Clinical Effect of Modified Gegen Qinlian Decoction Combined with Western Medicine on Children with Rotavirus Enteritis with Damp-Heat Syndrome

[Objectives] To explore the clinical effect of Modified Gegen Qinlian Decoction combined with western medicine on children with rotavirus enteritis with damp-heat syndrome and its influence on myocardial enzymes.

Screening of Anti-inflammatory Active Ingredients from Gancao Qinlian Extract and Study of Its Efficacy

[Objectives]To establish the chromatographic fingerprint of Gancao Qinlian Extracts(GQE)and reveal the possible material basis of the anti-inflammatory effect of GQE by the correlation analysis between the fingerprint chromatographic peaks of different components of GQE and its anti-inflammatory activity.[Methods]Ultra-performance liquid chromatography(UPLC)was used to detect the different ingredients of GQE to establish its chromatographic fingerprint and analyze the differences among the three medicine components;LPS stimulated RAW264.7 cells to construct an inflammatory cell model.The NO secretion of cells was detected by the Griess method.ELISA was used to detect the changes in TNF-αand IL-10 contents.RT-qPCR tested the mRNA expression levels of TNF-αand IL-10.Grey relational analysis was carried out by combining fingerprint chromatographic peak data and anti-inflammatory activity data.[Results]The GQE fingerprint was established,34 fingerprint characteristic peaks were calibrated,and 33 related chromatographic peaks were screened out.The corresponding chromatographic peaks in the three components were obtained,and the content of the components was calculated;the anti-inflammatory results showed that the content of NO,TNF-α,and the expression of TNF-αmRNA in the high and medium-dose groups of GQE were significantly lower than those in the blank group(P<0.01).The NO content and TNF-αmRNA expression in the high-dose group of GQE I was considerably lower than those in the blank group(P<0.01).The secretion of NO,TNF-α,and the expression of TNF-αmRNA in the high,medium,and low dose groups of GQE II were significantly lower than those in the blank group(P<0.01);the results of grey relational analysis showed that the correlation degree of the three components was GQE II>GQE>GQE I,and the characteristic fingerprint peaks 12,15,22,23,28,31,33 may be closely related to the anti-inflammatory effect.[Conclusions]The best component of the anti-inflammatory effect in GQE is water-soluble component,and its main components are flavonoids and alkaloids.These components can alleviate cellular inflammatory damage by inhibiting the excessive secretion of NO and reducing the expression of TNF-αmRNA.

The potential role of Gegen Qinlian decoction in the treatment of coronavirus disease 2019 based on network pharmacology and validation of lipopolysaccharide-induced macrophages

Background:Coronavirus disease 2019(COVID-19)is caused by severe acute respiratory syndrome coronavirus 2,which has led to deaths and currently lacks an efficient treatment.Despite studies suggesting the potential of the Gegen Qinlian decoction(GQD)in preventing COVID-19,comprehensive analyses of its anti-COVID-19 potential are still lacking.Methods:GQD treatment was evaluated for its efficacy in ameliorating the early stage(24 hours)of lipopolysaccharide(LPS)-induced cytokine storm in vivo.Additionally,target genes of GQD were co-analyzed with COVID-19 signature genes to identify key ingredients and their pathways.Validation was also conducted using an LPS-induced macrophage model.Results:GQD treatment effectively ameliorated the early stage of LPS-induced cytokine storm in vivo.Key ingredients such as quercetin were found to be involved in multiple pathways,including inflammation,immunity,oxidative stress,cell proliferation,and apoptosis,through the AGE-RAGE signaling pathway and IL-17 signaling pathway.In the LPS-induced macrophage model,quercetin inhibited macrophage polarization(M1)and the secretion of inflammatory factors(IL-6,TNF-α,IL-17A).Conclusions:Our results indicate that GQD can be utilized in the treatment of cytokine storm induced by COVID-19 and has the potential to treat COVID-19 by suppressing the COVID-19 signature genes and macrophage polarization.

