Localization and estimation of histamine(HA)content in the wound edge in 81cases of SD rats were carried out by microfluorometric method specific for this aminewhich forms a complex with ophthalaedehyde(OPT).At the same time,thedistribution and the density of the mast calls in the same areas were observed byToluidine blue stain.in all skin specimens with antemortem wounds,both the epidermisand upper dermis exhibited extracellular yellowish fluorescente band of the HA-OPTcomplex.The zone spreaded in wound edge with the lapse of time in antemorteminjuries.The content of HA increased gradually up to 30′,and then yellow hista- mine fluorescence in areas 0-200? extended from wound edge decreased.None of thesefeatures could be observed in normal control skin and postmortemin jured skin.themast cell degranulation could be demonstrated in all antemortem injured skin.Nostatistic relationship between the degranulation of mast cell and the HA-OPTfluorescence existed in either ante-or post-mortem injured groups.This study indi-cated that the skin HA microfluoremetry by OPT method has practical value fordistingushing the ante-from post-mortem wounds and for timing antemortem wounds.

Experimental studies on the myocardial ischemia and reperfusion injury in 16 anaethetized SDrats,of which,8 animals were pretreated with morphine(5 mg/kg,i.p.)for preventing of arrhyth-mias,were studied immunocytochemically with anti-muscle actin specific monoclonal antibody (HHF_(35)),8 shan-operated rats were used as control.With HHF_(35) ABC immunocytochemical method,the area of myocardial ischemia and reperfusion injury(without morphine)showed decrease or ab-sence of staining,large areas of staining loss were also seen.In the group with morphine,only smallfoci of staining absence were shown.The myocardium in control animals showed evenly positive stain-ing.No change were seen with HE staining in all groups.The results obtained with HHF_(35) stainingsupport its important value in studying on myocardic reperfusion injury,and indicated that the degreeof myocardic damage may be relative to the arrhythmias in myocardial reperfusion injury.

OBJECTIVETo investigate the protective effects of exendin-4 on vascular endothelial cells and explore the possible mechanism.

METHODSHuman umbilical vascular endothelial cells (HUVECs) were cultured in the presence of high glucose and tumor necrosis factor-α (TNF-α, 10 ng/ml) with or without exendin-4. The level of nitric oxide (NO) in the cell culture supernatant was measured using a nitrate reductase method. The expression of intercellular adhesion molecule-1 (ICAM-1) mRNA was measured by real-time PCR, and nuclear factor-κB (NF-κB) p65 translocation was detected using immunofluorescence assay. Western blotting was employed to measure the expression of p38 MAPK protein in the treated cells.

RESULTSIn the presence of high glucose and TNF-α, treatment of cells with exendin-4 did not obviously affect the cellular synthesis of NO, but significantly down-regulated the expression of ICAM-1 mRNA (P<0.01). The nuclear fluorescence intensity of NF-κB p65 and the expression level of p38 MAPK protein in the cells were significantly lowered by exendin-4 treatment (P<0.01).

CONCLUSIONExendin-4 ameliorates high glucose- and TNF-α-induced HUVEC-12 cell damage by inhibiting the expression of p38 MAPK protein and translocation of NF-κB p65.

Cell Line ; Culture Media ; chemistry ; Glucose ; adverse effects ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Peptides ; pharmacology ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; adverse effects ; Venoms ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism

Pathogenesis of the abdominal aortic aneurysm has been attributed to neovascularization of the aortic wall. However, it is not clear whether angiogenesis persists in the aneurysm. In sections of aneurysms, we determined the immunohistochemical distributions of the alpha v beta 3 integrin, tenascin and endothelial nitric oxide synthase (eNOS), which are markers respectively, of angiogenesis, matrix remodeling and vasoregulatory function. In addition, we used reverse transcription followed by in situ PCR, to determine the distribution of alpha v mRNA. All aneurysm specimens exhibited extensive increases of wall vascularization as compared with the control aortic wall, and showed the presence of perivascular inflammatory exudates containing macrophages and lymphocytes. The neovascularization consisted of thick-walled vessels in the media and adventitia, and capillaries in the subintima. The majority of vessels stained positively for the alpha v beta 3 antigen and eNOS. Tenascin was deposited as bands that circumscribed thick-walled vessels. The distribution of av mRNA was extensive and was positive even in those vessels that failed to stain for the alpha v beta 3 protein. No staining was evident in control aortas for the alpha v beta 3 antigen, tenascin or alpha v mRNA. The upregulation of av mRNA and the alpha v beta 3 integrin in blood vessels surrounded by a matrix expressing tenascin, indicates that angiogenesis is an ongoing process in the mature aortic aneurysm.
Adult ; Aorta, Abdominal/immunology/pathology ; Aortic Aneurysm, Abdominal/*pathology ; Biological Markers/analysis/metabolism ; Female ; Humans ; Integrin alphaVbeta3/analysis/genetics/metabolism ; Male ; Neovascularization, Pathologic/genetics/*metabolism ; Nitric-Oxide Synthase/analysis/metabolism ; RNA, Messenger/analysis/metabolism ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, P.H.S. ; Tenascin/analysis/metabolism