Effects of Ligusticum Chuanxiong extract on MPP+-induced SH-SY5Y cells in Parkinson’s syndrome model via miR-23a-3p/SNCA axis / 西安交通大学学报(医学版)

【Objective】 To explore the effects of Ligusticum Chuanxiong extract on MPP+-induced SH-SY5Y cell damage and Parkinson’s syndrome. 【Methods】 1-methyl-4phenylpyridine ion (MPP+) interfered with SH-SY5Y to establish a cell model of elderly Parkinson’s syndrome (SH-SY5Y-MPP+). After intervention with Ligusticum Chuanxiong extract, cell proliferation and apoptosis as well as miR-23a-3p and SNCA expressions were detected. In addition, the changes of SH-SY5Y-MPP+ after regulating the expression of miR-23a-3p and SNCA were observed, and the relationship between miR-23a-3p and SNCA was verified by dual luciferase reporter. 【Results】 The cell proliferation capacity of SH-SY5y-MPP+ was significantly lower than that of SH-SY5Y, while the apoptosis rate was higher than that of SH-SY5Y (P<0.05). Under the intervention of Ligusticum Chuanxiong extract, the proliferation ability of SH-SY5Y-MPP+, and Bcl-2 and SNCA protein increased, the apoptosis rate, miR-23a-3p, and Bax proteins decreased (P<0.05). Both silencing miR-23a-3p and increasing SNCA could promote the proliferation of SH-SY5Y-MPP+ and inhibit apoptosis, while increasing miR-23a-3p and silencing SNCA were the opposite (P<0.05). The online target gene prediction website found that miR-23a-3p and SNCA had complementary sites that could bind, and the dual luciferase reporter enzyme showed that the firefly activity of SNCA-wt was significantly inhibited after the miR-23a-3p mimic sequence was transfected (P<0.05). After increasing miR-23a-3p, the expression of SNCA protein in SH-SY5Y-MPP+ decreased, while silencing miR-23a-3p was the opposite (P<0.05). Rescue experiments showed that the intervention effect of Ligusticum Chuanxiong extract on SH-SY5Y-MPP+ was completely reversed by increasing miR-23a-3p or silent SNCA (P>0.05); the effect of increasing miR-23a-3p on SH-SY5Y-MPP+ increased SNCA reversion (P>0.05). 【Conclusion】 Ligusticum Chuanxiong extract can affect the biological behavior changes of SH-SY5Y induced by MPP+ by regulating the miR-23a-3p/SNCA axis, which may be a new direction for the treatment of elderly Parkinson’s syndrome in the future.

Nanocrystals self-stabilized Pickering emulsion loaded with active components of Tongmai prescription: preparation, characterization and evaluation by Caco-2 cell model / 药学学报

It is of great significance to apply the nanocrystals self-stabilized Pickering emulsion (NSSPE) to traditional Chinese medicine (TCM) compounds, and to study the effect of NSSPE on the oral absorption of various components with different solubility and permeability. In the study, NSSPE of Tongmai prescription was prepared by the high pressure homogenization method with nanocrystals of main active components (puerarin, ferulic acid, salvianolic acid B and tanshinone IIA) of Tongmai prescription as solid particle stabilizers and a mixture of Ligusticum chuanxiong essential oil and Labrafil M 1944 CS as oil phase. The NSSPE had better physical stability than nanocrystals suspension and blank emulsion. The adsorption of nanocrystals on the surface of oil droplets was confirmed by scanning electron microscopy and fluorescence microscopy. The surface adsorption rates of puerarin, ferulic acid, salvianolic acid B and tanshinone ⅡA in NSSPE were 15.40% ± 3.19%, 15.39% ± 5.07%, 10.97% ± 3.70% and 31.51% ± 1.60%, respectively. When solid active components were prepared into nanocrystals suspension, the cellular uptake and transport across Caco-2 cells were increased significantly for puerarin and tanshinone IIA. The uptake rates of ferulic acid, ligustilide and tanshinone IIA in NSSPE were further increased compared with the physical mixture of nanocrystals suspension and oil, and the transports of ligustilide and tanshinone IIA were also significantly improved. The main absorption mechanisms of NSSPE were passive diffusion and caveolin-mediated endocytosis, which were determined mainly by the microstructure of NSSPE. In conclusion, NSSPE could be applied to complicated TCM. The "micro" and "nano" synergistic microstructure with drug nanocrystals adsorbed on the surface of micron-sized oil droplets could not only improve the physical stability of NSSPE, but also promote the absorption of various components in NSSPE, which made NSSPE a promising oral drug delivery system for TCM.

