Multidrug-resistant (MDR) bacterial infections have emerged as a serious challenge of global public health crisis. The overuse and misuse of conventional antibiotics have dramatically accelerated the emergence, evolution and worldwide spread of drug-resistant bacterial strains, necessitating urgent exploration of novel antibacterial strategies. Bacteriophages serve as natural bacterial predators offering distinct advantages including high host specificity, autonomous self-replication capabilities and cost-effective large-scale production. However, wild-type phages present significant clinical limitations due to their narrow host ranges, susceptibility to rapid immune clearance and poor penetration of bacterial biofilms, which severely restrict their therapeutic applications. The convergence of synthetic biology, nanotechnology and advanced gene editing technologies has accelerated the development of engineered bacteriophage platforms, providing programmable, scalable and clinically translatable pathways to overcome these inherent biological constraints. Here, we systematically delineate four fundamental strategies for engineered bacteriophage development. Chemical modification utilizes reactive functional groups such as amino, carboxyl and thiol moieties on capsid proteins through esterification, amidation or click chemistry reactions to achieve precise drug conjugation and surface functionalization. In vivo editing encompasses ultraviolet or chemical mutagenesis for random mutation induction, homologous recombination for targeted genetic alterations, recombineering methodologies including electroporation-mediated bacteriophage recombination engineering, and CRISPR-Cas systems for precise genome editing to enable exact genetic reconstruction and host range reprogramming. In vitro synthesis leverages genome engineering platforms where intact phage genomes are transferred into yeast or host bacteria to facilitate highly efficient homologous recombination, enabling large DNA fragment assembly and cross-gene host range expansion without bacterial toxicity constraints. Directed evolution combines artificial selection through mutation library screening with rational design approaches involving chimeric receptor binding protein construction or site-specific mutagenesis, effectively balancing the discovery of unknown adaptive pathways with targeted host specificity modification. Moreover, we comprehensively discuss therapeutic applications across diverse clinical scenarios. Engineered bacteriophage effectively disrupt bacterial biofilms through sophisticated functionalized delivery platforms including nanozyme-conjugated phages, phage-liposome nanoconjugates and bio-responsive hydrogels, demonstrating significantly enhanced bactericidal efficiency compared to unmodified free phages. These bioengineered vectors attenuate bacterial virulence and resensitize pathogens to antibiotics by delivering CRISPR-Cas systems or base editors to disrupt critical virulence factors such as pili, capsule synthesis machineries and quorum sensing systems, or by inactivating antibiotic resistance determinants including beta-lactamase genes. As an intelligent nanomedicine delivery platform, engineered bacteriophage enable precise pathogen elimination an through photocatalytic reactive oxygen species generation, immunomodulatory interventions, or controlled release of antibacterial drugs. Furthermore, oral administration of engineered bacteriophage facilitates microbiota modulation, which selectively eliminate intestinal pathogens while preserve beneficial commensal microbiota, thereby restoring microbial community balance and preventing complications associated with dysbiosis. Finally, we critically analyze persistent challenges including host strain matching complexity, evolution of bacterial resistance mechanisms, pharmacokinetic optimization requirements, optimal administration route selection, large-scale production quality control standards and clinical dosing determination protocols. Through multidisciplinary integration of synthetic biology, infectious disease medicine and immunology, future translational medicine studies of bacteriophage should establish comprehensive technical platforms encompassing rapid phage screening, intelligent rational design, rigorous in vivo evaluation and standardized clinical validation processes, ultimately advancing engineered bacteriophage from laboratory innovations to clinically approved therapeutics for effectively combating MDR bacterial infections.

Nuclear receptor co-activator 4 (NCOA4) acts as a selective cargo receptor that binds to ferritin, a cytoplasmic iron storage complex. By mediating ferritinophagy, NCOA4 regulates iron metabolism and releases free iron in the body, thus playing a crucial role in a variety of biological processes, including growth, development, and metabolism. Recent studies have shown that NCOA4-mediated ferritinophagy is closely associated with the occurrence and development of iron metabolism-related diseases, such as liver fibrosis, renal cell carcinoma, and neurodegenerative diseases. In addition, a number of clinical drugs have been identified to modulate NCOA4-mediated ferritinophagy, significantly affecting disease progression and treatment efficacy. This paper aims to review the current research progress on the role of NCOA4-mediated ferritinophagy in related diseases, in order to provide new ideas for targeted clinical therapy.
Humans ; Nuclear Receptor Coactivators/physiology* ; Ferritins/metabolism* ; Animals ; Neurodegenerative Diseases/metabolism* ; Iron/metabolism* ; Autophagy/physiology* ; Liver Cirrhosis/metabolism* ; Carcinoma, Renal Cell/metabolism* ; Kidney Neoplasms/physiopathology*

