Study on untargeted metabolomics of Codonopsis pilosula from different producing areas based on ultra-performance liquid chromatography tandem high resolution mass spectrometry / 药学学报

Lu Dangshen, a traditional authentic medicinal material of Codonopsis Radix is mainly produced in Shangdang (Changzhi) area of Shanxi Province. Baitiao Dangshen is mainly produced in Gansu Province. Codonopsis Radix contains many kinds of components such as phenylpropanoids, polyalkynes, alkaloids, terpenes, fatty acids, flavonoids, and so on. At present, the effect of producing areas on its chemical compositions has not been systematically studied. This study analyzed the differences of metabolites among Codonopsis pilosula from different producing areas by UPLC-HRMS. PCA, OPLS-DA coupled with Thermo mzcloud online and local databases were used to compare the overall differences of metabolites among Codonopsis pilosula from different producing areas, and the chemical constituents were identified to further screen and find out the different metabolites and analyze the metabolic pathways by information retrieval in HMDB, PubChem, Chemspider and KEGG databases. The results showed that 72 differential metabolites were identified in this study. There were 15 kinds of up-regulated and 57 kinds of down-regulated metabolites of Lu Dangshen compared with Baitiao Dangshen. The top 30 metabolic pathways were analyzed by KEGG enrichment, and the most important metabolic pathways were phenylpropanoid biosynthesis, which was demonstrated that phenylpropanoid biosynthesis pathway and related intermediate metabolites could be used as the characteristics of distinguishing Lu Dangshen from different habitats of Codonopsis pilosula. The present study provided a basis for analyzing the influence of producing areas on the chemical components of Codonopsis pilosula and reasonably evaluating the quality of Codonopsis Radix, and also provided a new idea for expounding the authenticity of Lu Dangshen.

Quality analysis of Codonopsis pilosula with different commodity specification grades / 中国药房

OBJECTIVE To study the quality of Codonopsis pilosula with different commodity specification grades， and to provide the data support for market transactions， scientific research and clinical use. METHODS According to the classification standard of commodity specification grades of C. pilosula， 17 batches of C. pilosula from different producing areas， origins and commodity specification grades were collected. The contents of tangshenoside Ⅰ， lobetyolin and atractylenolide Ⅲ were determined by HPLC. The contents of alcohol-soluble extracts were determined by hot dipping method stated in general rule 2201 of Chinese Pharmacopeia （part Ⅳ）. The contents of polysaccharide were determined by phenol-sulfuric acid method （calculated by D-glucose anhydrous）. RESULTS For cultivar of C. pilosula， four specifications and three commodity grades of C. pilosula all contained tangshenoside Ⅰ and lobetyolin； Radix C. pilosula from Shanxi of China and C. pilosula from Wenxian County of China， also contained atractylenolide Ⅲ. In terms of the contents of tangshenoside Ⅰ， lobetyolin and atractylenolide Ⅲ， the content of second class was equivalent to that of first class， even better than the first class， while the content of third class was lower than that of first class and second class； the content of tangshenoside Ⅰ was the highest among the two types of wild C. pilosula. The contents of alcohol-soluble extracts and polysaccharides in first class cultivated C. pilosula were higher than those of second class， and the second class was higher than the third class； wild C. pilosula had low content of alcohol-soluble extracts and polysaccharides. CONCLUSIONS The internal quality of C. pilosula is basically consistent with the classification standard of different commodity specification grades； the content of each indicator in first-class and second-class medicinal herb is high， making them high-quality medicinal herbs.

Therapeutic potential of alkaloid extract from Codonopsis Radix in alleviating hepatic lipid accumulation: insights into mitochondrial energy metabolism and endoplasmic reticulum stress regulation in NAFLD mice / 中国天然药物

Alkaloids are a class of naturally occurring bioactive compounds that are widely distributed in various food sources and Traditional Chinese Medicine. This study aimed to investigate the therapeutic effects and underlying mechanisms of alkaloid extract from Codonopsis Radix (ACR) in ameliorating hepatic lipid accumulation in a mouse model of non-alcoholic fatty liver disease (NAFLD) induced by a high-fat diet (HFD). The results revealed that ACR treatment effectively mitigated the abnormal weight gain and hepatic injury associated with HFD. Furthermore, ACR ameliorated the dysregulated lipid metabolism in NAFLD mice, as evidenced by reductions in serum triglyceride, total cholesterol, and low-density lipoprotein levels, accompanied by a concomitant increase in the high-density lipoprotein level. ACR treatment also demonstrated a profound anti-oxidative effect, effectively alleviating HFD-induced oxidative stress and promoting ATP production. These effects were achieved through the up-regulation of the activities of mitochondrial electron transfer chain complexes I, II, IV, and V, in addition to the activation of the AMPK/PGC-1α pathway, suggesting that ACR exhibits therapeutic potential in alleviating the HFD-induced dysregulation of mitochondrial energy metabolism. Moreover, ACR administration mitigated HFD-induced endoplasmic reticulum (ER) stress and suppressed the overexpression of ubiquitin-specific protease 14 (USP14) in NAFLD mice. In summary, the present study provides compelling evidence supporting the hepatoprotective role of ACR in alleviating lipid deposition in NAFLD by improving energy metabolism and reducing oxidative stress and ER stress. These findings warrant further investigation and merit the development of ACR as a potential therapeutic agent for NAFLD.

