Total Saponins in Paridis Rhizoma： A Review / 中国实验方剂学杂志

Paridis Rhizoma possesses the functions of clearing heat and detoxifying， alleviating swelling and relieving pain， cooling the liver and calming the convulsion. Saponins are the main active components of Paridis Rhizoma. Studies have shown that total saponins in Paridis Rhizoma have obvious inhibitory effect on solid tumors such as breast cancer， lung cancer， gastric cancer， and liver cancer and non-solid tumors such as leukemia. The saponins may exert the anti-tumor effects by inhibiting the proliferation， migration， and invasion of tumor cells， regulating cell cycle， inducing apoptotic and non-apoptotic death pathways， and regulating metabolism and tumor microenvironment. Furthermore， total saponins in Paridis Rhizoma showed anti-inflammatory， antioxidant， antimicrobial， hemostatic， and uterus-contracting activities. At the same time， they may induce apoptosis of normal cells， inflammation and oxidative stress， and metabolic disorders. In recent years， the reports of liver injury， reproductive injury， gastrointestinal injury， hemolysis， and other adverse reactions caused by total saponins in Paridis Rhizoma have been increasing. Pharmacokinetic studies have shown that there are significant differences in the metabolism of total saponins in Paridis Rhizoma administrated in different ways. Injection has a fast clearance rate， while oral administration may have hepatoenteric circulation. Meanwhile， due to the low solubility and activation of P-glycoprotein （P-gp） molecular pump， the prototype absorption， intestinal permeability， and recovery rate of total saponins in Paridis Rhizoma are poor， which affects the bioavailability. The bioavailability can be improved to some extent by preparing new dosage forms or new drug delivery systems with advanced technology. This paper reviews the pharmacological effect， pharmacokinetics， and adverse reactions of Rhizoma Paridis total saponins by searching the China National Knowledge Infrastructure （CNKI）， VIP， and Web of Science with ''Rhizoma Paridis total saponins'' as the keywords， hoping to provide references for the research， development， and clinical application of such components.

New steroidal saponins from aerial parts of Paris polyphylla var. chinensis / 中国中药杂志

The shortage of Paridis Rhizoma promotes comprehensive utilization and development research of waste aerial parts of the original plant. The chemical compositions of the aerial parts of Paris polyphylla var. chinensis were clarified based on the ultrahigh performance liquid chromatography tandem quadrupoles time of flight mass spectrometry(UPLC-QTOF-MS/MS) in the previous investigation, and a series of flavonoids and steroidal saponins were isolated. The present study continued the isolation and structure identification of the new potential compounds discovered based on UPLC-QTOF-MS/MS. By using silica gel, ODS, flash rapid preparation, and other column chromatography techniques, combined with prepared high performance liquid chromatography, five compounds were isolated from the 75% ethanol extract of the aerial parts of P. polyphylla var. chinensis, and their structures were identified by spectral data combined with chemical transformations, respectively, as(23S,25R)-23,27-dihydroxy-diosgenin-3-O-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→3)]-β-D-glucopyranoside(1),(25R)-26-O-β-D-glucopyranosyl-furost-5-en-3β,22α,26-triol-3-O-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranoside(2),(25R)-27-O-β-D-glucopyranosyl-5-en-3β,27-dihydroxyspirost-3-O-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranoside(3),(25R)-27-O-β-D-glucopyranosyl-5-en-3β,27-dihydroxyspirost-3-O-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→3)]-β-D-glucopyranoside(4), and aculeatiside A(5). Among them, compounds 1-4 were new ones, and compound 5 was isolated from P. polyphylla var. chinensis for the first time.

