Experimental Animals Research Articles

PFKM Promotes the Progression of Gastric Cancer by Up-regulating CNTN1 Expression Through H3K18la Modification.

Gastric cancer (GC) stands as one of the most common malignancies globally, characterized by significant incidence rates. Phosphofructokinase muscle isoform (PFKM), a critical rate-limiting enzyme in glycolysis, has its expression modulated by lactate production in tumor cells. The objective of this study is to elucidate the underlying molecular mechanisms by which PFKM contributes to the pathogenesis of GC. The viability, migration, and invasion of GC cells were analyzed by CCK-8 and transwell assays. Each condition was repeated three times. The regulation of H3K18la on transcription activity of CNTNl was evaluated by·dua-luciferase reporter assay. Animal experiment was performed using nude mice with six mice in each group, and tumor growth was evaluated. Statistical analysis was performed using GraphPad Prism software with t-test, one-way or two-way ANOVA. We found that PFKM was over-expressed in GC. Downregulated PFKM restrained the viability, migration, invasion, glucose uptake, and lactate production of GC cells. Mechanically, PFKM interacted with CNTN1 and facilitated the enrichment of H3K18la at the CNTN1 promoter region. Overexpression of CNTN1 reversed the inhibitory effects of PFKM knockdown on GC progression. Our research showed that increasing PFKM levels accelerated GC development by regulating CNTN1 expression through mechanisms involving histone lactylation, which could potentially contribute to novel approaches in diagnosing and treating GC.

Human Nervous System-Based Biohybrid Robot-On-A-Chip with Sensing Function for Toxicity Screening.

Biohybrid robots and biohybrid robot-on-a-chip have been developed for drug screening and toxicity screening the complementing animal experiments. A sensing system is needed to evaluate the response to external stimulation, but until now, only the human motor system-based biohybrid robot-on-a-chip with neuromuscular system has been developed without a sensing system. A human nervous system-based biohybrid robot-on-a-chip with eye function as a sensing system in addition to brain/motor neuron/muscle functions is proposed for the first time. Eye assembloid is fabricated by a combination of thalamic organoid covered with Au nanomesh and four retinal organoids. By a combination of eye assembloid, cerebral organoid, motor neuron spheroid, and muscle bundle on polymer substrate, a human nervous system-based biohybrid robot-on-a-chip is made. When blue light is used to cause retinal damage or a hydroxychloroquine (HCQ), a retinal toxic chemical, is applied to the eye assembloid, they caused a decrease in muscle bundle contraction. These results indicated that electrophysiological signals generated by the eye assembloid are transmitted to the muscle bundle through cerebral organoid and motor neuron spheroids, and thus the proposed system can perform the toxicity screening. Human nervous system-based biohybrid robot-on-a-chip can be applied to drug screening of neurodegenerative diseases and toxicity screening for the complement of animal experiments in the future.

