Abstract: Objective To construct collagen-induced arthritis ( CIA) rat models of different blood stasis syndrome types and evaluate various modeling method, thereby providing a basis for establishing stable and reliable rat models of blood stasis syndrome in rheumatoid arthritis (RA). Methods Based on the CIA rat model, six different blood stasis models were established by superimposing various factors including adrenaline injection, ice-water bath stimulation, starvation, lipopolysaccharide ( LPS) administration, and high-fat diet feeding. The modeling method were comprehensively evaluated by measuring joint swelling, arthritis index (AI) scoring, Micro-CT, hematoxylin and eosin (HE) staining, as well as assessing blood routine, hemodynamic parameters, and detecting inflammatory markers, coagulation indicators, and blood stasis-related indices via ELISA. Results Compared with the control group, the CIA-only group showed significantly increased inflammatory markers, hematological parameters,hemodynamic indices, and coagulation indicators. Micro-CT revealed significant bone destruction, and HE staining indicated rough articular surfaces and inflammatory cell infiltration. On this basis, different superimposed stimuli aggravated blood stasis-related manifestations to varying degrees: CIA + Adr group and CIA + LPS group exhibited comprehensive increases in inflammation, coagulation, and vasoactive substances. The CIA + Adr + ice water bath group showed worsened inflammation along with partial decreases in hemodynamic parameters. The CIA + Adr + ice water bath + starvation group demonstrated abnormalities in inflammatory and coagulation markers alongside reductions in certain factors. The CIA + LPS + high-fat diet group and CIA + high-fat diet group primarily exhibited abnormalities in hematological and vascular endothelial indicators. Conclusions Multiple stimuli can successfully induce different blood stasis syndrome types in CIA model rats, with certain differences observed. This study provides a comprehensive multidimensional evaluation of various modeling method for RA blood stasis syndrome, offering an experimental basis for further research into the pathogenesis and treatment strategies of this disease pattern.

Abstract: Objective To compare different dry eye mouse models induced by benzalkonium chloride solution, scopolamine hydrobromide subcutaneous injection, and a drying environment with and without benzalkonium chloride solution. Methods Fifty SPF C57BL / 6J mice were divided into five groups: mice in the normal control group received no intervention; mice in the scopolamine hydrobromide injection group were injected subcutaneously with scopolamine hydrobromide solution daily; mice in the benzalkonium chloride solution group received benzalkonium chloride eye drops daily; mice in the electric thermostatic drier group were maintained in a controlled dry environment; and mice in the combined benzalkonium chloridesolution / electric thermostatic drier group received benzalkonium chloride and were maintained in a controlled dry environment. Body mass, body temperature, behavioral changes, basic tear secretion-I test ( SIT), break-up time ( BUT), corneal fluorescein staining, and hematoxylin / eosin staining were compared before and after modeling. Apoptosis of corneal and lacrimal tissues was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling. Expression levels of nuclear factor-κB, interleukin-1β, tumor necrosis factor-α, and other inflammatory factors in lacrimal glands were detected by reverse transcription-quantitative polymerase chain reaction. Results After benzalkonium chloride modeling, the SIT and BUT results and body mass were significantly decreased ( P<0. 01) and body temperature, apoptosis, and inflammatory factors were all increased ( P<0. 01) compared with the normal control group. After scopolamine modeling, the SIT and BUT results were significantly reduced (P<0. 01), tension and anxiety were increased, the whole body and hair were dry, and stools were small and dry in the model compared with the normal control group.Field-experiment scores were increased (P<0. 05) and percentage of time spent in the OE% and TE% values in the elevated cross maze experiment were decreased (P<0. 01), and levels of inflammatory factors were increased (P<0. 05). Body mass gain decreased, body temperature increased (P<0. 01), SIT and BUT results were affected (P<0. 01 ), corneal injury was serious, tear secretion was significantly reduced, and inflammatory factors were increased (P<0. 05) in the combined benzalkonium chloride solution and electric thermostatic drier group. All the measured parameters were similar in the drying group compared with the normal control group. Conclusions Eye drops containing benzalkonium chloride, subcutaneous injection of scopolamine, and benzalkonium chloride plus a dry environment can induce dry eye model mice. Exposure to a drying environment alone for 28 days failed to induce a dry eye model, while scopolamine induced systemic xerosis rather than ocular surface symptoms, making it more suitable for establishing a mouse model of electric thermostatic drier. Benzalkonium chloride plus a dry environment produced the most stable and durable model, with typical clinical symptoms of a dry eye surface, plus high operational feasibility and good repeatability, making it an ideal model for teaching, and clinical and scientific research.

