MSC-derived small extracellular vesicles mitigate diabetic retinopathy by stabilizing Nrf2 through miR-143-3p-mediated inhibition of neddylation.

Diabetic retinopathy (DR) is a highly hazardous and widespread complication of diabetes mellitus (DM). The accumulated reactive oxygen species (ROS) play a central role in DR development. The aim of this research was to examine the impact and mechanisms of mesenchymal stem cell (MSC)-derived small extracellular vesicles (sEV) on regulating ROS and retinal damage in DR. Intravitreal injection of sEV inhibited Cullin3 neddylation, stabilized Nrf2, decreased ROS, reduced retinal inflammation, suppressed Müller gliosis, and mitigated DR. Based on MSC-sEV miRNA sequencing, bioinformatics software, and dual-luciferase reporter assay, miR-143-3p was identified to be the key effector for MSC-sEV's role in regulating neural precursor cell expressed developmentally down-regulated 8 (NEDD8)-mediated neddylation. sEV were able to be internalized by Müller cells. Compared to advanced glycation end-products (AGEs)-induced Müller cells, sEV coculture decreased Cullin3 neddylation, activated Nrf2 signal pathway to combat ROS-induced inflammation. The barrier function of endothelial cells was impaired when endothelial cells were treated with the supernatant of AGEs-induced Müller cells, but was restored when treated with supernatant of AGEs-induced Müller cells cocultured with sEV. The protective effect of sEV was, however, compromised when miR-143-3p was inhibited in sEV. Moreover, the protective efficacy of sEV was diminished when NEDD8 was overexpressed in Müller cells. These findings showed MSC-sEV delivered miR-143-3p to inhibit Cullin3 neddylation, stabilizing Nrf2 to counteract ROS-induced inflammation and reducing vascular leakage. Our findings suggest that MSC-sEV may be a potential nanotherapeutic agent for DR, and that Cullin3 neddylation could be a new target for DR therapy.

IVIM and DCE-MRI in Predicting Phenotypic Subtypes and Nottingham Prognostic Index of Breast Cancer.

OBJECTIVE: To explore the value of intravoxel incoherent motion (IVIM) and dynamic contrast enhanced MRI (DCE-MRI) for predicting phenotypic subtypes and Nottingham prognostic index (NPI) of breast cancer. STUDY DESIGN: Descriptive study. Place and Duration of the Study: Department of Radiology, Affiliated Hospital of Jining Medical University, Jining, Shandong, China, from March 2020 to January 2022. METHODOLOGY: One hundred and forty-one breast cancer patients with preoperative IVIM and DCE imaging were collected. IVIM parameters of D, D*, f, and DCE-MRI parameters of Ktrans, Kep, and Ve were measured. Receiver operating characteristic curves were conducted to assess the diagnostic efficacies. Additionally, 40 patients collected from February 2022 to July 2022 were enrolled as validation cohort. RESULTS: The D value in HER2-enriched (HER2-E) was lower than that in non-HER-E, while D*, Ktrans, and Ve values were higher than that in non-HER-E (p < 0.001, 0.046, < 0.001, and < 0.001, respectively). D + Ktrans + Ve showed an optimal diagnostic efficiency (AUC = 0.868). Meanwhile, D* and f values of triple-negative breast cancer (TNBC) were higher than those of non-TNBC, and Ve value of TNBC was lower than that of non-TNBC (p = 0.013, 0.006, and < 0.001, respectively). D* + f + Ve showed the best prediction performance (AUC = 0.849). Additionally, D and Kep were independent predictors of NPI (p < 0.001, and 0.002, respectively). D + Kep showed a good diagnostic efficiency (AUC = 0.818). CONCLUSION: The combined IVIM and DCE-MRI model showed enhanced diagnostic efficiency in predicting phenotypic subtypes and NPI of breast cancer, and might thus be considered efficient in therapy decision-making for patients. KEY WORDS: Breast neoplasms, Intravoxel incoherent motion, Dynamic contrast enhanced magnetic resonance imaging, Phenotypic subtypes, Nottingham prognostic index.

