Selaginellaceae: traditional use, phytochemistry and pharmacology / Selaginellaceae: uso tradicional, fitoquímica y farmacología

Selaginella is the only genus from Selaginellaceae, and it is considered a key factor in studying evolution. The family managed to survive the many biotic and abiotic pressures during the last 400 million years. The purpose of this review is to provide an up-to-date overview of Selaginella in order to recognize their potential and evaluate future research opportunities. Carbohydrates, pigments, steroids, phenolic derivatives, mainly flavonoids, and alkaloids are the main natural products in Selaginella. A wide spectrum of in vitro and in vivo pharmacological activities, some of them pointed out by folk medicine, has been reported. Future studies should afford valuable new data on better explore the biological potential of the flavonoid amentoflavone and their derivatives as chemical bioactive entities; develop studies about toxicity and, finally, concentrate efforts on elucidate mechanisms of action for biological properties already reported.

Selaginella es el único género de Selaginellaceae, y se considera un factor clave en el estudio de la evolución. La familia logró sobrevivir a las muchas presiones bióticas y abióticas durante los últimos 400 millones de años. El propósito de esta revisión es proporcionar un resumen actualizado de Selaginella para reconocer su potencial y evaluar futuras oportunidades de investigación. Los hidratos de carbono, pigmentos, esteroides, derivados fenólicos, principalmente flavonoides, y alcaloides son los principales productos naturales en Selaginella. Se ha informado un amplio espectro de actividades farmacológicas in vitro e in vivo, algunas de ellas señaladas por la medicina popular. Los estudios futuros deberían proporcionar datos nuevos y valiosos para explorar mejor el potencial biológico de la amentoflavona flavonoide y sus derivados como entidades bioactivas químicas; desarrollar estudios sobre la toxicidad y, finalmente, concentrar los esfuerzos en dilucidar los mecanismos de acción para las propiedades biológicas ya informadas.

Fractions of Selaginella convoluta (Arn) Spring (Selaginellaceae) attenuate the nociceptive behavior events in mice / Frações de Selaginella convoluta (Ar) Spring (Selaginellaceae) atenuam a resposta nociceptiva em camundongos

Abstract Selaginella convoluta (Arn.) Spring is a species popularly known as "jericó", and used in folk medicine as analgesic and anti-inflammatory. This study aimed to investigate in mice the antinociceptive and anti-inflammatory activities of the hexane (Sc-Hex) and chloroform (Sc-CHCl3) fractions (100, 200 and 400 mg/kg) obtained by partition of crude ethanol extract from S. convoluta. The preliminary phytochemical analysis of the fractions was performed. Antinociceptive activity was evaluated by writhing, formalin and hot-plate tests. Anti-inflammatory activity was evaluated using carrageenan-induced pleurisy. The rota-rod test was used to evaluate motor coordination. Preliminary phytochemical screening showed that the Sc-Hex and the Sc-CHCl3 were positive for the presence of flavonoids, anthracene derivatives, quinones, triterpenes and steroids. Inhibition of writhing was observed for fractions tested. The Sc-Hex at all doses tested was effective in reducing the nociceptive behavior produced by formalin only in the second phase. However, the Sc-CHCl3 decreased the paw licking time in the first and second phases. In the hot plate no significant effect was observed for any fraction. In the rota-rod test, treated mice did not demonstrate any significant motor performance changes. In the carrageenan-induced pleurisy, Sc-CHCl3 (200 mg/kg) reduced cell migration to the pleural cavity. These results reveal the antinociceptive properties of S. convoluta , which support, in part, its traditional use, since the fractions did not presented significant activity in the inflammatory response profile. We further verify that this antinociceptive effect could be by activation of nociceptive peripheral pathway.

