Two new ursane triterpenoids from Agastache rugosa (Fisch. et.Mey.) O. Kuntze / 药学学报

Two new ursane triterpenoids along with twelve known compounds were isolated from 80% ethanol extract of Agastache rugosa (Fisch. et. Mey.) O. Kuntze by using silica gel column, MCI column, ODS column and HPLC. The structures of the new compounds were identified as 2α,3α-dihydroxy-24-nor-urs-4(23),12(13)-dien-28-oic acid (1) and 2α,3α-dihydroxy-24-nor-urs-4(23),12(13),20(30) -trien-28-oic acid (2) by HR-ESI-MS, NMR and ECD spectral data, named agasursacid A and agasursacid B. In addition, compounds 3, 4, 6, 8 showed anti-coxsackievirus B3 (CVB3) activities with a IC50 as 4.77, 1.59, 11.11 and 25.87 μmol·L-1, resepectively.

Seven new 3,4-dihydro-furanocoumarin derivatives from Angelica dahurica / 中草药·英文版

OBJECTIVE@#To study the chemical constituents of the roots of Angelica dahurica, a well-known Chinese herbal medicine named Baizhi in Chinese.@*METHODS@#Compounds were separated by various chromatographies, and the structures of new compounds were elucidated based on the analysis of their spectroscopic and spectrometric data (1D, 2D NMR, HRESI MS, IR, and UV). The absolute configurations of new compounds were determined by the calculated electronic circular dichroism and chemical derivatization. The inhibitory activities of all isolates against nitric oxide (NO) production were evaluated using lipopolysaccharide-activated RAW 264.7 macrophage cells.@*RESULTS@#Seven new 3,4-dihydro-furanocoumarin derivatives ( 1a/ 1b, 2a/ 2b, 3a/ 3b, 4) together with a known furanocoumarin ( 5) were isolated from the roots of A. dahurica. The new compounds included three pairs of enantiomers, (4S, 2''R)-angelicadin A ( 1a)/(4R, 2''S)-angelicadin A ( 1b), (4S, 2''S)-angelicadin A ( 2a)/(4R, 2''R)-angelicadin A ( 2b), and (4S, 2''S)-secoangelicadin A ( 3a)/(4R, 2''R)-secoangelicadin A ( 3b), together with (4R, 2''R)-secoangelicadin A methyl ester ( 4). The known xanthotoxol ( 5) inhibited the NO production with the half-maximal inhibitory concentration (IC50) value of (32.8 ± 0.8) µmol/L, but all the new compounds showed no inhibitory activities at the concentration of 100 µmol/L.@*CONCLUSION@#This is the first report of the discovery of 3,4-dihydro-furanocoumarins from A. dahurica. The results are not only meaningful for the understanding of the chemical constituents of A. dahurica, but also enrich the reservoir of natural products.

Effect of Dichloromethane Extraction Phase of Patrinia Scabiosaefolia Fisch. Stem on Proliferation and Differentiation of K562 Cells / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of dichloromethane extraction phase of ethanol extract from stem of Patrinia scabiosaefolia Fisch.(DPSS) on proliferation and differentiation of K562 cells and its related mechanism.@*METHODS@#MTT assay was used to detect the effects of DPSS at 0, 25, 50, 100 and 200 μg/ml on the proliferation of K562 cells at 24, 48 and 72 hours. Flow cytometry was used to analyze the changes of cell cycle and apoptosis at 24 and 48 hours. Wright-Giemsa staining was used to observe the morphological changes of K562 cells. The cell surface antigens CD33 and CD11b were detected by flow cytometry.@*RESULTS@#The proliferation of K562 cells treated with different concentrations of DPSS was inhibited in a time-dose dependent manner (r=-0.96). Cell cycle analysis showed that with the increase of DPSS concentration, cells in G2/M phase increased (r=0.88), and cells were blocked in G2/M phase. Flow cytometry results showed that with the apoptosis rate of K562 cells was the highest when treated with 200 μg/ml DPSS for 48 h. Morphological observation showed that the K562 cell body increased, the amount of cytoplasm increased, the ratio of nucleus to cytoplasm decreased, and the nuclear chromatin was rough after DPSS treatment. Cell differentiation antigen, CD33 and CD11b, were positively expressed after treated with DPSS.@*CONCLUSION@#DPSS can induce apoptosis through cell cycle arrest, inhibit the proliferation of K562 cells, and induce K562 cells to differentiate into monocytes, which has a potential anti-leukemia effect.

