Objective: To investigate the damaging effects of red blood cell supernatant (RBC-S) stored for different durations (7 d, 14 d, and 28 d) on vascular endothelial cells, and to explore the underlying mechanisms using bioinformatics analysis, so as to provide references for optimizing red blood cell transfusion strategies. Methods: Human umbilical vein endothelial cells (HUVECs) were co-cultured with RBC-S stored for 7, 14 and 28 days, designated as the 7 d group, 14 d group and 28 d group respectively, which were collectively defined as the experimental groups. Cell damage was evaluated by cell proliferation assay (Cell Counting Kit8, CCK8), lactate dehydrogenase (LDH) release assay, 4′, 6diamidino2phenylindole (DAPI) staining, and flow cytometry for apoptosis and reactive oxygen species (ROS) levels. The damage degree of RBC-S on vascular endothelial cells was assessed by statistical analysis of damage data among different groups. Since the damage effect reached a plateau at all time points, the 28 d storage group was selected as the representative for further mechanistic studies. Transcriptomic analysis was performed to explore the role of frizzled class receptor 1 (FZD1) and Wnt signaling pathway in red blood cell storagerelated endothelial dysfunction. Results: Compared with the control group, the storage groups treated with 7 d, 14 d, and 28 d RBC-S showed significantly decreased cell proliferation rates [control group 100%, 7 d group (69.51±2.30)%, 14 d group (74.54±2.89)%, 28 d group (73.59±2.36)%, P<0.05], significantly reduced numbers of DAPI-stained cell nuclei [control group (213±12.5) per field, 7 d group (140.33±17.04) per field, 14 d group (152.00±23.72) per field, 28 d group (144.33±19.09) per field, P<0.05] and significantly increased LDH release [control group (1), 7 d group (8.33±1.41), 14 d group (9.23±0.83), 28 d group (9.16±0.60), P<0.05]. There was no significant difference in the degree of damage caused by RBC-S among different storage groups (P>0.05). With the prolongation of storage time, free hemoglobin (FHb) gradually increased [control group (not detected), 7 d (16.57±6.38) mg/L, 14 d (76.80±22.83) mg/L, 28 d (286.97±29.02) mg/L, P<0.05]. The apoptotic rate (20.53±2.94)% and ROS relative intensity (5.13±0.91) in the 28 d storage group were significantly higher than those in the control group (P<0.05). Transcriptomic analysis showed that FZD1 played a key role in vascular endothelial dysfunction induced by red blood cell storage and was closely related to the Wnt signaling regulatory network. Conclusion: RBC-S stored for 7 d, 14 d, or 28 d can all significantly damage vascular endothelial cells, and the damaging effect reaches a plateau at 7 d of storage. Mechanistic investigation of the 28 d group indicated that the downregulation of the FZD1/Wnt signaling pathway may play a critical role in vascular endothelial dysfunction induced by red blood cell storage, providing a theoretical basis for further optimizing red blood cell storage and transfusion strategies.

Background: and Purpose Vertebrobasilar fusiform aneurysms (VBFAs) carry substantial morbidity and mortality, but optimal management for unruptured VBFAs remains unclear. We compared the safety and efficacy of conservative management (CM), stent-assisted coiling (SAC), and flow diverters (FDs) in patients with unruptured VBFAs, focusing on long-term prognosis. Methods: This study included data from a nationwide Chinese cohort of patients with vertebrobasilar dissecting aneurysms. Inverse probability of treatment weighting (IPTW) balanced confounders across groups. The primary outcome was poor prognosis (modified Rankin Scale score >2). Secondary outcomes included aneurysm rupture, ischemic stroke, compression symptoms, and VBFA-related deaths. Logistic regression estimated odds ratios (ORs) and 95% confidence intervals (CIs). Subgroup and sensitivity analyses were performed. Results: Among 1,115 patients with unruptured VBFAs, 838 (median age, 54 years; 655 men) were included. After IPTW, baseline characteristics were balanced. Median follow-up was 54 months. FD was associated with a lower risk of poor prognosis than CM (OR, 0.48 [95% CI, 0.30 to 0.77]; p=0.002), with no difference between CM and SAC. FD also reduced aneurysm rupture (OR, 0.20 [95% CI, 0.07 to 0.60]; p=0.004) and compression symptoms (OR, 0.30 [95% CI, 0.13 to 0.68]; p=0.004) versus CM. Time-to-event analyses further revealed significant differences in vertebral artery lesions and Type I–II VBFAs, whereas no significant differences were observed in basilar or vertebrobasilar junction lesions or in Type III–IV VBFAs. Conclusions Compared with CM, FD was associated with improved long-term outcomes in unruptured VBFAs, particularly in vertebral artery lesions and Type I–II VBFAs, although residual confounding cannot be excluded.

