AIM： To evaluate the expression of fibrinogenlike protein 2 （fgl2） and its correlation with disease progression in both mice and patients with severe viral hepatitis. METHODS： Balb/cJ or A/J mice were infected int...AIM： To evaluate the expression of fibrinogenlike protein 2 （fgl2） and its correlation with disease progression in both mice and patients with severe viral hepatitis. METHODS： Balb/cJ or A/J mice were infected intraperitoneally （ip） with 100 PFU of murine hepatitis virus type 3 （MHV-3）, liver and serum were harvested at 24, 48, and 72 h post infection for further use. Liver tissues were obtained from 23 patients with severe acute chronic （AOC） hepatitis B and 13 patients with mild chronic hepatitis B. Fourteen patients with mild chronic hepatitis B with cirrhosis and 4 liver donors served as normal controls. In addition, peripheral blood mononuciear cells （PBMC） were isolated from 30 patients （unpaired） with severe AOC hepatitis B and 10 healthy volunteers as controls. Procoagulant activity representing functional prothrombinase activity in PBMC and white blood cells was also assayed. A polyclonal antibody against fgl2 was used to detect the expression of both mouse and human fgl2 protein in liver samples as well as in PBMC by immunohistochemistry staining in a separate set of studies. Alanine aminotransferase （ALT） and total bilirubin （TBil） in serum were measured to assess the severity of liver injury.RESULTS： Histological changes were found in liver sections 12-24 h post MHV-3 infection in Balb/cJ mice. In association with changes in liver histology, marked elevations in serum ALT and TBil were observed. House fgl2 （mfgl2） protein was detected in the endothelium of intrahepatic veins and hepatic sinusoids within the liver 24 h after MHV-3 infection. Liver tissues from the patients with severe AOC hepatitis B had classical pathological features of acute necroinflammation. Human fgl2 （hfgl2） was detected in 21 of 23 patients （91.30%） with severe AOC hepatitis B, while only 1 of 13 patients （7.69%） with mild chronic hepatitis B and cirrhosis had hfgl2 mRNA or protein expression. Twenty-eight of thirty patients （93.33%） with severe AOC hepatitis B and 1 of 展开更多
目的探讨补体C5a/C5aR通路在结肠癌发病中对纤维介素蛋白-2凝血酶原酶(fibrinogen-like protein 2,FGL2)的调节机制,明确补体系统在结肠癌发病中的功能和作用。方法收集2013年12月至2015年7月第三军医大学西南医院普外科、新桥医院消化...目的探讨补体C5a/C5aR通路在结肠癌发病中对纤维介素蛋白-2凝血酶原酶(fibrinogen-like protein 2,FGL2)的调节机制,明确补体系统在结肠癌发病中的功能和作用。方法收集2013年12月至2015年7月第三军医大学西南医院普外科、新桥医院消化科15例经病理活检确诊的住院临床结肠癌患者癌及癌旁组织。采用免疫组织化学方法检测结肠癌患者癌及癌旁组织中补体活化片段C5b-9、活化C3及C5aR的表达,佐证补体系统在结肠癌中的活化状态;进一步通过体内外实验(Western blot)验证结肠癌患者癌及癌旁组织MAPK信号通路中P38的磷酸化水平及FGL2的沉积,明确结肠癌发病中补体C5a/C5aR通路与二者的调节关系。结果免疫组织化学及Western blot实验证实结肠癌患者癌组织C5b-9、活化C3及C5aR的表达明显高于癌旁组织,FGL2的沉积亦显著高于癌旁组织;体外结果提示C5a能促进巨噬细胞系Raw264.7MAPK通路中P38的磷酸化水平(P=0.0013)及FGL2的产生能力(P=0.0071),加入C5aR阻断剂之后,二者表达随之减弱。结论 C5a/C5aR通路能通过MAPK信号途径中的P38信号完成对FGL2的调节作用进而参与结肠癌的发病进程。展开更多
Objective:To study the mechanism and significance of pH change in the coronary artery microthrombosis of rats.Methods:After the sodium laurate-induced model of coronary artery microthrombosis of rats was constructed,t...Objective:To study the mechanism and significance of pH change in the coronary artery microthrombosis of rats.Methods:After the sodium laurate-induced model of coronary artery microthrombosis of rats was constructed,the vascular endothelial cells were separated and then cultured in the mediums with different pH values for 24 h.Enzyme linked immunosorbent assay was used to detect the content of von Willebrand factor(vWF) in the medium;while the real-time PCR and western blot assay were used to detect the expression of fibrinogenlike protein 2(FGL2) at the mRNA and protein