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Objective:To explore the potential of Taenia solium (Ts) 14-3-3.3 protein as a candidate molecule for cysticercosis vaccine. Methods:Sixty Kunming mice with the body weight of 18 - 22 g were selected and divided into 3 groups according to their body weight via the random number table method, including normal saline control group (control group), Ts14-3-3.3 recombinant protein vaccine group (vaccine group), and Ts14-3-3.3 recombinant protein vaccine + adjuvant group (vaccine + adjuvant group), with 20 mice in each group. The multi-point subcutaneous injection method was adopted. After the first immunization at 0 week, the booster immunization was carried out twice, a total of 3 times, with an interval of 2 weeks. Four mice in the three groups were killed at 0, 2, 4, 6 and 8 weeks after the first immunization, and the blood of eyeballs and spleen were collected aseptically for serum separation and preparation of spleen lymphocytes suspension [treatment: cell suspension, antigen-stimulate and concanavalin (Con) A-stimulate], respectively. The levels of mouse serum specific immunoglobulin (Ig) G, IgG2a, IgG1 and IgE were detected by indirect enzyme-linked immunosorbent assay (ELISA). The proliferation level of mouse spleen lymphocytes was detected via the CCK-8 method. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-12, IL-13 and IL-10 in culture supernatant of mouse spleen lymphocytes were determined by double-antibody sandwich ELISA.Results:The IgG, IgG2a, and IgG1 levels of the vaccine and vaccine + adjuvant groups immunized for 2 to 8 weeks were higher than those of the control group, and the above indicators of the vaccine + adjuvant group were higher than those of the vaccine group ( P < 0.05). With the same treatment between the groups, the proliferation levels of spleen lymphocytes and the levels of TNF-α, IL-12, IL-13, and IL-10 in the culture supernatant after 2 - 8 weeks of immunization were statistically significantly different ( P < 0.05); the proliferation levels of spleen lymphocytes and the levels of TNF-α, IL-12, IL-13 and IL-10 in the culture supernatant of the vaccine and vaccine + adjuvant groups immunized for 2 to 8 weeks were higher than those of the control group, and the above indicators of the vaccine + adjuvant group were higher than those of the vaccine group ( P < 0.05). When treatment was different in the group, the proliferation levels of spleen lymphocytes and the levels of TNF-α, IL-12, IL-13 and IL-10 in the culture supernatant of the antigen-stimulate and ConA-stimulate were higher than those of the cell suspension, and the above indicators of the ConA-stimulate were higher than those of the antigen-stimulate ( P < 0.05). Conclusion:The recombinant protein vaccine of Ts14-3-3.3 can induce an effective immune response in mice.
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To explore the relationship between serum 1,25(OH)2D3 and oxidative stress in patients with diabetic nephropathy (DN).The serum levels of 1,25(OH)2D3 in DN patients were significantly lower than those in DM and healthy controls.And the 1,25(OH)2D3 level was negatively correlated with advanced oxidation protein products ( AOPP ) , reactive oxygen species ( ROS ) and positively correlated with superoxide dismutase (SOD) in DN patients.1,25(OH)2D3, low density lipoprotein-cholesterol (LDL-C), body mass index ( BMI) , serum creatinine and age were associated with the level of oxidative stress in DN patients.With the progression of CKD , the serum levels of AOPP and ROS increased significantly while those of SOD and 1,25(OH)2D3 decreased markedly.
