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ObjectiveIn recent years， with the sharp decline of wild resources in Arisaematis Rhizoma and Pinelliae Rhizoma and the immaturity of medicinal cultivation technology， their adulterants have appeared frequently in the market， and the main identifying characteristics have mostly disappeared in the circulation of medicinal materials. Therefore， there is an urgent need to establish a molecular identification method that can quickly and effectively identify the specificity of Arisaematis Rhizoma and Pinelliae Rhizoma. MethodAfter comparison of the rbcL sequences of Arisaematis Rhizoma，Pinelliae Rhizoma， and their adulterants， the specific enzyme cleavage sites Hae Ⅲ and Dra Ⅰ of Arisaematis Rhizoma and Pinelliae Rhizoma， respectively， were selected and identified by polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP）. The main system conditions of PCR-RFLP reaction were established and optimized， and their durability and the ability to detect genuine， adulterants， and mixed counterfeits were investigated. ResultThe PCR-RFLP identification method of Arisaematis Rhizoma and Pinelliae Rhizoma was established. After specific primer amplification， Arisaematis Rhizoma and Pinelliae Rhizoma could be digested by Hae Ⅲ and Dra Ⅰ-restricted endonucleases respectively， at annealing temperature of 54 ℃， the number of cycles of 35， and the amount of DNA template of 3-30 ng， producing two fragments or small cut fragments with a single band between 100-250 bp， whereas the mixed counterfeits were not cleaved and both showed a band at 250 bp. The method is highly accurate in identifying adulterants and mixed counterfeits of Arisaematis Rhizoma or Pinelliae Rhizoma. ConclusionThe PCR-RFLP method developed in this study allows for the rapid identification of Arisaematis Rhizoma and Pinelliae Rhizoma.
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The present study investigated the effect of immersion in the excipient lime water on the toxic component lectin protein and explained the scientific connotation of lime water detoxication during the processing of Pinelliae Rhizoma Praeparatum. Western blot was used to investigate the effects of immersion in lime water with different pH(pH 10, 11, and 12.4), saturated sodium hydroxide, and sodium bicarbonate solution on the content of lectin protein. The protein compositions of the supernatant and the precipitate after immersing lectin protein in lime water of different pH were determined by the SDS-PAGE method combined with the silver staining technique. The MALDI-TOF-MS/MS technique was used to detect the molecular weight distribution of peptide fragments in the supernatant and precipitate after immersing lectin protein in lime water of different pH, and circular dichroism spectroscopy was used to detect the ratio changes in the secondary structure of lectin protein during the immersion. The results showed that immersion in lime water at pH>12 and saturated sodium hydroxide solution could significantly reduce the content of lectin protein, while immersion in lime water at pH<12 and sodium bicarbonate solution had no significant effect on lectin protein content. The corresponding lectin protein bands and molecular ion peaks were not detected at the 12 kDa position in the supernatant and precipitate after immersing the lectin protein in lime water at pH>12, which was attributed to the fact that lime water immersion at pH>12 could significantly change the ratio of the secondary structure of lectin protein, resulting in irreversible denaturation, while lime water immersion at pH<12 did not change the ratio of the secondary structure of lectin protein. Therefore, pH>12 was the key condition for the detoxication of lime water during the processing of Pinelliae Rhizoma Praeparatum. Lime water immersion at pH>12 could cause irreversible denaturation of lectin protein, resulting in a significant decrease in the inflammatory toxicity of Pinelliae Rhizoma Praeparatum, which played a key role in detoxification.
