| Objectives1.Clinical researchUnder the guidance of syndrome differentiation theory,using local differentiation theory to study the difference of clinical index,treatment and survival time of "damp-heat toxin" and"cold-heat toxin" syndrome in patients with non-small cell Lung cancer(NSCLC)with Malignant Pleural Effusion(MPE),providing an theoretical basis for TCM syndrome differentiation and exploring the relationship between TCM syndromes and the prognosis of patients.2.Experimental researchStudy the differential miRNA expression profiles between "damp-heat toxin" and "cold-heat toxin" syndrome patients,making target prediction of differential miRNA and analyzing the GO and KEGG pathway,find new biomarkers for MPE diagnosis,providing molecular biology objective basis of TCM syndrome differentiationMethods1.Clinical researchA retrospective study was conducted to collect the data of NSCLC patients with MPE who met the inclusion criteria and were admitted to DongFang hospital of Beijing University of Chinese Medicine from June 1,2013 to November 30,2019.Compare the difference of clinical index(including blood routine,blood biochemistry,blood coagulation function,serum tumor markers,pleural effusion routine,pleural effusion biochemical,pleural effusion tumor markers),treatment method(including combination of Chinese and western medicine therapy,Chinese medicine therapy,western medicine therapy,and alleviative therapy)and survival time(average survival time,1,6,12,24 months survival rate)between "damp-heat toxin" and "cold-heat toxin" syndrome patients after intercostal drainage.Then taking a further analysis to explore the treatment plan of patients with survival advantage.2.Experimental researchA total of 8 NSCLC patients(both "damp-heat toxin" and "cold-heat toxin" syndrome have 4 patients)were consecutively selected from a cohort of NSCLC patients with MPE.These cohort patients were diagnosed with NSCLC between January January 1,2019 and November 30,2019.These patients were enrolled from DongFang hospital.Collecting the pleural effusion samples while the patients taking intercostal drainage for the first time,extracting pleural effusion miRNA.Using High-throughput sequencing to explore the expression profiles of miRNA and filter the differential expression miRNA to predict the target gene.The predicted target genes were analyzed by GO gene functional enrichment and KEGG Pathway.Results1.Clinical research(1)General information and tumor-related informationA total of 241 cases were included,90(37.34%)patients were in "damp-heat toxin"syndrome group and 151(62.66%)were in "cold-heat toxin" syndrome group.There were no statistically significant differences in age,gender,smoking history,family history of tumor,and KPS score between the two groups(P>0.05).Both groups two groups had the highest proportion of right lung,lung adenocarcinoma,low differentiation,lung and lymphatic metastasis.101 patients were taken gene testing,the "damp-heat toxin" syndrome group had a high percentage of exon 19 deletion mutation,the "cold-heat toxin" syndrome group had a high percentage of exon 21 point mutations.There was no statistically significant difference in gene mutation sites between the two groups(P>0.05)(2)Laboratory values1)Blood routine:The RBC and HGB values of the two groups were lower than the standard value,MONO values were higher than the standard value.The WBC,LYM,PLT and MONO values of the "damp-heat toxin" syndrome group were higher than "cold-heat toxin" syndrome group,and the HGB values were lower than "cold-heat toxin" syndrome group There was no statistically significant difference in gene mutation sites between the two groups(P>0.05).2)Blood biochemistry:The TP,ALB and A/G values of the two groups were lower than the standard value.The ALT(P<0.05),AST(P<0.05),ALP(P>0.05),LDH(P>0.05)values of the "damp-heattoxin" syndrome group were higher than "cold-heat toxin" syndrome group,the IBIL(P>0.05),ADA(P>0.05)and GLU(P>0.05)values were lower than "cold-heat toxin"syndrome group.3)Coagulation function:The FIB and d-d values of the two groups were higher than the standard value.The FIB(P>0.05)and d-d(P>0.05)values of the "damp-heat toxin" syndrome group were higher than "cold-heat toxin" syndrome group.4)Blood tumor markers:The CEA and Cyfra21-1 values of the two groups were higher than the standard value.The CEA(P>0.05)and Cyfra21-1(P>0.05)values of the "damp-heat toxin" syndrome group were higher than "cold-heat toxin" syndrome group.5)pleural effusion test(routine,biochemical,tumor markers):The total number of cells(P<0.05),RBC(P<0.05),WBC(P<0.05),protein(P<0.05),ADA(P<0.05),LDH(P<0.05),NSE(P<0.05),and Cyfra21-1(P<0.05)of the "damp-heat toxin" syndrome group were higher than "cold-heat toxin" syndrome group,the GLU(P<0.05)and CEA(P>0.05)values were lower than "cold-heat toxin" syndrome group.