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Objective:According to the GB/T 15000.3-2008， to develop a fucosterol certified reference material based on the project approved by Standardization Administration. Method:Fucosterol was isolated from <italic>Laminaria japonica</italic> dried thallus via 95% ethanol extraction， vacuum concentration， repeated column chromatography separation， recrystallization in petroleum ether-ethyl acetate， and residual solvent removal. Its chemical structure was identified by elemental analysis （EA）， infrared spectrum （IR）， mass spectrometry （MS）， nuclear magnetic resonance （NMR） and X-ray diffraction （XRD）. Its homogeneity， stability， and cooperative certification conducted by 8 laboratories were carried out by high performance liquid chromatography with evaporative light scattering detector. Result:For the fucosterol reference material， the certified value of purity was 99.54% with expanded uncertainty of 0.16% in confidence interval of 95%， the stability was good within 24 months storage period at 2-4 ℃， which met the technical requirements of reference material and passed the acceptance organized by Standardization Administration. Conclusion:The national standard materials of fucosterol has been successfully developed， which can be used for the determination of this component， the evaluation of detection methods， and the detection and quality control of related products.
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Abstract Lichens are traditionally divided into types such as "crustose", "foliose" and "fruticose", with different shapes and heights. Substrate, temperature and water are thought to affect lichen height, but there are few studies regarding tropical paramo lichens. Along 2015 I measured those variables in the terricolous lichens of the Cerro Buena Vista, paramo (Costa Rica, 9°33' N & 83°45' W). The measurements were taken inside the lichens and in the substrate under them, in 61 randomly located quadrats (50 x 50 cm). Lichens grew taller on (1) warmer ground, (2) wetter ground, (3) the moister Caribbean slope, and (4) the season with heavier rainfall, as expected. Apparently, atmospheric factors are more important than substrate in the determination of temperature, relative humidity and growth of lichens. Physiologically available water seems to be the main determinant of lichen vertical growth in the Buena Vista paramo.(AU)
Resumen Los líquenes se dividen tradicionalmente en morfotipos como "crustoso", "folioso" y "fruticoso", con diferentes formas y alturas. Se cree que el sustrato, la temperatura y el agua afectan la altura de los líquenes, pero existen pocos estudios con respecto a los líquenes tropicales. A lo largo del 2015, medí esas variables en los líquenes del suelo en el páramo de Cerro Buena Vista (Costa Rica, 9 ° 33'N; 83 ° 45'W). Las medidas se tomaron dentro de los líquenes y en el sustrato debajo de ellos, en 61 cuadrantes ubicados al azar (50 x 50 cm). Los líquenes fueron más altos en (1) suelo más cálido, (2) suelo más húmedo, (3) la pendiente del Caribe, que es más húmeda y (4) la temporada con lluvias más intensas, como se esperaba. Aparentemente, los factores atmosféricos son más importantes que el sustrato en la determinación de la temperatura, la humedad relativa y el crecimiento de los líquenes. El agua fisiológicamente disponible parece ser el principal determinante del crecimiento vertical de los líquenes en el páramo del Cerro Buena Vista.(AU)
Subject(s)
Tropical Climate , Tropical Ecosystem , Physical Phenomena , Lichens/growth & development , Costa RicaABSTRACT
Objective: To analyze the differences in protein of thallus of Huperzia serrata (in vitro cultures, strains A, B, and C, respectively) with different Hup A content, and to explore related enzymes that may be synthesized with Hup A accumulation. Methods: Quantitative proteomics was performed in vitro cultures of Huperzia serrata with different Hup A content using quantitative proteomics tandem mass tag (TMT) techniques, followed by differential protein analysis: GO, KEGG and other biological information analysis. Results: Strain B hadthe lowest Hup A content and strain C had the highest Hup A content, which was twice than that of B. There were 78 differential proteins between the strain B and C. Analysis of differential protein GO enrichment showed that MF accounted for 28.75%; Analysis of differential protein expression showed that three strains shared two differential proteins (P93541, Q8RXU4) in the alkaloid metabolic pathway starting from amino acids. P93541 protein was down-regulated in the low-yield strain B and up-regulated in the relatively high-yield strains A and C. The Q8RXU4 protein was up-regulated in the low-yield strain B and down-regulated in the relatively high-yield strains A and C. Conclusion: This study found that the difference in Hup A content was positively correlated with the protein expression. Two enzymes P93541 and Q8RXU4 that may be related to Hup A accumulation were analyzed, providing a basis for bioinformatics analysis of Hup A biosynthesis.
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Objective: To investigate the influence factors and dynamic varitation rule for the accumulation of huperzine A (Hup A) from thallus of Huperzia serrata, which could produce Hup A. Methods: The optimal concentration of plant growth substances for the growth of thallus of H. serrata were screened using orthogonal design method. The content of target extract from thallus of H. serrata were tested using HPLC. The kinetic curve of thallus growth was described using Logistic growth model. Results: The thallus of H. serrata could produce the most Hup A (72.45 μg/L) when cultured in 1/4 MS solid medium complemented with 12 h/d white light and the relative growth rate could reach 3 174.5%. According to the growth curve of thallus of H. serrata like "S" shape, the best successive generation date of callus culture of H. serrata was 50 d. The concentration of Hup A increased obviously after 60 d when the thallus reached the stable period and the maximal value of Hup A was 7.453 μg/g at 85 d, which meant that the rapid growth of thallus was priority to the accumulation of Hup A. Conclusion: The relational model between the accumulation of Hup A from thallus and its growth is non-growth conjugation. In addition, the growth law of thallus is consistent with Logistic growth model and the maximum specific growth rate (μmax) was 0.071 d-1.
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Objective To study the best preparation technique of compound Thallus laminaria granules volatile oil-?-cyclodextrin clathrate and examine the stability of the elathrate.Methods The orthogonal design method was used to optimize the formulation.The yeild of the volatile oil and the content of the clathrate were used as the test target.Results The best preparation process was as follows:?-cyclodextrin:compound Thallus laminaria=12:1(g?mL~(-1)),at 50～60℃for 2 hours.Making distillation twice while adding water to the first decoction, steaming 1h after boiling;A_2 B_1 was the best extraction scheme.Conclusion The preparation process can be a scheme to obtain the clathrate with high content of effective component,which can be used in the preparation of the clathrate.
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Objective To study the antithrombotic effect of polysaccharide extracted from Thallus Laminariae(PTP) on rats with endothelial injury.Methods Two kinds of endothelial injured rat models were established by injecting adrenaline and endotoxin.The wet weight of thrombus was evaluated by artery-vein loop method,the starting time of thrombus formation and the speed of blood flow were observed by mesentery microcirculation.Results In adrenaline-induced injury rats,the wet weight of thrombus was obviously decreased in high-dose PTP group and aspirin group compared with the model group.In endotoxin-induced injury rats,the starting time of thrombus formation was longer and the one-hour blood flow speed was faster in all TP groups and aspirin group than those in the model group.Conclusion PTP exerts the antithrombotic effect on rats with endothelial injury.