Subject(s)
Lectins , Pinellia , Sodium Bicarbonate , Sodium Hydroxide , Tandem Mass Spectrometry , WaterABSTRACT
ObjectiveTo analyze and summarize the medication rules of different Pinelliae Rhizoma processed products in the syndrome differentiation and treatment of insomnia using data mining. MethodThe literature on the treatment of insomnia with Pinelliae Rhizoma was retrieved from the China National Knowledge Infrastructure （CNKI）， VIP， and PubMed databases over the past 10 years. An Excel database was constructed to record the prescriptions of different Pinelliae Rhizoma processed products in the treatment of insomnia. SPSS 26.0 software was used for frequency analysis of traditional Chinese medicine （TCM） syndromes related to insomnia， compatibility of drugs， drug effects， and properties. SPSS 26.0 was also used for cluster analysis， factor analysis， and IBM Modeler 18.0 plugin for association rule analysis of the core compatibility of different Pinelliae Rhizoma processed products and combinations. ResultAfter applying inclusion and exclusion criteria， 125 relevant articles were finally included. The commonly used processed products of Pinelliae Rhizoma in the treatment of insomnia were Pinelliae Rhizoma Praeparatum， Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine， and Pinelliae Rhizoma Praeparatum cum Alumine. Among them， Pinelliae Rhizoma Praeparatum was the most frequently used. All three processed products of Pinelliae Rhizoma were often used for insomnia with such TCM syndromes as phlegm-heat disturbing the heart， phlegm-dampness obstructing the interior， and liver Qi stagnation. The compatible drugs were sweet， bitter， and pungent in flavor， cold in nature， and acted on the lung， spleen， heart， and liver meridians， with functions of nourishing deficiency， clearing heat， and calming the mind. The common prescriptions used were Wendantang， Chaihu Longgu Mulitang， Banxia Xiexintang， and Xiaochaihutang， with doses ranging from 6 to 30 g. The core drug combinations were Pinelliae Rhizoma Praeparatum-Poria-Ziziphi Spinosae Semen， Pinelliae Rhizoma Praeparatum Cum Zingibere et Alumine-Jujubae Fructus-Codonopsis Radix， and Pinelliae Rhizoma Praeparatum Cum Alumine-Scutellariae Radix-Bupleuri Radix. ConclusionThis study， for the first time， analyzed and summarized the compatibility and prescription application rules of commonly used processed products of Pinelliae Rhizoma in the treatment of insomnia from the perspective of TCM syndrome differentiation， which provides a theoretical basis for the rational， safe， and effective use of Pinelliae Rhizoma in the treatment of insomnia in TCM.
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In this paper， by consulting the historical herbs and medical classics coupled with related literature in modern research， the historical edition of Chinese Pharmacopoeia and local chronicles of traditional Chinese medicine （TCM） along with the ancient historical evolution of the processing methods of Pinelliae Rhizoma origin as well as the related processing methods of Pinelliae Rhizoma origin from 1959 to 2020 were systematically collated and summarized. It was found that the main processing methods of Pinelliae Rhizoma origin were peeling， decoction washing， lime wrapping and sun-drying. However， stacking， peeling， sun-drying or oven-drying are the primary methods in modern local chronicles of TCM. Meanwhile， washing， peeling， removing fibrous roots and sun-drying are the main methods in Chinese Pharmacopoeia. In addition， there were some changes in the quality evaluation of Pinelliae Rhizoma in different historical periods. Round and white were the best in the quality evaluation of Pinelliae Rhizoma in ancient times， while the evaluation indexes were further refined to size， color， texture， powder property， purity and evenness in modern herbal works. In modern studies， the quality of Pinelliae Rhizoma was mostly evaluated by the chemical components such as alkaloids， total organic acids， polysaccharides， nucleosides， fingerprint and pharmacodynamics. At present， the purification and drying stages of Pinelliae Rhizoma are in the transitional stage between the traditional manual peeling and natural drying methods as well as the modern mechanized and large-scale production. Therefore， a reasonable and feasible modern processing methods and guiding standards of Pinelliae Rhizoma are developed urgently to normalize the processing of Pinelliae Rhizoma and ensure the quality of medicinal materials.
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In this paper， by consulting the historical herbs and medical classics coupled with related literature in modern research， the historical edition of Chinese Pharmacopoeia and local chronicles of traditional Chinese medicine （TCM） along with the ancient historical evolution of the processing methods of Pinelliae Rhizoma origin as well as the related processing methods of Pinelliae Rhizoma origin from 1959 to 2020 were systematically collated and summarized. It was found that the main processing methods of Pinelliae Rhizoma origin were peeling， decoction washing， lime wrapping and sun-drying. However， stacking， peeling， sun-drying or oven-drying are the primary methods in modern local chronicles of TCM. Meanwhile， washing， peeling， removing fibrous roots and sun-drying are the main methods in Chinese Pharmacopoeia. In addition， there were some changes in the quality evaluation of Pinelliae Rhizoma in different historical periods. Round and white were the best in the quality evaluation of Pinelliae Rhizoma in ancient times， while the evaluation indexes were further refined to size， color， texture， powder property， purity and evenness in modern herbal works. In modern studies， the quality of Pinelliae Rhizoma was mostly evaluated by the chemical components such as alkaloids， total organic acids， polysaccharides， nucleosides， fingerprint and pharmacodynamics. At present， the purification and drying stages of Pinelliae Rhizoma are in the transitional stage between the traditional manual peeling and natural drying methods as well as the modern mechanized and large-scale production. Therefore， a reasonable and feasible modern processing methods and guiding standards of Pinelliae Rhizoma are developed urgently to normalize the processing of Pinelliae Rhizoma and ensure the quality of medicinal materials.
