Ferroptosis is a regulated form of cell death, with its core mechanism being intracellular iron overload-induced lipid peroxidation, leading to cellular dysfunction and mitochondrial structural abnormalities. Ferroptosis is closely related to various diseases including neurodegenerative disorders, tumors, and ischemia-reperfusion organ damage, and has become a potential therapeutic target. Iron is essential for life but can also cause cell death. Despite continuous progress in iron-related biomedical research, many questions remain unanswered. Advances in high-throughput technologies, genomics and proteomics are expected to reveal the cellular iron regulatory mechanism and open up new therapeutic approaches for ferroptosis-related diseases. This article reviews the research progress on iron in terms of its biology, metabolism, regulation, and related diseases, aiming to provide clues and references for developing new ferroptosis-targeted therapeutic strategies and facilitating more in-depth molecular studies from multiple perspectives.
Humans ; Ferroptosis/physiology* ; Iron/metabolism* ; Animals ; Neoplasms/metabolism* ; Neurodegenerative Diseases/metabolism*

Animal experiments are an essential component of life sciences and medical research. However, the external validity and reliability of individual animal studies are frequently challenged by inherent limitations such as small sample sizes, high design heterogeneity, and poor reproducibility, which impede the effective translation of research findings into clinical practice. Systematic reviews and meta-analysis represent a key methodology for integrating existing evidence and enhancing the robustness of conclusions. Currently, however, the application of systematic reviews and meta-analysis in the field of animal experiments lacks standardized guidelines for their conduct and reporting, resulting in inconsistent quality and, to some extent, diminishing their evidence value. To address this issue, this paper aims to systematically delineate the reporting process for systematic reviews and meta-analysis of animal experiments and to propose a set of standardized recommendations that are both scientific and practical. The article's scope encompasses the entire process, from the preliminary preparatory phase [including formulating the population， intervention， comparison and outcome （PICO） question, assessing feasibility, and protocol pre-registration] to the key writing points for each section of the main report. In the core methods section, the paper elaborates on how to implement literature searches, establish eligibility criteria, perform data extraction, and assess the risk of bias, based on the Preferred Reporting Items for Systematic Reviews and Meta-analysis （PRISMA) statement, in conjunction with relevant guidelines and tools such as Animal Research： Reporting of in Vivo Experiments （ARRIVE) and a risk of bias assessment tool developed by the Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE). For the presentation of results, strategies are proposed for clear and transparent display using flow diagrams and tables of characteristics. The discussion section places particular emphasis on how to scientifically interpret pooled effects, thoroughly analyze sources of heterogeneity, evaluate the impact of publication bias, and cautiously discuss the validity and limitations of extrapolating findings from animal studies to clinical settings. Furthermore, this paper recommends adopting the Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology to comprehensively grade the quality of evidence. Through a modular analysis of the entire reporting process, this paper aims to provide researchers in the field with a clear and practical guide, thereby promoting the standardized development of systematic reviews and meta-analysis of animal experiments and enhancing their application value in scientific decision-making and translational medicine.

Objective:To explore the expression and ultrastructure distribution of cofilin and its potential mecha-nisms in postsynaptic mitochondrial anchoring and synaptic plasticity in the pre-B?tzinger complex(pre-B?tC)in rats.Methods:Using neurokinin 1 receptor(NK1R)as a morphological marker of the pre-B?tC,we examined the expres-sion and ultrastructure distribution of cofilin in pre-B?tC neurons of normoxic(NOR)and acute intermittent hypoxic(AIH)rats by combining Western blot,RT-qPCR,immunofluorescence,and immunoelectron microscopy.Results:Cofilin was expressed in the nuclei,somata and dendrites of NK1R-positive neurons in the pre-B?tC and its expression decreased after AIH intervention in this region detected.The results of immunoelectron microscopy showed that cofilin was expressed in the dendritic spines and postsynaptic filamentous or flocculent structures of pre-B?tC neurons.A total of 317 synapses were detected in pre-B?tC(NOR:122;AIH:195).In the NOR group,65.6％of the postsynaptic had cofilin expression,while in the AIH group,the proportion decreased to 47.2％.Combining with the role of cofilin in the regulation of actin severing,it is suggested that the severing effect of cofilin on postsynaptic actin is weakened af-ter AIH intervention,which may further enhance the postsynaptic mitochondrial anchoring.Conclusion:Cofilin,an ac-tin binding protein,plays a crucial role in regulating shearing and depolymerization of actin.Decreased cofilin expres-sion induced by AIH suggests an enhanced actin polymerization,which may be involved in the postsynaptic anchoring of mitochondria and the regulation of synaptic plasticity in the pre-B?tC.

