Prophylactic Pattern Scanning Laser Retinal Photocoagulation for Diabetic Retinopathy in Spontaneously Diabetic Torii Fatty Rats: Preliminary Experimental Results

Research Background and Purpose: The number of diabetic patients is rapidly increasing, making it crucial to find methods to prevent diabetic retinopathy (DR), a leading cause of blindness. We investigated the effects of prophylactic pattern scanning laser retinal photocoagulation on DR development in Spontaneously Diabetic Torii (SDT) fatty rats as a new prevention approach. Methods: Photocoagulation was applied to the right eyes of 8-week-old Spontaneously Diabetic Torii (SDT) fatty rats, with the left eyes serving as untreated controls. Electroretinography at 9 and 39 weeks of age and pathological examinations, including immunohistochemistry for vascular endothelial growth factor and glial fibrillary acidic protein at 24 and 40 weeks of age, were performed on both eyes. Results: There were no significant differences in amplitude and prolongation of the OP waves between the right and left eyes in SDT fatty rats at 39 weeks of age. Similarly, no significant differences in pathology and immunohistochemistry were observed between the right and left eyes in SDT fatty rats at 24 and 40 weeks of age. Conclusion: Prophylactic pattern scanning retinal laser photocoagulation did not affect the development of diabetic retinopathy in SDT fatty rats.

Effect of Mushroom Extract Hericium erinaceus on Spatial Memory and Morphology of Neurons in the CA1 and CA3 Regions of the Hippocampus in Ovariectomized Rats

Various studies indicate that low levels of estradiol negatively impact cognitive abilities. Extracts from the fungus Hericium erinaceus (HE) contain bioactive components that promote the proper functioning of the nervous system and potential effects on protection against neurodegenerative diseases, including dementia and motor dysfunctions. The objective was to evaluate the effects of the administration of the HE mushroom extract on visuospatial memory and morphology of neurons in the CA1 and CA3 regions of the hippocampus in ovariectomized rats. 40 young Wistar rats weighing 90 ± 10 g BW were used, which were distributed into four groups of 10 animals;Control Group, non-ovariectomized and untreated rats;Group E2, ovariectomized rats treated with estradiol (2 μg/kg/body weight);Group HE, ovariectomized rats treated with the extract of the fungus Hericium erinaceus (0.5 mg/kg body weight) and Group Ovx/ST, ovariectomized rats, without treatment. The animals were tested in the Barnes and Open Field maze, then they were sacrificed, and their brains were obtained to perform a histological analysis of neuronal morphology in the CA1 and CA3 areas of the hippocampus. The most outstanding results showed that the Ovx/ST group recorded the longest time to arrive at the escape box and stay in the Barnes maze. A correlation was observed between neuronal damage and function;in the groups that did not present satisfactory performance in the maze tests, morphological alterations were identified such as the presence of some neuronal somata with degeneration characteristics such as pyknosis, nuclear basophilia and shrinkage of the cells. Its soma, as well as a decrease in the nuclear area of CA1 and CA3 neurons. It is concluded that the fungus Hericium erinaceus exerted a neuroprotective effect on the neuronal bodies of the hippocampus, associated with better performance in the visuospatial recognition memory test.

Long-Term Impact of Acute Retinoic Acid Supplementation at the Young Age on Testicular Architecture of Wistar Albino Rats