Abstract: Objective To investigate temporal relationships among neurotransmitter alterations, amyloid-beta (Aβ) deposition, and neuroinflammation in 5 × FAD mice. Methods 5 × FAD and C57BL / 6J (WT) mice at 1. 5 months (early pathological stage), 3 months (mid-pathological stage), and 6 months (late pathological stage) were examined to establish a time-window model of Alzheimer’s disease(AD) progression. Each age and genotype group included 8 mice. Behavioral phenotyping was assessed using the Y-maze, novel object recognition, and open field tests. Immunofluorescence was used to detect Aβ expression and glial cell activation in the hippocampus and prefrontal cortex (PFC). HE staining was performed to observe hippocampal neuronal morphology. Neurotransmitter levels in the hippocampus and PFC were quantified via high-performance liquid chromatography. Results At 1. 5 months, the hippocampal GABA level was lower in AD model group mice than in WT group mice. At 3 months,significant changes in several neurotransmitters, including acetylcholine, glutamate, serotonin, and 5- hydroxyindoleacetic acid (5-HIAA), were detected in the hippocampus of AD model group mice compared with WT group mice. At this stage, minor Aβ deposition and microglial activation were present in both the hippocampus and PFC. By 6 months, neurotransmitter levels further declined in the hippocampus and PFC of AD model group mice,accompanied by extensive Aβ deposition, pronounced hippocampal neuronal damage, and substantial glial cell activation. Cognitive impairments were also observed. Conclusions Neurotransmitter dysregulation in 5 × FAD mice emerged by 3 months of age and worsened with disease progression. These alterations were closely associated with Aβ deposition, neuroinflammation, and neuronal injury.

Abstract: Objective To establish a mouse model of temporal lobe epilepsy and to investigate the effects of the ketogenic diet (KD). Specifically, we aimed to compare the differences in body mass, fur condition, and blood ketone levels between mice on the KD and those on a regular diet; and to further explore the impact of KD on seizure susceptibility, as well as depression- and anxiety-like behaviors. Methods Thirty-one 4-weeks-old male SPF-grade ICR mice ( 20 ~ 22 g) underwent acclimatization for 1 week. Eleven mice were assigned randomly to control conventional diet (CON + ND, n= 5) and KD (CON + KD, n= 6) groups. The remaining 20 mice were subjected to pilocarpine-induced status epilepticus (SE). The SE model was validated using Racine scale scoring (≥ stage 4),and successfully modeled mice were allocated randomly to epileptic standard diet (SE + ND) and epileptic ketogenic diet group (SE + KD) groups. Seizure frequency was recorded via video monitoring, and anxiety- and depression-like behaviors were assessed using open field, elevated plus maze, and forced swimming tests ( FST). Results ( 1) Establishment of epilepsy models: of the 20 mice subjected to modeling, four failed to reach Racine stage 4 and two died. The remaining 14 successfully modeled mice were allocated randomly to SE + KD (n= 7) and SE + ND (n=7) groups. During intervention, two mice died in the SE + KD group, while no mortality occurred in the SE + ND group. (2)Establishment of the ketogenic diet model: epileptic mice had lower baseline body mass than controls, but there was no significant difference in weight change between the dietary interventions ( P>0. 05). Blood ketone levels in the CON + KD group were consistently ≥ 1. 0 mmol / L at days 1, 7, 14, and 28. By day 2 of ketogenic feeding, nine mice developed fur soiling with fecal matter and diarrhea, progressing to rectal prolapse in five severe cases by day 3. CON + ND and SE + ND mice retained glossy coats and formed stools throughout. (3) Behavioral tests: SE + KD mice had fewer seizures compared with SE + ND mice. In the FST, CON + KD exhibited shorter immobility times than CON + ND mice. Conclusions Standardized KD intervention effectively maintained therapeutic ketosis (blood ketones ≥ 1. 0 mmol / L) without significant weight impact. Notably, a KD suppressed epileptic discharges and ameliorated anxiety- and depression-like behaviors.

