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Determination of sugars in Chinese traditional drugs by CE with amperometric detection.

A simple, fast and reliable method, based on capillary zone electrophoresis with amperometric detection, for the separation and determination of sucrose, glucose, and fructose in Chinese traditional drugs, namely Astragalus Membranceus (Fish.) Bge, Angelica and Codonopsis pilosula (Franch.) Nannf. was described in this paper. A copper disk electrode was used as working electrode. The optimal conditions of separation and detection were 0.05 mol/l sodium hydroxide buffer (pH 12.7), 5 kV for the separation voltage and +0.65 V (vs. Ag/AgCl) for the detection potential. The linear ranges were from 5.0x10(-6) to 5.0x10(-4) mol/l for all three sugars. The all regression coefficients were more than 0.999. The detection limits were 1.0x10(-6) mol/l for glucose and fructose, and 4.0x10(-6) mol/l for sucrose. The method built in this paper was directly applied to the separation and determination of the three sugars in three Chinese traditional drugs without prior derivatization, and the content for every sugar in the drugs was first assayed. The assay results were satisfactory.

Angelica↗

Influences of traditional Chinese medicine on non-specific immunity of Jian Carp (Cyprinus carpio var. Jian).

The influence of traditional Chinese medicine (TCM) formulation from Astragalus Root (Radix astragalin seu Hedysari) and Chinese Angelica Root (R. Angelicae Sinensis) at a ratio of 5:1 (w/w) on non-specific immunity of Jian carp, Cyprinus carpio var. Jian was investigated. The number of NBT-positive cells in the blood and lysozyme and complement activities in the serum of Carp fed with commercial feed supplemented with 1.0% (diet 1) and 1.5% (diet 2) TCM at 10 day of post-feeding were not different from those of the control group fed with feed unsupplemented TCM 10 days post-feeding (P>0.05), but at 20 and 30 days they increased significantly (P<0.05). The values of diet 1 group and diet 2 group at 20 day and at 30 day were not significantly different (P>0.05) from each other. In addition, the TCM formula increased body weight of experimental fish by about 16.84% (diet 1) and 19% (diet 2) above that of the control group. Therefore, these data suggest that the TCM formula could elevate the function of non-specific immunity of Jian carp. The optimal dosage added to commercial carp feed was 1.0% (w/w) and the oral administration time as a course of treatment was 20 days.

Angelica sinensis↗

Production of swainsonine by fungal endophytes of locoweed.

Consumption of locoweeds, legumes endemic in arid western USA, has long been associated with locoism, a disease of ruminant animals. To explore the relationship between fungi associated with locoweed and locoweed toxicity, 11 locoweed populations from various sites in New Mexico were assessed for endophytic fungi. Endophytes were isolated from the leaves, stems, seeds, and flowers of eight populations of the toxic locoweeds Astragalus mollissimus, Oxytropis lambertii, and O. sericea. Fungal cultures grew very slowly and sporadically produced subcylindrical conidia with very dark transverse septa. All cultured endophytes produced the alkaloid swainsonine, which causes locoism. Endophyte-infected locoweed populations produced swainsonine, and the swainsonine level of endophyte strains in vitro was highly correlated with the swainsonine level of their host plant populations. The rDNA ITS from mycelia from four endophyte isolates and beta-tubulin encoding regions from mycelia of 18 fungal endophyte isolates were amplified using PCR and the nucleic acid sequences were analyzed. The nucleic acid sequences of the beta-tubulin encoding regions were essentially identical among all the endophytes regardless of plant genus and locations. Morphological evidence and sequence analysis of the ITS region suggest that the endophytes are most closely related to Embellisia. However, with the paucity of Embellisia species represented in sequence databases, precise taxonomic placement will await further study.

Animals↗

Biomedical applications of poisonous plant research.

Research designed to isolate and identify the bioactive compounds responsible for the toxicity of plants to livestock that graze them has been extremely successful. The knowledge gained has been used to design management techniques to prevent economic losses, predict potential outbreaks of poisoning, and treat affected animals. The availability of these compounds in pure form has now provided scientists with tools to develop animal models for human diseases, study modes of action at the molecular level, and apply such knowledge to the development of potential drug candidates for the treatment of a number of genetic and infectious conditions. These advances are illustrated by specific examples of biomedical applications of the toxins of Veratrum californicum (western false hellebore), Lupinus species (lupines), and Astragalus and Oxytropis species (locoweeds).

