ABSTRACT
BACKGROUND: At present, there are not many research methods on the pathological mechanism of Trimeresurus stejnegeri snakebite, and there are few methods for constructing animal models. OBJECTIVE: To establish and evaluate the Guangxi Bama minipig model for research on the pathological mechanism, diagnosis and treatment of Trimeresurus stejnegeri snakebite. METHODS: Based on the 50% lethal dose of intramuscularly injected Trimeresurus stejnegeri venom to mice, the theoretical 50% lethal venom dose for Bama minipigs was calculated by the equivalent dose coefficient conversion and reduction algorithm, and the body surface area conversion algorithm. Twelve Bama minipigs were randomly divided into a normal group (n=6) and a model group (n=6). The model group was injected intramuscularly with 0.2 mL/kg snake venom 1/3 of the theoretical 50% lethal dose (0.643 mg/kg). The control group was injected with the same amount of normal saline. After snake venom injection, the poisoning symptoms of Bama minipigs were observed. Two groups of animal blood samples were collected before, 6 hours and 24 hours after snake venom injection. Blood routine test, four coagulation items, blood biochemistry and electrolyte were detected. Histopathological changes of the heart, brain, lung, liver, and kidney as well as the injection site were observed by hematoxylin-eosin staining. The study protocol was approved by the Laboratory Animal Ethics Committee (approval No. 201909013). RESULTS AND CONCLUSION: There was swelling at the wound of the piglet accompanied with blood blisters after snake venom injection. The affected limbs and the surrounding area were swollen and spread rapidly to the proximal end. The piglets walked all the time because of the pain, and no animal died during the experiment. Compared with the control group pig, the model group had higher red blood cell count, white blood cell count, alanine aminotransferase, D-dimer count, longer prothrombin time, and lower fibrinogen and platelet count. Hematoxylin-eosin staining showed that, compared with the control group, capillary permeability of piglet lung tissues in the model group was increased with hyperemia and edema. Edema, bleeding, degeneration and necrosis were seen in the muscle tissue on the injection site. No obvious abnormalities in other organs and tissues were observed. To conclude, this method can be used to establish a pig model of Trimeresurus stejnegeri snakebite that can reflect the pathophysiological process of Trimeresurus stejnegeri snakebite. It has operability and repeatability that can be used to study the pathophysiological mechanism of Trimeresurus stejnegeri snakebite.
ABSTRACT
Objective To study the changes of transcriptome levels in a Bama minipig model of hypertrophic scar during wound healing and pressure therapy by using RNA sequencing (RNA-seq) technique. Methods The Bama minipig model was established by skin wounds from the back and pressure (3.4 kPa) was initiated at 60 days after skin injury. Total RNA was extracted from scar tissues at 0, 14, 30, 60, and 90 days after skin injury and then sequenced. The resulting sequences were mapped to porcine reference genomes and transcriptomes were reconstructed to search for differentially expressed genes (DEGs). The DEGs were further subjected to GO and KEGG analysis using bioinformatics method, while part of the genes were selected for verification using qRT-PCR. Results After preprocessing, more than 78% reads in each group were accurately aligned to the reference sequence. The DEGs identification result showed that 568 genes were differentially expressed after pressure treatment, with 289 up-regulated and 279 down-regulated. GO enrichment analysis revealed that DEGs in each group were mainly associated with extracellular matrix, tissue development and skin development. KEGG analysis showed that the DEGs in each group during wound healing were mainly enriched in extracellular matrix-receptor interactions, focal adhesion and apoptosis pathways; while the DEGs after pressure treatment were mainly enriched in PI3K-Akt and MAPK signaling pathway except the pathways mentioned. qRT-PCR showed that the expression patterns of 6 DEGs were consistent with RNA-seq analysis, confirming the reliability of RNA-seq result. Conclusions RNA-Seq analysis identified differentially expressed genes in animal model of scars during wound healing and pressure therapy, which provided experimental evidence for clinical scar treatment.