Introduction: Inappropriate and excess vitamin supplementation, particularly for vitamin A, is increasingly recognized as a public health problem in developed countries. On the other hand, blind supplementation of vitamin A, for children in developing countries is a subject of controversy in the literature. The crucial role of vitamin A in the process of spermatogenesis in adult rodents is well established, but only a few publications are consecrated to the long-term effect of vitamin A intake at a young age on testicular development and differentiation. Objectives: Our study aimed to evaluate the long-term effects of acute supplementation at an early age, in the post-natal period, on spermatogenesis and testicular trophicity at adult age. Material and Methods: Young Wistar Albinos rats of 22 days received an acute high dose of supplementation of vitamin A (retinyl palmitate). The control group, group 1, received only extra virgin olive oil, Group 2 a dose of 7000 IU/kg of retinyl palmitate, group 3, 14,000 IU/kg, and Group 4 a dose of 28,000 IU/kg. At 10 weeks of age, the testes’ testosterone levels were measured by ELISA. For histological assessment, sections were stained with Hematoxylin eosin, and the Johnsen score was used to evaluate spermatogenesis in the seminiferous tubules. Results: The average testicular weights of rats were significantly lower in group 4 (p < 0.05), and so was the testosterone level in the testis compared to the control group (p .01). Most of the seminiferous tubules were concerned by an arrest of spermatogenesis and the Johnsen score was decreased with a mean score of 5.96 ± 1.60 (p .001) in that Group. In Group 3, Johnsen’s score was significantly better than the one obtained with the control. Conclusion: We observed a negative effect in the long term with a high acute dose of supplementation of retinyl palmitate at a young age, on testicular development and differentiation. Despite a return to normal diet after that supplementation, during childhood, impaired spermatogenesis was identified at the adult age with an arrest of spermatogenesis. The reversibility of that lack of differentiation by a return to a normal diet is questionable and would need more investigation.

Underlying anti-hypertensive mechanism of the Mizuhopecten yessoensis derived peptide NCW in spontaneously hypertensive rats via widely targeted kidney metabolomics

The angiotensin-converting enzyme(ACE)inhibitory peptide NCW derived from Mizuhopecten yessoensis has been demonstrated to have significant in vivo anti-hypertensive effects,however,its anti-hypertensive mechanism is still not fully clarified.This study established a UPLC-Q-TRAP-MS/MS-based widely targeted kidney metabolomics approach to explore the changes of kidney metabolic profiles and to clarify the antihypertensive mechanism of peptide NCW in spontaneously hypertensive rats(SHRs).Multivariate statistical analysis indicated that the kidney metabolic profiles were clearly separated between the SHR-NCW and SHRUntreated groups.A total of 85 metabolites were differentially regulated,and 16 metabolites were identified as potential kidney biomarkers,e.g.,3-hydroxybutyrate,malonic acid,deoxycytidine,and L-aspartic acid.The peptide NCW might regulate kidney metabolic disorder of SHRs to alleviate hypertension by suppressing inflammation and improving nitric oxide production under the regulation of linoleic acid metabolism,folate related pathways,synthesis and degradation of ketone bodies,pyrimidine metabolism,β-alanine metabolism,and retinal metabolism.

Identifying the stability of housekeeping genes to be used for the quantitative real-time PCR normalization in retinal tissue of streptozotocin-induced diabetic rats

AIM:To investigate the stability of the seven housekeeping genes:beta-actin(ActB),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),18s ribosomal unit 5(18s),cyclophilin A(CycA),hypoxanthine-guanine phosphoribosyl transferase(HPRT),ribosomal protein large P0(36B4)and terminal uridylyl transferase 1(U6)in the diabetic retinal tissue of rat model.METHODS:The expression of these seven genes in rat retinal tissues was determined using real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)in two groups;normal control rats and streptozotocininduced diabetic rats.The stability analysis of gene expression was investigated using geNorm,NormFinder,BestKeeper,and comparative delta-Ct(ΔCt)algorithms.RESULTS:The 36B4 gene was stably expressed in the retinal tissues of normal control animals;however,it was less stable in diabetic retinas.The 18s gene was expressed consistently in both normal control and diabetic rats’retinal tissue.That this gene was the best reference for data normalisation in RT-qPCR studies that used the retinal tissue of streptozotocin-induced diabetic rats.Furthermore,there was no ideal gene stably expressed for use in all experimental settings.CONCLUSION:Identifying relevant genes is a need for achieving RT-qPCR validity and reliability and must be appropriately achieved based on a specific experimental setting.

