Objective To evaluate the safety and efficacy of a self-developed micro-normothermic machine perfusion (NMP) system (micro-perfusion device) for preserving isolated porcine limbs. Methods Five healthy Landrace pigs were selected, and their left and right forelimbs were randomly divided into the NMP group and static cold storage (SCS) group. The NMP group was perfused with the self-developed micro-perfusion device and polymerized hemoglobin perfusate for 32 hours at normothermia, while the SCS group was preserved at 4 ℃. Hemodynamic parameters such as perfusion pressure and flow were monitored. The pH value, partial pressure of oxygen (PO₂), lactic acid (Lac), creatine kinase (CK) and lactate dehydrogenase (LDH) in the perfusate were measured. Hematoxylin-eosin staining was used to assess the muscle tissue structure, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was employed to evaluate muscle cell apoptosis, and immunohistochemistry staining was applied to detect the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6. A mixed-effects model was used to analyze the effects of time and treatment methods on tissue structure, cell apoptosis and inflammatory factors. Results The device could stably maintain a perfusion pressure of (69±15) mmHg and a flow rate of (117±42) mL/min. The pH value and electrolytes of the perfusate were generally stable, with PO₂ maintained at a high level. Lac was maintained at 5.38(3.81, 6.45) mmol/L, while CK and LDH increased over time. After 32 hours of perfusion in the NMP group, both the myocyte spacing and apoptosis rate were better than those in the SCS group. Mixed-effects model analysis showed that there were statistically significant differences in the effects of NMP treatment and SCS treatment on myocyte spacing and apoptosis rate per unit time (both P < 0.05). There were no statistically significant differences in TNF-α and IL-6 between the two groups, and mixed-effects model analysis showed no statistically significant differences in the effects of NMP treatment and SCS treatment on TNF-α and IL-6 per unit time (both P > 0.05). Conclusions The micro-perfusion device used in this study may achieve 32-hour normothermic preservation in a porcine limb amputation model, maintain basic metabolism and ionic homeostasis, reduce muscle structural damage and cell apoptosis without inducing additional inflammatory responses. This technology is expected to significantly extend the time window for replantation of amputated limbs in disaster rescue and long-distance transportation, providing an important technical basis for clinical translation and subsequent replantation research.

Acute lung injury （ALI） is a life-threatening condition characterized by uncontrolled pulmonary inflammatory responses， presenting high morbidity and mortality rates that pose serious threats to patient health. The pathogenesis of ALI involves complex interactions among pathological mechanisms including immune-inflammatory responses， disruption of the alveolar-capillary barrier， mesenchymal stem cell dysfunction， metabolic disorders， ferroptosis， and alterations in the gut microbiota. The dysregulation of programmed cell death （PCD） has increasingly emerged as a research focus. PCD represents a genetically regulated autonomous process of cell death， crucial for maintaining the internal stability of organisms. Recent studies indicate that multiple PCD modes， including apoptosis， pyroptosis， necroptosis， autophagy， and ferroptosis， participate in the development and progression of ALI by mediating pathological processes such as alveolar epithelial cell injury and vascular endothelial barrier disruption. Current therapeutic approaches for ALI encompass lung-protective ventilation， body fluid management， mesenchymal stem cell therapy， and pharmacological interventions. While these measures may alleviate clinical symptoms， they remain unable to reverse the pathological progression of lung injury. Traditional Chinese medicine （TCM） possesses the unique advantages of a holistic perspective and syndrome differentiation-based treatment， achieving breakthrough progress in targeted PCD intervention for ALI. It can mitigate pulmonary tissue injury by suppressing inflammatory responses， regulating immune function， and improving coagulation disorders， thereby delaying the onset and progression of ALI. This paper delves into the molecular mechanisms of PCD in ALI， systematically reviewing the effects of TCM extracts， active constituents （terpenoids， saponins， flavonoids， and phenols）， and compound formulas （e.g.， Qingjie Huagong prescription， Maxing Kugan Tang， and Yifei Jianpi prescription） in improving pulmonary epithelial cell function and mitigating lung injury by modulating PCD pathways. This review aims to provide theoretical underpinnings and novel directions for drug development in the clinical prevention and treatment of ALI.

