The underlying cause of low response rates to existing immunotherapies is that tumor cells dominate tumor immune escape through surface antigen deficiency and inducing tumor immunosuppressive microenvironment (TIME). Here, we proposed an in situ tumor cell engineering strategy to disrupt tumor immune escape at the root by restoring tumor cell MHC-I/tumor-specific antigen complex (MHC-I/TSA) expression to promote T-cell recognition and by silencing tumor cell CD55 to increase the ICOSL⁺ B-cell proportion and reverse the TIME. A doxorubicin (DOX) and dual-gene plasmid (MAC pDNA, encoding both MHC-I/ASMTNMELM and CD55-shRNA) coloaded drug delivery system (LCPN@ACD) with tumor targeting and charge/size dual-conversion properties was prepared. LCPN@ACD-induced ICD promoted DC maturation and enhanced T-cell activation and infiltration. LCPN@ACD enabled effective expression of MHC-I/TSA on tumor cells, increasing the ability of tumor cell recognition and killing. LCPN@ACD downregulated tumor cell CD55 expression, increased the proportion of ICOSL⁺ B cells and CTLs, and reversed the TIME, thus greatly improving the efficacy of αPD-1 and CAR-T therapies. The application of this in situ tumor cell engineering strategy eliminated the source of tumor immune escape, providing new ideas for solving the challenges of clinical immunotherapy.

Due to their good properties of elastic modulus, degradability and ability to promote bone repair, magnesium alloys have become a research hotspot in research of orthopedic implants. Nevertheless, most of the biomedical magnesium alloys currently available fail to meet the requirements in orthopedics because of their rapid degradation after implantation. Rare earth-magnesium alloys possess excellent corrosion resistance and are expected to become important materials as clinical orthopedic implants. This review summarizes the recent progress in studies of the physiological functions of rare earth elements, the effects of supplementation of rare earth elements on biomechanical properties and the in vitro and in vivo biocompatibility of magnesium alloys, and their contribution to tendon-bone healing, addressing also the current clinical orthopedic applications of different rare earth-magnesium alloys, challenges, and future strategies for improving these alloys.
Alloys/chemistry* ; Magnesium/chemistry* ; Metals, Rare Earth/chemistry* ; Humans ; Biocompatible Materials ; Prostheses and Implants

Objective·To optimize iSeroSnFR,a genetically encoded 5-hydroxytryptamine(5-HT)fluorescent sensor based on bacterial periplasmic binding proteins(PBPs),to enhance its performance for both in vivo and in vitro 5-HT detection.Methods·iSeroSnFR1.2 was engineered by replacing the circularly permuted superfolder green fluorescence protein(cpsfGFP)sequence in iSeroSnFR1.0 with that from the acetylcholine sensor iAChSnFR using Gibson assembly.The fluorescence response and kinetic properties of iSeroSnFR1.0 and iSeroSnFR1.2 were compared by overexpressing the sensors in HEK293 cells and puffing with exogenous 5-HT.Additionally,to mimic physiological conditions,cultured mouse cortical neurons infected with Sindbis virus carrying each sensor were electrically stimulated to induce endogenous 5-HT release and further evaluate sensor performance.Results·iSeroSnFR1.2 showed significantly improved performance over iSeroSnFR1.0.In HEK293 cells,it exhibited a 1.5-fold increase in fluorescence response(ΔF/F0)to exogenous 5-HT,along with faster kinetics(rise time:36.3 ms vs 44.9 ms;decay time:1 003.6 ms vs 1 730.4 ms).In cortical neurons,it demonstrated a 2.7-fold increase in response to endogenously released 5-HT,with rise and decay times reduced by 44.0%and 26.7%,respectively.Notably,iSeroSnFR1.2 showed increased basal fluorescence,enabling better imaging in high-background environments.Conclusion·The optimized iSeroSnFR1.2 sensor offers a markedly improved fluorescent response and temporal resolution for 5-HT detection,providing an advanced tool for studying 5-HT dynamics in neuroscience and psychiatric research.