Effects of stereoscopic traction on photosynthetic characteristics, yield, and quality of Codonopsis pilosula under organic cultivation / 中国中药杂志

To solve the serious problem of stem and leaf shading in the middle and late stage of traditional flat planting of Codonopsis pilosula, this study analyzed the effects of different stereoscopic traction heights on the photosynthetic characteristics and growth of C. pilosula and explored the optimal traction height to improve the yield and quality of C. pilosula. The experiment designed three stereo-scopic traction heights [H1(60 cm), H2(90 cm), and H3(120 cm)] with natural growth without traction as the control(CK). The results showed that the increase in stereoscopic traction heights broadened the growth space of stems and leaves of C. pilosula, enhanced the ventilation effect, significantly increased the average daily net photosynthetic rate of C. pilosula, promoted the absorption of intercellular CO_2, decreased the transpiration rate, and reduced the evaporation of water. Moreover, it effectively avoided the problem of weakened photosynthesis, maintained the carbon balance of individual plants, and promoted the growth and development of the C. pilosula roots. In terms of the seed yield of C. pilosula, it was ranked as H2>H1>H3>CK. To be specific, H1 increased by 213.41% compared with CK, H2 increased by 282.43% compared with CK, and H3 increased by 133.95% compared with CK. The yield and quality of C. pilosula were the highest in the H3 treatment group, with the fresh yield of 6 858.33 kg·hm~(-2), 50.59% higher than CK, dry yield of 2 398.33 kg·hm~(-2), 76.54% higher than CK, and lobetyolin content of 0.56 mg·g~(-1), 45.22% higher than CK. Therefore, the stereoscopic traction height has a great influence on the photosynthetic characteristics, yield, and quality of C. pilosula. Particularly, the yield and quality of C. pilosula can be optimized and improved in the traction height treatment of H3(120 cm). This planting method is worth popularizing and applying in the cultivated management of C. pilosula.

Research progress of Codonopsis Radix and prediction of its Q-markers / 中国中药杂志

Codonopsis Radix is a traditional tonic medicine commonly used in China, which has the effects of strengthening the spleen and tonifying the lung, as well as nourishing blood and engendering liquid. The chemical constituents of Codonopsis species are mainly polyacetylenes, alkaloids, phenylpropanoids, lignans, terpenoids and saponins, flavonoids, steroids, organic acids, saccharides, and so on. Modern pharmacological studies showed that Codonopsis Radix also has a variety of pharmacological effects such as enhancing body immunity, protecting gastrointestinal mucosa and resisting ulcers, promoting hematopoietic function, regulating blood sugar, and delaying aging. In this paper, the chemical constituents of Codonopsis species and the pharmacological effects of Codonopsis Radix were summarized, and on this basis, the quality markers of Codonopsis Radix were analyzed. It was predicted that lobetyolin, tangshenoside I, codonopyrrolidium A, and the oligosaccharides were the possible Q-markers of Codonopsis Radix. This paper will provide scientific references for the quality evaluation and profound research and the development of Codonopsis Radix.

Chemical Components Distribution and Transcriptome Analysis of Different Tissues from Codonopsis pilosula / 中国实验方剂学杂志

ObjectiveThe transcriptome characteristics of different tissues of Codonopsis pilosula were analyzed to illustrate the genetic basis of the accumulation of active ingredients in the root of C. pilosula， and to provide theoretical basis for its high-quality production and cultivation. MethodDifferent tissues of C. pilosula at flowering stage were selected as experimental materials， and the contents of tangshenoside Ⅰ， lobetyolin and atractylenolide Ⅲ were detected by high performance liquid chromatography（HPLC）. RNA-Seq was used to perform transcriptome sequencing of different tissues， and the differentially expressed genes were screened and analyzed by Gene Ontology（GO） and Kyoto Gene and Encyclopedia of Genes and Genomes（KEGG） enrichment analysis， in order to explore the characteristics of active compound distribution and the transcriptional profiles. ResultThe contents of polysaccharides and tangshenoside Ⅰ in the root of C. pilosula were significantly higher than those in other tissues. The transcriptional profiles of the root were significantly different from those of stem， leaf and flower. Cluster analysis， GO and KEGG enrichment analysis of differential gene expression showed that the differential expression genes were mainly enriched in flavonoid and phenylpropanoid biosynthesis， sucrose-starch metabolism， plant hormone signal transduction， plant-pathogen interaction， mitogen-activated protein kinase（MAPK） cascade signal transduction， Adenosine triphosphate（ATP）-binding cassette（ABC） transporter and other pathways. The expression of genes related to biosynthesis of phenylpropanoid compounds were significantly up-regulated in the roots and flowers， and ABC transporter proteins were mostly highly expressed in the flowers. The expression of key enzyme genes for polysaccharide synthesis， such as sucrose：sucrose 1-fructosyltransferase（1-SST） and fructan 1-exohydrolase（1-Feh）， were significantly up-regulated in the roots， and a large number of stress-responsive genes closely related to the accumulation of secondary metabolites were significantly up-regulated in the roots. ConclusionThe active compound content and transcriptional profiles in C. pilosula roots were significantly different from those in stem， leaf， flower and other tissues， showing tissue specificity. Meanwhile， the genes related to stress response and biosynthesis of active compound， such as fructan and phenylpropanoid compounds， were up-regulated in roots of C. pilosula.

