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Formosanin C (FC) is a pure compound isolated from Rhizoma Paridis. In the past years, antitumor effects of FC have been observed in several cultural cells and animal systems. However, there was no research particular on liver cancer. In this experiment, 3-(4, 5-dimethylthiazol diphenyltetrazolium bromide (MTT) dye reduction assay was used to evaluate cell viability of HepG2 cells with FC-treatment. 4', 6-diamidino-2-phenylindole (DAPI) staining, Annexin V-FITC/PI assay and DNA fragment assay were applied to observe FC-induced apoptosis. Cell cycle analysis and NMR metabolic profiles were used to identify molecular mechanisms of FC in HepG2 cells. As a result, FC inhibited the growth of HepG2 cells through inducing apoptosis and S phase arrest. Cells cultured in the presence or absence of FC was different in metabolic profiles. The treatment decreased acetate, ethanol, choline and betaine, and increased butyrate, fatty acids, leucine and valine in HepG2 cells. In conclusion, metabolomic analysis of the exometabolome of FC-treated HepG2 cells, together with traditional methods such as apoptosis test and cell cycle analysis provided a holistic method for elucidating mechanisms of potential anti-cancer drug, FC.

Ethnopharmacological relevance Wei-Chang-An pill (WCA pill), a traditional Chinese medicine, has been used for treating various gastrointestinal diseases for several decades. Despite the popular medicinal use of WCA pill, less data was available to its activity and mechanism in gastrointestinal disorders. To examine the effects of the methanol extract of WCA pill (ME) on gastrointestinal tract so as to assess some of the possible mechanisms involved in the clinical treatment. Materials and methods ME was studied on gastrointestinal transit in vivo including gastric emptying and small intestinal motility in normal and neostigmine-induced mice, as well as on the isolated tissue preparations of rat jejunum in vitro . Results In vivo , the gastric emptying decreased and intestinal transit increased after administration of ME in normal mice. However, administration of ME accelerated the intestinal transit ranging from 0.01 to 0.8 mg/mL and reduced it at the concentration of 1.6 and 3.2 mg/mL, while the gastric emptying was inhibited throughout the concentrations in neostigmine-induced mice. in vitro , ME caused inhibitory effect on the spontaneous contraction of rat-isolated jejunum in dose-dependent manner ranging from 0.01 to 6 mg/mL and also relaxed the acetylcholine chloride (Ach, 10 −6 M)-induced and K + (60 mM)-induced contractions. ME shifted the Ca 2+ concentration–response curves to right, similar to that caused by verapamil (0.025 mM). Conclusions These results indicated that ME might play a bidirectional role in gastrointestinal transit modulation and the effects on isolated tissue are probably mediated through calcium influx and muscarinic receptors, which provides pharmacological basis for the clinical use of WCA pill in gastrointestinal tract disorders.

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease that causes deformity of the joints and physical disability. Icariin, a natural flavonoid glucoside isolated from plants in theEpimediumfamily, has been proven to have various pharmacological activities. A recent study showed that icariin suppressed cartilage and bone degradation in mice of collagen-induced arthritis. However, the mechanism needs to be further investigated. In our current study, we found that icariin reduced the arthritis score and the incidence of arthritis compared with that in mice treated with water. Icariin inhibits the expression of various osteoclastogenic markers, such asβ3 integrin, cathepsin K, and MMP9in vitro. Icariin treatment in mice with CIA also resulted in less number of Th17 cells and decreased ratio of CD4+IL-17+cells. The alleviated arthritis score and incidence of arthritis and reduced serum levels of IgG2a induced by icariin were abolished with additional IL-17 administration. Furthermore, icariin inhibited STAT3 activation in T cells and STAT3 inhibitor resulted in decreased IL-17 production and alleviated RA. In conclusion, icariin decreases Th17 cells and suppresses the production of IL-17, which contributes to the alleviated rheumatoid arthritis, through the inhibition of STAT3 activation.

Panax quinquefolium L. and Glycyrrhiza uralensis Fisch. are important medicinal plants and health food that are used worldwide. Field cultivation of Panax quinquefolium and Glycyrrhiza uralensis is an extremely time consuming and labor-intensive process. Plant cell culture offers an alternative for obtaining valuable chemicals, especially plant-specific bioactive secondary metabolites. In this review, cell suspension cultures of Panax quinquefolium and Glycyrrhiza uralensis are described for the production of bioactive compounds.

The aim of this paper is to research the protective effect of Astragalus against intermittent hypoxia-induced hippocampal neurons impairment in rat and lay the theoretical foundation for sleep apnea improvement in cognitive function by Astragalus. Male Wistar rats were divided into four groups: (1) blank control group; (2) normoxia group; (3) intermittent hypoxia group; (4) Astragalus treated intermittent hypoxia group. After 6-week treatment, the expression of iNOS was detected by real-time reverse transcription polymerase chain reaction (RT-PCR) at mRNA level as well as by immunohistochemistry (IHC) and western blot at protein level. As a result, Astragalus reduced the expression of iNOS at mRNA and protein levels in hippocampus compared with non-treated groups (P < 0.05). In conclusion, Astragalus could protect intermittent hypoxia-induced hippocampal neurons impairment in rat.

Weichang’an (WCA) pill, a traditional Chinese patent medicine consisting of ten Chinese medicinal herbs, has been used to treat irritable bowel syndrome and functional dyspepsia for several decades. In this study, twelve bioactive constituents in the methanol extract of WCA were accurately identified since MS/MS fragmentation behavior of the references and the standards by using HPLC-DAD-ESI-MS/MS analysis and a reliable and accurate method for the simultaneous determination was developed. Twelve active components including costunolide and dehydrodehydrocostus lactone from the principal herb Radix Aucklandiae; naringin, hesperidin and neohesperidin from Fructus Aurantii; magnolol and honokiol from the ministerial herbs Cortex Magnoliae officinalis; aloe-emodin, rhein, emodin, chrysophanol and physcion from adjunctive and messenger herb Radix et Rhizoma Rhei were analyzed in the samples. The chromatographic separation was performed on a Kromasil C18 column with gradient elution of acetonitrile-methanol and 1.0% acetic acid water. In this condition, linearity, inter- and intra-day precision and accuracy were within acceptable ranges. The developed method showed satisfactory precision and accuracy with overall intra- and inter-day variations of 0.68-1.33% and 0.67-2.05% respectively, and the overall recoveries of 97.54-102.69% for twelve compounds. The proposed approach was successfully applied as a powerful tool for the quality control of WCA pill.

