Herbal Textual Research on Chrysanthemi Flos in Famous Classical Formulas / 中国实验方剂学杂志

To conduct textual research on the records of Chrysanthemi Flos in the ancient literature from the aspects of name， origin， species， scientific name， origin， harvesting， processing， quality and so on， and the modern literature was sorted out to clarify the relationship between the ancient and modern times， so as to provide a reference for the development of famous classical formulas containing Chrysanthemi Flos. Chrysanthemi Flos is an ornamental， edible and medicinal plant in China， it has many aliases， but it has been recorded in this materia medica under the name of Juhua， Ganju and Ganjuhua. Before the Tang dynasty， medicinal Chrysanthemi Flos mainly collected wild products， including yellow flowers and white flowers， of which the mainstream of yellow flowers was originally Dendranthema lavandulifolium and D. potentilloides， the mainstream of white flowers is D. vestitum and D. chanetii. The cultivation of medicinal Chrysanthemi Flos began in the Northern Song dynasty， and wild D. lavandulifolium， D. potentilloides， D. vestitum and D. chanetii were selected through long-term interspecies and intraspecies crossbreeding， which gradually formed the current cultivar D. morifolium. After chrysanthemums were introduced abroad， foreign scholars began to name chrysanthemums with Linnaeus's plant classification system. In 1792， Mathier named chrysanthemums as Chrysanthemum morifolium and continued to this day， and all the editions of Chinese Pharmacopoeia adopted this scientific name. In the Song dynasty， many local varieties such as Nanyangju， Dengzhouhuang and Dengzhoubai appeared. By the time of the Republic of China， five famous authentic varieties， namely Huaiju， Boju， Chuju， Gongju and Hangju， had been cultivated for medicinal purposes. Boju has been the best medicinal variety since the late Qing dynasty. Hangbaiju has been famous for its tea use， especially the best quality of Huju. Chuju has its own unique characteristics， and it is of good quality both for medicine and tea. Gongju has always been a good tea chrysanthemum. Chrysanthemums are traditionally harvested in September of the lunar calendar， but some of the new varieties cultivated nowadays are harvested earlier. The embryo chrysanthemum in Hangbaiju is a commodity type that collects unopened buds in advance. In ancient times， chrysanthemums were mainly dried in the shade， in modern times， drying methods include drying in the shade， drying in the oven and drying in the sun after steaming. At present， hot air drying is mostly used. In terms of processing， Chrysanthemi Flos was used raw products in ancient times， in modern times， it is still widely used， sometimes stir fried（including stir-fried charcoal）. Due to different varieties， producing areas and processing methods， there are certain differences in the proportion of ingredients contained in chrysanthemum. Therefore， it is suggested that chrysanthemums with different varieties， origins and processing methods should be selected according to clinical indications in the development of famous classical formula preparations containing Chrysanthemi Flos.

