AIM To study the possibility of matrix-assistedlaser desorption/ionization time of flight massspectrometry (MALDI--TOF MS) for controllingthe quality of recombinant proteins.METHODS By using MALDI--TOF MS, themoleCula...AIM To study the possibility of matrix-assistedlaser desorption/ionization time of flight massspectrometry (MALDI--TOF MS) for controllingthe quality of recombinant proteins.METHODS By using MALDI--TOF MS, themoleCular weights and purity of recombinantbioactive proteins were analyzed.RESULTS The molecular weights and puritywere obtained in nine recombinant bioactiveproteins, including interleukin 2, tumor necrosisfactor Q, granulocyte--macrophage colonystimulating factor, interferon aZb, interferon al,erythropoietin, calmodulin and its fragment, andneuronal nitric oxide synthase were obtained.MALDI--TOF MS was also used to assay specificproteins in the mixtures and to characterize theerythropoietin tryptic digests.CONCLUSION The results showed that MALDITOF MS can be employed for the effective qualitycontrol of recombinant proteins.展开更多
Metabolism of free fatty acids（FFAs） is related to several important physiological events and therefore their quantitaion in biological samples arouses extensive interest and efforts.Existing gas chromatography with...Metabolism of free fatty acids（FFAs） is related to several important physiological events and therefore their quantitaion in biological samples arouses extensive interest and efforts.Existing gas chromatography with flame ionization detector（GC-FID） methods for the analysis of FFAs normally require derivatization of them in order to lower boiling points.But this extra procedure tends to induce additional error and it is laborious and time-consuming.A derivatization-free method was therefore established in the present investigation to determine FFAs in human plasma by capillary（GC-FID）.After extraction of FFAs from plasma,a highly polar FFAP（free fatty acid in plasma） column was employed to directly quantitate FFAs concentration,free from derivatization reaction.All sample pretreatments were carried out at room temperature,improving recovery of short-chain FFAs.Heptadecanoic acid（C17：0） was employed as internal standard,and the proposed method was validated for recovery,precision,sensitivity,stabi-lity,and linearity.Validation data show that it is suitable for clinical study that has been applied to the evaluation of FFAs levels in plasma of diabetic nephropathy（DN） patients during the course of treatment.Forty-seven patients diagnosed with DN were admitted to the double-blind experiment.Control group（n=17） underwent solely basic treatment and the patients did not show significant change in FFAs concentration during six months of treatment.Experiment group（n=30） was supplied with traditional Chinese medicine besides basic treatment.After six months of medication,their plasma concentration of palmitic acid（C16：0）,stearic acid（C18：0） and oleic acid（C18：1n-9） decreased while linolenic acid（C18：3n-3） increased significantly（P〈0.05）.These four compounds could be served as biomar-kers in the evaluation of drug efficacy,and their quantitation in plasma may provide additional information for disease progression in DN patients.展开更多
1 Introduction As already known, complex prescription has been the outstanding feature of traditional Chinese medicine（TCM）. This fact therefore brings complicated formula with multicomponents from multimaterals, wh...1 Introduction As already known, complex prescription has been the outstanding feature of traditional Chinese medicine（TCM）. This fact therefore brings complicated formula with multicomponents from multimaterals, which produces some difficulties to detect the components and to control the quality. Shuanghuanglian preparations（SHLs）, including oral liquid, injection powder, capsule, and tablet, belong to a family of refined traditional Chinese patent medicine and a combined herbal remedy comprised of three herbs：展开更多
The authors focused their attention on the establishment of a mesenchymal stem cell（MSC） model for screening traditional Chinese medicines（TCMs） so as to investigate the effects of Shuanglong Formula（SLF） compon...The authors focused their attention on the establishment of a mesenchymal stem cell（MSC） model for screening traditional Chinese medicines（TCMs） so as to investigate the effects of Shuanglong Formula（SLF） components（Ginsenosides and salvianolic acids） and ingredients（ginsenoside Rb1 and salvianolic acid B） on cardiomyocyte differentiation from MSCs.The SLF components were analyzed and quantified by HPLC-TOF-MS.Cardiomyocyte differentiation was induced by culturing MSCs in the induction medium supplemented with SLF ingredients,SLF components,5-azacytidine（5-aza）,5-aza＋SLF ingredients and 5-aza＋SLF components,respectively,for up to 30 d,and evulated by the expression of Cardiac-specific myosin heavy chain（MHC） and troponin I（TnI） via immunofluoresent staining.Slow growth rate and changed morphology were observed during cardiomyocyte differentiation.After 20 d of induction,differentiating MSCs were positive for MHC and TnI staining.The effects of SLF components were better than those of SLF ingredients.Taken together,SLF can induce the differentiation of MSCs into cardiomyogenic cells in vitro,and MSCs can be used as a powerful tool for screening TCMs.展开更多
基金Supported by the National Natural Science Foundation of China,No.692350220.
