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OBJECTIVE@#To explore the histological structure of the deciduous teeth and the tooth germs of Tibetan miniature pigs for studies of dental tissue diseases and tooth regeneration.@*METHODS@#The structure of the deciduous teeth of Tibetan miniature pigs was observed by X-ray. The ultrastructure of the enamel and dentin of deciduous teeth was characterized by scanning electron microscopy. The jaws and teeth were three-dimensionally reconstructed using Mimics software based on Micro-CT scanning of the deciduous teeth. Image J software was used to calculate the gray value and the mineralization density of the deciduous teeth. Hisotological structure of the tooth germ and the pulp tissue of Tibetan miniature pigs was observed using HE staining.@*RESULTS@#The deciduous teeth of Tibetan miniature pigs were composed of enamel, dentin and medullary pulp tissue. The permanent tooth germ were formed during the deciduous dentition. The enamel and dentin ultrastructure of deciduous teeth were consistent with that of human deciduous teeth. The enamel and dentin mineralization densities were 2.47±0.09 g/cm and 1.72±0.07 g/cm, respectively. The pathological structures of tooth germ and pulp tissue were similar to those of human teeth, and the pulp tissue of the deciduous teeth was in an undifferentiated state.@*CONCLUSIONS@#The deciduous teeth of Tibetan miniature pig have similar anatomy, ultrastructure and histopathological structure to human teeth and can serve as a good animal model for studying human dental tissue diseases and the mechanisms of tooth regeneration.
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Animals , Dental Enamel , Dental Pulp , Dentin , Swine , Swine, Miniature , Tibet , Tooth Germ , Tooth, DeciduousABSTRACT
Objective: To observe the intervening effects of Tongmai Yangxin Pills on miniature pigs with syndrome of deficiency of both qi and yin in later stage of myocardial ischemia reperfusion injury. Methods: A total of 24 miniature pigs were randomly divided into the sham-operated group, the model group (syndrome of deficiency of both qi and yin in myocardial ischemia), the Shengmai Capsules group (0.14 g/kg), and Tongmai Yangxin Pills groups (0.36 g/kg) with six pigs in each group. Except for the sham-operated group, the left anterior descending artery in other groups had been occluded for 30 min by inflation balloons and arterial reperfusion injury was acquires by withdraw balloons. After the operation, the groups were administered with drugs for four weeks. The parameters of noninvasive hemodynamics were measured at the end of study. The tongue color was recorded by digital camera. The tongue and pulse manifestation of the miniature pigs were evaluated according to the symptom-graded scoring method. The size of myocardial ischemia and infarction was analyzed by Evens and TTC staining. Results: Compared with the sham-operated group, the area of myocardial ischemia and necrosis in the model group increased significantly. The noninvasive hemodynamics showed a reduction in cardia coutput (CO) and stroke volume (SV), an augment in systemic vascular resistance (SVR) and left cardiac woke (LCW) of the model group compared with the sham-operated group. Moreover, the scores of tongue and pulse manifestation were also significantly increased in the model group with abnormal color of tongue (P < 0.05, 0.01). Compared with the model group, Shengmai Capsules and Tongmai Yangxin Pills reduced the area of myocardial ischemia and infarction, increased the values of CO and SV, decreased the scores of tongue and pulse manifestation and improved tongue color (P < 0.05, 0.01). In addition, Tongmai Yangxin Pills also reduced SVR (P<0.01). Conclusion: The syndrome of deficiency of both qi and yin in mini pigs could be partly manifested in later stage of myocardial ischemia reperfusion injury. Tongmai Yangxin Pills can improve tongue and pulse signs, protect myocardial tissue, and recover abnormal hemodynamics, which can establish foundation for the further clinical application.
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Objective To explore the feasibility of needle-embedding therapy in the treatment of chronic myocardial ischemia using a miniature pig model established by placement of an Ameroid constrictor at the left anterior descending branch (LAD) of coronary artery in Bama miniature pigs during surgery. Methods The miniature pig model of chronic myocardial ischemia was established by placement of an Ameroid constrictor at the left anterior descending branch of the left coronary artery in Bama miniature pigs. The pig models were randomly divided into the treatment group (the"Neiguan " group) and the control group (the "Zusanli " group), and were treated with needle- embedding electroacupuncture at the"Neiguan" (PC6) and "Zusanli" (ST36) acupoints, respectively. Myocardial samples were taken at 6 weeks after surgery for light and electron microscopic examinations. Results Gross pathology showed that ischemic area in the myocardium appeared in both experimental groups. The ischemic area in the "Zusanli "group was larger than that of the"Neiguan"group. Histopathology showed that the acupuncture treatment at the"Neiguan"acupoint reduced the ischemic injury in the pig myocardial tissues. Ultrastructural observation of the myocardium showed mitochondrial vacuolization in cardiomyocytes and myocardial fibrosis in both groups. Conclusions Acupuncture therapy at the"Neiguan"acupoint of pericardial channel may exert protective effect on the myocardial ischemia by reducing the ischemia-injury of cardiomyocytes, but can not inhibit the already existed ischemia-induced cardiomyocytic injuries. Our findings suggest that the establishment of miniature pig model of chronic myocardial ischemia by surgically placing an Ameroid constrictor on the left anterior descending branch of left coronary artery and the needle-embedding in acupoints is feasible for the treatment of chronic myocardial ischemia in this pig model.
