Objective To explore the inhibition effect of recombinant mutant type(mt) p27 gene adenovirus (Ad-mt p27) transfection on the proliferation of colorectal cancer cell line SW480. Methods The recombinant mt p27 adenovirus vector was transfected into SW480 cells, western blotting analysis was applied to determine the p27 expression, and cell cycle distribution was analyzed using flow cytometry, and growth curve was drawn to observe the cell growth. Results There was high expression of p27 protein in the transfected SW480 cells. 77 9% cells were blocked in G 1/G 0 phase in the transfected group, which the percentage of the G 1/G 0 phase of Ad-LacZ transfected group and normal control group were 27 5% and 25 29% respectively. Ad-mt p27 transfection markedly inhibited SW480 cell growth in 24h and the inhibition effects was maintained for 7 days. Conclusion Recombinant adenovirus could efficaciously trasfect mt p27 gene into SW480 cells, and obtain high level mt p27 exprerssion. mt p27 markedly bolcked SW480 cell cycle progression and prominently inhibited SW480 cell growth. The mutant p27 gene transfection may be a new way of gene therapy for colorectal cancer.

Background:In recent years,the incidence and mortality of colorectal carcinoma have been increasing rapidly. Diagnosing and treating cancer at gene level have become a new effective approach. Aims:To investigate the expression of S-phase kinase-associated protein 2(Skp2)and its relationship with vascular endothelial growth factor(VEGF)and tumor microvessel density(MVD)in colorectal carcinoma. Methods:Thirty-five colorectal carcinoma patients from March 2014 to March 2015 at People’s Hospital of Hubei University of Medicine were enrolled. Fifteen para-cancerous tissue samples were served as controls. The expressions of Skp2,VEGF and MVD value were determined by immunohistochemical SP, and their relationships with clinicopathological characteristics were analyzed. Results:The expression of Skp2 in colorectal carcinoma was significantly higher than that in para-cancerous tissue( P = 0. 000),and was correlated with tumor differentiation and lymph node metastasis(P < 0. 05),but not with gender,age and TNM stage(P > 0. 05). Expression of VEGF and MVD value were significantly higher than those in para-cancerous tissue(P = 0. 019,P = 0. 002),and were correlated with lymph node metastasis and TNM stage(P < 0. 05). Expression of Skp2 was positively correlated with the expression of VEGF and MVD value in colorectal carcinoma( r = 0. 569,P = 0. 000;r = 0. 481,P = 0. 017 ). Conclusions:The abnormal high expression of Skp2 is involved in the angiogenesis of colorectal carcinoma. Skp2,VEGF expressions and MVD value may be served as important markers for malignancy degree of colorectal carcinoma.

Objective To evaluate the long term outcome of the continent urinary reservoir constructed with ileocecum appendix. Methods From 1985 to 1992,continent urinary reservoir was constructed by using ileocecum appendix after radical cystectomy for 56 patients,of which 11 have been followed up for over ten years. Results The daytime continence rate was 90% and the night continence rate 80%.The average capacity of the urinary pouch has been 388 ml,and the mean pressure 28 cm H 2O.The average urine flow rate is 9.6 ml/s.No evidence of ureteral reflux and no hyperchloremic acidosis have been observed.Stone formation in the pouch occurred in 1 patient,pseudo valve in the posterior urethra in 2 and stricture of the posterior urethra in 1. Conclusions Continent ileocecum appendix pouch might be considered as an ideal form of urinary diversion,characterized by low pressure,large capacity,continent and in situ urination.

Objective To study the clinical effects of intra internal iliac arterial chemotherapy for invasive bladder cancer after operation. Methods From February 1997 to June 2000,32 cases of invasive bladder cancer underwent partial cystectomy and arterial infusion pump implanted into the intra internal iliac artery, for postoperative chemotherapy. Results The patients have been followed up for 5～42 months, with a mean of 26 months,27 of the 32 patients have been free from the tumor,metastasis to lung and liver was noted in 2.Recurrence occurred in 3?10?16?30 months after operation respectively.Few side effects were noted. Conclusions Intra arterial chemotherapy might serve as an adjuvant therapy for invasive bladder cancer after operation.

