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      Current approaches for the authentication of medicinal Dendrobium species and its products

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          Abstract

          DendrobiumSw., a member of the family Orchidaceae, includes approximately 1100 species distributed in different parts of the world. In China, the genus is represented by 76 species and two varieties, of which D. loddigesii, D. fimbriatum, D. chrysanthum, D. officinale(= D. candidum) and D. nobileare listed in the Chinese Pharmacopoeiaas the source material for Herba Dendrobii (Shihu). Because of increased demand and high price, Herba Dendrobii is often adulterated in the trade by other related species. Many Dendrobiumspecies are over-collected from the wild and are listed under endangered taxa in the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). Therefore, a reliable authentication method is needed to regulate the trade and help conserve the species from unscrupulous collections. This review describes the present status of using medicinal Dendrobiumin China, current approaches to authenticate medicinal Dendrobiumplants and Herba Dendrobii, and the efforts under way towards the development of DNA microarrays to facilitate differentiation/identification of genuine material in complex Chinese medicinal formulations.

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          Authentication of medicinal Dendrobium species by the internal transcribed spacer of ribosomal DNA.

          Herba Dendrobii (Shihu) is a commonly used Chinese medicine derived from the stem of several orchid species belonging to the genus Dendrobium. It is rather expensive and adulteration is frequent. Proper authentication of the medicinal species is necessary to protect consumers and support conservation measures. DNA sequences of the internal transcribed spacer 2 (ITS 2) of 16 Dendrobium species were shown to be significantly different from one another by an average of 12.4% and from non-orchids and Pholidota (an adulterant of Shihu) by 29.8% and 18.8%, respectively. The intra-specific variation among the Dendrobium species studied was only about 1%. Therefore, ITS 2 regions could be adopted as a molecular marker for differentiating medicinal Dendrobium species from one another and also from non-orchids and adulterants.
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            Pyranocoumarins Isolated fromPeucedanum praeruptorumas Differentiation Inducers in Human Leukemic HL-60 Cells

            Differentiation therapy for myeloid leukemia offers great potential as a supplement to the current treatment modalities. In the present report, we investigated if the pyranocoumarins, (+/-)-4'- O-acetyl-3'- O-angeloyl- cis-khellactone (or angular pyranocoumarin, APC) isolated from the medicinal plant Peucedanum praeruptorum Dunn, could induce human acute myeloid leukemic HL-60 cells to differentiate and elucidated the molecular mechanism(s) involved. The ability of HL-60 cells to reduce nitroblue tetrazolium (NBT) was significantly increased after APC treatment for 72 h. In these differentiating HL-60 cells, cell surface differentiation markers CD11b (for myeloid cells) and CD14 (for monocytic cells) were detected in 90.3 % and 70.1 % of the cells, respectively. The differentiation inducing effect of APC was time- and dose-dependent. Treatment with 20 microg/mL APC for 72 h inhibited cell growth by 90 % and cell cycle analysis revealed an increase in the proportion of G1 phase cells. In these growth-inhibited cells the expression of the cyclin-dependent kinase inhibitor p27 kip1, but not p21 WAF1, was up-regulated as shown by Western blotting. Differentiation inducing signal pathways were investigated and it was shown that phospho-MEK and phospho-ERK were elevated shortly after the addition of APC. Pre-incubation of the cells with MEK1 inhibitor PD98059 blocked this APC-induced differentiation. Our results suggest that APC are potent inducers of HL-60 cell differentiation along both the myelocytic and monocytic lineages and are potential agents for differentiation-treatment of leukemia.
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              Allele-specific primers for diagnostic PCR authentication of Dendrobium officinale.

              Based on rDNA ITS sequences of D. officinale and the other 37 species of Dendrobium, a pair of allele-specific diagnostic primers, TP-JB01S and TP-JB01X, were designed to authenticate D. officinale from the other species. Before the diagnostic PCR, the primer pair, P1 and P2, for amplifying the whole ITS region was used to validate template DNA and to obtain the appropriate template DNA for the diagnostic PCR. Diagnostic PCRs were performed using the diagnostic primers with the total DNAs of the original plants as a template. When the annealing temperature was raised to 66 degrees C, only the template DNA of D. officinale could be amplified whereas the diagnostic PCRs of the other Dendrobium species were all negative. The diagnostic PCRs have been repeated many times and have played an important role in authenticating the stems of D. officinale in China. Compared with the authentication method by sequencing DNA fragments, the allele-specific diagnostic PCR is not only simpler and time-saving but also practical and effective.
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                Author and article information

                Journal
                applab
                Plant Genetic Resources: Characterization and Utilization
                Plant Genet. Res.
                CABI Publishing
                1479-2621
                1479-263X
                August 2005
                February 12 2007
                August 2005
                : 3
                : 02
                : 144-148
                Article
                10.1079/PGR200578
                50217eb2-833d-4fcc-9169-177bacf65663
                © 2005
                History

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