Based on previous report that the Chinese herb Ligustrum lucidum (LL) extract directly inhibited hepatitis C virus (HCV) replicase (NS5B) activity, the active components of LL extract to inhibit HCV NS5B activity and their inhibition mode were investigated in this study. LL extract was separated using ethyl acetate and thin layer chromatography (TLC). The inhibitory activity of separated fractions on HCV NS5B was analyzed by the inhibitory assay of NS5B activity. The results showed that only fractions 1 and 2 inhibited NS5B activity, and fraction 2 possessed higher inhibitory activity than fraction 1. HPLC analysis combined with inhibitory assays indicated that ursolic acid and oleanolic acid are the active components within fractions 1 and 2 to inhibit NS5B activity, separately. Moreover, oleanolic acid possessed higher inhibitory activity than ursolic acid. Further inhibition mode analysis found that both oleanolic acid and ursolic acid suppressed NS5B activity as noncompetitive inhibitors. The Ki values of ursolic acid and oleanolic acid were about 4.7 microg x mL(-1) (10 micromol x kg(-1)) and 2.5 microg x mL(-1) (5.5 micromol x kg(-1)), respectively. Taken together, these results demonstrated that oleanolic acid and ursolic acid suppressed NS5B activity as noncompetitive inhibitors, implying that the two natural products have potential value for HCV therapy.

Objective To investigate the feasibility and safety of surgical device closure of doubly committed sub-arterial ventricular septal defect via left sub-axillary.Methods A total of 45 patients diagnosed as doubly committed sub-arterial ventricular septal defect (dcVSD) with transthoracic echocardiography (TTE) and transesophageal echocardiography(TEE) were enrolled from June 2014 to August 2016 in Henan Children Heart Center,Henan Provincial People's Hospital.There were 39 males and 6 females,with the mean age of (2.2 ±2.1) years old(0.5-8.0 years),the body weight (13.8 ± 7.1) kg(7.0-34.1 kg),the defect size (4.5 ± 1.0) mm (3.0-8.0 mm).After general anesthesia,the patients were in supine and evaluated by TEE which indicated whether they were fit to closure.Then,they were turned to the right lateral position while this technique was determined.A vertical incision of 2-3 cm was made between the third and the fifth intercostal space and invasion in thoracic space via fourth intercostal space.Puncture was done at the anterior surface of right ventricular outlet tract to build a delivery tract.The occluder was released and the VSD was occluded under transesophageal echocardiography guidance.Results Forty-one patients had a successful surgical dcVSD closure with asymmetric occluders sized (6.0 ± 1.5) mm(4-10 mm).Among 4 failure cases,2 cases (4.4％) were switched to open-heart surgical repair,1 case (2.2％) due to device related aortic regurgitation,the rest 1 case (2.2％) experienced a dislocation of occluder into pulmonary artery and was converted to surgical repair after retrieve of occluder.Trivial residual shunt was detected in 2 cases (4.4％) postoperatively,a spontaneous closure was observed by 1 month follow-up and 3 months follow-ups,respectively.All the patients were discharged 5 to 8 days after the operation.With a follow-up of (10.4 ±5.0) months [3-24 months],there were no complications such as pericardial effusion,displacement of device,atrioventricular block or new valvular dysfunction.Conclusions Minimally invasive device closure of doubly committed sub-arterial ventricular septal defect via left sub-axillary is a feasible and safe treatment for closure of dcVSD.This technique has advantages of minor wound,less exudation,covert incision,however,long term follow-up is necessary.

Objective To investigate the effect ofmiR-184 on proliferation of neural stem cells (NSCs) and its mechanisms in mice.Methods The pHBLV-U6-GFP-miR-184 over-expression plasmid and pHBLV-U6-GFP-miR-184 inhibitor plasmid were used to construct recombinant lentivirus.And the NSCs derived fiom subventricular zone of E14d CD1 mouse were confirmed by immunofluorescence assay.There were four groups that contain a miR-184 overexpression group,a miR-184 inhibitor group and two control groups.The NSCs which infected with lentiviral vectors were selected for puromycin resistance for 5-7 days,and then surviving cells were cultivated to three generations.The expression level ofmiR-184 was detected by real time-quantitative PCR (RT-qPCR).And the target genes ofmiR-184 were predicted through TargetScan,IRTarBase and MiRanda,and were confirmed by Western blotting and RT-qPCR.The cells in the four groups were culttared under proliferating conditions incorporated bromodeoxyuridine (BrdU) in cell proliferation analyses.The protein expressions of Hesl and Hes5,the target proteins of Notch signaling pathways,and their mRNA expressions were detected by Western blotting and RT-qPCR.Results There were 90％ of cells in each group expressing both Nestin and Sox2.The miR-184 level in the miR-184 overexpression group was 67.63±7.53 times of that of the control group,with significant difference (P＜0.05).The percent of BrdU+/DAPI+ of the miR-184 overexpression group was 1.47±0.05 times of that in the control group,with significant difference (P＜0.05);and the percent of BrdU+/DAPI+ of the miR-184 inhibitor group was 0.84±0.03 times of that in the inhibitor control group,with significant difference (P＜0.05).Numbl was a target gene ofmiR-184 indicated by IRTarBase and MiRanda.The miR-184 could inhibit Numbl protein expression;the Numbl protein expression level in the miR-184 overexpression group was 0.73±0.07times of that in the control group,and the Numbl protein expression level in the miR-184 inhibitor group was 1.30±0.05 times of that in the control group,with significant difference (P＜0.05);but miR-184 did not change the Numbl mRNA level.The miR-184 could activate Notch signaling pathway through inhibiting the Numbl protein expression,and increase the Hes1 and Hes5 protein and mRNA expression levels (P＜0.05).Conclusion The miR-184 promotes the NSCs proliferation through inhibiting the Numbl protein translation and further activating the Notch signaling pathway.

