Objective:To study the expression of a novel inhibitor gene of apoptosis,survivin,in bladder transitional cell cancer,and its relationship with the expression of p53,bcl-2 and ki67 genes.Methods:Using streptavidin-biotin peroxidase(SP) method,we examined the expression of survivin,p53,bcl-2 and ki67 proteins in 10 normal bladder tissues and 58 bladder transitional cell cancer tissues.Results:Survivin was expressed in 41 of 58(70.7%) cases of BTCC.In contrast,normal bladder epithelium did not express survivin.Overexpression of survivin was related to the tumor grade and clinical stage.The expression of survivin was strongly correlated with p53.bcl-2 and ki67 expression.Conclusion:Apoptosis inhibition by survivin, alone or in cooperation with p53 and bcl-2 may participate in the onset and progression of bladder transitional cell cancer.Survivin could be a new diagnostic/therapeutic target in bladder tranditional cell cancer.

Objective: To study the expression of a novel inhibitor gene of apoptosis,survivin,in bladder transitional cell cancer,and its relationship with the expression of ki67 genes and recurence of bladder cancer.Methods: Using streptavidin-biotin peroxidase(SP) method,we examined the expression of survivin and ki67 proteins in 10 normal bladder tissues and 58 bladder transitional cell cancer tissues and followed up all of the cases,the time of follow-up survey was 11 to 48 months,with the average of 29 months.Results: Survivin was expressed in 41 out of 58(70.7%) cases of BTCC.In contrast,normal bladder epithelium did not express survivin.Overexpression of survivin was related to the tumor grade and clinical stage.The expression of survivin were strongly corelated with ki67 expression.The recurrence time of grade Ⅰ-Ⅱ bladder cancer in which expression of survivin was positive was 11?9 months,while the recurrence time was 29?15 months for negative expression of survivin in the of bladder cancer.Conclusion: Apoptosis inhibition gene survivin may participate in the onset and progression of bladder transitional cell cancer.High expression of survivin is correlated with hyperplasia and recurrence of bladder tranditional cell cancer.

Objective:To improve the differential diagnosis between benign renal mass and renal cell carcinoma(RCC) so as to lower the misdiagnosis rate.Methods:A retrospective analysis was done on 21 cases of renal mass,aged ranged from 33 to 63 with a mean of 51.Among the 21 years,All of them were preoperatively diagnosed as RCC by imaging.Results:All the cases were surgically treated.Three cases underwent enucleation or simple nephrectomy because of the diagnosis as benign renal mass by intraoperative frozen section.Eight underwent radical nephrectomy.And the other 10 underwent simple nephrectomy.The pathological analysis after operation found all were renal benign changes.None developed recurrence during the follow-up of 1 to 5 years.Conclusions:Some renal benign space occupying masses could be misdiagnosed as renal cell carcinoma only by imaging examinations and surgical exploration and intraoperative frozen section by pathologic study were the keys to lower the misdiagnosis rate.

Objective:To evaluate the efficacy and toxicity of docetaxel and cisplatin for treatment of hormone independent prostate cancer. Methods:Thirty-two cases hormone independant prostate cancer patients were enrolled in this study. All patients had received castration and/or anti-androgens but relapse with elevated PSA. Multiple metastasis were diagnosed by imaging approach. Before chemotherapy,there were 26 cases suffered from pain by bone metastasis. Docetaxel 75 mg/m2 + NS 250 ml was administered intravenously and cisplatin 100 mg/m2 +NS 500 ml was administered intravenously on the first days. Results:Levels of PSA decreased to normal(

