Study on the mechanism of Fructus Forsythiae and Folium Nelumbinis on acute pharyngitis based on network pharmacology

Objective:To explore the mechanism of Lianqiao-Heye(Fructus Forsythiae and Folium Nelumbinis,FF-FN)drug pair in treating acute pharyngitis(AP)by network pharmacology.Methods:The chemical components and potential therapeutic targets of FF-FN drug pair were obtained from TCMSP firstly,and databases GeneCards,DrugBank,OMIM and PharmGKB were utilized to get the genes related to AP and the intersection of the results was obtained.Then Cytoscape software was used to construct drug-component-target network diagram to screen out the key compounds.Protein interaction network(PPI)was established using String database,and the core targets were screened by CytoNCA topology analysis.R language software was used for GO biological function analysis and KEGG pathway analysis.Finally,the docking verification of key compounds and core target molecules was carried out by AutoDock software.Results:A total of 38 active compounds and 917 potential therapeutic targets were obtained from FF-FN drug pair,while a total of 1534 targets were screened out for AP,among which,the number of intersection targets was 117.7 core targets were screened out from PPI core network,with JUN,TP53,CXCL8 and RELA included.A total of 2487 biological processes were involved in GO enrichment analysis,and 157 related pathways were screened out by KEGG.Based on results of molecular docking verification,the key compounds such as quercetin,luteolin and wogonin in FF-FN drug pair were proved capable of binding to the core targets and a good affinity was shown.Conclusion:FF-FN pair can intervene AP through multiple targets and multiple pathways,including PI3KAkts signaling pathway,AGE-RAGE signaling pathway and IL-17 signaling pathway.The combination of quercetin,luteolin,wogonin,kaempferol andβ-sitosterol with JUN,RELA,MAPK1,TNF and MYC can possibly be one of the mechanisms regarding to the therapeutic effect.

Preparation of Rosmarinic Acid Injection from Folium perillae

[Objective] To extract the effective component rosmarinic acid from Folium perillae,study preparation technology and quality control of rosmarinic acid injection and provide test basis for hemorheological drug development and clinic application.[Method]The coarse powder of Folium perillae(40 μm)was lixiviated with hot water and acidized to make aqueous extracts.Rosmarinic acid from the aqueous mixture was extracted with Ethyl Acetate.And Et Ac was evaporated to obtain primary product.The injection was purified by pH adjustment.Concentration of rosmarinic acid in the injection was detected by high performance liquid chromatography(HPLC).Contents of tannin,resin and oxalate were eliminated according to the Chinese Pharmacopeia(2005 edition).Stability,irritation,haemolyticus,LD50 and thermogenic substance of this injection were also detected.[Result]The content of rosmarinic acid in Folium perillae injection is 2.81 mg/ml,and the content of LD50 was 406.82 mg/kg.This injection was consistent with the stipulation of Chinese Pharmacopeia(2005 edition).[Conclusion]The preparation technology of this experiment was reasonable.The stable qualities of the prepared injection meet the injection requirement.

Hypoglycemic Effect of Forsythia suspensa Leaves on Diabetic Mice

[Objective] This study aimed to investigate the hypoglycemic effect of Forsythia suspensa leaves in diabetic mice. [Method] The model of hyperglycemia mice were obtained by injecting streptozotocin intraperitoneally to study the effect of F. suspensa leaves on lowering blood sugar of hyperglycemia mice and normal ani- mals. [Result] F. suspensa leaves could antagonize the streptozotocin-reducing mice hyperglycemia and significantly reduce the fasting blood glucose level of diabetic mice. [Coaclusion] F. suspensa leaves exhibits a good therapeutic effect on diabetic mice and has a good development prospect.

A Semi-Quantitative Analysis of Essential Micronutrient in Folium Lycii Using Laser-Induced Breakdown Spectroscopy Technique

Abstract In this paper, the capabilities of laser-induced breakdown spectroscopy （LIBS） for rapid analysis to multi-component plant are illustrated using a 1064 nm laser focused onto the surface of folium lycii. Based on homogeneous plasma assumption, nine of essential micronutrients in folium lycii are identified. Using Saha equation and Boltzmann plot method electron density and plasma temperature are obtained, and their relative concentration （Ca, Mg, A1, Si, Ti, Na, K, Li, and Sr） are obtained employing a semi-quantitative method.

