Objective:To screen the high performance,specific and nontoxic anti HBV antisense oligonucleotide fragment.Methods:Taking 2.2.15 cells as cell model,the antisense oligodeoxynucleotides(ASON)toward the HBV regulation genesenhancerⅡ(HBV EFⅡ)were designed and synthesized.The role played by ASON in inhibiting the secretion and expession of HBsAg and HBeAg by the host cells was detected by ELISA method.The effect of ASON on prolifezation and metabolism of the cells was detected by MTT method.Results:The inhibitory rate of HBsAg by ASON was 92% and 75%,respectively,indicating that itsdifference from that of noncomplementary sequence control group(inhibitory rate was 11%)had considerable statistical significance( P 0.05).Conclusion:ENⅡ is one of the most important target oriented sequencial selective regions of research on anti HBV nature of antisense oligonucleotide.

Objective To explore the clinical results of minimally invasive therapy for femoral intertrochanteric fractures with proximal femoral nail anti-rotation(PFNA) in 25 elderly patients.Methods A total of 25 patients with intertrochanteric fractures were treated by PFNA in our hospital from February 2007 to June 2008.The patients received close reduction and minimal invasive operation.Based upon the Tronzo-Evans classification,type Ⅰ fractures were diagnosed in 2 cases,type Ⅱ fractures in 2 cases,type Ⅲ in 16,and type Ⅳ in 5.Results The mean operation time in this series was 50.3 min(30-70 min),an the mean intraoperative blood loss was 100.2 ml(70-200 ml).All the 25 patients completed a follow-up of 3 to 12 months(average time 7.1 months),during which all the fractures healed in 10 to 14 weeks with a mean of 11 weeks,without occurrence of varus deformity,hardware loosening or hardware cutting.According to the Harris Hip Score,20 cases achieved excellent results,4 were good,and 1 was fair. Conclusions It is a reliable and ideal method to treat intertrochanteric fractures with PFNA in elderly patients,for the procedure is convenient and minimally invasive,resulting in firm fixation,less blood loss,short operation time,and high healing rate.

Objective To explore the mechanism of immune tolerance in chronic B hepatitis, the effects of HBV infection on the functions of macrophages and related mechanism of signal transduction by transfection in vitro were studied. Methods Peritoneal macrophages of mice were isolated regularly and cultured, transfected transiently with pcDNA3-HBV or pcDNA3 plasmid DNA and cultured under the stimulation of LPS. After 72 h, RT-PCR was performed to detect the expression of PreS1, TNF-? or IL-1? mRNA. To detect the expression of NF-?B RelA protein by FCM, and the level of nitric oxide in cultural supernatant was measured with Griess reaction. Results After being transfected with pcDNA3-HBV,peritoneal macrophages had the expression of PreS1 mRNA, but have lower level of TNF-?、IL-1? mRNA and transcriptional factor NF-?B, compared with pcDNA3-transfected control group; the level of nitric oxide in pcDNA3-HBV group was also decreased. Conclusions Transient transfection of pcDNA3-HBV could decrease the function of macrophages directly by inhibiting NF-?B activity and effector molecules production, which may be one of the mechanisms of immune tolerance in chronic B hepatitis.

本文在“国际残疾分类”的基础上，围绕康复诊断名称的标准化，进行了初步探讨，力求使之适合于临床康复的应用及康复病案管理。

0.05).The C myc and N ras protein expression of 2.2.15 were reduced after ASONs were used 3 days.There was a significant difference between ASON group and control group ( P

Objective To investigate the possibility of transfection of adipose-derived mesenchymal stem cells (ADMSCs) into the insulin-secreting cells in vitro,and assay the secretion of insulin of ADMSCs in high and low glucose environment.Methods The ADMSCs that untransfected were in the control group,the ADMSCs that contained PcDNA3.1 were in the vacant vector group,and the ADMSCs that contained PcDNA3.1-hINS were in the recombinant vector group.After transfection,the recombinant vector group were sub-divided into the 1,6,12,18 days groups.According to the concentrations of glucose,the recombinant vector 18 days group were divided into the high glucose group and low glucose group.Human insulin gene was amplified by RT-PCR,and the eukaryotic expression recombinant vector PcDNA3.1-hINS that contained the human insulin gene was constructed.The ADMSCs were obtained by digestion and centrifugation,and then underwent flow cytometry for identification.The transcription of insulin DNA was assayed by RT-PCR,and the levels of insulin were assayed by ELISA.Glucose test was done in the recombinant vector 18 days group.The measurement data was shown in the format of (x) ± s,the measurement data in multiple groups were compared by randomized analysis of variance,and the comparison among groups was performed by the t test.ResuIts The expressions of CD44,CD90,CD106 were positive,and the expressions of CD34,CD45 and CD11b were negative.No insulin DNA transcription was detected in the control group and vacant vector group.The levels of insulin secreted were (4.7 ± 0.8) mIU/L,(8.8 ± 0.5) mIU/L,(8.9 ± 0.8)mIU/L,(8.6 ± 0.6)mIU/L in the recombinant vector 1,6,12,18 days group,which were significantly higher than (1.3 ± 0.6) mIU/L in the control group and (1.7 ± 0.8) mIU/L in the vacant vector group (t =10.09,32.64,22.20,55.53 ; 9.23,27.56,19.43,51.25,P ＜ 0.05).There were significant differences in the levels of insulin secreted between the recombinant vector 1 day group and the recombinant vector 6,12,18 days groups (t =12.77,12.26,13.93,P ＜0.05).There were no significant difference in the levels of insulin secreted between the recombinant vector 6,12,18 days groups (F =45.67,P ＞ 0.05).There was a significant difference in the level of insulin secreted between the high glucose group and the low glucose group (t =2.03,P ＜ 0.05).The result of the glucose stimulation test was negative.Conclusion The ADMSCs are transfected into insulinsecreting cells in vitro successfully,and the secretion of insulin is stable.Although the secretion of insulin can't change in line with the concentration of glucose,it is a new seed cell for the treatment of diabetes with stem cells.

