Clinical Efficacy and Application Progress of Reduning Injection in Children with Viral Encephalitis

Viral encephalitis in children caused by multiple viral infections,as a common central nervous system disease in pediatrics,has a high incidence in China.Its clinical symptoms usually include infection,poisoning,intracranial hypertension,and disturbance of consciousness.In severe cases,it can even endanger the lives of patient children.At present,the clinical treatment of the viral encephalitis mainly uses traditional Chinese medicine preparations such as Reduning Injection.The efficacy is significant.It can not only relieve the uncomfortable symptoms in a short time,but also has low incidence of adverse reactions.In this study,the factors that cause viral encephalitis in children were analyzed,and the efficacy,pharmacological action and clinical application of Reduning Injection were comprehensively elaborated,to provide a reference for clinical doctors to rationally treat viral encephalitis in children.

Network pharmacology studies on Reduning injection in treatment of COVID-19

Background:Network pharmacology was used to explore the mechanism of the Chinese patent medicine Reduning injection in the treatment of corona virus disease 2019.Methods:The chemical constituents and targets of Qinghao(Artemisiae annuae herba),Jinyinhua(Lonicerae japonicae flos),Zhizi(Gardeniae fructus)in the Chinese patent medicine Reduning injection were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,and the target related to corona virus disease 2019 was searched in GeneCards database,then perform Venn analysis on the targets of corona virus disease 2019 and the Chinese patent medicine Reduning injection,to screen the compounds and targets of the Chinese patent medicine Reduning injection in the treatment of corona virus disease 2019.The String platform was used to construct the protein-protein interaction network,and key targets were screened.Cytoscape 3.5.1 was used to construct the active traditional Chinese medicine-chemical composition-target-disease network to screen the key active components,and the gene ontology function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed on the common target through DAVID(6.8)online analysis data tool to predict the mechanism of action.Results:Fifty-two active ingredients and 232 targets were selected from the Chinese patent medicine Reduning injection,including 44 targets related to corona virus disease 2019.A total of 310 gene ontology biological processes and 94 Kyoto Encyclopedia of Genes and Genomes signaling pathways were obtained,which were mainly involved in inflammation,viral infection,bacterial infection,immune response and substance metabolism,et al.Conclusion:The mechanism of the Chinese patent medicine Reduning injection in the treatment of corona virus disease 2019 may be related to antivirus,bacteriostasis,anti-inflammatory and antifebrile,immune regulation and metabolism regulation,et al.

The Mechanism of Reduning Injection in the Treatment of COVID-19 associated Myocardial Injury Based on Network Pharmacology and Molecular Docking

Objective:To explore the potential active components and mechanism of Reduning injection in the treatment of coronavirus disease 2019(COVID-19)associated myocardial injury by using molecular docking and network pharmacology.Methods:The main chemical constituents and molecular target of Reduning injection were collected from TCMID.Genes related to 2019 ncov were screened by genecards.Cytoscape software was used to construct and analyze the network.The active components of Reduning injection were molecularly docked with a new coronavirus hydrolase and its binding proteins,angiotensin converting enzyme II(ACE2)and transmembrane serine proteases(TMPRSSs).Results:There were 25 active ingredients and 198 drug targets in Reduning injection,43 targets proteins of coronavirus were excavated.Its main components have good binding ability with viral hydrolase and related binding proteins.Conclusion:Reduning injection may intervene the inflammatory response by multi-target and multi-component,inhibit the activity of coronavirus and its binding proteins ACE2 and TMPRSSs,and play a role in the treatment of new coronavirus pneumonia and myocardial injury.