Abnormalities, Drug-Induced↗

Application of 3' untranslated region (UTR) sequence-based amplified polymorphism analysis in the rapid authentication of Radix astragali.

Radix astragali (root of Astragalus membranaceus) is an important traditional Chinese medicine. It has been used as a tonic herb for thousands of years in China. The water extract of the roots has a wide range of immunopotentiating effects and has been proven to be efficacious as an adjunct cancer therapy. Authentication of the herbal plant is routinely required for general practice in the field of herbal medicine. To facilitate rapid identification of numerous varieties of Radix astragali that are circulating on the herb markets, a rapid molecular genetic method, named 3' untranslated region (3' UTR) sequence-based amplified polymorphism (UAP), has been developed. A cDNA library was first built from transcripts of an authentic A. membranaceus species. Several cDNA clones specific to A. membranaceus were identified through subtractive hybridization of the A. membranaceus cDNA library with Arabidopsis total cellular RNA. On the basis of these cDNA sequences of the 3' untranslated region (3' UTR) of selected cDNA clones, a Polymerase Chain Reaction (PCR) was performed on genomic DNAs of the dry roots of several putative A. membranaceus. PCR fragment length polymorphism was found between A. membranaceus and its relatives. By using this method, it was possible to differentiate the authentic A. membranaceus root from those putative ones obtained from herbal medicine markets. To the authors' knowledge, this is the first paper applying UAP in the authentication of traditional Chinese medicine plants.

3' Untranslated Regions↗

Suppression of transforming growth factor-beta1 gene expression by Danggui buxue tang, a traditional Chinese herbal preparation, in retarding the progress of renal damage in streptozotocin-induced diabetic rats.

Danggui buxue tang (DBT), a preparation containing Angelica sinensis (danggui) and Astragalus membranaceus (huangqi) at a ratio of 1 : 5, is used widely in China for stimulating red blood cell production and enhancing cardiovascular function. The present study was undertaken to characterize the effects of this preparation on diabetic nephropathy using streptozotocin-diabetic rats as a model. Streptozotocin-dependent alterations in renal weight/body weight ratio, urinary albumin and beta (2)-microglobulin concentrations, urinary albumin excretion rate, and creatinine clearance were ameliorated after eight weeks of treatment with either DBT or the angiotensin-converting enzyme inhibitor, benazepril. DBT, but not benazepril, partially attenuated the increases in blood glucose, triglycerides and cholesterol in STZ-diabetic rats. Additionally, the increased expression of transforming growth factor-beta (1) mRNA in the renal cortex due to streptozotocin-induced diabetes was modestly attenuated by these treatments. However, eight weeks of treatment with DBT failed to modify the concentration of angiotensin II in plasma or kidney, indicating that the ability of the preparation to retard the progression of kidney disease was not attributable to inhibition of the renin-angiotensin system. We propose that DBT alleviates renal alterations in diabetes and slows the progression of diabetic nephropathy by suppressing transforming growth factor-beta (1) mRNA expression. The preparation may therefore be useful as an adjuvant therapy for controlling diabetes and its complications.

Angelica sinensis↗

Effects of mushroom and herb polysaccharides, as alternatives for an antibiotic, on growth performance of broilers.

