Objective To investigate the impacts of valsartan on cell apoptosis induced by angiotensin Ⅱ in vascular smooth muscle cells,and discuss whether the mechanism is relevant to AMP-Activated Protein Kinases.Methods Vascular smooth muscle cells (A7r5) were designated to 5 groups:①control (DMSO) group,②Angiotensin Ⅱ (Ang]Ⅱ) 100 μmo]/L group,③Angiotensin lⅡ 100 μmol/ L + valsartan 10 μmol/L group,④Angiotensin Ⅱ 100 μmol/L + valsartan 10 μmol/L + compound C 1 μmol/L group,⑤ Angiotensin Ⅱ 100 μmol/L + 5-Aminoimidazole-4earboxamide-ribo-nucle-oside (AICAR) 100 μmol/L group,after 24h incubation,the intracellular activity of Caspase 3 was measured by spectrophotometry,the cell apoptosis were enumerated by low cytometry,the intracellular AMP-Activated Protein Kinases (AMPK) phosphorylation and total expression quantity were examined by western blot,the intracellular reactive oxygen species (ROS) was measured by fluorescent probe DCFH-DA,the intracellular activity of total superoxide dismutase (SOD) was measured by WST-1 method,the intracellular activity of Malondialdehyde (MDA) was measured by TBA method.Two groups were compared by using Student t test.Differences among multiple groups were evaluated by ANOVA.Results Compared with control group,the cell apoptosis of Angiotensin Ⅱ group was increased [(45.46 ± 15.40)％ vs.(1.88 ± 3.28)％,P =0.002],the synthesis of ROS was increased [(9.24 ±0.46) vs.(1.00 ±0.00),P＜0.01],theactivity of Caspase 3 was increased [(35.03 ± 3.54) vs.(13.33 ± 1.79),P ＜ 0.01],the activity of MDA was increased [(4.32 ±0.73) vs.(2.05 ±0.18),P＜0.01)],the phosphorylation of AMPK was decreased,the activity of SOD was decreased [(90.29 ± 14.73) vs.(136.02 ± 18.82),P =0.001];compared with Angiotensin Ⅱ group,the cell apoptosis of Angiotensin Ⅱ + valsartan group and Angiotensin Ⅱ + AICAR group were decreased [(24.91 ±8.46)％ vs.(45.46±15.40)％,P=0.031];[(27.90 ±4.39)％ vs.(45.46 ± 15.40)％,P =0.038],the synthesis of ROS was decreased [(2.37 ±0.05) vs.(9.24±0.46),P＜0.01];[(2.79±0.31) vs.(9.24±0.46),P＜0.01],the activity of Caspase3wasdecreased [(18.08±2.69) vs.(35.03±3.54),P＜0.01];[(27.83±3.56) vs.(35.03 ± 3.54),P =0.002],the activity of MDA were decreased [(3.25 ± 0.55) vs.(4.32 ± 0.73),P=0.017];[(3.46±0.60) vs.(4.32±0.73),P=0.047],the phosphorylationofAMPKwas increased,the activity of SOD was increased [(140.71 ±20.27) vs.(90.29 ± 14.73),P ＜0.01];[(116.73 ± 17.96) vs.(90.29 ± 14.73),P =0.029];compared with Angiotensin Ⅱ + valsarntan group,the cell apoptosis of Angiotensin Ⅱ + valsartan + compound C group was increased [(43.84 ± 12.00) ％ vs.(24.91 ± 8.46)％,P =0.043],the synthesis of ROS was increased [(4.64 ± 0.15) vs.(2.37 ± 0.05),P ＜ 0.01],the activity of Caspase 3 was increased [(25.64 ± 3.52) vs.(18.08 ± 2.69),P=0.011],the activity of MDA was increased [(5.12 ±0.92) vs.(3.25 ±0.55),P＜ 0.01],the phosphorylation of AMPK was decreased,the activity of SOD was decreased [(99.48 ± 16.59) vs.(90.29 ± 14.73),P =0.002)].Conclusions Valsartan could inhibit angiotensin Ⅱ-induced vascular smooth muscle cell apoptosis via activating AMPK,suppressing the synthesis of ROS and the activity of MDA,elevating the activity of SOD.

