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This study aimed to investigate the effect of Desmodium renifolium(Linn.) Schindl on ovariectomized rat model of osteoporosis and explore the underlying mechanism. Rats were randomized into a sham group, a model group, a Xianlin Gubao group, and low-, medium-, and high-dose D. renifolium groups. Bilateral oophorectomy was performed in other groups except sham group(removal of bilateral peri-ovarian adipose tissue). The sham group and model group were administrated with equal volume of 0.5% CMC-Na, Xianlin Gubao group with Xianlin Gubao, and D. renifolium groups with different concentrations of alcoholic extract of D. renifolium once a day for 14 weeks. The body weight of rats were recorded during the experiment. The levels of estradiol(E_2), 1,25-dihydroxyvitamin D_3 [1,25(OH)_2D_3], calcium(Ca), and phosphorus(P) in serum were determined after the administration. The femur microstructure was analyzed via micro-CT. RT-PCR and Western blot were employed to respectively determine the mRNA and protein levels of bone morphogenetic protein 2(BMP-2), Runt-related transcription factor 2(Runx2), and osterix(OSX) in the tibia of rats. The results indicated that D. renifolium significantly inhibited the weight growth, improved the uterus appearance, elevated the levels of E_2, Ca, P, and 1,25(OH)_2D_3 in serum, increased the number and reduced the fracture of bone trabeculae, ameliorated the bone microstructure parameters, and up-regulated the mRNA and protein levels of BMP-2, Runx2, and OSX. All the results demonstrated that D. renifolium had significant protective effect on the ovariectomized rat model of osteoporosis by regulating the BMP-2/Smads signaling pathway.

To study the non-flavonoids chemical constituents in water extract of Spatholobi Caulis. Some purification and analysis techniques like silica gel, D101-macroporous adsorptive resins, and Sephadex LH-20 column chromatographies as well as reversed phase high-performance liquid chromatography were used to isolate and analyze the phenolic acid esters and other type compounds from Spatholobi Caulis integrally. The structures of these compounds were identified by spectroscopic techniques such as nuclear magnetic resonance and high resolution mass spectrometries. Twenty-seven compounds, including phenolic acid, coumarin, lignan, terpene, alkaloid, and steroid compounds, were isolated from ethyl acetate and n-butanol fractions in water extract of Spatholobi Caulis, and they were identified as β-sitosterol(1), feruli acid methyl ester(2), syringaresinol(3),(+)-medioresinol(4),(+)-epipinoresinol(5), p-acetylphenol(6), bolusanthin Ⅳ(7), evofolin B(8), salicylic acid(9), trans-p-hydroxy-cinnamic acid(10), abscisic acid(11), m-hydroxyphenol(12), C-veratroylglycol(13), p-hydroquinone(14), 8,9-dihydroxymegastigma-4,6-dien-3-one(15), p-hydroxybenzoic acid(16), 6,9-dihydroxymegastigma-4,7-dien-3-one(17), protocatechuic acid(18), protocatechuic acid methyl ester(19), 5,7-dihydroxycoumarin(20), isolariciresinol(21), nicotinic acid(22), daucosterol(23),(+)-pinoresinol(24), stigmasterol(25), allantoin(26) and koaburaside(27), respectively. Furthermore, compounds 2-15, 19-22, 24 and 26 were isolated from genus Spatholobus for the first time.

The chemical compounds in water extract of Spatholobi Caulis were further studied. The compounds were systematically isolated and purified by using various separation and analysis techniques including silica gel, macroporous adsorptive resins and Sephadex LH-20 column chromatographies, as well as reversed phase high-performance liquid chromatography(RP-HPLC). Twenty-three flavonoids and one chromone were identified by the spectroscopic analysis techniques combining their physicochemical properties, they were identified as isoduartin(1), sativan(2), 8-O-methylretusin(3), 7-hydroxydihydroflavone(4), odoratin(5), butesuperin A(6), biochanin A(7), 3'-methoxydaidzein(8), 7-hydroxychromone(9), calycosin(10), naringenin(11), dihydrocajanin(12),(6 aR,11 aR)-maackiain(13), 2'-hydroxygenistein(14),(6 aR,11 aR)-medicarpin-3-O-glucopyranoside(15),(-)-epiafzelechin(16),(-)-catechin(17),(-)-epicatechin(18), 4',8-dimethoxy-7-O-β-D-glucopyranosylisoflavone(19), ononin(20),(-)-gallocatechin(21), rutin(22), daidzin(23) and sphaerobioside(24). Compounds 4, 6, 8, 9, 11, 12, 14-16, 19 and 22-24 were isolated from Spatholobi Caulis for the first time.

A method was established for simultaneous determination of 21 active constituents including flavanols, isoflavones, flavonols, dihydroflavones, dihydroflavonols, chalcones, pterocarpan, anthocyanidins and phenolic acids in Spatholobi Caulis by ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry(UFLC-QTRAP-MS/MS). Then, it was employed to analyze and evaluate the dynamic accumulation of multiple bioactive constituents in Spatholobi Caulis. The chromatographic separation was performed on a XBridge®C_(18)(4.6 mm×100 mm, 3.5 μm) at 30 ℃ with a gradient elution of 0.3% formic acid aqueous solution-methanol, and the flow rate was 0.8 mL·min~(-1), using multiple-reaction monitoring(MRM) mode. A comprehensive evaluation of the multiple bioactive constituents was carried out by gray correlation analysis(GRA). The 21 target components showed good linearity(r>0.999 0) in the range of the tested concentrations. The average recovery rates of the 21 components were from 97.46% to 103.6% with relative standard deviations less than 5.0%. There were differences in the contents of 21 components in Spatholobi Caulis at diffe-rent harvest periods. Spatholobi Caulis had high quality from early November to early December, which is consistent with the local tradi-tional harvest period. This study reveals the rule of the dynamic accumulation of 21 components in Spatholobi Caulis and provides basic information for the suitable harvest time. At the same time, it provides a new method reference for the comprehensive evaluation of the internal quality of Spatholobi Caulis.

HPLC analysis was performed on a Phenomenex PS C₁₈（4.6 mm×250 mm, 5 μm）column using methanol -0.2% formic acid (30:70) at a flow rate of 0.8 mL·min⁻¹. The column temperature was 30 °C and the detection wavelength was set at 335 nm. The injection volume was 10 μL. The HPLC fingerprint of Desmodium styracifolium was established with 10 common peaks, and 5 of them were identified as vicenin-1, schaftoside, isoorientin, isoschaftoside and isovitexin, respecivetly. The fingerprints of 21 batches of D. styracifolium samples were analyzed with similarity evaluation, cluster analysis, principal component analysis and partial least squares discriminant analysis. There was no significant difference among the quantitative results of these five ingredients verified by external standard method (ESM) and quantitative analysis of multi-components by single marker (QAMS) method. The application of fingerprint, pattern recognition combined with QAMS can provide more comprehensive references for the quality control and evaluation of D. styracifolium., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