Stresses induced by the deficiency or excess of trace mineral elements, such as manganese (Mn), represent a common limiting factor for the production of crops like Brassica napus. To identify key genes involved in Mn allocation in B. napus and elucidate the underlying mechanisms, a member of the metal tolerance protein (MTP) family obtained in the previous screening of cDNA library of B. napus under Mn stress was selected as the research subject. Based on the sequence information and phylogenetic analysis, it was named as BnMTP10. It belongs to the Mn-cation diffusion facilitator (CDF) subfamily. Expression of BnMTP10 in yeast significantly improved the tolerance of transformants to excessive Mn and iron (Fe) and reduced the accumulation of Mn and Fe. However, the yeast transformants exhibited no significant changes in tolerance to excess cadmium, boron, aluminum, zinc, or copper. The qRT-PCR results demonstrated that the flowers of B. napus had the highest expression of BnMTP10, followed by roots and leaves. Subcellular localization studies revealed that BnMTP10 was localized in the endoplasmic reticulum (ER). Compared with wild-type plants, transgenic Arabidopsis overexpressing BnMTP10 exhibited enhanced tolerance to excessive Mn stress but showed no significant difference under Fe stress. Correspondingly, under excessive Mn stress, the Mn content in the roots of transgenic Arabidopsis increased significantly. However, under excessive Fe stress, the Fe content in transgenic Arabidopsis did not alter significantly. According to the results, we hypothesize that BnMTP10 may alleviate excessive Mn stress in plants by mediating Mn transport to the ER. This study facilitated our understanding of efficient mineral nutrients, and provided theoretical foundations and gene resources for breeding B. napus.
Brassica napus/genetics* ; Manganese/metabolism* ; Plants, Genetically Modified/genetics* ; Plant Proteins/physiology* ; Arabidopsis/metabolism* ; Gene Expression Regulation, Plant ; Phylogeny ; Cation Transport Proteins/metabolism* ; Stress, Physiological

Objective·To optimize iSeroSnFR,a genetically encoded 5-hydroxytryptamine(5-HT)fluorescent sensor based on bacterial periplasmic binding proteins(PBPs),to enhance its performance for both in vivo and in vitro 5-HT detection.Methods·iSeroSnFR1.2 was engineered by replacing the circularly permuted superfolder green fluorescence protein(cpsfGFP)sequence in iSeroSnFR1.0 with that from the acetylcholine sensor iAChSnFR using Gibson assembly.The fluorescence response and kinetic properties of iSeroSnFR1.0 and iSeroSnFR1.2 were compared by overexpressing the sensors in HEK293 cells and puffing with exogenous 5-HT.Additionally,to mimic physiological conditions,cultured mouse cortical neurons infected with Sindbis virus carrying each sensor were electrically stimulated to induce endogenous 5-HT release and further evaluate sensor performance.Results·iSeroSnFR1.2 showed significantly improved performance over iSeroSnFR1.0.In HEK293 cells,it exhibited a 1.5-fold increase in fluorescence response(ΔF/F0)to exogenous 5-HT,along with faster kinetics(rise time:36.3 ms vs 44.9 ms;decay time:1 003.6 ms vs 1 730.4 ms).In cortical neurons,it demonstrated a 2.7-fold increase in response to endogenously released 5-HT,with rise and decay times reduced by 44.0%and 26.7%,respectively.Notably,iSeroSnFR1.2 showed increased basal fluorescence,enabling better imaging in high-background environments.Conclusion·The optimized iSeroSnFR1.2 sensor offers a markedly improved fluorescent response and temporal resolution for 5-HT detection,providing an advanced tool for studying 5-HT dynamics in neuroscience and psychiatric research.

Objective:To summarize the clinical, pathological and molecular characteristics of various types of pediatric glioma, and to explore the differences in the morphology and clinical significance among various types of pediatric glioma.Methods:Based on the fifth edition of the World Health Organization classification of central nervous system tumors, this study classified or reclassified 111 pediatric gliomas that were diagnosed at Guangzhou Medical University Affiliated Women and Children′s Medical Center from January 2020 to June 2023. The clinical manifestations, imaging findings, histopathology, and molecular characteristics of these tumors were analyzed. Relevant literature was also reviewed.Results:The 111 patients with pediatric glioma included 56 males and 55 females, with the age ranging from 10 days to 13 years (average age, 5.5 years). Clinically, manifestations presented from 5 days to 8 years before the diagnosis, including epilepsy in 16 cases, increased intracranial pressure in 48 cases and neurological impairment in 66 cases. MRI examinations revealed tumor locations as supratentorial in 43 cases, infratentorial in 65 cases, and spinal cord in 3 cases. There were 73 cases presented with a solid mass and 38 cases with cystic-solid lesions. The largest tumor diameter ranged from 1.4 to 10.6 cm. Among the 111 pediatric gliomas, there were 6 cases of pediatric diffuse low-grade glioma (pDLGG), 63 cases of circumscribed astrocytoma glioma (CAG), and 42 cases of pediatric diffuse high-grade glioma (pDHGG). Patients with pDLGG and CAG were younger than those with pDHGG. The incidence of pDLGG and CAG was significantly lower in the midline of the infratentorial region compared to that of pDHGG. They were more likely to be completely resected surgically. The pDLGG and CAG group included 4 cases of pleomorphic xanthoastrocytoma, showing histological features of high-grade gliomas. Among the high-grade gliomas, 13 cases were diffuse midline gliomas and also showed histological features of low-grade glioma. Immunohistochemical studies of H3K27M, H3K27ME3, p53, ATRX, BRAF V600E, and Ki-67 showed significant differences between the pDLGG and CAG group versus the pDHGG group ( P<0.01). Molecular testing revealed that common molecular variations in the pDLGG and CAG group were KIAA1549-BRAF fusion and BRAF V600E mutation, while the pDHGG group frequently exhibited mutations in HIST1H3B and H3F3A genes, 1q amplification, and TP53 gene mutations. With integrated molecular testing, 2 pathological diagnoses were revised, and the pathological subtypes of 35.3% (12/34) of the pediatric gliomas that could not be reliably classified by histology were successfully classified. Conclusions:There are significant differences in clinical manifestations, pathological characteristics, molecular variations, and prognosis between the pDLGG, CAG and pDHGG groups. The integrated diagnosis combining histology and molecular features is of great importance for the accurate diagnosis and treatment of pediatric gliomas.

