Minipig Research Articles

A modified chain-based sponge dressing controls junctional hemorrhage in the tactical combat casualty care simulation of pigs

BackgroundHemorrhage has always been the focus of battlefield and pre-hospitalization treatment. With the increasing fatality rates associated with junctional bleeding, treatment of bleeding at junctional sites has gradually gained attention in battlefield trauma emergency care. We designed a modified chain-based sponge dressing with a medical polyvinyl alcohol sponge that can be used to treat junctional hemorrhage and tested its hemostatic efficacy and biocompatibility.MethodsTwenty adult Bama miniature pigs were randomly divided into the modified chain-based sponge dressing (MCSD) and standard gauze (SG) groups. The right femoral artery of the pigs was shot at after anesthesia. The Bama miniature pigs were moved to the safety zone immediately to assess the condition according to the MARCH strategy, which evaluates massive hemorrhaging, airway obstruction, respiratory status, circulatory status, head injury & hypothermia. Hemoglobin and coagulation status were checked during the experiment.Among the pigs in which the inguinal hemorrhagic model based on bullet penetrating wounds was successfully established, those in the MCSD group received a disinfected MCSD for hemostasis, while those in the SG group received standard gauze in an imbricate manner to pack the bullet exit and entrance wounds to stop bleeding until the wound was filled, followed by compression for 3 min at sufficient pressure. CT scanning, transmission electron microscopy, and HE staining were conducted after experiment.ResultsThe MCSD group showed lower hemostasis time and blood loss than the gauze group. The MCSD group also showed a higher success rate of treatment,more stable vital signs and hemoglobin level. The CT scanning results showed tighter packing without large gaps in the MCSD group. The histopathological assessments and the transmission electron microscopy and HE staining findings indicated good biocompatibility of the polyvinyl alcohol sponge.ConclusionThe MCSD met the battlefield’s requirements of speedy hemostasis and biosafety for junctional hemorrhage in Bama miniature pigs. Moreover, in comparison with the conventional approach for hemostasis, it showed more stable performance for deep wound hemostasis. These findings provide the theoretical and experimental basis for the application of MCSD in the treatment of hemorrhage in the battlefield in the future.

Abstract 15329: Echocardiographic Assessment of Cardiac Function in Preterm Miniature Pigs Supported With a Pumped Artificial Placenta

Introduction: Preterm birth is the leading cause of infant mortality and morbidity. Artificial placenta (AP) technology aims to support premature fetal piglets in an environment simulating intrauterine physiology to maintain fetal circulation and improve outcomes and survival. Hypothesis: We sought to understand the fetal cardiovascular physiology and mode of cardiac failure associated with a pumped extracorporeal membrane oxygenation circuit connected to the fetal circulation. Methods: Following umbilical cannulation, connection to a centrifugal pump and neonatal oxygenator and transition of preterm Yucatan miniature piglets to a fluid-filled biobag, echocardiographic studies measured ventricular function and fluid status including Dopplers, and strain analysis both prenatally and while maintained on the AP circuit. Results: AP fetuses (n=13; GA 102±4d; 616±139g; survival 46.4±46.8h) were tachycardic and hypertensive with supraphysiologic circuit flows initially. Strain analysis revealed significantly reduced RV strain rate within the first five hours of AP support compared to paired prenatal measurements (p=0.005) . Fetuses supported for <24h had significantly lower overall RV global longitudinal strain (p=0.001) than those surviving >24h. M-mode imaging revealed progressive cardiac hypertrophy with an increase in interventricular septum (p=0.03) and LV free (p=0.04) wall thickness from first to last measurements. Doppler measurements showed elevated peak flow velocity patterns in the MCA and reductions in MCA-PI from start to end of support (p=0.04) as well as compared to unpaired prenatal controls (p=0.03, p=0.001) . Imaging findings included ascites, skin edema, and dilated hepatic veins. Conclusions: AP fetuses exhibited signs of excessive volume and pressure loading resulting in impaired contractility, including supraphysiologic umbilical flows, reduced RV strain and strain rate, and evidence of elevated filling pressures. Myocardial hypertrophy and changes in cerebral perfusion indicated adaptation to increased afterload, potentially affecting cardiovascular and cerebrovascular development and leading to fetal deterioration. Further advancements in AP technology are necessary before clinical translation.