Root canal biofilm is frequently detected in the canal wall of infected root canal and the root canal with failed root canal therapy. Due to its special structure and diverse composition, root canal biofilm has the ability of the drug tolerance and antiimmunity, which lead to apical periodontitis. This review summarizes the features of the root canal biofilm and latest clinical methods to remove it.

Objective To summarize the clinical experience of laparoscopic repair of esophageal hiatal hernia.Methods A total of 15 cases of esophageal hiatal hernia underwent laparoscopic hernia repair and fundoplication from May 2004 to April 2005 in this division.There were 4 cases of type Ⅰ hernia(all of which presented severe gastroesophageal reflux disease),10 cases of type Ⅱ hernia,and 1 case of type Ⅲ.Surgical procedures included laparoscopic Nissen total fundoplication in 9 cases,Toupet partial fundoplicatin in 4 cases,and Dor partial fundoplication in 2.Symptoms of gastroesophageal reflux disease,including heart burn,dysphagia,regurgitation,chest pain,and belching,were evaluated by using the Visual Analogue Scales(VAS) at preoperative period,1 postoperative month,and 6 postoperative months,respectively.Results No conversion to open surgery was required in this study.The operative time ranged 100~187 min(mean,125 min).The postoperative hospital stay was 2~5 days(mean,2.8 days).All the patients were followed for 1~12 months(mean,8.5 months).No hernia recurrence was found.The VAS scores decreased from 5.0?3.9 preoperatively to 0.9?1.3 at 1 month postoperatively(t=3.823,P

Objective To observe the change of insulin-like growth factor 1 (IGF-1)before and after glucocorticoid (GC) therapy and to explore the effect of its change on bone metabolism in primary nephrotic syndrome (PNS) patients.Methods A total of 39 PNS patients with mean age of (36.73±12.15) years received GC therapy were selected from January 2008 to August 2009 in our hospital.Serum IGF-1,albumin,calcium,phosphorus,parathormone (PTH),25hydroxy vitamin D3,bone gla protein (BGP),degradation products of C-terminal telopeptides of type I collagen (CTx),24-hour urinary protein excretion and the ratio of urinary calcium to creatinine were measured at five time points-before GC therapy,4 weeks,8 weeks,12 weeks and 24 weeks after the use of GC.BMD was also detected at the same time points.Correlations among indexes were analyzed by Pearson.Results Thirty-six PNS patients fulfilled the follow-up and had complete clinical data,while other 3 patients lost.After GC treatment,serum calcium and 25hydroxy vitamin D3 were significantly increased in a time-dependent manner and were negatively correlated with 24-hour urinary protein excretion (r=-0.749,r=-0.831,P＜0.05,respectively).Serum BGP and IGF-1 were decreased after GC therapy in a time-dependent manner while CTx was significantly increased until week 12 after treatment (P＜0.05).Compared with pre-treatment,BMD of various parts had no significant difference at week 4; BMD of lumbar spine (L1-L4) was significantly decreased until week 8 (P＜0.05); BMD of femoral neck and femoral shaft was significantly decreased at week 24 (P＜0.05).IGF-1 was positively correlated with BGP and BMD (r=0.896,r=0.495,P＜0.05) and negatively correlated with serum CTx (r=-0.697,P＜0.05 ).Conclusions Serum IGF-1 level decreases in a time-dependent manner after GC treatment,which is correlated to BGP,CTx and BMD.Glucocorticoid treatment affects bone metabolism through IGF1 pathway possably in patients with PNS.IGF-1 may be used as a new bone biochemical marker of glucocoritcoid - induced osteoporosis.