Second metabolites comparative analysis of Codonopsis pilosula (Franch.) Nannf. from different origins based on extensively targeted metabolomics / 药学学报

We performed an extensively targeting metabolomic detecting using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) to compare the secondary metabolites in Dang shen [Codonopsis pilosula (Franch.) Nannf.] from Shanxi and Gansu provinces. The findings showed that 161 secondary metabolites in 6 groups (phenolic acids, flavonoids, lignans and coumarins, alkaloids, terpenoids, others) were found from Dang shen in Changzhi city of Shanxi province and Dingxi city of Gansu province. There were 98 secondary metabolites which is differed significantly. In comparison to Dingxi city, 33 different secondary metabolites of Dang shen in Changzhi city had a greater relative content, whereas relative content of 65 different metabolites in Dingxi city was higher. Metabolic pathway enrichment analysis revealed that phenolic acids and flavonoids were significantly different in the secondary metabolites of Dang shen from different producing places. This may be one of the reasons for the difference in the quality of Dang shen in Shanxi and Gansu provinces. This work compared and analyzed the secondary metabolites of Dang shen from Dingxi city in Gansu province and Changzhi city in Shanxi province for the first time, which lays the foundation for further study on the quality of Dang shen.

Identification and analysis of glycosyltransferase in Codonopsis pilosula from different regions based on transcriptome / 药学学报

Lu Dangshen is the geoherb in Shanxi Province. The content of Codonopsis pilosula polysaccharides (CPP) in Lu Dangshen is more than that in other Codonopsis Radix from other regions. Glycosyltransferase is the key enzyme for the synthesis of bioactive components, such as CPP and tangshenoside I. Based on the transcriptome data of C. pilosula [Codonopsis pilosula (Franch.) Nannf.] from different producing areas, this study carried out functional annotation of GO and KEGG, conservative domain analysis, phylogenetic tree analysis and expression pattern analysis of glycosyltransferase genes in C. pilosula to provides a theoretical basis for exploring the mechanism of genuineness formation in Lu Dangshen. In this study, 98 glycosyltransferase genes were screened and identified, which belonged to GT family 1, GT family 2, GT family 90 and other families. By GO functional annotation, it was found that most of the glycosyltransferase genes had catalytic activity. Analysis of KEGG functional annotation showed that C. pilosula glycosyltransferase was mainly involved in glycan organism and terpenoid and polyketone metabolism. Among them, conserved domain of 42 glycosyltransferase genes in GT family 1 was [X]-W-[2X]-Q-[3X]-[LH]-[5X]-[FLTHCGWNS]-[2X]-E-[4X]-[GVP]-[4X]-P-[4X]-Q-[2X]-[NAK]. Phylogenetic tree analysis based on the glycosyltransferase sequence in Arabidopsis thaliana showed that C. pilosula glycosyltransferases were mainly located in Arabidopsis thaliana UGT73, 72 and 85 branches. Gene expression pattern analysis showed that expression of CpUGT73AH2 was higher in Lu Dangshen than that in Baitiaodang and could respond to drought and low temperature stress. In conclusion, a glycosyltransferase gene CpUGT73AH2, which is involved in the metabolism of terpenoids and polyketides and can respond to environmental stress, was screened from the C. pilosula glycosyltransferase family 1, which was used to further study the role of C. pilosula glycosyltransferase in Lu Dangshen. It laid a theoretical foundation for further study on the role of C. pilosula glycosyltransferase in the formation of Lu Dangshen.