文摘AIM To study the possibility of matrix-assistedlaser desorption/ionization time of flight massspectrometry (MALDI--TOF MS) for controllingthe quality of recombinant proteins.METHODS By using MALDI--TOF MS, themoleCular weights and purity of recombinantbioactive proteins were analyzed.RESULTS The molecular weights and puritywere obtained in nine recombinant bioactiveproteins, including interleukin 2, tumor necrosisfactor Q, granulocyte--macrophage colonystimulating factor, interferon aZb, interferon al,erythropoietin, calmodulin and its fragment, andneuronal nitric oxide synthase were obtained.MALDI--TOF MS was also used to assay specificproteins in the mixtures and to characterize theerythropoietin tryptic digests.CONCLUSION The results showed that MALDITOF MS can be employed for the effective qualitycontrol of recombinant proteins.
基金Supported by the National Basic Research Program of China(Nos.2007CB511903,2005CB523503)the International Cooperation Project of Ministry of Science and Technology of China(No.S2010GR0583)the National Natural Science Founda- tion of China(Nos.90709045,20805026)
文摘Metabolism of free fatty acids（FFAs） is related to several important physiological events and therefore their quantitaion in biological samples arouses extensive interest and efforts.Existing gas chromatography with flame ionization detector（GC-FID） methods for the analysis of FFAs normally require derivatization of them in order to lower boiling points.But this extra procedure tends to induce additional error and it is laborious and time-consuming.A derivatization-free method was therefore established in the present investigation to determine FFAs in human plasma by capillary（GC-FID）.After extraction of FFAs from plasma,a highly polar FFAP（free fatty acid in plasma） column was employed to directly quantitate FFAs concentration,free from derivatization reaction.All sample pretreatments were carried out at room temperature,improving recovery of short-chain FFAs.Heptadecanoic acid（C17：0） was employed as internal standard,and the proposed method was validated for recovery,precision,sensitivity,stabi-lity,and linearity.Validation data show that it is suitable for clinical study that has been applied to the evaluation of FFAs levels in plasma of diabetic nephropathy（DN） patients during the course of treatment.Forty-seven patients diagnosed with DN were admitted to the double-blind experiment.Control group（n=17） underwent solely basic treatment and the patients did not show significant change in FFAs concentration during six months of treatment.Experiment group（n=30） was supplied with traditional Chinese medicine besides basic treatment.After six months of medication,their plasma concentration of palmitic acid（C16：0）,stearic acid（C18：0） and oleic acid（C18：1n-9） decreased while linolenic acid（C18：3n-3） increased significantly（P〈0.05）.These four compounds could be served as biomar-kers in the evaluation of drug efficacy,and their quantitation in plasma may provide additional information for disease progression in DN patients.
基金Supported by the National S&T Supporting Project in the 11th Five-Year Period(No.2006BAI08B04-01).
文摘1 Introduction As already known, complex prescription has been the outstanding feature of traditional Chinese medicine（TCM）. This fact therefore brings complicated formula with multicomponents from multimaterals, which produces some difficulties to detect the components and to control the quality. Shuanghuanglian preparations（SHLs）, including oral liquid, injection powder, capsule, and tablet, belong to a family of refined traditional Chinese patent medicine and a combined herbal remedy comprised of three herbs：
基金Supported by the National Eleventh Five-Year Plan of China(No.2006BA108B04-01)the National Basic Research Program of China(No.2005CB523503)
文摘The authors focused their attention on the establishment of a mesenchymal stem cell（MSC） model for screening traditional Chinese medicines（TCMs） so as to investigate the effects of Shuanglong Formula（SLF） components（Ginsenosides and salvianolic acids） and ingredients（ginsenoside Rb1 and salvianolic acid B） on cardiomyocyte differentiation from MSCs.The SLF components were analyzed and quantified by HPLC-TOF-MS.Cardiomyocyte differentiation was induced by culturing MSCs in the induction medium supplemented with SLF ingredients,SLF components,5-azacytidine（5-aza）,5-aza＋SLF ingredients and 5-aza＋SLF components,respectively,for up to 30 d,and evulated by the expression of Cardiac-specific myosin heavy chain（MHC） and troponin I（TnI） via immunofluoresent staining.Slow growth rate and changed morphology were observed during cardiomyocyte differentiation.After 20 d of induction,differentiating MSCs were positive for MHC and TnI staining.The effects of SLF components were better than those of SLF ingredients.Taken together,SLF can induce the differentiation of MSCs into cardiomyogenic cells in vitro,and MSCs can be used as a powerful tool for screening TCMs.