Pharmacokinetics of Gelsemium elegans in female rats

Background:Gelsemium elegans Benth(G.elegans)is a poisonous perennial evergreen vine plant that has been applied in livestock production and veterinary clinical practice.Early studies found that the toxicity of G.elegans showed significant gender differences in rats,but the underlying reasons for this difference are still not well understood.Methods:In order to explore whether the gender differences in the toxicity of G.elegans are related to pharmacokinetic differences,based on the previous pharmacokinetic study of multiple components of G.elegans in male rats,this study used HPLC-MS/MS method established in the laboratory to conduct a pharmacokinetic study of multiple alkaloids in the plasma of female rats after a single gavage administration of G.elegans(dose of 0.1 g/kg).Results:Through detection,17 alkaloid components in the plasma of female rats were identified,and the pharmacokinetic parameters of 11 of these alkaloids were calculated.We find that in female rats.The T_(max)values were generally less than 0.5 h,and the T_(1/2)values exceeded 3 h,with the longest reaching up to 32.80 h half elimination time.Additionally,the C_(max)and AUC results indicated that female rats had generally higher absorption and exposure levels for most alkaloids.Conclusion:These results suggest that the reason for the differences in the toxicology of G.elegans may be related to the absorption and exposure of gelsemidine-type alkaloids in animals.

Study on the Therapeutic Effects and Mechanisms of Human Mesenchymal Stem Cell-Derived Exosomes Carrying NGF Gene in Treating Ischemic Stroke in Rats

Objective:To investigate the therapeutic effects and mechanisms of human mesenchymal stem cell-derived exosomes(hMSCs-Exo)carrying the NGF gene in treating ischemic stroke in rats,aiming to provide new insights and treatment methods for ischemic stroke therapy.Methods:After successful construction of the cerebral ischemia model in 40 male SPF-grade SD rats aged 6-8 weeks,the model rats were randomly divided into 4 groups:Sham group,PBS group,hMSCs-Exo group,and NGF-hMSCs-Exo group,with 10 rats in each group.The rat MCAO model was prepared using the classic filament method,and NGF-hMSCs-Exo were injected via the tail vein into the MCAO model rats.The expression of the NGF gene in brain ischemic tissues,neuronal regeneration,and rat neurological function recovery were observed using TTC staining,memory function evaluation,Western blot,qRT-PCR,and other methods.Results:Compared with the Sham group,neurological deficits were significant in the PBS group(P<0.01).Compared with the PBS group,neurological scores improved in the hMSCs-Exo group and NGF-hMSCs-Exo group(P<0.05).Compared with the hMSCs-Exo group,the improvement in neurological deficits was more significant in the NGF-hMSCs-Exo group(P<0.05).The infarct area after NGF-hMSCs-Exo intervention was significantly reduced(P<0.05)compared with the Sham group.Compared with the PBS group,relative expression levels of NGF mRNA and protein decreased,while Caspase-3 mRNA and protein expression significantly increased in the PBS group(P<0.01).Compared with the PBS group and hMSCs-Exo group,there were differences in NGF and Caspase-3 mRNA and protein expression in the NGF-hMSCs-Exo group rat brain tissues(P<0.05).Conclusion:Treatment with human mesenchymal stem cell-derived exosomes carrying the NGF gene improves cognitive function and exerts protective effects on SD rats while inhibiting apoptotic levels in cells.

Finding biomarkers of experience in animals

At a time when there is a growing public interest in animal welfare,it is critical to have objective means to assess the way that an animal experiences a situation.Objectivity is critical to ensure appropriate animal welfare outcomes.Existing behavioural,physiological,and neurobiological indicators that are used to assess animal welfare can verify the absence of extremely negative outcomes.But welfare is more than an absence of negative outcomes and an appropriate indicator should reflect the full spectrum of experience of an animal,from negative to positive.In this review,we draw from the knowledge of human biomedical science to propose a list of candidate biological markers(biomarkers)that should reflect the experiential state of non-human animals.The proposed biomarkers can be classified on their main function as endocrine,oxidative stress,non-coding molecular,and thermobiological markers.We also discuss practical challenges that must be addressed before any of these biomarkers can become useful to assess the experience of an animal in real-life.