Meta-analysis of the comparison between reduning injection and ribavirin injection for acute upper respiratory tract infection in children

Objective:To systematically evaluate the effectiveness and safety of reduning injection in acute upper respiratory tract infection of children.Methods:Use computer to search PubMed,the Cochrane Library,EMBASE,Web of Science,China National Knowledge Infrastructure(CNKI),Wanfang Database,Weipu database(VIP)and China Biomedical studies Service System(CBM)for randomized controlled trials(RCT)of acute upper respiratory tract infection in children,and the retrieval time is from their establishment to May,2022.After two researchers independently screened the studies,extracted data and evaluated the risk bias of studies,RevMan5.3 software was used to perform meta-analysis.Results:10 studies in total were included,involving 1466 patients.Results of the meta-analysis:total effective rate[RR=1.12,95%CI(1.07,1.16),P<0.00001],cure time[MD=-1.51,95%CI(-1.71,-1.32),P<0.00001],antipyretic time[MD=-1.32,95%CI(-1.72,-0.92),P<0.00001],disappearing time of nasal obstruction and nose running[MD=-1.04,95%CI(-1.45,-0.62),P<0.00001],disappearing time of cough[MD=-1.35,95%CI(-1.58,-1.13),P<0.00001],disappearing time of throat congestion[MD=-1.36,95%CI(-1.68,-1.03),P<0.00001],disappearing time of sore throat[MD=-1.55,95%CI(-1.74,-1.35),P<0.00001],the incidence of adverse reactions[RR=0.34,95%CI(0.20,0.57),P<0.0001].The differences between two groups in total effective rate,cure time,antipyretic time,disappearing time of nasal obstruction and nose running,disappearing time of cough,disappearing time of throat congestion,disappearing time of sore throat and the incidence of adverse reactions are all statistically significant(P<0.05).The sensitivity analysis demonstrates that three outcomes(total effective rate,cure time and disappearing time of sore throat)have unstable results,while others having stable results.Conclusion:Reduning injection is more effective than ribavirin injection in clinical,and it is better than ribavirin injection at relieving fever,nasal obstruction and nose running,cough and throat congestion.In addition,it’s safer in clinical than ribavirin injection.

Active Ingredients of Reduning Injection Maintain High Potency against SARS-CoV-2 Variants

Objective:To investigate the anti-coronavirus potential and the corresponding mechanisms of the two ingredients of Reduning Injection:quercetin and luteolin.Methods:A pseudovirus system was designed to test the efficacy of quercetin and luteolin to inhibit SARS-CoV-2 infection and the corresponding cellular toxicity.Luteolin was tested for its activities against the pseudoviruses of SARS-CoV-2 and its variants.Virtual screening was performed to predict the binding sites by Autodock Vina 1.1.230 and PyMol.To validate docking results,surface plasmon resonance(SPR)was used to measure the binding affinity of the compounds with various proteins of the coronaviruses.Quercetin and luteolin were further tested for their inhibitory effects on other coronaviruses by indirect immunofluorescence assay on rhabdomyosarcoma cells infected with HCoV-OC43.Results:The inhibition of SARS-CoV-2 pseudovirus by luteolin and quercetin were strongly dose-dependent,with concentration for 50%of maximal effect(EC_(50))of 8.817 and 52.98μmol/L,respectively.Their cytotoxicity to BHK21-hACE2 were 177.6 and 405.1μmol/L,respectively.In addition,luetolin significantly blocked the entry of 4 pseudoviruses of SARS-CoV-2 variants,with EC50lower than 7μmol/L.Virtual screening and SPR confirmed that luteolin binds to the S-proteins and quercetin binds to the active center of the 3CLpro,PLpro,and helicase proteins.Quercetin and luteolin showed over 99%inhibition against HCoV-OC43.Conclusions:The mechanisms were revealed of quercetin and luteolin inhibiting the infection of SARS-CoV-2 and its variants.Reduning Injection is a promising drug for COVID-19.

