Background and purpose:PTEN mutation has been found in 20%-40% of malignant gliomas.The common mutant epidermal growth factor receptor(EGFR vⅢ)was reported to coexpress in PTEN-deficient EGFR-expressing tumor.PTEN has been shown to interact directly with FAK and reduce its tyrosine phosphorylation levels to inhibit cell invasion.The invasion of glioma cells with EGFRvⅢ expression and PTEN deficiency is increased.This study was to observe whether PTEN inhibits glioma cell invasion even in the presence of strong pro-invasive signals provided by constitutive EGFR activity.Methods:U87?EGFR cells were transfected with pcDNA3.1 constructs encoding PTEN and the cells invasion levels were detected by transwell invasion assay.The expression of FAK was detected by immunoblotting.FAK expression vector was transfected into U87?EGFR-wtPTEN cells and the change of cells invasion was documented.Results:PTEN and PTEN(G129E)could inhibit cell invasion induced by EGFRvⅢ.PTEN and PTEN(G129E)could decrease the FAK phosphorylation at Tyr397.Over expression of FAK in U87?EGFR-PTEN abrogated PTEN-induced down-regulation of the phosphorylation status of FAK and rescued cell invasion.Conclusions:PTEN could inhibit cell invasion induced by EGFRvⅢ by dephosphorylating FAK.

Objective To detect the serum prohibitin protein(PHB)level in children with renal interstitial damage and analyze the correlation between PHB and renal interstitial fibrosis(RIF). Methods Serum PHB protein levels were determined by Western blot analysis in 36 children with kidney diseases,and 30 healthy children were studied as control. Levels of BUN,Scr,and urinary microprotein series(including ALBU/Cr,NAGU/Cr,IgG U/Cr,α1-MU/Cr)were studied by automatic biochemical analyzer. Renal interstitial damage was semiquantitatively graded according to Katafuchi's method. The correlation between serum levels of serum PHB protein and those of BUN,Scr as well as urine microprotein were analyzed. Results Serum PHB protein was positive in children with diverse kidney diseases however it was negative in the normal controls(P ＜ 0.05). Serum PHB levels were significantly higher in children with proliferative glomerulonephritis than those with non-proliferative glomerulonephritis(P ＜ 0.05). Statistical analysis indicated that serum PHB levels positively correlated with the degree of tubulointerstitial lesions(r ＝ 0.868,P ＜ 0.001)as well as the glomerular injuries(r ＝ 0.753,P ＜ 0.001). And,serum PHB levels were also positively correlated with urinary microprotein including NAG(r ＝ 0.586,P ＜ 0.001)and IgG(r ＝ 0.341,P ＜ 0.001). Conclusions Serum PHB levels were significantly increased in children with kidney diseases and were positively correlated with the degrees of renal interstitial damage,suggesting that PHB might be a potential clinical marker for detecting tubulointerstitial lesions.

Objective To investigate the role of osteopontin (OPN) in the pathogenesis ot cholesterol gallstone formation in bile.Methods The nucleation time of OPN in model bile and human gallbladder bile was studied by the nucleation time assay,the effect of OPN on cholesterol crystal growth in model bile was examined by the cholesterol crystal growth assay.The effect of OPN on vesicle was detected by the transmission electron microscopy in model bile and gallbladder bile; then the content of OPN and calcium were detected via the commercial kits in human bile.Results Osteopontin prolonged nucleation time in a dose dependent manner in model bile and human bile,and this effect was correlated with calcium.Compared with control group,the nucleation times were prolonged by 1.50and 1.93 times in lithogenic bile at the concentration of osteopontin 50 μg/ml and 100 μg/ml (P＜0.01),respectively. Nucleation time were prolonged by 1.17 and 1.33 times in normal bile (P＜0.01) and by 1.29 and 1.48 times in model bile (P＜0.01),respectively.The rate of cholesterol crystals growth was not influenced by calcium ions,but inhibited by osteopontin in a dose dependent manner in the model bile.Furthermore,the formation,aggregation and fusion of vesicles were delayed by osteopontin in bile samples as indicated by the transmission electron microscopy.The concentration of osteopontin [(0.53± 0.08) mg/ml vs. (0.65 ± 0.14) mg/ml,P＜0.05] and the calcium ions [ (0.71 ± 0.17) mmol/L vs. ( 0.84 ± 0.08 ) mmol/L,P ＜ 0.05 ] were lower in lithogenic bile than in control.Conclusions Osteopontin can inhibit the cholesterol gallstone formation in model and human gallbladder bile as the anti nucleating factor.

Objective To investigate the role of osteopontin (OPN) in cholesterol gallstone formation.MethodsGallbladder bile was obtained from patients with cholelithiasis (n=36,the experimental group) and from donors of liver transplantation (n=19,the control group).OPN,calcium ion and lipid were analysed quantitively.The nucleating role of OPN in bile was evaluated using nucleating time (NT) approach.ResultsOPN inhibited cholesterol nucleation in a dose dependent manner.OPN (50 μg/ml and 100 μg/ml) prolonged NT by 48.90％ (91.51％) and 17.07％ (32.93％) in lithogenic and control bile,respectively.OPN (100 μg/ml) also inhibited the nucleating effect induced by calcium ion.Furthermore,a combination of OPN (50 μg/ml) and calcium prolonged NT by 75.78％ and 33.96％ in lithogenic and control bile,respectively.A combination of OPN (100 μg/ml) and calcium prolonged NT by 125.9％ and 62.26％ in the 2 groups.The contents of osteopontin and calcium were significantly lower in lithogenic bile than control bile (P＜0.05).On the other hand,the cholesterol saturation index and the contents of cholesterol,phospholipid and bile acid were significantly higher (P＜0.05).ConclusionsOPN inhibited cholesterol gallstone formation.It may be involved in the pathogenesis of cholelithiasis.

