Study On The Treatment Of Advanced Stage Ovarian Cancer With Qi Deficiency And Blood Stasis By Regulating Tregs Related Snps In Yiqi Huoxue Jiedu Decoction

Ovarian cancer’s early diagnosis of the technology has not improved,because its early symptoms are not obvious and lack of specificity.And it is too late to found the patient’s problem.Although its standard treatment of chemotherapy and surgery has been exploring,but its effect has not yet achieved satisfactory results,and this kind of disease is recurrent.So ovarian cancer has been a major female health killer.Approximately 70%of patients were found to have advanced treatment for the late stage,which is currently the largest level of tumor cell reduction combined with platinum and paclitaxel in the first-line chemotherapy regimen.In recent decades,with advances in medical technology and the discovery and use of chemotherapy drugs,surgery and chemotherapy have made great progress,but the US epidemiology and database shows that nearly three decades of advanced ovarian cancer 5 years of survival The rate has not improved significantly.In recent years,Chinese medicine for the treatment of ovarian cancer patients have been concerned about.It not only can extend the time of patients without progression,but also can improve patient quality of life,reduce radiotherapy and chemotherapy side effects.The achievement of these advantages are recognized by individuals.After preoperative clinical study we found that:advanced ovarian cancer patients with Qi deficiency and blood stasis syndrome is the main clinical syndrome.So my mentor Professor Lu Wenping for the type of patients with self-designed Yiqihuoxuejiedu Decoction of the clinical syndrome differentiation treatment.This prescription is on basis of the "Li Chong Tang" which is created by modern Chinese medicine master Zhang Xichu for the treatment of gynecological tumor and it is also combined with our administrative office’s work experience of Sun Guizhi Professor,Piao Bingkui.Professor.Sun Guizhi and Piao Bingkui is the famous Chinese medicine expert of Guang’anmen Hospital of Chinese Academy of Traditional Chinese Medicine,and has unique opinions on the treatment of gynecological malignancies.Professor Sun said that ovarian cancer in Chinese medicine belongs to the abdominal mass,mostly due to qi stagnation,phlegm coagulation knot.When women menstrual or postpartum catch cold,they would caused by cold blood stagnation also due to emotional internal injuries.Qi stagnation of the course of time can make poisonous evil into cancer.The main drug composition contains fifteen kinds of drugs,for example:Astragalus,Yam,Atractylodes,Medlar,Ligustrum Lucidum,Curcuma Zedoary,Rhizoma Spargani,Pericarpium Citri Reticulatae Viride,Seaweed,Sweet Wormwood,Hedyotis Diffusa,Maris Pentadactyla,Scorpio and so on.Astragalus is the strongest strength of qi,it is the best kind of drug for qi in this prescription.Yam and Atractylodes can nourish the spleen for a kind of substance named qi.From the fundamental regulation of the human being’s qi generation,people who aim to cure the disease must be in the purpose of this.Qi is the leader of the blood,blood is the mother of qi.Doctors should pay attention to make up the body of the essence qi and blood,making them in harmony,so add Ligustrum Lucidum and other blood products.Curcuma Zedoary,Rhizoma Spargani,Pericarpium Citri Reticulatae Viride,Seaweed,Maris Pentadactyla,Scorpio,and other objects with blood circulation function.Strengthen Qi and focus on blood circulation at the same time.Artemisia Annua,Hedyotis Diffusa function is detoxification.To sum up,Li Chong Tang can make individuals stronger and reduce stasis,add insects take its advantages of deep illness,increase the power of blood circulation.Add some of the products of Hedyotis Diffusa,Artemisia Annua for the bad effect of fluid decrease.Tregs cells are a new perspective in the study of ovarian cancer by modern oncology.Modern research has found that the pathogenesis of tumors is more related to the immune system,and Tregs cells are an important part of the immune system.Immunotherapy is considered to be the most promising therapy to break through the existing treatment,but the mechanism of complex immunosuppressive molecules in tumor microenvironment and the lack of understanding of individual immune differences in ovarian cancer have made no significant progress in clinical practice.Recent studies have shown that regulation