Antiviral effects of Reduning injection against Enterovirus 71 and possible mechanisms of action

The present study was designed to evaluate the protective effects of Reduning injection against Enterovirus 71(EV71) in Vero cells and in mice. The Vero cells were infected with 100 and 50 TCID50(50% tissue culture infective dose) of EV71, respectively. The inhibition of Reduning injection on cytopathic effect(CPE) was detected. Meanwhile, a mouse model produced by intraperitoneal EV71-infection(106 TCID50), was used to investigate the protective effects of Reduning injection. The total survival rate, living time, daily survival rate, weight ratio, and score for symptoms were examined. The viral loads in Vero cells and muscle tissues were detected using real-time PCR. Finally, the content of cytokines was analyzed by ELISA. In the Vero cells, 2.5 mg crude drug·m L-1 of Reduning injection inhibited CPE induced by EV71 infection. In the mice, 1.3 g crude drug·kg-1 of Reduning injection rescued death triggered by infection, in comparison with model group. Moreover, the survival rate, weight ratio, and clinical scores were also improved. The viral RNA copies in the Vero cells and the mice muscle tissues were reduced. Besides, the steep EV71-induced accumulations of TNF-α and MCP-1 were decreased by Reduning injection. In conclusion, Reduning injection showed promising protective effects against EV71 in Vero cells and in mice.

Anti-Inflammatory Effects of Reduning Injection(热毒宁注射液) on Lipopolysaccharide-Induced Acute Lung Injury of Rats

Objective： To evaluate the protective effects of Reduning Injection （热毒宁注射液, RDN）, a patent Chinese medicine, on lipopolysaccharide （LPS）-induced acute lung injury （ALl） in rats and its underlying mechanisms of action. Methods： Sixty male Sprague-Dawley rats were randomly divided into 6 groups, including normal control, model, dexamethasone （DEX, 5 mg/kg）, RDN-H （720 mg/kg）, RDN-M （360 mg/kg） and RDN-L （180 mg/kg） groups, with 10 rats in each group. Rats were challenged with intravenous injection of LPS 1 h after intraperitoneal treatment with RDN or DEX. At 6 h after LPS challenge, lung tissues and bronchoalveolar lavage fluid （BALF） were collected, and the number of inflammatory cells was determined. The right lungs were collected for histopathologic examination, measurement of gene and protein expressions, superoxide dismutase （SOD） and myeloperoxidase （MPO） activities. Results： In vivo pretreatment of RDN （360, 720 mg/kg） significantly reduced the weight of wet to dry （W/D） ratio of lung, protein content in BALF, and led to remarkable attenuation of LPS-induced histopathological changes in the lungs. Meanwhile, RDN enormously decreased BALF total inflammatory cells, especially neutrophil and macrophage cell numbers. Moreover, RDN increased SOD activity, inhibited MPO activity, alleviated LPS-induced tumor neurosis factor-o~ （TNF-o~） and inducible nitric oxide synthase （iNOS） expression in lung tissues. Furthermore, RDN （720 mg/kg） efficiently weakened nuclear factor- kappa B （NF- K B） gene and protein expression. Conclusion： Anti-inflammatory effects of RDN was demonstrated to be preventing pulmonary neutrophil infiltration, lowering MPO activity, TNF-oL and iNOS gene expression by inhibiting NF- K B activity in LPS-induced ALl.

A randomized controlled clinical study on treatment of hand, foot and mouth disease in children with Reduning injection