Cerebralcare Granule (CG), one of the famous classical recipes in traditional Chinese medicine, is developed from the "Decoction of Four Drugs". It has been used for treatment of cerebrovascular related diseases, such as hypertension. It is well known that vasodilatation plays a very important role in hypertensive. Despite the popular medicinal use of CG, little data was available to its activity and mechanism involved in vasodilatation. Therefore, we aimed to investigate the vasorelaxant effects of CG on isolated rat thoracic aorta so as to assess some of the possible mechanisms. The present study was performed to examine the vasodilative activity of CG and its mechanisms in isolated rat thoracic aorta.CG was studied on isolated rat thoracic aorta in vitro, including endothelium-intact and endothelium-denuded aortic rings. In present study, specific inhibitors including NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME), cyclooxygenase (COX) inhibitor indomethacin (INDO), non-selective K+ channel inhibitor tetraethylammonium chloride (TEA), Kir channel inhibitor BaCl2, KATP channel inhibitor Glibenclamide (Gli) and cholinergic receptor antagonist atropine were used, they were added 20 min before NE contraction and then added CG-induced vasodilation.Removal of endothelium or pretreatment of aortic rings (intact endothelium) with L-NAME (0.1 mM) or INDO (0.01 mM) significantly blocked the CG induced relaxation. Pretreatment with the non-selective K+ channel inhibitor TEA (1 mM), or the Kir channel inhibitor BaCl2 (0.1 mM), neither of them had no influence on the CG-induced response (p0.05). However, pretreatment with the KATP channel inhibitor Gli (0.01 mM) produced significant inhibition on the CG-induced response (p0.01). Besides, CG also inhibited the contraction triggered by NE in endothelium-denuded rings in Ca2+-free medium. CG (0.4, 0.8 and 3.2 mg/mL) produced rightward parallel displacement of CaCl2 curves and reduced the maximum contraction induced by 30 mM CaCl2 to 31.1±9.3%, 18.8±6.9% and 9.4±4.5%, respectively. The relaxation, induced by CG on endothelium-intact rat aortic rings pre-contracted with NE, was significantly attenuated in the presence of atropine (EC50=3.7 mg/mL, p0.01).Our results suggest that CG induces relaxation in rat aortic rings through an endothelium-dependent pathway mediated by NO/cGMP pathway and an endothelium-independent pathway involving blockade of Ca2+ channels, inhibition of Ca2+ mobilization from intracellular stores, opening of KATP channel. In addition, the muscarinic receptor stimulation is also one of the vasorelaxant mechanisms.

The ultrastructure of the endolymphatic sac (ES) of the late stage larva of the Japanese red-bellied newt, Cynops pyrrhogaster (stage 57), was examined by light and transmission electron microscopy. The two endolymphatic sacs are located at the dorsal-medial side of the otic vesicle on the dorsal-lateral side of the midbrain in the cranial cavity. The wall of the sac is composed of a layer of cubical epithelial cells with loose, interposed intercellular spaces. The sac contains a large luminal cavity, in which endolymph and numerous otoconia are present. The epithelial cells of different portions of the sac have a similar structure. These cells contain an abundance of cytoplasmic organelles, including ribosomes, Golgi complexes, and numerous vesicles. Two types of vesicles are found in the epithelial cells: the "floccular" vesicle and the "granular" vesicle. The floccular vesicles are located in the supra- and lateral-nuclear cytoplasm and contain floccular material. The granular vesicles have a fine granular substance and are usually situated apposed to the apical cell membrane. The granular vesicles are suggested to be secreted into the lumen, while the floccular vesicles are thought to be absorbed from the lumen and conveyed to the intercellular spaces by the epithelial cells. The apical surfaces of the epithelial cells bear numerous microvilli. Apparently floating cells, which bear long microvilli on the free surfaces, are observed in the lumen of the ES. Based on the fine structure, the function of the endolymphatic sac of the newt Cynops pyrrhogaster is discussed.

The development of the statocyst of the freshwater snail Biomphalaria glabrata has been examined from embryo to adult. Special emphasis was put on the growth of the statoconia in the statocysts. In the statocysts of embryonic snails (90-120 h after oviposition) there is not a single statolith but an average of 40-50 statoconia per statocyst. The number of statoconia increases to 385-400 when the snails reach a shell diameter of 4 mm and remains relatively constant thereafter, irrespective of shell size. Small statoconia are found in supporting cells, which suggests that the statoconia are produced within these cells. The average diameter of statoconia and the total mass of statoconia increase with increasing shell diameter. The average number of large statoconia (diameter greater than 7 micrometers) per statocyst continues to increase from 2 to 10 mm animals while the number of small ones (diameter less than 4 micrometers) initially rises and then decreases after 4 mm. These results demonstrate continuous growth of the statoconia in the cyst lumen of Biomphalaria. The single statoconia vibrate in a regular pattern in vivo, indicating beating of the statocyst cilia. The statoconia sink under the influence of gravity to load and stimulate receptor cells which are at the bottom. The length of cilia and the size of statocyst gradually increase as the animal grows. However, the increase in the volume of the statocyst is relatively small compared with the increase in body weight during normal development.

To find out a simpler method that can directly transfer the aim gene into plant genomes, the purple medic seeds irradiated by 60 Co with 0.375 Gy were transformed by linear DNA containing a β-glucuronidase (GUS) gene (as an aim gene), a betaine aldehyde dehydrogenase (BADH) gene (as a selectable marker) and two pairs of both CaMV35S promoter and Nos terminator. Subsequently, the seeds were planted and grown in perlite media watered with NaCl solution as a kind of selective compound. The results showed that, positive frequency of PCR identification by the GUS gene or the BADH gene was higher than 53.2 and 89.5% in T 0 and T 1 generations, while GUS staining rate was higher than 50%; whereas five T 1 plants assayed by southern hybridization all showed positive reaction. In conclusion, by this method, transgenic plants may be easily obtained with the antibiotic markers for free; moreover, the plant regeneration-system must not be erected by directly transforming the seeds. Key words : DNA transformation, irradiated seeds, purple medic, salt screening.

