Deep chemical identification of phytoecdysteroids in Achyranthes bidentata Blume by UHPLC coupled with linear ion trap-Orbitrap mass spectrometry and targeted isolation / 中国天然药物

Achyranthes bidentata Blume is widely used as a traditional Chinese medicine with the effects of nourishing the liver and kidneys and strengthening muscles and bones. In this work, a rapid and simple strategy was developed for characterizing phytoecdysteroids by ultra-high-performance liquid chromatography coupled with liner ion trap-Orbitrap mass spectrometry using electrospray ionization in the negative mode. As a result, 47 phytoecdysteroids were unambiguously or tentatively characterized. Among them, seven known compounds were identified according to the reference standards along with molecular formula, retention time and fragmentation patterns, while others were mostly potential new compounds. Through targeted isolation, the structures of three new compounds were determined by NMR spectra, which were consistent with LC-MS characterization. The present study provides an efficient method to deeply characterize phytoecdysteroids.

Pharmacokinetics of Achyranthes bidentata on adjuvant arthritis rats by microdialysis and UHPLC-MS/MS / 中国中药杂志

To investigate the " drug-guide" effect of Achyranthes bidentata saponins( ABS) and geniposide( GE) in the treatment on adjuvant arthritis( AA) rats. A UHPLC-MS/MS method for the quantitative determination of GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa in rat blood and joint dialysate was established. After single or combined administration with ABS and GE was given to AA rat model,a microdialysis sampling method for rat joint cavity and jugular vein blood vessels was established to collect microdialysis samples. Waters Acquity HSS C_(18) column was used to separate the above four components,with mobile phase as acetonitrile-0. 1% formic acid water as mobile phase for gradient elution. ESI source was adopted for mass spectra in a negative ion scanning mode. Multiple reaction monitoring( MRM) mode was applied to detect the above four components. The methodological results showed that GE,zingibroside R1,ginsenoside Ro and chikusetsu saponin Ⅳa demonstrated a good linear relationship within the concentration ranges of 2-4 000,16-4 096,14-3 584,23-5 888 μg·L-1 respectively. The precision,accuracy,stability and matrix effect of these four ingredients reached the requirements of quantitative analysis of biological samples. The pharmacokinetic results demonstrated that the combined administration of ABS and GE( 60 mg·kg~(-1)+60 mg·kg~(-1)) can increase the degree of GE in joint cavity distribution,and the AUCjoint/AUCplasmwere twice of that of single administration of GE( 60 mg·kg~(-1)),which indicated that ABS might played a vital role in GE's distribution to joint cavity. Moreover,there was no significant difference between the distribution trend of total three ABS and GE in rats. The pharmacodynamics results showed that the combined administration of ABS and GE has stronger effects on paw swelling,arthritis index and synovial pathomorphology of AA rats than single administration of GE,which suggested that ABS might improve GE's anti-inflammatory effect in AA rats. Based on the above results,ABS has a targeting effect in increasing GE's concentration in joint cavity,with a synergy in efficacy.

Physiological responses of Catla catla larvae fed with Achyranthes aspera seed enriched diet and exposed to UV-B radiation.

The antioxidant and growth stimulating properties of seeds of Achyranthes aspera were evaluated on UV-B irradiated Catla catla (catla) larvae. Catla larvae (initial weight: 1.2 ± 0.01 mg) were fed with four different diets — D1, D2 and D3 containing 0.1, 0.25 and 0.5% seeds of A. aspera and D4, control diet for 35 days. The larvae were then exposed to UV-B radiation (80 µW/cm2) on every alternate day for 20 days. Survival, growth, tissue glutamic oxaloacetic transminase (GOT), tissue glutamate pyruvate transminase (GPT), thiobarbituric acid reactive substances (TBARS) and nitric oxide synthase (NOS) were studied in larvae on day-21 of irradiation. Significantly (P < 0.05) higher survival and average weight were found in D3 diet fed fish compared to the other groups. Survival rate was 8-16% higher in seed enriched diet fed groups, compared to the control one. Higher levels of GOT and GPT found in control diet fed larvae showed the degree of tissue damage due to UV-B exposure. Significantly (P < 0.05) lower level of GPT in D3 indicated the UV-B protective effect of the seed of A. Aspera (earlier, the presence of ecdysterone, essential fatty acids and amino acids, polyphenolic compounds, steroids, etc. has been reported from seed). TBARS which indicated the level of tissue lipid peroxidation were significantly (P < 0.05) higher in control group, compared to the other feeding schemes. NOS level was significantly (P < 0.05) higher in D2 and D3, compared to the D1 and control groups. In conclusion, supplementation of A. aspera seed (0.5%) improved the physiological condition (in terms of reduce lipid oxidation and better immune system) and gave bioprotection to catla larvae challenged with UV-B stress.