Ethanol Extract of Glycyrrhiza uralensis Fisch: Antidiarrheal Activity in Mice and Contraction Effect in Isolated Rabbit Jejunum / 中国结合医学杂志

OBJECTIVE@#To evaluate the antidiarrheal effect of ethanol extract of Glycyrrhiza uralensis Fisch root (GFR) in vivo and jejunal contraction in vitro.@*METHODS@#In vivo, 50 mice were divided into negative control, positive control (verapamil), low-, medium- and high-dose GFR (250, 500, 1,000 mg/kg) groups by a random number table, 10 mice in each group. The antidiarrheal activity was evaluated in castor oil-induced diarrhea mice model by evacuation index (EI). In vitro, the effects of GFR (0.01, 0.03, 0.1, 0.3, 1, 3, and 10 g/L) on the spontaneous contraction of isolated smooth muscle of rabbit jejunum and contraction of pretreated by Acetylcholine (ACh, 10 µmol/L) and KCl (60 mmol/L) were observed for 200 s. In addition, CaCl2 was accumulated to further study its mechanism after pretreating jejunal smooth muscle with GFR (1 and 3 g/L) or verapamil (0.03 and 0.1 µmol/L) in a Ca2+-free-high-K+ solution containing ethylene diamine tetraacetic acid (EDTA).@*RESULTS@#GFR (500 and 1,000 mg/kg) significantly reduced EI in castor oil-induced diarrhea model mice (P<0.01). Meanwhile, GFR (0.01, 0.03, 0.1, 0.3, 1, 3, and 10 g/L) inhibited the spontaneous contraction of rabbit jejunum (P<0.05 or P<0.01). Contraction of jejunums samples pretreated by ACh and KCl with 50% effective concentration (EC50) values was 1.05 (0.71-1.24), 0.34 (0.29-0.41) and 0.15 (0.11-0.20) g/L, respectively. In addition, GFR moved the concentration-effect curve of CaCl2 down to the right, showing a similar effect to verapamil.@*CONCLUSIONS@#GFR can effectively against diarrhea and inhibit intestinal contraction, and these antidiarrheal effects may be based on blocking L-type Ca2+ channels and muscarinic receptors.

Effect and molecular mechanism research of Astragalus membranaceus on inhibiting intestinal absorption of six alkaloids of Aconitum carmichaelii in spleen deficiency rats / 中草药·英文版

Objective: To investigate the effect and the mechanism of Astragalus membranaceus (Huangqi in Chinese, HQ) extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii (Fuzi in Chinese, FZ) in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier. Methods: Four-week-old male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d. Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids. Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins (TJ, including Claudin-1, Occludin and ZO-1) were measured using Western blot and real-time PCR, respectively. The location and expression of TJ protein was also investigated by the immunofluorescence method. Results: Compared with the normal group, the protein expression of MRP2, BCRP and TJ proteins in the model group were significantly down-regulated. After oral administration of HQ, the alkaloid absorption in intestinal villi was inhibited, MRP2, BCRP and TJ proteins were up-regulated, the green fluorescence staining of Claudin-1, Occludin, and ZO-1 was enhanced, and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity. Conclusion: HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components, leading to a decrease in the absorption of drug-like molecules.