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This study investigated the material basis and mechanism of Pinelliae Rhizoma Decoction in the treatment of airway inflammation. The cigarette smoke combined with lipopolysaccharide(LPS) was used to induce an airway inflammation model in mice. The expression levels of IL-6 and IL-8 in the bronchoalveolar lavage fluid(BALF) and the phosphorylation levels of p38 and IκB in the lungs of mice were taken as indexes to screen the effective extracts by system solvent extraction from Pinelliae Rhizoma Decoction(dichloromethane extract, ethyl acetate extract, n-butanol extract, etc.). Meanwhile, the human bronchial epithelial(16-HBE) cell model of cigarette smoke extract(CSE)-induced injury was established, and the mRNA expression levels of IL-6 and IL-8 and the phosphorylation levels of p38 and IκB proteins were also taken as indexes to evaluate the anti-inflammatory effect of different extracts of Pinelliae Rhizoma Decoction. The results showed that Pinelliae Rhizoma Decoction significantly antagonized airway inflammation in mice by down-regulating the expression levels of IL-6 and IL-8 in mice with airway inflammation and 16-HBE cells with CSE-induced injury and inhibiting the phosphorylation levels of p38 and IκB. The dichloromethane and ethyl acetate extracts of Pinelliae Rhizoma Decoction showed significant anti-inflammatory effects, while such effects of other extracts were not prominent. Furthermore, the database of Pinelliae Rhizoma composition was constructed, and the components in effective extracts were analyzed by HPLC-TOF-MS and Nano-LC-MS/MS. As revealed by the results, the compositions of the two effective extracts were similar with 36 common components. They were combined and then divided into Pinelliae Rhizoma alkaloids(PTAs) and Pinelliae Rhizoma non-alkaloids(PTNAs) by 732 cation-exchange resin. Further in vitro investigation confirmed the significant anti-inflammatory effect of PTNAs, while such effect of PTAs was not manifest. The MS analysis showed 172 peptides and 7 organic acids in PTNAs. The peptide content in PTNAs was 63.5% measured by quantitative analysis of BCA assay, and the organic acid content was 9.92% by potentiometric titration method. The findings of this study suggested that Pinelliae Rhizoma Decoction could antagonize airway inflammation in mice by inhibiting phosphorylation of p38 and IκB and blocking the activation of MAPK and NF-κB signaling pathways, and the effective components were related to the peptides and organic acids in PTNAs. The above results lay a foundation for the research on the mechanism and material basis of Pinelliae Rhizoma in antagonizing airway inflammation.
Subject(s)
Animals , Mice , Drugs, Chinese Herbal/pharmacology , Inflammation/drug therapy , NF-kappa B/genetics , Pinellia/chemistry , Respiratory Tract Diseases/drug therapy , Rhizome , Tandem Mass SpectrometryABSTRACT
Objective:To evaluate the scientificity and feasibility of processing of Pinelliae Rhizoma by hot water washing （Tangxi）， and to provide reference for the development of related famous classical formulas. Method:Processing method of Pinelliae Rhizoma washed by hot water was established based on ancient Tangxi processing method， and the process conditions were optimized by single factor tests. The weight， moisture， ash， extract， total acid （calculated by succinic acid） contents and high performance liquid chromatography （HPLC） fingerprint of Pinelliae Rhizoma were compared before and after processing. In addition， the rabbit eye irritation test was conducted to evaluate the toxicity changes. Result:The processing method of Pinelliae Rhizoma washed by hot water was as following：washed by 4 times the amount of hot water at 80 ℃ for 10 times until clear water， transfused cross-section after incision， no or slight numbness in the mouth. The average moisture， ash， extract contents of Pinelliae Rhizoma washed by hot water were 9.34%， 1.71% and 4.22%， respectively. After being processed， the decline rates of weight and total acid content of Pinelliae Rhizoma were 7.49% and 43.31%. The HPLC fingerprint of Pinelliae Rhizoma before and after washing showed a decrease in all components， but there was no new chromatographic peak， and peak 9 （adenosine） reduced significantly. The results of rabbit eye irritation test showed that there was no obvious eye conjunctival irritation after washing， indicating that the toxicity of Pinelliae Rhizoma decreased obviously after washing. Conclusion:The established method of Pinelliae Rhizoma by Tangxi processing is stable and feasible， the aqueous extract of Pinelliae Rhizoma has no obvious eye conjunctival irritation after washing.