Objective To study the potential therapeutic effect of potassium antimonyl tartrate (PAT) on human colon cancer in transplanted tumor nude mice models. MethodsSixty transplanted animal models were constructed with colon cancer cell line SW480 injected in nude mice. Nude mice were then random divided into 4 groups ( n = 15) :Normal saline group, 5-Fu group and different dose of PAT groups [ (20 mg/( Kg · d) ,40 mg/( Kg · d) ]. The volume of mass was measured every 3 days. After final-administration for 24 hours, immunohistochemical staining was used to detect the expression of PCNA in colon cancer cells. ResultsAfter the use of PAT, the growth of mass slowed down. PCNA levels [ (63. 63 ±8. 88)％ ,(59. 13 ±6. 15)％ ,(33. 38 ± 12. 76)％ ] in SW480 cells was reduced by PAT( P <0. 05, P <0. 01 ). ConclusionPAT potentially inhibited the growth of colon cell lines and induced apoptosis of SW480 colon cancer cells.

Objective To explore the therapeutic efficacy of modified Taohong Siwu Decoctio n(TSD) combined with limited internal fixation and external fixation for the treatment of tibial plateau fracture of Schatzker type Ⅴand Ⅵ. M ethods A total of 31 cases of tibial plateau fractures of Schatzker type Ⅴ and Ⅵ treated with TSD orally combined with limited internal fixation and external fixation were enrolled into the study. Follow-up was carried out for the evaluation of therapeutic effect and postoperative complications. Results All of the cases received follow-up for 6 months to 3 years. According to Merchant scoring criteria, the therapeutic effect was excellent for 15 cases, good for 12 cases, acceptable for 3 cases and inferior for one case, with the excellent and good rate being 87.1%. One case had pin tract infection, and then was cured after symptomatic treatment. Conclusion For tibial plateau fracture of Schatzker typeⅤandⅥwith unsatisfactory skin state, we can perform limited internal fixation and external fixation to take the place of strong steel plate fixation with extensive exposure for surgical treatment, and then give postoperative oral use of TSD, which will be a safe and effective therapy for the fracture by promoting the relief of the clinical symptoms and signs of early fracture.

AIM: To study the inducing effect of human mutant p27 gene on apoptosis of the colorectal cancer cells. METHODS: The recombinant adenovirus Ad-p27mt was constructed to infect the colorectal cancer cell SW480. The inducing effect of Ad-p27mt on apoptosis in colorectal cancer cells was measured by flow cytometry, DNA fragment analysis and TUNEL method. RESULTS: Ad-p27mt was successfully constructed. When the multiplicity of infection (MOI) was ≥50, the infection efficiency reached 100%. After 24 h of infection, there was an apoptotic hypodiploid peak observed by flow cytometry before G_1 and there were apoptotic characteristic bands in the DNA electrophoresis. The apoptotic index detected by TUNEL method was 82.6?3.2 (Ad-p27mt group) and 5.0?3.5 (control group), respectively, the difference of which was significant (P

Objective To investigate the effect of cyclic tensile stress (CTS) on the metabolism inand apoptosis of rat chondrocytes.Methods Primary rat chondrocytes were cultured on a Bioflex plate for one day and then stretched cyclically for 24 hours at a frequency of 0.5 Hz using a Flexcell-5000T apparatus.The cells were divided into 5 groups according to their stretching ratio:0％ (the control group),2％,6％,10％ and 14％.After the stretching,Col Ⅱ,Aggrecan,MMP-13 and ADAMTS-5 mRNA were measured using qPCRs,and the NO and PGE2 levels were measured using ELISA kits.Moreover,TUNEL staining and Annexin V-FITC/PI were used to analyze the apoptosis of chondrocytes.Results Compared with the control group,the average levels of Col Ⅱ and Aggrecan mRNAdecreasedin 10％ and 14％ groups [(0.738±0.11) and (0.58±0.13),(0.75±0.11) and (0.55±0.09)].In those groups,the MMP-13 [(2.47±0.47) and (2.88±0.36)] and ADAMTS-5 mRNA level [(2.39±0.33) and (2.75±0.49)],the NO [(6.96±0.96) and (8.28±0.82)] and PGE2 level [(6.83±0.66) and (7.15±0.71)] had increased significantly.In the 6％ group the average levels of Col Ⅱ(1.76±0.30) and Aggrecan mRNA (1.93±0.14)of 6％ group were significantly higher than the control group,but the NO level of the former (3.07±0.20) was significantly lower than the control group (3.89 ± 0.33).The apoptosis rate of chondrocytesin 2％ and 6％ groups were (0.065±0.013) and (0.063 ± 0.147),without significant differences to that of the control group (0.045 ± 0.008).However,compared with the control group,apoptosis in the 10％ and 14％ groups [(0.135 ±0.026) and (0.184±0.020)] increased significantly.Conclusion The effect of cyclic tensile stress on chondrocyte metabolism and apoptosis was magnitude-dependent.Ten percent and 14％ CTS can increase the catabolism and apoptosis of chondrocytes.Ten percent and 14％ strain can increase the catabolism and apoptosis of chondrocytes.Cyclic 6％ strain can increase the anabolism of chondrocytes,but 2％ strain has no impact on metabolism or apoptosis.