level.The comprehensive evaluation was performed to discuss the effect of pH change on the coronary artery microthrombosis of rats.Results:The expression level of vWF detected by enzyme linked immunosorbent assay was 336.67±24.95,311.33±14.98,359.67±39.63,354.67±49.01 and 332.00±33.42(pg/mL) respectively;while the expression of vWF in the model group was 570.00±57.94,524.67±57.94,437.00±95.38,415.33±44.38 and 444.67±74.31 respectively.Being cultured under the different pH values,the relative expression level of FGL2 mRNA in the model group was 7.93±0.93,6.70±0.70.5.03±0.32,5.13±0:40 and 5.57±0.83 respectively.Conclusions:The coronary artery microthrombosis of rats can cause the high expression and secretion of vWF.Meanwhile,FGL2 is also up-regulated in the thrombosis and such up-regulation is more significant in the condition with low pH,which indicates that the low-pH condition may be one of factors that contribute to the cardiovascular diseases.展开更多
CD4 + CD25 + CD127 dim/regulatory T cells(Tregs) have been implicated in suppressing T cell immune responses to hepatitis B virus(HBV),but the inhibition mechanism has not being clear yet.This study investigated the e...CD4 + CD25 + CD127 dim/regulatory T cells(Tregs) have been implicated in suppressing T cell immune responses to hepatitis B virus(HBV),but the inhibition mechanism has not being clear yet.This study investigated the effects of soluble FGL2(sFGL2) secreted by Tregs on immune suppression in chronic HBV-infected patients.We verified that sFGL2 protein and mRNA were highly expressed in Tregs.The separated Tregs by using magnetic beads from peripheral blood mononuclear cells(PBMCs) in 20 patients with chronic hepatitis B were co-cultured with PBMCs at a ratio of 1:3 with anti-CD3 stimulating antibody or FGL2 blocking antibody.The proliferation index of CD8 + T cells after blocking FGL2 was higher than that in blank group(3.58±0.18 vs.3.28±0.17,P=0.034) in 18 of 20 samples,and lower than that in CD3 stimulation group(3.82±0.19,P=0.026) in 16 of 20 samples.The IFN-γ secreted in the mixed culture in the absence of Tregs was higher than that in the culture in the presence of Tregs,but it could be abolished by FGL2 blocking antibody.These results suggest that sFGL2 protein secreted by Tregs suppresses the proliferation and function of CD8 + T cells in chronic hepatitis B.展开更多
基金Supported by the National Natural Science Foundation of China for Distinguished Young Scholars, No. 30225040 for Dr Ning Q,No. 30123019 for Dr Luo XP
文摘AIM： To evaluate the expression of fibrinogenlike protein 2 （fgl2） and its correlation with disease progression in both mice and patients with severe viral hepatitis. METHODS： Balb/cJ or A/J mice were infected intraperitoneally （ip） with 100 PFU of murine hepatitis virus type 3 （MHV-3）, liver and serum were harvested at 24, 48, and 72 h post infection for further use. Liver tissues were obtained from 23 patients with severe acute chronic （AOC） hepatitis B and 13 patients with mild chronic hepatitis B. Fourteen patients with mild chronic hepatitis B with cirrhosis and 4 liver donors served as normal controls. In addition, peripheral blood mononuciear cells （PBMC） were isolated from 30 patients （unpaired） with severe AOC hepatitis B and 10 healthy volunteers as controls. Procoagulant activity representing functional prothrombinase activity in PBMC and white blood cells was also assayed. A polyclonal antibody against fgl2 was used to detect the expression of both mouse and human fgl2 protein in liver samples as well as in PBMC by immunohistochemistry staining in a separate set of studies. Alanine aminotransferase （ALT） and total bilirubin （TBil） in serum were measured