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Objective@#To establish a triple-color pseudovirion-based neutralization assay (PBNA) and evaluate its capability of detecting immunogenicity of the sera generated by the immunization of HPV 9-valent vaccine.@*Methods@#HPV pseudovirus (PsVs) 6/11/16/18/31/33/45/52/58 with the encapsidated fluorescence expressing red fluorescent plasmid N31-MCHREEY, green fluorescent N31-EGFP or blue fluorescent N31-mTagBFP were generated. The concentration of HPV PsVs and the infection titers of HPV PsVs were detected by double-antibody sandwich ELISA and TCID50, respectively. The single- and triple color HPV 16/33/45 PsVs were used to detect the neutralization titers of mice sera immunized with HPV 9-valent vaccine and confirmed the accuracy and specificity of the triple-color PBNAs. Then, the single- and triple color HPV 6/11/18/31/33/45/52/58 PsVs were employed to detect the neutralization titers of cynomolgus macaques sera immunized with HPV 9-valent vaccine and determined whether the triple-color PBNAs could be applied to evaluate the immunogenicity of the sera generated by the immunization of HPV9-valent vaccine.@*Results@#The concentration of HPV16 PsVs encapsulating green, red or blue fluorescent plasmid was 5.0 to 6.0 μg/ml and HPV6/11/18/31/33/45/52/59 triple-color HPV PsVs was about 1.0 to 3.0 μg/ml. 9 types HPV PsVs containing EGFP, Mcherry or mTagBFP reporter plasmid were obtained and the concentration can meet the need of neutralization detection. 9 types single-color fluorescent HPV PsVs had similar infectivity against 293FT cells with the infection titer values between 1×104 and 1×105. The results of PBNAs showed that there was no significant difference in the anti-HPV neutralization titers of mice sera induced by HPV 9-valent vaccine between single-color and triple-color HPV16/33/45 PsVs (P>0.05). Similarly, there was also no significant difference in the anti-HPV neutralization titers of cynomolgus macaques sera induced by HPV 9-valent vaccine between single-color and triple-color HPV6/11/18/31/33/45/52/58 PsVs (P>0.05).@*Conclusion@#We successfully established the triple-color PBNAs and verified the accuracy and specificity of triple-color PBNAs consistent with single-color PBNAs. The triple-color PBNAs can be applied to evaluate the immunogenicity of HPV 9-valent vaccine's immune serum.
ABSTRACT
Objective To compare the application effect between problem-based learning (PBL) and traditional teaching in cardiovascular intervention . Methods 39 training physicians were divided randomly into the PBL group (n=20) and control group (n=19). The control group was trained with the tradi-tional teaching method while PBL group used PBL seven step method, namely they were trained through the process of clarifying unfamiliar terms—defining the problem—brainstorming—restructuring problem—defining learning goals—collecting information, personal learning, information sharing, and group discus-sion. After the end of the training, the two groups were tested by using the unified test questions and skills test, and the questionnaire survey of teaching satisfaction. SPSS 18.0 was used to do line t test or chi square test to the data of both groups. Results PBL group training physicians' cardiovascular intervention oper-ation [(30.07±1.67) vs. (28.54±1.98), P=0.036], their comprehensive analysis of clinical cases, [(34.47± 1.77) vs. (32.08 ±1.80), P=0.002], and the total score [(86.47 ±2.75) vs. (82.23 ±3.63), P=0.002], were better than the control group, and the difference was statistically significant. The survey results showed that the PBL group's evaluation on how the teaching methods stimulate the training physicians' interest in learning, enhance their ability of independent thinking and cultivate their teamwork ability, improve their language expression and clinical thinking and other aspects was higher than the control group (P Conclusion Compared with the traditional teaching, the application effect of PBL in the training of car-diovascular intervention can better exert training physicians' subjective initiative and improve the teaching effect.
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To observe Antiarrhythmic effects of L-dicentrine．Methods using experimented model of arrhythmials．Results: L-dicentrine iv l0 mg/kg could convert BacRinduccd arrhythmia into sinus rhythm in rats,and could significantly increase the tolerant dose of aconine to produce Ventricular fibrillation (VF) and Cardiac ［rrest (ca) in rats． The drug could also decrease the incidence of VF and CP by cardiac rats and by chloroform in mice． L-dicelltrine had protective effects on ouabain ind uced Arrhythmias in guined pigs and on adrenalice induced arrhythmias in rabbit s ． The drug might also inhibit the arrhymias induced by stimulation ofhypothaplu s． Conclusion: L-dicenirine have significant antiarrhythmic properties．