[in Chinese]
[in Chinese]
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 149 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.213
Standardization Challenges in Outcome Evaluation Systems of Animal Experiments and Considerations for Core Outcome Set Construction Strategies
ZHENG Qingyong, ZHOU Yongjia, LI Tengfei, XU Jianguo, TIAN Chen, LIU Hui, TIAN Min, ZHOU Ziyu, XU Caihua, CUI Yating, WANG Junfei, and TIAN Jinhui
Animal experimentation constitutes a critical link between basic research and clinical application, making its research quality and translational efficiency paramount. Although considerable progress has been made in standardizing operational procedures and ethical guidelines, the standardization of outcome evaluation systems has significantly lagged, creating a key bottleneck that constrains the quality of biomedical research and evidence synthesis. This deficiency is manifested by pronounced heterogeneity in outcome selection across similar studies, incomplete methodological reporting, and disparate criteria for result interpretation, which severely impairs the comparability of findings and the evidence integration. To cope with this challenge, this paper systematically introduces a mature methodological tool from clinical research–the core outcome set (COS)–and explores its construction strategies and application potential in the field of animal experimentation. Given the extensive diversity of animal experiments, a pragmatic strategy of "focusing on key areas, implementing phased pilots, and promoting gradual expansion" should be adopted. This approach prioritizes the development of domain-specific COS for disease areas characterized by high research volume, urgent translational needs, and well-established animal models. A multi-source integration pathway for COS development is detailed, comprising systematic literature searches, methodological appraisals, and expert consensus, with the feasibility of leveraging artificial intelligence (AI) to enhance efficiency also being examined. The development and promotion of such COS are not intended to restrict scientific exploration; rather, they aim to establish a new, tiered evaluation paradigm consisting of "core outcomes" (mandatory), "recommended outcomes" (encouraged), and "exploratory outcomes" (optional). This framework is expected not only to enhance research quality through standardization and to adhere to the "3R" principles but also to accelerate the accumulation of high-quality evidence. This, in turn, provides a solid foundation for higher-level evidence synthesis, ultimately facilitating the effective translation of basic research findings into clinical practice and providing an essential methodological framework for scientific advancement in relevant disciplines.Animal experimentation constitutes a critical link between basic research and clinical application, making its research quality and translational efficiency paramount. Although considerable progress has been made in standardizing operational procedures and ethical guidelines, the standardization of outcome evaluation systems has significantly lagged, creating a key bottleneck that constrains the quality of biomedical research and evidence synthesis. This deficiency is manifested by pronounced heterogeneity in outcome selection across similar studies, incomplete methodological reporting, and disparate criteria for result interpretation, which severely impairs the comparability of findings and the evidence integration. To cope with this challenge, this paper systematically introduces a mature methodological tool from clinical research–the core outcome set (COS)–and explores its construction strategies and application potential in the field of animal experimentation. Given the extensive diversity of animal experiments, a pragmatic strategy of "focusing on key areas, implementing phased pilots, and promoting gradual expansion" should be adopted. This approach prioritizes the development of domain-specific COS for disease areas characterized by high research volume, urgent translational needs, and well-established animal models. A multi-source integration pathway for COS development is detailed, comprising systematic literature searches, methodological appraisals, and expert consensus, with the feasibility of leveraging artificial intelligence (AI) to enhance efficiency also being examined. The development and promotion of such COS are not intended to restrict scientific exploration; rather, they aim to establish a new, tiered evaluation paradigm consisting of "core outcomes" (mandatory), "recommended outcomes" (encouraged), and "exploratory outcomes" (optional). This framework is expected not only to enhance research quality through standardization and to adhere to the "3R" principles but also to accelerate the accumulation of high-quality evidence. This, in turn, provides a solid foundation for higher-level evidence synthesis, ultimately facilitating the effective translation of basic research findings into clinical practice and providing an essential methodological framework for scientific advancement in relevant disciplines.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 138 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.056
Construction of Laboratory Animal Science and Technology Ethics Governance System in China and Its Preliminary Practice in Guangdong Province
LI Xiaoqin, YU Wenlan, DUAN Yizhu, LIU Zhonghua, WU Guodong, SHI Wenqi, and FU Hongkun
The welfare and ethics of laboratory animals are the ethical principles and behavioral norms that need to be followed in conducting animal-based scientific research, breeding and managing laboratory animals, and supervising and regulating such activities. The level of protection of laboratory animal welfare and ethics is closely related to the development of science and technology, which has become a widely recognized international consensus. At present, Guangdong Province is accelerating the construction of a high-level science and technology innovation province and the Guangdong-Hong Kong-Macao Greater Bay Area International Science and Technology Innovation Center. Guangdong Province should rely on its advanced governance capacity in the field of laboratory animal science and technology ethics to promote the high-quality development of its laboratory animal science and technology sector. Based on the management laws, regulations, and institutional mechanisms of laboratory animals in China, this paper explores the optimization of the laboratory animal science and technology ethics governance system, which includes the institutional guarantees, responsibility systems, ethical review and supervision mechanisms, and education and outreach. Through methods such as literature research, questionnaire surveys, and interview investigations, an empirical study of the laboratory animal science and technology ethics governance system in Guangdong Province has been conducted. Analysis of literature and research results shows that Guangdong Province has basically established a laboratory animal management system, collaboration mechanism, supervision mechanism, and education and training system that meet the current requirements of the laboratory animal science and technology ethics governance system in China. However, there are still problems such as an incomplete laboratory animal science and technology ethics supervision mechanism, an underdeveloped operation mechanism of review institutions, insufficient attention paid by laboratory animal units to the ethical review of animal experiments, inconsistent ethical review standards, and a lack of professional ethical education and training for ethics review personnel. Therefore, optimization measures such as improving the laboratory animal science and technology ethics review system, strengthening supervision and inspection, further strengthening the accountability of responsible entities, formulating review norms, and enhancing hierarchical and classified education and training are proposed, to provide a theoretical basis for promoting the normalized and long-term governance of laboratory animal science and technology ethics in Guangdong Province.The welfare and ethics of laboratory animals are the ethical principles and behavioral norms that need to be followed in conducting animal-based scientific research, breeding and managing laboratory animals, and supervising and regulating such activities. The level of protection of laboratory animal welfare and ethics is closely related to the development of science and technology, which has become a widely recognized international consensus. At present, Guangdong Province is accelerating the construction of a high-level science and technology innovation province and the Guangdong-Hong Kong-Macao Greater Bay Area International Science and Technology Innovation Center. Guangdong Province should rely on its advanced governance capacity in the field of laboratory animal science and technology ethics to promote the high-quality development of its laboratory animal science and technology sector. Based on the management laws, regulations, and institutional mechanisms of laboratory animals in China, this paper explores the optimization of the laboratory animal science and technology ethics governance system, which includes the institutional guarantees, responsibility systems, ethical review and supervision mechanisms, and education and outreach. Through methods such as literature research, questionnaire surveys, and interview investigations, an empirical study of the laboratory animal science and technology ethics governance system in Guangdong Province has been conducted. Analysis of literature and research results shows that Guangdong Province has basically established a laboratory animal management system, collaboration mechanism, supervision mechanism, and education and training system that meet the current requirements of the laboratory animal science and technology ethics governance system in China. However, there are still problems such as an incomplete laboratory animal science and technology ethics supervision mechanism, an underdeveloped operation mechanism of review institutions, insufficient attention paid by laboratory animal units to the ethical review of animal experiments, inconsistent ethical review standards, and a lack of professional ethical education and training for ethics review personnel. Therefore, optimization measures such as improving the laboratory animal science and technology ethics review system, strengthening supervision and inspection, further strengthening the accountability of responsible entities, formulating review norms, and enhancing hierarchical and classified education and training are proposed, to provide a theoretical basis for promoting the normalized and long-term governance of laboratory animal science and technology ethics in Guangdong Province.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 127 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.072
Analysis of Microbiological and Parasitic Testing Results for Laboratory Animals in Jiangxi Province, 2020-2024
XIAO Fangping, XIONG Lijuan, JIA Bo, LIU Huan, LUO Yongbing, and WAN Xiaorong
