Associations Of WT1 Gene And Other Factors With The Distribution Of TCM Syndromes In Acute Myeloid Leukemia

Objective:Using a retrospective study method,TCM syndrome types of AML patients were studied to explore the relationship between TCM syndrome types and patients'gender,TCM syndrome types and age,TCM syndrome types and WT1 gene level,TCM syndrome types and disease risk stratification,so as to provide a new reference direction for TCM prognosis of acute myeloid leukemia.Methods:According to the study requirements,144 patients with acute myeloid leukemia who met the inclusion criteria and were hospitalized in the hematology department of the First Teaching Hospital of Tianjin TCM University from September in 2017 to September in 2019were selected.All patients had received chemotherapy and were evaluated according to The Criteria for the Diagnosis and Efficacy of Hematologic Diseases and The Guidelines for Diagnosis and Treatment of Acute Myelogenous Leukemia?relapse/refractory?in China?2017?.The four examinations data of patients were collected to determine the TCM syndrome types,and the patients were divided into five syndrome types,namely,deficiency of qi and blood,deficiency of qi and Yin,excessive toxic heat,poison and blood stasis,deficiency of health qi and excess of pathogenic qi.In addition to the measured value of WT1gene?The WT1 gene was detected before the next course of chemotherapy after the myelosuppression period?,the data of bone marrow cell morphology,the result of chromosome examination and Mutations associated with AML were collected.Combined with the above findings,patients with AML were risk stratified.After the above data analyzed by SPSS 26.0,We respectively study the relationship between TCM syndrome types and patients basic information?sex,age?,the relationship between TCM syndrome types and WT1 gene expression level,the relationship between WT1 gene level and the disease risk stratification,the relationship between disease risk stratification and WT1 gene level,the relationship between subtypes of AML?M₁-M₇,not including the M₃?and WT1 gene expression.In this study,the real-time quantitative PCR detection of WT1 gene in all patients was determined by Beijing Heist Medical Laboratory,and the m RNA expression of WT1 gene in samples was detected by Taqman probe fluorescence quantitative PCR method.The expression level of WT1 is expressed as the ratio of WT1 copy number to ABL copy number multiplied by10000.Results:?1?All patients received chemotherapy,including 59 patients with remission,39 patients without remission,and 46 patients with relapse/refractory.Among the 144 patients,there were20 cases of deficiency of qi and blood?13 cases of remission,5 cases of no remission,2 cases of relapse/refractory?,23 cases of deficiency of qi and Yin?21 cases of remission,2 cases of no remission?,28 cases of excessive toxic heat?7 cases of no remission,21 cases of relapse/refractory?,8 cases of poison and blood stasis?2 cases of no remission,6 cases of relapse/refractory?,and 65 cases of deficiency of health qi and excess of pathogenic qi?25 cases of remission,23 cases of no remission,17 cases of relapse/refractory?.Among them,the number of patients with deficiency of health qi and excess of pathogenic qi syndrome is the largest,while the number of patients with only the deficiency syndrome?deficiency of qi and blood,deficiency of qi and Yin?or the excess syndrome?excessive toxic heat,poison and blood stasis?is relatively small.?2?Among the 144 AML patients,64 were male patients?44.4%?and 80 were female patients?55.6%?,The rate between male and female was 4:5.The gender of TCM syndrome types in each group was statistically analyzed,and the difference was not statistically significant?P>0.05?.Therefore,this study showed that the distribution of TCM syndrome types had no correlation with the gender of patients.?3?The 144 patients ranged in age from 18 to 79.The median age was 56±11 years with M±Q.All patients were divided into three groups:18-39 years old?20 cases?,40-59 years old?66 cases?and?60 years old?58 cases?.Then,the WT1 gene expression of patients in each age group was statistically analyzed,and the distribution of TCM syndromes in AML patients was found to be age-related?P<0.05?.Among patients aged 18-39 years old,the proportion of patients with deficiency of qi and Yin is the largest,and the proportion of deficiency of qi and blood syndrome is similar to that of deficiency of health qi and excess of pathogenic qi syndrome.Among patients aged 40-59 years old,the number of deficiency of health qi and excess of pathogenic qi syndrome was the largest,accounting for 50%of the total,followed by the excessive toxic heat?22.7%?.The proportion