ObjectiveThe malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells (RBCs), facilitating their intracellular survival and pathogenicity. Skeleton-binding protein 1 (SBP1) is a conserved exported protein across Plasmodium species. In Plasmodium falciparum, SBP1 has been reported to interact with erythrocyte membrane skeleton proteins 4.1R and spectrin, while its contribution to erythrocyte remodeling and parasite virulence in Plasmodium berghei (Pb) remains unclear. This study aims to determine whether PbSBP1 associates with the host cytoskeletal protein 4.1R and to investigate its role in the remodeling of host RBCs and the pathogenicity of Plasmodium berghei. MethodsIn Plasmodium berghei, the relationship between PbSBP1 and the erythrocyte cytoskeletal protein 4.1R was examined using co-immunoprecipitation. A Pbsbp1 gene knockout mutant of Plasmodium berghei (Pbsbp1∆) was generated based on the principle of double crossover homologous recombination. The deformability of erythrocytes infected with Pbsbp1∆ parasites was assessed using microfluidic methods. Microchannels with an array of cylindrical pillars were used to detect modifications in infected RBC deformability. The infected RBCs were squashed between the rows and recovered between the columns and the transit velocity (μm/s) of infected RBCs travelling through the microchannel was recorded. The component of the erythrocyte membrane skeleton junctional complex, tropomodulin (TMOD), was fluorescently labeled, and the cytoskeletal network of infected erythrocytes was imaged using super-resolution stochastic optical reconstruction microscopy (STORM) to analyze ultrastructural changes in the cytoskeleton of wild-type (WT) and Pbsbp1∆-infected erythrocytes. Actin-based junctional complexes were displayed as individual clusters by the labeled TMOD in the STORM images, and the cluster densities and distances between adjacent clusters of infected RBCs were calculated. Additionally, rodent malaria models (BALB/c mice) and experimental cerebral malaria models (C57BL/6 mice) were employed to monitor the growth of Pbsbp1∆ and WT parasites during the intraerythrocytic stage and their capacity to induce cerebral malaria in mice. ResultsPbSBP1 may participate in the remodeling of infected erythrocytes through direct or indirect interaction with the erythrocyte cytoskeletal protein 4.1R. Microfluidic assays revealed that the deformability of erythrocytes infected with Pbsbp1∆ parasites was significantly enhanced compared to those infected with WT parasites. STORM imaging further demonstrated that the ultrastructure of the erythrocyte cytoskeleton in Pbsbp1∆-infected cells was altered relative to that in WT-infected erythrocytes. The distances between nearest neighbors of clusters had a tendency to increase while the cluster densities were decreased in Pbsbp1∆-infected RBCs compared to WT-infected RBCs. Subsequent phenotypic analysis indicated that the growth rate of Pbsbp1∆ parasites during the intraerythrocytic stage was significantly slower than that of WT parasites, and their ability to induce cerebral malaria in mice was also attenuated. These findings suggest that PbSBP1 is involved in the remodeling of the erythrocyte membrane skeleton, likely through its direct or indirect interaction with protein 4.1R, thereby regulating the deformability of infected erythrocytes and influencing the pathogenicity of the blood-stage parasites. ConclusionThis study establishes a role for PbSBP1 in host erythrocyte remodeling and parasite virulence, providing new research strategies for the prevention and treatment of malaria.

Subacute cholecystitis complicated by gallbladder perforation often manifests as inflammatory lesions around the gallbladder， and it is extremely rare for this condition to cause massive hemorrhage into the omental bursa with secondary infection and result in massive hemorrhagic abscess. This article reports a case of a female patient， aged 68 years， who experienced massive hemorrhagic abscess in the omental bursa due to localized perforation and bleeding caused by a stone impacted in the gallbladder neck. The patient had a history of gallstones for 30 years and was admitted to Tianjin Nankai Hospital， Tianjin Medical University， after tertiary referral due to aggravated abdominal pain， chills， and fever. After comprehensive assessment by a multidisciplinary team， the patient underwent emergency laparoscopic cholecystectomy and clearance of the hemorrhagic abscess in the omental bursa. The patient recovered well and was discharged on day 10 after surgery， with good conditions during follow-up.

