Background: Advances in intensive care have markedly improved survival after severe brain injury, leading to a growing population of patients with prolonged disorders of consciousness (pDOC). Current management of pDOC remains largely supportive, and evidence-based neuromodulatory interventions are limited; moreover, existing guidelines provide insufficiently explicit recommendations regarding mechanisms of action and objective biomarkers of treatment response. Transcutaneous auricular vagus nerve stimulation (taVNS) has emerged as a potential noninvasive intervention; however, its modulatory effects on brain function in pDOC are not yet well characterized, and the paucity of integrative mechanistic evidence has constrained its translation into routine clinical practice. Objectives: Within a multimodal assessment framework, this study aims to systematically elucidate the neurobiological mechanisms by which taVNS modulates brain function and autonomic activity in patients with pDOC, and to evaluate its clinical potential to enhance levels of consciousness. Methods: In this randomized controlled trial, 60 patients with vegetative state/minimally conscious state will be enrolled and randomly allocated to a taVNS group, a transcutaneous nonauricular vagus nerve stimulation group (sham), or a control group (n = 20 per group) for a 4-week intervention. The primary outcome will be changes in the Coma Recovery Scale-Revised scores from baseline to weeks 1, 2, and 4 of treatment. Secondary outcomes will include functional brain activity assessed by electroencephalography and functional near-infrared spectroscopy, as well as autonomic modulation indexed by heart rate variability. Functional prognosis will be evaluated using the Glasgow Outcome Scale-Extended at the end of treatment and at a 6-month follow-up. Safety will be assessed by continuous monitoring and documentation of adverse events throughout the study period. Results and discussion: By integrating electroencephalography–functional near-infrared spectroscopy with heart rate variability, this study will characterize the effects of taVNS on functional brain networks and consciousness recovery in pDOC across complementary behavioral, electrophysiological, hemodynamic, and autonomic domains, while interrogating potential sources of clinical and neurobiological heterogeneity. The findings are expected to provide a mechanistic and evidence-based foundation for the mechanism-driven clinical implementation of taVNS and the optimization of stimulation protocols in pDOC. Clinical trial registration: International Traditional Medicine Clinical Trial Registry, ITMCTR20250021041, https://itmctr.ccebtcm.org.cn.

Iron metabolism is a critical factor in tumorigenesis and development. Although TP53 mutations are prevalent in glioblastoma (GBM), the mechanisms by which TP53 regulates iron metabolism remain elusive. We reveal an imbalance iron homeostasis in GBM via TCGA database analysis. TP53 mutations disrupted iron homeostasis in GBM, characterized by elevated total iron levels and reduced ferritin (FTH). The gain-of-function effect triggered by TP53 mutations upregulates itchy E3 ubiquitin-protein ligase (ITCH) protein expression in astrocytes, leading to FTH degradation and an increase in free iron levels. TP53-mut astrocytes were more tolerant to the high iron environment induced by exogenous ferric ammonium citrate (FAC), but the increase in intracellular free iron made them more sensitive to Erastin-induced ferroptosis. Interestingly, we found that Erastin combined with FAC treatment significantly increased ferroptosis. These findings provide new insights for drug development and therapeutic modalities for GBM patients with TP53 mutations from iron metabolism perspectives.
