AIM: To investigate the influence of dialysate on the cell morphology and the peritoneal membrane function in chronic peritoneal dialysis rats. METHODS: 40 SD rats were divided randomly into 4 groups. Except control group, other groups daily received infusion of 20 mL dialysate (4.25% dialysate(HG), 1.50% dialysate(LG), Riger's solution(RG)) respectively for 8 weeks. The peritoneal membrane function was investigated by peritoneal transport test, and the rat peritoneal mesothelial cells(PMCs)morphology was analyzed by the cell imprints. RESULTS: The intraperitoneal volume and net ultrafiltration in HG and LG groups were significantly lower, with D/P_(urea) significantly higher, and C_(urea) after 4 h of dialysis significantly lower than that in RG and control groups. The density of cell population from analysis of cell imprints in HG and LG groups was significantly lower, but the mean surface area were significantly larger than that in RG and control groups. These change had no difference between HG and LG group. Using the high glucose dialysate for 8 weeks significantly decreased the ultrafiltration volume ,which was significantly relate to the increasing of cell surface area (r=-0.896,P

AIM: To investigate the expression of Smad2 signal protein in peritoneal mesothelial cells and how transforming growth factor ?1 (TGF-?1) affects its expression. METHODS: Rat peritoneal mesothelial cells were cultured in different levels of TGF-?1 (0,1.25,2.5,10 ?g/L) for different time (0,5,15,30,60,120 min). Endogenous Smad2 expression was evaluated by RT-PCR and immunohistochemical assay. The alteration of subcellular location of Smad2 was determined by immunohistochemical assay. RESULTS: TGF-?1 induced Smad2 mRNA expression, which increased at 5 min, peaked at 30 min, and declined to baseline levels by 120 min, in a time-dependent manner. Smad2 mRNA expression induced by TGF-?1 was also in a dose -dependent manner. TGF-?1 induced Smad2 phosphorlylation and nuclear localization in both time-dependent and dose-dependent manner, which was concordant with mRNA expression. Smad2 translocated from cytoplasm to nuclear accumulation in response to TGF-?1, and peaked at 30 min. CONCLUSIONS: Smad2 is present in peritoneal mesothelial cells. TGF-?1 may activate Smad2 expression and translocation to nuclear in a time-dependent and dose-dependent manner. [

AIM: To investigate the role of Smad7 in the Smad2 expression induced by transforming growth factor-?_1(TGF-?_1) in rat peritoneal mesothelial cells(PMCs).METHODS: Rat PMCs were cultured at different doses of TGF-?_1 (0,1.25,2.5,10 ?g/L) for different time(0,5,15,30,60,120 min).PCDNA3-Smad7 was then transfected into cultured rat PMCs by lipofectamine,and the cells were stimulated like the above.Endogenous Smad2 and Smad7 expression was evaluated by RT-PCR and Western blotting.RESULTS: TGF-?_1 induced increase in Smad2 mRNA and protein expression at 5 min,peaked at 30 min,and declined to baseline levels at 120 min, which was in a time-dependent manner.TGF-?_1 also induced Smad7 mRNA expression at 5 min,and then declined,down to the lowest at 30 min,but at 60 min it increased again.Smad2,Smad7 mRNA and protein expression induced by TGF-?_1 were also dose-dependent.After transfection,overexpressions of Smad7 mRNA and protein in rat PMCs were observed,which did not decline with time.The expression of Smad2 mRNA significantly decreased by 33%,56%,67%,71%,63% and 57%(P

AIM:To determine whether human peritoneal mesothelial cells express CD40. METHODS:Human peritoneal mesothelial cells (HPMC) were harvested from spent peritoneal dialysis effluent and maintained under defined in vitro conditions. Expression of CD40, CD40L and intercellular adhesion molecule-1 (ICAM-1) on HPMC under normal culture or stimulation with interferon-γ (IFN-γ), tumor necrosis factor-α(TNF-α), interleukin(IL)-1 and LPS were detected by FACS analysis. The relationship between CD40 expression and ICAM-1 expression on HPMC was analyzed.RESULTS:HPMC cultured in vitro expressed CD40 constitutively. The surface expression of CD40 was markedly up-regulated following stimulation with IFN-γ, but not with TNF-α, IL-1 and LPS. CD40L expression on HPMC was not detected. The expression of ICAM-1 on HPMC was significantly increased by stimulation with IFN-γ, TNF-α, IL-1, LPS, respectively; the most effective inducer on ICAM-1 expression was IFN-γ. The CD40 expression co-localizes with the expression of ICAM-1 and there was positive correlation between CD40 and ICAM-1 expression on HPMC. CONCLUSION:HPMC functionally expressed CD40.