Anti-Obesity Effects of Dietary d-Allulose and Medium-Chain Triglycerides in High-Fat Diet-Fed Rats

d-Allulose, a rare sugar, exerts anti-obesity effects by inhibiting hepatic lipogenesis and promoting energy expenditure. Medium-chain triglycerides (MCTs) consist of three medium-chain fatty acids connected by glycerol. MCTs have been extensively investigated for their ability to reduce body fat accumulation. We previously investigated the anti-obesity effects of a combination of dietary d-allulose and MCT (5% - 13%) in rats;however, we could not confirm the anti-obesity effects of MCT or observed synergetic effects between d-allulose and MCT on body fat loss. We speculated that our previous studies were influenced by the excessive amount of MCT in the diets. Therefore, in this study, we aimed to investigate the anti-obesity effects of the simultaneous intake of d-allulose and MCT in rats fed an obesity-inducing high-fat diet with a low amount of MCTs (2%). Thirty-two male Wistar rats (3-week-old) were randomly divided into four groups: control, d-allulose, MCT, and d-allulose + MCT groups. Rats in each group were fed ad libitum on a control (no d-Allulose or MCT), 5% d-allulose, 2% MCT, or 5% d-allulose + 2% MCT diets for 16 weeks. Abdominal adipose tissue weights were significantly lower in the d-allulose diet group than in the control group, whereas no differences were observed between results of the MCT-supplemented groups. The total body fat mass was significantly lower in the d-allulose and MCT diet groups than in the control group, but no differences were observed between the MCT-supplemented groups. These results suggested that anti-obesity effects of dietary d-allulose were observed, and the effects of dietary MCTs were weaker than those of d-allulose. Moreover, we confirmed the interaction between dietary d-allulose and MCT on indicators of obesity. Interestingly, their effects were not synergistic, as MCT supplementation offset the anti-obesity effects of dietary d-allulose. However, the specific mechanisms underlying those effects remain unknown, warranting further investigation.

Probiotic microorganisms affect the reproductive and nervous systems of male rats treated with acrylamide

Objective:To evaluate the protective effects of probiotic microorganisms on the reproductive and nervous systems of male rats treated with acrylamide.Methods:Thirty-two rats were randomly divided into 4 groups and received normal saline through gavage(control),acrylamide 20 mg/kg body weight,acrylamide plus probiotic microorganisms(Lactobacillus acidophilus,Lactobacillus casei,Lactobacillus bulgaricus,Lactobacillus rhamnosus,Bifidobacterium breve,Bifidobacterium infantis,Streptococcus thermophilus and fructooligosaccharides,all mixed in sachets)20 or 200 mg/kg body weight,respectively.After 30 days,the testis,prostate,seminal vesicle and cerebellum were removed,fixed and stained with hematoxylin-eosin(H&E).The Johnsen score was used to classify spermatogenesis.Cavalieri's principle method was used to evaluate the total volume(in mm3)of the testes.The number of each intratubular cell type as well as intertubular Leydig cells in whole samples was measured using the physical dissector counting techniques.Stereological analysis and the grids were used to determine the volume of cerebellar layers as well as the Purkinje cell number.Results:The testis weight decreased significantly in the acrylamide-treated group compared to the other groups(P<0.001).The number of spermatogonia,spermatocytes,spermatids and Leydig cells in the acrylamide-treated group were significantly less compared to the control group(P<0.05),while they were increased significantly in the acrylamide+200 mg/kg probiotic group(P<0.05;P<0.01).The mean Johnsen score in the acrylamide-treated group was lower than in the control group(P<0.001).Acrylamide-induced changes including congestion,vacuolization in the secretory epithelial cells,and epithelial rupture were observed in the prostate and seminal vesicle.The volumes of cerebellar layers were decreased in the acrylamide group compared to the control group while recovered in both probiotic treated groups.Conclusions:Probiotic microorganisms alleviate acrylamide-induced toxicities against the reproductive and cerebellar tissues in rats.