(1) This in vivo trial was conducted to study the effects of polysaccharide extracts of two mushrooms, Lentinus edodes (LenE) and Tremella fuciformis (TreE), and a herb, Astragalus membranaceus (AstE) on growth performance, and the weights of organs and the gastrointestinal tract (GIT) of broiler chickens. (2) Three extracts (LenE, TreE and AstE) were supplemented at inclusion rates of 0.5, 1, 2, 3 and 4 g/kg from 7 to 14 d of age and compared with an antibiotic treatment group (20 mg/kg, virginiamycin (VRG) as well as a group of non-supplemented birds. (3) Body weight (BW) gain, feed intake and feed conversion ratio (FCR) of the extract-supplemented groups were not significantly different from those of the antibiotic group. Significant effects of type of extract and concentration on growth performance were found from 7 to 28 d of age. Generally, birds fed with LenE showed higher BW gain and lower FCR from 7 to 28 d of age than those fed with TreE and AstE and 2 g/kg LenE was considered the optimal inclusion rate for enhanced broiler growth. However, the extracts had no significant effect on the relative weights of organs and GIT compared with the antibiotic group. (4) The birds fed the extracts showed better growth performance than the non-supplemented birds, but were not significantly different from those fed VRG. Of the three extracts, LenE appeared to be a potential growth promoter. Future studies are needed to investigate whether the extracts can be used as alternatives for antibiotic growth promoters in challenged birds, and to elucidate the mechanisms for potentially enhanced growth performance in poultry.

Animal Nutritional Physiological Phenomena↗

Empirical models of pollen limitation, resource acquisition, and mast seeding by a bee-pollinated wildflower.

Synchronous mast seeding is increasingly recognized as common in plant populations. Recent theoretical models show that synchronous mast seeding could be a consequence of resource allocation and storage within individual plants, coupled by pollen limitation in low-flowering years. We used long-term population and weather data to parameterize models of flowering based on stored resources and pollen limitation in Astragalus scaphoides, a bee-pollinated plant that flowers in alternate years. We used these models to test whether internal resource dynamics could explain mast seeding in A. scaphoides and, if so, whether synchrony was caused by pollen limitation and/or fluctuations in precipitation. We compared predictions of models that included all combinations of three factors: constant versus precipitation-dependent resource gain, uniform versus heterogeneous resource gain (among individual plants), and resource-dependent versus resource- and pollen-limited fruit set. Pollen limitation and heterogeneous resource gain were necessary and sufficient to explain alternate-year flowering, but precipitation increased the quantitative match between model predictions and flowering dynamics. Together, our results support the importance of density-dependent pollen limitation as an ultimate and proximate cause of mast seeding in A. scaphoides. Precipitation does not act as a direct cue for synchrony in this species but might affect long-term resource gain and fruiting dynamics.

Animals↗

Chinese medicinal herbs inhibit growth of murine renal cell carcinoma.

Tumors are known to produce factors suppressing immune functions. We previously showed that a murine renal cell carcinoma (Renca) suppressed macrophage function in vitro and that this suppression was abolished by co-incubation with extracts of two Chinese medicinal herbs. We now report that these phytochemicals are capable of inhibiting growth of Renca in vivo. BALB/c mice were transplanted intraperitoneally (IP) with 1-2 x 10(5) Renca cells. One day after tumor transplant, mice were randomized into two groups. One group was treated IP, daily for 10 days, with 100 microliters of phytochemicals containing 500 micrograms each of Astragalus membranaceus and Ligustrum lucidum, while the other group received saline as controls. A cure rate of 57% was obtained with these phytochemicals when the initial tumor load was 2 x 10(5), and 100% when the initial tumor load was 1 x 10(5). Additional experiments were performed to investigate the mechanisms involved in this protection. Splenic macrophages from tumor-bearing mice were shown to have depressed chemiluminescent oxidative burst activity, and this depression was restored with phytochemical treatment. Splenocytes from mice transplanted with Renca responded less favorably to interleukin-2 (IL-2) in generating lymphokine-activated killer (LAK) cells; again this depression was restored with phytochemical treatment. Our data suggest that these phytochemicals may have exerted their antitumor effects via augmentation of phagocyte and LAK cell activities.

Animals↗

In vitro fermentation characteristics of two mushroom species, an herb, and their polysaccharide fractions, using chicken cecal contents as inoculum.