Studies on effects of Achyranthes bidentata on tongsaimai pellets main active ingredients chlorogenic acid, isoliquiritin, harpagoside and glycyrrhizin in vivo pharmacokinetics / 中国中药杂志

To study on the effects of Achyranthes bidentata on Tongsaimai pellets main active ingredients chlorogenic acid, isoliquiritin, harpagoside and glycyrrhizin in rats in vivo pharmacokinetic behaviors, a method for the simultaneous determination of chlorogenic acid, isoliquiritin, harpagoside and liquiritigenin in rat plasma was established by UPLC-MS/MS. The analysis was performed on a waters Acquity BEH C18 column (2.1 mm x 100 mm, 1.7 microm) with the mixture of acetonitrile and 0.1% formic acid/water as mobile phase, and the gradient elution at a flow rate of 0.3 mL x min(-1). The analytes were detected by tandem mass spectrometry with the electrospray ionization (ESI) source and in the multiple reaction monitoring (MRM) mode. It turned out that the analytes of Tongsaimai pellets groups C(max) and AUC(Q-infinity) values were higher than that with A. bidentata group, and the C(max) values of chlorogenic acid had significantly difference (P < 0.05), the AUC(0-infinity) values of chlorogenic acid and glycyrrhizin had significantly difference (P < 0.05); The T(max) and CL values of two groups had no significantly difference. Results showed that the established method was specific, rapid, accurate and sensitive for the studies of Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic, and A. bidentata have varying degrees of effects on Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic behaviors.

Screening of selected medicinal plants for their enzyme inhibitory potential - a validation of their ethnopharmacological uses

In present study four medicinal plants namely Valeriana wallichii, Xanthium strumarium, Achyranthes aspera and Duchesnea indica belonging to different families were collected in Khyber Pakhtunkhwa province and crude extract and subsequent fractions were analyzed for their inhibitory potential against acetylcholinesterase, butyrylcholinesterase and alpha-glucosidase enzymes. Valeriana wallichii, Xanthium strumarium and Achyranthes aspera were significantly active against cholinesterases. Chloroform and ethylacetate fractions of Valeriana wallichii exhibited significant activity against acetylcholinesterase [IC50: 61microg/ml] and butyrylcholinesterase enzymes [IC50: 58microg/ml], respectively. Similarly ethylacetate fraction of Achyranthes aspera showed significant activity against acetylcholinesterase [IC50: 61 microg/ml] and butyrylcholinesterase enzymes [IC50: 61 microg/ml], respectively. In case of alpha-glucosidase enzyme, the chloroform fraction of Xanthium strumarium exhibited significant inhibitory activity [IC50: 72 microg/ml] as compared to the standard compound acarbose [IC50: 483 microg/ml]. Duchesnea indica showed no such activities.

Effect of Achyranthes bidentata polysaccharides stimulated dendritic cells co-cultured with cytokine induced killer cells against SW480 cells / 中国中药杂志

