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Objective: To explore the characteristics of Banna miniature pig liver failure induced by amanita exitialis. Methods: From September to October 2020, a reverse high performance liquid chromatography (RP-HPLC) method was used to determine the toxin content of amanita exitialis solution, and 2.0 mg/kg amanita exitialis solution (α-amanitins+β-amanitins) was administered orally to Banna miniature pigs. Toxic symptoms, blood biochemical indexes and histopathological changes of liver, heart and kidney were observed at each time point. Results: All Banna miniature pigs died within 76 h of exposure, and different degrees of digestive tract symptoms such as nausea, vomiting and diarrhea appeared between 6 and 36 h. The biochemical indexes of alanine aminotransferase, aspartate aminotransferase, total bilirubin, lactate dehydrogenase, myoglobin, creatine kinase isoenzyme, blood urea nitrogen and creatinine increased significantly at 52 h after exposure, and the differences were statistically significant compared with 0 h (P<0.05). The bleeding of liver and heart was obvious under macroscopic and microscopic observation, hepatocyte necrosis, renal tubule epithelial cell swelling. Conclusion: Large dose of amanita exitialis can cause acute liver failure of Banna miniature pigs, which is in line with the pathophysiological characteristics of acute liver failure, and lays a foundation for further research on the toxic mechanism and detoxification drugs of amanita exitialis induced liver failure.
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Animals , Swine , Amanitins/metabolism , Swine, Miniature/metabolism , Amanita/metabolism , Liver Failure, Acute , Mushroom Poisoning/diagnosisABSTRACT
Objective To establish a non-venous bypass orthotopic liver transplantation model in Bama miniature pigs with high repeatability and stability. Methods Twelve Bama miniature pigs were randomly divided into the donor group (n=6) and recipient group (n=6). Pigs underwent non-venous bypass orthotopic liver transplantation. The time of anhepatic phase during operation was shortened, blood pressure during anhepatic phase was stably maintained, and management of anesthesia and body fluid during operation were strengthened. The operation time, anhepatic phase and survival status of the recipients were observed and recorded. The intraoperative heart rate, mean arterial pressure (MAP) and changes in arterial blood gas analysis were monitored. The perioperative liver function was evaluated. Results Among 6 Bama miniature pigs, 1 died from transplantation failure intraoperatively. The operation time of the remaining 5 pigs was (247±27) min and the time of anhepatic phase was (46±4) min. Three animals survived for more than 2 weeks. Compared with the preanhepatic phase, the heart rate of the animals was significantly faster, MAP was considerably reduced to (46±6) mmHg, blood pH value, base excess (BE) and HCO3- level were all significantly decreased and serum level of K+ was significantly elevated during the anhepatic phase (all P < 0.05). In the neohepatic phase, MAP of Bama miniature pigs was significantly increased, heart rate was dramatically slower.Blood pH value, BE, HCO3- level were significantly increased and serum level of K+ was significantly declined (all P < 0.05). During abdominal closure, MAP, blood gas indexes and serum level of K+ were almost recovered to those in the preanhepatic phase. Compared with preoperative levels, the levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), lactate dehydrogenase(LDH)and alkaline phosphatase(ALP)were significantly increased after operation (all P < 0.05), the change in AST was the most obvious, and it gradually decreased at postoperative 2 d. The level of γ-gutamyl transferase(GGT) did not significantly elevated. The level of total bilirubin (TB) was evidently elevated at postoperative 5 d. Compared with the preoperative levels, the levels of total protein (TP) and albumin (ALB) were significantly decreased after operation (both P < 0.05), and began to gradually increase at postoperative 1 d. Conclusions The non-venous bypass orthotopic liver transplantation model of Bama miniature pig is convenient, with highly reproducible and survival rate, which can be utilized as a standardized liver transplantation model.