Resumo Selaginella convoluta é uma espécie popularmente conhecida como "jericó", e usada na medicina popular como analgésica e anti-inflamatória. Este estudo teve como objetivo investigar em camundongos as atividades antinociceptiva e anti-inflamatória das frações hexânica (Sc-Hex) e clorofórmica (Sc-CHCl3) (100, 200 e 400 mg/kg) obtidas por partição do extrato etanólico bruto de S. convoluta . A análise fitoquímica preliminar das frações foi realizada. A atividade antinociceptiva foi avaliada pelos testes de contorções abdominais, formalina e placa quente. A atividade anti-inflamatória foi avaliada usando pleurisia induzida por carragenina. O teste do rota-rod foi utilizado para avaliar a coordenação motora. A triagem fitoquímica preliminar mostrou que Sc-Hex e Sc-CHCl3 apresentaram reação positiva para a presença de flavonoides, derivados antracênicos, quinonas, triterpenos e esteroides. A inibição das contorções foi observada para as frações testadas. Sc-Hex em todas as doses testadas foi efetivo na redução do comportamento nociceptivo produzido pela formalina somente na segunda fase. No entanto, Sc-CHCl3 diminuiu o tempo de lambida da pata na primeira e segunda fases. Na placa quente, nenhum efeito significativo foi observado para qualquer fração. No teste do rota-rod os camundongos tratados não demonstraram mudanças significativas na coordenação motora. Na pleurisia induzida por carragenina, Sc-CHCl3 (200 mg/kg) reduziu a migração celular para a cavidade pleural. Estes resultados revelam a propriedade antinociceptiva de S. convoluta, justificando, em parte, seu uso tradicional, uma vez que os extratos não apresentaram atividade significativa no perfil de resposta inflamatória. Verificamos ainda que esse efeito antinociceptivo mostrou-se ligado à ativação da via periférica nociceptiva.

Paleo-polyploidization in Lycophytes / 基因组蛋白质组与生物信息学报·英文版

Lycophytes and seed plants constitute the typical vascular plants. Lycophytes have been thought to have no paleo-polyploidization although the event is known to be critical for the fast expansion of seed plants. Here, genomic analyses including the homologous gene dot plot analysis detected multiple paleo-polyploidization events, with one occurring approximately 13-15 million years ago (MYA) and another about 125-142 MYA, during the evolution of the genome of Selaginella moellendorffii, a model lycophyte. In addition, comparative analysis of reconstructed ancestral genomes of lycophytes and angiosperms suggested that lycophytes were affected by more paleo-polyploidization events than seed plants. Results from the present genomic analyses indicate that paleo-polyploidization has contributed to the successful establishment of both lineages-lycophytes and seed plants-of vascular plants.

First Report of Agrobacterium rhizogenes-induced Hairy Root Formation in Selaginella bryopteris: a Pteridophyte Recalcitrant to Genetic Transformation

Abstract we report A. rhizogenes-induced hairy root formation in S. bryopteris, a medicinally and commercially important plant. A. rhizogenes strain LBA1334 co-cultivated with explants (root, rhizophore, stem portion near the root, and stem with intact fronds) for 24 and 48 h after transformation for induction of hairy roots. The induction of hairy root was observed after 6 days of infection in case of 48 h co-cultivation only. PCR with rolA and virC gene specific primers confirmed the induced hairy roots were due to Ri T-DNA integration and not due to contaminating A. rhizogenes. The root network as explants showed the maximum transformation efficiency. We tested different media like MS, SHFR (Stage Hog Fern Root) and KNOP's during transformation for hairy root induction. The SHFR based media showed good response in transformation as well as propagation. Further, transformation efficiency was enhanced by addition of TDZ (2 mg/L) and Bevistin (0.1%) in SHFR media. The present work would be helpful in hairy roots-based in vitro production of secondary metabolites and on aspect of functional genomics of S. bryopteris.

Effect of Pyunkang-tang on Inflammatory Aspects of Chronic Obstructive Pulmonary Disease in a Rat Model