ObjectiveTo assess the current status of microbial and parasitic quality control for laboratory animals in Jiangxi Province by analyzing microbiological and parasitic test results from production facilities between 2020 and 2024, and to provide a basis for enhancing quality control measures.MethodsIn accordance with the current national standards for laboratory animals at the time of testing, the Jiangxi Provincial Laboratory Animal Quality Inspection Station (affiliated to Institute of Occupational Medicine of Jiangxi) conducted microbial and parasitic testing on 451 laboratory animals of 4 species from 6 laboratory animal production units in Jiangxi Province between 2020 and 2024, and analyzed the quality status of laboratory animals in the province.ResultsPasteurella pneumotropica was detected in one mouse sample in 2020, with a detection rate of 5.00%. Pseudomonas aeruginosa was detected in one mouse sample and mouse hepatitis virus antibody was detected in another mouse sample in 2023, with a detection rate of 2.78%, respectively. No microorganisms or parasites that should be excluded from SPF grade mice as specified in the national standards were detected in 2021, 2022, or 2024, with a qualification rate of 100.00%. Pasteurella pneumotropica was detected in four rat samples in 2020, with a detection rate of 20.00%. Pseudomonas aeruginosa was detected in two rat samples in 2021, with a detection rate of 10.00%, and Tyzzer's disease agent antibody was detected in four rat samples in 2024, with a detection rate of 10.00%. No microorganisms or parasites that should be excluded from SPF grade rats as specified in the national standards were detected in 2022 or 2023, with a qualification rate of 100.00%. For rabbits and guinea pigs, no microorganisms or parasites required to be tested for conventional grade rabbits and guinea pigs as specified in the national standards were detected from 2020 to 2024, with the qualification rate of both species reaching 100.00%.ConclusionBased on the microbial and parasitic testing results, the quality of rabbits and guinea pigs in Jiangxi Province is satisfactory. However, some issues persist with rats and mice. It is recommended to enhance the quality of experimental animals in Jiangxi Province by increasing the frequency of random inspections by quality testing units or by improving the self-inspection capabilities of production and user facilities.ObjectiveTo assess the current status of microbial and parasitic quality control for laboratory animals in Jiangxi Province by analyzing microbiological and parasitic test results from production facilities between 2020 and 2024, and to provide a basis for enhancing quality control measures.MethodsIn accordance with the current national standards for laboratory animals at the time of testing, the Jiangxi Provincial Laboratory Animal Quality Inspection Station (affiliated to Institute of Occupational Medicine of Jiangxi) conducted microbial and parasitic testing on 451 laboratory animals of 4 species from 6 laboratory animal production units in Jiangxi Province between 2020 and 2024, and analyzed the quality status of laboratory animals in the province.ResultsPasteurella pneumotropica was detected in one mouse sample in 2020, with a detection rate of 5.00%. Pseudomonas aeruginosa was detected in one mouse sample and mouse hepatitis virus antibody was detected in another mouse sample in 2023, with a detection rate of 2.78%, respectively. No microorganisms or parasites that should be excluded from SPF grade mice as specified in the national standards were detected in 2021, 2022, or 2024, with a qualification rate of 100.00%. Pasteurella pneumotropica was detected in four rat samples in 2020, with a detection rate of 20.00%. Pseudomonas aeruginosa was detected in two rat samples in 2021, with a detection rate of 10.00%, and Tyzzer's disease agent antibody was detected in four rat samples in 2024, with a detection rate of 10.00%. No microorganisms or parasites that should be excluded from SPF grade rats as specified in the national standards were detected in 2022 or 2023, with a qualification rate of 100.00%. For rabbits and guinea pigs, no microorganisms or parasites required to be tested for conventional grade rabbits and guinea pigs as specified in the national standards were detected from 2020 to 2024, with the qualification rate of both species reaching 100.00%.ConclusionBased on the microbial and parasitic testing results, the quality of rabbits and guinea pigs in Jiangxi Province is satisfactory. However, some issues persist with rats and mice. It is recommended to enhance the quality of experimental animals in Jiangxi Province by increasing the frequency of random inspections by quality testing units or by improving the self-inspection capabilities of production and user facilities.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 119 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.079
Experience Discussion on Feeding, Breeding and Genetic Management of Common Marmosets (Callithrix jacchus)
LI Li, LI Xuebo, FAN Yongheng, HE Donghua, LI Jianhong, and CHANG Liangtang
The common marmoset (Callithrix jacchus) is a species of the genus Callithrix in the primate family Callitrichidae. Common marmosets are an important scientific research resource for the study of neuroscience, medicine, and pharmacy. The feeding and breeding of common marmosets provide important support for scientific research. At present, common marmoset resources in China are at a critical stage of development. This paper takes the Center for Excellence in Brain Science and Intelligence Technology of the Chinese Academy of Sciences (hereafter referred to as CEBSIT) as an example to discuss the experience of common marmoset feeding, breeding and genetic management. In terms of breeding environment and facilities, on the basis of the national standard, more stringent environmental parameter standards are implemented for environmental temperature, relative humidity, the number of air exchanges, and the average concentration of sedimentation bacteria. Two kinds of cages for common marmoset breeding and experimentation are designed. According to the differences in facility conditions and staffing, a cleaning and maintenance method combining "dry rearing" and "wet rearing" is introduced. In terms of feeding management, in order to meet the high nutritional needs of common marmosets, CEBSIT prepares special formulated diets, and introduces the transition method of common marmosets from diets mixed with formulated rations and pellets to exclusive commercial diets, and strictly regulates the operation procedures of formulated diets for common marmosets, and has accumulated a series of methods for preventing nutritional and metabolic diseases in common marmosets. In terms of animal health management, the health of common marmosets is ensured through measures such as regular quarantine, isolation and treatment, and epidemic prevention, and the health inspection process of common marmosets is described in detail, and hierarchical and classified management is carried out according to the physical condition of common marmosets. This paper also briefly describes the treatment of diarrhea and pneumonia that are common in common marmosets. In terms of breeding management, it covers the selection of breeding common marmosets, combining pairs of common marmosets into the same cage for breeding, group splitting and hand-rearing of infant common marmosets, and introduces the method of reintroducing hand-reared infant common marmosets to parental care from the perspective of animal welfare. In terms of genetic management, the common marmoset population in CEBSIT is maintained as a closed colony and follows a non-inbreeding method. CEBSIT has developed an inbreeding screening system to check pedigree information before pairing to avoid inbreeding. Under conditions without introducing new individuals from outside, CEBSIT has continuously bred the colony for 5 generations while maintaining genetic heterozygosity. This paper only discusses the feeding, breeding and genetic management of the common marmoset and aims to provide a useful reference for other institutions engaged in common marmoset breeding.The common marmoset (Callithrix jacchus) is a species of the genus Callithrix in the primate family Callitrichidae. Common marmosets are an important scientific research resource for the study of neuroscience, medicine, and pharmacy. The feeding and breeding of common marmosets provide important support for scientific research. At present, common marmoset resources in China are at a critical stage of development. This paper takes the Center for Excellence in Brain Science and Intelligence Technology of the Chinese Academy of Sciences (hereafter referred to as CEBSIT) as an example to discuss the experience of common marmoset feeding, breeding and genetic management. In terms of breeding environment and facilities, on the basis of the national standard, more stringent environmental parameter standards are implemented for environmental temperature, relative humidity, the number of air exchanges, and the average concentration of sedimentation bacteria. Two kinds of cages for common marmoset breeding and experimentation are designed. According to the differences in facility conditions and staffing, a cleaning and maintenance method combining "dry rearing" and "wet rearing" is introduced. In terms of feeding management, in order to meet the high nutritional needs of common marmosets, CEBSIT prepares special formulated diets, and introduces the transition method of common marmosets from diets mixed with formulated rations and pellets to exclusive commercial diets, and strictly regulates the operation procedures of formulated diets for common marmosets, and has accumulated a series of methods for preventing nutritional and metabolic diseases in common marmosets. In terms of animal health management, the health of common marmosets is ensured through measures such as regular quarantine, isolation and treatment, and epidemic prevention, and the health inspection process of common marmosets is described in detail, and hierarchical and classified management is carried out according to the physical condition of common marmosets. This paper also briefly describes the treatment of diarrhea and pneumonia that are common in common marmosets. In terms of breeding management, it covers the selection of breeding common marmosets, combining pairs of common marmosets into the same cage for breeding, group splitting and hand-rearing of infant common marmosets, and introduces the method of reintroducing hand-reared infant common marmosets to parental care from the perspective of animal welfare. In terms of genetic management, the common marmoset population in CEBSIT is maintained as a closed colony and follows a non-inbreeding method. CEBSIT has developed an inbreeding screening system to check pedigree information before pairing to avoid inbreeding. Under conditions without introducing new individuals from outside, CEBSIT has continuously bred the colony for 5 generations while maintaining genetic heterozygosity. This paper only discusses the feeding, breeding and genetic management of the common marmoset and aims to provide a useful reference for other institutions engaged in common marmoset breeding.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 107 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.049
A Review of Methods for Establishing and Evaluating Animal Models of Stroke
YANG Yunrong, WU Wenyu, TAN Yue, YAN Guofeng, LI Yao, and LU Jin
