Actual sandstone micromodel was used in this work to conduct the microscopic waterflooding experiment of ultra-low sandstone reservoir,since the inside seepage characteristics of microscopic waterflooding process of C...Actual sandstone micromodel was used in this work to conduct the microscopic waterflooding experiment of ultra-low sandstone reservoir,since the inside seepage characteristics of microscopic waterflooding process of Chang 8 ultra-low permeability sandstone reservoir of Upper Triassic Yanchang formation in Huaqing region of the Ordos Basin,China is difficult to observe directly.Combined with physical property,casting thin sections,constant-rate mercury injection capillary pressure and nuclear magnetic resonance,the influence of reservoir property on the waterflooding characteristics in pores were analyzed and evaluated.Seepage paths of waterflooding characteristics were divided into four types:homogeneous seepage,reticular-homogeneous seepage,finger-reticular seepage and finger-like seepage,the waterflooding efficiency of which decreases in turn.More than 70%of residual oil occurs as flowing-around seepage and oil film.Physical property,pore structure and movable fluid characteristics are all controlled by digenesis and their impacts on waterflooding efficiency are in accordance.Generally,the pore throat radius size and distribution and movable fluid percentage are closely related to waterflooding law.展开更多
To determine the role of corticotropin releasing factor receptor (CRF2) in epithelial permeability and enterocyte cell differentiation.METHODSFor this purpose, we used rat Sprague Dawley and various colon carcinoma ce...To determine the role of corticotropin releasing factor receptor (CRF2) in epithelial permeability and enterocyte cell differentiation.METHODSFor this purpose, we used rat Sprague Dawley and various colon carcinoma cell lines (SW620, HCT8R, HT-29 and Caco-2 cell lines). Expression of CRF2 protein was analyzed by fluorescent immunolabeling in normal rat colon and then by western blot in dissociated colonic epithelial cells and in the lysates of colon carcinoma cell lines or during the early differentiation of HT-29 cells (ten first days). To assess the impact of CRF2 signaling on colonic cell differentiation, HT-29 and Caco-2 cells were exposed to Urocortin 3 recombinant proteins (Ucn3, 100 nmol/L). In some experiments, cells were pre-exposed to the astressin 2b (A2b) a CRF2 antagonist in order to inhibit the action of Ucn3. Intestinal cell differentiation was first analyzed by functional assays: the trans-cellular permeability and the para-cellular permeability were determined by Dextran-FITC intake and measure of the transepithelial electrical resistance respectively. Morphological modifications associated to epithelial dysfunction were analyzed by confocal microscopy after fluorescent labeling of actin (phaloidin-TRITC) and intercellular adhesion proteins such as E-cadherin, p120ctn, occludin and ZO-1. The establishment of mature adherens junctions (AJ) was monitored by following the distribution of AJ proteins in lipid raft fractions, after separation of cell lysates on sucrose gradients. Finally, the mRNA and the protein expression levels of characteristic markers of intestinal epithelial cell (IEC) differentiation such as the transcriptional factor krüppel-like factor 4 (KLF4) or the dipeptidyl peptidase IV (DPPIV) were performed by RT-PCR and western blot respectively. The specific activities of DPPIV and alkaline phosphatase (AP) enzymes were determined by a colorimetric method.RESULTSCRF2 protein is preferentially expressed in undifferentiated epithelial cells from the crypts of colon and in human colon carcinoma cell lines. Furthermore, CRF2 expression is down regulated according to the kinetic of HT-29 cell differentiation. By performing functional assays, we found that Ucn3-induced CRF2 signaling alters both para- and trans-cellular permeability of differentiated HT-29 and Caco-2 cells. These effects are partly mediated by Ucn3-induced morphological changes associated with the disruption of mature AJ in HT-29 cells and tight junctions (TJ) in Caco-2 cells. Ucn3-mediated activation of CRF2 decreases mRNA and protein expression levels of KLF4 a transcription factor involved in IEC differentiation. This signaling is correlated to a down-regulation of key IEC markers such as DPPIV and AP, at both transcriptional and post-transcriptional levels.CONCLUSIONOur findings suggest that CRF2 signaling could modulate IEC differentiation. These mechanisms could be relevant to the stress induced epithelial alterations found in inflammatory bowel diseases.展开更多
