Traumatic brain injury(TBI) at a young age can lead to the development of long-term functional impairments. Severity of injury is well demonstrated to have a strong influence on the extent of functional impairments;ho...Traumatic brain injury(TBI) at a young age can lead to the development of long-term functional impairments. Severity of injury is well demonstrated to have a strong influence on the extent of functional impairments;however, identification of specific magnetic resonance imaging(MRI) biomarkers that are most reflective of injury severity and functional prognosis remain elusive. Therefore, the objective of this study was to utilize advanced statistical approaches to identify clinically relevant MRI biomarkers and predict functional outcomes using MRI metrics in a translational large animal piglet TBI model. TBI was induced via controlled cortical impact and multiparametric MRI was performed at 24 hours and 12 weeks post-TBI using T1-weighted, T2-weighted, T2-weighted fluid attenuated inversion recovery, diffusion-weighted imaging, and diffusion tensor imaging. Changes in spatiotemporal gait parameters were also assessed using an automated gait mat at 24 hours and 12 weeks post-TBI. Principal component analysis was performed to determine the MRI metrics and spatiotemporal gait parameters that explain the largest sources of variation within the datasets. We found that linear combinations of lesion size and midline shift acquired using T2-weighted imaging explained most of the variability of the data at both 24 hours and 12 weeks post-TBI. In addition, linear combinations of velocity, cadence, and stride length were found to explain most of the gait data variability at 24 hours and 12 weeks post-TBI. Linear regression analysis was performed to determine if MRI metrics are predictive of changes in gait. We found that both lesion size and midline shift are significantly correlated with decreases in stride and step length. These results from this study provide an important first step at identifying relevant MRI and functional biomarkers that are predictive of functional outcomes in a clinically relevant piglet TBI model. This study was approved by the University of Georgia Institutional Animal Care and Use Committee(AUP: A2015 11-001) on December 22, 2015.展开更多
AIM: To establish a new pig model for auxiliary partial orthotopic liver transplantation (APOLT).METHODS: The liver of the donor was removed from its body. The left lobe of the liver was resected in vivo and the right...AIM: To establish a new pig model for auxiliary partial orthotopic liver transplantation (APOLT).METHODS: The liver of the donor was removed from its body. The left lobe of the liver was resected in vivo and the right lobe was used as a graft. After the left lateral lobe of the recipient was resected, end-to-side anastomoses of suprahepatic inferior vena cava and portal vein were performed between the donor and recipient livers,respectively. End-to-end anastomoses were made between hepatic artery of graft and splenic artery of the host.Outside drainage was placed in donor common bile duct.RESULTS: Models of APOLT were established in 5 pigs with a success rate of 80%. Color ultrasound examination showed an increase of blood flow of graft on 5th d compared to the first day after operation. When animals were killed on the 5th d after operation, thrombosis of hepatic vein (HV) and portal vein (PV) were not found. Histopathological examination of liver samples revealed evidence of damage with mild steatosis and sporadic necrotic hepatocytes and focal hepatic lobules structure disorganized in graft. Infiltration of inflammatory cells was mild in portal or central vein area. Hematologic laboratory values and blood chemical findings revealed that compared with group A (before transplantation), mean arterial pressure (MAP), central venous pressure (CVP), buffer base (BB), standard bicarbonate (SB) and K+ in group B (after portal vein was clamped) decreased (P＜0.01). After reperfusion of the graft, MAP, CVP and K+ restored gradually.CONCLUSION: Significant decrease of congestion in portal vein and shortened blocking time were obtained because of the application of in vitro veno-venous bypass during complete vascular clamping. This new procedure,with such advantages as simple vessel processing, quality anastomosis, less postoperative hemorrhage and higher success rate, effectively prevents ischemia reperfusion injury of the host liver and deserves to be spread.展开更多
AIM： To evaluate the effect of double-balloon enteros- copy （DBE） on pancreas histology and levels of pancre- atic enzymes. METHODS： Conventional upper gastrointestinal endos- copy was performed on five control pi...AIM： To evaluate the effect of double-balloon enteros- copy （DBE） on pancreas histology and levels of pancre- atic enzymes. METHODS： Conventional upper gastrointestinal endos- copy was performed on five control pigs. Oral DBE was performed with an EN-450T5 enteroscope on 20 pigs. Two experimental groups （10 pigs each） were defined according to DBE duration： 90 rain for Group 1 and 140 min for Group 2. During oral insertion, the balloons were not inflated in the descending part of the duodenum to avoid the minor duodenal papilla. Serum amy- lase, lipase and C-reactive protein （CRP） levels were monitored before the procedure and repeated every 30 min until the exploration was finished, as well as 24 h and 7 d after. After the procedure and for a total of 7 d, the pigs were observed twice a day for signs of de- creased activity, irritability, vomiting or anorexia. Gross and microscopic examination of the pancreas was per- formed on day 7. RESULTS： All animals tolerated DBE without clini- cal manifestations of acute pancreatitis. Experimental groups had higher levels of enzymes than the control group at 24 h. Throughout the exploration, the amylase levels increased significantly above the baseline 24 h after DBE, although the increase was not statistically significant and did not reach 20% of the baseline. An increase in lipase and CRP was observed at 24 h after the procedure, although by day 7, all enzymatic lev- els had returned to baseline. No differences between Groups 1 and 2 were found for any enzyme and sam- pling site during and after the procedure. Similarly, no correlation between insertion depth and enzyme levels was observed. Direct in situ and post-removal inspec- tion of the pancreas did not show any evidence of fluid collection, abscesses or hemorrhage. Histological examination of the pancreas from Groups 1 and 2 re- vealed the existence of focal areas （0.14-0.26 mm2） of ischemic necrosis in 47.4% of the animals. In the pigs with damaged pancreas, the left lobe （tail） was always affected. However, this only happened in 83.3% of the samples from the right lobe （head） and in 33.3% of the samples from the body of the pancreas. Significant differences were found between the left lobe （tail） and the body for the percentage of affected pancreas. Both the size of the lesions and the percentage of affected pancreas were higher in the left pancreatic lobe （tail）. The presence of the lesions was not related to the ex- ploration length.CONCLUSION： The increase in pancreatic enzymes after DBE could be related to focal points of pancreatic ischemic necrosis due to mechanical stress.展开更多
Mini pig models are large mammals and their ears are more similar with human beings in structure and development than other animals.However,the study on porcine ears is still in the initial stage and there is no descr...Mini pig models are large mammals and their ears are more similar with human beings in structure and development than other animals.However,the study on porcine ears is still in the initial stage and there is no description of an ideal operation approach to endocochlear potential and potassium ion concentration measurements.In this article,we describe a pre-auricular surgical approach to access the middle and inner ear for endocochlear potential and potassium ion concentration measures in mini pig models.Ten one-week old normal mini pigs were used in the study.The bulla of the temporal bone was accessed via a pre-auricular approach for endocochlear potential and potassium ion concentration measurements.The condition of the animals during the first post—experiment 24 h was observed.One animal died during surgery.The preauricular approach improved protection and preservation of relevant nervous and vascular elements including the facial nerve and carotid artery.So,the pre-auricular approach can be used for endocochlear potential and potassium ion concentration measurements with improved nerve and artery preservation mini pigs.展开更多
Background:Male infertility is an important issue that causes low production in the animal industry.To solve the male fertility crisis in the animal industry,the prediction of sperm quality is the most important step....Background:Male infertility is an important issue that causes low production in the animal industry.To solve the male fertility crisis in the animal industry,the prediction of sperm quality is the most important step.Sperm RNA is the potential marker for male fertility prediction.We hypothesized that the expression of functional genes related to fertilization will be the best target for male fertility prediction markers.To investigate optimum male fertility prediction marker,we compared target genes expression level and a wide range of field data acquired from artificial insemination of boar semen.Results:Among the genes related to acrosomal vesicle exocytosis and sperm–oocyte fusion,equatorin(EQTN),zona pellucida sperm-binding protein 4(ZP4),and sperm acrosome membrane-associated protein 3 exhibited high accuracy(70%,90%,and 70%,respectively)as markers to evaluate male fertility.Combinations of EQTN-ZP4,ZP4-protein unc-13 homolog B,and ZP4-regulating synaptic membrane exocytosis protein 1(RIMS1)showed the highest prediction value,and all these markers are involved in the acrosome reaction.Conclusion:The EQTN-ZP4 model was efficient in clustering the high-fertility group and may be useful for selection of animal that has superior fertility in the livestock industry.Compared to the EQTN-ZP4 model,the ZP4-RIMS1 model was more efficient in clustering the low-fertility group and may be useful in the diagnosis of male infertility in humans and other animals.The appointed translational animal model and established biomarker combination can be widely used in various scientific fields such as biomedical science.展开更多
