Abstract: Objective To explore the effects and mechanism of arglabin on neuroinflammation in mice with traumatic brain injury (TBI). Methods Adult male C57BL / 6J mice were divided randomly into: sham operation (Sham), Sham + Arglabin, TBI and TBI + Arglabin groups. TBI was induced by controlled cortical impact. After successful modeling, mice received 5 μg / kg arglabin by intraperitoneal injection, once a day until the material was retrieved. Neurological function was evaluated 3 weeks after the operation. Hematoxylin-eosin(HE) and Nissl staining were performed 4 weeks after the operation. Relative protein expression levels of NLRP3, apoptosis-associated specklike protein containing a caspase-recruitment domain ( ASC), and Caspase-1 in brain tissues were detected by Western Blot 3 d after surgery, and relative mRNA levels of the inflammatory factors interleukin (IL)-1β, IL-18, IL-6, inducible nitric oxide synthase ( iNOS), IL-4, and Arg1 were detected by RT-qPCR. M1 and M2 cells were observed 7 d after the operation. Results Compared with the findings in the Sham group, in TBI group mice, nerve function was reduced (P<0. 0001), the brain-tissue damage area was significantly increased ( P<0. 01), and neurons were largely lost (P<0. 01). Additionally, NLRP3, ASC, and Caspase-1 protein levels in brain tissue were significantly increased (P<0. 05), the proinflammatory cytokines IL-1β, IL-18, IL-6, and iNOS were significantly increased (P<0. 05), and the anti-inflammatory cytokines IL-4 and Arg1 were significantly decreased (P<0. 01).Lastly, levels of M1 type cells were significantly increased ( P<0. 01). Compared with the findings in the TBI group, in the TBI + Arglabin group, nerve function was significantly improved (P<0. 01), the brain tissue-damage area was significantly reduced ( P<0. 01), neuronal loss was significantly reduced ( P<0. 01), In addition,NLRP3, ASC, and Caspase-1 proteins in brain tissue were significantly reduced (P<0. 05), the proinflammatory cytokines IL-1β, IL-18, IL-6, and iNOS were significantly reduced (P<0. 05), the anti-inflammatory cytokines IL-4 and Arg1 were significantly increased (P<0. 01), and M2 type cells were significantly increased (P<0. 01). Conclusions Arglabin improves the local immune microenvironment by inhibiting NLRP3 inflammasome activation to alleviate neuroinflammation in mice with TBI.

Abstract: Objective To explore the effects of a Chinese medicine formula on regulating glucose, lipid, and cytokines in ZDF rats by transforming Qi and eliminating turbidity. Methods ZDF ( fa / fa) rats were divided randomly into diabetic (DM) and Chinese medicine-treated (TCM) groups (n= 8 per group) according to the levels of glucose after feeding with a high-fat diet. ZDF ( fa / +) rats were used as a control group ( NC, n= 8).“Qihuatongtiao formula”, which functions by transforming Qi and eliminating turbidity, was administered to rats in the TCM group for 6 weeks. Blood glucose levels were monitored and an oral glucose tolerance test was performed in the last week. Six weeks later, samples were collected and serum was retained for the detection of four lipid items and free fatty acids. Insulin levels were detected by ELISA. Pathological changes in the pancreas were observed by HE staining. Cytokine levels were measured using RayBio cytokine arrays. Results Compared with the NC group, the levels of blood glucose, four items of blood lipid, free fatty acid, insulin, and inflammatory cytokines (CXCL3, IL-2,IL-1α, etc. ) in the DM group were significantly increased, and the levels of blood glucose, four items of blood lipid,free fatty acid, insulin, and inflammatory cytokines in the TCM group were lower than those in the DM group.Degranulated islets were observed in the pancreas of rats in the DM group, while fewer degranulated islets were present in the TCM group. Conclusions The Chinese medicine “Qihuatongtiao formula”, which functions by transforming Qi and eliminating turbidity, could regulate glucose and lipids and alleviate inflammatory lesions in ZDF rats.

