2.Determination of Dasatinib Concentration in Human Plasma by LC-MS/MS and Bioequivalence Study of 2 Kinds of Tablets
Huaiyou XU ; Chao SHU ; Feng SHAO ; Chunlei TAO
China Pharmacy 2016;27(8):1051-1054
OBJECTIVE:To establish a method for the determination of dasatinib concentration in human plasma and study the bioequivalence of 2 kinds of tablets. METHODS:In a randomized two-way crossover study,24 healthy male volunteers were divid-ed into two groups,and were administered respectively with test and reference preparations 100 mg under fasting and fed condi-tions. The plasma concentration of dasatinib was determined by LC-MS/MS. Using imatinib mesylate as internal standard,the deter-mination was performed on Welchrom C18 column with mobile phase consisted of acetonitrile-0.1% formic acid(70∶30,V/V). Posi-tive ion scanning was conducted under MRM mode,ESI,and ion pair for quantitative analysis were m/z 488.5→401.2 (dasatinib) and m/z 494.6→394.2(internal standard). DAS 3.2.8 software was used for data processing and variance analysis was adopted to in-vestigate the bioequivalence of 2 kinds of tablets. RESULTS:The linear rang of dasatinib was 1-300 ng/ml. The pharmacokinetic pa-rameters of test and reference preparations under fasting conditions were as follows:cmax were (165.599 ± 67.592) and (164.533 ± 77.960)ng/ml;tmax were(1.145±0.504)and(1.080±0.467)h;t1/2 were(5.080±2.262)and(3.771±1.596)h;AUC0-36 h were (550.487 ± 256.494) and (585.986 ± 324.885) ng·h/ml. The pharmacokinetic parameters of test and reference preparations under fed conditions were as follows:cmax were(163.058±47.533)and(165.440±53.012)ng/ml;tmax were(1.630±1.066)and(1.576± 0.530)h;t1/2 were(4.720±2.677)and(4.311±2.610)h;AUC0-36 h were(568.036±192.521)and(601.100±216.855)ng·h/ml. Relative bioavailability of AUC0-36 h under fasting and fed conditions were(100.2±7.5)% and(99.2±3.8)%. Analysis of variance showed there were no significant difference in the pharmacokinetic parameters of between 2 preparations and cyde(P>0.05),there was statistical significance among subjects(P<0.05). CONCLUSIONS:LC-MS/MS method can determine the concentration of da-satinib in human plasma rapidly;and the two preparations are bioequivalent.
3.Prospect and Progress on Dunaliella salina in the Area of Molecular Biology
Hong-Tao LIU ; Shu-Ying FENG ; Tao CHEN ; Le-Xun XUE ;
China Biotechnology 2006;0(10):-
Unicellular green alga,Dunaliella salina(D.salina),is a biflagellar alga without cell wall,which is a kind of very important eukaryotic microalga.In the previous study,the research of D.salina focus on the morphology,the mechanism of salt tolerance and ?-carotene,however,with the rapid development of microalgal biotechnology,a lot of work about D.salina was reported in recent years.In the area of molecular biology,the studies of D.salina mainly place emphasis on the cloning and analysis of important functional genes,regulatory sequences,and the expression of foreign genes using D.salina as host.The research advance in these aspects were reviewed.