Molecular Identification of Tibetan Codonopsis Plants Based on ITS2 and psbA-trnH Barcode / 中国实验方剂学杂志

ObjectiveTo identify the molecular biology of various species of Tibetan Codonopsis plants based on internal transcribed spacer（ITS）2 and psbA-trnH sequence barcode technology. MethodThe genomic DNA of 28 Tibetan Codonopsis plant samples from four species （Codonopsis canescens，C. foetens subsp. nervosa，C. pilosula， and C. thalictrifolia var. mollis） were extracted，and the ITS2 and psbA-trnH sequences were amplified and sequenced. The related sequences of 81 Tibetan Codonopsis plant samples belonging to 15 species were downloaded from GenBank， and MEGA 6.0 was used for sequence comparison and mutation site analysis. The GC content and genetic distance within and between species were calculated. Additionally， phylogenetic trees were constructed by maximum likelihood （ML） method， neighbor-joining （NJ） method，and unweighted pair-group method with arithmetic means （UPGMA） . ResultAccording to the mutation site，C. canescens， C. pilosula，C. pilosula subsp. tangshen， C. pilosula var. modesta，C. bhutanica，C. clematidea，C. lanceolata，C. subglobosa and C. foetens were distinguished. In the phylogenetic trees，the optimal clustering effects for ITS2 and psbA-trnH sequences were obtained using the ML method and the UPGMA method， respectively， and 12 species were effectively clustered. ConclusionITS2 and psbA-trnH sequences have a high identification rate for species of single origin，but there are still some limitations in identifying variants and original variants. This study provides basis for the identification of affinity relationship and clinical safety of Tibetan Codonopsis plants.

Correlation Between Functional Components in Codonopsis pilosula Roots and Soil Factors / 中国实验方剂学杂志

ObjectiveTo explore the correlation between the content of 4 functional components in Codonopsis pilosula roots from different areas and soil factors， and thereby to lay a theoretical basis for soil ecological regulation and improvement of quality of C. pilosula roots. MethodThe content of lobetyolin， atractylenolide Ⅲ， alcohol extract， and polysaccharides， as well as soil fertility and 16 soil factors in 24 batches of samples from different producing areas were determined. Principal component analysis （PCA） and Pearson's correlation analysis were used to explore the key soil factors leading to the variation of chemical component content in C. pilosula roots. ResultThe content of lobetyolin and atractylenolide Ⅲ in samples from Longxi was the highest， and the content of polysaccharides peaked in samples from Huguan. The content of lobetyolin was in positive correlation with soil total nitrogen， total phosphorus， total potassium， alkali-hydrolyzable nitrogen， and available potassium （P<0.01）， as well as soil organic matter， pH， available manganese， and available zinc （P<0.05）. There was a positive correlation between pH and atractylenolide Ⅲ content （P<0.05）. Soil total potassium was in positive correlation with alcohol extract and polysaccharide content （P<0.01）. Soil available zinc was positively correlated with alcohol extract and the polysaccharide content （P<0.05）. Sample sites with higher PCA scores were Pingshun， Huguan， and Longxi， which were significantly positively correlated with the content of polysaccharides in C. pilosula roots in different habitats. ConclusionThe content of functional components in C. pilosula roots can be improved by raising soil organic matter content and applying specific fertilizers.

Herbal Textual Research on Local Varieties of Codonopsis Radix / 中国实验方剂学杂志

Codonopsis Radix， one of the bulk commodities， has been commonly used for tonification in clinical practice. Apart from the medicinal purpose， it can also be utilized as food. Among the multiple local varieties， the ones called "Luduiduoji" in Tibetan medicine cannot be neglected， which have frequently been adopted for diminishing inflammation and swelling， invigorating spleen and stomach， and tonifying Qi， etc. Considering its complex origins and frequent substitution by or confusion with other medicinal materials， this paper reviewed the Si Bu Yi Dian， Jingzhu Bencao， ministerial and local standards， modern literature on Tibetan medicine， and the results of field investigation in major Tibetan medicine hospitals and medicinal material markets of Sichuan， Qinghai and Tibet to figure out the name， original plants， medicinal parts， effects， and local varieties of Codonopsis Radix in Tibetan medicine. The results showed that the names of local varieties were diverse， many of which were transliterated into Tibetan， with "Luduiduoji" being most well-known. It was derived from 14 species in genus Codonopsis and one in Adenophora of family Campanulaceae， with Codonopsis foetens subsp. nervosa， C. thalictrifolia var. mollis， C. canescens， C. alpina， and C. pilosula being the main species. According to literature records， the roots， aerial parts， and whole plants could all be employed as medicine， but there were certain differences in their clinical applications. At present， in order to protect the medicinal resources， Tibetan medical workers mostly collect the aerial parts， which are applicable to patients with yellow water， rheumatism， Gamba disease， and leprosy. This literature review of local varieties for Codonopsis Radix and textual research on their original plants are of great significance for elevating the standard， accelerating the pharmacodynamic research， expanding the sources and promoting the rational use of Codonopsis Radix.