Objective To observe the mitochondrial damage associated with protein-energy wasting of skeletal muscle in diabetic kidney disease (DKD) model of Goto-Kakizaki(GK) rats and evaluate the effects of low-protein diet supplemented with α-keto acids on muscle wasting.Methods Forty-five male 24-week-age GK rats were randomly divided into three groups,normal protein diet group (NPD),low-protein diet group (LPD) and LPD +or-keto group (Keto).Fifteen gender and age matched Wistar rats were served as control group (CTL).The living condition of GK rats was observed and the weight was measured once a week.Urine albumin,serum glucose,creatinine and urea nitrogen were measured at 24,32,40,48 week age.Soleus muscle was observed to calculate the muscle size and the percentage of Ⅰ and Ⅱ type muscle fiber with software after SDH and NADH staining at 48-week-age.Tissue ultrastructure was observed under the transmission electron microscopy.The activity of citrate synthase was detected by spectrophotometer.Expression of mitochondrial DNA was examined by Q-PCR.Results Compared with the CTL group,NPD,LPD and Keto groups had lower body weight,higher urine albumin,higher serum creatinine and urea nitrogen (P ＜ 0.05).The crosssectional area of muscle fibers was larger in CTL group.Compared with CTL group,the muscle fiber was partly broken,the mitochondrial morphology was obviously changed,the percentage of type Ⅱmuscle fiber was increased significantly (P ＜ 0.05),and the activity of citrate synthase and the number of mitochondrial DNA were decreased significantly in NPD,LPD and Keto groups (P ＜ 0.05).In Keto group,muscle wasting was improved compared with NPD and LPD group (P ＜ 0.05),the crosssectional area of soleus muscle increased and the percentage of type Ⅱ muscle fiber decreased,levels of urine albumin,semm creatinine and urea nitrogen decreased (P ＜ 0.05).Under transmission electron microscopy,the muscle fiber of keto group was intact and mitochondiral morphology was close to that of CTL group.The activity of citrate synthase and number of mitochondiral DNA were higher as compared to CTL group (P ＜ 0.05).There were no significant differences between NPD and LPD group.Conclusions In DKD condition,protein degradation in the skeletal muscle is accelerated,mitochondrion is swelling,the number of mitochondrial DNA is decreased and mitochondrial function is impaired.Low-protein diet supplemented with α-keto acids can improve mitochondrial damage and muscle wasting induced by DKD.

Objective To study the association of bone metabolism with the degree of proteinuria in patients of chronic kidney diseases (CKD). Methods A total of 71 CKD patients diagnosed as primary glomerulopathy were randomly selected from 2008.1-2009.5 in the First People's Hospital of Shanghai. They were classified into three groups according to proteinuria:group A of 25 patients, proteinuria ＜1.0 g/24 h; group B of 16 patients, proteinuria 1.0-＜3.5 g/24 h;group C of 30 patients, proteinuria ≥ 3.5 g/24 h. Fifty-eight healthy persons were selected from our medical examination center at the same time as control. Serum albumin, calcium, phosphorus,PTH, 25 hydroxy vitamin D3, bone gla protein (BGP), degradation products of C-terminal telopeptides of type I collagen (CTx), 24-h urinary protein excretion, and the ratio of urinary calcium to creatinine (UCa/Cr) were measured. Bone mineral density (BMD) was detected by dualenergy X-ray absorptiometry. Results Compared with control group, serum levels of calcium [(2.23±0.08), (2.13±0.09), (2.04±0.06)vs (2.37±0.12)mmol/L], 25-(OH)D3 [(50.19±6.58), (47.78±6.69), (42.42±10.85) vs (56.34±8.34) nmol/L] were significantly lower and UCa/Cr was significantly higher in A, B, C groups respectively (all P＜0.05). In group B and C, BGP was lower [(18.69±7.35), (16.13±5.76) vs (22.88±6.21) μg/L] and CTx was higher [(413.59±114.93),(516.21±314.25) vs (304.53±234.15) ng/L] (all P＜0.05). BMD was lower only in group C [(1.028±0.090) vs (1.090±0.062) g/cm2, P＜0.05]. Pearson analysis showed that 24-h urinary protein excretion was negatively correlated with serum calcium and 25 hydroxy vitamin D3, and positively correlated with UCa/Cr. UCa/Cr was positively correlated with serum CTx and negatively correlated with BGP. 25-(OH) D3 was positively correlated with BGP and negatively correlated with CTx. Conclusion Bone metabolism disorder exists in CKD patients, presenting the decrease of bone formation and the increase of bone resorption, which is associated with as the degree of proteinuria, especially in patients with nephrotic syndrome.

