ObjectiveCerebral ischemic injury triggers a complex pathological cascade characterized by excessive reactive oxygen species (ROS) accumulation, persistent oxidative stress, and sustained neuroinflammation in the injured brain microenvironment. These events collectively drive mitochondrial dysfunction, microglial overactivation, pro-inflammatory cytokine release, and progressive neuronal apoptosis, ultimately leading to severe and irreversible neurological deficits. However, conventional therapeutic strategies face critical limitations, including poor blood-brain barrier penetration, insufficient local drug concentration, uncontrolled drug release, and off-target systemic side effects. To address this pathological process, we rationally designed and fabricated an injectable ROS-responsive hydrogel loaded with polydopamine nanoparticles (PDA NPs) for spatiotemporally controlled antioxidation, anti-inflammation, and neuroprotection in the ischemic injury microenvironment. The present study aimed to systematically characterize the physicochemical properties, ROS-responsive drug release behavior, biocompatibility, and neuroprotective efficacy of this composite hydrogel system in vitro. MethodsPDA NPs were fabricated via oxidative self-polymerization. The ROS-responsive hydrogel was cross-linked using N1-(4-boronobenzyl)-N3-(4-boronophenyl)-N1,N1,N3,N3-tetramethylpropane-1, 3-diaminium (TSPBA) and polyvinyl alcohol (PVA). Morphology, particle size, Zeta potential, and structure of PDA NPs were characterized by dynamic light scattering (DLS), Zeta potential analysis, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Microstructure, rheological properties, shear-thinning behavior, and ROS-triggered release profiles of the hydrogel were examined by SEM and rheometry. Biocompatibility was evaluated using HT22 mouse hippocampal neurons with CCK-8 and live/dead staining. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was established to simulate ischemic injury in vitro. ROS levels and neuronal apoptosis were detected by DHE staining and TUNEL assay. Microglial polarization and pro-inflammatory cytokine expression were analyzed using immunofluorescence and RT-qPCR in BV-2 microglia. Transwell co-culture was used to verify the indirect neuroprotection mediated by modulated microglia. ResultsCharacterization results confirmed that the as-prepared PDA NPs were monodispersed spherical nanoparticles with uniform diameter and negative surface potential, demonstrating favorable dispersibility and robust ROS-scavenging activity. The TSPBA-PVA hydrogel exhibited a highly porous interconnected network, suitable mechanical strength, and obvious shear-thinning behavior, supporting its application as an injectable implant. More importantly, the hydrogel displayed typical ROS-responsive degradation and on-demand PDA NP release in a ROS-concentration-dependent manner. In vitro cellular experiments demonstrated that the PDA NP-loaded hydrogel possessed excellent biocompatibility with HT22 cells. In the OGD/R model, the hydrogel significantly reduced intracellular ROS accumulation and markedly suppressed neuronal apoptosis. Furthermore, the composite hydrogel effectively redirected BV-2 microglia from the pro-inflammatory M1 toward the anti-inflammatory M2 phenotypes, downregulated the expression of pro-inflammatory cytokines including TNF-α, IL-1β, and IL-6, and reduced inflammatory damage. Transwell co-culture assays further validated that M2-polarized microglia mediated by the hydrogel significantly enhanced the survival of OGD/R-injured HT22 neurons and attenuated apoptosis. ConclusionIn this study, we successfully developed a novel injectable ROS-responsive hydrogel loaded with PDA NPs for synergistic antioxidative and anti-inflammatory neuroprotection. This intelligent hydrogel system enables ROS-triggered on-demand release of PDA NPs, efficiently scavenges excessive ROS, inhibits oxidative stress injury, modulates microglial polarization, and suppresses neuroinflammation, thereby exerting robust neuroprotective effects in vitro. This biomaterial platform provides a promising strategy for the targeted and controlled delivery of bioactive nanomaterials in the central nervous system diseases and establishes a solid experimental foundation for the development of in situ injectable therapies for ischemic brain injury.

