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.

OBJECTIVE To investigate the improvement effect and mechanism of processed Oxytropis falcata on renal fibrosis （RF） in rats. METHODS RF model was induced by adenine. After modeling， the rats were divided into the model group， positive control group （colchicine， 0.45 mg/kg）， and processed O. falcata low-， medium- and high-dose groups （0.5， 1， 2 g/kg）， respectively. Additionally， a blank group without modeling was set up， with 8 rats in each group. The positive control group and the various dosage groups of processed O. falcata were given the corresponding medicinal solutions intragastrically， while the blank group and model group were given equal volume of normal saline intragastrically， once daily for 28 consecutive days. The appearance and histopathological morphology of the rats’ kidneys were observed. Serum levels of renal function indexes [bl ood urea nitrogen （BUN）， creatinine （Cr） ] and inflammatory factors [interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） ] in rats were detected. Protein expressions of fibronectin （FN）， α -smooth muscle actin （ α -SMA） and collagen type Ⅰ （Col-Ⅰ） in renal tissue of rats were determined. mRNA expressions of transforming growth factor-β 1 （TGF-β 1 ）， Smad3 and extracellular signal-regulated kinase 1/2 （ERK1/2） in renal tissues were measured. Protein expression of TGF-β 1 and phosphorylation levels of Smad3 and ERK1/2 in renal tissues were detected. RESULTS Compared with the blank group， the rats in the model group exhibited enlarged kidneys with pale color， rough and uneven surface. There was a significant infiltration of inflammatory cells and vacuolated cells in the renal tubules， along with marked proliferation of collagen fibers. Serum levels of BUN， Cr， IL-6 and TNF-α， protein expressions of α -SMA， Col-Ⅰ and FN in renal tissues， mRNA expressions of TGF-β 1 ， Smad3， ERK1 and ERK2 and protein expression of TGF-β 1 as well as phosphorylation levels of Smad3 and ERK1/2 in renal tissues were increased significantly （ P ＜0.05）. Compared with the model group， renal pathological changes of rats were alleviated in processed O. falcata groups， with reduced infiltration of inflammatory cells and proliferation of collagen fibers. The levels of the aforementioned quantitative indicators were all significantly reversed （ P ＜0.05）. CONCLUSIONS Processed O. falcata can improve renal function in RF rats， alleviate inflammatory responses， and reduce abnormal collagen fiber deposition. Its mechanism of action may be related to the inhibition of the activity of the TGF-β 1 /Smad signaling pathway.

BackgroundResident physicians represent a high-risk group for sleep disorders， exhibiting a significantly higher prevalence of such conditions compared with the general population， which severely impairs their physical and mental health. It is hypothesized that perceived stress negatively impacts sleep quality through psychological mechanisms， such as depleting self-control resources and triggering anxiety. However， this pathway warrants empirical validation. ObjectiveTo explore the mediating role of self-control and anxiety emotions in the association between perceived stress and sleep quality among resident physicians， and to elucidate the underlying psychological mechanisms， aiming at providing theoretical basis for developing targeted psychological interventions. MethodsA cross-sectional survey was conducted in April 2025. First- to third- year resident physicians at a hospital in Fuyang City were recruited as participants （n=372）. The Chinese Perceived Stress Scales （CPSS）， the Chinese version of the Dual-Mode of Self-Control Scale （DMSC-S）， the Pittsburgh Sleep Quality Index （PSQI）， and the Generalized Anxiety Disorder Scale-7 item （GAD-7） were used for group testing. The model 6 of the Process macro version 4.1 was ultilized to examine the mediating pathway of self-control and anxiety emotions between perceived stress and sleep quality. ResultsA total of 322 valid questionnaires were collected， yielding an effective responsive rate of 86.56%. Among the respondents， 146 （45.34%） reported poor sleep quality. The CPSS score and GAD-7 score of resident physicians were positively correlated with the PSQI score （r=0.727， 0.784， P<0.01）， while the DMSC-S score was negatively correlated with the PSQI score （r=-0.615， P<0.01）. Perceived stress directly and positively predicted poor sleep quality （B=0.124， P<0.01）， with the direct effect accounting for 31.39% of the total effect. Furthermore， perceived stress indirectly affected sleep quality through the independent mediating effects of self-control and anxiety emotions. The indirect effect values of 0.053 （95% CI： 0.019 - 0.091） and 0.192 （95% CI： 0.141 - 0.249）， accounting for 13.42% and 48.61% of the total effect， respectively. Perceived stress also impact sleep quality through the serial mediating effect of self-control and anxiety， with the indirect effect value of 0.026 （95% CI： 0.005 - 0.049）， accounting for 6.58% of the total effect. ConclusionThe perceived stress of resident physicians can influence sleep quality by impairing self-control， exacerbating anxiety， and through the serial mediation of both factors.

