1.Establishment and verification of purity detection method for receptor-binding domain dimer in SARS-CoV-2 recombinant protein vaccine
Chinese Journal of Biologicals 2024;37(10):1239-1244+1250
Objective To establish and verify a bioanalyzer(Agilent 2100) method for the determination of the receptor-binding domain(RBD) dimer purity in SARS-CoV-2 recombinant protein vaccine.Methods The protein in the SARS-CoV-2recombinant vaccine was analyzed and then concentrated by ultrafiltration, and the purity of fully denatured and partially denatured(disulfide bond retention) proteins was detected by bioanalyzer(Agilent 2100). The established method was verified for the applicability, specificity, reproducibility, precision and accuracy, and used to detect four batches of samples from manufacturer A.Results In the system applicability test, eight peaks were clearly distinguished from the baseline. The blank solvent had no interference at the peak position of the target component, indicating the good specificity. Under the fully denatured and partially denatured conditions, the RSDs of 12 repeated test results by two experimenters were both less than 1%,showing good repeatability and precision. The RSDs of the proportion of the main peak(1+2) and the main peak at three different protein concentrations(50%, 80% and 100%) were 0. 68% and 0. 31%, respectively, and the accuracy was good.Among them, the results of the 10% protein concentration sample were unstable. The purity of RBD dimer in four batches was 70. 5%, 70. 2%, 73. 2% and 69. 6%, respectively.Conclusion The established method has good specificity, precision and accuracy, and can be used for the detection of protein purity in recombinant protein vaccines.
2.Study on Rank—Frequency Distribution of Scientific Research Papers by famous teacher teaching in biomedicine domain
Chinese Journal of Medical Science Research Management 2012;25(3):206-209
Study on Rank—Frequency Distribution of Scientific Research Papers by Chinese University famous teacher teaching in biomedicine domain from 2003 to 2009.Whose quantitative indicators of scientific papers were negative logarithmic distribution,except provincial-level teaching famous teachers' SCI papers were negative exponential distribution. The qualitative index were negative logarithmic,except state-level teaching famous teachers' SCI paper yearly average paper cited were linear distribution and provincial-level were negative exponent distribution.The magnitude of concentrations was between 0.257 and 0.728,which deviates from the Price's law (0.50). It indicates that the Rank-frequency distribution model can be applied to the teaching famous teacher.The internal gradation instrument in the top scientists is different from in science community.Scientist's gradation in the science community depends on whether the objective of scientific paper evaluation comes true.The state-level teaching famous teacher scientific paper difference is greater than the provincial-level; SCI paper difference is bigger than the CSCD paper.
3.Simultaneous Determination of Phenylpropanoids in Drug Pairs of Chuanxiong Rhizoma and Angelicae Sinensis by HPLC
China Pharmacist 2015;18(10):1718-1720
Objective: To determine the content of phenylpropanoids in the drug pairs of Chuanxiong Rhizoma and Angelicae Sinensis. Methods:An ISC-HPLC method was used to determine the content of ferulic acid, protocatechuic acid and chlorogenic acid. Results:The content of ferulic acid, protocatechuic acid and chlorogenic acid in the drug pairs of Chuanxiong Rhizoma and Angelicae Sinensis all had good linear relationship with the peak area within the range of 0. 90-14. 40,1. 25-20. 00, and 1. 00-16. 00μg·ml-1 , respectively, and the average recovery was 97. 16%, 94. 98% and 98. 14%, respectively (n=9). Conclusion:The HPLC method is simple, rapid, accurate and reproducible, which can be used for the quality control of the drug pairs of Chuanxiong Rhizoma and An-gelicae Sinensis.
