traditional B-ultrasound localization in deep veins puncture in that it can promote the successful rate of initial puncture and shorten cathetering-consuming time without increasing the occurrence rates of complications, and can be completely operated by single clinical physician independently, thus deserving to be popularized in clinical practice.
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Objective To evaluate the effect of isosceles right angle triangulation on deep vein puncture under ultrasound localization. Methods In this prospective randomized case-controlled clinical study, the enrolled patients who required deep veins puncture to establish transient hemodialysis pathway were randomly divided into treatment group treated with ultrasound localization puncture based on isosceles right triangle theory, and control group treated with traditional localization puncture following exploration of blood vessels using color doppler ultrasound, with 46 cases in each group. The differences in gender, age, causes for hemodialysis, puncture sites, successful rate of initial puncture, rates of complications and cathetering-consuming time were compared between two groups. Results The differences in gender, age, causes for hemodialysis and puncture sites between two groups were not statistically significant. In control group, 30 patients received puncture through internal jugular veins, among whom 16 received successful initial puncture, with the successful rate of initial puncture of 53.33%, and the cathetering-consuming time of (545.5 ± 120.3) s; and 16 patients received puncture through femoral veins, among whom 8 received successful initial puncture, with the successful rate of initial puncture of 50.00%, and the cathetering-consuming time of (500.6 ± 120.6) s. Meanwhile, in control group, 3 patients had hematoma in puncture sites, and another 3 had arterial mis-puncture, with the total rate of complications of 13.04%(6/46). In treatment group, 32 patients received puncture through internal jugular veins, among whom 31 received successful initial puncture, with the successful rate of initial puncture of 96.88%, and the cathetering-consuming time of( 99.5 ± 34.8) s;and 14 patients received puncture through femoral veins, among whom 14 succeed in initial puncture, with the successful rate of initial puncture of 14/14, and the cathetering-consuming time of (99.8 ± 44.8) s. None in treatment group had hematoma in puncture sites or arterial mis-puncture. Both groups finished deep veins puncture intubation successfully. Treatment group was markedly higher in the successful rate of initial puncture (P<0.05), notably shorter in cathetering-consuming time (P<0.05), and prominently lower in the total rate of complications than control group (P<0.05), and all differences were statistically significant. Conclusions Ultrasound localization based on isosceles right triangle theory has superiority to traditional B-ultrasound localization in deep veins puncture in that it can promote the successful rate of initial puncture and shorten cathetering-consuming time without increasing the occurrence rates of complications, and can be completely operated by single clinical physician independently, thus deserving to be popularized in clinical practice.
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Objective To study the influence of hemodiafiltration on high-sensitivity C-reactive protein (hs-CRP),serum parathyroid hormone (iPTH) and cardiac function in uremic patients undergoing maintenance hemodialysis (MHD).Methods Forty patients with uremia were divided into hemodiafiltration (HDF) group (n =20) and hemodialysis(HD) group(n =20) by random numbers.Serum hs-CRP and iPTH were measured before treatment and at 6 months after treatment.Their left ventricular diastolic diameter (LVEDD),left ventricular end-systolic dimension (LVESD),interventricular septal thickness (IVST) and left ventricular ejection fraction(LVEF) were measured by echocardiogram.Ten healthy volunteers were recruited as the normal controls.Results After treatment,serum hs-CRP and iPTH decreased significantly in the HDF group (hs-CRP:(3.32±1.t4) mg/L vs.(7.84 ±2.21) mg/L,t =2.072,P =0.046;iPTH:(155.36 ±81.33) ng/L vs.(190.25 ±82.31) ng/L,t =2.121,P =0.023).No significant difference of serum hs-CRP and iPTH was observed in the HD group before and after treatment (hs-CRP:(7.91 ±2.11) mg/L vs.(7.68 ± 1.86) mg/L,t =1.731,P =0.111 ; iPTH:(177.43 ± 85.43) ng/L vs.(184.35 ± 87.21) ng/L,t =1.331,P =0.210).Color Doppler indices of cardiac structure of the HDF group significantly improved after treatment compared with before treatment (LVEDd:(55.3 ±3.7) mm vs.(58.8 ±4.3) mm,t =2.345,P =0.015 ;LVESd:(33.5 ±2.6) mm vs.(36.4±4.3) mm,t=2.178,P=0.046;IVST:(10.2±1.7) mmvs.(13.1 ±1.8) mm,t=2.630,P=0.012;LVEF:(55.3 ±2.6)％ vs.(42.5 ±3.3)％,t =2.860,P =0.010;LVMI:(132.5 ±9.1)g/m2 vs.(137.4 ± 8.7) g/m2,t =2.871,P =0.009).After treatment,cardiac structure of the HDF group was significantly better than that of the HD group (LVEDd:(55.3 ± 3.7) mm vs.(59.1 ± 4.2) mm,LVESd:(33.5 ±2.6) mm vs.(36.1 ±3.4) mm,IVST:(10.2±1.7) vs.(12.4 ±1.3) mm,LVEF:(55.3 ±2.6)％vs.(43.4±2.7)％,LVMI:(132.5 ±9.1) g/m2 vs.(139.4 ±8.9) g/m2;P ＜0.05).During the six month treatment,the incidence rate of cardiovascular events of the HDF group was significantly less than the HD group (36.39％(524/1440) vs.(72.08(1038/1440),x2 =21.583,P＜0.01).Conclusion HDF may efficiently remove iPTH,improve cardiac function of uremic patients,and help them recover from micro-inflammatory state.
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Objecfive 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 O h(control),12 h,24 h,48 h,72 h after sefrum-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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Objective To investigate the effects of hemodialysis (HD) and peritoneal dialysis (PD) on the complications and outcomes after renal transplantation. Methods Clinical data of 402 renal transplant recipients maintained on dialysis for more than 3 months were retrospectively studied and divided into 2 groups: HD group(n=303)and PD group(n=99). Among them, 345 recipients were followed up for an average of (30.2±15.2) months. The impact of HD and PD on the acute rejection, delayed graft function (DGF), infection, chronic rejection and the graft and patient survival rates were analyzed. Results The mean dialysis duration was significantly longer in PD group and the hepatitis B infection rate was significantly higher in HD group. There were no signiticant differences between the HD and PD groups in regarding to primary disease for end-stage renal disease, age, gender, blood pressure, hemoglobin, HLA match, hot and cold ischemia time, and hepatitis C vires infection. The incidence of DGF, acute and chronic rejection, and cytomegalovirus and other infections between HD and PD groups were not significantly different. However, the graft loss happened more frequently in hepatatis B patients than that in non hepatitis B patients (19.23% vs 8.86%, P=0.021), and the post-transplant infection ocurred less in non hepatits B patients with PD. The acute rejection episodes were higher in HD patients who received pretransplant dialysis for more than 12 months (P<0.05). The overall recipients survival rates of HD and PD groups were similar (1-year: HD 94.34%, PD 91.25%;5-year: HD 92.83%, PD 90%), and the same as the graft survival rates in HD and PD groups (1-year: HD 93.21%, PD 96.25%;5-year: HD 87.17%, PD 91.25%). Conclusions The influences of PD and HD on the complications after renal transplantaton, 1-year and S-year recipients and graft survival rates are similar, so both HD and PD can be chosen as the pretransplant dialysis modality. As the incidence of acute rejection increases with time in HD, it is better to shorten the time of pretransplant dialysis to decrease the complication.