ABSTRACT
Objective To study the changes of transcriptome levels in a Bama minipig model of hypertrophic scar during wound healing and pressure therapy by using RNA sequencing (RNA-seq) technique. Methods The Bama minipig model was established by skin wounds from the back and pressure (3.4 kPa) was initiated at 60 days after skin injury. Total RNA was extracted from scar tissues at 0, 14, 30, 60, and 90 days after skin injury and then sequenced. The resulting sequences were mapped to porcine reference genomes and transcriptomes were reconstructed to search for differentially expressed genes (DEGs). The DEGs were further subjected to GO and KEGG analysis using bioinformatics method, while part of the genes were selected for verification using qRT-PCR. Results After preprocessing, more than 78% reads in each group were accurately aligned to the reference sequence. The DEGs identification result showed that 568 genes were differentially expressed after pressure treatment, with 289 up-regulated and 279 down-regulated. GO enrichment analysis revealed that DEGs in each group were mainly associated with extracellular matrix, tissue development and skin development. KEGG analysis showed that the DEGs in each group during wound healing were mainly enriched in extracellular matrix-receptor interactions, focal adhesion and apoptosis pathways; while the DEGs after pressure treatment were mainly enriched in PI3K-Akt and MAPK signaling pathway except the pathways mentioned. qRT-PCR showed that the expression patterns of 6 DEGs were consistent with RNA-seq analysis, confirming the reliability of RNA-seq result. Conclusions RNA-Seq analysis identified differentially expressed genes in animal model of scars during wound healing and pressure therapy, which provided experimental evidence for clinical scar treatment.
ABSTRACT
Objective To display the normal vascular structures of head and neck of Bama minipig by three-dimensional time-of-flight (3D TOF) and time-resolved imaging of contrast kinetics (TRICKS) magnetic resonance angiography (MRA), and to compare and analyze the imaging indicators of arteries, so as to provide references for large animal models of cerebrovascular disease. Methods 3D TOF and TRICKS MRA of head and neck vessels in 3 healthy Bama minipigs were performed using a 3.0 T magnetic resonance system. The artery definition scores of the two MRA methods were compared, and the contrast ratio (CR), signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of the main arteries were calculated for quantitative analysis. Results In terms of artery definition, 3D TOF MRA and TRICKS MRA both achieved high scores for the common carotid arteries and external carotid arterial system, but low scores for posterior circulation. 3D TOF MRA was significantly better than TRICKS MRA in displaying the intracranial artery system (1.60±0.50 vs 1.37±0.49, P＝0.019 8). Quantitative analysis showed that TRICKS MRA improved CR of arteries to different extents; however, there were no significant differences in SNR or CNR between 3D TOF and TRICKS MRA (P0.05). Conclusion Both 3D TOF and TRICKS MRA have good and comparable diagnostic performance for common carotid arteries and their main branch structures of Bama minipig, and each method has its advantages and limitations.
ABSTRACT
Objective To obtain the basic data of Bama minipigs and provide basic reference for embryo-fetal developmental toxicity study. Methods Pregnant minipigs were sacrificed at different days during the gestation period respectively. The examinations included necropsy,count of corporea lutea,live and dead implantations,fetal body weight, and external,visceral, and skeletal examination of fetuses. Results The basic data of Bama minipigs, such as body weight, fetal development, and fetal malformation/variation were obtained. Conclusions We obtained the basic reproductive parameters of pregnant Bama minipigs and the indexes of fetal development, which can provide valuable reference data for embryo-fetal developmental toxicity tests of Bama minipigs.
ABSTRACT
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.