4.Measurement of immunophenotype and cytotoxicity of CIK against tumor cells in vitro by FCM
Journal of Jilin University(Medicine Edition) 2006;0(04):-
Objective To investigate the immunophenotype and the cytotoxicity of cytokine-induced killer(CIK) against tumor cells in vitro.Methods Lymphocytes cells were isolated freshly from peripheral blood of healthy donors by Ficoll-Hypaque density centrifugation,and the cells obstained were induced by IFN?,IL-2 and CD3McAb.Phenotypes and cytotoxicity of CIK were analysed by FACS.Target cells were differentiated from effect cells by CFSE dying.The mortality of Target cells were determined by FACS.Results The maximum proliferation of CIK reached at the 22nd day.The phenotypes of CD3,CD11a,CD54,HLA-DR were expressed highly;CD25,CD28,CD69,FasL were expressed moderately on CIK.The expression of CD16 was not increased.CIK possessed the cytotoxicity against tumor cells of K562,HL-60,Hela,SMMC7721 and A375.Conclusion IFN?,CD3McAb,IL-2 can induce peripheral lymphocytes to produce CIK which own strongly proliferation and exert highly efficient cytotoxic effects on tumor cells.
5.Effects of mifepristone on fibronectin in human villus and deciduas
Ling, GAO ; Yan, LIU ; Duan-duan, LA
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(6):722-724
Objective To investigate the effects of mifepristone on the expression of fibronectin (FN) in human villus and deciduas after termination of pregnancy. Methods Women with choice of termination of pregnancy(< 49 d) were divided into medical abortion complete group (n=15), medical abortion incomplete group(n=15) and control group (with dilatation and curettage, n=15). The villus and deciduas were collected at the third day of medical abortion or at the day of dilatation and curettage. Immunohistochemical technique was used to examine the expression of FN in villus and deciduas of these three groups. Results The positive expression rates of FN in villus and deciduas of control group were significantly higher than those of two medical abortion groups(P < 0.017), and the positive expression rates of FN of medical abortion incomplete group were significantly higher than those of medical abortion complete group (P < 0. 017). Conclusion Mifepristone can decrease the expression of FN in human villus and deciduas and interfere the condition of embryonic growth, exerting the anti-gestational effect at extracellular matrix level.
6.Progress in treatment of childhood Hodgkin lymphoma.
Yan-Long DUAN ; Yong-Hong ZHANG
Chinese Journal of Hematology 2011;32(8):567-571
Child
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Hodgkin Disease
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therapy
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Humans
7.Reform of pharmacognosy teaching materials of clinical pharmacy
Jianhui WANG ; Jingyu DUAN ; Yan LI
Chinese Journal of Medical Education Research 2006;0(08):-
In order to enhance the students’knowledge of clinical medicine and lay the foundation for using clinical medicine,we adjusted the content and compiled pharmacognosy teaching materials,aiming at its professional features of clinical pharmacy of Xuzhou Medical College.The pharmacognosy teaching material has been applied in several rounds.Feedback from students suggested that using this teaching material was well-targeted,clear and practical.
8.Study on the medical services monitoring under the concept of big data
Yan ZHANG ; Lei DUAN ; Liang ZHANG
Chinese Journal of Health Policy 2017;10(7):71-74
Medical service utilization monitoring of county areas residents is one of the important contents of health policy management and research.Traditional service utilization monitoring, such as National Health Services Survey (NHSS), residents healthcare treatment (clinical) data from medical insurance, had problems of high cost, large deviation, slowly updating monitoring indicators, out of time in health management decisions.The data convergence and re-architecture of multi-source data provide a new idea for the monitoring of residents'' service utilization.Different linking data-sources would come true with the tools of Microsoft Access database administration and Excel programming techniques.At the same time, a new database would be accessed with the indices of residents'' annual medical service utilization, preference in medical services, unconscionable service utilization, and lead to the promotion of medical service utilization monitoring research development.
