AIM:To find a rapid and efficient analysis method of gastrointestinal microflora in Pi-deficient(spleen-deficient) rats and to evaluate traditional Chinese drugs.METHODS:Enterobacterial repetitive intergenic consensus...AIM:To find a rapid and efficient analysis method of gastrointestinal microflora in Pi-deficient(spleen-deficient) rats and to evaluate traditional Chinese drugs.METHODS:Enterobacterial repetitive intergenic consensus-PCR(ERIC-PCR) based assay was performed to examine changes of intestinal microflora in two Pi-deficienct animal models and to evaluate the efficacy of four traditional Chinese drugs as well as a probiotic recipe and another therapy in Pi-deficient rats.RESULTS:A molecular marker was identified for Pi-deficiency in rats.The pharmacodynamic evaluation system,including identified molecular markers(net integral area and abundance of DNA bands),Shannon's index for diversity of intestinal microflora,and Sorenson's pairwise similarity coefficient,was established.The four major clinical recipes of traditional Chinese drugs for Pi-deficiency in rats,especially at their medium dose(equivalence to the clinical dose),produced more pronounced recovery activities in Pi-deficient rats,while higher doses of these recipes did not show a better therapeutic effect but some toxic effects such as perturbation deterioration of intestinal microflora.CONCLUSION:Both fingerprint analysis and identified marker can show Pi-deficiency in rats and its difference after treatment.The identified molecular marker may be applied in screening for the active compounds both in relative traditional Chinese drugs and in pharmacodynamic study of Pi-deficiency in rats.展开更多
Chlorpyrifos is a widely used insecticide in recent years,and it will produce adverse effects on soil when applied on crops or mixed with soil.In this study,nested polymerase chain reaction（PCR） and denaturing gradi...Chlorpyrifos is a widely used insecticide in recent years,and it will produce adverse effects on soil when applied on crops or mixed with soil.In this study,nested polymerase chain reaction（PCR） and denaturing gradient gel electrophoresis（DGGE） were combined to explore the bacterial and fungal community successions in soil treated with 5 and 20 mg/kg of chlorpyrifos.Furthermore,isolates capable of efficiently decomposing chlorpyrifos were molecular-typed using enterobacterial repetitive intergenic consensus-PCR（ERIC-PCR）.Under the experimental conditions,degradation of chlorpyrifos in soil was interpreted with the first-order kinetics,and the half-lives of chlorpyrifos at 5 and 20 mg/kg doses were calculated to be 8.25 and 8.29 d,respectively.DGGE fingerprint and principal component analysis（PCA） indicated that the composition of the fungal community was obviously changed with the chlorpyrifos treatment,and that samples of chlorpyrifos treatment were significantly separated from those of the control from the beginning to the end.While for the bacterial community,chlorpyrifos-treated soil samples were apparently different in the first 30 d and recovered to a similar level of the control up until 60 d,and the distance in the PCA between the chlorpyrifos-treated samples and the control was getting shorter through time and was finally clustered into one group.Together,our results demonstrated that the application of chlorpyrifos could affect the fungal community structure in a quick and lasting way,while only affecting the bacterial community in a temporary way.Finally,nine typical ERIC types of chlorpyrifos-degrading isolates were screened.展开更多
基金Supported by The National Natural Science Foundation of China,No.90209059Supported by The National Natural Science Foundation of China,No.90409018
文摘AIM:To find a rapid and efficient analysis method of gastrointestinal microflora in Pi-deficient(spleen-deficient) rats and to evaluate traditional Chinese drugs.METHODS:Enterobacterial repetitive intergenic consensus-PCR(ERIC-PCR) based assay was performed to examine changes of intestinal microflora in two Pi-deficienct animal models and to evaluate the efficacy of four traditional Chinese drugs as well as a probiotic recipe and another therapy in Pi-deficient rats.RESULTS:A molecular marker was identified for Pi-deficiency in rats.The pharmacodynamic evaluation system,including identified molecular markers(net integral area and abundance of DNA bands),Shannon's index for diversity of intestinal microflora,and Sorenson's pairwise similarity coefficient,was established.The four major clinical recipes of traditional Chinese drugs for Pi-deficiency in rats,especially at their medium dose(equivalence to the clinical dose),produced more pronounced recovery activities in Pi-deficient rats,while higher doses of these recipes did not show a better therapeutic effect but some toxic effects such as perturbation deterioration of intestinal microflora.CONCLUSION:Both fingerprint analysis and identified marker can show Pi-deficiency in rats and its difference after treatment.The identified molecular marker may be applied in screening for the active compounds both in relative traditional Chinese drugs and in pharmacodynamic study of Pi-deficiency in rats.
基金supported by the Zhejiang Provincial Key Technology Innovation Team(No.2010R50028)the Hangzhou Science and Technology Development Program(No.20110232B11),China
文摘Chlorpyrifos is a widely used insecticide in recent years,and it will produce adverse effects on soil when applied on crops or mixed with soil.In this study,nested polymerase chain reaction（PCR） and denaturing gradient gel electrophoresis（DGGE） were combined to explore the bacterial and fungal community successions in soil treated with 5 and 20 mg/kg of chlorpyrifos.Furthermore,isolates capable of efficiently decomposing chlorpyrifos were molecular-typed using enterobacterial repetitive intergenic consensus-PCR（ERIC-PCR）.Under the experimental conditions,degradation of chlorpyrifos in soil was interpreted with the first-order kinetics,and the half-lives of chlorpyrifos at 5 and 20 mg/kg doses were calculated to be 8.25 and 8.29 d,respectively.DGGE fingerprint and principal component analysis（PCA） indicated that the composition of the fungal community was obviously changed with the chlorpyrifos treatment,and that samples of chlorpyrifos treatment were significantly separated from those of the control from the beginning to the end.While for the bacterial community,chlorpyrifos-treated soil samples were apparently different in the first 30 d and recovered to a similar level of the control up until 60 d,and the distance in the PCA between the chlorpyrifos-treated samples and the control was getting shorter through time and was finally clustered into one group.Together,our results demonstrated that the application of chlorpyrifos could affect the fungal community structure in a quick and lasting way,while only affecting the bacterial community in a temporary way.Finally,nine typical ERIC types of chlorpyrifos-degrading isolates were screened.