Planning of ecologically suitable areas for Ligusticum chuanxiong under background of soil cadmium pollution / 中国中药杂志

Chuanxiong Rhizoma is a traditional Chinese medicinal material mainly produced in Sichuan and Chongqing of China. In recent years, the cadmium content in Chuanxiong Rhizoma produced in most of the genuine producing areas has exceeded the standard, which makes Chuanxiong Rhizoma difficult to be exported. To solve the problem of excessive cadmium content in soil, this study employed the MaxEnt model to simulate the potential geographic distribution of Ligusticum chuanxiong and evaluate important environmental factors, and re-plan its ecologically suitable areas based on the mineral distribution characteristics and soil cadmium pollution status. The results showed that the places suitable for L. chuanxiong growing covered an area of 335 523.69 km~2, mainly in central and eastern Sichuan, southern Shaanxi and most parts of Chongqing. Among them, the highly suitable areas of L. chuanxiong were mainly concentrated in Chengdu, Ya'an, Deyang, and Mianyang. Solar radiation, annual precipitation, and annual range of temperature were evaluated as important variables affecting the distribution of L. chuanxiong, with the contribution rates of 62.3%, 13.3%, and 6.8%, respectively. In addition, Qionglai county, Chongqing county, Mianyang city(Youxian district and Fucheng district), Qingchuan county, and Xinjin county were classified into the first-class ecologically suitable zone, covering a total area of 2 768.87 km~2. The se-cond-class ecologically suitable zone was even wider, involving such counties as Tongjiang county, Renshou county, Jianyang county, and Nanjiang county, and the total area reached 43 616.92 km~2. The re-planning of the ecologically suitable areas for L. chuanxiong has provided strong data support for the cultivation and resource development of L. chuanxiong and also new ideas for solving the problem of excessive cadmium content in L. chuanxiong.

Study on oral absorption mechanisms of puerarin in nanocrystals self-stabilized pickering emulsion / 中国中药杂志

Nanocrystals self-stabilized Pickering emulsion(NSSPE) is a new kind of emulsion where only nanocrystals of poorly soluble drugs are used as stabilizers. Our previous study showed that NSSPE with Ligusticum chuanxiong oil as the main oil phase can significantly promote oral absorption of puerarin. The present study aimed to explore its absorption mechanism in oral administration. The in vitro dissolution test was carried out to study the effect of NSSPE on release of puerarin. The effects and mechanism of NSSPE on uptake and transport of puerarin across Caco-2 cell were investigated. The results showed that the drug release rate of NSSPE was similar to that of nanocrystals, with their cumulative dissolution of puerarin not affected by pH of releasing mediums, both significantly higher than that of crude material. The uptake of puerarin in NSSPE was concentration-dependent and significantly higher than that of solution or surfactant stabilized emulsion. Genistein and indomethacin, inhibitors of lipid rafts/caveolin, could significantly reduce the uptake of puerarin in NSSPE. Compared with solution, NSSPE and surfactants stabilized emulsion obviously increased transport rate K_a and apparent permeability coefficient P_(app) of puerarin in AP → BL direction, but there was no significant difference in BL → AP direction. It could be inferred that there were both passive and active transport mechanisms, as well as lipid raft/caveolin mediated endocytosis for absorption of NSSPE. The promoted oral absorption of puerarin in NSSPE was mainly related to the existing nanocrystal form which could promote dissolution, puerarin as well as Ligusticum chuanxiong oil which could promote drug transmembrane transport and inhibit drug efflux. It is the unique structure and composition of the compound NSSPE that promoted the oral absorption of puerarin.

