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/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.

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.

Objective:To retrieve, evaluate and integrate the evidence related to postoperative exercise rehabilitation for hip fracture in elderly frail patients, and provide reference for clinical rehabilitation nursing.Methods:Evidence on postoperative exercise rehabilitation of hip fractures in elderly frail patients was systemically retrieved in the guideline websites, professional association websites and databases, such as BMJ Best Practice, UpToDate, PubMed, Web of Science, CINAHL, Cochrane Library, Embase, Chinese Biomedical Database, China National Knowledge Infrastructure, Wanfang and VIP, including Best practices, guidelines, clinical decisions, evidence summaries, systematic reviews, and expert consensus. The retrieval time was from the establishment of the database to March 31, 2024.Results:A total of 33 articles were involved, including 12 guidelines, 2 clinical decisions, 7 systematic reviews, 7 expert consensuses, 5 evidence summaries. Forty pieces of evidence were summarized in 7 aspects: the role of exercise, exercise assessment, exercise planning, exercise content, exercise intensity, exercise adherence, exercise nursing, exercise guidance.Conclusions:Healthcare professionals should combine patients' tolerance, rehabilitation goals, and specific clinical situations to target and guide patients in exercise rehabilitation to promote the recovery of postoperative limb function, mitigate frailty, and improve the quality of life.

The clinical antiprotozoal drug nitazoxanide has been demonstrated to improve the experimental diabetes mellitus, lipid metabolism disorders, atherosclerosis and inhibit inflammation. Since the pathogenesis of heart failure with preserved ejection (HFpEF) is multifactorial and closely associated with the aforementioned diseases, we aim to study the effect of nitazoxanide on high-fat diet (HFD) plus L-NAME (N ω-nitro-l-arginine methyl ester)-induced HFpEF and metabolic syndrome in mice. We found that oral nitazoxanide improved cardiac hypertrophy, cardiac fibrosis, cardiac diastolic dysfunction, increased blood pressure, impaired exercise tolerance, impaired glucose handling, serum lipid disorders, hepatic steatosis, increased weight of white adipose tissues and kidney fibrosis in HFD + L-NAME-treated mice. In the established HFD + L-NAME-induced HFpEF and metabolic syndrome mouse model, therapeutic treatment with nitazoxanide rescued HFD + L-NAME-induced pathological phenotypes as mentioned above. The in vitro experiments revealed that tizoxanide, the active metabolite of nitazoxanide, increased the basal mitochondria metabolism of cardiomyocytes, inhibited cardiomyocyte hypertrophy and collagen secretion from cardiac fibroblasts, and relaxed phenylephrine- and U46619-induced constriction of rat mesenteric arteries, indicating that the direct effect of tizoxanide might partly contribute to the protective effect of nitazoxanide against HFpEF in vivo. The present study suggests that nitazoxanide might be a potential drug for HFpEF and metabolic syndrome therapy.

OBJECTIVES: To investigate the impact of prenatal fear stress on placental amino acid transport and emotion and cognition development in offspring rats. METHODS: Thirty pregnant Wistar rats were randomized equally into control and fear stress (induced using an observational foot shock model) groups. In each group, placental and serum samples were collected from 6 dams on gestational day 20, and the remaining rats delivered naturally and the offspring rats were raised under the same conditions until 8 weeks of age. Emotional and cognitive outcomes of the offspring rats were assessed with behavioral tests, and placental structure was examined using HE staining. Bioinformatics analysis was used to identify differentially expressed placental transporter genes under fear stress. The expressions of system A and system L amino acid transporters, along with other specialized transporters, were detected using qRT-PCR and Western blotting. Fetal serum amino acid concentrations were determined by HPLC. The correlations between fetal amino acid levels and behavioral outcomes of the offspring rats were analyzed. RESULTS: The dams with fear stress showed reduced open-field activity and increased freezing behavior with significantly decreased placental weight, fetal weight, and fetal-to-placental ratio. Bioinformatics analysis revealed 28 differentially expressed transporter genes involved mainly in amino acid transport. In the fear stress group, fetal serum amino acid levels were significantly lowered and Slc38a1, Slc43a1, Slc43a2, Slc7a8, Slc6a6, Slc1a1 and Slc6a9 mRNA and protein expressions were all downregulated. The offspring rats in fear stress group exhibited decreased novel object preference and spontaneous alternation with reduced open arm exploration and increased immobility in emotional tests. Lower early-life amino acid levels was found to correlate with impaired adult cognition. CONCLUSIONS Prenatal fear stress in rats impairs placental amino acid transporter expression and reduces fetal serum amino acid levels, potentially contributing to long-term cognitive deficits in the offspring rats.
Animals ; Female ; Pregnancy ; Placenta/metabolism* ; Fear ; Rats ; Rats, Wistar ; Cognition ; Prenatal Exposure Delayed Effects ; Stress, Psychological ; Amino Acids/blood* ; Amino Acid Transport Systems/metabolism*