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Objective To observe the changes and significance of the protein expression levels of nuclear factor-κB p65 (NF-κB p65), transforming long factor-β1 (TGF-β1) and apoptosis-related factors Bcl-2 and Bax in myocardial tissue of Bama miniature pig model of diabetic cardiomyopathy (DCM). Methods Ten healthy male Guangxi Bama miniature pigs, aged 4 to 5 months old, were selected and divided into control group and model group according to the random number table method, with 5 pigs in each group. After 12 hours of fasting in the two groups, the DCM model was replicated by intravenous injection of streptozotocin (STZ) 150 mg/kg; for the Bama miniature pigs in the control group, citric acid-sodium citrate buffer 150 mg/kg was injected intravenously. After 10 months of modeling, the basic conditions of the two groups of animals were observed and their fasting blood glucose (FPG) levels were detected. The protein expression levels of NF-κB p65, TGF-β1, Bcl-2 and Bax in myocardial tissue of two groups were detected by Western Blot and the pathological changes of myocardial tissue were observed under electron microscope. Results In the model group, 4 models were successfully established, and 1 died. The model pigs had symptoms such as polydipsia, polyphagia, polyuria and decreased body weight. The FPG level in the model group was significantly higher than that in the control group (mmol/L: 25.53±3.75 vs. 4.68±0.77, P < 0.01). Compared with the control group, the protein expression levels of NF-κB p65, Bax and TGF-β1 in the myocardial tissue of model group were significantly increased (NF-κB p65/GAPDH: 0.46±0.05 vs. 0.38±0.02, Bax/GAPDH: 0.46±0.01 vs. 0.35±0.01, TGF-β1/GAPDH: 0.39±0.01 vs. 0.33±0.01, all P < 0.05) and the expression level of Bcl-2 protein was significantly decreased (Bcl-2/GAPDH: 0.33±0.01 vs. 0.42±0.01, P < 0.01). Electron microscopy results showed that the myofibrils of myocardial tissue in the DCM model group were disordered, and the number of mitochondria in the gap was significantly reduced. A large number of mitochondria with vacuolar degeneration were observed. Conclusions The DCM model of Bama miniature pigs can be successfully replicated after 10 months of high-dose STZ disposable ear vein injection. The DCM model miniature pigs have obvious glucose metabolism disorder, and their myocardial tissue has inflammatory reaction, cardiomyocyte apoptosis and fibrosis.
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Objective To investigate the anatomical structure of arterial vessels in the head and neck of miniature pigs and the related application of vascular cast specimens,and the technology of three-dimensional model reconstruction by CT imaging. Methods A vascular cast specimen of a miniature pig head and neck was made by a 128-slice spiral CT scanning,and a three-dimensional model of the arterial vessels in the head and neck of the miniature pig was reconstructed. Results The cast specimen clearly showed the distribution and running characteristics of the arteries in the head and neck of a miniature pig. The three-dimensional digital model was realistic and stereoscopic,showing the running and distribution of arteries from multiple angles and layers. Conclusions The distribution and running characteristics of arterial vessels in the head and neck of a miniature pig have been investigated by the combination of cast specimen and three-dimensional digital model,providing a morphological reference for the establishment of pig cerebrovascular models in respects of both solid specimen and virtualized model.
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Objective To investigate the optimal condition for the detection of anti-non-galactose (Gal) xenoantigen and antibody in human serum.Mehtods Peripheral blood mononuclear cell (PBMC) obtained from Wuzhishan miniature pig models with α-1,3-galactosyltransferase gene knockout (GTKO) were used as target cells,mixed and incubated with healthy human serum of different concentrations (4.8％,16.7％ and 100％) for 0.5,1.0,2.0,3.0 and 6.0 h,respectively.The abilities of PBMC to bind with IgM and IgG were detected by flow cytometry.Results At the serum concentration of 16.7％,the ability ofnon-Gal IgM to bind with PBMC was significantly enhanced from 0.5 h to 3.0 h incubation (P＜0.01),whereas no statistical significance was noted in terms of IgG (P＞0.05).Increasing serum concentration could also enhance the ability of non-Gal IgM to bind with PBMC.At the serum concentration of 100％ and incubation for 3 h,the ability of IgM to bind with PBMC was the highest among all groups (P＜0.01).At the serum concentration of 100％ and incubation for 6 h,the ability of IgG to bind with PBMC was significantly enhanced (P＜0.05).Prolonging incubation time and increasing serum concentration did not affect the activity of PBMC.Conclusions The optimal condition for detection of anti-non-Gal xenoantigen and antibody is determined.A quantity of 1×105 PBMC from pig should be incubated with 100％ human serum for 3 h for detection of IgM level,or incubated with 100％ human serum for 6 h for measurement of IgG level.This optimized condition contributes to screening the donor pigs which lowly express non-Gal antigen.
