OBJECTIVE: To investigate the anti-tumor effects of cinobufacini (CINO) on hepatocellular carcinoma (HCC) induced by des-gamma-carboxy-prothrombin (DCP) and to uncover the underlying mechanisms. METHODS: The inhibitory effect of CINO on HCC cell proliferation was evaluated using the cell counting kit-8 method, and the apoptosis rate was quantified using flow cytometry. Immunofluorescence and Western blot analyses were used to investigate the differential expression of proteins associated with cell growth, apoptosis, migration, and invasion pathways after CINO treatment. The therapeutic potential of CINO for HCC was confirmed, and the possibility of combining cinobufacini with c-Met inhibitor for the treatment of primary HCC was further validated by in vivo experiments. RESULTS: Under the induction of DCP, CINO inhibited the activity of HCC cells, induced apoptosis, and inhibited migration and invasion. Upon the induction of DCP, CINO regulated c-Met activation and the activation of the phosphatidylinositol-3 kinase/protein kinase B (PI3K/AKT) and mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK) pathways. In a mouse model of HCC, CINO exhibited significant antitumor effects by inhibiting the phosphorylation of c-Met and the downstream PI3K/AKT and MEK/ERK pathways in tumor tissues. CONCLUSIONS CINO inhibited HCC cell growth, promoted apoptosis, and suppressed HCC cell invasion and migration by targeting c-Met and PI3K/AKT and MEK/ERK signaling pathways under DCP induction.
Carcinoma, Hepatocellular/drug therapy* ; Proto-Oncogene Proteins c-met/metabolism* ; Liver Neoplasms/drug therapy* ; Signal Transduction/drug effects* ; Animals ; Humans ; Cell Movement/drug effects* ; Apoptosis/drug effects* ; Cell Proliferation/drug effects* ; Amphibian Venoms/therapeutic use* ; Cell Line, Tumor ; Neoplasm Metastasis ; Cell Survival/drug effects* ; Proto-Oncogene Proteins c-akt/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Neoplasm Invasiveness ; Mice, Inbred BALB C ; Mice, Nude ; Mice ; Male ; Bufanolides/therapeutic use* ; Protein Precursors ; Prothrombin ; Biomarkers

Antibiotic adjuvants offer a promising strategy for restoring antibiotic sensitivity, expanding antibacterial spectra, and reducing required dosages. Previously, compound 15 was identified as a potential adjuvant for Polymyxin B (PB) against multidrug-resistant (MDR) Pseudomonas aeruginosa DK2; however, its clinical utility was hindered by high cytotoxicity, uncertain in vivo efficacy, and an unclear synergetic mechanism. To address these challenges, we synthesized and evaluated a series of novel benzamide derivatives, with A22 emerging as a particularly promising candidate. A22 demonstrated potent synergistic activity to PB, minimal cytotoxicity, improved water solubility, and broad-spectrum synergism of polymyxins against various clinically isolated MDR Gram-negative strains. In vivo studies using Caenorhabditis elegans and mouse models further confirmed the efficacy of A22. Moreover, A22 effectively suppressed the development of PB resistance in Pseudomonas aeruginosa DK2. Mechanistic investigations revealed that A22 enhances polymyxins activity by inducing reactive oxygen species production, reducing ATP levels, increasing NOX activity, and inhibiting biofilm formation, leading to bacterial death. These findings position A22 as a highly promising candidate for the development of polymyxin adjuvants, offering a robust approach to combating MDR Gram-negative bacterial infections.

Human carboxylesterase 2A (hCES2A) plays pivotal roles in prodrug activation and hydrolytic metabolism of ester-bearing chemicals. Targeted inhibition of intestinal hCES2A represents a feasible strategy to mitigate irinotecan-triggered gut toxicity (ITGT), but the orally active, selective, and efficacious hCES2A inhibitors are rarely reported. Here, a novel drug-like hCES2A inhibitor was developed via three rounds of structure-based drug design (SBDD) and structural optimization. Initially, donepezil was identified as a moderate hCES2A inhibitor from 2000 US Food and Drug Administration (FDA)-approved drugs. Following two rounds of SBDD and structural optimization, a donepezil derivative (B7) was identified as a strong reversible hCES2A inhibitor. Subsequently, nine B7 carbamates were rationally designed, synthesized and biologically assayed. Among all synthesized carbamates, C3 showed the most potent time-dependent inhibition on hCES2A (IC₅₀ = 0.56 nmol/L), excellent specificity and favorable drug-like properties. C3 could covalently modify the catalytic serine of hCES2A with high selectivity, while this agent also showed favorable safety profiles, high intestinal exposure, and impressive effects for ameliorating ITGT in both human intestinal organoids and tumor-bearing mice. Collectively, this study showcases a rational strategy for developing drug-like and serine-targeting covalent inhibitors against target serine hydrolase(s), while C3 emerges as a promising orally active drug candidate for ameliorating ITGT.