Mechanisms exploration of Angelicae Sinensis Radix and Ligusticum Chuanxiong Rhizoma herb-pair for liver fibrosis prevention based on network pharmacology and experimental pharmacologylogy / 中国天然药物

Angelicae Sinensis Radix (Danggui) and Ligusticum Chuanxiong Rhizoma (Chuan Xiong) herb-pair (DC) have been frequently used in Traditional Chinese medicine (TCM) prescriptions for hundreds of years to prevent vascular diseases and alleviate pain. However, the mechanism of DC herb-pair in the prevention of liver fibrosis development was still unclear. In the present study, the effects and mechanisms of DC herb-pair on liver fibrosis were examined using network pharmacology and mouse fibrotic model. Based on the network pharmacological analysis of 13 bioactive ingredients found in DC, a total of 46 targets and 71 pathways related to anti-fibrosis effects were obtained, which was associated with mitogen-activated protein kinase (MAPK) signal pathway, hepatic inflammation and fibrotic response. Furthermore, this hypothesis was verified using carbon tetrachloride (CCl

Application of Bioaccessibility in Health Risk Assessment of Cd in Traditional Chinese Medicines / 中国药学杂志

OBJECTIVE: To investigate and apply the bioaccessibility of Cd in traditional Chinese medicines (TCMs) to human health risk assessment, and provide reference for the formulation of limit standards. METHODS: The contents of Cd in Coptis chinensis Franch., Ligusticum chuanxiong Hort., Lindera aggregate (Sims) Kosterm., and Curcuma kwangsiensis S.G. Lee et C.F. Liang were determined by inductively coupled plasma mass spectrometry (ICP-MS). The bioaccessibility of Cd in TCMs was investigated by in vitro physiologically based extraction test(PBET) digestion method. And the target hazard quotient (THQ) was applied to assess the health risks of Cd contents. RESULTS: The bioaccessibility of Cd for Coptis chinensis Franch., Ligusticum chuanxiong Hort., Lindera aggregate (Sims) Kosterm., and Curcuma kwangsiensis S.G. Lee et C.F. Liang was 49.0%, 40.0%, 44.9% and 65.7%, respectively. The results of risk assessment showed that before extraction, THQ values of Cd for Coptis chinensis Franch., Ligusticum chuanxiong Hort., Lindera aggregate (Sims) Kosterm., and Curcuma kwangsiensis S.G. Lee et C.F. Liang were 1.895, 1.045, 1.446 and 1.653, respectively; after extraction, THQ values of Cd for the four types of TCMs were 0.929, 0.418, 0.649 and 1.077, respectively. CONCLUSION: Cd in the four types of TCMs cannot be completely absorbed in the artificial gastrointestinal phase, and the risks of Cd are reduced after extraction. Except THQ for Curcuma kwangsiensis S.G. Lee et C.F. Liang, the THQ values are below the value of 1, indicating very low health risks for the other three types of TCMs after extraction. In this study, the bioaccessibility of heavy metals in TCMs is applied to the health risk assessment model in order to provide a scientific basis for objectively evaluating the health risks of heavy metals in TCMs and formulating a scientific and reasonable heavy metal limit standards.

Three butylphthalide derivatives from the Rhizome of Ligusticum chuanxiong / 药学学报

Three butylphthalide derivatives were isolated from the Rhizome of Ligusticum chuanxiong using a series of isolation and purification approaches including macroporous resin, ODS-A column, Sephadex LH-20 and preparative HPLC. These structures were elucidated based on extensive spectroscopic data (UV, IR, HR-ESI-MS and NMR) and identified as (3Z,3aE)-(6R,7R,2'S)-6-hydroxy-7-(2-carboxyl-2-hydroxyethylthio)-3-(2-hydroxybutylidene)-4,5,6,7-tetrahydro-phthalide (1), (3Z,3aZ)-3-butylidene-6,7-dihydroxy-4,5,6,7-tetrahydro-phthalide 7-O-α-D-glucopyranosyl-(1→2)-β-D-fructo-furanoside (2) and 3-(3-β-D-glucopyranosyloxy-butylidene)-7-hydroxy-phthalide (3).