Acute lung injury （ALI） is a life-threatening condition characterized by uncontrolled pulmonary inflammatory responses， presenting high morbidity and mortality rates that pose serious threats to patient health. The pathogenesis of ALI involves complex interactions among pathological mechanisms including immune-inflammatory responses， disruption of the alveolar-capillary barrier， mesenchymal stem cell dysfunction， metabolic disorders， ferroptosis， and alterations in the gut microbiota. The dysregulation of programmed cell death （PCD） has increasingly emerged as a research focus. PCD represents a genetically regulated autonomous process of cell death， crucial for maintaining the internal stability of organisms. Recent studies indicate that multiple PCD modes， including apoptosis， pyroptosis， necroptosis， autophagy， and ferroptosis， participate in the development and progression of ALI by mediating pathological processes such as alveolar epithelial cell injury and vascular endothelial barrier disruption. Current therapeutic approaches for ALI encompass lung-protective ventilation， body fluid management， mesenchymal stem cell therapy， and pharmacological interventions. While these measures may alleviate clinical symptoms， they remain unable to reverse the pathological progression of lung injury. Traditional Chinese medicine （TCM） possesses the unique advantages of a holistic perspective and syndrome differentiation-based treatment， achieving breakthrough progress in targeted PCD intervention for ALI. It can mitigate pulmonary tissue injury by suppressing inflammatory responses， regulating immune function， and improving coagulation disorders， thereby delaying the onset and progression of ALI. This paper delves into the molecular mechanisms of PCD in ALI， systematically reviewing the effects of TCM extracts， active constituents （terpenoids， saponins， flavonoids， and phenols）， and compound formulas （e.g.， Qingjie Huagong prescription， Maxing Kugan Tang， and Yifei Jianpi prescription） in improving pulmonary epithelial cell function and mitigating lung injury by modulating PCD pathways. This review aims to provide theoretical underpinnings and novel directions for drug development in the clinical prevention and treatment of ALI.

Objective:To investigate the effect of the sizes of osteon-like concentric microgroove structures on the osteoclastic differentiation of macrophages on titanium surfaces, and to provide reference for the surface modification of implants.Methods:The silicon wafers sputtered with titanium were selected as the control group (smooth surface specimens) and four concentric groups (concentric circles with the maximum diameter of 200 μm, the minimum diameter of 20 μ m, the spacing of concentric circles of 10 or 30 μm, the width of microgrooves of 10 or 30 μm, and the depth of microgrooves of 5 or 10 μm) specimens (the total sample size in each group was 27). The width of microgrooves of C10-5 and C10-10 groups was 10 μm, the depth was 5 and 10 μm, and the width of microgrooves of C30-5 and C30-10 groups was 30 μ m, the depth was 5 and 10 μ m, respectively. The physicochemical properties of the material surfaces were characterized using scanning electron microscopy and contact-angle measurement. The proliferation, adhesion of macrophage-like cell line RAW264.7 and the formation of osteoclast actin-rings on the specimen surfaces were observed by cell counting kit-8 (CCK-8), immunofluorescence staining and laser confocal microscopy. Tartrate resistant acid phosphatase (TRAP) quantitative detection, real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting were used to investigate the regulation of osteon-like concentric microgroove structures on the specimen surfaces on the osteoclastic differentiation of macrophages.Results:Macrophages aggregated and grew disorderly on the surface of the smooth group, and arranged in concentric circles along the microgroove structures on the surfaces of the concentric groups. After 5 days of culture, the cell proliferation of C30 groups (the A values of C30-5 group and C30-10 group were 1.335±0.018 and 1.340±0.033, respectively) was significantly higher than that of C10 groups (the A values of C10-5 group and C10-10 group were 0.967±0.015 and 1.182±0.020, respectively)(all P<0.05). The cell proliferation of the four concentric groups was significantly higher than that of the control group (the A value was 0.796±0.012), with statistical significance (all P<0.05). After osteoclastic induction for 5 days the osteoclasts induced in the C10-5 and C10-10 groups exhibited smaller actin rings and fewer numbers. The TRAP activity in each concentric group was significantly lower than that in the control group ( P<0.05). The expression levels of osteoclast differentiation-related genes TRAP (0.610±0.022) in the C10-10 group was lowest, and CtsK (0.489±0.136, 0.445±0.037) in the C10-5 and C10-10 groups were lower compared to the smooth group and other concentric groups, with statistical significance (all P<0.05), the expression levels of osteoclast differentiation-related proteins TRAP (0.648±0.041), MMP-9 (0.688±0.026) in the C10-10 group were lowest, and CtsK (0.491±0.016, 0.453±0.010) in the C10-10 and C30-10 groups were also lower compared to the smooth group and other concentric groups, with statistical significance (all P<0.05). Conclusions:The osteon-like concentric microgroove structures inhibit the osteoclastic differentiation of macrophage-like cell line RAW264.7, with the microgrooves 10 μm wide and 10 μm deep showing the most significant inhibitory effect on the osteoclastic differentiation.

