Objective:To establish a glioma cell line U251 stably expressing programmed cell death 4(PDCD4)gene,and to observe the influence of exogenous PDCD4 gene on the proliferation and cell cycle of U251 cells.Methods:Recombinant eukaryotic expression vector pEGFP-PDCD4 was transfected into human glioma cell line U251 by Lipofectamine 2000,and the U251 cells stably expressing PDCD4 were established by G418 selection.Reverse transcription polymerase chain reaction(RT-PCR)and Western blotting were employed to detect the expression of PDCD4 mRNA and protein.Furthermore,cell proliferation and colony forming ability were determined by cell counting and colony formation assay;the cell cycle was detected by FACS.Results:High expression of PDCD4 mRNA and protein was observed in U251 cells transfected with pEGFP-PDCD4,whereas no PDCD4 mRNA and protein expression was detected in the non-transfected and vector-transfected cells.Further,cells transfected with pEGFP-PDCD4 grew more slowly and had lower colony formation rate than cells of the other two control groups(P

The lecture, role play, combination of team learning and self-directed learning, group presentation,these multiple pedagogies were adopted in the teaching of selective medical English course.The questionnaire result showed this multiple teaching strategy can foster the students' interest in medical English, enhance their awareness of active learning and increase the ir learning efficacy, which is an effective teaching model, and can be applied and promoted in the medical English teaching.

Objective To estimate reliability of magnetic resonance elastography (MRE) in measuring liver stiffness of volunteers and patients with chronic liver disease and to assess inter-rater consistency.Methods MRE was performed on a 3.0 T scanner in all subjects,including 24 volunteers (control group) and 64 patients with liver disease (chronic liver disease group).Liver stiffness was measured blindly by two raters.The pathological fibrosis score was applied as a standard reference for liver fibrosis in 22 patients.The intraclass correlation coefficient (ICC) was used to evaluate inter-rater reliability.The differences of liver stiffness between two groups were evaluated using non-parametric MannWhitney U test.Spearman analysis was used to analyze the correlation between fibrosis stages and liver stiffness.Results The intraclass correlation coefficient of liver stiffness was perfect (ICC =0.99,P ＜ 0.01)between two raters.There was significant difference of mean stiffness between control group and patient group (U =90.5,P ＜0.01) with(2.35 ±0.34) kPa and(4.17 ± 0.47) kPa,respectively.The correlation between fibrosis stage (3,3,5,5 and 6 patients in fibrosis stage S0,S1,S2,S3 and S4) and stiffness (2.13,3.25,3.82,5.45 and 7.35 kPa) was very strong (r =0.96,P ＜0.01).Conclusion MRE is a reliable and promising tool to measure liver stiffness and to assess liver fibrosis.

Objective To study the clinical values of polyvinyl alcohol chemoembolization and bronchial arterial infusion(BAI) in interventional therapy of central bronchogenic carcinoma.Methods 45 pathologically proved cases of central bronchogenic carcinoma in mid or late period were randomly divided into two groups(control group in 24 cases and experimental group in 21 cases).After detecting the nourishing arteries of bronchogenic carcinoma with angiography,the treating processes were performed.Control group : anticancer drugs were poured into bronchial artery.Experimental group : the feeding arteries of tumors were co-axially superselective catheterized with microcatheters,the anti-neoplasm agents were poured into everyone of feeding arteries according to the blood supply,then the feeding arteries were embolized by polyvinyl alcohol.Results Control group : the obvious effective rate(CR+PR) was 50%.Experimental group:the obvious effective rate was 90.4%.Two cases in control group obtained the chances of radical operation and chemotherapy were performed after operation,the other 22 cases were undergone synthetic treatment.Seven cases in experimental group obtained the chances of radical operation and chemotherapy were performed after operation,14 cases were undergone synthetic treatment.Three cases in control group died and none of in experimental group died in the follow-up of 2~24 months.Conclusion The short-term effect of polyvinyl alcohol embolization is better than those of BAI for interventional therapy of central bronchogenic cancer,but the long-term effect needs a large number of cases to access the real clinical value.

Objective To study the expression of urokinase-type plasminogen activator (uPA) and its relation with expression of receptor (urokinase-type plasminogen activator receptor uPAR) in epithelial ovari- an cancer (OEC) and with the clinic prognosis.Methods Expression of uPA and uPAR protein was detected by Streptavidin-biotin-HRP in 68 cases of epithelial ovarian cancer and compared with that in 10 cases of borderline tumor,10 cases of benign tumor and 10 cases of normal tissue,and correlation between them was analyzed.The different expression groups of uPA was correlated with the prognosis of ovarian epithelial can- cer.The expression of uPA showed a correlation with short survival time (P

Objective To investigate the effect of euphorbiasteroid on inducing the apoptosis of HL-60 cells and demonstrate whether the Fas/FasL signaling pathway is involved in the induction of apoptosis. Methods HL-60 cells were treated with dose of 2.5,10,40 μg/ml of euphorbiasteroid in vitro for 24 h respectively.After that,cell counting Kit-8 was used to detect cell proliferation.The morphology of HL-60 cells were observed under light and fluorescent microscopy.The early cell apoptosis was detected by using flow cytometry with Annexin Ⅴ-FITC /PI double staining.The expressions of Fas,FasL,caspase-8 and caspase-3 mRNA were analyzed by the method of RT-PCR.The activities of caspase-8 and caspase-3 were examined by chromatometry.Results Compared with 1640 control group,HL-60 cell proliferation was inhibited significant-ly by euphorbiasteroid.The inhibition rates were (34.9 ±3.7)%,(54.6 ±5.2)% and (61.3 ±4.3)%respectively.Moreover,HL-60 cells exhibited typical morphological features.Early cell apoptosis rates of HL-60 cells were (23.4 ±3.1)%,(35.7 ±4.3)% and (53.2 ±3.9)% respectively.Furthermore,the expressions of Fas,FasL,caspase-3 and caspase-8 mRNA were up-regulated significantly after euphorbiasteroid administration in a dose-dependent manner (P<0.01 ).After treated with euphorbiasteroid,the activities of caspase-8 and caspase-3 were significantly enhanced (P<0.01 ).Conclusion The up-regulation effect of euphorbiasteroid on Fas/FasL signaling pathway might contribute to the apoptosis of HL-60 cells.