Ferroptosis/drug effects* ; Humans ; Iron/metabolism* ; Glioblastoma/metabolism* ; Tumor Suppressor Protein p53/metabolism* ; Homeostasis/physiology* ; Ferritins/metabolism* ; Brain Neoplasms/genetics* ; Mutation ; Astrocytes/drug effects* ; Cell Line, Tumor ; Piperazines/pharmacology* ; Quaternary Ammonium Compounds/pharmacology* ; Ferric Compounds

ObjectiveTo investigate the diagnostic accuracy and grading capability of ultrasound-derived fat fraction （UDFF）， controlled attenuation parameter （CAP）， and hepatic/renal ratio （HRR） in assessing hepatic steatosis in metabolic associated fatty liver disease （MAFLD） with magnetic resonance imaging-proton density fat fraction （MRI-PDFF） as the gold standard. MethodsA total of 150 patients with MAFLD who attended The First Hospital of Hebei Medical University from January 2023 to December 2024 were enrolled， and 148 healthy volunteers were recruited. All subjects underwent MRI-PDFF， UDFF， CAP， and HRR examinations. Hepatic steatosis was graded based on MRI-PDFF （S0：148 cases； S1：92 cases； S2：21 cases； S3：37 cases）， and the MAFLD patients with different grades of hepatic steatosis were compared in terms of UDFF， CAP， HRR， and clinical features. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups and the Tukey HSD test was used for further comparision between two groups； the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups， and the Mann-Whitney U test was used for further comparison between two groups； the chi-square test was used for comparison of categorical data between groups. The Spearman correlation analysis was used to investigate the correlation between UDFF， CAP， HRR， and MRI-PDFF in different grades of MAFLD； the receiver operating characteristic （ROC） curve was used to investigate the efficacy of UDFF， CAP， and HRR in the diagnosis of different degrees of hepatic steatosis in MAFLD； the Bland-Altman difference plot was used to analyze the consistency between UDFF and MRI-PDFF in different degrees of hepatic steatosis in MAFLD. ResultsUDFF measurement gradually increased with the increase in the grade of fatty liver （H=201.52，P0.001）. The Spearman correlation analysis showed that there was a strong correlation between any two indicators of UDFF， CAP， HRR， and MRI-PDFF in S1， S2， and S3 MAFLD （all P0.001）， with the strongest correlation between UDFF and MRI-PDFF （r_s1=0.884，r_s2=0.962，r_s3=0.929， all P0.001）. The ROC curve analysis showed that UDFF had a larger area under the ROC curve （AUC） than CAP and HRR in the graded diagnosis of S1 and S3 （all P0.05）， while in the diagnosis of S2 MAFLD， UDFF had a significantly larger AUC than HRR （P0.05） and a similar AUC to CAP （P0.05）. The Bland-Altman difference plot showed good consistency between UDFF and MRI-PDFF in different degrees of hepatic steatosis in MAFLD. ConclusionCompared with CAP and HRR， UDFF can accurately measure liver fat content and has good efficacy in identifying varying degrees of hepatic steatosis in MAFLD.

Objective:To explore the effect of triple pre-rehabilitation strategy on improving nutritional status, negative emotions and quality of life in breast cancer patients undergoing neoadjuvant chemotherapy, so as to provide reference for clinical practice.Methods:A quasi experimental research was adopted. A total of 133 patients who underwent neoadjuvant chemotherapy for breast cancer at Gansu Provincial Cancer Hospital between December 2023 and December 2024 were recruited as research participants. Patients were stratified into the experimental group ( n = 66) and control group ( n = 67) according to their admission sequence. The control group received routine health guidance, while the experimental group was provided with a triple pre-rehabilitation strategy based on the control group. The serum albumin, body mass index, anxiety, depression scores and quality of life scores were compared between the two groups before intervention, 3 weeks after intervention and 2 weeks post-surgery. Results:Totally 64 patients in each