Mechanism of Gegen Qinlian Decoction in Treating Type 2 Diabetes Mellitus Complicated with NAFLD Based on Network Pharmacology

[Objectives]To explore the mechanism of Gegen Qinlian Decoction in treating type 2 diabetes mellitus(T2DM)complicated with non-alcoholic fatty liver disease(NAFLD)by analyzing the effective components of Gegen Qinlian Decoction.[Methods]TCMSP database was used to analyze the active components of Gegen Qinlian Decoction,and pubchem and Swiss ADME databases were also used to predict drug targets,extract T2DM complicated with NAFLD targets from OMIM and Genecards databases.Venny plot was drawn to obtain intersection targets,and finally Cytoscape was used to make core target maps and drug-target-disease network maps.Using DAVID and Metascape database to analyze the intersection targets,the gene ontology information of Go and KEGG was obtained.Microbial informatics technology was used to visualize GO,and Cytoscape was used to make drug-target-disease network map-enrichment pathway map.[Results]The network pharmacological analysis showed that Gegen Qinlian Decoction acted on the key targets of type 2 diabetes mellitus complicated with non-alcoholic fatty liver disease,such as ALB and ALT1,through many components,and achieved the purpose of treating this disease.The chemical constituents of the drug include formononetin,5-hydroxyisomucronulatol-2,5-2-O-glucoside,cholesteryl laurate,isoliquiritigenin,etc.[Conclusions]This study provides a new idea and theoretical support for future drug research and clinical practice.

Gegen Qinlian decoction enhances immunity and protects intestinal barrier function in colorectal cancer patients via gut microbiota

BACKGROUND We previously showed,using the Traditional Chinese Medicine System Pharmacology Database,that Gegen Qinlian decoction(GQD)had a direct antitumor effect,and was combined with programmed cell death protein(PD)-1 inhibitors to treat microsatellite stable(MSS)tumor-bearing mice.However,the effect of GQD on patients with colorectal cancer(CRC)is not clear.AIM To determine the therapeutic mechanism of GQD in improving immune function,reducing inflammation and protecting intestinal barrier function.METHODS Seventy patients with CRC were included in this study:37 in the control group and 33 in the treatment group.The proportions of CD4+T,CD8+T,natural killer(NK),NKT and T regulatory cells were measured by flow cytometry.Levels of the cytokines tumor necrosis factor(TNF)-α,interferon(IFN)-γ,interleukin(IL)-2,IL-6,IL-10 and serotonin(5-hydroxytryptamine;5-HT)in serum were assessed by enzyme-linked immunosorbent assay(ELISA).The expression of zonula occludens(ZO)-1,occludin,nuclear factor(NF)-κB and TNF-αin tumor and normal tissues was measured by immunohistochemistry.The composition of gut microbiota from patients in the treatment group was assessed using 16S rDNA analysis.RESULTS There were no adverse events in the treatment group.The proportion of CD4+T cells and NKT cells in the post-treatment group was significantly higher than that in the pre-treatment and control groups(P<0.05).The level of TNF-αin the posttreatment group was significantly lower than that in the pre-treatment and control groups(P<0.05).The concentration of 5-HT in the post-treatment group was significantly lower than that in the pre-treatment group(P<0.05).The expression of ZO-1 and occludin in tumor tissues in the treatment group was significantly higher than that in the control group(P<0.05).The expression of ZO-1 in normal tissues of the treatment group was significantly higher than that in the control group(P=0.010).Compared with the control group,expression of NF-κB and TNF-αin tumor tissues of the treatment group was significantly decreased(P<0.05).Compared with the pre-treatment group,GQD decreased the relative abundance of Megamonas and Veillonella.In addition,GQD increased the relative abundance of Bacteroides,Akkermansia and Prevotella.CONCLUSION GQD enhances immunity and protects intestinal barrier function in patients with CRC by regulating the composition of gut microbiota.

Determination of Adulteration Dye Auramine O in Gegen Qinlian Tablets by HPLC-MS/MS

[Objectives]This study aimed to establish a method for determination of adulteration dye auramine O in Gegen Qinlian tablets by HPLC-MS/MS.[Methods]The instrument conditions were as follows:column,Agilent Eclipse plus C18(2.1 mm×100 mm,1.8μm);mobile phase,acetonitrile-0.01 mol/L ammonium acetate;gradient elution;flow rate,0.3 mL/min;column temperature,35℃;injection volume,2μL;ion source,electrospray ionization source(ESI);and scanning mode,multiple reaction monitoring(MRM).[Results]Methodological validation shows that auramine O had a good linear relationship in the range of 0.0934-1.869 pg(r=0.9999),and the average recovery rate was 94.38%(RSD=3.60%,n=9).[Conclusions]The method established is accurate,convenient,precise,reproducible,and stable,with good linear relationship,and it can be used determine the content of adulteration dye auramine O in Gegen Qinlian tablets.

