In-situ strategies for melamine-functionalized graphene oxide nanosheets-based nanocomposite proton exchange membranes in wide-temperature range applications.

The traditional preparation of nanocomposite proton exchange membranes (PEM) is hindered by poor organic-inorganic interface compatibility, insufficient proton-conducting sites, easy aggregation of nanoparticles, and difficulty in leveraging nanoscale advantages. In this study, a novel method involving electrochemical anodic oxidation exfoliation was employed to prepare melamine-coated graphene oxide (Me@GO), which was subsequently subjected to in-situ polymerization with poly(2,5-benzimidazole) (ABPBI) to prepare a Me@GO/ABPBI composite proton exchange membrane. Benefiting from the strong hydrogen bonding and large π stacking interactions, melamine (Me) tightly bound to graphene oxide (GO), effectively preventing the secondary aggregation of GO after exfoliation. Moreover, the abundant alkaline functional groups of melamine enhanced the enhancement of phosphoric acid (PA) retention in the Me@GO/ABPBI membranes, thereby increasing the number of proton-conducting sites. The experimental results indicated that the introduction of Me@GO enhanced membrane properties. For Me@GO at a concentration of 1 wt%, the tensile strength of the 1Me@GO/ABPBI composite membrane reached 207 MPa, nearly 2.52 times that of the pure membrane. The proton conductivity of the 1Me@GO/ABPBI composite membrane reached 0.01 S cm-1 across a wide temperature range (40-180 °C), peaking at 0.087 S cm-1 at 180 °C. Additionally, a single-cell incorporating the 1Me@GO/ABPBI composite membrane achieved a peak power density of 0.304 W cm-2 at 160 °C, nearly 1.46 times that of the pure membrane. Benefiting from the well-dispersed and PA-enriched proton channels provided by Me@GO, the Me@GO/ABPBI composite membrane exhibits excellent prospects for wide-temperature range (40-180 °C) applications.

Enhancing the differential diagnosis of small pulmonary nodules: a comprehensive model integrating plasma methylation, protein biomarkers, and LDCT imaging features.

BACKGROUND: Accurate differentiation between malignant and benign pulmonary nodules, especially those measuring 5-10 mm in diameter, continues to pose a significant diagnostic challenge. This study introduces a novel, precise approach by integrating circulating cell-free DNA (cfDNA) methylation patterns, protein profiling, and computed tomography (CT) imaging features to enhance the classification of pulmonary nodules. METHODS: Blood samples were collected from 419 participants diagnosed with pulmonary nodules ranging from 5 to 30 mm in size, before any disease-altering procedures such as treatment or surgical intervention. High-throughput bisulfite sequencing was used to conduct DNA methylation profiling, while protein profiling was performed utilizing the Olink proximity extension assay. The dataset was divided into a training set and an independent test set. The training set included 162 matched cases of benign and malignant nodules, balanced for sex and age. In contrast, the test set consisted of 46 benign and 49 malignant nodules. By effectively integrating both molecular (DNA methylation and protein profiling) and CT imaging parameters, a sophisticated deep learning-based classifier was developed to accurately distinguish between benign and malignant pulmonary nodules. RESULTS: Our results demonstrate that the integrated model is both accurate and robust in distinguishing between benign and malignant pulmonary nodules. It achieved an AUC score 0.925 (sensitivity = 83.7%, specificity = 82.6%) in classifying test set. The performance of the integrated model was significantly higher than that of individual methylation (AUC = 0.799, P = 0.004), protein (AUC = 0.846, P = 0.009), and imaging models (AUC = 0.866, P = 0.01). Importantly, the integrated model achieved a higher AUC of 0.951 (sensitivity = 83.9%, specificity = 89.7%) in 5-10 mm small nodules. These results collectively confirm the accuracy and robustness of our model in detecting malignant nodules from benign ones. CONCLUSIONS: Our study presents a promising noninvasive approach to distinguish the malignancy of pulmonary nodules using multiple molecular and imaging features, which has the potential to assist in clinical decision-making. TRIAL REGISTRATION: This study was registered on ClinicalTrials.gov on 01/01/2020 (NCT05432128). https://classic. CLINICALTRIALS: gov/ct2/show/NCT05432128 .

Efficacy and Safety of Nefecon in Patients with Immunoglobulin A Nephropathy from Mainland China: 2-Year NefIgArd Trial Results.