A new diphenyl butanedione from Astragalus membranaceus of northern Shanxi / 药学学报

Seven compounds were isolated from Astragalus membranaceus of northern shaanxi by silica gel and Sephadex LH-20 column chromatographies. Their chemical structures were identified on the basis of their physical and chemical properties. These compounds were elucidated as astragaloside IV (1), formononetin (2), calycosin (3), 1-(4-hydroxyphenyl)-4-(2,4-hydroxyphenyl)-2-hydroxy-1,4-but anedione (4), (E)-4-methylcinnamic acid (5), quercetin (6), and uridine (7). Compound 4 is a new compound and compound 5 was isolated from the plants of Astragalus Linn. for the first time. The results of in vitro antitumor activity assay showed that compound 4 could inhibit the proliferation of A549 with IC50 values of 11.41 μmol·L-1.

Effects of different softening methods on quality of Glycyrrhizae Radix et Rhizoma / 中草药

Objective: A method was established to obtain fingerprint and determination of six components in Glycyrrhizae Radix et Rhizoma Pieces (GRRP) based on HPLC-PDA, and samples with four kinds of softening methods (showering moistening, steaming, 70 ℃ decompression steaming, 85 ℃ decompression steaming) were analyzed. Methods: The content of total flavonoids and total saponins was determined by ultraviolet spectrophotometry with liquiritin and glycyrrhizic acid as reference materials. Simultaneous determination of six components of liquiritin, ononin, isoliquiritin, glycyrrhizin, echinatin, glycyrrhizic acid was performed based on HPLC. Changes of the components content in the samples which treated by different softening methods were compared. The similarity evaluation of samples with different softening methods was carried out by the chromatographic fingerprint similarity evaluation system of traditional Chinese medicine, and cluster analysis was also carried out. Results: The results showed that the content of total flavonoids and total saponins in untreated samples was the highest, and the content of total flavonoids and total saponins in samples treated by showering moistening was the lowest. The three treatment methods of atmospheric pressure steaming, steaming decompression at 70 ℃ and steaming decompression at 85 ℃ had little effect on the samples. The content determination showed that the content of isoliquiritin was decreased significantly after softening treatment. The difference among the different softening treatment groups was not significant. The samples with different softening methods of the three batches of samples were grouped together with their raw products. Different softening methods had no significant difference in the composition of the medicinal herbs. Conclusion: The established method can quickly and accurately determine the six components, and in particular, the content of isoglycyrrhizin should be monitored. Combining production efficiency, production cost and quality evaluation, steaming is the most feasible in the production process. This study provided theoretical guidance for the large-scale production of softening, which was conducive to further standardizing the production process of GRRP.

Prescription regularity of traditional Chinese medicine for treating alcoholic liver disease based on traditional Chinese medicine inheritance support system / 中草药

Objective: To summarize the prescription rules of traditional Chinese medicine (TCM) for treating alcoholic liver disease (ALD) based on TCM inheritance support system (V2.50). Methods: The literatures about TCM prescriptions for treating ALD were collected from CNKI, Wanfang database, and VIP database. The TCM inheritance platform system was used to analyze the prescription rules of TCM in the treatment of alcoholic liver disease. Results: Statistics showed that the majority of prescriptions were used to treat alcoholic fatty liver, alcoholic hepatitis, and alcoholic cirrhosis. Through "frequency statistics" analysis, 107 prescriptions were found involving 149 flavors of TCM, with a cumulative frequency of 1 195 times. Twenty-three Chinese medicines with a frequency of ≥ 15 times were used, and the cumulative frequency was 737 times (62%). The most frequently used medicines were blood-activating and stasis-removing drugs, water-diffusing and damp-permeating drugs, tonics, heat-clearing drugs, antialcoholic poisons and qi-regulating drugs. The commonly used doses of Salvia miltiorrhiza, Poria cocos, Bupleurum chinense, Glycyrrhiza uralensis, Atractylodes macrocephala, Alismatis Rhizoma, and Curcumae Radix in the top 10 medicines ranked in the frequency of medication accorded with the prescribed doses in the Pharmacopoeia of the People's Republic of China (2015 edition), while Crataegi Fructus, Artemisiae Scopariae Herba, and Puerariae Lobatae Radix exceeded the prescribed doses. In the frequency analysis of drug pairs, the combination of S.miltiorrhiza and B. chinense was the most widely used. According to the association rules of drug combination, the correlation between Curcumae Radix and S. miltiorrhiza was the strongest, that was, the probability of S. miltiorrhiza appearing with the emergence of Curcumae Radix was 88%. From the network display chart, it was indicated that S. miltiorrhiza and P. cocos were the main herbs for treatment. Through unsupervised entropy hierarchical clustering algorithm, 14 core combinations for new clustering were extracted, and seven new prescriptions can be obtained by further clustering. Conclusion: The basic principles of TCM treatment of ALD include promoting blood circulation and removing blood stasis, removing dampness, tonifying, detoxifying alcohol, and promoting qi, and with "protecting spleen and stomach function" as its purpose, which accords with the theoretical basis of traditional Chinese medicine in treating alcoholic liver disease. Core combinations and new prescriptions provide references for clinical drug use and new drug research and development, but new prescriptions must be further evaluated with the combination of traditional Chinese medicine theory and clinical practice.

