[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR produ...[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR products of NfEBP1 with particular restriction sites and expression vectors, pET-28 and pET-16b were digested. Ligation, transformation and selection were performed to construct the recombinant plasmids pET-28-NfEBP1 and pET-16-NfEBP1. The recombinant plasmids were transformed into E. coli BL21 using heat -shock transformation. [Results] Recombinant plasmids pET-28-NfEBP1-1188 and pET-16-NfEBP1-1188 were constructed and transformed into expressional host cells, E. coli BL21, and validated by colony PCR, sequencing and double digestion. [Conclusion] Prokaryotic expression vectors of EBP1 gene from N. fordii were successfully constructed, which laid the foundation for characterization of the gene function.展开更多
To study the chemical constituents in Bletilla ochracea Schltr. , repeated column chromatographies and preparative TLC were used for the isolation of compounds, and spectroscopic techniques （NMR, IR, UV and MS） were...To study the chemical constituents in Bletilla ochracea Schltr. , repeated column chromatographies and preparative TLC were used for the isolation of compounds, and spectroscopic techniques （NMR, IR, UV and MS） were used for their structural identification. Seven compounds, 2,7-bis （ allyloxy ） -5-methoxy-3-methyl-9, 10-dihydrophenanthrene（1）, gastrodin（2）, gastrodigenin（3）, β-sitosterol（4）, stigmasterol（5）, 4-hydroxybernzaldehyde（6）, and daucosterol（7） were obtained from the roots of B. ochracea Schltr. Compound 1 is structurally illustrated as a novel compound, and the others are isolated from the title plant for the first time.展开更多
Hepatocellular carcinoma(HCC)remains the sixth most common malignancy and the fourth leading cause of cancer death worldwide.Numerous Chinese herbs have been used to prevent and treat HCC.Periploca forrestii Schltr.,b...Hepatocellular carcinoma(HCC)remains the sixth most common malignancy and the fourth leading cause of cancer death worldwide.Numerous Chinese herbs have been used to prevent and treat HCC.Periploca forrestii Schltr.,belonging to the family of Asclepiadaceae,has been reported to have strong inhibitory efficacy against various types of cancer cells,but the mechanism is unclear.Hence,we utilized the compound-disease target network based on the network pharmacology analysis to find out the active ingredients from P.forrestii and the potential target proteins involved in hepatocellular carcinoma.The results showed that the most effective compounds in this plant might be caffeoylquinic acids and the most potential target might be GMP synthase[glutamine-hydrolyzing](PDB:2vxo).This study is of great significance to the further research of P.forrestii and provides a novel treatment strategy for HCC.展开更多
Objective：To determine the anti-inflammatory effects of an ethanol fraction of Periploca forrestii Schltr.（EFPF） and to investigate the potential mechanisms underlying in vivo and in vitro models. Methods：The anti...Objective：To determine the anti-inflammatory effects of an ethanol fraction of Periploca forrestii Schltr.（EFPF） and to investigate the potential mechanisms underlying in vivo and in vitro models. Methods：The antiinflammatory effects of EFPF were evaluated using the xylene-induced mouse ear edema and carrageenan-induced rat paw edema models in vivo. In vitro, RAW264.7 cells were exposed to 0–800 μg/m L EFPF and the cell viability was determined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay. Then cells were treated with different concentrations of EFPF（100–400 μg/m L） and stimulated with lipopolysaccharide（LPS, 1 μg/m L） for 24 h. The supernatant was analyzed for nitric oxide（NO） using the Griess reagent, and the levels of inflammatory mediators and cytokines were determined using enzyme-linked immunosorbent assays for prostaglandin E2（PGE2）, tumor necrosis factor α（TNF-α）, interleukin（IL） 6, and IL-10. The protein expressions of inducible NO synthase（i NOS）, cyclooxygenase-2（COX-2）, nuclear factor κB（NF-κB）, and mitogen-activated protein kinases（MAPKs） including extracellular signal-regulated kinase（ERK）, c-Jun N-terminal kinase（JNK）, and p38 MAPK were examined by Western blot. Results：Compared with the control group, EFPF significantly reduced mouse ear edema and rat paw edema rate（P〈0.05 or P〈0.01）. Compared with the LPS group, EFPF significantly inhibited the LPS-stimulated production of NO, PGE2, TNF-α and IL-6（P〈0.05 or P〈0.01）, and increased the IL-10 production（P〈0.05）. EFPF also significantly inhibited LPS-induced protein expressions of i NOS and COX-2, suppressed the phosphorylation and degradation of inhibitor of NF-κB-α, decreased p65 level, and inhibited the phosphorylation of p38, ERK1/2 and JNK（P〈0.05 or P〈0.01）. Conclusion：EFPF exerted anti-inflammatory effect by reducing protein expressions of i NOS and COX-2 and the production of the inflammation factors, including TNF-α, IL-6, NO and PGE2, mainly through inhibition of LPS-mediated stimulation of NF-κB and MAPK signaling pathways.展开更多
To analyze the chemical constituents of volatile oils from the fruit of Periploca forrestii Schltr.which were collected in Guizhou province.This volatile oils were extracted by simultaneous distillation extraction (SD...To analyze the chemical constituents of volatile oils from the fruit of Periploca forrestii Schltr.which were collected in Guizhou province.This volatile oils were extracted by simultaneous distillation extraction (SDE)and microwave assistant extraction(MAE) method,and ananlyzed by gas chromatography-mass spectrometry(GC-MS)method.The main chemical compounds were methyl salicylate,β-amyrin and β-amyrin acetate etc.The compounds from the volatile oils extracted with SDE and MAE are quite different.The results provide scientific proof for the further development and utilization of Periploca forrestii Schltr.展开更多
基金Supported by Research Fund of the Doctoral Program of Higher Education (200805720004)Scientific Research Foundation for Returned Scholars, Ministry of Education of China ([2009]1001)~~
文摘[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR products of NfEBP1 with particular restriction sites and expression vectors, pET-28 and pET-16b were digested. Ligation, transformation and selection were performed to construct the recombinant plasmids pET-28-NfEBP1 and pET-16-NfEBP1. The recombinant plasmids were transformed into E. coli BL21 using heat -shock transformation. [Results] Recombinant plasmids pET-28-NfEBP1-1188 and pET-16-NfEBP1-1188 were constructed and transformed into expressional host cells, E. coli BL21, and validated by colony PCR, sequencing and double digestion. [Conclusion] Prokaryotic expression vectors of EBP1 gene from N. fordii were successfully constructed, which laid the foundation for characterization of the gene function.
