Objective:To investigate the interfering effect of siRNA on the expression of VEGF in Hela cells.Methods:Three pairs of siRNA were designed according to the 1-5 extrons sequence of VEGF and synthesized by T7 RNA ploymerase transcription in vitro. To evaluate the inhibition activity of siRNA, Hela cells were transfected via siPORT Lipid. The interfering effect of siRNAs in hRPE cells was examined by semi-quantitative RT-PCR and immunofluorescence technique.Results:The results showed that the three pairs of siRNA could effectively inhibit gene expression of VEGF in Hela cells with the rates of 88.7%, 94.2% and 80.3%. But the 1-base mismatched siRNA and ssRNA(+) didn′t show significant interfering effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect(5-10 pmol).Conclusion:The siRNAs synthesized by T7 RNA ploymerase in vitro could effectively and specifically interfere the expression of VEGF in Hela cells, providing a novel approach for gene therapy of tumor.

Objective To investigate the effect of Helicobacter pylori infection on cell proliferation and apoptosis of gastric cancer cell line SGC 7901. Method SGC 7901 cells were incubated with 1 × 108,5 × 107,1 × 107,5 × 106 cfu / ml concentration gradient of the standard HP NCTC 11637 strains in vitro. Morphological changes of cells were observed at 24, 48, 72 hour, respectively. Cell proliferation inhibition rate were detected with MTT assay. Apoptosis were observed by flow eytometry and TUNEL analysis, mRNA expression of survivin were detected by RT-PCR, and survivin expression were determined by western blot. Results Cell proliferation inhibition rate was 54.5%, 58.9%, 67.6%, 72.9% with 1 × 108,5 × 107, 1 × 107,5 × 106 cfu / ml concentration gradient of the HP bacteria on the role of SGC 7901 cells at 72h, respectively. The rate of apoptosis after 72h detected by flow cytometry and TUNEL with HP concentration gradient of different bacteria were 42.51%, 45.67%,48.57%, 49.51% and 54.61%, 51.26%, 59.41%, 62.46.% Helieobacter pylori could significantly reduce mRNA and protein expression of survivin. These effects were strengthened as the concentration of Helicobacter pylori increased and the time extended. Conclusion H. Pylori infection may be through reducing the expression of survivin to inhibit cell proliferation and pro-mote apoptosis of SGC 7901 in vitro. Moreover, this effect was positive related with time and dose.

Objective_To investigate whether hUCMSCs undergo malignant transformation when exposed to MCF-7B breast cancer microenvironment and whether the abnormal activation and over expression of STAT3 play an impor-tant role in this transformation.Methods_The experiment was divided into three groups:blank group ( hUCMSCs were separately cultured),experimental group(hUCMSCs were indirectly co-cultured with MCF-7B breast cancer cells),positive control group(MCF-7B breast cancer cells were separately cultured).Morphology of cells was detec-ted by invertedmicroscope.Cell cycle was detected by flow cytometry.The mRNA expression of STAT3, c-Myc and Bcl-xL was tested by real-time PCR.The protein expression and location of p-STAT3,c-Myc and Bcl-xL were detected by immunofluorescence.The protein expressions of p-STAT3,STAT3,c-Myc and Bcl-xL were also meas-ured by Western blot.Results_The experimental group cells showed typical morphology of the tumor cells.The cells proportion of experiment group in G1 phase was significantly lower than that of the blank group(P<0.05),but which in S and G2 phase were significantly higher than those of the blank group(P<0.05).The mRNA expression levels of STAT3 ,c-Myc and Bcl-xL in experimental group was significantly higher than those in blank group ( P<0.05).p-STAT3,STAT3,c-Myc and Bcl-xL protein were significantly higher than those of the blank group(P<0.05),and they were mainly located in the nuclei.The protein expression of STATS also showed significant changes in experimental group.Conclusions_hUCMSCs trends to malignant transformations when exposed to MCF-7B breast cancer microenvironment.The abnormal activation and over expression of STAT3 are of important factors leading to the malignant transformation of hUCMSCs.

