Inokosterone Is A Potential Drug Target of Estrogen Receptor 1 in Rheumatoid Arthritis Patients: Analysis from Active Ingredient of Cyathula Officinalis / 中国结合医学杂志

OBJECTIVE@#To elucidate the active compounds and the molecular mechanism of Cyathula Officinalis as a drug treatment for rheumatoid arthritis (RA).@*METHODS@#The target genes of active ingredients from Cyathula Officinalis were obtained from bioinformatics analysis tool for the molecular mechanism of traditional Chinese medicine. The protein-protein interaction between the target genes were analyzed using STRING and Genemania. The transcriptome of RA patients compared to healthy people (GSE121894) were analyzed using R program package Limma. The relative expression of the target genes was obtained from the RNA-seq datasets. The molecular docking analyses were processed based on the molecular model of estrogen receptor 1 (ESR1) binding with estradiol (PDB ID:1A52). The binding details were analyzed by SYBYL.@*RESULTS@#Inokosterone, ecdysterone, and cyaterone were the 3 active ingredients from Cyathula Officinalis that bind to target genes. Of all the significantly changed genes from RA patients, ESR1, ADORA1, and ANXA1 were significantly increased in mRNA samples of RA patients.@*CONCLUSION@#ESR1, the transcription factor that binds inokosterone in the molecular binding analysis, is the target protein of Cyathula Officinalis.

Morphological Identification of Cyathula officinalis Kuan. and Cyathula capitata(Wall.) Moq / 中国药学杂志

OBJECTIVE: To identify morphological characteristics of Cyathula officinalis Kuan. and Cyathula capitata(Wall.) Moq.. METHODS: Based on tissue morphology features, C. officinalis Kuan. and Cyathula capitata (Wall.) Moq. were differentiated by using tissue sectioning, optical microscope, scanning electron microscopy and transmission electron microscopy technology. RESULTS: C. officinalis exhibited features, such as main roots, stems composed of four vascular bundles, relative rich trichomes, less green color, however, for C. capitata, hairy roots, stems composed of two vascular bundles and less trichomes were observed. Compared with low altitude condition, high-altitude condition may induce darker green color and formation of yellow spots and osmiophilic granules of leaves for C. capitata. CONCLUSION: It provides a scientific approach to differentiate these two closely related medicinal plants.

Fast Identification of Cyathula officinalis and Its Adulterant and Doped Adulterant Based on Properties and Foam Test / 中国实验方剂学杂志

Objective: To rapidly identify Cyathula officinalis and its adulterant C. capitata and C. officinalis*C. capitata and doped adulterant. Method: Properties combined with foam test method were used for identifying C.officinalis, its adulterant and doped adulterant. In the aspect of properties,6 aspects including shape,size,texture,color,smell and taste,were observed, smelled and tasted. In the aspect of foam test,the volume of foam produced was used as the determination index to investigate the sample amount,water amount, shaking time,particle size,water temperature,repeatability,adulteration ratio and its stability. Result: In the aspect of properties,C. officinalis and its adulterant showed obvious difference in the shape,size,color,texture,smell and taste,especially the red color and bitter taste of its adulterant. In the aspect of foam test,the optimum parameters were as follows:sample particle (screened with 3 sieves) 0.3 g,a test tube with plug and scale,water 10 mL and airtight,forced shaking up and down for 1 min,settling for 5 min. Such method can be used to identify C. officinalis, its adulterant and doped adulterant. The volume of foams produced by C. officinalis and its adulterant and different ratio of doped adulterant showed no change within 5-30 min,slightly decreases after 9 h; the higher adulteration ratio; the larger volume of foam and better stability. The 8 batches of C. officinalis and 8 batch of adulterants proved that the volume of the foams produced was all less than 2 mL in the C. officinalis,more than 13 mL in the adulterant is,and more than 5 mL in 5% doped adulterant, showing statistical difference. From the properties combined with foam test,5 specific identification elements were obtained for identifying C. officinalis, its adulterant and doped adulterant. Conclusion: Through the 5 specific identification elements,the properties combined with foam test can be used to distinguish the C. officinalis from its adulterant C. officinalis and C. officinalis*C. capitata and doped adulterant,characterized by accuracy,simpleness,short time,low cost and feasibility. It can provide a new method and reference for identifying C.officinalis from its adulterant and doped adulterant.

