Study on the Antipyretic and Anti-inflammatory Mechanism of Shuanghuanglian Oral Liquid Based on Gut Microbiota-Host Metabolism.

Nowadays, there has been increased awareness that the therapeutic effects of natural medicines on inflammatory diseases may be achieved by regulating the gut microbiota. Shuanghuanglian oral liquid (SHL), the traditional Chinese medicine preparation, has been shown to be effective in clearing heat-toxin, which is widely used in the clinical treatment of respiratory tract infection, mild pneumonia, and common cold with the wind-heat syndrome. Yet the role of gut microbiota in the antipyretic and anti-inflammatory effects is unclear. In this study, a new strategy of the 16S rRNA gene sequencing and serum metabolomics that aims to explore the role of SHL in a rat model of the systemic inflammatory response induced by lipopolysaccharide would be a major advancement. Our results showed that the gut microbiota structure was restored in rats with inflammation after oral administration of SHL, thereby reducing inflammation. Specifically, SHL increased the relative abundance of Bacteroides and Faecalibacterium and decreased the abundance of Bifidobacterium, Olsenella, Aerococcus, Enterococcus, and Clostridium in the rat model of inflammatory disease. Serum metabolomic profile obtained by the orbitrap-based high-resolution mass spectrometry revealed significant differences in the levels of 39 endogenous metabolites in the inflammatory model groups, eight metabolites of which almost returned to normal levels after SHL treatment. Correlation analysis between metabolite, gut microbiota, and inflammatory factors showed that the antipyretic and anti-inflammatory effects of SHL were related to the recovery of the abnormal levels of the endogenous metabolites (N-acetylserotonin and 1-methylxanthine) in the tryptophan metabolism and caffeine metabolism pathway. Taken together, these findings suggest that the structural changes in the gut microbiota are closely related to host metabolism. The regulation of gut microbiota structure and function is of great significance for exploring the potential mechanism in the treatment of lipopolysaccharide-induced inflammatory diseases with SHL.

A feasibility study on improving the non-invasive detection accuracy of bottled Shuanghuanglian oral liquid using near infrared spectroscopy.

The ultimate goal of the study is to present a strategy to improve the accuracy of near-infrared spectroscopy detection of Shuanghuanglian oral liquid in glass bottles without damaging the primary packaging. we adopted the multi-position spectral modeling (MPSM) method to correct the spectral variation caused by the difference of bottle and measuring position, so as to improve the measurement accuracy and find the best site combination for measuring Shuanghuanglian oral liquid. Baicalin, total flavonoids and soluble solid contents were considered as the quality indicators of the oral liquid, and partial least squares (PLS) models were employed for the single-position and multi-position spectra, respectively. The root mean square error of the validation set (RMSEP) of the optimum multi-position models are 0.7412 mg/mL for baicalin, 1.1259 mg/mL for total flavonoids and 0.9491% for soluble solids contents. Compared with the traditional single-position spectral modeling method (SPSM method), MPSM method improved the prediction accuracy of baicalin, total flavonoids and soluble solid contents by 26.84%, 31.97% and 58.14% respectively. The results showed that the MPSM method can improve the measurement accuracy of bottled oral liquid and is an effective method to eliminate the uncertainty of measurement conditions.

Effects of Shuanghuanglian oral liquids on patients with COVID-19: a randomized, open-label, parallel-controlled, multicenter clinical trial.

We conducted a randomized, open-label, parallel-controlled, multicenter trial on the use of Shuanghuanglian (SHL), a traditional Chinese patent medicine, in treating cases of COVID-19. A total of 176 patients received SHL by three doses (56 in low dose, 61 in middle dose, and 59 in high dose) in addition to standard care. The control group was composed of 59 patients who received standard therapy alone. Treatment with SHL was not associated with a difference from standard care in the time to disease recovery. Patients with 14-day SHL treatment had significantly higher rate in negative conversion of SARS-CoV-2 in nucleic acid swab tests than the patients from the control group (93.4% vs. 73.9%, P = 0.006). Analysis of chest computed tomography images showed that treatment with high-dose SHL significantly promoted absorption of inflammatory focus of pneumonia, which was evaluated by density reduction of inflammatory focus from baseline, at day 7 (mean difference (95% CI), -46.39 (-86.83 to -5.94) HU; P = 0.025) and day 14 (mean difference (95% CI), -74.21 (-133.35 to -15.08) HU; P = 0.014). No serious adverse events occurred in the SHL groups. This study illustrated that SHL in combination with standard care was safe and partially effective for the treatment of COVID-19.