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To summarize the breeding and identification of Wuzhishan miniature pig models with α-l,3-galactosyltransferase (GGTA1) gene-knockout (GTKO).Methods The breeding and reproduction perform of GTKO Wuzhishan miniature pigs were assessed and the quantity of piglets was counted.The GTKO Wuzhishan miniature pig models with GGTA1gene knockout were validated by polymerase chain reaction(PCR).The αGal phenotype of peripheral blood mononuclear cells (PBMC) in human,wild-type Wuzhishan miniature pigs and GTKO Wuzhishan miniature pigs was detected by fluorescent microscope and flow cytometry.Routine blood test parameters were statistically compared between the GTKO and wild-type Wuzhishan miniature pigs.Results The inheritance of GGTA1 genotype complied with Mendel's law.Flow cytometry detected no fluorescent expression of PBMC in GGTA1-/-pig models,which were consistent with the genotype identification results.The mean piglets of the primiparous GTKO Wuzhishan miniature pigs were (6.8±1.8) and (8.3±2.2) for the multiparous Wuzhishan miniature pigs.No statistical significance was noted in routine blood test parameters between the GTKO and wild-type Wuzhishan miniature pigs (all P＞0.05).Conclusions Stable inheritance and normal reproductive capacity are observed in two generations of Wuzhishan miniature pigs continuously.GTKO Wuzhishan miniature pig is a reliable donor for heterogeneous organ transplantation.
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To investigate the changes in the expression levels of scavenger receptor class B member 1 (SCARB 1) in the liver tissues before and after liver xenotransplantation and analyze the relationship between the variations in the SCARB1 expression and coagulation regulating dysfunction in the recipients.Methods The Wuzhishan miniature pig with α-1,3-galactosyltransferase gene-knockout(GTKO) was utilized as the donor and Macaca thibetana was chosen as the recipient.Heterotopic auxiliary liver xenotransplantation models were established.The liver tissue specimen was collected before and after liver xenotransplantation.Primary hepatocytes were extracted from the pig using collagenase digestion method.Human peripheral blood mononuclear cells were obtained by immunomagnetic bead sorting.These two types of cells were co-cultured and supplemented with human plasma to establish cell models with coagulation regulating dysfunction following liver xenotransplantation.Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were performed to quantitatively measure and statistically compared the expression levels of messenger ribonucleic acid (mRNA) and protein of SCARB1 in the tissue and cell samples.At the cellular level,the expression of SCARB 1 was interfered by lentiviral vector.The coagulation time was detected to validate the effect upon coagulation function.Results The expression levels of SCARB1 mRNA and protein were significantly down-regulated after liver xenotransplantation (both P＜0.05).In the cell models,the expression levels of SCARB1 mRNA and protein in the porcine hepatocytes co-cultured with human monocytes were significantly down-regulated compared with those in porcine hepatocytes without intervention (both P＜0.05).Compared with the non-intervention group,the coagulation time was significantly prolonged after the expression of SCARB1 was interfered by lentiviral vector (P＜0.05).Conclusions The down-regulated expression of SCARB1 in the liver graft is one of the main causes of mediating coagulation regulating dysfunction.Intervention of SCARB1 expression contributes to resolve the coagulation regulating dysfunction in the recipients after liver xenotransplantation.