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Objective To explore the effect of general anesthesia on Wuzhishan miniature pigs induced by a mixture of ketamine, Sumianxin II and midazolam, and maintained by ketamine and propofol in surgery lasting up to 8 hours. Methods A total of 18 Wuzhishan miniature pigs (body weight (20. 3 ± 1. 9) kg, 14 male and 4 female) were used in this study. The induction of anesthesia was performed with intramuscular injection of ketamine (8 -10 mg/kg) Sumianxin II (1. 5 mL) and midazolam (10 mg) behind the ear, and the general anesthesia was maintained with a mixture containing 0. 9％ sodium chloride 8 mL, ketamine 100 mg/2 mL and propofol 200 mg/40 mL, continuously injected through the marginal ear vein through a syringe infusion pump. The time spent for anesthesia induction and the duration time of anesthesia were recorded. Physiological indexes including body temperature, blood pressure, heart rate and respiratory rate, the reflex activities, and the effects of analgesia, sedation and muscular relaxation of the miniature pigs under anesthesia at 0, 0. 5, 1, 1. 5, 2, 4, 6, 8 h were observed. Results All the 18 pigs were successfully anaesthetized, but 4 pigs died during surgery due to hypovolemic shock, anesthesia accident, left main coronary thrombosis and reperfusion arrhythmia, respectively. During anesthesia, the analgesia, sedation and muscular relaxation effects on the pigs were obvious. The average time spent for anesthesia induction was (4. 8 ± 1. 2) min and the duration time of anesthesia was (54. 1 ± 5. 8) min. The eyelid reflex, corneal reflex and anal reflex in the pigs were weak or disappeared during 1 -8 h after the anesthesia was induced. The body temperature of the pigs was decreased gradually, with a significant difference between 1 h and 0 h (P＜ 0. 05), reaching the lowest point at 4 h, and then maintained stable. The blood pressure was gradually decreased, reaching the lowest level at 2 h (P ＜ 0. 05), then somehow increased, and maintained at a stable level until the end of surgery. The respiratory rate fluctuated during the anesthesia, with no significant difference. Conclusions The anesthesia induced by a combination of ketamine, Sumianxin II and midazolam and maintained with a combination of ketamine and propofol is simple to operate, shows effects fast, and has good effects of analgesia, sedation and muscular relaxation, keeping the circulatory system and respiratory system relatively stable throughout the anesthesia. Thus it is suitable for general anesthesia for miniature pigs.
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Objective To establish a porcine model of liver failure after different percent hepatectomy.Methods The porcine models of liver failure 75%,85%,95% hepatectomy were developed and the living conditions and survival time were recorded.The blood samples of pre-surgery,post-hepatectomy d1,d3,d5 and post-hepatectomy 1 week,2 weeks,and 3 weeks were collected for hepatic function analysis.Histological examination of liver tissues was performed using HE staining.Liver injury histology was interpreted and scored in the terminal samples.Results The average survival time of pigs with post-hepatectomy liver failure after 75%,85%,95% hepatectomy was 19.0±5.6 days,17.3±5.5 days,1.3±1.5 days,respectively.Their pathological scores were 5.67±0.52,8.17±0.82 and 8.50±0.71,respectively.With the increase of percent hepatic resection,the incidence of hepatic failure was increasing.ALT,AST,ALP,LDH and TBA were dramatically increased in the pigs after 85% hepatectomy.Conclusions The pig model of acute liver failure by 85% hepatectomy is successfully established,which can cause typical acute liver failure in Bama miniature pigs.