Magnetic resonance imaging(MRI)is a clinically relevant,real-time imaging modality that is frequently utilized to assess stroke type and severity.However,specific MRI biomarkers that can be used to predict long-term f...Magnetic resonance imaging(MRI)is a clinically relevant,real-time imaging modality that is frequently utilized to assess stroke type and severity.However,specific MRI biomarkers that can be used to predict long-term functional recovery are still a critical need.Consequently,the present study sought to examine the prognostic value of commonly utilized MRI parameters to predict functional outcomes in a porcine model of ischemic stroke.Stroke was induced via permanent middle cerebral artery occlusion.At 24 hours post-stroke,MRI analysis revealed focal ischemic lesions,decreased diffusivity,hemispheric swelling,and white matter degradation.Functional deficits including behavioral abnormalities in open field and novel object exploration as well as spatiotemporal gait impairments were observed at 4 weeks post-stroke.Gaussian graphical models identified specific MRI outputs and functional recovery variables,including white matter integrity and gait performance,that exhibited strong conditional dependencies.Canonical correlation analysis revealed a prognostic relationship between lesion volume and white matter integrity and novel object exploration and gait performance.Consequently,these analyses may also have the potential of predicting patient recovery at chronic time points as pigs and humans share many anatomical similarities(e.g.,white matter composition)that have proven to be critical in ischemic stroke pathophysiology.The study was approved by the University of Georgia(UGA)Institutional Animal Care and Use Committee(IACUC;Protocol Number:A2014-07-021-Y3-A11 and 2018-01-029-Y1-A5)on November 22,2017.展开更多
Meniscus injuries are widespread and the available treatments do not offer enough healing potential.Here,we provide critical support for using pigs as a biological model for meniscal degeneration and the development o...Meniscus injuries are widespread and the available treatments do not offer enough healing potential.Here,we provide critical support for using pigs as a biological model for meniscal degeneration and the development of cutting-edge therapies in orthopedics.We present a single-cell transcriptome atlas of the meniscus,consisting of cell clusters corresponding to four major cell types:chondrocytes,endothelial cells,smooth muscle cells,and immune cells.Five distinct chondrocyte subclusters(CH0–CH4)were annotated,of which only one was widespread in both the red and white zones,indicating a major difference in the cellular makeup of the zones.Subclusters distinct to the white zone appear responsible for cartilage-specific matrix deposition and protection against adverse microenvironmental factors,while those in the red zone exhibit characteristics of mesenchymal stem cells and are more likely to proliferate and migrate.Additionally,they induce remodeling actions in other chondrocyte subclusters and promote the proliferation and maturation of endothelial cells,inducing healing and vascularization processes.Considering that they have substantial remodeling capabilities,these subclusters should be of great interest for tissue engineering studies.We also show that the cellular makeup of the pig meniscus is comparable to that of humans,which supports the use of pigs as a model in orthopedic therapy development.展开更多
BACKGROUND Colonic anastomotic leakage(AL)remains a feared complication of colorectal surgery.Usually,a defunctioning stoma or a proximal colostomy is performed to reduce the AL rate but cannot completely prevent AL.M...BACKGROUND Colonic anastomotic leakage(AL)remains a feared complication of colorectal surgery.Usually,a defunctioning stoma or a proximal colostomy is performed to reduce the AL rate but cannot completely prevent AL.Moreover,defunctioning colostomy is associated with high morbidity.This study assessed the feasibility of completely preventing colonic AL using total enteric flow diversion without a defunctioning stoma in a pig model of colonic AL.AIM To determine the feasibility of preventing colonic AL via total enteric flow diversion in pigs.METHODS A total of 14 pigs underwent surgery to create colon anaesthesia with severe defects for establishing the AL model.The pigs were then randomized into the control group(n=7),which received no further therapy,and a diversion group(n=7),which underwent placement of a modified ileostomy tube to divert the enteric contents from the colon externally.The general condition,serum Creactive protein level,white blood cell count,5-day incidence of colon AL,and development of abdominal abscesses were evaluated.RESULTS A modified ileostomy tube with a balloon was placed and pressurized to 20 kPa at a distance of 10-20 cm proximal to the ileocecal valve,effectively obstructing the intestine without causing injury and efficiently diverting the enteric contents.In the diversion group,no cases of peritonitis or abscess were observed.In contrast,all pigs in the control group developed either abdominal abscesses or peritonitis.CONCLUSION Instead of ileostomy or colostomy,the total enteric flow diversion technique with the placement of a modified ileostomy tube and balloon in the ileum can effectively or completely prevent colon AL.展开更多
Background:Previous studies have shown that exogenous short-chain fatty acids(SCFAs)introduction attenuated the body fat deposition in conventional mice and pigs.However,limited studies have evaluated the effects of e...Background:Previous studies have shown that exogenous short-chain fatty acids(SCFAs)introduction attenuated the body fat deposition in conventional mice and pigs.However,limited studies have evaluated the effects of exogenously introduced SCFAs on the lipid and glucose metabolism independently of the gut microbiota.This study was to investigate the effects of exogenous introduction of SCFAs on the lipid and glucose metabolism in a germ-free(GF)pig model.Methods:Twelve hysterectomy-derived newborn pigs were reared in six sterile isolators.All pigs were hand-fed with sterile milk powder for 21 d,then the sterile feed was introduced to pigs for another 21 d.In the second 21-d period,six pigs were orally administrated with 25 mL/kg sterile saline per day and considered as the GF group,while the other six pigs were orally administrated with 25 mL/kg SCFAs mixture(acetic,propionic,and butyric acids,45,15,and 11 mmol/L,respectively)per day and regarded as FA group.Results:Orally administrated with SCFAs tended to increase the adiponectin concentration in serum,enhance the CPT-1 activity in longissimus dorsi,and upregulate the ANGPTL4 mRNA expression level in colon(P<0.10).Meanwhile,the mRNA abundances of ACC,FAS,and SREBP-1C in liver and CD36 in longissimus dorsi of the FA group were decreased(P<0.05)compared with those in the GF group.Besides,the mRNA expression of PGC-1αin liver and LPL in longissimus dorsi tended to(P<0.10)upregulate and downregulate respectively in the FA group.Moreover,oral administration of SCFAs tended to increase the protein level of GPR43(P<0.10)and decrease the protein level of ACC(P<0.10)in liver.Also,oral administration of SCFAs upregulated the p-AMPK/AMPK ratio and the mRNA expressions of GLUT-2 and GYS2 in liver(P<0.05).In addition,the metabolic pathway associated with the biosynthesis of unsaturated fatty acids was most significantly promoted(P<0.05)by oral administration of SCFAs.Conclusions:Exogenous introduction of SCFAs might attenuate the fat deposition and to some extent improve the glucose control in the pig model,which occurred independently of the gut microbiota.展开更多
Background Cold regions have long autumn and winter seasons and low ambient temperatures.When pigs are unable to adjust to the cold,oxidative damage and inflammation may develop.However,the differences between cold an...Background Cold regions have long autumn and winter seasons and low ambient temperatures.When pigs are unable to adjust to the cold,oxidative damage and inflammation may develop.However,the differences between cold and non-cold adaptation regarding glucose and lipid metabolism,gut microbiota and colonic mucosal immunological features in pigs are unknown.This study revealed the glucose and lipid metabolic responses and the dual role of gut microbiota in pigs during cold and non-cold adaptation.Moreover,the regulatory effects of dietary glucose supplements on glucose and lipid metabolism and the colonic mucosal barrier were evaluated in cold-exposed pigs.Results Cold and non-cold-adapted models were established by Min and Yorkshire pigs.Our results exhibited that cold exposure induced glucose overconsumption in non-cold-adapted pig models(Yorkshire pigs),decreasing plasma glucose concentrations.In this case,cold exposure enhanced the ATGL and CPT-1αexpression to promote liver lipolysis and fatty acid oxidation.Meanwhile,the two probiotics(Collinsella and Bifidobacterium)depletion and the enrichment of two pathogens(Sutterella and Escherichia-Shigella)in colonic microbiota are not conducive to colonic mucosal immunity.However,glucagon-mediated hepatic glycogenolysis in cold-adapted pig models(Min pigs)maintained the stability of glucose homeostasis during cold exposure.It contributed to the gut microbiota(including the enrichment of the Rikenellaceae RC9 gut group,[Eubacterium]coprostanoligenes group and WCHB1-41)that favored cold-adapted metabolism.Conclusions The results of both models indicate that the gut microbiota during cold adaptation contributes to the protection of the colonic mucosa.During non-cold adaptation,cold-induced glucose overconsumption promotes thermogenesis through lipolysis,but interferes with the gut microbiome and colonic mucosal immunity.Furthermore,glucagon-mediated hepatic glycogenolysis contributes to glucose homeostasis during cold exposure.展开更多