of T cells(Tregs)is a key immunosuppressive factor in immune immune escape and angiogenesis in complex immune networks of ovarian cancer.The polymorphism(SNPs)of related genes are closely related to the prognosis of ovarian cancer.The relationship between the mechanism of Tregs and ovarian cancer has a positive effect on the clinical development of ovarian cancer.CD24 is involved in cell adhesion and metastasis,it is confirmed to be platelet and endothelial cell adhesion receptors,leading to tumor cells in the blood transfer,and is considered a new carcinogenic factor,but also an important indicator of tumor diagnosis and prognosis.And related studies have shown that CD24-/-Treg cells can exert immunosuppressive function by secreting inhibitory cytokines(IL-10,TGF-p.IL-35).Through the clinical preliminary study we found that Qi and blood detoxification side can regulate the level of peripheral blood Tregs,enhance the immunity of patients with advanced ovarian cancer,reduce its immune suppression function,improve the clinical symptoms of patients,improve their quality of life.The preliminary experimental study also found that Yiqi Huoxue Jiedu combined with DC vaccine on the growth of transplanted mice have an inhibitory effect,the efficacy of immunosuppressive factors TGFβ1 and IL-10,and these two cytokines derived from Tregs.Therefore,we speculate that the side to extend the survival of advanced ovarian cancer may be related to the regulation of Tregs-related immunity.(SNPs)of Tregs-related genes related to the efficacy of syndrome differentiation and treatment,and the effect of this side on the expression of corresponding marker gene,the effect of prescription combination,the mechanism of intervention of traditional Chinese medicine and the mechanism of Chinese medicine intervention were discussed.Gene polymorphism,and provide the basis for the individual treatment of traditional Chinese medicine.Purpose:1 Screened with the benefits of qi and blood detoxification side effects related to SNPs.2 To explore the impact of Herb of mRNA expression corresponding to the difference between SNPs.Method:In this study,300 patients with advanced ovarian cancer were treated with syndrome of Qi deficiency and blood stasis.After taking yiqihuoxuejiedu tang six months,we take about 10-15 ml blood by using Illumina HumanExome-12v1-1.(TIANGEN TIANAMP Blood DNA Kit)was extracted from the blood of 300 patients with advanced ovarian cancer patients with Qi deficiency and blood stasis type.The samples were extracted from the blood of 300 patients with advanced ovarian cancer.GDNA was used to detect the concentration and purity of the whole genome.After hybridization,the nonspecific binding DNA was removed by washing,and the specific binding sites were left for single base extension.After staining,the cells were scanned with Illumina iScan Reader,The iScan system scan the original data into the Illumina official software GenomeStudio Genotyping module,for the use of the module for data analysis.Using GeneChipRPrimeView TM Human Gene Expression Array chip,mRNA was extracted from blood Qi and Blood advanced ovarian cancer patients 30 patients.(PFS),and the number of cancer antigen 125(CA125)records and quality of life after treatment evaluation,access to literature,advanced ovarian cancer patients with no progress in the survival of 16 months,The patients were divided into two groups,one group for the effective group of traditional Chinese medicine,the PFS>no progress in the median survival,the other group of invalid Chinese medicine group,the PFS ≤ no progress in the median survival.Through the comparison of SNPs between the effective and the ineffective SNPs,the SNPs were screened out,ie SNPs that could be sensitive to Chinese medicine.GO analysis:The SNP loci were significantly differentiated by the gene using the GO database to enrich the function.The GO functional class,Pathway analysis:The obtained gene was enriched by the KEGG database,and the biological pathway of the gene was significantly enriched.The mRNA expression of the effective and inactive groups was compared,and the differentially expressed genes were screened.And Pathway analysis,the difference between the GO function category and the biological pathway were obtained.The SNPs loci were used to identify the genes and differentially expressed genes.Result:1 There were 1773 SNPs which have significant differences.And there were 1033 SNPs on exons,970 SNPs for non-synonymous