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Objective The aim of this study was to explore the effect of androgen deficiency on serum hormone levels, visceral fat accumulation and inflammatory gene expression in miniature pigs fed a high-fat diet ( HFD) . Methods Sexually mature male Chinese Wuzhishan miniature pigs were divided into three groups ( animals/group) as follows:intact male pigs ( SHAM) , castrated male pigs ( CAS) and castrated male pigs plus testosterone treatment ( CAS+T) . The pigs were fed a HFD diet for 12 weeks. Serum levels of testosterone and leptin were measured and visceral fat were dissected and weighted. qRT?PCR was performed to determine the mRNA expression levels of lipogenic, lipolysis and inflammation relat?ed genes. Results (1) Serum testosterone levels were significantly decreased but serum leptin levels were significantly in?creased in the castrated pigs. These effects were recovered after testosterone treatment. ( 2 ) Visceral fat percentage was significantly increased in the castrated pigs, and testosterone treatment reduced the increased visceral fat in the castrated pigs. (3) Castration and testosterone treatment had no significant effects on the expression levels of lipogenic genes (FAS and ACC) and lipolysis genes (HSL and ATGL) in pigs fed a HFD. (5) Castration significantly induced the expressions of inflammatory genes including Leptin, CD68, CCL16, CCL23 and SAA, and testosterone treatment recovered the expres?sions of the above genes in the castrated pigs. Conclusions Castration?induced testosterone deficiency promotes visceral fat accumulation and upregulates the expression levels of inflammatory genes in miniature pigs fed a HFD. Moreover, tes?tosterone treatment ameliorates castration?induced visceral fat accumulation and inflammatory response in HFD?fed pigs.
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Objective To transfect EGFP gene to porcine embryonic fibroblasts ( PEFs) of Tibetan miniature pigs by Lonza Nucleofector II machine and compare the tansfection efficiency between this method and the lipofection method. Method A plasmid carrying green fluorescent protein ( GFP) was transfected into PEFs of Tibetan miniature pigs via the Lonza Nucleofector II machine ( program U020) and by Lipofectamine 2000.Results 5 hours after nucleofection, green fluorescence was observed, indicating 80%transfecting efficiency in the nucleofection group, which is significantly higher than the lipofection group. Conclusion Nucleofector II machine can efficiently transfect PEFs, provides a reliable method for efficiently generate transgenic Tibetan minipigs.
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Objective The aim of this study was to explore the effect of testosterone deficiency on serum lipid levels and hepatic lipid accumulation in miniature pigs fed a high-fat diet (HFD).Methods Eighteen sexually mature male Chinese Wuzhishan miniature pigs (6~7 months old) were used in this study.The pigs were divided in three groups ( n =6 animals/group ) as follows: intact male pigs , castrated male pigs and castrated male pigs with testosterone replacement .They were fed a HFD diet for 12 weeks and body weights were recorded weekly .Serum levels of testosterone , total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and triglycerides (TG) were measured.Hepatic TG and TC levels were also determined , and liver tissues were embedded in paraffin and stained with hematoxylin and eosin (H&E).Results (1) The body weights of pigs in each group were found to be linearly elevated over time .Though castrated pigs gained less weight than did pigs in the other groups , no significant differences were found between them .( 2 ) Castration caused a significant decrease in serum testosterone levels in pigs . This effect was recovered by testosterone treatment .(3) Serum levels of TC, LDL-C and TG were significantly increased in castrated pigs.However, castration had no significant effect on serum HDL-C levels.Testosterone treatment reduced the increased serum lipids in castrated pigs .(4) Hepatic TG and TC contents in castrated pigs were also significantly higher than those in other groups of pigs .Testosterone treatment reduced the increased hepatic lipids in castrated pigs .( 5 ) Compared with other groups of pigs , castrated pigs showed increased steatosis .However , testosterone treatment attenuated hepatic steatosis in castrated pigs .Conclusion Testosterone deficiency caused severe dyslipidemia , and increased hepatic lipid accumulation in miniature pigs fed a high-fat diet.
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Objective To explore the effect and underlying mechanism of Sarcandra glabra Nakai on radiation pneumonnopathy in miniature pigs irradiated with 60Co γ-rays.Methods Totally 60 miniature pigs were randomly divided into blank control group,irradiated group with 15 Gy of 60Co γ-rays,and intervention group administered with Sarcandra glabra Nakai at the dosage of 0.3 g/kg before 15 Gy irradiation.At 4,8,12 and 24 weeks post-irradiation,the respiratory rate and weight were observed,and a portion of the right lung tissue was taken out to conduct HE and Masson staining examination,while the expressions of TGF-β1 and TNF-α as well as the content of hydroxyproline (HP) were detected.Results The respiratory rate,the lung coefficient and the content of HP in the irradiated group showed an increasing trend at 4,8,12 and 24 weeks post-irradiation and significantly higher than those in other two groups (F =21.035,146.014,32.610,P ＜ 0.05).Those indicators were indistinguishable between the control group and intervention group at 4 and 8 weeks post-irradiation (F =0.055,2.456,5.581,P ＞ 0.05),while the indicators of the irradiation group were significantly higher than that of intervention group (F =91.897,93.149,83.487,P ＜0.05) at 12 and 24 weeks post-irradiation.The pulmonary histopathological results showed that no inflammatory response or fibrosis was found in the control group and intervention TGF-β1 and TNF-α in the intervention group were negatively expressed at 4 and 8 weeks post-irradiation and had low expressions at 12 and 24 weeks post-irradiation which was lower than that of the irradiated group.Conclusions Sarcandrae has protective effect against radiation-induced lung injury in miniature pigs probably by inhibiting the expressions of TNF-α and HP.
