A guangxi bama minipig model of trimeresurus stejnegeri snakebite: Modeling and evaluation / 中国组织工程研究

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.

Surgically induced degenerative changes in the femorotibial joints by total medial meniscectomy in minipigs closely resemble late-stage osteoarthritis / 대한수의학회지

Animal models of osteoarthritis (OA) have played a key role in understanding the etiology of OA and in the development of new therapeutic strategies. Although pigs have an advantage as an animal disease model due to their similarity to humans, there are few studies on the induction of OA in minipigs. Therefore, this study aimed to characterize disease progression of OA in total medial meniscectomy (TMM)-operated skeletally mature minipigs, up to day 180 postoperatively. There were no significant alterations in vital signs or hematological indices throughout the observation period. However, clinical manifestations of OA in the medial femoral condyles of TMM-operated minipigs were progressive, depending on postoperative duration, with respect to osteophytes formation and roughened surfaces on radiological observation, cartilage erosion under macroscopic examination, and severe cartilage defects including fibrillation, vertical fissures, and cartilage denuding on histopathological observation, with the highest score indicating late-stage OA on day 180 and without indicating apparent variation between subjects. In particular, the lateral femoral condyles were also degenerated, possibly due to localization of weight-bearing from both menisci to the lateral meniscus. Therefore, TMM in minipigs is suitable for reproducible induction of degenerative changes in the femorotibial joints that closely resemble late-stage OA, and is suitable for use in further research.

Transcriptome Level Changes in Animal Model of Scars During Wound Healing and Pressure Therapy / 医用生物力学

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.

Transcriptome Level Changes in Animal Model of Scars During Wound Healing and Pressure Therapy / 医用生物力学

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.

Magnetic resonance angiography for head and neck arteries of bama minipig / 第二军医大学学报

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.

Establishment of basic data of Bama minipigs for embryo-fetal development toxicity study / 中国比较医学杂志

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.

Mechanism of coagulation dysfunction in the recipients mediated by tissue factor activation after liver xenotransplantation / 器官移植

Objective To investigate the mechanism underlying the activation of tissue factor (TF) that leads to coagulation dysfunction in the recipients after liver xenotransplantation. Methods Auxiliary heterotopic liver xenotransplantation was performed in 3 minipigs with α-1,3-galactosyltransferase gene-knockout (GTKO) as the donors and Tibetan macaque (Macaca thibetana) as the recipients. Postoperative coagulation function changes in the recipients were observed. Reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical staining were adopted to quantitatively measure the expression levels of monkey and minipig TF messenger RNA (mRNA) and protein in the liver tissues of the primary and transplant livers at different time points before and after transplantation. The recalcification time of peripheral blood mononuclear cell (PBMC) was recorded in the normal control monkeys and the recipient monkeys before and 2 h after liver transplantation to evaluate the coagulation status in the recipients. Results All three recipients presented with different degrees of coagulation dysfunction after surgery, manifested as a decrease in fibrinogen level and a reduction in platelet count. The monkey TF protein was positively expressed in the primary livers after surgery, whereas negatively expressed in transplant livers before and after liver transplantation. The minipig TF protein was negatively expressed in both primary livers and transplant livers. At postoperative 2 h, monkey TF mRNA was up-regulated by (2.10±0.24) times in the primary liver compared with the preoperative level, whereas the minipig TF mRNA was up-regulated by (1.42±0.15) times compared with preoperative level. There was statistical significance between the primary livers and transplant livers (P=0.014). Compared with PBMC in the normal control monkeys and recipient monkeys before liver transplantation, the recalcification time of the PBMC in the recipient monkeys was significantly shortened at postoperative 2 h (both P<0.001). Conclusions At the presence of coagulation dysfunction after liver xenotransplantation, the level of TF activation in the primary livers is significantly higher than that in the transplant livers. The TF activation in the primary livers is the main cause of coagulation dysfunction after liver xenotransplantation.