Research on rules of syndrome differentiation and medication of coronary heart disease based on data mining / 中草药

Objective: To analyze the rules of coronary heart disease syndrome differentiation based on the data mining technology. Methods: First of all, the famous medical records and prescriptions were collected and normalized to construct the database of coronary heart disease prescriptions. Then, IBM SPSS Statistics 20.0, a statistical software, was used to conduct statistics on the distribution of drug frequency, disease site syndrome frequency and disease syndrome frequency. Finally, the Apriori algorithm was applied to carry out data mining and analyze the underlying law of drug compatibility. Results: A total of 145 prescriptions were selected and analyzed, including 216 Chinese medicines, eight syndromes of disease location and 12 syndromes of disease nature. Forty-six herbs with higher frequency were found, and the top five TCM herbs were Salvia miltiorrhiza, Glycyrrhiza uralensis, Trichosanthes kirilowii, Pinellia ternata and Ligusticum chuanxiong. The main syndromes of disease location were heart, kidney, spleen and liver, while the main syndromes of disease nature were blood stasis, qi deficiency, phlegm, deficiency of yang, qi stagnation, and deficiency of yin. When the minimum support was 15% and the minimum confidence coefficient was 70%, the association rule analysis results showed that a total of 15 association rules for two-herb-combinations and 26 association rules for the trigeminy medications were obtained. The most frequency two-herb-combinations were Allium macrostemon→T. kirilowii", "Schisandra chinensis→Ophiopogon japonicus", "Curcumae Radix→S. miltiorrhiza. The most frequently used trigeminy medications were A. macrostemon + P. ternata→T. kirilowii, S. miltiorrhiza + A. macrostemon→T. kirilowii and A. macrostemon + G. uralensis→T. kirilowii. Conclusion: The syndrome differentiation and medication of TCM in the treatment of coronary heart disease mainly focus on invigorating the circulation of blood and resolving stasis, moving qi, nourishing and replenishing qi, clearing up phlegms. It is consistent with the main syndromes of the disease nature, such as blood stasis and qi deficiency, and the herbs mostly return to the heart meridian, which is consistent with the main syndromes of the disease location. The rules of syndrome differentiation and medication of based on the data mining technique have great value for clinical medication guidance and application to analyze the prescription for coronary heart disease.