4.Clinical performance of hepatitis B surface antigen quantification with high sensitivity chemiluminescent enzyme immunoassay technology
Chenli RAO ; Shu FENG ; Bin WEI ; Jingna AN ; Qixia CHEN ; Tingting WANG ; Chuanmin TAO ; Lanlan WANG
Chinese Journal of Laboratory Medicine 2015;(8):537-542
Objective To assess the clinic application of a new-style automated immunoassay system HISCL ( high sensitivity chemiluminescent enzyme immunoassay ) with high sensitive chemiluminescence substrate CDP-Star?, bind-free separate technology and filter conversion technology.Methods The performance verification test evaluated HISCL′s specification including reproducibility, functional sensitivity, linearity, accuracy, and sensitivity for HBsAg seroconversion.To evaluate the specificity , 1 007 HBsAg-negative specimens , 82 potentially interfering specimens were from conventional specimens in West China Hospital , Sichuan University between October and December of 2014.At the same time, with these results to determine the rate of the specimens that their results are in negative grey zone.259 HBsAg-positive specimens and 27 weakly-positive specimens were tested for the comparison between the HISCL and ECLIA ( electrochemiluminescence immunoassay ) HBsAg quantification.A linear regression model , Pearson′s correlation and Bland-Altman analysis were used as statistical methods.Results The between day and within-lot variable coefficient of two level samples are 1.55%, 2.02%, 0.34%, 1.34%separately.The functional sensitivity of the HISCL HBsAg assay reaches 0.007 IU /ml.There is a good linearity (y=3.262x+0.082,r=0.994; y=2 303.608x-33.006,r=0.999) within range of 0-2 300 IU/ml.HISCL obtain the accurate results for the CAP control material , the agreement rate is 100%.The sensitivity for the detection of HBsAg seroconversion is high.The specificity is 99.91%.The negative grey zone rate is 1.66%.The results of these two methods have good correlation and uniformity , r=0.995, LOA:-0.3 -0.19 log10 IU/ml.Conclusions HISCL HBsAg detection system shows good detection performance.Be of the function of both qualitative and quantitative detection.And the negative “grey zone”specimen rate is low.It is wholly capable of wide linearity and quick assays for quantifying serum HBsAg levels.HISCL could contribute to improve HBsAg quantitative detection process for clinical laboratory.
5.A comparative study of organ-system based integrated curriculum model and traditional teaching model in medical education
Fei TAN ; Baojun WAN ; Tao SHU ; Hui WEN ; Zhanchun FENG ; Jiagao LYU
Chinese Journal of Medical Education Research 2015;(5):468-472
Objective By comparing the organ system based medical integration teaching mode and traditional teaching mode to evaluate the effect of the integration of curriculum implemen-tation. Methods Through a questionnaire survey of 63 students who implement integrated curriculum and 183 students who are not implementing integrated curriculum as well as 76 teachers, we evaluate students' emotional, cognitive and motor skills, using SPSS statistical analysis, chi-square test. Results In the affective domain, most of the teachers thought the experimental class was superior to the com-mon one in learning interest, lifelong learning and independent learning, critical thinking, the contrast of the two classes were P=0.000, P=0.031 and P=0.001 respectively, all of significant difference. In the cognitive domain, the contrast of the two classes in memory, understanding and application were P=0.000, all of significant difference. The experimental class has high degree of recognition. In the psychomotor domain, more than 55.3%(42/76) of the teachers thought that the experimental class was superior to the common class. According to the students' questionnaire survey, the contrast of the two classes in clinical operation skills, Interpersonal communication skills were P=0.077, P=0.031 respec-tively. There was no statistically significant difference. Conclusions The integrated curriculum teach-ing model can greatly promote students' abilities in most areas such as interest in learning, lifelong learning awareness, clinical thinking ability; interpersonal communication skills etc. In some respects the differences remains to be further studied such as memory of knowledge, systematic knowledge, and clinical operation skills, etc.
6.Preliminary Study on Transformation of Human canstatin Gene to the Novel Bioreactor——Dunaliella salina
Shu-Ying FENG ; Hui-Hui GU ; Hong-Tao LIU ; Le-Xun XUE ;
China Biotechnology 2006;0(06):-
The human canstatin cDNA was amplified by RT-PCR and then directionally cloned into pU? expression vector. The recombinant pU?-Can vector was connected with the screening marker (bar box), to construct a eukaryotic expression vector called pU?-Can-Bar. This expression vector was introduced into the D.salina by glass beads method. The screening culture of transformants of D.salina was performed in solid media containing 5 ?g/ml PPT, and the analyses of transformants were carried out through PCR and Southern blot. PCR results revealed that specific 700 bp products were detected in the different transformants of D.salina but not in negative control. Southern blot analysis further demonstrated that human canstatin gene was integrated into the D.salina genome. Moreover, the results of genetic stability analyses of transformants demonstrated that canstatin gene was stably inherited in the D.salina transformants. The successful preparation of the D.salina transformants will provide the experimentation evidence for producing canstatin protein cosmically by using the D.salina bioreactor and give a better prophase work basis for clinic application of canstatin protein early.