Background/Aims: Natural killer (NK) cell function is generally considered dampened in chronic hepatitis B virus (HBV) infection; however, the NK cell pool exhibits phenotypic and functional heterogeneity, and the antibody--mediated effect of NK cells remains less characterized. This study evaluated the dynamic changes in antibody-mediated NK cell responses and the involvement of distinct NK subsets across disease stages and during antiviral treatment. Methods: A T-cell receptor-like antibody specific for the HBV core 18–27 peptide (cTCRL-Ab) was used to determine the antibody-mediated effect of NK cells, and an array of NK cell surface markers were analyzed in cross-sectional and longitudinal cohorts of patients with chronic HBV infection. Single-cell RNA sequencing (scRNA-seq) was performed to identify the heterogeneity of NK subsets. Results: The cTCRL-Ab enabled the detection of NK cell cytolytic activity and IFNγ production. Notably, cTCRL-Ab-mediated NK cell responses were compromised in chronically HBV-infected patients, particularly in those receiving pegylated interferon-α (Peg-IFNα), which was associated with the downregulation of CD16 expression. Correspondingly, Peg-IFNα inhibited cTCRL-Ab-mediated NK cell function by reducing CD16 expression in vitro. scRNA-seq revealed that CD16 downregulation occurred mainly within a dysfunctional CD16hi NK subset exhibiting exhaustion properties. In contrast, an activated CD16hiNK subpopulation (CX3CR1⁺KLRC2–CD16hi) with high cytotoxicity was enriched in patients who experienced favorable treatment responses. Furthermore, the intrahepatic CX3CR1+KLRC2–CD16hi subset tended to exhibit functional restoration in HBsAg-loss individuals. Conclusions Our data contribute to the understanding of antibody-mediated responses of NK cells in chronic HBV infection, and highlight a previously unappreciated functional CX3CR1+KLRC2–CD16hiNK subset as a potential therapeutic target.

As a major challenge in the global field of critical care medicine, sepsis is associated with high morbidity and mortality, driving intensive research into the development of precise risk prediction models. Among these, machine learning and deep learning-based predictive models have garnered particular attention. This article systematically reviews the evolution of sepsis risk prediction models and the challenges associated with their clinical translation, with a focus on their clinical applicability in common septic complications, including acute kidney injury, encephalopathy, and coagulation disorders. Current evidence indicates that machine learning models, by integrating multidimensional dynamic data from electronic health records, significantly enhance early warning capabilities and individualized predictive performance. However, major bottlenecks limiting clinical translation include insufficient data standardization, lack of model interpretability, and inadequate external validation. Future research should prioritize multicenter collaboration, time-series modeling strategies, and explainable artificial intelligence frameworks to optimize clinical translation pathways and facilitate the paradigm shift from traditional empirical approaches to data-driven precision strategies in sepsis risk stratification.

Metabolic dysfunction-associated fatty liver disease （MAFLD） has become the most common chronic liver disease worldwide. Its pathological process is centered on the “two-hit” theory， with gut-liver axis dysregulation running through the entire disease course from steatosis to liver cirrhosis and even hepatocellular carcinoma， and it forms an extensive cross-system regulatory mechanism with the metabolic， cardiovascular， renal， and psychological and nervous systems through multi-dimensional pathways such as the “gut-liver-brain axis”， the “liver-kidney axis”， and the “liver-heart axis”. Disease progression is not limited to the liver itself， and it also involves functional imbalance of multiple organ systems throughout the body. This article systematically elaborates on the association mechanism of “liver pathological progression-multi-axis regulation-comorbidity occurrence” in MAFLD， proposes synergistic management strategies integrating early screening based on risk stratification， targeted intervention， and multidisciplinary diagnosis and treatment， and analyzes the limitations of current research and future development directions， in order to provide a theoretical basis and practical guidance for precise diagnosis and treatment and individualized management of MAFLD.