Abstract 15472: The IRIS™ Valve: A Growth Accommodating Transcatheter Pulmonary Valve for Pediatric Patients

Introduction: It is estimated that at least 1 million children are living with a congenital heart defect (CHD) in the U.S. Most of these patients are born with some degree of right ventricular outflow tract (RVOT) abnormalities that at some point require pulmonary valve replacement to mitigate the detrimental effects of pulmonary valve regurgitation (PVR) on the right ventricle (RV). Due to their size, available solutions, e.g., the Melody™ valve (Medtronic Inc., Minneapolis, MN) cannot be implanted in small children under 20 Kg. Those must wait until they grow to a suitable weight to receive the valve, which may lead to RV dysfunction. Hypothesis: To mitiagte the detrminental effects to the RV, the IRIS™ valve is implantable at 12 mm with a 12-Fr delivery catheter. As the valve annulus grows, the valve would be balloon expanded up to 20 mm. Methods: The IRIS™ valve has been designed according to origami concepts such that its three leaflets would retain a fully-coapted form as it is expanded to a larger size. Here we report our methods and results of implantation in 9-17Kg Yucatan pigs. Results: The IRIS™ valve is made of thin porcine pericardial leaflets sewn into a laser-cut stainless-steel stent. The stented valve is covered with an ePTFE skirt to mitigate paravalvular leak. The IRIS™ valve is fully crimped over a 12-Fr delivery catheter for transfemoral implantation. We have successfully implanted the valve at the native pulmonary valve position with minimal regurgitation in three pigs ranging from 9 to 17kg with varying pulmonary annulus sizes. Conclusions: The IRIS™ transcatheter pulmonary valve has been successfully developed according to proprietary origami concepts. Using the 12Fr delivery catheter, the smallest delivery profile reported, we successfully implanted the IRIS™ valve within the pulmonary annulus of mini pigs with encouraging results.

Mono- and multimeric PSMA-targeting small molecule-thorium-227 conjugates for optimized efficacy and biodistribution in preclinical models

PurposePSMA (prostate-specific membrane antigen) is highly expressed on prostate cancer (PrCa) cells and extensively used as a homing target for PrCa treatment. Most prominently, PSMA-targeting conjugate PSMA-617, carrying a DOTA chelator and labeled with therapeutic radionuclides like beta-emitting lutetium-177 or alpha-emitting actinium-225, has shown clinical activity in PrCa patients. We sought to develop PSMA-targeting small molecule (SMOL) conjugates that show high uptake in PSMA-expressing tumors and fast clearance, and can easily be labeled with the alpha emitter thorium-227 (half-life 18.7 days).MethodsA novel linker motif with improved competition against 3H-PSMA-617 on PSMA-expressing LNCaP cells was identified. A 2,3-hydroxypyridinone chelator modified with carboxyl groups (carboxy-HOPO) with increased hydrophilicity and robust labeling with thorium-227 was developed and allowed the synthesis of mono-, di-, tri-, and tetrameric conjugates. The resulting monomeric and multimeric PSMA SMOL-TTCs (targeted thorium conjugate) were evaluated for cellular binding, internalization, and antiproliferative activity. The in vivo antitumor efficacy of the PSMA SMOL-TTCs was determined in ST1273 and KUCaP-1 PrCa models in mice, and their biodistribution was assessed in cynomolgus monkeys, minipigs, and mice.ResultsThe monomeric and multimeric PSMA SMOL conjugates were readily labeled with thorium-227 at room temperature and possessed high stability and good binding, internalization, and antiproliferative activity in vitro. In vivo, the monomeric, dimeric, and trimeric PSMA SMOL-TTCs showed fast clearance, potent antitumor efficacy, and high uptake and retention in prostate tumors in mice. No major uptake or retention in other organs was observed beyond kidneys. Low uptake of free thorium-227 into bone confirmed high complex stability in vivo. Salivary gland uptake remained inconclusive as mini pigs were devalidated as a relevant model and imaging controls failed in cynomolgus monkeys.ConclusionMonomeric and multimeric PSMA SMOL-TTCs show high tumor uptake and fast clearance in preclinical models and warrant further therapeutic exploration.