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Objective To investigate precise and regulable animal model of benign bile duct strictures.Methods Mini pigs were divided into four groups as sham surgery group, conventional surgery group, small caliber group and large caliber group according to random number table, with 6 pigs in each group.For the small caliber group, outside diameter 3.0mm catheter were inserted into bile duct and partial sutured.For the large caliber group, outside diameter 6.0mm catheter were inserted into bile duct and partial sutured.Conventional surgery group were treated with 5-0 absorbable suture on distal bile duct and controlled stricture severity visually.For sham surgery group, bile duct were exposed without suture.Observe the general condition of four groups.Test liver function on 1st, 3rd, 7th, 14th, 21st and 28th day postoperative.Perform cholangiography on 30th day postoperative.All four groups were put to death to observe the histological changes of liver and bile duct.Results Liver function of sham surgery group showed no obvious change during observation.On 28th day postoperative, compared to conventional surgery group, the liver enzymes ALT, AST, γ-GT, ALP, bilirubin level of small caliber group elevated(P<0.05)and the liver enzymes ALT, AST, γ-GT, bilirubin level of large caliber group elevated(P<0.05),while ALP level lowered(P<0.05).Cholangiography showed bile duct stenosis of different degree on 6 pigs of small caliber group with average diameter 2.7mm (from 2mm to 4mm).As for large caliber group, 5 pigs showed stenosis with average diameter 5.5mm (from 5mm to 7mm).3 pigs of conventional surgery group showed no obvious bile duct stenosis.The 6 pigs of sham surgery group showed no bile duct stenosis.According to designed stenosis standard, the success rate of small caliber group and large caliber group are both higher than conventional surgery group.(P<0.05).30 days after surgery, all 6 pigs of small caliber group had postoperative intraperitoneal adhesion, hepatomegaly, obvious expansion of intrahepatic bile duct and gallbladder expanded by 4~5 times.5 pigs of large caliber group had hepatomegaly, slight dilation of intrahepatic bile duct, obvious expansion of gallbladder, clear choledochal edema, sutured tissue thickening and visible bile duct stenosis.HE staining with the small caliber group showed hepatocyte spotty degenerative necrosis, expanded cholestatic intrahepatic small bile duct, swelling hepatocyte and fibrous septa.HE staining with the large caliber group showed intrahepatic small bile duct slightly expanded and hepatocyte arranged basically normal.VG staining with the small caliber group showed excessive accumulaton of collagen fibers outside bile duct mucous cells, randomly and densely arranged and irregularly shaped.VG staining with the large caliber group showed increased collagen fibers in bile duct submucosa, densely and banded arranged.Conclusion This animal model of mini pig is an relatively ideal large animal model of biliary stricture with high surgical success rate and few postoperative complications.
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In this study, a Specific Pathogen-Free (SPF) Bama miniature pig herd was established.The standards for breeding experimental pigs, genetic test and microbiological quality control had been drafted.The local standards of Heilongjiang province SPF technical specifications for microbiological monitoring of pigs had been formulated.The genetic and microbiological quality criterion had been used to control of the SPF pig herd.Classical swine fever virus (CSFV) and highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) infection models of Bama miniature pigs were established, especially the infection miniature pig model of PRRSV was used to evaluated commercially vaccine.The comparison of the cytokine homology between Bama miniature pigs and domestic pigs showed that the Bama miniature pigs can be used to instead of domestic pigs as an ideal experiental animal.At present the SPF Bama miniature pig colonies have been widely used to study the mechanism of prevention and pathogenic in the classical swine fever virus, porcine reproductive and respiratory syndrome virus and porcine transmissible gastroenteritis virus and porcine circovirus and so on.The completion of the project solved the bottleneck problem of the experimental pig usage which provided a new ideal experimental animal for animal disease and life science research.
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Objective To analyze and evaluate the population genetic quality of 3 subbreeds of China Agricultural University miniature pigs in Beijing.Method According to the local standard DB11/T828.3 -2011, 25 pairs of microsatellite primers were used in 3 subbreeds of China Agricultural University miniature pigs, and software Popgen32 was used to process the data.Results 24 microsatellite loci shared 130, 122 and 138 alleles in the China Agricultural University miniature pigs I, II, III, respectively. The average heterozygosity was 0.6759, 0.5967 and 0.6779, respectively, while the average polymorphism information content ( PIC) was 0.6344, 0.5540 and 0.6403, respectively. The genetic distance between China Agricultural University miniature pig II and III was 0.4251, while the genetic distance between China Agricultural University miniature pig I and II was 0.2084.Conclusions In the 3 subbreeds, China Agricultural University miniature pigs II and III have genetic stability and genetic diversity, and both of which satisfy with the genetic characteristics of closed colony laboratory animal.
