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Objective To compare the organ coefficients and expressions of hypoxia-related genes in Bama and Juema pigs.Method Real-time quantitative PCR was used to detect the changes of hypoxia gene expressions in the heart,liver,spleen,lung,and kidney of Juema and Bama miniature pigs.Results The organ coefficients of kidney and spleen of Juema pigs were significantly lower than Bama miniature pigs (P<0.05 for both).The heart and lung coefficients of Juema pigs were significantly higher than that of Bama miniature pigs (P<0.05 for both).The VEGF and HIF-1α expressions in the lung and kidney in Juema pigs were significantly higher than Bama pigs (P<0.05 or P<0.01).Only the EPO expression in in the lung of Juema pigs was significantly higher than that of the Bama miniature pigs (P<0.05).Conclusions These results indicate that the variation in organ coefficients may be resulted from evolutionary factors such as adaptiveness to environmental physical and energy conditions,pathogens,and energy metabolism demands,etc.in combination.Juema miniature pigs showing a significantly higher expression of hypoxia-related genes than that in Bama minipigs indicate that it has a strong plateau adaptability by higher gene expressions.
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Objective To investigate the difference of the expression of hypoxia reaction genes of vascular endothelial growth factor(VEGF),erythropoietin (EPO) and hypoxia-inducible factor-1 alpha (HIF-1α)) at different times of rats that were induced epilepsy by kainic acid (KA),and analyze their correlation.Methods The epileptic rat model was established by intraperitoneal injection of kanic acid.The expression of VEGF,EPO and HIF-1α gene were detected by real-time fluorescent quantitative PCR assay with TaqMan probe in different times after intraperitoneal injection of KA.Results Compared with normal sodium (NS) group,the expression VEGF was higher at 12 h((8.38±1.27) ×10-3 ng/μl,P＜0.05)) and 24 h((8.30±5.08) ×10-3 ng/μl,P＜0.05)),EPO was higher at 12 h((8.42±0.90) × 10-5 ng/μl,P＜0.05)) and 48 h ((1.50±3.25) × 10-2 ng/μl,P＜0.01)) while the HIF-1α was higher at 24 h((2.11±0.21) × 10-2 ng/μl,P＜0.01)),48 h((1.50±0.33) × 10-2 ng/μl,P＜0.05))and 72 h((1.64±0.16) × 10-2 ng/μl,P＜0.01)).Furthermore,the expression of EPO showed significant correlations with HIF-1α and VEGF (r=0.573,0.471,P＜0.05),VEGF and HIF-1α had eminent correlations (r=0.803,P＜0.01).Conclusions The expression of VEGF,EPO and HIF-1α participate in the seizure procedure and there is certain correlation between the three genes.
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Objective To compare the Bama minipig and Juema minipig models of high altitude multi-organ dysfunction syndrome.Methods Six plateau-origin Juema minipigs and plain-origin Bama minpigs in each group received intravenous infusion of 0.35 mg/kg lipopolysaccharide ( LPS) , respectively.Blood samples were taken at 0 h, 3 h, 6 h, 12 h, 24 h, 48 h and 72 h after LPS infusion.Routine blood test was performed, blood CK, AST, ALT, TBIL, CRE were assayed, and histopathological examination of the lung tissues was performed at 24 h, 48 h after LPS infusion.Results The mortality of Bama minipigs was 33.3%, higher than that of 16.7%of Juema minipigs.The trend of physiological and biochemical changes was similar, but was milder in the Juema minipigs than in Bama minipigs.The lung injuries of the Bama minipigs at 24 h and 48 h were more severe than those in the Juema minipigs.Conclusions Both Bama and Juema minipig models of high altitude multi-organ dysfunction syndrome can be successfully established.Juema minipig models can be more closely and safely established, due to its own plateau biological properties, and avoid the influence by extrinsic injurious effects of plateau environment.