group completed the study. The age of the experimental group was (49.50 ± 10.46) years, and that of the control group was (50.42 ± 12.50) years. All of them were females. After a 3-week intervention, the serum albumin levels in the experimental group were (42.62 ± 1.56) g/L, compared to (40.50 ± 1.31) g/L 2 weeks post-surgery. In the control group, these values were (40.98 ± 1.35) g/L and (40.00 ± 1.67) g/L, respectively. The differences between groups were statistically significant ( t = -6.38, -1.89, both P<0.05). Additionally, body mass index of the experimental group was (24.02 ± 1.35) kg/m 2 after the 3-week intervention and (23.48 ± 1.72) kg/m 2 2 weeks post-surgery, which were significantly higher than those of the control group (22.98 ± 1.72) kg/m 2 and (22.50 ± 1.31) kg/m 2, respectively ( t = - 3.81, - 3.61, both P<0.05). The anxiety scores in the experimental group were (6.00 ± 2.09) of 3 weeks after intervention and (7.00 ± 2.62) 2 weeks post-surgery, both of which were significantly lower than those in the control group (7.00 ± 2.26) and (8.00 ± 2.76), with statistically significant differences ( t = 2.60, 2.10, both P<0.05). Additionally, the depression scores in the experimental group were (5.00 ± 2.20) of 3 weeks after intervention and (6.00 ± 2.40) 2 weeks post-surgery, which were also significantly lower than those in the control group (6.00 ± 2.02) and (7.00 ± 2.09), with statistically significant differences ( t = 2.68, 2.52, both P<0.05). Furthermore, the quality of life scores in the experimental group were (94.84 ± 18.00) of 3 weeks after intervention and (84.02 ± 17.13) 2 weeks post-surgery, which were significantly higher than those in the control group (82.00 ± 18.75) and (72.03 ± 17.10), with statistically significant differences ( t = - 3.95, - 3.96, both P<0.05). Conclusions:The triple pre-rehabilitation strategy plays a crucial role in mitigating the nutritional risk for breast cancer patients undergoing neoadjuvant chemotherapy, alleviating their negative emotions, and enhancing their quality of life. This approach is anticipated to serve as a valuable reference for preoperative health guidance tailored to breast cancer patients undergoing neoadjuvant chemotherapy.

Objective To observe the value of machine learning(ML)models based on quantitative ultrasound(QUS)and clinical indexes for predicting metabolic associated fatty liver disease(MAFLD).Methods Totally 298 patients underwent abdominal MR and QUS examinations were retrospectively enrolled,including 150 cases with and 148 cases without MAFLD.The patients were divided into training set(including 107 cases of MAFLD and 101 cases of non-MAFLD)and test set(including 43 cases of MAFLD and 47 cases of non-MAFLD)at a ratio of 7∶3.Features were selected using least absolute shrinkage and selection operator(LASSO)regression and logistic regression(LR),based on which predictive models were constructed using 6 ML classifiers,including Gaussian naive Bayes(GNB),LR,random forest(RF),support vector machine(SVM),extreme gradient boosting(XGBoost)and K-nearest neighbor(KNN),respectively.Then the receiver operating characteristic curves were drawn,and the area under the curve(AUC)and the Brier score were calculated to evaluate the predictive efficacy of the models.Results The elevated age,glutamic-pyruvic transaminase(GPT),glutamic-oxaloacetic transaminase(GOT),uric acid(UA),low-density lipoprotein cholesterol(LDL-C),controlled attenuation parameter(CAP),ultrasound-derived fat fraction(UDFF)and shear wave velocity(SWV),as well as blurred liver contour were all independent indicators for higher likelihood of MAFLD(all P＜0.05).The AUC and Brier score of XGBoost model in training set was 0.991 and 0.006,in test set was 0.973 and 0.069,respectively,both higher than those of other models,and decision curve analysis(DCA)indicated that XGBoost model had high net benefit.Conclusion ML models based on QUS and clinical indexes,especially XGBoost model had high efficacy for predicting MAFLD.