Antiviral Activity of the Effective Monomers from Folium Isatidis Against Influenza Virus in Vivo

In order to evaluate the anti-influenza virus activity of the effective monomer from Folium Isatidis (FI) in vivo,we established mice model with viral pneumonia and divided them into 3 different dose groups,then observed their lung indexes,pulmonary pathological changes,pulmonary virus hemagglitination titers,living time and death rates.The results showed that the monomer could reduce the pulmonary index from 2.64 to 1.93,1.63 and 1.40 (P<0.01) and decrease the hemagglitination titer from 1.15 to 0.84,0.70 and 0.59 (P<0.01).In addition,different groups of FI could significantly lessen the mortality rate from 100% to 30%,25% and 15%,and prolong the living time from 5.1d to 6.5d,8.4d and 8.9d respectively(P<0.01).The high dose (75 mg/kg/d) has the similar effect with 100 mg/kg/d dose of virazole(P>0.05),and more effective than 200 mg/kg/d dose of antiviral liquor (P<0.05).

Potential therapeutic effects and applications of Eucommiae Folium in secondary hypertension

Eucommiae Folium(EF),a traditional Chinese medicine,has been used to treat secondary hypertension,including renal hypertension and salt-sensitive hypertension,as well as hypertension caused by thoracic aortic endothelial dysfunction,a high-fat diet,and oxidized low-density lipoprotein.The antihypertensive components of EF are divided into four categories:flavonoids,iridoids,lignans,and phenylpropanoids,such as chlorogenic acid,geniposide acid and pinoresinol diglucoside.EF regulates the occurrence and development of hypertension by regulating biological processes,such as inhibiting inflammation,regulating the nitric oxide synthase pathway,reducing oxidative stress levels,regulating endothelial vasoactive factors,and lowering blood pressure.However,its molecular antihypertensive mechanisms are still unclear and require further investigation.In this review,by consulting the relevant literature on the antihypertensive effects of EF and using network pharmacology,we summarized the active ingredients and pharmacological mechanisms of EF in the treatment of hypertension to clarify how EF is associated with secondary hypertension,the related components,and underlying mechanisms.The results of the network pharmacology analysis indicated that EF treats hypertension through a multicomponent,multi-target and multi-pathway mechanism.In particular,we discussed the role of EF targets in the treatment of hypertension,including epithelial sodium channel,heat shock protein70,rhoassociated protein kinase 1,catalase,and superoxide dismutase.The relevant signal transduction pathways,the ras homolog family member A(RhoA)/Rho-associated protein kinase(ROCK)and nicotinamide adenine dinucleotide phosphate(NADPH)oxidase/eNOS/NO/Ca^(2+)pathways,are also discussed.

Multidisciplinary strategies to enhance therapeutic effects of flavonoids from Epimedii Folium:Integration of herbal medicine,enzyme engineering,and nanotechnology

Flavonoids such as baohuoside I and icaritin are the major active compounds in Epimedii Folium(EF)and possess excellent therapeutic effects on various diseases.Encouragingly,in 2022,icaritin soft capsules were approved to reach the market for the treatment of hepatocellular carcinoma(HCC)by National Medical Products Administration(NMPA)of China.Moreover,recent studies demonstrate that icaritin can serve as immune-modulating agent to exert anti-tumor effects.Nonetheless,both production efficiency and clinical applications of epimedium flavonoids have been restrained because of their low content,poor bioavailability,and unfavorable in vivo delivery efficiency.Recently,various strategies,including enzyme engineering and nanotechnology,have been developed to increase productivity and activity,improve delivery efficiency,and enhance therapeutic effects of epimedium flavonoids.In this review,the structure-activity relationship of epimedium flavonoids is described.Then,enzymatic engineering strategies for increasing the productivity of highly active baohuoside I and icaritin are discussed.The nanomedicines for overcoming in vivo delivery barriers and improving therapeutic effects of various diseases are summarized.Finally,the challenges and an outlook on clinical translation of epimedium flavonoids are proposed.