Effects of heat stress and long photoperiod on the prostate of rats

Objective:To examine light and heat effects on the morphological,histological,and micrometric structure of the prostate of rats.Methods:Thirty adult male rats were divided into three groups.The control group was kept under 20℃-22℃ and an artificial 12 h/12 h day/night cycle;the temperature group was under normal light and at(42±1)℃ heat for 4 to 5 h daily,and the light group was exposed to 8 h/16 h day/night cycle with 20℃-22℃.Rats were weighed five times(at the beginning of the study and every seven days).Five milliliters(mL)of their peripheral blood were taken.The tissue staining was performed using the hematoxylin-eosin(H&E)stain and periodic acid-Schiff(PAS).In the following,tissue and cellular reactions to the PAS were examined.Results:Folds were located entirely on the surface of the anterior lobe and periphery of the other lobes.The secretory units in the anterior lobe were more than the lateral lobe.A strong reaction of the secretory cells to the PAS was observed.Testosterone serum levels of the light group also significantly increased compared to the control group(P<0.05).The most histometric changes of the lobes were established in the lateral lobes.Heat stress resulted in a significant decrease in testosterone levels and transformed prostate tissue.The epithelium and parenchyma to scaffold ratio in the temperature group decreased.Conclusions:Maximum and minimum changes in the ventral lobe happened under the ascent of temperature and light,respectively.The ventral lobe in the study of prostatic hyperplasia should be more considered.

Development of an experimental method for well-controlled blast induced traumatic limb fracture in rats

Heterotopic ossification(HO)is a consequence of traumatic bone and tissue damage,which occurs in 65%of military casualties with blast-associated amputations.However,the mechanisms behind blast-induced HO remain unclear.Animal models are used to study blast-induced HO,but developing such models is challenging,particularly in how to use a pure blast wave(primary blast)to induce limb fracture that then requires an amputation.Several studies,including our recent study,have developed platforms to induce limb fractures in rats with blast loading or a mixture of blast and impact loading.However,these models are limited by the survivability of the animal and repeatability of the model.In this study,we developed an improved platform,aiming to improve the animal's survivability and injury repeatability as well as focusing on primary blast only.The platform exposed only one limb of the rat to a blast wave while providing proper protection to the rest of the rat's body.We obtained very consistent fracture outcome in the tibia(location and pattern)in cadaveric rats with a large range of size and weight.Importantly,the rats did not obviously move during the test,where movement is a potential cause of uncontrolled injury.We further conducted parametric studies by varying the features of the design of the platform.These factors,such as how the limb is fixed and how the cavity through which the limb is placed is sealed,significantly affect the resulting injury.This platform and test setups enable well-controlled limb fracture induced directly by pure blast wave,which is the fundamental step towards a complete in vivo animal model for blast-induced HO induced by primary blast alone,excluding secondary and tertiary blast injury.In addition,the platform design and the findings presented here,particularly regarding the proper protection of the animal,have implications for future studies investigating localized blast injuries,such as blast induced brain and lung injuries.

The water-soluble TF3 component from Eupolyphaga sinensis Walker promotes tibial fracture healing in rats by promoting osteoblast proliferation and angiogenesis

Objective:To determine the active components of Eupolyphaga sinensis Walker(Tu Bie Chong)and explore the mechanisms underlying its fracture-healing ability.Methods: A modified Einhorn method was used to develop a rat tibial fracture model.Progression of bone healing was assessed using radiological methods.Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site.Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration.The Transwell assay was used to explore the invasion capacity of the cells.Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells(HUVECs).qRT-PCR was used to evaluate the changes in gene transcription levels.Results: Tu Bie Chong fraction 3(TF3)significantly shortened the fracture healing time in model rats.X-ray results showed that on day 14,fracture healing in the TF3 treatment group was significantly better than that in the control group(P=.0086).Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group.In vitro,TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs(all P<.01).Transwell assays showed that TF3 promoted the migration of HUVECs,but inhibited the migration of MC3T3-E1 cells.Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules.Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA,SPOCD1,NGF,and NGFR in HUVECs.In MC3T3-E1 cells,the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment.Conclusion: TF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.

