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 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 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 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 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.

AIM: To determine the influence of interleukin-1?(IL-1?) and ?(IL-1?) gene polymorphisms on rheumatoid arthritis(RA) disease severity and secretion of IL-1?. METHODS: The study included 136 RA patients and 102 healthy controls. PCR-RFLP was used to detect site mutation at IL-1 gene. Meanwhile the IL-1? was also measured in the supernatant of the cultured and stimulated peripheral blood mononuclear cells(PBMC). RESULTS: No difference in the allele frequencies or genotypes of the IL-1? gene polymorphisms was found between the controls and RA patients.IL-1? allele 2 was overrepresented in patients with erosive RA but not in nonerosive patients. The patients with IL-1? allele 2 had a higher swollen joint index, higher tender joint index and erythrocyte sedimentation rate than those without IL-1? allele 2.The IL-1? in supernatant of stimulated PBMC from patients with IL-1? allele 2 had a higher level than that from those without allele 2. CONCLUSION: IL-1 gene polymorphisms may influence the occurrence of RA. Detection of IL-1? allele 2 have a potential prognostic value in RA.

Objective To identify the surface marker of bone marrow-derived liver stem cells and to isolate the stem cells, to investigate the differentiation of the stem cells. Methods The quantitative variations of the cells with stem cell surface markers, including ? 2-microglobulin negative (? 2m -), Thy-1 +,CD34 +,Flt-3 +,IL-3R +,and c-kit + markers, were detected by using flow cytometry in the bone marrow of several rat models with liver injury. Each stem cell population was then isolated using a magnetic bead cell-sorting procedure. The isolated cells were cultured in a system containing cholestatic serum and hepatocyte growth factor (HGF). The morphology of the cells was observed, and the expressions of albumin, AFP, and CK8/18 were detected with immunohistochemistry technique. Results ? 2m - cells elevated significantly in each of the rat models.After being co-cultured with cholestatic serum and HGF, ? 2m - cells showed multilateral transformation and resembled hepatcytes morphologically. The differentiated cells expressed albumin, AFP, and CK8/18, all known to be the characteristic markers of hepatocyte. The other cell population showed little quantitative changes, and did not express the same proteins. Conclusions The quantitative variation of ? 2m - cells corresponds to the severity of liver injury. ? 2m - cells have the ability to trans-differentiated into hepatocytes in vitro. They might be the marker of liver stem cells.

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro . METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.