7.Anti-fatigue Effect of Cynomorium Songaricum Flavone on Old Rats
Fa-rong YU ; Shu-tao FENG ; Ming-ren XIE ; Xiuzhen LIAN
Chinese Journal of Rehabilitation Theory and Practice 2008;14(12):1141-1142
Objective To study the anti-fatigue effect of Cynomorium songaricum flavone(CSF)on old rats.Methods50 old male Wistar rats were randomly divided into the control group(no swimming,no drugs),single swimming group(swimming,given saline by gavage),and three swimming plus CSF groups(swimming,given CSF 20 g/kg weight,10 g/kg weight and 5 g/kg weight respectively),total five groups with 10 animals in each group.Animals swam with free swimming in endless cycles water 30 minutes once every day for 10 days.The level of monoamine oxidase(MAO) and glutathione peroxidase(GSH-px) in blood of animals were examined by MAO kit,GSH-px kit and ultraviolet spectrophotometer at 11th day.ResultsCompared with single swimming group,the body weights of swimming plus CSF groups increased significantly by 12.9%,80.6% and 200% respectively(P<0.01);swimming times before sinking increased by 19.4%,29% and 48.4%;sinking times of each rat(time/10 min) decreased by 17.8%,43.2% and 72.9%;total swimming times increased by 2.8%,7% and 29.6% respectively(P<0.01);MAO decreased by 60%,69.8% and 80.5%,GSH-px increased by 465.3%,563.8% and 635% respectively(P<0.01).ConclusionCSF shows anti-oxidative and anti-fatigue properties and can be given as prophylactic/therapeutic supplements for increasing antioxidant enzyme activities and preventing lipid peroxidation during strenuous exercise.
8.Effects of trimetazidine on myocardial autophagy in rats with heart failure after myocardial infarction
Shu-Na TAN ; Jin-Ping FENG ; Chao FENG ; Cui-Yan TIAN ; Shu-Tao CHEN
Tianjin Medical Journal 2018;46(2):152-156,108
Objective To observe changes of left ventricular function and the level of autophagy after treatment with trimetazine in rats with heart failure after myocardial infarction(MI). Methods Twenty healthy male Wistar rats with the ligation of the proximal part of the left descending branch were randomly and equally allocated into two groups,model group (M group) and trimetazine group (Q group). A sham group (S group) was made up by 10 sham-operated rats. Rats of trimetazine group were given trimetazine(15 mg/kg)once a day for 4 weeks.Then left ventricular function was measured by echocardiography,and hemodynamics was evaluated by Millar pressure-volume system.Serum levels of NT-proBNP and hs-TnT were tested by ELISA.Pathological changes and fibrosis of myocardium were observed by HE and Masson staining.The myocardial apoptosis level was observed by TUNEL, and expressions of autophagy related protein and gene in myocardial tissue were detected by Western blot assay and RT-PCR. Results Trimetazine treatment significantly improved left ventricular dilatation and dysfunction in rats with myocardial failure. Trimetazine treatment also significantly improved pressure overload and the compliance decrease of left ventricular in rats with heart failure after myocardial infarction. Trimetazidine reduced the edema,necrosis and myocardial fibrosis of cardiac myocytes in rats with heart failure.The results from ELISA showed that serum levels of NT-proBNP and hs-TnT were significantly lower in the trimetazine group than those of model group.Compared with model group,the cardiomyocyte apoptosis decreased significantly in the trimetazine group.The results from Western blot assay and RT-PCR showed that autophagic flow of myocardium was increased remarkably in the trimetazine group than that of model group. Conclusion Autophagy has a protective effect on myocardial cells. Trimetazine can improve cardiac function through up-regulation of autophagy in cardiomyocytes in MI rats with heart failure.
9.Isolation, identification, culture and bionomics of skeletal muscles satellite cells from green fluorescent protein transgenic mouse in vitro.