BACKGROUND: Increased expression of survivin in various tumor tissues can regulate cell proliferation, division, and plays an important role in protecting cells from apoptosis.OBJECTIVE: To construct the specific micro RNA (miRNA) expression vector that can block the C_(57)BL mice survivin gene by RNA interference (RNAi) technique.DESIGN, TIME AND SETTING: The single sample observation was performed at the experimental center of Department of Neurology, The First Afliliated Hospital of Chongqing Medical University from June to November 2008.MATERIALS: Ring-shaped pcDNA~(TM)6.2-GW/EmGFPmiR and BLOCK-iT~(TM) Pol II miR RNA interfered Expression Vector Kit with EmGFP was produced by Invitrogen Company. DH5a E. coli was preserved at the laboratory. Xho I and BamH I enzyme, spectinomycin were provided by Shanghai Sangon Biological Engineering Technology & Services Co., Ltd.METHODS: According to sequence of mRNA of C_(57)BL mice survivin provided by Genebank, four pairs of specific oligonucleotide sequences were designed and synthesized by using the software. The annealed oligonucleotide fragment was sub-cloned into pcDNA~(TM)6.2-GW/EmGFPmiR expression vector by gene clone technique and transformed into DH5a E. coli, subsequently, a single colony was incubated into liquid medium containing spectionmycin. Finally the plasmid was extracted.MAIN OUTCOME MEASURES: The recombinant vector was identified by sequencing and agarose gel electrophoresis.RESULTS: The sequencing revealed that insertion element was correctly cloned into the vector without nucleotide mutation, absence or insertion abnormality. The result of double enzyme digestion demonstrated that the fragment length was coincidence with expectation.CONCLUSION: The C_(57)BL mice survivin miRNA expression vector is successfully constructed.

Objective To study the expression of E2F-3 and pRb in primary prostate cancer(PCa) and its clinical significance.Methods The expression of E2F-3 protein and pRb was detected in 49 PCa,20 benign prostatic hyperplasia(BPH),10 normal prostate tissues(NP) by EliVision~(TM) plus immunohistochemical staining.Results The positive expression rate of E2F-3 in PCa,BPH,NP was 63.27%(31/49),20%((4/20)) and 10%(1/10),respectively.The expression level of E2F-3 in PCa was significantly higher than that in BPH(P

Objective To identify whether cisplatin can induce autophagy of bladder cancer T24 cells and the possible mechanism, and to observe the relationship between outophagy and apoptosis.Methods MTT assay was applied to investigate the effects of various concentration of cisplatin( 0 , 10 , 20 and 40 μg/mL) on T24 survival.TEM was performed to detect the autophagosome formation .Western blot assay was used to analyze the expression changes of LC3-Ⅱ, P62 and extracellular signal-regulated kinase ( ERK1/2 ) and p-ERK at the protein level.The effects of autophagy on the survival and apoptosis of bladder cancer cells were investiga-ted.Results DDP observably inhibited proliferation of bladder cancer cells in a dose-dependent manner ( P<0.05), the 50% inhibiting concentration(IC50) was (30.3 ±2.4)μg/mL;DDP induced autophagy of bladder cancer cells, observably increased autophagosome induced by DDP; up-regulated expression levels of LC3-Ⅱproteins ( P<0.05 ) , down-regulated expression of P62 proteins ( P<0.05 );DDP increased the protein level of p-ERK (P<0.05); The inhibitor of ERK pathway U0126 inhibited DDP-induced autophagy, as evidenced by decrease in the expression of LC3-Ⅱproteins ( P<0.05 ) .After inhibition of autophagy by WTM in DDP-treated cells, cell viability was obviously decreased and apoptosis was increased (P<0.05);DDP combined with WTM observably enhanced cleavage of poly ADP-ribose polymerase 1 ( PARP-1 ) and cleaved-caspase-3 which is apop-tosis related proteins ( P<0.05 ) .Conclusions Autophagy can protect T24 cells against ciplatin-induced apop-tosis, the possible mechanism of autophagy is the ERK signaling pathway is activated .

Objective To assess the clinical efficacy of minimally invasive internal fixation combined with percutaneous kyphoplasty (PKP) in treatment of thoracolumbar burst fractures in the elderly.Methods Twenty-one cases of neurologically intact thoracolumbar burst fractures treated by PKP between January 2007 and December 2008 were included in this study.There were 8 males and 13 females at age of 65-78 years (means,70.6 years).Mean period from injury to operation was (3.7 ± 1.1) days (range,3-7 days).The injured segments included Ti1 in two cases,T12 in six,L1 in eight and L2 in five.Kphosis Cobb' s angle,correction degree of kyphosis angle,correction loss of kyphosis,perioperative indicators,visual analogue scale (VAS) and Oswestry disability index (ODI) were evaluated after operation.Results All the cases were followed up for a period of 26-56 months (mean 34.2 months).Operation averaged 98.7 minutes (range,80-120 minutes) and showed mean blood loss of 32.8 ml (range,30-85 ml).Ambulation started at mean 18.2 hours after operation (range,8-19 hours).VAS averaged (0.9 ± 0.6) points at postoperative one week.Postoperative X-ray films revealed mean 10.4° correction of kyphotic Cobb angle,followed by mean 1.8 °of loss in the follow-up longer than 24 months.According to hyperextension and hyperflexion radiographies,there was no abnormal activity of fixed segments,implant loosening and breakage or adjacent vertebral fractures.Conclusion Minimally invasive internal fixation combined with PKP can relieve pain and restore function in a short time and is thought to be a preferable treatment option for thoracolumbar burst fractures in the elderly.