Objective To investigate the suppression to 2-glycoprotein-1-specific B cell response by human recombinant 2-glycoprotein-1 domain Ⅰ dimer(rh?2-GP1-DⅠ2) in rh?2-GP1 immunized mice.Methods The effects of treatment using rh?2-GP1-DⅠ2 on titer and ratio of anti-?2-GP1 were analyzed by solid phase ELISA.The number of splenic ?2-GP1-specific antibody-forming cells(AFC) was counted by ELISPOT.Results The levels of anti-?2-GP1 from rh?2-GP1 immunized mice treated with rh?2-GP1-DⅠ2 were significantly decreased compared with those in negative controls(P

Protein phosphorylation is one of the most common post-translational modifications (PTMs)in various organisms,which plays critical roles in the regulation of intracellular biological processes,such as cell prolifera-tion,signal transduction,metabolismis and tumorigenesis.However,the low abundance of phosphoprotein in the biological systems poses significant challenges of current analytical techniques.In order to further understand the phosphoproteomics,the roles of phosphorylated proteins in life process,discovery of biomarkers,diagnosis and treatment of disease,enrichment strategies of high efficiency have been developed,including the design of new nanomaterials and combination of a variety of analytical methods,et al.In this paper,we reviewed the develop-ment of enrichment strategies for phosphoproteomics and application of phosphoproteomics in disease.

Objective To discuss clinical effects and indications of minimally invasive tension band technique in treatment of transverse patella fractures.Methods Thirty-eight patients with transverse patella fracture (close or slightly polluted open ones) were treated with minimally invasive tension band technique from July 1997 to June 2005 in our department.Twenty-six were fixated with percutaneous Kirschner wire tension hand and 12 with percutaneous cannulated screw tension band.The reduction,movement range and function of the knee were analyzed.Results Surgeries went on uneventfully.All the cases were followed up from six months to 6.5 years (average,32 months).Fracture healing time lasted from 6 to 12 weeks,averaging 7.7 weeks.No such compli- cations as traumatic arthritis occurred.Reduction was excellent in 21 cases,good in 11 and fair in six,with the good-to-excellent rate being 84.2%.All the affected knee joints could move in normal range.The functional re- covery was assessed as excellent in 34 cases and good in four.Conclusions Internal fixation with minimally invasive tension band technique is good for treatment of transverse patella fractures.It is chiefly fit for close or slightly polluted open fractures with limited wound,and can also be used for the comminuted fractures if the bone fragments are no more than three and the fracture site is suitable.

Objective To study the effects of Liensinine on apoptosis of 5637 cells, and its mechanism thereof. Meth?ods CCK-8 method and the colony formation test were used to detect cell viabilities, and then inhibition rates were calcu?lated. Flow cytometry was used to detect the effects of Liensinine on apoptosis of 5637 cells. Western blot assay was used to detect Caspase-7 protein expression. Results CCK-8 assay and colony formation test indicated that Liensinine inhibited the cell proliferation significantly. Results of flow cytometry indicated that Liensinine induced early apoptosis of 5637 cells. Western blot assay showed that Liensinine improved the expression of Caspase-7 and enhanced the activation of Caspase-7 in 5637 cells. Conclusion Liensinine could inhibit the proliferation of 5637 cells, induce early apoptosis, which may be re?lated with the enhanced expression of Caspase-7 and its activation.

Alzheimer disease (AD) is a common neurodegenerative disease. Drosophila has been regard as one of the ideal models for Alzheimer because of its unique advantage on genetic manipulation. AD transgenic drosophila models not only help to elucidate the pathogenesis of Alzheimer disease, but also provide potential screening models for drugs to treat the disease. In this review, we summarize the recent research progress using AD transgenic drosophila.
Alzheimer Disease ; etiology ; genetics ; metabolism ; Amyloid beta-Peptides ; metabolism ; Animals ; Animals, Genetically Modified ; genetics ; Disease Models, Animal ; Drosophila ; genetics ; Drug Evaluation, Preclinical ; Gene Transfer Techniques ; Humans ; Phosphorylation ; tau Proteins ; metabolism

OBJECTIVETo investigate the efficacy and safety of circumferential pulmonary vein ostial isolation guided by EnSite NavX three-dimensional electrophysiological mapping in patients with atrial fibrillation (AF).

METHODSThirty-eight patients with drug refractory paroxysmal or persistent AF underwent circumferential pulmonary vein ostial isolation and were followed up to investigate the efficacy and safety of the treatment.

RESULTSAll cases reached the endpoint of the ablation, and both sides of the pulmonary vein were completely isolated, with an average procedure time of 200.4-/+37.0 min, X-ray exposure time of 54.7-/+9.7 min, and three-dimensional left atrial geometry reconstruction time of 27.5-/+7.5 min. During the follow-up for 9-/+3 months, the success rate of initial ablation was 89.5%, and the incidence of procedure-related complications were 7.9%.

CONCLUSIONSCircumferential pulmonary vein ostial isolation guided by EnSite NavX three-dimensional electrophysiological mapping can be effective and safe for AF treatment.

Adult ; Aged ; Atrial Fibrillation ; surgery ; Catheter Ablation ; methods ; Electrophysiologic Techniques, Cardiac ; methods ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Veins ; surgery ; Treatment Outcome