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Objective To establish small-for-size (SFS) graft injury models in miniature pigs with high standardization, reproducibility and similarity to clinical situation. Methods Ba-Ma miniature pigs were introduced in this study and orthotopic liver transplantations (OLTs) were performed in 12 pigs with 30% liver volume allogeneil grafts (small portion of right paramedian lobe, right lateral lobe and caudate lobe) without veno-venous bypass. The profiles of intra-operational hemodynamics and metabolism were investigated. Animals were observed for 7 days with daily serum biochemistry and coagulation function exam. The survival rate related to operation itself and the SFS grafts were respectively calculated as well as the graft regenerative ratio at post-operational day (POD) 7. Results Graft weight as a percentage of the recipient's native liver weight (GW/RLW) and the total body weight (GW/BW) were (28. 63±4. 42)% and (0. 73±0.06)%. The mean operation time, anhepatic phase, and the time of blockage of infra-hepatic IVC were (191. 7±14. 2) min, (28. 3±3. 6) min, and (45. 0±5. 8) min. The survival rate related to the operation itself and the SFS graft were 83. 33% (10/12) and 40% (4/10), and the graft regenerative ratio at POD7 was (278. 06±42. 95) %. Contrast to the remarkable increase of heart rate and serum potassium during anhepatic phase, the mean arterial pressure, central venous pressure, rectal temperature, PH value and buffer excess had a significant decrease (P<0.01) with a gradual recovery after reperfusioa Serum ALT, AST, PT, Cr, and TB were significantly increased with a peak level at POD1 for the former 4 and POD2 for TB, and then began to decrease and favorably recovered at POD7, but TB, PT, and AST levels were still high when compared to those of prereperfusion (P<0. 05). Conclusion This model of OLT performed with 30% liver volume graft without veno-venous bypass was an ideal large animal model for series studies related to SFS graft injury.
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The miniature pig is a very suitable donor species in xenotransplantation of human organs. Lipid metabolism is an important process that involves the creation and degradation of lipids, which is associated with the function of the gastro-intestinal tract. However, the distribution of lipid metabolism related molecules in the gastro-intestinal tract in the miniature pig is unclear. The present study examined the expression of farnesoid X-receptor (FXR), liver X- receptor (LXR), retinoid X-receptor (RXR), liver fatty acid binding protein (L-FABP), fatty acid synthase (FAS) mRNA in the digestive organs of miniature pigs. FXR and LXR mRNA were not expressed in the stomach but were expressed at high and low density in the small and large intestines, respectively. RXR mRNA was expressed in stomach with moderate density, small intestine with high density and in the large intestine with low density. L-FABP and FAS mRNA were expressed in the stomach and large intestine with low density and in the small intestine with high density. L-FABP mRNA was expressed in the liver and kidney with high density, and in pancreas with low density. FAS mRNA was expressed in the liver with high density, and in pancreas and kidney with low density.
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Humans , Fatty Acid Synthases , Fatty Acid-Binding Proteins , Intestine, Large , Intestine, Small , Intestines , Kidney , Lipid Metabolism , Liver , Pancreas , RNA, Messenger , Stomach , Swine , Tissue Donors , Transplantation, HeterologousABSTRACT
The miniature pig is a very suitable donor species in xenotransplantation of human organs. Lipid metabolism is an important process that involves the creation and degradation of lipids, which is associated with the function of the gastro-intestinal tract. However, the distribution of lipid metabolism related molecules in the gastro-intestinal tract in the miniature pig is unclear. The present study examined the expression of farnesoid X-receptor (FXR), liver X- receptor (LXR), retinoid X-receptor (RXR), liver fatty acid binding protein (L-FABP), fatty acid synthase (FAS) mRNA in the digestive organs of miniature pigs. FXR and LXR mRNA were not expressed in the stomach but were expressed at high and low density in the small and large intestines, respectively. RXR mRNA was expressed in stomach with moderate density, small intestine with high density and in the large intestine with low density. L-FABP and FAS mRNA were expressed in the stomach and large intestine with low density and in the small intestine with high density. L-FABP mRNA was expressed in the liver and kidney with high density, and in pancreas with low density. FAS mRNA was expressed in the liver with high density, and in pancreas and kidney with low density.