Influence of Yuanzhisan Containing Ginseng Radix et Rhizoma or Codonopsis Radix on Memory Disorder / 中国实验方剂学杂志

Objective:To explore the effects of Yuanzhisan （YZS） containing Ginseng Radix et Rhizoma （YZSR） or Codonopsis Radix （YZSD） on memory disorder based on network and experimental pharmacology. Method:The active components and targets of YZS were retrieved from the component database and literature， and the targets of memory disorder from the disease databases. The intersection targets revealed by Veen diagram were subjected to pathway analysis. The common active components of YZSR and YZSD were molecularly docked onto the core targets. Scopolamine hydrobromide was used to establish the memory disorder model， which was employed in the behavioral experiments for evaluating the effect of YZSR and YZSD on memory disorder. Result:There existed 33 active components for Ginseng Radix et Rhizoma and 31 for Codonopsis Radix， with four common active components and 380 common targets. YZSR contained 85 active components and 790 drug targets， and YZSD 81 active components and 781 drug targets. The mapping of 425 memory disorder targets with those of YZSD and YZSD yielded 133 and 130 intersection targets， respectively. The metabolic pathways involved calcium ion signaling pathway， hypoxia-inducible factor-1 （HIF-1） signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， etc. As revealed by molecular docking， the binding energy of common active components to the targets was negative， and the binding effect of frutinone A was the best. Behavioral experiment results showed that both YZSR and YZSD alleviated the memory disorder. In the step-down test， the number of errors in the YZSD group was significant lower than that in the model group （<italic>P</italic><0.01）. In Morris water maze test， the movement distance of the YZSD group was remarkably shortened in comparison with that of the model group （<italic>P</italic><0.05）. In the open field test， the movement distances of both the YZSR and YZSD group were shortened in contrast to that in the normal group （<italic>P</italic><0.05）. Conclusion:YZS had a certain effect on memory disorder. There are similarities and differences between YZSR and YZSD in the treatment of memory disorder.

Enhancement of Quality and Yield in Codonopsis pilosula Seedlings by Soil Fumigation with Dazomet / 中国实验方剂学杂志

Objective:To explore the high-efficiency and high-quality seedling raising method of <italic>Codonopsis pilosula</italic>. Method:In the main production area of <italic>C. pilosula</italic> in the Tanchang county，Gansu province，after the soil was fumigated with dazomet （containing 98% methylisothiocyanate）， four varieties of <italic>C. pilosula</italic> seedlings were raised. The dynamic change in growth and differences in quality and yield of <italic>C. pilosula</italic> seedlings after emergence were regularly determined. Result:① The soil enzyme activity was first inhibited and then restored by soil fumigation，which increased the root length of <italic>C. pilosula</italic> seedlings by 9.8%. Besides， the field growth indexes such as plant height，plant width，stem length，stem diameter，number of branches，number of nodes，number of leaves， and fitted leaf area increased in varying degrees，and the plant height showed an "S"-shaped growth trend. "Gandang No.1" and "Gandang No.2" grew better than "Weidang No.1" and "Tanchang control". ② Fumigation reduced the incidence rate of <italic>C. pilosula</italic> root in the field by 4.9%，and the incidence rates of "Gandang No.1" and "Gandang No.2" were significantly lower than those of "Weidang No.1" and "Tanchang control". ③ Fumigation increased the total number of <italic>C. pilosula </italic>seedlings by 6.15×10⁵ plants·hm^-2，of which the number of primary seedlings increased by 45.3% and that of secondary seedlings increased by 42.2%. ④ Fumigation increased the seedling yield of <italic>C. pilosula</italic> by 42.4%. It showed the most significant effect on the yield of "Gandang No.2"，which increased by 61.8%， and the weakest effect on the yield of "Gandang No.1"，which increased by 15.4%. ⑤ Comprehensive analysis showed that the quality and yield of <italic>C. pilosula</italic> seedlings in the fumigation area were better than those in the non-fumigation area. Conclusion:The results showed that soil fumigation showed a promoting effect on the seedling yield of <italic>C. pilosula</italic> in spite of different effects achieved in terms of different varieties.

Development and Application of Mitochondrial and Chloroplast Microsatellite Markers for Codonopsis Plants / 中国实验方剂学杂志