Geju Hugan Tablets Ameliorate Alcohol-induced Liver Injury in Mice by Regulating NF-κB and Bcl-2/Bax Signaling Pathways / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of Geju Hugan tablets on the liver of mice with alcohol-induced liver injury， and explore the underlying mechanism based on nuclear factor-κB p65 （NF-κB p65） and B-cell lymphoma-2 （Bcl-2）/Bcl-2-associated X protein （Bax） signaling pathways. MethodAccording to the body weight， 60 SPF-grade male ICR mice were randomized into normal， model， Compound Yiganling tablets （0.16 g·kg-1）， and low-， medium-， and high-dose （0.2， 0.4， 0.8 g·kg-1， respectively） Geju Hugan tablets groups. The drugs were administrated at the corresponding doses by gavage， and the normal and model groups with equal volume of pure water once a day for 28 consecutive days. On day 29， the mice in other groups except the normal group were administrated with liquor （53% Vol） by gavage twice a day at the doses of 20， 10 mL·kg-1 and with the interval of 6 h. Samples were harvested on day 30. The histopathological changes in the liver were observed by hematoxylin-eosin （HE） staining， and the ultrastructural changes in hepatocytes were observed by transmission electron microscopy. The enzyme-linked immunosorbent assay was employed to measure the levels of malonaldehyde （MDA）， reduced glutathione （GSH）， and triglycerides （TG） in the liver tissue and the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in the serum. Western blotting was employed to determine the protein levels of NF-κB p65， phosphorylated p-inhibitor kappa B alpha （p-IκBα）， Bcl-2， and Bax in the liver tissue. ResultCompared with the normal group， the model group showed increases in the ALT， AST， MDA， and TG levels， a decrease in the GSH level， and increases in the liver injury scores evaluated based on the HE， oil red O， and transmission electron microscopy （P<0.01）. Moreover， the model group showed up-regulated expression of NF-κB， p-IκBα， and Bax （P<0.05， P<0.01） and down-regulated expression of Bcl-2 （P<0.05） in the liver tissue. Compared with the model group， Geju Hugan tablets of all the doses lowered the ALT， AST， MDA， and TG levels and elevated the GSH level （P<0.01）. The liver injury scores assessed based on HE staining and transmission electron microscopy in the medium- and high-dose Geju Hugan tablets groups were lower than those in the model group （P<0.01）. Compared with the model group， medium- and high-dose Geju Hugan tablets down-regulated the protein levels of NF-κB， p-IκBα， and Bax （P<0.01） and all doses of Geju Hugan tablets up-regulated the protein level of Bcl-2 （P<0.01）. ConclusionGeju Hugan tablets protect mice from alcohol-induced liver injury by down-regulating NF-κB signaling pathway to alleviate inflammation in the liver tissue and down-regulating the expression of Bax and up-regulating the expression of Bcl-2 to inhibit hepatocyte apoptosis.

Analysis of Chemical Constitutions of Volatile Oil in Opisthopappus taihangensis by GC-MS Combined with Retention Index / 中国实验方剂学杂志

ObjectiveTo study the composition of volatile oil in Opisthopappus taihangensis （Taihangju）， and provide a reference for comprehensive development of this medicine. MethodTaking Chrysanthemum morifolium （Xiaobaiju）， C. morifolium （Xiaohuangju） and C. indicum （Yejuhua） as control， the qualitative and quantitative analysis of volatile oil in Taihangju and three control varieties were completed by gas chromatography-mass spectrometry （GC-MS） combined with retention index method. The GC conditions were as following：programmed temperature （initial temperature at 60 ℃， kept for 2 min； up to 120 ℃ with the heating rate of 5 ℃·min-1， still kept for 2 min； up to 180 ℃ with the heating rate of 2 ℃·min-1， kept for 3 min； and then up to 240 ℃ with the heating rate of 8 ℃·min-1， kept for 5 min； finally up to 280 ℃ with the heating rate of 10 ℃·min-1， kept it for 5 min and finished）， high-purity helium as the carrier gas， the split ratio of 50∶1. MS conditions were as follows：electron impact ion source （EI）， ion source temperature of 230 ℃， electron collision energy of 70 eV and scanning range of m/z 30-445. Principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were used to obtain the characteristic components between Taihangju and the three control varieties. ResultA total of 86， 96， 112 and 109 compounds including 73 common components were identified in Taihangju， Xiaobaiju， Xiaohuangju and Yejuhua， respectively. The contents of volatile components in Taihangju were significantly different from that of the control varieties. In which， the relative contents of α-thujone， eucalyptol and terpinen-4-ol were high in Taihangju， and eucalyptol， camphor and α-terpinyl acetate were the main compositions in the control varieties. In addition， 11 compounds were screened as characteristic components to distinguish Taihangju and the three control varieties by PCA and OPLS-DA， including main differential components of chamazulene and δ-cadinene. ConclusionThe main components of volatile oil in Taihangju includes alcohols， terpenes， ketones and esters with high medicinal value. The accuracy of qualitative analysis of volatile oil is improved by GC-MS combined with retention index method， which provides scientific reference for development and utilization of Taihangju.