AKT from dental epithelium to papilla promotes odontoblast differentiation

Epithelial–mesenchymal interactions occur during tooth development. The dental epithelium (DE) is regarded as the signal center that regulates tooth morphology. However, the mechanism by which DE regulates the differentiation of mesenchyme-derived dental papilla (DP) into odontoblasts remains unclear. Using miniature pigs as a model, we analyzed the expression profiles of the DE and DP during odontoblast differentiation using high-throughput RNA sequencing. The phosphatidylinositol-3-kinase (PI3K)/AKT pathway is one of the most enriched pathways in both DE and DP. The PI3K/AKT pathway was first activated in the inner enamel epithelium but not in the DP on embryonic day 50. This pathway was then activated in the odontoblast layer on embryonic day 60. We showed that AKT activation promoted odontoblast differentiation of DP cells. We further demonstrated that activation of PI3K/AKT signaling in the DE effectively increased the expression levels of AKT and dentin sialophosphoprotein in DP cells. Additionally, we found that DE cells secreted collagen type IV alpha 6 chain (COL4A6) downstream of epithelial AKT signaling to positively regulate mesenchymal AKT levels. Therefore, our data suggest that PI3K/AKT signaling from the DE to the DP promotes odontoblast differentiation via COL4A6 secretion.

Integration of 16S rRNA gene sequencing and LC/MS-based metabolomic analysis of early biomarkers of acute ischaemic stroke in Tibetan miniature pigs

Acute ischaemic stroke (AIS) is a complex, systemic, pathological, and physiological process. Systemic inflammatory responses and disorders of the gut microbiome contribute to increased mortality and disability following AIS. We conducted 16S high-throughput sequencing and ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry-based non-targeted metabolomic analyses of the plasma from a Tibetan miniature pig middle cerebral artery occlusion (MCAO) model. A significant decrease in the abundance of Firmicutes and a significant increase in the abundance of Actinobacteria were observed after the onset of AIS. Among the plasma metabolites, the levels of phospholipids and amino acids were considerably altered. Loading values and differential metabolite–bacterial group association analyses of the metabolome and microbiome indicated a correlation between the microbiome and metabolome of Tibetan miniature pigs after MCAO. Furthermore, significant changes were observed in the ABC transporter pathway and purine metabolism in the gut microbiome–plasma metabolome during the early stage of AIS. Kyoto Encyclopaedia of Genes and Genomes enrichment analysis showed that arginine, proline, and cyanoamino acid metabolism was upregulated while ABC transporter metabolism pathway and carbohydrate digestion and absorption were substantially downregulated. The results of this study suggest that AIS affects the gut microbiota and plasma metabolites in Tibetan miniature pigs and that faecal microbiota transplantation could be a potential therapeutic approach for AIS.

