Intelligent Identification of Fritillariae Cirrhosae Bulbus，Crataegi Fructus and Pinelliae Rhizoma Based on Deep Learning Algorithms / 中国实验方剂学杂志

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.

Study on Improvement of Quality Standards of Shiwei Yipi Granules / 中国药房

OBJECTIVE：To prov ide reference for the improvement of quality standards of Shiwei yipi granules. METHODS ： According to the general rules of 2015 edition of Chinese Pharmacopeia （part Ⅳ），microscopic identification was used to identify Massa Medicata Fermentata and Galli Gigerii Endothelium Corneum ；TLC method was used to qualitatively identify Crataegi Fructus and Semen Raphani ；the content of sinapine thiocyanate in Semen Raphani was determined by HPLC. RESULTS ：The microscopic characteristics were obvious for Massa Medicata Fermentata （palisade cells of testa and stone cells of testa ）and Galli Gigerii Endothelium Corneum （irregular fragments ）. The same fluorescent spots of Crataegi Fructus and Semen Raphani were displayed at the same position of ursolic acid ，sinapine thiocyanate control and Semen Raphani reference substance. The linear range of sinapine thiocyanate was 23.27-9 574.42 ng （r＝1.000 0）. The LOD and LOQ were of 0.50 μ g/mL and 1.68 μ g/mL， respectively. RSDs of precision ，repeatability，intermediate precision and stability tests （24 h）were all less than 1.5％. The average recoveries were 99.40％-100.89％（RSDs were 0.18％-0.49％，n＝3）. The contents of sinapine thiocyanate in 3 batches of Shiwei yipi granules were 0.086 4-0.220 6 mg/g，the average was 0.168 4 mg/g. CONCLUSIONS ：The identification method of Massa Medicata Fermentata ，Galli Gigerii Endothelium Corneum ，Crataegi Fructus and Semen Raphani in Shiwei yipi granules as well as the method for content determination of sinapine thiocyanate in Semen Raphani are established successfully. The content of Semen Raphani in the Shiwei yipi granules is no less than 0.16 mg/g calculated by sinapine thiocyanate （C16H24NO·5 SCN）.

Effect of Different Processed Products of Crataegi Fructus on Dyspepsia by 1H-NMR Metabonomics / 中国实验方剂学杂志

Objective:The effect of processing on promoting digestion of Crataegi Fructus was investigated by the method of serum metabonomics based on 1H-NMR and the analysis of traditional biochemical indexes. Method:The dyspepsia model was induced by high protein and calorie diet. The small intestine propulsive rate in mice and the hormone level in rat gastrointestinal tract were used as pharmacodynamic indexes. After administration of Crataegi Fructus, fried and charred Crataegi Fructus, the changes of serum metabolites were analyzed by 1H-NMR technique combined with multivariate statistical analysis, so as to compare the therapeutic effects of each processed products on rats with dyspepsia model. Result:Each processed products group of Crataegi Fructus could improve the ability of intestinal propulsion and gastric emptying, especially in charred Crataegi Fructus group, caused by dyspepsia. The serum metabonomics proved that there were significant metabolic differences between the model group and the blank group. A total of 13 biomarkers related to dyspepsia, covering 3-hydroxybutyric acid, glycerophosphoryl choline (GPC), N-acetylglycoprotein, O-acetylglycoprotein, trimethylamine oxide (TMAO), alanine, acetic acid, glutamic acid, glutamine, creatine, leucine, lactic acid and glucose, were screened on the grounds of VIP value of S-plot, single factor ANOVA and area under receiver operating characteristic curve (ROC curve) (AUC). The metabolite composition of each administration group was the same except that the callback tendency of O-acetylglycoprotein, glutamine and GPC in charred Crataegi Fructus group was closer to that in the blank group. Conclusion:There are differences in the effect of promoting digestion to eliminate stagnation among processed products of Crataegi Fructus, charred Crataegi Fructus is more effective in improving dyspepsia, which may play a role in regulating gastrointestinal motility and energy metabolism.

