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Objective To study the immunodiagnostic potential of recombinant membrane protein Tetraspanin 2 of Schistosoma japonicum(rSjTsp2) as a diagnostic antigen. Methods The rSjTsp2 was expressed and purified as an antigen used in indirect ELISA to detect specific serum antibody against Schistosoma japonicum of rabbits that at different stages of infection, and the results were compared with those of ELISA using natural antigen (AWA) as antigen. Results The sensitivities of rSjTsp2-ELISA were 75.4% (46/61),77.6% (45/58) and 81.5% (44/54), respectively, 2, 4, 6 weeks after rabbits were infected with Schistosoma japonicum, meanwhile the sensitivities of AWA-ELISA were 68.9% (42/61),86.2% (50/58) and 88.9% (48/54), respectively. The sensitivity difference of two methods was not significant statistically (x2 = 0.652, 1.454,1.174, all P ＞ 0.05).Conclusions The rSjTsp2 is expressed successfully in vitro, and rSjTsp2 recombinant protein expressed in prokaryotic system has a certain application potential in immunodiagnosis.
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Objective To express Schistosoma japonicum Mago nashi(SjMago)gene,and prepare its specific polyclonal antibody.Methods SjMago gene was amplified by PCR from Schistosomulum cDNA library and subcloned into pET28a(+)vector,its recombinant proteins were expressed with IPTG.Rabbits were immunized with the polyacrylamide gel particles containing the recombinant proteins for polyclonal antibody preparation,the sera were detected for antibody specificity by Western blot and titer by ELISA assay.Results SjMago prokaryotic expression plasmid was successfully recombined and the target proteins was induced by IPTG in a molecular weight of 17 X 103，the high titer(1∶40 960)polyclonal antibody was isolated from the immunized rabbit，specific rotein band was detected by Western blot.Conclusion SjMago protein has been successfully expressed and its specific polyantibody is prepared,which lays the foundation for further study.
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<p><b>OBJECTIVE</b>To establish a rat model of heatstroke complicated by endotoxemia for studying the pathogenesis of severe heatstroke.</p><p><b>METHODS</b>Male specific pathogen-free Wistar rats were randomly assigned into 4 groups, namely normothermic saline group (group C), heat exposure saline group (group H), normothermic LPS group (group L), and heat exposure LPS group (group HL). The rectal temperature (Tr), heart rate (HR), mean arterial pressure (MAP), and respiratory rate (RR) of the rats receiving different treatments were continually monitored and their white blood cell count (WBC) and histology of the lungs were observed at 0, 40, 80 and 120 min after the treatments.</p><p><b>RESULTS</b>The rats in HL-Group displayed significantly higher Tr (43.04+/-0.11 degrees C), HR (660+/-42 beats/min), and RR (150+/-11/min) but lower MAP (49.0+/-3.5 mmHg) as compared with the C Group. There were significant differences in the values of Tr, HR, RR and MAP between HL and group L and in HR and MAP between H groups HL and. The rats in group H displayed significantly higher WBC than group C. In contrast, the rats in L groups HL and had significantly lower WBC. LPS injection and heat stress induced pulmonary edema and features characteristic of acute microvascular lung injury in the rats.</p><p><b>CONCLUSION</b>The rat model established by LPS injection and heat stress can successfully mimic the development of severe heatstroke after LPS challenge and heat stress, and provides a suitable model for studying the primordial role of the lungs in the pathogenesis of severe heatstroke.</p>
Subject(s)
Animals , Male , Rats , Disease Models, Animal , Endotoxemia , Heat Stroke , Lipopolysaccharides , Random Allocation , Rats, WistarABSTRACT
<p><b>OBJECTIVE</b>To observe the change in vital signs and arterial blood gas in Lipopolysaccharide (LPS)-injected heat exposed rats.</p><p><b>METHODS</b>Male pathogen-free Wistar rats were randomly assigned to the following groups: saline-injected normothermic control (C-Group), saline-injected heat exposed (H-Group), LPS-injected normothermic control (L-Group), LPS-injected heat exposed (HL-Group). Rectal temperature (Tr), heart rate (HR), mean arterial pressure (MAP), arterial blood gas were continually monitored.</p><p><b>RESULTS</b>(1) The rats in HL-Group displayed significantly high values of Tr (43.04 degrees C +/- 0.11 degrees C) and HR [(660 +/- 42) beats/min] and low values of MAP [(49.0 +/- 3.5) mm Hg] compared with C-Group. There was a significant difference in the values of Tr, HR, and MAP between HL-Group and L-Group and in the values of HR and MAP between HL-Group and H-Group. (2) The values of PaO(2), HCO(3)(-), PaCO(2) were significantly lower than those in C-Group at 40 min after LPS-injected heat stress. At 120 min, the PaO(2) [(11.59 +/- 1.11) kPa], HCO(3)(-) [(10.42 +/- 1.06) mmol/L], PaCO(2) [(2.82 +/- 0.81) kPa] in HL-Group were significantly lower than those in L-Group. A significant difference in the values of HCO(3)(-) and PaCO(2) between HL-Group and H-Group was also observed.</p><p><b>CONCLUSION</b>LPS-injected heat stress primes the rat to advance and augment the change in vital signs, arterial blood gas, and systemic inflammatory response syndrome.</p>