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Objectives To investigate the correlation between the level of peripheral blood inflammatory cytokines and kainic acid-induced seizure severity in rats.Methods 140 rats were divided into control and model group randomly,70 rats in each group.Model group rats were injected with kainic acid (10 mg/kg) by intraperitoneal,and the control rats were injected with sodium chloride.The change of their behaviors was observed and the concentrations of TNF-α,IL-1β,IL-4 and IL-10 were determined by ELISA in each group at different times.Results The rats showed epilepsy grand mal in 3 h-9 h after KA injection.The concentrations of TNF-α and IL-1β in model groups were significantly higher than those in control group in 6 h-12 h (6 h:(21.5±3.2) pg/ml vs (12.3±3.1)pg/ml;12 h:(20.6±4.2)pg/ml vs (11.5±3.8)pg/ml)(P＜0.05) and IL-4 in model group was higher at only 12 h ((53.55±3.08) pg/ml vs (33.26±4.16)pg/ml) (P＜0.05).The level of IL-10 in model groups was not statistically significant compared with control group (P＞0.05).Conclusion The proinflammatory cytokines (TNF-α and IL-l β) participate in the seizure procedure,meanwhile their levels and seizure severity have eminent correlations,but antiinflammatory cytokines (IL-4 and IL-10) had not.
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Objective To explore the change and its clinical significance of neuron-specific enolase (NSE) and S-100? in serum and hippocampus tissue in rats with epilepsy induced by Kainic acid (KA).Methods 180 Wistar rats were randomly classified into control, KA and carbamazepine (CBZ) group, and the later two groups were further divided into 6 sub-groups (1 h, 4 h, 12 h, 24 h, 48 h and 72 h ) according to epileptic attack at different time point. The NSE and S-100? concentrations in serum and homogenate of hippocampus were determined by radioimmunity assay (RIA) and enzyme linked immunosorbent assay (ELISA), respectively.Results The concentrations of NSE and S-100? were dynamic change during 72 h after epileptic attack in serum and hippocampus homogenate, and the changes were synchronous. The concentrations reached peak at 12 h. Both NSE and S-100? concentrations in KA and CBZ group were obviously higher than those in control group ( P
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Objective To study the specificity, sensitivity of diagnostic antigen for Trichinella spiralis(T. s). Methods T668 recombinant protein from highly efficient expression of newborn larvae stage-specific gene of T. s in E. coli as diagnostic antigen. By using negative and positive sera from rabbit, pig, human as the first antibody, and goat-anti-rabbit IgG, goat-anti-pig IgG, goat-anti-human IgG labeled with HRP as the secondary antibody, indirect ELISA method was established for detecting anti-T.s antibody, while excretory-secretory (ES) antigen of T.s muscle larvae was used as control. Results Sera from rabbit, pig and human were detected by T668 recombinant protein as antigen, which showed a positive rate of 100% with 0.016 ?g/well. There was no difference on the results between the T668 recombinant antigen and the ES antigen for diagnosing T. s-infection. Conclusion The T668 recombinant antigen is promising in substituting ES antigen in the detection of anti-T. s antibody.
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Objective To clone a stage-specific novel cDNA from 5 day-old adult worm (ADS) of Trichinella spiralis. Methods The cDNA library of AD5 was screened by an AD5 stage-specific cDNA probe labeled with digoxigenin (DIG). The positive clones were sequenced and analysed. Results The positive clone contained a cDNA insert of 1 132 bp in length with a full length open reading frame (ORF) of 1 032 bp. The cDNA encoded a polypeptide of 343 amino acid residues(aa) with a molecular weight of 35.1 kDa and an isoelectric point (IP) of 4.8. InterProScan analysis showed that the 117 - 120 aa (SGYG) was a glycosaminoglycan attachment site, 27- 86 aa was nematode cuticle collagen N-terminal domain and 153-228 aa was collagen repeat (G-x-y) domain. Signal PV2.0 analysis indicated that the region of 1-43 aa was a singal peptide. Blastn homology analysis in Genbank revealed that the cDNA had no obvious homology to any other known gene sequence. Blastp analysis revealed high homology to cuticle collagen with identities more than 40 % . Conclusion A novel ADS stage-specific cDNA encoding a full length ORF was cloned and sequence analysis showed this gene encoded cuticle collagen of Trichinella spiralis.