Objective:To analyze the changes and significance in clinical cardiac indicators of early cardiac myocardial dysfunction and cardiac substructure dose during conventional radiotherapy for N 2-N 3 non-small cell lung cancer (NSCLC) with mediastinal lymph node metastases. Methods:The data of 34 NSCLC patients with lymph node metastases in regions 4-8 admitted to the Affiliated Cancer Hospital of Guizhou Medical University from June 2022 to August 2023 were observed and analyzed. All patients were treated with volumetric modulated arc therapy with a prescribed dose of 60-70 Gy. Cardiac troponin T (cTnT) and N-terminal pro-B-type natriuretic peptide (NT-proBNP) were measured at 6 time points: within 1 week before radiotherapy ( t0); when the cumulative radiotherapy dose reaches 20 Gy ( t20), 40 Gy ( t40), 60 Gy ( t60) during radiotherapy; within 1 week after radiotherapy ( tp); 1 month after radiotherapy( tp1). Left ventricular global longitudinal strain (LVGLS) and left atrial global longitudinal strain (LAGLS) were assessed at 4 time points: t0, t40, tp and tp1, respectively. The changes in cardiac indicators at different time points during radiotherapy and their correlation with substructure doses were analyzed using analysis of variance, linear regression analysis, and Pearson correlation. Results:The correlation between cardiac substructure dose and mean heart dose (MHD) in the study cohort in the descending order was as follows: left ventricle ( B=0.43, P<0.001), right ventricle ( B=0.37, P=0.002), left atrium ( B=0.16, P<0.001), and right atrium ( B=0.15, P=0.001). There were significant differences in the changes of LVGLS and LAGLS across different time points ( F=3.13, P=0.029; F=17.18, P<0.001). At 1 month after radiation, LAGLS was significantly decreased compared to pre-radiation levels ( P=0.009), whereas no significant difference was observed in LVGLS ( P=1.000). No significant differences were observed in the changes of cTnT and NT-proBNP across different time points (all P>0.05). Significant correlations were identified between cTnT and right ventricle mean dose at t40 ( r=0.38, P=0.025), as well as between NT-proBNP and right atrium mean dose at t60 and tp ( r=0.54, P=0.001; r=0.41, P=0.016). Conclusions:At present, there is no significant difference between the sensitive serum markers of myocardial injury and LVGLS in detecting early myocardial injury. LAGLS may hold substantial clinical value. There is uncertainty about radiation injury and repair of various cardiac substructures.

Objective:To analyze the changes and significance in clinical cardiac indicators of early cardiac myocardial dysfunction and cardiac substructure dose during conventional radiotherapy for N 2-N 3 non-small cell lung cancer (NSCLC) with mediastinal lymph node metastases. Methods:The data of 34 NSCLC patients with lymph node metastases in regions 4-8 admitted to the Affiliated Cancer Hospital of Guizhou Medical University from June 2022 to August 2023 were observed and analyzed. All patients were treated with volumetric modulated arc therapy with a prescribed dose of 60-70 Gy. Cardiac troponin T (cTnT) and N-terminal pro-B-type natriuretic peptide (NT-proBNP) were measured at 6 time points: within 1 week before radiotherapy ( t0); when the cumulative radiotherapy dose reaches 20 Gy ( t20), 40 Gy ( t40), 60 Gy ( t60) during radiotherapy; within 1 week after radiotherapy ( tp); 1 month after radiotherapy( tp1). Left ventricular global longitudinal strain (LVGLS) and left atrial global longitudinal strain (LAGLS) were assessed at 4 time points: t0, t40, tp and tp1, respectively. The changes in cardiac indicators at different time points during radiotherapy and their correlation with substructure doses were analyzed using analysis of variance, linear regression analysis, and Pearson correlation. Results:The correlation between cardiac substructure dose and mean heart dose (MHD) in the study cohort in the descending order was as follows: left ventricle ( B=0.43, P<0.001), right ventricle ( B=0.37, P=0.002), left atrium ( B=0.16, P<0.001), and right atrium ( B=0.15, P=0.001). There were significant differences in the changes of LVGLS and LAGLS across different time points ( F=3.13, P=0.029; F=17.18, P<0.001). At 1 month after radiation, LAGLS was significantly decreased compared to pre-radiation levels ( P=0.009), whereas no significant difference was observed in LVGLS ( P=1.000). No significant differences were observed in the changes of cTnT and NT-proBNP across different time points (all P>0.05). Significant correlations were identified between cTnT and right ventricle mean dose at t40 ( r=0.38, P=0.025), as well as between NT-proBNP and right atrium mean dose at t60 and tp ( r=0.54, P=0.001; r=0.41, P=0.016). Conclusions:At present, there is no significant difference between the sensitive serum markers of myocardial injury and LVGLS in detecting early myocardial injury. LAGLS may hold substantial clinical value. There is uncertainty about radiation injury and repair of various cardiac substructures.