Hand, foot and mouth disease （HFMD） has become a major public health problem in China since 2008,which has been widely prevalent in children under 5 years of age. According to the data of the Ministry of Health, the number of cases with HFMD exceeded 1 million during the recent 4 years. The major etiologic agent of HFMD is enterovirus 71 and coxsackievirus A 16. The main clinical manifestations of the disease include fever, maculopapular skin vesicles on the hand, foot and buttock, oral herpes, ulcer; ation severe cases there can be central nervous system infection and/or pulmonary hemorrhage, which are mostly caused by EV71 and may be life-threatening with a fatality rate of 0.3 to 0.5 per thousand cases. There are no specific antiviral drugs, nor vaccine against EV71.~ The difficult problem of the treatment is how to reduce the severe cases and mortality. Reduning injection is a preparation of traditional Chinese herbal medicine composed of Qinghao （Artemisiae annuae）, Jinyinhua （Flos lonicerae） and Zhizi （Gardenia jasminoides）, which was listed among the therapies for HFMD in the 2010 guidelines of the Ministry of Health for diagnosis and treatment of HFMD.2 We now reported the results of our study on the efficacy of Reduning injection in treatment of 81 patients with HFMD. We selected 81 cases who were inpatients treated in 2010 in the Department of Infectious Diseases of Quanzhou Chidren＇s Hospital. The inclusion criteria were as follows： 1. Patients who met either the clinical diagnostic criteria for HFMD ordinary clinical type or the clinical diagnostic criteria for confirmed cases of HFMD （etiologically confirmed cases must account for more than 1/3 of the cases）. 2. The axillary temperature is _〉37.5~C, within 48 hours after onset of the disease; 3. Skin changes such as the rashes and vesicles occurred in no more than 48 hours; 4. Range of age is between 1 and 13 years; 5. Patients or their parents or guardians are willing to participate in the study and sign the informed consent forms. The exclusion criteria were as follows： 1. Patients who are complicated with the following primary diseases： congenital heart disease, chronic hepatitis, acute or chronic nephritis, diseases of hematologic system; 2. Patients who have hypersensitivity with history of allergy to the study drugs or other drugs; 3. Patients who were treated with western or traditional Chinese medicines other than the study medicines for HFMD by the time of the visit to the hospital; 4. Patientswho have been participated in other clinical trial for HFMD. The patients were centrally randomized （the random numbers were obtained from the web version of the Randomization System of Clinical Studies, Clinical Evaluation Center of Chinese Academy of Traditional Chinese Medicine, No. 200907001-3） into the following 3 groups： Reduning group （group 1）, 28 cases with 21 men and 7 women; age 12-72 （mean 29.2＋18.8） months; 24 were positive for EV71, 2 were positive for CoxA16 and 2 were negative for both; treated with intravenous infusion of Reduning （Jiangsu Kangyuan Pharmaceutical Corporation, Ltd., 10 ml in each vial, Batch number 100122） at a dose of 0.5 ml.kg-＇.d＂ added into 10% glucose solution for injection, once a day. Symptomatic treatment group （group 2）, 27 cases with 19 male and 8 female; age 12-96 （mean 28.7＋20.4） months; 19 were positive for EV71, 3 were positive for CoxA16 and 5 were negative for both; these patients were treated with intravenous infusion of vitamin C （0.5-1.0 g/d） and B6 （50-100 mg/d） added into 10% glucose solution for injection and ibuprofen （5-10 mg/kg per day, 6-8 hours repeat p.r.n.） or Aspisol （15-25 mg＇kg＂.d1） was used for antipyretic purpose. Antibiotics were added for patients who had increased peripheral blood leukocyte count. And Reduning plus symptomatic treatment group （group 3）, had 26 cases with 17 male and 9 female; age 12-84 （mean 30.4＋16.6） months; 18 were positive for EV71, 4 were positive for CoxA16 and 5 were negative for both viruses; these cases were treated with both Reduning and symptomatic treatments. No significant difference was found among the 3 groups in age, sex and etiological characteristics compared using chi-square test （P 〉0.05）. The numbers of patients who were positive for EV7I or CoxA 16 accounted for 75.3% and 11.1% for each group.

Reduning plus ribavirin display synergistic activity against severe pneumonia induced by H1N1 influenza A virus in mice