Stroke is one of the leading causes of disability and mortality worldwide. Research into its mechanisms and the development of therapeutic strategies heavily rely on animal models that accurately replicate the pathological features of human disease. An ideal animal model for stroke should not only reproduce the neurological deficits and pathological changes observed in clinical patients but also demonstrate good reproducibility and translational value. This review focuses on the preparation and evaluation methods of ischemic stroke animal models. Firstly, it elaborates on the selection criteria, advantages, and disadvantages of experimental animals, including rodents (rats, mice) and non-rodents (non-human primates, miniature pigs, rabbits, zebrafish). Secondly, it provides a detailed overview of the modeling principles, key procedures, and application scopes for ischemic stroke models and hemorrhagic stroke models. Furthermore, the review summarizes advances in the applications of emerging technologies, including gene editing [e.g., clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing], multimodal imaging (e.g., two-photon microscopy, photoacoustic imaging), artificial intelligence, optogenetics, 3D bioprinting, organoid models, and multi-omics in model optimization, precise assessment, and mechanistic investigation. Finally, based on a systematic analysis of relevant domestic and international literature from 2019 to 2024, this review discusses model selection strategies based on research objectives, a multidimensional evaluation system encompassing behavioral, imaging, and molecular pathological assessments, and envisions future directions involving technological integration to achieve model precision and individualization. This article aims to provide a comprehensive methodological reference to help researchers select appropriate animal models of stroke according to specific scientific questions.Stroke is one of the leading causes of disability and mortality worldwide. Research into its mechanisms and the development of therapeutic strategies heavily rely on animal models that accurately replicate the pathological features of human disease. An ideal animal model for stroke should not only reproduce the neurological deficits and pathological changes observed in clinical patients but also demonstrate good reproducibility and translational value. This review focuses on the preparation and evaluation methods of ischemic stroke animal models. Firstly, it elaborates on the selection criteria, advantages, and disadvantages of experimental animals, including rodents (rats, mice) and non-rodents (non-human primates, miniature pigs, rabbits, zebrafish). Secondly, it provides a detailed overview of the modeling principles, key procedures, and application scopes for ischemic stroke models and hemorrhagic stroke models. Furthermore, the review summarizes advances in the applications of emerging technologies, including gene editing [e.g., clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing], multimodal imaging (e.g., two-photon microscopy, photoacoustic imaging), artificial intelligence, optogenetics, 3D bioprinting, organoid models, and multi-omics in model optimization, precise assessment, and mechanistic investigation. Finally, based on a systematic analysis of relevant domestic and international literature from 2019 to 2024, this review discusses model selection strategies based on research objectives, a multidimensional evaluation system encompassing behavioral, imaging, and molecular pathological assessments, and envisions future directions involving technological integration to achieve model precision and individualization. This article aims to provide a comprehensive methodological reference to help researchers select appropriate animal models of stroke according to specific scientific questions.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 94 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.037
Research Advances in Construction Methods and Novel Technologies for Animal Models of Pulmonary Hypertension
CHEN Ziyi, SUN Hongyan, KANG Pinfang, and WU Wenjuan
Pulmonary hypertension (PH), marked by sustained elevation of pulmonary artery pressure, imposes a heavy burden on the right ventricle and may culminate in right heart failure. Its pathogenesis is multifaceted, encompassing endothelial dysfunction, vascular smooth muscle proliferation, inflammation, thrombosis, and genetic factors. Animal models serve as core tools for exploring PH mechanisms and therapies, each with unique strengths and limitations. The single-dose monocrotaline (MCT) model is one of the most commonly used experimental animal models of PH and is widely applied in mechanistic studies, drug screening, and efficacy evaluation; it offers simplicity and cost-effectiveness, can induce PH within a short period, yet its pathophysiology differs to some extent from human idiopathic PH. In contrast, the Sugen5416 combined with chronic hypoxia model better mimics PH progression by placing animals under hypoxic conditions to induce pulmonary vasoconstriction and vascular remodeling, but it requires a longer modelling time, and the degree of hypoxia has a substantial impact on experimental outcomes. Beyond these two commonly used modeling approaches, a variety of emerging techniques have been applied in PH research; gene-editing technologies enable precise investigation of specific gene functions in PH. Additionally, induced pluripotent stem cell-based 3D organoid technology allows for individualized modelling while preserving patients' genetic information for precise clinical translation. Each model or technology can simulate different aspects of the pathological processes of human PH, and their findings provide key insights into the nature of the disease and serve as an important platform for the development of novel therapeutic targets. This paper comprehensively describes various animal models and emerging technologies used in PH research, analyzing their characteristics, applications, and limitations, with the aim of providing experimental and technical support for the development of new therapeutic strategies and drugs.Pulmonary hypertension (PH), marked by sustained elevation of pulmonary artery pressure, imposes a heavy burden on the right ventricle and may culminate in right heart failure. Its pathogenesis is multifaceted, encompassing endothelial dysfunction, vascular smooth muscle proliferation, inflammation, thrombosis, and genetic factors. Animal models serve as core tools for exploring PH mechanisms and therapies, each with unique strengths and limitations. The single-dose monocrotaline (MCT) model is one of the most commonly used experimental animal models of PH and is widely applied in mechanistic studies, drug screening, and efficacy evaluation; it offers simplicity and cost-effectiveness, can induce PH within a short period, yet its pathophysiology differs to some extent from human idiopathic PH. In contrast, the Sugen5416 combined with chronic hypoxia model better mimics PH progression by placing animals under hypoxic conditions to induce pulmonary vasoconstriction and vascular remodeling, but it requires a longer modelling time, and the degree of hypoxia has a substantial impact on experimental outcomes. Beyond these two commonly used modeling approaches, a variety of emerging techniques have been applied in PH research; gene-editing technologies enable precise investigation of specific gene functions in PH. Additionally, induced pluripotent stem cell-based 3D organoid technology allows for individualized modelling while preserving patients' genetic information for precise clinical translation. Each model or technology can simulate different aspects of the pathological processes of human PH, and their findings provide key insights into the nature of the disease and serve as an important platform for the development of novel therapeutic targets. This paper comprehensively describes various animal models and emerging technologies used in PH research, analyzing their characteristics, applications, and limitations, with the aim of providing experimental and technical support for the development of new therapeutic strategies and drugs.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 81 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.047
Literature Analysis and Validity Assessment for Animal Models of Attention Deficit and Hyperactive Disorder
LIAO Wangyue, LEI Shuang, LI Xuan, GUO Min, and ZHOU Ruoran
Attention deficit and hyperactive disorder (ADHD) is the most common neurodevelopmental disorder of childhood. It seriously impairs academic achievement, social interaction, and vocational development, and increases the risk of accidental injury and substance abuse. In some cases, the symptoms may also exert an indirect disruptive effect on public order. Its aetiology involves interactions among genetic, perinatal environmental, and psychosocial factors that cannot be fully disentangled by single clinical studies. Therefore, a systematic evaluation of existing animal models is essential for revealing pathophysiology and developing novel therapies. Using the keywords "attention deficit and hyperactive disorder", "models, animal", "validity", and their English equivalents, we systematically searched PubMed, Web of Science, CNKI, and Wanfang for publications from 2000 to 2025 (retrieving 328 publications) and added further references by citation tracking. Eighty-six rodent ADHD models that provided detailed construction protocols, behavioural assessments, neurobiological mechanisms, or pharmacological data were included and classified into spontaneous genetic, genetically engineered, and environmentally induced paradigms. Their face, construct, and predictive validity were compared. Among spontaneous genetic models, spontaneously hypertensive rats reproduce hyperactivity, impulsivity, and stimulant responses well, yet hypertension and sex differences limit specificity. Acallosal mouse strains link corpus callosum absence to ADHD-like behaviours, but neurotransmitter studies remain scarce. Genetically engineered rodents—including dopamine transporter, neurokinin-1 receptor and mediator complex subunit 23 knockout or conditional gene knockout lines—precisely dissect dopaminergic, noradrenergic, synaptic, or epigenetic pathways, yet generally lack full phenotypic coverage, social-deficit modelling, and comorbidity representation, and are accompanied by adverse effects such as growth retardation or ocular defects. Environmentally induced models employ lead, polychlorinated biphenyls, alcohol, nicotine exposures, 6-hydroxydopamine lesions, neonatal hypoxia, early social isolation, or maternal stress to recapitulate core symptoms. However, dose-schedule standardisation is lacking. Behavioural reversibility diverges from clinical persistence, and non-specific phenotypes such as anxiety or depression are common. Overall, no single paradigm simultaneously achieves high validity across all three dimensions. Currently, ADHD models have progressed from single-factor simulations to multidimensional evaluation, yet significant gaps remain in genetic-background standardisation, sex differences, cross-species translation, and syndrome-differentiation modelling under traditional Chinese medicine. Future directions should integrate genetic, environmental, and epigenetic interactions, establish life-span validation systems, and incorporate computational neuroscience alongside integrative Chinese-Western strategies to enhance clinical relevance and translational utility, thereby providing robust evidence-based support for mechanistic elucidation, drug screening and precision intervention in ADHD.Attention deficit and hyperactive disorder (ADHD) is the most common neurodevelopmental disorder of childhood. It seriously impairs academic achievement, social interaction, and vocational development, and increases the risk of accidental injury and substance abuse. In some cases, the symptoms may also exert an indirect disruptive effect on public order. Its aetiology involves interactions among genetic, perinatal environmental, and psychosocial factors that cannot be fully disentangled by single clinical studies. Therefore, a systematic evaluation of existing animal models is essential for revealing pathophysiology and developing novel therapies. Using the keywords "attention deficit and hyperactive disorder", "models, animal", "validity", and their English equivalents, we systematically searched PubMed, Web of Science, CNKI, and Wanfang for publications from 2000 to 2025 (retrieving 328 publications) and added further references by citation tracking. Eighty-six rodent ADHD models that provided detailed construction protocols, behavioural assessments, neurobiological mechanisms, or pharmacological data were included and classified into spontaneous genetic, genetically engineered, and