ObjectiveTo explore the variation of the structure of the intestinal flora between healthy people and patients with obstructive jaundice perioperatively. MethodsFrom February 2013 to August 2014, 20 patients with obstructive jaundice and 10 healthy persons (normal control group) in our hospitol were selected as the research object. The first stool specimens of the research object after admission were obtained and the total fecal bacteria DNA were extracted. After polymerase chain reaction amplification, the changes in the structure of bacterial flora were dynamic observed by using denaturing gradient gel electrophoresis (DGGE), and the gel bands were analyzed by using Quantity One software. The similarity and diversity of flora structure, and principal component analysis (PCA) were analyzed. ResultsSignificant differences of colonic microflora were found between patients with obstructive jaundice and healthy people; advantage intestinal flora in obstructive jaundice patients was significant lower than the normal control group. With the extension of time and degree of obstruction aggravated, a descending trend was found in number, abundance, and diversity of the intestinal microflora (P < 0.05). ConclusionThere is significant differences in the structure of colon bacteria in patients with obstructive jaundice and healthy persons.
ObjectiveTo explore the influence of miRNA-155/PU.1 signaling pathway blockade on bone marrow-derived dendritic cells (DCs) maturation and immune function of rat small intestinal transplantation. MethodsThe DCs were induced by adherent culture.The critical transcription factor gene PU.1 was designed and PU.1 siRNA was synthe-sized.The DCs were transfected by liposome transfection and a pair of PU.1 siRNA was screened according to the high silencing efficiency.The expressions of DCs surface markers CD80, CD86, and MHC-Ⅱamong three groups (PU.1 silent group, negative control group, and control group) were analyzed by flow cytometry.The IL-10 and IL-12p70 secretion levels in the supernatant were tested by ELISA method.The allogeneic T lymphocyte proliferation was tested by mixed lymphocyte reaction.The transfected cells were intravenously injected into the recipient rat on day 7 before intestinal transplantation.The survival conditions as well as pathological changes were observed in each group recipients. Results①The surface molecules CD80, CD86, and MHC-Ⅱin the PU.1 silent group were (27.0±5.6)%, (23.6±4.8)%, and (36.8±6.8)%, respectively; versus (74.0±9.4)%, (76.5±8.7)%, and (87.8±11.3)% in the negative control group, respectively, which were significantly lower in former and showing an in creased trend (P < 0.05).②Compared with the negative control group, IL-10 secretion level was significantly increased (P < 0.05), IL-12p70 secretion level significantly decreased (P < 0.05) in the PU.1 silent group.③The proliferation of T lymphocytes in the PU.1 silent group was significantly lower than that in the negative control group (P < 0.05).④When the transfected DCs were injected into the intestinal transplantation rats on day 7 before operation, the survival time was (14.3±3.3) d, (7.8±1.5) d, and (8.0±2.5) d in the PU.1 silent group, negative control group, and control group, respectively, which in the PU.1 silent group were significantly longer than that in the other two groups (P < 0.05), and the graft pathology showed that there were mild intestinal tissue damage, lymphocyte infiltration or villus edema in the PU.1 silent group. ConclusionmiRNA-155/PU.1 signaling pathway blockade could reduce DCs maturation and induce receptor-specific immune tolerance, which are proved both in vivo and in vitro.