Benzbromarone Induces Targeted Degradation of HSP47 Protein and Improves Hypertrophic Scar Formation

Fibrotic diseases are characterized by the abnormal accumulation of collagen in the extracellular matrix (ECM), leading to the functional impairment of various organs. In the skin, excessive collagen deposition manifests as hypertrophic scars and keloids, placing a substantial burden on patients and the health care system worldwide. Heat shock protein 47 (HSP47) is essential for proper collagen assembly and contributes to fibrosis. However, identifying clinically applicable HSP47 inhibitors has been a major pharmaceutical challenge. Here, we identified benzbromarone (BBR) as a HSP47 inhibitor for hypertrophic scarring treatment. BBR inhibited collagen production and secretion in fibroblasts from keloid patients by binding to HSP47 and inhibiting the interaction between HSP47 and collagen. Interestingly, BBR not only inhibits HSP47 but also acts as a molecular glue degrader that promotes its proteasome-dependent degradation. Through these molecular mechanisms, BBR effectively reduced hypertrophic scarring in mini pigs and mice with burns and/or excisional skin damage. Thus, these findings suggest that BBR can be used to clinically treat hypertrophic scars and, more generally, fibrotic diseases.

The New Quantum Image by Dynamic Nuclear Polarized MRI for the Assessment of Cardiac Radioablation to the Cavotricuspid Isthmus

Cardiac arrhythmias are usually treated with invasive, time consuming catheter ablation techniques. While recently stereotactic body radiotherapy (SBRT) is an emerging non-invasive treatment in the management of cardiac arrhythmias. To identify and assess the cardiac radioablation by MR examination, including diffusion-weighted MRI, dynamic Gd-enhanced MRI, MR spectroscopy, and T2-weighted MRI early after SBRT is very difficult. We have been developing the free radical imaging methods using Dynamic Nuclear Polarization (DNP)-MRI with nitroxyl radicals as a redox probe (e.g., 4-Methacryloyloxy-2,2,6,6-tetramethylpiperidine-1-oxyl (Tempo methacrylate; TempoMC)). In this study, we examined the possibility of in vivo spatiotemporal visualization of SBRT for cardiac radioablation to the cavotricuspid isthmus (CTI) based on redox reaction by in vivo DNP-MRI. All animal procedures were approved by institutional animal care and use committee and performed in full compliance with its guidelines. This study was conducted with two approaches. First, four mini pigs underwent electrophysiology assessment using electroanatomic mapping (EAM) before and 3 months after SBRT with single-fraction doses of 25 Gy. The target of CTI was defined by cardiologist. We defined the planning target volume (PTV): the internal margin (IM) + set up margin (SM) = SI 15 mm, AP 10mm, LR 10 mm were added to the target. Radiotherapy plans were created by the software used in daily clinical practice. Second, free radical imaging by low filed type of DNP-MRI was performed on the four mice before and after 25 Gy and 10 Gy irradiation to whole heart. ESR signal measurements were also performed. A total dose of 25 Gy was successfully delivered to PTV in a single procedure in all mini pigs. EAM visualized the irradiated site and confirmed clockwise conduction block across the CTI. Although routine MRI could not detect the cardiac radiation injury clearly, the four mice of heart were well delineated on MRI and clearly visualized by DNP-MRI. DNP-MRI signal of TempoMC were decreased depending on prescribed irradiation dose. These data demonstrated the safety and feasibility of SBRT for creating conduction block across the CTI in mini pigs. Although to identify and assess the irradiated site by routine MR examination was impossible, the free radical imaging methods using Dynamic Nuclear Polarization (DNP)-MRI with TempoMC could be a promising successful method for the assessment of cardiac radioablation. This new quantum image by DNP-MRI will open the possibility of treating cardiac arrhythmias by SBRT safety and noninvasively.

The Effects of an Osteoarthritic Joint Environment on ACL Damage and Degeneration: A Yucatan Miniature Pig Model.