In this study, complex enzymes combined with ultrasonic extraction technology（MC） were used, to select optimal extraction combinations by single factor and orthogonal test, with Hedysarum polysaccharides yield and content as the comprehensive indexes. The components, physicochemical properties and antioxidant activity of Hedysarum polysaccharides from complex enzyme combined with ultrasonic extraction（HPS-MC）and the Hedysarum polysaccharides from hot water extraction（HPS-R）were analyzed. The results showed that：complex enzymes had significant effect on the yield and content of Hedysarum polysaccharides, and the ultrasonic power could significantly improve the content of Hedysarum polysaccharides. The optimum technological parameters were as follows: complex enzyme ratio 1:1, ultrasonic power 105 W, ultrasonic time 60 min, and enzymatic hydrolysis pH 5, achieving （14.01±0.64）% and （92.45±1.47）% respectively for the yield and content of Polysaccharides. As compared with HPS-R, the molecular weight, absolute viscosity and protein content of HPS-MC were decreased, while the content of uronic acid was increased. In the antioxidant system, the concentration of polysaccharide was within the range of 1-7 g·L⁻¹; the antioxidant activity of HPS-MC was higher than that of HPS-R, and HPS-MC (80%) with the lowest molecular weight showed a significant dose effect relationship with the increase of the experimental concentration. In conclusion, MC is a simple, convenient, economical and environmentally friendly extraction technology, and the Hedysarum polysaccharides extracted by this method have obvious antioxidant activity., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Spatholobi Caulis, the vine stem of Spatholobus suberectus and widely used in China and Southeast Asian nations, has the effects on nourishing the blood and promoting blood flow, regulating menstruation and relieving pain, and invigorating the nerves. Through consulting the herbal textual and local chronicles, we summarized the original textual research and medicinal evolution on Spatholobi Caulis to analyze the changes of varieties in different historical periods. Further, the major active ingredient in Spatholobi Caulis was discussed. According to the literature to date, 60 flavonoids compounds have been isolated and could be divided into isoflavones, dihydroflavones, flavanols, dihydroflavonols, procyaninides, chalcones, pterocarpans, isoflavanols, isoflavanones and aurone according to their molecular structures. These indicative ingredients in Spatholobi Caulis showed pharmacological activities on regulation of the blood system, anti-tumor, anti-oxidation, anti-virus, anti-bacteria and inhibition of melanin deposition. This review will provide reference and basis for the sustainable use of resources and industry development on Spatholobi Caulis., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Objective: To determine the effect of Radix Angelicae Sinensis and Radix Hedysari (RAS-RH) on radiosensitivity of human liver cancer H22 cells to heavy ion ¹²C⁶⁺and its possible mechanism., Methods: The experiment involved a comparison of proliferation of H22 cells (detected by CCK-8 assay) between four groups: control,drug (RAS-RH),radiation,and combination (RAS-RH+radiation). H22 cells were treated with different doses of radiation alone or radiation followed by RAS-RH. The radiation enhancement effect of RAS-RH on H22 was detected by Colony forming assay. The effect of RAS-RH on the apoptosis of H22 cells was detected by flow cytometry. The influence of RAS-RH on the expression levels of related protein Survivin and Caspase-9 was investigated by Western blot., Results: RAS-RH inhibited the proliferation of H22 cells,with a time and dose dependency [inhibitory concentration 20% (IC 20 )=(117.60±2.15) mg/L]. The survival rate of H22 cells decreased significantly with the increase of heavy ion ¹²C⁶⁺. The two survival curves produced by the Graph Pad Prism 5.0 software were clearly separated. The combination group demonstrated smaller shoulder area at low dosage and lower survival rate of cells compared with radiation group,with a sensitization enhancement ratio ( SER ) of 1.39±0.07. The combination group (100 mg/L RAS-RH+2 Gy) had higher apoptosis rate and Caspase-9 protein expression level,and lower Survivin protein expression level,compared with other 3 groups ( P <0.01)., Conclusion: RAS-RH has radiation sensitization effect on human hepatocellular carcinoma H22 cells. The mechanism may be related to down-regulation of Survivin protein expression and up regulation of Caspase-9 protein expression.

To establish a HPLC-MS/MS quantitative method for analysis of 8 constituents in rat bile. The method was applied in the biliary excretion study after oral administration of Desmodii Styracifolii Herba extract. The rat bile samples were collected during 0-1, 1-2, 2-4, 4-6, 6-8, 8-12, 12-24 h. Diamonsil C₁₈column (4.6 mm×150 mm, 5 μm) was adopted and eluted with methanol and 0.01% acetic acid in a gradient mode. The flow rate was 0.8 mL•min⁻¹, and the column temperature was set at 40 ℃. The detection was carried out by a triple quadrupole linearion trap mass spectrometer in the negative ion mode with an electrospray source. Multiple reactions monitoring (MRM) mode was employed. The calibration curves showed a good linearity, with correlation coefficients (r≥0.991 5) for all of the analytes within the concentration range. The intra-day and inter-day precisions (RSD) were both less than 15%, and the accuracies (RE) ranged from -15% to 15%. The method had a good stability, and was suitable for the content determination of 8 constituents in rat bile. The extraction recoveries of the 8 analytes were more than 63.2%, and the RSD of IS-normalized matrix factors were no more than 15%, which met the requirements for analysis. According to the biliary excretion study, the bile excretion rates of the 8 analytes were relatively low, with great difference among the individuals. The results showed that the established method was simple, selective and specific, thus could be applied in the biliary excretion study of the 8 analytes in rat bile., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

The research on distribution and quality suitability division of Desmodium styracifolium were formulated by Maxent and ArcGIS model based on the content of schaftoside and polysaccharide of D. styracifolium and its field research in the south and southwest areas of China (Guangdong, Guangxi, Hainan and Yunnan), and the most suitable habitats of distribution suitability and quality suitability were screened. The distribution suitability results indicated that average air temperature in April,mean temperature of coldest quarter, soil type, coldness index were found as the four dominant factors contributing to the plant distribution. The quality suitability results indicated that: ①Polysaccharide content and precipitation in April show significant positive correlation;Schaftoside content and mean temperature of April, mean temperature of coldest quarter show significant negative correlation. Schaftoside content shows significant negative correlation with the precipitation in October and November and the sunshine duration in April and May, while there is a significant positive correlation between schaftoside content and precipitation in April and temperature seasonality standard deviation, and a highly significant positive correlation was found between schaftoside content and precipitation in February and March. ②The quality zoning map was drawn depend on general content of polysaccharide and schaftoside as the index of quality. And this research provides scientific location basis for the production regionalization, cultivation bases selection and directive breeding of D. styracifolium., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

In order to explore the adaptive capacity of Alhagi sparsifolia to soil salinity (mild, mo-derate, severe saline soil), the distribution, adsorption and translocation characteristics of Na + , K + , Ca 2+ , Mg 2+ in different organs of A. sparsifolia, which is a dominant native vegetation in southern edge of Tarim basin, were investigated. The results showed that when grown on a mild and moderate saline soil, Na + distribution in different organs of A. sparsifolia followed the order of stem≈thorn>leaf>root; when grown on a severe saline soil, Na + distribution followed leaf>stem≈thorn>root. The distribution of Ca 2+ and Mg 2+ followed leaf >thorn>stem>root. With the increase of soil salt level, Na + content in different organs of A. sparsifolia increased, while K + content in leave decreased; meanwhile the increasing soil salt level significantly decreased the K + /Na + ratio in both root and leave, as well as the Ca 2+ /Na + and Mg 2+ /Na + ratios in all organs. The selective transportation coefficients for Ca 2+ -Na + and Mg 2+ -Na + followed order of stem-leaf>stem-thorn>root-stem under saline habitats. Based on the integrated analysis, in order to adapt to salinity habitat, A. sparsifolia could use multiple organs to accumulate Na + , depending on soil salt level. Na + could accumulate in stem and thorn at a relative low level, but in leaves at high levels. Besides, Ca 2+ and Mg 2+ could be the inorganic osmotic adjustment substances for A. sparsifolia to adapt the saline environment.

