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Objective To prepare human pneumococcal reference serum and assign IgG antibody concentrations for its 24 serotypes. Methods Fifty healthy male and non-pregnant female volunteers were vaccinated once with 23-valent pneumococcal polysaccharide vaccine. Plasma samples were collected follow-ing immunization and used to prepare reference serum BW09. IgG antibody concentrations of BW09 against the capsular polysaccharides of 24 serotypes (1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, 33F) were assigned by WHO recommended standardized Pn PS ELISA using 09CS (supplied by Lanzhou Institute of Biological Products Co. , Ltd) as a reference se-rum. Finally, 09CS, two quality control sera and eight serum samples had concentrations reassigned for 24 pneumococcal serotypes using BW09 as the reference, and the results were compared with those using 09CS as the reference serum. Results Glycerol-contained human pneumococcal reference serum BW09 was pre-pared and the IgG antibody concentrations for its 24 serotypes were assigned. 09CS had concentrations reas-signed for 24 pneumococcal serotypes using BW09 as the reference, and the newly assigned concentrations were similar to the original values (with percentage error less than 10％ in all serotypes). IgG antibodies against capsular polysaccharides of 24 serotypes of two quality control sera and eight serum samples were as-signed by BW09 and 09CS, and the results showed a linear correlation. Conclusions The human pneumo-coccal reference serum BW09 was successfully prepared and IgG antibody concentrations for its 24 serotypes were accurately assigned.
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Objective: To explore the relationship between NLRP3 inflammasome and atrial fibrillation (AF) by examining peripheral blood level of NLRP3 inlfammasome and other inlfammatory factors in relevant patients. Method: A total of 60 AF patients were enrolled and divided into 2 groups: Paroxysmal AF (PAF) group and Non-paroxysmal atrial fibrillation (nPAF) group, n=30 in each group;in addition, there was a Control group including 26 healthy subjects from physical examination. NLRP3 expression in peripheral blood mononuclear cells (PBMCs) was measured by lfow cytometry;blood levels of IL-1β, IL-6, CRP and NT-proBNP were detected by ELISA. The correlations among different factors were studied by liner regression analysis and the differences were compared among groups. Result:①Compared with Control group, PAF and nPAF groups had increased PBMCs level of NLRP3 and blood levels of IL-1β, IL-6, CRP, NT-proBNP, P0.05.②PAF and nPAF groups showed elevated blood level of NT-proBNP than Control group, P Conclusion: ① NLRP3 inflammasome was closely related to AF, which may provide a therapeutic target for AF treatment. ② AF was closely related to inflammatory response. ③ Downstream product of NLRP3 may cause the inlfammatory response which could induce the occurrence, development and maintenance of AF in relevant patients.
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AIM:To investigate the changes of connexin 43 (Cx43) via establishing a model of sympathomi-metic atrial fibrillation ( AF) .METHODS:The mongrels ( n=15) were randomly divided into control group , rapid atrial pacing (RAP) group and isoprenaline (ISO) perfusion+RAP group (ISO+RAP group).All mongrels’ hearts were taken out rapidly by median sternotomy to establish the cardiac model with Langendorff perfusion in vitro.The atrial effective re-fractory period ( AERP) and AF inducability were tested .The expression and distribution of tyrosine hydroxylase ( TH) were analyzed by immunohistochemistry .Total protein level of Cx 43 and phosphorylation of Cx 43 were determined by West-ern blot.The distribution of Cx43 were also observed by immunofluorescence staining .The cell apoptosis was analyzed by TUNEL staining.The generation of reactive oxygen species ( ROS) in the mitochondria was measured by fluorescence spec-trophotometry .RESULTS:No significant change of AERP was found between control group and RAP group , while that in ISO+RAP group was significantly decreased (P<0.05) and induced AF.Compared with control group, the expression of TH, apoptotic index and the generation of ROS increased gradually (P<0.05), while the content of Cx43 decreased grad-ually both in the total protein and the phosphorylation levels in RAP group and ISO +RAP group (P<0.05).The fluores-cence intensity of Cx43 was also attenuated and Cx43 were lateralized apparently in RAP group , while Cx43 were character-ized as punctate distribution in ISO +RAP group.CONCLUSION:Sympathetic nerves may activate autophagosome at in-tercalated discs and trigger cell apoptosis , resulting in remodeling and downregulation of Cx 43 via oxidative stress , thus having effects on mediating and maintaining AF .
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Objective: To explore the expression of myocardial levels of connexin 43(Cx43), Cx40 in experimental dog model of sympathomimetic atrial fibrillation (AF). Methods: 15 mongrels dogs were randomly divided into 3 groups: Control group, Rapid atrium pacing (RAP) group and RAP+isoprenaline (ISO) perfusion group. n=5 in each group. The hearts were taken to establish in vitro langendorff cardiac perfusion model. Atrial effective refractory period (AERP) and AF inducing rate were tested;intracellular expression and distribution of nerve growth factor (NGF) and tyrosine hydroxylase (TH) were examined by immunohistochemistry, total protein contents of Cx43 and Cx40 were measured by Western blot analysis, mitochondria morphology was observed by transmission electron microscope and mitochondria reactive oxygen species (ROS) generation was detected by fluorescent colorimetric method. Results: AERP was similar between Control group and RAP group (166±5.1) ms vs (160±3.2) ms which cannot induce AF; while it was shortened in RAP+ISO group (148±3.7) ms, P<0.05 which may successfully induce AF.Compared with Control group, mitochondria was slightly swollen in RAP group and the matrix was intact, while mitochondria was obviously swollen in RAP+ISO group and part of matrix was transparent; total protein contents of Cx43 and Cx40 were lower in both RAP group and RAP+ISO group, P<0.05; in addition, they were even lower in RAP+ISO group than RAP group, P<0.05. Compared with Control group and RAP group, RAP+ISO group had increased expression and distribution of NGF, TH and mitochondria ROS generation, P<0.05; NGF, TH and ROS in RAP group were higher than Control group, P<0.05. Conclusion: Sympathetic AF has been related to the contents and changes of myocardial levels of CX43 and Cx40; sympathetic nerve might trigger AF by oxidative stress induced down-regulation of myocardial CX43 and Cx40 in experimental dog model.