Structural characterization of chia seed polysaccharides and evaluation of its immunomodulatory and antioxidant activities.

This study aims to extract an active heteropolysaccharide Chia seed polysaccharide (CSP-A) and further purified by DEAE Sepharose Fast Flow and Sepharose CL-6B chromatographic column, characterize its structure, and evaluate its antioxidant and immunomodulatory activities. Structural analysis revealed that CSP-A was composed of d-mannose, d-glucuronic acid and d-xylose in a molar ratio of 1:3:4 with molecular weight of 1.688 × 105 Da, owning 4 sugar residues of ß-d-Manp-(1→, →4)-α-d-GlcpA-(1→, →2,4)-ß-d-Xylp-(1→, and â†’ 4)-ß-d-Manp-(1 â†’. Congo red assay and microscopic characteristics showed that CSP-A in its solution may possess a helical conformation. In vitro experiments showed that CSP-A had moderate DPPH· and OH· scavenging activities. CSP-A also enhanced the phagocytosis ability of RAW 264.7 cells and prompted the release of NO, TNF-α, IL-6 and IL-1ß from RAW 264.7 cells, which indicated CSP-A had immune regulation effect. This experiment provides scientific basis for further utilization and development of chia seeds, a kind of functional food.

Weber Texture Local Descriptor for Identification of Group-Housed Pigs.

The individual identification of group-housed pigs plays an important role in breeding process management and individual behavior analysis. Recently, livestock identification methods based on the side view or face image have strict requirements on the position and posture of livestock, which poses a challenge for the application of the monitoring scene of group-housed pigs. To address the issue above, a Weber texture local descriptor (WTLD) is proposed for the identification of group-housed pigs by extracting the local features of back hair, skin texture, spots, and so on. By calculating the differential excitation and multi-directional information of pixels, the local structure features of the main direction are fused to enhance the description ability of features. The experimental results show that the proposed WTLD achieves higher recognition rates with a lower feature dimension. This method can identify pig individuals with different positions and postures in the pig house. Without limitations on pig movement, this method can facilitate the identification of individual pigs with greater convenience and universality.

[Qualitative and quantitative analysis of major cholic acids in Suis Fellis Pulvis].

This study is to establish a qualitative method for rapid identification of bile acids in Suis Fellis Pulvis based on UHPLC-LTQ-Orbitrap-MS technology,and an HPLC-ELSD internal standard method for the quantitative determination of two glycine-conjugated BAs in Suis Fellis Pulvis.The chromatographic separation of the UHPLC-LTQ-Orbitrap-MS qualitative analysis was achieved on a Waters Acquity UPLC HSS T_3column(2.1 mm×100 mm,1.8µm),with 0.2%formic acid aqueous solution(A)-acetonitrile(B)as mobile phase ingradient elution.Electrospray ionization(ESI)source was applied and operated in negative ion mode.Quantitative analysis was performed at 30℃on a Diamonsil-C_(18)column(4.6 mm×250 mm,5µm).The mobile phase consisted of 0.2%formic acid solution and acetonitrile with gradient elution and the flow rate was 1.0 m L·min~(-1).An ELSD was used with a nitrogen flow-rate of1.4 L·min~(-1)at a drift tube temperature of 60℃and the gain was 1.A total of 14 bile acids in Suis Fellis Pulvis were characterized based on the accurate mass measurements,fragmentation patterns,chromatographic retention times,and reference materials.For the quantitative analysis method,the glycohyodeoxycholic acid and glycochenodeoxycholic acid had good linear relationship in the range of26.52-265.20 mg·L~(-1)(r=0.999 8)and 19.84-198.40 mg·L~(-1)(r=0.999 1),respectively.The average recoveries(n=6)were104.1%and 103.1%,and the RSD were 2.0%and 2.4%.The UHPLC-LTQ-Orbitrap-MS technology provides a fast and efficient qualitative analysis method for identification of bile acids in Suis Fellis Pulvis.The HPLC-ELSD internal standard method is accurate and reliable,which has reference value for the quality control of Suis Fellis Pulvis.

[Metabolic fingerprint analysis of RAW264.7 inflammatory cell model by using UPLC-Q-TOF/MS].