Development of germplasm identification markers and phylogenetics analysis of Ligusticum chuanxiong / 中草药

Objective: In this study, InDel markers were developed based on the high frequency Insertion/Deletion region of chloroplast genome of Ligusticum. Germplasm identification and phylogenetic development of Ligusticum chuanxiong and its common adulterants were studied with universal barcode. Methods: The 26 samples of L. chuanxiong and its common adulterants were amplified and sequenced by eight DNA universal barcodes: ycf1, matK, ITS2, rpoC1, rbcL, rpoB, trnK, and psbA-trnH. Genetic distance statistics, barcoding gap and phylogenetic tree analysis methods were used to study the phylogenetic relationship and phylogeny of L. chuanxiong. At the same time, the evolutionary tree was constructed to study molecular identification of L. chuanxiong and its common adulterants. Results: The results showed that rbcL conserved site was the highest (97.32%) with the highest GC content (44.9%). The rbcL+rpoB fragment had the smallest average intraspecific genetic distance (0.002 5). The psbA-trnH sequence fragment had the largest average interspecific genetic distance (0.429 2). The trnK and rbcL+rpoB sequence had the highest interspecific genetic distance. The overlap of the "barcoding gap" region of psbA-trnH was the least. The species of L. chuanxiong and other adulterated species were not accurately identified by the eight pairs of DNA barcodes. The cluster analysis of 24 InDel markers could accurately identify genuine L. chuanxiong and classify the species of L. chuanxiong and its adulterants into four categories, one of which was genuine L. chuanxiong collected from Sichuan. Conclusion: The ability of InDel markers to identify authentic L. chuanxiong and its common adulterants was higher than that of common barcode. According to the above studies, it is found that it is impossible to distinguish L. chuanxiong and its common adulterants by the traditional DNA barcodes because of the large difference in genetic components. The newly developed InDel molecular markers can effectively identify L. chuanxiong and its commonly used adulterants, and provide an effective method for the genuineness of L. chuanxiong at molecular level.

Bioassay of anti-gel bioactivity of Chuanxiong Rhizoma and related Chinese patent medicines / 中草药

Objective: Owing to fibrinogen dosage, this study is to develop a method for the bioassay of anti-gel bioactivity of Chuanxiong Rhizoma in vitro, and evaluate the antithrombotic effect of Chuanxiong Rhizoma and its related Chinese patent medicines. Methods: Chuanxiong Rhizoma powder was extracted in ethanol and water, respectively. The mixed extract was used as sample to quantify the level of fibrinogen when the stable fibrin gel was formed in vitro. The fibrinogen dosage was chosen as biomarker for anti-gel bioactivity. Sodium ferulate was chosen as reference. The anti-gel bioactivity of Chuanxiong Rhizoma was calculated according to the direct measurement belong to the bioassay statistical method in the Chinese Pharmacopoeia 2015 (Vol. 4). Moreover, the amounts of anti-gel bioactivity were quantified in nine Chuanxiong Rhizoma samples including Chuanxiong Rhizoma raw materials, decoction pieces, and related Chinese patent medicines to evaluate the applicability for this developed method. Results: Both sodium ferulate and the extract of Chuanxiong Rhizoma showed significant anti-gel bioactivity (P < 0.01). The RSD for the amounts of anti-gel bioactivity was 4.00% (n = 6) in six replicated tests with the confidence limit rate of 7.82% (n = 6). The amounts of anti-gel bioactivity were significant difference among the various types of Chuanxiong Rhizoma samples, i.e. 0.72-1.14 U/g for five Chuanxiong Rhizoma raw materials, 0.68 and 1.32 U/g for Chuanxiong Rhizoma decoction pieces and processed slice with yellow wine, 2.56 and 2.51 U/g for Tongmai Granules and Xuefu Zhuyu Tablets. Conclusion: The developed method can accurately quantify the level of anti-gel bioactivity in Chuanxiong Rhizoma raw materials, decoction pieces and related Chinese patent medicines to assess their antithrombotic effect.

Discovery of phthalides with vasodilating activity in stems and leaves of Ligusticum chuanxiong / 中草药