BACKGROUND: The incidence rate of colon cancer is on an obvious increase. Previous treatments are primarily surgical therapy; radiotherapy and chemotherapy. Gene therapy has been applied in the clinical practice for treating colon cancer.OBJECTIVE: To construct replication deficient hp27mt recombinant adenovirus and detect its expression in SW480 cell in order to investigate the viability of gene therapy based on adenovirus and study the anti-tumor characteristic of mutant p27.DESIGN: A non-randomized controlled trial.SETTING: Department of Gastroenterology, Affiliated People' s Hospital of Yunyang Medical College, Institute of Clinical Medical Science of Yunyang Medical College; Department of Gastroenterology, Zhongnan Hospital of Wuhan University.MATERIALS: The experiment was conducted in the Institute of Clinical Medicine, Yunyang Medical College from January 2002 to September 2003. Restriction endo-nucleases Age Ⅰ, Nhe Ⅰ, Kpn Ⅰ, Pac Ⅰ and Pme Ⅰ;Taq polymerase; T4 DNA ligase; Western blot kit; mouse anti-human p27kipl polyclonal antibody, horseradish-peroxidase conjugated goat anti mouse second IgG monoclonal antibody; pORF9-hp27mt plasmid, adenovirus framework plasmid pAdeasy-1, shuttle plasmid pShuttle-CMV, LacZ recombinant adenovirus Ad-LacZ, liposome, colon cancer cell line SW480.METHODS: hp27mt was excised from vector pORF9-hp27mt by restriction endonucleases, and inserted into shuttle plasmid pShuttle-CMV after two cycles of subclone, forming plasmid pShuttle-CMV-hp27mt. Then it was transformed by linear terminus(cut by PmeI) plasmid pShuttle-CMV-hp27mt into competent E. coli BJ5183 which contained adenovirus framework plasmid pAdeasy-1 to ensure that homologous recombination occurred. After correct identification of the transformant, it was digested by PacⅠ and transfected Ad293 cells, and packed into recombinant adenovirus Ad-hp27mt. Recombinant adenovirus was identified by polymerase chain reColon cancer cell line SW480 was infected with recombinant adenovirus Ad-p27mt, and expression of p27 protein was detected by Western blot.of expressed p27 protein after Ad-p27mt transfected colon cancer cell.novirus framework plasmid pAdeasy-1 with pShuttle-CMV-hp27mt, 30%combinant adenovirus DNA contained the target gene. Virus titer of the recancer cells, p27 was highly expressed in SW480 cells, as compared to low expression in non-transfected cells and Ad-LacZ transfected cells.CONCLUSION: Adenovirus, a type of gene transferring vector, can efficiently mediate p27 expression in tumor cells, which provides effective gene transfecting carrier for treating colorectal cancer with mutation p27 gene.

Objective To compare diabetes(pre-diabetes) prevalence and awareness between Province Shanxi and Fujian .Meth-ods Study data was from China National Diabetes and Metabolic Disorders Study 2007-2008 .5 926 individuals(Shanxi 3 254 ,Fu-jian 2 672) were included as study participants .Standardized questionnaire was used to obtain demographic and lifestyle data .All participants were administered a glucose tolerance test .Results Overall diabetes prevalence was 9 .6% .The diabetes prevalence in Shanxi was lower compared with Fujian(8 .2% vs .11 .3% ,P<0 .001) .Overall pre-diabetes was 15 .4% and no significant differ-ence was found between the two provinces .The awareness of diabetes and pre-diabetes in Shanxi was lower compared with Fujian , although both provinces were not satisfactory .Conclusion Regional differences were showed in diabetes (pre-diabetes) prevalence and awareness between Province Shanxi and Fujian .