In order to reveal the properties of polar metabolome in inflammatory cells, we selected LPS-induced RAW264.7 inflammatory cell models as the carrier for the research of metabolic fingerprint analysis. In this study, an ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based metabolomics protocol was optimized for the extraction of polar metabolites from RAW264.7 cell line. Then orthogonal partial least squares discriminant analysis (OPLS-DA) was used to process the metabolic data, and finally, a total of 17 metabolites were selected and identified. The results showed that MeOH-CHCl3-H2O (8∶1∶1) was chosen as the optimal extraction solvent to achieve higher number of chromatographic peaks, with the best relative extraction efficiency and stability. Comparing with the normal cells, the inflammatory cells presented an abnormal metabolism in protein, carbohydrate, nucleotide and phospholipids. In this study, a UPLC-Q-TOF/MS-based metabolomics protocol for the polar metabolites from RAW264.7 cell line was developed, which may provide important information for the study of mechanism of inflammation and the anti-inflammatory drugs.

Effect of compatible herbs on the pharmacokinetics of effective components of Panax notoginseng in Fufang Xueshuantong Capsule.

Fufang Xueshuantong (FXT) is a well-known Chinese herbal formula which has been used to treat cardiovascular and ophthalmic diseases, especially diabetic retinopathy. Panax notoginseng (Burkill) F.H. Chen (PN) is the main herb of FXT, whose major bioactive constituents are ginsenosides. However, the scientific basis of the compatibility of FXT is still ambiguous. The present study investigated the scientific basis of the compatibility of FXT by comparing the pharmacokinetics of marker compounds after oral administrations of PN and FXT. A high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) method was developed for simultaneous detection of notoginsenoside R1 (NR1), ginsenoside Rg1 (GRg1), and ginsenoside Rb1 (GRb1) in rat plasma. The pharmacokinetic studies of FXT and PN were performed using the established method with the pharmacokinetic parameters being determined by non-compartmental analysis. The results showed that the pharmacokinetic parameters (maximum concentration, area under the curve (AUC0-t), clearance, and mean residence time) of NR1, GRg1, and GRb1 were significantly different after oral administration of FXT (P<0.05) compared with PN. The AUC0-t values of GRg1 and GRb1 were 1.7- and 3.4-fold greater, respectively, in FXT than in PN. The compatible herbs of FXT could prolong the retention time and increase the systemic exposure of NR1, GRg1, and GRb1 compared with PN in vivo, providing some scientific basis for the compatibility and clinical use of FXT.

Efficacy and safety of vitamin C for atrial fibrillation after cardiac surgery: A meta-analysis with trial sequential analysis of randomized controlled trials.

OBJECTIVES: Antioxidant supplement is an option in preventing postoperative atrial fibrillation (AF) after cardiac surgery. However, the benefits and adverse effects of vitamin C have not been well assessed. We aimed to systematically evaluate the efficacy and safety of vitamin C in preventing postoperative AF in adult patients after cardiac surgery. METHODS: PubMed, EMBASE, and the Cochrane library databases from inception to September 2016 were searched. Randomized controlled trials (RCTs) that evaluated the efficacy and safety of vitamin C in preventing postoperative AF in adult patients after cardiac surgery were identified. The primary outcome was the incidence of postoperative AF. Secondary outcomes included the length of intensive care unit (ICU) stay, length of hospital stay, and stroke events. RESULTS: Eight RCTs incorporating 1060 patients were included. Compared with placebo group, vitamin C treatment was associated with a substantial reduction in postoperative AF (OR, 0.47; 95% CI, 0.36-0.62; evidence rank: moderate), with no significant heterogeneity (I2 44%; P = 0.09). Trial sequential analysis showed that the cumulative Z-curve crossed the trial sequential monitoring boundary for benefit, establishing sufficient and conclusive evidence. In addition, vitamin C administration was not associated with any length of stay, including in the ICU (evidence rank: low) and hospital (evidence rank: low), respectively. CONCLUSIONS: Short-term treatment with vitamin C is safe, and may reduce the incidence of postoperative AF after cardiac surgery. Future studies as well as more high quality RCTs are still warranted to confirm the effects of different durations of vitamin C on cardiac surgery.