ClCRY2 facilitates floral transition in Chrysanthemum lavandulifolium by affecting the transcription of circadian clock-related genes under short-day photoperiods

Plants sense photoperiod signals to confirm the optimal flowering time.Previous studies have shown that Cryptochrome2(CRY2)functions to promote floral transition in the long-day plant(LDP)Arabidopsis;however,the function and molecular mechanism by which CRY2 regulates floral transition in short-day plants(SDPs)is still unclear.In this study,we identified a CRY2 homologous gene,ClCRY2,from Chrysanthemum lavandulifolium,a typical SDP.The morphological changes in the C.lavandulifolium shoot apex and ClFTs expression analysis under SD conditions showed that adult C.lavandulifolium completed the developmental transition from vegetative growth to reproductive growth after eight SDs.Meanwhile,ClCRY2 mRNA exhibited an increasing trend from 0 to 8 d of SD treatment.ClCRY2 overexpression in wild-type(WT)Arabidopsis and C.lavandulifolium resulted in early flowering.The transcript levels of the CONSTANS-like(COL)genes ClCOL1,ClCOL4,and ClCOL5,and FLOWERING LOCUS T(FT)homologous gene ClFT1 were upregulated in ClCRY2 overexpression(ClCRY2-OE)C.lavandulifolium under SD conditions.The transcript levels of some circadian clock-related genes,including PSEUDO-REPONSE REGULATOR 5(PRR5),ZEITLUPE(ZTL),FLAVIN-BINDING KELCH REPEAT F-BOX 1(FKF1),and GIGANTEA(GI-1 and GI-2),were upregulated in ClCRY2-OE C.lavandulifolium,while the expression levels of other circadian clock-related genes,such as EARLY FLOWERING 3(ELF3),ELF4,LATE ELONGATED HYPOCOTYL(LHY),PRR73,and REVEILLE8(RVE8),were downregulated in ClCRY2-OE C.lavandulifolium under SD conditions.Taken together,the results suggest that ClCRY2 promotes floral transition by fine-tuning the expression of circadian clock-related gene,ClCOLs and ClFT1 in C.lavandulifolium under SD conditions.

Optimization of Ultrasound-assisted Enzymatic Polysaccharide Extraction from Turpiniae Folium Based on Response Surface Methodology

Objective To optimize the extraction procedure and obtain the maximum total polysaccharide yield from Turpiniae Folium.Methods Response surface methodology(RSM),combining Plackett–Burman design(PBD),steepest ascent method,and Box–Behnken design(BBD),was employed.The significant factors contributing to polysaccharide production were determined by PBD.The path of steepest ascent method was performed to rapidly reach the neighborhood of the optimum region.BBD and RSM were applied to further investigate the mutual interaction between the variables and to define the optimal values.Results The significant factors contributing to polysaccharide production were enzyme concentration,extraction time,and liquid-to-solid ratio.The optimal conditions for maximal TFP were:enzyme concentration of2.8%,extraction time of41min,and liquid-to-solid ratio of27mL/g.The experimental yield of3.08%was in good agreement with the model-predicted yield of3.1%under the optimized conditions.Conclusion This study was successfully applied to optimize the extraction conditions of Turpiniae Folium polysaccharide,which can contribute to its further production and application.

Microwave assisted-semi bionic extraction of lignan compounds from Fructus Forsythiae by orthogonal design

Microwave assisted-semi bionic extraction (MASBE) process for lignans from Fructus Forsythiae was studied. The influences of solvent pH value, microwave power, dosage of solvent and irradiation time were investigated. Optimum extracting parameters were determined by orthogonal experiments as follows: pH value of solvent at first extraction is 5.5-6.0, that at the second extraction is 7-8; microwave power is 700 W; mass ratio of Fructus Forsythiae to water is 1-12; irradiation time is 10 min, and extracting times is two. Under these optimal conditions, the yield of lignans reaches 0.364%. Compared with the conventional extraction methods, the MASBE process has the advantages of high extraction rate, high extraction selectivity.