This paper was aimed to compare the effect of Buzhong Yiqi decoction containing Hedysari Radix or Astragali Radix on anti-immunosenescence effects in spleen lymphocytes of senescence accelerated mouse 8 (SAMP8). The effect of the serums on the proliferation of spleen T lymphocytes in SAMP8 mice induced by ConA was tested by MTT. The effect of the serums on the T lymphocytes subsets of SAMP8 mice was measured by flow cytometry. ELISA was used to detect the level of IL-2 and IFN-γ in the culture supernatants of spleen lymphocytes. The effect of the serums on the expression of CD28 mRNA in spleen T lymphocytes was detected by fluorescent quantitative PCR. Western blot was used to detect the expression of CD28 protein in spleen T lymphocytes of SAMP8 mice. Both the serums of Buzhong Yiqi decoctions containing Hedysari Radix or Astragali Radix improved the proliferation of T lymphocytes in SAMP8 mice. Both the serums had no obvious effect on the differentiation of spleen T lymphocytes'subsets in SAMP8 mice. Both the serums increased the content of IL-2 and INF-γ in the culture supernatants of spleen lymphocytes. And for the content of IL-2, the serum of Buzhong Yiqi decoction with Hedysari Radix was better（P<0.05）. Both the serums improved the expression of CD28 mRNA in spleen T lymphocytes of SAMP8 mice. And the effect of Hedysari Radix group was better than that of Astragalus Radix group（P<0.05）. Both the serums improved the expression of CD28 protein in spleen T lymphocytes of SAMP8 mice. The role of the serums containing Buzhong Yiqi decoction with Astragalus Radix and the decoction with Hedysari Radix in anti-immunosenescence was through the effect of the CD28. And the effect of Hedysari Radix group was better than that of Astragalus Radix group on improved the expression of CD28 mRNA in T lymphocytes of SAMP8 mice. Astragalus Radix and Hedysari Radix could swap in the aspect of anti-immunosenescence., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

To study the anti-radiation effect and mechanism of ethanol extracts from Spatholobus suberectus and its active component catechin, ICR mice were exposed to 6Gy irradiation and randomly divided into normal group, model group, positive control group (amifostine, 43.6 mg•kg⁻¹, iv 30 min before irradiation), SSD group (10, 20, 40 g•kg⁻¹) and catechin group (50, 100, 200 mg•kg⁻¹). The mice were administered the appropriate drugs once a day after irradiation for 28 consecutive days. Blood samples were collected from the tail end and the number of peripheral blood cells was counted before irradiation and on day 1, 3, 7, 14, 21 and 28 using a microcell counter. Changes of thymus and spleen index of mice on day 7 were observed. The serum SOD, GSH-Px activity and MDA level were detected by the colorimetric method. The colony forming ability of bone marrow hematopoietic progenitor cells on day 7 was detected by semi solid culture method. The HE staining was adopted to observe the pathological changes. The apoptosis of bone marrow cells was detected by flow cytometry. The expression of cleaved caspase-3 and Bax of bone marrow cells were measured separately by western-blotting and immunohistochemistry method. SSD and catechin can both significantly revert the irradiated-induced decline in hematological parameters (RBC, WBC, PLT, Hb), improve thymus and spleen index, significantly enhance serum SOD and GSH-Px activity and decrease the MDA level. The proliferation and differentiation of hematopoietic progenitor cells in bone marrow were promoted, the apoptosis of bone marrow cells was significantly up-regulated and the expression of cleaved caspase-3 and Bax was significantly reduced in SSD and catechin group. SSD and catechin have significant anti-radiation effect and its mechanism may be related to hematopoietic promoting, antioxidant and anti-apoptotic effects., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

An HPLC method was established for the determination of adenosine, γ-aminobutyric acid (GABA) and six flavonoids (calycosin-7-glucoside, ononin, calycosin, isoliquiritigenin, formononetin and medicarpin） in Radix Hedysari. The samples were extracted with methanol by refluxing for 4 h. The HPLC-DAD was performed on a Diamonsil C(18) column (250 mm × 4.6 mm, 5 μm) with acetonitrile-water as the mobile phase. The column temperature was at 40 ℃ and the flow rate was 1.0 m L·min(-1), while the temperature of drift tube was 110.5 ℃ and the nebulizing gas flow was 3.1 L·min-1 for the ELSD system. The results showed all the eight components had good linear relationships (r(2) =0.992 8-1.000 0) in the range of the test concentration. The RSD of precision, stability and repeatability were less than 2%.The average recovery rates were 96.78%-103.45%, and RSD were 0.29%-1.61%.The index component contents of Radix Hedysari form 24 different origins were determined and used as variable factors in clustering analysis. The results were classified into 2 groups basically in accordance with the regional cluster. And the consequence was in consistent with the results of principal component analysis. This HPLC method is simple, shows good sensitive and accurate, and provides the experimental basis for multi-index control of Radix Hedysari. Clustering analysis for Radix Hedysari quality control has a certain reliability and objectivity.

Astragalus membranaceus var. mongholicus and Hedysarum polybotrys belong to different genera, but have similar drug efficacy in traditional Chinese medicine theory, and H. polybotrys was used as the legal A. membranaceus var. mongholicus previously. In this study, similarities and differences between them were analyzed via their ITS/ITS2 fragments information. The ITS (internal transcribed spacer) regions were amplified using polymerase chain reaction and then sequenced in two-way. The alignment lengths of ITS regions were 616 bp, in which 508 loci were consistent, and 103 loci were different, accounting for 82.47% and 16.72% of the total ITS nucleotides in length, respectively. As genotype determines phenotype, 1HNMR-based metabolomic approach was further used to reveal the chemical similarities and differences between them. Thirty-four metabolites were identified in the 1H NMR spectra, and twenty-seven metabolites were the common components. Amino acids, carbohydrates and other primary metabolites were similar, while a large difference existed in the flavonoids and astragalosides. This study suggests that A. membranaceus var. mongholicus and H. polybotrys show similarities and differences from molecular and chemical perspectives, which has laid a foundation for elucidating the effective material basis of drug with similar efficacy and resources utilization.

The dynamic changes of germination percentage, germination potential, thousand-seed weight, antioxidase activity in Desmodium styracifolium seeds with different storage time were tested, and electrical conductivity, contents of soluble sugar, soluble protein, starch in seed leach liquor were also determined in order to reveal the mechanism of seed deterioration. The results as the following. (1) The germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds declined, while the seed coat color darkened with the extension of storage time. (2) The activities of superoxide dismutase (SOD) and peroxidase (POD) decreased with the prolongation of storage period. The SOD activity declined fastest in 1,095-1,185 d of storage, while the POD activity declined significantly in 365-395 d of storage. (3) The electrical conductivity and the contents of soluble sugar, starch in seed leach liquor increased, while the content of soluble protein declined with the extension of storage time. (4) Correlation analysis indicated that the germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds have a significantly positive correlation with SOD and POD activity, while have a significantly negative correlation with the electrical conductivity, contents of soluble sugar and starch. It can be concluded that during the storage of D. styracifolium seeds, physiological and biochemical changes including decrease in antioxidase activity, rise in electrical conductivity, degradation effluent of soluble sugar and starch, degradation of soluble protein were the main factors leading to the seed deterioration.