(3)Treatment methodsThe highest proportion of the treatment methods of the two groups is combination of Chinese and western medicine(P>0.05).The proportion of perfusion with Chinese medicine injection of the "damp-heat toxin" syndrome group were higher than "cold-heat toxin"syndrome group(P<0.05).(4)Survival timeThe average survival time of "damp-heat toxin" syndrome group was 7.76 months,and 9.24 months in "cold-heat toxin" syndrome group(P>0.05).The 1,6,12 and 24 month survival rates were 73.33%,38.89%,13.33%and 3.33%in "damp-heat toxin" syndrome group,and 80.13%,41.72%,20.53%and 2.65%in "cold-heat toxin" syndrome group.The average survival time of the two groups who were not treated with chest cavity perfusion with traditional Chinese medicine injection was 6.14 months in the "damp-heat toxin" sydrome group.The average survival time was 9.38 months in "cold-heat toxin" syndrome group(P<0.05).2.Experimental research(1)Analysis of miRNA expression differencesThere were 100 miRNAs differentially expressed between the two groups,among which 56 miRNAs were upregulated in the "damp-heat toxin" syndrome group compared with the"cold-heat toxin" syndrome group,mainly related to the miR-200 family,and 44 down-regulated miRNAs.(2)Analysis of GO functionThe results of GO function analysis of the upregulated expression miRNA target genes were mainly involved in angiogenesis.cell development,mRNA metabolism,signal transduction protein phosphorylation,regulation of ion transmembrane transport,extracellular structure,and vascular morphogenesis,etc.The down regulation expression miRNA target genes were mainly involved in negative regulation of cell development,signal transduction protein phosphorylation,the regulation of T cell activation,extracellular structure,ion transmembrane transport,positive control,white blood cell differentiation,cell protein localization of negative regulation of mitotic cell cycle,etc.(3)Analysis of KEGG pathwayThe results of KEGG pathway analysis of the upregulated expression miRNA target genes were mainly involved in chromosome and related proteins,proteoglycan,phospholipase D signaling pathway,cAMP signaling pathway and platelet activation,etc.The down regulation expression miRNA target genes were mainly involved in proteoglycan,phosphoinositol metabolism,phosphatidylinositol signaling system,sphingolipid signaling pathway,phospholipase D signaling pathway,cAMP signaling pathway,autophagy,etc.Conclusions1.Clinical researchIn this study,we found that the local syndrome differentiation of NSCLC patients with MPE was characterized by "damp-heat toxin" sydrome,and the pathological type was mainly lung adenocarcinoma.There was no significant correlation between the two groups with gender,age,pathological type,family tumor history,smoking history,KPS score,and gene mutation type.The total number of cells,RBC,WBC,GLU,protein,ADA,LDH,NSE,Cyfra21-1 values of pleural effusion were correlated with the local syndrome differentiation of NSCLC patients with MPE,which could be used as the objective basis for the local syndrome differentiation.In addition,the prognosis of "damp-heat toxin" syndrome group was worse than that of "cold-heat toxin" syndrome group.Combination of Chinese and western medicine therapy is an effective method to improve the survival of patients.Using the local syndrome differentiation as an guideline to infusion Chinese medicine injection also benefits the patients.2.Experimental researchCompared with the "cold-heat toxin" syndrome group,the upregulate miRNA of "damp-heat toxin" syndrome group is mostly related to the poor prognosis,while the down-regulated miRNA is mainly related to the progress of tumor inhibition and the occurrence of drug resistance.The GO analysis results suggested that the biological processes involved in the differential expression of miRNA in the two groups mainly included the roles of angiogenesis,the regulation of T cell activation,and leukocyte differentiation.KEGG analysis results suggested that the KEGG signaling pathway involved in the differential expression of miRNA in the two groups was mainly related to PI3K-akt. |
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