Objective To analyze the methylation status of Vimentin gene in feces of colorectat cancer (CRC) patients,to explore the possibility and clinical significance of which as molecular marker for early diagnosis of colorectal cancer.Methods DNA was isolated in feces of 60 colorectal cancer patients hospitalized and treated in the First Affiliated Hospital of Zhengzhou University,17 colorectal adenoma patients underwent colon endoscopy in outpatient department and 30 normal volunteers.The methylation status of Vimentin gene was analyzed with methylation-specific PCR (MSP).Results In the feces of 60 colorectal cancer patients,the positive percentage of Vimentin gene methylation was 51.67％ (31/60).In the feces of 17 colorectal adenoma patients,the positive percentage of Vimentin gene methylation was 4/17.Vimentin gene methylation was detected in none of 30 normal controls.Conclusion The methylation of Vimentin gene in fecal is an early event in colorectal cancer progression.Analysis of Vimentin gene methylation in feces is expected to be one of the markers for early diagnosis of colorectal cancer.

Objective To explore the effect of neural cell adhesion molecule (NCAM) on adhesion,migration and morphology of mouse bone marrow-derived mesenchymal stem cells (BMSCs).Methods We isolated and cultured BMSCs from wild-type and NCAM gene knockout mice.The expression of NCAM was detected by Western blot and immunofluorescence.Wound healing and adhesion assays were used to detect cell migration and adhesion ability respectively.The morphological changes were observed and the expressings of protein β1 integrin,E-cadherin,β-catenin and N-cadherin were analysed by Western blot.Results The migration and adhesion of BMSCs were significantly reduced after NCAM gene knockout.Meanwhile,the expression of β1 integrin was lower than those in wild-type BMSCs (P<0.01).The morphology of NCAM gene knockout BMSCs changed from irregular to flattened,and expressed epithelial identification marker E-cadherin and β-catenin (P<0.05).However,the expression level of mesenchymal identification marker N-cadherin was decreased (P<0.01).Conclusions NCAM is involved in adhesion and migration of BMSCs via regulating the expression of β1 integrin.Furthermore,NCAMmay negatively regulate the mesenchymal-epithelial transitions of BMSCs.

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 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. [

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.

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 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.

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 discuss the effect of early enteral nutrition based on rapid rehabilitation nursing in perioperative nursing of patients underwent robot-assisted laparoscopic prostatectomy. Methods A total of 93 patients who would undergo robot-assisted laparoscopic prostatectomy from January to December 2016,were randomly divided into the control and experimental groups. Patients in the control group(n=52) received traditional nutrition support protocol,while patients in the experimental group(n=38)were given early enteral nutrition based on rapid rehabilitation nursing theory. Recovery of gastrointestinal function,status of nutrition support and economic index were compared between the two groups. Results The mean anal exhaust time and the mean defecation time in the experimental group were(2.06±0.41)d and(2.57±1.00)d respectively, which were significantly earlier than those in the control group(P<0.05). The mean time of parenteral nutrition and post-operative hospitalization days as well as the medical cost in the experimental group were significantly shorter than those in the control group(P<0.05). Conclusions Early enteral nutrition can effectively promote the recovery of gastrointestinal function of patients underwent robot-assisted laparoscopic prostatectomy,alleviate the post-operative discomfort,shorten the post-operative hospitalization days and reduce the medical cost.

Objective To explore the clinicopathological features and renal outcomes of primary IgA nephropathy (IgAN) patients with chronic tonsillitis.Methods Patients with biopsyproven primary IgAN admitted to The First Affiliated Hospital,Sun Yat-sen University from January 2006 to December 2011 were enrolled.The clinicopathological features and renal outcomes of patients with and without chronic tonsillitis were retrospectively compared.The primary outcome was progression to end stage renal diseases and/or doubling of serum creatinine.Results A total of 981 primary IgAN patients were enrolled and 98 patients (9.99％) had a history of chronic tonsillitis.Compared with patients without chronic tonsillitis,IgAN patients with chronic tonsillitis exhibited significantly higher prevalence of acute episodes of tonsillitis as a predisposition (P ＜ 0.001),higher serum IgA levels (P=0.012),and higher prevalence of macrohematuria (P=0.006).No significant difference in renal pathological features was observed in patients with and without chronic tonsillitis.Moreover,the renal outcomes were similar as regards IgAN patients with and without chronic tonsillitis.Conclusion IgAN patients with chronic tonsillitis had higher prevalence of acute episodes of tonsillitis and macrohematuria as well as higher serum IgA levels.However,IgAN patients with and without chronic tonsillitis showed no significant difference in renal pathological features and renal outcomes.