The Study Of Mechanism And Therapeutic Effects Of The Integration Of Typhoid Fever And Blast Prescription For Viral Myocarditis

Background and aimAvian influenza virus and influenza virus have something in common:they are virus that invade the human body through respiratory tract to cause respiratory infection and general infection.Faced with the common challenge that more and more cases of respiratory virus infection,increasing severity of contagion, more seriousness of illness,it is essential for medical workers in China and western countries to find effective preventive and treatment methods to reduce the harm cause by respiratory virus infection.Viral myocarditis is an inflammatory disease,in which the heart wasinfected by virus.A lot of virus can cause Myocarditis,the research shows that influenza virus is the most common cause.But it's mechanism has not clearly understood yet,and lack of dependable therapeutical methods.Finding drugs from Chinese traditional medicine is new way for viral myocarditis therapy.There are certain advantages for Traditional Chinese Medicine(TCM)to prevent and treat this viral disease.Based on the characteristics of influenza virus and it's past experience of the respiratory viral infectious disease treatment,Professor Deng Tie-tao gained the clinically notable effects of treating influenza viruses in its early and middle stage by using the integration of typhoid fever and blast prescription.The anti-virus activity of typhoid fever and blast prescription has been found by many researchs.Our prevenient primary researchs showed that:the traditional Chinese medicine not only has a direct anti-virus activity in vitro,but also shows therapeutical effect on murine myocarditis in vivo.As a Avian influenza virus(AIV)model established in a laboratory has so many safety issues to be considered.The existing safety standard in our laboratory can not allowed us to conduct a real Avian influenza virus study,Influenza A virus(H1N1)is the only animal infectious respiratory virus in the Orthomyxoviridae family where Avian influenza virus also belongs to.Even though H1N1causes an acute respiratory infectious disease among human,it is still relatively safe to the research staff and environment under strict experimentalconditions.Therefore,it is our consideration of choosing FM1 as a AIV substitute to build a respiratory coronavirus infection like model. Based on what have been mentioned above,the main aims of this research are to establish the substituted Myocarditis Caused by Avian influenza virus model:and by using this model to evaluate the pharmacological action of the integration of typhoid fever and blast prescription and to some extent understand its mechanism of treating influenza virus infection.As a result, a further the integration of typhoid fever and blast prescription use in treating influenza virus infection may be developed.Aim:To observe the therapeutic effects of the typhoid fever and blast prescription extract on the Balb/c mice myocarditis caused by influenza A virus FM1 and to explore the mechanism of the effect.On the base of the models, to study and research mechanism and therapeutic effects of the integration of typhoid fever and blast prescription by Avian influenza virus,and to construct the base for study the integration of typhoid fever and blast prescription upon myocarditis caused by Avian influenza virus.Experimental partMethodsBalb/c mice were infected with the influenza A virus FMl by nasal route and batch sacrificedlate,on the base of the models,following experiment were carried. PartⅠThe treponin-cTnI were detected by enzyme-labeled immunosorbent assay (ELISA),the heart histopathologic(macro-and ultramicro)were observed by the light microscope and electron microscope.PartⅡMice were randomly divided into 6 groups,We cured the myocarditis of mice by intragastric administration for 14 days,and observed the body of weigh of mice,appetite,the symptom and the death of mice.We detected the rite between the heart mass and weigh of the body and macro-histopathologic and ultramicro changes of hearts.PartⅢThe NO and TNF-αin blood serum were examed by ELISA method to explore the effect of induction of NO and TNF-αby the four of drugs.PartⅣThe FM1 mRNA and TNF-αmRNA in heart were examed by FQ-PCR method to explore the effect of antivirus and induction of TNF-αby the four of drugs.ResultsPartⅠThe treponin-cTnI were detected in boold-serum were to be heightened during 3th-14th day after infected(P＜0.01).The heart histopathologic charge were inflammatory cell infiltration,dropsy and necrosis of heart cell.PartⅡThe four kinds of drugs can increase the body weight of myocarditis mice,appetite and the rate of survival;In increasing the rate of survival, the difference of the four groups is not significant.In increasing the body weight,all of the traditional Chinese medicine groups were higher than the model group in the 3th,5th,7th,10th and 14th days,the diffence was significant (P＜0.05;P＜0.01),virus azole group were higher than the model group in the 7th,10th and 14th days,the diffence was significant(P＜0.05;P＜0.01),the High dose group was higher than virus azole group and the low dose group,the diffence was significant(P＜0.05).there are dose-effect relationship of the integration of typhoid fever and blast prescription.The macrohistopathologic and ultramicro changes of mice hearts of the four drugs were better than the model group too,but the effect of the the High dose group and the the middle dose group was better than others.Precautionary effects were found of the High dose group on FM1 inffected of the Balb/c myocarditis mice(P＜0.05).PartⅢThe content of NO and TNF-αin blood serum of thefour groups were lower than the model group,the diffrence was