Effects of Shuanghuanglian oral liquids on patients with COVID-19: a randomized, open-label, parallel-controlled, multicenter clinical trial / 医学前沿

We conducted a randomized, open-label, parallel-controlled, multicenter trial on the use of Shuanghuanglian (SHL), a traditional Chinese patent medicine, in treating cases of COVID-19. A total of 176 patients received SHL by three doses (56 in low dose, 61 in middle dose, and 59 in high dose) in addition to standard care. The control group was composed of 59 patients who received standard therapy alone. Treatment with SHL was not associated with a difference from standard care in the time to disease recovery. Patients with 14-day SHL treatment had significantly higher rate in negative conversion of SARS-CoV-2 in nucleic acid swab tests than the patients from the control group (93.4% vs. 73.9%, P = 0.006). Analysis of chest computed tomography images showed that treatment with high-dose SHL significantly promoted absorption of inflammatory focus of pneumonia, which was evaluated by density reduction of inflammatory focus from baseline, at day 7 (mean difference (95% CI), -46.39 (-86.83 to -5.94) HU; P = 0.025) and day 14 (mean difference (95% CI), -74.21 (-133.35 to -15.08) HU; P = 0.014). No serious adverse events occurred in the SHL groups. This study illustrated that SHL in combination with standard care was safe and partially effective for the treatment of COVID-19.

Anti-SARS-CoV-2 activities in vitro of Shuanghuanglian preparations and bioactive ingredients.

Human infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19) and there is no cure currently. The 3CL protease (3CLpro) is a highly conserved protease which is indispensable for CoVs replication, and is a promising target for development of broad-spectrum antiviral drugs. In this study we investigated the anti-SARS-CoV-2 potential of Shuanghuanglian preparation, a Chinese traditional patent medicine with a long history for treating respiratory tract infection in China. We showed that either the oral liquid of Shuanghuanglian, the lyophilized powder of Shuanghuanglian for injection or their bioactive components dose-dependently inhibited SARS-CoV-2 3CLpro as well as the replication of SARS-CoV-2 in Vero E6 cells. Baicalin and baicalein, two ingredients of Shuanghuanglian, were characterized as the first noncovalent, nonpeptidomimetic inhibitors of SARS-CoV-2 3CLpro and exhibited potent antiviral activities in a cell-based system. Remarkably, the binding mode of baicalein with SARS-CoV-2 3CLpro determined by X-ray protein crystallography was distinctly different from those of known 3CLpro inhibitors. Baicalein was productively ensconced in the core of the substrate-binding pocket by interacting with two catalytic residues, the crucial S1/S2 subsites and the oxyanion loop, acting as a "shield" in front of the catalytic dyad to effectively prevent substrate access to the catalytic dyad within the active site. Overall, this study provides an example for exploring the in vitro potency of Chinese traditional patent medicines and effectively identifying bioactive ingredients toward a specific target, and gains evidence supporting the in vivo studies of Shuanghuanglian oral liquid as well as two natural products for COVID-19 treatment.

Can network pharmacology identify the anti-virus and anti- inflammatory activities of Shuanghuanglian oral liquid used in Chinese medicine for respiratory tract infection?

INTRODUCTION: Shuanghuanglian (SHL) oral liquid is a well-known traditional Chinese medicine preparation administered for respiratory tract infections in China. However, the underlying pharmacological mechanisms remain unclear. The present study aims to determine the potential pharmacological mechanisms of SHL oral liquid based on network pharmacology. METHODS: Network pharmacology-based strategy including collection and analysis of putative compounds and target genes, network construction, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and Gene Ontology (GO) enrichment, identification of key compounds and target genes, and molecule docking was performed in this study. RESULTS: A total of 82 bioactive compounds and 226 putative target genes of SHL oral liquid were collected. Of note, 28 hub target genes including 4 major hub target genes: estrogen receptor 1 (ESR1), nuclear receptor coactivator 2 (NCOA2), nuclear receptor coactivator 1 (NCOA1), androgen receptor (AR) and 5 key compounds (quercetin, luteolin, baicalein, kaempferol and wogonin) were identified based on network analysis. The hub target genes mainly enriched in pathways including PI3K-Akt signaling pathway, human cytomegalovirus infection, and human papillomavirus infection, which could be the underlying pharmacological mechanisms of SHL oral liquid for treating diseases. Moreover, the key compounds had great molecule docking binding affinity with the major hub target genes. CONCLUSION: Using network pharmacology analysis, SHL oral liquid was found to contain anti-virus, anti-inflammatory, and "multi-compounds and multi-targets" with therapeutic actions. These findings may provide a valuable direction for further clinical application and research.