Posttraumatic osteoarthritis (PTOA) arises secondary to joint injuries and is characteristically driven by inflammatory mediators. PTOA is often studied in the setting of ACL tears. However, a wide range of other injuries also lead to PTOA pathogenesis. The purpose of this study was to characterize the morphological changes in the uninjured ACL in a PTOA inflammatory environment. We retrospectively reviewed 14 ACLs from 13 Yucatan minipigs, 7 of which had undergone our modified intra-articular drilling (mIAD) procedure, which induced PTOA through inflammatory mediators. Seven ACLs were harvested from mIAD minipigs (PTOA) and seven ACLs from control minipigs with no cartilage degeneration (non-PTOA). ACL degeneration was evaluated using histological scoring systems. IL-1β, NF-κB, and TNF-α mRNA expression in the synovium was measured using qRT-PCR. PTOA minipigs demonstrated significant ACL degeneration, marked by a disorganized extracellular matrix, increased vascularity, and changes in cellular shape, density, and alignment. Furthermore, IL-1β, NF-κB, and TNF-α expression was elevated in the synovium of PTOA minipigs. These findings demonstrate the potential for ACL degeneration in a PTOA environment and emphasize the need for anti-inflammatory disease-modifying therapies following joint injury.

Shuangshen Ningxin capsule alleviates myocardial ischemia–reperfusion injury in miniature pigs by modulating mitophagy: network pharmacology and experiments in vivo

BackgroundMyocardial ischemia/reperfusion injury (MI/RI) is involved in a variety of pathological states for which there is no effective treatment exists. Shuangshen Ningxin (SSNX) capsule which is developed by Xiyuan Hospital, Chinese Academy of Traditional Chinese Medicine has been demonstrated to alleviate MI/RI, but its mechanism remains to be further elucidated.MethodsThe MI/RI miniature pigs model was constructed to assess the pharmacodynamics of SSNX by blocking the proximal blood flow of the left anterior descending branch of the cardiac coronary artery through an interventional balloon. The principal chemical compounds and potential targets of SSNX were screened by HPLC–MS and SwissTargetPrediction. The targets of MI/RI were identified based on Online Mendelian Inheritance in Man (OMIM) and GeneCards. Cytoscape 3.9.0 was applied to construct a protein–protein interaction (PPI) network, and Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed using metascape. To further validate the mechanism of SSNX, Molecular docking, Transmission electron microscopy, and Western blot analysis were used to test the effectiveness of targets in related pathways.ResultsOur results indicated that SSNX significantly improved cardiac function, attenuated myocardial I/R injury. Through network analysis, a total of 15 active components and 201 targets were obtained from SSNX, 75 of which are potential targets for the treatment of MI/RI. KEGG and MCODE analysis showed that SSNX is involved in the mitophagy signaling pathway, and ginsenoside Rg1, ginsenoside Rb1 and ginsenoside Rb2 are key components associated with the mitophagy. Further experimental results proved that SSNX protected mitochondrial structure and function, and significantly reduced the expression of mitophagy-related proteins PTEN-induced putative kinase 1 (PINK1), Parkin, FUN14 domain containing 1 (FUNDC1) and Bcl-2/E1B-19 kDa interacting protein 3 (BNIP3) in MI/RI miniature pigs.ConclusionIn our study, the integration of network pharmacology and experiments in vivo demonstrated that SSNX interfered with MI/RI by inhibiting mitophagy.

Ultrathin endoscope equipped with ultrasonic miniprobe for upper GI US in a porcine model

BackgroundUltrathin esophagogastroduodenoscopy (UT-EGD) is an ideal tool for unsedated upper gastrointestinal examination and pediatric gastroenterology but is rarely competent for endoscopic ultrasonography mini probe (EUS-MP). We developed a UT-EGD ultrasonography method(UT-EUS) and verified its clinical application value through animal experiments. MethodsFive Bama miniature pigs were selected. Using an acoustic medium, duodenum, stomach, and esophagus ultrasonography was performed respectively with conventional 20-MHz Endoscopic ultrasound miniprobe(EUS-MP-20), 20-MHz UT-EUS,(UT-EUS-20) and 30-MHz UT-EUS(UT-EUS-30). Record the time to acquire 5 consecutive stable ultrasound images, the number of identifiable wall layers, the quality, and the penetration depth of the images. ResultsThere was no significant difference in the time required to obtain images between EUS-MP-20 and UT-EUS-20 at each site (P > 0.05). UT-EUS-30 resolved the wall to more levels than UT-EUS-20 (P < 0.05). There was no significant difference between EUS-MP-20 and UT-EUS-20 in imaging quality and penetration depth (P > 0.05). ConclusionThe UT-EUS is easy to use, the obtained image quality is satisfactory, and it has potential clinical application value.

