Aim To construct a screen assay of ?-secretase inhibitors.Methods A human ?-secretase cDNA was cloned by RT-PCR and then reconstructed in an expression vector.The recombinant ?-secretase fusion protein was expressed and purified from Escherichia coli.A screen assay of ?-secretase inhibitors was constructed in vitro using a recombinant A ?-fusion protein as substrate.The enzyme reaction was detected by SDS-PAGE and an ELISA method using metal chelating plates to catch the substrate.Results A human ?-secretaae cDNA was cloned and expressed in escherichia coli.The ?-secretase enzyme reaction can be detected by two kinds of method: SDS-PAGE and ELISA.The IC_(50) for an inhibitor statVal determined by this method was in good agreement with those reported in the literature.Seventy seven species of Chinese traditional medical plants were screened using this method and seventeen species show weak inhibitory activity.Conclusion A screen assay of ?-secretase inhibitors was successfully constructed and can be used to screen natural products.

Three triterpenoid saponins were isolated from 90％ ethanolic extract of the seeds of Aes-culus turbinata Bl.. Their structures were identified on the basis of spectral data as escin IVc:22α-O-tigloyl-28-O-acetylprotoaescigenin 33-O-[β-D-glucopyranosyl (1→2)][β-D-glucopyranosyl (1→4)]-β-D-glucopyranosiduronic acid (Ⅰ), isoescin Ia: 21β-O-tigloyl-28-O-acetylprotoaescigenin-3β-O-[β-D-glucopy-ranosyl (1→2)][β-D-glucopyranosyl (1→4)]-β-D-glucopyranosiduronic acid (Ⅳ), and isoescin Ib:21β-O-angeloyl-28-O-acetylprotoaescigenin-3β-O-[β-D-glucopyranosyl (1→2)][β-D-glucopyr anosyl (1→4)]-β-D-glucopyranosiduronic acid (Ⅴ). Escin IVc and isoescins Ia, Ib were obtained for the first time from thisplant. Escins Ia, Ib, IVc and isoescins Ia, Ib had inhibitory effect against recombinant HIV-1 protease atconcentration of 100 μmol/L in vitro.

To explore the effects of Quyu Xiaoban Capsule (QXC) on blood lipid level and platelet activity in coronary heart disease (CHD).A randomized single blinded trial was adopted in 62 cases of CHD. Thirty one cases (control group) were treated with routine western medicine and the rest (QXC group) with QXC added. Serum levels of total cholesterols (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL C) and platelet-activating ? granule membrane protein (GMP 140) were detected before and after treatment.After ten weeks of treatment, GMP 140, TC and TG levels were decreased to various degrees in QXC group, the differences being significant as compared with those in control group and those in QXC group before treatment (P0 05).[Conclusion]QXC can decrease the blood lipid level and improve platelet activity and has a certain effect in preventing and treating CHD.

Objective To evaluate the short-term effectiveness of Amplatzer device closure for congenital heart diseases.Methods Under X-ray fluoroscopy or/and transthoracic echocardiography(TTE),percutaneous puncture of the femoral artery or vein was conducted and the Amplatzer occluder was passed through the catheter.Effectiveness evaluation by transthoracic echocardiography,electrocardiography(ECG),and X-ray examination was applied at 24 hours,1 month,6 months,and every 1 year after the procedure.Results In 1 case of perimembranous ventricular septal defect(VSD),the detachment of the Amplatzer occluder to the arch of aorta was due to an inadequate small size.This patient was immediately operated with successful removal of the device through the femoral artery and later underwent a re-deployment of a larger-sized Amplatzer occluder closure successfully.The remaining 29 patients with atrial septal defect(ASD),patent ductus arteriosus(PDA),or perimembranous ventricular septal defect underwent a successful deployment of the Amplatzer occluder on one session without complications.The operation time was 20~90 min (38?16 min),the X-ray exposure time was 5~45 min(18?10 min),and the length of hospital stay was 3~7 days(4?2 days).Conclusions Amplatzer occluder transcatheter closure of congenital heart diseases has advantages of simplicity of deployment,good safety,and high success rate.This procedure is suitable for patients with secundum ASD,membranous VSD,and PDA.

0.05). Conclusion:The effect of intravenous vecuronium made in Xianju Pharmaceutical Co. for neuromuscular blockade is definite and safe.