We investigated the anti-inflammatory effect of Pyunkang-tang extract (PGT), a complex herbal extract based on traditional Chinese medicine that is used in Korea for controlling diverse pulmonary diseases, on cigarette smoke-induced pulmonary pathology in a rat model of chronic obstructive pulmonary disease (COPD). The constituents of PGT were Lonicerae japonica, Liriope platyphylla, Adenophora triphilla, Xantium strumarinum, Selaginella tamariscina and Rehmannia glutinosa. Rats were exposed by inhalation to a mixture of cigarette smoke extract (CSE) and sulfur dioxide for three weeks to induce COPD-like pulmonary inflammation. PGT was administered orally to rats and pathological changes to the pulmonary system were examined in each group of animals through measurement of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels in bronchoalveolar lavage fluid (BALF) at 21 days post-CSE treatment. The effect of PGT on the hypersecretion of pulmonary mucin in rats was assessed by quantification of the amount of mucus secreted and by examining histopathologic changes in tracheal epithelium. Confluent NCI-H292 cells were pretreated with PGT for 30 min and then stimulated with CSE plus PMA (phorbol 12-myristate 13-acetate), for 24 h. The MUC5AC mucin gene expression was measured by RT-PCR. Production of MUC5AC mucin protein was measured by ELISA. The results were as follows: (1) PGT inhibited CSE-induced pulmonary inflammation as shown by decreased TNF-α and IL-6 levels in BALF; (2) PGT inhibited the hypersecretion of pulmonary mucin and normalized the increased amount of mucosubstances in goblet cells of the CSE-induced COPD rat model; (3) PGT inhibited CSE-induced MUC5AC mucin production and gene expression in vitro in NCI-H292 cells, a human airway epithelial cell line. These results suggest that PGT might regulate the inflammatory aspects of COPD in a rat model.

Cloning and expression analysis in resurrection process of trehalose-6-phosphate synthase gene from Selaginella tamariscina / 中国中药杂志

Selaginella tamariscina is a typical resuscitation medicinal plant with extreme drought tolerance. Trehalose plays an important role in the resurrection process, and the trehalose-6-phosphate synthase(TPS) is the key enzyme to synthesize trehalose in plants. In this study, the sequence of TPS was obtained by splicing from the transcriptome data of S. tamariscina. After the synthesis of cDNA based on the template of total RNA, the sequence was cloned by RT-PCR for verification and then analyzed by bioinformatics methods. The results indicated that the full-length coding sequence of StTPS was 2 799 bp (GenBank accession no. MH155231), and the encoded protein contained 932 amino acids. StTPS could be located in the chloroplastid according to subcellular localization prediction. There were two conserved domains belonging to glycogen phosphorylase glycosyltransferase (GPGTF) family but no signal peptide or transmembrane domain in StTPS. The expression of StTPS was determined by qRT-PCR and the variation of trehalose content was measured by HPLC-ELSD during the resurrection process of S. tamariscina. Meanwhile, the correlation between them was analyzed. The results showed that both the expression level of StTPS and the trehalose content increased associated with the extension of dehydration time, and declined associated with the extension of rehydration time which proved a significant positive correlation between the StTPS expression level and the trehalose content. The results suggested that the StTPS probably plays a central role in recovery process in S. tamariscina.

In vivo antileishmanial activity and chemical profile of polar extract from Selaginella sellowii

The polar hydroethanolic extract from Selaginella sellowii(SSPHE) has been previously proven active on intracellular amastigotes (in vitro test) and now was tested on hamsters infected with Leishmania (Leishmania) amazonensis (in vivo test). SSPHE suppressed a 100% of the parasite load in the infection site and draining lymph nodes at an intralesional dose of 50 mg/kg/day × 5, which was similar to the results observed in hamsters treated with N-methylglucamine antimonate (Sb) (28 mg/Kg/day × 5). When orally administered, SSPHE (50 mg/kg/day × 20) suppressed 99.2% of the parasite load in infected footpads, while Sb suppressed 98.5%. SSPHE also enhanced the release of nitric oxide through the intralesional route in comparison to Sb. The chemical fingerprint of SSPHE by high-performance liquid chromatography with diode-array detection and tandem mass spectrometry showed the presence of biflavonoids and high molecular weight phenylpropanoid glycosides. These compounds may have a synergistic action in vivo. Histopathological study revealed that the intralesional treatment with SSPHE induced an intense inflammatory infiltrate, composed mainly of mononuclear cells. The present findings reinforce the potential of this natural product as a source of future drug candidates for American cutaneous leishmaniasis.