Objective To investigate the effect of galangin on proliferation and apoptosis of glioma cells in vitro.Methods (1) The glioma cells U87 and U25 1were divided into blank control group,DMSO group,100,200,300 and 400 μmol/L galangin treatment groups.MTT was used to study the effects of drugs on the proliferation of U251 and U87 cells.(2) Hoechest staining was used to observe cell apoptosis in the presence of different concentrations of galangin (0,100 and 200 μmol/L).(3) Flow cytometry was employed to detect the apoptosis of U251 and U87 cells in the presence of different concentrations of galangin (1 00 and 200 μmol/L).(4) Western blotting was employed to detect the expressions of apoptosis-related protein 3-Catenin,B-cell lymphoma-2 (Bcl-2),Bcl-2 related protein gene (Bax),cleaved-caspase-3,cleaved-caspase-9 and poly (ADP-ribose) polymerase (PARP) in the presence of different concentrations of galangin.Results (1) The proliferation of U251 and U87 cells was obviously inhibited atter 100,200,300 and 400 μmol/L galangin treatments,and dose-effect relation was noted.The concentrations of galangin at half rate of inhibition (IC50) were 281,321,276 and 229 μmol/L in U251 cells,and 289.4,261.1,247.4 and 225.3 μ mol/L in the U87 cells after 100,200,300 and 400μmol/L galangin treatments for 24 h.(2) Under the action of galangin,corresponding increase in apoptosis rates of U251 and U87 cells was noted following the increase of galangin concentrations (0,100 and 200 μmol/L),with significant differences (P＜0.05).(3) The detection of cell apoptosis by flow cytometry found similar changes.(4) Western blotting results indicated that galangin at the concentration of 0,100 and 200 μmol/L could significantly decrease the expressions of apoptosis-related protein 3-Catenin and Bcl-2,and increase the Bax,cleaved-caspase-3 and cleaved-caspase-9,and cleaved-PARP expressions;significant differences were noted between each two concentrations (P＜0.05).Conclusion Galangin can inhibit proliferation of glioma cells U251 and U87,and induce mitochondrial pathway of apoptosis via Wnt/β-Catenin signaling.

Cancer is one of the most important diseases threatening human health. Frequently-used traditional cancer treatment methods, like radiotherapy, chemotherapy and surgery, have serious toxic side effects and limitations. The widely-used drug delivery carriers (liposomes, nanoparticles, etc.) have also possessed many issues such as drug leakage and incomplete loading in the late clinical stage. Currently, using tumor-targeting vectors to deliver anti-tumor drugs or small molecules is one of the promising strategies for mediating safe and effective tumor therapy. In recent years, bacterial-derived non-replicating minicells, which are nanoscale non-nucleated cells produced during abnormal bacterial division, have got more and more attention. With a diameter of 200-400 nm, minicells have a large drug loading capacity. Meanwhile, the surface of minicells are able to be modified to load the assembly of antibodies/ligands that bind to tumor cell surface specific antigens or receptors, which can significantly improve tumor targeting of minicells. This tumor-targeting nanomaterials of minicells not only are used to deliver anti-tumor chemotherapeutic drugs, functional nucleic acids or plasmids encoding functional small molecules to mammalian cells, but also greatly increase drug loading and reduce drug penetration. Thus, the use of minicells combining with chemical therapy could help reduce the toxicity and maximize the effectiveness of the drug in the body. This paper summarizes the research and development of production purification, drug loading, tumor cells targeting, and internalization process of minicells, as well as its use in the delivery of anti-tumor drugs, to provide some information for the development and utilization of minicell carriers.
Animals ; Drug Carriers ; Drug Delivery Systems ; Humans ; Nanoparticles ; Neoplasms ; Plasmids