Objective:To investigate the expression and clinical significance of gamma-aminobutyric acid receptor-associated protein-like 1 (GABARAPL1) in patients with gouty arthritis (GA).Methods:Differentially expressed genes (DEGs) between GA patients and healthy controls were identified by the Gene Expression Omnibus (GEO) database, followed by the screening of autophagy-related DEGs (au-DEGs) associated with GA. Peripheral blood single nucleated cells (PBMCs) were collected from 80 male GA patients and 60 healthy controls treated at the Affiliated Taizhou People′s Hospital of Nanjing Medical University between March to December 2023. Real-time quantitative PCR (qRT-PCR) was employed to assess the expression of au-DEGs, bisulfite sequencing PCR (BSP) was used to determine the methylation status of the GABARAPL1 promoter, and chromatin immunoprecipitation combined with qPCR (ChIP-qPCR) was used to analyze the binding activity of histone H3 to the GABARAPL1 promoter. Spearman ′s Correlations between GABARAPL1 expression and clinical indicators such as serum uric acid (UA) were analyzed. The diagnostic efficacy of GABARAPL1 in PBMCs for GA was evaluated using receiver operating characteristic (ROC) curves. Results:The relative expression of GABARAPL1 in PBMCs was significantly higher in GA patients compared to healthy controls (1.91±0.72 vs. 0.84±0.39, t=11.27, P<0.001). Spearman correlation analysis revealed that GABARAPL1 expression was positively correlated with UA, white blood cell count, neutrophil count, monocyte count, globulin, urea, and creatinine ( r=0.61, 0.40, 0.46, 0.32, 0.28, 0.22, 0.34; P<0.05 for all), and negatively correlated with lymphocyte count, albumin, high-density lipoprotein cholesterol, apolipoprotein A1, and estimated glomerular filtration rate ( r=-0.18, -0.34, -0.33, -0.50, -0.22; P<0.05 for all). ROC curve analysis showed that GABARAPL1 in PBMCs had high diagnostic efficacy for GA, with an area under the curve of 0.936. No significant differences in the methylation levels of GABARAPL1 promoter regions MC1 and MC2 were observed between GA patients and healthy controls ( t=2.28、1.43, P<0.05 for all), whereas histone H3 acetylation levels were significantly elevated in GA patients ( t=11.19, P<0.001). Conclusion:The expression of GABARAPL1 in PBMCs is significantly upregulated in GA patients, which may be associated with increased histone H3 acetylation at its promoter. The high expression of GABARAPL1 may contribute to the pathogenesis of GA by regulating autophagy and holds promise as a diagnostic biomarker and potential therapeutic target for GA.

Objective To compare the value of standard deviation(SD)method and root mean square(RMS)method for measuring signal-to-noise ratio(SNR)of MRI of American College of Radiology(ACR)phantom based on phased array coils.Methods ACR phantom was scanned with head coil(Head),abdominal coil(Anterior),dual piece flexible coil(Flex L)and abdominal coil+flexible coil(Anterior+Flex L)each for 6 times,respectively,with sequences of axial T 1W-spin echo(SE)and T2W-SE.And the scanning schemes were divided into 8 groups based on 4 types of coils and 2 sequences.SNR of 8 groups were measured with standard deviation method and root mean square method,respectively,and the differences between the above 2 methods were observed.Results For T1W-SE sequence,SNRsD of MRI based on Head,Anterior,Flex L and Anterior+Flex L was 1.25±0.07,0.56±0.02,1.15±0.10 and 1.04±0.11,respectively,while SNRRMs was 1.10±0.07,0.58±0.03,1.01±0.13 and 1.31±0.15,respectively.For T2W-SE sequence,SNRsD was 1.40±0.08,0.48±0.02,1.04±0.07 and 1.08±0.06,respectively,while SNRRMs was 1.29±0.09,0.53±0.03,0.84±0.08 and 1.35±0.12,respectively.Compared pairwise,significant difference of SNRsD was found in 9 pairs(all P＜0.05),while of SNRRMs was detected in 10 pairs(all P＜0.05).Conclusion Compared with standard deviation method,root mean square method was more suitable for measuring SNR of MRI of ACR phantom based on phased array coils.