Preoperative differentiation of gastric schwannomas and gastrointestinal stromal tumors based on computed tomography: a retrospective multicenter observational study.

Introduction: Gastric schwannoma is a rare benign tumor accounting for only 1-2% of alimentary tract mesenchymal tumors. Owing to their low incidence rate, most cases are misdiagnosed as gastrointestinal stromal tumors (GISTs), especially tumors with a diameter of less than 5 cm. Therefore, this study aimed to develop and validate a diagnostic nomogram based on computed tomography (CT) imaging features for the preoperative prediction of gastric schwannomas and GISTs (diameters = 2-5 cm). Methods: Gastric schwannomas in 47 patients and GISTs in 230 patients were confirmed by surgical pathology. Thirty-four patients with gastric schwannomas and 167 with GISTs admitted between June 2009 and August 2022 at Hospital 1 were retrospectively analyzed as the test and training sets, respectively. Seventy-six patients (13 with gastric schwannomas and 63 with GISTs) were included in the external validation set (June 2017 to September 2022 at Hospital 2). The independent factors for differentiating gastric schwannomas from GISTs were obtained by multivariate logistic regression analysis, and a corresponding nomogram model was established. The accuracy of the nomogram was evaluated using receiver operating characteristic and calibration curves. Results: Logistic regression analysis showed that the growth pattern (odds ratio [OR] 3.626; 95% confidence interval [CI] 1.105-11.900), absence of necrosis (OR 4.752; 95% CI 1.464-15.424), presence of tumor-associated lymph nodes (OR 23.978; 95% CI 6.499-88.466), the difference between CT values during the portal and arterial phases (OR 1.117; 95% CI 1.042-1.198), and the difference between CT values during the delayed and portal phases (OR 1.159; 95% CI 1.080-1.245) were independent factors in differentiating gastric schwannoma from GIST. The resulting individualized prediction nomogram showed good discrimination in the training (area under the curve [AUC], 0.937; 95% CI, 0.900-0.973) and validation (AUC, 0.921; 95% CI, 0.830-1.000) datasets. The calibration curve showed that the probability of gastric schwannomas predicted using the nomogram agreed well with the actual value. Conclusion: The proposed nomogram model based on CT imaging features can be used to differentiate gastric schwannoma from GIST before surgery.

Blockade of the lncRNA-DOT1L-LAMP5 axis enhances autophagy and promotes degradation of MLL fusion proteins.

BACKGROUND: Mixed-lineage leukemia (MLL) fusion gene caused by chromosomal rearrangement is a dominant oncogenic driver in leukemia. Due to having diverse MLL rearrangements and complex characteristics, MLL leukemia treated by currently available strategies is frequently associated with a poor outcome. Therefore, there is an urgent need to identify novel therapeutic targets for hematological malignancies with MLL rearrangements. METHODS: qRT-PCR, western blot, and spearman correction analysis were used to validate the regulation of LAMP5-AS1 on LAMP5 expression. In vitro and in vivo experiments were conducted to assess the functional relevance of LAMP5-AS1 in MLL leukemia cell survival. We utilized chromatin isolation by RNA purification (ChIRP) assay, RNA pull-down assay, chromatin immunoprecipitation (ChIP), RNA fluorescence in situ hybridization (FISH), and immunofluorescence to elucidate the relationship among LAMP5-AS1, DOT1L, and the LAMP5 locus. Autophagy regulation by LAMP5-AS1 was evaluated through LC3B puncta, autolysosome observation via transmission electron microscopy (TEM), and mRFP-GFP-LC3 puncta in autophagic flux. RESULTS: The study shows the crucial role of LAMP5-AS1 in promoting MLL leukemia cell survival. LAMP5-AS1 acts as a novel autophagic suppressor, safeguarding MLL fusion proteins from autophagic degradation. Knocking down LAMP5-AS1 significantly induced apoptosis in MLL leukemia cell lines and primary cells and extended the survival of mice in vivo. Mechanistically, LAMP5-AS1 recruits the H3K79 histone methyltransferase DOT1L to LAMP5 locus, directly activating LAMP5 expression. Importantly, blockade of LAMP5-AS1-LAMP5 axis can represses MLL fusion proteins by enhancing their degradation. CONCLUSIONS: The findings underscore the significance of LAMP5-AS1 in MLL leukemia progression through the regulation of the autophagy pathway. Additionally, this study unveils the novel lncRNA-DOT1L-LAMP5 axis as promising therapeutic targets for degrading MLL fusion proteins.

