In vivo 3D fluorescent image remains a technological barrier for biologists and clinical scientists although green fluorescent protein(GFP)imaging has long been performed rather well at cellular level.Meanwhile,robust...In vivo 3D fluorescent image remains a technological barrier for biologists and clinical scientists although green fluorescent protein(GFP)imaging has long been performed rather well at cellular level.Meanwhile,robust enough portable devices are also challenging lab-on-a-chip advocators who wish their designs to be nurtured by the end users.This work is dedicated to propose a conceptually innovated transparent soft PDMS avian eggshell to directly tackle the above two goals.Here,an"egg-on-a-chip"scheme is originally developed and demonstrated by a newly developed PDMS"soft"process method.Unlike its ancestor–the conventional"lab-on-a-chip"(LOC)which is basically chemically based,the current"egg-on-a-chip",intrinsically inherited with biological natures,opens a way to integrate biological parts or whole system in a miniature sized device.Such biomimics system contains much condensed environmental evolutional tensor inside than those of the existing LOC compacted with artificial components which however are quite difficult to incorporate various life factors inside.Owning unique advantages,a series of transparent PDMS whole"eggshells"have been fabricated and applied to culture avian embryos up to 17.5 days and chimeric eggshells were engineered on normal eggs.In addition,X-stage embryos were successfully initiated in such system and pre-chorioallantoic membrane was observed.Further,limitation of the present process was interpreted and potential approach to improve it was suggested.With both high optical transparency and engineering subtlety fully integrated together,the present method not only provides an ideal transparent imaging platform for studying functional embryo development including life mystery,but also promises a future strategy for"lab-on-an-egg"technology which may be important in a wide variety of either fundamental or practical areas.展开更多
In order to develop a model for screening the agonists of human β2-adrenoceptor from Chinese medicinal herbs extracts, we used a cell-based functional assay based on a common G protein-coupled receptor （GPCR） regul...In order to develop a model for screening the agonists of human β2-adrenoceptor from Chinese medicinal herbs extracts, we used a cell-based functional assay based on a common G protein-coupled receptor （GPCR） regulation mechanism and destabilized enhanced green fluorescent protein （d2EGFP） reporter gene technique. The positive cell clone was confirmed by real-time polymerase chain reaction （PCR） and imaging analysis. To assess the value of this model, we screened over 2000 high performance liquid chromatography （HPLC）-fractionated samples from the ethanol extracts of Chinese medicinal herbs. Six fractions （isolated from Panax japonicus, Veratrum nigrum, Phellodendron amurense, Fructus Aurantii lmmaturus, Chaenomeles speciosa, and Dictamnus dasycarpus） showed significant effects on active reporter gene expression, three of which （isolated from Phellodendron amurense, Fructus Aurantii lmmaturus, and Chaenomeles speciosa） were selected for further concentration response analysis and the half maximal effective concentration （EC1/2 max） values were 4.2, 2.7, and 4.8 μg/ml, respectively. Therefore, this reporter gene assay was suitable for screening β2-adrenoceptor agonists. The results suggest that the six herbal extracts are the possible agonists of β2-adrenoceptor.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.51376102)
文摘In vivo 3D fluorescent image remains a technological barrier for biologists and clinical scientists although green fluorescent protein(GFP)imaging has long been performed rather well at cellular level.Meanwhile,robust enough portable devices are also challenging lab-on-a-chip advocators who wish their designs to be nurtured by the end users.This work is dedicated to propose a conceptually innovated transparent soft PDMS avian eggshell to directly tackle the above two goals.Here,an"egg-on-a-chip"scheme is originally developed and demonstrated by a newly developed PDMS"soft"process method.Unlike its ancestor–the conventional"lab-on-a-chip"(LOC)which is basically chemically based,the current"egg-on-a-chip",intrinsically inherited with biological natures,opens a way to integrate biological parts or whole system in a miniature sized device.Such biomimics system contains much condensed environmental evolutional tensor inside than those of the existing LOC compacted with artificial components which however are quite difficult to incorporate various life factors inside.Owning unique advantages,a series of transparent PDMS whole"eggshells"have been fabricated and applied to culture avian embryos up to 17.5 days and chimeric eggshells were engineered on normal eggs.In addition,X-stage embryos were successfully initiated in such system and pre-chorioallantoic membrane was observed.Further,limitation of the present process was interpreted and potential approach to improve it was suggested.With both high optical transparency and engineering subtlety fully integrated together,the present method not only provides an ideal transparent imaging platform for studying functional embryo development including life mystery,but also promises a future strategy for"lab-on-an-egg"technology which may be important in a wide variety of either fundamental or practical areas.
基金Project (No. 30873103) supported by the National Natural Science Foundation of China
文摘In order to develop a model for screening the agonists of human β2-adrenoceptor from Chinese medicinal herbs extracts, we used a cell-based functional assay based on a common G protein-coupled receptor （GPCR） regulation mechanism and destabilized enhanced green fluorescent protein （d2EGFP） reporter gene technique. The positive cell clone was confirmed by real-time polymerase chain reaction （PCR） and imaging analysis. To assess the value of this model, we screened over 2000 high performance liquid chromatography （HPLC）-fractionated samples from the ethanol extracts of Chinese medicinal herbs. Six fractions （isolated from Panax japonicus, Veratrum nigrum, Phellodendron amurense, Fructus Aurantii lmmaturus, Chaenomeles speciosa, and Dictamnus dasycarpus） showed significant effects on active reporter gene expression, three of which （isolated from Phellodendron amurense, Fructus Aurantii lmmaturus, and Chaenomeles speciosa） were selected for further concentration response analysis and the half maximal effective concentration （EC1/2 max） values were 4.2, 2.7, and 4.8 μg/ml, respectively. Therefore, this reporter gene assay was suitable for screening β2-adrenoceptor agonists. The results suggest that the six herbal extracts are the possible agonists of β2-adrenoceptor.