OBJECTIVE:To investigate synergistic effect of Reduning(RDN)injection plus ribavirin against severe pneumonia induced by H1 N1 influenza A virus in mice.METHODS:We established a mouse model of severe pneumonia induced by influenza A virus by infecting Balb/c mice with CA07 virus.We randomly assigned the infected mice into four groups,and treated them with normal saline(NS group),RDN(injection,86.6 mg/kg),ribavirin(injection,66.6 mg/kg)or double Ribavirin plus RDN group,the same dosage as used in the single treatments)for 5 d.Lung index and lung pathology were recorded or calculated in terms of the curative effective.Cytokines,NOD-like receptor family pyrin domain containing 3(NLRP3)inflammasome related protein including caspase-associated recruitment domain(CARD)domain Apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),caspase-1 and NOD-like receptor family,pyrin domain containing 3(NLRP3),and reactive oxygen species were simultaneously investigated.RESULTS:RDN plus ribavirin treatment,not RDN or ribavirin alone,provided a significant survival benefit to the influenza A virus-infected mice.The combination treatment protected the mice against severe influenza infection by attenuating the severe lung injury.The combined treatment also reduced the viral titers in mouse lungs and lung index,downregulated their immunocytokine levels,including IL-1βand IL-18,and down regulated the NLRP3,especially the transcription and translation of caspase-1.Meanwhile NS group had significantly higher reactive oxygen species(ROS)expression which could was dramatically reduced by the treatment of RDN plus ribavirin.CONCLUSION:Our study showed that RDN combined with ribavirin could protect the mice,and reduce the lung immunopathologic damage caused by severe influenza pneumonia.The mechanism could be that it reduced ROS produce and inhibited NLRP3 inflammasome activation so that mainly lower the downstream inflammatory cytokines IL-1βand IL-18.

Reduning injection for fever, rash, and ulcers in children with mild hand, foot, and mouth disease: a randomized controlled clinical study

OBJECTIVE： To assess the efficacy and safety of Re- duning injection for fever, rash, and ulcers in chil- dren with mild hand, foot, and mouth disease （HFMD）. METHODS： A stratified-block randomized, dou- ble-blind, parallel-controlled, and multicenter clini- cal trial was conducted with 360 patients in five hospitals across China： Quanzhou Children＇s Hospi- tal, Shijiazhuang No. 5 Hospital, Shanghai Public Health Centre, Hunan Provincial Children＇s Hospi- tal, and Kaifeng Children＇s Hospital. Patients were randomized into three groups with 120 in each. Group A was treated with Western Medicine, group B with Reduning injection, a Chinese herbal medi- cine, and group C with both Reduning injection and Western Medicine. Results were compared for treatment efficacy and safety on HFMD. The clinical outcomes were observed as follows： fever and on- set time of antifebrile effect （time to bring the body temperature down 〉0.5~C after medication）; cumu- lative time for fever recovery （body temperature re- covering to normal and lasting more than 24 h without medication）; cumulative time for rash dis- appearance （without new rashes or ulcers appear- ing and the original ones fading away）; and cumula- tive time for mouth ulcer disappearance. RESULTS： For the onset time of the antifebrile ef- fect, there was no statistical difference between groups A and B （P〉O.05） and groups B and C （P〉 0.05）. However, there was a statistical difference be- tween groups A and C （P〈0.05）, and the effect ingroup C was the best. For the cumulative time for rash disappearance, there was no statistical differ- ence between groups A and B （P〉0.05）. There were statistical differences between groups A and C, and groups B and C （P〈0.05）, and the effect in group C was the best. For the cumulative time for mouth ul- cers disappearance, there were no statistical differ- ences among the three groups （P〉0.05）. CONCLUSION： Reduning injection with Western Medicine for symptomatic treatment is most effec- tive for mild HFMD. No adverse reactions were ob- served.

Reduning Injection versus Neuraminidase Inhibitors in Treatment of Influenza:A Systematic Review and Meta-Analysis