Objective To study the change of serum insulin-like growth factor 1(IGF-1)in primary nephrotic syndrome(PNS)patients and its relationship with bone metabolism, and to investigate the clinical significance of IGF-1 in the mechanism of bone metabolic disorders in PNS patients. Methods A total of 30 PNS patients with chronic kidney disease(CKD)stage 1 and 2 were randomly selected from 2008.1 to 2009.5 in our hospital. Serum IGF-1, albumin, calcium, phosphorus, PTH,25 hydroxy vitamin D3, bone gla protein(BGP), degradation products of C-terminal telopeptides of type I collagen(CTx), 24-hour urinary protein excretion, and ratio of urinary calcium to creatinine(UCa/Cr)were measured. Healthy control group of 61 persons were randomly selected from our medical examination center at the same time. Results Serum levels of calcium, 25 hydroxy vitamin D3 and BGP were significantly lower;CTx and UCa/Cr were significantly higher in PNS patients(P<0.05)as compared to healthy control group. BMD of PNS patients was lower but without significant difference compared with healthy control group[(1.078± 0.090)g/cm2 vs(1.090±0.062)g/cm2, P>0.05]. Serum level of IGF-1 was significantly lower in PNS patients and was positively correlated with BMD and BGP,and negatively correlated with 24-hour urinary protein excretion and CTx. Conclusions Bone metabolic disorder exists in PNS patients with the appearance of decreased bone formation and increased bone absorption.Serum level of IGF-1 has good correlations with bone biochemical markers.which may be used as a new bone biochemical marker of bone metabolism in kidney disease.

Objective To investigate the expression of Notch 1 receptor in renal tissues of patients with hepatitis B virus associated-glomerulonephritis (HBV-GN) and its role in the pathogenesis of HBV-GN.Methods A total of 48 patients with HBV-GN confirmed by renal biopsy during 2008-2010 were enrolled in the study.Distribution of Notch1 receptor in renal tissue of HBV-GN was detected by immunohistochemistry and the association between the distribution of Notch1 receptor and HBsAg was examined by double-label immunofluorescence assays.Correlations of Notch1 receptor expression with renal pathology and clinical parameters of HBV-GN were analyzed.Results Notch1 receptor distributed mainly in renal tubular epithelial cells and interstitial area as brownish red granules,and a few expression in glomerulus was also found.The positive score of Notch1 receptor expression in HBV-GN patients was significantly higher as compared to primary glomerulonephritis patients with serum HBsAg positive or negative and normal renal tissue controls.Notch1 receptor expression was more obvious in membrano-proliferative glomerulonephritis (MPGN) and mesangial proliferative nephritis (MsPGN) patients,but there was no significant difference among the different pathology groups.Distribution of Notch1 receptor was consistent with the distribution of HBsAg and its intensity was positively correlated with renal interstitial fibrosis (r=0.473,P=0.001),tubular atrophy (r=0.690,P=0.000),inflammatory cell infiltration (r=0.616,P=0.000).Negative correlation was found between renal function and the intensity of Notch1 receptor (r=-0.393,P=0.006).Conclusions Notch1 receptor expression increases in the renal tissues of HBV-GN patients and distributes mainly in renal tubular epithelial cells and interstitium,which is consistent with the distribution of HBsAg.Its intensity is closely correlated with renal interstitial lesions and renal function.Abnormal expression of Notchl receptor in renal tissue of HBV-GN may be involved in the progress of HBV-GN.

