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Objective:To establish the protein content determination method respectively by Folin-Ciocalteu method and Coomass-ie brilliant blue binding method for mannatide oral solution and compare the results. Methods:The limit of detection, linearity, accu-racy, repeatability, recovery and content of Folin-Ciocalteu method and Coomassie brilliant blue binding method were investigated. Re-sults:As for Folin-Ciocalteu method, the limit of detection was 0. 2 μg, the range of linearity was 0-112. 0 μg(r=0. 9990), the av-erage recovery was 105. 2%(RSD=1. 9%), the RSD of accuracy was less than 1%, and the content result of three batches of samples was 47. 45, 58. 34 and 40. 99 μg·ml-1, respectively. As for Coomassie brilliant blue binding method, the limit of detection was 0. 8 μg, the range of linearity was 0-99. 6 μg(r=0. 9980), the average recovery was 102. 0%(RSD=2. 7%), the RSD of accuracy was less than 1%, and the protein content of samples was not detected out by the method. Conclusion:Folin-Ciocalteu method can be used to determine the protein content in mannatide oral solution reliably and efficiently.
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Objective:To establish the protein content determination method respectively by Folin-Ciocalteu method and Coomass-ie brilliant blue binding method for mannatide oral solution and compare the results. Methods:The limit of detection, linearity, accu-racy, repeatability, recovery and content of Folin-Ciocalteu method and Coomassie brilliant blue binding method were investigated. Re-sults:As for Folin-Ciocalteu method, the limit of detection was 0. 2 μg, the range of linearity was 0-112. 0 μg(r=0. 9990), the av-erage recovery was 105. 2%(RSD=1. 9%), the RSD of accuracy was less than 1%, and the content result of three batches of samples was 47. 45, 58. 34 and 40. 99 μg·ml-1, respectively. As for Coomassie brilliant blue binding method, the limit of detection was 0. 8 μg, the range of linearity was 0-99. 6 μg(r=0. 9980), the average recovery was 102. 0%(RSD=2. 7%), the RSD of accuracy was less than 1%, and the protein content of samples was not detected out by the method. Conclusion:Folin-Ciocalteu method can be used to determine the protein content in mannatide oral solution reliably and efficiently.
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Objective:To evaluate the quality status of lithium carbonate tablets and lithium carbonate sustained-release tablets. Methods:The samples were examined in accordance with the statutory standard,and the exploratory studies were carried out. The results were statistically analyzed. Results:In accordance with the statutory standard,among 120 batches of samples, only one was unqualified in dissolution,and the others were qualified. The qualified rate was 99. 2% . Conclusion:The quality of the most products meets the current standard and the quality evaluation standard needs to be improved.
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Objective:To establish a method for the determination of chloride and sulfate in lithium carbonate crude drug by sup-pressed ion chromatography. Methods:The analysis column and the guard column was Thermo IonPac AS11(250 mm × 4 mm) and IonPac AG 11(50 mm ×4 mm), respectively. Potassium hydroxide solution(10 mmol·L-1) generated by an eluent generator(EG) was used as the eluent at a flow rate of 1. 0 ml·min-1 . And the temperature of column and conductivity cell was 30℃ and 35℃, re-spectively. The suppressor electric current was 36 mA, and the injection volume was 10 μl. Results:The linear range of chloride ion and sulfate ion was 1. 0-20μg·ml-1 with r of 0. 995 1 and 0. 997 3,respectively, and the recovery was 106. 6%(RSD=2. 6%,n=6) and 100. 3%(RSD=1. 9%,n=6), respectively. Conclusion:The method is accurate and reliable, and can be used in the determina-tion of chloride and sulfate in lithium carbonate crude drug.