environmentally induced paradigms. Their face, construct, and predictive validity were compared. Among spontaneous genetic models, spontaneously hypertensive rats reproduce hyperactivity, impulsivity, and stimulant responses well, yet hypertension and sex differences limit specificity. Acallosal mouse strains link corpus callosum absence to ADHD-like behaviours, but neurotransmitter studies remain scarce. Genetically engineered rodents—including dopamine transporter, neurokinin-1 receptor and mediator complex subunit 23 knockout or conditional gene knockout lines—precisely dissect dopaminergic, noradrenergic, synaptic, or epigenetic pathways, yet generally lack full phenotypic coverage, social-deficit modelling, and comorbidity representation, and are accompanied by adverse effects such as growth retardation or ocular defects. Environmentally induced models employ lead, polychlorinated biphenyls, alcohol, nicotine exposures, 6-hydroxydopamine lesions, neonatal hypoxia, early social isolation, or maternal stress to recapitulate core symptoms. However, dose-schedule standardisation is lacking. Behavioural reversibility diverges from clinical persistence, and non-specific phenotypes such as anxiety or depression are common. Overall, no single paradigm simultaneously achieves high validity across all three dimensions. Currently, ADHD models have progressed from single-factor simulations to multidimensional evaluation, yet significant gaps remain in genetic-background standardisation, sex differences, cross-species translation, and syndrome-differentiation modelling under traditional Chinese medicine. Future directions should integrate genetic, environmental, and epigenetic interactions, establish life-span validation systems, and incorporate computational neuroscience alongside integrative Chinese-Western strategies to enhance clinical relevance and translational utility, thereby providing robust evidence-based support for mechanistic elucidation, drug screening and precision intervention in ADHD.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 66 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.060
Regulation of Rat Intervertebral Disc Annulus Fibrosus Cell Proliferation and Apoptosis by Yaoshu Zhuyu Fang via miR-17-5P/MDM2/p53 Pathway
JIANG Haitao, YUAN Hantao, HUANG Wenting, YANG Rongrong, CHEN Xiaochun, YU Baoqing, and LI Sibo
ObjectiveTo investigate the effect of Yaoshu Zhuyu Fang on the regulation of the microRNA-17-5P (miR-17-5P)/murine double minute 2 (MDM2)/p53 axis in the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells, and its potential molecular mechanism.MethodsIntervertebral disc annulus fibrosus tissues were obtained from 8-week-old SPF-grade male SD rats, and annulus fibrosus cells were isolated and obtained by enzyme digestion and mechanical dispersion. Annulus fibrosus cells were divided into 6 groups: Group C was the blank control group, in which annulus fibrosus cells were not treated with interleukin-1 (IL-1) but were cultured in RPMI 1640 complete medium. Group was the degeneration model group constructed by treating annulus fibrosus cells with 10 ng/mL IL-1 for 24 h. Group +B was the IL-1 + blank serum group, in which annulus fibrosus cells were first treated with IL-1 to construct the degeneration model, then treated with RPMI 1640 medium containing 5% blank serum for 24 h. Group +W was the IL-1 + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1 to construct the degeneration model, then treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. Group +I was the IL-1 + miR-17-5P inhibitor group, in which annulus fibrosus cells were first treated with IL-1 to construct the degeneration model, then transfected with miR-17-5P inhibitor. Group +I+W was the IL-1 + miR-17-5P inhibitor + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1 to construct the degeneration model, then transfected with miR-17-5P inhibitor, and finally treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. CCK-8 assay was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis. Real-time quantitative PCR was used to detect the expression levels of miR-17-5P, MDM2 mRNA, and p53 mRNA in cells. Western blotting was used to detect the protein expression levels of MDM2 and p53 in cells. Dual-luciferase reporter system was used to analyze the targeting relationship between miR-17-5P and MDM2.ResultsCompared with Group C, Group showed a significant decrease in cell survival rate (P<0.001), a significant increase in cell apoptosis rate (P<0.001), significantly increased expression of miR-17-5P, p53 mRNA, and p53 protein (P<0.001), and significantly decreased expression of MDM2 mRNA and protein (P<0.001). Compared with Group , Group +W, Group +I, and Group +I+W showed significantly increased cell survival rate, significantly decreased apoptosis rate, significantly decreased expression of miR-17-5P, p53 mRNA, and p53 protein, and significantly increased expression of MDM2 mRNA and protein (P<0.001). Moreover, changes in the above indicators were greater in Group +I+W (P<0.001). Circular RNA Interactome predicted that miR-17-5P had specific binding sites with the 3' untranslated region (3'UTR) of MDM2. Transfection of miR-17-5P mimic significantly reduced the luciferase expression level of co-transfected luciferase reporter plasmid containing wild-type MDM2 3'UTR (P<0.05), but had no significant effect on luciferase expression in cells co-transfected with luciferase reporter plasmid containing mutant MDM2 3'UTR (P>0.05).ConclusionYaoshu Zhuyu Fang down-regulates the expression of miR-17-5P, promotes the synthesis of MDM2 protein, thereby down-regulates p53, promotes proliferation, and inhibits the apoptosis of rat intervertebral disc annulus fibrosus cells.ObjectiveTo investigate the effect of Yaoshu Zhuyu Fang on the regulation of the microRNA-17-5P (miR-17-5P)/murine double minute 2 (MDM2)/p53 axis in the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells, and its potential molecular mechanism.MethodsIntervertebral disc annulus fibrosus tissues were obtained from 8-week-old SPF-grade male SD rats, and annulus fibrosus cells were isolated and obtained by enzyme digestion and mechanical dispersion. Annulus fibrosus cells were divided into 6 groups: Group C was the blank control group, in which annulus fibrosus cells were not treated with interleukin-1β (IL-1β) but were cultured in RPMI 1640 complete medium. Group β was the degeneration model group constructed by treating annulus fibrosus cells with 10 ng/mL IL-1β for 24 h. Group β+B was the IL-1β + blank serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% blank serum for 24 h. Group β+W was the IL-1β + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. Group β+I was the IL-1β + miR-17-5P inhibitor group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor. Group β+I+W was the IL-1β + miR-17-5P inhibitor + Yaoshu Zhuyu Fang-containing serum group, in which annulus fibrosus cells were first treated with IL-1β to construct the degeneration model, then transfected with miR-17-5P inhibitor, and finally treated with RPMI 1640 medium containing 5% Yaoshu Zhuyu Fang-containing serum for 24 h. CCK-8 assay was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis. Real-time quantitative PCR was used to detect the expression levels of miR-17-5P, MDM2 mRNA, and p53 mRNA in cells. Western blotting was used to detect the protein expression levels of MDM2 and p53 in cells. Dual-luciferase reporter system was used to analyze the targeting relationship between miR-17-5P and MDM2.ResultsCompared with Group C, Group β showed a significant decrease in cell survival rate (P<0.001), a significant increase in cell apoptosis rate (P<0.001), significantly increased expression of miR-17-5P, p53 mRNA, and p53 protein (P<0.001), and significantly decreased expression of MDM2 mRNA and protein (P<0.001). Compared with Group β, Group β+W, Group β+I, and Group β+I+W showed significantly increased cell survival rate, significantly decreased apoptosis rate, significantly decreased expression of miR-17-5P, p53 mRNA, and p53 protein, and significantly increased expression of MDM2 mRNA and protein (P<0.001). Moreover, changes in the above indicators were greater in Group β+I+W (P<0.001). Circular RNA Interactome predicted that miR-17-5P had specific binding sites with the 3' untranslated region (3'UTR) of MDM2. Transfection of miR-17-5P mimic significantly reduced the luciferase expression level of co-transfected luciferase reporter plasmid containing wild-type MDM2 3'UTR (P<0.05), but had no significant effect on luciferase expression in cells co-transfected with luciferase reporter plasmid containing mutant MDM2 3'UTR (P>0.05).ConclusionYaoshu Zhuyu Fang down-regulates the expression of miR-17-5P, promotes the synthesis of MDM2 protein, thereby down-regulates p53, promotes proliferation, and inhibits the apoptosis of rat intervertebral disc annulus fibrosus cells.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 55 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.043
Establishment and Evaluation of an Oxidative Stress Model of Atopic Dermatitis Induced by 2,4-dinitrofluorobenzene
LIU Chang, XIANG Xuesong, HE Huihuang, CHEN Xiaoqing, and QIU Wenhong
ObjectiveTo establish an oxidative stress mouse model of atopic dermatitis (AD) by applying 2,4-dinitrofluorobenzene (DNFB) to the back and post-auricular skin of KM mice, and to evaluate the regulatory role of the RAGE-NLRP3 axis (receptor for advanced glycation end products-NOD-like receptor family, pyrin domain containing 3 axis) in AD-related oxidative stress, thereby providing a potential therapeutic target for AD treatment.MethodsTwenty SPF-grade female KM mice were randomly divided into a control group (Control group) and an experimental group (DNFB group), with 10 mice in each group. Mice in the Control group were treated with an acetone-olive oil vehicle (acetone: olive oil = 3: 1) on their back and post-auricular skin. Mice in the DNFB group were treated with 0.5% DNFB (prepared by adding 0.5 g DNFB per 100 mL of acetone-olive oil vehicle) on the same areas, once daily for 14 consecutive days. The severity of skin lesions was scored on days 2, 4, 6, 9, 12, and 14 of treatment. On day 14, scratching behavior and ear thickness were evaluated. Ear swelling was evaluated on the final day by measuring bilateral ear thickness three times with a vernier caliper; the three measurements were averaged. HE staining was used to observe morphological and structural changes of cells in the back skin tissues. The mRNA and protein expression levels of RAGE (receptor for advanced glycation end products) in skin tissues were detected by quantitative real-time PCR, Western blot, and immunohistochemical staining. The mRNA expression levels of oxidative stress-related molecules, including NLRP3 (NOD-like receptor family, pyrin domain containing 3), caspase-1 (cysteine-dependent aspartate-specific protease 1), and IL-1 (Interleukin-1), were detected by quantitative real-time PCR.ResultsOn day 14, the back skin lesion scores of the Control group and DNFB group were (0.20±0.42) and (9.93±1.30) (P<0.000 1), respectively. Scratching behavior scores were (5.00±2.05) and (49.26±8.49) episodes, respectively (P<0.000 1), and ear thicknesses were (213.00±11.87) μm and (765.93±140.47) μm (P<0.000 1), respectively. The DNFB group exhibited marked skin dryness, desquamation, and thickening. HE staining results showed that skin inflammation was obvious in the DNFB group, consistent with the pathological features of AD. Quantitative real-time PCR and Western blot results showed that compared with the Control group, the mRNA expression level of RAGE in skin tissues of the DNFB group was significantly increased (P<0.05), and the protein expression level of RAGE was also significantly increased (P<0.01). Immunohistochemical staining results showed that compared with the Control group, skin