BACKGROUND: Immunoglobulin A nephropathy (IgAN), an immune-mediated kidney disease, is particularly prevalent among individuals of East Asian ancestry. Nefecon is a novel, oral, targeted-release budesonide formulation designed to inhibit galactose-deficient-IgA1 formation underlying IgAN pathophysiology. We present findings in patients with IgAN from mainland China participating in the 2-year, multicenter, randomized, double-blind, phase 3 NefIgArd trial of Nefecon. METHODS: Patients (aged ≥18 years) with primary IgAN (estimated glomerular filtration rate [eGFR] 35-90 ml/min per 1.73 m2, persistent proteinuria [urine protein-creatinine ratio ≥0.8 g/g or proteinuria ≥1 g/24 h] despite optimized renin-angiotensin system blockade) received Nefecon or placebo over 9 months, followed by a 15-month follow-up phase on supportive care alone. The primary efficacy end point was time-weighted average of eGFR over 2 years. RESULTS: Sixty-two patients from mainland China were included in this prespecified analysis. The primary efficacy end point was 9.6 ml/min per 1.73 m2 (95% confidence interval [CI], 2.0 to 19.8) in favor of Nefecon versus placebo. This was consistent with (and numerically greater than) that of the global study population. Time to confirmed 30% eGFR reduction or kidney failure from baseline was substantially delayed with Nefecon (patients with an event: 9%) versus placebo (30%; hazard ratio, 0.21; 95% CI, 0.04 to 0.73]). No deaths were reported in the China cohort. In the Nefecon group, treatment-emergent serious adverse events were reported by one patient during treatment and two patients during follow-up (versus no patients and seven patients, respectively, in the placebo group). No severe infections requiring hospitalization were reported. CONCLUSIONS: Nefecon treatment for 9 months showed greater preservation of eGFR over 2 years compared with placebo. The efficacy outcomes were consistent with global study results, with a numerically greater treatment benefit observed in patients from China. Nefecon was well tolerated, with no unexpected safety signals.

An Organic Acid-Alkali Coregulated Ionic Liquid Electrolyte Enabling Wide-Temperature-Range Proton Battery.

The broad applications of rechargeable batteries urge people to develop alternative energy storage devices with sustainable resources, high capacity, long cycling life, and wide-temperature operability. Aqueous proton batteries are considered as a state-of-the-art energy storage system due to their intrinsic safety and low cost. However, aqueous electrolytes have a low boiling point and narrow electrochemical stability window, limiting their applications in wide-temperature and high-energy batteries. Herein, a hybrid organic ionic liquid electrolyte with organic alkali 1-methyl-1,2,4-triazole (MTA) protonated by organic acid bis(trifluoromethysulfonyl)imide (HTFSI) as proton carriers and tetramethylene sulfone (TMS) as the solvent, noted as HTFSI-MTA-TMS, exhibited the stable electrochemical windows exceeding 5 V at -20 °C and 3.5 V at 80 °C. Benefiting from this electrolyte, the assembled MnO2-S//MoO3 button proton full battery can display an operation voltage up to 1.8 V, energy density of 44.8 Wh kg-1, and good cycling stability at room temperature when bis(trifluoromethanesulfonyl)imide manganese (II) salt (Mn(TFSI)2) is introduced into the electrolyte, and run well in a wide-temperature range (-20 °C-60 °C). The work reveals the potential of organic acid-alkali coregulated electrolytes to meet the need of energy storage in a wide-temperature range and will advance the development of high-energy proton batteries.

A Molten Alkali Approach to Tailor Hydroxyl Groups of Hexaazatrinaphthalene Toward High-Capacity and Low-Potential Anode of Aqueous Proton Batteries.