MSC-Derived Small Extracellular Vesicles Alleviate Diabetic Retinopathy by Delivering miR-22-3p to Inhibit NLRP3 Inflammasome Activation.

PURPOSE: This study aimed to investigate the effect of mesenchymal stem cell (MSC)-derived small extracellular vesicles (sEVs) on diabetic retinopathy (DR) and its underlying mechanism. METHODS: In vivo, MSC-sEVs were injected intravitreally into diabetic rats to determine the therapeutic efficacy. In vitro, MSC-sEVs with/without miR-22-3p inhibition were cocultured with advanced glycation end-products (AGEs)-induced microglia with/without NLRP3 overexpression to explore the molecular mechanism. RESULTS: In vivo, MSC-sEVs inhibited NLRP3 inflammasome activation, suppressed microglial activation, decreased inflammatory cytokines levels in the retina, and alleviated DR as evidenced by improved histological morphology and blood-retinal barrier function. Based on miRNA sequencing of MSC-sEVs, bioinformatic software, and dual-luciferase reporter assay, miR-22-3p stood out as the critical molecule for the role of MSC-sEVs in regulating NLRP3 inflammasome activation. Diabetic rats had lower level of miR-22-3p in their retina than those of control and sEV-treated rats. Confocal microscopy revealed that sEV could be internalized by microglia both in vivo and in vitro. In vitro, compared with sEV, the anti-inflammation effect of sEVmiR-22-3p(-) on AGEs-induced microglia was compromised, as they gave a lower suppression of NLRP3 inflammasome activation and inflammatory cytokines. In addition, NLRP3 overexpression in microglia damped the anti-inflammatory effect of sEV. CONCLUSION: These results indicated that MSC-sEVs alleviated DR via delivering miR-22-3p to inhibit NLRP3 inflammasome activation. Our findings indicate that MSC-sEVs might be a potential therapeutic method for DR.

Value of 18F-FDG-PET/CT radiomics combined with clinical variables in the differential diagnosis of malignant and benign vertebral compression fractures.

BACKGROUND: Vertebral compression fractures (VCFs) are common clinical problems that arise from various reasons. The differential diagnosis of benign and malignant VCFs is challenging. This study was designed to develop and validate a radiomics model to predict benign and malignant VCFs with 18F-fluorodeoxyglucose-positron emission tomography/computed tomography (18F-FDG-PET/CT). RESULTS: Twenty-six features (9 PET features and 17 CT features) and eight clinical variables (age, SUVmax, SUVpeak, SULmax, SULpeak, osteolytic destruction, fracture line, and appendices/posterior vertebrae involvement) were ultimately selected. The area under the curve (AUCs) of the radiomics and clinical-radiomics models were significantly different from that of the clinical model in both the training group (0.986, 0.987 vs. 0.884, p < 0.05) and test group (0.962, 0.948 vs. 0.858, p < 0.05), while there was no significant difference between the radiomics model and clinical-radiomics model (p > 0.05). The accuracies of the radiomics and clinical-radiomics models were 94.0% and 95.0% in the training group and 93.2% and 93.2% in the test group, respectively. The three models all showed good calibration (Hosmer-Lemeshow test, p > 0.05). According to the decision curve analysis (DCA), the radiomics model and clinical-radiomics model exhibited higher overall net benefit than the clinical model. CONCLUSIONS: The PET/CT-based radiomics and clinical-radiomics models showed good performance in distinguishing between malignant and benign VCFs. The radiomics method may be valuable for treatment decision-making.