Potential of LP as a Biocontrol Agent for Vibriosis in Abalone Farming

Vibrio species are among the primary pathogenic bacteria affecting abalone aquaculture, posing significant threats to farming practices. Current clinical control predominantly relies on antibiotics, which can result in antibiotic residues in both abalone and the surrounding marine environments. Lactobacillus plantarum (LP) has been shown to release bioactive antagonistic substances and exhibits potent inhibitory effects against marine pathogenic bacteria. This study aimed to screen and characterize the probiotic properties of LP strains isolated from rice wine lees to develop a novel biocontrol strategy against Vibriosis in abalone. The methods employed included selective media cultivation, streak plate isolation, and single-colony purification for strain screening, followed by Gram staining, 16S rDNA sequencing, and phylogenetic tree construction using MEGA11 for identification. The resilience, antimicrobial activity, and in vivo antagonistic efficacy of the strains were evaluated through stress tolerance assays, agar diffusion tests, and animal experiments. The results demonstrated the successful isolation and purification of four LP strains (NDMJ-1 to NDMJ-4). Phylogenetic analysis revealed closer genetic relationships between NDMJ-3 and NDMJ-4, while NDMJ-1 and NDMJ-2 were found to be more distantly related. All strains exhibited γ-hemolytic activity, bile salt tolerance (0.3–3.0%), and resistance to both acid (pH 2.5) and alkali (pH 8.5), although they were temperature sensitive (inactivated above 45 °C). The strains showed susceptibility to most of the 20 tested antibiotics, with marked variations in hydrophobicity (1.91–93.15%) and auto-aggregation (13.29–60.63%). In vitro antibacterial assays revealed that cell-free supernatants of the strains significantly inhibited Vibrio parahaemolyticus, V. alginolyticus, and V. natriegens, with NDMJ-4 displaying the strongest inhibitory activity. In vivo experiments confirmed that NDMJ-4 significantly reduced mortality in abalone infected with V. parahaemolyticus. In conclusion, the LP strains isolated from rice wine lees (NDMJ-1 to NDMJ-4) possess robust stress resistance, adhesion capabilities, and broad antibiotic susceptibility. Their metabolites exhibit significant inhibition against abalone-pathogenic Vibrios, particularly NDMJ-4, which demonstrates exceptional potential as a candidate strain for developing eco-friendly biocontrol agents against Vibriosis in abalone aquaculture.

Precancerous pathways to gastric cancer: a review of experimental animal models recapitulating the correa cascade

Gastric cancer remains a significant global health challenge, representing the fifth most commonly diagnosed cancer and the fourth leading cause of cancer-related mortality worldwide. Understanding the pathogenesis of precancerous lesions is crucial for developing effective prevention and treatment strategies. This review provides a comprehensive analysis of animal models for gastric precancerous lesions, categorizing them into Helicobacter infection models, chemical carcinogen or diet-induced models, multifactorial induction models, chemical injury models, and genetically engineered mouse models. We evaluate the advantages and limitations of each model type, with particular focus on their ability to recapitulate the Correa cascade of human gastric carcinogenesis. While Helicobacter felis infection in C57BL/6 mice most closely mirrors the progression from chronic gastritis through metaplasia to dysplasia, these models primarily produce spasmolytic polypeptide-expressing metaplasia rather than true intestinal metaplasia, limiting translational relevance. Chemical carcinogen models reliably produce tumors but often bypass intermediate precancerous stages. Recent advances in genetic engineering, particularly stomach-specific inducible Cre recombinase systems targeting gastric progenitor cells, have yielded models that faithfully reproduce the spectrum of human gastric cancer subtypes with features of metastatic disease. We highlight the importance of standardized histopathological evaluation methodologies and discuss future research directions, including integration of advanced technologies such as single-cell RNA sequencing with existing animal models, development of organoid models, and investigation of interactions among genetic predisposition, Helicobacter infection, and environmental factors. This review provides a valuable reference for researchers investigating gastric precancerous lesions and offers insights for the development of more effective prevention and treatment strategies.