Hexaazatrinaphthalene (HATN) has attracted a lot of attention in aqueous proton batteries (APBs). However, its redox potential as an anode is insufficiently negative. The introduction of electron-donating substituent groups, such as hydroxyl groups, is considered as a good approach to reduce the redox potential of HATN. Nevertheless, manufacturing hydroxyl-substituted HATN (HATN-OH) requires either expensive precursors or multi-step process, limiting their research. Herein, a straightforward strategy is proposed to synthesize HATN-OH based on the nucleophilic substitution reaction of halogenated HATN in a molten alkali. The redox potential of 1,2,7,8,13,14-hexahydroxy-5,6,11,12,17,18-hexaazatrinaphthalene (34-HATN-6OH) electrode may be lowered by 0.15 V in comparison to HATN, and exhibits a high specific capacity, low redox potential, remarkable rate capability, and outstanding long-term cycling performance. The electrochemical redox kinetics is significantly enhanced owing to the formation of rapid proton transport channels created by intermolecular hydrogen bond network. The assembled MnO2||34-HATN-6OH full battery delivers a high discharge voltage (1.16 V) and cycling stability (74% capacity retention after 5000 cycles). This study provides a general cost-effective molten alkali approach for the synthesis of hydroxyl-substituted conjugated small molecules from their halogenated counterparts and further enriches the regulation means of electro-chemical performances of organic electrodes for enabling high-capacity and high-voltage APBs.

Prognostic Role of Hemoglobin Combined With Geriatric Nutritional Risk Index in Patients With Vater Ampulla Carcinoma Undergoing Pancreaticoduodenectomy.

OBJECTIVES: The purpose of this investigation was to assess the prognostic importance of the combination of preoperative hemoglobin (Hb) levels and Geriatric Nutritional Risk Index (GNRI) in forecasting postoperative survival outcomes for patients undergoing pancreaticoduodenectomy (PD) due to Vater ampulla carcinoma (VPC). METHODS: The GNRI nutritional screening was conducted for all patients, and patient outcomes, including overall survival (OS), were subsequently monitored. An H- GNRI scoring system was established using the optimal critical values of 125.5 g/L for Hb and 91.72 for GNRI, as determined by X-tile software. According to the H-GNRI score, the patients were categorized into three groups, namely low H-GNRI group (H-GNRI score = 0, n = 47) with Hb < 125.5 g/L and GNRI < 91.72; medium H-GNRI group (H-GNRI score = 1, n = 77) with Hb < 125.5 g/L and GNRI ≥ 91.72, or Hb ≥ 125.5 g/L and GNR < 91.72; and high H-GNRI group (H-GNRI score = 2, n = 51) with Hb ≥ 125.5 g/L and GNRI ≥ 91.72. The Kaplan-Meier analysis and log-rank tests were employed to evaluate the OS rate and compare survival disparities among various groups. Additionally, both univariate and multivariate analyses were conducted utilizing the Cox regression model, with p < 0.05 considered statistically significant. Finally, to evaluate the predictive effectiveness of Hb, GNRI, and H-GNRI, a receiver operating characteristic (ROC) curve was constructed to compare the area under curve (AUC) values. RESULTS: The OS rate was higher in patients with high Hb levels (≥ 125.5 g/L) compared to those with low Hb levels (< 125.5 g/L). Likewise, patients in the high GNRI group (≥ 91.72) exhibited significantly superior OS compared to those in the low GNRI group (< 91.72). Compared with both the medium and low H-GNRI groups, the high H-GNRI group demonstrated a notably higher OS rate. The T stage (HR = 2.523, 95% CI: 1.694-3.757, p < 0.001), N stage (HR = 2.018, 95% CI: 1.255-3.246, p = 0.004), and the H-GNRI score (H-GNRI score of 2 used as the baseline; H-GNRI score of 0: HR = 2.569, 95% CI: 1.499-4.402, p < 0.001; H-GNRI score of 1: HR = 1.835, 95% CI: 1.118-3.014, p = 0.016), after adjusting for gender, were determined to be independent significant predictors affecting the OS of patients with VPC. The AUC of H-GNRI was 0.677, exceeding that of Hb levels (0.631) and GNRI (0.615). CONCLUSIONS: The combination of preoperative Hb levels and GNRI demonstrates superior predictive efficacy for VPC patients undergoing PD, compared with either Hb levels or GNRI score alone. Therefore, the H-GNRI score can be utilized to promptly identify high-risk patients, establish comprehensive nutritional pre-rehabilitation plans through interdisciplinary collaboration, and inform decisions regarding additional adjunctive therapies.

The vaginal metabolomics profile with features of polycystic ovary syndrome: a pilot investigation in China.