Anti-colorectal cancer mechanism of Astragali Radix-Curcumae Rhizoma-Paridis Rhizoma based on network pharmacology and experimental verification / 中国中药杂志

The present study explored the underlying mechanism of Astragali Radix-Curcumae Rhizoma-Paridis Rhizoma(AR-CR-PR) in the treatment of colorectal cancer(CRC) by network pharmacology and molecular docking and animal tests and verified the core targets based on the orthotopic transplantation model in nude mice. The active components of AR-CR-PR were retrieved from databases such as TCMSP. The targets of drugs and the disease were obtained from PubChem, SwissTargetPrediction, TTD, and DrugBank, and the intersection targets were imported into STRING for the analysis of the protein-protein interaction(PPI). Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) analyses were performed through DAVID. AutoDock Vina was used to perform molecular docking and binding ability prediction between the active components and the core targets. The effects of AR-CR-PR on tumor growth, metastasis, and phosphorylation of core target proteins in tumor tissues based on the orthotopic transplantation model in nude mice. As revealed by network pharmacology, AR-CR-PR contained nine core components, such as quercetin, curcumin, and β-ecdysone, and the key targets included protein kinase B(AKT1), mitogen-activated protein kinase 3(MAPK3), MAPK1, and epithelial growth factor receptor(EGFR), which was indicated that the anti-CRC effect of AR-CR-PR was presumedly achieved by regulating tumor cell proliferation, apoptosis, migration, and angiogenesis through PI3 K-AKT, MAPK and other signaling pathways. The results of molecular docking showed that the nine core components had strong binding abilities to AKT1 and MAPK3. The results in vivo showed that AR-CR-PR could reduce the volume of the orthotopic tumor, inhibit liver metastasis, and decrease the phosphorylation of AKT1 and MAPK3 in the CRC model. The mechanism of AR-CR-PR in the intervention of CRC may be related to the activation of PI3 K-AKT and MAPK signaling pathway. This study provides a scientific basis for the clinical application of AR-CR-PR in the treatment of CRC and ideas for modern research on AR-CR-PR.

Separation and structural elucidation of C25 epimers of furostanol saponin from aerial parts of Paris polyphylla var. chinensis / 中国中药杂志

Paris polyphylla var. chinensis(PPC) is used as one of the origin plants of Paridis Rhizoma described in the Chinese Pharmacopoeia(2020 edition). Its resources shortage makes the planting scale gradually expand, and plenty of aerial parts are abandoned because of not being effectively used. On the basis of previous research, this study separated steroidal saponins to further clarify the chemical composition of the aerial parts of PPC. As a result, three pairs of 25R or 25S epimers of furostanol saponins were obtained by various column chromatography techniques. Their structures were identified as neosolanigroside Y6(1), solanigroside Y6(2), neoprotogracillin(3), protogracillin(4), neoprotodioscin(5) and protodioscin(6) by spectral data combining with chemical transformation. Compound 1 is a new compound, and compounds 2, 3 and 5 are isolated from Paris plants for the first time. Compounds 4 and 6 are isolated from this plant for the first time. Previously, only several spirostanol glycosides with 25S configuration were isolated from Paris plants. Guided by mass spectrometry, the present study isolated the furostanol saponins with 25S configuration from this genus for the first time, which further enriches the chemical information of Paris genus and provides a reference for the isolation of similar compounds.

Qualitative and quantitative analysis of Paris polyphylla var. chinensis by UPLC-Q-TOF-MS/MS and HPLC / 中国中药杂志

Paridis Rhizoma(PR) is prepared from the dried rhizome of Paris polyphylla var. yunnanensis(PPY) or P. polyphylla var. chinensis(PPC) in Liliaceae family. The rapid development of PPY or PPC planting industry resulted from resource shortage has caused the waste of a large number of non-medicinal resources. To clarify the chemical compositions in rhizomes, fibrous roots, stems, leaves, seeds and pericarps of PPC, and explore the comprehensive application value and development prospect of these parts, the qualitative and quantitative analyses on the different parts of PPC were carried out by ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) and high performance liquid chromatography(HPLC). A total of 136 compounds were identified, including 112 steroidal saponins, 6 flavonoids, 11 nitrogen-containing compounds and 7 phytosterols. Rhizomes, fibrous roots, and seeds mainly contained protopennogenyl glycosides and pennogenyl glycosides; leaves and stems mainly contained protodiosgenyl glycosides and diosgenyl glycosides; pericarps mainly contained pennogenyl glycosides, followed by diosgenyl glycosides. The total level of four saponins was the highest in fibrous roots and rhizomes, followed by those in the pericarps and arillate seeds, and the lowest in the stems and exarillate seeds. This study can provide data support for the comprehensive development and rational application of non-medicinal parts of PPC.