Combination of western medicine and Chinese traditional patent medicine in treating a family case of COVID-19.

In December 2019, an outbreak of novel coronavirus (2019-nCoV) occurred in Wuhan, Hubei Province, China. By February 14, 2020, it has led to 66 492 confirmed patients in China and high mortality up to ~2.96% (1123/37 914) in Wuhan. Here we report the first family case of coronavirus disease 2019 (COVID-19) confirmed in Wuhan and treated using the combination of western medicine and Chinese traditional patent medicine Shuanghuanglian oral liquid (SHL). This report describes the identification, diagnosis, clinical course, and management of three cases from a family, suggests the expected therapeutic effects of SHL on COVID-19, and warrants further clinical trials.

Clinical Observation of Shuanghuanglian Oral Liquid Combined with Cefuroxime Axetil in the Treatment of Bacterial Respiratory Tract Infection / 中国药房

OBJECTIVE:To observe the clinical efficacy and safety of Shuanghuanglian oral liquid combined with cefuroxime axetil in the treatment of bacterial respiratory tract infection. METHODS:184 patients with bacterial respiratory tract infection were randomly divided into test group and control group. Test group was orally given 250 mg Cefuroxime axetil tablet,twice a day+20 ml Shuanghuanglian oral liquid,3 times a day. Control group was only given Cefuroxime axetil tablet (the same dosage as test group). The treatment course for 2 groups was 2 weeks. The cough duration,body temperature recovery time and runny nose disap-pearing time before and after treatment and bacterial clearance rate in 2 groups were observed,clinical efficacy and incidence of ad-verse reactions were recorded. RESULTS:After treatment,the total effective rate in test group was significantly higher than control group,cough duration,body temperature recovery time and runny nose disappearing time were significantly shorter than control group,the differences were statistically significant(P0.05). CONCLUSIONS:The efficacy of Shuanghuanglian oral liquid combined with ce-furoxime axetil is superior to cefuroxime axetil alone in the treatment of bacterial respiratory infections,with similar safety.

Selective flocculation processing technology of chitosan complex flocculant on Shuanghuanglian aqueous extract / 中草药

Objective: To optimize the selective flocculation processing technology of chitosan complex flocculant on Shuanghuanglian aqueous extract. Methods: The impacts of preparation technology, the dosage of complex flocculant, the time of flocculation, and the performance index of chitosan on the flocculating effect of Shuanghuanglian aqueous extract were examined by using the retention rates of chlorogenetic acid and baicalin, as well as the elimination rate of tannin as observation indicators. Results: The selective flocculation processing technology on Shuanghuanglian aqueous extract was optimized as follows: the bentonite was loaded by chitosan after it was burned for 4 h at 450°C, the dosage of complex flocculant was 60 g/L, the flocculating time was 24 h, and the best performance index of chitosan is 95% in deacetylation, and 60 cps in viscosity. Conclusion: The complex flocculant made of high temperature modified bentonite loaded by chitosan shows the good selectivity in flocculation capacity. The flocculating effect of this method on Shuanghuanglian aqueous extract is remarkably better than that in the one documented in Chinese Pharmacopoeia.

In vivo and in vitro correlation of quality control components in Shuanghuanglian Oral Liquid / 中草药

Objective: To establish the in vitro and in vivo HPLC fingerprint of Shuanghuanglian Oral Liquid (SOL). Methods: The chromatographic fingerprint was obtained with Welchrom C18 column (150 mm × 4.6 mm, 5 μm); The mobile phase was methanol-0.2% phosphoric acid aqueous solution, with gradient elution and segmentation variable wavelength determination; The column temperature was maintained at 30°C; The injection volume was 10 μL. Results: Among 10 batches of samples prepared by single and different preparation methods, the overall similarity evaluation was taken using the Chromatographic Fingerprint Similarity Software for Chinese Materia Medica (Version 2004A), and the average similarities were all above 0.93. The better in vivo and in vitro correlation of the index constituents in SOL prepared by the method of Pharmacopoeia of People's Republic of China was obtained. Conclusion: The method had good precision, stability, and repeatability. The standard chromatographic fingerprint established has a better representativity, and the method provides the new ideas for establishing the new evaluation system of Chinese materia medica oral preparation.