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There are 17 formulas containing Pinelliae Rhizoma in the Catalogue of Ancient Famous Classical Formulas (the First Batch), most of which are only labeled with " washing with decoction" or without any processing method, which is inconsistent with the current use requirements and brings confusion to research and development of related famous classical formulas. Through combing the records of famous classical formulas in the original book, contemporary works and later works, the usage of processed products of Pinelliae Rhizoma in the evolution of past dynasties was contrasted, and the efficacy, indications and compatibility significance of different formulas were analyzed, according to the principle of " respecting the ancient but not confining the ancient" , the authors suggested that the 17 famous classical formulas should use the processed products of Pinelliae Rhizoma in the 2015 edition of Chinese Pharmacopoeia, including Pinelliae Rhizoma Praeparatum, Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine and Pinelliae Rhizoma Praeparatum cum Alumine. Among them, Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine should be used in Banxia Xiexintang, Gancao Xiexintang, Huangliantang, Xuanfu Daizhe Tang, Zhuye Shigaotang, Banxia Houpotang, Maimendong Tang, Gualou Xiebai Banxiatang, Wendantang, Zhurutang, Jinshui Liujun Jian and Yangweitang, Pinelliae Rhizoma Praeparatum should be used in Shengyang Yiweitang, Houpo Mahuangtang, Banxia Baizhu Tianma Tang and Huopo Xialing Tang, and Pinelliae Rhizoma Praeparatum cum Alumine should be used in Sangbaipi Tang.
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The present study was aimed to investigate the effect of lime and licorice processing of Pinelliae Rhizoma on its toxic lectin protein and clarify the scientific detoxification connotation of lime and licorice processing of Pinelliae Rhizoma. Western blot was used to semi-quantitatively analyze the contents of lectin in Pinelliae Rhizoma and Pinelliae Rhizoma Praeparatum. Raw products and lectin were treated by soaking in licorice juice, lime solution or mixture solution of these two to investigate the different processing time on the content of toxic lectin protein. SDS-PAGE gel electrophoresis was used to analyze the changes of lectin protein bands in the solution and precipitates before and after processing. MALDI-TOF technology was used to qualitatively analyze and compare the protein molecular weight before and after processing. The results showed that the contents of lectin in Pinelliae Rhizoma and Pinelliae Rhizoma Praeparatum were 5.01% and 0.04% respectively, indicating that processing could significantly reduce the content of active lectin in raw products. The results also showed that the content of lectin in raw drugs decreased significantly after soaking in lime solution for one day or in licorice juice for three day, and the effect was greatest in mixture solution. Qualitative analysis showed that after being treated by soaking in lime solution, the lectin protein was decomposed into small peptide segments, while after being treated by soaking in licorice juice, the lectin protein was denatured and precipitated. The structure of lectin protein in Pinelliae Rhizoma was broken after being processed with licorice juice and lime solution, which significantly reduced the content of toxic lectinprotein. This is one of the detoxification mechanisms of Pinelliae Rhizoma processing.
Subject(s)
Calcium Compounds , Drugs, Chinese Herbal , Glycyrrhiza , Lectins , Oxides , Pinellia , Technology, PharmaceuticalABSTRACT
ABSTRACT OBJECTIVE：To investigate the quality differences of Pinelliae Rhizoma with different diameter，and to providereference for the rational utilization of its resources. METHODS：A total of 65 batches of commercial Pinelliae Rhizoma from thefour major medicinal markets in China and two companies were collected as samples，the diameter of them were measured byvernier caliper，and then them were screened into samples with different diameter ranges（0.5 cm≤d≤0.8 cm，0.8 cm＜d≤1.0cm，1.0 cm＜d≤1.2 cm，1.2 cm＜d≤1.5 cm，1.5 cm＜d≤2.0 cm）by sieving. The content of organic acids（oxalic acid，citricacid，L-malic acid，succinic acid，fumaric acid，trans-aconitic acid and cis-aconitic acid）in Pinelliae Rhizoma samples of differentdiameter ranges were determined by HPLC，and the content of the extracts of Pinelliae Rhizoma samples of different diameterranges were determined by cold leaching method according to 2201 general principles in 2015 edition of Chinese Pharmacopoeia（Part Ⅳ），to evaluate the quality difference of Pinelliae Rhizoma in different diameter ranges.73 