Lipids play an important role in the regulation of cell life processes.Although there are various lipid detection methods,Raman spectroscopy,a non-invasive technique,provides the detailed chemical composition of lipid profiles without a complex sample preparation procedure and possesses greater potential in basic biology,clinical diagnosis and disease therapy.In this review,we summarized the characteristics and advantages of Raman-based techniques and their primary contribution to illustrating cellular lipid metabolism.

Objective To compare the consistency of voice parameters collected by commonly used smart-phone brands in China and professional recording equipment,and to study whether smartphones can be used for voice research.Methods A total of 67 normal subjects were selected for voice recording using six different smart-phone brands(via the"Active Health"screening APP from the National Key Research and Development Program)and professional recording equipment.Acoustic voice parameters such as fundamental frequency parameters,fre-quency variation parameters,amplitude variation parameters,formant parameters,and energy parameters were ex-tracted from the vowels/a/,/i/,and/u/.A one-way ANOVA test and Tukey's HSD post-hoc comparisons were conducted on the independent variables.Results There were no significant differences between smartphones and professional recording equipment in terms of fundamental frequency parameters such as median F0,mean F0,max F0 and min F0;frequency parameters such as jitter local,jitter local absolute,jitter rap,jitter ppq5,and jitter ddp;amplitude parameters such as shimmer local,shimmer local dB,shimmer apq3,shimmer apq5,shimmer apq11,and shimmer dda;and formant parameters such as F1,F2,F3,and F4.However,significant differences were found in energy parameters such as mean energy(F=31.171,P＜0.001),max energy(F=34.193,P＜0.001),and min energy(F=5.453,P＜0.001)between smartphones and professional recording equipment.Conclusion The smartphones using the"Active Health"screening app from the National Key Research and Development Program can replace professional recording equipment for voice research.However,caution should be exercised when selec-ting energy-related acoustic parameters.

Epidermal growth factor receptor inhibitor (EGFRI) -related paronychia is a condition clearly related to EGFRI therapy, characterized by periungual erythema, edema, purulent exudates, periungual or subungual granulomatous lesions, and sometimes accompanied by thinning, fragility or even splitting and seperation of nail plates. Inhibition of epidermal function, inflammation and secondary infections, as well as angiogenesis are the core processes in the occurrence and development of EGFRI-related paronychia. This review summarizes epidemiology, pathogenesis, clinical manifestations, prevention and treatment of EGFRI-related paronychia.

