Objective To explore the effect of FTY720 on human hepatocellular carcinoma cell line HepG-2 of invasion ability. Methods HepG-2 cells were cultured,and divided into 4 groups,namely control and FTY720 (0.05 g/ml,0.10 g/ml,0.15 g/ml) groups.The cell invasion ability was observed by the cell invasion assay.mRNA expression of MMP-2,MMP-9 was measured by RT-PCR.MMP-2,MMP-9 protein expression was measured by ELISA method.Results The invasive ability of cancer cells in each group significantly weaken,with the FTY720 concentration increased.The number of control group was (110±3)(P<0.05),the number of O.05 g/ml FRY720 treatment group cells was (74±4),while the number of cells of O.10 g/ml,0.15 g/ml the drug treated group,Was (42±3),(25±5)(P<0.01).Compared with control group,the MMP-2,MMP-9 expression of different concentrations of FTY720 groups decreased significantly (P<0.05).As the expression of FTY720 concentration increased,the MMP-2,MMP-9 expression decreased (P<0.01).Compared with control group,the MMP-2,MMP-9 protein expression of different concentrations of FTY720 groups decreased significantly (P<0.05), but the protein expression and concentration of FTY720 is inversely proportional in a certain range. Conclusion The invasive ability of HepG-2 cells can be inhibited by FTY720,and with FTY720 concentration increased,the invasive ability decreased gradually.One of the mechanisms may be related to reduce the MMP-2,MMP-9 protein and gene expression.

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.

【Objective】To observe the anti-depression effect of Shugan Hewei Granules(SHG) on mice with behavioral despair.【Methods】Kunming mice were randomized into 5 groups: model group,low-,moderate-and high-dose SHG groups(in the dosages of 0.25,0.5 and 1.0 g?kg-1?d-1respectively) and chloropromazine(CP) group.Tail suspension test and forced swimming test were used to investigate immobility time of mice.【Results】Moderate-and high-dose SHG shortened the immobility time obviously during tail suspension test and forced swimming test(P

reression of M 2 receptor mRNA of rat myocardium intoxicated by soman under high altitude hypoxia. Methods The expressions of M 2 receptor mRNA under hypoxia, soman intoxication and soman intoxication under hypoxia were detected by semi quantitative RT PCR, respectively. Results The expression of M 2 receptor mRNA increased in the high altitude hypoxia group. Both simple soman intoxication and combined soman intoxication and hypoxia decreased the expression of M 2 receptor rapidly. But under hypoxia, the expression increased significantly at 12 h and 24 h. Conclusion M 2 receptor was sensitive to nerve agents. Compared to simple soman intoxication group, the expression of M 2 receptor increased in combined soman intoxication and hypoxia group. This may be one of the major factors leading to aggravation of the injury of heart function by nerve agents in high altitude area.

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.

Objective　To observe the changes of the concentration of Ca2+, contents of cAMP, CaM and activity of Ca2+/CaM-PK II in pheochromocytoma PC12 cells after combined soman and hypoxia injury. Methods　The changes of ［Ca2+］, and activity of CaM, cAMP and Ca2+/CaM-PK II in PC12 cells were studied after combined soman and hypoxia injury with radioimmunoassay. Results　The changes of ［Ca2+］, the contents of CaM, cAMP were significantly higher in hypoxic and soman intoxicated group than in soman intoxicated group and control group under hypoxia; but the activity of Ca2+/CaM-PK Ⅱ were significantly decreased. Conclusion　［Ca2+］, CaM, cAMP and Ca2+/CaM-PK Ⅱ exert important role in the damage of PC12 after combined soman and hypoxia injury.

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.

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.

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 .

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.