Background: Polycystic ovary syndrome (PCOS) is the most common metabolic disorder and reproductive endocrine disease, posing an elevated risk to women of reproductive age. Although metabolism differences in serum, amniotic fluid and urine have been documented in PCOS, there remains a paucity of evidence for vaginal fluid. This study aimed to identify the metabolic characteristics and potential biomarkers of PCOS in Chinese women of reproductive age. Methods: We involved ten newly diagnosed PCOS women who attended gynecology at Zhongda Hospital and matched them with ten healthy controls who conducted health check-up programs at Gulou Maternal and Child Health Center in Nanjing, China from January 1st, 2019 to July 31st, 2020. Non-targeted metabolomics based on ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was applied to differentially screen vaginal metabolites between PCOS group and healthy controls. Principal component analysis (PCA), orthogonal partial least-squares discriminant analysis (OPLS-DA) and enrichment analysis were used to observe differences, search for potential biomarkers and enrich related pathways. Results: Among the 20 participants, a total of 195 different metabolites were detected between PCOS group and healthy control group. PCOS and control groups were effectively separated by vaginal fluid. Lipids and lipid-like molecules constituted the majority of differential metabolites. Notably, dopamine exhibited an increased trend in PCOS group and emerged as the most significant differential metabolite, suggesting its potential as a biomarker for identifying PCOS. The application of UHPLC-MS/MS based vaginal metabolomics methods showed significant differences between PCOS and non-PCOS healthy control groups, especially linoleic acid metabolism disorder. Most differential metabolites were enriched in pathways associated with linoleic acid metabolism, phenylalanine metabolism, tyrosine metabolism, nicotinate and nicotinamide metabolism or arachidonic acid metabolism. Conclusions: In this pilot investigation, significant metabolomics differences could be obtained between PCOS and healthy control groups. For PCOS women of reproductive age, vaginal metabolism is a more economical, convenient and harmless alternative to provide careful personalized health diagnosis and potential targets for therapeutic intervention.

Overloaded Vertebral Body Following Consecutive Three-Level Hybrid Surgery Comparing with Anterior Cervical Discectomy and Fusion.

OBJECTIVE: Based on the varying number and relative positions of cervical disc replacement (CDR) and anterior cervical discectomy and fusion (ACDF) procedures, three-segment hybrid surgery (HS) presents a diverse structural approach. Currently, the potential differential effects of HS with different segment combinations and surgical procedures on overloaded vertebral body (OVB) occurrence remain unexplored. The purpose of this retrospective study is to compare the clinical and radiological outcomes of HS and ACDF in treating cervical degenerative disc disease (CDDD), aiming to provide further insights into OVB. METHODS: This study included patients with three-level CDDD who underwent ACDF or HS at our institution. Eligible patients were divided into three groups: Type I (one-level CDR and two-level ACDF), Type II (two-level CDR and one-level ACDF), and ACDF (three-level ACDF). For radiographic analysis, patients were further divided into the Replacement Segment Group and the Nonreplacement Segment Group based on the presence of replacement segments above and below the OVB. Clinical outcomes were evaluated using visual analog scale (VAS) scores for neck and arm pain, Japanese Orthopedic Association (JOA) scores, and neck disability index (NDI) scores. The cervical radiological parameters assessed included (1) vertebral cross-sectional area (CSA), (2) wedge angle (WA), (3) anterior vertebral height (AH), (4) posterior vertebral height (PH), and (5) Hounsfield unit (HU) values. Statistical methods included paired t-test, ANOVA test, and chi-square test. Independent samples t-test, Mann-Whitney U test, and Wilcoxon signed-rank test were used to compare the differences between two groups according to the results of normal distribution test. RESULTS: A total of 123 patients, evenly distributed among three groups, were included and were well matched in terms of demographic characteristics. The likelihood of vertebral body collapse (VBC) was notably higher in the ACDF group (41.5%) compared with the Type I (17.9%) and Type II (8.9%) groups (p < 0.01). Following surgery, both at 3 and 6 months, the ACDF group demonstrated higher VAS neck scores and NDI scores compared with the Type I and Type II groups (p < 0.01). Additionally, the WA and AH values of the upper and lower adjacent OVB were consistently lower in the ACDF group than in the Type I and Type II groups at 6 and 12 months and at the final follow-up (p < 0.01). Notably, in the Nonreplacement Segment Group, WA significantly decreased at 12 months postoperatively and at the final follow-up compared with the Replacement Segment Group (p < 0.01). CONCLUSIONS: Three levels of HS appear to reduce stress concentrations and alleviate morphological changes in OVB. The occurrence of more VBC patients with OVB was associated with the use of Zero-P or Zero-P VA implants.