Objective:To develop the specific molecular markers of <italic>Codonopsis</italic> plants and better identify their germplasm resources considering the significant difference in active ingredients of Codonopsis Radix from various origins and producing areas. Method:Such bioinformatics software as Primer 5.0， NTSYS-pc 2.10e， and PopGene 32 were used for searching the simple sequence repeat （SSR） markers of <italic>C. minima </italic>chloroplast genome， <italic>C. tsinlingensis</italic> chloroplast， and <italic>C. lanceolata </italic>mitochondrial sequences， and 120 pairs of SSR primers were designed by Primer 5.0. Then 16 pairs of cpSSR primers and 10 pairs of mtSSR primers with good screening effect and high polymorphism were selected for analyzing the interspecific versatility of 20 samples. Result:The results showed that 66 cpSSR primer sites and 26 mtSSR sites were identified from the genome sequences， with 86.20% of single nucleotide， 6.9% of dinucleotide， and 6.9% of trinucleotide for <italic>C. minima </italic>chloroplast， 83.78% of single nucleotide，13.51% of dinucleotide， and 2.71% of trinucleotide for <italic>C. tsinlingensis</italic> chloroplast， and 46.15% of single nucleotide and 53.85% of dinucleotide for <italic>C. lanceolata </italic>mitochondria. As demonstrated by polymerase chain reaction （PCR） identification results， the developed 26 pairs of SSR primers had good applicability in the genus<italic> Codonopsis</italic>. The analysis by NTSYS-pc 2.10e revealed that the genetic similarity coefficients of 20 samples were within the range of 0.38-1.00， and they were divided into two subgroups at a threshold of 0.69. Four pairs of polymorphic primers were screened out in the diversity analysis of 20 samples using PopGene 32. The number of observed alleles （<italic>Na</italic>） was 12， and the effective number of alleles （<italic>Ne</italic>） ranged from 1.362 9 to 2.605 9. The percentage of polymorphic loci （PPL） at each site was 100%， and the average values of genetic parameters<italic> Ho</italic>， <italic>He</italic>， and <italic>I</italic> at each site were 0.555 8， 0.444 2， and 0.753 2， respectively， indicating high polymorphism at each site. The screened four pairs of primers were utilized for DNA fingerprinting of the 20 samples， and it was found that the DNA fingerprints enabled the identification of these 20 samples. Conclusion:This study has provided a molecular basis for the study of the genetic relationship between plants in species <italic>Codonopsis</italic> and the intraspecific genetic differentiation.

Metabolomics Analysis of Codonopsis pilosula Under Drought Stress Based on LC-MS / 中国实验方剂学杂志

Objective:The biological mechanism of <italic>Codonopsis pilosula</italic> adaptation to drought was explored by determining the root metabolome of <italic>C. pilosula</italic> during harvesting. Method:Non-targeted metabonomics LC-MS was used to screen differential metabolites by multivariate statistical analysis，univariate statistical analysis and metabolic pathway enrichment analysis. Result:①There were 274 metabolites in LD vs CK group，142 of which were up-regulated and 132 of which were down-regulated. There were 284 metabolites with significant difference in MD vs CK group，of which 157 were up-regulated and 127 were down-regulated. There were 317 metabolites with significant difference in SD vs CK group，of which 133 were up-regulated and 184 were down-regulated. ②Differential metabolites were annotated into kyoto Encyclopedia of Gene and Genomes （KEGG） database and 82 differential metabolic pathways were obtained，among which sphingolipids metabolism was significantly enriched （<italic>P</italic><0.01）.Metabolism of arginine and proline，tryptophan，alanine，galactose，nicotinic acid and nicotinamide，cysteine and methionine，arachidonic acid，linolenic acid and glycerides were significantly enriched in different metabolite pathways （<italic>P</italic><0.05）. ③The metabolites of the three comparison groups before and after enrichment were classified and analyzed. It was found that they were mainly concentrated in fatty acyls group，carboxylic acid and derivatives，and organ oxygen compounds，followed by sphingolipids，indoles and derivatives，organonitrogen compounds，glycerophospholipids，pyridines and derivatives，peptidomimetics，glycerolipids and so on.In the drought stress of <italic>C. codonopsis</italic>，carbohydrate related metabolites were mainly up-regulated，lipid related metabolites were mainly down-regulated，and all other metabolites were up-regulated. Conclusion:The changes of metabolites in the roots of <italic>C. pilosula</italic> under drought stress were elucidated. carbohydrate and lipid-related metabolites were the main products of <italic>C. pilosula</italic> under drought stress，and these metabolites may be the main reason to improve the ability of <italic>C. pilosula</italic> to resist drought，which laid a foundation for further study on the mechanism of <italic>C. pilosula</italic> to resist drought.

Preliminary Investigation of Resources and Medicinal Use of Codonopsis canescens / 中国实验方剂学杂志