Rhizoma Paridis Saponins (RPS), which is the effective part of Rhizoma Paridis, showed strong anti-lung cancer and anti-hepatocarcinoma activities. In this research, a gas chromatography/mass spectrometry (GC/MS) method was developed and validated for the metabolic profiling of RPS intervention in H22 hepatocarcinoma mice. Data were analyzed with partial least-squares discrimination analysis (PLS-DA). As a result, RPS displayed different pathway to decrease energy production of the mice. For the normal mice, RPS significantly decreased the concentration of lipid, glycerate, succinate and lactate, but increased glucose and valine levels. All these indicated that RPS inhibited glucose and valine to transform ketones which participated in the ATP production. For the H22 cancer mice, RPS increased the concentration of lipid and glycerate, but significantly decreased glucose, glycine and alanine levels in the serum. This phenomenon indicated that RPS inhibited the oxidation of fatty acids pathway and the gluconeogenesis pathway which participated in the energy supply for the body. RPS also inhibited glycine and alanine production to block the tumor growth. This selective effect of RPS to different condition of mice would improve understanding of the antitumor pathway of RPS involved in H22 hepatocarcinoma mice.

Ethnopharmacological relevance Radix Aucklandiae (RA), a well known traditional Chinese medicine, is widely used for treating various problems in digestive system. A selective and sensitive high-performance liquid chromatography coupled with mass spectrometry method was first developed and validated for simultaneous quantification of costunolide and dehydrocostuslactone in rat plasma with diazepam as internal standard after oral administration of RA extraction. Materials and methods Plasma samples were extracted via solid-phase extraction and detected by multiple-reaction monitoring mode under positive electrospray. Chromatographic separation was accomplished on an Agilent C18 column (2.1 mm×150 mm, 5 µm), with 0.1% formic acid and acetonitrile (1:1) as the mobile phase at a flow rate of 0.5 mL/min. Results The quantification was performed using the transitions of m/z 233/187 for costunolide, m/z 231/185 for dehydrocostuslactone and m/z 285/193 for diazepam, respectively. Calibration curves were linear over the concentration range of 0.7–769.7 ng/mL for costunolide and 0.9–956.0 ng/mL for dehydrocostuslactone. The intra-day and inter-day precisions (RSD%) for two compounds was less than 8.76% and 9.70% and the accuracy (RE%) range from 6.14% to 5.35%. The time to reach the maximum plasma concentration (Tmax) was 10.46 h for costunolide, 12.39 h dehydrocostuslactone. The elimination half-time (t1/2) of costunolide and dehydrocostuslactone was 5.54±0.81 and 4.32±0.71 (h). The AUC of costunolide and dehydrocostuslactone was 308.83 and 7884.51 respectively (ng h/mL). Conclusions It was the first report for the study of pharmacokinetic profile of costunolide and dehydrocostuslactone in rat plasma after oral administration of RA extract. These results provided a meaningful basis for better understanding the absorption of traditional medicine, RA, and provide useful scientific data for clinical application.

Currently, famous traditional Chinese medicine formulas have undergone re-evaluation and development in China. Wei-Chang-An-Wan (WCAW) as one of them has been used for treating various gastrointestinal diseases for several decades. The secondary development of WCAW is in progress so as to interpret the effective material basis or find new pharmacological activity.To evaluate the antinociceptive effect of methanol extract of WCAW (ME) as well as four fractions (P.E., EtOAc, n-BuOH, H2O) and obtain information on the correlation between the contents of the fractions and antinociceptive effect.ME was divided into four parts extracted by petroleum ether, ethyl acetate and n-butanol. Antinociceptive activity was evaluated by three models of acetic acid-induced writhing, formalin and hot-plate test in mice after repetitive administration of ME at 200, 400 or 800 mg/kg, P.E. 132 mg/kg, EtOAc 106 mg/kg, n-BuOH 176 mg/kg and H2O 176 mg/kg for six days. The chemical compounds were analyzed by HPLC-ESI-MS.ME at 800 mg/kg inhibited acid-induced writhing by 84.69%, and reduced the licking time of second phase in formalin test by 53.23%. The inhibition rates in acid-induced writhing of P.E., EtOAc, n-BuOH and H2O were 27.79, 33.85, 38.97 and 37.69%, respectively, and in formalin test about 50%. They had no effect on the hot-plate test. HPLC-ESI-MS analysis showed that 68 chemical compounds were detected and 41 compounds were identified from ME.The results obtained herein indicate that WCAW possesses the antinociceptive activity that provides a new aspect in clinical application.

With the modernization of traditional Chinese medicine, medicinal plants resources cannot meet the request of Chinese medicine industry. Medicinal plants adventitious roots culture in a large scale is an important way to achieve Chinese medicine industrialization. However, how to establish good adventitious roots culture system is its key, such as plant hormones, explant, sucrose, innoculum and salt strength.

Saponins are active compounds in natural products. Many researchers have tried to find the method for knowing their concentration in herbs. Some methods, such as solid-liquid extraction and solvent extraction, have been developed. However, the extraction methods of the steroidal saponins from Paris polyphylla Smith var. yunnanensis (Liliaceae) are not fully researched.To establish a simple extraction method for the separation of steroidal saponins from the rhizomes of P. polyphylla Smith var. yunnanensis.Macroporous adsorption resins were used for the separation of steroidal saponins. To select the most suitable resins, seven kinds of macroporous resins were selected in this study. The static adsorption and desorption tests on macroporous resins were determined. Also, we optimized the temperature and the ethanol concentration in the extraction method by the contents of five kinds of saponins. Then, we compared the extraction method with two other methods.D101 resin demonstrated the best adsorption and desorption properties for steroidal saponins. Its adsorption data fits best to the Freundlich adsorption model. The contents of steroidal saponins in the product were 4.83-fold increased with recovery yields of 85.47%.The process achieved simple and effective enrichment and separation for steroidal saponins. The method provides a scientific basis for large-scale preparation of steroidal saponins from the Rhizoma Paridis and other plants.