БІОХІМІЧНІ МАРКЕРИ МІНЕРАЛЬНОЇ ЩІЛЬНОСТІ КІСТКОВОЇ ТКАНИНИ У КРОВІ ЕКСПЕРИМЕНТАЛЬНИХ ТВАРИН ІЗ МОДЕЛЬОВАНИМ ОСТЕОПОРОЗОМ

One of the most important components of evidence-based medicine is conducting experimental studies on animals. This allows us to identify the mechanisms of disease development, the effects of drugs, and their toxicity. The aim of our study was to study changes in biochemical markers of bone mineral density in blood in experimental animals with induced osteoporosis when using different osteotropic therapies. In our experimental study, we used 141 rabbits to model glucocorticoid-induced osteoporosis. Osteotropic therapy, developed by us, was carried out starting from the 7th day after the end of osteoporosis modeling. Animals with simulated osteoporotic phenomena were divided into groups depending on the use of pharmacological drugs for reparative osteogenesis: Group I - intact; Group II - without correction of osteoporotic phenomena; Group III - correction of osteoporotic phenomena with the drug "OsteoPro"; Group IV - correction of osteoporosis was performed with the drug "Vitrum Osteomag"; Group V - received a complex of drugs "OsteoPro" + "Vitrum Osteomag".We studied metabolic markers in bone tissue, in particular, the concentration of calcium (Ca) and phosphorus (P), their ratio (Ca/P), the activity of oxyproline, alkaline phosphatase (ALP), acid phosphatase (AP), their ratio, and collagenolytic activity (KA) of blood. The analyzing the obtained data of biochemical markers of bone tissue in experimental animals with modeled osteoporosis, it is possible to state that the use of the combined complex of drugs "OsteoPro + Vitrum Osteomag" allows to significantly improve the biochemical parameters of the blood of animals, which determine the complex cascade of bone tissue remodeling, which was confirmed by the positive dynamics of the parameters studied, both in relation to the data in intact animals and in the intergroup comparison. Thus, a generalized assessment of the Ca/P ratio showed that in intact animals the value of this parameter was 2.02±0.03 and was higher than in animals of other experimental groups: group II – by 8.60% (p≥0.05), group III – by 9.19% (p<0.01), group IV – by 10.99% (p<0.01) and group V – by 13.48%, p<0.01, which indicates resorption processes in bone tissue in osteoporotic phenomena. The generalized bone mineralization index (AlP/AcP) showed that in intact animals of the control group the value of this indicator was 12.79±0.55 points. In experimental animals of group II, where treatment of osteoporotic phenomena was not carried out, the value of AlP/AcP was 17.20% lower compared to the data in animals of group I (p<0.01). In animals of group IV, where treatment of simulated osteoporosis was carried out using the drug "Vitrum Osteomag", the value of AlP/AcP was 4.14% lower than in rabbits of the control group I (p>0.05). The highest value of the mineralization index was studied in experimental animals of group V, where treatment of osteoporotic phenomena was carried out with a combination of drugs "OsteoPro + Vitrum Osteomag" - 19.28±0.39 points. These processes were also confirmed results of studies of the level of collagenolytic activity of blood plasma and oxyproline in animals with modeled osteoporosis. The use of the above-mentioned drugs separately in the treatment of modeled osteoporosis did not cause a convincing improvement in the values ​​of bone remodeling markers.

The efficacy and acceptability of Lactobacillus reuteri for the treatment of depression: A systematic review and meta-analysis.

The efficacy and acceptability of Lactobacillus reuteri for the treatment of depression: A systematic review and meta-analysis.

Association of PM2.5 and its elemental components with plasma metabolome in healthy older adults: Evidence from epidemiological and experimental studies.

Association of PM2.5 and its elemental components with plasma metabolome in healthy older adults: Evidence from epidemiological and experimental studies.

Essential roles of sensory nerve in maintenance of cornea-phenotype in mice.

Essential roles of sensory nerve in maintenance of cornea-phenotype in mice.

Acute effects of peripherally administered nicotine on food intake via the central anorectic peptide nesfatin-1/nucleobindin-2 in adult male rats.

Acute/chronic exposure to nicotine modulates feeding behavior in experimental animals and humans. However, how nicotine modulates food intake remains unclear. This study examined the acute effects of the peripheral administration of nicotine on food intake in adult male rats, focusing on the possible involvement of the anorectic peptide nesfatin-1/nucleobindin-2 (NucB2) in the central nervous system (CNS). Initially, cumulative food intake, but not water intake, was significantly decreased 0.5, 1, and 1.5 h after the intraperitoneal administration of nicotine (0.5 mg/kg) in24h-fasted rats. Subsequently, the double-labeled immunohistochemical study revealed that nesfatin-1/NucB2-immunoreactive (ir) neurons expressed Fos-ir in various nuclei of the hypothalamic and brainstem areas, including the supraoptic nucleus and ventral tegmental areas, 90 min after the intraperitoneal administration of nicotine. Finally, pretreatment with intracerebroventricular administration of antisense RNA against nesfatin-1/NucB2 significantly attenuated the suppression of food intake induced by intraperitoneal nicotine administration. The results indicated that the acute effects of peripherally administered nicotine on the suppression of food intake may be partially involved in nesfatin-1/NucB2-containing neurons in the CNS in male adult rats.