Artemisiae Argyi Folium and its geo-authentic crude drug qi ai

Objective:To comprehensively review the literature relating to the use of Artemisiae Argyi Folium (AAF;the dried leaf of Artemisia argyi Lévl.et Vant.from the Asteraceae family),and qi ai (AAF grown in Qichun County and the adjacent areas in Hubei Province,China) in the field of traditional Chinese medicine.Furthermore,this study aimed to give results of the contents determination of three bioactive components in qi ai and AAF.Methods:The literature was reviewed to acquire relevant information regarding the quality and application of AAF and qi ai.In addition,the contents of the bioactive components (essential oil,total flavonoids and tannins) were determined in 29 AAF samples collected from China and Korea using the methods in the Chinese pharmacopoeia (2015) or determined by our experiments.Results:The dried leaf of A.argyi Lévl.et Vant.from the Asteraceae family is a mainstream commodity of AAF.Qi ai is a commercial type of AAF well-known for its good quality,which has been recorded in Chinese ancient literature.Modern research has showed that more intense heat with stronger penetration was generated in moxibustion therapy when qi ai moxa stick was burnt compared with general moxa stick,which led to a better curative effect.Our research also showed that the contents of essential oil,total flavonoids and tannin in qi ai were significantly higher than those of general AAF samples.Conclusion:The superior quality of qi ai compared with general AAF may be due to relatively higher contents of bioactive components.This study provides basic information for further study and utilization of AAF and qi ai.

Hepatoprotective Effects of Folium syringae Extracts Against Ethanolinduced Acute Liver Injury

The aim of the study was to investigate the hepatoprotective effects of Folium syringae(FS) extracts against ethanolinduced acute liver injury. Mice and primary hepatocytes were pretreated with FS extracts at different dosages before ethanol administration. Transaminases, glutathione S-transferase A1 level and hepatic biochemical indices(malondialdehyde, superoxide dismutase, glutathione and glutathione peroxidase) were determined. Pretreatment with FS extracts significantly inhibited the damage caused by ethanol and the hepatoprotective effects of FS were almost similar to Silymarin that was used to treat alcoholic liver injury. GSTA1 contents in all the FS extract-treated groups were significantly different from those in the ethanol-induced acute liver injury model group(p<0.01), and similar trends were observed in transaminases and hepatic indices level both in vitro and in vivo. The results showed that FS extracts had hepatoprotective effects against ethanol-induced injury. Those effects might be related to the enhancement of antioxidant capacity of liver cells, and FS extracts could reduce the release of liver GSTA1, which contributed to improve liver detoxification.

Associated Bacteriostasis Research of Coptidis Rhizoma-Folium Isatidis and Coptidis Rhizoma-Flos Poprli against Escherichia coli O2

[Objective] The paper was to explore the combined inhibitory effect of rhizoma coptidis-folium isatidis and rhizoma coptidis-flos poprli against Escherichia coli O2.[Method] Contrast test of single and associated bacteriostasis against known serotype E. coli O2 was conducted using microcheckerboard method.[Result] The MIC of rhizoma coptidis, folium isatidis and flos poprli were 1/8 extracting liquid, 1/8 extracting liquid and 1/2 extracting liquid, respectively. When combined with folium isatidis or flos poprli, the MIC of rhizoma coptidis was 1/8 extracting liq-uid or 1/16 extracting liquid compared with single use. When combined with rhizoma coptidis, the MIC of folium isatidis and flos poprli were 1/8 extracting liquid and 1/16 extracting liquid.[Conclusion] When rhizoma coptidis was combined with folium isatidis or flos poprli, the FIC values were 2 and 0.625, performing independent action and additive effect, respectively.

MBD protein recognizes flower control genes regulated by DNA methylation in Chrysanthemum lavandulifolium

Dynamic changes in DNA methylation regulate the expression of genes and play important roles especially in the flowering processes of higher plants.Methyl-CpG-binding domain protein could specifically recognize hypermethylated regions in the genome,thus MBD sequencing technology and CpG islands analysis of the sequences were used to identify candidate genes that were regulated by DNA methylation,in particular the flowering induction stage of Chrysanthemum lavandulifolium.MBD-seq identified 89 candidate genes which included 49 genes exhibiting changes in DNA methylation status during floral induction.Based on CpG islands analysis of the sequences,27 candidate genes were selected that may be regulated by DNA methylation.The expression levels of 30 candidate genes and nine key genes were determined by RT-PCR and qRT-PCR during floral induction(7D),four genes(ClFT,ClMET,DFL and ClWRKY21)were similarly up-regulated.Methylation-specific PCR analysis also indicated that there were changes in the DNA methylation status in the DFL and ClWRKY21.The changes in the DNA methylation status during the induction phase of flowering may lead to changes in gene expression.In this study,a set of genes were identified that are proposed to be involved in floral induction and two key genes were identified(DFL,ClWRKY21)that were regulated by DNA methylation during the flowering process of C.lavandulifolium.