Objective To establish a flow cytometry method for detecting C1s protein on platelet surface and preliminarily explore its potential application value in the auxiliary diagnosis of primary immune thrombocytopenia(ITP).Methods C1s-conjugated 2 μm car-boxylated magnetic beads(C1s beads)were prepared and used as quality control particles.Fluorescein isothiocyanate(FITC)-labeled anti-C1s antibody was employed as the detection antibody to develop a flow cytometric assay for detecting C1s protein expression on platelets.The intra-assay and inter-assay precision,as well as the dilution linearity of the method,were evaluated.Subsequently,the expression levels of C1 s protein on the surface of platelets were compared among the ITP group,the non-ITP thrombocytopenia group,and the healthy control group.Results Light microscopy showed that both unconjugated carboxylated magnetic beads(blank beads)and C1s-conjugated beads were uniformly dispersed without aggregation.Under fluorescence microscopy,C1s beads exhibited strong yellow-green fluorescence,whereas the blank beads showed no fluorescence signal.The established flow cytometry assay exhibited ac-ceptable precision,with intra-assay coefficient of variation(CV)values of 7.02％,7.12％,and 3.91％for low,medium,and high con-centrations of C1s beads,respectively,and inter-assay CV values of 13.49％,6.15％,and 0.78％,respectively.The dilution linearity was satisfactory,coefficient of determination(R2)=0.998 8.Clinical sample testing revealed that the proportion of C1s-positive plate-lets in ITP group(2.56±0.79)％was significantly higher than that in healthy control group(0.23±0.18)％and the non-ITP thrombo-cytopenia control group(0.22±0.10)％,with statistically significant differences(both P＜0.05).Conclusion This study successfully established a stable and reliable flow-cytometry method for quantifying C1s expression on platelet surface and preliminarily demonstrated that C1s expression is significantly elevated on platelets of ITP patients,suggesting that C1s could serve as a potential auxiliary diag-nostic marker for ITP.

Objective To detect the content of 5'-tRF-GlyGCC in peripheral blood and explore its applica-tion value in the early diagnosis of colorectal cancer(CRC).Methods Peripheral blood samples were randomly collected from 99 CRC patients,11 patients with enteritis,17 patients with colorectal polyps and 99 healthy indi-viduals in the same period at the Affiliated Hospital of Guizhou Medical University.The content of 5'-tRF-GlyGCC in total RNA of peripheral blood was detected by fluorescence RT-PCR.The receiver operating characteristic curve(ROC)was established to analyze the value of 5'-tRF-GlyGCC content in the early diagnosis of CRC and to explore the relationship between its content and the clinical and pathological characteristics of CRC patients.Results The content of 5'-tRF-GlyGCC in the peripheral blood of CRC patients and colorectal polyp patients was significantly higher than that of healthy individuals and patients with enteritis,and the differences were statistically significant(P<0.05).The area under the curve(AUC)of 5'-tRF-GlyGCC for diagnosing CRC was 0.817(95%CI:0.763～0.871),with a diagnostic sensitivity of 56.7%and a specificity of 99.9%.The content of 5'-tRF-GlyGCC in peripheral blood was significantly positively correlated with the levels of CEA and CA19-9 in CRC patients.Conclusion The detection of 5'-tRF-GlyGCC in peripheral blood has a high application value in the early diagnosis of CRC and can be used for clinical reference and research.