ObjectiveTo evaluate whether the use of intrapartum ultrasound （IPUS） to monitor the labor process， as alternative method of traditional vaginal examination （VE）， could reduce the incidence of postpartum complications associated with vaginal delivery. MethodsA total of 360 primiparas who underwent cervical dilation balloon induction of labor （IOL） and achieved successful vaginal delivery after childbirth at the Third Affiliated Hospital of Sun Yat-sen University between January 2022 and December 2023， were included in the retrospective study with half assessed for labor progress using IPUS （n=180） and the other half using VE （n=180）. The differences in the relevant indicators of the labor process and the complications of vaginal delivery were compared between the two groups of pregnant women. ResultsThere was no significant difference in the duration of membrane rupture， the first stage of labor and the second stage of labor between the two groups. However， the number of VE in the IPUS group was significantly lower ［nulliparous： 4 （3-4） vs. 6 （5-8）， P＜0.001； multiparous： 2 （2-3） vs. 4 （3-5）， P＜0.001］. The IPUS group experienced a lower incidence of postpartum fever compared to the VE group ［1.67%（3/180） vs. 7.22%（13/180）， P=0.006］， while other complications of vaginal delivery， such as fever during labor， postpartum hemorrhage （PPH）， laceration， poor wound healing and uroschesis， were not significantly different （all P ＞ 0.05）. The logistic regression analysis confirmed that the use of IPUS served as a protective factor for postpartum fever with an OR of 0.06 ［95% CI （0.01， 0.36）； P=0.002］， while the duration of membrane rupture was a risk factor ［OR： 1.12； 95% CI （1.03， 1.22）； P = 0.011］. ConclusionsIPUS， as an alternative to traditional VE， does not affect the progress of labor. However， it can significantly reduce the frequency of VE and the incidence of postpartum fever， thereby facilitating the rapid recovery of postpartum women， which makes it clinically valuable for application.

ObjectiveTo investigate the association between sarcopenia and short-term prognosis in patients with acute-on-chronic liver failure （ACLF）， to assess the value of Model for End-Stage Liver Disease （MELD）， MELD combined with serum sodium concentration （MELD-Na）， MELD 3.0， and Chinese Group on the Study of Severe Hepatitis B （COSSH）-ACLF Ⅱ score in predicting the 90-day prognosis of ACLF patients， and to analyze the predictive gain of sarcopenia over traditional prognostic scoring models. MethodsA retrospective study was conducted among 219 patients with ACLF who were hospitalized in The Affiliated Hospital of Inner Mongolia Medical University， No. 2 Hospital of Hohhot， Hohhot First Hospital， and Inner Mongolia People’s Hospital from July 2018 to July 2024. L3 skeletal muscle index （L3-SMI） was measured and calculated based on abdominal computed tomography images， and sarcopenia was diagnosed based on L3-SMI. According to 90-day prognosis， the patients with ACLF were divided into survival group with 124 patients and death group with 95 patients. The chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups； the independent samples t-test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. The multivariate Cox regression analysis （stepwise backward approach） was used to identify independent risk factors for 90-day mortality in ACLF patients. The Kaplan-Meier method was used for survival analysis， and the Log-rank test was used for comparison between groups. The receiver operating characteristic curve was plotted to evaluate the predictive performance of each model， and the DeLong test， bootstrap internal validation， net reclassification improvement （NRI）， and integrated discrimination improvement （IDI） were used to assess the predictive gain of sarcopenia. ResultsThe multivariate Cox regression analysis showed that presence of sarcopenia （hazard ratio ［HR］=3.385， 95% confidence interval ［CI］： 1.722 — 6.657， P<0.05）， older age （HR=1.036， 95%CI： 1.017 — 1.056， P<0.05）， elevated alkaline phosphatase （HR=1.006， 95%CI： 1.004 — 1.009， P<0.05）， elevated total bilirubin （HR=1.003， 95%CI： 1.001 — 1.005， P<0.05）， elevated potassium （HR=1.581， 95%CI： 1.160 — 2.155， P<0.05）， elevated serum creatinine （HR=1.015， 95%CI： 1.007 — 1.024， P<0.05）， and prolonged prothrombin time （HR=1.035， 95%CI： 1.015 — 1.055， P<0.05） were independent risk factors for 90-day mortality in ACLF patients， while elevated albumin （HR=0.885， 95%CI： 0.837 — 0.935， P<0.05） was an independent protective factor against 90-day mortality. MELD， MELD-Na， MELD 3.0， and COSSH-ACLF Ⅱ scores had an area under the ROC curve （AUC） of 0.569， 0.570， 0.632， and 0.659， respectively， for predicting the 90-day prognosis of ACLF patients， suggesting that these scores had unsatisfactory performance； after they were respectively combined with sarcopenia， the AUC was increased to 0.702， 0.687， 0.723， and 0.728， respectively. The DeLong test， NRI， and IDI analyses showed the combined models had significantly better predictive performance （all P<0.05）. ConclusionSarcopenia is an independent risk factor for 90-day mortality in patients with ACLF. Traditional scoring models have limited predictive performance for 90-day prognosis in ACLF patients， and the combination with sarcopenia significantly improves predictive value， suggesting that sarcopenia can be used as an important supplementary indicator for prognostic evaluation of ACLF in clinical practice.