Achyranthes bidentata polysaccharides (ABPS) was extracted from the root of A. bidentata. Dendritic cells (DC), which were stimulated with ABPS and/or tumor antigen SW480, were co-cultured with cytokine induced killer cells (CIK) to test the cytotoxic effect on colon cancer cell line SW480. Peripheral blood mononuclear cells (PBMNCs) which were separated from human peripheral blood were cultured to DC and CIK separately. (1) DC were divided into four groups: pure DC served as control group; ABPS (50 mg x L(-1)) stimulated DC served as experimental group; SW480 tumor antigen stimulated DC served as the second experimental group; ABPS (50 mg x L(-1)) and SW480 tumor antigen co-stimulated DC served as the third experimental group. Flow cytometry was used to detect the difference of the positive rate of molecules in the cell surface of DC, include CD80, CD86, CD1c, CD40, HLA-DR (6 samples for each group). (2) The four DC groups were mixed with CIK at the ratio 1:5 and acted as effect cells (DC + CIK groups), and the colon cancer cell line SW480 acted as target cells. The effect cells and the target cells were mixed together at the ratio 30: 1, 20:1 and 10:1 separately, and the CCK-8 kit was used to test the cytotoxic effect on colon cancer cell line SW480. (3) At the mixing ratio 30:1 of effect cells and target cells, ELISA was used to test the level of cytokines secretion, including IL-2, IL-12p70, IL-17 and TNF-alpha, in the liquid supernatant of every test group (3 duplication per sample). The results showed as following: (1) The positive rates of CD80, CD11c, HLA-DR, in the cell surface of DC which was co-stimulated by ABPS (50 mg x L(-1)) and SW480 tumor antigen, were obviously higher than the other DC groups (P < 0.05), and the positive rates of CD86, CD40 were obviously higher than the pure DC group (P < 0.05), and there was no remarkable difference with the other two DC groups. (2) At the mixing ratio 30:1, 20:1 and 10:1 of the effect cells and the target cells, the cytotoxic effect of ABPS stimulated DC + CIK group and SW480 tumor antigen stimulated DC + CIK group was obviously higher than DC + CIK group (P < 0.05), the cytotoxic effect of ABPS and SW480 tumor antigen co-stimulated DC + CIK group was obviously higher than all the other groups. (3) At the mixing ratio 30:1 of the effect cells and the target cells, the secretion levels of IL-12p70 and TNF-alpha in the liquid supernatant of the ABPS and SW480 tumor antigen co-stimulated DC + CIK group were obviously higher than all the other groups (P < 0.05), the secretion levels of IL-2 and IL-17 in the liquid supernatant of every test group have no remarkable difference. The cytotoxic effect of ABPS stimulated DC + CIK on SW480 was obviously increased. The cytotoxic effect of ABPS and SW480 tumor antigen co-stimulated DC + CIK group was obviously higher than all the other.

Anti-inflammatory activity of the topical preparation of Valeriana wallichii and Achyranthes aspera leaves

In vivo and in vitro screening of anti inflammatory activity of Valeriana wallichii and Achyranthes aspera leaves crude extract was performed, using standardized procedures. Methanolic crude extract topical formulation [cream] of Valeriana wallichii and Achyranthes aspera leaves [Family Valerianaceae and Amaranthaceae respectively], were screened for their anti-inflammatory activity, through "Carrageenan induced hind paw edema" test, for their effect on the acute and chronic phase inflammation models in male Wistar rats. Methanolic extract and its fractions were also evaluated for their in vitro anti-inflammatory activity using lipoxygenase inhibition assay. Leaves of Valeriana wallichii showed significant [p<0.001], dose dependant anti inflammatory activity, comparable with that of the standard, in animal model. The ethyl acetate fraction of Valeriana wallichii also showed considerable [IC 50=73 +/- 0.36] in vitro antiinflammatory activity as compared to standard [6.11 +/- 0.02]. Similarly Achyranthes aspera leaves showed relatively weak [p>0.05] in vivo anti- inflammatory activity. However, its activity was comparable with that of standard at 10% concentration after 5 hrs of carrageenan injection. This activity was present in ethyl acetate fraction during in vitro screening [IC 50=76 +/- 0.14] as compared to that of standard [IC 50=6.11 +/- 0.02]. The combined in vitro and in vivo Antiinflammatory screening shows that the ethyl acetate fraction of the crude extract of Valeriana wallichii and Achyranthes aspera can be used for the isolation of new Anti-inflammatory lead compounds