Molecular characteristics of two Phi glutathione S-transferases in Selaginella moellendorffii / 生物工程学报

Glutathione S-transferase (GST) is important in plants to resist various stresses. In this study, two Phi GST genes (SmGSTF1 and SmGSTF2) were cloned from Selaginella moellendorffii. SmGSTF1 and SmGSTF2 genes encode proteins of 215 amino acid residues. Gene expression analysis showed that the two genes were expressed in roots, stems and leaves. The recombinant SmGSTF1 and SmGSTF2 proteins were overexpressed in Escherichia coli, and purified by Ni-affinity chromatography. SmGSTF1 and SmGSTF2 had the catalytic activity towards 1-Chloro-2,4-Dieitrobenzene, 4-Chloro-7-nitro-1,2,3-benzoxadiazole (NBD-Cl), and 4-Nitrobenzyl chloride substrates. SmGSTF1 also had the activity towards Fluorodifen and Cumyl hydroperoxide (Cum-OOH), whereas SmGSTF2 not. The enzyme kinetics analysis showed that SmGSTF1 and SmGSTF2 had high affinity towards glutathione, and low affinity towards 1-Chloro-2, 4-Dieitrobenzene. The enzymatic activity of SmGSTF1 and SmGSTF2 had high catalytic activity between pH 7 and 8.5, and between 45 and 55 °C. SmGSTF1 and SmGSTF2 may have an important role in the resistance of Selaginella moellendorfii against stress.

A new selaginellin derivative from Selaginella pulvinata / 药学学报

Abstract: A new selaginellin derivative named as selaginellin S (1) was isolated from the whole plants of Selaginella pulvinata (Hook. et Grev.) Maxim. (Selaginellaceae), together with a known one (selaginellin M, 2). Compounds 1 and 2 were separated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were determined on the basis of extensive spectroscopic analysis including IR, MS, 1D and 2D NMR experiments, as well as ECD calculations. Compound 1 is a key intermidiant in the biosynthesis pathway of selaginellins. Compound 2 is first reported in this plant.

Flavonoids from Selaginella uncinata / 中国中药杂志

In the current study, nine flavonoids were isolated and purified from 75% ethanol extract of Selaginella uncinata (Desv.) Spring by column chromatographic techniques over macroporous resin, polyamide, silica gel, Sephadex LH-20 and pre-HPLC. On the basis of their physico-chemical properties and spectroscopic data analyses, these compounds were elucidated as cirsimarin (1), nepitrin (2), apigenin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (3), apigenin-6-C-β-D-glucopyranosyl-8-C-α-L-arabinopyranoside (4), apigenin-7-O-β-D-glucopyranoside (5), 2,3-dihydroamentoflavone (6), 4'-O-methylamentoflavone (7), 2,3-dihydro-4'-O-methyl-amentoflavone (8), and 2,3,2",3"-tetrahydron-4'-O-methyl-robustaflavone (9). Compounds 1-5 belong to flavonoid glycosides and were isolated from the genus Selaginella for the first time.

In vitro activity of the hydroethanolic extract and biflavonoids isolated from Selaginella sellowii on Leishmania (Leishmania) amazonensis

This study is the first phytochemical investigation of Selaginella sellowii and demonstrates the antileishmanial activity of the hydroethanolic extract from this plant (SSHE), as well as of the biflavonoids amentoflavone and robustaflavone, isolated from this species. The effects of these substances were evaluated on intracellular amastigotes of Leishmania (Leishmania) amazonensis, an aetiological agent of American cutaneous leishmaniasis. SSHE was highly active against intracellular amastigotes [the half maximum inhibitory concentration (IC50) = 20.2 µg/mL]. Fractionation of the extract led to the isolation of the two bioflavonoids with the highest activity: amentoflavone, which was about 200 times more active (IC50 = 0.1 μg/mL) and less cytotoxic than SSHE (IC50 = 2.2 and 3 μg/mL, respectively on NIH/3T3 and J774.A1 cells), with a high selectivity index (SI) (22 and 30), robustaflavone, which was also active against L. amazonensis (IC50 = 2.8 µg/mL), but more cytotoxic, with IC50 = 25.5 µg/mL (SI = 9.1) on NIH/3T3 cells and IC50 = 3.1 µg/mL (SI = 1.1) on J774.A1 cells. The production of nitric oxide (NO) was lower in cells treated with amentoflavone (suggesting that NO does not contribute to the leishmanicidal mechanism in this case), while NO release was higher after treatment with robustaflavone. S. sellowii may be a potential source of biflavonoids that could provide promising compounds for the treatment of cutaneous leishmaniasis.