ABSTRACT
Objective In this study, the glucose and energy metabolism-related genes (PGC-1α, Glut-4, ERRα, NRF-1, TFAM and mtDNA gene) were detected in type 2 diabetes mellitus ( T2DM) and non-T2DM minipigs, and the gene function was explored for T2DM pathogenesis.Methods The longissimus muscle of T2DM and non-T2DM Guangxi Bama mini-pigs was used as experiment material.The expression of glucose and energy metabolism-related genes was detec-ted by QRT-PCR.Results The expressions of PGC-1α, Glut-4, ERRαand NRF-1 genes were significantly higher than that of non-T2DM group, the expressions of TFAM and mtDNA gene were lower than that of non-T2DM group.Conclu-sions The upregulated expression of PGC-1αgene and its downstream genes Glut-4, ERRα, NRF-1 may improve the glu-cose metabolic functions in skeletal muscle in the Bama minipigs, whereas insufficient mitochondrial synthesis may induce decreasing ATP synthesis, and results in skeletal muscle insulin resistance, finally leading to the T2DM occurrence.
ABSTRACT
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.
ABSTRACT
Objective To clone the coding sequence of Guangxi Bama mini-pig PGC-1αgene, and to analyze the expression of PGC-1αgene in various tissues of mini-pigs using RT-PCR and QRT-PCR techniques.Methods The PGC-1αgene coding sequence ( CDS) was amplified by PCR from the cDNA of longissimus muscle of Guangxi Bama mini-pig. The PCR products were inserted into pEASY-T5 vector, transfected E.coli, identified and sequenced.The PGC-1αgene expression in different tissues of the Bama mini-pigs was detected by RT-PCR and QRT-PCR assays.Results The PGC-1αgene CDS of Guangxi Bama mini-pig was cloned.It was 2391 bp in length.It had 99.9%homology with the reference sequence, and had two synonymous mutations that were C-A1105 and G-A1524.The expression level of PGC-1αgene was higher in the heart and kidney, followed by liver, subcutaneous fat and longissimus muscle, but the expression was not de-tected in pancreas of Guangxi Bama mini-pig.Conclusions We have successfully cloned the PGC-1αgene of Guangxi Bama mini-pig, and detected this gene expression in six tissues.The results of this study will provide a basis for studying the effect of PGC-1αon type 2 diabetes mellitus (T2DM) in Bama mini-pigs.
ABSTRACT
Objective To measure the circadian rhythm of consecutive monitoring electrocardiogram ( ECG ) , blood pressure, respiration, activities in Bama minipigs by telemetry technology .Methods Six 6-month-old Bama minipigs were implanted with VAP port in Superficial femoral artery , after recovery 7 days , 24 hours ECG , blood pressure, activity and respiratory parameters were recorded and analyzed in the EMAK noninvasive telemetry system . Results The ECG, blood pressure, respiration and activities in Bama minipigs showed circadian rhythm changes , Bama minipigs daytime heart rate was significantly higher than the nighttime heart rate (P <0.01), and daytime PR interval , QRS interval and QT interval was significantly lower than the nighttime (P <0.05, P <0.01), daytime mean heart rate was 76.22 beats/min, nighttime mean heart rate was 67.03 night beats/min;daytime mean PR interval , QRS interval andQT interval were 109.97 ms, 42.72 ms and 380.37 ms, nighttime mean PR interval , QRS interval and QT interval were 112.32ms, 44.01ms and 389.24ms.Bama minipigs daytime systolic blood pressure , diastolic blood pressure and mean blood pressure were significantly higher than the nighttime ( P <0.01 ) , daytime mean systolic blood pressure , diastolic blood pressure and mean blood pressure were 129.57 mmHg, 96.75 mmHg and 111.73 mmHg, nighttime mean systolic blood pressure , diastolic blood pressure , mean blood pressure were 122.81 mmHg, 92.65 mmHg and 106.19 mmHg, and the nighttime reduction rate of systolic blood pressure , diastolic blood pressure and mean blood pressure were 19.89%, 19.05%and 19.35%.In addition , Bama minipigs daytime activity and respiratory rate were significantly higher than the nighttime (P <0.01).Conclusion It’s Feasible to measure the circadian rhythm of electrocardiogram (ECG), blood pressure, activity and respiratory parameters in Bama minipigs by telemetry technology , and can truly shows those physiological parameters of the Bama minipigs within 24h, to provide a reference for pharmacology and toxicology research by using Bama minipigs .