Diabetic cataract is a prevalent and severe complication of diabetes mellitus. Its pathogenesis is complex and driven by multiple metabolic disorders. Conventional research methods are insufficient to fully clarify its complex pathophysiological processes. As a high-throughput and unbiased systems biology approach, metabolomics allows comprehensive profiling of small-molecule metabolites in biological systems, serving as a robust tool for exploring the metabolic signatures underlying this disease. This review systematically summarizes recent progress in the application of metabolomics to diabetic cataract research, with a focus on technological innovations in sample pretreatment, data acquisition, and bioinformatics analysis. This article elaborates on how these advances contribute to the identification of disease-specific biomarkers, the clarification of intrinsic molecular mechanisms, and the discovery of potential therapeutic targets, ultimately advancing the translational application of metabolomics in the early clinical diagnosis, prognostic assessment, and development of innovative therapeutic strategies for diabetic cataract.

Purpose: Leptin interacts not only with leptin receptor (LEPR) but also engages with other receptors. While the pro-oncogenic effects of the adrenergic receptor β2 (ADRB2) are well-established, the role of leptin in activating ADRB2 in triple-negative breast cancer (TNBC) remains unclear. Materials and Methods: The pro-carcinogenic effects of LEPR were investigated using murine TNBC cell lines, 4T1 and EMT6, and a tumor-bearing mouse model. Expression levels of LEPR, NADPH oxidase 4 (NOX4), and ADRB2 in TNBC cells and tumor tissues were analyzed via western blot and quantitative real-time polymerase chain reaction. Changes in reactive oxygen species (ROS) levels were assessed using flow cytometry and MitoSox staining, while immunofluorescence double-staining confirmed the co-localization of LEPR and ADRB2. Results: LEPR activation promoted NOX4-derived ROS and mitochondrial ROS production, facilitating TNBC cell proliferation and migration, effects which were mitigated by the LEPR inhibitor Allo-aca. Co-expression of LEPR and ADRB2 was observed on cell membranes, and bioinformatics data revealed a positive correlation between the two receptors. Leptin activated both LEPR and ADRB2, enhancing intracellular ROS generation and promoting tumor progression, which was effectively countered by a specific ADRB2 inhibitor ICI118551. In vivo, leptin injection accelerated tumor growth and lung metastases without affecting appetite, while treatments with Allo-aca or ICI118551 mitigated these effects. Conclusion This study demonstrates that leptin stimulates the growth and metastasis of TNBC through the activation of both LEPR and ADRB2, resulting in increased ROS production. These findings highlight LEPR and ADRB2 as potential biomarkers and therapeutic targets in TNBC.

Purpose: Leptin interacts not only with leptin receptor (LEPR) but also engages with other receptors. While the pro-oncogenic effects of the adrenergic receptor β2 (ADRB2) are well-established, the role of leptin in activating ADRB2 in triple-negative breast cancer (TNBC) remains unclear. Materials and Methods: The pro-carcinogenic effects of LEPR were investigated using murine TNBC cell lines, 4T1 and EMT6, and a tumor-bearing mouse model. Expression levels of LEPR, NADPH oxidase 4 (NOX4), and ADRB2 in TNBC cells and tumor tissues were analyzed via western blot and quantitative real-time polymerase chain reaction. Changes in reactive oxygen species (ROS) levels were assessed using flow cytometry and MitoSox staining, while immunofluorescence double-staining confirmed the co-localization of LEPR and ADRB2. Results: LEPR activation promoted NOX4-derived ROS and mitochondrial ROS production, facilitating TNBC cell proliferation and migration, effects which were mitigated by the LEPR inhibitor Allo-aca. Co-expression of LEPR and ADRB2 was observed on cell membranes, and bioinformatics data revealed a positive correlation between the two receptors. Leptin activated both LEPR and ADRB2, enhancing intracellular ROS generation and promoting tumor progression, which was effectively countered by a specific ADRB2 inhibitor ICI118551. In vivo, leptin injection accelerated tumor growth and lung metastases without affecting appetite, while treatments with Allo-aca or ICI118551 mitigated these effects. Conclusion This study demonstrates that leptin stimulates the growth and metastasis of TNBC through the activation of both LEPR and ADRB2, resulting in increased ROS production. These findings highlight LEPR and ADRB2 as potential biomarkers and therapeutic targets in TNBC.