The fermentation processing of traditional Chinese medicines (TCMs), as an important processing method for TCM, originated from the ancient brewing technology. It has a long history in China. Fermented TCMs (FTCMs) are widely applied among folks for preventing and treating many diseases. There are many kinds of TCM processed by spontaneous fermentation, including Massa Medicata Fermentata, Rhizoma Pinelliae Fermentata, Red fermented rice, Semen Sojae Praepaaratum, Mass Galla chinesis et camelliae Fermentata and Pien Tze Huang. This essay summarizes historical origin, main varieties, the effect of microbial strains, current processing techniques and existing problems of FTCM, and look into the prospect of modern development of FTCMs.

OBJECTIVE To compare steroidal saponins-main active ingredients of Paris polyphylla from different areas in Guizhou province, in order to provided basis for further proving Guizhou province to be the planting base of P. polyphylla. METHOD The contents of nine steroidal saponins, namely paris-VII, (25R)-5-en-spirost-3beta,17alpha-diol-3-O-alpha-L-rhamnopyranosyl-(1-->2) [alpha-L-rhamnopyranosyl(1-->4)]-beta-D-glycopyanoside (PGRR), paris-H, paris-VI, paris-II , paris-III, gracillin, paris-I and paris-V of P. polyphylla from eight areas of Guizhou province were determined by HPLC-UV. RESULT Five steroidal saponins-paris-VII, PGRR, paris-H, paris-VI and gracillin were detected in all the drugs, which provided basis for distinguishing P. polyphylla. P. polyphylla from Dushan showed the highest content of steroidal saponins (9.62%), followed by Tongren (6.39%), and the lowest was Zunyi (0.99%). CONCLUSION Saponins from different areas of Guizhou province show significant difference in variety and content, therefore Dushan is suitable for planting P. polyphylla.

Astragali Radix, as a tonic with a long history, is one of the most common traditional Chinese medicines. Recently, chemical composition and pharmacological activities of Astragali Radix have been widely reported. Polysaccharide, triterpenes and flavonoids are regarded to its effective components. Pharmacological experiments and clinical reports show that Astragali Radix has good effects on immune-regulation, anti-tumor, anti-virus, diabetes, anti-aging and so on. Therefore, it shows a good prospect to make good use of its effective components.

Ordered ZnO arrays with a peculiar nanostructure were synthesized by a multistep synthesis process. The first step was the preparation of ZnO seed to induce the formation of ZnO array via potentiostatic electrodeposition method using a typical three electrode set-up. The second step was fabricating ZnO array along seed by Chemical Bath Deposition. Structural analysis of ZnO was carried out with X-ray diffraction (XRD), which showed a hexagonal wurtzite structure, and the selected area electron diffraction (SAED) patterns indicated that nanocrystalline is a part of monocrystal. The scanning electron microscope (SEM) and transmission electron microscope (TEM) were used to study the microstructure, and showed a pagoda-like microstructure with a tiny top and large bottom, which had an average top diameter of exceeding 800[Formula: see text]nm at seed-depositing time of 60[Formula: see text]s, and then growth mechanisms are subsequently given a further explanation, viewed from kinetics and thermodynamics. In addition, the current–voltage curves of schottky diode devices with ZnO nanopagoda arrays revealed that ZnO films arrays along grown ZnO seed had a higher reverse saturation current than ZnO films grown without seed, which are [Formula: see text] A and [Formula: see text] A, respectively. The minimum turn-on voltage of the diode with ZnO seed deposited 60[Formula: see text]s is 0.18[Formula: see text]V, without seed is 0.52[Formula: see text]V.

Xuan-Ju agent is an herbal formula containing aqueous extract of Formica fusca, Herba epimedii, Fructus cnidii, and Fructus lycii, all of which are reputed for their beneficial effects in the treatment of the immunodeficient diseases such as rheumatoid arthritis. We performed a study on the anti-inflammatory effects of this agent using carrageenan- and adjuvant-induced paw edema in rats. Xuan-Ju agent showed a marked inhibitory effect on edema in two models of inflammation in rats, at the dose of 0.20, 0.40 and 0.80 g/kg. Based on this study, Xuan-Ju agent is considered to be a potentially useful drug suitable for further evaluation for rheumatoid arthritis.

Rhizoma paridis steroidal saponins (RPS) have been prepared and identified as the active compounds for antitumor activity in our previous study. However, the low oral bioavailability of the steroidal saponins restricted its using. In the present research, solid dispersion (SD) and phytosome (PHY) formulation of RPS were prepared, and the physicochemical parameters as well as the intestinal absorption in rat everted gut sac model were investigated. Seven agents were selected as the carriers of SD, and poloxamer 407 (P 407) was the most suitable one. SD reduced the particle size of saponins in the water solution, enhanced the solubility of the saponins by about 3.5 folds, and significantly improved the absorption transport of saponins from 48 to 104 μg in everted gut sac of the rat system. PHY significantly enhanced the hydrophilic of saponins but showed little effect on the absorption in small intestine. Jejunum and ileum part absorbed more absolute contents of total saponins than duodenum parts. Six saponins, the main contents of RPS, used as the index of comparing the three forms, were also further investigated in the physico-chemical properties and the absorption tests. n-Octanol/water partition coefficients of the six saponins ordered in RPS, SD and PHY were Chonglouoside H>Dioscin>Polyphyllin D>Gracillin>Paris-VII>Formosanin C. All the saponins possessed the higher absorptive characteristics in SD formulation. The absorption rate of diosgenyl saponins in intestine was more than the pennogenyl saponins.