Administration of modified Gegen Qinlian decoction for hemorrhagic chronic radiation proctitis: A case report and review of literature

BACKGROUND Hemorrhagic chronic radiation proctitis(CRP) is a common late complication of irradiation of the pelvis and seriously impairs life quality. There is no standard treatment for hemorrhagic CRP. Medical treatment, interventional treatment, and surgery are available, but they are limited in their applications due to nondefinite efficacy or side effects. Chinese herbal medicine(CHM), as a complementary or alternative therapy, may provide another option for hemorrhagic CRP treatment.CASE SUMMARY A 51-year-old woman with cervical cancer received intensity-modulated radiation therapy and brachytherapy with a total dose of 93 Gy fifteen days after hysterectomy and bilateral adnexectomy. She received six additional cycles of chemotherapy with carboplatin and paclitaxel. Nine months after radiotherapy treatment, she mainly complained of 5-6 times diarrhea daily and bloody purulent stools for over 10 d. After colonoscopy examinations, she was diagnosed with hemorrhagic CRP with a giant ulcer. After assessment, she received CHM treatment. The specific regimen was 150 mL of modified Gegen Qinlian decoction(GQD) used as a retention enema for 1 mo, followed by replacement with oral administration of 150 mL of modified GQD three times per day for 5 mo. After the whole treatment, her diarrhea reduced to 1-2 times a day. Her rectal tenesmus and mild pain in lower abdomen disappeared. Both colonoscopy and magnetic resonance imaging confirmed its significant improvement. During treatment,there were no side effects, such as liver and renal function damage.CONCLUSION Modified GQD may be another effective and safe option for hemorrhagic CRP patients with giant ulcers.

Study on the molecular mechanism of Gegen Qinlian decoction in the treatment of influenza based on network pharmacology

Objective:The study shows that Gegen Qinlian decoction(GGQL)is feasible in the treatment of influenza.It is explained from traditional Chinese medicine theory and modern pharmacology,and the formula is tested in vitro.The results show that Gegen Qinlian decoction has a good inhibitory effect on influenza virus.Methods:Search TCMSP database to get the components and targets of GGQL,and construct the compound target network.Then search OMIM database and Disgenet database to obtain influenza genes,and the common genes were obtained by VennDiagram.STRING database was used to construct gene function association network.Finally,David database was used to analyze GO enrichment and pathway enrichment.Results:146 ingredients and 128 targets were obtained,588 genes were obtained.46 common genes were obtained by VennDiagram,and import the common genes into the STRING database and construct the gene function association network,then the network information data was imported into Cytoscape software for topology analysis.Finally,13 enriched GO clusters and 7 enriched pathway clusters were obtained by David database,and construct the targets-GO-pathway network.Conclusion:Network pharmacology of GGQL reveal that the target and the pathway of the drug was closely related to influenza,and the results of this study have been verified with published experimental studies,which shows that GGQL has not only theoretical basis for the treatment of influenza,but also in vitro experiments support this conclusion.

Simultaneous Determination of Effective Components of Gegen Qinlian Tablets by HPLC

