Phosphorylation of beta-catenin tyrosine residue 654 plays an important role in the epithelial to myofibroblast transition (EMT). Introducing mimic peptide of tyrosine residue 654 domain of beta-catenin into cells may influence phosphorylation of beta-catenin tyrosine residue 654. To deliver this mimic peptide into renal epithelial cells, we used penetratin as a vector, which is a novel cell permeable peptide, to deliver hydrophilic molecules into cells. A tyrosine 654 residue domain mimic peptide of beta-catenin (PM) with fused penetratin was constructed, purified and then detected for the penetration of the mimic peptide into human renal tubular epithelial cells (HK-2). The results showed that purified fusion mimic peptide could efficiently and rapidly translocate into human renal tubular epithelial cells. It is concluded that a cell-permeable peptides mimic of tyrosine residue 654 domain of beta-catenin was successfully obtained, which may provide a useful reagent for interfering the human renal tubular epithelial-mesenchymal transition.
Carrier Proteins/*metabolism ; Epithelial Cells/cytology ; Epithelial Cells/*metabolism ; Fibroblasts/cytology ; Fibroblasts/metabolism ; Kidney Tubules/*cytology ; Peptides/metabolism ; Permeability ; Phosphorylation ; Tyrosine/*metabolism ; beta Catenin/*metabolism

Objective: To discuss the feasibility of a new method to evaluate wearing simulators,and to investigate the feasibility of specimens' depth loss as the index of the materials' wearing resistance.Methods: Two commercial composite resins(SureFil and Spectrum TPH) were selected.The specimens were subjected to 100,200,300,400,500,800,1200,1 600 and 2 000 cycles wearing in the CW3-1 wearing system.Wearing was determined quantitatively by weighing the specimen and measuring the height of the specimen,volume loss(mm 3) and height loss(?m) were calculated.Previous clinical study results on SureFil and Spectrum TPH conducted at Hong Kong University were used to examine the relationship between clinical wearing and laboratory wearing.The difference of the wearing resistance between the two materials and the Pearson correlation of the height loss and the volume loss were analyzed by statistical software SPSS 15.0.Results: The wearing resistance of the two materials was significant difference(P

Objective To investigate the correlation of p16 gene and p21 gene expression abnormal- ity with the prognosis of bladder carcinoma.Methods Using the search terms“bladder neoplasm”,“prognosis”,“p16”or“p21”,the literature on the correlation of p16 gene and p21 gene expression abnor- mality with the prognosis of bladder carcinoma were searched from MEDLINE database,PubMed database, CBMdisc and China Academic Periodical database,and were evaluated by Meta-analysis with Dersimonian- Laird model.Results A total of 19 trials involving 1584 patients(positive rate of 40.4%)were identi- fied,including 12 trials of 975 patients(positive rate of 37.4%)on p16 gene expression abnormality and 7 trials of 609 patients(positive rate of 45.4%)on p21 gene expression abnormality.The combined relative hazard(RH)of p16 gene expression abnormality on the prognosis of bladder carcinoma,p21 gene expression abnormality on the prognosis of bladder carcinoma and both p16 gene and p21 gene expression abnormality on the prognosis of bladder carcinoma was 3.70(95% CI,3.42-3.99),3.01(95% CI,2.81-3.21)and 3.18(95% CI,3.01-3.35),respectively.Conclusions Both p16 gene and p21 gene expression abnor- malities are biomarkers for poor prognosis of bladder carcinoma.The detection of these biomarkers may be helpful in making the treatment strategy.

Objective To describe a new kind of Y-shaped metallic stent delivery system and evaluate its feasibility and preliminary effect for managing multiple airway stenoses involving the lower trachea and the tracheal carina.Methods The Y-shaped metallic stent delivery system consisted of three- tier structure.The inner-tier was composed of four parallel guiding tubes,which was used for two guidewires and two threads passing through,the middle-tier was delivery catheter,which contained the four guiding tubes,and the outer-tier was introducer sheath.Under the fluoroscopic guidance,15 patients with multiple stenoses involving the lower trachea and the tracheal carina were treated with the new covered self-expandable Y-shaped metallic stents.Results Stent placement in the tracheo-bronchial tree was technically successful in all patients with obliteration of the dyspnea immediately after stent placement,and SaO_2 was increased form preoperative 75%—89% to postoperative 96%—99%.During follow-up a period of 3—58 weeks (M 22 weeks),all stenosis were resolved without stent-related complications,and the general physical of all 15 patents was improved with no occurrence of obviously dyspnea and bleeding.Karnofsky performance status(KPS)was improved from preoperative 26%—45% to postoperative 72%—95%.Five patients died of the following causes unrelated to stent insertion:multiple organ failure(n=3),cachexia(n=1)and pulmonary infection caused by gastrobronchial fistula(n=1),and the remaining 10 patients were alive with no evidence.of dyspnea at the time of this report.Conclusion Deployment of the covered Y-shaped metallic stent with the use of Y metallic stent delivery system in the management of airway stenoses involving the lower trachea and the tracheal earina was a simple and safe procedure and with a good short-term clinical efficacy.