文摘To study the chemical constituents in Bletilla ochracea Schltr. , repeated column chromatographies and preparative TLC were used for the isolation of compounds, and spectroscopic techniques （NMR, IR, UV and MS） were used for their structural identification. Seven compounds, 2,7-bis （ allyloxy ） -5-methoxy-3-methyl-9, 10-dihydrophenanthrene（1）, gastrodin（2）, gastrodigenin（3）, β-sitosterol（4）, stigmasterol（5）, 4-hydroxybernzaldehyde（6）, and daucosterol（7） were obtained from the roots of B. ochracea Schltr. Compound 1 is structurally illustrated as a novel compound, and the others are isolated from the title plant for the first time.
文摘Hepatocellular carcinoma(HCC)remains the sixth most common malignancy and the fourth leading cause of cancer death worldwide.Numerous Chinese herbs have been used to prevent and treat HCC.Periploca forrestii Schltr.,belonging to the family of Asclepiadaceae,has been reported to have strong inhibitory efficacy against various types of cancer cells,but the mechanism is unclear.Hence,we utilized the compound-disease target network based on the network pharmacology analysis to find out the active ingredients from P.forrestii and the potential target proteins involved in hepatocellular carcinoma.The results showed that the most effective compounds in this plant might be caffeoylquinic acids and the most potential target might be GMP synthase[glutamine-hydrolyzing](PDB:2vxo).This study is of great significance to the further research of P.forrestii and provides a novel treatment strategy for HCC.
基金Supported by the National Natural Science Foundation of China(No.81460641,No.81660691)
文摘Objective：To determine the anti-inflammatory effects of an ethanol fraction of Periploca forrestii Schltr.（EFPF） and to investigate the potential mechanisms underlying in vivo and in vitro models. Methods：The antiinflammatory effects of EFPF were evaluated using the xylene-induced mouse ear edema and carrageenan-induced rat paw edema models in vivo. In vitro, RAW264.7 cells were exposed to 0–800 μg/m L EFPF and the cell viability was determined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay. Then cells were treated with different concentrations of EFPF（100–400 μg/m L） and stimulated with lipopolysaccharide（LPS, 1 μg/m L） for 24 h. The supernatant was analyzed for nitric oxide（NO） using the Griess reagent, and the levels of inflammatory mediators and cytokines were determined using enzyme-linked immunosorbent assays for prostaglandin E2（PGE2）, tumor necrosis factor α（TNF-α）, interleukin（IL） 6, and IL-10. The protein expressions of inducible NO synthase（i NOS）, cyclooxygenase-2（COX-2）, nuclear factor κB（NF-κB）, and mitogen-activated protein kinases（MAPKs） including extracellular signal-regulated kinase（ERK）, c-Jun N-terminal kinase（JNK）, and p38 MAPK were examined by Western blot. Results：Compared with the control group, EFPF significantly reduced mouse ear edema and rat paw edema rate（P〈0.05 or P〈0.01）. Compared with the LPS group, EFPF significantly inhibited the LPS-stimulated production of NO, PGE2, TNF-α and IL-6（P〈0.05 or P〈0.01）, and increased the IL-10 production（P〈0.05）. EFPF also significantly inhibited LPS-induced protein expressions of i NOS and COX-2, suppressed the phosphorylation and degradation of inhibitor of NF-κB-α, decreased p65 level, and inhibited the phosphorylation of p38, ERK1/2 and JNK（P〈0.05 or P〈0.01）. Conclusion：EFPF exerted anti-inflammatory effect by reducing protein expressions of i NOS and COX-2 and the production of the inflammation factors, including TNF-α, IL-6, NO and PGE2, mainly through inhibition of LPS-mediated stimulation of NF-κB and MAPK signaling pathways.
文摘To analyze the chemical constituents of volatile oils from the fruit of Periploca forrestii Schltr.which were collected in Guizhou province.This volatile oils were extracted by simultaneous distillation extraction (SDE)and microwave assistant extraction(MAE) method,and ananlyzed by gas chromatography-mass spectrometry(GC-MS)method.The main chemical compounds were methyl salicylate,β-amyrin and β-amyrin acetate etc.The compounds from the volatile oils extracted with SDE and MAE are quite different.The results provide scientific proof for the further development and utilization of Periploca forrestii Schltr.