Dry environment on the expression of lacrimal gland S100A9,Anxa1,and Clu in rats via proteomics

●AIM:To investigate the underlying mechanism of dry environment(autumn dryness)affecting the lacrimal glands in rats.●METHODS:Twenty Sprague-Dawley rats were randomly divided into two groups.The rats were fed in specific pathogen free environment as the control group(n=10),and the rats fed in dry environment as the dryness group(n=10).After 24d,lacrimal glands were collected from the rats.The tissues morphology was observed by hematoxylineosin(HE)staining.Tandem mass tags(TMT)quantitative proteomics analysis technology was used to screen the differential expressed proteins of lacrimal glands between the two groups,then bioinformatics analysis was performed.Further,the immunohistochemical(IHC)method was used to verify the target proteins.●RESULTS:In dryness group,the lacrimal glands lobule atrophied,the glandular cavities enlarged,the sparse nuclear distribution and scattered inflammatory infiltration between the acinus were observed.The proteomics exhibited that a total of 195 up-regulated and 236 downregulated differential expressed proteins screened from the lacrimal glands of rats.It was indicated that the biological processes(BP)of differential expressed proteins mainly included cell processes and single BP.The cellular compositions of differential expressed proteins mainly located in cells,organelles.The molecular functions of differential expressed proteins mainly included binding,catalytic activity.Moreover,the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis showed that the differential expressed proteins mainly involved lysosome,complement and coagulation cascade,and ribosome pathway.The IHC result verified that the up-regulated expression proteins of Protein S100A9(S100A9),Annexin A1(Anxa1),and Clusterin(Clu)in lacrimal glands of rats in dryness group were higher than control group.●CONCLUSION:The up-regulated expression proteins of S100A9,Anxa1,and Clu may be the potential mechanisms of dry eye symptoms caused by dry environment.This study provides clues of dry environments causing eye-related diseases for further studies.

Xuebijing improves intestinal microcirculation dysfunction in septic rats by regulating the VEGF-A/PI3K/Akt signaling pathway

BACKGROUND:This study aims to explore whether Xuebijing(XBJ) can improve intestinal microcirculation dysfunction in sepsis and its mechanism.METHODS:A rat model of sepsis was established by cecal ligation and puncture(CLP).A total of 30 male SD rats were divided into four groups:sham group,CLP group,XBJ + axitinib group,and XBJ group.XBJ was intraperitoneally injected 2 h before CLP.Hemodynamic data(blood pressure and heart rate) were recorded.The intestinal microcirculation data of the rats were analyzed via microcirculation imaging.Enzyme-linked immunosorbent assay(ELISA) kits were used to detect the serum levels of interleukin-6(IL-6),C-reactive protein(CRP),and tumor necrosis factor-α(TNF-α) in the rats.Histological analysis and transmission electron microscopy were used to analyze the injury of small intestinal microvascular endothelial cells and small intestinal mucosa in rats.The expression of vascular endothelial growth factor A(VEGF-A),phosphoinositide 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(Akt),and phosphorylated Akt(p-Akt) in the small intestine was analyzed via Western blotting.RESULTS:XBJ improved intestinal microcirculation dysfunction in septic rats,alleviated the injury of small intestinal microvascular endothelial cells and small intestinal mucosa,and reduced the systemic inflammatory response.Moreover,XBJ upregulated the expression of VEGF-A,p-PI3K/total PI3K,and p-Akt/total Akt in the rat small intestine.CONCLUSION:XBJ may improve intestinal microcirculation dysfunction in septic rats possibly through the VEGF-A/PI3K/Akt signaling pathway.