In vitro fermentabilities of two mushrooms (Lentinus edodes--LenS; Tremella fuciformis--TreS), an herb (Astragalus membranaceus--AstS), and their polysaccharide fractions (LenE, TreE, and AstE) were investigated using microflora from chicken ceca. Polysaccharides were extracted using the hot water method. The mushrooms had lower polysaccharide yields (8 to 10%) than the herb (31%). Fermentation kinetics were determined using the in vitro cumulative gas production technique. End-products, such as gas, volatile fatty acids (VFA), and ammonia, were also determined. The gas profiles of intact materials were similar for AstS and LenS. The TreS had a diphasic digestion pattern. The extracts had similar profiles to the intact materials though gas production rates were faster. Intact materials tended to produce less VFA than the extracts though LenS and AstE had the highest total VFA production overall. Intact materials contained more protein than the extracts, and therefore resulted in more branched-chain fatty acids and ammonia. Fermentation kinetics and end-point products demonstrated differences in availability of substrates between the mushrooms and herb. These medicinal mushroom and herb materials, particularly their polysaccharide extracts, show promise in altering microbial activities and composition in chicken ceca. In vivo experiments are necessary for confirmation of this hypothesis.

Ammonia↗

Effects of mushroom and herb polysaccharides on cellular and humoral immune responses of Eimeria tenella-infected chickens.

We investigated the effects of polysaccharide extracts from 2 mushrooms, Lentinus edodes (LenE) and Tremella fuciformis (TreE), and an herb, Astragalus membranaceus (AstE), on cellular and humoral immune responses of Eimeria tenella-infected chickens. A total of 150 broiler chicks were assigned to 5 treatment groups: 3 groups were infected with E. tenella and fed with extracts (LenE, TreE, and AstE), and 2 control groups were infected with or without E. tenella. The 3 extracts were given at the level of 1 g/kg of the diet from 8 to 14 d of age. Specific systemic and cecum mucosal antibody production, proliferation of splenocytes, and peripheral T and B lymphocytes were measured during the 3 wk following Eimeria infection. A significantly higher production of specific IgA, IgM (at d 14 and 21 postinfection), and IgG (at d 21 postinfection) were detected in the Eimeria-infected groups fed with the extracts than in the infected group not fed with the extracts. Of the 3 extracts, TreE stimulated a slightly higher production of specific IgM (P = 0.052), and a significantly higher IgG production at 21 d postinfection. The cecal antibody production showed a similar trend to that of serum antibodies. The overall mean levels of cecal-specific IgA and IgG of the groups fed with extracts were significantly higher at 14 and 21 d postinfection compared with the group not fed with extracts. Of the 3 extracts, the AstE-fed group showed the highest IgG production at d 7 postinfection. Both TreE- and LenE-fed groups had significantly higher IgM and IgG levels compared with the AstE group at d 21 postinfection. The extract-fed groups also showed a significantly higher antigen-specific proliferation of splenocytes at 14 and 21 d postinfection compared with the group not fed with the extracts. The overall mean of erythrocyte rosette-forming cells (ERFC %) (at d 14 and 21) and erythrocyte-antibody-complement cells (EAC %) (at d 14) of the groups fed with the extracts was significantly higher compared with the group not fed the extracts. It is concluded from this study that supplementation with mushroom and herb extracts resulted in enhancement of both cellular and humoral immune responses in E. tenella-infected chickens.

Animals↗

Imaging of selenium in plants using tapered metal monocapillary optics.

Tapered metal monocapillary optics provide a potential alternative to conventional methods of producing small X-ray beams. This paper presents the initial results of chemically specific imaging using such devices. Cellular resolution of organic selenium is obtained in a longitudinal section of mature Astragalus bisulcatus, a selenium hyperaccumulating plant. This work demonstrates the utility of metal monocapillary optics for imaging dilute levels of target elements in biological tissues.

Astragalus Plant↗

Agrobacterium bioassay strain for ultrasensitive detection of N-acylhomoserine lactone-type quorum-sensing molecules: detection of autoinducers in Mesorhizobium huakuii.

An ultrasensitive bioassay system for the detection of N-acylhomoserine lactones (AHLs) was constructed in Agrobacterium tumefaciens by using the T7 expression system to overproduce the AHL receptor TraR. This strain detected many diverse AHLs, some at extremely low concentrations. We used this strain to detect for the first time AHLs made by Mesorhizobium huakuii, which symbiotically fixes nitrogen in association with the legume Astragalus sinicus, a source of green manure throughout eastern Asia.

4-Butyrolactone↗

Protective effect of astragalosides on myocardial injury by isoproterenol in SD rats.