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Objective To study the feasibility of Bama miniature pig model establishment of type 2 diabetes mellitus (T2DM) by high-fat and high-sugar diet combined with streptozotocin (STZ), and observe the changes in protein kinase B (PKB) and phosphorylation expressions in skeletal muscle, liver, and pancreatic tissues of the miniature pig model. Methods A total of 10 healthy male Bama miniature pigs were randomly divided into two groups:control group (n=5, fed normal diet) and diabetic model group (n=5, fed high-fat and high-sugar diet combined with STZ to establish T2DM Bama miniature pig model). The fasting plasma glucose (FPG) and fasting insulin (FINS) levels were measured, and insulin resistance (HOMA-IR) was calculated in two groups. The PKB and phosphorylation expressions in skeletal muscle, liver and pancreatic tissues were measured using Western blot assay. Results (1) After 10 months of high-fat and high-sugar diet, the body weight, FPG, FINS and HOMA-IR were significantly higher in model group than those of control group (P<0.01). (2) After STZ treatment, compared with control group, there was a further increased level of FPG and a significantly decreased level of FINS in model group (P<0.01). (3) Compared with control group, the PKB and phosphorylation expression levels in skeletal muscle, liver and pancreas were significantly lower in model group (P<0.05). Conclusion The high-fat and high-sugar diet combined with STZ can successfully establish the T2DM miniature pig model. The PKB and phosphorylation expression levels in skeletal muscle, liver, and pancreatic tissues are decreased in model pigs.
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Objective To study the effect of Sarcandra Glabra on the expression of signal transduction molecules of TGF-β1/Smads signaling pathway in miniature pig of radiation-induced lung injury.Methods 75 miniature pigs were divided into control group,radiation group and radiation plus medication group randomly.At 1 week before exposure of right lung with 15 Gy γ-rays,the miniature pigs in radiation plus medication group were given Sarcandra glabra,while those in the other groups received an equal amount of saline.Right lung were taken at weeks 2,4,8,12 and 24 after irradiation,the pathological changes in the lung tissue were observed by HE staining,and the expression of mRNA and protein of TGF-β1,Smad2,Smad3,and Smad7 were detected by real-time PCR and western blotting,respectively.Results Sarcandra glabra reduced the inflammation and fibrosis of the lung tissue in miniature pig after irradiation.Compared with control group,the expressions of TGF-β1 and Smad3 were significantly increased at 2 weeks after irradiation(P ＜ 0.05),Smad2 and Smad7 were increased at 8 and 12 weeks after irradiation(P ＜ 0.05),respectively,in the radiation group.Compared with the radiation group,the expressions of TGF-β1 and Smad2 were significantly decreased(P ＜ 0.05) from the fourth and eighth week,respectively,Smad3 had no obvious change while Smad7 was significantly increased from the second week in the radiation plus medication group (P ＜ 0.05).Conclusions Sarcandra Glabra plays protective effect on radiation-induced lung injury in miniature pig by regulating TGF-β1,Smad2 and Smad7 expressions in the TGF-β1/Smads signaling pathway.
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Objective To evaluate the novel hybrid liver and renal support system the safety and efficacy of the treatment of Tibet mini pig model of renal failure .Methods Five Tibet miniature pig bilateral renal artery ligation approached to construct the models of renal failure ,treatment group was treated with therapy (CRRT mode) of the novel hybrid liver and renal support system after modeling ,while the control group received no treatment .Drawning venous blood endotoxin ,biochemical tests and aerobic anae‐robic culture when in the treatment of 0 ,4 ,8 h;then we observed the changes of biochemical indicators in Tibet mini pigs before and after treatment ;we recorded machine pressure monitoring indicators every 2 hour during treatment .Results The model of renal failure of two experimental animals were successfully constructed .The general condition of treatment group was improved after treated by the novel hybrid liver and renal support system ,while the control groups continued to deteriorate .The experimental ani‐mals did not appear abnormal ECG during the experiment .Two groups of animals showed no statistics significant difference (P>0 .05) in the period of 0 h .In the remaining period ,heart rate and respiratory rate in treatment group was significantly reduced(P0 .05);after modeling both groups biochemi‐cal indicators was significantly increased compared with baseline (P<0 .05);after treatment of the hybrid liver and renal support systems in the treatment group ,the biochemical parameters were significantly decreased compared with baseline ,the difference was statistically significant different(P<0 .05) ,while the control group did not change significantly .Blood endotoxin measured results were less than 0 .5 EU/mL ,arteriovenous end aerobic anaerobic culture showed no bacterial growth .Pathological examination showed significant swelling of the renal tubular epithelium and tubular necrosis .Conclusion The novel hybrid liver and renal sup‐port system for the treatment of Tibet mini‐pig model of renal failure is safe and effective ,and it can be further attempts for clinical trials .