OBJECTIVE To explore the mechanism of action of Qihuang Zhuyu formula(QHZYF)in improving depression after myocardial infarction(MI),with a focus on revealing its regulatory effect on the inflammatory response of the heart and brain.METH-ODS The active ingredients of QHZYF and the action targets for intervening in depression after MI were analyzed by using ultra-per-formance liquid chromatography-high-resolution mass spectrometry(UPLC-Q-TOF/MS)combined with network pharmacology and molecular docking.A rat model of depression after MI was established by ligation of the left anterior descending coronary artery com-bined with chronic restraint stress.Echocardiography was used to evaluate cardiac function,hematoxylin-eosin(HE)and Masson stai-ning were used to evaluate myocardial injury,behavioral tests were used to detect melancholic behaviors,Nissl staining was used to e-valuate hippocampal neuron injury.Western blot detection of tumor necrosis factor receptor 2(TNFR2),phosphatidylinositol-3-kinase(PI3K),phosphorylated seronine protein kinase(p-AKT),seronine protein kinase(AKT),tumor necrosis factor receptor 1(TNFR1),phosphorylated nuclear factor κB(p-NF-κB),and nuclear factor κB(NF-κB)in cardiac and hippocampal tissues was conducted.The levels of serum IL-6 and IL-10 were detected by enzyme-linked immunosorbent assay(ELISA),and the expression of TNFR1 and TNFR2 was detected by immunohistochemical technique(IHC).In vitro experiments,co-culture of rat cardiomyocyte line H9C2 cells and rat adrenal pheochromocytoma cell line with high differentiation PC12 cells was conducted,TNFR1 inhibitor(H398)and TNFR2 agonist(C-6His)were administered for intervention,and the expression of TNFR2,PI3K,p-AKT,AKT,TN-FR1,NF-κB,p-NF-κB was detected by Western blot.Observe the apoptosis of cells by TUNEL staining,ELISA was used to detect the levels of IL-6 and IL-10 in the cell supernatant.RESULTS Network pharmacological analysis indicates that the TNF signaling pathway was a key target for the treatment of depression after MI with the QHZYF.In vivo experiments have confirmed that the interven-tion of QHZYF could significantly improve the cardiac function,myocardial tissue and hippocampal neuron structure damage of de-pressed rats after MI,and improve their depression-like behaviors.At the molecular level,the high-dose group of QHZYF significant-ly upregulated TNFR2,p-AKT/AKT,and IL-10 in cardiac and hippocampal tissues(P＜0.01),and downregulated TNFR1,p-NF-κB/NF-κB and IL-6(P＜0.01).In vitro experiments showed that the drug-containing serum of QHZYF significantly upregulated the expression of TNFR2,p-AKT/AKT and IL-10 in H9C2 and PC12 cells(P＜0.01),downregulated the expression of TNFR1,p-NF-κB/NF-κB and IL-6(P＜0.01),and significantly inhibited cell apoptosis(P＜0.01).Furthermore,experiments on the combined ap-plication of H398 or C-6His further confirmed that its protective and anti-inflammatory effects were mediated by regulating the TN-FR2/PI3K/AKT and TNFR1/NF-κB pathways.CONCLUSION QHZYF improves the homeostasis of heart and brain inflammation by regulating the TNF pathway,and ameliorates myocardial injury and depressive state in depressed rats after MI.