The dried mature peel of Citrus reticulata, a plant in the Rutaceae family and its cultivated varieties, is a commonly used Chinese medicinal material known as Chenpi(Citri Reticulatae Pericarpium). It is rich in nutritional components and medicinal value, with pharmacological effects including relieving cough and eliminating phlegm, strengthening the spleen and drying dampness, protecting the liver and benefiting the stomach, tonifying Qi, and calming the mind. Huanglongbing(HLB), also known as Citrus Huanglongbing, is a destructive disease in citrus production that seriously threatens the development of the citrus industry. HLB causes symptoms such as the inability of Rutaceae plants to produce mature fruit, gradual weakening of the tree, and eventual death, posing a significant threat to the yield and quality of Chenpi. Due to the uneven distribution of the HLB pathogen in infected plants, accurate detection of the pathogen requires the collection of a large number of plant samples. Current sample pretreatment methods, such as traditional extraction methods and commercial extraction kits, are time-consuming and involve multiple steps, which significantly increase the difficulty and workload of HLB diagnosis and have become a bottleneck in HLB detection. In this study, a rapid high-throughput detection method combining alkali lysis and TaqMan qPCR was developed. This method allows the pretreatment of multiple samples within 5 min, and the entire detection process can be completed within 45 min, with a detection limit of 6.67 fg·μL~(-1). The alkali lysis method and commercial kits were used for parallel detection of field-collected citrus samples, and the results showed no significant difference. The sample pretreatment method established in this study is characterized by low cost, simplicity, and high efficiency. Combined with TaqMan qPCR, it can provide technical support for early and on-site diagnosis of HLB. This method is of great significance for disease prevention and control in the citrus industry and is expected to help improve the yield and quality of citrus medicinal materials.
Citrus/microbiology* ; Plant Diseases/microbiology* ; Rhizobiaceae/physiology* ; High-Throughput Screening Assays/methods* ; Liberibacter/physiology*

In recent years,significant breakthroughs have been achieved in the immunotherapy for Alzheimer's disease.In line with global advancements,two anti-amyloid-β monoclonal antibodies have been approved and successfully launched in China for clinical use.Lecanemab and Donanemab were officially used in June 2024 and April 2025 in China,respectively.In order to standardize the rational and safe application of anti-amyloid-β monoclonal antibodies for Alzheimer's disease in China,this article integrates recom-mendations from the clinical trials and real-world experience from the author's team and domestic peers to further update the recom-mendations for the clinical use of anti-amyloid-β monoclonal antibody based on the 2024 version.It includes indications for therapy,pre-treatment evaluation and preparation,administration protocols and safety measures during treatment,and post-treatment monitor-ing strategies.

OBJECTIVE: To investigate the effect of miR-483-5p on hepatocellular carcinoma (HCC) cells proliferation and stemness, as well as the attenuating effect of Pien Tze Huang (PZH). METHODS: Differentially expressed miRNA between HepG2 cells and hepatic cancer stem-like cells (HCSCs) were identified by a miRNA microarray assay. miR-483-5p mimics were transfected into HepG2 cells to explore the effects of miR-483-5p on cell proliferation and stemness. HepG2 cells and HCSCs were treated with PZH (0, 0.25, 0.50 and 0.75 mg/mL) to explore the effects of PZH on the proliferation and stemness, both in non-induced state and the state induced by miR-483-5p mimics. RESULTS: miR-483-5p was significantly up-regulated in HCSCs and its overexpression increased cell proliferation and stemness in HepG2 cells (P<0.05). PZH not only significantly inhibited proliferation in HepG2 cells, but also significantly suppressed the cell proliferation and self-renewal of HCSCs (P<0.05). The effects of miR-483-5p mimics on proliferation and stemness of HepG2 cells were partially abolished by PZH. CONCLUSIONS miR-483-5p promotes proliferation and enhances stemness of HepG2 cells, which were attenuated by PZH, demonstrating that miR-483-5p is a potential molecular target for the treatment of HCC and provide experimental evidence to support clinical use of PZH for patients with HCC.
Humans ; MicroRNAs/metabolism* ; Cell Proliferation/drug effects* ; Liver Neoplasms/drug therapy* ; Carcinoma, Hepatocellular/drug therapy* ; Hep G2 Cells ; Neoplastic Stem Cells/metabolism* ; Drugs, Chinese Herbal/therapeutic use* ; Gene Expression Regulation, Neoplastic/drug effects*