Objective To optimize different processing techniques of Massa Medicata Fermentata. Method Single factor test was adopted, with the amylase activity of Massa Medicata Fermentata as the assessment indicator, to observe the influence of such factors as fermentation time and mixture techniques of active pharmaceutical ingredients on the amylase activity of Massa Medicata Fermentata. Meanwhile, Massa Medicata Fermentata prepared with the optimum processing techniques and superior and inferior products of Massa Medicata Fermentata in the market were compared in amylase activity, soluble starch content and soluble polysaccharide content. Result The optimum fermentation time was 7 days. Adzuki bean shall be boiled before mixed with other materials. Artemisia annua, Polygonum hydropiper and Cocklebur grass shall be evenly mixed water decoction. The amylase activity, the soluble starch content and the soluble polysaccharide content of fermented Massa Medicata Fermentata could reach to 49.372 mg x min(-1) x g(-1), 7.967%, and 16.65% respectively, significantly higher than the two types of Massa Medicata Fermentata sold in the market. Conclusion According to the optimum processing techniques, Adzuki beans were smashed and mixed equally with flour and Armeniacae Semen Amarum powder, and then successively added with A. annua, P. hydropiper and C. grass for even mixture. The fermentation time was 7 days.

Objective To identify the yeast strains isolated from Massa Medicata Fermentata samples that sold in markets. Method The strains were identified through conventional classification methods including colony characteristics, cell morphology, physiological and biochemical properties, as well as 26S rDNA sequence analysis. Result The isolated strains Y1, Y3, Y4, Y5 were Cryptococcus albidus, Saccharomyces cerevisiae, Pichia kudriavzevii, Endomyces fibuliger, respectively. Conclusion After fermentation the Massa Medicata Fermentata samples contained a variety of yeast species. Yeasts were the main contribution microorganism of the fermentation process.

The changes in the main nutrient and medicinal components during the storage of the Chinese yam (Dioscorea opposita) tubers were studied. The harvested tubers were stored under ambient conditions (10 °C to 18 °C, 60 % to 80 % Relative Humidity) and cold temperature and packaged conditions (4 °C, 60 % to 65 % Relative Humidity) for 45 day. The allantoin, starch, total alcohol-soluble sugar, reducing sugar, protein, and moisture contents of the samples were evaluated. Their amylase activities were also investigated. Results of ambient conditions indicated that, during storage, moisture decreased by 67.96 % to 56.51 %, and total sugars, reducing sugars, and protein increased by 6.49 % to 9.81 %, 1.7 % to 2.27 %, and 13.02 % to 14.55 %, respectively. Starch and enzyme activities increased during the early days of storage and progressively decreased, and the content of allantoin changed in volatility. The changes were more significant at cold temperatures and packaged conditions than at ambient conditions. This result suggests that after-ripening occurred in the early stages of Chinese yam tubers, which positively affected the nutritional potential of the tubers by a marked increase in nutrients. Low-temperature sweetening greatly affects the nutritional potential of tubers by a series of complicated interactions between starch and sugars at 4 °C.

Ethnopharmacological relevance Fructus Aurantii is one of the commonly used traditional Chinese medicines and the compatibility of Fructus Aurantii, Radix et Rhizoma Rhei and Semen Crotonis Pulveratum were the main components in a Chinese traditional patent medicine Weichang'an pill. Naringin and neohesperidin are two main flavonoid glycosides in the extract of Fructus Aurantii. The pharmacokinetics of naringin and neohesperidin, as the main marker constituents were analysis by a simple and accurate method. Materials and Methods In the present study, pharmacokinetic differences of two flavonoid glycosides were detected after oral administration of the extract of single herb and different compatibility of Fructus Aurantii, Radix et Rhizoma Rhei and Semen Crotonis Pulveratum which at doses of approximately 700 mg/kg naringin and 300 mg/kg neohesperidin. Results It indicated that the pharmacokinetic process of naringin and neohesperidin in four groups were demonstrated bimodal phenomenon after oral administration. Comparing to pharmacokinetic parameters of the single herbs administration, there were no significant difference after the compatibility of Fructus Aurantii and Radix et Rhizoma Rhei. However, the absorption of two flavonoid glycosides was increased dramatically after the compatibility of Fructus Aurantii or Semen Crotonis Pulveratum. The same results were attained after co-administration of the extract of three herbs. Conclusion Some components in the extract of Semen Crotonis Pulveratum promoted the absorption of naringin and neohesperidin.

Many anticancer drugs are obtained from natural sources. Nature produces a variety of toxic compounds, which are often used as anticancer drugs. Up to now, there are at least 120 species of poisonous botanicals, animals and minerals, of which more than half have been found to possess significant anticancer properties. In spite of their clinical toxicity, they exhibit pharmacological effects and have been used as important traditional Chinese medicines for the different stages of cancer. The article reviews many structures such as alkaloids of Camptotheca acuminata, Catharanthus roseus and Cephalotaxus fortunei, lignans of Dysosma versipellis and Podophyllum emodi, ketones of Garcinia hanburyi, terpenoids of Mylabris and Ginkgo biloba, diterpenoids of Tripterygium wilfordii, Euphorbia fischeriana, Euphorbia lathyris, Euphorbia kansui, Daphne genkwa, Pseudolarix kaempferi and Brucea javanica, triterpenoids of Melia toosendan, steroids of Periploca sepium, Paris polyphylla and Venenum Bufonis, and arsenic compounds including Arsenicum and Realgar. By comparing their related phytochemistry, toxic effects and the recent advances in understanding the mechanisms of action, this review puts forward some ideals and examples about how to increase antitumour activity and/or reduce the side effects experienced with Chinese medicine.