ObjectiveTo investigate the potential targets of 2，5-dihydroxybenzoic acid （2，5-DHBA） in the treatment of metabolic dysfunction-associated fatty liver disease （MAFLD） and the molecular mechanism by which it regulates the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. MethodsA network pharmacology analysis was performed at first， and related databases were used to obtain the common action targets of 2，5-DHBA and MAFLD， followed by molecular docking and pathway enrichment analysis to predict the potential biological processes and signaling pathways regulated by these targets. Then mouse normal hepatocytes AML-12 were used for cell experiments， and CCK-8 assay was used to determine the three optimal intervention concentrations of 2，5-DHBA. AML-12 cells were divided into control group， model group （cells cultured in a medium containing 2 mmol/L free fatty acid ［FFA］ to establish a cell model of MAFLD）， and three 2，5-DHBA intervention groups （cells cultured with FFA and 2，5-DHBA at the three different concentrations of 1.25， 5， and 20 μmol/L， respectively）. Oil red O staining was used to observe intracellular lipid accumulation； the fluorescent probe DCFH-DA was used to measure the level of intracellular reactive oxygen species （ROS）； biochemical assays were used to measure the content of triglyceride （TG） and total cholesterol （TC）； quantitative real-time PCR and Western Blot were used to measure the expression levels of related genes and proteins. A one-way analysis of variance was used for comparison between multiple groups， and the Tukey’s test was used for further comparison between two groups. ResultsA total of 32 common targets were obtained for 2，5-DHBA and MAFLD. Within a concentration range of 1.25 — 20 μmol/L， 2，5-DHBA treatment， starting from the concentration of 2.5 μmol/L， increased the viability of AML-12 cells in a dose-dependent manner. The minimum effective concentration of 1.25 μmol/L， the intermediate gradient concentration of 5 μmol/L， and the maximum safe concentration within the effective range of 20 μmol/L were selected for low-， middle-， and high-dose intervention， respectively. Compared with the model group， 2，5-DHBA intervention at concentrations of 1.25， 5， and 20 μmol/L could significantly reduce lipid droplet accumulation in cells （all P<0.05）， and there were significant differences in TG and TC between the model group and the three intervention groups （all P<0.05）. Compared with the control group， the model group had a significant increase in ROS fluorescence intensity （P<0.05）， and compared with the model group， there was a dose-dependent reduction in intracellular ROS level after treatment with 5 μmol/L or 20 μmol/L 2，5-DHBA （P<0.05）. Compared with the model group， 2，5-DHBA treatment at concentrations of 5 and 20 μmol/L significantly downregulated the mRNA expression levels of the lipogenic genes SREBf1 and FASN and upregulated the mRNA expression levels of the key antioxidant gene Nrf2 and its downstream target gene HO-1 （all P<0.05）. Compared with the control group， the expression of p-PI3K/PI3K and p-Akt/Akt in the model group was significantly increased （all P<0.05）. Compared with the model group， the protein expression levels of p-PI3K/PI3K and p-Akt/Akt were significantly decreased after 5 μmol/L 2，5-DHBA treatment （P<0.05）. Compared with the control group， the expression levels of Nrf2 and HO-1 protein in the model group were significantly decreased （all P<0.05）. The protein levels of Nrf2 and HO-1 were significantly increased after treatment with 1.25 μmol/L 2，5-DHBA （P<0.05）. ConclusionNetwork pharmacology and cellular experiments confirm that 2，5-DHBA can alleviate lipid deposition and oxidative stress by regulating the PI3K/Akt signaling pathway， thereby exerting a therapeutic effect on MAFLD.

