Development and Characterization of Polymorphic Microsatellite Markers for Sedum sarmentosum (Crassulaceae) and Their Cross-Species Transferability.

Sedum sarmentosum is an important Chinese medicinal herb that exhibits anti-inflammatory, anti-angiogenic and anti-nociceptive properties. However, little is known about its genetic background. The first set of 14 microsatellite markers were isolated and characterized for S. sarmentosum using an SSR-enriched library. Fourteen polymorphic microsatellite markers were acquired with satisfactory amplifications and a polymorphic pattern in 48 S. sarmentosum individuals. The number of alleles ranged from 3 to 15. The observed and expected heterozygosities varied from 0.0833 to 0.8750 and 0.2168 to 0.9063, respectively. Two loci showed significant departure from the Hardy-Weinberg equilibrium. Cross-species amplification was carried out in other Sedum species. High rates of cross-species amplification were observed. The transferability value ranged from 85.7% in S. lineare to 64.3% in S. ellacombianum. These markers will be valuable for studying the genetic variation, population structure and germplasm characterization of S. sarmentosum and related Sedum species.

[Study on Identification of Three Medicinal Plant Leaves from Elaeagnus Genus by Infrared Spectroscopy].

OBJECTIVE: To study and identify the three species of dry medicinal plant leaves trom Elaeagnus genus (E. pungens, E. lanceolata and E. henryi) by Infrared Spectroscopy(IR). METHODS: Fourier transform infrared spectroscopy and second derivative infrared spectroscopy were used to study and compare the characteristics of leaves of three Elaeagnus medicinal plants. RESULTS: The IR spectra and second derivative infrared spectra of the three Elaeagnus plants leaves were similar on the whole, the intensity or ratio of intensity of some absorption peaks still had certain distinctions, and the differences of the second derivative infrared spectra were more obvious. There were only slight differences between large and small leaf type of samples of Elaeagnus lanceolata; the differences of the plant leaves of one species collected in different harvest periods were far smaller than those of others belonging to the same genus. CONCLUSION: IR can be relatively reliably used for identification of the three Elaeagnus leaves.

[HPLC characteristic fingerprints of sedi linearis herba and sedi herba].

OBJECTIVE: To study HPLC characteristic fingerprint of Sedum lineare from different harvest periods, and to compare with its related species Sedum sarmentosum. METHODS: The HPLC fingerprints of Sedum lineare from different collecting periods were established and compared with Sedum sarmentosum by the same detection method. RESULTS: Hyperin, isoquercitrin and astragaloside were identified from the HPLC fingerprint of Sedum lineare. The fingerprint of Sedum lineare growing in the same area but different environment were basically identical; while there were remarkable differences of Sedum lineare growing in the same place but from different harvest periods, with the area of most common peaks changing from little to great, and slightly different peak number. The HPLC fingerprint of the two Sedum species had four common peaks, but could be distinguished from each other. The optimal harvest period of these two species should be full-bloom stage. CONCLUSION: The established method can provide reference for identification and quality analysis of Sedum lineare.

[Comparative identification research on different extracts of Dai Medicine "Pokou" and its processed product by FTIR].

OBJECTIVE: To study identification methods of different extracts of Dai Medicine "Pokou" (the rhizome of Homalomena gigantea Engl. ) and its processed product made by immersing it in water, and provide reference for identification of the drug in further researches and applications. METHODS: FTIR technique was used for identifying the features of different extracted parts of this crude drug and its processed product. RESULTS: Compared with the crude drug, the petroleum ether-extracted parts of processed product turned out to have no obvious distinction in the FTIR. There was a large difference in the ethyl acetate-extracted parts, and the n-butanol-extracted parts also had certain discrepancy. A preliminary analysis was made on the chemical fundamentals which caused the changes in the FTIR before and after the drug's processing. CONCLUSION: The results provide an infrared spectral identification method for the drug and its applications.

[Comparative identification on Dai medicine "Pokou" and its processing product].