Spectrum-effect Relationship of Anti-hepatoma Activity from 3 Kinds of Genus Paris Based on Cluster Analysis and Grey Relational Analysis / 中国实验方剂学杂志

Objective::To study the spectrum-effect relationship between HPLC fingerprints and anti-hepatoma activity of Paris polyphylla var. yunnanensis, P. forrestii and P. vietnamensis, and to elucidate its effective substance. Method::HPLC was used to establish the fingerprint of three extracts from the plant. The mobile phase was consisted of acetonitrile (A)-water (B) for gradient elution (0-10 min, 20%A; 10-20 min, 20%-25%A; 20-30 min, 25%-30%A; 30-40 min, 30%-35%A; 40-50 min, 35%-40%A; 50-60 min, 40%A; 60-75 min, 40%-45%A; 75-80 min, 45%-60%A), and the flow rate was 0.9 mL·min-1. The UV detection wavelength was 203 nm. Thiazolyl blue tetrazolium bromide (MTT) array was used to detect the inhibitory effects of three extracts on the proliferation of HepG2 cells, and the half inhibitory concentration (IC50) was calculated. Cluster analysis and grey relational analysis were used to analyze the data of spectrum and efficacy, and to find out the components that contributed a lot to the anti-liver cancer effect. Result::A total of 11 common peaks were identified as common peaks among HPLC fingerprints of three kinds of Paris. After treated 72 h, P. forrestii has the highest inhibitory effect on the HepG2 cells, the IC50 of P. forrestii, P. polyphylla var. yunnanensis and P. vietnamensis were 148.33, 178.87, 208.09 mg·L-1, respectively. According to the grey relational analysis, the common peaks 1-10 from P. polyphylla var. yunnanensis had great correlation to anti-tumor effect, and the common peaks 1-7 for P. forrestii, the common peaks 1-4, 6-10, N1 for P. vietnamensis, all the correlation degrees with IC50 were >0.7.Cluster analysis of variables in each Paris showed that peaks with correlation degree >0.7 could cluster with IC50. Conclusion::The established HPLC fingerprint method is reliable with good reproducibility. The peaks 1-4, 6 and 7 from three kinds of Paris have the greatest contribution to the anti-hepatoma effect.

A study on quality of Paridis Rhizoma and consideration on standards of Paridis Rhizoma in Chinese Pharmacopoeia / 中国中药杂志

Thin layer chromatography, high performance liquid chromatography and multivariate statistical analysis were integrated in current study to provide a basis for the quality evaluation and the standard improvement of Paridis Rhizoma(Chinese name: Chong-lou). The results demonstrated that the primary saponins in the two authorized sources of Paridis Rhizoma were polyphyllinsⅠ, Ⅱ and Ⅶ, while the rhizome of Trillium tschonoskii an adulterant of Paridis Rhizoma was rich of polyphyllin Ⅵ. Therefore, the apparent content of polyphyllin Ⅵ plays a determinant role towards the source authentication of raw materials and decoction slices of Paridis Rhizoma, whose adulterants frequently occur in the market. Moreover, the contents of polyphyllin Ⅵ in the two authorized sources could meet the requirements of Chinese Pharmacopoeia. Therefore, we suggested that polyphyllin Ⅵ should not be omitted from the quality standard of Paridis Rhizoma in the Chinese Pharmacopoeia, and on the other side, polyphyllinsⅠ, Ⅱ and Ⅶ should be the eligible quality indicators. The study aims to sound information and evidences for the quality evaluation of Paridis Rhizoma, and also to provide a theoretical basis for the standard revision of Paridis Rhizoma in the future Chinese Pharmacopoeia.