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Objective:To explore the role of TNFSF14 and its receptors LTβR and HVEM in the pathogenesis of virus hepatitis.Methods:Marine fulminant viral hepatitis model was established by infecting mice with MHV-3.Liver tissue destruction in LIGHT KO and WT mice were analyzed by HE staining and ALT levels in serum by automatic biochemical analyzer .The mRNA levels of HVEM and LTβR in the liver and spleen tissues in the indicated time points ( 0 h, 12 h, 24 h, 48 h, 72 h ) were detected by quantitative-PCR.The expression of HVEM and LTβR on PBMC in patients with severe hepatitis were measured by flow cytometry.Results:In the MHV-3-induced murine fulminant hepatitis model ,liver injury in LIGHT KO mice was obviously decreased than that of WT mice,and ALT levels was also significantly lower than that of WT mice (P<0.01).The mRNA of HVEM and LTβR in the spleen were increased significantly after 48 h postinfection with MHV-3 ( P<0.05 );The level of LTβR mRNA in liver was significantly up-regulated in 12 h postinfection with MHV-3(P<0.01).Compared to healthy volunteers,the expression of both HVEM and LTβR on PBMC in patients with severe hepatitis was remarkably enhanced .Conclusion: TNFSF14 and its receptors LTβR and HVEM play a critical role in the pathogenesis of viral fulminant hepatitis .
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Objective:To investigate the effect of TNF-αand IFN-γon NIT-1 cells apoptosis and the apoptosis mechan-ism.Methods:NIT-1 cells were exposed to a combination of TNF-αand IFN-γtreatment.The viability of NIT-1 cells was assessed via MTT assay.The morphological changes of the cells and nuclei were observed under the inverted or confocal laser scanning microscope with Hoechst 33258 staining.The activation of Caspase-8,-3 and PARP was detected by Western blot.Results:TNF-αand IFN-γsig-nificantly inhibited NIT-1 cells viability, promoted cells apoptosis, induced the activation of Caspase-8,-3 and PARP cleavage.Conclusion:TNF-αcombined with IFN-γtreatment induced the apoptosis of NIT-1 cells through death receptor pathway.
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Ph.D candidate education is the highest level of higher education. Training model of Ph.D candidate in Medical College of Georgia and University of Manitoba)has vivid characters compared with that in China,which is reflected by the training objective,qualification of students and tutors,culti-vating procedures and admission requirements for graduation. This kind of cultivating model performs stringent selection and can gradually pick out persons who are really fit for the scientific research. Ph.D candidate quality in the two universities is guaranteed by systemic and deep courses learning,immediate update of knowledge and strict evaluation system. The goal of this article is to provide experience and ref-erence for improving the education quality of medical Ph.D candidates in China.
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Objective To obtain Chinese hamsterovary (CHO) cell line expressing human decay accelerating activity (hDAF) stably and to observe the protective effect of hDAF on heterologous cells under the circumstance of complement activation. Methods The eukaryotic expression vector DAF pcDNA3.1 was constructed and then transfected into CHO cells by lipofection. Monoclones of cells expressing hDAF stably were screened by the method of limiting dilution. hDAF expression was detected by flow cytometry. The decay accelerating activity of hDAF was determined by assay of C3 deposition and 51Cr release. Results The expression vector DAF pcDNA3.1 was successfully constructed, and monoclones of cells expressing hDAF were obtained. CHO cells expressing hDAF could decrease C3 deposition and attenuate the killing effect of activation of the complement system. Conclusion We have obtained CHO cell clones expressing hDAF stably, which is helpful for the further studies of the relationship of the structure with the functions of hDAF.
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Objective To investigate the activation of human T cells with superantigen SEA. Methods T cells proliferation, IL-2 production, DNA synthesis, cell phenotype and apoptosis induced by the SEA in the first or restimulation were detected. Results It was shown that 100 ng/mL was the optimal activation concentration, IL-2 production arrived at top level in the third day, SEA activated CD4+ and CD8+T with the same degree, T cells start apoptosis in response to SEA restimulation within 24 hours and apoptosis disappeared through addition of rhIL-2. Conclusions There were correlation between activation and SEA concentration or stimulation period； SEA activated both CD4+ and CD8+T cells without change the cell phenotype.
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Objective:To explore the relationship between transcription factor AP 1 and NF kappaB and IL 2 decrease in anergic T cells.Methods:Nuclear proteins of activated T cells and anergic T cells were extracted and then the expression of AP 1 and NF kappaB were determinated by gel electrophorotic mobility shift assay.Results:Compared to activated T cells,anergic T cells expressed defective AP 1 transcription factor;there were three forms NF kappaB complex in both activated T cells and agergic T cells,but NF kappaB transcription activity was higher in anergic T cells than that in activated T cells.Conclusion:It has been demonstrated that these alteration of transcription factors have been likely to be responsible for repression of IL 2 in anergic T cells.