OBJECTIVE: To research identification methods of the Dai Medicine "Pokou" (the rhizome of Homalomena gigantea) and its processing product, and provide basis for identification of the drug in further research and application. METHODS: Macroscopic, microscopic observation and TLC and FTIR techniques were used to authenticate this raw medicine and its processing product. RESULTS: There were certain differences in the macroscopic features. The TLC result and infrared spectra of the samples had also obvious differences. The methods for identification of this raw medicine and its processing product were established, The detailed tissue and powder of this medicine were drawn. CONCLUSION: The results provided the basis for identification of the medicine and establishment of its quality standard.

[Study on microscopic observation and TLC identification of Dai medicine "Guomaguo" (Spondias pinnata)].

OBJECTIVE: To study identification methods of Dai medicine" Guomaguo", the fruit of Spondias pinnata. METHODS: Characteristic, microscopic observation and TLC idertification were used to authenticate this crude drug. RESULTS: The characters of the cross section, powder and TLC of the drug were reported, and the relevant drawings of the tissue, powder and TLC of this ethnomedicine were drawn. CONCLUSION: These results can supply evidences for the identification of the ethnomedicine in its exploitation and utilization.

[Studies on identification of Sedum emarginatum].

OBJECTIVE: To establish methods for identification of the whole plant of Sedum emarginatum Migo. METHODS: Macroscopic and microscopic observation and FTIR technique were used to authenticate this crude drug, and the identification characteristics were studied. RESULTS: The stem cross section and the whole plant powder had some notable micro-characters. The infrared spectras of the samples collected in the different habitats and seasons were very consistent with each other. CONCLUSION: The results can be used as the evidence for identification of this ethnomedicine.

[Study on pharmacognostic identifcation of Cassia siamea].

OBJECTIVE: To establish methods for identification of Dai medicine, the heartwoods and leaves of Cassia siamea. METHODS: Macroscopic, microscopic observation and TLC technique were used to anthenticate this crude drug, and the identification characteristics were studied. RESULTS: Macroscopic and microscopic identification methods and TLC characters of the ethnomedicine were re-ported, and the simple and detailed drawings of the transections of the heartwoods and leaves, as well as microscopic drawings of their powders were drawn in this paper. CONCLUSION: The results can serve as evidence for identification of the ethnomedicine in the utilization.

[RAPD analysis of 8 medicinal species of Selaginella].

OBJECTIVE: To identify 8 medicinal species of Selaginella with method of molecular marker and to explore their genetic relationship. METHODS: 8 primers were selected from 60 random primers to amplify the total DNA extracts of 17 samples with RAPD method and then the amplified were analyzed. RESULTS: A total of amplified 58 bands and RAPD map were obtained. Based on them, a Jaccard's genetic similarity matrix and a dendrogram for the 8 plants were established using SPSS 11.5 software. CONCLUSION: The method revealed distinct differences of different species and the variation of the same species of Selaginella from different habitats. It can provide with genetic proof for authentication of some species and units below species.

[Study on microscopic identification of Dai medicine "ge ga" (Zanthoylum armatum)].

The root, bark, leaf and fruit of Zanthoylum armatum are used as traditional crude drugs by Dai and some other minority nationalities in China, and the Dai drug name is "ge ga". This paper reported microscopic, physical and chemical characters of the bark and leaf The abbreviated and detailed diagrams of the transections of the bark and leaf, and the powder diagram of the bark were drawn in the study. The result showed that there were distinct microscopic characters of the transection and the powder, and these characters could be used as evidence for identification of the ethnomedicine in the exploitation and utilization.

[Study on HPLC fingerprint characteristics of Selaginella plants].

OBJECTIVE: To establish a method to analyze HPLC fingerprint characteristics of 10 plants from Selaginella. METHOD: HPLC was applied for establishment of fingerprints, which were used to evaluate and distinguish the different species of Selaginella. RESULT: The different species from Selaginella showed different HPLC fingerprint characteristic. The samples of the same species but collected in different period, different environment or different locations showed certain difference in fingerprints CONCLUSION: 2 important mutual fingerprint peaks were found in the 10 plants of Selaginella species and 5 peaks can be used as "main fingerprint peaks". The dates of these peaks can used for assessment of phylogenetic relation among species and evaluation of quality.

