Objective To develop an indirect ELISA method for quantitative detection of antibodies against pertussis toxin(PT), filamentous hemagglutinin(FHA) and pertussis adhesin(PRN) in serum of mice, and verify and preliminarily apply the method in order to provide reference for the optimization of production process of pertussis vaccine.Methods Enzyme-labeled plates were coated with PT, FHA and PRN antigens respectively, and the pertussis anti-mouse serum standard and mouse serum samples were added. Using goat anti-mouse IgG labeled with HRP as the enzyme-labeled secondary antibody,the contents of PT, FHA and PRN antibodies in serum of mice were quantitatively determined by the indication degree of TMB color development. The specificity, linear range, accuracy and precision of the developed indirect ELISA detection method were verified. The serum of mice immunized with pertussis vaccine and qualified internal reference products was detected by the developed method.Results The results of the pertussis anti-mouse serum standard and mouse serum samples detected by PT, FHA and PRN antibody ELISA were positive, while the other samples were negative. The linear range of ELISA for PT antibody was 0. 005 3-0. 17 U/mL, the recovery rate was 90. 91%-104. 77%, the RSD of reproducibility verification was 6. 31%, and the RSD among personnel was 6. 40%. The ELISA for FHA antibody had the linear range of 0. 022 3-1. 43 U/mL, the recovery of 95. 84%-102. 18%, the RSD of reproducibility verification of 9. 02%, and the RSD among personnel of 5. 79%. The ELISA for PRN antibody showed the linear range of 0. 009 4-0. 3 U/mL, the recovery rate of 86. 27%-100. 22%, the RSD of reproducibility verification of 6. 94%, and the RSD among personnel of 8. 90%. The detection results of mouse serum immunized with pertussis vaccine and qualified internal reference products by the developed method showed that, the PT antibody levels of six groups of samples were significantly higher than those of the internal reference products, while the FHA and PRN antibody levels were equivalent to those of the internal reference products.Conclusion The developed ELISA methods have strong specificity, good linear correlation(R2> 0. 99), high accuracy and good precision, and can be used for quantitative detection of pertussis(PT, FHA, PRN) antibodies in mouse serum, providing a reference for the optimization of pertussis vaccine production process.

Cardiomyopathies ; Heart Ventricles ; Humans

China ; Humans ; Infant, Newborn ; Intensive Care, Neonatal ; Respiration, Artificial ; Respiratory Care Units

Objective To assess the efficacy and adverse reactions of two cconcentrations of diluted botulinum toxin A for dynamic frontotransverse wrinkles.Methods Totally 103 cases met the inclusion criteria were assigned into two groups by a random number table:high concentration group (50 U/ml,53 cases) and low concentration group (25 U/ml,50 cases).The two groups were given the local injection with 50 U/ml and 25 U/ml concentration of botulinum toxin type A,respectively.Dosage of BTX-A,the treatment effect,the duration of the effect and the incidence of adverse reactions were analyzed.Results The high concentration group were given the BTX-A (45.53±2.05) U,the low concentration group were given the BTX-A (25.71 ± 1.74) U,and the difference were statistically significant (P＜0.05).The differences of post-operative evaluation,time of onset and duration between the two groups were not statistically significant (P＞0.05).In the high concentration group,30 cases (88.24％) had forehead heaviness sensation,and 2 cases (5.71％) in the low concentration group,the difference were statistically significant (P＜0.01).Neither of the groups was given any specific treatment,the symptoms were disappeared in two weeks.In either group there was no serious complications such as intoxication,allergy,etc.Conclusions The efficacy of local injection with two concentrations of diluted botulinum toxin A in the treatment of dynamic frontotransverse wrinkles is positive,the duration of treatment effect between the two groups is similar,and therefore the 25 U/ml concentration is recommended to reduce the forehead heaviness sensation.

Objective To control the hazardous effects of excessive arsenic in drinking water on human health. Methods The experiment on the removal of excessive arsenic from drinking water by electric absorb technology (EST) was performed. Results The pilot experiment and experiment in situ showed that, after treatment by EST, the contents of arsenic in source water(0.06-0.33 mg/L) decreased to the levels of

Cervical intervertebral disc replacement effectively tackles limited motion range of the neck and long-term adjacent degeneration acceleration following traditional anterior cervical fusion and fixation. This paper reviews history of cervical intervertebral disc replacement,current prosthesis types,related indications and complications,and further investigates the feasibility of minimally invasive replacement.

Objective To explore the effects of sports training on endothelin-1 ( ET-1 ) and endothelin-1 receptor (ETR) and its subtypes in the myocardium in rats. Methods After sports training, levels of endothelin-1 were measured using radioimmunoassay, and changes in ETAR and ETBR levels by radioligand binding assay. Results The endothelin-1 level in myocardial cells was significantly reduced ( P

Galectin-9 is a novel eosinophil chemoattractant,which belong to Galectin family.Galectin-9exhibits a variety of biological functions,such as cell differentiation,maturation,adhesion and aggregation,cell chemotaxis,activity,and cell death.Galectin-9 may play important roles in the development of acute inflammation as well as chronic inflammation associated with allergies,autoimmune diseases,infectious processes.This review summarizes the advances in Galectin-9 study.

Objective To construct pEGFP-N1-TGF-β1 recombinant plasmid and transfect it into primary cultured neonatal piglet type Ⅱ alveolar epithelium cell (AEC-Ⅱ) by using lipofectamine 2000,in order to provide basis of methodology for producing recombinant plasmids for transplantation of transfected AEC-Ⅱ into ALI/ARDS animal model lungs.Methods PCR primers were designed to amplify the human TGF-β1 cDNA fragment from plasmid.XhoI and EcoRI were used for double digesting the empty plasmid pEGFP-N1 and cDNA fragment of human TGF-β1.Then the products of double enzyme digestion by using T4 DNA ligase were connected and transformed into DH5α and cultured over night for 16 hours.The structure of recombinant plasmid was identified by using PCR and base sequencing to verify the correctness of pEGFP-N1-TGF-β1 recombinant plasmid.It was then transfected into primarily cultured AEC-Ⅱ by lipofectamine2000 mediated transfection and cultured for another 48 hour.Plasmid DNA (pEGFP-N1-TGF-β1 recombinant plasmid) and lipofectamine 2000 were added into serum-free DMEM respectively,then DNA suspension and Lipofectamine 2000 suspension were blended together and added into cells.After 24-48 hours later,the expression level of enhanced green fluorescent protein (EGFP) was evaluated under fluorescence microscope.Results The structure of vector was verified as pEGFP-N1-TGF-β1 recombinant plasmid by using PCR and base sequencing.Green fluorescence found in some cells showed that the pEGFP-N1-TGF-β1 recombinant plasmids had been successfully transfected into primary cultured AEC-Ⅱ,however,the transfection efficiency still need tobe further improved such as repeating the transfection procedure once again or using adenovirus mediated transfection method to improve the efficiceny and to transplant the cells into animal lungs eventually.Conclusions pEGFP-N1-TGF-β1 recombinant plasmid was successfully constructed and,for the first time,transfected into primarily cultured AEC-Ⅱ of newborn piglets.This established method should be useful for investigation of therapeutic effect and outcomes of lung with experimental acute lung injury.