Objective: To investigate the electrophysiological changes of autonomic cells in left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with chronic heart failure.Methods: Guinea pigs mo...Objective: To investigate the electrophysiological changes of autonomic cells in left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with chronic heart failure.Methods: Guinea pigs model of iron deficiency anemia complicated with chronic heart failure in 10 guinea pigs of the experimental group was made by feeding a low iron diet,pure water and subcutaneous injection of isoproterenol. The control group consisting of 11 guinea pigs was given normal food, normal water and injected with normal saline. The left ventricular outflow tract model specimen was also prepared. The standard microelectrode technique was used to observe electrophysiological changes of autonomic cells in the outflow tract of left ventricular heart failure complicated with iron deficiency anemia in guinea pig model. The indicators of observation were maximal diastolic potential, action potential amplitude, 0 phase maximal depolarization velocity, 4 phase automatic depolarization velocity, repolarization 50% and 90%, and spontaneous discharge frequency.Results: Compared with the control group, 4 phase automatic depolarization velocity,spontaneous discharge frequency and 0 phase maximal depolarization velocity decreased significantly(P < 0.01) and action potential amplitude reduced(P < 0.01) in model group. Moreover, repolarization 50% and 90% increased(P < 0.01).Conclusions: There are electrophysiological abnormalities of the left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with heart failure.展开更多
Functional magnetic resonance imaging(fMRI) is em-ployed in many behavior analysis studies, with blood oxygen level dependent-(BOLD-) contrast imaging being the main method used to generate images. The use of BOLD-con...Functional magnetic resonance imaging(fMRI) is em-ployed in many behavior analysis studies, with blood oxygen level dependent-(BOLD-) contrast imaging being the main method used to generate images. The use of BOLD-contrast imaging in f MRI has been refined over the years, for example, the inclusion of a spin echo pulse and increased magnetic strength were shown to produce better recorded images. Taking careful precautions to control variables during measurement, comparisons between different specimen groups can be illustrated by f MRI imaging using both quantitative and qualitative methods. Differences have been observed in comparisons of active and resting, developing and aging, and defective and damaged brains in various studies. However, cognitive studies using f MRI still face a number of challenges in interpretation that can only be overcome by imaging large numbers of samples. Furthermore, f MRI studies of brain cancer, lesions and other brain pathologies of both humans and animals are still to be explored.展开更多
AIM： To investigate the effects of green flickering light on refractive development and expression of muscarinic acetylcholine receptor（mAChR） M1 in the eyes of guinea pigs.METHODS： Thirty guinea pigs（15-20 days ...AIM： To investigate the effects of green flickering light on refractive development and expression of muscarinic acetylcholine receptor（mAChR） M1 in the eyes of guinea pigs.METHODS： Thirty guinea pigs（15-20 days old） were randomly divided into three groups（n=10/group）. Animals in group I were raised in a completely closed carton with green flickering light illumination. Those in group II were kept in the open top closed carton under normal natural light. Guinea pigs were raised in a sight-widen cage under normal natural light in group III. The refractive status and axial length were measured before and after 8 weeks＇ illumination. Moreover, total RNA extracted from retinal, choroidal, and scleral tissues were determined by real-time reverse transcription polymerase chain reaction（RT-PCR）. The expressions of the receptor M1 were also explored in the retina, choroid, and sclera using immunohistochemistry.RESULTS： There was a remarkable reduction in refractive error and increase in axial length after 8-weeks＇ green flickering light stimulation（P〈0.001）. The expression of M1 receptor mRNA in sclera and retina in myopia group were remarkably lower than that in group II and III（P〈0.01）. Significant reduced expression of M1 receptor stimulated by green flickering light in retina and sclera tissues were also observed（P〈0.05）. However, there was no M1 receptor expression in choroid in 3 groups.CONCLUSION： Myopia can be induced by 8 weeks＇ green flickering light exposure in the animal model. M1 receptor may be involved causally or protectively in myopia development.展开更多
To apply a new airway treatment to humans, preclinical studies in an appropriate animalmodel is needed. Canine, porcine and leporine tracheas have been employed as animal airwaystenosis models using various methods su...To apply a new airway treatment to humans, preclinical studies in an appropriate animalmodel is needed. Canine, porcine and leporine tracheas have been employed as animal airwaystenosis models using various methods such as chemical caustic agents, laser, and electrocautery.However, existing models take a long time to develop (3- 8 weeks) and the mechanism of stenosisis different from that in humans. The aim of the present study was to establish a new and fasttracheal stenosis model in pigs using a combination of cuff overpressure intubation (COI) andelectrocautery. Fourteen pigs were divided into three groups: tracheal cautery (TC) group (n=3),COI group (n=3), and COI-TC combination group (n=8). Cuff overpressure (200/400/500 mmHg)was applied using a 9-mm endotracheal tube. Tracheal cautery (40/60 watts) was performed usinga rigid bronchoscopic electrocoagulator. After intervention, the pigs were observed for 3 weeks andbronchoscopy was performed every 7 days. When the cross-sectional area decreased by > 50%, itwas confirmed that tracheal stenosis was established. The time for tracheal stenosis was 14 days inthe TC group and 7 days in the COI-TC combination group. In the COI group, no stenosis occurred.In the COI-TC group, electrocautery (40 watts) immediately after intubation for>1 h with a cufpressure of 200 mmHg or more resulted in suficient tracheal stenosis within 7 days. Moreover, thedegree of tracheal stenosis increased in proportion to the cuff pressure and tracheal intubation time.The combined use of cuf overpressure and electrocautery helped to establish tracheal stenosis inpigs rapidly.展开更多
This study was conducted to investigate host-microbiota interactions and explore the effects of maternal gut microbiota transplantation on the growth and intestinal functions of newborns in a germ-free(GF)pig model.Tw...This study was conducted to investigate host-microbiota interactions and explore the effects of maternal gut microbiota transplantation on the growth and intestinal functions of newborns in a germ-free(GF)pig model.Twelve hysterectomy-derived GF Bama piglets were reared in 6 sterile isolators.Among them,6 were considered as the GF group,and the other 6 were orally inoculated with healthy sow fecal suspension as fecal microbiota transplanted(FMT)group.Another 6 piglets from natural birth were regarded as the conventional(CV)group.The GF and FMT groups were hand-fed with Co60-y-irradiated sterile milk powder,while the CV group was reared by lactating Bama sows.All groups were fed for 21 days.Then,all piglets and then were switched to sterile feed for another 21 days.Results showed that the growth performance,nutrient digestibility,and concentrations of short-chain fatty acids in the GF group decreased(P<0.05).Meanwhile,the serum urea nitrogen concentration and digesta pH values in the GF group increased compared with those in the FMT and CV groups(P<0.05).Compared with the CV group,the GF group demonstrated upregulation in the mRNA expression levels of intestinal barrier function-related genes in the small intestine(P<0.05).In addition,the mRNA abundances of intestinal development and absorption-related genes in the small intestine and colon were higher in the GF group than in the CV and FMT groups(P<0.05).The FMT group exhibited greater growth performance,lipase activity,and nutrient digestibility(P<0.05),higher mRNA expression levels of intestinal development and barrier-related genes in the small intestine(P<0.05),and lower mRNA abundances of pro-inflammatory factor in the colon and jejunum(P<0.05)than the CV group.In conclusion,the absence of gut microbes impaired the growth and nutrient digestibility,and healthy sow gut microbiota transplantation increased the growth and nutrient digestibility and improved the intestinal development and barrier function of newborn piglets,indicating the importance of intestinal microbes for intestinal development and functions.展开更多