Intrathecal Administration of Flavopiridol Promotes Regeneration in Experimental Model of Spinal Cord Injury.

AIM: Spinal cord injury (SCI) is a serious condition of the central nervous system and it affects the quality of life and even hampers the day-to-day activity of the patient. In the current study, we investigated the efficacy of intrathecal administration of flavopiridol in an experimental animal model of SCI. The study also aimed at exploring the physiological effects of flavopiridol on neurons, astrocytes and cell cycle regulatory proteins. MATERIAL AND METHODS: In vitro scratch wound experiments were performed on female Sprague-Dawley rats (n=23). A complete hemisection to the right of T10 was made, and flavopiridol solution (200 mM, 0.8 nmol flavopiridol/animal) was delivered topically to the lesion site. Cell viability assay, in vitro scratch injury assay, cell cycle analysis using flow cytometry and behavioural assessments were performed. RESULTS: The local delivery of flavopiridol reduced cavity formation and improved regeneration of neurons with improvement in physiological performance. Flavopiridol also inhibited the migration and proliferation of astrocytes, and at the same time, promoted the survival of neurons. CONCLUSION: Intrathecal administration of flavopiridol can be a promising treatment strategy in patients with SCI and it needs to be validated in patient setting.

CTG-loaded liposomes as an approach for improving the intestinal absorption of asiaticoside in Centella Total Glucosides.

Centella Total Glucosides (CTG),obtained from Centella asiatica (L.), have been shown to possess a multitude of pharmacological activities, however, oral administeration of CTG failed to fulfill their therapeutic potentials due to the low bioavailability. In this study, the author prepared the liposomes encapsulated CTG using the ethanol injection method in order to enhance their intestinal absorption. The average particle size and the polydispersityindex(PDI) of CTG-loaded liposome in a batch are 137.0nm and 0.283, and the CTG-loaded amounts in CTG-loaded liposomes were 0.177mgmL(-1) and the zeta potential of CTG-loaded lipsomes is -21.2mV. The TEM images of CTG-loaded lipsomes showed that CTG-loaded liposomes are round and maintain high structural integrity, and their DSC thermograms indicated that CTG might be incorporated into the aqueous phase of DPPC to become more stable. The everted rat gut sac model was used to study the absorption characteristic of CTG-loaded solution in rat intestines. The cumulative absorption amount (Q) and the cumulative absorption percentage (P%) of asiaticoside in the CTG-loaded liposome was significantly higher than that in CTG (P<0.05), both the steady-state infiltration rate (Jss, µgcm(-2)s(-1)) and the permeability coefficient (Papp, cms(-1)) of asiaticoside in CTG-loaded liposomes were significantly higher than those in CTG (P<0.05), which revealed that the liposomes encapsulated CTG can promote the absorption of asiaticoside in the ileum of the rats by enhancing its transmembrane permeability. The above study will provide the experimental evidence and a reference for the development of the oral dosage forms of Centella total glucosides.

Identification of geographical origins of raw American ginseng and tablets based on stable isotope ratios.

BACKGROUND: American ginseng (Panax quinquefolius) is a medicine food homology plant, whose origin determines the medicinal and economical values. Therefore, a reliable method for the determination of its geographical origin must be established. An accurate, common and stable method to identify geographical origins of raw American ginseng and tablets was established by isotope ratio mass spectrometry (IRMS). 53 samples from 5 origins were collected and analyzed for isotope ratios of the elements C, H, O and N. The result showed that δ(2)H, δ(18)O and δ(15)N values were different among each geographical origin. The δ(2)H, δ(18)O and δ(15)N values were also used to establish discrimination model of geographical origin. According to verification test, the discrimination model of geographical origin was accurate and stable. The δ(2)H, δ(18)O and δ15N values were also found no significant difference between raw American ginseng and its tablet, so the discrimination model of geographical origin could also be used to discriminate the geographical origin of American ginseng tablet. In conclusion, the δ(2) H, δ(18) O and δ(15) N values can be used to discriminate the geographical origin of raw American ginseng and its tablet.