A simple and effective method of determining the phytate content of a single wheat seed was considered necessary for selecting the materials of moderate phytate during early generations breeding. 23 wheat genotypes (16 winter wheat cultivars and 7 spring wheat cultivars) were firstly used as determining samples of phytate content (PC) and inorganic phosphorus content (IPC) of the whole meal, secondarily they were planted in two locations for two times (2004 and 2005). The all single plants were harvested by a plot from those sites, then 6 individual plants were randomly selected to save their seeds for each plot, and the seeds of other plants were grinded into whole meal to assay their PC and IPC. Subsequently, every seed from per individual plant was cut into two semi-grain seeds: the semi-grain seed without embryo (SGSWOE) and the semi-grain seed with embryo (SGSWE); the SGSWOE was carried out to assay colorimetric scoring values by inorganic phosphorus content (CSVIPC), while according to the CSVIPC index, the satisfied SGSWEs were saved to plant at next generation or the dissatisfied SGSWEs were eliminated. The results showed that there was significantly negative correlation between PC and IPC of the wheat meal from either the first samples or the materials harvested at those sites, and for the latter there was significantly negative correlation between PC and CSVIPC. There was no significant difference of CSVIPC among the SGSWEOs and SGSWE of the spikes and individual plants from the same plot or the same genotype at any sites. In conclusion the CSVIPC of the SGSWOEs can indirectly express PC level of the wheat meal: the higher the former was, the lower the latter was. So the CSVIPC of SGSWOE can be applied to assaying the level of corresponding SGSWE according to their index, and screening the individual plant of moderation phytate. Moreover, this method is characterized by high efficiency during breeding processing.

In this study, seedling, callus, cell, and adventitious root of Glycyrrhiza uralensis Fisch. have been established. In order to find the best one for producing G. uralensis active constituents, triterpenoid saponins and flavonoids in native root and tissue cultures were determined, and the contents in different G. uralensis materials were analyzed using cluster analysis. The contents of triterpenoid saponins and glycyrrhizic acid in tissue cultures were much lower than that in native G. uralensis. The total flavonoids content we determined in adventitious root was 6.31 mg g(-1), which was close to that of native root (9.82 mg g(-1)). Based on the cluster analysis, we found that G. uralensis cultures were not suitable for production of glycyrrhizic acid, while adventitious root had a greater capability of flavonoids production comparing to seedling, callus, and cell.

Objective To study the characteristics of the growth and periplocin accumulation of the adventitious roots of Periploca sepium, and on this basis, study the effect of Ag+ and La3+ elicitors on the growth and periplocin accumulation of the adventitious roots. Method The adventitious roots were sampled every four days, and the dry weight and the contents of the periplocin were measured. The curves of the growth and periplocin accumulation of the roots were plotted. The Ag+ and La3+ elicitors with different concentrations were added to the medium when the adventitious roots grew in the stable phase to study the optimal concentration which was good to synthesize the periplocin. Besides, the optimal concentration of Ag+ and La3+ elicitors was added to the different growth phases to study the effect of the elicitors on the growth and periplocin synthesis of adventitious roots. Result The characteristics of the growth of adventitious roots of P. sepium showed a typical growth S-Curve, which displayed a half-coupling relationship with the metabolism of periplocin. The optimal concentrations of Ag+ and La3+ elicitors were both 0.05 mmol L(-1). Besides, it was the best period for the Ag+ and La3+ elicitors to elicit the synthesis of periplocin when in the terminally exponential phase. Conclusion The growth of adventitious roots and the accumulation of periplocin show a half-coupling relationship. Besides, the concentration and additive time of Ag and La3+ elicitors obviously influences the growth of adventitious roots and synthesis of periplocin.

The starches obtained from two different yam residues, which were treated with alkali(starch-A) or enzyme (starch-E), were studied and compared with yam starch isolated using ordinary method (starch-O) for morphological, crystalline pattern, thermal, and pasting properties. The results revealed that the amylose content of three starches ranged from 19.47 to 22.17%. The granule surfaces of starch-A and starch-E were as smooth as that of starch-O. The crystalline pattern of the three starches was a C-type. The transition temperatures (To, Tp and Tc) varied from 70.11 to 73.64, 79.23 to 81.74, and 84.30 to 86.65 ºC, respectively. The starch-E showed the highest Δ Hgel, followed by the starch-A, while it was lowest for the starch-O. According to the viscosity measurement, starch-O had the lowest pasting temperature, highest peak viscosity and breakdown viscosity, which were contrary to those of starch-E.

OBJECTIVE To analyze the content of periplocin in different part of the Periploca sepium in vitro plantlet and study its dynamic variation during the process of differentiation. METHOD The seeds were generated seedling under aseptic condition, and the cut hypocotyl was induced to form the callus and adventitious buds on the MS culture medium with the hormone of IBA 0.1 mg x L(-1) + BA 1 mg x L(-1). The seedling was cut down when the buds grew up to 3 cm and then the root was cultured in the 1/2 MS culture medium with the hormone of IBA 0.5 mg x L(-1) to form intact plantlet. Different parts of it were collected and the content of periplocin was measured during the process of differentiation. RESULT The contents of periplocin varied widely in different parts during the process of differentiation, with the highest in the roots and then callus, stem and leaf of intact plantlet, stem and leaf of plantlet without root from high to low. CONCLUSION The periplocin of the secondary metabolite is more likely to be produced and accumulated in root and callus. Periplocin in stem and leaf is probably transported by conducting tissue.

Numerical taxonomic studies were carried out in order to elucidate the taxonomic relationship among 17 species belonging to Paris. Eighteen characters including 10 morphological, 4 pollen morphological, 2 cytotalonomical and 2 habitat characters were used for the analysis. On basis of UPGMA clustering analysis, two subgenus and seven groups were recognized. The classification of the two subgenus was different from the opinion of subgenus Daiswa and subgenus Paris by Li Heng. The classification of sect. Dunnianae, sect. Axiparis and sect. Paris was correspondence with the classification of Li Heng. But sect. Fargesianae, sect. Marmoratae and sect. Thibeticae which were established based on the especially characters by Li Heng were put into cluster 2, cluster 3 and cluster 4.

Space breeding in medicinal plants is special characteristics in China. Compared with other plants, in spite of a relatively small number, Medicinal plants have more obvious characteristics and advantages. Research on medicinal plants has also been carried into all aspects, such as biological traits, physiology and biochemistry, genomics, as well as differences in chemical composition, and chemical composition analysis is also involved. However, compared with other plants, especially crops and vegetables, biological research is an obvious deficiency, that is mainly reflected in the insufficient genetics and breeding researches, the stability of genetic traits from generation to generation were not followed up and in-depth study in breeding areas was not carried out. If medicinal plants resources from space with the genetic stability good quality were selected, it would address the problem of lack of resources and ease the pressure on wild resources of medicinal plants. It would at the same time play an important role in promoting the development of medicinal botany space breeding and the implementation of modernization of traditional Chinese medicine.