Inducible expression systems are indispensable for precise regulation and in-depth analysis of biological process.Binary Tet-On system has been widely employed to regulate transgenic expression by doxycycline.Previous...Inducible expression systems are indispensable for precise regulation and in-depth analysis of biological process.Binary Tet-On system has been widely employed to regulate transgenic expression by doxycycline.Previous pig models with tetracycline regulatory elements were generated through random integration.This process often resulted in uncertain expression and unpredictable phenotypes,thus hindering their applications.Here,by precise knock-in of binary Tet-On 3G elements into Rosa26 and Hipp11 locus,respectively,a double knock-in reporter pig model was generated.We characterized excellent properties of this system for controllable transgenic expression both in vitro and in vivo.Two att P sites were arranged to flank the td Tomato to switch reporter gene.Single or multiple gene replacement was efficiently and faithfully achieved in fetal fibroblasts and nuclear transfer embryos.To display the flexible application of this system,we generated a pig strain with Dox-inducing h KRASexpression through phiC31 integrase-mediated cassette exchange.After eight months of Dox administration,squamous cell carcinoma developed in the nose,mouth,and scrotum,which indicated this pig strain could serve as an ideal large animal model to study tumorigenesis.Overall,the established pig models with controllable and switchable transgene expression system will provide a facilitating platform for transgenic and biomedical research.展开更多
Harlequin ichthyosis (HI) is a severe genetic skin disorder and caused by mutation in the ATP-binding cassette A12 (ABCA12) gene. The retinoid administration has dramatically improved long-term survival of HI, but imp...Harlequin ichthyosis (HI) is a severe genetic skin disorder and caused by mutation in the ATP-binding cassette A12 (ABCA12) gene. The retinoid administration has dramatically improved long-term survival of HI, but improvements are still needed. However, the ABCA12 null mice failed to respond to retinoid treatment, which impedes the development of novel cure strategies for HI. Here we generated an ethylnitrosourea mutagenic HI pig model (named Z9), which carries a novel deep intronic mutation IVS49-727 A>G in the ABCA12 gene, resulting in abnormal mRNA splicing and truncated protein production. Z9 pigs exhibit significant clinical symptom as human patients with HI. Most importantly, systemic retinoid treatment significantly prolonged the life span of the mutant pigs via improving epidermal maturation, decreasing epidermal apoptosis, and triggering the expression of ABCA6. Taken together, this pig model perfectly resembles the clinical symptom and molecular pathology of patients with HI and will be useful for understanding mechanistic insight and developing therapeutic strategies.展开更多
In a recent study,Yong-Guang Yang and colleagues developed a severely immunodeficient pig model that supports long-term engraftment and multilineage differentiation of human hematopoietic stem/progenitor cells(HSPCs)....In a recent study,Yong-Guang Yang and colleagues developed a severely immunodeficient pig model that supports long-term engraftment and multilineage differentiation of human hematopoietic stem/progenitor cells(HSPCs).1 This breakthrough addresses key limitations of current humanized mouse models,including their small size and limited lifespan,and offers a promising platform for the large-scale production of human immune cells,potentially functioning as an in vivo bioreactor.展开更多
Metabolites and metabolism-related gene expression profiles in skeletal muscle change dramatically under obesity,aging and metabolic disease.Since obese and lean pigs are ideal models for metabolic research.Here,we co...Metabolites and metabolism-related gene expression profiles in skeletal muscle change dramatically under obesity,aging and metabolic disease.Since obese and lean pigs are ideal models for metabolic research.Here,we compared metabolome and transcriptome of Longissimus dorsi(LD)muscle between Taoyuan black(TB,obese)and Duroc(lean)pigs at different ages.We defined the“window phase”of intramuscular fat(IMF)deposition in TB pig,which has significantly higher IMF than Duroc pig.Our results displayed discrepant lipid composition and different expression genes(DEGs)enriched in lipid metabolism,and both metabolome and transcriptome analyses revealed stronger energy expenditure and more active amino acid and protein metabolism in Duroc pig.10 up-and 51 down-regulated biomarker metabolites with age-and breed-specificity were identified.Potential secretory metabolites,including organic acid(fumaric acid,succinate,malic acid,and gamma-aminobutyric acid),amino acid(L-lysine,and L-glutamic acid),lipid(2-hydroxyisovaleric acid,and L-carnitine)were demonstrated a significant correlation with IMF deposition.Our research highlights the huge difference of metabolic spectrum in skeletal muscle between obese and lean model and muscle-derived secretory metabolites might act as an ambassador of intercellular communication to regulate systematic metabolism.展开更多
Background and purpose The low-field MRI is a promising tool to accurately diagnose strokes.We here report our study on the accuracy of a 0.23-Tesla(0.23-T)MRI using the haematoma enhanced inversion recovery(HEIR)sequ...Background and purpose The low-field MRI is a promising tool to accurately diagnose strokes.We here report our study on the accuracy of a 0.23-Tesla(0.23-T)MRI using the haematoma enhanced inversion recovery(HEIR)sequence to detect acute ischaemic stroke(AIS)and intracerebral haemorrhage(ICH)within 24 hours of symptom onset.Methods A novel HEIR sequence based on fluid-attenuated inversion recovery T1-weighted,with a scanning time of 1 min and 17 s,was developed using an ICH and AIS pig model on a 0.23-T MRI.Images of the pig model were obtained hourly for 24 hours in order to monitor value changes on T1/T2 and verify the differential diagnosis of AIS and ICH.Then,30 patients with AIS and 30 patients with ICH with confirmed diagnoses by 3T-MRI/CT were included.Diagnostic criteria on a 0.23-T MRI for ICH was the hyperintensity signal on both the diffusion-weighted imaging(DWI)and HEIR sequence,while for AIS was the hyperintensity on DWI and isointensity on the HEIR sequence.Two blinded raters independently assessed the images obtained by the 0.23-T MRI for the presence of ICH/AIS.Results In the pig model,setting the inversion time to 800 ms enabled clear differentiation of ICH from brain parenchymal tissue and AIS.In real patients,a correct 0.23-T MRI diagnosis of either an AIS or ICH was made in all 60 patients within 24 hours of symptom onset(100%overall accuracy).No adverse events occurred.Conclusions The 0.23-T MRI may have the potential to differentiate cerebral haemorrhage from cerebral infarction with both speed and accuracy,making brain MRI scans easier,faster and cheaper.It might be possible to improve the screening imaging process for strokes in the emergency room.Further multicentre studies are needed to validate our findings.展开更多
基金Financial support was provided by the University of Georgia Office of the Vice President for Research to FDW。
文摘Traumatic brain injury(TBI) at a young age can lead to the development of long-term functional impairments. Severity of injury is well demonstrated to have a strong influence on the extent of functional impairments;however, identification of specific magnetic resonance imaging(MRI) biomarkers that are most reflective of injury severity and functional prognosis remain elusive. Therefore, the objective of this study was to utilize advanced statistical approaches to identify clinically relevant MRI biomarkers and predict functional outcomes using MRI metrics in a translational large animal piglet TBI model. TBI was induced via controlled cortical impact and multiparametric MRI was performed at 24 hours and 12 weeks post-TBI using T1-weighted, T2-weighted, T2-weighted fluid attenuated inversion recovery, diffusion-weighted imaging, and diffusion tensor imaging. Changes in spatiotemporal gait parameters were also assessed using an automated gait mat at 24 hours and 12 weeks post-TBI. Principal component analysis was performed to determine the MRI metrics and spatiotemporal gait parameters that explain the largest sources of variation within the datasets. We found that linear combinations of lesion size and midline shift acquired using T2-weighted imaging explained most of the variability of the data at both 24 hours and 12 weeks post-TBI. In addition, linear combinations of velocity, cadence, and stride length were found to explain most of the gait data variability at 24 hours and 12 weeks post-TBI. Linear regression analysis was performed to determine if MRI metrics are predictive of changes in gait. We found that both lesion size and midline shift are significantly correlated with decreases in stride and step length. These results from this study provide an important first step at identifying relevant MRI and functional biomarkers that are predictive of functional outcomes in a clinically relevant piglet TBI model. This study was approved by the University of Georgia Institutional Animal Care and Use Committee(AUP: A2015 11-001) on December 22, 2015.