[A new method on investigate chemical constituents which have anti-thrombin effect by HPLC].

An in vitro anti-thrombin bioassay was developed to investigate the chemical constituents which have anti-thrombin effect from the water soluble components of Salvia miltiorrhiza. Using Chromozym TH as a probe combined with ethyl acetate Semi-micro extraction was applied to measure p-nitroaniline by HPLC. According to the results, the inactivationrate of thrombin by sodium danshensu, salvianolic acid A and salvianolic acid B under a given set of conditions were 3.06%, 77.77% and 2.35%, respectively. In the water-soluble components, salvianolic acid A has a direct inhibition of thrombin, while sodium danshensu and salvianolic acid B have no significant effect on thrombin. The method is sensitive and low consumption. It can eliminate the interference absorbed for the sample itself which can be used for screening single or multiple direct antithrombin active ingredient of herbal extract.

[DNA barcoding identification of Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix based on trnL-trnF sequences].

To optimize indices of molecular identification for authentication of Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix, four indices, including sequence similarity, specific positions, genetic distance and phylogenetic tree, were compared based on trnL-trnF sequences. Total DNA was extracted from Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix, and trL-trnF sequences were amplified and sequenced. Sequence similarity was calculated by BLAST analysis. Specific positions were compared by DNAman software. Genetic distance and phylogenetic tree were analyzed by Mega software. The results showed that the inter-specific and intra-specific similarity of P. ginseng and P. quinquefolius respectively was 100% and 99. 6%. There were four specific positions at G153A, T463A, C732G and T818C. The inter-specific genetic distance (0) of trL-trnF sequences was lower than intra-specific genetic distance (0. 004). P. ginseng can be distinguished from P. quinquefolius based on the phylogenetic tree. It is concluded that Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix can be authenticated by identification indices of sequence similarity, specific positions, genetic distance and phylogenetic tree. Index of specific positions based on trnL-trnF sequences is the most efficient index to authenticate Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix.

[Study on index components and fingerprints of crude and processed Siegesbeckia Herbs].

The change of kirenol, darutigenol and darutoside in Siegesbeckia and its first to ninth processed products were studied, and the ten fingerprints were compared, which provided the experimental basis for the study of Siegesbeckia processing tech- nology. The samples were analysed by HPLC on a SunFire-C18 column (4.6 mm x 150 mm, 5 µm) with gradient elution of acetonitrile (0.1% formic acid)-water (0.1% formic acid) at a flow rate of 1.0 mL x min(-1). Column temperaturewas 30 °C and the detected wavelength was 215, 320 nm. The calibration curves of kirenol, darutigenol and darutoside were linear in the range of 2.180-26.16, 2.900-34.80, and 1.012-6.072 mg x L(-1), respectively, and the average recoveries were 96.4%, 97.2% and 96.3% wit RSD 2.2%, 1.7% and 2.4%. This method was simple, the result was stable and had good repeatability, recovery and precision. The re- sult was the basis of the chemical contents variation in the processing of Siegesbeckia Herbs and further clarifying the effect of the changing.