Effects of different processed products of Crataegi Fructus on hyperlipidemia rat model by 1H-NMR metabolomics / 中草药

Objective: To investigate the difference of lipid-lowering effect of raw and processed Crataegi Fructus, and to select the optimal antihyperlipidemic product. Methods The hyperlipidemia model of SD rats was induced by high fat diet. After administration of Crataegi Fructus (1.5 g/kg), fried and coked Crataegi Fructus (1.5 g/kg), the changes of serum metabolites were analyzed by 1H-NMR technique combined with multivariate statistical analysis. Results :Different processing products of Crataegi Fructus can significantly reduce the levels of TC, TG, LDL-C in hyperlipidemia rats, among which Crataegi Fructus had best hypolipidemic effects. The serum contents of lactate, pyruvate, lysine, creatine, glycerol, ornithine, TMAO, α-ketoisovaleric acid, O-acetylglycoprotein, N-acetylglycoprotein, leucine and isoleucine were increased in model group, while choline, acetone, threonine, scyllitol, glutamine, glutamic acid, MUFA, PUFA, GPC, and glucose were decreased compared with control group. After administration of different processing products of Crataegi Fructus, endogenous metabolites were recovered significantly. The effect of Crataegi Fructus on hyperlipidemia was superior to those of fried and coked Crataegi Fructus according to the degree and number of the metabolites recovered. Conclusion: There are differences between the raw and processed Crataegi Fructus, and raw Crataegi Fructus showed the best effect on hyperlipidemia, the effect may be achieved through regulating oxidative stress, energy metabolism, amino acid metabolism, and gut microbiota.

HPLC fingerprint of Xintongtai Granules / 中草药

Objective: To establish an HPLC method for fingerprint analysis of Xintongtai Granules (XG) for its quality control. Methods: The chromatographic behaviors were obtained by a Phenomenext Luna C18 column (250 mm × 4.6 mm, 5 μm) with gradient elution using 0.1% formic acid solution-methanol as the mobile phase. The detection wavelength was 280 nm, the volume flow rate was 1.0 mL/min, and the column temperature was 30 ℃. The samples of 10 batches of XG were determined, and the chromatographic fingerprint similarity evaluation system of Chinese medicine (2012 edition) was used to establish the common pattern of XG fingerprints, and the similarity was calculated. Then the common peaks were identified by the reference chromatogram. Results: HPLC fingerprints of XG were established by the determination of 10 batches of samples. The similarity was above 0.95. A total of 26 common peaks were calibrated. Three mutual peaks (No. 21, 22, 26 peaks) were from Salviae Miltiorrhizae Radix et Rhizoma, eight mutual peaks (No. 4—10, 16 peaks) were from Puerariae Lobatae Radix, seven mutual peaks (No. 13, 15, 17—20, 23 peaks) were from Aurantii Fructus, No. 12 peak was from Chuanxiong Rhizoma, No. 14 peaks was from Glycyrrhizae Radix et Rhizoma, No. 1 peak was shared by Notoginseng Radix et Rhizoma and Glycyrrhizae Radix et Rhizoma, No. 2 peaks was shared by Notoginseng Radix et Rhizoma, Aucklandiae Radix, and Crataegi Fructus, No. 3 peaks was shared by Chuanxiong Rhizoma and Aucklandiae Radix, No. 11 peaks was shared by Puerariae Lobatae Radix and Curcumae Radix, the No. 24 peaks was shared by Chuanxiong Rhizoma and Aurantii Fructus, and No. 25 peaks was shared by Curcumae Radix and Aurantii Fructus. The common peaks were all assigned to each medicinal material and identified by reference chromatograms: No. 7, 12, 22, and 26 peaks were puerarin, ferulic acid, salvianolic acid B, and tanshinone IIA. Conclusion: The similarity results of 10 batches of samples indicate that the particle preparation process is stable and feasible, and the established HPLC fingerprint method is stable and reliable. It can be used to measure the stability of XG production process and the controllability of finished product quality.

