Abstract: Objective　To establish and evaluate an integrated disease-syndrome rat model of chronic cerebral ischemia with Qi deficiency and blood stasis syndrome. Methods 　Thirty male Wistar rats were allocated randomly into three groups (n= 10 per group): sham operation (sham), 2-vessel occlusion (2-VO) group, and sleep deprivation combined with 2-VO (SD + 2-VO) group. We comprehensively assessed Qi deficiency and blood stasis syndrome manifestations in the rats using a dual evaluation approach, combining exhaustive swimming tests with quantitative tongue chroma analysis. Cognitive function was evaluated using the Barnes maze, and cerebral blood flow was compared using laser speckle contrast imaging. The histopathology of the hippocampal cytoarchitecture and white matter were examined using hematoxylin-eosin (HE) and Luxol fast blue (LFB) staining, respectively, and ultrastructural alterations of neurons in the hippocampal CA1 region were observed by transmission electron microscopy (TEM). Protein expression levels of NeuN, vascular endothelial growth factor A (VEGFA) and CD31 were detected by Western Blot and immunofluorescence. Results 　Cerebral blood flow was significantly reduced in rats in the 2-VO group compared with the sham group, but they failed to recapitulate the key clinical hallmarks of Qi deficiency and blood stasis syndrome. In contrast, rats in the SD + 2-VO group exhibited significantly reduced locomotor activity, exacerbated cerebral hypoperfusion, shortened swimming duration, and darkened tongue color compared with 2-VO rats. Rats in the SD + 2-VO group demonstrated significantly impaired learning and memory abilities in the Barnes maze test. Consistent with these observations, HE staining, TEM, and LFB staining revealed substantial neuronal and white matter damage in the SD + 2-VO group. NeuN expression was decreased and VEGFA and CD31 expression levels were increased in the 2-VO and SD + 2-VO groups, as shown by Western Blot. Taken together, these findings indicated that the SD + 2-VO model effectively recapitulated the clinical features of chronic cerebral ischemia with Qi deficiency and blood stasis pattern. Conclusions　The combination of sleep deprivation and bilateral carotid artery occlusion successfully established a rat model of chronic cerebral ischemia with Qi deficiency and blood stasis syndrome. Compared with the 2-VO model, SD + 2-VO model demonstrates more pronounced syndrome manifestations and better clinical relevance, thus providing a valuable animal model for traditional Chinese medicine research on chronic cerebral ischemia.

Abstract: Objective　To establish a mouse model of H1N1 influenza wind-heat syndrome by combining climate intervention with influenza virus nasal drops. Methods 　Seventy-two BALB/c mice were divided randomly into nine groups: a Control group, wind-heat(FR) groups (FR-3Day, FR-5Day), and Model groups (1LD-3Day, 2LD-3Day, 3LD-3Day, 1LD-5Day, 2LD-5Day, 2LD-5Day, 3LD-5Day) (n= mice per group). Mice in the Control group were housed in a normal environment, while mice in the FR and Model groups were kept in wind-heat conditions for 7 d. Mice in the Model groups received nasal PR8 influenza virus infection on the 8th day, and mice in the Control and FR heat groups received equal amounts of physiological saline nasal drops. After virus challenge, each group was housed in a normal environment and samples were taken on days 3 and 5. The appearance of the mice was observed and recorded and the lung index, routine blood parameters, lung tissue pathology, serum interleukin (IL)6 levels, and virus titers were detected in each group based on their behavioral status, stools, and body temperature. Results 　After 7 d of wind-heat intervention, mice in the FR groups showed no significant abnormalities in terms of appearance, stools, body temperature, routine blood parameters, or lung tissue pathology compared with the Control group. The appearance, lung index, red blood cell count, hemoglobin, hematocrit, pathological result, and body temperature in the Model groups worsened progressively with increasing time and toxin dosage, while the neutrophil percentage, lymphocyte percentage, virus titer, and serum IL-6 levels peaked on day 3 after viral attack, for the same viral dose, and then decreased slightly on day 5. Conclusions　PR8 nasal drops and 7 d of wind-heat climate intervention can be used to establish a mouse model of influenza wind-heat syndrome.

