Objective To further investigate inhibitory mechanism(s) of resveratrol on varicellazoster virus (VZV) in vitro with our previously generated reporter cell line MV9G.Methods Cell-free VZVs were directly inoculated onto MV9G cells (CFVs direct-infection) or cell-associated VZVs wereco-cultured with MV9G cells (CAVs co-culture) to activate expression of reporter gene firefly luciferase in MV9G cells.Resveratrol was added before or after virus infection,roles of resveratrolon direct inactivation,on viral attachment to and penetration into MV9G cells,on intracellular viral replication and its IC50,inhibitorytime points and reversibility were assayed by comparing the luciferase activities reduction by resveratrol.Thereductions of VZV IE62 mRNA copies and IE62-antibody positive cells by resveratrol were further assayed.Results ATPs contents of MV9G cells in the presence of resveratrol over 30.0 μg/ml were concentrationdependently reduced,the CD50 of which was around 60.3 μg/ml.CFVs were premixed with 25.0 μg/ml resveratrol andincubated at 37℃ waterbath for two hours and then directly inoculated onto MV9G cells,luciferases activated by resveratrol-treated CFVs were reduced to around half of the untreated controls.MV9G cells were pre-incubated with resveratrol at 37℃ for 2 h and then directly infected with CFVs at 37℃ for another 2 h,the CFVs-activated luciferase was concentration-dependently reduced,but no big change was observed in those pre-incubated at 4℃.MV9G cells were co-cultured with CAVs in the presence of resvertrolfor 72 h,the CAVs-activated luciferases were markedly reduced in a concentration-dependent manner,the IC50 of which was around 8.7 μgml.Resveratrol was added in CAVs co-culture at 1,3,6,9,12,24,30,and 36 h post infection,the CAVs-activated luciferase in those resveratrol was added at 3,6,9,12,and 24 h post infection were significantly higher than those of controls.Resveratrol was withdrawn from CAVs coculture media,the CAVs-activated luciferases after withdrawal were significantly higher than those before,especially in those withdrswn at 24 and 72 h post infection.The IE62 mRNA levels shown by cDNA copiesdetected with SYBR Green RT-PCR and IE62 positive cells shown by monoclonal anti-IE62 antibody of thevirus-infected cells treated with resveratrol were significantly reduced with increase of incubation time withresveratrol.Conclusion Resveratrol was cytotoxic to MV9G cells,and the maximum resistant concentrationon MV9G cells was around 30.0 μg/ml,the CD50 of which was around 60.3 μg/ml.Non-cytotoxic resveratrol partly inactivated CFVs,inhibited viral penetration into rather than attachment to MV9G cells.Resveratrol inhibited CAVs' intmcellular replication strongly but reversibly in a concentration-dependent manner,the IC50 of which was around 8.7 μ/ml.The inhibition of resveratrol on VZV in vitro might be through suppression of IE62 gene transcription and expression in the early stage of infection.

Objective To investigate mutations in immediate early ( IE) gene ORF62 of three var-icella vaccine Oka strains ( vOka ) including two strains produced in China and their parental Oka strain (pOka), and then to further elucidate its possible roles in attenuation mechanism by comparing their ORF 62 promoter sequences and its activities , ORF62 coding regions and its transactivities .Methods ORF62 pro-moter-reporter plasmids and ORF62-expressing plasmids of pOka and three vOka strains ( vOka-BK from Changchun BCHT Biotechnology Co ., vOka-SH from Shanghai Institute of Biological Product Co .Ltd., and vOka-GSK from GlaxoSmithKline plc, as control) were constructed, respectively.ORF62 promoter regions and coding regions of the four strains were sequenced and then compared with each other .Differences of ac-tivities of the ORF62 promoter, and transactivities of the ORF62-encoded IE62 upon immediate early (ORF4), early (ORF28) and late (ORF67) gene promoters between pOka and vOka strains were assayed with transient transfection technique .Results Compared with pOka strain , three vOka strains had a con-sistent T deletion mutation at site 110 050 in ORF62 promoters, which did not result in any change of tran-scription factor binding motif .However , activities of ORF62 promoters from three vOka strains were signifi-cantly lower than those of pOka strain .Three consistent substitution mutations were observed in ORF 62 cod-ing regions of three vOka strains and three new enzyme restriction sites including SmaⅠ, NaeⅠand BssHⅡwere generated, respectively.Transactivities of IE62 from three vOka strains upon ORF4, ORF28 and ORF67 promoters were significantly higher than those of pOka both in CV-1 and MeWo cells , except that vOka-SH IE62 showed significantly lower transactivities upon ORF 4 promoter than those of pOka strain in CV-1 cells.Conclusion Consistent T deletion mutation at site 110 050 in ORF62 promoters of three vOka strains might be responsible for the reduced promoter activities and the changes of IE 62 transactivities .How-ever , it seemed that cell types have no significant effect on ORF 62 promoter activity or IE 62 transactivity be-tween pOka and vOka strains .

Objective To investigate the arterial injury in vitro porcine kidney to different size of nephrostomy tracts. Methods The technique of percutaneous nephrostomy was applied to establish 11 groups of different size of nephrostomy tracts from 12 F to 32 F,with 40 tracts in every group.The technique of digital subtraction angiography (DSA) was used to inspect and analyze arterial injury. Results In the range from 12 F to 32 F,the damage of arterial injury increased with the size of the tract diameter.In groups of 20 F and 32 F,the number of nephrostomy tracts with serious arterial injury was 18 (18/40) and 30 (30/40) respectively,and the difference was statistically significant (P＜0.05).There was no statistical difference between groups 18 F、20 F、and 22 F. Conclusions The damage of renal arterial injury increased with the size of the nephrostomy tract.Atract of 20 F reduees arterial injury compared with a tract of 32 F.

Objective To retrospectively analyze treatment status quo of giant cell tumor of bone around the knee in several institutes,and to investigate risk factors affecting selection of surgical manners,tumor recurrence,and functional outcomes.Methods A total of 222 patients with giant cell tumor of bone around the knee confirmed by pathology,who had undergone surgical treatment in 5 institutes from March 2000 to May 2012,were enrolled in this study.There were 120 males and 102 females,with an average age of 35.5 years.The epidemiology,clinical and radiographic features and risk factors affecting selection of surgical manners,tumor recurrence,and functional outcomes were analyzed.Results Intralesional curettage was performed in 128 patients(57.6％),intralesional curettage combined with partial resection in 13 patients (5.8％),and marginal excision in 79 patients (35.5％).A total of 159 patients were followed up.The local recurrence rate was 19.0％ for patients treated with intralesional curettage,8.9％ for marginal excision and 0％ for intralesional curettage combined with partial resection,and the total local recurrence rate was 14.5％(23/159).The mean duration from primary operation to recurrence was 23.9 months.Univariate analysis indicated that surgical manner was the only factor affecting local recurrence.Pathological fracture,Campanacci grades,Enneking system as well as soft tissue mass had a significant influence on the selection of surgical manners.The mean MSTS score after intralesional curettage was obviously higher than that after marginal excision.Conclusion The surgical manner is the only factor affecting local recurrence and limb function.

On the basis of present hospital logistics management at home, the authors analyzed lean logistics management in hospitals and its practice in engineering works from the perspective of Changgung-featured management system, in terms of standard operating procedure, refined equipment maintenance, and routine skills training. It is held that refined logistics management can guarantee smooth operation and operating cost control for hospitals, which offers references for refined logistics management of hospitals in the country.