Oral administration of egg white ovotransferrin prevents osteoporosis in ovariectomized rats

Ovotransferrin,an iron-binding glycoprotein,accounting for approximately 12%of egg white protein,is a member of transferrin fam ily.Our previous studies showed that ovotransferrin stimulates the proliferation and differentiation of osteoblasts,while inhibits osteoclastogenesis and resorption activity.The work aims to study the efficacy of orally administered ovotransferrin on the prevention of osteoporosis using ovariectomized(OVX)Sprague-Dawley rats.Oral administration of ovotransferrin showed no negative effect on body weight,food intake and organ weight.After 12-week treatment,feeding ovotransferrin at a dose of 1%(1 g ovotransferrin/100 g diet)prevented OVX-induced bone loss and maintained relatively high bone mineral density and integrated bone microarchitecture.The serum concentration of biomarkers indicating bone formation was increased in ovotransferrin administration groups,while the bone resorption biomarkers were decreased.Ovotransferrin feeding also decreased the production of serum cytokine TNF-αand IL-6,which are two stimulators for osteoclast differentiation.In addition to its direct regulatory role on bone turnover,ovotransferrin supplementation might benefit osteoporosis prevention by inhibiting adipogenesis,and regulating immune response.Our results suggested the potential application of ovotransferrin as a functional food ingredient on the prevention of osteoporosis.

Influence of different magnetic forces on the effect of colonic anastomosis in rats

BACKGROUND Despite much work having been conducted on magnetic compression anastomo-sis(MCA)in the digestive tract,there are no reports on the influence of magnetic force on the anastomosis.AIM To investigate the effect of different magnetic force magnets on the MCA of the digestive tract.METHODS Two groups of magnets of the same sizes but different magnetic forces were designed and produced.A total of 24 Sprague-Dawley rats were randomly assigned into two groups(powerful magnet group and common magnet group),with 12 rats in each group.Two types of magnets were used to complete the colonic side-to-side anastomosis of the rats.The operation time and magnet discharge time were recorded.The anastomotic specimens were obtained 4 wk after the operation and then the burst pressure and diameter of the anastomosis were measured,and the anastomosis was observed via the naked eye and subjected to histological examination.RESULTS The magnetic forces of the powerful and common magnet groups at zero distance were 8.26 N and 4.10 N,respectively.The colonic side-to-side anastomosis was completed in all 24 rats,and the operation success rate and postoperative survival rate were 100%.No significant difference was noted in the operation time between the two groups.The magnet discharge time of the powerful magnet group was slightly longer than that of the common magnet group,but the difference was not statistically significant(P=0.513).Furthermore,there was no statistical difference in the burst pressure(P=0.266)or diameter of magnetic anastomosis(P=0.095)between the two groups.The gross specimens of the two groups showed good anastomotic healing,and histological observation indicated good mucosal continuity without differences on healing.CONCLUSION In the rat colonic side-to-side MCA model,both the powerful magnet with 8.26 N and the common magnet with 4.10 N showed no significant impact on the anastomosis establishment process or its effect.

Protective Effect of Naringenin on Acute Myocardial Ischemia-reperfusion Injury in Rats

[Objectives]To investigate the protective mechanism of naringenin on acute myocardial ischemia-reperfusion injury(AMI-RI)in Sprague-Dawley(SD)rats.[Methods]A total of 32 SD rats with AMI-RI model construction were randomly divided into AMI-RI model control group and citrus pigment A/B/C groups(n=8).The naringenin A,B,and C groups were administrated 20,40 and 80 mg/(kg•d)for 10 d.The AMI group served as the negative control and was not treated.At the conclusion of the treatment regimen,a sample of intraventricular blood was collected for the purpose of measuring lactate dehydrogenase(LDH),glutathione peroxidase(GLH-PX),nitric oxide(NO),and superoxide dismutase(SOD)levels.Additionally,myocardial tissue was identified within the ischemic region.The content of malondialdehyde(MDA)was determined by inducing nitric oxide synthase(iNOS)and endodermal nitric oxide synthase(eNOS)positive cells in the left anterior descending coronary artery.[Results]Following citrus treatment,the contents of GLH-PX and SOD in ventricular blood of the citrus B group were found to be significantly elevated,while the contents of NO and LDH in myocardial MDA and ventricle were observed to be significantly reduced.The number of eNOS-positive cells was significantly increased,while the number of iNOS-positive cells was significantly decreased.The difference was statistically significant when compared with the AMI-RI group(P<0.05).The changes observed in the above indicators in the citrus C group were more pronounced than those observed in the citrus B group.The difference between the citrus C and the B group was statistically significant(P<0.05),indicating that this effect is concentration dependent.[Conclusions]In addition to its ability to inhibit myocardial lipid peroxidation during AMI-RI by increasing SOD activity,naringenin may also affect the synthesis and release of NO by regulating eNOS and iNOS,thereby achieving protection against AMI-RI.One effect is enhanced as the dose of the drug increases.