Isolation and activity evaluation of endophytic fungi from Glycyrrhiza uralensis / 中草药

Objective: To isolate and purify endophytic fungus from Glycyrrhiza uralensis, and screen the specific strain with better antibacterial and antioxidant activity (DPPH• radical scavenging, total reducing power, determination of hydroxyl radical scavenging) and anti-alpha-glucosidase activity. Methods: The endophytic fungi were isolated and purified from G. uralensis by tissue cutting. N-butanol, ethyl acetate and ethanol were used to extract the fermentation liquid and mycelium. Antibacterial activity was detected by filter paper. Three methods were used to characterize antioxidant activity. A total of 36 endophytic fungi were isolated from G. uralensis by PNPG method. Results: A total of seven genera were isolated from 108 samples by concentrated fermentation liquid extraction. Trichoderma. sp was the dominant species. The experimental results showed that 43.52% of the samples had different degrees of antibacterial effect, of which 8.33% performed well; The extracts showed different levels of antioxidant activity, of which 4.63% to 10.12% showed better performance in the three methods. 99.07% of the samples had different levels of anti-α-glucosidase activity, of which 5.56% of the samples performed well. Conclusion: The strains isolated from G. uralensis have good antibacterial activity, antioxidant activity, and anti-alpha-glucosidase activity, which further study is needed.

Genetic diversity analysis on cultivar and wild population of Rehmannia glutinosa based on ISSR markers / 中草药

Objective: To study the genetic diversity and genetic relationship of cultivar and wild population of Rehmannia glutinosa and its relative species by ISSR molecular marker technique, and provide the reference for R. glutinosa germplasm protection and breeding. Methods: 106 samples including cultivar and wild population of R. glutinosa and its relative species were studied by ISSR-PCR markers. Nei's genetic diversity index (H) and other parameters of genetic information were calculated by POPGEN 32, and a cluster dendrogram of different samples was established based on the unweighted pair-group method with arithmetic mean (UPGMA) by NTSYS-pc software. Results: Seven ISSR primers generated 85 loci of which 83 loci were polymorphic. The percentage of polymorphie bands (PPB) of all samples was 97.65%. Nei's genetic diversity index (H) and Shannon's information index (I) were 0.2659 and 0.4125. The percentage of polymorphie bands (PPB) of cultivar of R. glutinosa was 30.59%. The percentage of polymorphie bands (PPB) of wild population of R.glutinosa in Henan province was 83.53%. In the cluster dendrogram, all samples were clustered into seven groups at the level of Genetic similarity coefficient (GS) 0.67. Conclusion: The results of ISSR analysis revealed that the level of genetic diversity between wild populations of R. glutinosa was higher than that within cultivar populations of R. glutinosa. The genetic diversity among wild populations of R. glutinosa in Henan province was higher than other region, which was consistent with authentic producing areas of R. glutinosa in this area. The relationships of wild population of R. glutinosa had no obvious correlation with their geographical distribution pattern.

Effects of exogenous trehalose on growth of Glycyrrhiza uralensis seedlings and total flavonoid content under salt stress / 中草药

Objective: To study the effects of exogenous trehalose on the growth and total flavonoids content of licorice (Glycyrrhiza uralensis) seedlings under NaCl stress. Methods: Using licorice seedlings as the material, the effects of NaCl stress on physiological growth, enzyme activity, ion content, osmotic regulation and total flavonoid content of licorice seedlings were studied in this experiment, Microsoft Excel 2010 was used for data processing and analysis, and SPSS19.0 statistical software was used for data analysis of variance. Results: Trehalose (10-20 mmol/L) significantly reduced the damage of NaCl to licorice seedlings, and the effect was the best when the concentration of exogenous trehalose was 15 mmol/L. Under NaCl stress, when exogenous trehalose concentration was 15 mmol/L, the growth of licorice seedlings was the most exuberant and the growth amount increased the most, and the K+ and K+/Na+ concentrations that affected the osmotic regulation of licorice seedlings increased the most, while the concentrations of Na+ and Cl- were decreased compared with those without NaCl stress. When the concentration of exogenous trehalose was 15 mmol/L, the activity of antioxidant enzymes in licorice seedlings under NaCl stress could be improved, and the content of light and chlorophyll in plant cells under NaCl stress could be increased. When the concentration of exogenous trehalose was 15 mmol/L, the contents of soluble sugar, proline and cellular regulatory substance MDA in licorice seedlings under NaCl stress could be reduced. Conclusion: The application of exogenous trehalose with appropriate concentration under NaCl stress can promote the growth of licorice seedlings and the accumulation of effective components, reduce the harm of salt damage to the growth of licorice, and enhance the growth ability of Licorice under NaCl stress.

