Preliminary Investigation Of Research Intestinal Microflora In Patients With Hepatic Cirrhosis By FISH And LCSM

Objective:To examine the difference of fecal Bifidobacterium(probel:Bifl64 5'-CATCCGGCATTACCACCC-3',Fluorescence:TAMRA),Bacteroides(probe2:Bac303 5'-CCAATGTGGGGGACCTT-3,Fluorescence:HEX),Eubacterium rectale-Clostridium (probe3:Lowgc2p 5`-GGGACGTTGTTTCTGAGT-3,Fluorescence:TAMRA), Escherichia coli (probe4:Ecoli 5'-GCAAAGGTATTAACTTTACTCCC-3,Fluorescence: HEX), Enterococcus(probe5:Enc 5`-CCCTCTGATGGGTAGGTT-3,Fluorescence: FAX), Clostridium difficile (probe6:EREC482 5'-GCTTCTTAGTCAGGTACCG-3', Fluorescence:FAX)between patients with hepatic cirrhosis and healthy college students.Methods:29 samples of patients with hepatic cirrhosis were collected from departments of digestive of Zunyi hospital during the period January to October 2010,and compare with feceses of 13 cases in college stundents as a healthy control group, to detect pH value and bacterioturbidity, FISH(fluorescent in situ hybridization) on a slide, LSCM(laser scanning confocal microscopy)were performed for further analysis.T test and Rank correlation test methods were experiments for statistical analysis.Results:pH value of the feceses of the patients with hepatic cirrhosis was higher than healthy control group(P<0.05), but there was no significant difference between the hepatic cirrhosis group and healthy control group in bacterioturbidity(p>0.05).The fecal numbers of Bifidobacterium,Bacteroides,Eubacterium rectale-Clostridium,Escherichia coli, Enterococ-cus,Clostridium difficile of the patients with hepatic cirrhosis were significantly less than that of the controls group(P<0.01), the proportion of 6 kinds of bacterias was changed,mainly the number of obligate anaerobes as Bifidobacteria and Bacteroides were redued,and the number of E.coli, Eubacterium rectale-Clostridium, Enterococcus, ClostriDium difficile were increased(p<0.05).Between the number and the ratio of bacteria with cause and classifical severity of hepatic cirrhosis(Child-Pugh) was no significant correlation. Conclusion:(1)Intestinal microecology of the patients with hepatic cirrhosis has changed ,performed that the quantity and proportion of bacterial in mircobial colonies and mircobial membrane has changed. (2)The total quantity in intestinal bacterial of the patients with hepatic cirrhosis are not change significantly.The proportion of bacterial has changed, which lead pH value to increased.(3) There is no significant correlation between the Intestinal microecology and conditional factors(pathogenesis and Child-Pugh classification) among these patients.