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Objective To estimate dry weight (DW) and prevent dialysis-related hypotension and hypertension with the on-line monitoring of relative blood volume (RBV) and other judgments.Methods One hundred and eight maintenance hemodialysis patients were assigned to three groups according to their blood pressure:normal blood pressure group (A group,n=43),hypotension group (B group,n=35) and hypertension group (C group,n=35).The level of hemoglobin,serum albumin,dialysis adequacy were determined.Systolic blood pressure,diastolic blood pressure,mean arterial pressure,heart rate,ultrafiltration volume,relative blood volume changes and the corresponding clinical symptoms were monitored during hemodialysis in all patients.Each of the patients was continuously monitored of the indicators above for 10-12 times.At the observing period,the inferior vena cava diameter (IVCD),brain natriuretic peptide (BNP) and cardiothoracic ratio(CTR) were measured.Then according to the monitoring results,appropriate clinical interventions were given under on-line blood volume monitoring guidance.Results (1)The shape of RBV curve in group A showed doubleexponential curve early,then down to the final linear decling ended during hemodialysis.(2)The RBV curve in group B was stable in the former two hours,then rapidly linear declined.RBV changes were significantly higher in group B than group A (P ＜ 0.05),but when changes in RBV were plotted against ultrafiltration volume,there was no significant difference in the two groups.The level of RBV reduction at which symptomatic hypotension occurred showed considerable inter-individual variability (P ＜ 0.05,coefficient of variation=0.28).(3)The RBV curve in group C slowly linear declined.At the end of dialysis,RBV changes were significantly lower in group C than group A (P ＜ 0.05).(4)The IVCD values in three groups of patients before dialysis were greater than normal,significantly decreased after the dialysis (P ＜ 0.05),but that in group B and group C were still greater than that in group A (P ＜ 0.05).The BNP values were significantly greater in three groups before and after dialysis (P ＜ 0.05),but after dialysis,the values decreased significantly than that before dialysis (P ＜ 0.05).(5)After appropriate clinical intervention were given under on-line blood volume monitoring in hemodialysis,the patients of group B controlled weight gain,and even cut dry weight,the RBV change significantly decreased at the end of dialysis and significantly reduced the incidence of hypotension events (P ＜ 0.05); When the patients of group C cut dry weight,increased ultrafiltration,the RBV change increased,the mean arterial pressure decreased significantly than before (P＜ 0.05).Conclusions (1)Hemodialysis patients with symptomatic hypotension show larger RBV decline rate in the forth hour and lager total RBV changes,which provides important information for forecasting the symptomatic hypotension in hemodialysis.(2)IVCD and CTR have certain significance to the adjustment of dry weight,but the BNP has guiding significance to volume change.(3)On-line monitoring of RBV can effectively guide the adjustment of dry weight,reduction of symptomatic hypotension occruence,and controlling of refractory hypertension in hemodialysis.
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Objective To explore the effect of suramin on the epithelial-mesenchymal transition (EMT) and the excretion of transforming growth factor-β1 (TGF-β1) in peritoneal mesothelial cells (PMCs) induced by high concentrations of glucose solution (GS).Methods Cultured PMCs were divided into three groups:(1) normal control group; (2) GS-treated group:cells were treated with 1.5％,2.5％,4.25％ GS for 12 h,24 h,48 h,respectively; (3) Suramin-treated group:PMCs cultured with 4.25％ GS were exposed to different doses of suramin (25,50,100 μmol/L) for 48 h.Expression levels of α-smooth muscle actin (α-SMA) and E-cadherin were detected by Western blotting and the concentration of TGF-β1 in the culture supernatant was determined by ELISA.Results Compared with normal control group,GS-treated PMCs exhibited a time-dependent increase in the expression of α-SMA,and decrease in the expression of E-cadherin.GS also stimulated PMCs to secrete TGF-β1.In the presence of suramin,GS-induced α-SMA expression and TGF-β1 production were reduced,E-cadherin expression was increased.Conclusions Suramin can inhibit high glucose-induced EMT of PMCs by down-regulating the expression of TGF-β1.Suramin may be a novel therapeutic agent for the treatment of peritoneal fibrosis.
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Objective To investigate the prevalence and risk factors of chronic kidney disease (CKD) in middle-aged and older adult population in Shanghai Pudong New District.Methods Two thousand residents aged 45 years and above were randomly selected for questionnaire survey and health check-up in Shanghai Pudong New District from July 2006 to October 2010.The laboratory examinations were also performed including the urine routine,urinary protein to creatinine ratio,serum creatinine,blood glucose and lipids.The glomerular filtration rate (eGFR) was estimated by simplified Chinese MDRD (modification of Diet in Renal Disease) equation.SPSS 13.0 statistical software was used for statistical analysis.Results Among 1905 residents who completed survey and examinations,the adjusted prevalence of albuminuria was 12.0％ (95％ CI:0.105-0.135) and of eGFR less than 60 ml · min-1 · 1.73 m-2 was 1.9％ (95 ％ CI: 0.172-0.213).The prevalence of CKD was 12.6％ (95 ％ CI: 0.112-0.142).Logistic regression analysis revealed that age (OR =1.043),hypertension (OR =2.272),diabetes mellitus (OR =1.233)and hyperuricemia (OR =1.003)were independently associated with CKD.Conclusions The prevalence of CKD in adult residents (≥45 years) from Shanghai Pudong New District is high.It is necessary to carry out early screening and to intervene risk factors of CKD in middle-aged and older residents.
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Bilingual teaching is an important method to train international medical students.While the current bottleneck problem is the shortage of teachers during the exploration and practice in bilingual teaching. Proceeding from requirements and current situation of bilingual teachers, the following approaches should be taken to strengthen the teaching stuff construction of bilingual teaching: demonstration teaching, language training infiltrated in clinic job, guiding foreign students, preparing bilingual teaching materials and professional team building.