Effect of Huogu II Formula (II) with medicinal guide Radix Achyranthis Bidentatae on bone marrow stem cells directional homing to necrosis area after osteonecrosis of the femoral head in rabbit / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of Huogu II Formula (II) with medicinal guide Radix Achyranthis Bidentatae (Ach) on bone marrow stem cells (BMSCs) homing to necrosis area after osteonecrosis of the femoral head (ONFH) frozen by liquid nitrogen in rabbit as well as to explore the mechanism of prevention and treatment for ONFH.</p><p><b>METHODS</b>The animal model of ONFH was established by liquid nitrogen frozen on the rabbit left hind leg. Forty-eight Japanese White rabbits were randomly assigned to sham-operated group, model group, Huogu II group, and Huogu II plus Ach group, with 12 rabbits in each. During the course of ONFH animal model establishment, all rabbits were subcutaneously injected with recombinant human granulocyte colony-stimulating factor [rhG-CSF, 30 μg/(kg·day) for continuous 7 days]. Meanwhile, normal saline and decoction of the two formulae were administrated by gavage, respectively. White blood cells (WBC) were counted in peripheral blood before and after injection of rhG-CSF. Materials were drawn on the 2nd and 4th weeks after model built; bone glutamine protein (BGP) and bone morphogenetic protein 2 (BMP2) levels in serum were tested. Histopathologic changes were observed by hematoxylin and eosin (HE) staining. BMP2 mRNA levels were detected with in situ hybridization (ISH) staining. 5-Bromo-2'-deoxyuridine (BrdU) and stromal cell derived factor 1 (SDF-1) were measured by immunohistochemical assay in femoral head of the left hind leg.</p><p><b>RESULTS</b>Compared with the shamoperated group, the ratio of empty lacuna, serum BGP, and SDF-1 level in the model group increased significantly, and BMP2 in both serum and femoral head decreased significantly. However, in comparison with the model group, the empty lacuna ratio of Huogu II group and Huogu II plus Ach group decreased obviously in addition to the levels of serum BGP and BMP2, and the expressions of BMP2 mRNA, BrdU, and SDF-1 increased significantly. Above changes were particularly obvious in Huogu II plus Ach group. BGP and SDF-1 on the 2nd week and empty lacuna rate and serum BMP2 level on the 4th week in Huogu II group significantly exceeded their counterparts. On the 2nd week, only in Huogu II plus Ach group that the BrdU counting rose significantly. On the 4th week, empty lacuna rate and serum BMP2 level in Huogu II plus Ach group exceeded those in Huogu II group distinctively.</p><p><b>CONCLUSIONS</b>To a certain extent, the medicinal guide Ach improves the preventive and therapeutic effects of Huogu II Formula on experimental ONFH model. The possible mechanism of this is related to its promoting effect on directional homing of BMSCs to the necrosis area.</p>

Gastroprotective effect of Achyranthes aspera Linn. leaf on rats

OBJECTIVE@#The study was aimed at evaluating the antiulcer activity of ethanolic extract of Achyranthes aspera (EEAA) leaf.@*METHODS@#The anti-ulcer assays were performed on pylorus ligation and chronic ethanol induced ulcer model. The effects of the EEAA on gastric content volume, pH, free acidity, total acidity and ulcer index were evaluated.@*RESULTS@#The percentage of ulcer protection (59.55% and 35.58%) was significantly (P < 0.001) higher in the groups treated with the high dose of EEAA (600 mg/kg), it also reduced the volume of gastric juice and total acidity whereas, gastric pH was increased significantly.@*CONCLUSIONS@#The results of this study show significant gastroprotective activity of EEAA may be due to presence of phyto-constituents like flavanoids, saponins and tannins.

A new feruloyl tyramine glycoside from the roots of Achyranthes bidentata / 中国天然药物

AIM@#To study the chemical constituents of the roots of Achyranthes bidentata Bl.@*METHODS@#The chemical constituents were isolated and purified by macroporous adsorptive resin D101, silica gel, and ODS column chromatographies and preparetive HPLC. Their structures were elucidated on the basis of 1D and 2D NMR analyses.@*RESULTS@#Two feruloyl tyramine glycosides and seven triterpenoid saponins were obtained and identified as N-trans-feruloyl-3-methoxytyramine-4'-O-β-D-glucopyranoside (1), N-trans-feruloyl-3-methoxytyra mine-4-O-β-D-glucopyranoside (2), PJS-1 (3), chikusetsusaponin IVa (4), oleanolic acid 3-O-[β-D-glucuronopy ranoside-6-O-methyl ester]-28-O-β-D-glucopyranoside (5), oleanolic acid 3-O-[β-D-glucuronopyran-oside-6-O-ethylester]-28-O-β-D-glucopyranoside (6), oleanolic acid 3-O-[β-D-glucuronopyranoside-6-O-butyl ester]-28-O-β-D-glucopyranoside (7), ginsenoside R(0) (8) and hederagenin-28-O-β-D-glucopyranosyl ester (9).@*CONCLUSION@#Compound 1 is a new feruloyl tyramine glycoside, while compounds 2 and 9 are reported from A. bidentata for the first time.