significant(P＜0.05,P＜0.01),the content of NO in blood serum of the high group,the middle group and virus azole group were lower than the low group,the diffence was significant(P＜0.01); the content of TNF-αin blood serum of the high group was lower than other groups,the diffence was significant(P＜0.05).significant in the 7th and the 10th.The four kinds of drugs can inhibit replication of FM1 in heart.PartⅣThe mRNA expression of FM1 and TNF-αin the heart of three drugs groups of the traditional Chinese medicine were lower than the model group,the diffence was significant(P＜0.01).ConclusionPartⅠBalb/c mice were infected with the influenza A virus FM1 by nasal route and the cTnI were detected by enzyme-labeled immunosorbent assay(ELISA), the heart histopathologic(macro-and ultramicro)were observed by the light microscope and electron microscope,it is a easy test method to judge the myocarditis caused by Influenza A Virus.PartⅡThe four kinds of drugs can increase the body weight of myocarditis mice,appetite and the rate of survival.The macro-histopathologic and ultramicro changes of mice hearts of the four drugs were better than the model group,the effect of the the High dose group and the the middle dose group was better than others.Precautionary effects were found of the H group on FM1 inffected of the Balb/c myocarditis micePartⅢThe content of NO and TNF-αin blood serum of the four groups were lower than the model group.PartⅣThe mRNA expression of FM1 and TNF-αin the heart of three drugs groups of the traditional Chinese medicine were lower than the model group. Precautionary effects were found of the High dose group.Clincal partObjectiveTo investigate the clinical characteristics and differentiation of TCM syndromes of viral myocarditis,and to explore effective TCM regulations of diagnosis and therapy.Methods(1)Grade the commonly encountered symptoms and physical signs of viral myocarditis. (2)Fill out "pediatric viral myocarditis TCM syndromes questionnaire" in detailed,and classify the cases according to different TCM syndromes.(3)Analyze the relationship between age,course and TCM syndromes with statistical method,and study the distributing state of TCM symptomsand physical signs.(4)On the basis of TCM syndromes investigation,establish effevtive TCM regulations of virus myocarditis.Results(1)The chief symptoms of viral myocarditis are hypodynamia(54.46%),chest distress(49.11%),palpitation(29.46%),dizziness(25.00%),cacochroia (24.11%),pectoralgia(21.43%),Jie Dai Mai(13.40%),hidrosis(9.82%):the secondary symptoms are red throat(75.90%),texture of tonguea(88.39%),bnormal tongue(72.32%).fever(25.89%),cough(22.33%),abnormal stoo1(19.64%), nausea(9.82%),myalgia(8.04%).(2)The commonly encountered TCM syndromes of viral myocarditis are Xie Fan Xin Fei(68.75%),Qi Yin Bu Zu(24.11),Xin Mai Yu Zu(7.14%).(3)Xie Fan Xin Fei is the most commonly encountered TCM syndrome during acute stage,the differentiation of TCM syndromes distribution between acute stage and chronic stage has high statistical significance(P＜0.01).(4)The disease incidence of Xie Fan Xin Fei declined with increase of course, the disease incidence of Qi Yin Bu Zu and Xin Mai Yu Zu ascended with increase of course.The tendency of TCM syndromes incidence has high statistical significance(P＜0.01).ConclusionThe commonly encountered TCM syndromes of mild and moderate pediatric viral myocarditis can he summarized as:Xie Fan Xin Fei,Qi Yin Bu Zu,Xin Mai Yu Zu.hypodynamia,cacochroia,chestdistress,pectoralgia,fever,redthroat, nausea,tongue coat,pulse(fingerprint)and course have better discrimination. the discriminant function can discriminate TCM syndromes correctly,principle and method of treatment in TCM are the integration of typhoid fever and blast, expel evils and nourish Zheng Qi during initial stage;replenish Zheng Qi and eliminate residual evils during anaphase. There are certain peculiarities and new ideas in this study.Firstly,it was the model established by virus infection in vivo.This model has not been mentioned in the pharmacological literatures of China before.This myocarditis caused by influenza A virus model.which was established by choosing FM1 as a substitute of Avian influenza Virus would fill up the blank in this area.It would provide another channel for screening an effective Chinese or Westem medical drug.To some certain extent,this model also matched the TCM syndrome of respiratory virus infection.Therefore it has an important value for evaluating an anti-viral drug,especially a Chinese medicine compound with the functions of removing heat and damp acrid-warm herbs relieving superficies and anti-virus,although the respiratory coronavirus infection model was established by a none Avian influenza Virus.infection in this study,there was still a corelation between the two types of virus.This model have the following advantages over the others:easy establishment,good reproduc -ibility safety and no special laboratory need,short experimental period, economy,and limit threat of causing biological disaster ease of widely use, and several models establishment at one time.All these will be propitious for studying the anti-influenza Virus drugs especially for an initial screening of an anti-influenza Virus drug.Secondly,there are new ideas in this study. To observe the therapeutic effects of the typhoid fever and blast prescription extract on the myocarditis caused by influenza A virus FM1 and to explore the mechanism of the effect.On the base of the models,to study and research mechanism and therapeutic effects of the integration of typhoid fever and blast prescription by Avian influenza virus,and to construct the base for study the integration of typhoid fever and blast prescription upon myocarditis caused by Avian influenza virus.