文摘AIM: To establish a new pig model for auxiliary partial orthotopic liver transplantation (APOLT).METHODS: The liver of the donor was removed from its body. The left lobe of the liver was resected in vivo and the right lobe was used as a graft. After the left lateral lobe of the recipient was resected, end-to-side anastomoses of suprahepatic inferior vena cava and portal vein were performed between the donor and recipient livers,respectively. End-to-end anastomoses were made between hepatic artery of graft and splenic artery of the host.Outside drainage was placed in donor common bile duct.RESULTS: Models of APOLT were established in 5 pigs with a success rate of 80%. Color ultrasound examination showed an increase of blood flow of graft on 5th d compared to the first day after operation. When animals were killed on the 5th d after operation, thrombosis of hepatic vein (HV) and portal vein (PV) were not found. Histopathological examination of liver samples revealed evidence of damage with mild steatosis and sporadic necrotic hepatocytes and focal hepatic lobules structure disorganized in graft. Infiltration of inflammatory cells was mild in portal or central vein area. Hematologic laboratory values and blood chemical findings revealed that compared with group A (before transplantation), mean arterial pressure (MAP), central venous pressure (CVP), buffer base (BB), standard bicarbonate (SB) and K+ in group B (after portal vein was clamped) decreased (P＜0.01). After reperfusion of the graft, MAP, CVP and K+ restored gradually.CONCLUSION: Significant decrease of congestion in portal vein and shortened blocking time were obtained because of the application of in vitro veno-venous bypass during complete vascular clamping. This new procedure,with such advantages as simple vessel processing, quality anastomosis, less postoperative hemorrhage and higher success rate, effectively prevents ischemia reperfusion injury of the host liver and deserves to be spread.
基金Supported by PI070712 Project (Instituto de Salud Carlos Ⅲ, Ministerio de Sandiady Consumo, Spain)BIO-MED07/08-0019 Project (Consejería de Educación, Cienciae Investigación de la Región de Murcia, Spain)
文摘AIM： To evaluate the effect of double-balloon enteros- copy （DBE） on pancreas histology and levels of pancre- atic enzymes. METHODS： Conventional upper gastrointestinal endos- copy was performed on five control pigs. Oral DBE was performed with an EN-450T5 enteroscope on 20 pigs. Two experimental groups （10 pigs each） were defined according to DBE duration： 90 rain for Group 1 and 140 min for Group 2. During oral insertion, the balloons were not inflated in the descending part of the duodenum to avoid the minor duodenal papilla. Serum amy- lase, lipase and C-reactive protein （CRP） levels were monitored before the procedure and repeated every 30 min until the exploration was finished, as well as 24 h and 7 d after. After the procedure and for a total of 7 d, the pigs were observed twice a day for signs of de- creased activity, irritability, vomiting or anorexia. Gross and microscopic examination of the pancreas was per- formed on day 7. RESULTS： All animals tolerated DBE without clini- cal manifestations of acute pancreatitis. Experimental groups had higher levels of enzymes than the control group at 24 h. Throughout the exploration, the amylase levels increased significantly above the baseline 24 h after DBE, although the increase was not statistically significant and did not reach 20% of the baseline. An increase in lipase and CRP was observed at 24 h after the procedure, although by day 7, all enzymatic lev- els had returned to baseline. No differences between Groups 1 and 2 were found for any enzyme and sam- pling site during and after the procedure. Similarly, no correlation between insertion depth and enzyme levels was observed. Direct in situ and post-removal inspec- tion of the pancreas did not show any evidence of fluid collection, abscesses or hemorrhage. Histological examination of the pancreas from Groups 1 and 2 re- vealed the existence of focal areas （0.14-0.26 mm2） of ischemic necrosis in 47.4% of the animals. In the pigs with damaged pancreas, the left lobe （tail） was always affected. However, this only happened in 83.3% of the samples from the right lobe （head） and in 33.3% of the samples from the body of the pancreas. Significant differences were found between the left lobe （tail） and the body for the percentage of affected pancreas. Both the size of the lesions and the percentage of affected pancreas were higher in the left pancreatic lobe （tail）. The presence of the lesions was not related to the ex- ploration length.CONCLUSION： The increase in pancreatic enzymes after DBE could be related to focal points of pancreatic ischemic necrosis due to mechanical stress.
基金supported by grants from the National Basic Research Program of China(973 Program)(2012CB967900. 2012CB967901)the National Nature Science Foundation of China(NSFC81400472)
文摘Mini pig models are large mammals and their ears are more similar with human beings in structure and development than other animals.However,the study on porcine ears is still in the initial stage and there is no description of an ideal operation approach to endocochlear potential and potassium ion concentration measurements.In this article,we describe a pre-auricular surgical approach to access the middle and inner ear for endocochlear potential and potassium ion concentration measures in mini pig models.Ten one-week old normal mini pigs were used in the study.The bulla of the temporal bone was accessed via a pre-auricular approach for endocochlear potential and potassium ion concentration measurements.The condition of the animals during the first post—experiment 24 h was observed.One animal died during surgery.The preauricular approach improved protection and preservation of relevant nervous and vascular elements including the facial nerve and carotid artery.So,the pre-auricular approach can be used for endocochlear potential and potassium ion concentration measurements with improved nerve and artery preservation mini pigs.
基金supported by the Basic Science Research Program through the National Research Foundation of Korea(NRF),funded by the Ministry of Education(NRF-2018R1A6A1A03025159).
文摘Background:Male infertility is an important issue that causes low production in the animal industry.To solve the male fertility crisis in the animal industry,the prediction of sperm quality is the most important step.Sperm RNA is the potential marker for male fertility prediction.We hypothesized that the expression of functional genes related to fertilization will be the best target for male fertility prediction markers.To investigate optimum male fertility prediction marker,we compared target genes expression level and a wide range of field data acquired from artificial insemination of boar semen.Results:Among the genes related to acrosomal vesicle exocytosis and sperm–oocyte fusion,equatorin(EQTN),zona pellucida sperm-binding protein 4(ZP4),and sperm acrosome membrane-associated protein 3 exhibited high accuracy(70%,90%,and 70%,respectively)as markers to evaluate male fertility.Combinations of EQTN-ZP4,ZP4-protein unc-13 homolog B,and ZP4-regulating synaptic membrane exocytosis protein 1(RIMS1)showed the highest prediction value,and all these markers are involved in the acrosome reaction.Conclusion:The EQTN-ZP4 model was efficient in clustering the high-fertility group and may be useful for selection of animal that has superior fertility in the livestock industry.Compared to the EQTN-ZP4 model,the ZP4-RIMS1 model was more efficient in clustering the low-fertility group and may be useful in the diagnosis of male infertility in humans and other animals.The appointed translational animal model and established biomarker combination can be widely used in various scientific fields such as biomedical science.
