This cover image is based on the article “High‐throughput markerless pose estimation and home‐cage activity analysis of tree shrew using deep learning” reported by Yangzhen Wang, Feng Su, Rixu Cong, Mengna Liu, Kaichen Shan, Xiaying Li, Desheng Zhu, Yusheng Wei, Jiejie Dai, Chen Zhang, Yonglu Tian (https://doi.org/10.1002/ame2.12530). This study presents a deep learning approach that achieves markerless pose estimation and recognizes diverse natural behaviors in tree shrews. By capturing detailed body movements with high precision, this method offers an efficient and cost‐effective solution for studying tree shrews. The system enables high‐throughput, long‐term behavioral monitoring, making it a powerful tool for behavioral research. It offers new insights into their behavioral patterns and daily routines, deepening our understanding of their complex behaviors. [Image: see text]

Alzheimer's disease is the most prevalent chronic neurodegenerative disorder worldwide, with no sufficient cure. Ongoing research is focused on developing new therapies aimed at preventing or delaying the onset of symptoms, slowing disease progression, and improving cognitive and behavioral outcomes in individuals affected by Alzheimer's disease. Among the various pathological changes associated with this condition, blood-brain barrier (BBB) leakage plays a crucial role as it serves as a vital boundary for maintaining central nervous system (CNS) health. Preserving the integrity and functionality of the BBB is essential to protect the brain from amyloid-β accumulation, neuroinflammation, and neuronal degeneration. This review summarizes models of Alzheimer's disease characterized by BBB leakage over time. More importantly, we introduce Krüppel-like factor 4 (KLF4), a transcription factor involved in vascular systems, and discuss its relevance to Alzheimer's disease. By elucidating the functions of KLF4 within both vascular and CNSs, this review highlights its potential role in modulating BBB integrity in Alzheimer's pathology, which may contribute to therapeutic strategies for managing this debilitating condition.

BackgroundDry eye disease (DED) predominantly results from elevated tear film osmolarity, which can not only cause ocular inconvenience but may lead to visual impairments, severely compromising patient well‐being and exerting substantial economic burdens as well. Astaxanthin (AST), a member of the xanthophylls and recognized for its robust abilities to combat inflammation and oxidation, is a common dietary supplement. Nonetheless, the precise molecular pathways through which AST influences DED are still poorly understood.MethodsTherapeutic targets for AST were identified using data from the GeneCards, PharmMapper, and Swiss Target Prediction databases, and STITCH datasets. Similarly, targets for dry eye disease (DED) were delineated leveraging resources such as the Therapeutic Target Database (TTD), DisGeNET, GeneCards, and OMIM databases, and DrugBank datasets. Interactions among shared targets were charted and displayed using CytoScape 3.9.0. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses were conducted to elucidate the functions of pivotal targets within the protein–protein interaction network. Molecular interactions between AST and key targets were confirmed through molecular docking using AutoDock and PyMOL. Molecular dynamics simulations were performed using GROMACS 2022.3. Viability of human corneal epithelial cells (hCEC) was assessed across varying concentrations of AST. A mouse model of experimental DED was developed using 0.1% benzalkonium chloride (BAC), and the animals were administered 100 mg/kg/day of AST orally for 7 days. The efficacy of the treatments was assessed through a series of diagnostic tests to evaluate the condition of the ocular surface after the interventions. The levels of inflammation and oxidative stress were quantitatively assessed using methods such as reverse transcription‐polymerase chain reaction (RT‐PCR), Western blot, and immunofluorescence staining.ResultsNetwork pharmacology suggests that AST may alleviate DED by influencing oxidation–reduction signaling pathways and reducing oxidative stress provoked by BAC. In vivo experiments demonstrated an improved overall condition in AST‐administered mice in contrast to the control group. Immunofluorescence staining analyses indicated a decrease in Keap1 protein in the corneal tissues of AST‐treated mice and a significant increase in Nrf2 and HO‐1 protein. In vitro studies demonstrated that AST significantly enhanced cell viability and suppressed reactive oxygen species expression under hyperosmotic (HS) conditions, thereby protecting the human corneal epithelium.ConclusionAST is capable of shielding mice from BAC‐induced DED, decelerating the progression of DED, and mitigating oxidative stress damage under HS conditions in hCEC cells. The protective impact of AST on DED may operate through stimulating the Keap1‐Nrf2/HO‐1 signaling pathway. Our research findings indicate that AST may be a promising treatment for DED, offering new insights into DED treatment.