Objective To review the study of noninvasive imaging methods for evaluating liver fibrosis.Meth- ods The current literatures on the use of the ultrasonography,CT and MRI for the evaluation of liver fibrosis were reviewed.The principles,applications and advancement of each imaging methods were described and summarized re- spectively.The features of the newly developed imaging techniques were also discussed.Results In addition to the morphologic information,the imaging examinations can also provide functional information about the circulation sta- tus,diffusion and metabolism features of liver.The potential diagnostic value of MR elastography for liver fibrosis has been addressed.Conclusion The imaging examinations,especially the functional MRI techniques,are reliable noninvasive alternatives for the evaluation of hepatic fibrosis,with bright potentiality for clinical application.

Objective:To study the preparation technique and quality standard for new Shengfa pills. Methods: The best water extraction process of the medicinal herbs was optimized by orthogonal test using stilbene glucoside content and dry extract percent as the indices. The content of stilbene glucoside was determined by HPLC, and Hyssop and Chinese angelica in the preparation were identified by TLC. Results:The optimized preparation conditions were as follows:12-fold water was used in the 3-time extraction with 1 h for each time. The spots in TLC were clear without interference from the negative control. Conclusion:The preparation process is reasona-ble and feasible in techniques with controllable quality.

Objective To investigate the effect of bortezomib combined with doxorubicin on lymphoblastic lymphoma cell line Molt-4.Methods Molt-4 cells were cultured in the presence of bortezomib and doxorubicin,cell viability was monitored by CCK-8 and trypan-blue exclusion.Apoptosis was detected by flow cytometry and mitochondrial membrane potential,expression of Fas was also measured with flow cytometry.Results Molt-4 cell proliferation was substantially inhibited in concentration-dependent manners when treated with either bortezomib or doxorubicin.The combination of both drugs synergistically inhibited Molt-4 cell proliferation at 48 hours [(57.24±0.10) ％].Combination therapy further enhanced bortezomib and doxorubicin induced apoptosis [48 h (23.08±1.25) ％] (P ＜ 0.05).Detection of mitochondrial membrane potential showed that combination therapy could promote apoptosis (15.84 ％,5.38 ％,5.52 ％) but did not significantly change the level of Fas expression (P ＞ 0.05).Conclusion Combination therapy of bortezomib with doxorubicin efficiently inhibits proliferation and induces apoptosis of Molt-4 cells.Activation of mitochondrial and intrinsic apoptotic pathway may play important roles.

To construct a lentiviral shRNA vector targeting human protein phosphatase 1D magnesium-dependent (PPM1D) gene and detect its effectiveness of gene silencing in human gliomas, specific siRNA targets with short hairpin frame were designed and synthesized. DNA oligo was cloned into the pFU-GW-iRNA lentiviral expression vector, and then PCR and sequencing analyses were conducted to verify the constructs. After the verified plasmids were transfected into 293T cells, the lentivirus was produced and the titer of virus was determined. Real-time quantitative PCR and Western blot were performed to detect the PPM1D expression level in the infected glioma cells. PCR and Western blot analyses revealed the optimal interfering target, and the virus with a titer of 6×10(8) TU/mL was successfully packaged. The PPM1D expression in human glioma cells was knocked down at both mRNA and protein levels by virus infection. The expression of PPM1D mRNA and protein was decreased by 76.3% and 87.0% respectively as compared with control group. The multiple functions of human glioma cells after PPM1D RNA interference were detected by flow cytometry and cell counting kit-8 (CCK-8). Efficient down-regulation of PPM1D resulted in significantly increased cell apoptosis and reduced cell proliferation and invasion potential in U87-MG cells. We have successfully constructed the lentiviral shRNA expression vector capable of stable PPM1D gene silencing at both mRNA and protein levels in glioma cells. And our data gave evidence that the reduced cell growth observed after PPM1D silencing in glioma cells was at least partly due to increased apoptotic cell death.

Objective To investigate the role of Twist and vascular endothelial growth factor (VEGF)in the development of epithelial ovarian cancer and their relationship with clinical pathological features.Methods The expressions of Twist and VEGF were detected by immunohistochemical staining (SP)method in 80 cases of epithelial ovarian cancer and 10 paired cases of normal ovarian tissue.Results In the 80 cases of ovarian cancer tissues,the positive expression rate of Twist and VEGF in epithelial ovarian cancer was 85% and 86.25%,respectively,which were remarkably higher than those in normal ovarian tissues (P <0.01).The expressions of Twist and VEGF were closely associated with clinical stages,pathological grades and lymph node metastasis,but not correlated with age or pathological patterns (P >0.05).There was a significant correlation between the expressions of Twist and VEGF in epithelial ovarian cancer (r =0.646,P <0.01).Conclusion The high expressions of Twist and VEGF may play a potential role in the occurrence,development and invasion process of epithelial ovarian cancer.