MLH1 Inhibits Metastatic Potential of Pancreatic Ductal Adenocarcinoma via Downregulation of GPRC5C.

DNA mismatch repair gene MutL homolog-1 (MLH1) has divergent effects in many cancers, however, its impact on the metastasis of pancreatic ductal adenocarcinoma (PDAC) remains unclear. In this study, MLH1 stably overexpressed (OE) and knockdowned (KD) sub-lines were established. Wound-healing and Transwell assays were used to evaluate cell migration/invasion. In vivo metastasis was investigated in orthotopic implantation models (SCID mice). RT-qPCR and western blotting were adopted to show gene/protein expression. MLH1 down-stream genes were screened by transcriptome sequencing. Tissue microarray-based immunohistochemistry was applied to determine protein expression in human specimens. In successfully generated sub-lines, OE cells presented weaker migration/invasion abilities, compared with controls, while in KD cells these abilities were significantly stronger. The metastasis-inhibitory effect of MLH1 was also observed in mice. Mechanistically, G-protein coupled receptor C5C (GPRC5C) was a key down-stream gene of MLH1 in PDAC cells. Subsequently, transient GPRC5C silencing effectively inhibited cell migration/invasion, and remarkably reversed the pro-invasive effect of MLH1 knockdown in KD cells. In animal models and human PDAC tissues, tumoral GPRC5C expression, negatively associated with MLH1 expressions, was positively correlated with histological grade, vessel invasion, and poor cancer-specific survival. In conclusion, MLH1 inhibits the metastatic potential of PDAC via down-regulation of GPRC5C.

Simultaneously improving the activity and thermostability of hyperthermophillic pullulanase by modifying the active-site tunnel and surface lysine.

Pullulanases are important starch-debranching enzymes that mainly hydrolyze the α-1,6-glycosidic linkages in pullulan, starch, and oligosaccharides. Nevertheless, their practical applications are constrained because of their poor activity and low thermostability. Moreover, the trade-off between activity and thermostability makes it challenging to simultaneously improve them. In this study, an engineered pullulanase was developed through reshaping the active-site tunnel and engineering the surface lysine residues using the pullulanase from Pyrococcus yayanosii CH1 (PulPY2). The specific activity of the engineered pullulanase was increased 3.1-fold, and thermostability was enhanced 1.8-fold. Moreover, the engineered pullulanase exhibited 11.4-fold improvement in catalytic efficiency (kcat/Km). Molecular dynamics simulations demonstrated an anti-correlated movement around the entrance of active-site tunnel and stronger interactions between the surface residues in the engineered pullulanase, which would be beneficial to the activity and thermostability improvement, respectively. The strategies used in this study and dynamic evidence for insight into enzyme performance improvement may provide guidance for the activity and thermostability engineering of other enzymes.

Structural and functional insights into the 2'-O-methyltransferase of SARS-CoV-2.

A unique feature of coronaviruses is their utilization of self-encoded nonstructural protein 16 (nsp16), 2'-O-methyltransferase (2'-O-MTase), to cap their RNAs through ribose 2'-O-methylation modification. This process is crucial for maintaining viral genome stability, facilitating efficient translation, and enabling immune escape. Despite considerable advances in the ultrastructure of SARS-CoV-2 nsp16/nsp10, insights into its molecular mechanism have so far been limited. In this study, we systematically characterized the 2'-O-MTase activity of nsp16 in SARS-CoV-2, focusing on its dependence on nsp10 stimulation. We observed cross-reactivity between nsp16 and nsp10 in various coronaviruses due to a conserved interaction interface. However, a single residue substitution (K58T) in SARS-CoV-2 nsp10 restricted the functional activation of MERS-CoV nsp16. Furthermore, the cofactor nsp10 effectively enhanced the binding of nsp16 to the substrate RNA and the methyl donor S-adenosyl-L-methionine (SAM). Mechanistically, His-80, Lys-93, and Gly-94 of nsp10 interacted with Asp-102, Ser-105, and Asp-106 of nsp16, respectively, thereby effectively stabilizing the SAM binding pocket. Lys-43 of nsp10 interacted with Lys-38 and Gly-39 of nsp16 to dynamically regulate the RNA binding pocket and facilitate precise binding of RNA to the nsp16/nsp10 complex. By assessing the conformational epitopes of nsp16/nsp10 complex, we further determined the critical residues involved in 2'-O-MTase activity. Additionally, we utilize an in vitro biochemical platform to screen potential inhibitors targeting 2'-O-MTase activity. Overall, our results significantly enhance the understanding of viral 2'-O methylation process and mechanism, providing valuable targets for antiviral drug development.