Allele-specific PCR to identification of Achyranthis Bidentatae Radix and Cyathulae Radix formula granules / 中国中药杂志

To establish a robust and accuracy molecular method to identify Achyranthis Bidentatae Radix and Cyathulae Radix formula granules. ITS sequences of Achyranthes bidentata and Cyathula officinalis were aligned, specific SNPs (single nucleotide polymorphisms) were excavated, specific primers were designed and allele-specific PCR method was established. The genomic DNA was successfully extracted from the herbal medicine and its formula granules by using an improved CTAB (cetyltrimethyl ammonium bromide) method and then performed PCR with the designed primers. The 187 bp specific band could be amplified only in the presentation of C. officinalis and its granules when use of C. officinalis specific primers, whereas the 162 bp band could be amplified only in the presentation of A. bidentata and its granules when use of A. bidentata specific primers. This method was also successfully applied in the identification of commercial formula granules.

Application of the Electronic Tongue in the Identification of Cyathula officinalis / 中国药房

OBJECTIVE:To establish the method for "taste" identification of Cyathula officinalis.METHODS:The "taste" of C.officinalis was detected by using electronic tongue:sampling temperature of 25 ℃,sampling time of 120 s,sampling period of 1 s,rolling speed of 1 r/s,cleaning solution of purified water.The response value of electronic tongue sensor at 120 s were used as "taste" analysis data and obtained "taste" related data was analyzed through principal component (PC) analysis and discriminant factor (DF) analysis.C.officinalis and its adulterants,C.officinalis from different regions,different ages and different batches were distinguished.RESULTS:There was significant difference in DF between one batch of C.officinalis and 4 batches of adulterants.The samples from Sichuan Baoxing,Jinkouhe,Tianquan could be clustered together.There was significant difference in DF between annual or biennial samples and triennial or quadrennial samples.There was also significant difference in DF among different batches.CONCLUSIONS:The "taste" is different significantly between C.officinalis and its adulterants,among different regions,different ages and different batches of samples.Electronic tongue can quickly distinguish "taste" of above medicinal materials.

Feature Extraction and Image Recognition of Achyranthes bidentata and Cyathula officinalis / 中国药房

OBJECTIVE:To extract the feature of Achyranthes bidentata and Cyathula officinalis,and to establish image recog-nition method. METHODS:The microscopic image stitching of A. bidentata and C. officinalis was implemented by MATLAB. The color,invariant moment,stripes and the features of vascular bundle in cross section were extracted. The data was organized into da-ta matrix,and then data matrix was standardized by Zscore function;principal components were analyzed through Princomp func-tion. BP nerve network recognition mode was adopted. RESULTS:The microstructures in the micro images of the samples were kept integrated. The measured data of 27 characteristics were acquired in each group of sample. Through principal component analy-sis,the parameters of 11 main components were selected to establish BP never network. The average recognition rate of BP nerve network was 100% between 2 medicinal material relatives (n=50). CONCLUSIONS:The method can be used for micro-image auto Stitching of Chinese medicinal materials and image recognition of A. bidentata and C. officinalis.

Predictive distribution and planting GAP of Cyathula officinalis in China based on 3S technology and MaxEnt modelling / 中国中药杂志