Diuretic effect of alkaloids fraction extracted from Selaginella lepidophylla (Hook. et Grev.) Spring / Efecto diurético de la fracción con contenido de alcaloides extraída deSelaginella lepidophylla (Hook. et Grev.) Spring

The aerial parts of Selaginella lepidophylla (Hook. et Grev.) Spring, are used in Mexican folk medicine to treat renal diseases. The aim of this study was to measure the diuretic response of an aqueous extract (200 mg/kg) and alkaloids fraction at different doses (10, 40 y 100 mg/kg) of this plant and compare it with that induced by furosemide (4 mg/kg). Extract, alkaloids fraction, furosemide and vehicle were administered orally to adult rats and the effects in sodium, potassium and water balance were measured. The extract, the alkaloids fraction and the furosemide produced important and significant increments in urinary excretion of sodium, potassium and water with respect to control group. This increment was dose dependent of the alkaloids fraction, the highest dose produced a major effect. Potassium excretion increased but it was less than the one induced by furosemide. These results suggest that the aqueous extract and rich fraction in alkaloids from S. lepidophylla induce diuretic response.

Las partes aéreas de Selaginella lepidophylla (Hook. et Grev.) Spring, son usadas en la medicina tradicional mexicana para tratar enfermedades renales. El objetivo del estudio fue medir la respuesta diurética de un extracto acuoso (200 mg/kg) y de diferentes dosis de la fracción de alcaloides (10, 40 y 100 mg/kg) de esta planta y compararla con la inducida por furosemida (4 mg/kg). El extracto, la fracción de alcaloides, la furosemida y el vehículo fueron administrados por vía oral a ratas adultas y se midieron los efectos en el balance de sodio, potasio e hídrico. El extracto, la fracción de alcaloides y la furosemida produjeron importantes y significativos incrementos en la excreción urinaria de sodio, potasio y agua con respecto al grupo testigo. Este incremento fue dependiente de la dosis de la fracción de alcaloides, la dosis más alta produjo el mayor efecto. El incremento en la excreción de potasio fue menor al de furosemida. Los resultados sugieren que el extracto acuoso y la fracción rica en alcaloides de S. lepidophylla inducen una respuesta diurética.

Sinensioside A, a new sesquilignan glycoside from Selaginella sinensis / 中国天然药物

AIM@#To investigate the chemical constituents of Selaginella sinensis (Desv.) Spring.@*METHODS@#Chromatographic separations on Diaion HP-20, silica gel, and Sephadex LH-20 were used. The structures of the isolates were elucidated on the basis of spectroscopic analysis, as well as chemical methods.@*RESULTS@#Eight compounds were obtained and their structures were identified as sinensioside A (1), syringaresinol-4- O-β-D-glucopyranoside (2), (+)-medioresinol-4-O-β-D-glucopyranoside (3), pinoresinol-4, 4'-di-O-β-D-glucopyranoside (4), quercetin (5), eucomic acid (6), shikimic acid (7), and 2, 3-dihydroamentoflavone (8).@*CONCLUSION@#Compound 1 is a new dihydrobenzofuran sesquilignan glycoside from Selaginella sinensis.

Protection effect of amentoflavone in Selaginella tamariscina against TNF-alpha-induced vascular injury of endothelial cells / 药学学报

This study is to observe the protection effect of amentoflavone (AMT) in Selaginella tamariscina against TNF-alpha-induced vascular inflammation injury of endothelial cells. On the basis of TNF-alpha induced human umbilical vein endothelial cell, observe the influence of AMT on endothelial active factor, the contents of SOD and MDA, the protein expression of vascular endothelial adhesion molecules and inflammatory factor; study the effect of its common related signal pathways such as NF-kappaB; research the effect of AMT against TNF-a induced human umbilical vein endothelial cell injury by means of MTT, ELISA, Western blotting and the cell immunofluorescence. The results showed that AMT could increase the content of NO and decrease the levels of VCAM-1, E-selectin, IL-6, IL-8 and ET-1; enhance the activity of SOD, reduce the content of MDA; downregulate the protein expressions of VCAM-1, E-selectin, NF-kappaBp65 and up-regulate IkappaBalpha, attenuate the NF-kappaBp65 transfer to cell nucleus. AMT has the effect of protect vascular endothelial and maybe via the signal pathway of NF-kappaB to down-regulate the inflammation factor and oxidative damage factor of downstream.