Lung cancer is the leading cause of cancer-related deaths worldwide. While modern medical therapies such as surgery， chemotherapy， and radiotherapy have achieved progress， they often come with severe side effects and drug resistance. Traditional Chinese medicine （TCM）， characterized by its multi-component， multi-target， and holistic regulatory properties， plays a significant role in the adjunctive treatment of lung cancer. It offers unique advantages in alleviating clinical symptoms， enhancing patients' quality of life， prolonging survival， and enhancing efficacy while reducing toxicity， serving as an important therapeutic modality for lung cancer. The theory of cancer-toxin pathogenesis proposes that phlegm-toxin is a key pathogenic factor in lung cancer， with phlegm-toxin mutual entanglement being the core pathogenesis of lung cancer. The pathogenic characteristics-viscosity， dispersion， and damage to the healthy Qi-of phlegm-toxin are closely related to the accumulation of acidic metabolic products， tumor invasion and metastasis， and immune suppression during the course of lung cancer. Phlegm resolving and toxin clearing are the core therapeutic methods for lung cancer. Such prescriptions possess dual functions of resolving phlegm and dissipating mass， as well as inhibiting tumor and detoxifying. They exert antitumor effects through multiple pathways， including regulating metabolic reprogramming of glucose and lipids， improving the acidic microenvironment of tumors， and inhibiting inflammatory signaling pathways. This paper systematically reviews the current clinical application status and mechanisms of phlegm-resolving and detoxifying Chinese herbal medicines. By integrating the theory of cancer-toxin pathogenesis with modern pharmacological research findings， this paper explores their application in lung cancer treatment and establishes a clinical application system for phlegm-resolving and detoxifying therapies in lung cancer based on phlegm-toxin pathogenesis. This system integrates ancient and modern experiences with contemporary mechanisms， aiming to provide theoretical foundations and practical references for integrated traditional and Western medicine diagnosis， treatment， and drug development for lung cancer and other conditions.

Objective To investigate the effect of total paeony glycoside(TPG)on cerebral ischemia-reperfusion injury(CI/RI)of rats.Methods The rats were randomly divided into sham surgery(sham)group,CI/RI model group(simple CI/RI group),positive control group(nimodipine group,5 mg/kg),low-dose TPG group(TPG-L group,27 mg/kg),a high-dose TPG group(TPG-H group,54 mg/kg)and a high-dose TPG+NOD-like receptor thermal protein domain associated protein 3(NLRP3)activator diethyl dithiocarbamate(DDC)group(TPG-H+DDC group,54 mg/kg TPG and 30 mg/kg DDC),with 18 rats in each,administered once a day for 7 consecutive days.After the administration,the neurological deficit score of the rats was evaluated.Nissl staining microscopy was applied to observe neuronal activity in brain tissue.2,3,5-triphenyltetrazolium chloride(TTC)staining microscopy was applied to detect the area of cerebral infarction in rats.The level of interleukin-1β and IL-18 in brain tissue was measured by ELISA method.Western blot was applied to detect the expression of purinergic receptor P2X ligand-gated ion channel 7(P2X7R)/NLRP3 signaling pathway related proteins and pyroptosis related proteins such as apoptosis associated speck like protein containing a CARD(ASC)and cysteine protease 1(caspase-1)proteins in brain tissue.Results Compared with the sham group,the neurological deficit score,infarct area,level of IL-1β and IL-18 in brain tissue and protein expression of P2X7R,NLRP3,ASC,and caspase-1 in brain tissue of rats in the simple CI/RI group were significantly increased(P<0.05),while the proportion of Nissl body positive cells in brain tissue was significantly reduced(P<0.05).The change in corresponding indicators of rats in the nimodipine group,TPG-L group,and TPG-H group was opposite to those in the simple CI/RI group(P<0.05).NLRP3 acti-vator DDC antagonized the inhibitory effect of TPG on cell pyroptosis in CI/RI rats.Conclusions TPG may inhibit brain injury in CI/RI rats by down-regulating the P2X7R/NLRP3 pathway.

Abstract Polycystic ovarian syndrome(PCOS) is a very common endocrine and reproductive disease. Its etiology and pathogenesis are complex and not yet fully clear. At present, the clinical treatment is mainly symptomatic. Studies have revealed that ferroptosis, as a new form of cell death, may play a key regulatory role in the occurrence and development of PCOS. In addition, there is an increase in autophagy/mitochondrial autophagy in PCOS patients, which may be closely related to the occurrence of ferroptosis. This review summarizes the pathogenesis of mitochondrial autophagy and ferroptosis in PCOS, and analyzes the interrelationship between mitochondrial autophagy and ferroptosis in granulosa cells, in order to provide new insights and potential therapeutic targets for the clinical treatment of PCOS.