New insight into circRNAs: characterization, strategies, and biomedical applications.

Circular RNAs (circRNAs) are a class of covalently closed, endogenous ncRNAs. Most circRNAs are derived from exonic or intronic sequences by precursor RNA back-splicing. Advanced high-throughput RNA sequencing and experimental technologies have enabled the extensive identification and characterization of circRNAs, such as novel types of biogenesis, tissue-specific and cell-specific expression patterns, epigenetic regulation, translation potential, localization and metabolism. Increasing evidence has revealed that circRNAs participate in diverse cellular processes, and their dysregulation is involved in the pathogenesis of various diseases, particularly cancer. In this review, we systematically discuss the characterization of circRNAs, databases, challenges for circRNA discovery, new insight into strategies used in circRNA studies and biomedical applications. Although recent studies have advanced the understanding of circRNAs, advanced knowledge and approaches for circRNA annotation, functional characterization and biomedical applications are continuously needed to provide new insights into circRNAs. The emergence of circRNA-based protein translation strategy will be a promising direction in the field of biomedicine.

Circular RNA hsa_circ_001350 contributes to osteosarcoma progression by regulating microRNA-578/CCR4-NOT transcription complex and subunit 7/Wnt signaling.

Accumulating evidence has revealed that circular RNAs (circRNAs) play important roles in cancer by sponging microRNAs (miRNAs). A previous study has shown that hsa_circ_001350 expression is increased in glioma tissue samples and cells and that hsa_circ_001350 directly sponges miR-1236. Here, we investigated the role of hsa_circ_001350 in osteosarcoma (OS). Bioinformatics analysis was performed to examine the potential interactions among hsa_circ_001350, miR-578, and the CCR4-NOT transcription complex and subunit 7 (CNOT7). Reverse transcription-quantitative polymerase chain reaction and western blotting were performed to analyze gene expression and protein levels, respectively. Hsa_circ_001350 expression was upregulated in OS tissues and cell lines. The deletion of hsa_circ_001350 inhibited the proliferation, migration, and invasion of OS cells. The downregulation of hsa_circ_001350 suppressed CNOT7 expression by sponging miR-578 as confirmed by rescue experiments and luciferase reporter assays. Specifically, the depletion of hsa_circ_001350 inhibited the protein expression of ß-catenin, cyclin D1, and c-myc in OS cells, and CNOT7 overexpression reversed this effect. We conclude that hsa_circ_001350 contributes to OS progression by regulating miR-578/CNOT7/Wnt signaling. Thus, hsa_circ_001350, miR-578, and CNOT7 may be potential targets for the treatment of OS.

Clinical and CT Imaging Differences Between Gastric Schwannoma and Gastric Leiomyoma.