Differentiation of bel-7402 human hepatocarcinoma cells induced by aqueous extracts of fresh gecko (AG) and its anti-tumor activity in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Gecko, a kind of reptile, has been widely used as a traditional Chinese medicine to treat various diseases including cancer in China for thousands of years. The aim of this study was to investigate the anti-tumor effect of AG (aqueous extracts of fresh gecko) on human hepatocellular carcinoma cell Bel-7402 in vitro and mouse H22 hepatocellular in vivo. Further to underlie the molecular mechanism of AG inducing the differentiation of Bel-7402 cells. MATERIALS AND METHODS: AG was obtained by water extracting method and qualitatively analyzed through High Performance Liquid Chromatography. The total protein concentration of AG was measured by BCA (bicinchoninic acid disodium) assay. The anti-tumor activities in vivo were analyzed through H22 (mouse hepatocellular carcinoma cell line H22) tumor xenografts mice. The cytotoxic activity of AG on Bel-7402 cells was evaluated by MTT assays. AFP (alpha fetoprotein) was detected by radioimmunoassay. ALB (albumin), ALP (alkaline phosphatase) and γ-GT (γ-glutamyl transpeptidase) were detected by biochemical methods with commercial kits. While morphological changes were observed through an inverted microscope. Moreover, the expression level of the proteins involved in MAPK (mitogen-activated protein kinase) signal pathway which was closely related to cellular differentiation was assessed by Western blot. RESULTS: AG showed obviously anti-tumor activity in vivo and anti-proliferative activity on Bel-7402 cells in vitro both dose-dependently. The number of clones of Bel-7402 cells treated with AG reduced and the cells were displaying differentiation state such as relatively bigger size and dispersed growth. The biochemical function markers of the cells were significantly changed after being treated with AG. The data showed that AFP secretion of the cells decreased 42.5%, ALB secretion increased 58.9%, the activity of ALP and γ-GT markedly decreased 67.0% and 48.5% separately when the concentration of AG was 10µl/ml, and those effects were all in a dose-dependent manner. The major original and phosphorylated signal proteins (ERK1/2 (extracellular sigal-regualted kinase 1/2), P38 (p38 MAPK) and JNK1/2 (c-Jun N-terminal kinase 1/2)) involved in MAPK signal pathway were measured and the results showed that AG activated the ERK1/2 of Bel-7402 cells. CONCLUSIONS: AG has anti-tumor activity in vivo and inhibits Bel-7402 cell proliferation in vitro through inducing cell differentiation, and the mechanism involves the activation of ERK1/2.

[Characteristic of sample banks isolated from EDTA-blood by sedimentation method].

OBJECTIVE: To assess the characteristics of establishing the different sample banks of plasma, leukocytes and DNA by sedimentation method of isolating from ethylene diamine tetraacetic acid(EDTA)-blood and to clarify the sedimentation method of leukocyte isolation and plasma volume by comparative data and recommended procedures for applicability. METHODS: In the study, 29 EDTA-bloods were obtained, the total amounts of leukocytes and the percentage of neutrophile granulocytes, and lymphocytes in the EDTA-blood detected as a control group and then assigned equally into 4 EP tubes with 1 mL EDTA-blood per tube as 4 test groups, then the 4 tubes were placed with the EDTA-blood at room temperature and the plasma layers were isolated at 0.5, 1, 2 and 3 h, receptively. The total amount of leukocytes and the percentage of neutrophile granulocytes, and lymphocytes were detected by automated hematology analyzer at the clinical laboratory. The volume of the plasma was also measured at the same time. RESULTS: The plasma volume at 0.5 h [(241.72 ± 101.52)µL] was substantially lower than those at 1 h[(317.24 ± 97.50)µL], at 2 h[(371.03 ± 91.66)µL], and at 3 h [(408.97 ± 97.43)µL] , P < 0.05. The plasma volume at 1 h was substantially lower than those at 2 h and 3 h (P < 0.05). The total amount of leukocytes in the plasma layer at 0.5 h (2.50 × 10(6) ± 1.48 × 10(6)) group was substantially higher than the amount of 2 or 3 h groups respectively (1.47 × 10(6) ± 7.19 × 105,1.21 × 10(6) ± 7.41 × 105), P < 0.05. Significant difference was not found between 0.5 h group and 1 h group (2.29 × 10(6)± 1.17 × 10(6)), P > 0.05. The total amount of leukocytes in the plasma layer in 1 h group was substantially higher than that in 2 h and 3 h groups (P < 0.05). There was no significant difference between 3 h group and 2 h group (P > 0.05). The total amount of leukocytes in the plasma layer of the 4 test groups was substantially lower than that in the control group (P < 0.05). The percentage of neutrophile granulocytes (54.14% ± 11.65%) in the plasma layer in 0.5 h group was substantially higher than those in 1 h, 2 h and 3 h groups (46.66% ± 12.70%,39.17% ± 12.33%,43.25% ± 14.54%), P < 0.05, respectively, which was the substantially lower than that in the control group (60.53% ± 8.46%), P < 0.05. The average value of the percentage of neutrophile granulocytes in the plasma layer in 1 h group was substantially higher than that in 2 h group (P < 0.05). There was no significant different between 3 h group and both 1 h, 2 h groups (P > 0.05). The mean percentage of lymphocytes in the plasma layer in 0.5 h group (35.09% ± 10.84%) was substantially lower than those in the plasma layer in 1 h, 2 h and 3 h groups, respectively ( 41.48% ± 12.20%, 47.96% ± 12.27%, 45.50% ± 13.71%), which was significant higher than that in the control group(30.98% ± 7.33%), P < 0.05. The average value of the percentage of lymphocytes in the plasma layer in 1 h group was substantially higher than those in the control group and 0.5 h group, but was substantially lower than those in 2 h and 3 h groups (P < 0.05). The average value of percentage of lymphocytes in the plasma layer in 2 h group was substantially higher than those in the control group, 0.5 h and 1 h groups (P < 0.05). There was no significant difference between 2 h and 3 h groups (P > 0.05). CONCLUSION: The best period of time in obtaining leukocytes is 0.5-1 h sedimentation of EDTA-blood. Both the plasma layer and leukocytes can be separated and obtained at the same time from the same sample by the sedimentation method of EDTA-blood. The sedimentation of EDTA-blood has the least interference of both chemical and physical factors, as well as a ready operation, which can establish the plasma, leukocytes and DNA sample banks for various aspects of research.