AIM:To investigate the regulatory role of nuclear factor κB (NF-κB) in the expression of interleukin-6 in mesangial cells (MC) induced by interleukin-1β.METHODS:Activation of NF-κB was measured by electrophoresis mobility shift assay (EMSA). RT/PCR and ELISA were used to detect IL-6 mRNA expression and IL-6 production, respectively.RESULTS:rhIL-1β could rapidly stimulate the activation of NF-κB in MC, and increase the expression of IL-6 mRNA and protein. PDTC, one of the inhibitor of NF-κB， could inhibit the expression of IL-6 in mRNA and protein in MC stimulated by rhIL-1β.CONCLUSION:IL-6 expression induced by IL-1β may be regulated by NF-κB in MC, NF-κB may modulate the immune-inflammatory reaction in glomerular disease.

AIM: To investigate the regulatory role of nuclear factor ?B (NF-?B) in the expression of interleukin-6 in mesangial cells (MC) induced by interleukin-1?.METHODS: Activation of NF-?B was measured by electrophoresis mobility shift assay (EMSA). RT/PCR and ELISA were used to detect IL-6 mRNA expression and IL-6 production, respectively.RESULTS: rhIL-1? could rapidly stimulate the activation of NF-?B in MC, and increase the expression of IL-6 mRNA and protein. PDTC, one of the inhibitor of NF-?B, could inhibit the expression of IL-6 in mRNA and protein in MC stimulated by rhIL-1?.CONCLUSION: IL-6 expression induced by IL-1? may be regulated by NF-?B in MC, NF-?B may modulate the immune-inflammatory reaction in glomerular disease.

AIM: To investigate the effect of 4.25%peritoneal dialysis solution (PDS) on CD40 expression in rat peritoneal mesothelial cells so as to reveal the potential mechanisms by which CD40-CD40 ligand (CD40L) interaction may be involved in the inflammation of peritoneal membrane. METHODS: Rat peritoneal mesothelial cells (MC) were harvested from the peritoneal cavity and maintained under defined in vitro conditions. Expression of CD40 on MC under normal culture or stimulation with 4.25%PDS or 4.25%PDS+IFN-? was detected by RT-PCR and FACS analyses. After activation of CD40 on MC with CD40 mAb, the expression of intercellular adhesion molecule-1 (ICAM-1) on MC was analyzed by FCAS. RESULTS: MC cultured in vitro expressed CD40 constitutively. 4.25%PDS markedly up-regulated the expression of CD40 mRNA and its protein. The expression of CD40 mRNA and its protein following stimulation with 4.25%PDS+IFN-? was significantly higher than 4.25%PDS alone. The expression of ICAM-1 on MC was significantly increased after activation of CD40 with CD40mAb.CONCLUSIONS: MC functionally express CD40. The up-regulated CD40 expression on MC following stimulation with 4.25%PDS may play an important role in local peritoneal defense mechanisms and may be involved in the chronic inflammatory process of the peritoneum.

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 investigate the clinical and pathological characteristics of IgA nephropathy (IgAN) with macrohematuria (MH).Method 1512 consecutive patients with biopsyproven IgAN diagnosed from January 2006 to December 2011 were enrolled,and divided into MH group and control group respectively,according to whether there existed episodes of MH before renal biopsy.The clinical and pathological characteristics were compared between two groups.Patients in MH group were then divided into three groups according to the interval from the last episode of MH to renal biopsy to clarify the concomitant clinicopathological changes associated with occurrence of MH.Results The rate of MH in history was 22.1％.MH group patients had significantly lower serum creatinine,slighter proteinuria,lower prevalence of hypertension and heavier microhematuria than control group (all P ＜ 0.001).The prebiopsy durations were similar in two groups (P=0.627).In MH group,chronic pathological indicators,including global/segmental sclerosis,tubule atrophy/interstitial fibrosis were all slighter (all P＜ 0.001),whereas activity indicators,including necrosis lesions,crescents and mesangial proliferation were all more severe compared with control group (all P ＜ 0.05).Those who underwent renal biopsy within 30 days of the last episode of MH had more severe proteinuria and microhematuria,higher prevalence of necrosis lesions,more severe crescents formation,and endothelial proliferation (all P ＜ 0.05).Conclusions IgAN patients with MH in history have relatively milder clinical and chronic pathological manifestations,however more active pathological changes especially in those who suffer episode of MH recently.