Cytopharmaceutical based on macrophages is a breakthrough in the field of targeted drug delivery. However, it remains a challenge to localize and control drug release while retaining macrophage activity and exerting its immunotherapeutic effect. Herein, a localized light-triggered release macrophage cytopharmaceutical (USIP@M) was proposed, which could utilize the tumor targeting and immunotherapy effects of macrophages to reverse the immune suppression of tumor microenvironment (TME). Amphiphilic block copolymers with ultraviolet (UV)-responsive o-nitrobenzyl groups were synthesized and co-loaded with sorafenib (SF), IMD-0354 (IMD), and upconverting nanoparticles (UCNPs), which were then taken up by macrophages, and the targeted delivery of drugs was realized by using the tumor tropism of macrophages. UCNPs converted near-infrared light with strong penetrability and high safety into UV light, which promoted the photoresponsive depolymerization of block copolymers and production of exosomes from USIP@M, accelerated drug efflux and maintained the activity of macrophages. IMD simultaneously polarized carrier macrophages and tumor-associated macrophages to exert the antitumor effect of macrophages, enhance T cell immunity, and alleviate the immunosuppressive state of TME. Synergistically with the chemotherapeutic effect of SF, it could effectively kill tumors. In conclusion, based on the localized light-triggered release strategy, this study constructed a novel macrophage cytopharmaceutical that could localize and control drug release while retaining the activity of macrophages and exerting its immunotherapeutic effect, which could effectively treat solid tumors.

ObjectiveTo identify and clarify the key issues faced by tertiary hospitals in responding to public health emergencies. MethodsA literature review index system was constructed， and key issues were identified using hierarchical analysis. ResultsAfter a systematic literature review， 20 types of problems faced by tertiary hospitals in responding to public health emergencies were identified. Three key issues were ultimately identified by prioritizing the issues that needed to be addressed. ConclusionThe key issues of tertiary hospitals in responding to public health emergencies are concentrated in the areas of emergency response capabilities and competencies of medical staff， the number of emergency response personnel， and the standardization and specificity of training and drills. Tertiary hospitals should focus on these issues in developing public health emergency response systems to improve the effectiveness of their emergency response.

ObjectiveTo identify the key issues in emergency response to public health emergencies in the Yangtze River Delta region， and to provide a basis for the formulation of relevant policies. MethodLiterature on emergency response to public health emergencies in the Yangtze River Delta region was systematically reviewed and collected， and 18 main problems were identified after collecting and organizing the problems. A questionnaire survey and cluster analysis were used to identify the key issues. ResultsThe study of 18 major problems revealed that "insufficient reserve of emergency medical supplies"， "insufficient financial investment to meet the needs of emergency response"， "lack of a sound mechanism for the deployment of supplies， resulting in the untimely deployment of materials"， were the key issues of emergency response to public health emergencies in the Yangtze River Delta region. ConclusionFuture development of the emergency response system in the Yangtze River Delta region should focus on accelerating the integration process of the Yangtze River Delta， establishing and improving information systems， optimizing the material storage and deployment network， prioritizing the fund investment and utilization mechanism， and improving the effectiveness of public health emergency response.

AIM: To reveal that the targeted regulation of TGF-β1 by miR-21-5P is the key mechanism that mediates the activation of myocardial fibroblasts, and to clarify the intervention ofRadix Angelica Sinensis and Radix Hedysari ultrafiltration on the mechanism of miR-21-5P targeting to regulate TGF-β1 effect. METHODS: (1) The cells were randomly divided into normal group and irradiation group. The irradiation group received 6Gry single irradiation, and then RT-PCR was used to detect miR-21-5P, and Western Blot was used to detect the expression of α-SMA and TGF-β1. (2) The cells were randomly divided into normal group, irradiation group, miR-21-5P