tissue sections of the DNFB group exhibited thickened stratum corneum and fibrotic proliferation of fibroblasts in the interstitium under microscopic observation, with a significant increase in RAGE protein expression in the skin tissues (P<0.01). Quantitative real-time PCR results showed that the mRNA expression levels of NLRP3, caspase-1, and IL-1 in skin tissues of the DNFB group were all significantly increased (P<0.01).ConclusionThe AD mouse oxidative stress model has been successfully established by topical DNFB application. RAGE may promote the development of AD by regulating the NLRP3 inflammasome and IL-1 release, forming an oxidative-inflammatory cascade, suggesting that it could be a potential therapeutic target for AD.ObjectiveTo establish an oxidative stress mouse model of atopic dermatitis (AD) by applying 2,4-dinitrofluorobenzene (DNFB) to the back and post-auricular skin of KM mice, and to evaluate the regulatory role of the RAGE-NLRP3 axis (receptor for advanced glycation end products-NOD-like receptor family, pyrin domain containing 3 axis) in AD-related oxidative stress, thereby providing a potential therapeutic target for AD treatment.MethodsTwenty SPF-grade female KM mice were randomly divided into a control group (Control group) and an experimental group (DNFB group), with 10 mice in each group. Mice in the Control group were treated with an acetone-olive oil vehicle (acetone: olive oil = 3: 1) on their back and post-auricular skin. Mice in the DNFB group were treated with 0.5% DNFB (prepared by adding 0.5 g DNFB per 100 mL of acetone-olive oil vehicle) on the same areas, once daily for 14 consecutive days. The severity of skin lesions was scored on days 2, 4, 6, 9, 12, and 14 of treatment. On day 14, scratching behavior and ear thickness were evaluated. Ear swelling was evaluated on the final day by measuring bilateral ear thickness three times with a vernier caliper; the three measurements were averaged. HE staining was used to observe morphological and structural changes of cells in the back skin tissues. The mRNA and protein expression levels of RAGE (receptor for advanced glycation end products) in skin tissues were detected by quantitative real-time PCR, Western blot, and immunohistochemical staining. The mRNA expression levels of oxidative stress-related molecules, including NLRP3 (NOD-like receptor family, pyrin domain containing 3), caspase-1 (cysteine-dependent aspartate-specific protease 1), and IL-1β (Interleukin-1β), were detected by quantitative real-time PCR.ResultsOn day 14, the back skin lesion scores of the Control group and DNFB group were (0.20±0.42) and (9.93±1.30) (P<0.000 1), respectively. Scratching behavior scores were (5.00±2.05) and (49.26±8.49) episodes, respectively (P<0.000 1), and ear thicknesses were (213.00±11.87) μm and (765.93±140.47) μm (P<0.000 1), respectively. The DNFB group exhibited marked skin dryness, desquamation, and thickening. HE staining results showed that skin inflammation was obvious in the DNFB group, consistent with the pathological features of AD. Quantitative real-time PCR and Western blot results showed that compared with the Control group, the mRNA expression level of RAGE in skin tissues of the DNFB group was significantly increased (P<0.05), and the protein expression level of RAGE was also significantly increased (P<0.01). Immunohistochemical staining results showed that compared with the Control group, skin tissue sections of the DNFB group exhibited thickened stratum corneum and fibrotic proliferation of fibroblasts in the interstitium under microscopic observation, with a significant increase in RAGE protein expression in the skin tissues (P<0.01). Quantitative real-time PCR results showed that the mRNA expression levels of NLRP3, caspase-1, and IL-1β in skin tissues of the DNFB group were all significantly increased (P<0.01).ConclusionThe AD mouse oxidative stress model has been successfully established by topical DNFB application. RAGE may promote the development of AD by regulating the NLRP3 inflammasome and IL-1β release, forming an oxidative-inflammatory cascade, suggesting that it could be a potential therapeutic target for AD.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 46 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.044
Research on Spatiotemporal Gene Expression Profiles and Repair Mechanisms of Spinal Cord Compression and Hemisection Spinal Cord Injury Mouse Models
XU Bo, CHEN Tairen, FANG Qian, and WU Ji
ObjectiveTo investigate the gene expression sequence and molecular mechanisms in the local microenvironment during the subacute to chronic phases (1-28 days) in mouse models of spinal cord compression injury and hemisection spinal cord injury, thereby revealing the molecular characteristics of spinal cord repair and providing a theoretical basis for selecting therapeutic targets for spinal cord injury.MethodsThirty-six 8-9-week-old SPF-grade ICR mice were randomly divided into three groups (n=12 per group): sham-operated control (CTR) group, hemisection spinal cord injury (HSCI) group, and spinal cord compression injury (SCC) group. Mice in the CTR group underwent the same surgical preparation and anesthesia, followed by a dorsal midline incision at the T9-T10 segment. After layer-by-layer dissection and removal of the corresponding lamina, the spinal cord dura mater was fully exposed and kept intact. The cord was exposed to air for 10 minutes (matching the duration of the compression injury group), during which any instrument contact with the cord was avoided. The incision was then irrigated and sutured. The HSCI group underwent a 70% transection of the T9 spinal cord segment using micro-instruments to establish a hemisection spinal cord injury model. The SCC group underwent sustained compression of the T10 spinal cord segment for 10 minutes using a self-made compressor (a 30 g solid small iron bar) to establish a spinal cord compression injury model. Motor function recovery was assessed using the modified Basso-Beattie-Bresnahan (BBB) score on postoperative days 1, 3, 7, 14, 21, and 28. On days 7 and 14 post-operation, mice were anesthetized, and the injured spinal cord segments were harvested. The evolution of specific molecular networks in the spinal cord injury mouse models was analyzed via RNA sequencing (RNA-Seq) and enrichment analysis, and the expression of key genes was verified using real time fluorogenic quantitative PCR.ResultsBBB scores indicated that motor function recovery in the SCC group was significantly better than that in the HSCI group, with BBB scores showing a continuously increasing trend and remaining higher than those in the HSCI group over the 4-week period (P<0.001). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses based on RNA-Seq differentially expressed genes revealed that, compared to the CTR group, genes related to the extracellular matrix were significantly up-regulated (P<0.05), while genes related to axon guidance were significantly down-regulated (P<0.05) in the SCC group on day 7 post-operation. On day 21, genes involved in immune regulation and the retinol signaling pathway were significantly activated in the SCC group (P<0.05). In contrast, in the HSCI group, genes associated with inflammation and immune response were significantly up-regulated (P<0.001), while genes related to neuronal differentiation and synapse formation were significantly down-regulated (P<0.001) on day 7. On day 21, genes related to cell-matrix junctions and N-methyl-D-aspartate receptors were significantly up-regulated (P<0.001) in the HSCI group. Furthermore, compared to the SCC group, the HSCI group exhibited different pathway enrichment characteristics in GO and KEGG analyses on days 7 and 21 post-injury. On day 7, genes involved in the NOD-like receptor signaling pathway and the complement and coagulation cascades were significantly up-regulated in the HSCI group (P<0.001). On day 21, genes related to the extracellular matrix-receptor interaction and the neuroactive ligand-receptor interaction pathways were significantly activated (P<0.001). Finally, real time fluorogenic quantitative PCR validation results were highly consistent with the RNA-Seq results, further confirming the differential expression trends of key genes between the SCC and HSCI groups.ConclusionThe SCC and HSCI injury models may drive distinct repair pathways: the preservation of some axons in the SCC model predisposes it toward tissue repair, whereas the HSCI model requires the coordination of more complex molecular networks to achieve a new equilibrium. This finding further deepens the understanding of the heterogeneous regulatory mechanisms underlying spinal cord injury.ObjectiveTo investigate the gene expression sequence and molecular mechanisms in the local microenvironment during the subacute to chronic phases (1-28 days) in mouse models of spinal cord compression injury and hemisection spinal cord injury, thereby revealing the molecular characteristics of spinal cord repair and providing a theoretical basis for selecting therapeutic targets for spinal cord injury.MethodsThirty-six 8-9-week-old SPF-grade ICR mice were randomly divided into three groups (n=12 per group): sham-operated control (CTR) group, hemisection spinal cord injury (HSCI) group, and spinal cord compression injury (SCC) group. Mice in the CTR group underwent the same surgical preparation and anesthesia, followed by a dorsal midline incision at the T9-T10 segment. After layer-by-layer dissection and removal of the corresponding lamina, the spinal cord dura mater was fully exposed and kept intact. The cord was exposed to air for 10 minutes (matching the duration of the compression injury group), during which any instrument contact with the cord was avoided. The incision was then irrigated and sutured. The HSCI group underwent a 70% transection of the T9 spinal cord segment using micro-instruments to establish a hemisection spinal cord injury model. The SCC group underwent sustained compression of the T10 spinal cord segment for 10 minutes using a self-made compressor (a 30 g solid small iron bar) to establish a spinal cord compression injury model. Motor function recovery was assessed using the modified Basso-Beattie-Bresnahan (BBB) score on postoperative days 1, 3, 7, 14, 21, and 28. On days 7 and 14 post-operation, mice were anesthetized, and the injured spinal cord segments were harvested. The evolution of specific molecular networks in the spinal cord injury mouse models was analyzed via RNA sequencing (RNA-Seq) and enrichment analysis, and the expression of key genes was verified using real time fluorogenic quantitative PCR.ResultsBBB scores indicated that motor function recovery in the SCC group was significantly better than that in the HSCI group, with BBB scores showing a continuously increasing trend and remaining higher than those in the HSCI group over the 4-week period (P<0.001). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses based on RNA-Seq differentially expressed genes revealed that, compared to the CTR group, genes related to the extracellular matrix were significantly up-regulated (P<0.05), while genes related to axon guidance were significantly down-regulated (P<0.05) in the SCC group on day 7 post-operation. On day 21, genes involved in immune regulation and the retinol signaling pathway were significantly activated in the SCC group (P<0.05). In contrast, in the HSCI group, genes associated with inflammation and immune response were significantly up-regulated (P<0.001), while genes related to neuronal differentiation and synapse formation were significantly down-regulated (P<0.001) on day 7. On day 21, genes related to cell-matrix junctions and N-methyl-D-aspartate receptors were significantly up-regulated (P<0.001) in the HSCI group. Furthermore, compared to the SCC group, the HSCI group exhibited different pathway enrichment characteristics in GO and KEGG analyses on days 7 and 21 post-injury. On day 7, genes involved in the NOD-like receptor signaling pathway and the complement and coagulation cascades were significantly up-regulated in the HSCI group (P<0.001). On day 21, genes related to the extracellular matrix-receptor interaction and the neuroactive ligand-receptor interaction pathways were significantly activated (P<0.001). Finally, real time fluorogenic quantitative PCR validation results were highly consistent with the RNA-Seq results, further confirming the differential expression trends of key genes between the SCC and HSCI groups.ConclusionThe SCC and HSCI injury models may drive distinct repair pathways: the preservation of some axons in the SCC model predisposes it toward tissue repair, whereas the HSCI model requires the coordination of more complex molecular networks to achieve a new equilibrium. This finding further deepens the understanding of the heterogeneous regulatory mechanisms underlying spinal cord injury.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 32 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.052