Objective To systematically evaluate the risk prediction models for postoperative pulmonary infection in patients with esophageal cancer, providing an objective basis for clinical selection and optimization of models. Methods A systematic search was conducted in Chinese and English databases such as VIP, Wanfang, CNKI, CBM, PubMed, Cochrane Library, Embase, and Web of Science for studies related to the risk prediction models of postoperative pulmonary infection in patients with esophageal cancer from the inception to September 30, 2024. The PROBAST tool was used to assess the quality of prognostic model research, and the RevMan 5.4 software was used for meta-analysis of predictive factors. Results A total of 17 articles were included, containing 26 pulmonary infection risk prediction models. The area under the receiver operating characteristic curve (AUC) ranged from 0.627 to 0.942, among which 21 models had good predictive performance (AUC>0.7). Quality assessment through the PROBAST tool revealed that all 17 articles had a high risk of bias. Meta-analysis results showed that common predictive factors for postoperative pulmonary infection in esophageal cancer included smoking history (OR=1.97), smoking index ≥200 (cigarettes-years) (OR=4.38), smoking index ≥400 (cigarettes-years) (OR=2.00), age (OR=1.27), comorbid diabetes (OR=2.13), comorbid emphysema or chronic obstructive pulmonary disease (OR=1.55), low plasma albumin levels (OR=1.17), prognostic nutritional index (OR=4.45), history of related lung diseases (OR=2.10), tumor location (OR=2.32), surgical approach (OR=2.21), operation time (OR=1.73), preoperative serum procalcitonin levels (OR=3.06), anastomotic leakage (OR=3.39), reduced forced expiratory volume in the first second/forced vital capacity ratio (OR=0.86), and hoarseness (OR=2.23). Conclusion At present, the risk prediction models for postoperative pulmonary infection in esophageal cancer are still in the stage of continuous development and optimization, and their research quality needs to be further improved. Future research can refer to the predictive factors summarized in this study based on meta-analysis, combined with clinical practice, to select appropriate methods to construct and validate the risk prediction models for postoperative pulmonary infection in esophageal cancer, thus providing early targeted preventive strategies for high-risk patients.

BACKGROUND:In recent years,numerous studies have shown that autophagy and mitophagy play an important role in the regulation of bone metabolism.Under non-physiological conditions,mitophagy breaks the balance of bone metabolism and triggers metabolism disorders,which affect osteoblasts,osteoclasts,osteocytes,chondrocytes,bone marrow mesenchymal stem cells,etc. OBJECTIVE:To summarize the mechanism of mitophagy in regulating bone metabolic diseases and its application in clinical treatment. METHODS:PubMed,Web of Science,CNKI,WanFang and VIP databases were searched by computer using the keywords of"mitophagy,bone metabolism,osteoblasts,osteoclasts,osteocytes,chondrocytes,bone marrow mesenchymal stem cells"in English and Chinese.The search time was from 2008 to 2023.According to the inclusion criteria,90 articles were finally included for review and analysis. RESULTS AND CONCLUSION:Mitophagy promotes the generation of osteoblasts through SIRT1,PINK1/Parkin,FOXO3 and PI3K signaling pathways,while inhibiting osteoclast function through PINK1/Parkin and SIRT1 signaling pathways.Mitophagy leads to bone loss by increasing calcium phosphate particles and tissue protein kinase K in bone tissue.Mitophagy improves the function of chondrocytes through PINK1/Parkin,PI3K/AKT/mTOR and AMPK signaling pathways.Modulation of mitophagy shows great potential in the treatment of bone diseases,but there are still some issues to be further explored,such as different stages of drug-activated mitophagy,and the regulatory mechanisms of different signaling pathways.

Childhood primary renal tubular diseases are chronic kidney diseases characterized by impaired renal tubular reabsorption. Primary renal tubular disease has diverse clinical manifestations and lacks of specificity. Laboratory tests are limited，making it prone to missed diagnosis and misdiagnosis. Based on the current knowledge of renal tubular diseases，authors propose early warning signals of renal tubular diseases such as family history of primary tubular diseases，unexplained polyhydramnios during pregnancy，polydipsia，polyuria，delayed growth and development or rickets，decreased muscle strength and tone，unexplained electrolyte disturbance，hyperuricemia，acid-base disturbance，positive urine sugar test，renal tubular proteinuria，urinary imaging examination suggesting kidney stones，calcium deposition，renal cysts and early onset of eye，ear，joint and neuron injury.Meanwhile，some universal management strategies for primary renal tubular disease are proposed，emphasizing the importance of multidisciplinary collaboration，genetic testing and individualized intervention to improve the long-term prognosis of childhood primary renal tubular diseases.

Non-small cell lung cancer (NSCLC) is a malignant tumor with a high global incidence rate, accounting for about 10.54% of all new cancer cases and posing a serious threat to human health. Due to significant individual variations in the efficacy of immunotherapy among NSCLC patients, it is necessary to identify accurate detection indicators to screen appropriate populations, monitor treatment efficacy, and assist in prognosis assessment. Programmed death-ligand 1 (PD-L1), as an immunosuppressive molecule expressed on the surface of tumor cells and various immune cell membranes, can serve as a "companion diagnostic" or "supplementary diagnostic" tool to guide clinical treatment decisions for metastatic NSCLC patients. Given that tumor tissue PD-L1 testing is an invasive procedure and its reliability is still under debate, the assessment of PD-L1 expression via liquid biopsies, such as circulating tumor cells, will play a significant role in predicting treatment response and prognosis in NSCLC patients.