A modified QuEChERS (quick, easy, cheap, effective, rugged and safe) method followed by liquid chromatography-tandem mass spectrometric analysis was developed for the determination of dimethyl yellow and diethyl yellow in yuba and dried beancurd. Yuba and dried beancurd samples were soaked by deionized water, then acetonitrile was added to extract the analytes. Sodium chloride and anhydrous magnesium sulfate were added for liquid-liquid separation. The extracts were cleaned-up by dispersive solid-phase using N-propyl diethylamine. The analytes were separated by liquid chromatography and determined by mass spectrometry. External standard method was used for quantification. The recoveries of dimethyl yellow were in the range of 73.5%-84.5% at spiked levels of 0.3, 1 and 10 kg/kg and the recoveries of diethyl yellow were in range of 70.5%-81.2% at spiked levels of 0.1,1 and 10 µg/kg; relative standard deviations of the method were lower than 11%. The limit of detection and the limit of quantification of dimethyl yellow were 0.1 µg/kg and 0.3 µg/kg, respectively; the limit of detection and the limit of quantification of diethyl yellow were 0.05 µg/kg and 0.1 µg/kg, respectively. This method can be used in rapid screening and quantitative analysis of dimethyl yellow and diethyl yellow in yuba and dried beancurd.

Objective: To establish the quality standard of Entadae Semen, and provide scientific basis for its quality control., Method: TLC and HPLC were used for qualitative identification and quantitative analysis of phaseoloidin and entadamide A-O-β-D-glucopyranoside in Entadae Semen. The test of water content, ash and ethanol-soluble extractives of Entadae Semen was carried out according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition)., Result: The TLC was well separated with clear spots. The linear range of phaseoloidin was between 0.014 to 2.747 g x L(-1) (r = 0.999 6, n = 9) with an average recovery rate of 101.06% (RSD 0.90%, n = 6); the linear range of entadamide A-O-β-D-glucopyranoside was between 0.002 to 0.452 g x L(-1) (r = 0.999 7, n = 9) withan average recovery rate of 101.52% (RSD 1.09%, n = 6). The content of phaseoloidin in sample is between 5.12% to 9.24%, entadamide A-O-β-D-glucopyranoside is between 0.55% to 2.17%, alcohol-soluble extracts is between 30.9% to 45.2%, water is between 6.6% to 8.6%, and total ash is between 2.4% to 2.9%., Conclusion: The established standard is acceptable for quality control of Entadae Semen.

The spectrum-effect relationship on anti-hepatic fibrosis effect of Radix Hedysari was explored based on high performance liquid chromatographic technique. Hepatic fibrosis was induced in mice by administering a subcutaneous injection of carbon tetrachloride-peanut oil (4:6, v/v) continuously for 35 d at a interval of 5 d (0.1 mL/10 g). And at the same time of modeling, the different extracts of Radix Hedysari were administered orally once daily at a dose of 10 g/kg. The ethanol extract of Radix Hedysari was specified to be most effective on anti-hepatic fibrosis by determining the levels of alanine aminotransferase, aspartate transami- nase, total-protein, albumin, albumin/globulin (ALT, AST, TP, ALB, and A/G) in serum and relative liver weight. Subsequently, the grey relational degree analysis and partial least squares analysis were employed to reveal the correlation between chromatographic fingerprint of ethanol extract of Radix Hedysari from 10 different geographical origins and its anti-hepatic fibrosis efficacy. The results suggest that most chemical constituents of Radix Hedysari have a high correlation with the effect of anti-hepatic fibrosis (> 0.8), which indicates that the effect is related to the various components in Radix Hedysari. Adenosine, calycosin and ononin in ethanol extract of Radix Hedysari have been identified separately among which adenosine and calycosin made the great contribution to the anti-hepatic fibrosis effect.

Objective: To observe changes of [Ca2+]i concentration and CaM, CaMK II and p-CaMK II of Ca2+/CaMK II signaling pathways in skeletal muscle tissue of rats with spleen-qi deficiency and intervention of Sijunzi decoction and extract of Hedysarum polybotrys., Methods: Rats were randomized into four groups: normal control group, spleen-qi deficient model group, extract from Hedysarum polybotrys group and Sijunzi decoction group, ten rats in each group. After the spleen-qi deficient models were built by comprehensive application of rhubarb, exhaustive and hungry methods, and treatment groups were treated with extract from Hedysarum polybotrys at 6 g/(kg . d) or Sijunzi decoction at 20 g/(kg . d) for 21 d. Then, general existence,gastrointestinal hormones GAS and MOT levels, and activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase of skeletal muscle were evaluated. Also, confocal laser technology was used to test cellular[Ca2+]i concentrations in skeletal muscle and Western blotting technique was used to test CaM, CaMK II and p-CaMK 11 expression in intestinal tissue of spleen-qi deficient model rats., Results: Compared with normal group, general condition was poor, levels of GAS and MOT decreased (P <0. 01), activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase, [Ca2+]i concentration as well as expression of CaM, CaMK II and p-CaMK II in skeletal muscle decreased significantly (P < 0. 01) in spleen-qi deficienct model rats. Compared with model group, general condition improved significantly, as well as level of MOT in intestinal increased (P <0. 05) in the rats of extract from Hedysarum polybotrys group and Sijunzi decoction group,while level of GAS increased in intestinal(P <0. 05) in the rats of Sijunzi decoction group; Moreover, activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase as well as [Ca2+]i concentration and expression of CaM and CaMK II in skeletal muscle tissue increased (P < 0. 05) in the rats of extract from Hedysarum polybotrys group and Sijunzi decoction group, while p-CaMK II in skeletal muscle tissue increased in the rats of Sijunzi decoction group (P < 0. 05)., Conclusion: Sijunzi decoction and extract of Hedysarum polybotrys can be applied to treat spleen-qi deficiency syndrome through the mechanism of regulating GAS and MOT secretion and raising expression of Ca2+ /CaM signaling pathways key factors in skeletal muscle tissue. Sijunzi decoction has the better effect

In this study the chemical constituents of the higher polar sustances from Desmodium caudatum were investigated.The compounds were isolated by using column chromatographies over silicagel, polyamide, ODS, Sephadex LH-20, and preparative HPLC. The structures of these compounds were identified on the basis of NMR and MS spectra. Thirteen compounds were obtained and their structures were identified as vanillin(1), loliolide(2), indole-3-carboxaldehyde(3), salicylic acid(4), swertisin(5), saccharumoside C(6), isosinensin (7), kaempferol 3-O-β-D-glucopyranoside-7-O-α-L-rhamnopyranoside (8), isovitexin (9), vitexin (10), nothofagin(11), resveratroloside (12), and 2"-α-rhamnopyranosyl-7-O-methylvitexin (13). Except for compound 5, the remaining compounds were isolated from D. caudatum for the first time. Compounds 2, 3, 6-8, 11-13 were separated from the genus Desmodium for the first time.