Objective:To perform a systematic review to assess the effectiveness and safety of Reduning Injection versus neuraminidase inhibitors in treatment of influenza.Methods:The MEDLINE,Embase,the Cochrane Central Register of Controlled Trials(CENTRAL),Chinese Bio-medical Literature and Retrieval System(Sinomed),China National Knowledge Infrastructure Database(CNKI),China Science and Technology Journal Database(VIP),Wanfang Data Knowledge Service Platform and ClinicalTrails.gov were systematically searched from inception dates to May 2021 for randomized controlled trials(RCTs)exploring Reduning Injection alone or in combination with neuraminidase inhibitors in patients with influenza.Statistical analysis was performed using RevMan 5.4 and Stata 15.1.The qualities of the involved studies were assessed by the risk of bias according to the Cochrane handbook.The evidence quality of each outcome was evaluated by GRADEpro GDT.Results:Twelve trials with 1,460 patients were included.The included studies had a certain unclear or high risk of bias.Reduning Injection appeared to be more effective in shortening the fever clearance time(MD:-16.20 h,95%CI:-19.40 to-12.99,7 trials,814 patients,I^(2)=94%,very low certainty),fever alleviation time(MD:-4.09 h,95%CI:-4.22 to-3.96,3 trials,366 patients,I^(2)=0%,low certainty),cough alleviation time(MD:-21.34 h,95%CI:-41.56 to-1.11,2 trials,228 patients,I^(2)=89%,very low certainty),fatigue alleviation time(MD:-31.83 h,95%CI:-36.88 to-26.77,2 trials,270 patients,I^(2)=0%,low certainty),sore throat alleviation time(MD:-28.66 h,95%CI:-32.23 to-25.10,1 trial,150 patients,low certainty),and improving the total effective rate(RR:1.15,95%CI:1.06 to 1.25,10 trials,1,074 patients,I^(2)=76%,very low certainty).Besides,Reduning Injection seemed generally safe.Conclusions:This study provided low or very low evidence indicating Reduning Injection may be effective in the treatment of influenza and might be safe.Further rigorously designed studies are needed to confirm the effectiveness and safety of Reduning Injection and support it as a recommendation for influenza.

Two new terpenoids from Reduning Injection

Objective:To study the antipyretic and anti-inflammatory constituents from the active fraction of Reduning(RDN)Injection.Methods:In this study,the active fraction of RDN Injection was screened by the LPS-induced mouse endotoxin shock model.The chemical constituents were isolated by chromatography on HP-20 macroporous adsorptive resins,silica gel,ODS columns and reverse phase MPLC and HPLC repeatedly,and their structures were elucidated based on spectroscopic analysis(HR-ESI-MS,NMR,ECD)and chemical methods.Meanwhile,we evaluated the anti-inflammatory activities of the isolates by measuring their inhibitory effects on TNF-αproduction in LPS stimulated RAW 264.7 macrophages.Results:The 95%ethanol eluate of RDN Injection by the macroporous adsorption resin column was proved to be the antipyretic and anti-inflammatory active fraction of this injection.A novel iridoid,named jasminoide A(1),and a new guaiane sesquiterpenoid,named(1 R,7 R,8 S,10 R)-7,8,11-trihydroxy-4-guaien-3-one(2),were isolated from Reduning injection,and compound 2 can inhibit TNF-αproduction with IC50 values of 72.24μmol/L.Conclusion:In this study,two new terpenoids were isolated from Reduning Injection,and compound 2 showed inhibitory activity against LPS-activated TNF-αproduction in RAW 264.7 cells in vitro.

Reduning Injection prevents carrageenan-induced inflammation in rats by serum and urine metabolomics analysis

Objective:To elucidate the anti-inflammatory mechanism of Reduning Injection(RDN)by analyzing the potential biomarkers and metabolic pathways of the carrageenan-induced inflammatory model from the overall metabolic level.Methods:Rat inflammatory model was established by carrageenan.UPLC-Q-TOF/MS was used to detect and analyze changes of endogenous metabolites in the serum and urine of carrageenan-induced inflammatory rats.Combined with multivariate analysis and databases analysis,inflammatory-related potential biomarkers were screened and identified to analyze possible metabolic pathways.The reliability and biological significance of these biomarkers was verified by metabolic network analysis and correlation analysis with pharmacodynamic indicators.Results:A total of 16 potential biomarkers were screened and identified by multivariate analysis and metabolite databases,among which 13 species could be adjusted by RDN.The metabolism pathway analysis revealed that histidine metabolism,sphingolipid metabolism,and tyrosine metabolism were greatly disturbed.Their biomarkers involved urocanic acid,sphingosine,and norepinephrine,all of which showed a callback trend after RDN treatment.The three biomarkers had a certain correlation with some known inflammatory-related small molecules(histamine,arachidonic acid,Leukotriene B4,and PGE2)and pharmacodynamic indicators(IL-6,IL-1β,PGE2and TNF-a),which indicated that the selected biomarkers had certain reliability and biological significance.Conclusion:RDN has a good regulation of the metabolic disorder of endogenous components in carrageenan-induced inflammatory rats.And its anti-inflammatory mechanism is mainly related to the regulation of amino acid and lipid metabolism.This research method is conducive to the interpretation of the overall pharmacological mechanism of Chinese medicine.