Objective: To study the phthalides with vasodilating activity of alcohol extracts from stems and leaves of Ligusticum chuanxiong. Methods: Compounds were isolated by extensive chromatographic techniques including MCI, silica gel, Sephadex LH-20, ODS, and semi-preparative HPLC. The vasodilating effect of phthalide and dimer phthalide on thoracic aortic ring in vitro was investigated. Results: Ten phthalides were isolated and identified, and their structures were elucidated as Z-3-butylidenephthalide (1), senkyunolide-E (2), Z-senkyunolide-H (3), (3Z,7β)-3-butenyl-7-hydroxy-4,5,6,7-tetrahydrophthalide (4), neoligustilide (5); Z,Z’-3.3’a,7.7’a-diligustilide (6), 3Z,3Z’-6,8’,7,3’-diligustilide (7), Z-tokinolide A (8), (3’Z)-(3S,8R,3a’S,6’R)-4,5-dehydro-3.3’a,8.6’-diligustilide (9), and (3Z)-(3aR,6S,3’R,8’S)-3a.8’,6.3’-diligustilide (10). Compound 1, 6-9 significantly reduced the strain of thoracic aortic ring in KCl preconstricted rats. The diastolic rate of compounds 6-8 were 60%, 52% and 70% at the highest concentrations (12 μmol/L), respectively. The EC50 was 9.46, 11.86 and 8.73 μmol/L, respectively. Firstly isolated from the stems and leaves of L. chuanxiong. Compounds 6, 8 are firstly isolated from L. chuanxiong. The test compounds could dilate blood vessels, and the activity of dimer phthalide was better than that of phthalide.

Anti-Parkinson mechanism of Foshou Powder based on factorial design / 中草药

Objective: To explore the effects and potential mechanism of Foshou Powder on Parkinson’s disease. Methods: Reserpine was injected in KM mice to establish Parkinson’s disease model, and hypothermia and akinesia were measured to evaluate the effects of volatile oil of Angelica sinensis, Ligusticum chuanxiong and combination. The human monoamine oxidase (hMAO) activity and inhibition mode were measured by fluorescence spectrophotometry in vitro with kynuramine as the common substrate of human monoamine oxidase A (hMAO-A) and human monoamine oxidase B (hMAO-B). On this basis, the 2 × 4 factorial design was applied to research the interaction between Angelica sinensis volatile oil and Ligusticum chuanxiong volatile oil. Results: The antagonistic experiments of reserpine showed that the application of Angelica sinensis volatile oil and Ligusticum chuanxiong volatile oil significantly improved hypothermia and akinesia. Enzyme activity test revealed that volatile oil of Angelica sinensis and Ligusticum chuanxiong had competitive inhibitory effects on hMAO-A and hMAO-B, and the combination of the two substances had significant synergistic effect on the activity of both hMAO. Conclusion: Angelica sinensis and Ligusticum chuanxiong in Foshou Powder have significant synergistic effects in the prevention and treatment of Parkinson’s disease, which may be related to the inhibition of MAO activity.

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.

Comparison on Volatile Oil Content and Chemical Constituents Among Different Parts of Ligusticum chuanxiong and Pieces with Different Processing Methods / 中国实验方剂学杂志

Objective::To compare the difference of the content of volatile oil and the total relative percentage of phthalein compounds in volatile oil among different parts about Ligusticum chuanxiong, or among different decoction pieces with different processing methods. Method::Steam distillation was used(in the extraction of volatile oil.The chemical constituents of the volatile oil were identified by GC-MS analysis, and relative content of each component was determined by normalization method. Result::The contents of volatile oil in different parts were obviously different, and the order of the contents from high to low was rhizome(1.12%)>fibrous root(0.75%)>aerial part(0.41%). The GC-MS analysis similar compounds find in the three different volatile oils, and the order of total relative percentages of phthalein compounds from high to low was roots(83.29%)>rhizomes(44.5%)>aerial part(39.95%). On the other hand, the volatile oil content of three different Chuanxiong Rhizoma pieces with different processing methods was C(0.87%)>A(0.75%)>B(0.7%). The total relative percentages of phthalein compounds analyzed by GC-MS was C(79.14%)>A(73.09%)>B(67.29%). Conclusion::The content of phthalein compounds in the volatile oil of fibrous root was higher than that of rhizome, thus it can be appropriately used.The volatile oil content and chemical composition of different Chuanxiong Rhizoma pieces were significantly different.The content of volatile oil and phthalein compounds in fresh-cut Chuanxiong Rhizoma pieces were the most high, thus fresh-cutting can be used as a new processing method for Chuanxiong Rhizoma pieces.