[A new herbs traceability method based on DNA barcoding-origin-morphology analysis--an example from an adulterant of 'Heiguogouqi'].

The fruit of Lycium ruthenicum is a common folk medicine in China. Now it is popular for its antioxidative effect and other medical functions. The adulterants of the herb confuse consumers. In order to identify a new adulterant of L. ruthenicum, a research was performed based on NCBI Nucleotide Database ITS Sequence, combined analysis of the origin and morphology of the adulterant to traceable varieties. Total genomic DNA was isolated from the materials, and nuclear DNA ITS sequences were amplified and sequenced; DNA fragments were collated and matched by using ContingExpress. Similarity identification of BLAST analysis was performed. Besides, the distribution of plant origin and morphology were considered to further identification and verification. Families and genera were identified by molecular identification method. The adulterant was identified as plant belonging to Berberis. Origin analysis narrowed the range of sample identification. Seven different kinds of plants in Berberis were potential sources of the sample. Adulterants variety was traced by morphological analysis. The united molecular identification-origin-morphology research proves to be a preceding way to medical herbs traceability with time-saving and economic advantages and the results showed the new adulterant of L. ruthenicum was B. kaschgarica. The main differences between B. kaschgarica and L. ruthenicum are as follows: in terms of the traits, the surface of B. kaschgarica is smooth and crispy, and that of L. ruthenicum is shrinkage, solid and hard. In microscopic characteristics, epicarp cells of B. aschgarica thickening like a string of beads, stone cells as the rectangle, and the stone cell walls of L. ruthenicum is wavy, obvious grain layer. In molecular sequences, the length of ITS sequence of B. kaschgarica is 606 bp, L. ruthenicum is 654 bp, the similarity of the two sequences is 53.32%.

[Quality survey of different species of clematidis radix et rhizoma].

Quality survey of different species of Clematidis Radix et Rhizoma was made by determining the content of hederagenin and oleanolic acid from Clematidis Radix et Rhizoma. The result showed that only a few samples of Clematis chinensis met the quality standard for Clematidis Radix et Rhizoma in Chinese Pharmacopoeia 2010 Edition.

[Comparison research on quality of cultivated and wild Anemarrhena Rhizome from Yi County].