N-acetylserotonin alleviates retinal ischemia-reperfusion injury via HMGB1/RAGE/NF-κB pathway in rats

AIM:To observe the effects of N-acetylserotonin(NAS)administration on retinal ischemia-reperfusion(RIR)injury in rats and explore the underlying mechanisms involving the high mobility group box 1(HMGB1)/receptor for advanced glycation end-products(RAGE)/nuclear factor-kappa B(NF-κB)signaling pathway.METHODS:A rat model of RIR was developed by increasing the pressure of the anterior chamber of the eye.Eighty male Sprague Dawley were randomly divided into five groups:sham group(n=8),RIR group(n=28),RIR+NAS group(n=28),RIR+FPS-ZM1 group(n=8)and RIR+NAS+FPS-ZM1 group(n=8).The therapeutic effects of NAS were examined by hematoxylin-eosin(H&E)staining,and retinal ganglion cells(RGCs)counting.The expression of interleukin 1 beta(IL-1β),HMGB1,RAGE,and nod-like receptor 3(NLRP3)proteins and the phosphorylation of nuclear factorkappa B(p-NF-κB)were analyzed by immunohistochemistry staining and Western blot analysis.The expression of HMGB1 protein was also detected by enzyme-linked immunosorbent assay(ELISA).RESULTS:H&E staining results showed that NAS significantly reduced retinal edema and increased the number of RGCs in RIR rats.With NAS therapy,the HMGB1 and RAGE expression decreased significantly,and the activation of the NF-κB/NLRP3 pathway was antagonized along with the inhibition of p-NF-κB and NLRP3 protein expression.Additionally,NAS exhibited an anti-inflammatory effect by reducing IL-1βexpression.The inhibitory of RAGE binding to HMGB1 by RAGE inhibitor FPS-ZM1 led to a significant decrease of p-NF-κB and NLRP3 expression,so as to the IL-1βexpression and retinal edema,accompanied by an increase of RGCs in RIR rats.CONCLUSION:NAS may exhibit a neuroprotective effect against RIR via the HMGB1/RAGE/NF-κB signaling pathway,which may be a useful therapeutic target for retinal disease.

Effects of Melatonin on Spermatozoa Activity and Associated Mechanisms in Heat-Stressed Rats

In animals,heat stress(HS)disrupts spermatogenesis,reducing sperm quality and,in severe cases,potentially inducing the loss of male reproductive function.Melatonin confers significant resistance to oxidative stress and apoptosis;however,its specific effects on rat spermatocytes and the mechanism underlying its anti-HS effects remain inadequately explored.Therefore,this study aimed to analyze the effects of melatonin at different concentrations on sperm cell activity in heat-stressed rats.Modeling heat stress injury,sperm viability and density assay,sperm plasma membrane integrity analysis,and oxidative stress assay of testicular tissue were conducted.The results revealed that HS caused sperm cell injury.However,the intraperitoneal injection of melatonin effectively improved spermatozoa quality,and a dose of 1 mM significantly alleviated the HS-induced damage.Moreover,HS increased the levels of oxidative and endoplasmic reticulum(ES)stress in rat testicular tissues,inducing germ cell apoptosis and pathological changes.Similarly,melatonin treatment improved sperm cell viability and density,inhibited germ cell apoptosis,and reduced oxidative and ES stress levels.Overall,melatonin effectively reduced the adverse effects of HS on rat sperm cells,and an intraperitoneal injection of 1 mM(0.6966 mg)melatonin facilitated the normal production of spermatozoa.Notably,its mechanism may involve reduced ES and oxidative stress levels in testicular tissues,increased expression of the anti-apoptotic protein Bcl-2,and inhibition of germ cell apoptosis.