In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants

<p><b>OBJECTIVE</b>To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants.</p><p><b>METHODS</b>Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with sterile distilled water for 3-4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration.</p><p><b>RESULTS</b>Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition. Sixty five percentages of plants were established in the field.</p><p><b>CONCLUSIONS</b>The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A. aspera and A. bidentata. The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can be easily adopted for commercial large scale cultivation.</p>

Study on seed quality of Achyranthes bidentata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the seed quality of Achyranthes bidentata from different sources and establish a standard of seed quality grading.</p><p><b>METHOD</b>Seed weight and moisture content determination, germination test, purity analysis were carried out by 100 grains method, high-temperature drying, double deck filter paper and GB/T354. 3-Seed Testing for Crops methods respectively. Eight collections were planted in several field plots.</p><p><b>RESULT</b>The 1 000 grains weight of 33 collections is 0.69 to 3.54 g with a variance coefficient 24.83%, the germination rate between 19.02% to 99% with a variance coefficient 38.24%. Seed moisture content and purity showed no significantly difference. K-means cluster analysis showed that 33 collections were divided into 3 grades: the first grade including 23 collections, the second and third grade including 5 collections respectively. Eight collections showed different field growth and yield characters.</p><p><b>CONCLUSION</b>There are significant differences in seed quality of A. bidentata, so the classification management is needed in medicinal materials market. Germination test is an effective method before cultivating.</p>

Therapeutic effects of saponins from Achyranthes bidentata in SHRsp / 中国中药杂志

<p><b>OBJECTIVE</b>To study the therapeutic effects of saponins from Achyranthes bidentata (SAB) in stroke-prone sponyaneously hypertension rats (SHRsp).</p><p><b>METHOD</b>Sixty SHRsp were randomly divided into five groups: SAB 0.10, 0.20, 0.40 g x kg(-1), Huatuo Zaizao pill group (positive control group) 2.5 g x kg(-1) and pathologic group (n=10). SAB and Huatuo Zaizao pill were used through filing stomach everyday for 20 days, pathologic group was given distilled water. The effects of SAB on blood pressure, changes of nerve state, death rate, brain index and the pathologic changes of hippocampal neuron of SHRsp were observed.</p><p><b>RESULT</b>SAB could markedly decrease brain index, death rate of SHRsp after stroke. SAB could improve nerve state of SHRsp after stroke. Also SAB could prolong survival time and prevent pathologic change of hippocampal neuron of SHRsp after stroke.</p><p><b>CONCLUSION</b>SAB is advantageous of therapying the stroke in SHRsp.</p>

Research on relationship between tissue quantitative distribution of 3H-Achyranthes bidentata ecdysterone and channel-tropism of herbal drugs in mice / 中国中药杂志

<p><b>OBJECTIVE</b>To study the relationship between tissue quantitative distribution and pharmacokinetics of 3H-achyranthes bidentata ecdysterone and the channel-tropism of herbal drugs in mice.</p><p><b>METHOD</b>3H-achyranthes bidentata ecdysterone was used as a tracer agent and injected into mice by the caudal vein. In 36 hours, the contents of the tracer agent of samples involving 9 different tracing phases and organ or tissue were determined in order to observe the dynamic quantitative distribution and excretion and pharmacokinetics of 3H-achyranthes bidentata ecdysterone and to understand the channel-tropism of herbal drugs achyranthes bidentata.</p><p><b>RESULT</b>3H-achyranthes bidentata ecdysterone of same organs in different tracing phases and the contents of 3H-achyranthes bidentata ecdysterone in same tracing phases of different organs were significantly different (P<0.01). 3H-achyranthes bidentata ecdysterone was mainly distributed, in the liver, kidney, adrenal gland, small intestine and lung. The concentration-time profiles of achyranthes bidentata ecdysterone in rats injected into mice by the caudal vein were shown to fit a two-compartment open model with half-lives of (778.65 +/- 12.36) min, the elimination of achyranthes bidentata ecdysterone from plasma was found to be in accord with linear kinetics.</p><p><b>CONCLUSION</b>The above mentioned selective distribution of 3H-achyranthes bidentata ecdysterone basically coincides with the meridian affinity and zang fu selection of the traditional Chinese medicine drug Achyranthes bidentata. This study will provide a scientific basis for the channel-tropism of A. bidentata.</p>

