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OBJECTIVE:To optimize the ethanol extraction process of anti-atherosclerosis effective fraction of Lindley eupatorium. METHODS:Using the contents of hyperin,quercetin,kaempferol and jaceosidin as indexes,the effects of ethanol volume fraction,liquid-solid ratio,extraction time and extraction times on ethanol extraction technology were investigated. By multiple index scoring method,orthogonal test was designed to optimize extraction technology,and validation test was conducted. RESULTS:The optimal extraction technology was 70% ethanol as extraction solvent with solid-liquid ratio of 12 : 1,reflux extracting for 3 times,2 h each time. Results of 3 batches of validation tests showed that the contents of 4 marker components were in high level,being 5.305 3,1.560 1,1.986 5,6.703 6 mg/g in average(RSD<1.1%,n=3). Results of validation tests were stable and reliable,and were in good agreement with the results of orthogonal tests. CONCLUSIONS:The optimized ethanol reflux extraction technology of L. eupatorium is stable and reliable,and performs well. Energy consumption and cost are suitable for industrial production.The results provide technology basis for further purification and enrichment of anti-atherosclerosis effective components of L.eupatorium.
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Objective To investigate the molecular mechanisms of PLCεin regulating the invasion and migration of human bladder cancer cells in vitro.Methods After cells treated with recombinant adenovirus , the migratory/in-vasive abilities of T24 cells were explored by wound healing and Transwell chamber cell migration and invasion as -say;RT-PCR was used to detect the mRNA levels of PLCε;The protein levels of PLCε,PKCα,PKCβ, TBX3 and E-cadherin were determined by Western blot;QRT-PCR was used to detect the mRNA levels of TBX3 and E-cad-herin.Results It was confirmed by digesting and sequencing that the recombinant adenovirus had been constructed successfully .The expression of PLCε mRNA and PLCε protein were both decreased after the infection of Ad-shPLCε.Wound healing and Transwell chamber cell migration/invasion assay showed that Ad-shPLCε treatment could inhibit the migratory and invasive activity of bladder cancer cells(P<0.05).The results of Western blot indicated that the expression of PKCα/βin membrane decreased ( P<0.05 ) , and phosphorylation level of PKCαand PKCβwas reduced .QRT-PCR and Western blot analysis demonstrated that the expression level of TBX 3 de-creased , but the expression level of E-cadherin increased .Conclusions PLCε shRNA can inhibit migratory and invasive ability of bladder cancer cells through PKCα/β/TBX3/E-cadherin pathway .
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This study was aimed to summarize main-bone structure of cantharidin-based small molecules and facts about three typical candidates including its origin, physical-chemical properties and general synthetic approaches. Basic chemical and pharmacological information as well as development of anti-cancer activities, which were related to cantharidin, norcantharidin and their analogues were reviewed, especially the relationship between the structures and their inhibitory activity and selectivity of serine-threonine protein phosphatases PP1, PP2A and PP2B in cancer treatment. The designs and developments of new biological cantharidin-based small molecules were also reviewed.
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Extraction process plays an important role in Chinese herbal medicine applications. Traditional extraction method has some disadvantages, such as long processing time, high solvent consumption and low efficiency. Thus, some new technologies and methods have been introduced in this paper, including ultrasonic extraction, microwave extraction, supercritical fluid extraction. On the basis of advantages and disadvantages of traditional Soxhlet extrac-tion, this paper mainly described the principles, characteristics, influencing parameters and applications.