Objective: Through researching the relationship among osteoporosis and inflammatory reaction besides angiogenesis, to compare pharmacological differences between icariin and genistein to inhibit bone loss., Methods: 6 months old female SD rats were randomly divided into SHAM group, model group, ICA group, GEN group and E group. The bone mineral density of total, femur and lumbar, serum OC, TRACP 5b, IL-6 and VEGF, biomechanics of femur and tibia microarchitecture were analyzed., Results: Compared with SHAM group, model group of body weight, uterine weight, bone mineral density of total, femur and lumbar, serum OC, TRACP 5b, IL-6 and VEGF, biomechanics of femur and lumbar and tibia microarchitecture were significantly changed (P < 0.05). Compared with model group, ICA group of body weight, bone mineral density of total and femur, serum TRACP 5b and femural biomechanics were significantly changed (P < 0.05). GEN group of bone mineral density of total, femur and lumbar, serum OC, TRACP 5b, IL-6 and VEGF, biomechanics of femur and lumbar and tibia microarchitecture were significantly changed (P < 0.05)., Conclusion: Icariin inhibits bone loss on model rat through suppressing bone resorption. Genistein prevents bone loss on model rat by the pathway of inhibiting inflammatory reaction, activating angiogenesis, enhancing bone formation and inhibiting bone resorption. Moreover, pharmacological activity of genistein is more potential than icariin.

Objective: To investigate the bioactive constituents against hepatic fibrosis from the roots of Phyllodium pulchellum., Methods: The chemical constituents of Phyllodium pulchellum roots were obtained by various chromatographic technologies and identified by several spectroscopic methods., Results: Ten compounds were elucidated as 3,5,2',4'-tetrahydroxy-2",2"-dimethylpyrano-[5",6",7,8] -flavanone (1), yukovanol (2), citflavanone (3), 8-prenylated 5,7,3',4'-tetrahydroxyflavanone (4), pulchelstyrene A (5), pulchelstyrene B (6), pulchelstyrene D (7), 3-indolcarbaldehyde (8), 3-methoxyindole (9) and p-hydroxybenzoic acid (10). The effects to inhibit the proliferation of activated HSC-T6 cells of all isolated compounds were also evaluated., Conclusion: All compounds are isolated from this plant for the first time except for compounds 5 - 7. Compounds 2,4,5 and 6 can inhibit the proliferation of activated HSC-T6 cells in vitro.

In order to control the quality of Vigna radiata, the quality control method and standard were established in this study. The tests of water content, ash and ethanol-soluble extractives of V. radiata were carried out according to the methods recoded in appendix of Chinese Pharmacopeia (2010 edition, volume 1). The TLC method was established by using vitexin and isovitexin as references, and a mixture of acetate-method-water (10: 1.7 : 1.3) as the developing solvent system on GF254 thin layer plate. The contents of vitexin and isovitexin were determined by HPLC on a Prevail C18 (4.6 mm x 250 mm, 5 microm) column, using acetonitrile: water (23 : 77) as mobile phase at a flow rate of 1.0 mL x min(-1). The column temperature is 30 degrees C and the detection wavelength is 337 nm. As a result, vitexin, isovitexin and the other constituents were well separated on TLC detected under the UV light (254 nm). The methodology validation for the assay of vitexin and isovitexin presented that they were in good linear correlation in the ranges of 6.12-98 mg x L(-1) and 6.85-109.6 mg x L(-1), with the regression equations of Y = 46.213X - 7.100 (r = 1.000) and Y = 54.515X + 6.829 (r = 1.000), and the average recoveries were 98.2% (RSD 1.9%) and 97.2% (RSD 0.79%), respectively. The content ranges of vitexin and isovitexin from 25 different batches of V. radiata were 1.076-2.062 mg x g(1) and 1.127-2.303 mg x g(-1), respectively. suggesting that the qualities of V. radiata are relatively stable. The ethanol-soluble extractives, water content and total ash of 25 samples varied in the ranges of 13.27% - 18.46%, 9.59% - 12.43% and 2.63% - 3.53%, respectively. All of the above data proved that the established quality of control method V. radiata is specific and accurate, which can be used for the quality control of this drug.

Objective: To study the chemical constituents of Desmodium caudatum., Methods: Silica column chromatography, Sephadex LH-20 column chromatography and recrystallization were used to separate and purify the chemical composition of Desmodium caudatum. Their chemical structures were identified by infrared spectrum (IR), mass spectrum (MS), nuclear magnetic resonance (NMR) and other physicochemical methods., Results: Twelve compounds were isolated and identified as lacceroic acid(1), gheddic acid(2), stigmasterol(3), betulin(4), citrusinol(5), yukovanol(6), kaempferol(7), protocatechuic acid(8), sophocarpine(9), matrine(10), N, Ndimethyltryptamine(11) and 5-hydroxy-N,N-dimethyltryptamine(12)., Conclusion: Compounds 1, 2, 4 and 8-12 are isolated from this plant for the first time.

The chemical constituents isolated from Desmodium species (Leguminosae) included terpenoids, flavonoids, steroids, alkaloids compounds. Modem pharmacological studies have showed that the Desmodium species have antioxidant, antibacterial, anti-inflammatory, hepatoprotective, diuretic, antipyretic, analgesic and choleretic activity. This article mainly has reviewed the research advances of chemical constituents and biological activities of Desmodium species since 2003.

SHG was sulfated by chlorosulfonic acid-pyridine method, and six samples which we got were prepared in different reaction conditions. There is a characteristic absorption peak near 260 nm in UV spectra and there are two characteristic absorption peaks near 1240 cm(-1) and 810 cm(-1) in the FT-IR. Degree of sulfation (DS) was calculated by elemental analysis and turbidimetry. Under the same conditions the absorption peaks become strong with the DS increase. The anticoagulant activity of SHG and sulfated modification samples was evaluated by the classic coagulant assays of prothrombin time (PT), activated partial thrombin time (APTT) live enzymes, and plasma thrombin time (TT). Results show that sulfated SHG has a good anticoagulant activity in vitro, and DS increased activity within a certain range.

Treatment of peripheral nerve injury is a major challenge in clinical practice. With advances in molecular biology and development of microsurgical techniques and tissue engineering, peripheral nerve repair procedures have been greatly improved. In the last 10 decades, most treatments for peripheral nerve injury in animal models have achieved histological and functional recovery, the treatments in humans, however, produce insufficient recovery, especially for proximal nerve injury. Increasing attention has been paid to the Traditional Chinese Medicine (TCM) for promoting peripheral nerve regeneration, since these remedies often display effective clinical outcome, minor side effects and effectiveness for multiple targets. Although TCM has complex ingredients and the specific pharmacological mechanisms for their effectiveness are still unclear, an effective clinical outcome is welcomed by many clinicians. In the past 20 years, we have made a series of detailed studies including the toxicity tests, pharmacodynamic tests, pharmacological experiments etc, about a new traditional formula which mainly contains the Radix hedysari, Epimedium etc. RESULTS have shown that this formula is safe to be used in both animals and humans with no toxicity and adverse effect, and systemic administration of this formula could enhance the peripheral nerve regeneration.