[Objectives]To establish a HPLC method for the simultaneous determination of 14 characteristic components in Gegen Qinlian tablets and the content of Gegen Qinlian tablets in 11 manufacturers.[Methods]A Inertsil ODS-3(250 mm×4.6 mm,5μm)reversed-phase high performance liquid chromatography column was used with acetonitrile-0.02 mol/L ammonium acetate+0.03%triethylamine solution(adjusted with glacial acetic acid to pH of 4.3)as the mobile phase for gradient elution at a flow rate of 1.0 mL/min.The determination wavelength of 8 components(puerarin,daidzin,liquiritin,baicalin,wogonoside,daidzein,ammonium glycyrrhizinate,baicalein)was 250 nm;the determination wavelength of wogonoside was 280 nm;the determination wavelength of 5 components(epierberine,jatrorrhizine hydrochloride,coptisine,palmatine hydrochloride and berberine hydrochloride)was 346 nm.[Results]There was a good linear relationship between the injection volume of puerarin,daidzin,liquiritin,baicalin,wogonoside,epierberine,jatrorrhizine hydrochloride,coptisine,daidzein,palmatine hydrochloride,ammonium glycyrrhizinate,berberine hydrochloride,baicalein and wogonin and peak area(r>0.9990)in the range of 146.26-5850.24,24.13-965.04,18.45-738.00,79.18-3167.32,9.57-382.80,4.76-190.40,2.57-102.80,13.41-536.40,10.60-424.00,11.33-453.22,12.08-483.20,46.73-1869.25,20.28-811.20,12.11-484.50 ng,respectively;the average recovery rates(n=6)were 2.18%,1.79%,1.81%,1.68%,2.27%,2.13%,1.96%,1.07%,0.93%,0.61%,2.92%,0.77%,2.79%and 0.62%,respectively;the precision,repeatability and stability were good,and the RSD was less than 3%.This method was used to determine 16 batches of Gegen Qinlian tablets produced by 11 enterprises.Wogonoside was not detected in a lot of samples,but 14 components were detected for other enterprises,but the content was different.[Conclusions]The method was accurate and feasible and could be used for the overall quality control of this variety.

Explanation of Gegen Qinlian Decoction Syndrome and Its Clinical Application

Gegen Qinlian Decoction is one of the commonly used classical prescriptions,which consists of fourherbs:Gegen(Puerariae Lobatae Radix),Huangqin(Scutellariae Radix),Huanglian(Coptidis Rhizoma),and Zhigancao Glycyrrhizae Radix et Rhizoma Praeparata cum Melle.The clinical application experience of Gegen Qinlian Decoction is as follows:first,according to Zhongjing Zhang's original text,heat distressing the large intestine with unresolved exterior syndrome is theoriginal meaningof Gegen Qinlian Decoction syndrome.In clinical practice,bacillary dysentery,gastrointestinal cold,etc.,belong to simultaneous exterior and interior disease,which are completely consistent with the original meaning of this prescription;second,the clinical digestive system diseases with exuberant heat of the large intestine as the core manifestation cannot be treated merely based on the exterior syndrome;third,due to the interior-exterior relationship between the lung and the large intestine and between the meridians,the use of Gegen(Puerariae Lobatae Radix),Huangqin(Scutellariae Radix),and Huanglian(Coptidis Rhizoma)can make the source of body clear,and the waste qi can be released and thepore can be easily opened,so this prescription can also be used for exogenous fever;fourth,with multidimensional comprehensive understanding of the syndrome,pathogenesis,symptoms and pharmacology,Gegen Qinlian Decoction has also been further applied to the treatment of hypertension,diabetes,etc.

廉州话概说

廉州话是合浦县的主体方言,也是粤语钦廉片的两个代表之一。主要语音特点是:古全浊声母今读塞音、塞擦音的85%送气,15%不送气。心母94%读边擦音。古清音去声合并于古浊音平声,归阳平。浊入声调高化,54%合并于清入。同音字汇收字4千个,词语举例收常用词语155条,语法例句55条。

灵山话纪略

灵山话是灵山县的主体方言,讲灵山话的占全县人口的71.31%。灵山话属于粤语钦廉片,最突出的特点是,古全浊声母今读塞音和塞擦音的字基本送气,果合一有三分之一的字与遇合三、蟹合一合流读[u],假开三大半读[i],没有撮口呼,遇合一、遇合三唇音读[?u],止开三、止合三微韵非组主要读[oi],阴平读低降调。同音字汇收字3900个,词语举例收常用词语134个,语法例句收生活语句50句。

浦北福旺背山话概说

浦北县内使用人口最多的一种粤语次方言称为背山话。其突出特点是:古全浊声母今读塞音、塞擦音的绝大部分读送气清音,果摄不论开合主要读[u],假开三主要读[i],没有撮口呼,遇摄不论洪细都读[u],咸深摄少数字、曾开三及梗摄大部分字读前鼻音韵尾。词语举例收词155个,语法例句收57个,同音字汇收字4000个。