Evaluation and optimization of content determination method of Chrysanthemi Flos in Chinese Pharmacopoeia / 中国中药杂志

This study discovered that the resolution of 3,5-O-dicaffeoylquinic acid(isochlorogenic acid A) in the content determination method of Chrysanthemi Flos in Chinese Pharmacopoeia(ChP)(2020 edition) was poor, which affected accurate quantification. We tested the method in ChP with chromatographic columns of seven brands to clarify the problems in the existing method, optimized the chromatographic conditions by adjusting the mobile phase composition and elution ratio and replacing the chromatographic column packing, and carried out the reproducibility assay for the new method. The two methods were compared for the content determination results of Chrysanthemi Flos prepared from six different varieties. As evaluated by the resolution based on different chromatographic columns of seven brands, the existing method failed to separate isochlorogenic acid A and isochlorogenic acid D well. The peaks of the two components were not completely separated on three chromatographic columns, and isochlorogenic acid A and isochlorogenic acid D generated a co-effluent peak in the other four columns. Isochlorogenic acid A and isochlorogenic acid D could be completely separated under the optimized chromatographic conditions. The difference in the peak areas of isochlorogenic acid A+isochlorogenic acid D obtained by the optimized method and the method in ChP was not significant, with deviation less than 3.0%, which further proved that the result measured by the method in ChP was the co-effluent of isochlorogenic acid A and isochlorogenic acid D. The optimized method can ensure the accurate quantification of isochlorogenic acid A. The existing content determination method of Chrysanthemi Flos has the problem of poor resolution. It is recommended to revise the chromatographic conditions for the content determination method of Chrysanthemi Flos to improve the resolution of isochlorogenic acid A and ensure its accurate quantification.

Changes in chemical compositions of Chrysanthemi Flos after frying and protective effects on CCl_4-induced acute liver injury in mice / 中国中药杂志

To reveal the processing mechanism of Chrysanthemi Flos from the changes of chemical compositions after frying and its effect on the efficacy of liver protection. Ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS) and ultra high performance liquid chromatography(HPLC) were used for the qualitative and quantitative researches of chemical compositions before and after Chrysanthemi Flos frying. Progenesis QI and SPSS software were used for principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), variable importance projection(VIP) analysis and t-test to identify the compositions with significant changes. Pharmacodynamics experiment was used to investigate the protective effect of crude and fried Chrysanthemi Flos on CCl_4-induced acute liver injury in mice. According to mass spectrometry data, there were 28 chemical compositions in crude and fried Chrysanthemi Flos, mainly including flavonoids and organic acids. 13 compositions such as luteolin, apigenin and luteolin glycoside were increased significantly after frying, while 7 compositions such as chlorogenic acid, luteolin-7-O-glucuronide and apigenin-7-O-glucuronide were decreased significantly after frying. Through principal component analysis, crude and fried Chrysanthemi Flos products were divided into two categories, indicating that there were internal differences in quality. The results of liver injury protection experiment in mice showed that the AST, ALT and MDA contents were significantly decreased and SOD level was increased in mice with liver injury in both the high and medium dose groups. Histopathological examination showed that crude and fried Chrysanthemi Flos can protect the liver by reducing inflammatory cell infiltration, reducing steatosis, and repairing damaged liver cells. The results of this study showed that the chemical compositions had obvious changes after frying, and both crude and fried Chrysanthemis Flos had protective effects on CCl_4-induced acute liver injury in mice. In addition, in the range of high, medium and low doses, the liver protection effect of crude and fried Chrysanthemi Flos increased with the increase of dose. The experiment results provided reference for the mechanism of fried Chrysanthemi Flos and clinical selection of processed products.

Herbal Textual Research on Genuine Medicinal Materials “Deng Ju” and Quality Evaluation of Nan Yang Chrysanthemi Flos / 中国实验方剂学杂志

Objective: To study the origin of Nanyang Chrysanthemi Flos materia medica and its historical origin, in order to evaluate the quality of Nanyang Chrysanthemi Flos by HPLC method, and define the advantages of Nanyang Chrysanthemi Flos with the origin of Nanyang. Method: Records of Nanyang Chrysanthemi Flos in the "Chinese Medical Code" and related ancient documents were studied to explain the origin and application. The genetic relationship between Nanyang Chrysanthemi Flos and other pieces was revealed in ancient literatures. Then 8 chemical constituents in 14 batches of Chrysanthemi Flos were determined by HPLC multi-components quantitation,and the comprehensive weighted score analysis was performed based on the results. The HPLC fingerprints were established,and the similarity analysis and clustering analysis were made to comprehensive evaluation the quality of Nanyang Chrysanthemi Flos and define the genetic relationship between Nanyang and other pieces at the chemical composition level. Result: The results of the herbal textual research show that Nanyang Chrysanthemi Flos spread from place to place since the Han dynasty,and impact many medicinal chrysanthemums of later generations. HPLC fingerprints similarity and cluster analysis also indicated the genetic relationship between Nanyang Chrysanthemi Flos and other species at the chemical level. The comprehensive score analysis results show that Nanyang Chrysanthemi Flos is of good quality and very suitable for medicinal purposes. Conclusion: Nanyang has been a quality production area of Chrysanthemi Flos since ancient times to present. Nanyang Chrysanthemi Flos is very suitable for medicinal purposes and worthy of promotion.