Establishment of a Wuzhishan minipig model of atherosclerosis induced by high fat/cholesterol diet and regulation of Lp-PLA2 expression / 中国实验动物学报

Objective To establish a Wuzhishan minipig model of atherosclerosis(AS) induced by high fat/cholesterol diet,and observe the changes of expression of lipoprotein associated phospholipase A2(Lp-PLA2) in plasma and plaques.Methods 10 Wuzhishan minipigs were randomly divided into 2 groups:The normal control(Ctr,n=4) group was fed with normal diet,and AS model(n=6) group fed with high fat/cholesterol diet for 24 weeks.After the modeling for 24 weeks,the changes of total cholesterol(TC),low density lipoprotein(LDL-C),high density lipoprotein(HDL-C),triglyceride(TG),C-reactive protein(CRP),Lp-PLA2 activity and composition were detected.The changes of vascular lipid deposition and plaques were assessed by pathology using oil red O staining and HE staining,respectively,and immunohistochemical staining for IL-6 protein expression.Moreover,the expression of Lp-PLA2 mRNA determined by RT-PCR and protein by Western blot were observed in the abdominal aortic tissues.Results Compared with the control group,the body weight,body mass index(BMI),TC,LDL-C,HDL-C,CRP,Lp-PLA2 activity and composition and aortic lipid deposition were significantly increased,and AS plaque formation was observed in the AS model group(P<0.05,P<0.01).The expression of Lp-PLA2mRNA and protein and IL-6 protein in abdominal aortic tissues were also significantly increased(P<0.05).Conclusions Long-term high fat/cholesterol diet feeding for 24 weeks can induce atherosclerosis in Wuzhishan minipigs,and Lp-PLA2 plays a key role in the vascular inflammation and plaque formation.

Comparison of organ coefficients and expression of hypoxia-related genes in the Bama and Juema minipigs / 中国比较医学杂志

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.

Influence of high fat diet on insulin resistance and expression of liver insulin receptor substrate 1 and 2 in Tibet minipigs / 中国实验动物学报

Objective To explore the effects of high fat diet on insulin resistance ( IR) and the expression of liver insulin receptor substrate ( IRS) 1 and 2 in Tibet minipigs. Methods Ten Tibet minipigs were randomly divided into 2 groups, normal control (Ctr, n=5) group was fed with normal diet, and IR model (n=5) group fed with high fat/choles-terol diet for 12 weeks. After the establishment of pig models for 12 weeks, the body weight and body length were measured and body mass index ( BMI) was calculated, and the changes of total cholesterol ( TC) , low density lipoprotein ( LDL?C) , high density lipoprotein ( HDL?C) , triglyceride ( TG) , free fatty acids ( FFA) , fasting blood glucose ( FBG) , fasting insu?lin ( insulin) and homeostasis model assessment?insulin resistance ( HOMA?IR) were detected. Glucose tolerance test was performed, the area under the curve of glucose tolerance ( AUC) was also calculated, and the expressions of IRS?1 and IRS?2 gene and protein in liver tissue were detected. The lipid deposition, liver glycogen and pathological changes were ex?amined by pathology using oil?red O, PAS and HE staining, respectively. Results Compared with the control group, the body weight, BMI index, TC, LDL?C, HDL?C, FFA, FBG, insulin and HOMA?IR were significantly increased ( P <0. 05, P<0. 01). Intravenous glucose tolerance test showed that the curve of blood glucose and insulin levels were slowed down, while AUCglucose and AUCinsulin were significantly increased (P<0. 05, P<0. 01). Lipid deposition and liver glyco?gen were increased, and partial hepatocyte swelling, part of the nuclei disappeared or were pushed to one end, occasionally scattered infiltration of lymphocytes in the liver tissue. Furthermore, the expressions of IRS?1 and IRS?2 mRNA and protein were significantly decreased (P<0. 05, P<0. 01). Conclusions High fat diet can induce insulin resistance in Tibet minipigs. The decreased IRS?1 and IRS?2 expression in the liver may be one of the molecular mechanisms involved in the effects of high fat diet on insulin sensitivity in Tibet minipigs.