Abstract: Objective To establish a mouse model of temporal lobe epilepsy and to investigate the effects of the ketogenic diet (KD). Specifically, we aimed to compare the differences in body mass, fur condition, and blood ketone levels between mice on the KD and those on a regular diet; and to further explore the impact of KD on seizure susceptibility, as well as depression- and anxiety-like behaviors. Methods Thirty-one 4-weeks-old male SPF-grade ICR mice ( 20 ~ 22 g) underwent acclimatization for 1 week. Eleven mice were assigned randomly to control conventional diet (CON + ND, n= 5) and KD (CON + KD, n= 6) groups. The remaining 20 mice were subjected to pilocarpine-induced status epilepticus (SE). The SE model was validated using Racine scale scoring (≥ stage 4),and successfully modeled mice were allocated randomly to epileptic standard diet (SE + ND) and epileptic ketogenic diet group (SE + KD) groups. Seizure frequency was recorded via video monitoring, and anxiety- and depression-like behaviors were assessed using open field, elevated plus maze, and forced swimming tests ( FST). Results ( 1) Establishment of epilepsy models: of the 20 mice subjected to modeling, four failed to reach Racine stage 4 and two died. The remaining 14 successfully modeled mice were allocated randomly to SE + KD (n= 7) and SE + ND (n=7) groups. During intervention, two mice died in the SE + KD group, while no mortality occurred in the SE + ND group. (2)Establishment of the ketogenic diet model: epileptic mice had lower baseline body mass than controls, but there was no significant difference in weight change between the dietary interventions ( P>0. 05). Blood ketone levels in the CON + KD group were consistently ≥ 1. 0 mmol / L at days 1, 7, 14, and 28. By day 2 of ketogenic feeding, nine mice developed fur soiling with fecal matter and diarrhea, progressing to rectal prolapse in five severe cases by day 3. CON + ND and SE + ND mice retained glossy coats and formed stools throughout. (3) Behavioral tests: SE + KD mice had fewer seizures compared with SE + ND mice. In the FST, CON + KD exhibited shorter immobility times than CON + ND mice. Conclusions Standardized KD intervention effectively maintained therapeutic ketosis (blood ketones ≥ 1. 0 mmol / L) without significant weight impact. Notably, a KD suppressed epileptic discharges and ameliorated anxiety- and depression-like behaviors.

Abstract: Objective To investigate the effects of polysaccharides from wine-processed Polygonatum sibiricum (PSPW) on the behavioral performance and antioxidant indices of Alzheimer’s disease (AD) transgenic D.melanogaster, and evaluate its therapeutic effect on AD. Methods Amyloid β42(Aβ42 ) transgenic D. melanogaster were divided randomly into model, and low, medium, and high dose PSPW groups. w1118 D. melanogaster with the same genetic background as the control group. After 28 d of intervention, the lifespan, climbing ability, and olfactory memory were evaluated systematically. Aβ1-42 and malondialdehyde (MDA) levels in brain tissue, superoxide (SOD) and catalase (CAT) activities, and the expression levels of antioxidant genes in the Keap1 / Nrf2 signaling pathway were detected by ELISA and qPCR. Results Behavioral studies showed that AD transgenic D. melanogaster had a significantly shorter lifespan and significantly impaired climbing and olfactory memory abilities compared with the control group. PSPW significantly improved the above behavioral indicators compared with the model group.Regarding antioxidant indicators, brain tissue levels of Aβ1-42 and MDA levels were significantly increased in the model group, while SOD and CAT activities were significantly decreased. Medium and high dose PSPW group significantly reduced Aβ1-42 and MDA levels, increased SOD and CAT activities, and enhanced antioxidant capacity, while D. melanogaster in the low-dose PSPW group showed an improved but non-significant trend. Detection of antioxidant genes in the Kelch-like ECH-associated protein 1 (Keap1) / nuclear factor erythroid 2-related factor 2 (Nrf2) pathway showed that Keap1 expression was increased in the model group, while Nrf2, gamma-glutamylcysteine synthetase (GCL), and glutathione S-transferase S1 ( gsts1) expression were decreased. High dose PSPW group significantly down-regulated Keap1 and up-regulated Nrf2, GCL, and gsts1 expression, while the medium and low dose PSPW groups showed an activation trend but no significant difference. Conclusions PSPW regulate the Keap1 / Nrf2 antioxidant pathway, enhance antioxidant enzyme activities, reduce the accumulation of oxidative products, and achieve the therapeutic effects of reducing Aβ deposition in AD transgenic D. melanogaster and improving motor and memory dysfunctions.