To investigate the space environment on the role of licorice mutagenesis analysis of proteins.Licorice (Glycyrrhiza uralensis) seeds were carried by a recoverable satellite for 18 days (the average radiation dose in the flight recovery module was 0.102 m x d(-1), the orbit semidiameter 350 km, gravity 10(-6)). After return, The satellite-flown seeds and the unflown seeds (ground control) were planted in the fields of experimental farm. The leaves of each group were used for studying the effects of space flight on CAT, SOD activity, the protein content and electrophoresis.After the space flight, CAT, SOD activity of licorice increased in varying degrees, the difference was significant (P0.05), two types of enzyme activity of sample from Ordos were higher than that from Hangjinqi. The protein content of licorice increased in a certain extent, the difference was significant (P0.05), while protein electrophoresis also showed differences, weak new bands appeared.These results indicated that spaceflight has effect on protein of licorice, these changes may be used as a tool for accelerating the progress in G. uralensis breeding.

On-line ultra-performance liquid chromatography (UPLC) coupled with diode-array detection (UPLC/DAD) and electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) were used for separation, identification and structural analyses of saponins in Rhizoma Paridis saponins (RPS) and rat plasma after oral administration of RPS. Thirty steroidal saponins in RPS were identified by comparing their retention time, accurate mass measurement and positive and negative mass spectrometry data with that of reference compounds. The UPLC/Q-TOF-MS method was proved to be rapid and efficient in that 30 steroidal saponins, including three kinds of saponins (prototype, pennogenyl and diosgenyl saponins) were tentatively characterized within 6 min. After oral administration of RPS, 21 original saponins were absorbed in RPS-treated rat plasma. Our results indicated that UPLC/Q-TOF-MS is a rapid and effective tool for identification of a series of saponins at trace level. Copyright © 2010 John Wiley & Sons, Ltd.

To substantiate the effects of spaceflight on the glycyrrhizic acid-related gene mutation in Glycyrrhiza uralensis.Licorice (G. uralensis) seeds were carried by a recoverable satellite for 18 days (the average radiation dose in the flight recovery module was 0. 102 m x d(-1), the orbit semidiameter 350 km, gravity 10(-6)). After returned to the earth, the satellite-flown seeds and the un-flown seeds (ground control) were planted in the fields of experimental farm. The leaves of each group were used for studying the effects of space flight on the glycyrrhizic acid-related gene mutation including ITS sequence and beta-amyrine synthase gene.The ITS sequence of glycyrrhizic acid related gene showed no changes after spaceflight. While beta-amyrine synthase gene had some different points after spaceflight and the different points could get the expression.The results indicated that spaceflight induce genetic variation in G. uralensis and spaceflight could also have effects on glycyrrhizic acid-related gene mutation in G. uralensis. It may need to further research how the spaceflight induced the mutation of the glycyrrhizic acid related gene. The results suggested that recoverable satellite-flown condition could bring inheritable mutagenic effects on G. uralensis seeds and maybe used as a tool for accelerating the progress in G. uralensis breeding.

Fructus Aurantii, as a component of several compound formulae, shows many bioactivities. As is well known, the constituents of traditional Chinese medicines are very complex and multiple constituents are responsible for the therapeutic effect. However, the concrete compounds are difficult to confirm. Therefore, studies on the constituents absorbed into serum and their metabolites are necessary.To search for the active constituents in Fructus Aurantii.An ultra-performance liquid chromatography-photodiode array detector-quadrupole time-of-flight mass spectrometry (UPLC-PDA-Q-TOF/MS) method was established to analyse the aqueous extract in Fructus Aurantii and the constituents absorbed into blood. Compound identification was made by matching the empirical molecular formula with those of the reported compounds and UV spectra, and further elucidated by lower energy CID mass spectra.Twelve flavonoid O-glycosides were detected, and nine compounds were tentatively identified as polymethoxylated flavones. Six parent compounds were identified and four metabolites were observed in rat plasma, two of which were identified as naringenin glucuronide and hesperetin glucuronide, respectively.The approach developed has proved useful in the study of the active constituents in traditional Chinese medicines.

OBJECTIVE To investigate the space environment on the role of licorice mutagenesis analysis of proteins. METHOD Licorice (Glycyrrhiza uralensis) seeds were carried by a recoverable satellite for 18 days (the average radiation dose in the flight recovery module was 0.102 m x d(-1), the orbit semidiameter 350 km, gravity 10(-6)). After return, The satellite-flown seeds and the unflown seeds (ground control) were planted in the fields of experimental farm. The leaves of each group were used for studying the effects of space flight on CAT, SOD activity, the protein content and electrophoresis. RESULT After the space flight, CAT, SOD activity of licorice increased in varying degrees, the difference was significant (P

OBJECTIVE To optimize the culture condition of adventitious roots of Panax ginseng. METHOD The adventitious roots were obtained through tissue culture by manipulation of inoculum, various sucrose concentrations and salt strength. The contents of ginsenosides Re, Rb1 and Rg1 were determined by HPLC while the contents of polysaccharides were determined by ultraviolet spectrophotometry. RESULT The multiplication of adventitious roots reached the peak when the inoculum was 20 g x L(-1). The effects of sucrose concentration and salt strength on adventitious roots were observed. The contents of polysaccharides were higher when the medium contained more sucrose. 40 g x L(-1) sucrose was favorable for roots growth and biosynthesis of Re, while 30 g x L(-1) was favorable for the biosynthesis of Rb1 and Rg1. 3/4MS medium was benefit for the growth of adventitious roots and the biosynthesis of ginsenosides. The contents of polysaccharides were decreased with the increase of salt strength. CONCLUSION The results showed that inoculum, various sucrose concentrations and salt strength have significant influences on adventitious roots growth, secondary metabolite and polysaccharide synthesis in P. ginseng.