Objective To observe the muscle wasting in diabetic kidney disease (DKD) model of type 2 and non-obese diabetes mellitus in Goto-Kakizaki (GK) rats,and to evaluate the effect of lowprotein diet supplemented with α-keto acids on muscle wasting.Methods Forty-five male 24-weekage GK rats were randomly divided into three groups:normal protein diet group (22％ casein diet,NPD),low protein diet group (6％ casein diet,LPD) and LPD + α-keto group (5％ casein + 1％ α-keto,Keto).Fifteen gender-and age-matched Wistar rats were served as the control group (CTL).The living condition of GK rats was observed and body weight was measured once a week.Urine albumin,serum glucose,lipids,albumin,creatinine and urea nitrogen were measured at the age of 24,32,40,48 weeks.Soleus muscle at the age of 48-week was observed to calculate the muscle size with software.Expressions of atrogin-1,MuRF-1 and MyoD,myogenin were examined by Q-PCR and Western blotting.Results Compared with the CTL group,NPD,LPD,Keto groups had lower body weight [(317.90± 13.81),(330.38±11.96),(390.44±12.25) g vs (429.43± 16.85) g,all P ＜ 0.05],higher urine albumin [(14.36±5.52),(8.12±4.61),(5.58±3.50) mg/24 h vs (0.61±0.16) mg/24 h,all P ＜ 0.05],higher serum creatinine [(81.50±7.88),(66.32±8.36),(63.44±8.21) μmol/L vs (24.43±6.15) μmol/L,all P ＜0.05] and urea nitrogen [(7.53±1.05),(5.63±1.40),(5.54±0.97) mmol/L vs (2.98±0.62) mmol/L,all P ＜0.05].The cross-sectional area of soleus muscle fibers was larger in CTL group.Compared with CTL group,the expression levels of atrogin-1 and MuRF-1 increased significantly (all P ＜ 0.05),and of MyoD and myogenin decreased significantly in NPD,LPD,Keto groups (all P ＜ 0.05).In Keto group after 40 weeks,muscle wasting was improved compared with NPD and LPD group [body weight (381.62± 15.82) g vs (331.50±17.58),(326.60± 13.43) g,all P ＜ 0.05],cross-sectional area of soleus muscle increased,levels of urine albumin,serum creatinine and urea nitrogen decreased (all P ＜ 0.05),the protein expressions of atrogin-1 and MuRF-1 decreased,and myogenin and MyoD were higher as compared to CTL group (all P ＜ 0.05).There were no significant differences between NPD and LPD group.Conclusions In DKD condition,protein degradation in the skeletal muscle is accelerated,the genes which control muscle atrophy are activated,and proliferation and differentiation of the muscle satellite cells are impaired.Low-protein diet supplemented with α-keto acids can improve muscle wasting induced by DKD.

Objective To explore the expression and role of Toll receptor 4(TLR4)in human proximal tubular epithelial cell line HK-2,infected by HBV. Methods The serum of HBV DNA copies between 107-108/ml was collected. Before and after infected by HBV DNA positive serum. the HK-2 cells' morphology and the expression of α-smooth muscle actin(α-SMA)were observed by microscopy and immunofluorescence, and the effects of different concentrations of lipopolysaccharides(U)S.TLR4-stimulating factor)and CLI-095(TLR4 Inhibitor)on the proliferation rate of HK-2 cells were observed by MTT assays. After HBV serum and 10μg/ml LPS and 5μl/ml CLI-095 acted on HK-2 cells,TLR4 protein expression was measured by immunofluorescence and Western-blotting assay, and HBsAg and HBeAg in cell culture medium were detected by ELISA. and HBV DNA copies by fluorescence quantitative PCR. Results The longer HBV infected HK-2 cells, the more irregular of the cells' shape, the fewer number of the cells were left. But compared with HBV infected after 24 hours, α-SMA was more expressed after HBV infected 12 hours. After infected by HBV serum in 24 hours.HK-2 cells' proliferation rate was positively correlation in a dose range of LPS, but was negatively correlated with the CLI-095(P＜0.05＝.The levels of HBsAg and HBeAg in cell culture medium were largest when the LPS concentration was at 10μg/ml and CLI-095 at 5μg/ml.The expression of TLR4 significantly increased in HK-2 cells treated with LPS compared with those with CLI-095.but HBV DNA levels and HBsAg and HBeAg expression levels were lower. Conclusions HBV infection may promote cell transdifferentiation and cell injury. The stimulation of HK-2 infected with HBV by LPS may upregulate the expression of TLR4 and reduce the copies of HBV DNA.