AIM: To explore the change of intestinal mucosa barrier function in the progress of non-alcoholic steatohepatitis (NASH) in rats. METHODS: Thirty-two Sprague-Dawley (SD) rats were randomly divided into control group a...AIM: To explore the change of intestinal mucosa barrier function in the progress of non-alcoholic steatohepatitis (NASH) in rats. METHODS: Thirty-two Sprague-Dawley (SD) rats were randomly divided into control group and model group. Rats in the control group were given normal diet, and rats in the model group were given fat-rich diet. Eight rats in each group were killed at end of the 8th and 12th wk, respectively. The levels of endotoxin, D-xylose, TG, TC, ALT and AST, intestinal tissue SOD and MDA as well as intestinal mucus secretory IgA (sIgA) were measured. The pathology of liver was observed by HE staining. RESULTS: At end of the 8th wk, there was no marked difference in the levels of endotoxin, D-xylose and sIgA between the two groups. At end of the 12th wk, rats in the model group developed steatohepatitis and had a higher serum level of endotoxin (P = 0.01) and D-xylose (P = 0.00) and a lower serum level of sIgA (P = 0.007). CONCLUSION: Intestinal mucosa barrier malfunction may exist in NASH rats and may be an important promoter of NASH in rats.展开更多
AIM: To explore the bioactivity of an ethanolic extract of Schizandra arisanensis (SA-Et) and isolated constituents against interleukin-1 β and interferon-γ-mediated β cell death and abolition of insulin secretion....AIM: To explore the bioactivity of an ethanolic extract of Schizandra arisanensis (SA-Et) and isolated constituents against interleukin-1 β and interferon-γ-mediated β cell death and abolition of insulin secretion. METHODS: By employing BRIN-BD11 cells, the effects of SA-Et administration on cytokine-mediated cell death and abolition of insulin secretion were evaluated by a viability assay, cell cycle analysis, and insulin assay. The associated gene and protein expressions were also measured. In addition, the bioactivities of several peak compounds collected from the SA-Et were tested against cytokine-mediated β cell death.RESULTS: Our Results revealed that SA-Et dose-dependently ameliorated cytokine-mediated β cell death and apoptosis. Instead of suppressing inducible nitric oxide synthase/nitric oxide cascade or p38MAPK activity, suppression of stress-activated protein kinase/c-Jun NH2-terminal kinase activity appeared to be the target for SA-Et against the cytokine mix. In addition, SA-Et provided some insulinotropic effects which re-activated the abolished insulin exocytosis in cytokine-treated BRIN-BD11 cells. Finally, schiarisanrin A and B isolated from the SA-Et showed a dose-dependent protective effect against cytokine-mediated β cell death. CONCLUSION: This is the first report on SA-Et ameliorating cytokine-mediated β cell death and dysfunction via anti-apoptotic and insulinotropic actions.展开更多
We developed a Gaussia luciferase (Gluc) reporter replicon of West Nile virus (WNV) and used it to quantify viral translation and RNA replication. The advantage of the Gluc replicon is that Gaussia luciferase is secre...We developed a Gaussia luciferase (Gluc) reporter replicon of West Nile virus (WNV) and used it to quantify viral translation and RNA replication. The advantage of the Gluc replicon is that Gaussia luciferase is secreted into the culture medium from cells transfected with Gluc replicon RNA, and the medium can be assayed directly for luciferase activity. Using a known Flavivirus inhibitor (NITD008), we demonstrated that the Gluc-WNV replicon could be used for antiviral screening. The Gluc-WNV-Rep will be useful for research in antiviral drug development programs, as well as for studying viral replication and pathogenesis of WNV.展开更多
Objective To construct the recombinant adenovirus vector carrying human growth hormone secretagogue receptor type 1a （GHS-R1a） ,for genetic transfection.Methods The full-length human GHS-R1a gene was obtained by PCR...Objective To construct the recombinant adenovirus vector carrying human growth hormone secretagogue receptor type 1a （GHS-R1a） ,for genetic transfection.Methods The full-length human GHS-R1a gene was obtained by PCR amplification and then cloned into the shuttle plasmid pAdTrack-CMV.The linearized plasmid pAdTrack-CMV-GHS-R1a was co-transformed into Escherichia coli （E.coli） BJ5183 cells along with an adenoviral backbone plasmid pAdEasy1.The HEK293 cells were then infected with adenoviruses.The expression of GHS-R1a was indicated by green fluorescent protein （GFP） ,and confirmed by Reverse Transcription Polymerase Chain Reaction （RT-PCR） and Western blot.Results Enzymatic digestion of pAdGHS-R1a yielded a large fragment （approximately 30 kb） and a small fragment （4.5 kb） ,indicating the success-ful construction of recombinant adenovirus expression vector.Expression of GFP was observed by confocal laser scanning microscopy at 24 h after infection.RT-PCR and Western blot further confirmed that GHS-R1a was efficiently expressed in 293 cells.Conclusion Recombinant adenovirus （AdGHS-R1a） is successfully constructed,and the target gene can be expressed efficiently in 293 cells,which provide a valuable tool for further studying the function of GHS-R1a.展开更多
文摘AIM: To explore the change of intestinal mucosa barrier function in the progress of non-alcoholic steatohepatitis (NASH) in rats. METHODS: Thirty-two Sprague-Dawley (SD) rats were randomly divided into control group and model group. Rats in the control group were given normal diet, and rats in the model group were given fat-rich diet. Eight rats in each group were killed at end of the 8th and 12th wk, respectively. The levels of endotoxin, D-xylose, TG, TC, ALT and AST, intestinal tissue SOD and MDA as well as intestinal mucus secretory IgA (sIgA) were measured. The pathology of liver was observed by HE staining. RESULTS: At end of the 8th wk, there was no marked difference in the levels of endotoxin, D-xylose and sIgA between the two groups. At end of the 12th wk, rats in the model group developed steatohepatitis and had a higher serum level of endotoxin (P = 0.01) and D-xylose (P = 0.00) and a lower serum level of sIgA (P = 0.007). CONCLUSION: Intestinal mucosa barrier malfunction may exist in NASH rats and may be an important promoter of NASH in rats.