Survey on pesticide use in Crataegi Fructus and analysis of pesticide residues based on LC-ESI-MS/MS / 中国中药杂志

In order to study the pesticide residues of the medicinal Crataegi Fructus,this study aims to establish an analysis method for pesticide residues( mainly containing insecticides and fungicides) suitable for the actual situation of medicinal Crataegi Fructus based on the survey of the pesticides of the Crataegi Fructus base,combined with the blind screening results of the LC-ESI-MS/MS pesticide screening platform established by the research team in the early stage. Then,the pesticide residues in medicinal Crataegi Fructus from Shandong,Hebei,Henan,Shanxi,and Liaoning( main cultivation areas) were analyzed. The samples were pretreated by the modified Qu ECh ERS method,i.e.,extracted with acetonitrile-water( 9 ∶1),purified by PSA,C_(18),GCB,silica gel. The detection of pesticides was performed by LC-MS/MS. The ion source was ESI with positive scanning mode,and the linearity of 11 kinds of pesticides in the range of 5-300 μg·kg~(-1) was acceptable( R~2>0. 996 9). All the recoveries of pesticides were within 70. 02%~(-1)12. 0% in the low,medium and high levels,with RSD≤17%. The results showed that the detection rate of carbendazim,chlorpyrifos and difenoconazole is 79%,82%,56%,respectively. Besides,the prohibition pesticide carbofuran were detected in some of the batches,indicating the security risk. This study provides methodological references and basic data for risk assessment of Crataegi Fructus and government regulation.

Content Determination of 7 Components of Water Decoction of Compatibility of Crataegi Fructus and Rhei Radix et Rhizoma Before and After Stir-frying with Wine / 中国中医药信息杂志

Objective To establish a content determination method for 7 components of water decoction of compatibility of Crataegi Fructus and Rhei Radix et Rhizoma before and after stir-frying with wine; To provide a reference for the clinical compatibility of Rhei Radix et Rhizoma and Crataegi Fructus. Methods HPLC was performed on the column of Thermo-C18 endcapped column (250 mm×4.6 mm, 5 μm). The mobile phase was 0.1% phosphoric acid water-acetonitrile system and a linear gradient elution was used. The column temperature was 25 ℃, and the injection volume was 10 μL. The detection wavelength was set at 254 nm. Results The seven markers including gallic acid, chlorogenic acid, aloe-emodin, rheinic acid, rheum emodin, chrysophanic acid and emodin monomethyl ether, reached baseline separation. The linearity was good, and the average recovery was 104.2%, 100.3%, 99.8%, 102.8%, 103.5%, 98.5%, and 99.5%. RSDs of 7 components were all less than 2.7%. The method showed high precision, stability and reproducibility. After prepared with wine, gallic acid content increased in Rhei Radix et Rhizoma, while aloe emodin, rhein, emodin, chrysophanol and physcion decreased to varying degrees. The content of chlorogenic acid increased in water decoction of compatibility of Crataegi Fructus Rhei Radix et Rhizoma. Conclusion The method can be used for the content determination of 7 components of water decoction of the compatibility of Rhei Radix et Rhizoma and Crataegi Fructus before and after stir-frying with wine, providing references for clinical application.

Lipid regulating effect of mixture fermentation products of Crataegi Fructus, Alismatis Rhizoma, and Cassiae Semen with Monascus purpureus on hyperlipidemic rats / 中草药

Objective: To explore the lipid regulating effect of fermentation product of Crataegi Fructus, Alismatis Rhizoma, and Cassiae Semen on hyperlipidemic rats. Methods: SD rats were fed with high fat diet and established as hyperlipidemia animal model, the subjects were divided into six groups: control group, model group, positive control group, fermentation product prevention group, red yeast rice group, and fermentation product treatment group. After four weeks continuous oral administration, the effect of different medicine samples on serum triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) of hyperlipidemic rats were observed to assess the regulation effect. Results: Four weeks' continuous oral administration could regulate the TC, TG, LDL-C levels significantly lower (P < 0.001) and HDL-C level significantly increased (P < 0.05) in the prevention group compared with the model group. After modeling, continuous administration for four weeks, compared with the levels before giving medicines, TC, TG, LDL-C levels of positive control group and fermentation product treatment group were significantly regulated lower (P < 0.05, 0.01) and HDL-C level significantly increased (P < 0.05, 0.01). while the only TC and LDL-C levels in red yeast rice group showed significant effect. Conclusion: Product of lipid-lowering traditional Chinese medicines (TCMs) treated by solid-state fermentation with Monascus purpureus could effectively inhibit the formation of foodborne hyperlipidemia, as well as control and regulate hyperlipidemia, and synergistic effect also appeared between ingredients from TCMs and lovastatin.