Diversity and biomarker exploration of endophytic fungi communities in different varieties of Paridis Rhizoma / 中草药

Objective: The analysis of endophytic fungi diversity and the biomarkers in different varieties of Paridis Rhizoma were used to lay a foundation for the development of quality equivalence and identification and traceability technology. Methods: The amplicons of endophytic fungal community genes from three varieties of Paridis Rhizomaa were sequenced by using Illmina Hiseq high-throughput sequencing. Bioinformatic analysis was performed to study the diversity of flora and the difference of fungal community. Results: The α diversity of endophytic fungi from different varieties was different, but there was no significant difference in β diversity. The OTU of Paris fargesii var. petiolata (Baker ex C. H. Wright) in endophytic fungi was only 0.27%. The results of fungal community annotation showed that the three varieties were annotated to six identical phyla, but the composition ratios were different. Moreover, we explored the biomarker differences in three variants, and the Rhizophagus irregularis of Paris polyphylla through LEfSe analysis. is a biomarker with significant difference. Conclusion: The diversity change of endophytic fungi community of the three varieties was not significant, but the composition ratio was different. The results of this study provided the research basis for the development of quality equivalence and identification and traceability technology.

Industrialization condition and development strategy of Paridis Rhizoma / 中草药

As an important Chinese medicinal material in China, Paridis Rhizoma has obvious curative effect and has analgesic, hemostasis, anti-inflammatory and anti-tumor activities. At present, it is the main raw material of 262 kinds of Chinese patent medicines such as Yunnan Baiyao and Gongxuening Capsule, and has important economic and social value in the Chinese medicine industry. The author mainly systematically summarizes the medicinal history and origin changes of Paridis Rhizoma, the history of industrial development, the status quo of ethnic and folk applications, the status quo of development and application of proprietary Chinese medicines, and the study of resource ecology and breeding, and proposes the implementation of the process of industrialization of Paridis Rhizoma. Important issues and countermeasures need to be paid attention in order to provide reference for the sound development of Paridis Rhizoma.

UPLC quantitative analysis of Paridis Rhizoma and its comprehensive evaluation of chemical quality / 中草药

Objective: To explore the content of seven active compounds in 10 kinds of medicinal herbs of Paridis Rhizoma, and to carry out chemical composition integration evaluation, which provides a scientific basis for its resource utilization. Methods: A total of 55 batches of medicinal herbs were collected from different areas, and their saponins I, saponins II, saponins VI, saponins VII, diosgenin, saponins H, and saponins were determined by ultra performance liquid chromatography. Then, TOPSIS model was used to normalize the content results and integrate the multi-indicator data to obtain a comprehensive index of content of seven active compounds. Results: The 10 kinds of medicinal herbs of Paridis Rhizoma were ranked from long to low was Paris forrestii (Ci = 0.275 5) > Paris polyphylla (Ci = 0.273 2) > Paris daliensis (Ci = 0.269 8) > Paris polyphylla var. yunnanensis (Ci = 0.244 5) > Paris vietnamensis (Ci = 0.234 5) > Paris polyphylla var. stenophylla (Ci = 0.159 1) > Paris thibetica (Ci = 0.141 6) > Paris polyphylla var. nana (Ci = 0.117 8) > P. vietnamensis (Ci = 0.115 1) > Paris mairei (Ci = 0.114 9), indicating that comprehensive quality of 10 kinds of medicinal herbs of Paridis Rhizoma had a large gap. The overall quality of P. forrestii, P. polyphylla and P. daliensis are better than that of P. polyphylla var. yunnanensis, and the comprehensive evaluation results of P. vietnamensis and P. polyphylla var. yunnanensis are closer that can be used as an alternative species for resource mining. Conclusion: The comprehensive evaluation of chemical quality has certain reference value for the quality evaluation of Paridis Rhizoma