Removal of calciprotein particles from the blood using an adsorption column improves prognosis of hemodialysis miniature pigs

Hyperphosphatemia is a major risk for poor prognosis in patients with end-stage renal disease. However, the molecular mechanism behind this link remains elusive. We and others have demonstrated that serum phosphorus levels correlate positively with circulating levels of calciprotein particles (CPPs). CPPs are colloidal mineral-protein complexes containing insoluble calcium-phosphate precipitates and have been reported to induce calcification in cultured vascular smooth muscle cells and inflammatory responses in cultured macrophages. Hence, we hypothesize that CPPs may be responsible for disorders associated with hyperphosphatemia. Using hyperphosphatemic miniature pigs receiving hemodialysis, here we show that removal of CPPs from the blood with a newly developed CPP adsorption column improves survival and alleviates complications including coronary artery calcification, vascular endothelial dysfunction, metastatic pulmonary calcification, left ventricular hypertrophy, and chronic inflammation. The present study identifies CPPs as an effective therapeutic target and justifies clinical trials to determine whether the CPP adsorption column may be useful as a medical device for improving clinical outcomes of hemodialysis patients.

Infection of Feral Phenotype Swine with Japanese Encephalitis Virus.

Background: Japanese encephalitis virus (JEV) is a mosquito-borne zoonotic flavivirus and the leading cause of pediatric encephalitis in the Asian Pacific region. The transmission cycle primarily involves Culex spp. mosquitoes and Ardeid birds, with domestic pigs (Sus scrofa domestica) being the source of infectious viruses for the spillover of JEV from the natural endemic transmission cycle into the human population. Although many studies have concluded that domestic pigs play an important role in the transmission cycle of JEV, and infection of humans, the role of feral pigs in the transmission of JEV remains unclear. Since domestic and feral pigs are the same species, and because feral pig populations in the United States are increasing and expanding geographically, the current study aimed to test the hypothesis that if JEV were introduced into the United States, feral pigs might play a role in the transmission cycle. Materials and Methods: Sinclair miniature pigs, that exhibit the feral phenotype, were intradermally inoculated with JEV genotype Ib. These pigs were derived from crossing miniature domestic pig with four strains of feral pigs and were used since obtaining feral swine was not possible. Results: The Sinclair miniature pigs became viremic and displayed pathological outcomes similar to those observed in domestic swine. Conclusion: Based on these findings, we conclude that in the event of JEV being introduced into the United States, feral pig populations could contribute to establishment and maintenance of a transmission cycle of JEV and could lead to the virus becoming endemic in the United States.

Reconstruction of the temporomandibular joint using a vascularized medial femoral condyle osteocartilaginous flap: an experimental investigation in miniature pigs

BackgroundReconstruction of the temporomandibular joint (TMJ) is a significant challenge in maxillofacial surgery. A vascularized medial femoral condyle (MFC) osteocartilaginous flap is a good choice for TMJ reconstruction. In this study, we evaluated the radiographic and histological changes of MFC after TMJ reconstruction.MethodsA ramus-condyle unit (RCU) defect was created unilaterally in five adult male Bama miniature pigs. The ipsilateral vascularized MFC osteocartilaginous flap was used to reconstruct the TMJ, and the non-operative sides served as controls. Multislice spiral computed tomography (CT) was performed preoperatively, immediately postoperatively, and at two weeks, three months, and six months postoperatively. Three animals were euthanized at 6 months postoperatively. Their reconstructed condyles, natural condyles and the MFCs on the opposite side were collected and subjected to µCT and histological evaluation.ResultsIn the miniature pigs, the vascularized MFC osteocartilaginous flap was fused to the mandible, thus restoring the structure and function of the RCU. The postoperative radiographic changes and histological results showed that the reconstructed condyle was remodeled toward the natural condyle, forming a similar structure, which was significantly different from the MFC.ConclusionsIn miniature pigs, the RCU can be successfully reconstructed by vascularized osteocartilaginous MFC flap. The reconstructed condyle had almost the same appearance and histological characteristics as the natural condyle.

