Study On The Decomposition Effect For Micromolecule Urotoxin By Intestinal Bacteria In Patients With Uremia Subjects

Objectives:To explore the transform of Intestinal flora and the decomposition effect to micromolecule urotoxin by Intestinal Bacteria in Patients with uremia.Methods:1.Experiment with sub-groups:uremic patients and normal subjects of the 48 cases,using random number table is divided into six subgroups.If the sub-group did not have more than one train a bacterial specimens from the group random fill.2.Experimental steps:Specimen processing:preparation of sterile specimen tube.Alcohol lamp flame 15cm range of subjects from fresh feces(30min～120min), said in the electronic scale of 3 specimens,each 1g,aseptic packaging in vitro 3.Sterile operating room will be three specimens of each test tube with sterile diluted into sterile saline 1:5,1:10,1:20 dilution and mixing different.Application of sample:biological safety cabinet(Clean Benches) using micro-pipette gun respectively mixing the above-mentioned coating liquid 2μl in blood agar.Incubation:The media buy in 37℃incubator to observe the results of a 24-hour incubation.Flora identification:observation of colony characteristics,staining, microscopy,and biochemical identification.Records:Counting the number of flora in agars.Ability to make use of toxin detection in urine:creatinine,said taking 15mg,uric acid 14mg,urea 40mg,slightly soluble in normal saline,the filter in 0.22μm microporous bacteria filter,then add 100ml of nutrient broth,shaken after the sub-installed in 1.5ml eppendorf tubes, each tube 1ml.Choose,respectively,normal and uremic patients with a bacterial colony Add eppendorf tube above,the mixing in the vortex oscillator into culture 37℃constant temperature incubator 24h,put up a control group,before and after training in the Beckman detector Blind Detection of media concentration in the urine toxin.The attention of the entire experimental process of asepsis.Results:(X±SD)1.flora analysis(×10⁸·g^-1)Compare of normal group(A) and Uremia group(B):Escherichia coli:A2.6±0.12,B 11.8±0.21,Escherichia coli increase in group B(P＜0.001).Enterococcus faecalis:A2.12±0.09,,B 3.46±1.01,Enterococcus faecalis increase in group B(P=0.007).Proteus vulgaris:A0.07±0.03,B 0.13±0.02,Proteus vulgaris increase in group B(P=0.001).Enterobacter cloacae:A0.14±0.03,B 0.24±0.05,Enterobacter cloacae increase in group B(P＜0.001).Citrobacter freundii:A 0.23±0.1,B 0.92±0.13,Citrobacter freundii increase in group B(P＜0.001).Bacillus subtilis:A0.12±0.03,B 0.32±0.05,Bacillus subtilis increase in group B(P＜0.001).Bacterial distribution..Group A Escherichia coli is more than Enterococcus faecalis(P＜0.001).Enterococcus faecalis is more than Citrobacter freundii(P＜0.001).Citrobacter freundii is more than Enterobacter cloacae(P=0.041).Enterobacter cloacae and Bacillus subtilis have no differences(P=0.152).Enterobacter cloacae and Bacillus subtilis are more than Proteus vulgaris(P＜0.001.P=0.005).Group B Escherichia coli is more than Enterococcus faecalis(P＜0.001) Enterococcus faecalis is more than Citrobacter freundii(P＜0.001) Citrobacter freundii is more than Bacillus subtilis(P＜0.001).Bacillus subtilis is more than Enterobacter cloacae(P=0.007).Enterobacter cloacae is more than Proteus vulgaris(P＜0.001).2.The decomposition effect for micromolecule urotoxin:1) Compare of control group(A),normal group(B) and Uremia group(C)Decompose creatinine(umol/L):Escherichia coli:A2.43±0.94,B 46.18±10.12,C 100.69±11.94.Group B and Group C can decompose creatinine(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Enterococcusfaecalis:A3.23±0.91,B23.54±3.66,C33.67±3.34.Group B and Group C