We have extracted and roughly purified astragalosides (AS) from Astragalus membranaceus, a natural herb used as a traditional Chinese medicine, regarded to have pharmacodynamic benefits of protecting injured myocardium. We hypothesized that the astragalosides might exert beneficial effect in myocardial lesion by preserving both energy metabolism and Ca(2+) homeostasis. Sprague-Dauley (SD) rats were injected with isoproterenol (ISO) subcutaneous (s.c.) at a dose of 5 mg/kg/day consecutively for two days as models and were treated with astragalosides and trimetazidine intraperitoneally (i.p.) respectively, at a dose of 5 mg/kg/day one day prior to isoproterenol for 8 days. The histological changes were alleviated in isoproterenol-injected SD rats treated with astragalosides. Compared with isoproterenol-injected rats, the concentration of myocardial intracellular [Ca(2+)]i was decreased, L-type Ca(2+) current density and sarcoplasmic reticulum (SR) Ca(2+) load were recovered, the concentration of myocardial ATP was increased and phosphocreatine (PCr) was decreased in rats treated with astragalosides. In conclusion, the efficacious treatment of astragalosides for myocardial injury might be through regulating intracellular Ca(2+) homeostasis and energy metabolism.

Adenosine Triphosphate↗

The protective effects of traditional Chinese medicine prescription, han-dan-gan-le, on CCl4-induced liver fibrosis in rats.

Han-Dan-Gan-Le, a Chinese medicine preparation composed of Salvia miltorrhiza, Radix paeoniae, Astragalus membranaceus, Stephania tetrandra, and dried leaves of Ginkgo biloba, has been used successfully to treat human liver fibrosis and cirrhosis for years. This study was designed to examine the mechanisms of the protection. Male Wistar rats were given CCl4 (1.2 ml/kg, 2 times/week), 20% fat diet, and 30% alcohol in drinking water (every other day) for 6 weeks. Han-Dan-Gan-Le (0.5 and 1.0 g/kg, p.o., daily for 6 weeks) was administered to rats simultaneously to examine the protective effects against CCl4-induced liver fibrosis. The experimentally-induced liver fibrosis and other morphological alterations were significantly ameliorated by Han-Dan-Gan-Le. Han-Dan-Gan-Le treatments decreased CCl4-induced hepatic collagen accumulation by more than 50%, and significantly increased urinary excretion of hydroxyproline. The CCl4-induced lipid peroxidation in liver and serum was ameliorated as a result of Han-Dan-Gan-Le treatment, possibly by restoring the activity of superoxide dismutase activity in liver and erythrocytes, In conclusion, Han-Dan-Gan-Le is effective in protecting against liver fibrosis. The mechanisms of the protection appear to be due to its antioxidant properties and the modulation of hepatic collagen metabolism.

Animals↗

Regulation of the fibrinolytic potential of cultured human umbilical vein endothelial cells: astragaloside IV downregulates plasminogen activator inhibitor-1 and upregulates tissue-type plasminogen activator expression.

We have investigated whether the saponin astragaloside IV (AS-IV), a 3-O-beta-D-xylopyranosyl-6-O-beta-D-glucopyranosylcycloastragenol, purified from the Chinese herb drug Astragalus membranaceus, which is used in traditional Chinese medicine to treat cardiovascular diseases, might affect the fibrinolytic potential of cultured human umbilical vein endothelial cells (HUVECs). When HUVECs were conditioned with AS-IV, a dose (0.01-100 microg AS-IV/ml)- and time-dependent decrease in plasminogen activator inhibitor type 1 (PAI-1) and an increase in tissue-type plasminogen activator (t-PA) synthesis were observed, which were significant from 1 microg AS-IV/ml and from 12 h of incubation with 100 microg AS-IV/ml. PAI-1 antigen decreased from 641 +/- 86 to 318 +/- 18 ng/10(5) cells/24 h, whereas t-PA antigen increased from 4.1 +/- 0.3 to 9.7 +/- 0.4 ng/10(5) cells/24 h after addition of 100 microg AS-IV/ml. PAI-1 activity decreased to 30% of control level, whereas t-PA activity and t-PA-PAI-1 complexes reached a maximum stimulation of 3- and 5-fold over control levels, respectively, in the conditioned media of HUVECs treated with 100 microg AS-IV/ml for 24 h. PAI-1-specific mRNA expression decreased to 55% (2.2 kb) and 72% (3.2 kb), 66% (2.2 kb) and 88% (3.2 kb), and 19% (2.2 kb) and 41% (3.2 kb) of control values after incubation for 6, 12 and 18 h, respectively, whereas t-PA-specific mRNA increased 2-, 2.5- and 1.4-fold in HUVECs treated with 100 microg/ml AS-IV for 6, 12, and 18 h, respectively. In conclusion our data give evidence that in fact AS-IV can increase the fibrinolytic potential of cultured HUVECs not only by upregulating the expression of t-PA as NG-R1 does, but also by downregulating the expression of PAI-1.