Objective To assess the incidence of extrauterine growth retardation (EUGR) in very low birth weight (VLBW) preterm infants and to evaluate the effects of nutritional support and morbidities on EUGR.Methods Data of VLBW preterm infants ＜ 34 weeks of gestation admitted to the Neonatal Intensive Care Unit (NICU) of the First Affiliated Hospital of Sun Yat-Sen University between January 1,2005 and December 31,2010 were reviewed.Those VLBW preterm infants were divided into the EUGR group (n=67) and the non-EUGR group (n=40).Perinatal data,growth data,nutritional information and morbidities were compared between the two groups.The incidence of EUGR in VLBW preterm infants was assessed and the associated risk factors were analyzed.Independent samples t,Chi-square and rank sum tests and Logistic regression analysis were used for statistical analyses.Results A total of 107 VLBW infants survived to discharge.The average gestational age in the EUGR group was much lower than that in the non-EUGR group [(30.0±2.1) weeks vs (30.9 ± 1.1) weeks,t=2.904,P=0.002].However,the incidences of small for gestational age (SGA) and maternal hypertension in the EUGR group was higher than that in the non-EUGR group [SGA:53.7％ (36/67) vs 15.0％ (6/40),x2=15.575,P ＜ 0.01; maternal hypertension:40.3％ (27/67) vs 20.0％ (8/40),x2=4.689,P=0.030].Standard deviation score (SDS) of birth weight and weight at discharge in the EUGR group was lower than that in the non-EUGR group [SDS of birth weight:(--1.9±0.8) vs (--1.1±0.7),t=5.418; weight at discharge:-2.6 (-3.0--2.0) vs-0.5 (-0.9--0.1),U=30.271; both P ＜ 0.01].The velocity of weight gain in the EUGR group was lower than that in the non-EUGR group [(12.0±4.4) g/(kg · d) vs (16.1±4.0) g/(kg · d),t=1.879,P=0.036],while the maximum percentage of weight loss and the age at maximum weight loss in the EUGR group was higher than that in the non-EUGR group [percentage of weight loss:(13.2± 1.7)％ vs (9.0± 1.6)％,t=12.832,P ＜ 0.01; age:(13.4±3.5) vs (10.9±4.3) d,t=3.113,P=0.001].The time to achieve full enteral feeds,3 g/(kg · d) protein and 120 kcal/(kg · d) calories intake were longer in than the EUGR group [(39.7 ± 8.2) vs (30.8±6.1) d,t=6.293,P=0.007; (21.4±5.8) vs (17.5±1.3) d,t=4.286,P=0.002; (28.4±6.0) vs (20.3±5.4) d,t=7.198,P=0.015; 1 kcal=4.184 k J].The cumulative caloric deficit and cumulative protein deficit in the first two weeks of life in the EUGR group were significantly higher than those in the non-EUGR group [(600.9±49.3) vs (536.4 ± 55.2) kcal/kg,t=6.082,P ＜ 0.01; (17.4 ± 0.8) vs (12.4 ± 0.8) g/kg,t=31.279,P=0.003,respectively].The incidences of late-onset infection and bronchopulmonary dysplasia (BPD) in the EUGR group was significantly higher than that in the non-EUGR group [77.6％ (52/67) vs 40.0％ (16/40),x2=15.300,P ＜ 0.01;38.8％ (26/67) vs 17.5％ (7/40),x2=5.330,P=0.040,respectively].The length of oxygen therapy and mechanical ventilation in the EUGR group were significantly longer than that in the non-EUGR group [(44.5 ±4.5) vs (32.5± 1.5) d,t=20.042,P=0.030; 9.5(6.5-44.0) d vs 6.2(5.0-35.5) d,U=19.195,P=0.004,respectively].Logistic regression analysis showed that SGA,gestational age,BPD,late-onset infection,time to achieve full enteral feeds and 3 g/(kg · d) protein intake and the cumulative caloric deficit in the first two weeks after birth were the independent risk factors for EUGR (all P ＜ 0.05).Conclusions EUGR remains a serious issue in VLBW preterm infants,especially SGA,in the NICU.An early aggressive nutritional strategy,prevention of BPD and infection,and improvement of perinatal care may facilitate a reduction in the occurrence of EUGR.