batches of Pinelliae Rhizoma fromcorresponding 10 main producing areas were collected as samples，to investigate its distribution of diameter. RESULTS：Dtermination of 65 batches of commercial Pinelliae Rhizoma showed that the content of organic acids and extracts in PinelliaeRhizoma increased with the increase of its diameter in general. However，there were no significant difference in organic acids andcontent of the extracts among Pinelliae Rhizoma samples of different diameter ranges in the same batch（P＞0.05）. The diameter distribution of 73 batches of Pinelliae Rhizoma from corresponding areas was mainly between 0.6 cm and 1.8 cm，with an averageweight of 95.54％. If the diameter standard（1-1.5 cm）specified in the 2015 edition of Chinese Pharmacopoeia（part Ⅰ）wasadopted，only 8 of 73 batches of Pinelliae Rhizoma meet the requirements，and the qualified rate was 10.96％；if the diameterrange was properly expanded to 0.7-1.5 cm，54 batches of Pinelliae Rhizoma meet the requirements，with a qualified rate of73.97％；when the diameter range was expanded to 0.7-1.6 cm，68 batches of Pinelliae Rhizoma meet the requirements，with aqualified rate of 93.15％ .CONCLUSIONS：There is no significant difference in the quality of Pinelliae Rhizoma with different diameters. Combined with the diameter distribution of Pinellia Rhizoma in different producing areas，the diameter range of 1-1.5cm specified in relative standard can be expanded to 0.7-1.6 cm，avoiding the waste of resources.
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Pinelliae Rhizoma is a toxic traditional Chinese medicine, and its dose differs greatly between at ancient and modern times, so that it is difficult for researchers to determine its dosage during research and development of famous classical formulas. According to the literature research, it is found that the dose of Pinelliae Rhizoma in famous classical formulas has undergone a process from large to small. Based on the principle of " following the classic and conforming to pharmacopoeia" , and in combination with the modern actual situation, the reasonable dose of Pinelliae Rhizoma in the 17 formulas from the Catalogue of Ancient Famous Classical Formulas (The First Batch) is determined. According to the records of the original book, the evolution of past dynasties and the consideration of the current clinical application, dosage regimen and administration methods of the 17 famous classical formulas and so on, the authors suggested that the dosages of Pinelliae Rhizoma in Banxia Houpotang and Maimendong Tang are 15.0 g, its dosages in Wendantang and Sangbaipi Tang are 6.0 g, dosages of Pinelliae Rhizoma in Xuanfu Daizhe Tang, Zhuye Shigaotang, Banxia Xiexintang, Gancao Xiexintang, Huangliantang, Gualou Xiebai Banxiatang, Houpo Mahuangtang and Jinshui Liujunjian are 7.5 g, the dosages of Pinelliae Rhizoma in Zhurutang, Shengyang Yiweitang and Yangweitang are 6.7, 4.5, 5.0 g, respectively, dosages of Pinelliae Rhizoma in Banxia Baizhu Tianma Tang and Huopo Xialing Tang are 5.6 g.
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Objective::To establish the evaluation method of Pinelliae Rhizoma (PR) resources by the analytic hierarchy process (AHP) and grey correlation degree (GCD) method, so as to explore the differences in quality components, production efficiency and appearance traits of different germplasm resources cultivated in the same environment. Method::The quality component index, efficiency index, and appearance traits index of 15 germplasm resources were measured, including moisture, total ash, extractives, total acid content, harvest index, yield, drying rate, commodity rate, decay rate and deformability of tubers. Based on AHP and grey correlation method, each indicator data was processed in a comprehensive way, its comprehensive correlation value was calculated, and different PR germplasm resources were comprehensively evaluated. Result::Based on three factors-quality composition, production efficiency and appearance traits, the comprehensive correlation value of A6 was the highest, reaching 0.749 4, which was followed by A14, A15, A7, and their comprehensive correlation values were 0.736 6, 0.726 2, 0.718 2, respectively. Therefore, the source of A6 could be used as an excellent provenance introduced to the cultivation of PR, and the provenance of A14, A15, and A7 could be used as a useful supplement. Conclusion::The method of AHP and GCD-based multi-index comprehensive evaluation is simple and comprehensive to evaluate the diversity of different PR germplasm resources, and could provide a reference for the development and utilization of resources and the screening of high-quality provenances.
