Objective To establish multi-drug resistant bladder (MDR) tumor T24 cell lines and to assess their resistant characteristics.To observe effect of genistein on doxorubicin (ADM) resistant cell lines T24/ADM.Methods Bladder tumor T24 cell line was exposed to ADM in the culture medium for the establishment of drug resistant cell lines:concentrations of ADM was stepwise increased for long exposure.Morphologic studies were performed with optical microscopy.Drug sensitivities were determined by MTT.Results Six months were taken to establish drug resistant cell lines T24/ADM.No obvious morphologic changes were observed between resistant and parental cell. But drug resistances to ADM, 5-Fu,cyclophosphamide and cisplatin were increased,and resistance index were 15.79,4.68,5.53 and 3.81,respectively.Among all groups,there were significant differences.After genistein was used to T24/ADM cells,the IC50 value of genistein was 40 μg/ml.The proliferation cells were induced by genistein at the concentration of 20-100 μg/ml. Conclusion Genistein can inhibit human urinary bladder cancer T24/ADM cell proliferation at some concentration.

Objective To explore the relationship between CD146(MCAM),microvessel density(MVD)in renal cell carcinoma(RCC) and its clinic-pathology,and to explore their correlation with clinic-pathologic parameter of RCC. Methods Immunohistochemisty was employed to determine the expression of CD146 and MVD in 43 RCC tissues and 20 normal control renal tissues. Results The positive expression of CD146 in RCC(90.7 ％,39/43) was remarkably higher than that in normal renal tissue(30.0 ％,6/20)(x2=27.77,P＜0.05).The expression of CD146 was not correlated with the category of RCC (x2=1.37,all P ＞0.05),but had a significant correlation to(the tumor volume x2=7.57)clinical stage(r=0.62) and metastasis of RCC(x2=19.99,P＜0.05). The MVD of RCC [(78.00±23.10)/200HP]was significantly higher than that of normal renal tissue [(23.05±7.93)/200HP].The MVD of CD146 was not correlated with the tumor volume and category of RCC (t=1.33,t=1.46,au P＞ 0.05),but had a significant correlation to clinical stage and metastasis of RCC (t=2.37,t=2.10,P＜ 0.05). There was a positive correlation between expression of CD146 and MVD in RCC(r=0.74,P＜0.05). Conclusion The overexpression of CD146 in RCC has a significant relation with tumor angiogenesis.The expression of CD146 and angiogenesis might serve as an important indicator of the development, progress and metastasis of RCC.

Objective To establish a multidrug resistance cell line of human bladder tumor and study its characteristics and mechanism of muhidrug resistance. Methods Human bladder tumor cell line T24 was induced to muhidrug resistance cell line T24/ADM by intermittent administration of high dose ADM. The multidrug resistance to multianticancer agents of the ceils was evaluated by MTT assay; the expression of MDR gene was assessed by RT-PCR and P-gp expression was determined by flow cytometry. Results T24/ ADM resisted to many anti-tumor agents, and its IC50 of ADM was 16.3 times higher than that of T24. Significant overexpression of MDR gene and p-gp of the multidrug resistant cells were detected. Conclusion T24/ADM is a human muhidrug-resistant cell line, and it's drug resistance relates to the overexpression of MDR gene and p-gp.