Objective: To optimize extraction techniques for genistein in Flemingia macrophylla by using response surface method., Methods: Based on the single factor experiment, extraction temperature, extraction time and solvent ratio served as the response factor while the extraction rate of genistein served as the response value. Based on multiple linear regression and binomial expression, response surface method was employed to optimize the extracting process, and predicative analysis was conducted., Results: The effects of factors in the descending order were as follows: extraction temperature, extraction time and solvent ratio. The optimal extraction technology was as follows: extraction temperature 80 degrees C, extraction time 3.5 h and solvent ratio 20 mL/g. Under these conditions, the extraction rate of genistein reached 0.161 mg/g., Conclusion: The results are agreed with model predictions. The technology can be used as the extraction process of genistein in Flemingia macrophylla.

Phytochemical investigation was carried out on the seeds of Vigna umbellata. The 70% ethanol extract of the seeds of V. umbellata was subjected to silica gel, Sephadex LH-20, ODS column chromatographies and preparative HPLC. The structures of the isolated compounds were elucidated on the basis of NMR and ESI-MS spectroscopic data Eight compounds were obtained and identified as carboxyatractyligenin (1), 2beta-O-beta-D-glucopyranosyl-15alpha-hydroxy-kaur-16-ene-18,19-dicarboxylic acid (2), 2beta-O-(beta-D-glucopyranosyl) atractyligenin (3), 3R-O-[beta-L-arabinopyranosyl-(1-6) -beta-D-glucopyranosyl] oct-1-ene-3-ol (4), (6S, 7E, 9R) -roseoside (5), liriodendrin (6), resveratrol (7) and maltol (8). Compounds 1-7 were isolated from Vigna genus for the first time, and compound 8 was isolated from V. umbellata for the first time.

Dalbergiae Odoriferae Lignum is a traditional Chinese medicine (TCM) commonly used for promoting blood circulation, relieving pain and removing blood stasis. Volatile oil and flavonoid compounds are two main chemical constituents of Dalbergiae Odoriferae Lignum. Modern pharmacological studies show that Dalbergiae Odoriferae Lignum has many effects, such as relaxing blood, increasing blood flow of coronary, anti-oxidation, anti-inflammation and antitumor. Dalbergiae Odoriferae Lignum, as a characteristic TCM with the potential of further development, is generally compatible with other TCMs to treat cardio-cerebral vascular diseases. This article summarizes studies on chemical composition, pharmacological action, pharmacokinetic procfile in vivo and TCM compatibility in recent years, in order to provide references for further studies.

Objective: To study the effect of flavonoids from Hedysari Radix on pulmonary functions of pulmonary fibrosis rat and its mechanism., Methods: 72 Wistar rats were randomly divided into 6 groups: blank control group, model group, prednisone group, Hedysari Radix flavonoids low, medium and high dosage group. The rat model was established by propelling bleomycin into bronchial tree through endotracheal intubation with laryngoscope. The pulmonary fanctions were measured., Results: Hedysari Radix flavonoids could normalize the pulmonary functions of rats with bleomycin-induced pulmonary fibrosis., Conclusion: Hedysari Radix flavonoids can inhibit the process of pulmonary fibrosis.

Objective: To observe and evaluate the effect of transdifferentiation of bone marrow derived stroma cells (BMSCs) into nerve cells by ultrafiltration membrane extract mixture from Angelica sinensis and Hedysarum polybotrys., Methods: The BMSCs in vitro cultured after treated by ultrafiltration membrane extract mixture from Angelica sinensis and Hedysarum polybotrys were divided into 5 groups, i.e., the blank group, the low dose group (6 g/L mixture), the high dose group (12 g/L mixture), the combination group (3 g/L mixture + 0.5 mmol/Lbeta-mercaptoethanol), and the positive control group (13-mercaptoethanol). The effects of transdifferentiation of nerve cells were observed using toluidine blue staining in each group. The differences of 5 specific neuroproteins, i.e. neuron-specific enolase (NSE), nestin, neurofilament protein (NFP), microtubule associated protein 2 (MAP2), and glial fibrillary acidic protein (GFAP) were detected using immunohistochemical technique and immunofluorescent technique respectively. The changes of the cell cycle were detected using flow cytometry (FCM)., Results: After induction BMSCs changed morphologically. The morphological features were weaker in the high and low dose groups than in the combination group and the positive group. Except the blank group, the aforesaid 5 proteins expressed positively in the rest groups. Their expression levels were highest in the positive control group (P <0.05), followed by the combination group (P <0.05). As for the cell proliferation rate detected by FCM, it was the lowest in the positive control group, followed by high dose group, low dose group, and then the combination group (all P <0.05)., Conclusions: The ultrafiltration membrane extract mixture from Angelica sinensis and Hedysarum polybotrys could effectively induce the transdifferentiation of BMSCs into nerve cells. Its inducing capacities were weaker in the positive control group, but it showed marked proliferation effects on differentiated cells. Therefore, the mixture might be a more ideal medication pathway for effectively inducing BMSCs' transdifferentiation into nerve cells, which might have higher proliferation and be used for clinical research.

Objective: To compare the contents of inorganic elements in root, stem and leaf of several species from Moghania genus and root of M. philippinensis from different habitats., Methods: Inductively Coupled Plasma Optical Emission Spectrum (ICP-OES) was used to determine twenty five inorganic elements simultaneously. The correlation analysis, principal component analysis and cluster analysis were applied for the study of inorganic elements., Results: The content of inorganic elements from different species was difference. As for M. philippinensis from different habitats, the content sequence was Guangxi > Jiangxi > Guangdong > Sichuan. There was significant correlation between different inorganic elements. The principal component analysis results showed that V, Co, K, Fe, Cr, P, Mo, Si, Na, Ca and Mg might be the characteristic elements of these species from Moghania genus. The results of cluster analysis showed that M. philippinensis and M. strobilifera could be separated and M. philippinensis from Guangxi, Jiangxi, Guangdong and Sichuan could be separated., Conclusion: This experiment provided theoretical basis for distinguish and quality assessment of species from Moghania genus and M. philippinensis from different habitats.

A reversed-phase high performance liquid chromatography coupled with tandem mass spectrometric (LC-MS/MS) method was developed for the determination of 4-chlorophenoxyacetic acid, 6-benzylaminopurine, enrofloxacin and norfloxacin residues in sprouts and source beans. The sample was extracted by acetonitrile containing 0.1% acetic acid and concentrated. The chromatographic analysis was carried out on a C18 column with methanol and 0.1% formic acid solution as the mobile phases in gradient elution program. The MS analysis was set in electrospray ionization mode and separated to two segments of positive and negative modes. The precursor ions were m/z 189.9, 226.1, 359.9 and 320.1, while the product ions for quantification were m/z 127.0, 91.2, 315.9 and 276.2 for 4-chlorophenoxyacetic acid, 6-benzylaminopurine, enrofloxacin and norfloxacin, respectively. The calibration curves showed good linearity in the range of 5 - 200 microg/L with correlation coefficients more than 0.995. The limits of detection (LODs) were 1 microg/kg and the limits of quantification (LOQs) were 5 microg/kg for the four compounds spiked in mung bean sprouts and mung beans. The recoveries of the four compounds spiked at three levels of 5.0, 10.0 and 20.0 microg/kg ranged from 70% to 91%, with the relative standard deviations (RSDs) less than 14%. The method established is accurate, sensitive, simple, and has considerable advantages in the analysis of the four kinds of illegal additive residues in sprouts and beans simultaneously.