Objective To explore the diagnostic value of exhaled volatile organic compounds (VOCs) for cystic fibrosis (CF). Methods A systematic search was conducted in PubMed, EMbase, Web of Science, Cochrane Library, CNKI, Wanfang, VIP, and SinoMed databases up to August 7, 2024. Studies that met the inclusion criteria were selected for data extraction and quality assessment. The quality of included studies was assessed by the Newcastle-Ottawa Scale (NOS), and the risk of bias and applicability of included prediction model studies were assessed by the prediction model risk of bias assessment tool (PROBAST). Results A total of 10 studies were included, among which 5 studies only identified specific exhaled VOCs in CF patients, and another 5 developed 7 CF risk prediction models based on the identification of VOCs in CF. The included studies reported a total of 75 exhaled VOCs, most of which belonged to the categories of acylcarnitines, aldehydes, acids, and esters. Most models (n=6, 85.7%) only included exhaled VOCs as predictive factors, and only one model included factors other than VOCs, including forced expiratory flow at 75% of forced vital capacity (FEF75) and modified Medical Research Council scale for the assessment of dyspnea (mMRC). The accuracy of the models ranged from 77% to 100%, and the area under the receiver operating characteristic curve ranged from 0.771 to 0.988. None of the included studies provided information on the calibration of the models. The results of the Prediction Model Risk of Bias Assessment Tool (PROBAST) showed that the overall bias risk of all predictive model studies was high, and the overall applicability was unclear. Conclusion The exhaled VOCs reported in the included studies showed significant heterogeneity, and more research is needed to explore specific compounds for CF. In addition, risk prediction models based on exhaled VOCs have certain value in the diagnosis of CF, but the overall bias risk is relatively high and needs further optimization from aspects such as model construction and validation.

Objective To compare the value of standard deviation(SD)method and root mean square(RMS)method for measuring signal-to-noise ratio(SNR)of MRI of American College of Radiology(ACR)phantom based on phased array coils.Methods ACR phantom was scanned with head coil(Head),abdominal coil(Anterior),dual piece flexible coil(Flex L)and abdominal coil+flexible coil(Anterior+Flex L)each for 6 times,respectively,with sequences of axial T 1W-spin echo(SE)and T2W-SE.And the scanning schemes were divided into 8 groups based on 4 types of coils and 2 sequences.SNR of 8 groups were measured with standard deviation method and root mean square method,respectively,and the differences between the above 2 methods were observed.Results For T1W-SE sequence,SNRsD of MRI based on Head,Anterior,Flex L and Anterior+Flex L was 1.25±0.07,0.56±0.02,1.15±0.10 and 1.04±0.11,respectively,while SNRRMs was 1.10±0.07,0.58±0.03,1.01±0.13 and 1.31±0.15,respectively.For T2W-SE sequence,SNRsD was 1.40±0.08,0.48±0.02,1.04±0.07 and 1.08±0.06,respectively,while SNRRMs was 1.29±0.09,0.53±0.03,0.84±0.08 and 1.35±0.12,respectively.Compared pairwise,significant difference of SNRsD was found in 9 pairs(all P＜0.05),while of SNRRMs was detected in 10 pairs(all P＜0.05).Conclusion Compared with standard deviation method,root mean square method was more suitable for measuring SNR of MRI of ACR phantom based on phased array coils.

Testicular histology based on testicular biopsy is an important factor for determining appropriate testicular sperm extraction surgery and predicting sperm retrieval outcomes in patients with azoospermia. Therefore, we developed a deep learning (DL) model to establish the associations between testicular grayscale ultrasound images and testicular histology. We retrospectively included two-dimensional testicular grayscale ultrasound from patients with azoospermia (353 men with 4357 images between July 2017 and December 2021 in The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China) to develop a DL model. We obtained testicular histology during conventional testicular sperm extraction. Our DL model was trained based on ultrasound images or fusion data (ultrasound images fused with the corresponding testicular volume) to distinguish spermatozoa presence in pathology (SPP) and spermatozoa absence in pathology (SAP) and to classify maturation arrest (MA) and Sertoli cell-only syndrome (SCOS) in patients with SAP. Areas under the receiver operating characteristic curve (AUCs), accuracy, sensitivity, and specificity were used to analyze model performance. DL based on images achieved an AUC of 0.922 (95% confidence interval [CI]: 0.908-0.935), a sensitivity of 80.9%, a specificity of 84.6%, and an accuracy of 83.5% in predicting SPP (including normal spermatogenesis and hypospermatogenesis) and SAP (including MA and SCOS). In the identification of SCOS and MA, DL on fusion data yielded better diagnostic performance with an AUC of 0.979 (95% CI: 0.969-0.989), a sensitivity of 89.7%, a specificity of 97.1%, and an accuracy of 92.1%. Our study provides a noninvasive method to predict testicular histology for patients with azoospermia, which would avoid unnecessary testicular biopsy.
Humans ; Male ; Azoospermia/diagnostic imaging* ; Deep Learning ; Testis/pathology* ; Retrospective Studies ; Adult ; Ultrasonography/methods* ; Sperm Retrieval ; Sertoli Cell-Only Syndrome/diagnostic imaging*