Double-blinded,randomized clinical trial of Gegen Qinlian decoction pinpoints Faecalibacterium as key gut bacteria in alleviating hyperglycemia

Background:Accumulating evidence suggests that metabolic disorders,including type 2 diabetes mellitus(T2DM),can be treated with traditional Chinese medicine formulas,such as the Gegen Qinlian decoction(GQD).This study elucidates the mechanisms by which gut microbes mediate the anti-diabetic effects of GQD.Methods:We conducted a double-blind randomized clinical trial involving 120 untreated participants with T2DM.During the 12-week intervention,anthropometric measurements and diabetic traits were recorded every 4 weeks.Fecal microbiota and serum metabolites were measured before and after the intervention using 16S rDNA sequencing,liquid chromatography-mass spectrometry,and Bio-Plex panels.Results:Anti-diabetic effects were observed in the GQD group in the human trial.Specifically,glycated hemoglobin,fasting plasma glucose,and two-hour postprandial blood glucose levels were significantly lower in the GQD group than in the placebo group.Additionally,Faecalibacterium was significantly enriched in the GQD group,and the short-chain fatty acid levels were higher and the serum inflammation-associated marker levels were lower in the GQD group compared to the placebo group.Moreover,Faecalibacterium abundance negatively correlated with the levels of serum hemoglobin,fasting plasma glucose,and pro-inflammatory cytokines.Finally,the diabetes-alleviating effect of Faecalibacterium was confirmed by oral administration of Faecalibacterium prausnitzi(DSMZ 17677)in T2DMmousemodel.Conclusions:GQD improved type 2 diabetes primarily by modulating the abundance of Faecalibacterium in the gut microbiota,alleviating metabolic disorders and the inflammatory state.

Simultaneous analysis of 12 bioactive constituents in Gegen Qinlian pill by HPLC-DAD

In this study, a novel and simple high performance liquid chromatography with diode array detection （HPLC-DAD） method for the simultaneous qualitative and quantitative determination of 12 bioactive components in Gegen Qinlian pill was developed. The separation was performed on a Kromasil C18 column （250 mm×4.6 ram, 5.0 μm） by gradient elution with acetonitrile and 0.02 mol/L ammonium acetate （containing 0.3% triethylamine, and adjusted pH to 4.3 using 1% glacial acetic acid） as the mobile phase at a flow rate of 0.7 mL/min. Three different detection wavelengths （250, 280, 346 nm） were set at the maximum UV absorption wavelengths of these components. Twelve components （puerarin, daidzin, baicalin, wogonoside, liquiritin, berberine, palmatine, jatrorrhizine, glycyrrhizin, baicalein, wogonin and daidzein） were identified and determined using the developed method. All calibration curves showed good linear regression （r〉0.9995） within tested ranges. The injection precision, intra-day precisions and analysis repeatability were evaluated with the RSD values, which were no more than 0.97%, 1.69% and 1.71%, respectively. The recoveries were ranged from 97.4% to 100.2% with RSD values less than 1.87%. This readily available, low-cost and reliable HPLC-DAD method would improve the quality control of Gegen Qinlian pill.

Antidiabetic Effects of Gegen Qinlian Decoction via the Gut Microbiota Are Attributable to Its Key Ingredient Berberine

Gegen Qinlian Decoction(GQD),a traditional Chinese medicine(TCM)formula,has long been used for the treatment of common metabolic diseases,including type 2 diabetes mellitus.However,the main limitation of its wider application is ingredient complexity of this formula.Thus,it is critically important to identify the major active ingredients of GQD and to illustrate mechanisms underlying its action.Here,we compared the effects of GQD and berberine,a hypothetical key active pharmaceutical ingredient of GQD,on a diabetic rat model by comprehensive analyses of gut microbiota,short-chain fatty acids,proinflammatory cytokines,and ileum transcriptomics.Our results show that berberine and GQD had similar effects on lowering blood glucose levels,modulating gut microbiota,inducing ileal gene expression,as well as relieving systemic and local inflammation.As expected,both berberine and GQD treatment significantly altered the overall gut microbiota structure and enriched many butyrate-producing bacteria,including Faecalibacterium and Roseburia,thereby attenuating intestinal inflammation and lowering glucose.Levels of short-chain fatty acids in rat feces were also significantly elevated after treatment with berberine or GQD.Moreover,concentration of serum proinflammatory cytokines and expression of immune-related genes,including Nfkb1,Stat1,and Ifnrg1,in pancreatic islets were significantly reduced after treatment.Our study demonstrates that the main effects of GQD can be attributed to berberine via modulating gut microbiota.The strategy employed would facilitate further standardization and widespread application of TCM in many diseases.