Objective To investigate the expression of TLR4 mRNA and NF-κB p65 in colorectal carcinomas and adjacent normal colon tissue, and evaluate their roles in the pathogenesis and development in colorectal carcinoma. Methods Sixty-three colorectal carcinoma samples and respective adjacent normal colon tissue samples ( well differentiated : 23 cases; moderately differentiated: 17 cases; poorly differentiated:20 cases; other differentiated type: 3 cases; lymph node metastasis: 27 cases; no lymph node metastasis:36 cases; Dukes A: 18cases;Dukes B: 14 cases Dukes C: 22 cases; Dukes D: 9 cases) were collected. The expression of TLR4 mRNA in colorectal carcinomas and adjacent tissue were detected by RT-PCR. The expression of NF-κB p65 was detected by WB. Results The expression of TLR4 mRNA in colorectal carcinomas and adjacent tissue were 86.42 ± 15.16 and 32.74 ± 9.44. It was significantly higher in carcinoma tissue than that in adjacent tissue ( t = 22.354, P ＜ 0.01 ). The expression of TLR4 mRNA in well, moderately and poorly differentiated coiorectal carcinomas were 69.58 ± 11.27, 64.57 ± 13.91 and 97.12 ± 15.44 respectively. TLR4 mRNA in poorly differentiated colorectal carcinomas was significantly higher than that in well, moderately differentiated ones ( t = 11.304 and 12.223, P ＜ 0.01 ). There was no difference between lymph node metastatic carcinomas ( 89.91 ± 13.33 ) and carcinomas without metastasis (81.16±13.59,t =0.959,P＞0.05). The expression of TLR4 mRNA in the Dukes A stage tumors (59.05±11.66) was lower than that in Dukes B(90.34 ±0.08),C(91.41 ± 15.21), D(101.46 ±17.43), respectively ( t = 8.708,9.664,9.525, P ＜ 0.05 ). The expression of NF-κB p65 in colorectal carcinoma(0.63 ±0.11) was significant higher than that in adjacent tissue(0.34 ±0.08,t = 18.266,P ＜0.01 ). The expression of NF-κB p65 in well, moderately and poorly differentiated colorectal carcinomas were 0.46 ± 0.09, 0.72 ± 0.11 and 0.77 ± 0.14, respectively. The experssion of NF-κB p65 in well differentiated colorectal carcinomas was obviously lower than the woderately and poorly differentiated carcinomas (t = 11.223 and 10.875, P ＜0.01 ). There was significant difference between the expression of p65 in lymph node metastatic carcinomas(0.82 ± 0.17) and non-metastatic carcinomas(0.57 ± 0.12, t =18.269,P＜0.05). The expression of NF-κB p65 in Dukes A colorectal carcinomas (0.39 ± 0.06) was lower compared with the Dukes B(0.72 ±0.12), C(0.69 ±0.14) and D carcinomas(0.76 ±0.13,t =10.442, 9.889 and 9.721, P ＜ 0.01 ). Conclusions The enhanced expression of TLR4/NF-κB p65 are closely associate with clinical stage and pathologic grade. NF-κB p65 may be an molecular marker of lymph node metastatic. The increased expression of TLR4/NF-κB p65 promote the pathogenesis and development of colorectal carcinoma.

Objective To study the effects of sensitized dendritic cells in the treatment of bladder tumor and further discuss the mechanism of this immunotherapy. Methods 44 female F344 rats, which irrigated N-methyl-N-nitrosourea into bladders every other week for a total of five doses, were induced to bladder tumor. They were treated subcutaneously with either PBS, unsensitized DC, freeze thawing supernatant of tumor cells, or sensitized DC respectively every week for a total of four times. In the fifteenth week, their bladders were weighted and harvested for observation by naked eye and microscope, their blood was harvested for examination CTL by FCM. Results The weight of bladders in sensitized DC group was lower than those in PBS group, unsensitized DC group and freeze thawing supernatant of bladder tumor cells group (P<0.05). The stages of bladder tumor in sensitized DC group were statistically descended compared with those in PBS group (P <0.05). The CD+3 T cells in sensitized DC group was lower than those in PBS group, unsensitized DC group and freeze thawing supernatant of bladder tumor cells group (P <0.05). The CD+3 CD+8 CD+28 T cells in sensitized DC group was higher than those in PBS group, unsensitized DC group and freeze thawing supematant of bladder tumor cells group(P <0.001). Conclusion Sensitized DC injecting subcutaneously can reduce the stages of F344 rats' bladder tumor, Unsensitized DC injecting subcutaneously has not effect in the treatment of bladder tumor;, while the effect of freeze thawing supematant of tumor cells injecting subcutaneously is not well. The mechanism of sensitized DC in the treatment of blader tumor is that DC plays an immunol killing role by presenting antigen, stimulating CTL.

Objective To investigate the effect of all trans-retinoic acid (ATRA) combining with Interferon α-2a on bladder tumor and the possible mechanisms. Methods Fifty female Wistar rats with bladder tumor were established and separated into 4 groups randomly that are DMSO group (A), IFN-α-2a group(B), ATRA group(C) and ATRA+IFN-α-2a group(D). Then each group was treated by drugs indicated. Finally, the weight of bladder, the stage and grade of tumor, the apoptosis and proliferation index of tumor were determined to estimate the effect of ATRA+IFN-α-2a. Results The body weights in group D are the highest and the bladder weights in group D are the lowest. The stages and grades of tumor in group D were statistically decreased compared with those in the other 3 groups. Accordingly, the apoptosis of cancer cells in group D was enhanced, whereas the proliferation index in group D was decreased significandy. Conclusion The effect of ATRA combined with Interferonα-2a on the bladder tumor is better than that of monotherapy.