A Network Pharmacology Study of Reduning Injection for the Treatment of Coronavirus Disease-19

Objective:The 2019 novel coronavirus(2019-nCoV)outbreak has escalated into a global pandemic.According to Chinese guidance for coronavirus disease-19(COVID-19):Prevention,control,diagnosis,and management,Reduning injection can effectively treat,the disease caused by the virus.To identify the active ingredients of Reduning injection and COVID-19 disease-related pathways,we conducted a network pharmacology study.Methods:The Traditional Chinese Medicine Systems Pharmacology database was used to screen the chemical constituents and potential targets of Reduning injection.The gene names were converted to the corresponding protein names using Uni Prot.Gene Cards and OMIM databases were used to select targets related to 2019-n Co V.Using Cytoscape 3.7.2 software platform and STRING database,we constructed drug-common target and target protein protein-protein interaction network diagrams.Rx643.6.2 software and Bioconductor biological information software package were used for Gene Ontology(GO)functional enrichment and KEGG pathway analyses.Results:In Reduning injection,a total of 33 effective chemical components were obtained that were involved in 151 signaling pathways,of which 44 targets were considered therapeutically relevant.Conclusion:Reduning injection can be potentially applied for the treatment of COVID-19 based on the results of our network pharmacology study.

Evaluation on Common Hand, Foot and Mouth Disease Treated by Integrative Traditional Chinese Medicine and Western Medicine

Objective To evaluate the efifcacy and safety of traditional Chinese medicine (TCM) combined with Western medicine in the treatment of patients with common hand, foot and mouth disease (HFMD) by conducting a prospective, controlled, and randomized trial. Methods A total of 452 patients with common HFMD were randomly assigned to receive Western medicine alone (n=220) or combined with TCM (Reduning or Xiyanping injections) (n=232). The primary outcome was the incidence rate of rash/herpes disappearance within 5 days, while secondary outcomes included the incidence rate for fever, cough, lethargy, agitation, and vomiting clearance within 5 days. Results The rash/herpes disappearance rate was 45.5% (100/220) in Western medicine therapy group, and 67.2% (156/232) in TCM and Western medicine combined therapy group, with significant difference (P<0.001). Moreover, TCM remarkably increased the incidence rate of secondary disappearance, which was 56.4%in Western medicine therapy group and 71.4%in TCM and Western medicine combined therapy group (P=0.001). No drug-related adverse events were observed. Conclusions It’s suggested that the integrative TCM and Western medicine therapy achieved a better therapeutic efficacy. TCM may become an important complementary therapy on relieving the symptoms of HFMD.

Effect of Integrated Chinese and Western Medicine Therapy on Severe Hand, Foot and Mouth Disease: A Prospective, Randomized, Controlled Trial