BackgroundThe composition of the intestinal flora and the resulting metabolites affect patients' sleep after surgery.MethodsWe intended to elucidate the mechanisms by which disordered intestinal flora modulate the pathophysiology of postoperative sleep disturbances in hosts. In this study, we explored the impacts of anesthesia, surgery, and postoperative sleep duration on the fecal microbiota and metabolites of individuals classified postprocedurally as poor sleepers (PS) and good sleepers (GS), as diagnosed by the bispectral index. We also performed fecal microbiota transplantation in pseudo‐germ‐free (PGF) rats and applied Western blotting, immunohistochemistry, and gut permeability analyses to identify the potential mechanism of its effect.ResultsResearch finding shows the PS group had significantly higher postoperative stool levels of the metabolites tryptophan and kynurenine than the GS group. PGF rats that received gut microbiota from PSs exhibited less rapid eye movement (REM) sleep than those that received GS microbiota (GS‐PGF: 11.4% ± 1.6%, PS‐PGF: 4.8% ± 2.0%, p < 0.001). Measurement of 5‐hydroxytryptophan (5‐HTP) levels in the stool, serum, and prefrontal cortex (PFC) indicated that altered 5‐HTP levels, including reduced levels in the PFC, caused sleep loss in PGF rats transplanted with PS gut flora. Through the brain–gut axis, the inactivity of tryptophan hydroxylase 1 (TPH1) and TPH2 in the colon and PFC, respectively, caused a loss of REM sleep in PGF rats and decreased the 5‐HTP level in the PFC.ConclusionsThese findings indicate that postoperative gut dysbiosis and defective 5‐HTP metabolism may cause postoperative sleep disturbances. Clinicians and sleep researchers may gain new insights from this study.

BackgroundQingyangshen (Cynanchum otophyllum C.K. Schneid) is a folk drug for treating depression and other mental disorders induced by social defeat stress. Neuroplasticity in the hippocampus is essential for the modulation of cognition and emotion, and its impairment may contribute to the development and progression of depression. Our previous studies have found that Qingyangshen glycosides (QYS) can improve depression‐like behavior in social failure mouse models, mainly through PGC‐1α/FNDC5/BDNF signaling pathways activation, but its effects and mechanisms on hippocampal neuroplasticity remain unknown.MethodsChronic social defeat stress (CSDS) was used to induce social defeat in mice. Morphological changes in the hippocampus were observed by H&E staining and Golgi staining. Immunofluorescence double staining was used to detect the expression of synaptophysin (SYN) and postsynaptic density protein‐95 (PSD‐95), while western blot was employed to evaluate PSD‐95, SYN, and doublecortin (DCX) proteins. The pathological processing of social defeat and the therapeutic effects of QYS on it was confirmed through behavioral assessment associated with morphologic observation.ResultsDuring the whole study, the sucrose preference indices and OFT activity time of CSDS mice were significantly decreased (p ≤ 0.05), and the tail suspension immobility time was significantly increased (p ≤ 0.05), suggesting that the mice had significant depressive symptoms. Treatment with QYS (25, 50, and 100 mg/kg) significantly alleviated depressive symptoms in CSDS mice, which was demonstrated by significantly (p ≤ 0.05 or p ≤ 0.01) reducing the duration of tail‐hanging immobility and increasing the tendency of sucrose preference indices and OFT activity time. QYS treatment also significantly increased the expression of DCX, PSD‐95, and SYN proteins, which play a crucial role in depression.ConclusionsQYS alleviated these symptoms by enhancing hippocampal neuroplasticity through upregulating the expression of synapse‐associated proteins (SAPs). The therapeutic mechanism of QYS may involve modulating the neuroplasticity of hippocampus neurons by altering the expression of SAPs.

Reliable animal models are crucial to drug development for focal segmental glomerulosclerosis (FSGS), a rare kidney disease. Variability in success rates in literature and significant ethical concerns with animal welfare necessitate further optimization of adriamycin (ADR)-induced FSGS model developed on BALB/c mice. High-performance liquid chromatography (HPLC) was used to assess ADR stability in water and upon light exposure. To identify the optimal ADR level, single intravenous ADR injections with dosing levels from 10 to 17 mg/kg body weight were administered to BALB/c mice to induce FSGS-like pathology. Body weight and proteinuria of FSGS mice were monitored and analyzed for FSGS model-associated morbidity. Animals were euthanized for hematological and kidney histological assessments 8 weeks post induction. To identify the suitable experiment time frame of the ADR-induced FSGS mouse model, a longitudinal study was performed, with an 11-week continuous monitoring of the symptoms. ADR was found to be unstable in aqueous media and light sensitive. A dosing level of 10.5 mg/kg of ADR was optimal for consistent FSGS mouse model induction on BALB/c strain, characterized by minimal mortality and sustained FSGS-like symptoms. Findings from the longitudinal study suggest that 6 weeks post ADR induction may represent the peak of FSGS pathology severity in this mouse model. This time frame may be used for FSGS drug development projects. Based on the outcome from this study, we identified the optimal ADR dosing level and model testing duration. A standard operating procedure (SOP) for the ADR-induced FSGS mouse model was established to facilitate FSGS basic research and drug development.

