[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
Objective To investigate the effects of total flavonoid fraction(TFF) from Nervilia fordii on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in rats,and to explore their protective mechanism.Methods LPS-induce...Objective To investigate the effects of total flavonoid fraction(TFF) from Nervilia fordii on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in rats,and to explore their protective mechanism.Methods LPS-induced ALI model was established by LPS(5 mg/kg) injection via left cervical vein.Blood samples were collected from the cervical artery of all rats at 5 and 6 h after LPS challenge for arterial blood gas test and cytokines measurements,and pulmonary microvascular permeability(PMP),lung wet/dry weight ratio(W/D),and pathological features were observed.Results Phytochemical study showed that the TFF contained 67.3% of flavonoids expressed in rutin and three flavone glycosides.The TFF pretreatment(6.24 and 12.48 mg/kg) attenuated the partial arterial pressure of oxygen decline in blood significantly,and decreased the PMP and lung W/D in ALI rats.In addition,the TFF(6.24 and 12.48 mg/kg) also ameliorated the LPS-induced lung damages including alveolar edema,neutrophils infiltration,alveolar hemorrhage,and thickening of the alveolar wall.Furthermore,the treatment with the TFF(6.24 and 12.48 mg/kg) also down-regulated the levels of pro-inflammatory cytokines,such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and intercellular adhesion molecule-1(ICAM-1),and up-regulated the level of anti-inflammatory cytokine IL-10 in serum of ALI rats simultaneously.Conclusion These results suggest that the TFF could protect LPS-induced ALI in rats,which may be mediated,at least in part,by adjusting the production of inflammatory cytokines including TNF-α,IL-6,ICAM-1,and IL-10.展开更多
目的考察青天葵Nervilia fordii(Hance)Schltr.氯仿、乙酸乙酯、正丁醇、乙醇提取物抗氧化活性与其总黄酮和总酚含有量的相关性。方法 Al Cl3-HAc-Na Ac显色法和福林-酚比色法分别测定青天葵4种提取物中总黄酮和总酚含有量,通过测定DPP...目的考察青天葵Nervilia fordii(Hance)Schltr.氯仿、乙酸乙酯、正丁醇、乙醇提取物抗氧化活性与其总黄酮和总酚含有量的相关性。方法 Al Cl3-HAc-Na Ac显色法和福林-酚比色法分别测定青天葵4种提取物中总黄酮和总酚含有量,通过测定DPPH自由基清除能力、羟自由基清除能力、ABTS自由基清除能力、还原能力评价提取物抗氧化活性,SPSS17.0软件分析抗氧化活性与2种成分含有量的相关性。结果青天葵4种提取物的抗氧化活性均呈量效关系,乙酸乙酯提取物最强,而乙醇提取物最弱。其与总黄酮含有量有一定相关性(0.604<r<0.638),与总酚含有量相关性显著(r>0.977)。结论青天葵乙酸乙酯提取物可用于天然抗氧化剂开发,总酚含有量是影响其抗氧化活性的主要因素。展开更多
目的优选青天葵中两种黄酮类成分—鼠李柠檬素和鼠李秦素的最佳提取工艺条件。方法采用正交实验进行提取方法的研究,用HPLC法测定两种黄酮的含量。结果在选定的色谱条件下,RP-HPLC方法较好,结果理想。青天葵中两种黄酮的最佳提取工艺为...目的优选青天葵中两种黄酮类成分—鼠李柠檬素和鼠李秦素的最佳提取工艺条件。方法采用正交实验进行提取方法的研究,用HPLC法测定两种黄酮的含量。结果在选定的色谱条件下,RP-HPLC方法较好,结果理想。青天葵中两种黄酮的最佳提取工艺为加75%甲醇20 m l,超声提取60 m in。结论建立了准确可靠、分析速度快可同时检测青天葵中两种黄酮的分析方法,优化了质量可控的提取工艺。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.
基金Natural Science Foundation of China(30472209)Research Fund of the University of Macao(SRG013-ICMS11-CXP)
文摘Objective To investigate the effects of total flavonoid fraction(TFF) from Nervilia fordii on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in rats,and to explore their protective mechanism.Methods LPS-induced ALI model was established by LPS(5 mg/kg) injection via left cervical vein.Blood samples were collected from the cervical artery of all rats at 5 and 6 h after LPS challenge for arterial blood gas test and cytokines measurements,and pulmonary microvascular permeability(PMP),lung wet/dry weight ratio(W/D),and pathological features were observed.Results Phytochemical study showed that the TFF contained 67.3% of flavonoids expressed in rutin and three flavone glycosides.The TFF pretreatment(6.24 and 12.48 mg/kg) attenuated the partial arterial pressure of oxygen decline in blood significantly,and decreased the PMP and lung W/D in ALI rats.In addition,the TFF(6.24 and 12.48 mg/kg) also ameliorated the LPS-induced lung damages including alveolar edema,neutrophils infiltration,alveolar hemorrhage,and thickening of the alveolar wall.Furthermore,the treatment with the TFF(6.24 and 12.48 mg/kg) also down-regulated the levels of pro-inflammatory cytokines,such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and intercellular adhesion molecule-1(ICAM-1),and up-regulated the level of anti-inflammatory cytokine IL-10 in serum of ALI rats simultaneously.Conclusion These results suggest that the TFF could protect LPS-induced ALI in rats,which may be mediated,at least in part,by adjusting the production of inflammatory cytokines including TNF-α,IL-6,ICAM-1,and IL-10.
文摘目的考察青天葵Nervilia fordii(Hance)Schltr.氯仿、乙酸乙酯、正丁醇、乙醇提取物抗氧化活性与其总黄酮和总酚含有量的相关性。方法 Al Cl3-HAc-Na Ac显色法和福林-酚比色法分别测定青天葵4种提取物中总黄酮和总酚含有量,通过测定DPPH自由基清除能力、羟自由基清除能力、ABTS自由基清除能力、还原能力评价提取物抗氧化活性,SPSS17.0软件分析抗氧化活性与2种成分含有量的相关性。结果青天葵4种提取物的抗氧化活性均呈量效关系,乙酸乙酯提取物最强,而乙醇提取物最弱。其与总黄酮含有量有一定相关性(0.604<r<0.638),与总酚含有量相关性显著(r>0.977)。结论青天葵乙酸乙酯提取物可用于天然抗氧化剂开发,总酚含有量是影响其抗氧化活性的主要因素。
文摘目的优选青天葵中两种黄酮类成分—鼠李柠檬素和鼠李秦素的最佳提取工艺条件。方法采用正交实验进行提取方法的研究,用HPLC法测定两种黄酮的含量。结果在选定的色谱条件下,RP-HPLC方法较好,结果理想。青天葵中两种黄酮的最佳提取工艺为加75%甲醇20 m l,超声提取60 m in。结论建立了准确可靠、分析速度快可同时检测青天葵中两种黄酮的分析方法,优化了质量可控的提取工艺。
