AIM: To establish a simple method to quantify lipid classes in liver diseases and to decipher the lipid profile in p62/IMP2-2/IGF2BP2-2 transgenic mice.METHODS: Liver-specific overexpression of the insulin-like growth...AIM: To establish a simple method to quantify lipid classes in liver diseases and to decipher the lipid profile in p62/IMP2-2/IGF2BP2-2 transgenic mice.METHODS: Liver-specific overexpression of the insulin-like growth factor 2 mRNA binding protein p62/IMP2-2/IGF2BP2-2 was used as a model for steatosis.Steatohepatitis was induced by feeding a methioninecholine deficient diet. Steatosis was assessed histologically. For thin layer chromatographic analysis, lipids were extracted from freeze-dried tissues by hexane/2-propanol, dried, redissolved, and chromatographically separated by a two-solvent system. Dilution series of lipid standards were chromatographed, detected, andquantified. The detection was performed by either2',7'-dichlorofluoresceine or a sulfuric acid/ethanol mixture.RESULTS: Histological analyses confirmed steatosis and steatohepatitis development. The extraction,chromatographic, and detection method showed high inter-assay reproducibility and allowed quantification of the different lipid classes. The analyses confirmed an increase of triglycerides and phosphatidylethanolamine and a decrease in phosphatidylcholine in the methionine-choline deficient diet. The method was used for the first time to asses the lipid classes induced in the p62-overexpressing mouse model and showed a significant increase in all detected lipid species with a prominent increase of triglycerides by 2-fold. Interestingly, the ratio of phosphatidylcholine to phosphatidylethanolamine was decreased, as previously suggested as a marker in the progression from steatosis to steatohepatitis.CONCLUSION: The thin layer chromatography analysis allows a reliable quantification of lipid classes and provides detailed insight into the lipogenic effect of p62.展开更多
[ Objective] To investigate the effects of concentrate/roughage ratios and nitrogen levels on in-vitro rumen fermentation of urea-treating corn stalk. [ Method] The concentrate/roughage ratios were 7： 3, 5：5 and 3：...[ Objective] To investigate the effects of concentrate/roughage ratios and nitrogen levels on in-vitro rumen fermentation of urea-treating corn stalk. [ Method] The concentrate/roughage ratios were 7： 3, 5：5 and 3： 7. The crude protein levels were 11% and 14%. The in-vitro culture time was 2, 4, 6 and 48 h. [ Result] The pH value of broth decreased significantly with the decline in the proportion of roughage （ P 〈 0.01 ） and with the increase in the crude protein levels （P 〈 0.05）. The ammonium nitrogen concentration and acetic acid/propionic acid ratio of the fermen- ted products decreased with the increase in the crude protein levels and proportion of concentrate in diet. The diet with concentrate/roughage ratio of 5：5 and crude protein level of 14% had significantly higher digestibility of dry matter and organic matter than other diets （ P 〈 0.05）. The digesti- bility of organic matter increased gradually with the increasing proportion of concentrate and crude protein level. With the increase in the proportion of concentrate, the microbial protein levels increased remarkably, while the acetic acid/propionic acid ratio declined. [ Conclusion] The concentrate/ roughage ratios and nitrogen levels affect rumen fermentation and microbial growth during in-vitro culture. However, the best supplementary feeding results of urea-treatinq corn stalks can be obtained when the concentrate/rouahaae ratio is below 5：5 and the crude protein level is 14%.展开更多
基金Supported by The Graduiertenf rderung of Saarland University(Laggai S)an EASL Dame Sheila Sherlock Fellowship(Kessler SM)the research committee of Saarland University(61-cl/Anschub2012)
文摘AIM: To establish a simple method to quantify lipid classes in liver diseases and to decipher the lipid profile in p62/IMP2-2/IGF2BP2-2 transgenic mice.METHODS: Liver-specific overexpression of the insulin-like growth factor 2 mRNA binding protein p62/IMP2-2/IGF2BP2-2 was used as a model for steatosis.Steatohepatitis was induced by feeding a methioninecholine deficient diet. Steatosis was assessed histologically. For thin layer chromatographic analysis, lipids were extracted from freeze-dried tissues by hexane/2-propanol, dried, redissolved, and chromatographically separated by a two-solvent system. Dilution series of lipid standards were chromatographed, detected, andquantified. The detection was performed by either2',7'-dichlorofluoresceine or a sulfuric acid/ethanol mixture.RESULTS: Histological analyses confirmed steatosis and steatohepatitis development. The extraction,chromatographic, and detection method showed high inter-assay reproducibility and allowed quantification of the different lipid classes. The analyses confirmed an increase of triglycerides and phosphatidylethanolamine and a decrease in phosphatidylcholine in the methionine-choline deficient diet. The method was used for the first time to asses the lipid classes induced in the p62-overexpressing mouse model and showed a significant increase in all detected lipid species with a prominent increase of triglycerides by 2-fold. Interestingly, the ratio of phosphatidylcholine to phosphatidylethanolamine was decreased, as previously suggested as a marker in the progression from steatosis to steatohepatitis.CONCLUSION: The thin layer chromatography analysis allows a reliable quantification of lipid classes and provides detailed insight into the lipogenic effect of p62.
基金funded by the Development and Reform Commission of Jilin Province (2010)
文摘[ Objective] To investigate the effects of concentrate/roughage ratios and nitrogen levels on in-vitro rumen fermentation of urea-treating corn stalk. [ Method] The concentrate/roughage ratios were 7： 3, 5：5 and 3： 7. The crude protein levels were 11% and 14%. The in-vitro culture time was 2, 4, 6 and 48 h. [ Result] The pH value of broth decreased significantly with the decline in the proportion of roughage （ P 〈 0.01 ） and with the increase in the crude protein levels （P 〈 0.05）. The ammonium nitrogen concentration and acetic acid/propionic acid ratio of the fermen- ted products decreased with the increase in the crude protein levels and proportion of concentrate in diet. The diet with concentrate/roughage ratio of 5：5 and crude protein level of 14% had significantly higher digestibility of dry matter and organic matter than other diets （ P 〈 0.05）. The digesti- bility of organic matter increased gradually with the increasing proportion of concentrate and crude protein level. With the increase in the proportion of concentrate, the microbial protein levels increased remarkably, while the acetic acid/propionic acid ratio declined. [ Conclusion] The concentrate/ roughage ratios and nitrogen levels affect rumen fermentation and microbial growth during in-vitro culture. However, the best supplementary feeding results of urea-treatinq corn stalks can be obtained when the concentrate/rouahaae ratio is below 5：5 and the crude protein level is 14%.