Objective:To translate the quality of life tool for patients with aplastic anaemia and paroxysmal nocturnal haemoglobinuria (QLQ-AA/PNH) into Chinese, and to test its reliability and validity.Methods:According to the scale translation principle, the Chinese version of QLQ-AA/PNH was formed through translation, back translation and cross-cultural adaptation. A cross-sectional survey method was used to conveniently select 58 patients with aplastic anemia who were treated in the hematology department of the Second Affiliated Hospital of Dalian Medical University from January 2018 to September 2023 for investigation, and to evaluate the reliability and validity of the scale.Results:The Chinese version of QLQ-AA/PNH retains 36 items, and 5 common factors (psychological status dimension, life burden dimension, physical condition dimension, illness anxiety dimension and other symptom dimension) were extracted through exploratory factor analysis. The cumulative variance contribution rate reached 71.33%, and the factor load of each entry was greater than 0.5 on corresponding common factors. The Cronbach α coefficient of the scale as a whole was 0.971, the broken half reliability coefficient was 0.985, the Cronbach α coefficient of each common factor was 0.637 to 0.954, and the broken half reliability coefficient was 0.637 to 0.930. Conclusions:The Chinese version of QLQ-AA/PNH has been proved to be valid and reliable. It is a valuable tool for evaluating the quality of life among patients with aplastic anaemia.

Objective:To explore the expression and ultrastructure distribution of cofilin and its potential mecha-nisms in postsynaptic mitochondrial anchoring and synaptic plasticity in the pre-B?tzinger complex(pre-B?tC)in rats.Methods:Using neurokinin 1 receptor(NK1R)as a morphological marker of the pre-B?tC,we examined the expres-sion and ultrastructure distribution of cofilin in pre-B?tC neurons of normoxic(NOR)and acute intermittent hypoxic(AIH)rats by combining Western blot,RT-qPCR,immunofluorescence,and immunoelectron microscopy.Results:Cofilin was expressed in the nuclei,somata and dendrites of NK1R-positive neurons in the pre-B?tC and its expression decreased after AIH intervention in this region detected.The results of immunoelectron microscopy showed that cofilin was expressed in the dendritic spines and postsynaptic filamentous or flocculent structures of pre-B?tC neurons.A total of 317 synapses were detected in pre-B?tC(NOR:122;AIH:195).In the NOR group,65.6％of the postsynaptic had cofilin expression,while in the AIH group,the proportion decreased to 47.2％.Combining with the role of cofilin in the regulation of actin severing,it is suggested that the severing effect of cofilin on postsynaptic actin is weakened af-ter AIH intervention,which may further enhance the postsynaptic mitochondrial anchoring.Conclusion:Cofilin,an ac-tin binding protein,plays a crucial role in regulating shearing and depolymerization of actin.Decreased cofilin expres-sion induced by AIH suggests an enhanced actin polymerization,which may be involved in the postsynaptic anchoring of mitochondria and the regulation of synaptic plasticity in the pre-B?tC.

Objective:To translate the quality of life tool for patients with aplastic anaemia and paroxysmal nocturnal haemoglobinuria (QLQ-AA/PNH) into Chinese, and to test its reliability and validity.Methods:According to the scale translation principle, the Chinese version of QLQ-AA/PNH was formed through translation, back translation and cross-cultural adaptation. A cross-sectional survey method was used to conveniently select 58 patients with aplastic anemia who were treated in the hematology department of the Second Affiliated Hospital of Dalian Medical University from January 2018 to September 2023 for investigation, and to evaluate the reliability and validity of the scale.Results:The Chinese version of QLQ-AA/PNH retains 36 items, and 5 common factors (psychological status dimension, life burden dimension, physical condition dimension, illness anxiety dimension and other symptom dimension) were extracted through exploratory factor analysis. The cumulative variance contribution rate reached 71.33%, and the factor load of each entry was greater than 0.5 on corresponding common factors. The Cronbach α coefficient of the scale as a whole was 0.971, the broken half reliability coefficient was 0.985, the Cronbach α coefficient of each common factor was 0.637 to 0.954, and the broken half reliability coefficient was 0.637 to 0.930. Conclusions:The Chinese version of QLQ-AA/PNH has been proved to be valid and reliable. It is a valuable tool for evaluating the quality of life among patients with aplastic anaemia.