OBJECTIVE To explore the mechanism of action of Qihuang Zhuyu formula(QHZYF)in improving depression after myocardial infarction(MI),with a focus on revealing its regulatory effect on the inflammatory response of the heart and brain.METH-ODS The active ingredients of QHZYF and the action targets for intervening in depression after MI were analyzed by using ultra-per-formance liquid chromatography-high-resolution mass spectrometry(UPLC-Q-TOF/MS)combined with network pharmacology and molecular docking.A rat model of depression after MI was established by ligation of the left anterior descending coronary artery com-bined with chronic restraint stress.Echocardiography was used to evaluate cardiac function,hematoxylin-eosin(HE)and Masson stai-ning were used to evaluate myocardial injury,behavioral tests were used to detect melancholic behaviors,Nissl staining was used to e-valuate hippocampal neuron injury.Western blot detection of tumor necrosis factor receptor 2(TNFR2),phosphatidylinositol-3-kinase(PI3K),phosphorylated seronine protein kinase(p-AKT),seronine protein kinase(AKT),tumor necrosis factor receptor 1(TNFR1),phosphorylated nuclear factor κB(p-NF-κB),and nuclear factor κB(NF-κB)in cardiac and hippocampal tissues was conducted.The levels of serum IL-6 and IL-10 were detected by enzyme-linked immunosorbent assay(ELISA),and the expression of TNFR1 and TNFR2 was detected by immunohistochemical technique(IHC).In vitro experiments,co-culture of rat cardiomyocyte line H9C2 cells and rat adrenal pheochromocytoma cell line with high differentiation PC12 cells was conducted,TNFR1 inhibitor(H398)and TNFR2 agonist(C-6His)were administered for intervention,and the expression of TNFR2,PI3K,p-AKT,AKT,TN-FR1,NF-κB,p-NF-κB was detected by Western blot.Observe the apoptosis of cells by TUNEL staining,ELISA was used to detect the levels of IL-6 and IL-10 in the cell supernatant.RESULTS Network pharmacological analysis indicates that the TNF signaling pathway was a key target for the treatment of depression after MI with the QHZYF.In vivo experiments have confirmed that the interven-tion of QHZYF could significantly improve the cardiac function,myocardial tissue and hippocampal neuron structure damage of de-pressed rats after MI,and improve their depression-like behaviors.At the molecular level,the high-dose group of QHZYF significant-ly upregulated TNFR2,p-AKT/AKT,and IL-10 in cardiac and hippocampal tissues(P＜0.01),and downregulated TNFR1,p-NF-κB/NF-κB and IL-6(P＜0.01).In vitro experiments showed that the drug-containing serum of QHZYF significantly upregulated the expression of TNFR2,p-AKT/AKT and IL-10 in H9C2 and PC12 cells(P＜0.01),downregulated the expression of TNFR1,p-NF-κB/NF-κB and IL-6(P＜0.01),and significantly inhibited cell apoptosis(P＜0.01).Furthermore,experiments on the combined ap-plication of H398 or C-6His further confirmed that its protective and anti-inflammatory effects were mediated by regulating the TN-FR2/PI3K/AKT and TNFR1/NF-κB pathways.CONCLUSION QHZYF improves the homeostasis of heart and brain inflammation by regulating the TNF pathway,and ameliorates myocardial injury and depressive state in depressed rats after MI.

Objective To observe the value of machine learning(ML)models based on quantitative ultrasound(QUS)and clinical indexes for predicting metabolic associated fatty liver disease(MAFLD).Methods Totally 298 patients underwent abdominal MR and QUS examinations were retrospectively enrolled,including 150 cases with and 148 cases without MAFLD.The patients were divided into training set(including 107 cases of MAFLD and 101 cases of non-MAFLD)and test set(including 43 cases of MAFLD and 47 cases of non-MAFLD)at a ratio of 7∶3.Features were selected using least absolute shrinkage and selection operator(LASSO)regression and logistic regression(LR),based on which predictive models were constructed using 6 ML classifiers,including Gaussian naive Bayes(GNB),LR,random forest(RF),support vector machine(SVM),extreme gradient boosting(XGBoost)and K-nearest neighbor(KNN),respectively.Then the receiver operating characteristic curves were drawn,and the area under the curve(AUC)and the Brier score were calculated to evaluate the predictive efficacy of the models.Results The elevated age,glutamic-pyruvic transaminase(GPT),glutamic-oxaloacetic transaminase(GOT),uric acid(UA),low-density lipoprotein cholesterol(LDL-C),controlled attenuation parameter(CAP),ultrasound-derived fat fraction(UDFF)and shear wave velocity(SWV),as well as blurred liver contour were all independent indicators for higher likelihood of MAFLD(all P＜0.05).The AUC and Brier score of XGBoost model in training set was 0.991 and 0.006,in test set was 0.973 and 0.069,respectively,both higher than those of other models,and decision curve analysis(DCA)indicated that XGBoost model had high net benefit.Conclusion ML models based on QUS and clinical indexes,especially XGBoost model had high efficacy for predicting MAFLD.