Evaluación fotohemolítica in vitro de Cissus sicyoides L y Achyranthes aspera L / In vitro photohemolytic assessment of Cissus sicyoides L y Achyranthes aspera

Los eritrocitos son células útiles para la identificación de agentes potencialmente fototóxicos administrados por vía sistémica, así como para el estudio de los mecanismos de fototoxicidad que involucran procesos de estrés oxidativo. OBJETIVO: evaluar el efecto fotohemolítico de extractos blandos de partes aéreas de Cissus sicyoides L (Vitaceae) y Achyranthes aspera L (Amaranthaceae). MÉTODOS: se utilizó un protocolo in vitro que emplea como modelo biológico eritrocitos humanos, los que se irradian con luz ultravioleta durante 90 min para evaluar el daño en las membranas eritrocitarias, por detección de hemoglobina liberada al medio. RESULTADOS: se observó un leve grado de hemólisis, el efecto fotohemolítico fue inferior a los controles positivos. CONCLUSIONES: los extractos de las plantas se clasificaron como no irritantes, lo cual sugiere que la hemólisis observada puede ser causada por la inestabilidad de la membrana del eritrocito, debido a la presencia de diferentes metabolitos en los extractos estudiados

Erythrocytes are useful cells to identify potentially phototoxic agents provided by systemic administration as well as to study the phototoxicity mechanisms involving oxidative stress processes. OBJECTIVE: to evaluate the photohemolytic effect of soft extracts from aerial parts of Cissus sicyoides L (Vitaceae) and Achyranthes aspera L (Amaranthaceae). METHODS: an in vitro protocol using human erythrocytes as biological model; they were ultraviolet light-radiated for 90 minutes to evaluate damage in erythrocyte membranes on the basis of detected hemoglobin released into the medium. RESULTS: mild hemolysis was observed, being the photohemolytic effect lower than that of positive controls. CONCLUSIONS: the extracts from these plants were rated as non-irritating, which suggests that observed hemolysis may be caused by unstable erythrocyte membranes resulting from the existence of different metabolites in the studied extracts

Authentification of Radix Cyathule and its adulterants based on SCAR markers / 中国中药杂志

Chuanniuxi (Radix Cyathule) is one of the most important geo-herb in Sichuan province, which adulterants are Hongniuxi (Cyathula capitata) and Huainiuxi (Achyranthes bidentata). In this paper Chuanniuxi and its adulterants were identified by SCAR markers. Nineteen populations from Tianquan, Baoxin, Huili and Jinkouhe were collected and their RAPD fingerprints were established. Based on the RAPD patterns, two polymorphic bands F300 and F500 were selected, recycled, cloned and sequenced. According to the sequences two pairs of sequence characterized amplified regions (SCAR) primers were designed and used to amplify all materials to prove the efficiency of identification of the different populations. Chuanniuxi and Huiniuxi could be distinguished by the primer SC-320, Chuanniuxi and Hongniuxi could be distinguished by the primer SC-495. Combining the two SCAR markers, Chuanniuxi, Hongniuxi and huainiuxi could be identified effectively and quickly.