基金This work was supported by the National Institutes of Health,National Institute of Neurological Disorders and Stroke grant R01NS093314 as well as Small Business Innovation Research grant 1R43NS103596-01.
文摘Magnetic resonance imaging(MRI)is a clinically relevant,real-time imaging modality that is frequently utilized to assess stroke type and severity.However,specific MRI biomarkers that can be used to predict long-term functional recovery are still a critical need.Consequently,the present study sought to examine the prognostic value of commonly utilized MRI parameters to predict functional outcomes in a porcine model of ischemic stroke.Stroke was induced via permanent middle cerebral artery occlusion.At 24 hours post-stroke,MRI analysis revealed focal ischemic lesions,decreased diffusivity,hemispheric swelling,and white matter degradation.Functional deficits including behavioral abnormalities in open field and novel object exploration as well as spatiotemporal gait impairments were observed at 4 weeks post-stroke.Gaussian graphical models identified specific MRI outputs and functional recovery variables,including white matter integrity and gait performance,that exhibited strong conditional dependencies.Canonical correlation analysis revealed a prognostic relationship between lesion volume and white matter integrity and novel object exploration and gait performance.Consequently,these analyses may also have the potential of predicting patient recovery at chronic time points as pigs and humans share many anatomical similarities(e.g.,white matter composition)that have proven to be critical in ischemic stroke pathophysiology.The study was approved by the University of Georgia(UGA)Institutional Animal Care and Use Committee(IACUC;Protocol Number:A2014-07-021-Y3-A11 and 2018-01-029-Y1-A5)on November 22,2017.
基金supported by the National Centre for Research and Development TECHMATSTRATEG-Ⅲ/0027/2019,POWR.03.02.00-00-I006/17the IDUB UAM。
文摘Meniscus injuries are widespread and the available treatments do not offer enough healing potential.Here,we provide critical support for using pigs as a biological model for meniscal degeneration and the development of cutting-edge therapies in orthopedics.We present a single-cell transcriptome atlas of the meniscus,consisting of cell clusters corresponding to four major cell types:chondrocytes,endothelial cells,smooth muscle cells,and immune cells.Five distinct chondrocyte subclusters(CH0–CH4)were annotated,of which only one was widespread in both the red and white zones,indicating a major difference in the cellular makeup of the zones.Subclusters distinct to the white zone appear responsible for cartilage-specific matrix deposition and protection against adverse microenvironmental factors,while those in the red zone exhibit characteristics of mesenchymal stem cells and are more likely to proliferate and migrate.Additionally,they induce remodeling actions in other chondrocyte subclusters and promote the proliferation and maturation of endothelial cells,inducing healing and vascularization processes.Considering that they have substantial remodeling capabilities,these subclusters should be of great interest for tissue engineering studies.We also show that the cellular makeup of the pig meniscus is comparable to that of humans,which supports the use of pigs as a model in orthopedic therapy development.
基金Supported by the Fundamental Research Funds for the Central Universities of Central South University,No.2024XQLH027.
文摘BACKGROUND Colonic anastomotic leakage(AL)remains a feared complication of colorectal surgery.Usually,a defunctioning stoma or a proximal colostomy is performed to reduce the AL rate but cannot completely prevent AL.Moreover,defunctioning colostomy is associated with high morbidity.This study assessed the feasibility of completely preventing colonic AL using total enteric flow diversion without a defunctioning stoma in a pig model of colonic AL.AIM To determine the feasibility of preventing colonic AL via total enteric flow diversion in pigs.METHODS A total of 14 pigs underwent surgery to create colon anaesthesia with severe defects for establishing the AL model.The pigs were then randomized into the control group(n=7),which received no further therapy,and a diversion group(n=7),which underwent placement of a modified ileostomy tube to divert the enteric contents from the colon externally.The general condition,serum Creactive protein level,white blood cell count,5-day incidence of colon AL,and development of abdominal abscesses were evaluated.RESULTS A modified ileostomy tube with a balloon was placed and pressurized to 20 kPa at a distance of 10-20 cm proximal to the ileocecal valve,effectively obstructing the intestine without causing injury and efficiently diverting the enteric contents.In the diversion group,no cases of peritonitis or abscess were observed.In contrast,all pigs in the control group developed either abdominal abscesses or peritonitis.CONCLUSION Instead of ileostomy or colostomy,the total enteric flow diversion technique with the placement of a modified ileostomy tube and balloon in the ileum can effectively or completely prevent colon AL.
基金study was supported by National Natural Science Foundation of China(31730091)the National Key Research and Development Program of China(2017YFD0500503).
文摘Background:Previous studies have shown that exogenous short-chain fatty acids(SCFAs)introduction attenuated the body fat deposition in conventional mice and pigs.However,limited studies have evaluated the effects of exogenously introduced SCFAs on the lipid and glucose metabolism independently of the gut microbiota.This study was to investigate the effects of exogenous introduction of SCFAs on the lipid and glucose metabolism in a germ-free(GF)pig model.Methods:Twelve hysterectomy-derived newborn pigs were reared in six sterile isolators.All pigs were hand-fed with sterile milk powder for 21 d,then the sterile feed was introduced to pigs for another 21 d.In the second 21-d period,six pigs were orally administrated with 25 mL/kg sterile saline per day and considered as the GF group,while the other six pigs were orally administrated with 25 mL/kg SCFAs mixture(acetic,propionic,and butyric acids,45,15,and 11 mmol/L,respectively)per day and regarded as FA group.Results:Orally administrated with SCFAs tended to increase the adiponectin concentration in serum,enhance the CPT-1 activity in longissimus dorsi,and upregulate the ANGPTL4 mRNA expression level in colon(P<0.10).Meanwhile,the mRNA abundances of ACC,FAS,and SREBP-1C in liver and CD36 in longissimus dorsi of the FA group were decreased(P<0.05)compared with those in the GF group.Besides,the mRNA expression of PGC-1αin liver and LPL in longissimus dorsi tended to(P<0.10)upregulate and downregulate respectively in the FA group.Moreover,oral administration of SCFAs tended to increase the protein level of GPR43(P<0.10)and decrease the protein level of ACC(P<0.10)in liver.Also,oral administration of SCFAs upregulated the p-AMPK/AMPK ratio and the mRNA expressions of GLUT-2 and GYS2 in liver(P<0.05).In addition,the metabolic pathway associated with the biosynthesis of unsaturated fatty acids was most significantly promoted(P<0.05)by oral administration of SCFAs.Conclusions:Exogenous introduction of SCFAs might attenuate the fat deposition and to some extent improve the glucose control in the pig model,which occurred independently of the gut microbiota.
基金supported by the National Key R&D Program of China(2021YFD1300403)the Major Program of Heilongjiang Province of China(2021ZX12B08-02).