Specific Chromatograms of Glycyrrhiza uralensis Fisch. Flavonoids in Different Growth Years by HPLC Coupled with Chemometric Analysis / 中国药学杂志

OBJECTIVE: To establish the HPLC specific chromatogram of licorice flavonoids based on chemometric analysis method, and search for differentiated components of Glycyrrhiza uralensis Fisch. cultivated for different years. METHODS: The analysis was performed on a Shiseido Capcell Pak C18 column(4.6 mm×250 mm,5 μm) by gradient elution with acetonitrile-0.1% formic acid solution as mobile phase at a flow rate of 1.0 mL•min-1. The column temperature was maintained at 40℃ and the detection wavelength was set at 280 nm. The HPLC specific chromatograms of Glycyrrhiza uralensis Fisch. cultivated for different years were further evaluated by chemometric methods including similarity analysis (SA),hierarchical clustering analysis (HCA) and principal component analysis (PCA). RESULTS: Four-years-old licorice was confirmed as the best one. Liquiritin, liquiritin apioside, liquiritigenin and isoliquiritin had significant differences, which can be used as key indicators to distinguish Glycyrrhiza uralensis Fisch. cultivated for different years. CONCLUSION: This method can provide reference for standardized cultivation and quality standard improvement of licorice.

Quality Evaluation of Glycyrrhiza uralensis Fisch. in Different Harvest Periods Based on Combinative Methods of HPLC Specific Chromatogram, Multi-Component Assay, and Chemometrics / 中国药学杂志

OBJECTIVEP: To evaluate comprehensively the quality of Glycyrrhiza uralensis Fisch. in different harvest periods by chemometric analysis based on the HPLC specific chromatogram and multi-component assay. METHODS: The similarity was analyzed with "Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica 2012". t-test, correlation analysis, clustering analysis(HCA), principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) were used for data analysis. RESULTS: Twenty-one peaks were selected as common peaks of the fingerprint. The similarity of the samples were above 0.9. The contents of liquiritin, glycyrrhizic acid, liquiritin apioside, isoliquiritin apioside, isoliquiritin, neoisoliquiritin, echinatin, and liquiritigenin were determined.There were some differences in the quality of Glycyrrhiza uralensis Fisch. in different harvest periods, with liquiritin apioside, neoisoliquiritin and echinatin as the main compounds of difference. CONCLUSION: Autumn is confirmed as the best harvesting period of Glycyrrhiza uralensis Fisch. by chemometric analysis. It is suggested that liquiritin apioside should be used as the key quality control indicator for evaluating Glycyrrhiza uralensis Fisch.in different harvest periods.

Over-expressing root-specific β-amyrin synthase gene increases glycyrrhizic acid content in hairy roots of glycyrrhiza uralensis / 中草药·英文版

Objsective: Glycyrrhizia uralensis, one of the most widely-used traditional Chinese medicines, is mainly cropped in China. However, many cultivars are less in glycyrrhizic acid than Chinese Pharmacopoeia requires. In this paper, we improved glycyrrhizic acid by regulating β-amyrin synthase gene (GuBAS). Methods: Tobacco root-specific promoter TobRB7 and GuBAS cDNA were obtained and combined with linearized pCAMBIA1305.1 to construct root-specific plant expression vector which was later transformed into Agrobacterium rhizogenes ACCC10060 by electrotransformation. The cotyledons and hypocotyls of G. uralensis were infected by the recombinant A. rhizogenes ACCC10060 to induce hairy roots. The GA content was quantified by HPLC. Results: The PCR and sequencing results both showed that three transgenic hairy root lines were obtained. The copy number of GuBAS in these transgenic hairy roots was intended by qRT-PCR to be 3, 7, and 4. GA was detected by HPLC, and the results showed that GA was present in the three transgenic hairy roots, while absent in wild hairy roots. Conclusion: Over-expressing GuBAS root-specifically in hairy roots of G. uralensis enhanced GA accumulation.