Acquired immune deficiency syndrome (AIDS) is the name used to describe several potentially life‐threatening infections and disorders that happen when HIV has severely compromised the immune system. The primary effect of HIV is to decrease host immunity, exposing the host to external pathogens. The development of pharmaceutical drugs that directly cure the infection is crucial because of the current wide‐ranging epidemic of HIV. Most therapeutic anti‐HIV drugs are nucleosides. However, their high toxicity and potential for drug resistance restrict their use. Many of the most effective clinical drugs used to inhibit HIV, the activation of latent HIV, and AIDS have been obtained from natural sources. This review focuses on potential natural medicinal products for treating and managing HIV and AIDS. Notwithstanding, further clinical research studies are needed to understand the subject and its dynamics.

BackgroundAortic atherosclerosis increases the risk of embolic events under extracorporeal circulation (ECC). To evaluate the hemodynamic impact of ECC on atheromatous plaques, an atherosclerosis animal model, which is also eligible for ECC, is required.MethodsTwenty‐nine New Zealand White rabbits received a pro‐atherosclerotic diet (group diet, n = 10), a pro‐atherosclerotic diet and additional intraaortic balloon insufflation injury (group BI, n = 9), or served as controls (n = 10). After 3 or 6 months, aortic explants were analyzed by (immuno‐)histology and RT‐PCR.ResultsBlood serum analyses revealed increased cholesterol‐levels in groups diet and BI compared to controls (3 months: p = 0.03 each, 6 months: p < 0.0001 each). Aortic inflammatory infiltration was significantly enhanced in groups diet (CD3 at 3 months: p < 0.0001, 6 months: p = 0.02; CD68 at 3 months: p = 0.01) and BI (CD3 at 3 months: p < 0.0001, 6 months: p = 0.03; CD68 at 3 months: p = 0.04, 6 months: p = 0.02). Increased intima hyperplasia occurred in both groups (p < 0.0001 each). Macroscopic analyses after 3 and 6 months showed ubiquitous lumen‐narrowing aortic plaques. Calcification of the intima and media was increased in groups diet (intima: p < 0.0001 at 3 and 6 months; media at 3 months: p < 0.0001, 6 months: p = 0.01) and BI (intima: p < 0.0001 at 3 and 6 months; media at 3 months: p < 0.0001, 6 months: p = 0.02). Extensive lipid accumulation was found in the intima in both treatment groups (p < 0.0001 each).ConclusionsA rabbit model with high aortic calcific plaque burden—diet‐induced with no implicit need of an additional intimal injury by an intraaortic balloon insufflation due to comparable outcome—exhibiting multiple pathophysiological aspects of human atherosclerosis has been designed and thoroughly characterized. It is suitable for use in future studies on the interaction between atherosclerotic plaques and the arterial blood flow under ECC.

The high morbidity and mortality of colorectal cancer (CRC) is a major challenge in clinical practice. Although a series of alternative research models of CRC have been developed, appropriate orthotopic animal models that reproduce the specific clinical response as well as pathophysiological immune features of CRC are still lacking. In the current study, we constructed a CRC orthotopic xenograft model by implanting the tumor tubes at the colorectum of mice and monitored the model development using bioluminescence imaging. This model successfully recapitulates the clinical chemotherapy efficacy, including reduced total flux, tumor weight, and the expression of Ki67 after treatment of the first‐line chemotherapy regime of CRC (FOLFOX: oxaliplatin and 5‐fluorouracil/calcium folinate). The model also reproduced the immunosuppressive effect of FOLFOX, indicated by decreased infiltration of macrophages and increased Treg cells in tumor. Additionally, the orthotopic xenograft approach may be applied in immunodeficient NCG/NSG mice for constructing patient‐derived xenografts, and being used in clinical precision medicine and drug evaluation. We believe the current model is a successful surgical orthotopic xenograft approach for cancer research and deserves to be popularized, which will provide a convenient and efficient platform for in‐depth mechanism exploration of CRC and preclinical drug evaluation.