AIM： To determine if the fraction of Nardostachysjata- mansi （N J） has the potential to ameliorate the severity of acute pancreatitis （AP）. METHODS： Mice were administered the biologically active fraction of N J...AIM： To determine if the fraction of Nardostachysjata- mansi （N J） has the potential to ameliorate the severity of acute pancreatitis （AP）. METHODS： Mice were administered the biologically active fraction of N J, i.e., the 4th fraction （N J4）, intra- peritoneally, and then injected with the stable chole- cystokinin analogue cerulein hourly for 6 h. Six hours after the last cerulein injection, the pancreas, lung, and blood were harvested for morphological examination,measurement of cytokine expression, and examination of neutrophil infiltration. RESULTS： N J4 administration attenuated the sever- ity of AP and lung injury associated with AP. It also reduced cytokine production and neutrophil infiltration and resulted in the in vivo up-regulation of heine oxy- genase-1 （HO-1）. Furthermore, NJ4 and its biologically active fraction, N J4-2 inhibited the cerulein-induced death of acinar cells by inducing HO-1 in isolated pan- creatic acinar cells. CONCLUSION： These results suggest that N J4 may be a candidate fraction offering protection in AP and N J4 might ameliorate the severity of pancreatitis by induc- ing HO-1 expression.展开更多
To promote spinosad biosynthesis by improving the limited oxygen supply during high-density fermentation of Saccharopolyspora spinosa, the open reading frame of the Vitreoscilla hemoglobin gene was placed under the co...To promote spinosad biosynthesis by improving the limited oxygen supply during high-density fermentation of Saccharopolyspora spinosa, the open reading frame of the Vitreoscilla hemoglobin gene was placed under the control of the promoter for the erythromycin resistance gene by splicing using overlapping extension PCR. This was cloned into the integrating vector pSET152, yielding the Vitreoscilla hemoglobin gene expression plasmid pSET152EVHB. This was then introduced into S. spinosa SP06081 by conjugal transfer, and integrated into the chromosome by site-specific recombination at the integration site ФC31 on pSET152EVHB. The resultant conjugant, S. spinosa S078-1101, was genetically stable. The integration was further confirmed by PCR and Southern blotting analysis. A carbon monoxide differential spectrum assay showed that active Vitreoscilla hemoglobin was successfully expressed in S. spinosa S078-1101. Fermentation results revealed that expression of the Vitreoscilla hemoglobin gene significantly promoted spinosad biosynthesis under normal oxygen and moderately oxygen-limiting conditions （P〈0.01）. These findings demonstrate that integrating expression of the Vitreoscilla hemoglobin gene improves oxygen uptake and is an effective means for the genetic improvement of S. spinosa fermentation. Saccharopolyspora spinosa, spinosad, Vitreoscilla hemoglobin, integrating vector, homologous recombination展开更多
基金Supported by The Ministry of Education,Science and Technology at Wonkwang University,No. MEST 2010-0017094
文摘AIM： To determine if the fraction of Nardostachysjata- mansi （N J） has the potential to ameliorate the severity of acute pancreatitis （AP）. METHODS： Mice were administered the biologically active fraction of N J, i.e., the 4th fraction （N J4）, intra- peritoneally, and then injected with the stable chole- cystokinin analogue cerulein hourly for 6 h. Six hours after the last cerulein injection, the pancreas, lung, and blood were harvested for morphological examination,measurement of cytokine expression, and examination of neutrophil infiltration. RESULTS： N J4 administration attenuated the sever- ity of AP and lung injury associated with AP. It also reduced cytokine production and neutrophil infiltration and resulted in the in vivo up-regulation of heine oxy- genase-1 （HO-1）. Furthermore, NJ4 and its biologically active fraction, N J4-2 inhibited the cerulein-induced death of acinar cells by inducing HO-1 in isolated pan- creatic acinar cells. CONCLUSION： These results suggest that N J4 may be a candidate fraction offering protection in AP and N J4 might ameliorate the severity of pancreatitis by induc- ing HO-1 expression.
基金supported by the National Basic Research Program of China (Grant Nos. 2012CB722301 and 2011CB111605)the National High Technology Research and Development Project of China (Grant No. 2011AA10A203)the National Natural Science Foundation of China (Grant No. 31070006)
文摘To promote spinosad biosynthesis by improving the limited oxygen supply during high-density fermentation of Saccharopolyspora spinosa, the open reading frame of the Vitreoscilla hemoglobin gene was placed under the control of the promoter for the erythromycin resistance gene by splicing using overlapping extension PCR. This was cloned into the integrating vector pSET152, yielding the Vitreoscilla hemoglobin gene expression plasmid pSET152EVHB. This was then introduced into S. spinosa SP06081 by conjugal transfer, and integrated into the chromosome by site-specific recombination at the integration site ФC31 on pSET152EVHB. The resultant conjugant, S. spinosa S078-1101, was genetically stable. The integration was further confirmed by PCR and Southern blotting analysis. A carbon monoxide differential spectrum assay showed that active Vitreoscilla hemoglobin was successfully expressed in S. spinosa S078-1101. Fermentation results revealed that expression of the Vitreoscilla hemoglobin gene significantly promoted spinosad biosynthesis under normal oxygen and moderately oxygen-limiting conditions （P〈0.01）. These findings demonstrate that integrating expression of the Vitreoscilla hemoglobin gene improves oxygen uptake and is an effective means for the genetic improvement of S. spinosa fermentation. Saccharopolyspora spinosa, spinosad, Vitreoscilla hemoglobin, integrating vector, homologous recombination