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Objective:To propose a new method for detecting and evaluating traditional Chinese medicine （TCM） by artificial intelligence and machine vision technology. Method:Taking Fritillariae Cirrhosae Bulbus， Crataegi Fructus and Pinelliae Rhizoma as the research objects， big data of pictures was collected by machine vision and the image database was established. Through the intelligent analysis of the external characteristics of TCM， the deep convolutional neural network model was established to realize the functions of location detection and variety identification by means of deep learning， so as to significantly improve the accuracy of rapid identification of TCM. Result:The classification accuracy of 11 kinds of Chinese herbal pieces （raw， fried， parched and charred products of Crataegi Fructus， Pinelliae Rhizoma， Pinelliae Rhizoma Praeparatum Cum Zingibere et Alumine， Pinelliae Rhizoma Praeparatum， Pinelliae Rhizoma Praeparatum Cum Alumine and three products of Fritillariae Cirrhosae Bulbus） could be more than 99%， and the average recognition accuracy of specific categories could reach more than 97%. Conclusion:The intelligent identification technology of TCM decoction pieces realized by deep learning algorithms has the advantages of simplicity， rapidity， high precision and quantifiable detection， which can provide technical support for the quality detection and evaluation of TCM， and enrich the research ideas of quality evaluation of TCM.
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Objective:By the method of network pharmacology， the mechanism of Pinelliae Rhizoma and Citri Exocarpium Rubrum in the treatment of metabolic syndrome was explored. Method:Effective components of Pinelliae Rhizoma and Citri Exocarpium Rubrum were retrieved by TCMSP database，and then selected by ADME parameters. TCMID，BATMAN-TCM，SymMap，TCM-MESH database were used to supplement effective components of TCMSP. TCMSP target prediction model was used to predict potential targets of drugs. DRUGBANK，DisGeNET，CTD，GeneCards，OMIM，PharmGkb，KEGG，DiGSeE databases were retrieved to obtain the targets of metabolic syndrome，and the chips were downloaded and analyzed through GEO database No.GSE98895 to screen out the differential genes of normal people and patients with metabolic syndrome，and supplement the target databases of metabolic syndrome. The intersections of Pinelliae Rhizoma-Citri Exocarpium Rubrum and metabolic syndrome disease targets were obtained by Rstudio 3.6.2. The above intersection targets were imported into the Metascape database for module analysis and overall GO（Biological Process），KEGG and Reactome pathway analysis. The core targets were selected from the intersection targets by using the cytohubba plug-in，the core genes were input into the database of BioGPS，Genecards to analyze the tissue distribution and subcellular distribution，and the core targets were assigned by using the database of DisGeNET. Result:A total of 34 active components and 120 targets of Pinelliae Rhizoma and Citri Exocarpium Rubrum were screened out，and 115 targets were obtained after the intersection of Rstudio 3.6.2. The results of Metascape module analysis and whole analysis were mostly the same，mainly involving the biological processes， such as ligand receptor interaction，calcium ion，cGMP-PKG，cholinergic synapse，thyroid hormone，insulin. The cytohubba plug-in was used to screen out 17 targets，involving 17 key genes， such as VEGFA and NOS3. The tissue and subcellular distribution of the core targets mainly included lymphoblasts，CD33+_myeloid cells，amygdala，pineal and cytoplasmic matrix，mitochondria. The main proteins were signal molecules，kinases and nucleic acids. Conclusion:Pinelliae Rhizoma and Citri Exocarpium Rubrum could treat metabolic syndrome through complex biological processes and pathways，such as blood circulation，ligand receptor interaction of nerve activity，cGMP-PKG，interleukin-related action，calcium ion. This indicates that traditional Chinese medicine（TCM） treated diseases through multi-component，multi-target，multi biological processes，multi-channel and other ways（which is also proved by the distribution of core genes in the tissue，subcellular and protein ascription information），indicating the superiority of the holism concept of TCM. Erchetang and its similar prescriptions are suitable for treating metabolic syndrome，which also indicates that the principle of "treating different diseases with the same therapy" of TCM is not only reflected at the theoretical level； and network pharmacology needs to be further proved in the combination with experimental verification.
