Objective: to explore the expression and clinical significance of delta N p63 protein in transitional cell carcinoma of the bladder(TCCB).Method: The immunohistochemical staining assay was conducted to detect expression of delta N p63 protein in 43 cases of TCCB and 11 cases of normal bladder epithelium.The relationship between delta N p63 protein expression and pathological types,clinical stages of TCCB was analyzed.Results:The expression rates of delta N p63 protein in TCCB and normal bladder epithelium were 100%(43/43)and 18.2%(2/11)respectively.The difference between the two groups was significant(P

ObjectiveTo explore the clinical-pathway-based teaching effect in teaching urological postgraduate students in department of urology.Methods36 students were divided to two groups:18 students are the clinical pathway based teaching group, and the other 18 students are traditional teaching group.Examination was used to compare the effect of each teaching method after 6 month.The examination including history collection,physical examination and case file writing,clinical case analysis,clinical operational skills test and basic theory.ResultsThe record of history collection,physical examination and case file writing and basic theory test in clinical pathway based teaching group is not different between the two groups,but the record of clinical case analysis and clinical operational skills test in clinical pathway group is better than the traditional teaching group.ConclusionClinical pathway based teaching can improve the teaching effect of clinical case analysis and clinical operational skills in urological postgraduate students,and can improve the strengthen of clinical thinking and working ability.

Objective To explore the influence of up-regulated Foxp3 on Treg function and kidney transplantation chronic rejection reaction in rat model.Method The kidney transplantation chronic rejection reaction rat model was established.The F344 kidney was transplanted to Lewis rats,and retroviruses highly expressing Foxp3 were constructed.The Banff 97 hierarchical diagnostic criteria were used to diagnose chronic renal allograft nephropathy (CAN).The rat models were divided into three groups by random number table.In experimental group,the pSCV-BsdRFP-FoxP3 retroviruses were injected into the rats via the tail vein after operation.In negative control group,the pSCV-BsdRFP retroviruses were injected into the rats via the tail vein after operation.In blank group,the normal saline was injected into the rats via the tail vein after operation.Enzyme-linked immunosorbent assay (ELISA) was used to detect the changes of interleukin-10 (IL-10) and tumor necrosis factor-β (TGF-β) immediate,1,2,3,and 4 weeks after operation.The rats were killed at 4th week after operation,and kidney tissues were taken out for pathological examination.Result The pathological changes of CAN were observed at 4th week.The typical chronic rejection change was seen at 12th week.The levels of IL-10 and TGF-β were increased,and reached the peak at 3rd week.The levels of IL-10 and TGF-β in experimental group were higher than in negative control group and blank group at 1st,2nd,3rd,and 4th week.At 4th week,obviously different degrees of intimal thickening,and mild hyperplasia of interstitial fibers,glomerular sclerosis and infiltration with lymphocytes and plasma cells were observed in the three groups.In the experimental group,the lesions were mildest,and apparent neointimal hyperplasia was found.Conclusion pSCV-BsdRFP FoxP3 retroviruses can reduce the kidney transplantation chronic rejection reaction in rat model,and have the potential treatment effect.

Objective To summarize clinical features and explore diagnosis and treatment of femoral intercondylar notch non-bony impingement syndrome. Methods 15 patients of femoral intercondylar notch non-bony impingement syndrome were identified during arthroscopic operation of 115 patients (120 knees) with restricted knee joint extention during Oct 2004 to Dec 2007. Among these 15 patients, there were 3 cases of Bucket Handle Tear(BHT), 1 case of ACL's cyst, 3 cases of ACL tibial avulsion injury, 3 cases of synovial incarceration, and 5 cases of synovial chondroma. 9 patients were diagnosed by MRJ and 2 by X-ray before operation. All 15 patients were confirmed under arthroscopy. Results 15 patients( 15 knees) were operated and followed up for a period of 4～24 months, mean 13 months. Mean Lysholm score was 65(range, 41～75) before operation and 89(range, 75～100) after operation. Joint extension restrict was 5～25 degrees (mean 8.1 degree) before operation and 0 degree after operation. Conclusion Arthroscopy could accurately diagnose femoral Intercondylar notch non-bony impingement syndrome. Pre-operation MRI was helpful for diagnosis. If MRI and X-ray showed negative findings, diagnostic arthroscopic examination could be applied. Good subjective and objective effects could be achieved with arthroscopic operation.