Abstract: Objective To investigate the efficacy of 3D-printed individualized titanium mesh ( 3D-PITM) combined with guided bone regeneration (GBR) in a rabbit model of osteoradionecrosis of the jaws (ORNJ) with bone defects. Methods Eighteen white rabbits with ORNJ-induced bone defects were divided randomly into three groups:a group (debridement and suturing for natural healing), b group (debridement and conventional GBR), and c group (debridement and 3D-PITM-supported GBR). Two rabbits from each group were euthanized at 4, 8, and 12 weeks,respectively. Bone defect repair was evaluated by immunohistochemical staining to detect expression levels of the osteogenesis-related factors alkaline phosphatase ( ALP ), osteocalcin ( OC), and bone morphogenetic protein-2 (BMP-2), and the inflammatory factors interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Results There was no significant difference in ALP expression among the groups at 4 weeks, but OC levels were lower in c group than in b group. At 8 and 12 weeks, however, ALP, BMP-2, and OC expression levels were all significantly higher in c group than in a and b groups. Expression levels of IL-6 and TNF-α were significantly lower in c group than in Groups a and b at 4, 8, and 12 weeks, but there was no significant difference in IL-6 expression levels between groups b and c at 4 weeks. Conclusions The combination of 3D-PITM and GBR demonstrated significantly superior bone defect repair efficacy in ORNJ compared with conventional GBR or natural healing alone.

Abstract: Objective To construct a forebrain excitatory neuron-specific NR4A1 knockout mouse model based on the LoxP-Cre recombinase system and to investigate the effects of this knockout on cognitive function and anxiety-like behaviors in mice. Methods NR4A1 ^{flox / +} mice were self-crossed and screened to obtain NR4A1 ^{flox / flox} mice, which were then crossed with Camk2a-Cre mice. NR4A1 ^{flox / +}-Camk2a-Cre F1 mice were obtained and further self-crossed. The result ing offspring were genotyped using tail genomic DNA to screen out NR4A1 ^{flox / flox}-Camk2a-Cre mice, which were defined as NR4A1 conditional knockout mice, specifically in forebrain excitatory neurons. Cognitive ability was assessed using behavioral tests, including novel object recognition and Y-maze tests, and anxiety-like behaviors were evaluated using the elevated plus maze, open field, and light-dark box tests. Expression levels of NR4A1 in the hippocampus, as a key forebrain region, were detected by Western Blot and immunofluorescence. Results ( 1 ) Genotyping confirmed that NR4A1 ^{flox / flox} -Camk2a-Cre forebrain excitatory neuron-specific NR4A1 knockout mice were successfully obtained. (2) NR4A1 protein levels were significantly reduced in the hippocampus in knockout mice compared with the findings in control mice, as shown by Western Blot. Also, the intensity of NR4A1 immunofluorescent signals in the hippocampus was also lower in knockout group compared with that in control group mice. (3) In cognitive behavioral tests, there was no significant difference in novel object recognition index, number of entries, distance traveled, or residence time in the novel arm of the Y-maze between knockout group and control group mice. (4) In anxiety-like behavior tests, knockout group mice spent more time in the central area of the open field than control group mice, but there was no significant difference in the distance traveled or number of entries into the central area. In the light-dark box test, knockout group mice spent longer in the light compartment than control group mice, but there was no significant difference in the distance traveled or number of entries into the light compartment between the two groups. There was no significant difference between knockout group and control group mice in the elevated plus maze test. Conclusions This study successfully constructed a forebrain excitatory neuronspecific NR4A1 knockout mouse model. Knockout mice showed no significant changes in cognitive level but a significant reduction in anxiety levels. This model provides an important experimental tool for in-depth investigations of the role of the NR4A1 gene in the physiological functions and pathological mechanisms of forebrain regions.

Abstract: Objective To explore the mechanism by which acupuncture wakes rats from traumatic brain injury (TBI) coma. Methods SD rats were divided randomly into blank, model, solvent control, acupuncture, and purinergic P2RX7 antagonist(anti-P2RX7)groups ( n= 12 rats per group). The model was established by controlled cortical impact. Rats in the solvent control, acupuncture, and anti-P2RX7 groups were injected with DMSO solution or P2RX7 antagonist in the periaqueductal gray ( vPAG) area of the midbrain for 30 min, respectively. After establishing the model, rats in the acupuncture and anti-P2RX7 groups were given acupuncture at Baihui and Shuigou points. Behavior was evaluated by coma time, state of consciousness, and modified neurological severity score (mNSS). The morphology of the vPAG area was observed by hematoxylin-eosin(HE) and Nissl staining, dopamine (DA) levels were detected by ELISA, and P2RX7 and dopamine transporter (DAT) expression were detected by immunofluorescence, Western Blot, and RT-qPCR. Results Compared with those in the blank group, rats in the model, solvent control, and anti-P2RX7 groups had prolonged coma durations, and increased consciousness and mNSS at each time point ( P<0. 05), increased pathological changes including neuronal degeneration, nuclear condensation, and Nissl’s corpuscle in the vPAG, and reduced DA, P2RX7, and DAT expressions in the vPAG (P<0. 05). Consciousness scores at 2. 5 and 4 hours after TBI, and mNSS at 12 hours after TBI were increased in the acupuncture group (P<0. 05), and pathological changes including neuron degeneration, nuclear condensation, and increased Nissl’s corpuscles existed in the vPAG. Compared with the findings in the model group, in the acupuncture group, the coma duration was shorter, the consciousness state and mNSS at each time point were decreased ( P<0. 05), tissue and neuron damage in the vPAG were reduced, and the DA level and P2RX7 and DAT expression were increased (P<0. 05). Compared with the findings in the solvent control group, in the acupuncture group, the coma duration was shortened, consciousness scores at 12, 24, and 48 h after TBI and the mNSS at 24 and 48 hours after TBI were decreased (P<0. 05), tissue and neuron injury in the vPAG were reduced, and DA, P2RX7, and DAT expression were increased (P<0. 05). Compared with the findings in the acupuncture group, in the anti-P2RX7 group, the coma duration was prolonged, the consciousness score and mNSS at each time point were increased (P<0. 05), tissue and neuron damage in the vPAG were increased, and DA, P2RX7, and DAT expression were decreased ( P<0. 05). Conclusions Acupuncture at GV20 and GV26 may regulate DA metabolism by upregulating P2RX7 expression, restoring pathological changes in the vPAG, improving the consciousness state and neurological function, and promoting coma awakening in rats after TBI.

