Correlation Between Single Nucleotide Polymorphisms CYP3A4 Gene With Childhood ALL HD-MTX Use Caused Adverse

REACTIONS ABSTRACT CLINICAL STUDY OF ADVERSE CHILDHOOD ALL HIGH-DOSE METHOTREXATE THERAPYObjective: Discussion on the characteristics of HD-MTX regimen and a variety of adverse reactions, and explore the relationship between the various clinical manifestations in childhood ALL treatment.In order to provide useful ideas to be safe and effective treatment.Methods: Read carefully and analyzed 151 cases of childhood ALL cases available. Grouped according to the degree of risk stratification and adverse reactions. Compare the difference between clinical features and statistical analysis of the correlation.Results: In this study, patients were grouped according to CCLG2008 program risk standards for SR, IR, HR group. According to the degree of adverse reactions, divided into mild, moderate, and severe adverse reaction. And within each group and between different groups to compare data for statistical and comparative analysis.(1)To compare the incidence of bone marrow suppression: SR group and IR group the two groups, χ~2 = 12.716, p = 0.003 <0.05, statistically significantdifferences; SR group and HR between the two groups, χ~2 = 15.840, p =0.002 <0.05, statistically significant differences; IR group and HR between the two groups, χ~2 = 13.746, p = 0.003 <0.05, statistically significant differences.(2)To compare the incidence of liver injury: S group and IR group,χ~2 = 8.727,p = 0.004 <0.05,statistically significant differences; SR group and HR group, χ~2= 7.349, p = 0.003 < 0.05,statistically significant differences; IR group and HR group,χ~2 = 3.006,p= 0.240> 0.05, the difference was not statistically significant.( 3)Compare mucositis incidence:SR combination of IR group,χ~2 = 9.705,p = 0.003 <0.05, statistically significant differences; SR group and HR group, χ~2 = 19.371, p = 0.001 <0.05, the difference There were significant; IR group and HR group, χ~2 = 6.508, p = 0.021 <0.05,statistically significant differences.(4)To compare the incidence of nausea and vomiting of:SR group and IR group,χ~2 = 19.816,p = 0.001 <0.05,statistically significant differences; SR group and HR group, χ~2 = 8.260,p = 0.004 <0.05, statistically significant differences; IR group and HR group, χ~2 = 5.497, p = 0.026 <0.05, statistically significant differences.( 5) The occurrence of adverse reactions in 48 h plasma concentration<0.5μmol / L: bone marrow suppression in 85 cases 56.29% of the total sample, 82 cases of liver injury, accounting for 54.30%, mucositis in13 cases, accounting for 16.10%, nausea vomiting and other gastrointestinal symptoms in 8 cases, accounting for 5.30%. Adversereactions from delays occur.(6)the rate of adverse reactions occurring within SR, IR, HR group there are differences.Conclusion:(1)the case of bone marrow suppression:MTX dose is not the same in different incidence of bone marrow suppression, SR,IR, HR group rate were 17.31%, 57.83%, 100%, the degree of bone marrow suppression 5g/m~2 MTX than 2g/m~2 severe, so MTX-induced myelosuppression and dose-dependent; and in the same dose MTX, SR and IR group also found the incidence of different diseases because of individual differences in severity and MTX metabolism.( 2) Liver dysfunction: The same dose caused similar damage liver function, and increase the dose is increased incidence of liver injury.(3)In the case of mucositis: mucositis due to increased dose and increased incidence.(4)the case of nausea and vomiting: The incidence of dose due to increased and increased. But HR group compared with IR group, because the group of children with HR and more attention because of intense chemotherapy antiemetic drugs in advance, so the incidence of nausea and vomiting but lower than IR group. Above it shows that effective anti-emetic therapy may reduce the incidence of nausea and vomiting, is conducive to treatment.(5) 48 h plasma concentration <0.5μmol / L when, although the plasma concentration has been reduced to within safe limits, there are still some children began to adverse reactions. SR group, IR group, the incidence of HR between groups there is no difference, suggesting that unrelated toMTX dose. And all patients were normal renal function. It is concluded that adverse reactions are delayed due to metabolic MTX. Therefore,therapeutic drug monitoring period should be extended by a period should be further exploration work in practice.(6) Varying degrees of adverse reaction rate between S group and IR group and HR group, indicating the severity of adverse reactions with MTX dose-related, but also with individual differences, with MTX metabolic processes. Therefore, it is necessary to look for further causes of individual differences.CORRELATION BETWEEN SINGLE NUCLEOTIDE POLYMORPHISMS CYP3A4 GENE WITH CHILDHOOD ALL HD-MTX USE CAUSED ADVERSE REACTIONSObjective: Research on HD-MTX treatment in children with ALL the SNP CYP3A4 gene. And to explore the relationship between the SNP and an adverse reaction, which found that the SNP protective, and to explore the possible mechanism of influence of adverse reactions,improve useful help for clinical work.Methods:(1)To meet the requirements of collecting blood samples extracted genome DNA of white blood cells after separation.