Objective To evaluate dynamic changes of chemokine CXCL9 and chemokine receptor CXCR3 to predict the early acute rejection in rats after renal transplantation. MethodsRenal allogeneic grafting was performed on Brown Norway or Lewis rats as donors and Lewis rats as recipients. CXCL9 in serum was measured by ELISA and CXCR3 mRNA in peripheral blood lymphocytes by RT-PCR at the time points of just before and 1, 3, 5, and 7 d after operation. Serum creatinine (Cr) was measured simultaneously. ResultsThe serum contents of CXCL9 and Cr, and CXCR3 mRNA level in allograft group were significantly higher than those in isograft group (P

Objective:To investigate the expression of miR-203a in bladder cancer (BC) cell lines (RT-112, T24, 5637, UM-UC-3) and evaluate the effects on BC cell proliferation and radiosensitivity.Methods:Mir-203a mimics, mir-203a inhibitor, CDK6 siRNA, CDK6 expression plasmid and corresponding negative controls were transfected into BC cells. Quantitative real-time PCR was used to detect the expression of miR-203a in BC cell lines and human bladder epithelial immortalized cell line (SV-HUC-1). CCK8 assay was used to investigate the regulation of miR-203a and cyclin-dependent kinases 6(CDK6) on the proliferation of BC cells. Colony formation assay was performed to assess the effect of miR-203a and CDK6 on the radiosensitivity of BC cells. The target gene of miR-203a was confirmed by luciferase reporter assay. The effect of miR-203a on CDK6 protein expression was detected by Western blot. Multi-group comparison was performed by one-way ANOVA and two-group comparison was conducted by t-test. Results:Compared with the SV-HUC-1 cells, the expression levels of miR-203a in RT-112, T24, 5637 and UM-UC-3 cells were significantly down-regulated (all P<0.05). Compared with NC group, overexpression of miR-203a significantly inhibited the proliferation of BC cells, whereas knockdown of miR-203a significantly promoted the proliferation of BC cells (both P<0.05). Compared with NC group, overexpression of miR-203a significantly increased the sensitivity of BC cells to radiotherapy, whereas knockdown of miR-203a significantly weakened the sensitivity of BC cells to radiotherapy (both P<0.05). CDK6 was the target of miR-203a. Compared with NC group, overexpression of miR-203a significantly down-regulated the expression level of CDK6 protein, whereas knockdown of miR-203a significantly up-regulated the expression level of CDK6 protein (both P<0.05). After overexpression of CDK6 in T24 and UM-UC-3 cells transfected with miR-203a mimics, the cell proliferation ability was significantly increased, whereas the sensitivity to radiotherapy was significantly decreased compared with mir-203a mimics (both P<0.05). After CDK6 was silenced in RT-112 and 5637 cells transfected with miR-203a inhibitor, the proliferation ability of cells was significantly decreased, whereas the sensitivity to radiotherapy was remarkably increased compared with miR-203a inhibitor group (both P<0.05). Conclusion:miR-203a can serves as a tumor suppressor gene to inhibit the proliferation of BC cells and enhance the radiosensitivity of BC cells.

Objective To isolate and purify Thy 1 low Lin Sca 1 + bone marrow stem subset cells by magnetic activated cell sorting(MACS). Methods Thy 1 low Lin Sca 1 + cells from mouse bone marrow were collected through three processes by MACS.After Lin + cells were removed,Thy 1 low Lin Sca 1 + bone marrow stem cell subsets were harvasted.The purity of the cells was analysed by FACS and the reclaimation rate was counted. Results The purity and reclaimation rate of Thy 1 low Lin Sca 1 + cells were 72.36% and 82.43% respectively, which equaled to the level of isolation and purification of CD34 + cells by MACS. Conclusions It is effective to isolate and purify Thy 1 low Lin Sca 1 + bone marrow stem cell subsets by MACS, and the purity and reclaimation rate of the cells are high.