Analysis of genetic diversity of main morphological traits and chemical components in Glycyrrhiza uralensis germplasms / 中草药

Objective Based on the main morphological traits and chemical constituents of 42 Glycyrrhiza uralensis germplasm in China, the genetic diversity of them was analyzed comprehensively. Methods Twelve morphological traits and five kinds of chemical components of G. uralensis germplasm transplanted in the same place were collected. The genetic diversity index and coefficient of variation were calculated, using cluster analysis and principal component analysis for statistical analysis. Results Among the five chemical constituents, the highest genetic diversity index of liquiritin content was 2.05; The maximum coefficient of variation of isoliquiritin content was 99.50%; The content of liquiritin was moderately correlated with the content of isoliquiritin. Among 12 morphological traits, the highest genetic diversity index of plant height was 2.08, and the maximum coefficient of variation of actual fruit sequence was 37.09%. The variation of fruit type and plant type was greater than that of leaf type. Cluster analysis divided 42 germplasms into three types, and the second group had better germplasm quality. The principal component analysis reduced 17 indicators to six factors, with a cumulative contribution rate of 72.96%. Factor 6 was a factor that represents glycyrrhizic acid, liquiritin, and isoliquiritin. Conclusion The genetic diversity of the main morphological traits and chemical constituents of 42 G. uralensis germplasms is rich, and six excellent germplasms are V08, V10, V17, V34, V36, and V38.

A numerical dynamic simulation study of licorice concentration process / 中草药

Objective Taking the concentration process of liquorice extract as the research object, the dynamic simulation process of concentration of Chinese materia medica (CMM) was constructed by combining experimental analysis with theoretical simulation, which provided the model support and theoretical analysis basis for the process research and equipment development of concentration process of CMM. Methods The corresponding relationship between boiling point and saturated vapor pressure of liquorice solution was determined by dynamic method. The experimental data were fitted by thermodynamic model to obtain relevant parameters. On this basis, the simulation process of liquorice water extract concentration was constructed by using ASPEN PLUS. According to the simulation of dynamic process, the effects of heating power, feed rate, and vacuum degree on liquorice solution concentration process in an external thermal concentrator were discussed. Finally, the equations about concentration time and heating power were obtained by simulation. Results The results of parameter fitting were Aij = 1.63, Aji = 2.32, Bij = 336.38, Bji = 792.00, and Cij = 0.5. Finally, the functional equation for the concentration time and heating power was t = 2 329 c1H/c0Q. Conclusion In this study, the effects of different process parameters on the concentration process of TCM were analyzed by simulation and related theories, and a simple prediction of the concentration process was realized. It also perfected and optimized the process simulation data, filled the relevant scientific research gap, and was of great significance to industrial guidance. Firstly, the relevant experimental data was obtained by fitting the thermodynamic model with the relevant experimental data. Then, under the ideal process conditions, the influencing factors of liquorice concentration process were analyzed and discussed by dynamic simulation. It was concluded that heating power was the key factor affecting the concentration process, and the concentration time gradually decreased with the increase of heating power. However, their functional relationship was non-linear. At the same time, the functional equation can be used to roughly predict the concentration time of CMM. To a certain extent, this fills in the gap between the related theoretical research and data of chemical thermodynamics, which provides theoretical support for the process research and equipment development of the concentration process of CMM.