文摘Background Cold regions have long autumn and winter seasons and low ambient temperatures.When pigs are unable to adjust to the cold,oxidative damage and inflammation may develop.However,the differences between cold and non-cold adaptation regarding glucose and lipid metabolism,gut microbiota and colonic mucosal immunological features in pigs are unknown.This study revealed the glucose and lipid metabolic responses and the dual role of gut microbiota in pigs during cold and non-cold adaptation.Moreover,the regulatory effects of dietary glucose supplements on glucose and lipid metabolism and the colonic mucosal barrier were evaluated in cold-exposed pigs.Results Cold and non-cold-adapted models were established by Min and Yorkshire pigs.Our results exhibited that cold exposure induced glucose overconsumption in non-cold-adapted pig models(Yorkshire pigs),decreasing plasma glucose concentrations.In this case,cold exposure enhanced the ATGL and CPT-1αexpression to promote liver lipolysis and fatty acid oxidation.Meanwhile,the two probiotics(Collinsella and Bifidobacterium)depletion and the enrichment of two pathogens(Sutterella and Escherichia-Shigella)in colonic microbiota are not conducive to colonic mucosal immunity.However,glucagon-mediated hepatic glycogenolysis in cold-adapted pig models(Min pigs)maintained the stability of glucose homeostasis during cold exposure.It contributed to the gut microbiota(including the enrichment of the Rikenellaceae RC9 gut group,[Eubacterium]coprostanoligenes group and WCHB1-41)that favored cold-adapted metabolism.Conclusions The results of both models indicate that the gut microbiota during cold adaptation contributes to the protection of the colonic mucosa.During non-cold adaptation,cold-induced glucose overconsumption promotes thermogenesis through lipolysis,but interferes with the gut microbiome and colonic mucosal immunity.Furthermore,glucagon-mediated hepatic glycogenolysis contributes to glucose homeostasis during cold exposure.
基金supported by Zhangjiakou Project of Science and Technology Studies and Development Planning(Grand No.1321078D)
文摘Objective: To investigate the electrophysiological changes of autonomic cells in left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with chronic heart failure.Methods: Guinea pigs model of iron deficiency anemia complicated with chronic heart failure in 10 guinea pigs of the experimental group was made by feeding a low iron diet,pure water and subcutaneous injection of isoproterenol. The control group consisting of 11 guinea pigs was given normal food, normal water and injected with normal saline. The left ventricular outflow tract model specimen was also prepared. The standard microelectrode technique was used to observe electrophysiological changes of autonomic cells in the outflow tract of left ventricular heart failure complicated with iron deficiency anemia in guinea pig model. The indicators of observation were maximal diastolic potential, action potential amplitude, 0 phase maximal depolarization velocity, 4 phase automatic depolarization velocity, repolarization 50% and 90%, and spontaneous discharge frequency.Results: Compared with the control group, 4 phase automatic depolarization velocity,spontaneous discharge frequency and 0 phase maximal depolarization velocity decreased significantly(P < 0.01) and action potential amplitude reduced(P < 0.01) in model group. Moreover, repolarization 50% and 90% increased(P < 0.01).Conclusions: There are electrophysiological abnormalities of the left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with heart failure.
文摘Functional magnetic resonance imaging(fMRI) is em-ployed in many behavior analysis studies, with blood oxygen level dependent-(BOLD-) contrast imaging being the main method used to generate images. The use of BOLD-contrast imaging in f MRI has been refined over the years, for example, the inclusion of a spin echo pulse and increased magnetic strength were shown to produce better recorded images. Taking careful precautions to control variables during measurement, comparisons between different specimen groups can be illustrated by f MRI imaging using both quantitative and qualitative methods. Differences have been observed in comparisons of active and resting, developing and aging, and defective and damaged brains in various studies. However, cognitive studies using f MRI still face a number of challenges in interpretation that can only be overcome by imaging large numbers of samples. Furthermore, f MRI studies of brain cancer, lesions and other brain pathologies of both humans and animals are still to be explored.
基金Supported by Qilu Hospital of Shandong University (No.201805049)
文摘AIM： To investigate the effects of green flickering light on refractive development and expression of muscarinic acetylcholine receptor（mAChR） M1 in the eyes of guinea pigs.METHODS： Thirty guinea pigs（15-20 days old） were randomly divided into three groups（n=10/group）. Animals in group I were raised in a completely closed carton with green flickering light illumination. Those in group II were kept in the open top closed carton under normal natural light. Guinea pigs were raised in a sight-widen cage under normal natural light in group III. The refractive status and axial length were measured before and after 8 weeks＇ illumination. Moreover, total RNA extracted from retinal, choroidal, and scleral tissues were determined by real-time reverse transcription polymerase chain reaction（RT-PCR）. The expressions of the receptor M1 were also explored in the retina, choroid, and sclera using immunohistochemistry.RESULTS： There was a remarkable reduction in refractive error and increase in axial length after 8-weeks＇ green flickering light stimulation（P〈0.001）. The expression of M1 receptor mRNA in sclera and retina in myopia group were remarkably lower than that in group II and III（P〈0.01）. Significant reduced expression of M1 receptor stimulated by green flickering light in retina and sclera tissues were also observed（P〈0.05）. However, there was no M1 receptor expression in choroid in 3 groups.CONCLUSION： Myopia can be induced by 8 weeks＇ green flickering light exposure in the animal model. M1 receptor may be involved causally or protectively in myopia development.
基金funded by the National Research Foundationof Korea (No. NRF-2017R1C1B5076493).
文摘To apply a new airway treatment to humans, preclinical studies in an appropriate animalmodel is needed. Canine, porcine and leporine tracheas have been employed as animal airwaystenosis models using various methods such as chemical caustic agents, laser, and electrocautery.However, existing models take a long time to develop (3- 8 weeks) and the mechanism of stenosisis different from that in humans. The aim of the present study was to establish a new and fasttracheal stenosis model in pigs using a combination of cuff overpressure intubation (COI) andelectrocautery. Fourteen pigs were divided into three groups: tracheal cautery (TC) group (n=3),COI group (n=3), and COI-TC combination group (n=8). Cuff overpressure (200/400/500 mmHg)was applied using a 9-mm endotracheal tube. Tracheal cautery (40/60 watts) was performed usinga rigid bronchoscopic electrocoagulator. After intervention, the pigs were observed for 3 weeks andbronchoscopy was performed every 7 days. When the cross-sectional area decreased by > 50%, itwas confirmed that tracheal stenosis was established. The time for tracheal stenosis was 14 days inthe TC group and 7 days in the COI-TC combination group. In the COI group, no stenosis occurred.In the COI-TC group, electrocautery (40 watts) immediately after intubation for>1 h with a cufpressure of 200 mmHg or more resulted in suficient tracheal stenosis within 7 days. Moreover, thedegree of tracheal stenosis increased in proportion to the cuff pressure and tracheal intubation time.The combined use of cuf overpressure and electrocautery helped to establish tracheal stenosis inpigs rapidly.
基金National Natural Science Foundation of China(31730091)the National Key Research and Development Program of China(2017YFD0500503).
文摘This study was conducted to investigate host-microbiota interactions and explore the effects of maternal gut microbiota transplantation on the growth and intestinal functions of newborns in a germ-free(GF)pig model.Twelve hysterectomy-derived GF Bama piglets were reared in 6 sterile isolators.Among them,6 were considered as the GF group,and the other 6 were orally inoculated with healthy sow fecal suspension as fecal microbiota transplanted(FMT)group.Another 6 piglets from natural birth were regarded as the conventional(CV)group.The GF and FMT groups were hand-fed with Co60-y-irradiated sterile milk powder,while the CV group was reared by lactating Bama sows.All groups were fed for 21 days.Then,all piglets and then were switched to sterile feed for another 21 days.Results showed that the growth performance,nutrient digestibility,and concentrations of short-chain fatty acids in the GF group decreased(P<0.05).Meanwhile,the serum urea nitrogen concentration and digesta pH values in the GF group increased compared with those in the FMT and CV groups(P<0.05).Compared with the CV group,the GF group demonstrated upregulation in the mRNA expression levels of intestinal barrier function-related genes in the small intestine(P<0.05).In addition,the mRNA abundances of intestinal development and absorption-related genes in the small intestine and colon were higher in the GF group than in the CV and FMT groups(P<0.05).The FMT group exhibited greater growth performance,lipase activity,and nutrient digestibility(P<0.05),higher mRNA expression levels of intestinal development and barrier-related genes in the small intestine(P<0.05),and lower mRNA abundances of pro-inflammatory factor in the colon and jejunum(P<0.05)than the CV group.In conclusion,the absence of gut microbes impaired the growth and nutrient digestibility,and healthy sow gut microbiota transplantation increased the growth and nutrient digestibility and improved the intestinal development and barrier function of newborn piglets,indicating the importance of intestinal microbes for intestinal development and functions.