Determination and Analysis of Heavy Metals of Paridis Rhizoma from Different Localities and Pieces / 中国实验方剂学杂志

Objective:To analyze and determine heavy metal content in Paridis Rhizoma from different genus and localities,in order to provide a reference for selecting cultivation areas and establishing the quality standard of Paridis Rhizoma of heavy metals content. Method:Microwave digestion method combined with inductively coupled plasma atomic emission spectrometry method (ICP-AES) method were applied to determine the contents of 6 heavy metals,i.e. As,Cu,Hg,Cd,Pb and Cr in 39 samples of Paridis Rhizoma of different genus and localities in Yunnan Province. Cluster analysis,statistical analysis and principal component analysis (PCA) were used to compare the differences of heavy metals contents in different localities and species. Result:The contents of six heavy metals in Paridis Rhizoma met the ISO international standard of Heavy Metal Limit of Traditional Chinese Medicine-traditional Chinese Medicinal Materials. Under the limited value standard of Green Trade Standards of Importing Medicinal Plants and Preparations,the over-standard rate of heavy metal As was 15.4%,the excess rate of Cd was 5.1%,and the excess rate of Pb was 2.6%. The contents of Cu and Hg conformed to relevant requirements. Cluster analysis,statistical analysis and principal component analysis showed that for the same variety,differences in producing places had significant effects on heavy metal content,while differences in species had little effects. Conclusion:The results of this study indicated that the heavy metal content of Paris planted in and around Dali basically conformed to relevant standards. The differences of heavy metal content in Paris were mainly regional differences,which provided a theoretical basis for standardizing the cultivation of medicinal materials and formulating the limit standards of heavy metals for Paridis Rhizoma.

Improving Quality Standard of Processed Slices of Paridis Rhizoma of Chinese Pharmacopoeia / 中国实验方剂学杂志

Objective:To study the appearance description,TLC examination and content determination was carried out, in order to improve the quality standard of processed slices of Paridis Rhizoma in the 2020 edition of Chinese Pharmacopoeia. Method:Based on the literature review and observation on the samples,the appearance description was described. TLC examination was used for the qualitative analysis. HPLC was used for the determination of polyphyllin Ⅰ,Ⅱ,Ⅵ and Ⅶ in the commercial and processed samples. UPLC was employed for the determination of 10 steroidal saponins,namely pseudoprotodioscin,polyphyllin Ⅶ,17-hydroxygracillin,polyphyllin H,polyphyllin Ⅵ,polyphyllin Ⅱ,dioscin,gracilin,polyphyllin Ⅰ and polyphyllin Ⅴ. Result:For the appearance description,color and luster,texture,odor and taste as well as the diameter of 1.0-4.5 cm were recorded. polyphyllin Ⅵ was not detected in the thin layer chromatograms of most of the tested samples derived from high-quality species but obviously detected in those of Trillium Rhizoma. Five of 13 commercial samples met the requirements that the total amounts of polyphyllin Ⅶ,Ⅵ,Ⅱ,and Ⅰ should be no less than 0.6%according to the current Chinese Pharmacopoeia. Because softening and drying had the obvious influence on the contents of steroidal saponins in the samples,soaking and sun-drying were preferred. Conclusion:Appearance description should be supplemented. Polyphyllin Ⅵ is not considered as one of quality markers for the TLC identification and HPLC determination of Paridis Rhizoma. Polyphyllin H was considered as a new marker for the quality control. It is recommended that the total amounts of polyphyllin Ⅶ,H,Ⅱ,and Ⅰ should be no less than 1.0%.