Exosomes Treat Ischemic Stroke by Regulation of Ferroptosis Through the NRF2/SLC7A11/GPX4 Pathway in Mice
XU Yingtao, WANG Mengmeng, LIN Ping, CHI Haitao, WANG Yi, and BAI Ying
ObjectiveA middle cerebral artery occlusion (MCAO) mouse model is established by electrocoagulation of the middle cerebral artery. The study examines the mechanism by which exosomes (EXO) derived from human amniotic mesenchymal stem cells (hAMSCs) improve ischemic stroke and regulate neural ferroptosis-related injury.MethodsThirty-two SPF-grade male C57BL/6J mice aged 6 - 8 weeks were randomly divided into four groups (n=8 per group): sham group (Sham), model group (MCAO), MCAO plus normal saline group (MCAO+NaCl), and MCAO plus exosome group (MCAO+EXO). The mouse MCAO model was established by electrocoagulation of the middle cerebral artery. Mice in the Sham group underwent exposure of the middle cerebral artery without electrocoagulation. Twenty-four hours before MCAO induction, mice in the MCAO+EXO group received a tail vein injection of 100 μL of exosomes derived from the culture supernatant of hAMSCs at a concentration of 9.5×1011 particles/mL. Mice in the MCAO+NaCl group were injected with an equal volume of normal saline via the tail vein. Twenty-four hours after model establishment, neurological deficits were evaluated using the Longa neurological deficit scoring system. Cerebral infarct volume was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Hematoxylin and eosin (HE) staining was performed to evaluate morphological changes of neurons in the ischemic brain regions. The contents of ferrous iron (Fe2+), malondialdehyde (MDA), total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) in the infarct core and peri-infarct regions were determined using microcolorimetric assays to evaluate differences among groups. The mRNA expression levels of ferroptosis-related factors, including nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in the infarct core and peri-infarct regions were measured by real-time quantitative PCR. Protein expression levels of NRF2, SLC7A11, and GPX4 in the infarct and peri-infarct regions of each group were analyzed by Western blotting.ResultsCompared with the MCAO group, the Longa neurological deficit score was significantly reduced in the MCAO+EXO group (P<0.01). Prominent cerebral infarction was observed in the MCAO group, whereas the infarct volume ratio was markedly decreased in the MCAO+EXO group compared with the MCAO group (P<0.001). Histopathological analysis revealed that mice in the MCAO group exhibited obvious neuronal damage, including cytoplasmic vacuolar degeneration, nuclear pyknosis and fragmentation, unclear nuclear structure, and disorganized neuronal arrangement, compared with the Sham group. In contrast, neurons in the MCAO+EXO group showed relatively preserved morphology, with intact cellular structures and large, regular nuclei located centrally within the cells. Biochemical analysis demonstrated that Fe2+ and MDA levels in the infarct core and peri-infarct regions were significantly increased in the MCAO group compared with the Sham group (P<0.001). These levels were significantly reduced in the MCAO+EXO group compared with the MCAO group (P<0.01). In addition, total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) levels were markedly decreased in the MCAO group relative to the Sham group (P<0.01). Compared with the MCAO group, the MCAO+EXO group exhibited significantly increased levels of total GSH and GSH (P<0.001), while no significant change was observed in GSSG levels (P>0.05). Furthermore, both mRNA and protein expression levels of nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) were significantly downregulated in the MCAO group compared with the Sham group (P<0.01, P<0.001). In contrast, both mRNA and protein expression levels of NRF2, SLC7A11, and GPX4 were significantly upregulated in the MCAO+EXO group compared with the MCAO group (P<0.05).ConclusionIn the mouse MCAO model, tail vein injection of exosomes derived from hAMSCs can improve motor function, reduce infarct area, protect neuronal cell morphology, and reduce the degree of nerve injury. Exosomes may exert a protective effect by activating the NRF2/SLC7A11/GPX4 pathway and reducing ferroptosis in neuronal cells of MCAO model mice.ObjectiveA middle cerebral artery occlusion (MCAO) mouse model is established by electrocoagulation of the middle cerebral artery. The study examines the mechanism by which exosomes (EXO) derived from human amniotic mesenchymal stem cells (hAMSCs) improve ischemic stroke and regulate neural ferroptosis-related injury.MethodsThirty-two SPF-grade male C57BL/6J mice aged 6 - 8 weeks were randomly divided into four groups (n=8 per group): sham group (Sham), model group (MCAO), MCAO plus normal saline group (MCAO+NaCl), and MCAO plus exosome group (MCAO+EXO). The mouse MCAO model was established by electrocoagulation of the middle cerebral artery. Mice in the Sham group underwent exposure of the middle cerebral artery without electrocoagulation. Twenty-four hours before MCAO induction, mice in the MCAO+EXO group received a tail vein injection of 100 μL of exosomes derived from the culture supernatant of hAMSCs at a concentration of 9.5×1011 particles/mL. Mice in the MCAO+NaCl group were injected with an equal volume of normal saline via the tail vein. Twenty-four hours after model establishment, neurological deficits were evaluated using the Longa neurological deficit scoring system. Cerebral infarct volume was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Hematoxylin and eosin (HE) staining was performed to evaluate morphological changes of neurons in the ischemic brain regions. The contents of ferrous iron (Fe2+), malondialdehyde (MDA), total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) in the infarct core and peri-infarct regions were determined using microcolorimetric assays to evaluate differences among groups. The mRNA expression levels of ferroptosis-related factors, including nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in the infarct core and peri-infarct regions were measured by real-time quantitative PCR. Protein expression levels of NRF2, SLC7A11, and GPX4 in the infarct and peri-infarct regions of each group were analyzed by Western blotting.ResultsCompared with the MCAO group, the Longa neurological deficit score was significantly reduced in the MCAO+EXO group (P<0.01). Prominent cerebral infarction was observed in the MCAO group, whereas the infarct volume ratio was markedly decreased in the MCAO+EXO group compared with the MCAO group (P<0.001). Histopathological analysis revealed that mice in the MCAO group exhibited obvious neuronal damage, including cytoplasmic vacuolar degeneration, nuclear pyknosis and fragmentation, unclear nuclear structure, and disorganized neuronal arrangement, compared with the Sham group. In contrast, neurons in the MCAO+EXO group showed relatively preserved morphology, with intact cellular structures and large, regular nuclei located centrally within the cells. Biochemical analysis demonstrated that Fe2+ and MDA levels in the infarct core and peri-infarct regions were significantly increased in the MCAO group compared with the Sham group (P<0.001). These levels were significantly reduced in the MCAO+EXO group compared with the MCAO group (P<0.01). In addition, total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) levels were markedly decreased in the MCAO group relative to the Sham group (P<0.01). Compared with the MCAO group, the MCAO+EXO group exhibited significantly increased levels of total GSH and GSH (P<0.001), while no significant change was observed in GSSG levels (P>0.05). Furthermore, both mRNA and protein expression levels of nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) were significantly downregulated in the MCAO group compared with the Sham group (P<0.01, P<0.001). In contrast, both mRNA and protein expression levels of NRF2, SLC7A11, and GPX4 were significantly upregulated in the MCAO+EXO group compared with the MCAO group (P<0.05).ConclusionIn the mouse MCAO model, tail vein injection of exosomes derived from hAMSCs can improve motor function, reduce infarct area, protect neuronal cell morphology, and reduce the degree of nerve injury. Exosomes may exert a protective effect by activating the NRF2/SLC7A11/GPX4 pathway and reducing ferroptosis in neuronal cells of MCAO model mice.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 20 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.034
Establishment and Multidimensional Pathological Evaluations of a Cigarette Smoke Exposure-Induced Chronic Obstructive Pulmonary Disease Mouse Model
HE Jiaqi, ZHOU Yuanyuan, NIE Yongqiang, WANG Zhaoxia, and XU Wangjie
ObjectiveTo establish a reliable chronic obstructive pulmonary disease (COPD) mouse model based on a self-developed multichannel automatic control system for long-term continuous cigarette smoke exposure in small animals using a novel continuous cigarette smoke exposure method, and to conduct phenotypic evaluation and analysis, thereby providing an animal experimental basis for investigating COPD pathogenesis and prevention strategies.MethodsTwenty male C57BL/6J mice aged 6 weeks were randomly and equally divided into a control group and a model group. The model group (n=10) underwent 6 h of continuous cigarette smoke exposure daily (6 cigarettes per day for 12 consecutive weeks), while the control group (n=10) received no intervention. Body weight was monitored biweekly. Post-exposure, in vivo micro-CT imaging was performed. After euthanasia, serum and bronchoalveolar lavage fluid (BALF) levels of interleukin-6 (IL-6) and tumor necrosis factor- (TNF-) were quantified by ELISA. Lung tissues underwent H&E and Masson's trichrome staining to observe changes in lung morphology and inflammatory cell infiltration, and the mean linear intercept (MLI) was calculated, thereby comprehensively evaluating the clinical features of COPD in the mouse model.ResultsCompared with the control group, the model group showed significantly reduced body weight (P<0.01) from the fourth week. Compared with the control group, IL-6 level in the serum and BALF of the model group increased by 27.2% and 140.0%, respectively (P<0.01). TNF- level in the serum and bronchoalveolar lavage fluid of the model group increased by 16.7% (P<0.01) and 19.3% (P<0.05), respectively. Histopathological examination revealed alveolar wall thinning, septal rupture, emphysematous bullae formation, reduced alveolar count, bronchial wall thickening with lumen narrowing, and inflammatory cell infiltration. MLI was significantly elevated (P<0.01). Masson's staining confirmed collagen deposition and bronchial remodeling. Micro-CT demonstrated localized high-density shadows exhibiting typical