With the development of economy and society, the change of heath concept and increased demands for health care , the cultivation of postgraduates majoring in clinical medicine is facing a lot of problems which seriously affects the quality of culti-vation.The article analyzed the problems and reasons in the clinical medicine postgraduate cultivation.We stated the competency of clinical medicine postgraduates according to the transition of health demands, global medical development and practical requirements for clinical personnel in China.We elaborated the required post competence for clinical medicine postgraduates on the basis of its con-tent and characteristics.We proposed to strengthen the cultivation of clinical medicine postgraduates by the reform and support of the government, enhanced construction of teachers, improved course plan and cultivation process, perfected assessment system in order to make the students competent to their posts.

Objective To explore the radical surgery for tetralogy of Fallot combined with right pulmonary artery stenosis.Methods Thirteen cases with tetralogy of Fallot combined with right pulmonary artery stenosis accepted radical surgery,among them 7 cases who-had a long right pulmonary artery stenosis accepted aortic transaction for radical surgery of tetralogy of Fallot;5 cases who had the entrance stenosis of the right pulmonary artery accepted right pulmonary artery patch expansion for radical surgery of tetralogy of Fallot;One case who had the distortion and stenosis after pulmonary shunt accepted the radical surgery for tetralogy of Fallot after seperation and expansion.Results There were no operation-related deaths,post-operative low cardic output and perfusion lung occurred for all the cases.Ten cases had grade 1 cardiac function and 3 cases had grade 2,respectively.Conclusion Right pulmonary artery stenosis increased the risks during the radical surgery for tetralogy of Fallot,but it is still the primary operation choice.

Objective To identify the expression of sphingosine-1-phosphate receptor (S1PR) subtypes, C-myc and His tag proteins of human umbilical vein endothelial fusion cell line, EA.hy926 and human umbilical vein endothelial cells (HUVEC), CRL-1730 for studying the function of apolipoprotein M (ApoM)-S1P axis. Methods Two kinds of cells (EA. hy926 and CRL-1730) were cultured to reach the density of 60%-70% in vitro. Immunofluorescence technique was em?ployed to investigate the expressions of coagulation factorⅧ(FⅧ), ApoM, S1PR1-S1PR5, C-myc and His tag proteins. Re?sults (1) Two kinds of cells both expressed FⅧand ApoM. FⅧpresented scattered particle distribution in CRL-1730, while uniform distribution in EA.hy926. However, ApoM was strongly expressed and widely distributed in cytoplasm of two kinds of cells. (2) S1PR1-3 can be detected on their membrane other than S1PR4 and S1PR5. S1PR1 was highly expressed but S1PR2 and S1PR3 were in a low level expression. (3) Two kinds of cells both expressed C-myc and His tag proteins in cytoplasm. Conclusion Two kinds of cells have the properties of endothelial cells and can express FⅧ, ApoM, C-myc and His tag proteins. It is not suitable for choosing C-myc and/or His tag–conjugated recombinant ApoM to study the fuction of ApoM-S1P axis with these two kinds of cells.

Objective To establish the orthotopic bladder cancer model of multidrug resistance as the human' s, and detect its resistance condition. Methods Two groups of nude rats 4-6 weeks of age were inculated with 1×107 cell of T24 or T24-ADM, following with observation and putting down their meat, drink,mental condition, urine and abdominal mass growth. Animals were sacrificed after 4 weeks later, then their bladder were weighted and measured, histopathologic assessment was performed,mdr1 was detected by PCR,and cells from the bladder tumors were detected of multidrug resistence by MTT. Results Group of nude rats inculated with T24-ADM generated tumors about 80 % (8/10), the one inculated with T24 was 90 % (9/10)and about 2-3 days early. The blank group had no rats emerge tumors in bladder mucosa at all. Bladder weight and volume: (0.8±0.3) g, (1.0±0.5) g, (875±158) mm3, (903±192) mm3, difference between the two groups had no significant (t = 1.332 and t = 1.215, P＞0.05). Histopathologic detection: The two groups of bladder cancer tissue biopsies can be seen more chaotic arrangement of cell structure, cell body shape is irregular, to the depth of myometrial invasion in different without breaking the film. Between the two groups there were no significantly differences. PCR detection of mdr1 expression differences between the two groups was significant (t = 3.612, P ＜0.01). Cytological detection of drug-resistant cell volume is slightly larger, and no significant difference in morphology. MTT detection: cells from the inculated T24-ADM mice bladder tumor were more resistance to ADM than the ones from the inculated T24 mice bladder tumor (F = 412.107, P＜0.01), and for several other drugs were also resistant. Conclusion Cell transplantation was successfully used to establish bladder cancer model in situ of T24-ADM, and with multi-drug resistance characteristics. The model laid the foundation for further multi-drug resistance research of bladder cancer.