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Banxia Xiexin Tang is one of the classic prescriptions for treating digestive system diseases.In this paper，using the method of bibliometrics，information of Banxia Xiexintang in ancient Chinese medical literatures were collected and screened out 399 effective data from 238 kinds of ancient books．Based on the statistics and analysis of the history，drug composition，main disease and syndrome，principle of prescription，dosage，processing，preparation，decocting and taking methods of Banxia Xiexin Tang，it is found that Banxia Xiexin Tang originated from Treatise on Febrile and Miscellaneous Disease written by ZHANG Zhong-jing，a famous physician in the Eastern Han dynasty，it is composed of seven herbs，namely，Pinelliae Rhizoma，Scutellariae Radix，Zingiberis Rhizoma，Ginseng Radix et Rhizoma，Glycyrrhizae Radix et Rhizoma Praeparata cum Melle，Coptidis Rhizoma，and Jujubae Fructus. It was mainly used to treat pizheng of mixed cold and heat. Most of the Banxia Xiexin Tang recorded in later generations follow the prescription composition and indications in Treatise on Febrile and Miscellaneous Disease and its clinical application has been extended and expanded，among the 352 literatures with the main diseases and syndromes recorded，the most common （341） were pizheng，accounting for about 96.88%，in addition，it is also used sporadically for shuzheng，malaria，nausea，damp-wen，jaundice，etc. Among the 122 documents with drug dosage records，nearly half of them are identical with the original records in Treatise on Febrile and Miscellaneous Disease. Among the 112 literatures with pharmaceutical processing，licorice （86） was the most，most of them were roasted （80），followed by pinellia ternata （79），and most of them were "decoction washing and sliding" （67）. Among the 111 documents recorded in the decoction method，most of them inherited Treatise on Febrile and Miscellaneous Disease by "removing the dross and again cooking" （68），there are also "Water Decoction" （32） and "Ginseng Radix et Rhizoma and Jujubae Fructus Decoction" （11）. Among the 108 documents with the method of taking medicine，nearly half of them inherited Treatise on Febrile and Miscellaneous Disease，which is "a liter of warm taking ，three times daily". Based on the ancient Chinese medical literatures，Banxia Xiexintang was systematically analyzed in order to provide more accurate ancient literature reference for the clinical application and development of classic prescriptions.
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Objective: To study the physiological and biochemical characteristics of four dominant microorganisms and the yellow pigment content of Pinelliae Rhizoma Fermentata (PRF), and provide basis for exploring the mechanism of PRF processing. Methods: The optimum growth temperature and pH value of the four dominant microorganisms Bacillus subtilis, Paecilomyces variotii, Byssochlamys spectabilis, and Aspergillus niger were studied. The ability of producing acidase, amylase, protease, and yellow pigment were determined. The yellow pigment content of each sample at different fermentation time points in process of PRF was determined. Results: The most suitable growth temperatures for B. subtilis, P. variotii, B. spectabilis, and A. niger were 35 ℃, 29 ℃, 29-31 ℃, and 39 ℃; And the optimum pH were 7.0, 7.0, 7.5, and 7.0, respectively. Four kinds of microorganisms had the ability to produce amylase and protease. P. variotii and B. spectabilis had the ability to produce yellow pigment. The content of yellow pigment were 69.875, 69.875, 71.750, 119.500, and 137.875 μg/g in the samples at different time points. Conclusion: Four kinds of dominant microorganisms may play an important role in fermentation process of PRF.
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To reveal the medication patterns of Chinese patent medicine in the treatment of cough based on the analysis of the patent status of Chinese materia medica (CMM) in the field of cough control in nearly 20 years. Patents of Chinese patent medicine in the treatment of cough were systematically searched in SIPO Platform and CNKI, association analysis and network analysis of high-frequency medicines were used to reveal the medication patterns of Chinese patent medicine in the treatment of cough with software Clementine 12.0. The results showed that Chinese medicine with highest frequency in 316 formulas was Glycyrrhizae Radix et Rhizoma, the other Chinese herbs with higher frequency were Platycodonis Radix, Ephedrae Herba, Citri Reticulatae Pericarpium, Pinelliae Rhizoma, etc. The main categories were the medicines used to treat cough and asthma, as well as to reduce phlegm. The commonly used herbal pairs were Glycyrrhizae Radix et Rhizoma-Platycodonis Radix, Glycyrrhizae Radix et Rhizoma-Citri Reticulatae Pericarpium, Pinelliae Rhizoma-Glycyrrhizae Radix et Rhizoma, Ephedrae Herba-Glycyrrhizae Radix et Rhizoma, etc. It was concluded that the medication patterns of Chinese patent medicine in the treatment of cough can be revealed based on the analysis of the herbal frequency, herbal pairs, association rules, and the network of high-frequency medicines and the theory of traditional Chinese medicine, so as to provide reference evidences for clinical medication.