Abstract: Objective To elucidate the metabolic reprogramming effects of Echinococcus multilocularis infection on host energy metabolism and its associated immunoregulatory mechanisms. Methods Serum levels of 57 central carbon metabolites in infected mice were detected using targeted metabolomics technology. The energy metabolic functions of peripheral blood mononuclear cell ( PBMC) and splenic CD4 ⁺T cells were evaluated via mitochondrial and glycolytic stress tests using the Seahorse XF24 energy metabolism analyzer system. Results Serum levels of L-arginine, succinate, isocitrate, dihydroxyacetone phosphate, and malate were significantly decreased (P<0. 05) in the infected group. Kyoto encyclopedia of genes and genomes analysis revealed significant alterations in five pathways, including central carbon metabolism in cancer. Both mitochondrial respiratory function and glycolytic capacity were suppressed in PBMCs in the infected group (P<0. 05). No significant mitochondrial dysfunction was observed in splenic CD4⁺ T cells, but their glycolytic activity was markedly reduced (P<0. 05). Conclusions E.multilocularis infection disrupts central carbon metabolism in mouse serum and significantly suppresses glycolytic function in both PBMCs and CD4⁺T cells. These findings suggest that energy metabolic reprogramming may play a critical role in parasite immune evasion.

Abstract: Objective To evaluate the biocompatibility, anti-calcification, and long-term stability of a novel balloon-expandable transcatheter aortic valve in pigs in vivo. Methods Twenty-seven healthy white pigs weighing approximately 45 kg were selected as experimental animals. Preoperative physical examination and appropriate pretreatment, including drug treatment and disinfection, were performed. Under aseptic surgical conditions, a valve loaded on the balloon catheter delivery system was implanted into the porcine aortic valve position via femoral artery vascular access. The animals’ vital signs, diet, and activity were then observed and continuous drug treatment was administered. Echocardiography and digital subtraction angiography were performed immediately and at 30, 90 and 180 d, to assess the hemodynamics and location of the valve, and the impact on the surrounding cardiac structures. Hearts and major organ tissues were sampled at the corresponding follow-up points, and the tissue inflammatory response, thrombus score, calcification degree, and endothelialization degree were analyzed by histopathology staining. The structural integrity of the valve stent and calcification of the valve leaflets were evaluated by X-ray and micro-computed tomography. Results Twenty-three animals were included in the final analysis. Pigs with successful valve implantation recovered well and had stable vital signs. Follow-up examination showed that the aortic valve blood flow velocity, transvalvular pressure difference, and leaflet motion were all normal or gradually improved, the stent position was stable, and the vessel remained unobstructed. Histological analysis showed that the tissue at the implantation site had mild inflammatory cell infiltration, no thrombosis, low calcification area, good stent integrity,and low calcification of the valve leaflets. All the result were in line with expectations, and the differences were statistically significant ( P<0. 05). Conclusions The new balloon-expandable transcatheter aortic valve system shows good biocompatibility, anti-calcification, and implant stability in animal experiments, laying a solid foundation for subsequent clinical trials. This system is expected to improve the therapeutic effect of transcatheter aortic valve implantation (TAVI) and promote the technological progress of domestic TAVI devices, to bring new breakthroughs and development opportunities in the field of cardiovascular disease treatment.

Abstract:Coronary microvascular disease ( CMVD) is a clinical syndrome characterized by myocardial ischemia caused by structural or functional abnormalities of the coronary precapillary arterioles, small arteries, and capillaries, resultsing from atherosclerotic or non-atherosclerotic pathogenic factors. Given the complex pathogenesis of CMVD, its increasing incidence, and the challenges in clinical diagnosis, it is crucial to develop animal models that can comprehensively reflect the disease’s characteristics and use these models to develop targeted therapeutic agents. Current approaches to constructing CMVD animal models primarily involve increasing hemodynamic load and disrupting metabolism to simulate secondary microvascular changes caused by diseases such as hypertension and diabetes, or directly inducing microvascular obstruction through surgical ligation of the coronary arteries or injection of thrombogenic agents. This article systematically reviews the characteristics of commonly used experimental animals in CMVD models, summarizes the method of model construction, the indications, and their advantages and disadvantages, to provide a reference for basic research in CMVD.

Abstract:Fecal microbiota transplantation ( FMT) is a therapeutic method in which microorganisms are transplanted from healthy donors’ feces into the gastrointestinal tract of patients, to directly change their intestinal microbiota, normalize its composition, and achieve therapeutic effects. Chronic diarrhea is closely associated with the gut microbiota and an imbalance in the gut microbiota can lead to the occurrence and progression of chronic diarrhea. FMT is an effective approach for the treatment of chronic diarrhea. This review summarizes the relationship between chronic diarrhea and the gut microbiota and considers research advancements and applications of FMT technology for managing chronic diarrhea. We also address the prevailing challenges and future prospects, to provide a reference for further studies of FMT for the treatment of chronic diarrhea.