Experimental investigation of biostimulatory effects after polydioxanone thread insertion in a pig model.

Polydioxanone (PDO) threads have been widely used to tighten and lift the facial soft tissue. This research aims to determine the collagenation and inflammation changes that occur in the adipose tissue over time when different types of threads are implanted. Three threads types, PDO, poly glycolic-co-lactic acid (PGLA), and nylon, were inserted in the subcutaneous fat of 12-month-old Bama miniature pigs. Collagen production and inflammatory response were evaluated by hematoxylin and eosin and Masson trichrome staining at 1, 4, 12, 24, and 48 weeks. The integrity of the PDO thread lasted up to 24 weeks with mild inflammation and collagen production. The PGLA thread integrity lasted until 12 weeks and had a strong inflammatory response. The nylon thread's integrity was maintained for 48 weeks and showed minor inflammation and collagen production. Our data suggest that PDO thread is the best choice for clinicians, as it has a mild action process with minimal irritation, moderate collagen production, a reasonable explanation time, with obvious bridging fibrous tissue, and thickening action for the superficial fascia.

Metabolomics analyses of serum metabolites perturbations associated with Naja atra bite.

Naja atra bite is one of the most common severe snakebites in emergency departments. Unfortunately, the pathophysiological changes caused by Naja atra bite are unclear due to the lack of good animal models. In this study, an animal model of Naja atra bite in Guangxi Bama miniature pigs was established by intramuscular injection at 2 mg/kg of Naja atra venom, and serum metabolites were systematically analyzed using untargeted metabolomic and targeted metabolomic approaches. Untargeted metabolomic analysis revealed that 5045 chromatographic peaks were obtained in ESI+ and 3871 chromatographic peaks were obtained in ESI-. Screening in ESI+ modes and ESI- modes identified 22 and 36 differential metabolites compared to controls. The presence of 8 core metabolites of glutamine, arginine, proline, leucine, phenylalanine, inosine, thymidine and hippuric acid in the process of Naja atra bite was verified by targeted metabolomics significant difference (P<0.05). At the same time, during the verification process of the serum clinical samples with Naja atra bite, we found that the contents of three metabolites of proline, phenylalanine and inosine in the serum of the patients were significantly different from those of the normal human serum (P<0.05). By conducting functional analysis of core and metabolic pathway analysis, we revealed a potential correlation between changes in key metabolites after the Naja atra bite and the resulting pathophysiological alterations, and our research aims to establish a theoretical foundation for the prompt diagnosis and treatment of Naja atra bite.

Spatiotemporal expression of fibroblast growth factor 4 and 10 during the morphogenesis of deciduous molars in miniature pigs

ObjectiveFibroblast growth factors (FGFs) play pivotal roles in mediating interactions between dental epithelium and mesenchyme throughout tooth initiation and morphogenesis. This study aimed to elucidate the roles of FGF4 and FGF10 in the regulation of tooth development. DesignIn this study, we investigated spatiotemporal expression patterns of FGF4 and FGF10 in the third deciduous molars (DM3) of miniature pigs at the cap, early bell, and late bell stages. Pregnant miniature pigs were obtained, and the samples were processed for histological staining. Non-radioactive in situ hybridization, immunohistochemistry, and real-time PCR were used to detect mRNA and protein expression levels of FGF4 and FGF10. ResultsFGF4 was expressed in the dental epithelium and mesenchyme at the cap stage. At the early bell stage, epithelial expression of FGF4 was reduced while mesenchymal expression got stronger. At the late bell stage, the FGF4 expression was restricted to the inner enamel epithelium (IEE) and differentiating odontoblasts. FGF10 was expressed intensely in both epithelium and mesenchyme at the cap stage. The expression of FGF10 was concentrated in the secondary enamel knots and surrounding mesenchyme at the early bell stage. FGF10 was weakly detected in the IEE by the late bell stage. ConclusionsOur results indicated that FGF4 and FGF10 might have partially redundant functions in regulating epithelium morphogenesis. FGF4 may be involved in regulatory signaling cascades mediating interactions between the epithelium and mesenchyme. In addition, the downregulation of FGF10 expression may be associated with the cessation of mesenchymal cell proliferation and initiation of preodontoblast polarization.