Effect of Puerarin on the Pharmacokinetics of Baicalin in Gegen Qinlian Decoction (葛根芩连汤) in Mice

Objective： To study the pharmacokinetics of puerarin（PUE） in Gegen Qinlian Decoction（葛根芩连汤, GQD）, and the effects of PUE dosage variations on the pharmacokinetics of baicalin（BAL） in mice. Methods： GQD is composed of the concentrated granules of four Chinese herbs. Three dosages with different levels of PUE, including GQD, GQD co-administered with PUE, and GQD co-administration with two times the amount of PUE, were used to research the pharmacokinetics of PUE and BAL in mice. The indirect competitive enzyme-linked immunosorbent assay（ic ELISA） methods based on an anti PUE-monoclonal antibody（MAb） and BAL-MAb were employed to determine the concentration of PUE and BAL in mice blood. Results： After the co-administration of GQD with PUE, the area under the curves（AUC0-14 h） of PUE increased 2.8 times compared with GQD. At the dose of GQD co-administration at two times that of PUE, the AUC0-14 h of PUE was almost equal to that of GQD co-administration of PUE, showing non-linear pharmacokinetics. The AUC0-48 h of BAL showed a good dose-related increase of PUE（r=0.993） in the range from 100 to 300 mg/kg, indicating that PUE dramatically affects the absorption of BAL in mice. There was no significant difference in the other pharmacokinetic parameters, such as the first time of maximum concentration（Tmax）, the second Tmax, or the mean residence time. Conclusions： The ic ELISA methods were successfully applied to pharmacokinetic studies of PUE and BAL in GQD in mice. The dosage variability of PUE of the main ingredient in GQD affects its own pharmacokinetic characteristics and the absorption characteristics of BAL.

Comprehensive Metabolic Profiling of Modified Gegen Qinlian Decoction by Ultra-high-Performance Liquid Chromatography-Diode Array Detection-Q-Exactive-Orbitrap-Electrospray Ionization-Mass Spectrometry/Mass Spectrometry and Application of High-Performance

Objective:Gegen Qinlian decoction(GQD)is a classical traditional Chinese medicine formulation which has been used for almost 2000 years.At Guang'anmen Hospital,Beijing,a modified GQD version(m GQD)with seven instead of four herbal ingredients has been applied to treat Type 2 diabetes.Quality control is a crucial prerequisite for the therapeutic application of herbal medicines.For the identification of products derived from classical GQD,the Chinese Pharmacopeia requires the analysis of only three marker compounds.Because m GQD is a more complex mixture containing seven herbs and hundreds of constituents,the pharmacopoeia method for GQD is inadequate.Materials and Methods:A more comprehensive characterization of the formula's constituents has been developed using ultra-high-performance liquid chromatography-diode array detection(UHPLC-DAD)-Q-Exactive-mass spectrometry(MS)in electrospray ionization positive and negative mode.Moreover,a new method for the fingerprint analysis of m GQD via high-performance thin-layer chromatography(HPTLC)has been established.Results:Altogether,91 compounds have been assigned to their originating plants and 84 substances were identified either by comparison with authentic references or with data from the literature.The HPTLC method is based on the application of two different mobile phases and is able to detect both lipophilic and hydrophilic constituents of m GQD.Conclusions:The modified GQD was extensively characterized by UHPLC combined with DAD and Q-Exactive Orbitrap high-resolution MS detection,leading to the assignment and identification of compounds present in the decoction.In addition,a new method for the fingerprint analysis of the m GQD using HPTLC was established,which allows fast and simple identification of the herbal ingredients in the mixture.