| Objective:To analyze the clinical characteristics of children with ref ractory mycoplasma pneumoniae pneumonia(RMPP)with damp-heat syndrome,and to observe the clinical effect of clearing damp tongluo method on its trea tment.To study the effect of clearing damp tongluo recipe on the expressi on of IL-10,TNF-ɑ and IL-6 in RAW264.7 cells induced by mycoplasma pneumo niae(MP),and to explore the inflammatory intervention mechanism.Methods:1.Retrospective analysis method was adopted to collect 102 Hos pitalized children with RMPP(damp-heat syndrome)admitted to the affiliated hospital of shandong university of traditional Chinese medicine and shandong provincial hospital on January 1,2017 and December 31,2018.The clinical sympt oms,laboratory examinations,imaging characteristics and clinical efficacy were statistically analyzed.2.Preparation of serum containing clearing damp tongluo recipe:40 rats were randomly divided into blank control group,low,medium and high dose Chinese medicine group,and azithromycin group,8 each.The blank control group was treated with 0.9% normal saline 2m L/d.In the l ow-dose group,clearing damp tongluo decoction was used for 9.20g/(kg·d).In the medium dose group,clearing damp tongluo decoction was used as 18.40g/(kg·d).In the high-dose group,clearing damp tongluo decoction was used as36.80g/(kg·d).Azithromycin group was given azithromycin granule solution0.0645g/(kg·d)for 7 consecutive days.3.Establishment of mp-induced RAW264.7cell inflammation model:the model group and each drug administration group were respectively added with 100μL of MP bacterial solution,and the blank control group was added with 100μL DMEM complete culture solution,which was placed in the incubator for 2h.4.The intervention effect of clearing damp tongluo recipe on MP-induced RAW264.7 cells:The low,medium and high dose group and azithromycin group were given 10% serum containing 100μL,respectively,and the survival rate of RAW264.7 cells was detected by MTT method,and the expression levels of IL-10,TNF-a and IL-6 were detected by ELISA method.5.The minimum inhibitory concentration of single traditional Chinese medicine water decoction in clearing damp tongluo recipe was determined by microbroth dil ution method.Results:1.Clinical data characteristics:(1)The ratio of male to female in 102 RMPPP children was 1.17:1(P>0.05);The mean age was 6.75±3.06 years;The mean course was 15.39±5.26 d.(2)Chest CT:Unilateral lung involvement in 72cases(70.59%),bilateral lung involvement in 30 cases(29.41%);In the unil ateral image,50 cases(69.44%)had right lung involvement and 22 cases(30.56%)had left lung involvement.(3)Concomitant pathogen infection:single MP infect ion in 69 cases(67.65%),concomitant other pathogen infection in 33 cases(32.35%);There were only 19 cases of combined bacterial infection with MP(57.58%),10 cases of combined viral infection with MP(30.30%),and 4 cases of combined bacterial and viral infection with MP(12.12%).Gram-negative bacillus infection was the majority(60.87%)in patients with complicated bacterial infection,and RSV infection was the majority(28.57%)in patients with complicated virus infection.(4)The occurrence of complications in 70cases(68.62%),in the majority of patients with pulmonary atelectasis(71.43%).(5)Multiple linear regression analysis showed that CRP,LDH,d-dimer,mixed infection,and complications were statistically significant(P<0.05).2.Clinic al treatment comparison:(1)The overall curative effect of the disease and theoverall curative effect evaluation of TCM syndromes in the treatment group were superior to the control group(P<0.05).(2)The improvement of fever,co ugh,sputum cough,CRP and D-dimer in the treatment group was better than that in the control group(P<0.05).(3)The use of glucocorticoids and fiberoptic bronchoscopy in the control group was significantly higher than that in the treatment group(P<0.05).The average length of hospitalization was 13.16±3.47 d,there were significant differences between the two groups(P<0.05).3.MTT assay was used to detect the survival rate of MP-induced RAW264.7 cells:MP could significantly induce the proliferation and differentiation of RAW264.7 cells(P<0.05),and each drug administration group could inhibit the pr oliferation of mp-induced RAW264.7 cells(P<0.05).Azithromycin group had the best protection against cell proliferation(P>0.05 compared with the blank control group).4.The analysis of the expression of IL-10,IL-6 and the TNF-a of RAW264.7 cells MP-induced On the clearing damp tongluo method:The secretion of IL-10,TNF-a and IL-6 in RAW264.7 cells induced by MP was significantly increased(P<0.05).IL-10,IL-6 and the TNF-a were down-regulated in different degrees in each group compared with the model group(P<0.05),and the low,medium,and high dose groups showed a dose-dependent effect.5.MIC50 of Scut ellaria,Herba Artemisiae Capillariae,Coptis,Giant knotweed,Reed Rhizome,Ag astache rugosus,Acorus tatarinowii were 0.97,3.9,7.81,7.81,31.25,62.5 and62.5(mg/m L),respectively.MIC90 is 1.95,62.5,15.62,62.5,125,125,125(mg/m L).Conclusion:1.CRP,LDH,D-dimer,mixed infection and comorbidity were inde pendent influencing factors of RMPP.2.Clearing damp tongluo method has a positive effect on improving fever,cough,sputum and other symptoms,regula ting inflammatory reaction and microcirculation disorders.3.Clearing damp tongluo fang has a protective effect on the polarization and prolifer ation of mp-induced RAW264.7 cells in a concentration-dependent manner.4.Cleari ng damp tongluo fang down-regulated the high expression of IL-10,IL-6 and TNF-a in a dose-dependent manner.Clearing damp could inhibit the polarizationof RAW264.7 cells to both M1-type and M2-type,suggesting that it may play a regulatory role in maintaining the dynamic balance of M1/M2.5.In clearing damp tongluo prescription,Scutellaria,Herba Artemisiae Capillariae,Coptis,Giant knotweed,Reed Rhizome,Agastache rugosus,Acorus tatarinowii all have Inhibi tory effects on MP.Scutellaria has the best inhibitory effect on bacteria. |
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