Objective To research the relationship between free/total prostate specific antigen (f/t PSA), PSA density(PSAD) and prostate cancer(PCa), to explore healthy middle and old-aged,the patients with BPH, the scope of reference value of PSA,frce PSA(fPSA), f/t PSA and PSAD in patients serum with PCa. Methods To de-tect 307 cases of healthy gerontism male,236 of BHP and 41 of PSA and fPSA in patients with PCa,to calcuLate f/t PSA and PSAD, to investigate the dependability between PSA ,I"PSA ,f/t PSA ,P SAD and healthy male, the patients ofBHP,the PCa ,to determine the scope of reference value PSA,fPSA,f/t PSA and PSAD fitting for Chinese. Results PSA and fPSA in patients with PCa are obviously higher than normal control(P <0.01). f/t PSA is obviously lower than the BPH and normal control(P < 0.01). When PSA is 9.25 and f/t PSA is 20%, there are better value of clini-cal diagnosis,when PSAD is 0.18 (AUC = 0.635), we can get best aceuration of diagnosis. Conclusions f/t PSA and PSAD are better than PSA in clinical diagnosis. When PSA is more than 9.25,f/t PSA is less than 20% or PSAD is more than 0.18 is significance for diagnosis. The normal reference value of PSA less than 9.25,f/t PSA more than 20%, PSAD less than 0.18 is fit for Chinese.

Objective To establish a method for the determination of perchlorate in food by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).Methods The perchlorate residue in spices and condiments was extracted with water,that in vegetables and fruits was extracted with acetonitrile-water (1∶ 1,V/V),and that in meat,poultry,eggs,milk and aquatic products was extracted with acetonitrile-water (2∶ 1,V/V).The supernatant was cleaned up with C18 SPE (3 ml,200 mg),and the detection was carried out by UPLC-MS/MS with internal standardmethod for quantification.Results The calibration curve was linear in the concentration range of 0.3-20.0 pg/L (R2 ≥0.999),the recovery was in the range of 82.6％-108.6％,the relative standard deviation (RSD) was in the range of 1.0％-9.9％,and the limit of detection was 2.0 μg/kg for milk,and 10.0 μg/kg for other food.Conclusion The method was simple,accurate and highly sensitive,and suitable for the determination of perchlorate in food.

Objective To construct the KU80 inhibition cell model by RNAi in U2OS cell and to explore the relationship between the Ku80,telomeres and radiosensitivity in telomerase-negative tumor cells.Methods U2OS cells were transfected with the recombinant plasmids of pshRNA-K80 by the lipofectamine,and the stable transfected cell clones were selected by G418.After the selection,the cells were collected and analyzed by the flow cytometry.RT-PCR and Western blot were used to measure the expression of Ku80 and Real-time PCR was used to detect the length of telomeres.The radiosensitivity of U2OS was determined by clone formation array.Results The transfection efficiency of the positive cell clones detected by the flow cytometry was (83.23 ± 7.63) ％.The inhibition rate of the Ku80 gene transcription in the cell group with recombinant plasmid was(68.09 ± 1.16)％ and the inhibition rate of the Ku80 protein expression in the same group was (11.03 ± 2.45) ％.The results of Real-time PCR showed that the telomere length of the cell group with recombinant plasmid (1.07 ± 0.07) was significantly shorter than that of the control group (4.42 ± 1.30,F =38.58,P ＜ 0.05) and that of the empty plasmid group (4.11 ±0.84,F =38.58,P ＜ 0.05).Compared to the control group,the telomere length of the empty plasmid group did not changed(4.42 ±0.84 vs.4.11 ±0.84).U2OS cells with Ku80 expression suppressed had lower SF2 than that of the control cells (F =1089.61,P ＜0.05),and resulted in the SER of 1.47.Conclusions The Ku80 inhibition cell model in telomerase-negative U2OS cell line is successfully established which has the shorter telomere length,and is more sensitive to radiation.Telomere shortening caused by pshRNA-of Ku80 is likely to be one of the mechanisms of radiosensitization in this kind of cell model.