Research on Natural Boundary of Traditional Geo-authentic Origin of Ligusticum chuanxiong via Soil Elements Characteristics Along Same Longitude / 中国实验方剂学杂志

Objective:To find the natural environmental boundary of the origin of the authentic Ligusticum chuanxiong by analyzing the distribution characteristics of soil elements in the traditional geo-authentic area and the southern continuous expanding areas along the same longitude. Method:The contents of 24 elements both in soil and plant L. chuanxiong samples were determined by using X-ray fluorescence spectrometry （XRF） and ICP-MS，etc. Spearman correlation test was used to evaluate the correlations of rhizosphere soil and each portion of L. chuanxiong，to screen out “characteristic elements”. Bray-Curtis similarity indexes based on all elements and "characteristic elements" were used for hierarchical cluster analysis of soil samples, to identify the sample range with similar soil element charateristics to the traditional geo-authentic area of L. chuanxiong. Result:K，Mg，Mn and Rb elements were significantly correlated in rhizosphere soil and plant L. chuanxiong. Geographical differentiation of all elements and four characteristic elements showed that there was a highest similarity between geo-authentic area and its neighbor plots S1-S6 (except plot S4) in soil elements distribution, followed by plots S14-S16. The middle section plots S7-S13 in the expansion areas were quite different from the traditional geo-authentic production areas. Conclusion:The soil element characteristics in the study areas were not completely controlled by geographical distance，but demonstrated the "fault" variation in the areas along longitude. This study provides a theoretical basis for the detrrmination of suitable cultivation area for Ligusticum Chuanxiong, and the natural boundary of its geo-authentic area may be extended an additional 60 km southward along longitude beyond the traditional origin. Additionally， K，Mg，Mn and Rb characteristic elements may be the potential markers to evaluate the suitable soil environment for cultivating L. chuanxiong.

Investigation and Evaluation of Cultivation Status of Ligusticum chuanxiong in Sichuan / 中国实验方剂学杂志

Objective::To deeply understand important scientific problems and key technical problems in the cultivation of Chuanxiong, the Chuanxiong cultivation situation of 247 households in 23 towns of 5 counties in Sichuan province were investigated, providing basic data for the standardized cultivation of Chuanxiong. Method::On the basis of reported literatures, a questionnaire survey was conducted on the farmers of cultivating Chuanxiong in main producing areas, field measurements were also preformed, and the cultivation status of Chuanxiong was comprehensively analyzed. Result::The proportion of plain-breeding mainly existed in Pengzhou and Shifang, where per farmer had a small scale of planting areas, was 65%, while the proportion of mountain-breeding mainly existed in Meishan, Qionglai and Dujiangyan, where per proprietor had a large scale of planting areas, was 92%. The planting density and yield of Chuanxiong varied greatly among individuals. The growth period of Chuanxiong in Meishan and Qionglai is about 30 days shorter than that in Pengzhou, Shifang and Dujiangyan. About half of farmers applied base fertilizer and top dressing in spring. The proportion of NPK compound fertilizer input was high (>90%). Chuangxiong has a few diseases and insect pests, the incidence of which in new production areas was obviously lower than that in old production areas. The application of herbicide was not standardized. 52% farmers used Chu cao ling No.1 which was made by agricultural material store owners. The labor cost accounted for the highest proportion (37%) in Chuanxiong cultivation. Conclusion::Now in Sichuan clonal propagated patterns of Chuanxiong were both mountain-breeding and plain-breeding. It is suggested to make clear the differences between mountain-breeding nodes and plain-breeding nodes, and breed high qualities of Chuanxiong nodes. The mechanization research and production of Chuanxiong should be promoted, realizing timely planting and harvesting. Farmers should be guided to increase the application of organic fertilizer, and meanwhile standardize the application of agrochemicals.

Study on the Effects of the Integration of Field Processing and Decoction Piece Processing on Chemical Com- position of Ligusticum chuanxiong Decoction Pieces / 中国药房