Abstract: Objective To establish an animal model of co-infection with Mycoplasma pneumoniae and influenza virus H1N1. The aim is to clarify the co-pathogenic mechanism of co-infection of the two pathogens and provide a theoretical basis for the clinical intervention of mixed respiratory tract infections. Methods Twenty-four Syrian hamsters were divided into four groups: control ( C) group, infection group ( H1N1 influenza virus singleinfection(Flu) group, Mycoplasma pneumoniae single-infection(MP) group, and Mycoplasma pneumoniae and H1N1 influenza virus co-infection (F + M)group). The animals were challenged via intranasal drops and tracheal injection.Clinical indicators were recorded for 14 d, physiological parameters and pathogen shedding were monitored, and animals were selected on days 7 and 14 post-infection for histopathological examination, viral antigen detection, and nucleic acid testing. Results Syrian hamsters in Flu, MP and F + M group exhibited clinical symptoms similar to human influenza or Mycoplasma pneumoniae infections. Nucleic acids of both pathogens were detected in nasal swabs,throat swabs, anal swabs, blood, and tissues. Histopathological result showed varying degrees of pathological damage in different tissues of the infection groups. Conclusions The clinical symptoms, viral replication, and pathological manifestations confirm the successful establishment of a Syrian hamster model co-infected with Mycoplasma pneumoniae and influenza virus H1N1. This model provides an important experimental foundation for elucidating the mechanisms of respiratory co-infections, as well as for vaccine and drug development and clinical treatment strategies.

Abstract: Objective To establish and evaluate a sterile golden hamster diabetes model by fecal microbiota transplantation(FMT). Methods Twenty-five male SPF-grade SD rats were divided into the control (SD-CN) group and the model (SD-DM) group, with 10 rats in the SD-CN group and 15 rats in the SD-DM group. After being fed with normal feed and high-fat feed respectively for 3 weeks, the animals were fasted for 12 h. In the SD-DM group,Streptozotocin (STZ) was intraperitoneally injected at a dose of 25 mg / kg for 3 consecutive days. Twenty-four male golden hamsters were divided into a control (CN) group and a model (DM) group (n= 12 hamsters per group), the fecal suspensions of SD rats in the SD-CN group and the SD-DM group were inoculated respectively. The animals were euthanized 10 weeks after FMT and blood and small intestine, pancreas, and large intestine contents were collected for blood biochemistry, histopathological examination, and 16S rRNA sequencing. Results After fecal microbiota transplantation, apolipoprotein A1 and apolipoprotein B levels differed significantly between the two groups ( P<0. 05). Glucagon and fasting blood glucose levels were significantly increased (P<0. 01) and fasting insulin was significantly decreased (P<0. 001) in the DM group, presenting obvious characteristics of glycolipid metabolism disorder and insulin resistance. Compared with that in the CN group, the pancreas in the DM group showed multifocal fat deposition, islet atrophy, reduced islet cells, irregular islet shape, swelling and necrosis, islet cell proliferation,increased capillaries, and thickening of the basement membrane, demonstrated by hematoxylin-eosin(HE) staining.Hamsters in the DM group showed hyperplasia of the small intestinal villus epithelium, fusion and disordered arrangement of small intestinal villi, cell swelling accompanied by degeneration and necrosis, and aggregation of inflammatory cells in the lamina lining of the small intestinal mucosa. 16S rRNA sequencing showed significant differences in intestinal microbiota diversity and the abundance of specific microbiota between the two groups. The chao1, shannon, and simpson indexes of α diversity were significantly decreased in the DM group (P<0. 05) and the unweighted Unifrac distance was significantly decreased (P<0. 01), suggesting differences in β diversity. The composition of the intestinal bacterial communities differed significantly between the two groups, with significantly increased Verrucomicrobiota ( P<0. 05) and significantly decreased Firmicutes in the DM group ( P<0. 05). Conclusions A sterile golden hamster diabetes model was successfully established using fecal microbiota transplantation technology. The animals presented with clinical characteristics similar to those of diabetes, thus providing an animal model for in-depth research on the relationship between the intestinal microbiota and the occurrence and development of diabetes.