9.Effect of mmu-miR-30b on mouse FoxO3 mRNA expression
Zhiqing DUAN ; Lu LI ; Yan LI
Chinese Journal of Microbiology and Immunology 2017;37(8):580-585
Objective To analyze the direct interaction between mmu-miR-30b and mouse FoxO3 (mFoxO3) mRNA.Methods Three target gene fragments, which were respectively 402, 123 and 299 bp in length, were amplified from mouse cDNA by PCR using specific primers and site-direct mutant primers.A complete mutant fragment was obtained by joining together the 123 and 299 bp fragments.Recombinant plasmids, pmirGLO-mFoxO3 and pmirGLO-mFoxO3 mt, were constructed through inserting wild and mutant fragments of mFoxO3 into pmirGLO vector, respectively.HEK 293T cells were respectively co-transfected with the constructed recombinant plasmids and mmu-miR-30b/mmu-miR-30b inhibitor.Dual-luciferase reporter assay system was used to determine the Firefly-Renilla luciferase activity in different groups.Western blot assay was performed to evaluate the regulatory effect of mmu-miR-30b on mFoxO3 expression.ResultsRestriction enzyme analysis and gene sequencing showed that the two recombinant plasmids, pmirGLO-mFoxO3 and pmirGLO-mFoxO3 mt, were constructed successfully.The activity of Firefly-Renilla luciferase in HEK 293T cells transfected with pmirGLO, pmirGLO+mmu-miR-30b, pmirGLO+mmu-miR-30b inhibitor, pmirGLO-mFoxO3+mmu-miR-30b, pmirGLO-mFoxO3+mmu-miR-30b+mmu-miR-30b inhibitor, or pmirGLO-mFoxO3 mt+mmu-miR-30b was 13.18±0.97, 14.35±0.99, 12.46±1.20, 9.55±1.11, 13.71±0.89 and 10.99±0.92, respectively.Compared with pmirGLO+mmu-miR-30b group, the luciferase activity was significantly decreased in pmirGLO-mFoxO3+mmu-miR-30b group (P<0.05), but showed no significant change in pmirGLO-mFoxO3 mt+mmu-miR-30b group (P>0.05).In addition, the suppressed Firefly-Renilla luciferase activity in pmirGLO-mFoxO3+mmu-miR-30b group was restored by mmu-miR-30b inhibitor treatment (P<0.05).Enhancing the expression of mmu-miR-30b could markedly inhibit the expression of mFoxO3 at protien level (P<0.05), and that could be significantly attenuated by mmu-miR-30b inhibitor treatmeat (P<0.05).Conclusion mFoxO3 mRNA is a novel target gene of mmu-miR-30b.There is a direct interaction between mmu-miR-30b and mFoxO3 mRNA.
10.Effects of Jinzhuyalong Ershiwuwei coral capsule on PLA2 in degenarated cervical intervertebral discs
Yan JIAO ; Hong DUAN ; Xiaoyang LUO
International Journal of Traditional Chinese Medicine 2013;(7):610-612
Objective To study the anti-inflammation mechanism of Jinzhuyalong Ershiwuwei coral capsule on cervical spondylosis by investigating Jinzhuyalong Ershiwuwei coral capsule on PLA2 in degenerated cervical intervertebral discs.Methods 60SD rats (SPF) were divided into a control group (15),a medicine model control group (15),a medicine modle 1 group (15),and a medicine model 2 group (15) by random lottery; Medicine model 1,2 groups were given Jinzhuyalong Ershiwuwei coral capsule powder 0.33 × 5 g/kg 0.66×5 g/kg,and added 1 ml distilled water dilution lavage,l/d,90 d,The unbalance of power cervical spondylosis animal models were chosen and C4~5 neck intervertebral discs were taken after 90 days.Millipore colorimetry methods were used to measure PLA2 activity in C4~5 cervical intervertebral discs.Results The PLA2 activity of the cervical intervertebral disc (12.37 ± 4.21) IU/mg · min-1 in the model group was significantly higher than the control group(2.56± 1.19)IU/mg· min-1 (P<0.05).The cervical intervertebral disc PLA2 activity of medicine model group 1 (8.56± 2.13)IU/mg · min-1 and medicine model group 2 (5.58 ±2.39)IU/mg· min-1 was lower than the model control group(12.37±4.21)IU/mg· min-1,with significant differences (both P<0.01).Conclusion Jinzhuyalong Ershiwuwei coral capsule could down regulate PLA2 activity in degenerated cervical intervertebral discs,and also decrease the production of some inflammaroty mediators.