[Pharmacognostic identification on Dai medicine" guo sha" (Broussonetia papyrifera)].

The present paper reports descriptions and microscopical studies on Dai medicine" Guo sha" (Broussonetia papyrifera) , which is also the ethnomedicines of some minority nationalities in China. The diagram and the detailed drawing of the transection of the root, as well as microscopic drawing of its powder, were drawn in the studies. It was found that there are distinct microscopic characters of the transection and the powder which can serve as evidence for identification of the ethnomedicine.

[Investigations and studies on medicinal plants from Selaginellaceae in Hubei Province].

OBJECTIVE: To investigate the species, the distribution and the utilization of the medicinal plants from Selaginellaceae in Hubei Province. METHOD: Through field investigations and comparing the collected specimens and literatures, the classification and identification of the species in Hubei Province were studied. RESULT: The results indicated that 15 species of plants from Selaginellaceae in Hubei, including 14 medicinal and 1 newly recorded species existed. The distribution and use in folk medicine were investigated. And the morphological description of several species was appended. CONCLUSION: The results provided a basis for the exploitation and utilization of the medicinal plant resources of Selagingellaceae.

Relaxant effect of 1-butanol fraction from Elaeagnus pungens leaf through inhibiting L-type Ca2+ channel on guinea pig tracheal smooth muscle.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaf of Elaeagnus pungens thunb. (Family Elaeagnaceae) has been documented as an effective herb for the treatment of asthma and chronic bronchitis in traditional Chinese medicine. In the past years, only a few of preliminary studies reported the chemical constituents and pharmacology effects of the herb, but their action on the tracheal relaxation has not been investigated. AIM OF THE STUDY: To investigate the relaxing effect and mechanism of the extracts from Elaeagnus pungens leaves on guinea pig tracheal smooth muscle and bronchi smooth muscle cells. MATERIALS AND METHODS: Four fractions of different polarities from Elaeagnus pungens leaves were tested to the tracheal strips on the resting tension or pre-contracted by histamine (20 µM) and acetylcholine (20 µM). Inhibitory effects of the 1-butanol fraction (400mg/ml) on cumulative histamine and acetylcholine (0.2-20 µM) induced contraction were measured. In order to determine the mediators on the 1-butanol fraction effect, the relaxing effect of the 1-butanol fraction was evaluated in the absence and presence of ß-adrenoceptor antagonists (1 µM propranolol), K(+) channels-blockers (4-aminopyridine (2mM), tetraethylammonium chloride (5mM) or glibenclamide (10 µM)), the cyclooxygenase inhibitor (indomethacin, 10 µM), nitric oxide synthase inhibitor (Nω-nitro-L-arginine methyl ester, 100 µM) or L-type Ca(2+) channel inhibitor (nifedipine, 1 µM). Moreover, [Ca(2+)]i in bronchi smooth muscle cells was analyzed by measuring the fluorescence intensity with confocal system. RESULTS: 1-Butanol fraction induced the highest relaxant effect among four fractions of different polarities from Elaeagnus pungens leaves, and significantly relaxed the tracheal strip in the concentration-dependent manner on the resting tension and pre-contracted by histamine phosphate and acetylcholine. It also produced an unparallel rightward shift of the cumulative concentration-response curve of histamine or acetylcholine. Furthermore, the relaxant effect of 1-butanol fraction was not affected by propranolol, glibenclamide, tetraethylammonium chloride, 4-aminopyridine, indomethacin and Nω-nitro-L-arginine methyl ester. However, 1-butanol fraction-induced relaxation decreased after adding nifedipine. It also concentration-dependently inhibited CaCl2-induced contraction in the Ca(2+)-free, 60mM K(+)-containing solution. Additionally, [Ca(2+)]i in the BSMCs significantly reduced after administration of the 1-butanol fraction. CONCLUSIONS: The 1-butanol fraction from Elaeagnus pungens leaves resulted in a relaxation in the non-precontracted and pre-contracted tracheal strips. The relaxant effect was not related to K(+) channels, NO, cGMP or ß-adrenoceptors, but related to the inhibition of Ca(2+) influx through L-type Ca(2+) channels.