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.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.
3.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.
4.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.
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.Determination of Five Kinds Halogenated Alkane Hydrocarbon in Drinking Water with Headspace Gas Chromatography
Shiping SUN ; Yan LI ; Jiangping DUAN
Journal of Environment and Health 1992;0(02):-
Objective To establish a test method of dichloromethane,1,1-dichloroethylene,1,2-dichloroethylene,1,2-dichloroethane in drinking water with headspace gas chromatography.Methods Halogenated alkane hydrocarbon in the water was extracted by headspace technique,then analyzed with DB-624 capillary column.In the same time,determined with GC by controlling the temperature.Retention time of the peaks was used for qualitative analysis,while external standard method was used for quantitative analysis.Results The linear ranges for dichloromethane,1,1-dichloroethylene,1,2-dichloroethylene,trans-1,2-dichloroethylene,1,2-dichloroethane were 0.85-168,0.07-12.1,0.40-77.8,0.53-119 and 1.2-265 ?g/L respectively,the determination limits were 0.83,0.07,0.40,0.53 and 1.10?g/L respectively,r≥0.999 5,the rate of recovery were 102.5%-113.8%,and RSDs were 5.5%-11.8%.Conclusion This method is simple,rapid and sensitive,can efficiently separate and accurately determine 5 kinds of halogenated alkane hydrocarbon in the water and only takes 5.2 minutes.
7.ANGIOGENESIS OF VASCULAR ENDOTHELIAL GROWTH FACTOR EVALUATED USING RAT SPONGE IMPLANT MODEL
Huijuan DUAN ; Yan LIU ; Tingsh YANG
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
To construct a repetitive, objective and consecutive system for evaluating the angiogenesis of vascular endothelial growth factor(VEGF) in vivo, a sterile polyvinyl alcohol sponge disc was implanted into the subcutaneous part of rat and five days later pCMV 4 VEGF 165 expressed plasmid and empty vector pCMV 4 were injected into the sponge, respectively. 7 and 11 days after injection, the tissue ingrowth in the implants and expression of VEGF gene in the serum, the tissues in the sponge and the liver were measured by RT polymerase chain reaction (RT PCR) .The pathological section of sponge was stained with hematoxylin and eosin (HE) and immunohistochemistry of Ⅷ factor specific for vascular endothelial cells. The tissue ingrowth in the implants was analysed by image analysis system and statistical analysis was done with Stata Software. The results indicated that the expression of VEGF gene was located at the injection site and promoted the tissue ingrowth in the implants. In brief, the angiogenesis of VEGF was successfully observed using the sponge implant model, which provides lays the theoretical foundation for local gene therapy of ischemic diseases.
9.Prognostic factor analysis of moderate-advanced hepatocellular carcinoma treated with sorafenib
Yan WANG ; Maoqiang WANG ; Feng DUAN
Journal of Interventional Radiology 2017;26(3):258-262
Objective To investigate the factors that may affect the prognosis of patients with moderate-advanced hepatocellular carcinoma (HCC) who are receiving sorafonib therapy.Methods The clinical data of 38 HCC patients,whose diagnoses were confirmed by pathology or clinical imaging and were in accord with the Barcelona Clinic Liver Cancer (BCLC) staging criteria,were retrospectively analyzed.Sorafenib 400 mg was orally given twice everyday,and according to modified RECIST criteria the tumor response was evaluated once every 4-6 weeks,the adverse events were recorded,and the drug toxicity was evaluated by grading standard 3.0 edition for common adverse reactions formulated by American National Cancer Institute.Results Among the 38 patients,one patient developed hepatic encephalopathy after took sorafenib for two weeks,and medication had to be stopped.Of other 37 patients,14 patients developed pancreatic atrophy in different degrees (positive group),the median progression free survival (mPFS) was 9.0 months,and the median overall survival (mOS) was 25.8 months;pancreatic atrophy didn't occurred in the remaining 23 patients (negative group),the mPFS was 3.3 months,and the mOS was 8.4 months.The differences in mPFS and in mOS between the two groups were statistically significant (P<0.01 and P=0.025respectively).The main adverse events included hand-foot skin reaction,diarrhea,fatigue and weight loss,which were relieved after symptomatic treatment,and no severe untoward effect occurred.Conclusion Certain correlation exists between pancreatic atrophy and the curative effect of sorafenib.For patients who develop severe diarrhea associated with pancreatic atrophy,a longer survival time as well as a longer stable state of disease can be expected.(J Intervent Radiol,2017,26:258-262)
10.Comparison of bioassay and HPLC for the determination of levofloxacin in plasma
Kaiying SI ; Jingli DUAN ; Baoxia YAN
Chinese Pharmaceutical Journal 1999;(1):43-
OBJECTIVE:The bioassay and HPLC methods were established and compared for the determination of levofloxacin(Lev) in human plasma.METHOD:In bioassay,medium Ⅱ and staphylococcus aureus were used.The HPLC method was chromatographed with a mobile phase of methanol-0.02mol/L potassium dihydrogen phosphate solution (3∶7),the sample was treated by adding 30% trichloroacetic acid to precipitate protein,and detected at 276mm.RESULTS:The average recovery of bioassay was within 95.90%~106.68%,the within-day and day-to-day precisions were below 6%.The average recovery of HPLC was within 97.24%~101.70%,the within day and day-to-day precisions were blow 5%.Lev tablets were given to 6 patients in two ways.Drug concentrations in plasma were determined by bioassay and HPLC assay,respectively,with no significant difference (P>0.05).CONCLUSION:The bioassay was simple,cheap,time saving,and reliable.The HPLC method wsa rapid,sensitive,accurate.The two methods may be selected to use in actual conditions.