(2)Literature and find the point mutation CYP3A4*4、*5、*6、*18、*19and the rational design of primers. Obtained by PCR fragment containing each nucleotide of the SNP, sequencing, SNP statistical frequency of occurrence.(3)Statistical analysis of SNP loci with the relationship between adverse reactions, and to explore its effect on CYP3A4 enzyme metabolism and the effects of MTX.Results:(1)Compared with the normal population, this group of children with ALL CYP3A4*4* 5, *differences( p <0.05), which increases the incidence of CYP3A4*5、*6、*19,and CYP3A4*4、*6、*19 is reduced. CYP3A4*18 identical(p>0.05);(2)In children with mild adverse reactions, CYP3A4 *5 incidence of 0.66%, the incidence ofCYP3A4*18 2.65% occurrence rate, the difference was not statistically significant(χ~2 = 1.79, p> 0.05). Moderate adverse reaction incidence is1% difference. Occur in children with severe adverse reactions,CYP3A4*5 rate was 4.64%, no CYP3A4*18 appears;( 3) In the CYP3A4*5 genotype in children, 48 h MTX plasma concentration<0.5μmol/L, adverse reactions was 5.30%, no adverse reactions in children was 0.66%, statistically significant differences(χ~2 = 3.969, p<0.05). The CYP3A4*18 genotype in children, the difference was not statistically significant(χ~2 =6.72, p<0.05).Conclusion: These studies show that there is no difference CYP3A4*5 of CYP3A4*18 The incidence of mild adverse reactions in children,and more CYP3A4*5 children in the middle of severe adverse reactions occur, CYP3A4*5 genotype patients children exist MTX metabolism of delays. In cases of serious adverse reactions and delay the onset of,CYP3A4*5 expression more. CYP3A4*5 result CYP3A4 activity decrease, thus affecting the body’s metabolism of MTX. CYP3A4*18 is reversed, so that activity is increased.EXPERIMENTAL STUDY OF CYP3A4 ENZYMES IN RAT MODEL OF HD MTX TREATMENTObjective: Establishment of HD-MTX treatment Rats were observed to detect adverse events, get liver CYP3A4 enzyme expression, gain further CYP3A4 enzyme expression of the relationship between adverse reactions. And to explore the mechanisms that may exist in this process.Methods:(1)Construction of HD-MTX treatment model rats,observe the general status and mucosal damage,and liver function,blood indicators.(2)Adverse reactions grading.(3)To obtain the liver in rats using western blot method to detect CYP3A4 enzyme expression,investigate the mechanism of CYP3A4 enzyme having an impact on MTX metabolism..Results :( 1) Mild liver injury group and the moderate hepatic impairment group, the difference was not statistically significant( t=0.429, p>0.05), the expression of two liver enzyme CYP3A4 indifference. Mild liver injury group and severe liver injury group, the difference was statistically significant(t=-2.601,p<0.05). The expression of mild liver enzyme CYP3A4 injury group was higher than severe hepatic impairment group. Moderate hepatic impairment groups and severe liver injury group, the difference was statistically significant(t=-2.734,p<0.05), the expression of the enzyme CYP3A4 moderate hepaticimpairment group was higher than severe hepatic impairment group.(2)Group with mild bone marrow suppression and bone marrow suppression group difference was not statistically(t=1.004,p>0.05), two groups of CYP3A4 enzyme expression indifference. Mild bone marrow suppression and severe bone marrow suppression group group, the difference was statistically significant( t=-2.573, p<0.05), the expression of mild myelosuppression group CYP3A4 enzyme is higher than the group with severe myelosuppression. Moderate and severe bone marrow suppression group myelosuppression group difference was statistically significant( t=6.170, p<0.05), moderate myelosuppression CYP3A4 enzyme expression group was higher than the group with severe bone marrow suppression;(3)Mild and moderate mucosal damage mucosal damage and severe mucosal damage compared to all of the difference was not statistically significant(p>0.05),mucosal injury has nothing to do with the CYP3A4 enzyme expression.Conclusion : Injuries in rats caused HD-MTX, metabolism and CYP3A4 enzyme expression levels are correlated MTX,MTX CYP3A4 enzyme involved in the metabolism,decreased enzyme CYP3A4 may lead to slower metabolism and excretion of MTX delay.There is a difference in liver enzyme CYP3A4 content and bone marrow microenvironment CYP3A4 enzyme content and skin tissue CYP3A4 enzyme content, a variety of adverse reactions CYP3A4 enzyme expression correlation ofcause and there is a difference.( 2) group with mild bone marrow suppression and bone marrow suppression group difference was not statistically( t=1.004, p>0.05), two groups of CYP3A4 enzyme expression indifference. Mild bone marrow suppression and severe bone marrow suppression group group, the difference was statistically significant(t=-2.573,p<0.05), the expression of mild myelosuppression group CYP3A4 enzyme is higher than the group with severe myelosuppression. Moderate and severe bone marrow suppression group myelosuppression group difference was statistically significant(t= 6.170,p<0.05),moderate myelosuppression CYP3A4 enzyme expression group was higher than the group with severe bone marrow suppression...