Effect of phosphorus on copper tolerance in Achyranthes bidentata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of phosphorus on copper tolerance in Achyranthes bidentata.</p><p><b>METHOD</b>A PVC pipe experiment was conducted to study the interactive effects of phosphorus (P) and copper (Cu), on growth, elemental accumulation and chemical constituents of A. bidentata. Two levels of elemental P were applied at 0 (P0) and 100 ( P100) mg x kg(-1) soil with 5 levels of Cu at 0 (Cu0), 100 (Cu100), 200 (Cu200), 200 (Cu400), 200 (Cu600) mg x kg(-1) soil, respectively.</p><p><b>RESULT AND CONCLUSION</b>The biomass production between different Cu treatments, phosphorus treatment showed significant differences. The biomass reached the maximum value as the concentration of Cu and P was 100 mg x kg(-1). Low concentration of Cu improved the growth of A. bidentata. The growth was blocked as Cu concentration reached 200 mg x kg(-1) in soil, however the contents of oleanolic acid and ecdysterone in roots of A. bidentata had not influenced by Cu. P could improved the copper tolerance in A. bidentata and increased root yield. The Cu concentration in soil of the cultivation bases must be below 200 mg x kg(-1) in order to produce good quality of medicinal material.</p>

Determination of 5-hydroxymethyl-furfural in root of Achyranthes bidentata expressing different degree of floating sugar / 中国中药杂志

<p><b>OBJECTIVE</b>To study the relation between the content of 5-hydroxymethyl-furfural (5-HMF) and the degree of floating sugar in root of Achyranthes bidentata.</p><p><b>METHOD</b>An HPLC method was applied with a Waters Symmetry C18 3.9 mm x 150 mm (5 microm) column by using methanol-water (12:88) as the mobile phase at a flow rate of 1.0 mL x min(-1) and a UV detection of 280 nm.</p><p><b>RESULT</b>Along with the degree's deepening of floating sugar, the content of 5-HMF varied with the different shades of the sample. The content was 10 times higher in the black sample (highest degree of flowing suger) than that in the yellowish sample (normal). The concentrations of 5-HMT in five yellowish samples of roots of A. bidentata were 0.162 mg x g(-1) to 0.332 mg x g(-1).</p><p><b>CONCLUSION</b>The content increasing of 5-HMF in the root of A. bidentata was related to the degree of flowing sugar.</p>

Effect of medium components on callus induction and contents of polysaccharides in calli from Achyranthes bidentata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of medium components on the callus induction and the contents of polysaccharides in calli from Achyranthes bidentata.</p><p><b>METHOD</b>Leaves and stems were selected as explants. The effects of six kinds of factors including basal culture medium, carbon source, 2,4-D, 6-BA, TDZ, CH on the callus induction and the contents of ABPS in calli on the high growth point were studied by orthogonal design method. The data were analyzed with range analysis and variance analysis.</p><p><b>RESULT</b>To leaf, the optimal medium of callus induction was B5 with 2 mg x L(-1) 2,4-D, 0.5 mg x L(-1) 6-BA, 30 g x L(-1) glucose and 1 g x L(-1) CH; to stem, the optimal one was B5 with2 mg x L(-1) 2,4-D, 1 mg x L(-1) 6-BA, 30 g x L(-1) glucose and 0.5 g x L(-1) CH. In order to obtain higher contents ABPS, to leaf calli, the optimal medium was LS with 1 mg x L(-1) 6-BA, 1 mg x L(-1) TDZ and 30 g x L(-1) sucrose; to stem calli, the optimal one was LS with 1 mg L(-1) 2,4-D, 0.5 mg x L(-1) 6-BA, 1 mg x L(-1) TDZ and 30 g x L(-1) glucose.</p><p><b>CONCLUSION</b>The optimal media of callus induction were established with stems and leaves of A. bidentata as explants and with a view to an industrial production of polysaccharides by tissue and cell culture.</p>

Effects of chemical constituent in roots of Achyranthes Bidentata / 中国中药杂志

The difference of ingredients between the roots from the SP1 plants of Achyranthes Bidentata carried by satellite and the control's were evaluated in this study. The techniques of ultraviolet-visible spectroscopy (UVS), infrared spectroscopy (IR), Xray diffraction (XRD) and SDS-PAGE were used to analyse the chemical constituent in the root of A. bidentata. The results demonstrated that the UVS, IR, XRD and protein fingerprints of the roots from A. bidentata were distinct with special characters. The difference of the IR, XRD and protein fingerprints could be discriminated the satellite plants roots from those of the control, however, there were no difference of the UVS fingerprints between the satellite plants roots and the control. This indicated that the kinds of chemical ingredients were not different between the two groups, but the contents of some chemical ingredients deceased in SP1 plants of A. bidentata carried by satellite.