Quick Identification of Shuanghuanglian Oral Liquid and Yinhuang Oral Liquid by AOTF-near Infrared Spectroscopy Principal Component Analysis / 中国药房

OBJECTIVE:To apply AOTF-near infrared spectroscopy for quick identification of Shuanghuanglian oral liquid and Yinhuang oral liquid.METHODS:Yinhuang oral liquid and Shuanghuanglian oral liquid samples of different batches from different manufacturers were collected for collection of their near infrared spectra.The qualitative analysis models of the two kinds of oral liquid were established respectively using principal component analysis.Meanwhile,the established models were used to analyze and validate each other.RESULTS:The established qualitative analysis models of Shuanghuanglian oral liquid and Yinhuang oral liquid could be applied to validate and identify each other correctly.CONCLUSION:The method was proved to be a simple,rapid new analytical technique with low cost,and it can be used for a quick identification of Shuangh-uanglian oral liquid and Yinhuang oral liquid.

Validation of Microbial Limit Tests of Shuanghuanglian Oral Liquid / 中国药房

OBJECTIVE:To validate the microbial limit tests of Shuanghuanglian oral liquid.METHODS:The recovery rates of 4 kinds of test organisms including E.coli treated by Shuanghuanglian oral liquid were detected by routine method,culture medium dilution method,and membrane-filter procedure respectively.RESULTS:Shuanghuanglian oral liquid was found to be of bacteriostatic action.The contamination of solution couldn't be detected by routine method.The recovery rate of the test organisms detected by membrane-filter procedure was higher than by culture medium dilution method,but the recovery rates in both methods were all up to the requirement.CONCLUSION:Membrane-filter procedure should be considered preferentially in performing microbial limit test for Shuanghuanglian oral liquid.

Determination of Chlorogenic Acid, Baicaliin, Forsythin and Wogonin in Shuanghuanglian Oral Liquid by HPLC / 中国中医药信息杂志

Objective To establish a HPLC method for the determination of chlorogenic acid, baicaliin, forsythin and wogonin in Shuanghuanglian oral liquid. Methods The column was VP-ODS C18 (250 mm?4.6 mm, 5 ?m). The mobile phase consisted of acetonitrile-0.2% phosphoric acid with gradient elution. The flow rate was 1.0 mL/min and the detection wavelength was at 278 nm. Results The calibration curves were linear within the range of 0.44～6.60 ?g (r=0.999 1) for chlorogenic acid, 0.52～6.18 ?g (r=0.999 1) for baicaliin, 0.20～2.04 ?g (r =0.999 3) for forsythin (r =0.999 3) and 0.13～1.76 ?g (r =0.999 1) for wogonin, respectively. The average recovery of them were 97.0% (RSD=1.1%), 98.98% (RSD=1.1%), 103.55% (RSD = 1.1%) and 96.49% (RSD = 1.1%), respectively. Conclusion The method is simple, practicable, accurate and rapid. It can be applied to content determination of chlorogenic acid, baicaliin, forsythin and wogonin in Shuanghuanglian oral liquid.

Quick identification of Shuanghuanglian Oral Liquid / 中成药

AIM: To establish a new method for quick identifying Shuanghuanglian Oral Liquid(Flos Lonicerae japonicae,Radix Scutella riae,Fructus Forsythiae) by AOTF-Near infrared spectroscopy. METHODS: Near infrared spectroscopy combined with principal component analysis. RESULTS: The model SHL-C of Shuanghuanglian Oral Liquid could distinguish Shuanghuanglian Oral Liquid from Yinhuang Oral Liquid and other 9 kinds of these(oral) liquids. CONCLUSION: The method is quick,simple and low cost assay technique and can be used to identification of Shnanghuanglian Oral Liquid.

Determination of forsythin in Shuanghuanglian Oral Liquid by RP-HPLC / 中成药

Objective: To establish a method for determination of forsythin in Shuanghuanglian Oral (Flos Lonicerae, Radix Scutellariae, Fructus Forsythiae, etc.) Methods: After processing by RP SPE (reverse phase solid phase extraction), the sample solution was measured by RP HPLC. The chromatographic conditions were: Prodigy ODS (3) (150?4.6mm,5?m) column as analytic column; MeOH H 2O HAc(40∶60∶1,V/V) as mobile phase; detection wavelength at 227nm; column temperature at 35?C ; flow rate at 1.0mL?min -1 . Results: The linear range of forsythin was 0.1～2.0?g, r =0.9997. The average recovery was 102.2%, RSD =0.61%( n =6). Conclusion: The method is simple, accurate, reproducible and can be used to enhance the quality control of this drug.