Abstract: Objective　To establish a mouse model of H1N1 influenza wind-heat syndrome by combining climate intervention with influenza virus nasal drops. Methods 　Seventy-two BALB/c mice were divided randomly into nine groups: a Control group, wind-heat(FR) groups (FR-3Day, FR-5Day), and Model groups (1LD-3Day, 2LD-3Day, 3LD-3Day, 1LD-5Day, 2LD-5Day, 2LD-5Day, 3LD-5Day) (n= mice per group). Mice in the Control group were housed in a normal environment, while mice in the FR and Model groups were kept in wind-heat conditions for 7 d. Mice in the Model groups received nasal PR8 influenza virus infection on the 8th day, and mice in the Control and FR heat groups received equal amounts of physiological saline nasal drops. After virus challenge, each group was housed in a normal environment and samples were taken on days 3 and 5. The appearance of the mice was observed and recorded and the lung index, routine blood parameters, lung tissue pathology, serum interleukin (IL)6 levels, and virus titers were detected in each group based on their behavioral status, stools, and body temperature. Results 　After 7 d of wind-heat intervention, mice in the FR groups showed no significant abnormalities in terms of appearance, stools, body temperature, routine blood parameters, or lung tissue pathology compared with the Control group. The appearance, lung index, red blood cell count, hemoglobin, hematocrit, pathological result, and body temperature in the Model groups worsened progressively with increasing time and toxin dosage, while the neutrophil percentage, lymphocyte percentage, virus titer, and serum IL-6 levels peaked on day 3 after viral attack, for the same viral dose, and then decreased slightly on day 5. Conclusions　PR8 nasal drops and 7 d of wind-heat climate intervention can be used to establish a mouse model of influenza wind-heat syndrome.

Abstract: Objective　To establish a rat model of venous thrombosis in a plateau hypobaric hypoxic environment and to investigate the effect of this environment on venous thrombosis. Methods 　A total of 144 healthy male SD rats were assigned randomly to four groups (n= 36 rats per group): a plains sham operation (A) group, plains operation (B) group, plateau altitude 6000 m + sham operation (C) group, and plateau altitude 6000 m + surgery (D) group. Rats in A and B groups were maintained in a plains normoxic environment, while rats in C and D groups C and D were subjected to a plateau environment. Rats in the surgical groups underwent quantitative constriction to incompletely obstruct the inferior vena cava blood flow. Each group was further divided into subgroups based on time: 1, 3, 5, 7, 14, and 21 d (n= 6 rats per group). Regular vascular ultrasound monitoring was conducted, and blood samples were taken for whole blood viscosity testing and the assessment of inflammatory indicators, including endothelin-1 (ET-1), interleukin-6 (IL-6) and tissue factor (TF). Coagulation function was evaluated through the activated partial thromboplastin time(APTT), prothrombin time(PT), thrombin time(TT), fibrinogen(FIB) and D-dimer. After the observation period, the experimental animals were sacrificed and the limbs were removed. Thrombus samples were stained with hematoxylin/eosin(HE), and the thrombus wet mass was measured. Results 　The thrombosis incidence was significantly higher in the plateau D group than in B group, accompanied by a marked increase in blood viscosity and hematocrit (P<.01). Additionally, levels of ET-1, IL-6, and TF were significantly elevated (P<.05), indicating a coagulation disorder. Conclusions　A plateau hypoxic environment model can be successfully simulated by quantitative coarctation of the inferior vena cava, combined with a specialized environmental chamber. The findings of this study suggest that a plateau hypoxic environment promotes venous thrombosis.

Abstract: Objective　To observe the differences in gut microbiota in chronic unpredictable mild stress (CUMS)-induced depression model rats of both sexes, and to provide experimental evidence for exploring sex differences in depression onset. Methods 　Thirty-two healthy SD rats were divided randomly into four groups based on sex: Male control group (Control-M), Female control group (Control-F), Male model group (Model-M), and Female model group (Model-F) (n= 8 rats per group). Rats in the control groups were fed without stimulation, while rats in the model groups were stimulated using the 28 d CUMS-induced depression method. After successful modeling, fresh feces were collected from all rats for high-throughput 16S rRNA sequencing. Behavioral observations were also conducted before and after preparing the model. Results 　The result of sucrose-preference, open-field, and forced-swimming tests differed significantly between the control and model groups. The result of the sucrose-preference test also differed between the sexes, while there was no difference in the open-field or forced-swimming test between the sexes. The α and β diversity of the gut microbiota genera showed an upward trend in the CUMS group compared with the control group. The ratio of Firmicutes/Bacteroidetes and the richness of the Roseburia and Lachnospiraceae_ NK4A136_group were decreased in male rats but showed an increasing trend in female rats. Conclusions　The ratio of Firmicutes/Bacteroidetes in the gut microbiota may be a key factor affecting the difference in the onset of depression between males and females, while the Roseburia and Lachnospiraceae_NK4A136_group be potential factors in correcting the gut microbiota and improving the symptoms of depression.

