AIM：To examine the expression of intercellular adhesion molecule-1 （ICAM-1） and lymphocyte function-associated antigen-1（LFA-1）expression on canals of Hering （COH）and bile ductules associated with the autoimmun...AIM：To examine the expression of intercellular adhesion molecule-1 （ICAM-1） and lymphocyte function-associated antigen-1（LFA-1）expression on canals of Hering （COH）and bile ductules associated with the autoimmune process of bile duct destruction in primary biliary cirrhosis（PBC）．METHODS：Ten wedged liver biopsies of PBC（five cases each of stages 2 and 3）were studied. The liver specimens were processed for transmission electron microscopy. Immunohistochemistry was performed using anti-ICAM-1 and anti-LFA-1 mouse mAbs.In situ hybridization was done to examine the messenger RNA expression of ICAM-1 in formalin-fixed．paraffin-embedded sections using peptide nucleic acid probes and the catalyzed signal amplification （CSA）technique．Immunogold-silver staining for electron microscopy was Derrormed using anti-ICAM and anti-LFA-1 mouse mAbs．The immunogold particles on epithelial cells of bileductules and cholangiocytes of CoH cells were counted and analyzed semi-quantitatively．Western blotting was performed to confirm ICAM-1 protein expression．RESULTS：In liver tissues of PBC patients．immunohi-stochemistry showed aberrant ICAM-1 expression on the plasma membrane of epithelial cells lining bile ductules，and also on mature cholangiocytes but not on hepatocytes in CoH．LFA-1-positive lymphocytes were closely associated with epithelial cells in bile ductules．ICAM-1 expression at protein level was confirmed by Western blot．In situ hybridization demonstrated ICAM-1 mRNA expression in bile ductules and LFA-1 mRNA in lymphocytes infiltrating the bileductules．By immunoelectron microscopy，ICAM-1 was demonstrated on the basal suface of epithelial cells in bile ductules and on the luminal surfaces of cholangiocytes in damaged COH．Cells with intermediate morphology resembling progenitor cells in Coil were not labeled with ICAM-1 and LFA-1．CONCLUSION：De novo expression of ICAM-1 both on mature cholangiocytes in COH and epithelial cells in bile ductules in PBC implies that lymphocyte-induced destruction through adhesion by ICAM-1 and binding of LFA-1-expressing activated lymphocytes takes place not only in bile ductules but also in the COH．展开更多
Pea seedlings(Pisum sativum L.)were used as materials to test the timings and compartments of hydrogen peroxide(H2O2)triggered by wounding and exogenous jasmonic acid(JA).The results showed that H2O2 could be systemic...Pea seedlings(Pisum sativum L.)were used as materials to test the timings and compartments of hydrogen peroxide(H2O2)triggered by wounding and exogenous jasmonic acid(JA).The results showed that H2O2 could be systemically induced by wounding and exogenous JA.H2O2 increased within 1 h and reached the peak 3―5 h after wounding in either the wounded leaves or the unwounded leaves adjacent to the wounded ones and the inferior leaves far from the wounded ones.After this,H2O2 decreased and recovered to the control level 12 h after wounding.The activities of antioxidant enzymes,however,were rapidly increased by wounding.Diphenylene iodonium(DPI),an inhibitor of NADPH oxidase,could significantly inhibit H2O2 burst that was mediated by wounding and exogenous JA.Assay of H2O2 subcellular location showed that H2O2 in response to wounding and exogenous JA was predominantly accumulated in plasma membrane,cell wall and apoplasmic space.Numerous JA(gold particles)was found via immu-nogold electron microscopy to be located in cell wall and phloem zones of mesophyll cell after wounding.展开更多
文摘AIM：To examine the expression of intercellular adhesion molecule-1 （ICAM-1） and lymphocyte function-associated antigen-1（LFA-1）expression on canals of Hering （COH）and bile ductules associated with the autoimmune process of bile duct destruction in primary biliary cirrhosis（PBC）．METHODS：Ten wedged liver biopsies of PBC（five cases each of stages 2 and 3）were studied. The liver specimens were processed for transmission electron microscopy. Immunohistochemistry was performed using anti-ICAM-1 and anti-LFA-1 mouse mAbs.In situ hybridization was done to examine the messenger RNA expression of ICAM-1 in formalin-fixed．paraffin-embedded sections using peptide nucleic acid probes and the catalyzed signal amplification （CSA）technique．Immunogold-silver staining for electron microscopy was Derrormed using anti-ICAM and anti-LFA-1 mouse mAbs．The immunogold particles on epithelial cells of bileductules and cholangiocytes of CoH cells were counted and analyzed semi-quantitatively．Western blotting was performed to confirm ICAM-1 protein expression．RESULTS：In liver tissues of PBC patients．immunohi-stochemistry showed aberrant ICAM-1 expression on the plasma membrane of epithelial cells lining bile ductules，and also on mature cholangiocytes but not on hepatocytes in CoH．LFA-1-positive lymphocytes were closely associated with epithelial cells in bile ductules．ICAM-1 expression at protein level was confirmed by Western blot．In situ hybridization demonstrated ICAM-1 mRNA expression in bile ductules and LFA-1 mRNA in lymphocytes infiltrating the bileductules．By immunoelectron microscopy，ICAM-1 was demonstrated on the basal suface of epithelial cells in bile ductules and on the luminal surfaces of cholangiocytes in damaged COH．Cells with intermediate morphology resembling progenitor cells in Coil were not labeled with ICAM-1 and LFA-1．CONCLUSION：De novo expression of ICAM-1 both on mature cholangiocytes in COH and epithelial cells in bile ductules in PBC implies that lymphocyte-induced destruction through adhesion by ICAM-1 and binding of LFA-1-expressing activated lymphocytes takes place not only in bile ductules but also in the COH．
基金This work was supported by the National Natural Science Foundation of China(Grant No.30270918)the National High Technology Research and Development Program of China(863 ProgramGrant No.2003AA241170).
文摘Pea seedlings(Pisum sativum L.)were used as materials to test the timings and compartments of hydrogen peroxide(H2O2)triggered by wounding and exogenous jasmonic acid(JA).The results showed that H2O2 could be systemically induced by wounding and exogenous JA.H2O2 increased within 1 h and reached the peak 3―5 h after wounding in either the wounded leaves or the unwounded leaves adjacent to the wounded ones and the inferior leaves far from the wounded ones.After this,H2O2 decreased and recovered to the control level 12 h after wounding.The activities of antioxidant enzymes,however,were rapidly increased by wounding.Diphenylene iodonium(DPI),an inhibitor of NADPH oxidase,could significantly inhibit H2O2 burst that was mediated by wounding and exogenous JA.Assay of H2O2 subcellular location showed that H2O2 in response to wounding and exogenous JA was predominantly accumulated in plasma membrane,cell wall and apoplasmic space.Numerous JA(gold particles)was found via immu-nogold electron microscopy to be located in cell wall and phloem zones of mesophyll cell after wounding.