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Objective To describe the phenotype of NK cells in Bama miniature pigs, and establish an efficient activation and culture method for porcine cytokine-induced killer ( CIK) cells in vitro. Methods The porcine peripheral blood mononuclear cells ( PBMCs) were isolated by Percoll gradient centrifugation, and the phenotype of NK cells was test-ed by detecting the CD2 + /CD8 + /CD3 - cell compartment. To establish an efficient activation and culture method for por-cine CIK cells, we optimized the culture conditions to improve the CIK activation efficiency. Results Using the optimized induction culture conditions, the ratio of CIK ( CD2 + /CD8 + /CD3 -) cells was up to 43. 63% at the fifth day, approxi-mately 5. 59 times increased compared with the initially separated PBMCs. Cell proliferation experiments showed that three obvious fluorescence peaks were observed on the fifth day. The results indicated that the induced CIK cells underwent three times cell division, in theory, about increased 8-fold compared with the initial separation of PBMCs. Furthermore, the qRT-PCR result of the surface markers of porcine NK cells also showed a similar variation tendency as the flow cytometry results. Conclusions Our findings demonstrate the successful establishment of an efficient activation and culture method for porcine CIK cells in vitro.
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Objective To explore the safety of Habib VesOpen bipolar radiofrequency ablation (RFA) catheter used in the treatment of portal vein tumor thrombus (PVTT). Methods A total of 10 miniature pigs were randomly divided into 3 groups. Group A(n=6):RFA of normal portal vein was directly performed;group B (n=2): balloon obstruction of the portal vein was performed first, which was followed by RFA for the fresh thrombus in the portal vein; group C (n=2): PVTT model was established first, and RFA of the portal vein was carried out when the portal thrombus became organized. MRI examination was employed at one, 3 and 4 weeks after RFA; the animals were sacrificed 4 weeks after RFA and pathological examination of portal vein was performed. Results Pigs of group A received portal vein RFA under the condition of 5 W power for 0.6-3.6 min. No obvious abnormality was detected by MRI and pathological examination , which were performed one month after the treatment. In the pigs of group B , MRI performed after RFA showed that the damage of portal vein area was more serious than that in the pigs of group A;abdominal MRI examination performed at one, 3 and 4 weeks after RFA showed that the portal venous edema was gradually decreased;pathological examination at one month after RFA demonstrated serious injury of adjacent liver tissue. Pigs of group C received portal vein RFA under the condition of 7 W power for 1.5 min; no obvious edema of the ablated area was observed on MRI performed after RFA , and pathological examination revealed organized thrombus necrosis and va scular endothelial cell damage. Conclusion When Habib VesOpen bipolar RFA catheter is used for the treatment of PVTT, the RFA power and time should be properly selected according to the severity of PVTT. In order to ensure a safer procedure, high power and short ablation time should be used when the severity of PVTT is mild, while low power and longer ablation time are recommended when the PVTT is more severe.