Abstract: Objective To study the qi-tonifying effects and mechanisms of different polar fractions of aqueous extract of Cinnamomum cassia branches and leaves in qi-deficiency rats, and to provide a reference for the pharmacodynamic material basis of Cinnamomum cassia qi-tonifying. Methods Ninety male SD rats were divided randomly into blank group, model group, positive group, aclministration group ( ethyl acetate fraction high dose group, ethyl acetate fraction low dose group, n-butanol fraction high dose group, n-butanol fraction low dose group,water fraction high dose group and water fraction low dose group) ( n= 10 rats per group). Except for the blank group, rats in the other groups were subjected to the “improper diet + fatigue” method to construct an animal model of qi deficiency. At the same time, rats were administered the fractions intragastrically once a day for 21 d. The body mass and weight-bearing swimming time were recorded after the last administration. Tumor necrosis factor-alpha (TNF-α), interleukin ( IL)-2, IL-12, creatine kinase ( CK), lactic acid ( LD), lactic dehydrogenase ( LDH),superoxide dismutase (SOD), malondialdehyde (MDA), and albumin (ALB) in serum, and adenosine diphosphate (ADP) and adenosine triphosphate ( ATP ) in liver were measured. Thymus, spleen, and liver indexes were measured, and the livers were observed by hematoxylin-eosin(HE) staining. Results Compared with the findings in the model group, qi-deficiency symptoms and liver tissue damage were improved in each administration group to different degrees. The ethyl acetate and n-butanol fractions significantly prolonged the weight-bearing swimming time in qi-deficient rats (P<0. 01). The high-dose ethyl acetate fraction significantly increased the thymus index and significantly improved TNF-α, IL-2, IL-12, CK, LD, LDH, SOD, MDA, ALB, ATP, and ADP levels (P<0. 05,P<0. 01), while the low-dose fraction significantly improved IL-2, LD, LDH, MDA, ALB, ATP, and ADP levels (P<0. 05, P<0. 01). The high-dose n-butanol fraction significantly increased the thymus index and significantly improved TNF-α, IL-2, IL-12, CK, LD, LDH, SOD, MDA, ALB, ATP, and ADP levels (P<0. 05, P<0. 01),and the low-dose fraction significantly improved IL-12, CK, LD, LDH, SOD, MDA, ALB, ATP, and ADP levels (P<0. 05, P<0. 01). The high-dose water fraction significantly improved IL-12, LD, LDH, MDA, and ATP levels ( P<0. 05, P<0. 01), while the low-dose fraction significantly improved LD levels ( P<0. 05 ). Conclusions Different polar fractions of the aqueous extract of Cinnamomum cassia branches and leaves can improve the symptoms of qi deficiency in rats to varying degrees. They can protect the thymus, spleen, and liver and regulate the levels of various factors in qi-deficiency rats, thereby improving their immunity and reducing organ damage.Through various effects such as qi supplementation, prolonging the time of fatigue, and enhancing the body’s exercise endurance, comprehensive indexes showed that the n-butanol fraction of the aqueous extract of Cinnamomum cassia branches and leaves had the greatest qi-invigorating effect.

Abstract:Estrogen receptors (ERs) are involved in regulating many complex physiological processes in the human body. Abnormal ERs signaling can lead to various diseases, including neurological disorders, osteoporosis,heart disease, and breast cancer. ERα, ERβ, and the novel G protein-coupled ER 1 (GPER-1) are considered to be the most important ERs. ERs knockout models can exhibit disease phenotypes and can serve as new animal models for studying ER-related diseases and their interventions. This paper reviews the research progress and important roles of estrogen and ERα, ERβ, and GPER-1, as well as the application of ERs knockout models in diseases.