OBJECTIVE: To determine whether computed tomography (CT) imaging features can be used to differentiate gastric schwannoma (GS) from gastric leiomyoma (GL) and to develop a nomogram as a predictive model. STUDY DESIGN: Retrospective study. Place and Duration of the Study: Department of Medical Imaging, Affiliated Hospital of Jining Medical University, Jining, Shandong, China, from July 2009 to June 2022. METHODOLOGY: Clinical and imaging data of 43 patients with GS and 57 patients with GL were analysed retrospectively. The independent factors for differentiating GS and GL were obtained by the logistic regression analysis. Receiver operating characteristic curve (ROC) was plotted, area under curve (AUC) and calibration tests were used to evaluate the diagnostic efficiency of the model. RESULTS: The GS group had more females and was older than the GL group (p <0.05). There were statistical differences between the two groups in tumour location, growth mode, LD/SD ratio, necrosis, ulcers, the presence of tumour-associated lymph nodes, enhancement degree, and the HU (Hounsfield units) values of tumour in the venous phase and delayed phase (p <0.05). Logistic regression analysis showed that tumour location, growth mode, LD/SD (long and short diameters) ratio, and the presence of tumour-associated lymph nodes were independent factors in differentiating GS from GL, and a nomogram model was established accordingly. When the model threshold was >0.319, the AUC was 0.987 (95% confidence interval [CI] 0.941~0.999). The sensitivity and specificity were 97.7% and 94.7%, respectively. CONCLUSION: The proposed nomogram model based on CT imaging features can be used to differentiate GS from GL. KEY WORDS: Gastric leiomyoma, Gastric schwannoma, Computed tomography, Diagnosis.

Protective effects of ferulic acid against ionizing radiation-induced oxidative damage in rat lens through activating Nrf2 signal pathway.

AIM: To examine the protection of ferulic acid (FA) against ionizing radiation (IR)-induced lens injury in rats, as well as the underlying mechanisms. METHODS: FA (50 mg/kg) was administered to rats for 4 consecutive days before they were given 10 Gy γ-radiation, as well as for 3 consecutive days afterward. Two weeks after radiation, the eye tissues were collected. Histological alterations were evaluated by hematoxylin-eosin staining. Enzyme linked immunosorbent assay (ELISA) was utilized to assess the activities of glutathione reductase (GR) and superoxide dismutase (SOD), as well as the levels of glutathione (GSH) and malondialdehyde (MDA) in the lenses. The protein and mRNA levels of Bcl-2, caspase-3, Bax, heme oxygenase-1 (HO-1), and glutamate-cysteine ligase catalytic subunit (GCLC) were quantified using Western blot and quantitative reverse transcription polymerase chain reaction, respectively. With nuclear extracts, the nuclear factor erythroid-2 related factor (Nrf2) protein expressions in the nuclei were also measured. RESULTS: Rats exposed to IR showed lens histological alterations which could be alleviated by FA. FA treatment reversed apoptosis-related markers in IR-induced lens, as evidenced by lower levels of Bax and caspase-3 and higher level of Bcl-2. Furthermore, IR induced oxidative damage manifested by decreased GSH level, increased MDA level, and decreased SOD and GR activities. FA boosted nuclear translocation of Nrf2 and increased the expressions of HO-1 and GCLC to inhibit oxidative stress, as evidenced by an increase in GSH, a decrease in MDA, and an increase in GR and SOD activities. CONCLUSION: FA may work well in preventing and treating IR-induced cataract through promoting the Nrf2 signal pathway to attenuate oxidative damage and cell apoptosis.

METTL3 protects METTL14 from STUB1-mediated degradation to maintain m6 A homeostasis.

N6 -Methyladenosine (m6 A) is an important RNA modification catalyzed by methyltransferase-like 3 (METTL3) and METTL14. m6 A homeostasis mediated by the methyltransferase (MTase) complex plays key roles in various biological processes. However, the mechanism underlying METTL14 protein stability and its role in m6 A homeostasis remain elusive. Here, we show that METTL14 stability is regulated by the competitive interaction of METTL3 with the E3 ligase STUB1. STUB1 directly interacts with METTL14 to mediate its ubiquitination at lysine residues K148, K156, and K162 for subsequent degradation, resulting in a significant decrease in total m6 A levels. The amino acid regions 450-454 and 464-480 of METTL3 are essential to promote METTL14 stabilization. Changes in STUB1 expression affect METTL14 protein levels, m6 A modification and tumorigenesis. Collectively, our findings uncover an ubiquitination mechanism controlling METTL14 protein levels to fine-tune m6 A homeostasis. Finally, we present evidence that modulating STUB1 expression to degrade METTL14 could represent a promising therapeutic strategy against cancer.