A screening method and practice for alternative resources of Paridis Rhizoma based on overall similarity of components and activities / 中草药

Objective To study the overall similarity of composition-activity of different kinds of Paridis Rhizoma, and establish a new method for selecting alternative resources. Methods The HPLC method was used to detect the content of polyphyllin I, II, VI, and VII, and pharmacological model of analgesic and hemostatic were used to detect the efficacy. The data were standardized using standard deviation method. Using polyphyllin I, II, VI, and VII at minimum 0.6%, aspirin and Yunnan Baiyao Group analgesic and hemostatic efficacy data as standard controls, we also established component and activity indexes. SPSS 20.0 Software was used to analyze the principal component analysis (PCA) and hierarchical cluster analysis (HCA) of these seven medicinal plants. Results The total content of the main saponins in seven medicinal plants of Paris genus ranged from 0.125% to 1.649%, and the maximum content (1.649%) was found in the P. forrestii while the minimum content (0.125%) was found in P. daliensis. The study also disclosed that P. polyphylla var. yunnanensis has the most active analgesic activity, and the time of bleeding (BT) in mice and the time of coagulation (CT) in mice were the shortest in P. polyphylla. The shortest mice activated partial thrombin time (APTT) was P. forrestii and the longest was P. thibetica. The shortest prothrombin time (PT) in mice was P. vietnamensis. The longest BT in mice was P. daliensis. The longest CT and PT in mice was P. polyphylla. PCA and HCA results showed that P. polyphylla var. yunnanensis, P. vietnamensis and P. thibetica were in the same class, while P. pubescens, P. daliensis, P. polyphylla, and P. forrestii were in another class. It is indicated that the overall similarity of component-activity of P. vietnamensis, P. thibetica, and P. polyphylla var. yunnanensis was higher, thus they can be considered as alternative sources. Conclusion The quality relations of different kinds of Paris genus were evaluated by the overall similarity of the component and activity index, which provides ideas and methods for the search of the replacement resources of the endangered plants.

Variation of Rhizospheric Microorganisms and Soil Enzyme Activity of Paridis Rhizoma Cultivated in Three Gorges Reservoir Region / 中国中医药信息杂志

Objective To study the amount of rhizospheric microorganisms and soil enzyme activity influenced by Paridis Rhizoma in different locations and of different strains. Methods The amount of rhizospheric microorganisms, soil enzyme activity and their correlation were researched through field survey and collection of rhizospheric soil in Paridis Rhizoma cultivated in Three Gorges Reservoir Region and by microbial dilution plate culture method. Results The amount of rhizospheric microorganisms in Paridis Rhizoma from different habitats showed significant differences. The dominant species in soil microflora was bacteria; the second one was actinomycetes; the fewest one was fungus. The variation trend of the amount of rhizospheric microorganisms was not consistent with the variation trend of rhizospheric microorganisms diversity index. The activity of soil phosphatase, invertase and pepsin in Paridis Rhizoma from different habitats varied. The correlation analysis showed that the correlation between the soil enzyme activity and the amount of rhizospheric microorganisms existed. Conclusion Choosing the suitable strains and habitats of Paridis Rhizoma is beneficial to enhancing the amount of rhizospheric microorganisms and soil enzyme activity, which can create good micro-ecological environment for growth and cultivation of Paridis Rhizoma.

Correlation of Active Ingredients in the Roots of Paridis rhizoma Genus Plants with Mycorrhizal Infection Rate and Soil Nutrients / 中国药房

OBJECTIVE:To investigate the correlation of total saponins and total polysaccharides content in the roots of Pari-dis rhizoma genus plants with Arbuscular mycorrhizal fungi(AMF)infection rate and rhizosphere soil nutrients. METHODS:Tak-ing 16 pieces of Paridis rhizoma herbs and rhizosphere soil in Three Gorges Reservoir Area as the research object,mycorrhizal in-fection rate and infection intensity,the contents of total saponins and total polysaccharides in roots and stem,and rhizosphere soil nutrient contents were all determined. SPSS18.0 software was adopted for data multiple comparison and correlation analysis. RE-SULTS:Paridis rhizoma genus plants roots in Three Gorges Reservoir Area could form a good symbiotic relationship with AMF. In-fection rate ranged from 85.19% to 99.80%,and infection intensity ranged from 16.55% to 72.27%. Paridis rhizoma from differ-ent origins and varieties had significant difference in the contents of total saponins and total polysaccharides (P<0.05). Soil pH was moderate,the average content of total nitrogen was in insufficient level,the average concentration of the remaining nutrients belonged to middle levels or above. Correlation analysis showed that the mycorrhizal infection rate and infection intensity of Paridis rhizoma were not significantly correlated with its quality. The total saponins were significantly negatively correlated with organic matter and available nitrogen. The total saponins and total polysaccharides were positively correlated with available phosphorus,po-tassium and pH value,but they were not significantly correlated with the other soil nutrients. CONCLUSIONS:Paridis rhizoma is selective to different soil nutrients. Rich soil available phosphorus and potassium supply are in favor of the synthesis and accumula-tion of total saponins and total polysaccharides in Paridis rhizoma roots.