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Objective To assess the effect of continuous renal-replacement therapy (CRRT) dose on the outcome of acute renal failure (ARF) patients with meta-analysis of randomized controlled trials (RCTs). Methods Studies were identified by systematic search of peer-reviewed publications in Medline, EMBASE and Cochrane library database through June 2010. All the RCTs that compared the incidence of clinical outcome such as mortality, need for chronic dialysis between standard and low dose CRRT were eligible. The pooled relative risk (RR) for clinical outcome was compiled using a random-effects model. Heterogeneity was evaluated by means of subgroup and sensitivity analysis. Results Six eligible studies were identified. By meta-analysis, standard dose CRRT was associated with non-significant 13% mortality risk reduction (RR 0.87, 95%CI 0.70-1.07, P=0.19)and 13% composite outcome risk reduction of chronic dialysis dependence and mortality (RR 0.87, 95%CI 0.69-1.09, P=0.21), but the trend toward increased chronic dialysis dependence risk among survivors (RR 1.43, 95%CI 0.94-2.18, P=0.09). The overall test for heterogeneity among cohort studies was significant (P=0.001, I2=76.2%). The risk of mortality was modality was significantly lower in some studies of which delivered dose was moer than 35 ml·kg-1·min-1,modality was continuous venous-venous hemofiltration (CVVH) and major cause was non-sepsis treated with standard dose CRRT. Conclusions Standard dose CRRT in patients with ARF does not improve survival, renal recovery and composite outcome, but decreases mortality in important subgroups including those with higher delivered dose, CVVH and non-sepsis.
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Objective To investigate the risk factors for cardiovascular complications in the elderly persons undergone with dialysis.Methods One hundred and eighteen patients undergone with hemodialysis(HD)or continuous ambulatory peritoneal dialysis(CAPD)at the Eastern Hospital, Tongji University, Shanghai during January 2002 to December 2006 were retrospectively analyzed.They were divided into four groups based oil their age and mode of dialysis, i.e., 44 patients in the elderly HD group(aged over 60years), 16 patients in the elderly CAPD group(aged over 60), 46 patients in medium-age HD group(aged under 60 years)and 12 patients in the medium-age CAPD group(aged under 60).Routine biochemical parameters were measured for all the patients and their difference was compared among the four groups.Logistic regression analysis was performed for the risk factors of cardiovascular complications.Results Pulse pressure(PP)was(66±21)mm Hg, serum level of triglycerides(TG)was(1.28±0.67)mmol/L, serum level of lipoprotein(a)[LP(a)]was(434±202)mg/L, C-reactive protein was(CRP)(21±15)mg/L, serum phosphorous(P3-)was(2.02±0.77)mmol/L, and product of serum calcium and phosphorous(Ca×P)was(4.3±1.6)mmol2/L2, respectively, in the elderly group with dialysis, all significantly higher than those in the medium-age group with dialysis(t=1.535, 1.016, 2.154, 2.252, 4.676, and 4.534, respectively, P＜0.05).Incidence of cardiovascular events was markedly lower in the CAPD group than that in the HD group(P＜0.05), either in the elderly or medium-aged patients.And, incidence of cardiovascular complications correlated positively with age, PP, LP(a), CRP and Ca×P of the patients with dialysis, but correlated reversely with serum level of albumin(all with P＜0.05).Conclusions There exist a few risk factors for cardiovascular complications in the elderly patients with dialysis, so proper mode of dialysis should be selected for those who need it and adequate intervention should be implemented for them to correct the risk factors.
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Objective To investigate the oxidative DNA damage in lymphocyte of MHD patients and the effect of dialysis membrane on it through comet assay.Methods From 2003-05 to 2004-10 we selected 20 CRF patients undergoing maintenance hemodialysis and divided the patients into HE and PS group randomly.And 7 volunteers were chosen as controls.The patients in each group had undergone hemodialysis for 12 weeks using HE or PS membrane respectively.Then we took venous blood sample and separated lymphocyte.The oxidative DNA damage was measured by comet assay.Results The comet tail length of control group,HE group and PS group was(0.286?0.194)?m,(0.970?0.120)?m and(0.656?0.187)?m respectively.Compared with control group,the comet tail length of MHD patients was significantly longer(P
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Objectives To evaluate the function of peripheral blood T cell subsets in uremia patients and the influence of hemodialysis on them.Methods Expression of CD3,CD4 and CD8 antigen on peripheral blood T cells from 30 cases of hemodialysis and non-hemodialysis patients was evaluated by flow cytometry.Fifteen volunteers were also examined as normal controls.Results The expression of CD3,CD4,CD8 and CD4 +/CD8 + ratio in non-hemodialysis group(NHD) was significantly lower than that of control group(P
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Objective： Our aim was to purity the modifier protein of glyceraldehyde-3-phosphate dehydrogenase (G3PD) from African green monkey Vero-E6 line. Methods：Exposure of Vero-E6 cells to medium with a reduced K concentration (3.2 mmol/L) stimulated the growth and activation of G3PD. The increase of enzyme activity was mediated by a cytosolic modifier protein that was purified using affinity and anion-exchange high-performance liquid chromatograph. Results：The apparent molecular mass of the protein was 62 kDa. Western blotting and quantiative enzyme-linked immunosorbent assay showed that the amount of modifier protein increased progressively for 2 hours in cells exposed to low-K+ medium, and then returned to the control value, a kinetic profile similar to that the modifier protein is a constituent of renal epithelial cells and accummulated transiently in the low-K+ mitogenic signal. Conclusion: We obtained a modifer protein from monkey kidney epithelial cells (Vero-E6). It could activate G3PD and cell growth.