Observation on the sensitivity of Wuzhishan, Tibetan and Bama minipigs to exogenous fats / 中国比较医学杂志

Objective To observe the sensitivity of Wuzhishan, Tibetan and Bama minipigs to exogenous fats.Methods A total of 15 male minipigs including 5 WZS minipigs, 5 Tibetan minipigs and 5 Bama minipigs, were used in this study.The minipigs were intravenously injected with fat emulsion and fed with high-fat diet, and the changes of serum total cholesterol (TC), low density lipoprotein (LDL-C), high density lipoprotein (HDL-C) and triglyceride (TG) levels were detected at 0.5 h, 1 h, 1.5 h, 2 h, 2.5 h and 3 h after intravenous injection of fat emulsion and at 3 h and 12 h, 24 h, 36 h, 48 h, 60 h, 72 h and 84 h after fed with high-fat diet, respectively, and calculated the changes of area under the curve (ΔAUC) values.Results The triglyceride (TG) in the three kinds minipigs were significantly increased (P Wuzhishan minipig > Bama minipig, while TC, LDL-C and HDL-C showed no significant changes (P>0.05).Moreover, TC, LDL-C, HDL-C and TG were significantly increased in the three stocks of minipigs induced by feeding with high-fat diet (P Bama minipigs > Tibetan minipigs.Conclusions The three stocks of minipigs are sensitive to TG after intravenous injection of fat emulsion, and the lipid tolerance values are in an order of Tibetan minipig > Wuzhishan minipigs > Bama minipigs.Meanwhile, the three stocks of minipigs are also sensitive to TC, LDL-C and HDL-C after feeding with high-fat diet, and the lipid tolerance values are in an increasing order of Wuzhishan minipig > Bama minipigs > Tibetan minipigs.

Cloning, mRNA expression and protein functional bioinformatics analysis of SLA-DOA in Banna mini-pig inbred line / 中国免疫学杂志

Objective:To clone swine leukocyte antigen,class II,DO alpha (SLA-DOA) gene from Banna mini-pig inbred line (BMI) and detect its mRNA expression level in 19 important tissues.Methods:The complete eDNA sequence of SLA-DOA gene was cloned by RT-PCR method from BMI and the mRNA expression pattern in BMI important tissues was examined by semi-quantative RT-PCR method.Nucleotide and protein sequences of SIA-DOA were used to carry out bioinformatics analysis and construct the phylogenetic tree.Results:The eDNA length of BMI SLA-DOA was 1 079 bp,which encoded a protein of 250 amino acids with molecular weight (Mw) 27.81 kD,and isoelectric point (pI) 6.48.Genome structure analysis showed it localized to Sus scrofa chromosomes 7 and consisted of four exons and three introns.Semi-quantitative expression analysis showed that SLA-DOA gene expressed highly in the lymph nodes and stomach;weakly in the heart,skin and duodenum and none in other 14 tissues.Functional bioinformatics analysis indicated that SLA-DOA protein contained one signal poptide,one transmembrane region,three conserved domains,four O-GlcNAc glycosylation sites,18 potential phosphorylation sites and to be located in the cytoplasm with 94.1％ certainty.Phylogenetic analysis demonstrated that BMI (pig) had the closest relationship with cattle.Conclusion:This study have successfully cloned the SLA-DOA gene of Banna mini-pig inbred line,performed bioinformatics analysis and tissue expression profile analysis.It will provide a basis for the studies of BMI xenotransplantation.

An improved method for preparation and evaluation of a porcine model of acute myocardial infarction by minimally invasive surgery and left circumflex artery ligation / 中国实验动物学报