基金Supported by National Science Council, No. NSC94-2314-B-077-001, No. NSC101-2320-B-077-003-MY2National Research Institute of Chinese Medicine, No. NRICM95-DHM-04
文摘AIM: To explore the bioactivity of an ethanolic extract of Schizandra arisanensis (SA-Et) and isolated constituents against interleukin-1 β and interferon-γ-mediated β cell death and abolition of insulin secretion. METHODS: By employing BRIN-BD11 cells, the effects of SA-Et administration on cytokine-mediated cell death and abolition of insulin secretion were evaluated by a viability assay, cell cycle analysis, and insulin assay. The associated gene and protein expressions were also measured. In addition, the bioactivities of several peak compounds collected from the SA-Et were tested against cytokine-mediated β cell death.RESULTS: Our Results revealed that SA-Et dose-dependently ameliorated cytokine-mediated β cell death and apoptosis. Instead of suppressing inducible nitric oxide synthase/nitric oxide cascade or p38MAPK activity, suppression of stress-activated protein kinase/c-Jun NH2-terminal kinase activity appeared to be the target for SA-Et against the cytokine mix. In addition, SA-Et provided some insulinotropic effects which re-activated the abolished insulin exocytosis in cytokine-treated BRIN-BD11 cells. Finally, schiarisanrin A and B isolated from the SA-Et showed a dose-dependent protective effect against cytokine-mediated β cell death. CONCLUSION: This is the first report on SA-Et ameliorating cytokine-mediated β cell death and dysfunction via anti-apoptotic and insulinotropic actions.
基金supported by the National Natural Science Foundation of China (grant No. 31170158 and 31000090)the '100 Talents Project' of Chinese Academy of Sciences, China (grant No. Y002171YC1)
文摘We developed a Gaussia luciferase (Gluc) reporter replicon of West Nile virus (WNV) and used it to quantify viral translation and RNA replication. The advantage of the Gluc replicon is that Gaussia luciferase is secreted into the culture medium from cells transfected with Gluc replicon RNA, and the medium can be assayed directly for luciferase activity. Using a known Flavivirus inhibitor (NITD008), we demonstrated that the Gluc-WNV replicon could be used for antiviral screening. The Gluc-WNV-Rep will be useful for research in antiviral drug development programs, as well as for studying viral replication and pathogenesis of WNV.
基金supported by the grants from the National Basic Research Development Program of China(No.2007CB516701,2006CB500704)the National Natural Science Foundation of China(No.30770757)
文摘Objective To construct the recombinant adenovirus vector carrying human growth hormone secretagogue receptor type 1a （GHS-R1a） ,for genetic transfection.Methods The full-length human GHS-R1a gene was obtained by PCR amplification and then cloned into the shuttle plasmid pAdTrack-CMV.The linearized plasmid pAdTrack-CMV-GHS-R1a was co-transformed into Escherichia coli （E.coli） BJ5183 cells along with an adenoviral backbone plasmid pAdEasy1.The HEK293 cells were then infected with adenoviruses.The expression of GHS-R1a was indicated by green fluorescent protein （GFP） ,and confirmed by Reverse Transcription Polymerase Chain Reaction （RT-PCR） and Western blot.Results Enzymatic digestion of pAdGHS-R1a yielded a large fragment （approximately 30 kb） and a small fragment （4.5 kb） ,indicating the success-ful construction of recombinant adenovirus expression vector.Expression of GFP was observed by confocal laser scanning microscopy at 24 h after infection.RT-PCR and Western blot further confirmed that GHS-R1a was efficiently expressed in 293 cells.Conclusion Recombinant adenovirus （AdGHS-R1a） is successfully constructed,and the target gene can be expressed efficiently in 293 cells,which provide a valuable tool for further studying the function of GHS-R1a.