Objective: To provide the acute toxicity data of cowpea trypsin inhibitor (CpTI) using recombinant protein purified from E. coli., Methods: Recombinant CpTI protein was expressed and purified from E. coli. Bacterial recombinant plasmid was transformed into E. coli and the transformed cells were induced with IPTG. The expressed CpTI protein was purified by hydrophobic interaction chromatography and anion exchange chromatography. Sixty mice, randomly assigned to 6 groups, were administrated 10.0, 4.64, 2.15 and 1.00 g/kg BW of CpTI or 5.00 g/kg BW of BSA control protein or sterile water respectively by oral gavage., Results: All animals survived with no significant change in body weight and food consumption throughout the study. Macroscopic necropsy examination on day 15 revealed no gross pathological lesions in any of the animals. The maximum tolerated dose (MTD) of CpTI was more than 10.0 g/kg body weight in mice., Conclusion: No toxicity of CpTI protein was found in ICR mice model.

Objective: To study the effect of PP333 treatment on seed yield,medicinal materials yield and quality of Flemingia Philippinensis., Methods: The experiment was in split plot design for PP333 in main plot and in planting density subplot., Results: The seed yield of 600 mg/L (PP333) + 60 plants/m2 (planting density) was the highest. The medicinal materials yield of 600 mg/L (PP333) + 90 plants/m2 (planting density) was the highest. The quality of 900 mg/L( PP333) + 60 plants/m2 (planting density) was the best., Conclusion: The appropriate density of PP333 and planting can improve seed yield,medicinal materials yield and quality of Flemingia Philippinensis.

Objective: To study chemical constituents contained in Desmodium caudatum., Method: The chemical compounds were separated by using such chromatographic methods as macroporous resin, Sephadex LH-20, ODS and normal phase silicagel column, and their structures were identified by spectroscopic data analysis., Result: Fifteen compounds were separated and identified as stigmasterol (1), beta-sitosterol (2), citrusinol (3), hibiscone A (4), yukovanol (5), kenusanone I (6), neophellamuretin (7), desmodol (8), erythrotriol (9), hibiscone D (10), kaempferol (11), 8-prenylquercetin (12), leachianone G (13), 5,7,4'-trihydroxy-dihydroflavonol (14), and 4H-1-benzopyran-4-one, 2-(3,4-dihydroxyphenyl) -2, 3-dihydro-3,5,7-trihydroxy-8-( 3-methyl-2-butenyl) -, (2R-trans)-(9CI) (15)., Conclusion: All of the compounds were separated from D. caudatum for the first time except compound 8.

Objective: To compare the regulating effects of Hedysari Radix and Astragali Radix alternative classic tonification prescriptions on humoral immunity in immunosuppressed mice., Methods: The immunosuppressed mouse model was induced by cyclophosphamide. The mice were administered intragastically with same dose of Hedysari Radix and Astragali Radix alternative Buzhong Yiqi Yiqi Yangxue,Yupingfeng oral liquid and Fuqi Zhihan granules for antagonistic experiments in vivo. And spleen index, HC50, CD19+B lymphocyte subgroup and content of serum IL-4 were determined after treatment., Results: Both groups of Hedyseri Radix and Astragali Radix could antagonize immunosuppressive action caused by cyclophosphamide. They both could significantly raise spleen index, HC50, CD19+ B lymphocyte subgroup and content of serum IL4 in different degree. And Yupingfeng aqueous extract of Hedysari Radix substitute Astragali Radix was better than Yupingfeng oral liquid in raising spleen index. There were no significant differences among the rest Hedysari Radix and Astragali Radix alternative groups., Conclusion: Hedysari Radix compatibility with other drugs compared with original prescription has similar role in humoral immunity regulation.

Objective: To study chemical constituents of aerial parts of Gueldenstaedtia stenophylla., Method: Chemical constituents were extracted with 95% alcohol and separated by repeated silica gel column, MCI, Sephadex LH-20 and RP C18 column chromatographic separation from aerial parts of G. stenophylla. Their structures were identified on the basis of the physiochemical properties and spectral data., Result: Twenty-two compounds were separated and identified as apigenin (1), chrysoeriol (2), farnisin (3), diosmetin (4), 4', 7-dihydroxyflavone (5), luteolin (6), 3', 4', 7-trihydroxyflavone (7), quercetin (8), m-hydroxy benzoic acid (9), trans-ferulic acid (10), isovanillic acid (11), E-beta-hydroxy-cinnamic acid (12), salicylic acid (13), trans-p-coumaric acid (14), protocatechuic acid (15), (S) -2-hydroxyphenylpropionic acid (16), esculetin (17), 7-methoxy coumarin (18), phaseic acid (19), blumenol A (20), (6S, 9R)-roseoside (21), kaempferol-7-O-alpha-L-rhamnopyranoside (22)., Conclusion: Except compounds 1, 5, 8 and 15, the rest compounds were separated from genus Gueldenstaedtia for the first time.

New method for rapid and quantitative analysis of 6-benzylaminopurine (6-BA) residues in sprout beans was studied by using FAST-SPE and portable surface-enhanced Raman spectroscopy (SERS). With homemade sprout bean samples as blank control, 6-BA solutions extracted from inserted-treatment samples showed typical characteristic Raman peaks at 1002 cm(-1) tested by SERS, and normalized 1002 cm(-1) intensities had a good linear relationship with gradient concentrations of 6-BA insert-standard solutions. The high concentration linear range was 0.5-14 microg x mL(-1), and the low one was 0. 1-2 microg x mL(-1). The quantitative detection limit was down to 0.02 mg x kg(-1) that was below the maximum allowable residues (MAL) of 0. 2 mg kg(-1) in related National Standard. The recoveries were 82.3%-95.1% and the RSD was below 5%. This method with portable equipment and simple pre-treatment showed good reproducibility, short test time-consuming and easy operating, and thus it would provide efficient and sensitive solutions for large-scale on-site and rapid detections.

Objective: To analyze the differentially expressed proteins of the synergy effect of Radix Hedysari polysaccharides (HPS)combined with chemotherapy (Cy) on S180 tumor cells., Methods: The total proteins extracted from the HPS combined with Cy treated S180 cells in tumor-bearing mice were separated by two dimentional gel electrophoresis (2-DE)and compared with those from Cy treated S180 cells using PDQuest 8.0 software. Mass spectrometry was applied to identify the differentially expressed proteins. Western blot was used to determine the differential expression of one protein., Results: Twenty-four differentially-expressed proteins in HPS group were discovered. The five differential expressed proteins among twenty-four proteins were later identified by mass spectrometry and Mascot software as heat-shock protein hsp84 (HSP90beta), apolipoprotein A, albumin, heat shock protein beta-1 (HSP27)and unnamed protein product, including one up regulated and four down regulated expressed proteins respectively. Results from Western blot manifested the same trend as from proteomics analysis., Conclusion: Proteomics technique can be used to discover target proteins associated with the synergy effect of HPS combined with Cy on S180 tumor cells, involving some important proteins related to energy metabolism, oxidative stress and apoptosis induction signal transduction.