Research on predictive distribution and planting GAP of Cyathula officinalis in China is helpful to provide scientific basis for its protection and planting popularization. According to the data in 63 distribution sites and 49 ecological variables, using MaxEnt ecological niche model and 3S technology, we performed a quantitative analysis of suitable distribution and planting GAP of C. officinalis in China. Our results show that: ① the area of suitable distribution of C. officinalis is about 634 385.80 km² in total, and mainly in Northeastern and Southeastern Sichuan, Northern and Southeastern Yunnan, Western and Southwestern Guizhou, Southwestern and Northeastern Chongqing, Southwestern Shaanxi, Southeastern Gansu, Western Guangxi, Southeastern Tibet. ② The main ecological factors determining the potential distribution are precipitation, altitude, minimum temperature of coldest month, soil type, monthly mean temperature. ③ The planting GAP region are mainly in Guangyuan, Mianyang, Ya'an, Leshan, Liangshan, Panzhihua of Sichuan province, Hanzhong of Shaanxi province, Dali, Nujiang, Chuxiong, Baoshan, Qujing, Wenshan of Yunnan province, southwestern autonomous prefecture in Guizhou province. The results are of great significance for realizing the growth environment, predicting the potential distribution and promoting planting popularization for C. officinalis.

Triterpenoid saponins from the roots of Cyathula officinalis and their inhibitory effects on nitric oxide production / 中国天然药物

The present study was designed to investigate the chemical constituents of the roots of Cyathula officinalis. Compounds were isolated by silica gel, Sephadex LH-20, ODS column chromatography, and preparative HPLC. Their structures were determined on the basis of 1D and 2D NMR techniques, mass spectrometry, and chemical methods. One new oleanane-type triterpenoid saponin, 28-O-[α-L-rhamnopyranosyl-(1→3)-β-D-glucuronopyranosyl-(1→3)-β-D-glucopyranosyl] hederagenin (1), was isolated from the roots of Cyathula officinalis. The anti-inflammatory activities of the isolates were evaluated for their inhibitory effects against LPS-induced nitric oxide (NO) production in RAW 264.7 macrophages cells. Compounds 2, 4, and 6 exhibited moderate anti-inflammatory activities.

HPLC fingerprint of crude Cyathula officinalis and its processed products stir-fried with wine and salt and variation of their index components / 中草药

Objective: To reveal the variation regularity of chemical components in crude Cyathula officinalis and its processed products (stir-fried with yellow rice wine and salt), and promote the quality control of the herb. Methods: Agilent Zorbax Eclipse XDB-C18 column (250 mm×4.6 mm, 5 μm) was used with acetonitrile and 0.1% phosphoric acid solution in gradient elution mode. The detective wavelength was 266 nm and the flow rate was 0.5 mL/min. The fingerprints for 24 herbal samples were set up and 25 peaks were recorded with different retention time and peak areas. Three peaks were successfully identified as puerarin, cyasterone, and daidzein by comparing with the retention time of reference substances. The vectorial angle method was used to evaluate the similarity between fingerprints. The cluster analysis and principal component analysis were applied to studying the HPLC fingerprint and chemical pattern recognition. Results: The two processing techniques, stir-frying with wine and salt, both had significant effect on herbal chemical profiles. The contents of three known active components, puerarin, cyasterone, and daidzein, were not observed except a little increasing of the content of puerarin after the crude herbs were processed with salt. Conclusion: More precise active components of the processed products of C. officinalis need to future study to improve the current quality standard of C. officinalis in Chinese Pharmacopoeia. The method provided by this study is a powerful tool to identify and quality control between crude and processed C. officinalis because of its quantificational and visual evaluation system.

Study on suitable distribution areas of Cyathula officinalis in Sichuan province based on remote sensing and GIS / 中草药

Objective: To evaluate the suitability of Cyathula officinalis resources grown in Sichuan province, and provide a reference of the monitoring for traditional Chinese medicine resource. Methods: Taking the planting of C. officinalis in Sichuan province as an example, The topography, climate, soil, and vegetation of C. officinalis were researched using remote sensing and GIS techniques, binding environment factors in growing and adopting the analytic hierarchy process (AHP) and spatial analysis method. According to the different classification of index factors, the classification map of the suitable area for C. officinalis was obtained and the field measurements were combined to verify. Results: The C. officinalis resources in Sichuan province were mainly distributed in about 19 cities or autonomous prefectures such as Leshan City, Ya'an City, Yi Autonomous Prefecture of Liangshan, and Yibin City with suitability distribution areas of 3 633.52 km2 approximately and accounting for 0.76% of the total area. According to the field investigation, related documents, and information records, the suitability distribution based on RS and GIS correspond with the actual distribution areas of C. officinalis resources. Conclusion: It is a feasible method to divide the suitability distribution area of C. officinalis Kuan by using RS and GIS as quickly and accurately as possible. It can also provide a support on scientific and date basis to a complete investigation on suitable distribution areas, protection, rational development and utilization of C. officinalis and other medical materials.