features of chronic bronchitis.ConclusionThe self-developed device enables long-term continuous smoke exposure, and the successfully established COPD mouse model exhibits pathological features highly consistent with clinical manifestations, offering an efficient and reliable tool for COPD research.ObjectiveTo establish a reliable chronic obstructive pulmonary disease (COPD) mouse model based on a self-developed multichannel automatic control system for long-term continuous cigarette smoke exposure in small animals using a novel continuous cigarette smoke exposure method, and to conduct phenotypic evaluation and analysis, thereby providing an animal experimental basis for investigating COPD pathogenesis and prevention strategies.MethodsTwenty male C57BL/6J mice aged 6 weeks were randomly and equally divided into a control group and a model group. The model group (n=10) underwent 6 h of continuous cigarette smoke exposure daily (6 cigarettes per day for 12 consecutive weeks), while the control group (n=10) received no intervention. Body weight was monitored biweekly. Post-exposure, in vivo micro-CT imaging was performed. After euthanasia, serum and bronchoalveolar lavage fluid (BALF) levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were quantified by ELISA. Lung tissues underwent H&E and Masson's trichrome staining to observe changes in lung morphology and inflammatory cell infiltration, and the mean linear intercept (MLI) was calculated, thereby comprehensively evaluating the clinical features of COPD in the mouse model.ResultsCompared with the control group, the model group showed significantly reduced body weight (P<0.01) from the fourth week. Compared with the control group, IL-6 level in the serum and BALF of the model group increased by 27.2% and 140.0%, respectively (P<0.01). TNF-α level in the serum and bronchoalveolar lavage fluid of the model group increased by 16.7% (P<0.01) and 19.3% (P<0.05), respectively. Histopathological examination revealed alveolar wall thinning, septal rupture, emphysematous bullae formation, reduced alveolar count, bronchial wall thickening with lumen narrowing, and inflammatory cell infiltration. MLI was significantly elevated (P<0.01). Masson's staining confirmed collagen deposition and bronchial remodeling. Micro-CT demonstrated localized high-density shadows exhibiting typical features of chronic bronchitis.ConclusionThe self-developed device enables long-term continuous smoke exposure, and the successfully established COPD mouse model exhibits pathological features highly consistent with clinical manifestations, offering an efficient and reliable tool for COPD research.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 11 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.076
Establishment and Preliminary Analysis of an AG6 Mouse Encephalopathy Model Induced by Vaccinia Virus Tiantan Strain Infection
YANG Lin, JIN Meng, WU Hanqing, LI Shun, and ZHOU Xiaohui
ObjectiveA mouse model of vaccinia virus Tiantan strain (VTT)-induced encephalopathy was developed using AG6 mice.MethodsVTT was amplified by infecting Vero cells at a multiplicity of infection (MOI) of 0.01, followed by concentration and titration. After 72 h of incubation, virus-containing cells were collected and subjected to concentration. The concentrated viral suspension was serially diluted (10-fold dilutions) and added to 6-well plates containing confluent Vero cell monolayers for plaque assay. The number of plaques formed in each well was counted, and the virus titer was calculated based on the dilution factor. Fourteen 5-6-week-old AG6 mice (half male and half female, housed separately by sex) were randomly divided into a control group (n=3, PBS), a low-dose group (n=6, 1×105 PFU), and a high-dose group (n=5, 5×105 PFU). The mice were anesthetized by isoflurane inhalation and then infected via intranasal instillation. The mental state of the mice in each group was observed daily, and the body weight and mortality were recorded. On day 13 post-infection, 2% Evans Blue (4 mL/kg body weight) was administered via tail vein injection to assess blood-brain barrier (BBB) disruption. Subsequently, brain tissue samples were collected for immunofluorescence analysis to evaluate the activation of astrocytes and microglia.ResultsThe titer of purified VTT was 1×107 PFU/mL. Compared with the control group, mice in the low-dose group showed no significant change in body weight, and no lethality was observed. In contrast, mice in the high-dose group exhibited significant weight loss starting on day 5 post-infection (P<0.05), accompanied by lethality. On day 13 post-infection, no Evans Blue extravasation was detected in the brain tissues of the low-dose group, while the olfactory bulb region of the high-dose group displayed distinct blue staining, indicating disruption of the BBB. Immunofluorescence analysis revealed no significant proliferation of astrocytes and microglia in the olfactory bulb region of the low-dose group on day 13 post-infection. In contrast, marked activation of glial cells was observable in the high-dose group.ConclusionAn animal model of VTT-induced encephalopathy in AG6 mice is successfully established, characterized by BBB disruption and reactive gliosis specifically localized to the olfactory bulb region, manifested as astrocytic and microglial proliferation.ObjectiveA mouse model of vaccinia virus Tiantan strain (VTT)-induced encephalopathy was developed using AG6 mice.MethodsVTT was amplified by infecting Vero cells at a multiplicity of infection (MOI) of 0.01, followed by concentration and titration. After 72 h of incubation, virus-containing cells were collected and subjected to concentration. The concentrated viral suspension was serially diluted (10-fold dilutions) and added to 6-well plates containing confluent Vero cell monolayers for plaque assay. The number of plaques formed in each well was counted, and the virus titer was calculated based on the dilution factor. Fourteen 5-6-week-old AG6 mice (half male and half female, housed separately by sex) were randomly divided into a control group (n=3, PBS), a low-dose group (n=6, 1×105 PFU), and a high-dose group (n=5, 5×105 PFU). The mice were anesthetized by isoflurane inhalation and then infected via intranasal instillation. The mental state of the mice in each group was observed daily, and the body weight and mortality were recorded. On day 13 post-infection, 2% Evans Blue (4 mL/kg body weight) was administered via tail vein injection to assess blood-brain barrier (BBB) disruption. Subsequently, brain tissue samples were collected for immunofluorescence analysis to evaluate the activation of astrocytes and microglia.ResultsThe titer of purified VTT was 1×107 PFU/mL. Compared with the control group, mice in the low-dose group showed no significant change in body weight, and no lethality was observed. In contrast, mice in the high-dose group exhibited significant weight loss starting on day 5 post-infection (P<0.05), accompanied by lethality. On day 13 post-infection, no Evans Blue extravasation was detected in the brain tissues of the low-dose group, while the olfactory bulb region of the high-dose group displayed distinct blue staining, indicating disruption of the BBB. Immunofluorescence analysis revealed no significant proliferation of astrocytes and microglia in the olfactory bulb region of the low-dose group on day 13 post-infection. In contrast, marked activation of glial cells was observable in the high-dose group.ConclusionAn animal model of VTT-induced encephalopathy in AG6 mice is successfully established, characterized by BBB disruption and reactive gliosis specifically localized to the olfactory bulb region, manifested as astrocytic and microglial proliferation.
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 3 (2026)
- DOI：10.12300/j.issn.1674-5817.2025.070
[in Chinese]
[in Chinese] and [in Chinese]
- May. 09, 2026
- Laboratory Animal and Comparative Medicine
- Vol. 46, Issue 1, 1 (2026)
- DOI：10.12300/j.issn.1674-5817.2026.027
Radiological pulp assessment of single-rooted teeth to estimate adult age: systematic review and meta-analysis
Angel Natania Hidayat, Elza Ibrahim Auerkari, Ferry Pergamus Gultom, Indah Melati Suciyanie, and and Pertti Auerkari
For disaster victim identification (DVI), the destruction of human remains can pose significant challenges in discerning useful identifying details. In such cases, dental age estimation may help to identify individuals, but tends to be less accurate for adults than for younger victims with less mature dentition. The Kvaal and Cameriere methods are used to estimate adult age by radiologically assessing pulp reduction resulting from the secondary dentin deposition in single-rooted teeth. This review aims to provide a meta-analysis of recent studies and their accuracy using Kvaal and Cameriere methods for adult age estimation from radiological pulp assessment. Searches were conducted in databases including Scielo, PubMed, EBSCO, Scopus, ScienceDirect, and Wiley Online Library. The searches were performed using the guidelines of Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). The search resulted in 1 241 studies of which 39 studies were eligible to be included. These studies comprised data from 12 distinct populations with a total of 7 510 samples. The Kvaal method underestimated (?2.95 years, 95%CI) and the Cameriere method overestimated (0.89 years, 95%CI) the assessed adult age in this meta-analysis. The Cameriere method presented lower mean difference and standard error of the estimation (SEE) compared to the Kvaal method. Overall, both methods can be used as adjunctive methods to estimate adult age through population-specific equations.Key PointsThis review presents an updated meta-analysis on the accuracy of two radiological pulp assessment methods, Kvaal and Cameriere, for dental age estimation.The Kvaal method underestimated and the Cameriere method overestimated the assessed age in the results of this meta-analysis.Type of radiograph, sample composition and size, methodology, population used, and observer differences are factors that affect the accuracy of these methods.For disaster victim identification (DVI), the destruction of human remains can pose significant challenges in discerning useful identifying details. In such cases, dental age estimation may help to identify individuals, but tends to be less accurate for adults than for younger victims with less mature dentition. The Kvaal and Cameriere methods are used to estimate adult age by radiologically assessing pulp reduction resulting from the secondary dentin deposition in single-rooted teeth. This review aims to provide a meta-analysis of recent studies and their accuracy using Kvaal and Cameriere methods for adult age estimation from radiological pulp assessment. Searches were conducted in databases including Scielo, PubMed, EBSCO, Scopus, ScienceDirect, and Wiley Online Library. The searches were performed using the guidelines of Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). The search resulted in 1 241 studies of which 39 studies were eligible to be included. These studies comprised data from 12 distinct populations with a total of 7 510 samples. The Kvaal method underestimated (?2.95 years, 95%CI) and the Cameriere method overestimated (0.89 years, 95%CI) the assessed adult age in this meta-analysis. The Cameriere method presented lower mean difference and standard error of the estimation (SEE) compared to the Kvaal method. Overall, both methods can be used as adjunctive methods to estimate adult age through population-specific equations.
Key Points
This review presents an updated meta-analysis on the accuracy of two radiological pulp assessment methods, Kvaal and Cameriere, for dental age estimation.
The Kvaal method underestimated and the Cameriere method overestimated the assessed age in the results of this meta-analysis.