[Impact of Climate Change and Human Activities on the Ecological Sensitivity of the Yangtze River Economic Belt].

The increasing climate change and human activities exert their influence on the ecological sensitivity of the region individually and interactively. Therefore, a clear understanding of the impact of climate change and human activities on ecological sensitivity will enhance the resilience of the regional ecological environment and the level of sustainable development. This study took the Yangtze River Economic Belt, the first demonstration zone of China's ecological civilization construction, as the research object. Based on the meteorological, remote sensing, and statistical data of 130 cities in the whole region from 2001 to 2021, an index system of climate change, human activities, and ecological sensitivity was constructed. Response surface methodology （RSM） was used to explore the effects of climate and anthropogenic single factors and interactions on the ecological sensitivity in each region. The results showed that： ① The ecological sensitivity value of the belt fluctuated and rose in time, rising by 2.2% from 2001 to 2021. In terms of space, the overall spatial distribution was "high in the north and low in the south." In 2021, the proportion of severely and extremely sensitive cities in the Yangtze River Economic Belt reached nearly 50%. ② For a single factor, the distribution of the effect of the same factor had certain characteristics： The areas where the single factors of economic development, rainfall, and temperature had a positive impact on the ecological sensitivity were concentrated in the areas with higher or faster economic development, along and south of the Yangtze River. For the interaction factors, the effect of 78.6% of the factors on the ecological sensitivity was negative interaction, and the change of one factor level would change the direction of the effect of the other factor on the regional sensitivity. ③ The comprehensive ecological management area of the Yangtze River Economic Belt was divided based on the ecological sensitivity and climate sensitivity. The governance areas that needed priority improvement were clustered within the three urban agglomerations and their northern adjacent areas, which meant that the ecological sensitivity and climate sensitivity of a city had spillover effects. This study is expected to provide inspiration for the economic zone and even the national and global efforts in the field of regional ecological governance.

Phase-contrast Magnetic Resonance Angiography of Foot at Ultra-high field 5T System: Visualization of Distal Small Vessels and Enhancement by Warm Water Immersion.

BACKGROUND: Ultra-high field strength MR system has been proved to offer improved visualization of the distal intracranial vessels and branches, but its effectiveness on peripheral vasculatures was not investigated. We aim to assess the visualization of lower-extremity vessels using three-dimensional phase contrast MR angiography (3D PC-MRA) at 5T field-strength through the feet with warm water immersion (WWI). METHODS: Participants were prospectively recruited and underwent 3T, 5T 3D PC-MRA on feet with and without WWI (water temperature between 40 to 45 ℃ for a duration of 10minutes). Patients with suspected lower-extremity vessel diseases underwent CTA for lesion identification. Signal-to-noise ratio (SNR), subjective scoring, quantitative vessel segmentation and flow velocity were performed to assess vessel visualization before and after WWI. Friedman's test was conducted to determine statistical significance. RESULTS: Out of thirty participants (mean age, 46.2±5.9; males, 20; lower-extremity vessel disease, 10), 900 vessel segments were available for evaluation. 5T images showed significantly higher scores of image quality and foot vessel visualization than 3T (all P <.05). WWI further improved the visualizing scores (percentage of score 3: 40.2% vs 66.2%, P =.008), SNR (44.27 vs 67.78, P <.001), total branch count (151.92 ± 29.17 vs 225.63 ± 16.76; P <.001), and the flow velocity (0.72 ± 0.03 vs 0.48 ± 0.11cm/s; P <.001). CONCLUSION: 3D PC-MRA at 5T effectively visualizes foot vessels in patients with lower-extremity disease. Furthermore, WWI can significantly enhance the depiction of distal and small vessels.

Effects of different periodontal interventions on the risk of adverse pregnancy outcomes in pregnant women: a systematic review and network meta-analysis of randomized controlled trials.