Imprinting equivalence research of Lonicerae Japonicae Flos and Lonicerae Flos / 中草药

Objective: To explore and confirm the imprinting equivalence of Lonicerae Japonicae Flos (LJF) and Lonicerae Flos (LF) based on the autonomous function of the supramolecular imprinted template, and provide a new solution for double followers dispute. Methods: Using LJF dregs, LF dregs, and Chrysanthemi Flos (CF) dregs as host molecules, and the water extract of LJF, LF, Shuanghuanglian (LJF), Shuanghuanglian (LF), Yinqiaosan (LJF), and Yinqiaosan (LF) as guest molecules, the selective absorb of three host molecules and six guest molecules was carried out. The change of fingerprint of water extract of guest molecules was determinated by HPLC. Then, the MRT and their difference value was calculated through the total quantum statistical moment method,and t-test was performed on it. Results: When the six guest molecules was absorbed by LJF and LF dregs, LJF and CF dregs, LF and CF dregs, the MRT difference value was conducted by t-test. The results were P1 = 0.94 > 0.05, P2 = 0.02 < 0.05, and P3 = 0.04 < 0.05. We can see that all guest molecules was absorbed by LJF and LF dregs. There was no significant difference in the MRT difference value. But when six guest molecules was absorbed by CF dregs. There was significant difference between CF and LJF and CF dreg in the MRT difference value. Conclusion: The statistical data indicated that similarity was existed in both “imprinted template” of LJF and LF. There was difference with CF. It is identified the exist of imprinting equivalence of LJF and LF, which is compatible with clinical medication.

Construction of high quality Chrysanthemi Flos pieces standard system based on comprehensive analysis of ancient and modern Chinese and foreign pharmacopoeia / 中国中药杂志

To study the evolution of Chinese ancient and modern pharmacopoeia standards and compare the domestic and foreign pharmacopoeias, further understand the international requirements on chrysanthemum quality, and establish a more suitable and modern standard system for high quality Chrysanthemi Flos pieces. Newly Revised Materia Medica, Welfare Pharmacy, Collected Essentials of Species of Materia Medica (Bencao Pinhui Jingyao), Chinese Pharmacopoeia and other herbal remedies in various generations were reviewed to summarize the evolution of domestic standards on Chrysanthemi Flos pieces. Then they were compared with those in European Pharmacopoeia, United States Pharmacopoeia, Japanese Pharmacopoeia and other foreign Pharmacopoeias to establish a modern and international high-quality Chrysanthemi Flos pieces standard system with Chinese medicine characteristics and produce more internationally recognized high-quality Chinese medicine pieces.

Research and application of quality standard for standard decoction of Chrysanthemi Flos / 中国中药杂志

Standard decoction of traditional Chinese medicine (TCM) is prepared by standardized process, and can be used as references to evaluate the quality of dosage forms such as decoction and dispensing granules. In order to determine the quality evaluation method for standard decoction of Chrysanthemi Flos and investigate its application, 10 batches of white chrysanthemum of Hangzhou were collected to prepare the standard decoction of white chrysanthemum of Hangzhou with standardized process parameters. Parameters such as traits, relative density, pH value, extraction ratio, transfer rate and fingerprint were selected as the indexes for quality evaluation. The established quality evaluation method for standard decoction of Chrysanthemi Flos was applied in the detection of two types of commercial Chrysanthemum dispensing granules. The results showed that the standard decoction of Chrysanthemi Flos was a clear yellow-brown aqueous solution; the relative density was 1.007-1.011; the pH value was between 5.37-5.56; the average extraction ratio was 23.6%, ranging from 19.93% to 29.69%; the average transfer ratewas 56.2% in terms of chlorogenic acid, 57.4% in terms of luteoloside and 30.6% in terms of 3,5--dicaffeoylquinic acid. Fingerprint similarity was between 0.864-0.989.The method showed good precision, stability and repeatability in fingerprint analysis, indicating reliable and representative results for standard decoction of Chrysanthemi Flos, and it can be used to evaluate and standardize other dosage forms.