Research on dissolution rule of main active ingredients in compatibility process of Compound Xuezhining / 中草药

Objective To study the content change of six main active ingredients (orange diclofenac, rubrofusarin-6-O-β-D- gentiobioside, emodin, stilbene glucoside, nuciferine, and quercetin) in the aqueous extract under different compatibility conditions of Compound Xuezhining (CX), using Kohonen maps model for data analysis of traditional Chinese medicine with obvious change in content, to further explore how the compatibility of TCM affect the dissolution characteristic of ingredient. Methods Through different combinations of different herbs in CX, the solubility of the six active ingredients was investigated by UPLC method before and after different compatibility modes; Analyzed the data and chosen Nelumbinis Folium which has significant quantity changes after compatibility as the object of research. The impacts of different compatibility modes on seven chemical compositions of Nelumbinis Folium were analyzed by supervised Kohonen maps model. Results The content change of six active ingredients after compatibility were different: the contents of nuciferine, orange diclofenac and emodin were mainly decreased, the content of stilbene glucoside was decreased, the content of quercetin was increased, and the content of rubrofusarin-6-O-β-D-gentiobioside showed no obvious regularity; When combined with Fructus Crataegi Pinnatifidae, seven chemical constituents in Nelumbinis Folium changed greatly and most of which were positively affected. Conclusion The effects of different compatibility modes on the solubility of active components are different in Chinese herbal formulae CX, and the content of chemical constituents in Nelumbinis Folium are more influential when combined with Fructus Crataegi Pinnatifidae.

Extraction Technology Optimization for Organic Acids in Crataegi Fructus by Central Composite De-sign-response Surface Method / 中国药房

OBJECTIVE:To optimize the extraction technology for organic acids in Crataegi fructus. METHODS:Based on sin-gle factor test,using liquid-solid ratio,extraction time and ethanol volume fraction as independent variables,extraction yield of or-ganic acids as dependent variable,central composite design-response surface method was used to optimize the extraction technology of organic acids in Crataegi fructus. RESULTS:The optimal extraction technology was as follow as liquid-solid ratio of 18.5:1, adding 75% ethanol,reflux extraction twice,2.0 h each time. Average extraction yield of organic acids in verification test was 5.22%(RSD=2.70%,n=3),with 1.75% relative error of the predicted value(5.13%). CONCLUSIONS:Optimized extraction technology for organic acids in Crataegi fructus is simple,with good reproducibility and predictability.

Preparation process of lipid-lowering medicines by solid state fermentation of Crataegi Fructus, Alismatis Rhizoma, and Cassiae Semen mixed with monascus / 中草药

Objective: To obtain a new lipid-lowering product that combines the advantages of lipid-lowering Chinese materia medica (CMM) and statin drugs. Solid-state fermentation was studied using medicinal and edible CMM as media components, Lovastatin yielding monascus screened in our laboratory was used as strain and Lovastatin yielding was used as index. Methods: Solid-state fermentation was optimized by single factor and orthogonal test, qualitative and quantitative analysis was conducted of the product via TLC and HPLC. Results: The optimal fermentation conditions contained 100 g/solid medium in 500 mL flask bottling capacity, material thickness of 2.5 cm, 10 mL liquid seed of 48 h, 10% inoculation volume, incubating at 30℃, breaking up the medium on day 2, adding 24% sterile water on day 3, cooling to 25℃ and culturing for 18 d in total, and Lovastatin yielding was up to 5.127 mg/g. Fermentation product of CMM contained more components compared with that without medicines, Lovastatin yielding increased by 42.27%, but γ-aminobutyric acid yielding decreased by 17.89%. Contents of main active ingredients ursolic acid, 2,3-acetyl alisol-B, chrysophanol, and physcion were increased by 232.7%, 173.7%, 767.6%, and 888.4%. Conclusion: Active ingredients of hawthorn, alisma, and cassia are released into the products after fermentation and contents of lipid-lowering active ingredients were improved significantly, new active ingredients are also noticed. Therefore, the fermentation process of lipid-lowering medicines obtained in this experimental study has some practical values.