Patent Information Analysis of Cultivation and Seed Treatment Techniques of Paridis Rhizoma / 中国药房

OBJECTIVE:To provide reference for the patent application of artificial cultivation and seed treatment technology of Paridis Rhizoma and natural resources protection. METHODS:Retrieved from State Intellectual Property Office patent database,“Paridis Rhizoma”and“Paris polyphylla”were used as search keywords from 1985 to March 31,2015;Paridis Rhizoma cultiva-tion and seed treatment technology patents as research object were analyzed in fields of the amount of patent application,regional construction and legal status,etc. RESULTS:Among 54 Paridis Rhizoma cultivation patent application,there were 19 applications from Yunnan,9 from Sichuan,8 from Hubei,5 from Anhui,4 from Hunan,3 from Guizhou,2 from Jiangxi,1 from Fujian, Shanxi,Jiangsu,Guangxi respectively;there were 18 licensed patents,and 13 licensed patents maintained more than 3 years, among which 1 patent right transferred;from aspect of patent without right,6 patent right were given up due to fail to pay annual fee,1 rejected and 5 withdrawn;from main types of patent proposer,enterprise submitted 24 patent application,universities 15, scientific research institutes 8 and other 7. 24 enterprise patents involved 21 enterprises,indicating that patent protection net didn’t form due to disperse proposers. CONCLUSIONS:Patent application is mainly from Yunnan. There still are some problems,such as small number of patent applications,poor quality of application writing,little patent transformation. It is suggested to plan artificial cultivation technology,enhance the quality of patent application,pay attention to patent operation and achievement transformation and other measures,in order to protect Paridis Rhizoma resource and industrial development.

Content Determination of Diosgenin in Paridis Rhizoma Medicinal Plants in Different Habitats of Three Gorges Reservoir / 中国中医药信息杂志

Objective To establish an HPLC method to determine the content of diosgenin in Paridis Rhizoma; To examine contents of diosgenin in different varieties of Paridis Rhizoma medicinal plants and from different habitats. Methods Shimadzu Inertsil ODS-C18 guard column (150 mm×4.6 mm, 5μm) was used in HPLC system; elution conditions were the mobile phase using 92% acetonitrile and 8% water (isocraticelution) at a flow rate of 1.0 mL/min with 203 nm detection wavelength. The temperature of the column oven was maintained at 30℃.Results Different varieties of Paridis Rhizoma medicinal plants and Paridis Rhizoma medicinal plants from different habitats contained diosgenin. TheParis polyphylla var.pseudothibetica in Yubei district had the highest content (6.813 7 mg/g);Paris polyphylla var.stenophylla in Chengkou County had the middle content (5.758 4 mg/g);Paris yunnanensisFranch in Shizhu county showed the lowest content (1.952 2 mg/g). The results showed that the obvious difference of chemical quality between various cultivated varieties of Paridis Rhizoma medicinal plants, which might be related with origin, variety and the cultivation technology, and so on.Conclusion The diosgenin contents ofParis polyphylla var. pseudothibetica andParis polyphylla var.stenophylla were higher than that of varieties recorded in Chinese Pharmacopeia.Paris polyphylla var.pseudothibetica andParis polyphylla var.stenophylla had important medical value.