[摘 要] 目的：探讨丹皮酚（Pae）通过调控果蝇zeste基因增强子同源物2/神经外营养蛋白4抗体/细胞周期蛋白依赖性激酶抑制因子2A（EZH2/NXPH4/CDKN2A）轴对肺癌A549细胞增殖、迁移和侵袭能力的影响。方法：以梯度浓度（0、6.25、12.5、25、50、100、200 µg/mL）的Pae处理肺癌A549细胞，采用CCK-8法确定干预浓度；将A549细胞分为对照组（未经任何处理的A549细胞）、Pae组（12.5 µg/mL Pae）、Pae + siEZH2组（转染siEZH2 + 12.5 µg/mL Pae）、Pae+siNC组（转染siNC + 12.5 µg/mL Pae）、Pae + vector NC组（转染vectorNC + 12.5 µg/mL Pae）、Pae + vectorEZH2组（转染vector EZH2+12.5 µg/mL Pae），予以相应的处理后，采用克隆形成实验检测细胞克隆数，流式细胞术检测细胞周期分布，Transwell实验检测细胞侵袭能力的变化，划痕实验检测细胞迁移能力的变化，流式细胞仪检测细胞凋亡情况，WB法检测EZH2、NXPH4、CDKN2A、Bcl-2和caspase-3蛋白表达量。在A549细胞中单独转染siEZH2或siNC，采用流式细胞仪测量细胞凋亡，WB法检测Bcl-2和caspase-3蛋白表达。建立A549细胞裸鼠移植瘤模型，评估Pae的体内抗肿瘤作用及其对相关蛋白表达的影响。结果：选择12.5 µg/mL为后续实验干预浓度。与对照组相比，Pae组细胞克隆数、S期细胞比例、迁移、侵袭能力以及EZH2、NXPH4和Bcl-2蛋白表达量显著降低，G0/G1期细胞比例、细胞凋亡率以及CDKN2A和caspase-3蛋白表达量明显升高（均P < 0.05）；与Pae + siNC组相比，Pae + siEZH2组细胞克隆数、S期细胞比例、迁移侵袭能力以及EZH2、NXPH4和Bcl-2蛋白表达量下降幅度更大，G0/G1期细胞比例、细胞凋亡率以及CDKN2A和caspase-3蛋白表达量升高幅度更大（P < 0.05）；与Pae+vectorNC组相比，Pae + vectorEZH2组细胞克隆数、S期细胞比例、迁移侵袭能力以及EZH2、NXPH4和Bcl-2蛋白表达量显著升高，G0/G1期细胞比例、细胞凋亡率以及CDKN2A和caspase-3蛋白表达量明显下降（P < 0.05）。敲低EZH2后A549细胞凋亡率和caspase-3表达明显上升，Bcl-2表达明显下降（P < 0.05）。体内实验表明Pae可显著降低肿瘤体积和质量且抑制EZH2/NXPH4/CDKN2A通路的活性。结论：Pae通过抑制EZH2/NXPH4/CDKN2A通路抑制A549细胞增殖、迁移和侵袭，诱导细胞凋亡。

[This corrects the article DOI: 10.1016/j.apsb.2022.05.019.].