Identification of a translatable animal model for dry eye disease using comparative analysis of tear proteins across species.

This study aimed to assess the similarity of tear proteins between experimental animals and humans to identify the most translational animal model for dry eye disease (DED). Eleven species were selected for a structural and physicochemical comparison of healthy human tear fluid proteins involved in DED. Amino acid sequences were compared using BLAST. Protein primary structure, isoelectric point (pI) and grand average of hydropathicity (GRAVY) were determined using ExPASy and compared with humans. Among non-primate mammals, the cat (69.7%) and pig (68.7%) showed the highest protein sequence similarity to humans. The ruminants and cat showed amino acid content changes for the highest number of proteins (≥3/15). The pig, rabbit, dog and rodents had the closest global pI values to humans, while the cat showed the highest protein number (9/15) with pI values far from humans. GRAVY values for the pig and dog were the closest to humans. Tear-soluble factor study revealed that the pig was the only species with high similarity for all proteins (>60%). Amino acid content was similar for most species compared to humans, except mouse for IL-6 and rodents and pig for IL-8. The pI and GRAVY values varied across species, though the pig and sheep were the only ones with similar pI to humans for four out of five factors. The pig exhibited the highest similarity to humans in tear protein analysis among non-primate mammals, suggesting that the porcine model may be the most translational for DED research.

METHOD FOR DETERMINING THE VIABILITY OF MESENCHYMAL STEM CELLS IN TISSUE-ENGINEERED STRUCTURES DURING REGENERATION – EXPERIMENTAL SUBSTANTIATION OF EFFICIENCY

HighlightsThe method for determining the viability of mesenchymal stem cells in the composition of tissue-engineering constructs is proposed here. This method can become an effective tool for regenerative medicine and tissue engineering for assessing the mechanisms of the regenerative process when using tissue-engineered constructs and predicting their effectiveness in an in vitro system. AbstractAim. Еxperimental substantiation of the efficiency of the method for determining the viability of mesenchymal stem cells in tissue-engineered constructs during regenerationMethods. The experimental study was conducted on 12 animals (rats). A full-layer scalped wound with a diameter of 3 cm was formed in the animals. A tissue-engineering construct based on human plasma cryoprecipitate was implanted into the wound, with the composition of which a suspension of rat mesenchymal stem cells (MSCs) pre-stained with the DiOC14(3) tracer was introduced. At the control periods (3, 7–8, 14 and 21 days), the animals were withdrawn from the experiment. During the operation, the wound defect area was cut out for ex-tempore examination using wide-field fluorescence microscopy with visualization of samples with 40×, 100×, 200× magnification in the GFP fluorescence channel.Results. When examining the material obtained from animals on day 3, the external characteristics of the overwhelming majority of identified objects with characteristic fluorescent glow corresponded to the morphological characteristics of MSCs – spindle-shaped cells with characteristic processes. In the material obtained from experimental animals on days 7–8, a large number of cells stained with a tracer fluorescent membrane dye were also detected, which had a green fluorescent glow and characteristic morphology. In the biomaterial obtained on day 14 of the experiment, the number of objects with characteristic MSC morphology stained with the DiOC14(3) tracer was significantly less than on days 3 and 7–8 of the experiment. On day 21, a signal specific for the DiOC14(3) tracer was also detected, but it was not possible to visualize even single cells with the morphology characteristic of MSCs; the signal was probably due to fragments of membranes of eliminated MSCs labeled with a fluorescent dye.Conclusion. It was shown that MSCs in tissue-engineering constructs implanted on the animal wound retain viability and characteristic morphology for at least 14 days. It is important that the proposed method allows intravital visualization and photofixation of viable MSCs with characteristic morphology, and thus avoiding distortions that may be associated with fixation of biological material. The proposed method can become an effective tool for regenerative medicine and tissue engineering in relation to assessing the mechanisms of the regenerative process using TECs and predicting their effectiveness in the in vitro system.