Background: Periodontal disease is widespread among pregnant women, and it is possible that taking action to improve oral health conditions can make improvements in adverse pregnancy outcomes. Herein, we summarize the recent evidence using a network meta-analysis to assess the effects of different periodontal treatment intervention strategies on the risk of adverse pregnancy outcomes in pregnant women. Materials and methods: Randomized controlled trials were retrieved from PubMed, Web of Science, Embase, and Cochrane Library databases. After literature screening, data extraction, and quality evaluation of the included literature were performed, the R studio 4.2.2 "netmeta" package was used for the network meta-analysis. Results: A total of 20 studies were included, and 5 adverse pregnancy outcomes (preterm birth, low birth weight, preterm birth and/or low birth weight infants, small for gestational age, and pre-eclampsia) were considered to examine the effects of different periodontal treatment interventions strategies on the risk of the abovementioned outcome indicators. The results of the network meta-analysis demonstrated that the three periodontal treatment intervention strategies of sub- and/or supra-gingival scaling and root planing + chlorhexidine rinsing (SRP + CR), sub- and/or supra-gingival scaling and root planing+chlorhexidine rinsing + tooth polishing and plaque control (SRP + CR + TP), and sub- and/or supra-gingival scaling and root planing +sonic toothbrush + tooth polishing and plaque control (SRP + ST + TP) reduced the risk of preterm birth [odds ratio (OR) = 0.29, 95% confidence interval (CI) (0.10-0.88), OR = 0.25, 95CI% (0.10-0.63), OR = 0.28, 95CI% (0.11-0.69), respectively]. In addition, two periodontal treatment intervention strategies, SRP + CR and SRP + CR + TP, were effective methods in terms of the risk of preterm birth and/or low birth weight [OR = 0.18, 95CI% (0.06-0.52), OR = 0.31, 95CI% (0.12-0.79)]. Conclusion: The available evidence suggests that the risk of preterm birth and preterm birth and/or low birth weight can be reduced with certain periodontal treatment intervention strategies. Future studies should focus on optimizing intervention strategies and the optimal timing for different periods of pregnancy, in order to provide a reference for pregnant women's healthcare. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=407901, CRD42023407901.

The synergistic effects of anoikis-related genes and EMT-related genes in the prognostic prediction of Wilms tumor.

Wilms tumor (WT) is the most common type of malignant abdominal tumor in children; it exhibits a high degree of malignancy, grow rapidly, and is prone to metastasis. This study aimed to construct a prognosis model based on anoikis-related genes (ARGs) and epithelial-mesenchymal transition (EMT)-related genes (ERGs) for WT patients; we assessed the characteristics of the tumor microenvironment and treatment efficacy, as well as identifying potential therapeutic targets. To this end, we downloaded transcriptome sequencing data and clinical data for WT and normal renal cortices and used R to construct and validate the prognostic model based on ARGs and ERGs. Additionally, we performed clinical feature analysis, nomogram construction, mutation analysis, drug sensitivity analysis, Connectivity Map (cMAP) analysis, functional enrichment analysis, and immune infiltration analysis. Finally, we screened the hub gene using the STRING database and validated it via experiments. In this way, we constructed a model with good accuracy and robustness, which was composed of seven anoikis- and EMT-related genes. Paclitaxel and mesna were selected as potential chemotherapeutic drugs and adjuvant chemotherapeutic drugs for the WT high-risk group by using the Genomics of Drug Sensitivity in Cancer (GDSC) and cMAP compound libraries, respectively. We proved the existence of a strong correlation between invasive immune cells and prognostic genes and risk scores. Next, we selected NTRK2 as the hub gene, and in vitro experiments confirmed that its inhibition can significantly inhibit the proliferation and migration of tumor cells and promote late apoptosis. In summary, we screened out the potential biomarkers and chemotherapeutic drugs that can improve the prognosis of patients with WT.

Tissue-specific knockout in the Drosophila neuromuscular system reveals ESCRT's role in formation of synapse-derived extracellular vesicles.

Tissue-specific gene knockout by CRISPR/Cas9 is a powerful approach for characterizing gene functions during development. However, this approach has not been successfully applied to most Drosophila tissues, including the Drosophila neuromuscular junction (NMJ). To expand tissue-specific CRISPR to this powerful model system, here we present a CRISPR-mediated tissue-restricted mutagenesis (CRISPR-TRiM) toolkit for knocking out genes in motoneurons, muscles, and glial cells. We validated the efficacy of CRISPR-TRiM by knocking out multiple genes in each tissue, demonstrated its orthogonal use with the Gal4/UAS binary expression system, and showed simultaneous knockout of multiple redundant genes. We used CRISPR-TRiM to discover an essential role for SNARE components in NMJ maintenance. Furthermore, we demonstrate that the canonical ESCRT pathway suppresses NMJ bouton growth by downregulating retrograde Gbb signaling. Lastly, we found that axon termini of motoneurons rely on ESCRT-mediated intra-axonal membrane trafficking to release extracellular vesicles at the NMJ. Thus, we have successfully developed an NMJ CRISPR mutagenesis approach which we used to reveal genes important for NMJ structural plasticity.

