Objective To study the effects of miR?21 on the proliferation,apoptosis and MMP2 expression of HeLa cells. Methods The miR?21 or the miRNA scramble control was transfected into Hela cells. The cell proliferation was detected by the Celltiter?GloTM assay 72 h after the transfection,and the apoptosis was evaluated by the Caspase3/7 Glo? Kit. The MMP2 RNA expression was quantified by quantitative real time PCR. Results The proliferation of HeLa cells transfected with miR?21 was significantly increased compared to that of the cells transfected with miRNA scramble control. The caspase 3/7 activity in HeLa cells transfected with miR?21 was downregulated compared to that of the cells transfect?ed with miRNA negative control. The MMP2 RNA expression in HeLa cells transfected with miR?21 was increased significantly compared to the cells transfected with miRNA negative control. Conclusion miR?21 can significantly promote the proliferation of HeLa cells. In addition,it ex?hibits the anti?apoptosis effect in HeLa cells. The transfection of miR?21 significantly increased the MMP2 RNA expression,which suggests that miR?21 may promote the tumor invasion and might be a therapeutic intervention target.

Aim To investigate the effect of triptolide ( TP) on human colorectal cancer HCT116 cell prolif-eration and explore the potential mechanism of TP in treating colon cancer. Methods MTT assay was used for estimating the survival rates of HCT116 cells ex-posed to different concentrations and different duration time of TP. Western blot ( WB ) was used for testing the expression of LC3-Ⅱ. MDC staining was employed to detect cell autophagy. Flow cytometry ( FCM) was applied to test the effects of TP on cell apoptosis. Re-sults TP could inhibit cell proliferation in HCT116 cells. The expression of LC3-Ⅱwas enhanced with the increase of the concentration of TP after exposed to dif-ferent concentrations of TP for 48 h and the duration time of TP after exposed to 40 nmol·L-1 TP,and the number of acid autophagic vacuoles in HCT116 cells had increased, which was observed by fluorescence mi-croscope. The HCT116 cells apoptotic rates in TP group had decreased compared to control group and de-creased significantly compared to TP +3-MA group. The HCT116 cells apoptotic rates in TP group had in-creased compared to control group and increased signif-icantly compared to TP+RAPA group. Conclusion TP could inhibit the proliferation of human colon canc-er HCT116 cells and induce apoptosis in HCT116 cells, which indicates that synergy may exist between autophagy and apoptosis.

OBJECTIVEThe survival rate of cadaveric renal transplant in children has been improved following the development of transplantation technology and the application of immunosuppressive agents. In this study, the prognosis of renal transplantation, operative procedure and immunosuppressive agents administration in 21 children with end-stage renal disease (ESRD) were analyzed.

METHODSFrom January 1985 to December 2001, 21 patients (9 males and 12 females with a mean age of 14 +/- 2 yr, mean body weight of 33.4 kg and mean height of 136.5 cm) received renal transplantation because of ESRD were enrolled in the study. The patients with an average GFR of 8.28 ml/min were managed with dialysis for 13.4 months in average pro-transplantation. All cadaveric kidneys were from adults, which included 1 donor with one HLA mismatch, 3 with two mismatches, 5 with three mismatches and 4 with four mismatches. All transplantations were performed with anastomoses of the adults' renal artery and vein to the children's iliac externa artery and iliac externa vein. Biological inducement therapy was given in 4 cases. At the first 3 - 5 days post-transplantation, methylprednisolone was administered [7 mg/(kg.d)]. All patients received baseline diploid or triple immunosuppression therapy of cyclosporin A [6 - 8 mg/(kg.d)] or FK506 [0.18 - 0.25 mg/(kg.d)], mycophenolate mofetil [MMF 10 - 15 mg/(kg.d)] or azathioprine [1 - 3 mg/(kg.d)] and prednisone [0.4 - 0.6 mg/(kg.d)]. High-dose methylprednisolone [10 mg/(kg.d)] was administered to control the acute rejection.

RESULTSThe renal function of patients was restored 5.6 days in average after transplantations. The 1st, 3rd and 5th year survival rates of recipient/graft were 95.2%/95.2%, 86.7%/73.3% and 72.7%/63.6%, respectively. One case had super-acute renal rejection, 5 cases had acute rejection, 3 cases had delayed graft function and 3 patients died. The longest survival time was 12 years. The major complications included hypertension (47.6%), diabetes (19.4%), infection (19.4%) and drug-induced hepatic injury (14.2%). Catch-up growth was seen in most of the pediatric recipients.

CONCLUSIONRenal transplantation is the most ideal method to treat children with ESRD, and the growth of the pediatric patients will be improved after transplantation. Adult donor kidneys adapt to the school age patient. And the protocol of immunosuppressive therapy (prednisone plus MMF and FK506) should be applied.

Adolescent ; Adult ; Child ; Female ; Histocompatibility Testing ; Humans ; Kidney Failure, Chronic ; mortality ; therapy ; Kidney Transplantation ; Male ; Postoperative Care ; Preoperative Care ; Prognosis ; Survival Rate ; Time Factors

Objective:To evaluate immune response of murine peritoneal macrophage challenging by methicillin-resistant S.aureus(MRSA)after pretreatment with Pam3Csk4(TLR2 agonist).Methods: Murine peritoneal macrophage was pretreated with Pam3Csk4(1 μg/ml).Following pretreatment 12 h later,heat-killed MRSA( HK-MRSA) was added and incubated for another 2 or 6 hours.The protein and mRNA level of TNF-α, IL-6 and IL-1 were determined by ELISA and Q-PCR, respectively.To estimate phagocytosis of macrophage,HK-MRSA/MSSA labeled with FITC( FITC-HK-MRSA/MSSA) were added to well and incubated for 30 min.After washing 5 times with PBS,intracellular FITC-HK-MRSA was detected by flow cytometry.To estimate antimicrobal activity of macrophage,live MRSA and MSSA were added to well and incubated at indication time,the CFU of s.aureus was estimated via a 10-fold serial dilution on agar media.cDNA was further quantitative assessed using primers for mouse FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 by Q-PCR .Results: Compared with saline-pretreated cell, the protein and mRNA level of TNF-α, IL-6 and IL-1 were markely reduced, respectively.However, both the phagocytosis and antimicrobal activity to S.aureus were significantly increased in macrophages pretreated with Pam3Csk4.Further study found that the macrophages had higher FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 expression at 6 h and 12 h post-stimulation Pam3Csk4.Conclusion: The results suggest that Pam3Csk4 could activate murine antimicrobal activity of peritoneal macrophage challenging by methicillin-resistant Saureus via increasing opsonophagocytosis in depended antibodies, complements manners.The results suggest Pam3Csk4 probably be a novel immunotherapy candidate against MRSA.