Depression is increasingly prevalent among adolescents and can profoundly impact their lives. However, the early detection of depression is often hindered by the time-consuming diagnostic process and the absence of objective biomarkers. In this study, we propose a novel approach for depression detection based on an affective brain-computer interface (aBCI) and the resting-state electroencephalogram (EEG). By fusing EEG features associated with both emotional and resting states, our method captures comprehensive depression-related information. The final depression detection model, derived through decision fusion with multiple independent models, further enhances detection efficacy. Our experiments involved 40 adolescents with depression and 40 matched controls. The proposed model achieved an accuracy of 86.54% on cross-validation and 88.20% on the independent test set, demonstrating the efficiency of multimodal fusion. In addition, further analysis revealed distinct brain activity patterns between the two groups across different modalities. These findings hold promise for new directions in depression detection and intervention.
Humans ; Male ; Female ; Adolescent ; Case-Control Studies ; Depression/diagnosis* ; Early Diagnosis ; Rest ; Electroencephalography/methods* ; Brain-Computer Interfaces ; Models, Psychological ; Reproducibility of Results ; Affect/physiology* ; Photic Stimulation/methods* ; Video Recording ; Brain/physiopathology*

Rapid and ultrasensitive detection of pathogen-associated biomarkers is vital for the early diagnosis and therapy of bacterial infections. Herein, we developed a close-packed and ordered Au@AgPt array coupled with a cascade triggering strategy for surface-enhanced Raman scattering (SERS) and colorimetric identification of the Staphylococcus aureus biomarker micrococcal nuclease (MNase) in serum samples. The trimetallic Au@AgPt nanozymes can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) molecules to SERS-enhanced oxidized TMB (oxTMB), accompanied by the color change from colorless to blue. In the presence of S. aureus, the secreted MNase preferentially cut the nucleobase AT-rich regions of DNA sequences on magnetic beads (MBs) to release alkaline phosphatase (ALP), which subsequently mediated the oxTMB reduction for inducing the colorimetric/SERS signal fade away. Using this "on-to-off" triggering strategy, the target S. aureus can be recorded in a wide linear range with a limit of detection of 38 CFU/mL in the colorimetric mode and 6 CFU/mL in the SERS mode. Meanwhile, the MNase-mediated strategy characterized by high specificity and sensitivity successfully discriminated between patients with sepsis (n = 7) and healthy participants (n = 3), as well as monitored the prognostic progression of the disease (n = 2). Overall, benefiting from highly active and dense "hot spot" substrate, MNase-mediated cascade response strategy, and colorimetric/SERS dual-signal output, this methodology will offer a promising avenue for the early diagnosis of S. aureus infection.

