Clinical Efficacy Of Xianglian No.2 Formula In The Treatment Of Tinea Pedis And Evaluation Of In Vitro Antifungal Activity

ObjectiveRandomized controlled clinical trials was performed.The treatment plan was used as the exposure variable(Chinese medicine XianglianNo.2formula group vs.0.25%amorolfine hydrochloride cream group)to evaluate the clinical efficacy of Chinese herbal medicine Xianglian No.2 formula in the treatment of tinea pedis.By in vitro experiment,different strains were treated with the optimized formula of Chinese medicine Xianglian No.2 formula,and the in vitro antifungal activity of the optimized formula of Chinese medicine XianglianNo.2formula was evaluated.At the same time,scanning electron microscope and transmission electron microscope were used to observe the microstructure of the fungus to further understand the effect of optimized formula of Chinese medicine XianglianNo.2formula on fungal.It is hoped that the anti-fungal effect of Xianglian No.2 formula can be objectively evaluated from different levels and different strengths of evidence.MethodsIn terms of randomized controlled clinical trials,this study randomly divided patients into treated groups and control groups.The treated group was a traditional Chinese medicine Xianglian No.2 formulacream group(n=33);the control group was a western medicine 0.25%amorolfine hydrochloride cream group(n=33).The researcher matched thetreated group and the control group at a ratio of 1:1.We employee a independent data-administrators to supervise and perform randomization.Randomization uses block randomize with randomized block size to achieve proportional randomization.The primary outcome is whether patient is cured(Y=1 is not cured;Y=0 is cured).The secondary outcomes include①Total scores for clinical symptoms and signs evaluation,the scores are mainly based on the evaluation of patients’ clinical symptoms and signs.Score;②Whether the fungus is cleared(Y=0 is cleared,Y=1 is not cleared);③Life scale score(dermatological quality of life scale;it is used as a continuous variable,respectively,before treatment,treatment After the 4th week,the 6th week after the treatment,and the 12th week after the treatment,evaluation will be carried out;④Drug safety assessment(the changes in liver function,blood routine index,and renal function before the medication,4,6,and 12 weeks after the medication).This study was approved by the ethics committee of the hospital when the research protocol was formulated.The investigator gave the subjects enough time to read and understand the informed consent statement,and signed the signature and date of consent.For the main outcome variables,The researchers used the chi-squared test and corrected with Pearson’s Chi-squared test with Yates’ continuity correction.In the study,the symptom and sign scores,blood routine,liver and kidney function,and quality of life scores are all repeated measurement data.Therefore,the generalized additive mixed model were employed for analysis.For in vitro experiments,Candida albicans(Candida albicans,×10 strains),Trichophyton rubrum(×10 strains),and Trichophyton interdigitale(Trichophyton interdigitale,×9 strains)were used as test strains,Taking the optimized prescription of Chinese medicine Xianglian No.2 formulaas the tested drug,observe the antifungal activities of the optimized prescription of Chinese medicine Xianglian No.2 formula against Candida albicans,Trichophyton rubrum,and Trichophyton interdigita.Observation of fungal microstructure changes under transmission electron microscope and scanning electron microscope at hour and 48 hours.ResultsClinical research results:No significant statistical difference was observed in the baseline data of the treatment group and the control group.This means that randomization eliminates the interference of confounding factors on the outcome,and the two groups are comparable.At 6 weeks after treatment of 33 patients in the Xianglian No.2 formula cream group,30 patients were cured and 3 patients were not cured.At 6 weeks after treatment,33 patients in the control group,21 patients were cured and 12 patients were not cured.The recovery rate of patients in the Xianglian No.2 formula cream group was higher than that of the control group,and the difference was statistically significant(P=0.008).The generalized additive mixed model is used to compare the change trend of the symptom score of different groups over time.It is found that at different time points(1-6 weeks after treatment),the symptom score of the treatment group decreases more than the control group,and the decline of the two groups The difference in chronological order was-0.95,-0.73,-0.58,-1.03,-0.66.The clinical interpretation is that at each time point,the symptom score of the treatment group decreased by 0.95,0.73,0.66 more than the control group points,1.03 points,0.58 points.The comparison of fungal clearance rate showed that 4 weeks(60.61%vs 87.88%)and 6 weeks(66.67%vs 90.91%)after treatment,the fungal clearance rate of tinea pedis patients in the control group was significantly lower than that in the treatment group,and there were statistical differences(P4week=0.016,P6 week=0.035).The results of the Dermatological Quality of Life Scale show that the two groups of drugs can improve the quality of life scores of patients,but the two have the same improvement in the quality of life scores.There is no evidence that there is a difference between the treatment group and the control group.The drug safety evaluation showed that at different time points after treatment,there was no statistical difference in hematology,liver and kidney function changes between the treatment group and the control group.In vitro experimental results:Xianglian No.2 formula has inhibitory effect on 10 strains of Candida albicans,and its minimum inhibitory concentration(MIC)is 15.625～62.5mg/ml;Xianglian No.2 formula has good effect on 10 strains of Trichophyton rubrum The minimum inhibitory concentration(MIC)is 7.8125～>250mg/ml;Xianglian No.2 formulahas good in vitro antibacterial activity against 9 strains of Trichophyton interdigitalis,and its minimum inhibitory concentration(MIC)is 15.625～31.25mg/ml.The results of electron microscopy at 24 and 48 hours after the medication showed that the hyphae of Trichophyton rubrum and interdigital fungus strains were twisted,folded,and surface rough,with different thicknesses,while Candida albicans ruptured and shattered.However,the above-mentioned fungal hyphae without drug action are smooth,uniform,and Candida albicans have complete morphology.The results of transmission electron microscopy at 24 and 48 hours after the treatment of the Xianglian No.2 formula showed that the cell wall structure integrity of the bacteria after the treatment of the drug was destroyed,the morphology under the cell wall microscope was obviously swollen,showing a loose appearance,and some severely damaged fungal cells The rupture and dissolution of the cell wall can be observed,and the cell membrane of the fungal cell is obviously swollen,showing a peeling appearance.Part of the cell membrane and cell nucleus and cytoplasm showed obvious pathological changes(fracture and dissolution of cell membrane,nucleus and cytoplasm,structural cavitation).In the negative control group,the complete cell wall structure,cell membrane integrity,cytoplasm(mitochondria,endoplasmic reticulum,glycogen granules,lysosomes,concentric membrane system,etc.)and cell nuclei were basically normal.And the longer the action time of the drug,the more obvious the damage of bacterial cells.ConclusionXianglian No.2 formula can significantly reduce the symptom score of patients with tinea pedis,and the effect is better than western medicine amorolfine hydrochloride cream.In addition,Xianglian No.2 formula showed better antibacterial activity in vitro.