OBJECTIVE：To investigate the effects of the integration of field processing and decoction piece processing （hereinafter called “Integration”for short ）on chemical composition of Ligusticum chuanxiong decoction pieces. METHODS ：Fresh L. chuanxiong were collected from Dujiangyan and Pengzhou of Sichuan ；integrated decoction pieces of L. chuanxiong were prepared after washing ，drying in the shade （to about 28％ moisture content ），slicing and drying ；traditional decoction pieces was prepared after drying in the shade ，adding water to moisten （to the core ），slicing and drying. HPLC fingerprints of two kinds of decoction pieces samples （with 10 batches in each type ）were established. The determination was performed on WondaSil C 18 column with mobile phase consisted of 1％ formic acid solution-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was 30 ℃. The detection wavelength was set at 285 nm，and the sample size was 10 μL. Using ligusticolide A as reference ，HPLC fingerprints of 20 batches of samples were drawn. The similarity of the fingerprints was evaluated with Similarity Evaluation System for Chromatographic Fingerprint of TCM （2004A edition ），and then common peaks were confirmed. The contents of chlorogenic acid ，ferulic acid and ligusticolide A were determined by above chromatographic condition. Single factor variance analysis was performed for comparison of the contents. RESULTS ：The similarity of HPLC fingerprints among 20 batches of samples was above 0.900. A total of 16 common peaks were determined ，7 of which were chlorogenic acid ，ferulic acid，ligusticolide Ⅰ，pine cypress ferulinate ，ligusticolide A ，n-butylphthalide and ligustilide ，respectively. The linear range of chlorogenic acid ，ferulic acid and ligusticolide A were 0.008-0.200 mg/mL（r＝0.999 9），0.010-0.140 mg/mL（r＝0.999 2）and 0.100-0.600 mg/mL（r＝0.999 3）；the limits of quantification were 0.002 8，0.000 6 and 0.005 0 mg/mL，respectively；the limits of detection were 0.000 8，0.000 1 and 0.001 0 mg/mL，respectively；RSDs of precision ，reproducibility and stability tests were all lower than 3％，and average recoveries were 96.27％-102.02％（RSD＜2％，n＝6）. The contents of above compositions in the integrated decoction pieces and traditional decoction pieces were（1.677 0±0.311 0），（1.562 7±0.124 5），（9.494 0±1.351 3）mg/g and（1.300 2±0.469 2），（1.388 0±0.209 9），（9.811 7±1.098 9）mg/g，respectively；there was no statistical significance between 2 groups（P＞0.05）. CONCLUSIONS ：The chemical composition of each batch of samples of L. chuanxiong integrated decoction pieces and traditional decoction pieces is consistent ，and the content of index components as chlorogenic acid ，ferulic acid and ligusticolide A in the decoction pieces is not affected by the integration processing. This process is feasible to a certain extent.

Application of quality constant method in evaluation of Ligusticum chuanxiong pieces / 中国中药杂志

The method of classifying the quality grade of traditional Chinese medicine slices with cross section model quality constant was applied to the grade evaluation of Ligusticum chuanxiong pieces,and a reasonable grade standard of L. chuanxiong pieces was established. The purpose is to classify the 15 batches of L. chuanxiong pieces by combining the advantages of traditional grading with modern quality control indicators. By measuring the natural morphological parameters,processing parameters and the intrinsic content of ferulic acid,an important active ingredient,of the 15 batches of L. chuanxiong pieces collected from different manufacturers and different batches of different medicinal materials markets,we can synthesize the results. The mass constants and percentage mass constants are calculated and analyzed based on the above data. The results showed that the quality constants of 15 batches of L. chuanxiong pieces collected ranged from 0.53-3.00; if the percentage mass constants were more than 80%,50%-80% was second-class pieces,and the rest were third-class pieces,the evaluation results were as follows: the quality constants of first-class L. chuanxiong pieces were more than 2.40,the quality constants of second-class L. chuanxiong pieces should be 1.70-2.40,and the quality constants of third-class L. chuanxiong pieces should be less than 1.70. In this paper,the method of dividing the quality constants of the top blade model into different grades is further applied and practiced,which proves that the method is scientific,reasonable and multi-adaptable. At the same time,it enriches the research data of the grade evaluation of L. chuanxiong pieces,provides a useful reference for the promotion of the grade evaluation of L. chuanxiong pieces,and lays an experimental foundation for the next research of the subject group.