Objective To develop a better method for preparation of porcine model of acute myocardial infarction by permanent occlusion of the left circumflex coronary artery and minimally invasive surgery, evaluate its validity and stability, and explore its application in experimental studies of ischemic heart diseases.Methods 25 healthy female 3-month-old Bama minipigs, body weight 25±3 kg, were used in this study.The porcine model of myocardial infarction was established by minimally invasive surgery and the left circumflex artery ligation at the site of OM1 posterior position under general anesthesia.Heart function was assessed by echocardiography at 15 min before surgery, 1 hour and 4 weeks after surgery.Pathological examination was performed at 4 weeks after the left circumflex artery occlusion.The mortality and cause of death were statistically analyzed.Results The 1-hour and 4-week postoperative cardiac function was considerably decreased, showing a decreased ejection fraction from 64.2±4.6% to 48.2±5.3% (1hour after MI) and 49.7±6.1% (4 weeks after MI) (P<0.01).Pathological examination revealed that the ventricular wall was thinner and the amount of collagens was increased in the infracted area.The ventricular fibrillation rate at 1-hour after myocardial infarction was 17.3% and the infarction area was 19.2%.Conclusions A pig model of acute myocardial infarction can be prepared by our modified left circumflex coronary artery ligation at the obtuse marginal artery (OM1) and minimally invasive surgery.This model exhibits advantages such as minimal surgical trauma, high stability of the model, and low mortality, therefore, provides an ideal and economic animal model for experimental studies on acute ischemic heart diseases.

Establishment of chronic myocardial ischemia model in minipigs and application of noninvasive telemetry technique / 中国比较医学杂志

In order to establish the minipig model of chronic myocardial ischemia and apply non-invasive telemetry technique, the minipig model of chronic myocardial ischemia was made induced by Vitamin D3, isoproterenol and combined with high fat diet, and the non-invasive telemetry technique was used to detect and evaluate the symptoms of minipig model of chronic myocardial ischemia.Moreover, the effects of transport stress and the risk factors of atherosclerosis (AS) induced by high fat diet among Wuzhishan minipigs, Bama minipigs and Tibetan minipigs were also evaluated.Our study has successful established the Bama minipig model of chronic myocardial ischemia and the technical specification for evaluation,.The non-invasive telemetry technique can be used to detect and evaluate the symptoms of chronic myocardial ischemia model, and defines minipigs at least need to keep for more than 4 weeks after transport stress to adaptive recovery period.In addition, the different characteristics of AS risk factors such as hyperlipidemia, hypertension and hyperinsulinemia were observed in Wuzhishan minipigs, Bama minipigs and Tibetan minipigs in high fat environment, and this provides a reference for the selection and application of minipigs in the research of cardiovascular diseases.

Feasibility of transjugular liver biopsy in minipigs / 中国比较医学杂志

Objective To evaluate the feasibility and safety of transjugular liver biopsy( TJLB) by using the LABS 200 liver access and biopsy set ( Cook Inc, USA) .Methods Five minipigs were operated though TJLB puncture under the imaging guidance.The liver biopsies were analyzed by histological examination.Results Technical success of TJLB was achieved in all the 5 minipigs.No procedure-related complications occurred, and sufficient amount of specimen for histological examination was obtained in all cases.Conclusions Our preliminary results indicate that transjugular liver biopsy with the use of Cook LABS 200 liver access and biopsy set is clinically safe and feasible, and provide technical support for its clinical application.

Establishment of a myocardial infarction model and the gene expression profile in peri-infarct area in Juema minipigs / 中国实验动物学报

Objective To establish a Juema minipig model of myocardial infarction, to evaluate the clinical indi?ces in the model pigs, and to explore the relationship between gene expression and metabolic decompensation. Methods 13 male Juema minipigs were randomly divided into control (Sham, n=5), myocardial infarction (MI, n=5) and normal control (for evaluating the recovery condition after surgery, n=3) groups. In the MI group, the ligation was done at the left descending coronary artery around the 1/3 distance to heart apex. Four weeks after the surgery, the cardiac function and serum biochemistry was analyzed. The histological changes and gene expression profiles in the myocardium in the peri?infarct area were exanimated. Results Ultrasonic images showed that the infarction was formed, the ejection fraction and fraction shortening were significantly reduced in the MI group ( ~32% and ~40% less than those of the sham group). Histological examination showed that myocardial fibers at the peri?infarct area were broken, dissolved, and there was con?nective tissue hyperplasia with increased neutrophil and lymphocyte infiltration. Microarray analysis revealed that two myo?cardial remodeling and pathology mediating pathways, three inflammation?related pathways, and 8 metabolic pathways ( in?cluding fatty acid, amino acid, and glucose metabolic pathways) were significantly changed. Conclusions We have suc?cessfully established a Juema minipig model of myocardial infarction. The less branches of the left descending coronary ar?tery allow us to establish a stable model by surgery with comparable characteristics in the clinic indices. The results of this study provides useful reference characteristics of an animal model with characteristic changes in the peri?infarct area.

