| Purpose:Using an fermentation model in vitro batch to simulate the human's intestine to study the glycolysis of various oligosaccharides by intestinal microflora in children with acute lymphoblastic leukemia(ALL)during chemotherapy.After fermentation,gas chromatography was used to detect the content of short-chain fatty acids of feces in different oligosaccharides.The relationship between intestinal flora disturbance and bacterial metabolism in acute lymphoblastic leukemia patients during different chemotherapy periods was explored.Its aim is to provide a clinical basis of the probiotics or prebiotics administered early to children with acute lymphoblastic leukemia during chemotherapy.Materials and Methods: In this study,thirty children are with newly diagnosed and untreated acute B-lymphocytic leukemia with a average age of 5.12±2.82 years old and a ratio of male to female is 2:1.The time of feces collection was the VDLP course of CCCG-2015-ALL.According to faecal collection points,there were divided into pre-chemotherapy group,chemotherapy group(myelosuppression: neutrophil <0.5x109/L)and post-chemotherapy group(bone marrow recovery: neutrophils >0.5x109/L and platelets >80x109/L).In addition,there were 16 children with the intervention of probiotics in chemotherapy,and 14 children with non-intervention of probiotics?The normal group was 12 children with normal physical examination in children's health section,including 6 boys and 6 girls and the average age was 1.79±0.96 years old?Collecting and treating their fecal specimens,and then inoculating in FOS(oligofuranose),MOS(oligomannose),INU(inulin),XYI(xylitol),and YCFA(without any carbohydrate)oligomerization medium?After 24 hours of fermentation in the sugar medium,measuring the amount of gas and determing the content of short-chain fatty acids(SCFAs)in different oligomerization medium?SPASS 22.0 was used for statistical analysis of the data.Results:1.Comparison of SCFAs in Unfermented Feces1.1 The content of SCFAs in three groups of un-fermented feces of children with ALL compared with normal group :The content of total acid,acetic acid,propionic acid and butyrate of the pre-chemotherapy group(n=30)was higher than that of the normal group,but there was no statistical difference(P>0.05).The content of propionic acid of the chemotherapy group(n=14)was significantly higher than that of the normal group(P<0.05).The connets of the total acid,acetic acid,and propionic acid of the post-chemotherapy group(n=14),were significantly higher than those of the normal group(P<0.05).1.2 The content of SCFAs in the unfermented feces of the group without probiotics intervention(n=14)compared in themselves:The contents of total acid,acetic acid,propionic acid,and butyric acid in group without probiotics intervention(n=14)decreased first and then increased in the pre-chemotherapy group,the chemotherapy group,and the post-chemotherapy group.The content of butyric acid was significantly higher in the pre-chemotherapy group and the post-chemotherapy group than in the chemotherapy group(P<0.05).There was no statistical difference between the two groups in comparing the the other kinds of short-chain fatty acids(P>0.05).1.3 The content of SCFAs in the unfermented feces of group with probiotics intervention(n=16)compared in themselves:In the group with probiotics intervention(n=16),the contents of total acid,acetic acid,and propionic acid showed a rising trend in the pr-echemotherapy group,the chemotherapy group,and the post-chemotherapy group,while the content of butyric acid decreased first and then increased in the three groups.The content of all kinds of short-chain fatty acids in the three groups was no significant difference(P>0.05).1.4 Comparison of changes in SCFAs of unfermented feces in group(n=16)with probiotics intervention and froup without probiotic-untreated(n=14):In the group with probiotic intervention(n=16),the increase in total acid,acetic acid,and butyrate from before chemotherapy to myelosuppression(the contents of SCFAs in the chemotherapy group minus the pre-chemotherapy group)was significantly higher than that in the group without probiotic intervention(n=14)(P<0.05);There was no statistical difference in the content of short-chain fatty acids(SCFAs)from myelosuppressive phase to bone marrow recovery phase,and the same of period of throughout chemotherapy(P>0.05).2.Comparison of SCFAs and Gas Production after Fecal Fermentation2.1 The Content of SCFAs of Three Groups in Fermented Feces of Children with ALL Compared with Normal GroupSCFAs: Compared with the normal group,in the FOS medium,the contents of total acid acetic acid and propionic acid of the pre-chemotherapy group were significantly lower than that of the normal group(P<0.05).In the INU medium,the contents of total acid and acetic acid of the pre-chemotherapy group were significantly lower than the normal group(P<0.05).In the YCFA medium,the content of acetic acid of the chemotherapy group was significantly higher than that of the normal group(P<0.05).In the XYI medium,the content of totel acid of the chemotherapy group was significantly lower than that of the normal group(P<0.05)and the butyric acid content was significantly higher than that of the normal group(P<0.05).In YCFA medium,the content of acetic acid of post-chemotherapy group was significantly higher than the normal group(P<0.05).Gas production: The gas ofall the medium of the pre-chemotherapy group was lower