High quality and superior effect of Dachuanxiong Formula based on quality constant grading evaluation technology of Chinese medicine pieces / 中国中药杂志

Quality constant is a kind of grading evaluation method of traditional Chinese medicine pieces based on the combination of traditional knowledge and modern quality control. This method has been successfully applied in the grading evaluation of Glycyrrizae Radix et Rhizome pieces and traditional Chinese medicine pieces,and with a positive influence in the industry. With Dachuanxiong Formula for example,different grades of Ligusticum chuanxiong and Gastrodia elata pieces formed high-quality,moderate and qualified Dachuanxiong Formula on the basis of the grading evaluation of the pieces,and the pharmacodynamics method was used to evaluate its efficacy. The results showed that the maximum vascular diastolic rates of Dachuanxiong Formula in the three grades were( 80. 3±5. 2) %,( 67. 0±6. 1) %and( 60. 3±6. 5) %,and the strength of pharmacodynamics was positively correlated with the grade of L. chuanxiong and G. elata pieces.The quality constant technique can objectively and quantitatively classify single decoctions,and has important correlations and prompts for the efficacy of traditional Chinese medicine prescriptions composed of these pieces,with important significance in promoting hierarchical management of the industry,implementing better price for high quality and defining high quality and superior effect.

Distribution and accumulation of inorganic elements in Ligusticum chuanxiong / 中国中药杂志

Aiming at the phenomenon of heavy metal Cd exceeding the standard of Chuanxiong medicinal materials,the accumulation of 12 inorganic elements,including heavy metals,in Ligusticum chuanxiong was studied in this paper. It was found that the contents and distribution of most inorganic elements in the stems and leaves of L. chuanxiong were higher than those in the rhizomes at seedling and shooting stages. The content of most elements in rhizome reached the highest at harvest stage,and the distribution ratio of some elements in rhizome was higher than that in stem and leaf at harvest stage. But rhizome,stem and leaf of L. chuanxiong have relatively stable absorption capacity and enrichment effect on different elements,and are less affected by growth period and position. Rhizomes and stems and leaves of L. chuanxiong were enriched with Cd,and stems and leaves also accumulated Pb at seedling stage and stem stage. The absorption capacity of Pb in stems and leaves of L. chuanxiong was higher than that of rhizomes,and the ability of absorbing Cd was less than that of rhizomes at harvest time. The total uptake of Cd and Pb by L. chuanxiong decreased with the prolongation of growth time,but the proportion of Cd and Pb in rhizome increased,so that the content of Cd and Pb increased with the prolongation of growth time.

The investigation of key factors influencing the extraction process of chuanxiong oil by steam distillation / 中国药学杂志

OBJECTIVE: To investigate the key process parameters that affect the extraction of essential oil from Ligusticum chuanxiong by steam distillation. METHODS: Volatile oil and emulsified aromatic water are the products of the extraction process of volatile oil. The nature of volatile oil will directly affect the formation of emulsified aromatic water. The influence of extraction temperature, the ratio of liquid to medicine, medicinal material granularity process parameters on the stability of the emulsified fragrance water and the yield of volatile oil were analyzed, furtherly the corresponding extraction process control model of volatile oil was established to reveal the key technology and parameter during the process of volatile oil extraction and separation. RESULTS: The relative viscosity, relative density, surface tension and contact angle of Ligusticum chuanxiong oil decreased gradually with the increase of temperature, while the interfacial tension increased with the increase of temperature. The liner model of aromatic water TSI and process parameters was TSI = 0. 877 + 0. 230 × extraction temperature - 0. 024 × comminution particle size + 0. 010 × liquid ratio + 0. 292 × condensation temperature + 0. 776 × collection temperature. The liner model of the physicochemical properties of TSI and volatile oil was TSI = 0. 877 - 0. 170 × density + 0. 098 × viscosity - 0. 301 × surface tension + 0. 695 × interfacial tension - 0. 060 × contact angle. The liner model of volatile oil yield and process parameters was that yield = 5. 065 + 0. 258 × extraction temperature + 0. 127 × particle size + 0. 016 × liquid ratio + 0. 264 × condensation temperature + 0. 264 × collection temperature + 0. 502 × TSI. The liner model with the physicochemical properties of volatile oil was the yield = 5. 065 - 0. 196 × density - 0. 167 × viscosity - 0. 201 × surface tension + 0. 153 × interfacial tension - 0. 065 × contact angle. CONCLUSION: TSI and yield of aromatic water of Ligusti-cum chuanxiong are directly proportional to the extraction temperature and comminution particle size of medicinal materials. The yield of volatile oil shows a significant positive correlation with the TSI value, which can reflect the yield of volatile oil and is a specific index that can evaluate the process parameters objectively.