基金the National Key Research and Development Program of China(2017YFA0105103,2021YFA0805903)the National Natural Science Foundation of China(81941004,32170542)+10 种基金2020 Research Program of Sanya Yazhou Bay Science and Technology City(202002011)Major Science and Technology Projects of Hainan Province(ZDKJ2021030)Key Research&Development Program of Hainan Province(ZDYF2021SHFZ052)Youth Innovation Promotion Association of the Chinese Academy of Sciences(2019347)Young Elite Scientist Sponsorship Program by CAST(YESS20200024)Biological Resources Progaramme,Chinese Academy of Sciences(KFJBRP-017-57)Key Research&Development Program of Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory)(2018GZR110104004)China Postdoctoral Science Foundation(2020M682943)Science and Technology Planning Project of Guangdong Province,China(2019A030317010,2020B1212060052,2021B1212040016,2021A1515011110)Science and Technology Program of Guangzhou,China(202007030003)Research Unit of Generation of Large Animal Disease Models,Chinese Academy of Medical Sciences(2019-I2M-5-025)。
文摘Inducible expression systems are indispensable for precise regulation and in-depth analysis of biological process.Binary Tet-On system has been widely employed to regulate transgenic expression by doxycycline.Previous pig models with tetracycline regulatory elements were generated through random integration.This process often resulted in uncertain expression and unpredictable phenotypes,thus hindering their applications.Here,by precise knock-in of binary Tet-On 3G elements into Rosa26 and Hipp11 locus,respectively,a double knock-in reporter pig model was generated.We characterized excellent properties of this system for controllable transgenic expression both in vitro and in vivo.Two att P sites were arranged to flank the td Tomato to switch reporter gene.Single or multiple gene replacement was efficiently and faithfully achieved in fetal fibroblasts and nuclear transfer embryos.To display the flexible application of this system,we generated a pig strain with Dox-inducing h KRASexpression through phiC31 integrase-mediated cassette exchange.After eight months of Dox administration,squamous cell carcinoma developed in the nose,mouth,and scrotum,which indicated this pig strain could serve as an ideal large animal model to study tumorigenesis.Overall,the established pig models with controllable and switchable transgene expression system will provide a facilitating platform for transgenic and biomedical research.
基金the Strategic Priority Research Programs of CAS(XDA16030300)the National Natural Science Foundation of China(81671274,31272440,and 31801031)+1 种基金the National Transgenic Project of China(2016ZX08009003-006-007)the Elite Youth Program of the Chinese Academy of Agricultural Sciences(ASTIP-IAS05).
文摘Harlequin ichthyosis (HI) is a severe genetic skin disorder and caused by mutation in the ATP-binding cassette A12 (ABCA12) gene. The retinoid administration has dramatically improved long-term survival of HI, but improvements are still needed. However, the ABCA12 null mice failed to respond to retinoid treatment, which impedes the development of novel cure strategies for HI. Here we generated an ethylnitrosourea mutagenic HI pig model (named Z9), which carries a novel deep intronic mutation IVS49-727 A>G in the ABCA12 gene, resulting in abnormal mRNA splicing and truncated protein production. Z9 pigs exhibit significant clinical symptom as human patients with HI. Most importantly, systemic retinoid treatment significantly prolonged the life span of the mutant pigs via improving epidermal maturation, decreasing epidermal apoptosis, and triggering the expression of ABCA6. Taken together, this pig model perfectly resembles the clinical symptom and molecular pathology of patients with HI and will be useful for understanding mechanistic insight and developing therapeutic strategies.
基金supported the Natural Science Foundation of Jilin Province(YDZJ202201ZYTS101).
文摘In a recent study,Yong-Guang Yang and colleagues developed a severely immunodeficient pig model that supports long-term engraftment and multilineage differentiation of human hematopoietic stem/progenitor cells(HSPCs).1 This breakthrough addresses key limitations of current humanized mouse models,including their small size and limited lifespan,and offers a promising platform for the large-scale production of human immune cells,potentially functioning as an in vivo bioreactor.
基金supported by the National Natural Science Foundation of China(31972582)the STI 2030-Major Project(2023ZD04072)+4 种基金the Major Project of Changsha City(kh2401015)the Science and Technology Innovation Program of Hunan Province(2021RC4039)the Key Research and Development Program of Hunan Province(2022NK2026)the Youth Innovation Promotion Association CAS(Y202079)China Agriculture Research System of MOF and MARA(CARS-35)。
文摘Metabolites and metabolism-related gene expression profiles in skeletal muscle change dramatically under obesity,aging and metabolic disease.Since obese and lean pigs are ideal models for metabolic research.Here,we compared metabolome and transcriptome of Longissimus dorsi(LD)muscle between Taoyuan black(TB,obese)and Duroc(lean)pigs at different ages.We defined the“window phase”of intramuscular fat(IMF)deposition in TB pig,which has significantly higher IMF than Duroc pig.Our results displayed discrepant lipid composition and different expression genes(DEGs)enriched in lipid metabolism,and both metabolome and transcriptome analyses revealed stronger energy expenditure and more active amino acid and protein metabolism in Duroc pig.10 up-and 51 down-regulated biomarker metabolites with age-and breed-specificity were identified.Potential secretory metabolites,including organic acid(fumaric acid,succinate,malic acid,and gamma-aminobutyric acid),amino acid(L-lysine,and L-glutamic acid),lipid(2-hydroxyisovaleric acid,and L-carnitine)were demonstrated a significant correlation with IMF deposition.Our research highlights the huge difference of metabolic spectrum in skeletal muscle between obese and lean model and muscle-derived secretory metabolites might act as an ambassador of intercellular communication to regulate systematic metabolism.
基金National Natural Science Foundation of China(grant no.82271329grant no.82471356).
文摘Background and purpose The low-field MRI is a promising tool to accurately diagnose strokes.We here report our study on the accuracy of a 0.23-Tesla(0.23-T)MRI using the haematoma enhanced inversion recovery(HEIR)sequence to detect acute ischaemic stroke(AIS)and intracerebral haemorrhage(ICH)within 24 hours of symptom onset.Methods A novel HEIR sequence based on fluid-attenuated inversion recovery T1-weighted,with a scanning time of 1 min and 17 s,was developed using an ICH and AIS pig model on a 0.23-T MRI.Images of the pig model were obtained hourly for 24 hours in order to monitor value changes on T1/T2 and verify the differential diagnosis of AIS and ICH.Then,30 patients with AIS and 30 patients with ICH with confirmed diagnoses by 3T-MRI/CT were included.Diagnostic criteria on a 0.23-T MRI for ICH was the hyperintensity signal on both the diffusion-weighted imaging(DWI)and HEIR sequence,while for AIS was the hyperintensity on DWI and isointensity on the HEIR sequence.Two blinded raters independently assessed the images obtained by the 0.23-T MRI for the presence of ICH/AIS.Results In the pig model,setting the inversion time to 800 ms enabled clear differentiation of ICH from brain parenchymal tissue and AIS.In real patients,a correct 0.23-T MRI diagnosis of either an AIS or ICH was made in all 60 patients within 24 hours of symptom onset(100%overall accuracy).No adverse events occurred.Conclusions The 0.23-T MRI may have the potential to differentiate cerebral haemorrhage from cerebral infarction with both speed and accuracy,making brain MRI scans easier,faster and cheaper.It might be possible to improve the screening imaging process for strokes in the emergency room.Further multicentre studies are needed to validate our findings.
