Objective: To study mouse Id3(mId3) protein expression in WEHI-231 B lymphoma cells after CD40 ligand(CD40L) treatment. Methods: The mId3 cDNA was amplified from total RNA of WEHI-231 cells by RT-PCR and introduced into the p3FLAG-CMV-10 vector.The 3FLAG-Id3 fusion gene was amplified from p3FLAG-Id3-CMV-10 vector and subcloned into retroviral vector pMX-CITE/IRES-EGFP.Recombinant retrovirus was produced by transient transfection of the Phoenix-ecotropic packaging cell line with pMX-3FLAG-Id3-EGFP vector using FuGene-6 transfection reagent.(WEHI)-231 cells were transduced with recombinant retrovirus.EGFP expression was monitored by flow cytometry.Two cell lines stably expressing 3FLAG-Id3 fusion protein were generated. Results: The proportion of EGFP-positive cells in the population transduced with pMX-3FLAG-EGFP remained constant up to 2 days after transduction,whereas the proportion in the population of pMX-3FLAG-Id3-EGFP-transduced cells decreased with time.After coculturing with NIH3T3 cells or medium alone,the proportion of EGFP-positive cells in pMX-3FLAG-Id3-EGFP-transduced WEHI-231 cells decreased with time,whereas after coculturing with CD40L-NIH3T3 cells,the proportion of EGFP-positive cells in the transduced cells remained constant up to 2 days,suggesting that CD40L could rescue WEHI-231 from Id3 overexpressioninduced growth arrest.After coculturing with CD40L/NIH3T3 or NIH3T3 cells,3FLAG-Id3 fusion protein stably expressing WEHI-231 cells were harvested,lysed,and immunoblotted by using anti-FLAG antibody.The results showed that treatment via CD40 ligation induced significant downregulation of Id3 expression in pMX3FLAG-Id3-EGFP tansduced WEHI 231 cells. Conclusion:CD40 ligation pvevents the Id3 induced cell growth avvest by the dousn-regulation of Id3 expression.

Objective To investigate the effects of inferior vena cava (IVC) filter placement for the prevention of fatal pulmonary embolism and to discuss the management of complications related to the filter placement. Methods Seventy patients with proved deep vein thrombosis of lower extremity underwent inferior vena cava filter placement. A total of 72 IVC filters were implanted, which included 20 Trap Ease filters, 31Vena Tech filters, 13 retrievable OptEase~(TM) filters and 8 Tempo Ⅱ filters. One filter was deployed above the orifice of renal vein and the remaining 71 were deployed below the orifice of renal vein. Results All the patients were followed up for 8-72 months after the procedure. During the follow-up period no fatal pulmonary embolism occurred except that some complications related to the filter placement occurred in 6 cases.Conclusion Inferior vena cava filter placement can effectively prevent the occurrence of pulmonary embolism. Of course, this treatment should be strictly applied according to the indications.

Dendritic cells(DCs) are known as the most efficient antigen-presenting cells that are potent stimulators of both B and T cell immune responses.The biological characteristics and culture in vitro of DCs,preparation of DC vaccines,experimental and clinical studies in tumor immunotherapy by DC vaccines are reviewed in this article.

Objective To investigate the expression and significance of cyclin E in rectum carcinoma.Methods The expression of cyclin E was examined by immunohistochemical techniques in 42 cases of rectum carcinoma.Laboratory data were then analyzed statistically together with the related clinical and pathological data.Results The positive expression rate of cyclin E in rectum carcinoma was 66.7％(28/42).There was no significant association between cyclin E and gender,age,histological grade,pTNM stage,metastasis of lymph node (P ＞ 0.05).Conclusions The expression of cyclin E in rectum carcinoma is higher,and it may show highly associated with the occurrence and development of the rectum carcinoma.Cyclin E has no significant association with age,gender,histological grade,pTNM stage,metastasis of lymph

Objective: To observe the clinical effect of integrated traditional and western medicine in treating chronic idiopathic thrombocytopenic purpura (CITP) elderly patients. Methods: Sixty patients were divided randomly into the treatment group (n=30) and the control group (n=30), the former was treated with danazol combining with prednisone and Chinese drugs, while the latter was treated with VCR. Results: The total effective rate of the treatment group was better than that of the control group, the difference between those two groups was significant (P

