Objective To explore the effects of vitamin C and niacinamide on the growth and differentiation of human primary cultured keratinocytes.Methods Normal human foreskin was used in this study.The epidermis was separated enzymatically from the dermis by thermolysin,and keratinocytes were isolated from the epidermis by digestion with trypsin plus EDTA.The single keratinocytes were cultured with undedying NIH-3T3 cells as feeder cells in a complete medium supplied with 50 mg/L (vitamin C group),niacinamide of 400 μmol/L(niacinamide group)or vehicle(control group).Immunocytochemistry and immunodot blot were performed using monoclonal antibodies directed against C-myc,cyclin D1,filaggrin and involucrin.Results The colony number was highest in vitamin C group,followed by the control group and niacinamide group,and the colony morphology in vitamin C group was similar to that in the control group,but distinct from that in the niacinamide group.A significant increase was noticed in the expression of C-myc,cyclin D1,filaggrin and involucrin in vitamin C-treated keratinocytes compared with the control keratinocytes(all P＜0.05);however,in niacinamide-treated keratinocytes,the expression of filaggrin was significantly enhanced(P＜0.01),that of involucrin remained unchanged(P＞0.05),while that of C-myc was depressed(P＜0.05).Conclusions These results demonstrate that vitamin C has a favorable effect on both the growth and differentiation of human keratinocytes,while niacinamide seems to only promote the differentiation but attenuate the growth of human keratinocytes.

Objective To compare the difference in quercetin against oxidative stress response in mouse and in NIH-3T3 cells before and after H2O2 treatment,to explore the underlying mechanism for the quercetin antioxidant.Methods The cultured NIH-3T3 cells were randomly divided into 4 groups: quercetin(Q) pre-protective group(Qb) firstly treated with quercetin for 24 h followed by incubation with H2O2 for 30 min;post-protective group(Qa) treated with H2O2 for 30 min followed by incubation with quercetin for 24 h;H2O2 group(H2O2) after exposure to H2O2 for 30 min,incubated with DMEM medium and the control group(C) only cultured with DMEM medium.The survival rate and apoptotic rate were detected respectively with MTT and TUNEL in NIH-3T3 cell sus-pension samples.The expression of cyclin D1,PTEN,NF-?B,HSP-70,BCl-2,BAX and caspase-3 were examined with immunocytochemistry and immunoblotting.Besides,20 Wistar rats were divided into control group and experimental group,the latter was given with quercetin in the doze of 0.13 mmol/kg.The levels of T-AOC,SOD,GSH-Px,GSH,MDA,NOS and NO2-/NO3-were detected both in the cleaved NIH-3T3 cells and in the plasma from both experimental and control animals prior to and post-1 h,2 h and after 24 h.Results When the Qb group was compared with H2O2 or Qa group,the survival rate was higher and the apoptotic rate was lower.When the H2O2 group was compared with C group,the expression of cyclin D1、PTEN or BCl-2 was down-regulated;while that of BAX、HSP-70、NF-?B or caspase-3 was up-regulated;the level of T-AOC,SOD,GSH-Px or GSH was decreased;that of NOS、NO2-/NO3-or MDA enhanced in the cleft NIH-3T3 cells.When the plasma level of the anti-oxidative enzyme system prior to-compared with post-1h and 2h-treatment with Q,the level of T-AOC,SOD,GSH-Px and GSH,especially the former two,were higher;MDA,lower;NOS or NO2-/NO3-promoted.However,the above parameters basically became normal 24 h after treatment with Q.Conclusion Quercetin down-regulates the promoted expression of HSP70,NOS,NO2-/NO3-and NF-?B etc.in H2O2-treatment NIH-3T3 cells.Qb could reverse the H2O2 damage effects more markedly.Moreover,the quercetin exerts anti-oxidant protective effect through modulating the anti-oxidative enzyme system both in vivo and in vitro.However,based on the cell heterogeneity in none-or pre/post-H2O2-treatment state,a difference in quercetin antioxidant response is noted.

OBJECTIVE:To investigate the effects of doxorubicin perfusion therapy on therapeutic efficacy and related indexes in patients with superficial bladder cancer underwent transurethral resection of bladder tumor (TURBT).METHODS:Medical records of 96 patients with superficial bladder cancer were analyzed retrospectively and divided into observation group and control group according to drug use,with 48 cases in each group.Observation group was given perfusion therapy of 0.9％ Sodium choride solution 40 mL containing 20 mg doxorubicin 7 d after surgery.Control group was given TURBT combined with equal amount of 0.9％ Sodium chloride solution.Both groups received treatment for consecutive 10 months,once a week in first 8 weeks,later once a month.The recurrence rate and disease progression of 2 groups were observed and compared,and the levels of sICAM-1 and sVCAM-1,the levels of tumor marker DKK1 and VEGF were observed before and after treatment.The occurrence of ADR was recorded.RESULTS:After 24 months of treatment,remission rate and control rate of observation group were significantly higher than control group,with statistical significance (P＜0.05).There was no statistical significance in the progressive survival rate between 2 groups (P＞0.05).Before treatment,there was no statistical significance in the levels of sICAM-1、sVCAM-1、DKK1 and VEGF between 2 groups (P＞0.05);after treatment,the levels of sICAM-1、sVCAM-1 、DKK1 and VEGF in 2 groups were significantly lower than before treatment,the observation group was significantly lower than the control group,with statistical significance (P＜ 0.05).The recurrence rate,the incidence of frequent/urgent urination,hematuria and dysuria in observation group after treatment were significantly lower than control group,with statistical significance (P＜0.05).CONCLUSIONS:For patients with superficial bladder cancer underwent TURBT,doxorubicin perfusion therapy can significantly reduce recurrence rate,relieve tumor deterioration and reduce tumor activity with good safety.

Objective To explore the modulation effect of macrophage polarization and tumor microenvironment by colony-stimulating factor 1 receptor (CSF-1R) inhibitor administration in mice colon carcinoma tissue.Methods 24 mice were divided into normal control group,CAC group,CAC + GW2580 group.On day 64,the mice in CAC + GW2580 group were given GW2580 by daily gavage for a week.On 71 st day,the colon tissues were collected.The expression phenotype of macrophage polarization was detected by flow cytometry,the expression of macrophage-associated protein was detected by Western blot,and the expression level of various cytokine mRNAs were detected by PCR.Results The administration of CSF-1R inhibitors in CAC mice resulted in a significant reduction in the percentage of macrophages and M2 cells and a significant increase in M1 (P ＜ 0.05).The expression of M1-related proteins CD86 was up-regulated and the expression of M2-related proteins CD206 was down-regulated (P ＜0.05).Further more,the expression of M2-related IL-4,IL-10,Arg-1 were up-regulated (all P ＜ 0.05),and the expression of M1-related IL-12,iNOS were down-regulated (all P ＜ 0.05).Conclusion CSF-1R inhibitor can effectively modulate macrophage polarization from M2 to M1 in colon tumor and transform the tumor microenvironment from immunosuppression to immunostimulation,exerting tumor inhibition.