Fenóis totais, flavonoides totais e atividade antioxidante de Selaginella convoluta (Arn. ) Spring (Selaginellaceae) / Total phenols, total flavonoids and antioxidant activity of Selaginella convoluta (Arn. ) Spring (Selaginellaceae)

Selaginella convoluta é uma espécie conhecida no Nordeste do Brasil como “jericó”, e bastante utilizada na medicina popular para tratamento de doenças. Este estudo teve como objetivo determinar o teor de compostos fenólicos e avaliar a atividade antioxidante in vitro do extrato etanólico e das frações obtidas por partição de S. convoluta. O conteúdo de fenóis totais foi determinado pelo método de Folin-Ciocalteu. O teor de flavonoides totais também foi avaliado. A atividade antioxidante dos extratos foi avaliada usando o método do sequestro do radical DPPH e inibição da auto-oxidação do sistema β-caroteno-ácido linoleico e comparada com os compostos de referência ácido ascórbico, BHA, BHT, quercetina e pirogalol. O conteúdo fenólico total foi de 209,90 ± 19,84 e 61,13 ± 2,50 mg equivalente de ácido gálico/g para os extratos AcOEt e EEB, respectivamente. O conteúdo de flavonoides totais foi de 155,70 ± 6,21 e 62,13 ± 4,10 para os dois extratos, respectivamente. Os extratos AcOEt e EEB apresentaram boas atividades antioxidantes. BHA foi o antioxidante mais efetivo, com um valor de IC50 de 1,62 ± 0,69 μg/mL. Os resultados obtidos mostram que S. convoluta pode ser uma boa fonte de compostos fenólicos antioxidantes. Estudos posteriores serão realizados para se chegar ao isolamento e identificação dos principais constituintes fenólicos dos extratos.

Selaginella convoluta is a species of "spike moss" (an order of pteridophytes) known in Northeast Brazil as "jericó" and widely used in popular medicine to treat several diseases. Phenolic compounds were determined in extracts of whole Selaginella convoluta plants. The total phenolics content was determined by the Folin-Ciocalteu method. Total flavonoid content was also measured. Antioxidant activities of the extracts were assayed by DPPH radical scavenging and inhibition of β-carotene-linoleic acid bleaching and compared with ascorbic acid, BHA, BHT, quercetin and pyrogallol, used as reference compounds. The total phenolics contents of ethyl acetate (EtOAc) and crude ethanol extract (CEE) were 209.90 ± 19.84 and 61.13 ± 2.50 mg of gallic acid equivalent/g, respectively. The total flavonoids contents were 155.70 ± 6.21 and 62.13 ± 4.10 mg of catechin equivalent/g for the two extracts, respectively. The EtOAc and CEE extracts exhibited good antioxidant activities. BHA was the most effective antioxidant, with an IC50 of 1.62 ± 0.69 μg/ml. The results show that S. convoluta could be a good source of antioxidant phenolics. Further research will be carried out to achieve the isolation and identification of the main phenolic constituents of the extracts.

Simultaneous determination of selaginellins and biflavones in Selaginella tamariscina and S. pulvinata by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a HPLC-DAD model for the simultaneous determination of two selaginellins (selaginellin and selaginellin B) and four biflavones (amentoflavone, sequoiaflavone, hinokiflavone and isocryptomerin) contained in 10 batches of Selaginella tamariscina and 12 batches of S. pulvinata produced in different areas.</p><p><b>METHOD</b>The analysis was performed on a Waters Cosmosil C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitril-0.1% formic acid as mobile phase in a linear gradient mode. The detection wavelength was set at 280, 337 nm. The flow rate was kept at 1.0 mL x min(-1), and the column temperature was 30 degrees C.</p><p><b>RESULT</b>The six active constituents showed significant different in content. Amentoflavone in S. tamariscina contains (5. 628-9. 184 mg x g(-1)) is more than that contained in S. pulvinata (0.823-7.131 mg x g(-1)), while selaginellin in S. pulvinata (0.123-0.593 mg x g(-1)) is more than that contained in S. tamariscina (0.067-0.133 mg x g(-1)). All the calibration curves showed good linear correlation coefficients (r > 0.9997) over the wide test ranges.</p><p><b>CONCLUSION</b>The developed HPLC-DAD method is simple, sensitive and accurate and has the good repeatability in separation, which is available for the quality control of S. tamariscina and S. pulvinata.</p>