Objective: To evaluate the efficacy and safety of Chinese medicine(CM) plus Western medicine(WM) in the treatment of pediatric patients with severe hand, foot and mouth disease(HFMD) by conducting a prospective, controlled, and randomized trial. Methods: A total of 451 pediatric patients with severe HFMD were randomly assigned to receive WM therapy alone(224 cases, WM therapy group) or CM [Reduning Injection(热毒宁注射液) or Xiyanping Injection(喜炎平注射液)] plus WM therapy(227 cases, CM plus WM therapy group) for 7–10 days, according to a web-based randomization system. The primary outcome was fever clearance time, which was presented as temperature decreased half-life time. The secondary outcomes included the rate of rash/herpes disappearance within 120 h, as well as the rate for cough, runny nose, lethargy and weakness, agitation or irritability, and vomiting clearance within 120 h. The drug-related adverse events were also recorded. Results: The temperature decreased half-life time was 40.4 h in the WM therapy group, significantly longer than 27.2 h in the CM plus WM therapy group(P<0.01). Moreover, the rate for rash/herpes disappearance within 120 h was 43.6%(99/227) in the CM plus WM therapy group, significantly higher than 29.5%(66/224) in the WM therapy group(P<0.01). In addition, the rate for cough, lethargy and weakness, agitation or irritability disappearance within 120 h was 32.6%(74/227) in the CM plus WM therapy group, significantly higher than 19.2%(43/224) in the WM therapy group(P<0.01). No drug-related adverse events were observed during the course of the study. Conclusions: The combined CM and WM therapy achieved a better therapeutic efficacy in treating severe HFMD than the WM therapy alone. Reduning or Xiyanping Injections may become an important complementary therapy to WM for relieving the symptoms of severe HFMD.

Inhibition of Re Du Ning Injection on Enzyme Activities of Rat Liver Microsomes Using Cocktail Method

Objective Re Du Ning Injection （RDN）, a Chinese materia medica injection, is made from the extracts of LoniceraeJaponicae Flos, Gardeniae Fructus, and Artemisiae Annuae Herba. Since last decade, RDN has been widely used in China for the treatment of viral infection, fever, and inflammation. To assess the potential interacting of RDN with co-administered drugs, the inhibitory effects of RDN on the enzyme activities （CYP1A1, CYP1A2, CYP2C11, CYP2D1, and CYP3A1/2） of rat liver microsomes were investigated by a cocktail method. Methods A sensitive and specific LC-MS method capable of simultaneous quantification of five metabolites in rat liver microsomes was developed and validated. Then RDN （0.625%-1.0%） was incubated with rat liver microsomes and specific substrates. The enzyme activities were expressed as the formation rate of the specific metabolites of the substrates （pmol- mg. protein-1 . min-1）. Results RDN competitively inhibited the activities of CYP1A2 and CYP2C11, with inhibition constant （/~） values determined to be 0.18% and 0.63%, respectively. RDN exhibited the mixed inhibition on the activity of CYP2D1, with a K1 value of 0.15%. The activities of CYP1A1 and CYP3A1/2 were not markedly inhibited even by 1.0% RDN. Conclusion RDN could inhibit the rat enzyme activities of CYP1A2, 2Cll, and 2D1 in vitro with different inhibition modes, which is worthy of promoting safety and efficacy of RDN.

Regulation of RDN on Th1/ILC1 cell imbalance in HFMD patients caused by EV71 infection

Enterovirus 71(EV71)infection is more likely to cause hand,foot and mouth disease(HFMD)in children,which can lead to neurogenic complications and higher mortality.As a commonly used clinical medicine,Reduning injection(RDN)helps to shorten the symptoms of patients with HFMD and facilitate the early recovery of children.However,the regulatory mechanism of RDN on the HFMD immune system disorder caused by EV71 remains to be discussed.This study collected detailed treatment data of56 children with HFMD who entered the affiliated Children’s Hospital of Nanjing Medical University during 2019.Retrospective analysis of clinical data showed that the symptoms of the RDN treatment group were improved compared with the untreated group.To explore its mechanism,the relevant detection indicators were detected by flow cytometry,enzyme-linked immunosorbent assay and realtime quantitative PCR.It was found that the number and function of innate immune(ILCs)and adaptive immunity(Th1,Th2 and secreted cytokines)were reduced,suggesting that RDN plays a role by regulating cellular immunity.The in vitro differentiation inhibition test further confirmed that RDN affected Th1 differentiation by inhibiting the expression of transcription factors on the basis of Th1 cell differentiation in vitro.