AIM To investigate the effects of Heixiaoyao Powder on neuroinflammation in APP/PS1 transgenic mice.METHODS The 16-week-old male APP/PS1 transgenic mice were randomly divided into the model group,the BBG group(P2X7R specific antagonist,30 mg/kg)and high,medium and low dose Heixiaoyao Powder groups of(22.10,11.05,5.53 g/kg),in contrast to the male C57BL/6J mice of the same age and the same strain of the blank groups,with 12 mice in each group.When normal saline by gavage was dosed upon the blank group and the model group,and the other groups were treated with corresponding drug by gavage.After 90 days of administration,the mice had their learning and memory ability detected by Morris water maze test;their hippocampal pathological changes observed with HE staining;their MyD88 expression detected by immunofluorescence staining;their hippocampal levels of pro-inflammatory factors(TNF-α,IL-6),anti-inflammatory factors(IL-10),ATP and amyloid protein β(Aβ)detected by ELISA;their hippocampal mRNA expressions of P2X7R,TLR4,MyD88 and NF-κB-P65 detected by RT-qPCR method;and their hippocampal protein expressions of P2X7R,TLR4,MyD88,NF-κB-P65 and p-NF-κB-P65 detected by Western blot.RESULTS Compared with the blank group,the model group demonstrated prolonged escape latency and reduced frequency in crossing platform(P＜0.01);decreased number of hippocampal neurons,deranged neurons,and darker cytoplasm staining;increased immunofluorescence expression of hippocampal MyD88(P＜0.01);decreased IL-10 level(P＜0.01);increased levels of TNF-α,IL-6,ATP and Aβ(P＜0.01);increased mRNA and protein expressions ofP2X7R,TLR4 and MyD88(P＜0.01),and increased protein expression of p-NF-κB-P65(P＜0.01).Compared with the model group,the groups intervened with Heixiaoyao Powder or BBG demonstrated shortened escape latency and increased frequency in crossing platform(P＜0.01);more number of neatly arranged hippocampal neurons;increased hippocampal IL-10 level(P＜0.01),decreased levels of TNF-α,IL-6,ATP and Aβ(P＜0.05,P＜0.01);decreased mRNA and protein expressions of P2X7R,TLR4 and MyD88(P＜0.05,P＜0.01);and decreased protein expression of p-NF-κB-P65(P＜0.05,P＜0.01).Except the low dose Heixiaoyao Powder group,the other treatment groups shared decreased immunofluorescence expression of MyD88(P＜0.05,P＜0.01).CONCLUSION Heixiaoyao Powder can significantly improve the learning and memory ability of APP/PS1 model mice,and its mechanism may lie in its function in alleviating cerebral neuroinflammation by reducing the abnormal Aβ aggregation via inhibiting the activation of ATP/P2X7R/NF-κB signaling pathway.

A mounting body of research suggests that circRNAs significantly contribute to the develop-ment of myocardial fibrosis.The microarray results of human circular RNA expression profile indicated that circHERC4_041 expression increased in the myocardium of patients with heart failure,RT-qPCR a-nalysis confirmed that the myocardial expression level of circHERC4_041 in individuals with heart failure were considerably elevated compared to that in healthy organ donors.Fluorescence in situ hybridization(FISH)confirmed that circHERC4_041 was abundant in the cytoplasm of human cardiomyocyte AC16.Overexpression of circHERC4_041 in mouse myocardial fibroblasts(mCFs)mediated by adenovirus in-hibited the expression of fibrosis-related proteins in mCFs.Experiments involving cell proliferation,wound healing,and Transwell assays demonstrated that overexpression of circHERC4_041 suppressed the growth and mobility of mCFs(P＜0.001).Sequence analysis results suggested that circHERC4_041 con-tains potential ribosome entry sequence(IRES)and open reading frame(ORF).Western blot confirmed that circHERC4_041 could translate the 516 amino acid HERC4-516aa protein,which was mainly located in the cytoplasm of the cell.Cell functional experiments confirmed that circHERC4_041 inhibited the fi-brotic phenotype of mCFs by specifically translating HERC4-516aa(P＜0.05).The specific interaction between HERC4-516aa and transglutaminase 2(TGM2)was confirmed by IP-MS screening and Co-IP i-dentification.Further results found that the degradation of TGM2 was promoted through proteasome path-way.The overexpression of TGM2 in mCFs facilitated by adenoviral vectors could counteract the suppres-sive effects of HERC4-516aa on the fibrotic phenotype of mCFs.Therefore,this study confirmed that the HERC4-516aa protein translated by circHERC4_041 can specifically bind to TGM2 to inhibit the fibrotic phenotype of myocardial fibroblasts.