Lacking therapeutic targets highlights the crucial roles of chemotherapy and radiotherapy in the clinical management of triple-negative breast cancer (TNBC). To relieve the side effects of the chemoradiotherapy combination regimen, we design and develop a self-assembled micelle nanosystem consisting of perfluorocarbon chain-modified cisplatin prodrug. By incorporating perfluorodecalin, this nanosystem can effectively carry ozone and promote irradiation-derived reactive oxygen species (ROS) production. By leveraging the perfluorocarbon sidechain, the nanosystem exhibits efficient internalization by TNBC cells and effectively escapes from lysosomal entrapment. Under X-ray irradiation, ozone-generated ROS disrupts the intracellular redox balance, thereby facilitating the release of cisplatin in a reduction-responsive manner mediated by reduced glutathione. Moreover, oxygen derived from ozone decomposition enhances the efficacy of radiotherapy by alleviating tumor hypoxia. Notably, the combination of irradiation with ozone-loaded cisplatin prodrug nano system synergistically prompts antitumor efficacy and reduces cellular/systemic toxicity in vitro and in vivo. Furthermore, the combo regimen remodels the tumor microenvironment into an immune-favored state by triggering immunogenic cell death and relieving hypoxia, which provides a promising foundation for a combination regimen of immunotherapy. In conclusion, our nanosystem presents a novel strategy for integrating chemotherapy and radiotherapy to optimize the efficacy and safety of TNBC clinical treatment.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by persistent inflammation and joint damage, accompanied by the accumulation of plasma cells, which contributes to its pathogenesis. Understanding the genetic alterations occurring during plasma cell differentiation in RA can deepen our comprehension of its pathogenesis and guide the development of targeted therapeutic interventions. Here, our study elucidates the intricate molecular mechanisms underlying plasma cell differentiation by demonstrating that PRDX1 interacts with DOK3 and modulates its degradation by the autophagy-lysosome pathway. This interaction results in the inhibition of plasma cell differentiation, thereby alleviating the progression of collagen-induced arthritis. Additionally, our investigation identifies Salvianolic acid B (SAB) as a potent small molecular glue-like compound that enhances the interaction between PRDX1 and DOK3, consequently impeding the progression of collagen-induced arthritis by inhibiting plasma cell differentiation. Collectively, these findings underscore the therapeutic potential of developing chemical stabilizers for the PRDX1-DOK3 complex in suppressing plasma cell differentiation for RA treatment and establish a theoretical basis for targeting PRDX1-protein interactions as specific therapeutic targets in various diseases.

High mobility group box 1 (HMGB1), when released extracellularly, plays a pivotal role in the development of spinal cord synapses and exacerbates autoimmune diseases within the central nervous system. In experimental autoimmune encephalomyelitis (EAE), a condition that models multiple sclerosis, the levels of extracellular HMGB1 and interleukin-33 (IL-33) have been found to be inversely correlated. However, the mechanism by which IL-33 deficiency enhances HMGB1 release during EAE remains elusive. Our study elucidates a potential signaling pathway whereby the absence of IL-33 leads to increased binding of P300/CBP-associated factor with HMGB1 in the nuclei of astrocytes, upregulating HMGB1 acetylation and promoting its release from astrocyte nuclei in the spinal cord of EAE mice. Conversely, the addition of IL-33 counteracts the TNF-α-induced increase in HMGB1 and acetylated HMGB1 levels in primary astrocytes. These findings underscore the potential of IL-33-associated signaling pathways as a therapeutic target for EAE treatment.
Animals ; Encephalomyelitis, Autoimmune, Experimental/metabolism* ; Astrocytes/metabolism* ; Interleukin-33/metabolism* ; HMGB1 Protein/metabolism* ; Acetylation ; Mice, Knockout ; Mice, Inbred C57BL ; p300-CBP Transcription Factors/metabolism* ; Mice ; Spinal Cord/metabolism* ; Cells, Cultured ; Female ; Signal Transduction