Amide Proton Transfer Weighted MR Imaging for Predicting Meningioma Stiffness: A Feasibility Study.

BACKGROUND: Stiffness of meningioma is an important factor affecting the surgical resection and the prognosis of patients. PURPOSE: To examine the feasibility of APTw-magnetic resonance imaging (MRI) in evaluating meningioma stiffness. STUDY TYPE: Retrospective. POPULATION: Seventy-one patient with meningiomas, 39 were male and 32 were female; the mean age was 51 ± 10 years. FIELD STRENGTH/SEQUENCE: 3.0T; Turbo-spin-echo T1 -weighted and Gd-T1 -weighted sequence; Turbo-spin-echo T2 -weighted sequence; 2D fat-suppressed, turbo-spin-echo APTw pulse sequence. ASSESSMENT: The T1 WI signal intensity score, T2 WI signal intensity score, APTwmin , APTwmax , and APTwmean values were compared between soft, medium stiff and stiff meningiomas or non-stiff meningiomas and stiff meningiomas group. STATISTICAL TESTS: Chi-square test, one-way ANOVA analysis, independent-samples t-test, intra-class correlation coefficient, rank-sum test, receiver operating characteristic curve analysis. P < 0.05 was considered statistically significant in all tests. RESULTS: APTwmin and APTwmean in the stiff group were significantly lower than that in the non-stiff group (2.79% ± 0.42% vs. 1.90% ± 0.60% and 3.20% ± 0.31% vs. 2.55% ± 0.61%). APTwmin and APTwmean in the stiff group were significantly lower than that in the medium stiff and soft groups (1.90% ± 0.60% vs. 2.69% ± 0.40% and 3.12% ± 0.32%, 2.55% ± 0.61% vs. 3.17% ± 0.33% and 3.39% ± 0.18%), APTwmin in the medium stiff group was significantly lower than in the soft group, there was no significant difference in APTwmean between the medium stiff and soft groups (P = 0.190). APTwmin showed the best diagnostic performance for evaluating meningioma stiffness with an area under the curve of 0.913, when the APTwmin was lower than 2.4%, the meningioma was defined as a stiff tumor, the sensitivity, specificity, and accuracy were 87.1%, 87.5%, and 85.9%, respectively. DATA CONCLUSION: APTw-MRI could be used to evaluate meningioma stiffness, with APTwmin having the best evaluative efficiency. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY: Stage 1.

MYB30 integrates light signals with antioxidant biosynthesis to regulate plant responses during postsubmergence recovery.

Submergence is an abiotic stress that limits agricultural production world-wide. Plants sense oxygen levels during submergence and postsubmergence reoxygenation and modulate their responses. Increasing evidence suggests that completely submerged plants are often exposed to low-light stress, owing to the depth and turbidity of the surrounding water; however, how light availability affects submergence tolerance remains largely unknown. Here, we showed that Arabidopsis thaliana MYB DOMAIN PROTEIN30 (MYB30) is an important transcription factor that integrates light signaling and postsubmergence stress responses. MYB DOMAIN PROTEIN30 protein abundance decreased upon submergence and accumulated during reoxygenation. Under submergence conditions, CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1), a central regulator of light signaling, caused the ubiquitination and degradation of MYB30. In response to desubmergence, however, light-induced MYB30 interacted with MYC2, a master transcription factor involved in jasmonate signaling, and activated the expression of the VITAMIN C DEFECTIVE1 (VTC1) and GLUTATHIONE SYNTHETASE1 (GSH1) gene families to enhance antioxidant biosynthesis. Consistent with this, the myb30 knockout mutant showed increased sensitivity to submergence, which was partially rescued by overexpression of VTC1 or GSH1. Thus, our findings uncover the mechanism by which the COP1-MYB30 module integrates light signals with cellular oxidative homeostasis to coordinate plant responses to postsubmergence stress.