to assess the severity of liver injury.RESULTS： Histological changes were found in liver sections 12-24 h post MHV-3 infection in Balb/cJ mice. In association with changes in liver histology, marked elevations in serum ALT and TBil were observed. House fgl2 （mfgl2） protein was detected in the endothelium of intrahepatic veins and hepatic sinusoids within the liver 24 h after MHV-3 infection. Liver tissues from the patients with severe AOC hepatitis B had classical pathological features of acute necroinflammation. Human fgl2 （hfgl2） was detected in 21 of 23 patients （91.30%） with severe AOC hepatitis B, while only 1 of 13 patients （7.69%） with mild chronic hepatitis B and cirrhosis had hfgl2 mRNA or protein expression. Twenty-eight of thirty patients （93.33%） with severe AOC hepatitis B and 1 of
基金supported by Medical Scientific Research Foundation of Guangdong Province(No.A2012242)
文摘Objective:To study the mechanism and significance of pH change in the coronary artery microthrombosis of rats.Methods:After the sodium laurate-induced model of coronary artery microthrombosis of rats was constructed,the vascular endothelial cells were separated and then cultured in the mediums with different pH values for 24 h.Enzyme linked immunosorbent assay was used to detect the content of von Willebrand factor(vWF) in the medium;while the real-time PCR and western blot assay were used to detect the expression of fibrinogenlike protein 2(FGL2) at the mRNA and protein level.The comprehensive evaluation was performed to discuss the effect of pH change on the coronary artery microthrombosis of rats.Results:The expression level of vWF detected by enzyme linked immunosorbent assay was 336.67±24.95,311.33±14.98,359.67±39.63,354.67±49.01 and 332.00±33.42(pg/mL) respectively;while the expression of vWF in the model group was 570.00±57.94,524.67±57.94,437.00±95.38,415.33±44.38 and 444.67±74.31 respectively.Being cultured under the different pH values,the relative expression level of FGL2 mRNA in the model group was 7.93±0.93,6.70±0.70.5.03±0.32,5.13±0:40 and 5.57±0.83 respectively.Conclusions:The coronary artery microthrombosis of rats can cause the high expression and secretion of vWF.Meanwhile,FGL2 is also up-regulated in the thrombosis and such up-regulation is more significant in the condition with low pH,which indicates that the low-pH condition may be one of factors that contribute to the cardiovascular diseases.
基金supported by National 973 Project in Viral Hepatitis,China (MOST,No. 2007CB512900)
文摘CD4 + CD25 + CD127 dim/regulatory T cells(Tregs) have been implicated in suppressing T cell immune responses to hepatitis B virus(HBV),but the inhibition mechanism has not being clear yet.This study investigated the effects of soluble FGL2(sFGL2) secreted by Tregs on immune suppression in chronic HBV-infected patients.We verified that sFGL2 protein and mRNA were highly expressed in Tregs.The separated Tregs by using magnetic beads from peripheral blood mononuclear cells(PBMCs) in 20 patients with chronic hepatitis B were co-cultured with PBMCs at a ratio of 1:3 with anti-CD3 stimulating antibody or FGL2 blocking antibody.The proliferation index of CD8 + T cells after blocking FGL2 was higher than that in blank group(3.58±0.18 vs.3.28±0.17,P=0.034) in 18 of 20 samples,and lower than that in CD3 stimulation group(3.82±0.19,P=0.026) in 16 of 20 samples.The IFN-γ secreted in the mixed culture in the absence of Tregs was higher than that in the culture in the presence of Tregs,but it could be abolished by FGL2 blocking antibody.These results suggest that sFGL2 protein secreted by Tregs suppresses the proliferation and function of CD8 + T cells in chronic hepatitis B.