Objective: To establish a practicable method to express and purify Cowpea Typsine Inhibitor (CpTI), which is comparable with native CpTI, for providing important materials to access the safety of genetically modified rice with CpTI., Methods: The non-fusion expression vector PET41EK-CpTI was constructed by cloning the CpTI gene into expression vector PET41EK. And the expression vector was then transformed into E. coli BL21 (DE3) and E. coli Rosetta ( DE3). Purified expression product was obtained through optimizing the induction temperature, induction time and IPTG concentration and then it was identified by western blotting, activity determination and N-terminal amino acid sequencing., Results: The purified protein was obtained under the condition of E. coli Rosetta (DE3), 20 - 25 degrees C, 130 rpm, and 0.5 mmol/L IPTG, overnight expression, 30kDa ultra-filtration and anion exchange purification. After identification, the CpTI protein with significant immunogenicity and CpTI activity was successfully obtained and its purity is up to 95%., Conclusion: The CpTI protein obtained by the non-fusion method established in this study is comparable with the native CpTI protein in terms of activity, immunogenicity, molecular weight and amino acids sequence. Moreover, the method is easy to operate and satisfied with the requirements of large-scale expression.

Objective: To study the effect of total saponins from Entada phaseoloides (TSEP) on islet morphology and skeletal muscle PI3K pathway-related protein expression of type 2 diabetic rats., Method: Type 2 diabetic rats were induced by high-fat diet and low-dose streptozotocin and then randomly divided into 5 groups, i.e. the normal control, the model group, the positive control drug (200 mg x kg(-1) metformin), the low-dose TSEP (25 mg x kg(-1)) group and the high-dose TSEP (50 mg x kg(-1)). Three weeks later, the islet morphology of rat pancreas were observed by HE staining, and protein expressions of insulin receptor substrate-1 (IRS-1), phosphatidylinositol 3-kinase (PI3K), protein tyrosine phosphatase-1B (PTP-1 B) and glucose transporter 4 (GLUT4) in rat skeletal muscle were detected by Western blot., Result: Compared with the modal group, TSEP administration groups showed relatively normal structures, clear pancreatic cells and intact capsula structures in pancreatic tissue pathological sections, with the number of pancreatic islets close to the normal control group. Meanwhile, above TSEP administration groups showed increased IRS-1, PI3K and GLUT4 protein expressions in their skeletal muscle tissues and decreased PTP-1B protein expression compared with the model group., Conclusion: TSEP has an effect on protecting pancreatic tissues of type 2 diabetic rats and intervening in abnormal expression of proteins in skeletal muscle tissues.

Objective: To study on Radix Hedyseri as substitute for Radix Astragali of Yupingfeng oral liquid on cellular immunity in immunosuppressed mice., Methods: The model of immunosuppression mice were induced by Cyclophosphamide. And the same dose of Radix Hedyseri and Radix Astragali alternative Yupingfeng oral liquid was intragastric administrated into mice; Antagonistic experiments were observed in vivo. Determined the thymus gland index, spleen index, phagocytosis of the macrophage, proliferation index of T lymphocyte, kill and wound activity, T lymphocyte subgroup, and content of IL-13 of serum., Results: Yupingfeng oral liquid and Yuping-feng aqueous extract of Radix Hedyseri substitute Radix Astragali both could significant raise thymus gland index and spleen index, and clearly increase the phagocytosis of the macrophage. They both could antagonize immunosuppressive action caused by Cyclophosphamide, which could promote T lymphocyte proliferation, kill and wound activity, quantity of T lymphocyte subgroup, and production of IL-1beta with different degree. And Yupingfeng aqueous extract of Radix Hedyseri substitute Radix Astragali increased spleen index and T lymphocyte proliferation was better than those of Yupingfeng oral liquid., Conclusion: Radix Hedyseri and Radix Saposhnikoviae, compatible with Rhizoma Atractylodis Macrocephalae compared with Yupingfeng oral liquid in cell immunity regulation has a similar role, and better in the recovery of spleen weight and T cell proliferation.

Objective: To study the chemical constituents from leaves of Uraria lacei., Method: Chemical constituents were isolated by silica gel column and Sephadex LH-20, and identified by physiochemical and spectral analyses and by comparison with the standard compounds., Result: Eleven compounds were isolated and identified as naringenin-7-0-beta-D-glucopyranside (1), (2S)-5, 7-dimethoxy-4'-hydroxyflavan (2), dalbergioidin (3), 5, 7-dihydroxy-2'-methoxy-3', 4'-methylenedioxyisoflavanone (4), apigenin (5), 5, 7-dihydroxy-2', 4'-dimethoxyisoflavanone (6), 5, 7, 2', 4'-tetrahydroxyisoflavone (7), emodin (8), saliylic acid (9), daucosterol (10), and tetracosane (11)., Conclusion: All compounds were isolated from this plant for the first time.

Objective: To establish the chromatographic fingerprints for the anti-tumor flavonoids of Caulis spatholobi (SSCE). It could used to reflect the chemical information in this part comprehensively, and identify the chemical consitituents preliminarily., Method: The HPLC-DAD analysis method was performed on the column Kromasil 100-5PHENYL (4.6 mm x 250 mm, 5 microm). The mobile phase was water (0.5% acetic acid)- methanol in gradient elution and the detection wavelength was 254 nm., Result: The chromatographic fingerprint of SSCE was established, which showed 16 characteristic peaks from 10 batches of medicinal materials. Among them, the peaks 1, 3, 4, 5, 8, 9, 10, 12, 13, and 16 were identified 3,4-dihodroxybenzoic acid, 4-Hydroxybenzoic Acid, epicatechin, puerarin, daidzein, liquiritigenin, calycosin, genistein, formononetin, and prunetin, respectively., Conclusion: The method is convenient, reproducibility and stability. It can used for quality control of the anti-tumor flavonoids of C. spatholobi (SSCE).

From the roots of Hedysarum polybotrys, ten compounds were isolated and identified as ferulic hexadecyl ester(1), ferulic(2), formononetin(3), formononetin-7-O-beta-D-glucoside(4), calycosine(5), calycosine-7-O-beta-D-glucoside(6), isoliquiritigenin(7), 7,4'-di-hydroxylflavone(8), liquiritigenin(9) and uracil(10). Compounds 2, 8, 9 and 10 were isolated from the genus Hedysarum for the first time, while compounds 6 and 7 were isolated from H. polybotrys for the first time.

Objective: To study the chemical constituents of Pithecellobium clypearia., Methods: Silica gel column chromatography and Sephadex LH-20 were used to separate and purify compounds from the EtOAc soluble fraction of Pithecellobium clypearia. 9 compounds were elucidated on the basis of physicochemical properties and spectrascopic analysis., Results: They were identified as beta-Sitosterol (1), tritriacontane (2), 5-hydroxy-3,7,3',4'-tetramethoxyflavone (3), oleanolic acid (4), 5,4'-dihydroxy-3,7,3'-trimethoxyflavone (5), alpha-amyrin (6), luteolin (7), ursolic acid (8), luteoloside (9)., Conclusion: Compounds 3 and 5 are isolated from this plant for the first time.

Six compounds were isolated from the roots of Pithecellobium lucidum by various chromatograhic techniques such as column chromatography on silica gel and Sephadex LH-20, and preparative HPLC, and their structures were elucidated as julibroside A2 (1), 3-[ (2-acetamido-2-deoxy-beta-D-glucopyranosyl) oxy] -16alpha-hydroxyolean-12-en-28-oic acid (2), galloyl acid (3), ethyl gallate (4), (+)-catechin (5), (-)-gallocatechin gallate (6) on the basis of spectrascopic data analysis. Compounds 1-6 were isolated from Pithecellobium lucidum for the first time. Compound 2 showed selective cytoxic activity against the human cell lines A2780 with an IC50 value of 1.72 micromol x L(-1).