Network Pharmacological Mechanism Research of Herba Drynariae Rhizoma-Epimedii Folium in Treating Osteoarthritis

Objective: To investigate the potential mechanism of Drynariae Rhizoma-Epimedii Folium in the treatment of osteoarthritis (OA) based on network pharmacology. Methods: The potential active constituents and targets of Drynariae Rhizoma-Epimedii Folium were screened through the traditional Chinese medicine (TCM) systems pharmacology database and analysis platform (TCMSP). Genecards database is used to find relevant targets of OA. The targets of “Drynariae Rhizoma-Epimedii Folium” were mapped to the targets of OA, and used Cytoscape software to build a “drug-ingredient-target-di- sease” regulatory network and protein protein interaction (PPI) network. R software was used to analyze the Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment of traditional Chinese medicine-disease targets. Results: Thirty-four effective ingredients and 130 traditional Chinese medicine-disease targets were screened out for the treatment of OA. The GO functions of traditional Chinese medicine-disease targets mainly included cytokine activity, cytokine receptor binding, nuclear receptor activity, transcription factor activity, proximal promoter DNA-binding transcription activator activity, DNA-binding transcription activator activity, phosphatase binding and so on. KEGG pathways involved in traditional Chinese medicine-disease targets mainly included TLR4 signaling pathway, TNF signaling pathway, IL-17 signaling pathway, MAPK signaling pathway, PI3K/AKT signaling pathway, apoptotic signaling pathway and so on. Conclusion: Network pharmacology may predict the multiple targets and multiple signaling pathways in Drynariae Rhizoma-Epimedii Folium treatment for OA, providing new ideas for future research.

A Novel Lignan Glucoside from Forsythia suspensa Vahl

从连翘果实中分离得到了五个单体化合物，根据理化性质和光谱分析鉴定为：对－羟基苯乙酸［ｐ－ｈｙｄｒｏｘｙｐｈｅｎｙｌａｃｅｔｉｃａｃｉｄ］（１），（＋）松脂素－β－Ｄ－葡萄糖甙［（＋）ｐｉｎｏｒｅｓｉｎｏｌ－β－Ｄ－ｇｌｕｃｏｓｉｄｅ］（２），（＋）表松脂素－４－β－Ｄ－葡萄糖甙［（＋）ｅｐｉｐｉｎｏｒｅｓｉｎｏｌ－４－β－Ｄ－ｇｌｕｃｏｓｉｄｅ］（３），（＋）松脂素单甲基醚－β－Ｄ－葡萄糖甙［（＋）ｐｉｎｏｒｅｓｉｎｏｌｍｏｎｏｍｅｔｈｙｌｅｔｈｅｒ－β－Ｄ－ｇｌｕｃｏｓｉｄｅ］（４），连翘甙［ｐｈｉｌｙｒｉｎ］（５）。其中３未见文献报道。

Analysis on Therapeutic Potential and Action Mechanism of Folium Pyrrosiae Based on Biolabel Pattern