Whole-ingredient identification and primary grade evaluation of Niuhuang Qingwei Pills based on reference drug / 中草药

Objective To identify whole ingredients in Niuhuang Qingwei Pills (NQP) and give primary grade evaluation of NQP. Methods Firstly, microscopic characteristics specified by the statutory standard of NQP were summarized. Then new microscopic identification methods were established for Sennae Folium and Gypsum Fibrosum while new thin layer chromatography (TLC) identification methods were established for Bovis Calculus Artifactus, Borneolum Syntheticum, and Rhei Radix et Rhizoma. In addition, microscopic identification method for Menthae Haplocalycis Herba was improved. In this way, whole-ingredient identification of 17 materials were realized. NQP reference drug was developed and applied as accompanying control in whole-ingredient identification and primary grade evaluation of the samples. Results According to the results of 48 samples from 18 manufactures by primary grade evaluation, 10, 37, and 1 samples were classified as superior, qualified, and unqualified, respectively. Conclusion The proposed methods are accurate, simple, and objective, which offers a more comprehensive approach for quality control of NQP. And the methods provide research strategy and experimental basis for further work on establishment of grade standard.

Identification of Sulfur Fumigated Chrysanthemi Flos with Electronic Nose Technology / 世界科学技术-中医药现代化

This study was aimed to identify Chrysanthemi Flosbefore and after sulfur fumigation based on its different odour by the electronic nose technology.It was expected to explore a new method for the Chrysanthemi Flos identification according to the odour.The electronic nose technology was used in the detection of peak response values of Chrysanthemi Flos on sensor array.The principal component analysis (PCA) and 10 machine learning (ML) ways were used in the analysis of response values and establishment of optimized identification models.The results showed that there was a significant difference in the odour between sulfur fumigated Chrysanthemi Flos and non-sulfur fumigated ones.The identification models were successful with high correct judge rate by PCA and 6 ML ways including BF Tree,J48 and Random Tree.It was concluded that the electronic nose technology can be used for the accurate identification of sulfur fumigated Chrysanthemi Flos and non-sulfur fumigated ones.The electronic nose technology combined with multiple ML methods can be introduced in the quality evaluation of Chrysanthemi Flos.It provided more ideas for the application of electronic nose in data mining for traditional Chinese medicine (TCM) studies.

Effect of extract from Trichosathis Fructus, Carthami Flos, Chuanxiong Rhizoma, and Chrysanthemi Flos on oxidative stress-induced gut immunity in Drosophila melanogaster / 中草药

Objective: To study the effect of the extract from Chinese materia medica (CMM) on oxidative stress-induced gut immunity of Drosophila melanogaster. Methods: D. melanogaster was treated by Paraquat and H2O2 with or without CMM, and the effects of 50 kinds of CMM on the survival rate of D. melanogaster in response to oxidative stress were recorded. And the antioxidant effects of CMM on lifespan and the number of ISC and enteroblast (EB) in intestinal epithelium cells were analyzed. Results: The extracts from Trichosanthes kirilowii (Trichosathis Fructus), Carthamus tinctorius (Carthami Flos), Ligusticum chuanxiong (Chuanxiong Rhizoma), and Chrysanthemum monfolium (Chrysanthemi Flos) could significantly improve the survival rate of Paraquat and H2O2-induced D. melanogaster, increase the lifespan of D. melanogaster, and regulate Paraquat-induced and age-related changes in the number of ISC and EB. Conclusion: The extracts from T. kirilowii, C. tinctorius, L. chuanxiong, and C. monfolium could significantly improve the oxidative stress-induced gut immunity of D. melanogaster.