Objective:To explore the effect of triple pre-rehabilitation strategy on improving nutritional status, negative emotions and quality of life in breast cancer patients undergoing neoadjuvant chemotherapy, so as to provide reference for clinical practice.Methods:A quasi experimental research was adopted. A total of 133 patients who underwent neoadjuvant chemotherapy for breast cancer at Gansu Provincial Cancer Hospital between December 2023 and December 2024 were recruited as research participants. Patients were stratified into the experimental group ( n = 66) and control group ( n = 67) according to their admission sequence. The control group received routine health guidance, while the experimental group was provided with a triple pre-rehabilitation strategy based on the control group. The serum albumin, body mass index, anxiety, depression scores and quality of life scores were compared between the two groups before intervention, 3 weeks after intervention and 2 weeks post-surgery. Results:Totally 64 patients in each group completed the study. The age of the experimental group was (49.50 ± 10.46) years, and that of the control group was (50.42 ± 12.50) years. All of them were females. After a 3-week intervention, the serum albumin levels in the experimental group were (42.62 ± 1.56) g/L, compared to (40.50 ± 1.31) g/L 2 weeks post-surgery. In the control group, these values were (40.98 ± 1.35) g/L and (40.00 ± 1.67) g/L, respectively. The differences between groups were statistically significant ( t = -6.38, -1.89, both P<0.05). Additionally, body mass index of the experimental group was (24.02 ± 1.35) kg/m 2 after the 3-week intervention and (23.48 ± 1.72) kg/m 2 2 weeks post-surgery, which were significantly higher than those of the control group (22.98 ± 1.72) kg/m 2 and (22.50 ± 1.31) kg/m 2, respectively ( t = - 3.81, - 3.61, both P<0.05). The anxiety scores in the experimental group were (6.00 ± 2.09) of 3 weeks after intervention and (7.00 ± 2.62) 2 weeks post-surgery, both of which were significantly lower than those in the control group (7.00 ± 2.26) and (8.00 ± 2.76), with statistically significant differences ( t = 2.60, 2.10, both P<0.05). Additionally, the depression scores in the experimental group were (5.00 ± 2.20) of 3 weeks after intervention and (6.00 ± 2.40) 2 weeks post-surgery, which were also significantly lower than those in the control group (6.00 ± 2.02) and (7.00 ± 2.09), with statistically significant differences ( t = 2.68, 2.52, both P<0.05). Furthermore, the quality of life scores in the experimental group were (94.84 ± 18.00) of 3 weeks after intervention and (84.02 ± 17.13) 2 weeks post-surgery, which were significantly higher than those in the control group (82.00 ± 18.75) and (72.03 ± 17.10), with statistically significant differences ( t = - 3.95, - 3.96, both P<0.05). Conclusions:The triple pre-rehabilitation strategy plays a crucial role in mitigating the nutritional risk for breast cancer patients undergoing neoadjuvant chemotherapy, alleviating their negative emotions, and enhancing their quality of life. This approach is anticipated to serve as a valuable reference for preoperative health guidance tailored to breast cancer patients undergoing neoadjuvant chemotherapy.

It is difficult to accurately distinguish pharyngitis caused by Group A Streptococcus(GAS) from other pathogens according to the clinical presentation alone, which cannot effectively guide the rational use of antimicrobials.The pharyngeal swab culture, rapid antigen detection test, nucleic acid test, and blood test can help definitively diagnose GAS pharyngitis.However, there are differences in different guidelines on who the laboratory test methods are intended for, interpretation of laboratory test results and so on.This article summarizes and analyses the laboratory diagnostic modalities and their characteristics, as well as recommendations for GAS pharyngitis in different guidelines to provide references for the clinical diagnosis, antimicrobial treatment, and further study of GAS pharyngitis.