than those of the normal group.The gas of the FOS,MOS,and INU medium of pre-chemotherapy group was significantly lower than those of the normal group(P<0.05).The gas ofall the medium(except FOS)of the chemotherapy group was lower than that of the control group.Statistical analysis showed that only in the MOS medium,the gas was significantly lower than the normal children group(P<0.05).There was no statistical difference between the post-chemotherapy group and the normal control group in the valuesof gas in all the media(P>0.05).2.2 The Content of SCFAs and Gas in the Fermented Feces of the Group without Probiotics Intervention(n=14)Compared in ThemselvesSCFAs: The content of butyric acid in INU medium was significantly higher of the chemotherapy group than of the pre-chemotherapy group(P<0.05).The content of propionic acid in XYI medium was significantly lower of the chemotherapy group than of the pre-chemotherapy group(P<0.05).There was no statistically significant difference in other kind of SCFAs between the every two groups(P>0.05).Gas production: The gas in FOS,MOS,and INU medium of the pre-chemotherapy group,the chemotherapy group,and the post-chemotherapy group showed an upward trend,while the gas in YCFA and XYI medium showed an increase and then a decrease trend in the three groups.The gas in FOS and INU was significantly higher than that pre-chemotherapy group(P<0.05)and the post-chemotherapy group was significantly higher than the pre-chemotherapy group(P<0.05).The gas in MOS medium of post-chemotherapy was significantly higher than that of pre-chemotherapy group and chemotherapy group(P<0.05).2.3 The Content of SCFAs and Gas in the Fermented Feces of the Group with Probiotics Intervention(n=14)Compared in ThemselvesSCFAs: The conents of total acid,acetic acid,propionic acid,and butyric acid contents in the YCFA,FOS,MOS,and XYI of the group with probiotic intervention showed an upward trend in the pre-chemotherapy group,the chemotherapy group,and the post-chemotherapy group.The content of acetic acid,in MOS medium,the content of total acid and acetic acid in INU medium,of the chemotherapy group were significantly higher than those of the prechemotherapy group(P<0.05).The contents of total acid in YCFA medium,the contents of total acid and acetic acid in MOS medium of post-chemotherapy group significantly higher than those of pre-chemotherapy group(P<0.05).Gas production: The gas in FOS,MOS,and INU of the chemotherapy group was significantly higher than the pre-chemotherapy group(P<0.05).The gas in MOS medium of the post-chemotherapy group was significantly greater than that of the pre-chemotherapy group(P<0.05).2.4 Comparison of Changes in SCFAs and Gas Production of Fermented Feces in Group(n=16)with Probiotics Intervention and Group without Probiotic The Content of SCFAs and Gas in the Fermented Feces of the Group without Probiotics Intervention(n=14)SCFAs: The increase of total acid and propionic acid in pre-chemotherapy in the XYI medium and increase of propionic acid in pre-chemotherapy in the YCFA medium of group with probiotic intervention from the pre-chemotherapy to the myelosuppression(the contents of SCFAs of the chemotherapy group minus that of the pre-chemotherapy group)was significantly higher than the group without probiotics intervention(P<0.05).The increase of the total acid and propionic acid of the group with probiotics intervention in YCFA medium and XYI medium(the contents of SCFAs of the post-chemotherapy group minus that of the pre-chemotherapy group)were significantly higher than the group without probiotics intervention(P<0.05).Gas production:The comparise of the increase of the Gas production in any period between the two grouphasno statistical difference.3.The content of butyric acid in MOS medium was higher than in other mediums(except XYI in chemotherapy group).The content of butyrate in MOS medium of pre-chemotherapy group was significantly higher than that in FOS medium and INU medium(P<0.05).Conclusion:1.The content of SCFAs was higher in the feces of ALL children in pre-chemotherapy group than in normal children.After the fermentation,the content of SCFAs in the feces of ALL children in pre-chemotherapy group was lower than that in normal children,suggesting that acute lymphoblastic leukemia can disturb the intestinal flora.2.The content of short-chain fatty acids(SCFAs)in feces of ALL children decreased first and then increased with chemotherapy.After fermentation,the content of SCFAs in all medium increased with the progress of chemotherapy,suggesting that chemotherapy can effect the intestinal flora.3.There were more SCFAs in the unfermented feces of the probiotic intervention group than in the untreated group.After the fermentation,the increase of the content of SCFAs of the group with probiotic intervention was higher in all culture medium than that of the group of the group without probiotic intervention.It suggests that probiotics which were oraled can survive in the intestine and may promote the growth of certain beneficial bacteria.4.Using MOS(oligomannose)as a substrate for fermentation can produce more butyric acid that is beneficial to humans.5.The determination of SCFAs and gas content in feces in oligosaccharide in vitro fermentation models may be helpful in providing early clinical evidence of oraling probiotics or prebiotics in children with acute lymphoblastic leukemia. |
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