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The provenances of Rongshui miniature pigs ( RMPs) were purchased from Rongshui, Guangxi, in 2012.130 RMPs were transported to Sanshui, Guangdong,China, which were breed according to the laboratory animal standards.83 RMPs were selected randomly from the first filial generations ( F1 ).The basic data were collected including breed characteristics, reproductive performance, grow curve, hematology, biochemical markers of serum and urine, organ coefficient, Chromosome analysis.According to the national and local standards, the quality control standards of RMP were set up including microbiological, parasitic, environment and facilities, fodder, pathology, genetic.The results showed that RMPs adapt to the climate of Guangdong.The natural mating and conceive rate was 88.3% with the pregnancy of 112 days.The average number of firstborn and still-born was 6.1 and 7.9 respectively.RMP was small body size with the adult body weight of 17.21 ±5.20 kg and 16.35 ±5.23 kg in female and male respectively.RMP was very tame.The mitochondrial genome analysis suggested RMP belonged a typical miniature pig breed in China, which is ancient than Lanyu pig.RMP could be breed as a new kind of laboratory animal.
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Objective Assembly whole mitochondrial genome sequence of Rongshui miniature pig ( RMP ) breed and analysis the structure of mitochondrion based on the next-generation sequecing method.Comparison of phylogenetic relationship and genetic diversity among different pig breeds.Methods We collected peripheral venous blood sample from RMP and constructed two paired-end sequencing libraries.A whole-genome shotgun sequencing strategy and Illumina Genome Analyser sequencing technology were used in our study.Results The mitochondrial genome of RMP consists of 13 protein coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and the length of pig is 16888 bp.The GC content of this pig mitochondrial genome is about 44 %.Based on phlogenetic analysis, population genetic analysis, our findings confirmed that the ancestral cluster in East Asia mainly occurred among Diannan 7#pig, Hainan wild boar, Lanyu and RMP.Conclusion RMP, a typical miniature pig breed in China, is an earlier ancestor than Lanyu pig breed.
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Objective To detect the blood parameter, blood biochemical and electrolyte indices of the F1 generation of Rongshui miniature pig ( RMP) .Methods The blood of 43 female and 42 male RMPs of 4 th month old, and 36 RMPs of 12th month old ( half male and female) were extracted from jugular vein.And the blood parameter, blood biochemical and electrolyte indices were detected by blood analyzer and automatic biochemical analyzer.Results In the same month-old RMP, no significant difference between male and female were found in most indices of blood parameter, blood biochemical and electrolyte indices.On the other hand, many indices were difference between 4th month old and 12th month old RMPs of same gender.Compared with the 4th month old RMP, the 12th month old RMP decreased significantly in WBC and PLT, increased in HGB ( P 0.05 ) .Serum ALT, AST, ALP, CK (male), LDH(male), A/G, BUN, GLU (female), CHOL (male) and K+decreased significantly (P <0.05), while serum TP, TBIL, CR and Ca2+increased significantly (P <0.05),but serum CHOL, TG, HDL-C and LDL-C were not different.86.4%(19/22) biochemical and electrolyte indices in RMP were in/or close to the range of normal value of human.Conclusion Most of the blood parameter, blood biochemical and electrolyte indices of RMP were close to human’ s normal value.
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Objective To establish the data including anatomy and histology of main organs in Rongshui miniature pig (RMP).Methods F1 Rongshui miniature pigs with male and female (2 in each group) in 6 month old were used in this experiment.We measured body weights, dissected these pigs after anaesthesia, recorded total blood volume, total plasma volume, number of spine and dental formula, took main organs for photographs, and made histological sections observed and took photographs by microscope.Results We gained the photographs of main organs and histological sections, organ weights,organic coefficients and other basic data.Conclusion Basic anatomy and histology data of main organs in RMP were collected.
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Organ transplantation is limited by the shortage of human organs. Many studies have sought to overcome this hurdle by using animal organs. Porcine organs, especially from miniature pigs, have been used for organ xenotransplantation rather than nonhuman primates. While the molecular profiling for transplantation is well known in humans and rodents, the situation for pigs is almost completely unknown. The present study examined protein regulation of the developing stages of the pancreatic proteome (4 day-old miniature neonate, 19 day-old miniature piglet, and 14 month-old miniature adult pigs) using two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-time of flight mass spectrometry. Thirteen different expressed spots were observed and nine were identified. The data presented within this study provides critical direction relating to the development of pancreas of miniature pigs, which will assist future proteome analysis of the pancreas, and advance our understanding of the hurdles facing xenotransplantation.