[Objective]To observe the clinical treatment of calcaneus fracture with poking reduction and artificial bone grafting technique.[Method]From 1999.6 to 2005.5,60 cases with recent calcaneus fracture,including 46 feet in 38 males and 24 feet in 22 females with falling injure;aged from 20 to 65,were equally divided into two groups,including the observed group treated with poking reduction and artificial bone grafting and the control group with steel needle pry-poking reposition.[Result]During followed up for 6-60 months,average 20.7months,it was notable different in Maryland foot score,and the same in Bhler′,Gissane′ and Perie′ angle in X-ray in post-operation and pre-operative between observed group and the control.[Conclusion]It showed us that the technique of poking reduction and artificial bone grafting was liable to operate and could provide exactly efficiency.

OBJECTIVE: To summarize the development of researches on sildenafil for treating cardiovascular patients with erectile dysfunction, so as to explore the safety of applying sildenafil and efficacy of treating related cardiovascular diseases.DATA SOURCES: The relevant articles to sildenafil between October 1999 and February 2006 were computer-searched in PubMed databasewith the key words of "sildenafil, erectile dysfunction, sexuality, cardiovascular disease" in English. Meanwhile, Medscape database was scanned with computer to identify relevant articles with key words of "sildenafil, cardiovascular disease". STUDY SELECTION: After the primary searching, literatures on relationship between sildenafil and cardiovascular disease were searched in full-text, while articles about erectile dysfunction but had nothing to do with cardiovascular disease were excluded. The repetitive experimental studies were also deleted. DATA EXTRACTION: A total of 55 articles were collected, and 22 of them were in accordance with the inclusion criteria and their full-texts were carefully looked through. DATA SYNTHESIS: The selected data were classified according to the relationship between erectile dysfunction and cardiovascular disease, effect of sildenafil on erectile dysfunction of patients with heart disease and rec ommendation of the Princeton conference. CONCLUSION: It is demonstrated in many literatures that the potential risk of sildenafil triggering cardiovascular disease is relatively small. Cardiac patients need not promote huge concerns when resorting to sildenafil for erectile dysfunction treatment. However, the protective cardiac effects of sildenafil should be further investigated.

Objective To study the effects of inhibiting forkhead transcription factor O1 (FoxO1)expression by small interference RNA (siRNA) on the glucose utilization of insulin resistant HepG-2 cell line and the mechanism.Methods FoxO1 gene-targeted siRNA vector,which carried a red fluorescence,was constructed and then confirmed by DNA sequencing.HepG-2 cells were induced to a status of insulin resistance by being exposed to 10-6 mol/L insulin for 24 h.The study included 4 groups: HepG-2 cell group cultured with normal medium (group A) ; insulin resistant HepG-2 cell group (group B) ; insulin resistant HepG-2 cell group into which FoxO1 siRNA vector was transfected (group C) ; insulin resistant HepG-2 cell group into which Lipofectamine2000 was added (group D).The transfection efficiency could be estimated by observing the expression of red fluorescence.The expression of FoxO1 mRNA was analyzed by RT-PCR.The expressions of IRS-2 and tyrosine phosphorylation were detected by Western blot and immunoprecipitation.Results FoxO1 gene-targeted siRNA vector was built successfully.The expression of red fluorescence was the strongest at 48 h after transfection.Compared with group A,the glucose consumption and the expression of IRS-2 tyrosine phosphorylation of group B were decreased (P<0.01),and expression of FoxO1 mRNA was increased (P<0.05) in group B.There was no difference in the expressions of IRS-2 protein between A and B groups.Compared with group B,the expression of FoxOl mRNA was decreased (P < 0.01),and the glucose consumption and expression of IRS-2 tyrosine phosphorylation were significantly increased (P<0.05) in group C.There was no difference between group D and group B in glucose consumption, FoxO1 mRNA, IRS-2 protein and IRS-2 tyrosine phosphorylation expressions.Conclusions Inhibiting the expression of FoxO1 gene in insulin resistant HepG-2 cells seems to improve insulin sensitivity by feedback regulating the IRS-2 tyrosine phosphorylation expressions.