Abstract: Objective　To investigate the mechanism by which Yangmai Tongluo formula improves microcirculation disorders induced by high homocysteine (Hcy) levels via regulation of the acid sphingomyelinase (ASM) and endoplasmic reticulum (ER) stress pathways. Methods 　Fifty male C57BL/6J mice were divided randomly into a control group, Hcy model group, Yangmai Tongluo formula low-/high dose groups (5.3, 10.4 g/kg, respectively), and a folic acid group (0.08 g/kg). Except for the control group, microcirculation disorders were induced in all mice using drinking water containing 1.8 g/L Hcy for 6 weeks. After modeling for 2 weeks, mice were administered the corresponding treatments by gavage for 4 weeks. Serum Hcy concentrations and the blood perfusion volume of the lower extremity microvessels were measured. Protein expression levels of zonula occludens ZO-1, ZO-2, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), ASM, glucose regulated protein 78 (GRP78), and C/EBP homologous protein (CHOP) in the cardiac microvascular endothelium were analyzed using immunofluorescence. Results 　Serum Hcy levels were significantly increased in the Hcy model group compared with the control group (P<.05). Yangmai Tongluo formula did not significantly reduce Hcy levels compared with the Hcy model group, but blood perfusion in the lower extremities was significantly increased (P<.01) and expression levels of ZO-1 and ZO-2 in the cardiac microvascular endothelium were restored (P<.001) in the Yangmai Tongluo formula high dose group. It also inhibited the expression of ICAM-1, VCAM-1, ASM, GRP78, and CHOP (P<.05), with comparable effects to folic acid. Conclusions　Yangmai Tongluo formula improves Hcy-induced microcirculation disorders and endothelial dysfunction by inhibiting ASM activity and alleviating ER stress, via a mechanism closely related to the regulation of endothelial inflammation and barrier stability. These result provide experimental evidence to support the use of traditional Chinese medicine to treat microvascular diseases.

Abstract: Objective　To understand the current status of animal experimental research on comorbidities of depression, and to provide method for preparing and evaluating relevant animal models of comorbidities of depression. Methods 　We searched the CNKI and PubMed databases to identify research literature on comorbidities of depression in animals. We summarized and analyzed the data regarding animal species, comorbidity types, modeling method, and model evaluation indicators. Results 　This study included 100 Chinese and 143 English studies. The total number of papers published in China and elsewhere has shown an upward trend since 2011. The top three comorbidity models in terms of the number of papers published were depression pain (97, 39.91%), depression epilepsy (37, 15.23%), and depression diabetes (20, 8.23%). The depression neuropathic pain model was the most common comorbid type of depression and pain, and the most commonly used preparation method was selective injury of the sciatic nerve branch, often using C57BL/6J mice. The depression acute epilepsy model was the most common comorbid type of depression epilepsy, and was usually prepared using the lithium chloride pilocarpine model, often using SD rats. The most commonly used method for preparing the depression diabetes model was by intraperitoneal injection of streptozotocin, using C57BL/6J mice. We systematically summarized the evaluation method for the above comorbidity models. Conclusions　Increasing research is being carried out using animal models of comorbidities of depression. While a few animal models for the preparation and evaluation of comorbidities of depression are gradually progressing, most animal experimental studies on comorbidities of depression are still in their infancy, and further studies are required to explore these.

Abstract:Radiation-induced heart disease (RIHD) is a severe complication in patients with thoracic cancer undergoing radiotherapy, with important impacts on long-term survival among cancer survivors. There is an urgent need to investigate the pathogenesis of RIHD and to develop effective therapeutic agents, and the establishment of high-quality RIHD animal models is pivotal to addressing these issues. This review summarizes the critical factors to consider in establishing RIHD animal models, including species selection, radiation type, dosage, fractionation, and target fields, and modeling timeline, along with the evaluation method and success criteria. We also consider the potential pathogenic mechanisms underlying RIHD, including DNA damage, oxidative stress, inflammatory responses, mitochondrial dysfunction, renin-angiotensin-aldosterone system activation, and myocardial fibrosis, as well as their interrelationships. These insights provide a comprehensive reference framework for constructing RIHD animal models and advancing mechanistic investigations into this condition.