mutations.With GO enrichment analysis of differentially expressed genes,we found GO:0004955 and GO:0045087 had some relationship with immune system.GO:0006955 had some relationship with the body’s immune response and GO:0045087 had some relationship with congenital immune response.There were 18 genes enrichment to GO:0006955.They were FYB,RGS1,SUSD2,FCGRT,IL18RAP,CR2,IFI44L,CD96,C8A,CD1C,CXCR5,IL16,SEMA4D,PDCD1LG2,HLA-DMA,MICA,PKHD1L1 and HLA-DPA1.There were 24 genes enriched to GO:0045087.They were F18,TRAF3,CD180,EGF,NLRC5,C8A,NLRP4,IFI16,TLR3,FRS2,CD14,TAB2,FCN1,MAP1K1,GAB1,DTX4,VNN1,CR2,FOXO3,WIPF3,ITPR2,CD19,BPIFB3 and DEFB132.There were 14 major differences pathway in the KEGG enrichment analysis of the mutant gene,which mainly involved olfactory conduction,tyrosine metabolism,glycosylphosphatidylinositol(GPI),biosynthesis,fatty acid degradation,arginine and proline Amino acid metabolism,β-alanine metabolism,scopolamine,piperidine,pyridine alkaloid biosynthesis,pyruvate metabolism,long-term depression,flavonoid and flavonol biosynthesis,phenylalanine metabolism,PPAR signaling pathway,Base excision repair,taste conduction and other signal pathways.2 Through the analysis of mRNA differential expression,158 significant genes were obtained,including 58 up-regulated genes and 100 down-regulated genes.The GO analysis of the up-regulated genes was found to be related to immune-related GO 6 in the field of biological function,GO:00050277,GO:0050777,GO:0002227,which are associated with innate immune responses on the mucosa,immune responses,positive regulation of T cell immune response to tumor cells,GO:0006275,GO:0002842,GO:0002768,GO:0050776,Regulation,immune response regulation of cell surface receptor signaling pathway,immune response regulation of biological function;by down-regulation of the differential gene GO analysis found in the field of biological function and immune-related GO No.10,GO:0002566,GO GO:000453,GO:0045824,GO:0002286,which are associated with somatic cell mutation caused by immune receptor diversity,the immune system Positive regulation,immune response in the regulation of cytokines secretion,circulating immunoglobulin-mediated humoral immune response,immune response,helper T-cell Ⅰ-type immune response,congenital Activation reaction,innate immune response,a negative regulator of innate immune responses,activation of T cells related to immune responses.MRNA uptake gene uptake Passway enrichment obtained 8 related pathways,mainly related to asthma,malaria,hematopoietic cell lines,TGF-β signal transduction pathway,Toll-like receptor signal transduction pathway,phagosome,allograft rejection,tumor transcription disorder;down-regulated in the relevant passage 3,mainly related ABC transporters,lysosomes,phagosomes.3.According to the gene and expression profile of the GO analysis in the immune-related function on the intersection to know:with the benefits of qi and blood detoxification effects related to the possible immune function of the immune system is the response,the specific GO number is GO:0006955,Genes were enriched with 18 genes on the term,specifically:FYB,RGS1,SUSD2,FCGRT,IL18RAP,CR2,IFI44L,CD96,C8A,CD1C,CXCR5,IL16,SEMA4D,PDCD1LG2,HLA-MICA,PKHD1L1,HLA-DPA1,and the genes that were enriched in the term were CEACAM8,DEFA1,DEFA1,THBS1,CD24.4.The possible differences in the efficacy of traditional Chinese medicine intervention genes:CD40,AFF3,EIF2AK3,WNK1,FYB,FBXL13,PLXNC1,BCL11A,IFI16,GNG7.5.Yiqi Huoxue Jiedu Decoction may play an immunosuppressive function by down-regulating the expression of CD24 gene in patients with advanced ovarian cancer,but this process does not involve changes in SNPs.6.Yiqi Huoxue Jiedu Decoction can regulate Tregs cells by regulation of genes and related pathways,but are not significantly associated with SNPs and may be related to DNA demethylation or miRNAs.Conclusion:1 Associated with the efficacy of detoxification parties Yiqihuoxue SNPs locus 1773,these sites may be sensitive to traditional Chinese medicine SNPs.2 Yiqi Huoxue Jiedu Decoction The difference in marker gene may be:CD40,AFF3,EIF2AK3,WNK1,FYB,FBXL13,PLXNC1,BCL11A,IFI16,GNG7.3.Herb of Qi and Blood can be improved in patients with advanced ovarian cancer,immune function,resulting in difference in efficacy.4.Yiqi Huoxue Jiedu Decoction can regulate Tregs cells by regulation of genes and related pathways,but are not significantly associated with SNPs and may be related to DNA demethylation or miRNAs.