Ethological Basis for the Relationship between Humans and Miniature Pigs

Ethological Basis for the Relationship between Humans and Miniature Pigs

Novel patch biomaterial treatment for colon diverticulosis in swine model.

Current leading managements for diverticular disease cannot prevent the recurrence of diverticulitis, bleeding and/or other complications. There is an immediate need for developing new minimal invasive therapeutic strategies to prevent and treat this disease. Through a biomechanical analysis of porcine colon with diverticular lesions, we proposed a novel adhesive patch concept aiming at mechanical reconstruction of the diseased colon wall. This study aims to evaluate the surgical feasibility (safety and efficacy) of pulmonary visceral pleura (PVP) patch therapy using a pig model of diverticulosis. Six female Yucatan miniature pigs underwent collagenase injection (CI) for the development of diverticular lesions. The lesions in each animal either received patch implantation (treated group, n = 40 for 6 pigs) or left intact (untreated group, n = 44 for 6 pigs). The normal colonic wall in each animal received patch implantation at two spots to serve as control (n = 12 for 6 pigs). After 3months of observation, the performance and safety of the patch treatment were evaluated through macroscopic and histological examination. We found that 95% of pouch-like herniation of the mucosa was prevented from the colon wall with the treatment. The pouch diameter was significantly reduced in the treated group as compared to the untreated group (p < 0.001). The patch application caused a significant increase in the levels of collagen of the colon tissue as compared to the untreated and control groups (p < 0.001). No difference was found in the lymphocyte and macrophage inflammatory infiltrate between the groups. Our results suggest that patch treatment efficiently inhibits the diverticular pouch deformation and promotes the healing of the colon wall with a normal inflammatory response, which may minimize the risk of diverticulosis reoccurrence and complications over time.

A TLR7/8 agonist increases efficacy of anti-fentanyl vaccines in rodent and porcine models.

Opioid use disorders (OUD) and overdose are public health threats worldwide. Widespread access to highly potent illicit synthetic opioids such as fentanyl is driving the recent rise in fatal overdoses. Vaccines containing fentanyl-based haptens conjugated to immunogenic carrier proteins offer a long-lasting, safe, and cost-effective strategy to protect individuals from overdose upon accidental or deliberate exposure to fentanyl and its analogs. Prophylactic or therapeutic active immunization with an anti-fentanyl vaccine induces the production of fentanyl-specific antibodies that bind the drug in the blood and prevent its distribution to the brain, which reduces its reinforcing effects and attenuates respiratory depression and bradycardia. To increase the efficacy of a lead anti-fentanyl vaccine, this study tested whether the incorporation of synthetic toll-like receptor (TLR) 4 and TLR7/8 agonists as vaccine adjuvants would increase vaccine efficacy against fentanyl challenge, overdose, and self-administration in either rats or Hanford miniature pigs. Formulation of the vaccine with a nucleolipid TLR7/8 agonist enhanced its immunogenicity and efficacy in preventing fentanyl-induced respiratory depression, analgesia, bradycardia, and self-administration in either rats or mini-pigs. These studies support the use of TLR7/8 adjuvants in vaccine formulations to improve their clinical efficacy against OUD and potentially other substance use disorders (SUD).