can decompose creatinine(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Proteus vulgaris:A2.78±0.25,B 8.34±0.38,C 32.27±3.18.Group B and Group C can decompose creatinine(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Enterobacter cloacae:A 3.05±0.16,B 21.33±1.09,C34.52±1.43. Group B and Group C can decompose creatinine(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Citrobacter freundii:A2.34±0.15,B 4.52±0.11,C 12.17±0.15.Group B and Group C can decompose creatinine(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Bacillus subtilis:A1.03±0.28,B 2.28±0.15,C 8.8±0.75.Group B and Group C can decompose creatinine(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Decompose urea(mmol/L): Escherichiacoli:A0.02±0.01,B0.12±0.02,C2.74±0.07.Group B and Group C can decompose urea(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Enterococcus faecalis:A 0.04±0.02,B0.13±0.02,C 1.21±0.08.Group B and Group C can decompose urea(P＜0.001.P＜0.001),C is stronger than B(P＜0.001)Proteus vulgaris:A 0.03±0.01,B2.46±0.14,C9.66±0.15.Group B and Group C can decompose urea(P＜0.001.P＜0.001),C is stronger than B (P＜0.001).Enterobacter cloacae:A0.08±0.02,B 0.35±0.03,C1.38±0.26.Group B and Group C can decompose urea(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Citrobacter freundii:A0.06±0.01,B 0.27±0.04,C 0.95±0.07.Group B and Group C can decompose urea(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Bacillus subtilis:A0.06±0.01,B 1.09±0.14,C 2.14±0.1.Group B and Group C can decompose urea(P＜0.001.P＜0.001),C is stronger than B (P＜0.001).Decompose uric acid(umol/L):Escherichiacoli:A2.16±0.32,B21.85±1.26,C33.27±4.32.Group B and Group C can decompose uric acid(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Enterococcus faecalis:A 0.25±0.09,B0.73±0.18,C1.98±0.15.Group B and Group C can decompose uric acid(P＜0.001.P＜0.001),C is stronger than B(P＜0.001).Proteus vulgaris:A 0.66±0.12,B 0.76±0.16,C 1.44±0.1.Group C can decompose uric acid(P＜0.001),Group B can not decompose uric acid (P=0.244),C is stronger than B(P＜0.001)Enterobacter cloacae:A2.09±0.11,B3.43±0.27,C4.97±0.16.Group B and Group C can decompose uric acid(P＜0.001.P＜0.001),C is stronger than B(P＜0.001)Citrobacter freundii:A0.38±0.07,B0.43±0.09,C0.68±0.08.Group C can decompose uric acid(P＜0.001),Group B can not decompose uric acid(P=0.249),C is stronger than B(P＜0.001)Bacillus subtilis:A1.06±0.07,B 1.24±0.27,C 1.59±0.16.Group C can decompose uric acid(P＜0.001),Group B can not decompose uric acid(P=0.105),C is stronger than B(P=0.009)2) Compare the germs of uremia groupDecompose creatinine:Escherichia coli is stronger than Enterobacter cloacae(P＜0.001), Enterobacter cloacae and Enterococcus faecalis,Proteus vulgaris have no differences(P=0.528.P=0.089).Enterococcus faecalis and Proteus vulgaris have no differences(P=0.402).Proteus vulgaris is stronger than Citrobacter freundii(P＜0.001).Citrobacter freundii is stronger than Bacillus subtilis(P＜0.001).Decompose urea:Proteus vulgaris is stronger than Escherichia coli(P＜0.001), Escherichia coli is stronger than Bacillus subtilis(P＜0.001),Bacillus subtilis is stronger than Enterobacter cloacae(P＜0.001),Enterobacter cloacae and Enterococcus faecalis have no differences(P=0.111) Enterobacter cloacae,Enterococcus faecalis are stronger than Citrobacter freundii(P=0.002.P＜0.001)Decompose uric acid:Escherichia coli is stronger than Enterobacter cloacae(P＜0.001), Enterobacter cloacae is stronger than Enterococcus faecalis(P＜0.001), Enterococcus faecalis is stronger than Bacillus subtilis(P＜0.001), Bacillus subtilis is stronger than Proteus vulgar(P=0.048),Proteus vulgaris is stronger than Citrobacter freundii(P＜0.001).