Molecular characterizations of two dehydroascorbate reductases from Selaginella moellendorffii / 生物工程学报

Plant dehydroascorbate reductase (DHAR) is a physiologically important reducing enzyme in the ascorbate-glutathione recycling reaction. In this study, two DHARs genes (SmDHAR1 and SmDHAR2) were isolated from Selaginella moellendorffii. The SmDHAR1 and SmDHAR2 genes encode two proteins of 218 and 241 amino acid residues, with a calculated molecular mass of 23.97 kDa and 27.33 kDa, respectively. The genomic sequence analysis showed SmDHAR1 and SmDHAR2 contained five and six introns, respectively. Reverse transcription PCR revealed that the SmDHAR1 and SmDHAR2 were constitutive expression genes in S. moellendorffii. The recombinant SmDHAR1 and SmDHAR2 proteins were overexpressed in E. coli, and were purified by Ni-affinity chromatography. The recombinant SmDHAR1 showed 116-fold higher enzymatic activity towards the substrate dehydroascorbate than recombinant SmDHAR2. The recombinant SmDHAR1 showed higher thermal stability than recombinant SmDHAR2. These results indicated obvious functional divergence between the duplicate genes SmDHAR1 and SmDHAR2.

Study on HPLC fingerprint characteristics of Selaginella plants / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a method to analyze HPLC fingerprint characteristics of 10 plants from Selaginella.</p><p><b>METHOD</b>HPLC was applied for establishment of fingerprints, which were used to evaluate and distinguish the different species of Selaginella.</p><p><b>RESULT</b>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</p><p><b>CONCLUSION</b>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.</p>

Experimental study of estrogenic activities of five kinds of Chinese herbal medicines / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the estrogenic activity of several kinds of Chinese herbal medicines.</p><p><b>METHOD</b>Use zoopery and reporter gene technique to study the estrogenic activity of five Chinese herbal medicines. Zoopery: weanling female Kunming mice weight 9-12 g were administrated botanical extracts of Selaginella tamariscina, Pinus Massoniana, Corallodiscus flabellate, Dryopteris sublaeta and Leonurus heterophyllus, the positive control group with Nilestriol tablets and control group with water, respectiely. On the eighth day, the animals were sacrificed and the uteri were separated solely and weighed. Reporter gene technique: Induce the expression of reporter gene controlled by ERE and measure the activity of luciferase on cell's clear supernatant.</p><p><b>RESULT</b>The botanical extracts of S. tamariscina can increase weights of mice (P < 0.01); In the expression of reporter gene controlled by ERE, Either ERalpha or ERbeta's has estrogenic activity (P < 0.01). Follow in the zoopery we find the water part and the n-butanol part of S. tamariscina are the two active parts.</p><p><b>CONCLUSION</b>S. tamariscina and it's water part and n-butanol part have estrogenic activities, effect on ERbeta is greater than ERalpha.</p>

Effect of Selaginella combined with radiotherapy on nasopharyngeal carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the Chinese herbal medicine Selaginella-induced radiosensitization of terminal nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Totally 180 patients with NPC were divided equally into 3 groups with the same radiotherapeutic protocols. The patients in group A received radiotherapy alone, those in group B were given daily Selaginella (30 g) prepared into 50 ml decoction during the entire course of radiotherapy, and those in group C had Selaginella 30 g daily in the late course of radiotherapy.</p><p><b>RESULTS</b>The complete remission rate of nasopharyngeal primary lesions in groups B and C was significantly higher than that in group A, with also significantly higher complete remission rates of the cervical lymph nodes. The acute toxicity of the skin and mucous membrane was milder in the latter two groups, but the differences were not significant.</p><p><b>CONCLUSION</b>Selaginella may induce radiosensitization for terminal NPC and does not increase the acute toxicity of radiotherapy.</p>