Heatstroke, a life-threatening illness, poses a significant risk to human health, particularly in high-temperature and high-humidity environments. Timely and effective on-site management is critical for improving patient survival and prognosis. Rapid recognition, rapid assessment, and rapid cooling are the cornerstones of pre-hospital care. However, the absence of a standardized protocol for pre-hospital management of heatstroke has impeded the efficacy of treatment. This consensus, initiated by the Expert Group on Heatstroke Prevention of the People's Liberation Army, signifies a collaborative endeavor involving emergency medical personnel, nurses, and administrators from pre-hospital care, emergency departments, and intensive care units in both military and civilian domains. By systematically reviewing evidence-based medicine and clinical expertise in heatstroke prevention, on-site and in-transit care, as well as early treatment in emergency settings, the group has formulated the Expert consensus on pre-hospital emergency management of heatstroke (2024) after extensive discussions and iterative recommendations, which serve as a scientific and standardized framework for pre-hospital heatstroke emergency care. The consensus underscores the pivotal role of enhancing public awareness regarding heatstroke prevention and augmenting the rates of rapid recognition and rapid cooling for effective on-site heatstroke management. In high-risk industries, regions, or seasons for heatstroke, developing scientifically sound plans and conducting practical training can provide effective safety measures. Emergency personnel should undergo specialized training and assessments in knowledge and skills, ambulances should be equipped with effective cooling devices, and hospitals must maintain comprehensive emergency response capabilities. It is recommended to establish a regional heatstroke treatment network to optimize the allocation of emergency resources and streamline processes, thereby improving treatment outcomes and response times.
Heat Stroke/prevention & control* ; Humans ; Emergency Medical Services ; Consensus

[摘 要] 目的：制备并表征负载吲哚菁绿（ICG）的纳米胶束（F127-ICG），利用其光热效应、抗原吸附能力及淋巴结（LN）靶向优势，探索F127-ICG的抗肿瘤作用。方法：采用薄膜水化法制备F127-ICG，通过粒度电位仪测量F127-ICG的粒径及Zeta电位，通过紫外可见分光光度计及荧光分光光度计检测F127-ICG的吸收光谱及荧光光谱。通过比较F127胶束与肿瘤细胞裂解液孵育前后基本性质及蛋白含量变化，分析F127-ICG的抗原吸附作用。Calcein-AM/PI双染法检测F127-ICG对乳腺癌细胞4T1的光热杀伤作用。在小鼠皮下注射染料标记的F127胶束构建淋巴引流模型，使用小动物活体成像检测F127胶束的LN靶向作用，并使用离体器官成像检测F127胶束在小鼠腹股沟LN及腋窝LN中蓄积和渗透情况。构建小鼠背部双侧乳腺癌肿瘤模型，小鼠瘤内注射F127-ICG进行光热治疗，观察对侧肿瘤生长趋势。结果：成功构建负载ICG的F127胶束，粒径为（19.41 ± 0.49）nm，Zeta电位为-（2.78 ± 0.36）mV。F127-ICG与肿瘤抗原共孵育后粒径、负Zeta电位以及蛋白含量均增大（P < 0.05）。Calcein-AM/PI双染法结果显示，F127-ICG可以发挥光热效应杀伤4T1细胞。活体成像结果显示，F127胶束可靶向LN。动物体内实验中，与PBS及F127-ICG组相比，F127-ICG + 激光组的对侧肿瘤体积更小（P < 0.05）。结论：F127-ICG通过光热消融原位肿瘤组织，同时捕获释放的肿瘤抗原并迁移至局部LN，促进机体抗肿瘤免疫应答，抑制远处肿瘤生长，增强原位疫苗效应。