Feng-he ZHANG ; Feng-cai WEI ; Ping HUANG ; Shu-yang SUN ; Ke-tao WANG
West China Journal of Stomatology 2007;25(2):188-191
OBJECTIVETo investigate the green fluorescent protein (GFP) expression and the bionomics of skeletal muscles satellite cells (SMSCs) in vitro in GFP transgenic mouse.
METHODSThe newborn transgenic mice were acquired to separate skeletal muscles satellite cells with enzyme digestion method. Cells were cultured and subcultured in vitro. Morphological observation, growth curve were investigated to evaluate the proliferation and differentiation characteristics of skeletal muscles satellite cells, fluorescence microscope was used to observe the GFP expression. The cells were identified by immunocytochemical stain. In the basis of identification of anti-sarcometric actin anti-body, the combination of anti-desmin antibody and DAPI (4, 6-diamidino-2-phenylindole) were used to detect the purification of skeletal muscles satellite cells.
RESULTSImmunocytofluorescence suggested the good retain of GFP fluorescence in skeletal muscles satellite cells. The cells showed strong proliferative ability and they were positive with immunocytochemical stain of anti-sarcometric actin antibody and anti-desmin antibody. The combination of anti-desmin and DAPI stain can be used to determine the purification of SMSCs.
CONCLUSIONSkeletal muscles satellite cells cultured in vitro showed strong proliferation and differentiation ability. They are fit to construct the cell bank of tissure engineering and to be a useful tool to explore cells fate after transplantation since these cells retain the expression of GFP.
Actins ; Animals ; Autoantibodies ; Cell Differentiation ; Cells, Cultured ; Desmin ; Green Fluorescent Proteins ; In Vitro Techniques ; Mice ; Mice, Transgenic ; Muscle, Skeletal ; Satellite Cells, Skeletal Muscle
10.Patients' quality of life after laparoscopic or open cholecystectomy.
Li CHEN ; Si-feng TAO ; Yuan XU ; Fu FANG ; Shu-you PENG
Journal of Zhejiang University. Science. B 2005;6(7):678-681
OBJECTIVEThis study was aimed at evaluating and comparing the quality of life in patients who underwent laparoscopic and open cholecystectomy for chronic cholecystolithiasis.
METHODSThe study included 25 patients with laparoscopic cholecystectomy (LC group) and 26 with open cholecystectomy (OC group). The quality of life was measured with the Gastrointestinal Quality of Life Index (GLQI) preoperatively, thereafter regularly at 2, 5, 10 and 16 weeks after the operation.
RESULTSThe mean preoperative overall GLQI scores were 112.5 and 110.3 in LC and OC group respectively (P>0.05). In the LC group, the mean overall GLQI score reduced slightly to 110.0 two weeks after the operation (P>0.05). The LC group showed significant improvement in overall score and in the aspects of symptomatology, emotional and physiological status from 5 to 16 weeks postoperatively. In the OC group, the GLQI score reduced to 102.0 two weeks after surgery (P<0.05). Significant reductions were shown in the aspects of symptomatology, physiological and social status. The GLQI scores returned to the preoperative level of 115.6 ten weeks after the operation (P>0.05). The patients experienced significant improvements of GLQI sixteen weeks after OC operation (P<0.01~0.05). Within the 10 postoperative weeks, the LC group had significantly higher GLQI scores than the OC group (P<0.05).
CONCLUSIONSLC can improve the quality of life postoperatively better and more rapidly than OC. The assessment of quality of life assessment is a valid method for measuring the effects of surgical treatment.
Adult ; China ; epidemiology ; Cholecystectomy ; statistics & numerical data ; Cholecystolithiasis ; epidemiology ; surgery ; Comorbidity ; Female ; Health Status ; Humans ; Laparoscopy ; statistics & numerical data ; Male ; Middle Aged ; Pain, Postoperative ; epidemiology ; Patient Satisfaction ; Postcholecystectomy Syndrome ; epidemiology ; Quality Assurance, Health Care ; methods ; Quality of Life ; Treatment Outcome