OBJECTIVE: To compare the quality of cultivated and wild Anemarrhena Rhizome from Yi County (Xiling Zhimu) based on contents analysis of active constituents. METHOD: Samples of cultivated Anemarrhena Rhizome from most townships of Yi County were analyzed and compared with wild ones. Six indexes belonged to three kinds active constituents of saponin, flavornoid and polysaccharide were adopted. HPLC-ELSD method with cholesterol as internal standard was adopted to determine the content of sarsasapongenin. HPLC-ELSD method was used to simultaneously determine the contents of anemasaponin C and Anemasaponin A III. Contents of neomangiferin and mangiferin were determined by HPLC-UV method. Total polysaccharide was determined by phenol sulfate method. RESULT: The mean content of sarsasapongenin in cultivated Anemarrhena Rhizome samples is slightly lower than the wild. The mean contents of anemasaponin C and Anemasaponin A III in cultivated Anemarrhena Rhizome samples are higher than the wild. There is no notable difference of these three index between the cultivated and the wild. The cultivated Anemarrhena Rhizome samples have a lower content of neomangiferin and a higher content of mangiferin than the wild. While the total content of these two flavonoids have no notable difference. The cultivated Anemarrhena Rhizome samples have a higher content of total polysaccharide than the wild samples. CONCLUSION: Contents of active constituents in cultivated Anemarrhena Rhizome from Yi County (Xiling Zhimu) are not notably different with the wild Anemarrhena Rhizome. They have similar good quality as the wild ones.

[Simultaneous determination of two saponnins in anemarrhenae rhizoma by HPLC-ELSD].

OBJECTIVE: To establish an HPLC-ELSD method for determination of Anemarsaponin C and Anemarsaponin A III in Anemarrhenae Rhizoma. METHOD: Kromasil C18 column(4.6 mm x 250 mm, 5 microm) was used as stationary phase. Mobile phase was methanol-water gradient with the flow rate of 1 mL x min(-1); the temperature of the drift tube and evaporation was 50 degrees C and 70 degrees C respectively. The gas pressure was 1.03 x 10(5) Pa. RESULT: There are good linearity in the range 0.310-3.10 microg of anemarsaponin C (lgA = 1.254 2lgM + 5.734 7, r = 0.999 5) and in the range 0.323-3.23 microg (lgA = 1.328 41gM + 5. 937, r = 0.999 6) of anemarsaponin A III. The average recovery of anemarsaponin C and anemarsaponin A III was 98.1% with RSD 2.1% and 97.3% with RSD 1.5% (n = 6) respectively. CONCLUSION: The method is rapid and accurate. It is suitable for quality control of Anemarrhenae Rhizoma. The result of determination reveals that the quality of Anemarrhenae Rhizoma from different places of north China are of notable difference.

[Difference of hypaconitine concentration in serum between cold-deficiency and normal mice].

OBJECTIVE: To investigate the difference of hypaconitine concentration in serum between normal and cold-deficiency mice after administration of aconite decoction. To analyze how the toxic dose of aconite decoction correlate to the metabolic environment. METHOD: Prepared cold-deficiency mice model, treated normal and cold-deficiency mice with aconite decoction for 14 days continuously, and then detected hypaconitine concentration in serum by HPLC along with survival ratio of mice on the first, seventh and fourteenth day. RESULT: After administration of aconite decoction for 14 days, the hypaconitine concentration in serum of cold-deficiency mice is close to that in normal mice. It showed aconite decoction has the ability of regulating metabolism environment, the hypaconitine concentration in serum of normal mice was higher on the seventh and fourteenth day than that on first day. It showed that aconite decoction can disturb metabolism environment of normal mice. It was also been observed that the range of variation of hypaconitine concentration in cold-deficiency mice was minor than that in normal mice during the fourteen days' administration. CONCLUSION: The difference of serum concentration in normal and cold-deficiency mice showed that there were different metabolic environments in two mice models, and the metabolic environment changed during administration. These results showed that the different toxic doses of aconite decoction were partially due to the different metabolic environments.

[Two new sesquiterpene lactones from the pericarp of Illicium macranthum].

Silica gel column chromatography was used for the isolation and purification of the chemical constituents of the pericarp of Illicium macranthum. From dichloromethane-EtOAc (1:1) fraction and EtOAc fraction of the methanol extracts, eleven compounds were identified on the basis of chemical and spectral data. Two new compounds were elucidated to be 6-deoxyneomajucin (1) and 2-oxo-6-deoxyneomajucin (2), along with nine known compounds 6-deoxypseudoanisatin (3), pseudoanisatin (4), anisatin (5), pseudomajucin (6), protocatecheuic acid (7), shikimic acid (8), shikimic acid methylester (9), beta-sitosterol (10) and daucosterol (11). Compounds 1 and 2 are new majucin-type sesquiterpene lactones.