Prescription rules of preparations containing Crataegi Fructus in Chinese patent drug / 中国中药杂志

To analyze the prescription rules of preparations containing Crataegi Fructus in the drug standards of the People's Republic of China Ministry of Public Health-Chinese Patent Drug(hereinafter referred to as Chinese patent drug), and provide some references for clinical application and the research and development of new medicines. Based on TCMISS(V2.5), the prescriptions containing Crataegi Fructus in Chinese patent drug were collected to build the database; association rules, frequency statistics and other data mining methods were used to analyze the disease syndrome, common drug compatibility and prescription rules. There were a total of 308 prescriptions containing Crataegi Fructus, involving 499 kinds of Chinese medicines, 34 commonly used drug combinations, and mainly for 18 kinds of diseases. Drug combination analysis was done with "Crataegi Fructus-Citri Reticulatae Pericarpium" and "Crataegi Fructus-Poria" as the high-frequency herb pairs and with "stagnation" and "diarrhea" as the high-frequency diseases. The results indicated that the Crataegi Fructus in different herb pairs had a roughly same function, and its therapy effect was different in different diseases. The prescriptions containing Crataegi Fructus in Chinese patent drug had the effect of digestion, and they were widely used in clinical application, often used together with spleen-strengthening medicines to achieve different treatment effects; the prescription rules reflected the prescription characteristics of Crataegi Fructus for different diseases, providing a basis for its clinically scientific application and the research and development of new medicines.

Variation analysis on triterpene compounds in hawthorn from different regions / 中草药

Objective: To establish an HPLC method for the content determination of ursolic acid and oleanolic acid in hawthorn (Crataegi Fructus) from different regions, whose differences were compared. Methods: Waters Symmetry C18 column (4.6 mm× 250 mm, 5 μm) was used with the mobile phase of acetonitrile-methanol-0.5% ammonium acetate (69:12:19) at a flow rate of 1 mL/min. The detection wavelength was set at 210 nm and the column temperature was 30°C. To determine the content of hawthorn from 10 different regions, variance analysis and cluster analysis were carried out for the results. Results: The linear ranges of ursolic acid and oleanolic acid were 0.078-0.780 mg/mL(r = 0.9998, n = 6) and 0.0169-0.169 mg/mL (r = 0.9996, n = 6), respectively. The average recoveries of the two components were 102.6% and 100.2%, in accordance with the determination requirement in Chinese Pharmacopoeia 2010. Conclusion: This method is simple, quick, accurate, and has better reproducibility for the determination of triterpene compounds in hawthorn. And hawthorn from Chengde and Jiangsu provinces is better. It could be used as the origin of hawthorn in the prescription of Xin mai Capsule.

Influence of different heating degrees on color of Chinese materia medica pieces / 中草药

Objective: To study the relativity between the heating degree and color value of Chinese materia medica pieces. Methods: The effects of heating temperature and time on the color value of Gardeniae Fructus and Crataegi Fructus were investigated by spectrophotometric technique. Results: The color value curve of Gardeniae Fructus after processing under the mid-and high-heating showed the better rules that the stationary phase of L* and δE* appeared in 7 min and stationary phase of a* and b* appeared in 4 min. However, the color value curve of Crataegi Fructus showed the stationary phase of L*, a*, b*, and δE* appeared in 5 min after low-heating process. After the mid-heating process, the stationary phase of L* and δE* appeared in 6 min, stationary phase of a* and b* values appeared in 5 and 8 min, respectively. After the high-heating process, the stationary phase of L* and δE* appeared in 8 min and the stationary phase of a* and b* value appeared in 5 min. Conclusion: There is the correlation of processing degree between the measurement results and the perusal observation, and the result of color measurement could provide the reference for confirming optimum parameters of processing technique.