Cloning, subcellular lacalization and expression analysis of chloroplast-targeted Obg gene in Cyathula officinalis / 中国中药杂志

According to ObgC gene sequences from Cyathula officinalis genomic data, the specific primers were designed, and a full-length CoObgC cDNA (2 226 bp) was obtained by polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) methord. Sequence alignment showed that CoObgC gene contained a 1 818 bp open reading frame (ORF) encoding 605 amino acids. Sequence analysis predicted that molecular weight of CoObgC protein was about 66.39 kDa, the academic isoelectric point was 5.35, and the protein was stable protein. Then multiple sequence alignment was applied to construct phylogenetic tree. The real-time fluorescence quantification PCR (RT-qPCR) demonstrated that a high expression level in leaf, followed by root and flower, the low transcription was in stem. The recombinant vector pCABIA2300-CoObgC was constructed and introduced into tobacco epidermal cells by agrobacterium-mediated transformation, green fluorescence was tested and targeted to chloroplast under a laser scanning confocal microscope. These findings will be helpful to lay a foundation for studying the structure and function of CoObgC gene, and elucidating C. officinalis molecular biology experiment.

Effect of moisture content on vigor of Cyathula officinalis seeds and its anti-aging mechanism analysis / 中国中药杂志

Effects of nine different moisture contents on vigor of Cyathula officinalis seeds and its anti-aging mechanism were studied by artificial accelerated aging through high temperature and wet. The research results showedthat seed vigor were generally decreased after artificial aging; in general, seed vigor and its anti-aging ability are relatively stronger within the scope of 6.55%-4.78% moisture content, the increase range of seed conductivity, peroxidase activity, malondialdehyde content,and reduce amplitude of activityof dehydrogenase , superoxide dismutaseare alllower as well. And when the moisture content reduced to 5.77%, all of the germination tests index of the non-aged seeds are the highest, and the activity of peroxidase the lowest,conductivity of leaching solution relatively low, activity of dehydrogenase and superoxide dismutase the highest,and catalase activityrelatively high.Therefore, in the low temperature germplasm preservation of C. officinalis seeds, the seed moisture content should be controlled close to the range of (5.70±1)% to keep higher vigor and anti-aging ability.

Grey relational analysis of the relationship between variety and quality of radix cyathulae / 中国药学杂志

OBJECTIVE: To screen excellent provenance of Radix Cyathulae and ensure the quality of Radix Cyathulae.

EXPERIMENTAL STUDY ON ANTIFERTILITY EFFECT OF CYATHULA OFFICINALIS KUAN / 西安交通大学学报(医学版)

The antifertility and antinidation effects of three extracts (benzol, ethyl acetate and alcohol) of Cyathula Oficinalis Kuan on mice were observed. The results indicate that the benzol extract had significant antifertility action. Its effective rate of antifertility was 100%, when 250 mg/kg was given orally. As compared with control, when 250 mg/kg ethyl acetate extract was given, the rate of antifertility was 63.7%. If 500 mg/kg ethyl acetate extract was given the effective rate of antifertility increased over 90%. However, when 250～500 mg/kg alcohol extract was given, the rate of antifertility was only 40～54%. The study of antinidation showed that the benzol extract (500 mg/kg )could effectively prevent implatation. The rate of antinidation was 100%, and yet the ethyl acetate and alcohol extracts were 66.7% and 40% repectively. The results suggest that the three extracts of Cyathula Officinalis Kuan all had antifertility effect, but the action of benzol extract was more effective than other two extracts.