Abstract:Microsatellite markers, composed of short tandem repeats of two to six nucleotides, are widely distributed in the genomes of eukaryotes. Their characteristics, including high levels of polymorphism, abundance,adherence to Mendelian inheritance laws, and ease of amplification via polymerase chain reaction, have made them the most commonly employed method for paternity testing in non-human primates. Abundant primate species represent invaluable genetic resources worldwide; however, the quantity and productivity of primate genetic resources have declined as a result of habitat destruction, illegal hunting, the pet trade, and inadequate protection and management.Research into primate conservation using molecular technologies has thus become a mainstream approach to protect primate species. This review systematically considers the structural characteristics and genetic properties of microsatellite markers and clarifies their advantages and limitations, thereby providing a theoretical basis for the rational selection of research tools in subsequent studies. We detail the significance of microsatellite markers in nonhuman primate research, as well as their applications in parentage identification in captive populations and the genetic assessment of wild populations. These applications demonstrate that microsatellite markers can accurately identify parentage, effectively avoid inbreeding, and safeguard population genetic diversity. They can also provide scientific data to support the formulation of species-conservation strategies through the assessment of indicators such as genetic diversity levels and gene flow. Finally, by integrating the current research status, this study discusses the existing limitations and challenges to the development of microsatellite markers, and the prospects for their future development by combining with next-generation sequencing and multi-omics technologies. This review thus provides scientific support for the population management of primates, the conservation of species diversity, and future related developments.

Abstract:Humanized hematopoietic stem cells ( Hu-HSCs ) provide a key model for simulating the differentiation and function of human T cells. The background of severe immune deficiency in reconstructed mice,however, means that thymic development is hindered or there is no thymic gland, lacking the site for T cell differentiation and maturation, leading to insufficient reconstruction of human functional T cells in mice. Although the transplantation of human fetal thymus tissue can support the effective development of T cells in humanized mice, the scarcity of donor tissues and ethical issues have limited its wide application. There is thus an urgent need to develop new types of thymus tissue to overcome the above limitations. This article systematically summarizes the basic characteristics of the humanized hematopoietic stem cells mouse model and the human T-cell immune response ability induced by traditional fetal thymus transplantation. We review the role of novel thymus transplantation in promoting T cell development and regulating the immune response mediated by human T cells, and reveal its potential mechanism and significance in enhancing human immunity. The result are expected to provide a theoretical basis and technical support for the development of new humanized mouse models.

Abstract:This study systematically reviewed the spectrum and applicability of rodent myocardial injury models, and proposes a selection and reporting framework of “ research question-mechanism-model-evaluationtranslation” to enhance their reproducibility and clinical translatability. Although numerous models have emerged,coronary artery ligation models that simulate myocardial infarction remain the cornerstone for investigating ischemic heart disease mechanisms and evaluating therapeutic strategies, because of their high fidelity in reproducing the pathophysiology, robust reproducibility, and stable replication of long-term outcomes such as ventricular remodeling.Considering ischemic, physical / chemical, pharmacological / inflammatory, and genetic models, we summarize the key procedural parameters, variable factors, and sources of bias, and integrate the morphological, functional, and molecular biomarker-based three-dimensional evaluation pathways. Permanent ligation is an appropriate strategy for studies of ventricular remodeling and chronic fibrosis, while ischemia-reperfusion closely mimics clinical reperfusion scenarios and reperfusion injury, and cryo- and thermal injury produce clearly demarcated focal lesions, facilitating repair, regeneration, and biomaterials evaluation. Isoproterenol-, doxorubicin-, and lipopolysaccharide-inducedmodels display phenotype specificity in relation to stress-induced necrosis, chemotherapy-related cardiotoxicity, and systemic inflammation, respectively, while genetic engineering models are advantageous for causal inference but involve higher costs and technical barriers. We recommend that models should be selected and combined according to the specific scientific questions, validated using stratified approaches, and accompanied by a minimal reporting checklist to reduce inter-experimental variability, strengthen the robustness of the evidence, and improve translational value.

Abstract:Endometriosis(EMs) affects both physical and mental health in women of reproductive age, but no clinical cure exists. Suitable animal models are essential to better understand the pathogenesis of endometriosis and inform clinical treatment. Rodents are currently the preferred choice because of their low cost, well-defined reproductive cycles, and short modeling periods. Conventional rodent models of endometriosis include autologous,allogeneic, and xenogeneic transplantation models. Researchers have refined these models to more accurately replicate human disease states and clinical manifestations. This review summarizes the major types of rodent models for endometriosis, factors that influence model development, and criteria used to evaluate success rates. It also discusses future directions for animal model advancement.