Comparison of human and animal oral microbiota by Illumina sequencing of 16S rRNA tags / 中国比较医学杂志

Objective To provide original reference data for oral ecosystem research, Tibet minipigs, beagle dogs, rhesus monkey, New Zealand white rabbits and Wistar rats were selected to study their respective characteristics of oral microbial mmunities and compared with normal data of humans.Methods Total DNA was extracted from the specimens of oral microbial communities of Tibet minipigs, beagle dogs, rhesus monkey, New Zealand white rabbits and Wistar rats, and used to amplify 16S rRNA V4 fragments with labeled universal primers.The diversity and structure of microbial communities from those animals were compared with that of humans using BIPES and QIIME analysis after Illumina sequencing of 16S rRNA V4 fragments.Results The richness of the oral microbial communities of humans and the five species of laboratory animals was significantly different (P <0.05).Different species of animals have their own unique oral flora, among which the oral flora of the monkey is the most similar to that of humans.Conclusions Among the five species of laboratory animals, the oral microbial communities of rhesus monkeys and humans have highest similarity. Specifically, the Fusobacterium and Porphyromonas levels of rhesus monkeys is most similar to those of humans.Our findings indicate that rhesus monkeys may be suitable animal model for studies of human oral microbial communities.Tibet minipigs may be suitable animal model for Proteobacteria studies, while beagle dogs may be appropriate for modeling of diseases related to Spirochaetes.

Expression of skeletal muscle glucose and energy metabolism-related genes in Guangxi Bama minipigs with type 2 diabetes mellitus / 中国实验动物学报

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.

Differential regulation of senescence and in vitro differentiation by 17β-estradiol between mesenchymal stem cells derived from male and female mini-pigs

The characterization and potential of mesenchymal stem cells (MSCs) are gender dependent and estrogen influences these properties. This study demonstrated that supplementation with 17β-estradiol (E2) increases the proliferation of bone marrow-MSCs derived from male and female mini-pigs (Mp- and Fp-BMSCs) in a concentration-dependent manner, with 10(-12) M E2 suggested as the optimal dose of E2 that led to the greatest improvement in BMSCs proliferation. Supplementation of 10(-12) M E2 resulted in down-regulation of β-galactosidase activity and pro-apoptotic activity in both BMSCs, while anti-apoptotic activity was up-regulated in only Fp-BMSCs. Further, E2 increased the osteogenic ability of Fp-BMSCs. Based on these findings, optimal utilization of E2 can improve cellular senescence and apoptosis, as well as in vitro osteogenesis of BMSCs, and could therefore be useful in stem cell therapy, particularly in bone regeneration for adult females.

Therapeutic effect of topical application of curcumin during treatment of radiation burns in a mini-pig model

Curcumin protects the skin against radiation-induced epidermal damage and prevents morphological changes induced by irradiation skin, thereby maintaining the epidermal thickness and cell density of basal layers. In this study, the effects of topical curcumin treatment on radiation burns were evaluated in a mini-pig model. Histological and clinical changes were observed five weeks after radiation exposure to the back (⁶⁰Co gamma-radiation, 50 Gy). Curcumin was applied topically to irradiated skin (200 mg/cm²) twice a day for 35 days. Curcumin application decreased the epithelial desquamation after irradiation. Additionally, when compared to the vehicle-treated group, the curcumin-treated group showed reduced expression of cyclooxygenase-2 and nuclear factor-kappaB. Furthermore, irradiation prolonged healing of biopsy wounds in the exposed area, whereas curcumin treatment stimulated wound healing. These results suggest that curcumin can improve epithelial cell survival and recovery in the skin and therefore be used to treat radiation burns.