Data analysis models may assist the transmission of traditional Chinese medicine （TCM） experience and clinical diagnosis and treatment， and the possibility of constructing a “data-knowledge” dual-drive model was explored by taking gastric precancerous state as an example. Data-driven is to make clinical decisions around data analysis， and its syndrome-differentiation decision-making research relies on hidden structural models and partially observable Markov decision-making processes to identify the etiology of diseases， syndrome elements， evolution of pathogenesis， and syndrome differentiation protocols； knowledge-driven is to make use of data and information to promote decision-making and action processes， and its syndrome-differentiation decision-making research relies on convolutional neural networks to improve the accuracy of local disease identification and syndrome differentiation. The “data-knowledge” dual-driven model can make up for the shortcomings of single-drive numerical simulation accuracy， and achieve a balance between local disease identification and macroscopic syndrome differentiation. On the basis of previous research， we explored the construction method of diagnostic assisted decision-making platform for gastric precancerous state， and believed that the diagnostic and decision-making ability of doctors can be extended through the assistance of machines and algorithms. Meanwhile， the related research methods were integrated and the core features of gastric precancerous state based on TCM syndrome differentiation and endoscopic pathology diagnosis and prediction were obtained， and the elements of endoscopic pathology recognition based on TCM syndrome differentiation were explored， so as to provide ideas for the in-depth research and innovative application of cutting-edge data analysis technology in the field of intelligent TCM syndrome differentiation.

Objective:To explore the protective effect of miR-346 up-regulated by crocin on myocardial ischemia reperfusion injury in rats.Methods:The rat model of myocardial ischemia reperfusion was constructed by open-chest ligation of the left anterior descending coronary artery followed by reperfusion. Fifty male SD rats were randomly divided into the sham operation group, model group, crocin group, miR-346 negative control (miR-NC) group, and miR-346 inhibitor group with 10 rats in each group. Rats in the sham operation group were only received thoracotomy without ligation. Rats in the miR-NC group and miR-346 inhibitor group were injected with miR-NC or miR-346 inhibitor through the tail vein 48 h prior to ligation of the left anterior descending coronary artery ligation. The crocetin reagent for gastric lavage was prepared by dissolving 8 mg of crocetin in 100 ml of physiological saline. Rats in the crocetin group, miR-NC group, and miR-346 inhibitor group were gavaged with crocetin reagent at 80 mg/kg. Rats in the sham operation group and model group were gavaged with saline at 5 ml/kg. The crocetin reagent and saline were gavaged once a day for 15 days. Serum levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) were detected by enzyme-linked immunosorbent assay (ELISA). The miR-346 expression level was detected by real-time fluorescence quantitative PCR (qRT-PCR). The pathological changes in cardiac muscle tissues were detected by HE staining. Cardiomyocyte apoptosis rate was detected by TUNEL staining. The expression levels of apoptosis-related proteins were detected by Western Blot.Results:Compared with the sham operation group, the serum levels of CK-MB and LDH, pathological scores, cardiomyocyte apoptosis rate and expression of cleaved cysteinyl aspartate specific proteinase-3 (cleaved Caspase-3) and B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax) were increased in the model group (all P < 0.05), and the miR-346 expression level and Bcl-2 levels in myocardial tissues were decreased (all P < 0.05). Compared with the model group, serum CK-MB and LDH levels, pathological scores, cardiomyocyte apoptosis rate, and cleaved Caspase-3 and Bax expression levels were decreased in the crocin group (all P < 0.05), and miR-346 expression level and Bcl-2 levels in the myocardial tissues were increased (all P < 0.05). Compared with the miR-NC group, the serum levels of CK-MB, LDH, pathological scores, cardiomyocyte apoptosis rate, and cleaved Caspase-3 and Bax expression levels were decreased in the miR-346 inhibitor group (all P < 0.05), and miR-346 expression level and Bcl-2 level were increased in myocardial tissues (all P < 0.05). Conclusions:Crocin can reduce myocardial tissue injury and attenuate cardiomyocyte apoptosis in myocardial ischemia-reperfusion rats by up-regulating miR-346.