Abstract:As one of the seven universal and five basic emotions, anger is a normal emotional response of the human body. When the external stimulation reaches certain thresholds, it affects both internal organs and psychology,which tends to induce a variety of unsettling changes. If the external stimulating factors are sudden, intense, and persistent, it may lead to systemic-potentially chronic-diseases. Animal models can simulate abnormal anger emotions of human beings caused by different inducements and can offer a basis for therapeutic studies of anger-related symptoms. Thus, these models provide important reference value for the treatment of diseases induced by anger,including aspects such as pathogenesis, drug development, and clinical efficacy evaluation. However, systematic research on the modeling method of anger is still lacking. Therefore, this article presents a review of the literature about anger expressed by aggressive behavior, the analysis included a combination of disease and syndrome in animal models and clinical research. The findings provide a reference for the design of anger-related models, mechanism research, and drug development.

Abstract:Primary biliary cholangitis ( PBC) is a prevalent autoimmune disease in middle-aged women,characterized by immune-mediated lymphocyte infiltration and bile duct destruction in intrahepatic bile ducts. The pathogenesis of PBC is complex, and its etiology remains unclear. Animal models can help to elucidate the pathogenesis and clinical treatment of PBC, but cannot fully simulate its clinical manifestations and pathological changes, even if the models show similar features of disease progression to those in humans. Here, we systematically compared the heterogeneity of PBC mouse models constructed using different induction method in terms of their genetic background, modeling principles, pathogenic cells, and immune characteristics. We then reviewed the differences in immune regulation and disease phenotypes among different PBC mouse models and explored the preclinical treatment strategies based on PBC mouse models. This work contributes to our understanding and the optimization of PBC mouse models, further elucidating the complex pathological processes of PBC and providing new insights for precise therapeutic interventions.

Abstract:Research on major sleep deprivation models has developed in recent years, both domestically and internationally, together with exploration of their applications in China’s four major chronic diseases. The main sleepdeprivation models in rats and mice include the modified multiple platform water environment, mild electric shock,running wheel, coplanar waveguide paradigm, sleep deprivation instrument, gentle stimulation, and sleep fragmentation method. The sleep deprivation models for zebrafish mainly include the light exposure, mild stimulation,and ecological water flow method, while the main models for Drosophila include the mechanical stimulation, repetitive light stimulation, and gene editing method. The four major chronic diseases, cardiovascular and cerebrovascular diseases, cancer, chronic respiratory diseases, and diabetes, have an increasingly significant impact on human health. Understanding the application of sleep deprivation models in research related to these four major chronic diseases provides a theoretical reference for animal models in future studies of these conditions.

Abstract:Alcoholic liver disease (ALD) has become one of the most important chronic liver diseases in China. In-depth studies of the pathogenesis and treatment strategies of ALD rely on the establishment of suitable animal models. This systematic review considers research progress in animal models of alcoholic liver disease,focusing on different construction method, the advantages and disadvantages and application value of commonly used animals, including rodents (such as the Lieber-De Carli, NIAAA/ Gao-Binge, and second strike models, et al) and zebrafish ( acute and chronic models), and briefly discusses emerging models such as miniature pigs and chicken embryos. This review thus aims to provide references for researchers to select suitable models, optimize existing models, and develop new animal models for alcoholic liver disease.

Abstract:Adipose-derived exosomes have emerged as important mediators of intercellular communication in the regulation of lipid metabolism, and their role has become a major focus of research in recent years. Lipid metabolism is closely associated with energy homeostasis and a wide range of metabolic diseases, including obesity,diabetes, and cardiovascular disorders. Adipose-derived exosomes may contribute to the maintenance of metabolic balance by regulating adipocyte secretory functions, energy metabolic pathways, and signaling between adipose tissue and other organs. They play particularly important roles in insulin resistance and adipose deposition, and they have been implicated in the development of metabolic diseases. Although multiple studies have investigated the potential mechanisms of adipose-derived exosomes, their functions in various physiological and pathological states remain unclear. This review summarizes recent research advances regarding adipose-derived exosomes in lipid metabolic diseases, with emphasis on their roles in adipocyte function, energy metabolism, and metabolic disease progression.These advances provide new perspectives for basic research and potential strategies for metabolic disease intervention.