Research Progress in Seeding Breeding of Paridis Rhizoma / 中国中医药信息杂志

Paridis Rhizoma is a rare Chinese herbal medicine with variety of medicinal values. In recent years, the demand for Paridis Rhizoma has increased gradually. Artificial cultivation has met difficulties, while exploitation of wild resources was caught in a vicious circle and has overdrawn seriously. So it is of great significance to enhance the protection of Paridis Rhizoma, carry out basic research, in order to solve problems in seeding breeding, promote artificial cultivation to meet the market need and achieve sustainable development and supply. This article reviewed the status qua of seedling breeding of Paridis Rhizoma, including seed breeding, tuber breeding and tissue culture, with a purpose to standardize planting of resource conservation and utilization of Paridis Rhizoma.

Preparation and in vitro release evaluation of pH-dependent colon targetting pellets of Paridis Rhizoma saponin and Astragali Radix polysaccharide / 中草药

Objective: To prepare pH-dependent Paridis Rhizoma saponin (PRS) and Astragali Radix polysaccharide (ARP) colon targeting pellets for the treatment of colon cancer and finish its in vitro release performance evaluation. Methods: The colon targeted pellets were prepared with extrusion-spheronization and air-flow coating method and the the process parameters were optimized by orthogonal design. The coating fluid prescription was investigated by single factor test. In vitro release performance evaluation of the pellets was evaluated with polyphyllin I and II as the indexes. Results: The optimum technologic parameters of extrusion spheronization equipment were as follows: the rate of extrusion was 60 r/min, the rate of spheronization was 1 200 r/min, and the time of spheronization was 5 min. The optimum coating liquid formulation of pH-dependent colon targetting pellets was 15% weight gains of Eudrugit S100, 1.5% anti-plastering aid amount of Glycerin monostearate, and 5% plasticizer amount of TEC. In vitro release test showed that cumulative release rate of berberine hydrochloride was close to 0% in artificial gastric juice after 2 h and less than 10% in artificial intestinal fluid after 4 h, but the cumulative release rate in artificial colon juice after 2 h was more than 90%. Conclusion: The preparation method can be applied to the preparation of colon targeted pellets and the pellets can achieve the targeted release in the colon.

Determination of seven steroidal saponins in Paridis Rhizoma and polygerm varieties from different regions in Yunnan province by UPLC and establishment of fingerprint / 中草药

Objective: To establish a quantitative analysis method of seven steroidal saponins in Paridis Rhizoma and its polygerm varieties from different regions in Yunnan province, to simultaneously determine the contents of seven steroidal saponins, to establish the fingerprint for the polygerm varieties in Gaoligong Mountain by UPLC-ELSD, and to evaluate the qualities. Methods: It was detected with an Acquity UPLC® BEH C18 (2.1 mm × 50 mm, 1.7 μm) column, and gradually diluted with acetonitrile-water at a flow rate of 0.2 mL/min, drift tube temperature of 55℃, nitrogen pressure 275.8 kPa, and gain 500 by UPLC-ELSD. Results: The 23 samples could be separated and analyzed of seven steroids within 23 min. All the indexes of the methodological investigation met the requirements. The results showed that the contents of saponins from 19 samples met the requirement in Chinese Pharmacopoeia 2015. The average content of four saponins was 1.42% and among them the content from the sample in Gaoligong Mounta was up to 1.660%. The similarity was lower than 0.9 in the samples of Gaoligong Mountain, and there were eight common peaks in the fingerprints, which were identified four characteristic peaks. The type and content of seven saponins in the samples could not be clearly classified by PCA analysis and system cluster analysis. Conclusion: The method is convenient, accurate, and suitable for the quantitative analysis of Paridis Rhizoma and polygerm varieties. There is no obvious difference between Paridis Rhizoma and polygerm varieties in chemical components and contents. Paridis Rhizoma (polygerm varieties) in Gaoligong Mountain is a variety worth popularization and in-depth researches.