A mounting body of research suggests that circRNAs significantly contribute to the develop-ment of myocardial fibrosis.The microarray results of human circular RNA expression profile indicated that circHERC4_041 expression increased in the myocardium of patients with heart failure,RT-qPCR a-nalysis confirmed that the myocardial expression level of circHERC4_041 in individuals with heart failure were considerably elevated compared to that in healthy organ donors.Fluorescence in situ hybridization(FISH)confirmed that circHERC4_041 was abundant in the cytoplasm of human cardiomyocyte AC16.Overexpression of circHERC4_041 in mouse myocardial fibroblasts(mCFs)mediated by adenovirus in-hibited the expression of fibrosis-related proteins in mCFs.Experiments involving cell proliferation,wound healing,and Transwell assays demonstrated that overexpression of circHERC4_041 suppressed the growth and mobility of mCFs(P＜0.001).Sequence analysis results suggested that circHERC4_041 con-tains potential ribosome entry sequence(IRES)and open reading frame(ORF).Western blot confirmed that circHERC4_041 could translate the 516 amino acid HERC4-516aa protein,which was mainly located in the cytoplasm of the cell.Cell functional experiments confirmed that circHERC4_041 inhibited the fi-brotic phenotype of mCFs by specifically translating HERC4-516aa(P＜0.05).The specific interaction between HERC4-516aa and transglutaminase 2(TGM2)was confirmed by IP-MS screening and Co-IP i-dentification.Further results found that the degradation of TGM2 was promoted through proteasome path-way.The overexpression of TGM2 in mCFs facilitated by adenoviral vectors could counteract the suppres-sive effects of HERC4-516aa on the fibrotic phenotype of mCFs.Therefore,this study confirmed that the HERC4-516aa protein translated by circHERC4_041 can specifically bind to TGM2 to inhibit the fibrotic phenotype of myocardial fibroblasts.

AIM To investigate the effects of Heixiaoyao Powder on neuroinflammation in APP/PS1 transgenic mice.METHODS The 16-week-old male APP/PS1 transgenic mice were randomly divided into the model group,the BBG group(P2X7R specific antagonist,30 mg/kg)and high,medium and low dose Heixiaoyao Powder groups of(22.10,11.05,5.53 g/kg),in contrast to the male C57BL/6J mice of the same age and the same strain of the blank groups,with 12 mice in each group.When normal saline by gavage was dosed upon the blank group and the model group,and the other groups were treated with corresponding drug by gavage.After 90 days of administration,the mice had their learning and memory ability detected by Morris water maze test;their hippocampal pathological changes observed with HE staining;their MyD88 expression detected by immunofluorescence staining;their hippocampal levels of pro-inflammatory factors(TNF-α,IL-6),anti-inflammatory factors(IL-10),ATP and amyloid protein β(Aβ)detected by ELISA;their hippocampal mRNA expressions of P2X7R,TLR4,MyD88 and NF-κB-P65 detected by RT-qPCR method;and their hippocampal protein expressions of P2X7R,TLR4,MyD88,NF-κB-P65 and p-NF-κB-P65 detected by Western blot.RESULTS Compared with the blank group,the model group demonstrated prolonged escape latency and reduced frequency in crossing platform(P＜0.01);decreased number of hippocampal neurons,deranged neurons,and darker cytoplasm staining;increased immunofluorescence expression of hippocampal MyD88(P＜0.01);decreased IL-10 level(P＜0.01);increased levels of TNF-α,IL-6,ATP and Aβ(P＜0.01);increased mRNA and protein expressions ofP2X7R,TLR4 and MyD88(P＜0.01),and increased protein expression of p-NF-κB-P65(P＜0.01).Compared with the model group,the groups intervened with Heixiaoyao Powder or BBG demonstrated shortened escape latency and increased frequency in crossing platform(P＜0.01);more number of neatly arranged hippocampal neurons;increased hippocampal IL-10 level(P＜0.01),decreased levels of TNF-α,IL-6,ATP and Aβ(P＜0.05,P＜0.01);decreased mRNA and protein expressions of P2X7R,TLR4 and MyD88(P＜0.05,P＜0.01);and decreased protein expression of p-NF-κB-P65(P＜0.05,P＜0.01).Except the low dose Heixiaoyao Powder group,the other treatment groups shared decreased immunofluorescence expression of MyD88(P＜0.05,P＜0.01).CONCLUSION Heixiaoyao Powder can significantly improve the learning and memory ability of APP/PS1 model mice,and its mechanism may lie in its function in alleviating cerebral neuroinflammation by reducing the abnormal Aβ aggregation via inhibiting the activation of ATP/P2X7R/NF-κB signaling pathway.