Recent studies have indicated that the expression of ubiquitin-specific protease 51 (USP51), a novel deubiquitinating enzyme (DUB) that mediates protein degradation as part of the ubiquitin‒proteasome system (UPS), is associated with tumor progression and therapeutic resistance in multiple malignancies. However, the underlying mechanisms and signaling networks involved in USP51-mediated regulation of malignant phenotypes remain largely unknown. The present study provides evidence of USP51's functions as the prominent DUB in chemoresistant triple-negative breast cancer (TNBC) cells. At the molecular level, ectopic expression of USP51 stabilized the 78 kDa Glucose-Regulated Protein (GRP78) protein through deubiquitination, thereby increasing its expression and localization on the cell surface. Furthermore, the upregulation of cell surface GRP78 increased the activity of ATP binding cassette subfamily B member 1 (ABCB1), the main efflux pump of doxorubicin (DOX), ultimately decreasing its accumulation in TNBC cells and promoting the development of drug resistance both in vitro and in vivo. Clinically, we found significant correlations among USP51, GRP78, and ABCB1 expression in TNBC patients with chemoresistance. Elevated USP51, GRP78, and ABCB1 levels were also strongly associated with a poor patient prognosis. Importantly, we revealed an alternative intervention for specific pharmacological targeting of USP51 for TNBC cell chemosensitization. In conclusion, these findings collectively indicate that the USP51/GRP78/ABCB1 network is a key contributor to the malignant progression and chemotherapeutic resistance of TNBC cells, underscoring the pivotal role of USP51 as a novel therapeutic target for cancer management.

Hypercholesterolemia is a significant risk factor for the development of atherosclerosis. 2',3',5'-Tri-O-acetyl-N ⁶-(3-hydroxyphenyl) adenosine (IMM-H007), a novel AMPK agonist, has shown protective effects in metabolic diseases. However, its impact on cholesterol and triglyceride metabolism in hypercholesterolemia remains unclear. In this study, we aimed to elucidate the effects and specific mechanisms by which IMM-H007 regulates cholesterol and triglyceride metabolism. To achieve this goal, we used Apoe ^-/- and Ldlr ^-/- mice to establish a hypercholesterolemia/atherosclerosis model. Additionally, hepatocyte-specific Ampka1/2 knockout mice were subjected to a 5-week high-cholesterol diet to establish hypercholesterolemia, while atherosclerosis was induced via AAV-PCSK9 injection combined with a 16-week high-cholesterol diet. Our results demonstrated that IMM-H007 improved cholesterol and triglyceride metabolism in mice with hypercholesterolemia. Mechanistically, IMM-H007 modulated the AMPKα1/2-LDLR signaling pathway, increasing cholesterol uptake in the liver. Furthermore, IMM-H007 activated the AMPKα1-FXR pathway, promoting the conversion of hepatic cholesterol to bile acids. Additionally, IMM-H007 prevented hepatic steatosis by activating the AMPKα1/2-ATGL pathway. In conclusion, our study suggests that IMM-H007 is a promising therapeutic agent for improving hypercholesterolemia and atherosclerosis through the activation of AMPKα.

SARS-CoV-2 and its emerging variants continue to pose a significant global public health threat. The SARS-CoV-2 main protease (M^pro) is a critical target for the development of antiviral agents that can inhibit viral replication and transcription. In this study, we identified chebulagic acid (CHLA), isolated from Terminalia chebula Retz., as a potent non-peptidomimetic and non-covalent M^pro inhibitor. CHLA exhibited intermolecular interactions and provided significant protection to Vero E6 cells against a range of SARS-CoV-2 variants, including the wild-type, Delta, Omicron BA.1.1, BA.2.3, BA.4, and BA.5, with EC₅₀ values below 2 μmol/L. Moreover, in vivo studies confirmed the antiviral efficacy of CHLA in K18-hACE2 mice. Notably, CHLA bound to a unique groove at the interface between M^pro domains I and II, which was revealed by the high-resolution crystal structure (1.4 Å) of the M^pro-CHLA complex, shrinking the substrate binding pocket of M^pro and inducing M^pro aggregation. CHLA was proposed to act as an allosteric inhibitor. Pharmacokinetic profiling and safety assessments underscore CHLA's potential as a promising broad-spectrum antiviral candidate. These findings report a novel binding site on M^pro and identify antiviral activity of CHLA, providing a robust framework for lead compounds discovery and elucidating the underlying molecular mechanisms of inhibition.