Objectives： The current study was designed to identify mimic polypeptide epitopes of lung cancer-related antigen WLA-Ag1 through screening random peptide library which was on display in phages, and provide useful information for further study of the characterization of WLA-Ag1 antigen and the potential chance for developing new diagnostic or therapeutic methods for lung cancer. Methods： Through affinity enrichment and immunoscreening of phage-displayed fifteen-peptide libraries with a monoclonal antibody named WLA2C4 against WLA-Ag1, the enriched phages was obtained and plated on agar plates. Some positive clones picked at random were confirmed by ELISA using WLA2C4 monoclonal antibody and DNA sequencing. The homologous comparison of amino acid sequences of positive clones was conducted through searching the protein sequence databank on Internet. The digestion analyses of amino acid sequences were also conducted by a software. Results： Twelve positive clones have been found with a consensual DNA sequence, that is, 5′ -GCT GGT TGG ATT ACT TTT CAT CGT CGT CAT CAT GAT CGT GTT CTT-3′ . Amino acid sequence was AGWITPHRRHHDRVL, with three trypsinase-digestible sites. This coincided with our previous finding that WLA-Ag1 can be digested by trypsinase. Conclusions： The amino acid sequence found in this study might be the epitope of WLA-Ag1. Screening of bioactive peptides from random peptide libraries using monoclonal antibodies as the ligands is an effective method to identify the epitopes of cancer-related antigens.

Objective To investigate the expression of Erbin in renal interstitial fibrosis (RIF) and the effect of over-expression of Erbin on transforming growth factor β1 (TGF-(β1)-induced epithelial-mesenchymal transition (EMT) in NRK52E cells. Methods In vivo, the model of renal fibrosis was induced by 5/6 subtotal nephrectomy in rat. Scr and BUN was detected and Masson staining was used to evaluate the level of renal tissue fibrosis. The location and expression of Erbin in renal tissue were detected by immunohistochemistry and Western blotting. In vitro, after NRK52E cells were treated by TGF-β1 (10 μg/L) for 72 h, immunofluorescence and Western blotting were used to obverse the expression and distribution of E-cadherin and α-SMA. The expression of Erbin mRNA and protein were detected by RT-PCR and Western blotting respectively. NRK52E cells were transiently transfected with Prk5-myc-Erbin plasmid via lipofectamine 2000, then the expressions of Erbin, E-cadherin and α-SMA were detected by Western blotting. Results (l)Compared to sham group with Scr (33.96±7.28) μmol/L and BUN (8.11±2.55) mmol/L, rats in 5/6 nephrectomy model with Scr (140.52±61.11) μmol/L and BUN (34.23±7.66) mmol/L revealed renal dysfunction. Masson staining indicated kidney interstitial fibrosis, and the expression of Erbin was significantly increased in renal tissue(2.9 folds), especially in tubular epithelia. (2)In vitro, the expressions of Erbin and α-SMA were markedly increased (2.3 folds and 2.1 folds, P<0.05, respectively) and the expression of E-cadherin was dramatically decreased in NRK52E cells stimulated by TGF-β1, which were consistent with immunofluorescence results. TGF-β1-induced E-cadherin suppression and a-SMA induction could be efficiently blocked by over-expression of Erbin (all P <0.05). Conclusions Erbin is up-regulated in renal interstitial fibrosis, and over-expression of Erbin can partly inhibit renal EMT induced by TGF-β1, which indicates Erbin playing an protective role in renal fibrosis.