In vivo evaluation of three isoxazolines against Cx. tritaeniorhynchus (Diptera: Culicidae): A novel approach to control Japanese encephalitis vector.

In vivo evaluation of three isoxazolines against Cx. tritaeniorhynchus (Diptera: Culicidae): A novel approach to control Japanese encephalitis vector.

Chemical Characterization, Antidiabetic and Antioxidant Potential of Allium cepa Peels Silver Nanoparticles Against Alloxan Induced Diabetes in Rabbits.

Diabetes is one of the major causes of death worldwide. Onion peels have antioxidant and antidiabetic activities due to the presence of flavonoids and phenolic compounds. In current research, the antidiabetic potential of a herbal-based silver nanodrug synthesized using aqueous extract of Allium cepa peels was studied against Alloxan-induced diabetes in rabbits. The green synthesized reduced silver nanoparticles (AgNPs) and aqueous extract were characterized using X-ray diffraction analysis (XRD) and liquid chromatography mass spectrometry (LCMS-MS) analysis. After the acclimation period, the rabbits were divided into nine groups of 5 rabbits each (n = 5). Experimental animals were injected with a single dose of Alloxan monohydrate (160 mg/mL per kg BW) and then orally treated with aqueous extract of A. cepa peels and AgNPs at three dose concentrations (100, 200, 300 mg/mL per kg BW) for 20 days. Serum levels of blood sugar, renal function tests, lipid profile, and oxidative stress markers were determined photometrically. Results from mass spectral analysis revealed that quercetin and vanillic acid are the major phenolic compounds. XRD analysis confirmed the stabilization and shape of green synthesized AgNPs. Results from invivo studies indicated that treatment with aqueous extract of A. cepa peels and AgNPs attenuated the Alloxan-induced diabetes in a dose-dependent manner, as demonstrated by a decrease in blood sugar, triglycerides, urea, creatinine, and cholesterol to 100 ± 0.8, 57.66 ± 0.8, 38 ± 0.8, 1.11 ± 0.08, 40 ± 0.00 mg/dL, respectively, in rabbits administered with 300 mg/mL per kg BW AgNPs. The levels of oxidative stress markers (CAT, SOD, and GPx) in blood, that is, 3.6 ± 0.08, 6.0 ± 0.08, 918 ± 2.0 IU/mL, respectively, were also increased in rabbits administered with 300 mg/mL per kg BW AgNPs. Our results conclude that synthesized AgNPs using aqueous extract of A. cepa peels have a protective effect against Alloxan-induced diabetes.

Biological Estimation of Phenolic-flavonoid Contents for In vitro Antioxidant Activity and Antidiabetic Activities of Selaginella plana in Experimental Models

Introduction: The study aimed to assess the effectiveness of Selaginella plana in treating diabetes. The plant components were assessed using ethanol as a solvent. To determine whether the plant extracts included any secondary metabolites, a phytochemical screening was performed. Methods: The quantities of total phenolic and total flavonoid were determined using plant extracts in ethanolic, chloroform, petroleum, and water solutions. Experimental animals were used to evaluate the antioxidant properties of plant extracts. Thus, the study was further processed to evaluate antidiabetic activity using ethanolic and aqueous extracts. Glucose levels were measured using an oral glucose tolerance test (OGTT). Results: The percentage yield of ethanol, chloroform, petroleum ether, and aqueous extract were 19.22 g, 11.01 g, 6.44 g, and 15.76 g, respectively. The ethanolic and aqueous extracts showed the presence of most of the phytoconstituents like alkaloids, flavonoids, carbohydrates, tannins, phytosterols, glycosides, proteins, and gum. TPC values were high for ethanolic extract of the fern S. plana, which was 54.34 mg GAE/g for ethanolic extract. However, the TFC value was 264.51 mg QE/g for ethanolic extract. Normalisation of insulin levels and restoration of blood glucose levels were both demonstrated by the plant extract. Conclusion: The major findings of the current study demonstrated that ethanolic extract of S. plana was effective against STZ-induced diabetes. S. plana may be an alternative drug in treating diabetes after clinical trials.