Objective To study the effects and mechanisms of resveratrol (Resv) on the expression of adiponectin receptors ( AdipoR1 and AdipoR2 ) in rat mesangial cells (MCs) cultured in intermittent high glucose medium.Methods The MCs cultured in normal glucose ( 5.6 mmol/L) medium were transfected by the plasmid vector of the FoxO1 short hairpin RNAs ( FoxO1 shRNA ),and then cultured in intermittent high glucose medium (5.6mmol/L or 30 mmol/L,alternately every 8 hours ) and resveratrol ( Resv 20 μmol/L).The MCs cultured in normal glucose (5.6 mmol/L) medium served as normal glucose control group (NG),and the MCs exposed to iutermittent high glucose medium were divided into five groups:intermittent high glucose group (IHG),IHG+Resv group,IHG+FoxO1 shRNA group,IHG+ Resv + FoxO1 shRNA group,IHG + Negative control group ( shNC ),each group was cultured for 72 h.The level of reactive oxygen species (ROS) was assessed by Fluorescence microplate reader.The mRNA levels of Sirt1,Foxo1,AdipoR1,and AdipoR2 were assessed by RT-RCR.The protein levels of FoxO1,phosphorylation FoxO1 ( p-FoxO1 ),AdipoR1,and AdipoR2 were assessed by Western blotting.Results ( 1 )Compared with NG,intermittent high glucose significantly decreased the mRNA expression of Sirt1,but markedly increased the level of p-FoxO1.Furthermore,the mRNA and protein expression levels of AdipoR1 were obviously decreased while the level of ROS was enhanced in IHG ( all P＜0.05 ).(2) Compared with IHG,the mRNA and protein expression levels of FoxO1 and AdipoR1 were inhibited,but the level of ROS was enhanced in group IHG+FoxO1 shRNA ( all P＜0.05 ).( 3 ) Compared with IHG,after treating with Resv,the mRNA expression level of Sirtl was increased,whereas the level of p-FoxO1 was decreased.Moreover,the mRNA and protein levels of AdipoR1 was increased while the level of ROS was lowered in group IHG+Resv (all P＜0.05 ).(4) Compared with group IHG+Resv,the mRNA and protein levels of FoxO1 and AdipoR1 were inhibited,but the level of ROS was increased in group IHG+Resv+FoxO1 shRNA (all P＜O.05).(5) In addition,the mRNA and protein expression level of AdipoR2 showed no significant difference among these groups ( P＞0.05 ).Conclusion ( 1 ) Resveratrol significantly increases the expression of AdipoR1 in MCs induced by intermittent high glucose.( 2 ) FoxO1 plays an important role in regulating the expression of AdipoR1 by resveratrol.

Objective To determine the plasma concentration of cytarabine(Ara-C) in children with leukemia and obtain dynamics parameters, and investigate the relationship between the parameters and clinical effect in order to provide the basis for optimization of Ara-C application. Methods Using highperformance liquid chromatogram (HPLC) to determine the plasma concentration of Ara-C, its metabolite Ara-U and infusion rate in 37 children with acute leukemia, their therapeutic reaction, remission, treatment-related infection, side-effect and long-term treatment effect were analyzed in statistic. Results Ara-C by 1～2 g/m2 intravenous drop infusion for 2 hours, the peak plasma concentration time was 2 h and peak concentration were (14.37-84.44)μmol/L, and the median was (41.42±22.80)μmol/L. The median infusion rate was 869.57at 30 minutes after Ara-C drip completion, its average level was (253.40±81.49) μmol/L, over six-times than Ara-C peak concentration. The median continuous complete remission time in 37 children was 29.8 months (5.0～53.1 months), 3y-DFS was (90.63±5.15)%. The therapy-related infection rate was 56.8 %(21/37),including three children (8.1 %) suffered from severe infection, but there was no therapy-related death and no children were off the protocol due to poor tolerance. Conclusion As post-remission treatment, high-dose Ara-C would not cause cumulation in vivo in children with acute leukemia and side-effect were slight. Ara-C could improve the long-term continuous complete remission rate and clinical cure rate for children with leukemia. Therefore, it was worth to apply in clinical.