Objective To investigate the expression and distribution of Cdc42-interacting protein 4 (CIP4) in renal fibrotic tissue,and the interaction between CIP4 and β-catenin in transforming growth factor β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) model of HK-2 cell line.Methods In vivo,the model of renal fibrosis was induced by 5/6 subtotal nephrectomy in rat.Masson staining was used to evaluate the level of renal tissue fibrosis.The expression and distribution of CIP4 was detected by immunohistochemistry.In vitro,the EMT model of HK-2 cell line was induced by TGF-β1 (10 μg/L) for 72 h.Western blotting was used to observe the expression of E-cadherin,β-catenin,CIP4 and α-SMA.Colocalization and interaction of CIP4 and β-catenin were detected by immunofluorescence and immunoprecipitation respectively.Results Compared to sham group,CIP4 expression was increased in group of 5/6 subtotal nephrectomy,CIP4 was mainly distributed in basolateral side of renal tubular epithelia.In vitro,expressions of α-SMA and CIP4 were increased in HK-2 cells stimulated by TGF-β1 for 72 h (2.5and 1.8 folds,respectively) (all P＜0.05),expression of E-cadherin was decreased(P＜0.05).Partial colocalization between CIP4 and β-catenin was detected by immunofluorescence.In control group,CIP4 and β-catenin partially colocalized at the cell membrane.Mter the stimulation of TGF-β1,translocation to nucleus of CIP4 and β-catenin were increased,and partially colocalized in nucleus.The interaction between CIP4 and β-catenin was observed by immunoprecipitation in both control and TGF-β1 stimulated groups.Conclusions Expression of CIP4 in renal fibrotic tissue is increased,which is mainly distributed in basolateral side of renal tubular epithelia.CIP4 and βcatenin partially colocalize and interact with each other.CIP4 may play a role in EMT process through the interaction with β-catenin.

This study aimed to explore Semaphrin4D (Sema4D) expression and clinical significance in non-small cell lung cancer (NSCLC), and to define the roles and mechanisms of Sema4D in regulating the malignant behaviors of A549 cells by small interfering RNA (siRNA). Firstly, immunohistochemistry revealed that Sema4D was more frequently expressed in NSCLC than in lung benign lesion (P<0.05) and its overexprssion was associated with low differentiation (P<0.05), poor pTNM staging (P<0.05) and occurrence of lymph node (LN) metastasis (P<0.05). Endogenous Sema4D expression was suppressed by Sema4D siRNA in A549 cells overexpressing Sema4D. Protein levels of Sema4D, total Akt and p-Akt were examined by Western blotting. Cell proliferation, migration and invasion abilities were measured by MTT assay and Transwell assay respectively. Results showed that Sema4D siRNA significantly suppressed phosphorylation of AKT in A549 cells, but it did not alter total AKT expression. In addition, efficient down-regulation of SemaD significantly inhibit cell proliferation (P<0.05), migration (P<0.05) and invasion (P<0.05) in A549 cells. These findings suggest that Sema4D might serve as a reliable tool for early prediction of NSCLC poor prognosis. Sema4D could play an important role in promoting tumor proliferation, migration and metastasis in the NSCLC, by influencing the Akt protein phosphorylation. Inhibition of Sema4D may be a useful approach for the treatment of NSCLC.

Objective To investigate the feasibility and primary therapeutic effect of inverted Y-shaped self-expandable metal stent for complex airway stenosis.Methods On the standpoint of the peculiar anatomic structure and the pathological changes of complex airway stenosis,we designed the inverted Y-shaped self- expandable metal stent.Under the fluoroscopic guidance,7 stents were implanted in 7 cases of airway complex stenosis.Results The inverted Y-shaped self-expandable metal stents were placed seccussfully,with instantaneous relief of dyspnea and improvement of living quality.Conclusion The placement of inverted Y- shaped self-expandable metal stent is feasible and safe for treating airway complex stenosis.(J Intervent Radiol, 2007,16:92-94)

Objective To detect the expression of Beclin1 and Bcl2 in invasive pituitary adenomas and to explore the relationship of Beclin1 and Bci2 in invasive pituitary adenomas and the relativity between the 2 genes.Methods 61 specimens were classified into invasive group (32 cases) and non-invasive group (29 cases) according to the comprehensive evaluation of invasive pituitary adenomas.lmmunofluorescence analysis and RT-PCR were adopted respectively to detect the protein and mRNA expressions of Beclinl and Bcl2.The difference and relativity of Beclin1 and Bcl2 expression in invasive group and non-invasive group were analyzed.Results 32 specimens of pituitary adenoma were invasive and 29 were non-invasive.Beclin1 protein and mRNA expressions were lower in the invasive group than in the non-invasive group (P ＜0.01 ).Bcl2 protein and mRNA expressions were higher in the invasive group than in the non-invasive group (P ＜0.01 ).Pearson related analysis showed that Beclin1 mRNA expression was negtively correlated with Bcl2 mRNA expression in the invasive group ( r =-0.42,P =0.028 ).Conclusions Beclinl expression is decreased in invasive pituitary adenomas.The invasiveness of pituitary adenoma is closely related to the high expression of Bcl2 protein and mRNA,and the low expression of Beclin1 protein and mRNA.The inhibition of the autophagy may lead to the enhancement of the invasiveness of pituitary adenomas and that inhibition may come from the interaction of Beclin1 and Bcl2.