Objective:To study the resources and medicinal use of Tibetan medicine Codonopsis canescens，analyze its suitable habitat，and protect its wild resources，in order to provide an important reference basis for further rational development and utilization. Method:Combined investigation methods of literature textual research，visiting investigation and on-the-spot investigation were adopted. Result:In this paper，we investigated the resources and medicinal use of C. canescens in 27 cities and counties of Sichuan，Qinghai and Tibet. Compared with historical records of C. canescens，the results showed that there was no obvious change in the distribution range，and the distribution was uneven in different places. Ganzi Prefecture， Sichuan， boasts rich resources； in particular， Luhuo county has as high as 90-120 plants/100 m2， it is estimated that the reserves per unit area of root C. canescens in this area were 3 445.32 g·m-2，which totaled 1 279.18 ton（1 ton=1 000 kg）. In history，the root is the main medicinal part. However， in recent years，in order to protect resources，the aboveground part is frequently used. The clinical efficacy of Tibetan medicine is mainly to tonify Qi and activate blood circulation. It can be combined with other drugs to treat rheumatism，sores，furuncle，carbuncle and other diseases. However，the plants of Codonopsis are mixed with C. canescens in each Tibetan area，and the mixed varieties are determined according to the growing varieties of the same genus in this area. Conclusion:The reserves of wild resources of C. canescens are decreasing with the increase of clinical use and market demand of Tibetan medicine. There is serious mixture of Codonopsis plants in Tibetan areas. It is suggested paying attention to resource protection，carrying out artificial planting，establishing the standard for the medicinal use of Tibetan medicine，and promote rational and sustainable utilization of resources.

Application of Relative Quality Constant in Grade Evaluation of Codonopsis Radix Slices / 中国实验方剂学杂志

Objective:To establish the grade evaluation method for Codonopsis Radix slices based relative quality constants， in order to provide scientific theoretical basis for grading of Codonopsis Radix slices. Method:Through literature and market research，the main production areas of Codonopsis Radix slices were determined，and 67 batches of Ludangshen slices（52 batches） and Baitiaodang slices （15 batches） were collected. The appearance traits （average quality and average thickness of Codonopsis Radix slices） were observed and measured. According to Chinese Pharmacopoeia （2015 edition）， the extract and the content of Codonopsis pilosula polysaccharide was determined by phenol-sulphoacid method. Then the relative quality constant was calculated，and the results of grade evaluation were evaluated through systematic cluster analysis and correlation analysis. Result:Relative quality constants of 67 batches of Codonopsis Radix slices were between 0.32-2.97. If these samples were divided into three grades：the first-grade relative quality constants were greater than or equal to 2.08，the second grade was greater than or equal to 0.89 but less than 2.08，while the third grade was less than 0.89. The results of systematic cluster analysis showed that 67 batches of Codonopsis Radix slices were clustered into 3 categories，and the results were basically consistent with the classification. The correlation analysis showed a positive correlation between the content of the extracts and the polysaccharide content （P<0.05）. Conclusion:This method links the extrinsic characteristics to the intrinsic quality，and objectively grade Codonopsis Radix slices， so as to provide a basis for its grade standards.

Study on the Tonifying Spleen Effect of Codonopsis pilosula Polysaccharides on Sennae Folium -induced Spleen- deficiency Model Mice / 中国药房

OBJECTIVE： To study the tonifying spl een effect of Codonopsis pilosula polysaccharides （CPP） on spleen-deficiency model mice. METHODS ：Totally 60 ICR male mice were randomized into blank group ，model group ，CPP high-dose，medium-dose and low-dose groups [ 1.6，0.8，0.4 g/（kg·d）]，Sijunzi tang （SJZT）group [ 30 g/（kg·d），by crude drug] ， with 10 mice in each group. Except for blank group ，other groups were given Sennae Folium solution intragastrically （0.4 g/d）to establish spleen-deficiency model. After modeling ，administration groups were given relevant drug intragastrically ，and blank group and model group were given 20 mL/kg normal saline intragastrically ，once a day ，for consecutive 6 weeks. After last administration，body weight of mice in each group was measured and their general behavioral characteristics were observed. The red blood cell count in whole blood were recorded ，and the contents of D-xylose，gastrin（GAS），motilin（MTL），somatostatin （SS），vasoactive intestinal peptide （VIP），amylase（AMS），immunoglobulin（IgG and IgM ）and lipopolysaccharide （LPS）in serum were determined ；mRNA and protein expressions of Claudin and Occludin in colon tissues of mice were also detected. RESULTS：Compared with blank group ，body weight of mice in model group was significantly decreased （P＜0.05），and the spleen deficiency symptoms such as sparse back hair and no luster appeared ；the red blood cell count in whole blood ，serum contents of D-xylose，SS，VIP，AMS，IgG and IgM ，mRNA and protein expressions of Claudin and Occludin were significantly decreased（P＜0.05 or P＜0.01）；serum contents of GAS ，MTL and LPS were significantly increased （P＜0.01）. Compared with model group ，the body weight of mice were increased significantly in CPP high-dose group （P＜0.05），and spleen-deficiency symptom was improved significantly. Except for the serum contents of D-xylose and IgM ，the protein expressions of Claudin and Occludin in colon tissue had no statistical significance in CPP low-dose group （P＞0.05），above indexes of other groups w ere improved significantly （P＜0.05 or P＜0.01）. CONCLUSION ：CPP can i mprove spleen-defic iency model mice induced by Sennae Folium，the mechanism of which may be associated with regulating gastrointestinal hormone secretion ， enhancing immunity and protecting intestinal mucosal barrier.