Antihypertensive Agents↗

Production of Se-methylselenocysteine in transgenic plants expressing selenocysteine methyltransferase.

BACKGROUND: It has become increasingly evident that dietary Se plays a significant role in reducing the incidence of lung, colorectal and prostate cancer in humans. Different forms of Se vary in their chemopreventative efficacy, with Se-methylselenocysteine being one of the most potent. Interestingly, the Se accumulating plant Astragalus bisulcatus (Two-grooved poison vetch) contains up to 0.6% of its shoot dry weight as Se-methylselenocysteine. The ability of this Se accumulator to biosynthesize Se-methylselenocysteine provides a critical metabolic shunt that prevents selenocysteine and selenomethionine from entering the protein biosynthetic machinery. Such a metabolic shunt has been proposed to be vital for Se tolerance in A. bisulcatus. Utilization of this mechanism in other plants may provide a possible avenue for the genetic engineering of Se tolerance in plants ideally suited for the phytoremediation of Se contaminated land. Here, we describe the overexpression of a selenocysteine methyltransferase from A. bisulcatus to engineer Se-methylselenocysteine metabolism in the Se non-accumulator Arabidopsis thaliana (Thale cress). RESULTS: By over producing the A. bisulcatus enzyme selenocysteine methyltransferase in A. thaliana, we have introduced a novel biosynthetic ability that allows the non-accumulator to accumulate Se-methylselenocysteine and gamma-glutamylmethylselenocysteine in shoots. The biosynthesis of Se-methylselenocysteine in A. thaliana also confers significantly increased selenite tolerance and foliar Se accumulation. CONCLUSION: These results demonstrate the feasibility of developing transgenic plant-based production of Se-methylselenocysteine, as well as bioengineering selenite resistance in plants. Selenite resistance is the first step in engineering plants that are resistant to selenate, the predominant form of Se in the environment.

Arabidopsis↗

Gastroprotective effect of Astragaloside IV: role of prostaglandins, sulfhydryls and nitric oxide.

This investigation evaluated the gastroprotective activity of Astragaloside IV, a cycloartane-type triterpene glycoside isolated from Astragalus zahlbruckneri. Gastric mucosal damage was induced in rats by intragastric ethanol (1 mL/rat). Rats treated orally with Astragaloside IV suspended in Tween 80 at 3, 10 and 30 mg kg(-1), showed 15, 37 and 52% gastroprotection, respectively. The gastroprotection observed at 30 mg kg(-1) for this compound was attenuated in rats pretreated with N(G)-nitro-L arginine methyl ester (70 mg kg(-1), i.p), a nitric oxide (NO)-synthase inhibitor, suggesting that the gastroprotective mechanism of this glycoside involves, at least in part, the participation of NO. The gastroprotective effect of Astragaloside IV was not affected by the inhibition of prostaglandin synthesis with indometacin (10 mg kg(-1), s.c.) nor by the block of endogenous sulfhydryls with N-ethylmaleimide (NEM, 10 mg kg(-1), s.c.). Carbenoxolone was used as a gastroprotective model drug and showed a dose-dependent gastroprotective effect (25, 43 and 88% of gastroprotection, at 3, 10 and 30 mg kg(-1), respectively). The partial participation of prostaglandins, sulfhydryls and NO was observed in the gastroprotective mechanism of carbenoxolone.

Animals↗