ABSTRACT
Anemone flaccida Fr. Schmidt is a perennial medicinal herb that contains pentacyclic triterpenoid saponins as the major bioactive constituents. In China, the rhizomes are used as treatments for a variety of ailments including arthritis. However, yields of the saponins are low, and little is known about the plant's genetic background or phytohormonal responsiveness. Using one-quarter of the 454 pyrosequencing information from the Roche GS FLX Titanium platform, we performed a transcriptomic analysis to identify 157 genes putatively encoding 26 enzymes involved in the synthesis of the bioactive compounds. It was revealed that there are two biosynthetic pathways of triterpene saponins in A. flaccida. One pathway depends on β-amyrin synthase and is similar to that found in other plants. The second, subsidiary ("backburner") pathway is catalyzed by camelliol C synthase and yields β-amyrin as minor byproduct. Both pathways used cytochrome P450-dependent monooxygenases (CYPs) and family 1 uridine diphosphate glycosyltransferases (UGTs) to modify the triterpenoid backbone. The expression of CYPs and UGTs were quite different in roots treated with the phytohormones methyl jasmonate, salicylic acid and indole-3-acetic acid. This study provides the first large-scale transcriptional dataset for the biosynthetic pathways of triterpene saponins and their phytohormonal responsiveness in the genus Anemone.
Subject(s)
Anemone , Genetics , Metabolism , Biosynthetic Pathways , Genetics , Cytochrome P-450 Enzyme System , Genetics , Metabolism , Gene Expression Profiling , Gene Expression Regulation, Plant , Glycosyltransferases , Genetics , Metabolism , Oleanolic Acid , Metabolism , Plant Growth Regulators , Pharmacology , Plant Proteins , Genetics , Metabolism , Plants, Medicinal , Rhizome , Genetics , Metabolism , Saponins , Metabolism , Triterpenes , MetabolismABSTRACT
This study was purposed to investigate the colony formation of high-proliferative potential colony-forming units (CFU-HPP) from bone marrow-derived hematopoietic cells of psoriatic patients and p16 gene promotor methylation in CFU-HPP cells, and to explore the relationship between the colony formation and the methylation status of p16 gene promoter. Bone marrow-derived mononuclear cells from psoriatic patients and normal controls were separated by density gradient centrifugation, and were cultured in methycellulose semi-solid culture medium with SCF, GM-CSF, IL-3 and IL-6 for 14 days to measure the colonies of CFU-HPP. The CFU-HPP colony cells were collected and methylation status of p16 gene promoter of CFU-HPP cell DNA modified with sodium bisulfite was detected by the methylation-specific polymerase chain reaction (MSP). The results showed that in methycellulose semi-solid culture system, the number and the size of CFU-HPP colonies of bone marrow of psoriatic patients were all significantly less than that of normal controls, the positive frequency of p16 gene promoter methylation in CFU-HPP cells was lower than that in CFU-HPP colony cells of normal controls. It is concluded that the colony formation capability of CFU-HPP from bone marrow hematopoietic progenitor cells in psoriatic patients is lower than that in normal controls, and the lower positive frequency of P16 gene promoter methylation in CFU-HPP cells perhaps closely correlated with lower CFU-HPP colony-forming capability.
Subject(s)
Humans , Cell Proliferation , Colony-Forming Units Assay , DNA Methylation , Genes, p16 , Hematopoietic Stem Cells , Pathology , Promoter Regions, Genetic , Genetics , Psoriasis , Genetics , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To assess the effect of stapes surgery on advanced otosclerosis (AO).</p><p><b>METHODS</b>In 300 cases randomly collected of otosclerosis in 1970 to 1999, 68 cases (77ears) were selected for retrospective analysis, which met the criteria of advanced otosclerosis (mixed deafness, with bone-conduction levels exceeding 40 dB and air-conduction levels exceeding 70 dB in 500 - 2000 Hz).</p><p><b>RESULTS</b>The air-conduction of sixty-eight cases (77 ears) were from 77.32 dB to 53.7 dB (500-2000 Hz) after operation, mean decreased 23.62 dB. Of 71 ears (92.21%) obtained air-conduction improved over 10 dB, of 46 ears (59.74%) gained A-B Gap closure in 10 dB. Air-conduction were from 79.01 dB to 58.23 dB (500 - 4000 Hz) after operation and mean decreased 20.78 dB. Of 68 ears (88.31%) obtained air-conduction improved over 10 dB, of 32 ears (41.56%) gained A-B gap closure in 10 dB. After followed up 5-25 years, of 67 ears were retained stabilization, of 28 ears obtained more improve than postoperation. But 4 ears had drop (all was stapes-elevation and re-improvement after them was reviewed).</p><p><b>CONCLUSION</b>Stapes surgery is effective operation to cured otosclerosis, advanced otosclerosis or far-advanced otosclerosis had greater help to improved hearing.</p>