基金Project(2015KTCL01-09)supported by the Innovation Project of Science and Technology of Shaanxi Province,ChinaProject(2015M582699)supported by the China Postdoctoral Science Foundation+1 种基金Project(2016JQ4022)supported by the Natural Science Foundation Research Project of Shaanxi Province,ChinaProject(41702146)supported by the National Natural Science Foundation of China
文摘Actual sandstone micromodel was used in this work to conduct the microscopic waterflooding experiment of ultra-low sandstone reservoir,since the inside seepage characteristics of microscopic waterflooding process of Chang 8 ultra-low permeability sandstone reservoir of Upper Triassic Yanchang formation in Huaqing region of the Ordos Basin,China is difficult to observe directly.Combined with physical property,casting thin sections,constant-rate mercury injection capillary pressure and nuclear magnetic resonance,the influence of reservoir property on the waterflooding characteristics in pores were analyzed and evaluated.Seepage paths of waterflooding characteristics were divided into four types:homogeneous seepage,reticular-homogeneous seepage,finger-reticular seepage and finger-like seepage,the waterflooding efficiency of which decreases in turn.More than 70%of residual oil occurs as flowing-around seepage and oil film.Physical property,pore structure and movable fluid characteristics are all controlled by digenesis and their impacts on waterflooding efficiency are in accordance.Generally,the pore throat radius size and distribution and movable fluid percentage are closely related to waterflooding law.
基金Supported by grants from Association pour la Recherche sur le Cancer,Ligue Nationale contre le Cancer,No.GEFLUC and No.ESPOIR
文摘To determine the role of corticotropin releasing factor receptor (CRF2) in epithelial permeability and enterocyte cell differentiation.METHODSFor this purpose, we used rat Sprague Dawley and various colon carcinoma cell lines (SW620, HCT8R, HT-29 and Caco-2 cell lines). Expression of CRF2 protein was analyzed by fluorescent immunolabeling in normal rat colon and then by western blot in dissociated colonic epithelial cells and in the lysates of colon carcinoma cell lines or during the early differentiation of HT-29 cells (ten first days). To assess the impact of CRF2 signaling on colonic cell differentiation, HT-29 and Caco-2 cells were exposed to Urocortin 3 recombinant proteins (Ucn3, 100 nmol/L). In some experiments, cells were pre-exposed to the astressin 2b (A2b) a CRF2 antagonist in order to inhibit the action of Ucn3. Intestinal cell differentiation was first analyzed by functional assays: the trans-cellular permeability and the para-cellular permeability were determined by Dextran-FITC intake and measure of the transepithelial electrical resistance respectively. Morphological modifications associated to epithelial dysfunction were analyzed by confocal microscopy after fluorescent labeling of actin (phaloidin-TRITC) and intercellular adhesion proteins such as E-cadherin, p120ctn, occludin and ZO-1. The establishment of mature adherens junctions (AJ) was monitored by following the distribution of AJ proteins in lipid raft fractions, after separation of cell lysates on sucrose gradients. Finally, the mRNA and the protein expression levels of characteristic markers of intestinal epithelial cell (IEC) differentiation such as the transcriptional factor krüppel-like factor 4 (KLF4) or the dipeptidyl peptidase IV (DPPIV) were performed by RT-PCR and western blot respectively. The specific activities of DPPIV and alkaline phosphatase (AP) enzymes were determined by a colorimetric method.RESULTSCRF2 protein is preferentially expressed in undifferentiated epithelial cells from the crypts of colon and in human colon carcinoma cell lines. Furthermore, CRF2 expression is down regulated according to the kinetic of HT-29 cell differentiation. By performing functional assays, we found that Ucn3-induced CRF2 signaling alters both para- and trans-cellular permeability of differentiated HT-29 and Caco-2 cells. These effects are partly mediated by Ucn3-induced morphological changes associated with the disruption of mature AJ in HT-29 cells and tight junctions (TJ) in Caco-2 cells. Ucn3-mediated activation of CRF2 decreases mRNA and protein expression levels of KLF4 a transcription factor involved in IEC differentiation. This signaling is correlated to a down-regulation of key IEC markers such as DPPIV and AP, at both transcriptional and post-transcriptional levels.CONCLUSIONOur findings suggest that CRF2 signaling could modulate IEC differentiation. These mechanisms could be relevant to the stress induced epithelial alterations found in inflammatory bowel diseases.