Type of radiograph, sample composition and size, methodology, population used, and observer differences are factors that affect the accuracy of these methods.
- May. 06, 2026
- Forensic Sciences Research
- Vol. 11, Issue 1, tfsr20230179 (2026)
- DOI：10.3788/tfsr20230179
Study on the pharmacokinetics of 2-fluorodeschloroketamine and its metabolite in rats
Zhe Chen, Ye Liu, Huan Gao, Di Liang, Zhuoyi Wang, Chao Zhang, Jianguo Li, Zhiwen Wei, Dan Zhang, and Keming Yun
Currently, 2-fluorodeschloroketamine (2F-DCK), an analog of ketamine (KET), is considered one of the emerging new psychoactive substances (NPS). Its abuse is on the rise globally due to its structural and anesthetic similarities to KET. The pharmacological properties of KET have been extensively studied, but reports on the pharmacology of 2F-DCK and its metabolites are scarce. In the study, we focused on the pharmacokinetics of 2F-DCK and its phase I metabolite nor-2F-DCK in rodent model. Our aim was to analyse changes in their metabolic profiles over time and to construct a predictive model based on the concentration ratio of 2F-DCK to its metabolite for estimating the last 2F-DCK administration time. Eighteen healthy Sprague-Dawley (SD) male rats were randomly divided into three groups: high- and low-dose 2F-DCK groups and a control group (n = 6). Whole blood was collected from the inner canthal vein at different time intervals after oral gavage. The blood samples were extracted via liquid-liquid extraction and analysed by high-performance liquid chromatography?tandem mass spectrometry (HPLC-MS/MS) in a positive ion mode using multi-reaction monitoring. Mean blood concentration?time curves showed that 2F-DCK was undetectable at 8 h post-administration, whereas nor-2F-DCK was still detectable at 12 h (low-dose group), demonstrating that nor-2F-DCK could be used as a reliable biomarker for 2F-DCK identification. Pharmacokinetic parameters were calculated by a pharmacokinetic software DAS 3.2.8, employing a non-compartment model. The results showed the elimination half-lives of 2F-DCK are 0.20 h (low-dose group) and 1.07 h (high-dose group); while those of nor-2F-DCK were 1.51 h (low-dose group) and 5.01 h (high-dose group), respectively. The time of maximum concentration (Tmax) of 2F-DCK in low-dose group was same as that in high-dose group (0.25 h), and the Tmax of nor-2F-DCK in both groups was 1.00 h, which indicated that the Tmax was dose-independent. Significantly, the best predictive model was obtained from the relationship between the concentration ratio of 2F-DCK to nor-2F-DCK and the administration time of 2F-DCK, exhibiting considerable potential for application in clinical drug monitoring and forensic illegal drug identification.Currently, 2-fluorodeschloroketamine (2F-DCK), an analog of ketamine (KET), is considered one of the emerging new psychoactive substances (NPS). Its abuse is on the rise globally due to its structural and anesthetic similarities to KET. The pharmacological properties of KET have been extensively studied, but reports on the pharmacology of 2F-DCK and its metabolites are scarce. In the study, we focused on the pharmacokinetics of 2F-DCK and its phase I metabolite nor-2F-DCK in rodent model. Our aim was to analyse changes in their metabolic profiles over time and to construct a predictive model based on the concentration ratio of 2F-DCK to its metabolite for estimating the last 2F-DCK administration time. Eighteen healthy Sprague-Dawley (SD) male rats were randomly divided into three groups: high- and low-dose 2F-DCK groups and a control group (n = 6). Whole blood was collected from the inner canthal vein at different time intervals after oral gavage. The blood samples were extracted via liquid-liquid extraction and analysed by high-performance liquid chromatography?tandem mass spectrometry (HPLC-MS/MS) in a positive ion mode using multi-reaction monitoring. Mean blood concentration?time curves showed that 2F-DCK was undetectable at 8 h post-administration, whereas nor-2F-DCK was still detectable at 12 h (low-dose group), demonstrating that nor-2F-DCK could be used as a reliable biomarker for 2F-DCK identification. Pharmacokinetic parameters were calculated by a pharmacokinetic software DAS 3.2.8, employing a non-compartment model. The results showed the elimination half-lives of 2F-DCK are 0.20 h (low-dose group) and 1.07 h (high-dose group); while those of nor-2F-DCK were 1.51 h (low-dose group) and 5.01 h (high-dose group), respectively. The time of maximum concentration (Tmax) of 2F-DCK in low-dose group was same as that in high-dose group (0.25 h), and the Tmax of nor-2F-DCK in both groups was 1.00 h, which indicated that the Tmax was dose-independent. Significantly, the best predictive model was obtained from the relationship between the concentration ratio of 2F-DCK to nor-2F-DCK and the administration time of 2F-DCK, exhibiting considerable potential for application in clinical drug monitoring and forensic illegal drug identification.
- Apr. 20, 2026
- Forensic Sciences Research
- Vol. 11, Issue 1, tfsr20240235 (2026)
- DOI：10.3788/tfsr20240235
Removing watermark from diffusion models via Low-Rank Adaptation
Liting Zeng, Jinhua Zeng and Wei Lu
Diffusion models have emerged as powerful generative tools. Recently, numerous image watermarking techniques have been developed to ensure copyright protection and content traceability. These techniques also play a vital role in multimedia forensics, where digital watermarks are used to verify authenticity, detect tampering, and support legal attribution. Conventional watermark removal techniques operate at the image level, employing various post-processing strategies to degrade embedded watermarks. However, these methods often incur substantial computational costs, limiting their practical applicability. In this work, we propose a novel watermark removal approach targeting the diffusion model itself, specifically focusing on fine-tuning the Variational Autoencoder (VAE) decoder. Utilising Low-Rank Adaptation (LoRA), we efficiently optimise a subset of parameters to suppress watermark signals while preserving image fidelity. An adversarial classifier is incorporated to guide the training process, ensuring effective watermark removal. Furthermore, perceptual and structural similarity losses are integrated to mitigate degradation in image quality. Experimental results show that our approach effectively eliminates embedded watermarks, providing a robust and lightweight solution for watermark removal in diffusion-based generative models. This work also provides a technical perspective for analysing the reliability and resilience of watermark-based mechanisms in generative AI systems, offering insight into their potential limitations in forensic identification.Diffusion models have emerged as powerful generative tools. Recently, numerous image watermarking techniques have been developed to ensure copyright protection and content traceability. These techniques also play a vital role in multimedia forensics, where digital watermarks are used to verify authenticity, detect tampering, and support legal attribution. Conventional watermark removal techniques operate at the image level, employing various post-processing strategies to degrade embedded watermarks. However, these methods often incur substantial computational costs, limiting their practical applicability. In this work, we propose a novel watermark removal approach targeting the diffusion model itself, specifically focusing on fine-tuning the Variational Autoencoder (VAE) decoder. Utilising Low-Rank Adaptation (LoRA), we efficiently optimise a subset of parameters to suppress watermark signals while preserving image fidelity. An adversarial classifier is incorporated to guide the training process, ensuring effective watermark removal. Furthermore, perceptual and structural similarity losses are integrated to mitigate degradation in image quality. Experimental results show that our approach effectively eliminates embedded watermarks, providing a robust and lightweight solution for watermark removal in diffusion-based generative models. This work also provides a technical perspective for analysing the reliability and resilience of watermark-based mechanisms in generative AI systems, offering insight into their potential limitations in forensic identification.
- Apr. 14, 2026
- Forensic Sciences Research
- Vol. 11, Issue 1, tfsr20250147 (2026)
- DOI：10.3788/tfsr20250147
Estimating shooting distance from the inorganic components of GSRs by ICP-MS with Type 79 micro rifle
Xiang Li
A methodology to estimate shooting distance based on the analysis of inorganic components of gunshot residues (GSRs) was developed using inductively coupled plasma–mass spectrometry (ICP-MS). Test shots using a Type 79 micro rifle (250 mm barrel length and 7.62 × 25 mm cartridges) were made at distances between 10 and 90 cm. A series of mathematical models relating shooting distance to the content of elements Sn, Sb, Ba, and Pb in the GSRs was developed. These models were developed using data from a series of shots performed against a 45 × 45 cm white 100% polyester cloth. The target was sampled at five radial distances (in increasing 10 mm steps) and four perpendicular directions. A logarithmic correlation was observed between the shooting distance and the average content of elements Sn, Sb, Ba, and Pb at radial distances of 1.5 cm and 5.5 cm from the entrance hole, for shooting distances up to 90 cm.A methodology to estimate shooting distance based on the analysis of inorganic components of gunshot residues (GSRs) was developed using inductively coupled plasma–mass spectrometry (ICP-MS). Test shots using a Type 79 micro rifle (250 mm barrel length and 7.62 × 25 mm cartridges) were made at distances between 10 and 90 cm. A series of mathematical models relating shooting distance to the content of elements Sn, Sb, Ba, and Pb in the GSRs was developed. These models were developed using data from a series of shots performed against a 45 × 45 cm white 100% polyester cloth. The target was sampled at five radial distances (in increasing 10 mm steps) and four perpendicular directions. A logarithmic correlation was observed between the shooting distance and the average content of elements Sn, Sb, Ba, and Pb at radial distances of 1.5 cm and 5.5 cm from the entrance hole, for shooting distances up to 90 cm.
- Mar. 31, 2026
- Forensic Sciences Research
- Vol. 11, Issue 1, tfsr20240111 (2026)
- DOI：10.3788/tfsr20240111
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Special Issue on Debris produced in High Power Laser Interactions (2026)
Submission Open:23 April 2026; Submission Deadline: 31 August 2026
Special Issue on the 40th anniversary of National Laboratory on High Power Laser and Physics (2026)
Submission Open:15 April 2026; Submission Deadline: 15 August 2026
Special Issue on Forensic Document Examination (2024)
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