AIM: To explore the effect of HBV transfection on apoptosis induced by TWEAK and the regulatory effect of IFN-γ on the apoptosis process.METHODS: BEL-7402-pCDNA3/1.1HBV hepatoma cell line was used as cell model in this experiment, in which BEL-7402 was stably transfected with pCDNA3/1.1HBV and its corresponding empty vector BEL-7402-pCDNA3 was used as control. The effect of HBV transfection on TNF-like weak inducer of apoptosis(TWEAK) induced hepatoma cell apoptosis was determined by CCK-8. The apoptosis before and after HBV transfection induced by TWEAK and the effect of IFN-γ on apoptosis induced by TWEAK in HBV transfected hepatoma cells were detected by TUNEL and caspase 3 activity detection kit.RESULTS: BEL-7402-pCDNA3/1.1HBV and BEL-7402-pCDNA3 hepatoma cell line were successfully established and identified. Cell viability of hepatoma cells was increased after HBV transfection. The apoptosis rate was much higher after HBV transfection compared to the empty vector control and empty cell control. The apoptosis rate induced by TWEAK was much higher in BEL-7402-pCDNA3/1.1HBV cells when exposed to IFN-γ.CONCLUSION: After HBV infection, TWEAK might be involved in the clearance of HBV infected hepatocyte by apoptosis and inflammatory factor such as IFN-γ.

Objective:To synthesize disubstituted pyridine and I-substituted propynyl carbamates and study their antimicrobial activities, searching for more potent and less toxic antimicrobial agents.Methods: The title compounds were synthesized through the process of electrophilic and nucleophilic substitution. Antimicrobial test in vitro was determined with 13 kinds of common mildews and bacteria(Aspergullus niger, aspergillus flavus, Aspergillus versicolor,Trichoderma viride, Paecilomium varioti Bainier, Chaetomium globsum, Penicillium citrinum, Cladochytrium clodospoium, Escherichia colo, Staphylococcus aureus, Pseudomonas fluorescens Bacillus fluorescens, and Bacillus megatherium). The chemical structures of all compounds were determined by IR, 1 HNMR and elementary analysis.Results: Seven disubstituted pyridine and I-substituted propynyl carbamates obtained were firstly reported. All compounds showed antimicrobial activity, especially compound 3e, who had more potent activity compared with the that of 3-iodo-2-propynyl-butyl- carbamate (IPBC). MIC of the compound 3e was between 2?10 -6 g/ml to 30?10 -6 g/ml . Conclusion: Compound 3e has the best antimicrobial activity and should be futher studied.

Objective To explore the role of TRAIL receptors expression on peripheral blood mononuclear cell (PBMC) in the apoptosis of PBMC,and the relation of the expression of TRAIL receptors with the severity of liver damage in chronic hepatitis B patients.Methods The expressions of DR4,DRS,DcR1 and DcR2 in 55 patients and 30 cases of control were assayed by RT-RCR and floweytometery.The relationship of the expression of TRAIL receptors with the severity of liver damage were analized.Results The level of DcR1 in the PBMC of chronic hepatitis B patients was much lower than that of control group (P＜0.05).There was close relation between the expression of DcR1 and liver damage (P＜0.05).The level of DcR1 was negatively correlated with ALT but positively correlated with serum albumin.Conclusion The expression level of DcR1 on PBMC in chronic hepatitis B patients was down-regulated,which may contribute to the increased apoptosis of PBMC in chronic B hepatitis patients.The expression of DcR1 can reflect the degree of liver injury in chronic hepatitis B patients to some degree.

0.05). The mainly side-effects were myelosuppression, nausea, and vomiting. Conclusions:Docetaxel/cisplatin and gemcitabine/cisplatin regimens were well toleranced in advanced NSCLC patients with long term survival.