Identification of a cellular role of hemolysin co-regulatory protein (Hcp) in Vibrio alginolyticus modulating substrate metabolism and biofilm formation by cAMP-CRP.

Cyclic AMP (cAMP) and cAMP receptor protein (CRP) system controls catabolic enzyme expression based on metabolite concentrations in bacteria. Hemolysin co-regulatory protein (Hcp) is well known as a molecular chaperone for virulence factor secretion of the type VI secretion system (T6SS). However, the intracellular role of Hcp involving in bacterial physiological processes remains unknown. To clarify that, we constructed a single hcp mutant strain and analyzed their effects on the physiological processes of Vibrio alginolyticus. The omics results revealed the extensive involvement of Hcp in the catabolic metabolism in bacteria. Simultaneously, Hcp1 and Hcp2 played opposing regulatory roles on the bacterial growth, biofilm formation, and intracellular cAMP-CRP levels during cultivation in a glucose medium. Furthermore, the interacting protein screening and co-immunoprecipitation (Co-IP) assays confirmed that the glucose-specific phosphoenolpyruvate (PEP)-phosphotransferase system (PTS) enzyme IIA component (EIIAglc) was a key interacting partner with Hcp proteins as well as class I adenylyl cyclase (AC-I) in Vibrio alginolyticus. These results indicated that, to achieve cellular homeostasis, Hcp1 and Hcp2 might exert antagonistic and synergistic effects, respectively, on the interaction between EIIAglc and AC thus cooperatively regulating intracellular cAMP-CRP production.

Intracellular osteopontin potentiates the immunosuppressive activity of mesenchymal stromal cells.

INTRODUCTION: Mesenchymal stromal cell (MSC)-based cell therapy is a promising approach for various inflammatory disorders based on their immunosuppressive capacity. Osteopontin (OPN) regulates several cellular functions including tissue repair, bone metabolism and immune reaction. However, the biological function of OPN in regulating the immunosuppressive capacity of MSCs remains elusive. OBJECTIVES: This study aims to highlight the underlying mechanism of the proinflammatory cytokines affect the therapeutic ability of MSCs through OPN. METHODS: MSCs in response to the proinflammatory cytokines were collected to determine the expression profile of OPN. In vitro T-cell proliferation assays and gene editing were performed to check the role and mechanisms of OPN in regulating the immunosuppressive capacity of MSCs. Inflammatory disease mouse models were established to evaluate the effect of OPN on improving MSC-based immunotherapy. RESULTS: We observed that OPN, including its two isoforms iOPN and sOPN, was downregulated in MSCs upon proinflammatory cytokine stimulation. Interestingly, iOPN, but not sOPN, greatly enhanced the immunosuppressive activity of MSCs on T-cell proliferation and thus alleviated the inflammatory pathologies of hepatitis and colitis. Mechanistically, iOPN interacted with STAT1 and mediated its deubiquitination, thereby inducing the master immunosuppressive mediator inducible nitric oxide synthase (iNOS) in MSCs. In addition, iOPN expression was directly downregulated by activated STAT1, which formed a negative feedback loop to restrain MSC immunosuppressive capacity. CONCLUSION: Our findings demonstrated that iOPN expression modulation in MSCs is a novel strategy to improve MSC-based immunotherapy.

Fisher discriminant analysis of multimodal ultrasound in diagnosis of cervical metastatic lymph nodes in papillary thyroid cancer.

Background/Aims: The purpose of this study was to develop a diagnostic model utilizing multimodal ultrasound parameters to aid in the detection of cervical lymph node metastasis in papillary thyroid cancer (PTC) patients. Methods: The study included 84 suspicious lymph nodes from 69 PTC patients, all of whom underwent fine needle aspiration with pathological results. Data from conventional grayscale ultrasound, shear wave elastography (SWE), and superb microvascular imaging were analyzed. Key ultrasound features were compared between benign and metastatic groups to create a diagnostic model using Fisher's stepwise discriminant analysis. The model's effectiveness was assessed with self-testing, cross-validation, and receiver operating characteristic curve analysis. Results: Four features, namely lymphatic hilum (X1), cortical hyperechogenicity (X2), vascular pattern (X4), and SWEmean (X7), were integral to the discriminant analysis, resulting in the equation: Y1 = -3.461 + 2.423X1 + 0.321X2 + 1.620X4 + 0.109X7, Y2 = -8.053 + 0.414X1 + 2.600X2 + 2.504X4 + 0.192X7. If Y1 < Y2, the LN would be diagnosed as metastatic lymph nodes. The model demonstrated an area under the curve of 0.833, with a sensitivity of 83.33% and specificity of 83.33%. Conclusions: The multimodal ultrasound diagnostic model, established through Fisher's stepwise discriminant analysis, proved effective in identifying metastatic lymph nodes in PTC patients.