To study the affects of the different bioreactors on the Salvia miltiorrhiza adventitious root culture.Adventitious roots of S. miltiorrhiza were induced and in vitro cultured in bioreactors. The type of bioreactors was optimized and the kinetics of root growth, accumulation of the active ingredients was also investigated by HPLC.It showed that the 3-1 conical bubble bioreactor (CNBB) with 60 degrees taper favors achieved the highest active compounds accumulation in adventitious roots. The growth curve and secondary metabolites accumulation curve looks like "S" in CNBB bioreactor. The maximum adventitious roots biomass of 16.24 g x L(-1) (fresh weight) was obtained at 35 day. The highest content of tanshinone IIA (TA) and protocatechuic aldehyde (PA) reached 0.23 mg x g(-1) DW and 0.51 mg x g(-1) DW at 40 day, respectively.The 3-1 conical bubble bioreactor (CNBB) with 60 degrees taper favors was the best bioreactors for the accumulation of active compounds.

To study the chemical constituents in herbs of Paris verticillata.The compounds were isolated by column chromatography with silica gel and purified by Sephadex LH-20 and RP-HPLC. The structures were identified by means of NMR analysis.Nine compounds were isolated from the EtOAc extract and the n-BuOH extract of P. verticillata. Their structures were identified as beta-sitosterol (1), stigmasterol (2), daucosterol (3), beta-ecdysterone (4), 4-hydroxymethyl-gamma-butyrolactone (5), diosgenin-3-O-alpha-L-arabinofuranosyl (1--4)-[alpha-L-rhamnopyranosyl (1--2)]-beta-D-glycopyranoside (6), pennogenin-3-O-alpha-L-arabinofuranosyl(1--4)-beta-D-glycopyranoside (7), pennogenin-3-O-alpha-L-arabinofuranosyl (1--4)-[alpha-L-rhamnopy-ranosyl (1--2)]-beta-D-glycopyranoside (8), and pennogenin-3-O-alpha-L-rhamno-pyranosyl (1--4)-alpha-L-rhamnopyranosyl (1--4)-[alpha-L-rhamnopyranosyl (1--2)]-beta-D-glycopyranoside (9).Compounds 1-9 are isolated from P. verticillata for the first time, and compounds 3, 5 are isolated from the genus Paris for the first time. The compounds 6-9 showed certain inhibition activeness of LA-795 cells, especially, the effects of compounds 6, 8 and 9 were more significant.

To explore the distribution of cells infected by AD5-EGFP infused in different ways in the cochlea of guinea pig.AD5-EGFP was infused into the endolymphatic system through a hole on the lateral wall of the scala media or into the extralymphatic system through the round window membrane respectively. The infected cochlear cells confirmed by expression of EGFP were examined on the whole mount or cryostat sections.In the cochleae in which AD5-EGFP was infused into the extralymphatic system through the round window membrane, expression of EFGP could be found in the type I, IV and V fibrocyte of the stria vascularis, superlimbal cells of the spiral lip, cells in Ressenal membrane, spiral ganglion neurons in Rosenthal hole and cells lining the inner wall of scala vestibular and scala tympani, indicating that these cells were infected by adenovirus. None of the inner, outer hair or supporting cells was found to be infected in these cochleae. In the cochleae in which AD5-EGFP was infused into the endolymphatic system through a hole on the lateral wall of scala media, expression of EFGP could be found in supporting cells in the organ of Corti and lining cells of the scala media.Adenovirus5 is a good and effective vector for delivering genes into cells in guinea pig's cochlea. The scope of infected cells will be very different when the vector is applied to the cochlea through different infusion ways. No cells in the endolymphatic system would be infected if the vector is infused into the extralymphatic system.

Aim of study Croton oil is the fruit oil of Croton tiglium L., which is well known in folk medicine for the treatment of gastrointestinal (GI) diseases, including constipation, abdominal pain, peptic ulcer, and intestinal inflammation for a long period. This study was to investigate the pharmacological effect of croton oil on GI tract. Materials and methods The effect of croton oil on the smooth muscle contractions was investigated in vitro using the isolated rabbit jejunum model. Results Croton oil has a biphasic action contracting and relaxing intestinal tissue. At the concentrations of 20–80 μg/mL, croton oil produced a concentration-dependent increase in the amplitude and tension of muscle contractions, whereas at high concentrations (>200 μg/mL) it decreased the contractile amplitude and had no impact on the tension. Moreover, croton oil was less effective in increasing muscle amplitude and tension than Ach, confirming that the effect of croton oil on muscle contractions is not a simply stimulatory or inhibitory action, but a unique modulatory process depending on the concentration of croton oil. In addition, croton oil concentration-dependently suppressed the frequency of muscle contractions. On the other hand, atropine (10 μM) and 4-DAMP (10 μM) produced a significant inhibition of contractions caused by croton oil, while either hexamethonium (10 μM) or methoctramine (10 μM) was inactive, implying that the regulatory effects of croton oil on GI motility are mediated via the activation of M3 muscarinic receptor. Furthermore, muscle contractions induced by croton oil were dramatically reduced by verapamil (0.1 μM) but not by NE (1 μM), suggesting that the action of croton oil on GI motility is also mediated by Ca 2+ influx through L-type Ca 2+ channel. Conclusions The results suggest that croton oil possesses spasmogenic and spasmolytic properties and the regulatory effects of croton oil on GI motility are mediated via the activation of M3 muscarinic receptor and Ca 2+ influx through L-type Ca 2+ channel.

To study the chemical constituents from Centipeda minima.The constituents were isolated by column chromatography on silica gel, Sephadex LH-20 and HPLC-ODS, and identified by spectroscopic methods.Ten compounds, (-)-cis-chrysanthenol-O-beta-D-glucopyranoside (1), methy 3,5-dicaffeoylquinate (2), 3,5-di-O-caffeoyl quinicacid (3), tricin (4), 2-amino-4-methyl-pentanoicacid (5), 2-amino-3-phenyl-propionic acid (6), 4-amino-4-carboxychroman-2-one (7), brevilin-A (8), arnicolide C (9), arnicolide D (10) were isolated and identified.Compounds 1-7 were isolated from the plant for the first time.