Effects of Codonopsis tralictrifolia Extract on Collagen-induced Arthritis Model Rats and Its Mechanism Study / 中国药房

OBJECTIVE：To study the effec ts of Tibetan Codonopsis tralictrifolia extract （called“ZDS”for short ） on collagen-induced arthritis （CIA）model rats and its mechanism. METHODS ：Eight of 48 rats were randomly selected as normal control group （normal saline ），and the remaining 40 rats were used to establish CIA model. After successful modeling ，the rats were randomly divided into model group （normal saline ），ZDS low-dose ，medium-dose and high-dose groups （0.44，0.88，1.76 g/kg，by crude drug ），dexamethasone group （positive control ，0.002 5 g/kg），with 8 rats in each group. They were given relevant medicine intragastrically ，the volume of 400 μL，once a day ，for consecutive 28 days. The body weight of rats were weighed before medication （0 d），7，14，21 and 28 days after medication ；and arthritis indexes were scored. The pathological changes of the knee joint synoviual tissue were observed after last medication. The thymus index ，spleen index ，the levels of serum inflammatory factors （IL-1β，TNF-α，IL-6），protein expressions of NF-κB p65，p-NF-κB p65，IκB and p-IκB in synovial tissue were detected. RESULTS ：Compared with normal control group ，the body weight （14，21，28 days after administration ）of rats in model group was significantly reduced （P＜0.05）；the arthritis index score （before administration and different administration time）was significantly increased （P＜0.05）；the joint synovial tissue was pathologically damaged ；the thymus and spleen index ， inflammation factor level ，the protein expression of p-NF-κB p65 and p-IκB were increased significantly（P＜0.05），while the protein expression of IκB was decreased significantly（P＜0.05）. Compared with model group ，the level of IL- 1β was decreased significantly in ZDS low-dose group （P＜0.05）. Body weight of rats （21，28 days after administration ）were increased significantly in ZDS medium-dose and high-dose groups ，dexamethasone group （P＜0.05），while arthritis index score （14，21，28 days after administration ） was decreased significantly （P＜0.05）. The pathological damage of joint synovial tissue was significantly relieved ；thymus and spleen index ，inflammation factor level ，the protein expression of p-NF-κB p65 and p-IκB were significantly reduced，while the protein expression of IκB was significantly increased （P＜0.05）. CONCLUSIONS ：ZDS can improve CIA model rats to some extent ，and its mechanism may be related to the inhibition of NF-κB signaling pathway.

Network pharmacology unveils spleen-fortifying effect of Codonopsis Radix on different gastric diseases based on theory of “same treatment for different diseases” in traditional Chinese medicine / 中草药·英文版

Objective: “Same treatment for different diseases” is a unique treatment strategy under the guidance of traditional Chinese medicine (TCM) theory. Codonopsis Radix (Codonopsis pilosula, Dangshen in Chinese) with spleen-fortifying effect was employed to understand the strategy of “Same treatment for different diseases”, based on its common mechanism in the treatment of gastric diseases including gastric ulcer, gastritis and gastric cancer via network pharmacology research. Methods: Network pharmacology research methods were used to analyze the interaction network and potential mechanisms of Dangshen in treating gastric ulcer, gastritis and gastric cancer. The active components and their target proteins of Dangshen were integrated from TCMSP, BATMAN-TCM databases. The targets of gastric ulcer, gastritis and gastric cancer were collected through GeneCards, PubMed, TDD and DisGeNET Database. Through screening, the key components and the key targets of Dangshen in treating gastric ulcer, gastritis and gastric cancer were obtained. After KEGG pathway analysis and GO analysis, the important pathways and biological processes were analyzed. Results: Through data and literature mining, the common and specific pharmaceutical effects and mechanism of Dangshen were summarized in these three gastric lesions. It was shown that Dangshen mainly acted on gastric ulcer, gastritis and gastric cancer through the overall regulation of the PI3K-AKT signaling pathway. With the development of the disease, it will gradually increase the control of inflammation through TNF, NF-κB and other inflammation-related signaling pathways to reduce inflammatory damage. For tumorigenesis, it pays more attention to inhibiting the ErbB signaling pathways to reduce the proliferation and migration of tumor cells. In addition, Dangshen's regulation of HIF-1 signaling pathway may also be beneficial for the treatment of gastric ulcer, gastritis and gastric cancer. Conclusion: Dangshen achieves spleen-fortifying effect on gastric diseases including gastric ulcer, gastritis and gastric cancer through multiple targets in multiple pathways, especially PI3K-AKT pathway and HIF-1 pathway. It could provide a scientific basis for understanding the strategy of “Same treatment for different diseases” in traditional Chinese medicine.