Progress in Transdifferentiation of Autologous Alternative Cell Sources into Corneal Epithelial Cells.

Corneal limbal epithelial stem cells (LESCs) play a crucial role in corneal epithelium regeneration. Severe damage to these cells can result in limbal stem cell deficiency (LSCD), characterized by repeated corneal conjunctivalization, leading to corneal turbidity and scar formation. Restoring functional LESCs and their ecological location are essential for treating LSCD. The goal of this review is to provide researchers and clinicians with key insights into LESCs biology and to conclude the current cell-based therapies advancement in LSCD treatments. Therapeutic cell resources mainly include mesenchymal stem cells (MSCs), embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), skin keratinocyte stem cells (SKCs), and oral mucosal epithelial cells (OMECs).

Daucosterol alleviates heart failure with preserved ejection fraction through activating PPARα pathway.

Heart failure with preserved ejection fraction (HFpEF) has been increasing in the population in recent years and is mainly characterized by preserved left ventricle ejection fraction (LVEF), diastolic dysfunction and systemic inflammation. Daucosterol (DAU), a glycoside of ß-sitosterol, has good anti-inflammatory and antioxidative properties; however, its effects and mechanisms in HFpEF have not been investigated. To detect whether DAU could alleviate HFpEF, C57BL/6J male mice were fed with N-nitro-l-arginine methyl ester (L-NAME) in drinking water and high fat diet (HFD) and treated with DAU by gavage (i.g.) for 10 weeks. The results showed that DAU treatment significantly alleviated HFpEF in mice. Mechanistically, by controlling PPARα and preventing NF-κB phosphorylation, DAU reduced oxidative stress and the inflammatory response. In conclusion, our study provides a new clue for natural product DAU in alleviating HFpEF.

Network Pharmacology Analysis and Retrospective Cohort Study Revealing the Effect of Qingyi Decoction on Intestinal Paralysis in Patients with Severe Acute Pancreatitis.

BACKGROUND: Intestinal dysfunction plays an important role in the clinical progress and prognosis of severe acute pancreatitis (SAP). Qingyi decoction (QYD) has shown beneficial effects on intestinal function recovery, but the prevention actions of the QYD on intestinal paralysis and its mechanism have not been fully explored. METHODS: The possible molecular mechanism was unraveled by network pharmacology, including active ingredients and potential target prediction, as well as GO, KEGG, and REATCOME pathway enrichment analyses. The potential interactions between the main active ingredients of the QYD and core genes were explored by molecular docking. A retrospective cohort study on 137 patients with SAP from Tianjin Nankai Hospital was conducted to evaluate the preventive effect of QYD on intestinal paralysis. RESULTS: A total of 110 active ingredients in QYD were screened out, and 37 key targets were predicted by network pharmacology. GO, KEGG, and REATCOME enrichment analyses showed that bioinformatics annotation of the hub genes was mainly involved in intestinal epithelial functions and inflammatory response pathways. The main components of QYD possessed good affinity with IL-6, TNF, CASP3, CXCL8, and CRP by molecular docking. Patients who used QYD plus usual care seemed to have fewer intestinal paralysis rates, lower risk of renal insufficiency, ARDS and blood purification therapy, and shorter hospital and ICU stays. The multivariable regression analyses indicated that the mode of nasogastric and enemas administration of QYD (P = 0.010) and timely intervention with QYD (P = 0.045) were the independent protective factors for intestinal paralysis prevention in patients with SAP. CONCLUSION: In conclusion, QYD can be used as an effective adjuvant procedure to prevent the occurrence and development of intestinal paralysis in patients with SAP. The mechanisms may be involved in the anti-inflammatory response and maintenance of intestinal epithelial function.