Objective To study a case series of thoracolumbar flexion-distraction fractures combined with compression of anterior and central columns to analyze the causes of its occult symptoms and explore the injury mechanism,imaging diagnosis and operative reduction of the fractures.Methods A retrospective study was carried out on data of 39 patients with thoracolumbar flexion-distraction fractures admitted into our hospital from May 2003 to December 2006.Of all,there were 17 patients with compression of anterior and central columns.The diagnostic value of imaging examinations such as X-ray,CT and MRI was evaluated.All patients were treated with posterior operation,in which simple distraction of the anterior and central columns was followed by compression and reduction of the posterior column from the rear axial direction.The clinical efficacy and safety of operation were assessed.Results The patients with compression of anterior and central columns accounted for 43.6% of all thoracolumbar flexion-distraction fractures.Before operation,six patients were misdiagnosed as simple compression fracture.Various kinds of imaging examinations detected the bone injury and/or ligaments complex injury of the posterior column at different degrees.The positive results on X-ray,CT,spiral CT multi-planar reconstruction (MPR)and MRI were in 8,7,11 and 17 patients respectively.After simple posterior distraction in 17 patients,there occurred over distraction in eight patients who were cured with posterior recompression plus reduction.Conclusions The posterior column injury of thoracolumbar flexion-distraction fractures combined with compression of anterior and central columns is relatively occult and easy to be misdiagnosed as simple compression fracture,when MPR CT and MRI are helpful for diagnosis.The operation with twostep reduction of axial distraction followed by compression can attain satisfactory and safe results.

AIM:To investigate whether oxidative stress is able to induce autophagy in mesenchymal stem cells (MSCs), and to explore the effects of autophagy on MSC proliferation and apoptosis under oxidative stress circumstance as well as the underlying mechanism for promoting the therapeutic effects of transplanted MSCs on treating diabetes mellitus e -rectile dysfunction ( DMED) .METHODS: Hydrogen peroxide ( H2 O2 ) was applied to simulate the oxidative stress cir-cumstance.The effects of H2 O2 at concentration of 0, 50, 100, 200, 400μmol/L on the viability of MSCs were tested by the method of Trypan blue exclusion and MTT assay respectively .The methods of MTT assay , Western blot and transmis-sion electron microscope ( TEM) were used to explore the effects of H 2 O2 on MSC apoptosis and autophagy .RESULTS:The proliferation of MSCs was obviously inhibited by H 2 O2 in a dose-dependent manner ( P<0.01) and the 50%inhibiting concentration (IC50) was (384.58 ±16.89) μmol/L.H2O2 induced apoptosis and autophay of MSCs .The proliferation rate of MSCs was suppressed by H 2 O2 significantly ( P<0.05 ) , with a further decline by blockade of autophagy ( P<0.05) whereas increased by blockade of apoptosis (P<0.05).H2O2 induced MSCs apoptosis obviously (P<0.05), with an augment of apoptosis ( P<0.05) by blockade of autophagy .Furthermore, the H2 O2 increased expression of cleaved caspase-3 and cleavage of poly ADP-ribose polymerase 1 (PARP1), Which were decreased by apoptosis blockade whereas were enhanced by blockade of autopahgy .CONCLUSION:Oxidative stress plays a dual role in MSC survival , which in-duces MSC apoptosis and autophagy .Moreover , blockade of autophagy intensifies MSC apoptosis .Therefore , it is a promis-ing method to ameliorate the effects of stem-cell based therapy on DMED by enhancing protective autophagy to increase the survival rate of transplanted MSCs against oxidative stress circumstance caused by diabetes mellitus .