1.Role of Aquaporin 1 on capillary endothelial barrier dysfunction of intestine in rat with experimental acute necrotizing pancreatitis
Yafeng CHEN ; Dianxu FENG ; Jiyan TIAN ; Teng CHEN ; Peihao YIN ; Jinkan XIE ; Wen FENG ; Junkang GUI
Chinese Journal of General Surgery 2012;27(7):576-581
Objective To study the effect of aquaporin 1 on intestinal capillary endothelial barrier in rats with experimental acute necrotizing pancreatitis (ANP).Methods In this study,160 male Sprague-Dawley rats were randomly divided into five groups:Control group ( n =32),ANP group (n =32),NS group,Dexamethasone group,and Acetazolamide group.Eight rats in each group were sacrificed at 3,6,12 and 18 h after induction of experimental models.Volume of ascites and levels of serum amylases were deternined at each time point.Pathological changes in intestine tissues were observed under electron microscope after HE staining.Capillary permeeabilities in intestine tissues were detected by Evans blue (EB) extravasation experiment.The mRNA and protein expressions of AQP1 in intestine tissue were determined by real-time PCR and Western blotting,respectively.Results Serum amylase level in ANP group was significantly higher than that in control group.Amylase level in dexamethasone group was lower than that in ANP group,and amylase level in acetazolamide group was higher than that in ANP group at 12 h (P <0.05 ) ; The concentration of EB in intestine tissues at each time point in ANP group was significantly higher than those in control group,and EB in dexamethasone group was lower than those in ANP group at 6,12 and 18 h.EB in acetazolamide group was higher than that in ANP group at 3 h ( P < 0.05 ) ; The mRNA expression of AQPI in ANP group was significantly lower than that in control group.The expression of AQP1 in dexamethasone group was higher than those in ANP group at 6,12 and 18 h,and the expression of AQP1 in acetazolamide group was lower than that in ANP group at 3,6,12 h in intestine tissue ( P < 0.05 ).Protein expression of AQPI in tissues in ANP group was significantly lower than that in control group.The expression of AQP1 in dexamethasone group increased more than that in ANP group at 3,6,12 h,and the expression of AQP1 in acetazolamide group was lower than that in ANP group at 3 h,6 h ( P < 0.05 ).Conclusions The expression of AQP1 is down-regulated in intestine tissue in rats with acute necrotizing pancreatitis,and AQP1 could play an important role in the pathogenesis of capillary endothelial barrier dysfunction.
2.EXPRESSION OF FOXC2 DURING LYMPHANGIOGENESIS IN HUMAN EMBRYO
Lina TIAN ; Weiguo DENG ; Jiyan ZOU ; Li MA ; Yanliang DING ; Wenling SONG ; Ya FU
Acta Anatomica Sinica 2002;0(06):-
Objective To study the human lymphangiogenesis in the early stage of embryo and the expression of forkhead box C2(FOXC2) in human lymphangiogenesis.Methods Lymphatic vessel endothelial haluronic acid receptor-1(LYVE-1) was used as the special marker of lymphatic vessels.Immunohistochemical and double immunofluorescence staining were used to detect the expressions of FOXC2 and LYVE-1 in lymphatic vessels of 85 human embryos of 5 to 11 weeks pregnancy and analyze the features of lymphatic vessel genesis and development.Results LYVE-1 was initially detected in lymphatic vessels in 7-week human embryos.The lymphatic vessel endothelial cells presented brown staining for LYVE-1 in jugular and thoracic region.LYVE-1 was also found to express in human embryonic mesentery lymphatic vessels on the 70th day of pregnancy.The expression of FOXC2 in human embryo was detected prior to that of LYVE-1.At the beginning of the 6th week of pregnancy,FOXC2 was obviously seen in human embryonic mesoderm mesenchyme.FOXC2 expressed not only in human embryonic lymphatic vessels,but also in the vertebral body,cardiovascular wall and other tissues.The expressions of FOXC2 and LYVE-1 were still seen in human embryonic lymphatic vessel endothelial cells after 80 days of pregnancy.Conclusion Lymphangiogenesis of human embryo begins between the 7th and 8th weeks of pregnancy.FOXC2 and LYVE-1 could have some relation with the human embryonic lymphatic vessel genesis and development.
3.Expression, purification and refolding of extracellular domain of mouse epidermal growth factor receptor.
Bing HU ; Ling TIAN ; You LU ; Li YANG ; Xia ZHAO ; Jiyan LIU ; Yuquan WEI
Journal of Biomedical Engineering 2003;20(2):264-267
A DNA fragment encoding extracellular domain of mouse epidermal growth factor receptor (EGFR) was obtained by PCR from a previous recombinant plasmid. The DNA fragment was then ligated into prokaryotic expression vector, and expressed in Escherichia Coli. The recombinant protein was purified under denature conditions by affinity chromatography, and refolded with gradient dialysis. The recombinant protein could produce antibodies to recognize extracellular domain and full-length of mouse EGFR, and form homodimer in the presence of EGF detected by western blot analysis. These findings provide evidence that the renatured recombinant extracellular domain of mouse epidermal growth factor receptor is immunogenetic and may be important for further application of this protein in functional and immunological research.
Animals
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Escherichia coli
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genetics
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metabolism
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Genetic Vectors
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Mice
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Plasmids
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Receptor, Epidermal Growth Factor
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biosynthesis
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chemistry
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genetics
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isolation & purification
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Transfection