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Objective To investigate the inhibition effect of plasma-mediated small interfering RNA (siRNA) on the expression of monocyte chemotacite protein-1 (MCP-1) gene in human renal tubular epithelial cells (HKC).Methods Three pairs of siRNAs directed human's MCP-1 mRNA 67,116,142 targets were designed and synthesized.Eukaryotic expression vector special for MCP-1,pRNAT-MCP-1-Ⅰ、pRNAT-MCP-1-Ⅱ、pRNAT-MCP-1-Ⅲ were constructed and transfected into HKC by lipofectamine.At 24 hour after transfection,the expression of MCP-1 in the levels of mRNA was detected by Real Time RT-PCR,and the expression of MCP-1 in the levels of protein was detected by Western blot.Results Transfection efficiency of siRNA expression vector was 60%;the expression of MCP-1 in the levels of mRNA and protein of three pairs of plasma-mediated siRNA group were markedly decreased compared with normal control group(P
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Objective To elucidate the effects of Ginkgo biloba extract on advanced glycation end products-induced TGF-?_1,CTGF mRNA expression and oxidative stress in cultured NRK-49F cells.Methods From Jan.to Jul.2005,NRK-49F cells were incubated with serum-free medium with or without 25 mmol/L glucose as control.Cells were incubated with medium containing AGEs-BSA with or without 25 mmol/L glucose for 24 h.Cells were pretreated with EGb(50,100mg?mL -1)for 24 h to evaluate effects of EGb.And then intracellular ROS generation was measured by flowcytometry,expression of TGF-?_1 and CTGF mRNA were performed by a semi-quantitative RT-PCR technique.Results Compared with control,AGEs significantly increased the expression of TGF-?_1 and CTGF mRNA and intercellular ROS generation in NRK-49F cells.Incubation with both AGEs and high glucose(25 mmol/L)can upregulate TGF-?_1 and CTGF mRNA expression and intercellular ROS generation,compared with AGEs and high glucose(25 mmol/L)respectively.Preincubation with EGb attenuated AGEs-induced TGF-?_1 and CTGF mRNA overexpression and intercellular ROS generation.Conclusion Attenuation of AGEs-induced TGF-?_1 and CTGF mRNA overexpression in renal fibroblasts through scavenging of free radical and inhibition of oxidative stress may be one of mechanisms underlying EGb in prevention and therapy of DN.
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Objective: To determine the role of interleukin-10 (IL-10) secreted by peripheral blood mononuclear cells (PBMC) in the maintenance of hemodialysis in patients with immune deficiency and to evaluate thot if IL-10 can bea biocompatibility index of dialyzer. Methods: Twenty patients with end-stage renal disease were divided into maintenance hemodialysis group (MHD group) and non-hemodialysis group (NHD group). Patients in MHD group were dialyzed with cellulose (CA group) and polysulfone (PS group) dialyzers respectively. Ten healthy persons were as controls (CON group). The levels of IL-10 in supernatants of PBMC cultured with or without lipopolysaccharide (LPS) were measured by using ELISA method in all groups. Results: There was no spontanous IL-10 secretion in all groups. The level of IL-10 in all supernatants secreted by PBMC cultured with LPS increased significantly, and the increased levels of IL-10 in MHD and NHD patients were lower than that in controls. The PBMC stimulated by LPS secreted more IL-10 in patients dialyzed by PS with better biocompatibility than those of CA dialyzers with poor biocompatibility. Conclusion: Patients immune deficiency could be improved by using better biocompatible dialyzers more than poor ones, particularly in humoral immunity induced by urimia. The IL-10 could be a parameter in evaluating the biocompatibility of dialyzer.