The high production inulase strain was screened from the soil sample where burdock planted in Qin Village,Bayou Town,Pei County,Xuzhou.Inulase activity were determined which produced by 40 strains separated from soil.Three mold stains,C122803、D081506 and D081513,which had higher ability of producing inulase were obtained by using transparent circle method as initial screening and rocker method as re-screening.Enzyme activity of the three strains were 1.411U/ml,1.895U/ml,1.792U/ml,separately.Enzyme activity of D081506,1.895U/ml,was the highest.The fermentation conditions of D081506 were studied and the optimized conditions were lappa juice 2.0%,yeast extraction 1.6%,(NH4)2SO4 0.5%,NaCl 0.5%,K2HPO4 0.5% and pH 5.0.Inulase activity of D081506 was 2.9578U/ml which increased 56.09% under the condition of 27℃,140r/min,24h.

OBJECTIVETo study the apoptotic effect on the squamous cell carcinoma cell line TCa83 induced by recombined adenovirus vector containing TRAIL gene and CMV promoter.

METHODSThe TCa83 cell line was firstly infected with different titre of AdCMV-EGFP containing enhanced green fluorescence protein gene (EGFP) as control, and investigated the transducing rate through fluorescence to obtain the definite titre. Then TCa83 cell line was infected with AdCMV-TRAIL in proper titre, and TRAIL gene was detected by means of RT-PCR. After TCa83 cell line was infected with AdCMV-TRAIL and AdCMV-EGFP at day 1, 3, 5, 7, the activity of TCa83 cell line were evaluated by MIT and the apoptosis were detected by flow cytometer.

RESULTSProper titre was of 1,000 particles/cell, and TCa83 cell line could be infected 100% in this titre. TRAIL gene was detected by RT-PCR after infected with AdCMV-TRAIL. The activity of TCa83 decreased in both groups, but the AdCMV-TRAIL group decreased more sharply than AdCMV-EGFP group (P < 0.001). Both AdCMV-TRAIL and AdCMV-EGFP could lead to apoptosis of TCa83 cells, but the AdCMV-TRAIL, function stronger than AdCMV-EGFP. Especially there was remarkable statistic difference between two groups (P < 0.0001).

CONCLUSIONAdCMV-TRAIL could effectively decrease the activity of TCa83 cell line and induce apoptosis.

Adenoviridae ; Apoptosis ; Carcinoma, Squamous Cell ; Cell Line ; Genetic Vectors ; Green Fluorescent Proteins ; Humans ; Promoter Regions, Genetic

As a natural product with a long history of medicinal use, parthenolide has aroused great interest of chemists and biologists. Existing studies have shown that it has anti-inflammatory, antitumor and other pharmacological activities, and also revealed its action on NF-κB signaling pathway, DNMT1 enzyme and Wnt/β-catenin signaling pathway. But its biological targets remain to be elucidated systematically. Proteolysis Targeting Chimeras (PROTAC) provides a new strategy for target discovery of natural products, which can be used to explore the panorama of protein changes in cells through proteomic investigation, so as to analyze their potential targets. Based on this idea, current study designed and synthesized 20 parthenolide-derived degraders. After measured their antitumor activity in vitro, selected compounds were carried out the proteomic experiment. Finally, 139 down-regulated differentially expressed proteins were identified and the discovery of parthenolide interacting protein was preliminarily explored.