Abstract:Skeletal genetic diseases are characterized by disorders in bone development and growth throughout the body. Such diseases often present clinically with pathological manifestations such as head and limb deformities and scoliosis, which can seriously affect patient quality of life. The Nosology of genetic skeletal disorders: 2023 revision, recognizes 41 major categories, involving 552 genes; however, the pathogenic mechanisms of around half of all bone genetic diseases remain unclear. As a new type of model animal, zebrafish have a highly conserved skeletal development process and regulatory mechanism compared with mammals. They also have the advantages of small size, strong reproductive ability, short reproductive cycle, and transparent embryos, potentially making them suitable for studying the pathogenesis of human skeletal genetic diseases. This review focuses on progress in the application of zebrafish models in research related to human skeletal genetic diseases. We carried out an extensive literature review and selected nine major categories of skeletal genetic disorders for detailed discussion, including fibroblast growth factor receptor 3 chondrodysplasias, type 2 collagen disorders, and type 11 collagen disorders. This article summarizes the disease overview, zebrafish model construction, and their research significance, with the aim of providing a reference for in-depth research on the pathogenesis of human skeletal genetic diseases.

Abstract:Biomedical detection dogs (BMDD) can rely on their sensitive sense of smell to identify volatile organic compounds (VOCs) produced by lesions, enabling them to screen for various diseases in humans and plants and animals. Currently, there is limited research in this area domestically. To address this gap and enrich biomedical detection technology, a systematic review and analysis of the application of biomedical detection dogs in disease screening has been conducted. This exploration discusses the underlying scientific basis, current applications, key points, and future development trends. Research indicates that biomedical detection dogs have significant potential for development and application in screening for various diseases in humans, such as cancer, infectious diseases, and metabolic disorders, as well as in livestock, aquaculture diseases, and plant pests and diseases. They are expected to demonstrate greater economic value and social benefits in the future.

Abstract:Psoriasis is a chronic inflammatory skin disease with worldwide prevalence, primarily characterized by epidermal hyperplasia, abnormal keratinization, and immune cell infiltration, with a significant negative impact on patients’ quality of life and mental well-being. The onset of psoriasis is closely associated with genetic susceptibility, immune dysregulation, and environmental factors. Despite research progress into the pathogenesis of psoriasis, existing treatment method still face problems including limited efficacy and obvious side effects. There is thus an urgent need for an in-depth analysis of its pathological network and the development of novel interventional strategies. The imiquimod-induced psoriasis animal model has accordingly become a crucial tool for studying psoriasis owing to its high reproducibility and excellent pathological simulation. This review systematically summarizes the core mechanism of action of the imiquimod-induced psoriasis model, expounds on the molecular basis of its action via pathways such as the cascade reaction of the core immune-inflammatory axis, the multi-regulatory network of downstream synergistic mechanisms, and the interaction between host and environmental factors. Research based on this model has successfully verified the therapeutic effects of various targeted therapies and natural products on psoriasis, demonstrating its important application value in therapeutic interventional research. We also discuss the limitations of the imiquimod-induced psoriasis model, and indicate future research directions, with the aim of providing references for further in-depth research and the treatment of psoriasis.

Abstract:The global incidence rate of cancer is increasing yearly, and chemotherapy-induced gastrointestinal mucosal injury has become a crucial factor affecting patients’ therapeutic prognosis; however, there is currently a lack of effective therapeutic drugs to address this issue. There is thus an urgent need to establish more ideal animal models of chemotherapy-induced gastrointestinal mucosal injury, to support the exploration of its pathogenesis and the development of therapeutic drugs. This review considered relevant literature published during the period from 2019 to 2024, to provide a comprehensive summary and analysis from several perspectives, including the selection of experimental animals, chemotherapeutic drugs and modeling method, evaluation indicators, and practical applications. Furthermore, we highlight several existing issues with current models, including the lack of standardized modeling method, insufficient research on models with a tumor background, and inadequate exploration of novel cell death mechanisms. This collation of the literature also revealed the gradual emergence of traditional Chinese medicine as a research hotspot, with potential for the treatment of gastrointestinal mucosal injury. Further studies of effective medicines are warranted to identify interventional strategies for chemotherapy-induced gastrointestinal mucosal injury.