This study performed to determine whether OX40L overexpressed by recombinant rabies virus(LBNSE-OX40L)can enhance the innate immune response through activation of dendritic cells and thus activate early antibody production.Bone marrow dendritic cells(BMDCs)were extracted from the femur of Balb/c mice and cultured for 6 days,and the cultured BMDCs were infected with the parental virus LBNSE and the recombinant virus LBNSE-OX40L with the multiplicity of infections(MOI)=1.The effect of each virus on the maturation of BMDCs was analyzed by flow cytometry;ELISA was used to detect the expression of innate immunity-related cytokines such as interferon-α(IFN-α)and interleukin-12p40(IL-12p40)in the supernatants of the infected BMDCs.For in vivo study,Balb/c female mice were injected intramuscularly with 106 FFU of parental virus LBNSE and recombinant virus LBNSE-OX40L in both hind limbs,and the inguinal lymph nodes of mice were collected on day 6 after immunization,and the proportion of mature DCs was detected by flow cytometry.The serum was collected on day 6 after immunization,and the content of virus-neutralizing antibody(VNA)was detected by antiviral neutralizing antibody titration.Mouse serum was collected on day 6 after immunization,and virus neutralizing antibody content was measured by titration of antiviral neutralizing antibody,while IgG antibody in mouse serum was detected by ELISA.IgM antibody subclasses were detected by ELISA on days 2,4,and 6 after immunization.Compared with the parental virus LBNSE,the recombinant virus LBNSE-OX40L was able to activate more BMDCs in vitro and produce significantly higher levels of IFN-α and IL-12p40.Furthermore,the recombinant virus LBNSE-OX40L stimulated the maturation and differentiation of the DCs in vivo,which led to the rapid production of high levels of VNA and RABV-specific IgG and IgM antibodies.Taken together,LBNSE-OX40L activates dendritic cells to promote the body's innate immune response,and in turn enhances early antibody production,thus can be an early effective rabies vaccine candidate.

Objective:To investigate the feasibility of simultaneous arteriovenous enhancement of neck CT with two-stage injection of contrast agent and its effect on image quality and radiation dose.Methods:A total of 30 patients undergoing neck CT enhancement scan due to space-occupying lesions in Beijing Tongren Hospital, Capital Medical University from February to April 2022 were prospectively included as the experimental group. The neck CT enhancement scan was performed with two-stage injection of contrast agent and arteriovenous simultaneous enhancement. The dosage of contrast agent was calculated according to the patient′s body weight, and the method of two-stage injection was adopted. The dosage of contrast agent in the first stage was 0.7 ml/kg, with normal saline in the middle stage, and the second stage (began at 35 s) was 0.3 ml/kg. A total of 30 patients with gender and age matching with the experimental group from December 2021 to January 2022 were retrospectively collected as the control group. The control group was treated with the traditional arterial phase and venous phase scanning method with the dosage of 1.0 ml/kg contrast agent. The arterial phase was scanned at the 30 s and the venous phase was scanned at the 60 s. The CT values of bilateral carotid arteries and jugular veins in the experimental group were measured, the CT values of bilateral carotid arteries in the arterial phase were measured in the control group, and the CT values of bilateral carotid arteries and jugular veins in the venous phase were measured. Carotid artery enhancement score was performed for images of experimental group and control group in arterial and venous phase, and jugular vein and lesion enhancement score was performed for images of experimental group and control group in venous phase. The effective dose was calculated for both groups. The difference of carotid artery CT values between images was compared by one-way analysis of variance, and LSD method was used for pairwise comparison. The CT values of jugular vein were compared using independent sample t test. Kruskal-Wallis test was used to compare carotid artery enhancement scores, and Nemenyi method was used for pairwise comparison. Jugular vein and lesion enhancement scores and effective dose were compared by Mann-Whitney U test. Results:The CT value of carotid artery of experimental group [left (276±24) HU, right (273±25) HU] was lower than that of control group in arterial phase [left (329±33) HU, right (327±32) HU], and higher than that in the venous phase [left (147±15) HU, right (148±16) HU]. All the differences were statistically significant ( P<0.001). The CT value of jugular vein of experimental group [left (206±18) HU, right (203±19)] was higher than that of control group in the venous phase [left (154±15) HU, right (151±15)], the difference was statistically significant ( t=11.88, 11.76, both P<0.001). There was no significant difference in carotid artery enhancement score between experimental group and control group in arterial phase ( P=0.624), but the carotid artery enhancement score of the experimental group was higher than that of the control group in the venous phase, and the difference was statistically significant ( P<0.001). The scores of jugular vein and lesion enhancement in experimental group were higher than those of control group in venous phase, and the difference was statistically significant ( Z=5.01, P<0.001). The effective dose of the experimental group [2.41(2.04, 2.72) mSv] was decreased by 52.2% compared with the control group [5.04(4.18, 5.44) mSv], and the difference was statistically significant ( Z=-6.24, P<0.001). Conclusions:The neck CT enhanced scan with two-stage injection of contrast agent and arteriovenous simultaneous enhancement method can obtain comprehensive images of arterial and venous phases, and realize simultaneous enhancement of carotid artery, jugular vein and lesions, and reduce radiation dose.