Objective:To investigate the rehabilitation effect of Xingyi Baduanjin training on cardiac function and quality of life in postoperative percutaneous coronary intervention(PCI)patients with coronary heart disease.Method:From September 2017 to January 2021,a total of 119 postoperative PCI patients with coronary heart disease in the Department of Cardiovascular Medicine of the First Medical Center of the General Hospi-tal of the People's Liberation Army(PLA)were selected and randomly divided into the experimental group(n=59)and the control group(n=60).In the control group,a conventional cardiac rehabilitation program was executed,while in the experimental group,a rehabilitation program combining the conventional program with Xingyi Baduanjin training was executed.The treatment time was 6 weeks.Before and after the 6 weeks,the exercise endurance of the two groups was measured by cardiopulmonary exercise test,which mainly observed the changes in peak power,peak oxygen consumption,peak metabolic equivalent,peak oxygen pulse,and peak respiratory exchange rhythm.The Quality of Life Scale Brief Version(QOL-BREF)and the MOS 36-Item Short Form Health Survey(SF-36)were used to assess the quality of life and health status in patients.Result:Peak power,peak oxygen consumption,peak metabolic equivalents,peak oxygen pulse and peak re-spiratory exchange rate were not significantly different between the two groups before treatment(P>0.05).Peak power,peak oxygen consumption,and peak metabolic equivalents in both groups after 6 weeks of treat-ment were significantly higher than before treatment(t>3.141,P<0.05),and the experimental group was signif-icantly higher than the control group(t>2.017,P<0.05).Peak oxygen pulse in the experimental group was sig-nificantly higher than before treatment(t=7.459,P<0.05)and was significantly higher than that in the control group(t=5.866,P<0.05).The respiratory exchange rate was significantly lower than before treatment(t=6.281,P<0.05)and was significantly lower than that in the control group(t=2.686,P<0.05).The QOL-BREF scores were not significantly different between the two groups before treatment(P>0.05).The QOL-BREF scores in both groups after 6 weeks of treatment were significantly higher than before treatment(t>23.620,P<0.05),and the experimental group was significantly higher than the control group(t=12.719,P<0.05).The SF-36 scores were not significantly different between the two groups before treatment(P>0.05).The SF-36 scores in both groups after 6 weeks of treatment were significantly higher than before treatment(t>35.255,P<0.05),and the experimental group was significantly higher than the control group(t=16.075,P<0.05).Conclusion:Xingyi Baduanjin training can effectively enhance exercise endurance,which improves the quality of life and health status in postoperative PCI patients with coronary artery disease,and achieves a good reha-bilitation effect.

Objective To investigate the effect of long non-coding RNA small nucleolar RNA host gene 14(LncRNA SNHG14)on high glucose-induced podocyte injury by targeting microRNA-30a-5p(miR-30a-5p).Methods Podocytes were cultured in vitro and were divided into the following groups:the standard glucose(NG)group,the high glucose(HG)group,the si-NC+HG group,the si-SNHG14+HG group,the miR-NC+HG group,the miR-30a-5p mimics+HG group,the si-SNHG14+inhibitor NC+HG group and the si-SNHG14+miR-30a-5p inhibitor+HG group.Quantitative real-time polymerase chain reaction(RT-qPCR)was performed to detect expression levels of LncRNA SNHG14 and miR-30a-5p.Flow cytometry was used to determine cell apoptosis.Enzyme-linked immunosorbent assay(ELISA)was applied to measure levels of tumor necrosis factor-alpha(TNF-α),interleukin(IL)-6 and IL-1β.Xanthine oxidase method,ammonium molybdate colorimetry and thiobarbituric acid method were respectively used to detect superoxide dismutase(SOD),catalase(CAT)and malondialdehyde(MDA).Dual-luciferase reporter gene was conducted to verify the targeting relationship between LncRNA SNHG14 and miR-30a-5p.Western blot assay was performed to detect expression levels of apoptosis-related proteins.Results Compared with the NG group,the HG group exhibited increased expression levels of LncRNA SNHG14,cell apoptosis rate,as well as levels of TNF-α,IL-6,IL-1β and MDA,whereas the expression level of miR-30a-5p and levels of SOD and CAT were decreased(P＜0.05).Compared with the HG group and the si-NC+HG group,the si-SNHG14+HG group exhibited decreased expression levels of LncRNA SNHG14,apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins,while the expression level of miR-30a-5p,levels of SOD and CAT and the expression level of Bcl-2 protein were increased(P＜0.05).Compared with the HG group and the miR-NC+HG group,the miR-30a-5p mimics+HG group showed no significant difference in the expression level of LncRNA SNHG14(P＞0.05).Meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were increased,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were decreased(P＜0.05).Compared with the si-SNHG14+HG group and the si-SNHG14+inhibitor NC+HG group,the si-SNHG14+miR-30a-5p inhibitor+HG group showed no significant difference in the expression level of LncRNA SNHG14(P＞0.05),meanwhile,the expression level of miR-30a-5p,levels of SOD and CAT,and expression level of Bcl-2 protein were reduced,whereas the cell apoptosis rate,levels of TNF-α,IL-6,IL-1β and MDA,as well as expression levels of Bax and cleaved caspase-3 proteins were increased(P＜0.05).The dual-luciferase reporter gene assay confirmed that LncRNA SNHG14 targeted and negatively regulated miR-30a-5p.Conclusion The inhibition of LncRNA SNHG14 can target miR-30a-5p to alleviate high glucose induced podocyte injury.