Objective: To investigate the impact of preschool children s aquatic motor skill (AMS) acquisition on their fundamental motor skill (FMS) development and the correlation between AMS and FMS development, so ao to provide a scientific basis for early childhood education and physical education teaching. Methods: From April to June 2024, 60 children, recruited by random sampling from a kindergarten in Taiyuan, were stratified randomly divided into an experimental group ( n =30) and a control group ( n =30). The experimental group were further divided into five classes of six each. They received AMS practice interventions twice weekly, 40 minutes per session, over eight weeks (16 sessions total) at a designated swimming center. The control group maintained their usual routine. Children s FMS and AMS were assessed pre and post intervention using the Test of Gross Motor Development-3rd (TGMD3) and Actual Aquatic Skills Test (AAST), respectively. Before and after test comparisons within and between groups employed t-tests, Wilcoxon signedrank tests, ANCOVA (including ranktransformed ANCOVA), and Cohen s d effect sizes were calculated for standardized mean differences. Spearman rank correlation was used to analyze relationships between FMS and AMS. Results: After the aquatic learning, the experimental group scored significantly higher than the control group on locomotor skills ( F=20.47, P <0.01, η 2=0.26), FMS ( F=4.59, P =0.04, η 2=0.08), and AMS ( F=109.71, P<0.01, η 2=0.79). The experimental groups improvement in locomotor skills 5.0(3.8, 7.3) versus the control group (2.8±2.5) yielded a medium effect size (Cohen s d =0.71); AMS gains in the experimental group [26.0(20.8, 28.0)] versus controls [1.0(0, 2.3)] showed a very large effect size (Cohen s d =4.73) (both P <0.01). Among preschool children, AMS acquisition was positively correlated with locomotor skills ( r =0.39) and overall FMS ( r =0.43)(both P <0.05). Skill specific assessments revealed lower proficiency in headfirst entry (immersion), treading water, and sagittalplane rotation. Conclusion Preschool children s acquisition of AMS has a positive effect on their FMS, with mutual facilitation between the two, especially in locomotor abilities.

ObjectiveTo investigate the intervention effects and mechanisms of the flavonoid hyperoside （Hyp） on lipopolysaccharide （LPS）-induced inflammation in the zebrafish model. MethodsZebrafish larvae were either microinjected with 0.5 g·L^-1 LPS or immersed in 1 g·L^-1 LPS for the modeling of inflammation. The larvae were then treated with Hyp at 25， 50， and 100 mg·L^-1 through immersion for four consecutive days. The inflammatory phenotypes were assessed by analyzing the mortality rate， malformation rate， body length， and yolk sac area ratio. Behavioral tests were conducted to evaluate the inflammatory stress responses， and macrophage migration was observed by fluorescence microscopy. Additionally， the mRNA levels of inflammation-related genes， including interleukin-1β （IL-1β）， interleukin-6 （IL-6）， chemokine C-C motif ligand 2 （CCL2）， chemokine C-X3-C motif receptor 1 （CX3CR1）， chemokine C-C motif receptor 2 （CCR2）， and genes associated with the Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor-kappa B （NF-κB） signaling pathway， were measured by Real-time quantitative polymerase chain reaction（Real-time PCR）. ResultsCompared with the pure water injection group， the model group exhibited increased mortality， malformation rates and yolk sac area ratio （P0.01）， reduced body length （P0.01）， increased total swimming distance and high-speed swimming duration （P0.01）， and up-regulated mRNA levels of TLR4， MyD88， NF-κB， IL-1β， IL-6， CCL2， CX3CR1， and CCR2 （P0.01）. Hyp at low， medium and high doses， as well as aspirin， reduced the mortality and malformation rates （P0.05，P0.01）， increased the body length （P0.05，P0.01）， decreased the yolk sac area ratio （P0.01）， reduced the high-speed swimming duration （P0.01）， and down-regulated the mRNA levels of TLR4， MyD88， NF-κB， IL-1β， IL-6， CCL2， CX3CR1， and CCR2 （P0.05，P0.01） compared with the model group. ConclusionHyp may modulate the TLR4/MyD88/NF-κB pathway to ameliorate inflammatory phenotypes and alleviate stress conditions in zebrafish， thereby exerting the anti-inflammatory effect.