Integrating metabolomics and network pharmacology analysis provides new insights into the lipid metabolism disorder of Abrus mollis.

Integrating metabolomics and network pharmacology analysis provides new insights into the lipid metabolism disorder of Abrus mollis.

MCPA-Na exposure in aquatic systems: disruption of pathways and increased susceptibility to infection in fish.

MCPA-Na exposure in aquatic systems: disruption of pathways and increased susceptibility to infection in fish.

Melanogenic activity of fish scale-derived peptides FTGML from grass carp:An integrated study of molecular docking, network pharmacology and animal experiments

Melanogenic activity of fish scale-derived peptides FTGML from grass carp:An integrated study of molecular docking, network pharmacology and animal experiments

Recombinant adenovirus expressing pdhβ-pdhD fusion protein produces robust immune responses and partial protection against Mycoplasma Synoviae challenge in chickens.

Recombinant adenovirus expressing pdhβ-pdhD fusion protein produces robust immune responses and partial protection against Mycoplasma Synoviae challenge in chickens.

SP1-Mediated Upregulation of KIF4A Promotes Prostate Cancer Progression via Targeting TWIST1.

Previous studies have found the highly expressed kinesin family member 4A (KIF4A) in various tumors, including prostate cancer. However, the role of KIF4A and its potential mechanisms in the development of prostate cancer remains unclear. Bioinformatics analysis was used to analyze the levels of KIF4A and TWIST-related protein 1 (TWIST1) based on the data from TCGA, HPA, and ENCORI databases. RT-qPCR was performed to measure the mRNA expression of KIF4A and TWIST1. The protein expression of KIF4A and specificity protein 1 (SP1) was examined through western blot analysis. CCK-8, EdU, and colony formation assay were carried out to evaluate the proliferative ability. The invasive and migrated cells were determined using the Transwell invasion assay and the wound-healing assay, respectively. EMT progress was evaluated by detecting E-cadherin and N-cadherin expression using western blot analysis. The relationship between SP1 and KIF4A was verified using ChIP and dual-luciferase reporter assay. The interaction between KIF4A and TWIST1 was confirmed through Co-IP. Animal experiments and IHC were carried out to explore the effects of KIF4A on prostate cancer mice in vivo. KIF4A was increased in prostate cancer tissues and cells, and KIF4A knockdown suppressed the proliferative, invaded, and migrated abilities, as well as EMT in DU145 and PC3 cells. The inhibition effects of SP1 knockdown on cell proliferation, invasion, migration, and EMT were harbored by overexpressing KIF4A. TWIST1 was elevated in prostate cancer and positively regulated by KIF4A. TWIST1 upregulation reversed the repressive impacts of KIF4A downregulation on prostate cancer progression. In addition, in vivo experiments and IHC assay indicated that KIF4A depletion inhibited tumor growth and EMT in mice. Our findings suggested that SP1 activated KIF4A to regulate TWIST1 expression, thereby promoting the development of prostate cancer.

Anti-inflammatory effects of Rehmannia glutinosa polysaccharide on LPS-induced acute liver injury in mice and related underlying mechanisms.

Anti-inflammatory effects of Rehmannia glutinosa polysaccharide on LPS-induced acute liver injury in mice and related underlying mechanisms.