Objective To investigate the potential roles of dexamethasone (Dex) in podocyte motility,and to explore the mechanism of modulating α-actinin-4,nephrin.Methods Podocytes were divided into three groups:Dex group [1 μmol/L Dex +50 mg/L puromycin aminonucleoside (PAN)],PAN group (50 mg/L) and normal control group.Scrape wound assay and Transwell migration assay were used to detect cell motility.Filtering ratio of podocytes was measured by FITC labeled BSA.Real-time PCR and Western blotting were used to examine the expressions of c-actinin-4 and nephrin.Results From the scrape wound assays,the ability of wound repair in podocytes of PAN group was significantly increased (P＜0.01),and the number of migrating cells in this group also rose (P＜0.01).Compared to PAN group,podocytes in Dex group did not enhance the motility after treatment with the same dose PAN (P＜0.01).Real-time PCR and Western blotting showed that Dex could significantly inhibit the up-regulated expression of α-actinin-4 and nephrin induced by PAN.Conclusions Dex can relieve the enhanced motility induced by PAN.Its mechanism may be associated with the modulation of the expressions of α-actinin-4 and nephrin.

Objective To evaluate the diagnostic accuracy of ELISA kit for cholesterol gallstone. Methods The ELISA kit of 33?500 vesicular protein was established by sandwich method, and the concentrations of the protein in gallbladder bile were examined among cholesterol, pigmental gallstone patients and control groups. Results The gallbladder 33?500 vesicular protein (213?70) ?g/ml is much higher in cholesterol gallstone patients than in pigmental gallstone patients (72?55) ?g/ml and control groups (65?52) ?g/ml (F=60.9, P

Purpose To oberve the dynamics of liver granuloma and the relative changes of Thl/Th2cytokine levels in mice infected with Schistosoma japonicum, and investigate the role of Th1/Th2 in S.japontcum granuloma formation and regulation. Methods Liver granuloma measurement were performedby histological examination and the ELISA were used for the quantitative determination of IL-2, IFNr andIL-4 in murine serum and spleen lymphocyte culture medium at 0,4,6,8,10 and 12 wk after infection.Results At 6 wk liver granuloma formation appeared and at 8 wk liver granuloma peaked. After 12 wk livergranuhoma diminished obviously. Meanwhile, at 4 - 6 wk IL-2, IFNr and IL-4 began to rise, at 8 wk thelevels of Th1 cytokines IL-2 and IFNγ peaked and then declined, and at 8 wk the levels of Th2 cytokines IL-4 were rapidly enhanced and increased obviously with a prolongation of the infection duration.Conclusions The Th1cytokines IL-2 and IFNγ were correlated well with S. japonicum granulomaformation and vigour, and the Th2 cytokines IL-4 might play an important role in down-regulating egggranuloma reaction at chronic schistosomiasis by inhibiting the Th1cytokines.

ObjectiveTo provide clues to find a biomarker for early diagnosis, prognosis and therapy, as well as to understand the molecular mechanisms governing cancer progression. Methods Surgical specimens were obtained from 87 patients with histopathologically proven malignant or benign lesions. The differential protein profiles of these malignant and benign specimens were detected using two-dimensional electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Western blotting and immuno-histochemistry were used to validate the results. RT-PCR was used to detect the gene expression in the tissues. ResultsMrp14 was found to be overexpressed in the tumor tissues of gallbladder cancer and extra-hepatic bile duct cancer, and in the bile of patients with malignant biliary tract tumours. The result was further verified using Western blot and immuno-histochemistry. RT-PCR confirmed the overexpression of Mrpl4 at the gene level. Mrp14 is a potential biomarker for biliary tract neoplasms. ConclusionsThis is the first report which described the overexpression of Mrp14 in biliary tract neoplasms and further studies are needed to confirm our findings. Mrp14 may be a potential hiomarker for biliary tract neoplasms. It may provide important clues on the molecular mechanisms governing cancer progression.

OBJECTIVETo evaluate the pro-nucleating activity in 33.5 x 10(3) vesicular protein.

METHODSThe model biles were established according Kibe and Zhu. The pro-nucleating activity of 33.5 x 10(3) vesicular protein were examined by polarized light microscopy. The protein and its enzymatic deglycosylation and proteolysis fractions nucleation promoting activity were detected by cholesterol crystal growth assay.

RESULTS33.5 x 10(3) vesicular protein displayed apparent potency of pro-nucleation with activity of 0.310, and derived crystal growth curve indices It, Ig, Ic were presented as 0.57, 1.52, 1.63 respectively, but after treated by N-glycanase enzyme and pronase, no promoting activity were found.

CONCLUSIONThe 33.5 x 10(3) vesicular protein may be involved in the nucleation process of gallstone formation, which is regulated by its peptide and sugar chain.

Bile ; metabolism ; Cholelithiasis ; physiopathology ; Cholesterol ; metabolism ; Humans ; Macromolecular Substances ; Microscopy ; Models, Biological ; Protein Transport ; Proteins ; metabolism