of deficiency of qi and blood syndrome and deficiency of qi and Yin syndrome was similar,and the number of cases of poison and blood stasis syndrome was the least.In patients?60 years old,the deficiency of health qi and excess of pathogenic qi syndrome proportion was the highest?44.8%?,followed by the excessive heat,and the other three cases were the similar.?4?The expression of WT1 gene in each age group was analyzed,and the difference was found to be statistically significant?P<0.05?.Therefore,there was a correlation between the expression level of WT1 gene in each age group.Pairwise comparison of each age group showed that the difference between the 18-39 group and the 40-59 group was statistically significant?P<0.05?.The median expression level of WT1 gene in patients aged 40-59 was higher than that in patients aged 18-39.The median expression level of patients aged 18-39years was higher than that of patients aged?60 years,and the difference between the two groups was statistically significant?P<0.05?.There was no significant difference between40-59 years old group and?60 years old group?P<0.05?.?5?In the relationship between AML subtypes and WT1 gene expression level,the distribution of WT1 expression level in AML subtypes was different after statistical analysis?P<0.05?.By comparison of AML subtypes,only M₁and M₅,M₂and M₅showed statistically significant differences?P<0.05?.The expression level of WT1 in patients with M₅was higher than that in M₁group and M₂group,and pairings of other groups showed no statistical significance in this study?P>0.05?.?6?The expression level of WT1 gene in each risk stratification group was calculated,and it was found that the expression level of WT1 gene was related to the risk stratification of disease?P<0.05?.The expression level of WT1 gene in the high-risk group was higher than that in the low-risk group?P<0.05?,and the expression level of WT1 in the medium-risk group was also higher than that in the low-risk group?P<0.05?.There was no significant difference in gene expression between the high-risk group and the medium-risk group?P>0.05?.This indicates that the expression level of WT1 gene is high in the high-risk group and the medium-risk group,while relatively low in the low-risk group.Since different risk stratification of AML indicates different prognosis,those with high expression level of WT1gene have poor prognosis,while those with low expression level have relatively good prognosis.?7?Among the relationship between the disease prognosis stratification of AML and TCM syndrome types,the proportion of low-risk patients with deficiency of qi and blood and deficiency of qi and Yin was the largest?85.0%and 82.6%,respectively?,and the number of patients in the middle-risk group and high-risk group was very small.The proportion of high-risk patients with excessive toxic heat or poison and blood stasis was the largest?60.7%and 62.5%,respectively?.In the deficiency of health qi and excess of pathogenic qi,the patients in the middle-risk group had the most cases,accounting for 40.0%.The test was statistically significant by Fisher's exact probability method?P<0.05?,suggesting that 5 TCM syndromes of AML patients were associated with disease risk stratification.?8?The m RNA expression levels of WT1 gene in 5 TCM syndrome groups were calculated,and the expression levels of WT1 gene in 5 groups were statistically different?P<0.05?,indicating that the distribution of TCM syndrome types was related to the expression level of WT1 gene.We found that the expression level of WT1 gene was the highest in patients with excessive toxic heat or poison and blood stasis,followed by deficiency of health qi and excess of pathogenic qi,while the expression level of WT1 gene was the lowest in patients with the syndrome of deficiency of qi and blood or deficiency of qi and Yin.Conclusion:The distribution of TCM syndromes of AML was not correlated with patient gender,but with patient age,AML subtypes,WT1 expression level and risk stratification of AML.The expression of WT1 gene was high in elderly patients,and the expression of this gene was higher in M₅patients than in M₁and M₂groups.WT1 gene expression was also correlated with risk stratification of AML.The disease risk stratification indicates different prognosis results,so the TCM syndrome type and WT1 expression level of AML also indicate the prognosis to a certain extent.The prognosis of the patients with the syndrome of excessive toxic heat and the syndrome of poison and blood stasis is poor.Patients with high levels of WT1 gene expression also had poor prognosis.This study has guiding significance to the prognosis judgment of the treatment of AML by TCM syndrome differentiation.