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Objective:Screen out the antitumor constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma base on system pharmacology with chemical constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma as study objects, in order to provide the theoretical basis for the development of antitumor and nontoxic activities of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma. Method:The small molecule ligand library of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma was built based on Traditional Chinese Medicine Systems Pharmacology(TCMSP), energy of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma was matched with the key protein targets of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signal pathway by molecular docking (SYBYL2.1, Tripos), the Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma-targets network model was established based on Cytoscape 3.5.1, and the physicochemical properties of the antitumor activity in Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma were predicted by using SwissADME and admetSAR. Result:There were 25 small molecule constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma. Through the energy match, key antitumor constituents of Pinelliae Rhizoma were gondoic acid, 10,13-eicosadienoic, baicalin, 12,13-epoxy-9-hydroxynonadeca-7,10-dienoic acid. Key antitumor constituents of Aconiti Lateralis Radix Praeparata were deltoin, sitosterol, neokadsuranic acid B, 11,14-eicosadienoic acid. Phosphatidylinositol 3-kinase (PI3Kα), phosphatase and tensin homolog deleted on chromosome ten (PTEN), phosphoinositide dependent protein kinase 1 (PDK1) were key antitumor targets of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma. There were 8 key antitumor constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma, which had a low CYP450 inhibition and basically followed the Lipinski rule. Conclusion:Antitumor nontoxic constituents of Aconiti Lateralis Radix Praeparata-Pinelliae Rhizoma and key targets are screened out from the molecular level, which provides the new ideas for the effective use of nontoxic traditional Chinese medicine(TCM) and breaks the restrictions in using nontoxic TCM.
ABSTRACT
Objective: To identify the chemical constituents of Pinelliae Rhizoma(BX)by ultrahigh-pressure liquid chromatography coupled with electrospray time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Method: Chromatograpic separation was performed on a shim-pack xR-ODS Ⅲ column (2.1 mm×75 mm,1.6 μm) using a gradient elution program with mixtures of acetonitrile and 0.1% formic acid-water as mobile phases at a flow rate of 0.25 mL·min-1. UPLC-Q-TOF-MS/MS was developed for rapid and high-throughput screening of the preliminary chemical profile of BX in positive ion modes. Result: Ninety chemical components were found in BX,according to the accurate Charge-mass Ratio and the MS/MS data,retention time of reference standard,references or databases,the fragmentation regularities of mass spectra. Eighty were identified preliminarily,including 7 alkaloids,8 poly-alcohols,12 fatty glycerides,5 flavonoids,12 lysophosphatidylcholines (LPCs),10 alcohol amines,11 amino acids,11 amides and 4 other type. Conclusion: LPCs were first found in BX. Because BX has certain toxicity,it is found that BX contains LPCs by analyzing the chemical constituents of BX,which can cause inflammatory reaction and neuronal myelin sheath loss and degeneration. Therefore,the analysis of the chemical composition of BX can explain the causes of the toxicity and provide a foundation for the basic research and quality control of the potency of BX.
ABSTRACT
Objective: To study the suspension culture of tuber and its alkaloid content based on the stimulation of salicylic acid. Method: The tubers of Pinelliae Rhizoma in suspension tube were treated with different concentrations of exogenous salicylic acid to analyze the growth status. The content of alkaloids in tuber was detected by HPLC. Test conditions:chromatographic column for Agilent Eclipse plus C18 column (4.6 mm×250 mm,5 μm),mobile phase of acetonitrile water(4:96),the column temperature was maintained at 35℃,detection wavelength for inosine 250 nm,guanosine 260 nm,volume flow rate 1.0 mL·min-1. Result: The results showed that the exogenous salicylic acid had a certain effect on the growth of suspension tuber of Pinelliae Rhizoma. When the salicylic acid concentration was 150 μmol·L-1,the culture lasted for 25 days and the fresh weight reached the maximum value of 7.483 8 g. It also accumulates a certain amount of alkaloids. The linear range of guanosine was 0.03-0.45 μg (R2=0.999 6). After 10-days cultivatation in the salicylic acid concentration of 50 μmol·L-1,guanosine content of Pinelliae Rhizoma tubers reached a maximum of 1.353 3 mg·g-1. The linear range of inosine 0.003-0.045 μg (R2=0.999 5). When the salicylic acid concentration was 200 μmol·L-1,cultured for 30 days,the content of inosine in Pinelliae Rhizoma tubers reached the maximum value of 0.149 8 mg·g-1. Conclusion: The results of this experiment provide a reference for the study of tissue culture and rapid propagation of Pinelliae Rhizoma tubers and regulation of alkaloids,which are of great significance for the development of Pinelliae Rhizoma industry.