[Objectives]The therapeutic potential and action mechanism of Folium Pyrrosiae were analyzed based on the biolabel pattern.[Methods]The chemical components of Folium Pyrrosiae were analyzed by liquid chromatography-mass spectrometry(LC-MS).Ten databases,including Pubchem,CTD,BindingDB,HERB,TCMIP,ETCM,SwissTargetPrediction,SuperPred webserver,TargetNet and SEA,were used in turn to retrieve the targets of related components,and key components were obtained according to the enrichment degree of targets.The obtained targets were imported into the STRING database to obtain PPI information and screen out core targets.The DAVID database was employed to analyze KEGG pathways of core targets and obtain key pathways.A key component-core target-key pathway network of Folium Pyrrosiae was constructed by Cytoscape3.10.1 software.The obtained KEGG pathways were input into the CTD database to predict corresponding diseases,and discussion and analysis were carried out.[Results]Ten key components,30 potential targets and 10 key pathways were screened out,and they participated in many diseases,of which five diseases were mainly analyzed.[Conclusions]Folium Pyrrosiae had the characteristic of multi-component,multi-target and multi-pathway synergistic effect in the treatment of lung cancer,type 2 diabetes,atherosclerosis,liver cancer,prostate cancer and other diseases,and the therapeutic potential and action mechanism of Folium Pyrrosiae were analyzed through the biolabel pattern.This study provides a research basis for further developing new functions of Folium Pyrrosiae.

Quality Standard for Euonymi Grandiflorus Caulis and Folium

[Objectives]To establish the quality standard for Euonymi Grandiflorus Caulis and Folium.[Methods]Qualitative identification was performed by methods of origin identification,microscopic identification and thin layer chromatography(TLC);the moisture,total ash,acid-insoluble ash,and extract were detected using the method in the Chinese Pharmacopoeia(2020 Edition),and the contents of quercetin and kaempferol were determined by High Performance Liquid Chromatography(HPLC).[Results]The leaves of Euonymi Grandiflorus were subleathery,narrow and long elliptic or narrow obovate with cyme,and capsules were often with narrow wing ridge.Characteristics of microscopic identification were significant.Calcium oxalate cluster crystals,cubic crystals,inlaid parenchyma cells,starch granules,stomata,and fiber bundles could be observed.A TLC method for identification of Euonymi Grandiflorus Caulis and Folium was established.The contents of water,total ash,acid-insoluble ash were not higher than 13%,9%,12%,respectively;the extract were no less were not higher than 13%,9%,12%and 17%in ten batches of samples,respectively;the sum of quercetin and kaempferol were no less than 0.20%.[Conclusions]The quality standard of Euonymi Grandiflorus Caulis and Folium were established through the research.This method is accurate,specific,and reproducible and can be used for the quality control of Euonymi Grandiflorus Caulis and Folium.

Prediction of the mechanisms of liver injury of Epimedii Folium by network pharmacology and validation in HepaRG Cells

Objective:EpimediiFolium(EF),a traditional Chinese medicinal material,has the effect of tonifying kidney Yang,strengthening bones and treating rheumatism.However,its clinical applications are limited by its drug-induced liver injury(DILI)effects and the underlying mechanisms have not been elucidated.Methods:Active EF compounds were obtained from the TCMSP database and their targets predicted in Targetnet.Next,DILI-targets were obtained from CTD,Genecards and Digsee databases.Protein-protein interactions of EF DILI-targets were determined using STRING and hub targets identified via topological analyses.Then,hub targets were subjected to GO and KEGG pathway enrichment analyses.Finally,HepaRG cells were used for further validation of molecular mechanisms.Results:Fifty seven active compounds and 164 targets that interacted with these active compounds were identified with Sagittatoside A,icariside I,and Icariin being the best active compounds.Enrichment analysis revealed the PI3K/Akt and NF-kB signaling pathways to be markedly enriched.Molecular docking revealed that Sagittatoside A,icariside I and Icariin had good binding activities to RAC1,PTGS2,and NOS3.Validation analysis in HepaRG cells revealed that Epimedium flavonoids upregulated RAC1,PTGS2 and NOS3 levels.Conclusion:Our findings show that EF induces oxidative stress,inflammation,and apoptosis via PI3K/Akt and NF-kB signaling pathways,and provides a basis for more in-depth studies on EF-induced DILI.

A new forsythenside from Forsythia suspensa

A new forsythenside, 4-hydroxy-4-[8-[[1-[（4-hydroxyphenyl）acetyl]-β-D-glucopyranosyl-6-]oxy]ethy1]-2,5-cyclohexadien-1- one, named forsythenside F, was isolasted from the plant of Forsythia suspensa （Thunb.） Vahl. The structure of the new compound was established on the basis of various spectroscopic analysis, including ^1H NMR, ^13C NMR, 2D NMR techniques （HMBC and HSQC）, and HR-ESI-MS.