Objective:To investigate the effectiveness and safety of manual acupuncture for memory loss and sleep quality in chronic insomniacs.Methods:A total of 60 eligible participants were enrolled and randomized into either a treatment group or a control group,with 30 cases in each group.The treatment group was intervened by manual acupuncture whereas the control group was given sham acupuncture.In the two groups,the interventions were offered once every other day and three times a week,for 8 weeks in total.Before and after the treatment,Pittsburgh sleep quality index (PSQI) and eventrelated potentials (ERPs) were used to assess the patients' sleep quality and memory,respectively.Meanwhile,adverse events were monitored and recorded.Results:After 8-week treatment,both the treatment group and the control group showed a significant decrease in the PSQI global score (P＜0.001,P＜0.01),and the decrease in the treatment group was more significant than that in the control group (P＜0.001).The intra-group comparisons of ERPs indicated that,the latencies of N1 and P3 were shortened and the amplitudes of N1 and P3 were increased in the treatment group after the intervention,and the differences were statistically significant (P＜0.05,P＜0.001);in the control group,there were no significant changes in the latency and amplitude after the treatment (P＞0.05).The between-group comparisons of ERPs showed that the treatment group was more effective than the control group in shortening the latency of P3 (P＜0.01).Conclusion:Acupuncture can be a safe and effective treatment option for chronic insomnia coupled with memory impairment.

BACKGROUND: A large amount of in vivo and in vitro experiments have confirmed that, basic fibroblast growth factor (bFGF) has been widely utilized in various tissues and cells, it can facilitate the wound healing.OBJECTIVE: To observe the efficacy and feasibility of gene gun-mediate delivery of human bFGF on the healing of deep partial thickness bum wounds.DESIGN, TIME AND SETTING: Randomized design,an observational trial was performed at the Military Central Laboratory of Changhai Hospital in the Second Military Medical University of Chinese PLA between December 2007 and October 2008.MATERIALS: SD rats of clean grade, weighing 200-250 g, irrespective of genders, ware involved in this study.METHODS: Natural human bFGF gene was recombined and optimized, then eukaryotic expression vector pCI-neo-bFGF was constructed taking pCI-neo as a vector, and transfeoted with human embryonic kidney cells 293 T cells. Dot blot and Western blot methods were utilized to determine the bFGF expression. Rat model of deep partial thickness burn wounds was processed into transgene process using gene gun technique, pCI-neo-bFGF-transfected ones served as experiment group while pCI-neo-transfected ones served as controls.MAIN OUTCOME MEASURES:Wound healing time was recorded and the efficacy was evaluated. The contents of hydroxyproline and collaganase Ⅰ in burn wound tissues were determined at 24 hours, 48 hours, 96 hours, 7 days, 10 days and 14 days following transgene process.RESULTS: the recombinant pCI-neo-bFGF was transfected with human embryonic kidney 293T cells. Dot blot and Western blot analysis have showed that, the constructed pCI-neo-bFGF expression vector could express human bFGF, and the expression of synthesized gene was remarkably higher than that of natural gene under fluorescence microscope; gene gun-mediated transgene experiment have showed that, the wound healing time was (13.00+1.31) days in the experiment group and (14.75±1.28) days in the control group, with significant differences (P＜0.05). The contents of hydroxyproline and collagenase Ⅰ reached a peak at 5 days after the injury, that is 48 hours after transfection, and then gradually decreased and maintained at a certain level. The experiment group had higher hydroxyproline levels compared with control group at different time points (P＜0.05, P＜0.01); the collagenase Ⅰ in the experiment group was notably higher than that in the control group at 48 hours and 96 hours after transfection (P＜0.01).CONCLUSION: Gene gun-mediated delivery of human bFGF can short the time of wound healing, increase the contents of hydroxyproline and collagenase Ⅰ during the healing period, accelerate the healing of deep partial thickness burn wounds.

Objective To observe the apeptosis in liver injury following limbs ischemia-reperfusion(IR) in rats and the protective effects of taurine.Methods The model of limbs ischemia-reperfusion injury was established.30 Wistar rats were randomized into 3 groups: control group,IR group and tanrine + reperfusion group (TR group) (n = 10 for each group).The levels of malondialdehyde (MDA) and xanthineoxidas (XOD), calcium and myeloper-oxidase (MPO) in the liver tissue were measured.DNA fragmentation was observed and analyzed by agarose gel e-lectrophoresis.Apoptosis was detected by TUNEL methods.The morphologic changes were observed with HE stai-ning.Results Compared with control group,the values of MDA, XOD, MPO, calcium in liver tissue were increased significantly in IR group (P<0.01), but the values of those in TR group were lower than in IR group (P<0.01).The percentage of apeptosis cell was higher in IR group than in control group(P<0.01).Compared with IR group, the percentage of apoptosis cell was lower in TR group (P<0.01).IR group presented DNA ladder pattern, while TR group showed no specific DNA ladder pattern in agarose gel electrophoresis.Conclusion Apoptosis participates in the liver injury after limb ischemia-reperfusion.Taurine can mitigate the liver injury and apoptosis after limb is-chemia-reperfusion injury in rats.