Preparation technology of liquiritin based on ultrafiltration-complexation extraction and back extraction technology / 中草药

Objective To establish a complexation extraction and back extraction technology for the separation and purification of liquiritin from Glycyrrhiza uralensis ultrafiltrate. Methods Taking the extraction rate of liquiritin as an index, the optimum composition of complexing extractant was first determined by uniform design, and then orthogonal experiments were used to optimize the conditions of complexing extraction. Taking the back extraction rate of liquiritin as an index, the process conditions for the back extraction of liquiritin were determined by investigating the type and concentration of back extractant. Results The complexation extraction research found that the complexing extractant should be a binary complexing extractant composed of trialkyl phosphine oxide (TRPO) and sulfonated kerosene. The optimum extraction conditions for liquiritin were as follows: TRPO-sulfonated kerosene (9∶91), pH value of G. uralensis ultrafiltrate was adjusted to 4, volume ratio of organic phase to aqueous phase was 1∶1, and average extraction rate of liquiritin reached 99.6%. The study of back extraction process showed that under the condition that the volume ratio of organic phase to back extractant was 1∶1, 17.5 mmol/L NaOH aqueous solution was the best back extractant, and the back extraction rate of liquiritin was 99.3%. Conclusion Under the optimized conditions, the liquiritin in G. uralensis ultrafiltrate can smoothly transfer from the ultrafiltrate to the complexing extractant and then to the alkaline back extractant. The total transfer rate of liquiritin is as high as 98.9%. This paper can provide a new preparation technology for the separation and purification of liquiritin.

Estimation of genome sizes of Astragalus membranaceus based on flow cytometry and K-mer analysis / 中草药

Objective: To determinate the genome size and complexity of Astragalus membranaceus by using flow cytometry (FCM) and K-mer analysis, which can lay the foundation for the screening of functional genes of A. membranaceus. Methods Lycopersicon esculentum was served as an internal reference in this study. The mixed sample of A. membranaceus cell nucleus and L. esculentum cell nucleus was stained using propidium iodide (PI). The PI fluorescence intensities of the sample were measured by FCM. The genome size of A. membranaceus was calculated by comparing the multiple relationship between the peak of DNA content in the cells of A. membranaceus and L. esculentum. The genome of A. membranaceus was sequenced by using high-throughput sequencing technologies. The genome size of A. membranaceus was calculated by K-mer analysis. The hybridity percentage, repetitive sequence, and GC of A. membranaceus were estimated by bioinformatics analysis. Results The genome size of A. membranaceus was about 1 426 Mb. For K-mer analysis, more than 95 Gb high quality data from the genome was generated. The average genome size and sequencing coverage depth of A. membranaceus was about 1 456 Mb and 39 times respectively. The genome of A. membranaceus had obvious hybridity peak by K-mer method, and the hybridity percentage as high as 2.1%. Conclusion The genome size of A. membranaceus was about 1.45 Gb and the heterozygosity is high. These data would provide a reference for the genomic research in A. membranaceus.

Cloning and related bioinformatics analysis of new cytochrome NADPH P450 reductase gene in Glycyrrhiza uralensis / 中草药

Objective To clone a new NADPH cytochrome P450 reductase gene (GuCPR) (Login number: MH401048) from Glycyrrhiza uralensis and do some bioinformatics analysis. Methods Total RNA was extracted from the roots of G. uralensis and then transcription reversed into cDNA. The GuCPR gene was obtained by screening the transcriptome database. The NCBI ORF finder was used to obtain its open reading frame and translated amino acid sequence, design primers for PCR amplification, construction of recombinant cloned plasmid. Bioinformatics analysis was used to predict the protein properties, structure and model the tertiary structure of the protein, and homologous phylogenetic tree construction was performed. Results cDNA of GuCPR gene was cloned to a total length of 2 118 bp and encoded 705 amino acid residues with a relative molecular mass of 78 450. Electricity (pI) 5.19. GuCPR protein was a non-secretory protein and did not have a signal recognition function. The results of transmembrane prediction showed that the amino acids 44—64 of the protein were transmembrane regions. Meanwhile, the subcellular localization prediction results showed that the protein was located on the endoplasmic reticulum. Conclusion A new GuCPR was cloned and its bioinformatics analysis laid the foundation for further research.