Revealing the effect of sea buckthorn oil, fish oil and structured lipid on intestinal microbiota, colonic short chain fatty acid composition and serum lipid profiles in vivo.

In this study, the effects of sea buckthorn oil (SBO), fish oil (FO) and an enzymatically synthesized structured lipid (SL) on serum, short-chain fatty acids (SCFAs) and intestinal microbiota in Sprague-Dawley (SD) rats were investigated. The results demonstrated that FO, SBO, and SL effectively reduced the levels of high-density lipoprotein cholesterol and low-density lipoprotein cholesterol in the serum of SD rats. SBO increased serum triglyceride levels, while FO elevated total cholesterol levels. Furthermore, all three dietary lipids decreased short-chain fatty acid production and enhanced intestinal microbiota diversity. FO increased the abundance of intestinal microbiota including Romboutsia, Lactobacillus, Escherichia-Shigella, and Lachnospiraceae_NK4A136_group. Conversely, all three dietary lipids reduced the abundance of Klebsiella and Blautia. These findings provide a foundation for understanding the functionality of SBO and FO as well as their potential application in synthesizing novel SLs to regulate intestinal microbiota.

The Anchor of Traditional Culture and Medical Students' Knowledge, Attitudes, and Willingness Towards Organ Donation during Internships: A Cross-Sectional Descriptive Study.

OBJECTIVES: The purpose of this study was to understand the knowledge, attitudes, and willingness of medical students in their clinical internships towards organ donation and to assess how traditional oriental culture plays a role. METHODS: This study conducted an online cross-sectional survey using a cell phone application with a self-designed questionnaire based on the Delphi method. This study was conducted among medical students in their internships in hospitals in nearly 20 provinces of China. The questionnaire consisted of 17 main questions and a knowledge quiz. The results were analyzed with descriptive statistics using the software Microsoft Excel 2021 and SPSS 26. RESULTS: A total of 225 medical students participated in this study, distributed among college, undergraduate, and postgraduate clinical internships. 96% of the medical students agreed with the value of organ donation, but the highest willingness to donate the whole body was only 38.22%. Among them, traditional culture, family values, and religious factors played important both positive and negative roles. CONCLUSION: Medical students have better knowledge about organ donation, but it rarely comes from professional education. Traditional concepts, family values, and religious factors play both a facilitating and hindering role, which can easily lead to moral dilemmas. Although it is not possible to make productive changes in the short term, we could still make improvements through educational changes, research breakthroughs, and institutional improvements.

Current state and influencing factors in airbag management among emergency department nurses: A multicenter study.

BACKGROUND: The emergency department (ED) plays a critical role in establishing artificial airways and implementing mechanical ventilation. Managing airbags in the ED presents a prime opportunity to mitigate the risk of ventilator-associated pneumonia. Nonetheless, existing research has largely overlooked the understanding, beliefs, and practical dimensions of airway airbag management among ED nurses, with a predominant focus on intensive care unit nurses. AIM: To investigate the current status of ED nurses' knowledge, beliefs, and practical behaviors in airway airbag management and their influencing factors. METHODS: A survey was conducted from July 10th to August 10th, 2023, using convenience sampling on 520 ED nurses from 15 tertiary hospitals and 5 sary hospitals in Shanghai. Pathway analysis was utilized to analyze the influencing factors. RESULTS: The scores for ED nurses' airway airbag management knowledge were 60.26 ± 23.00, belief was 88.65 ± 13.36, and behavior was 75.10 ± 19.84. The main influencing factors of airbag management knowledge included participation in specialized nurse or mechanical ventilation training, department, and work experience in the department. Influencing factors of airbag management belief comprised knowledge, department, and participation in specialized nurse or mechanical ventilation training. Primary influencing factors of airbag management behavior included knowledge, belief, department, participation in specialized nurse or mechanical ventilation training, and professional title. The belief in airbag management among ED nurses acted as a partial mediator between knowledge and behavior, with a total effect value of 0.513, and an indirect effect of 0.085, constituting 16.6% of the total effect. CONCLUSION: ED nurses exhibit a positive attitude toward airbag management with relatively standardized practices, yet there remains room for improvement in their knowledge levels. Nursing managers should implement interventions tailored to the characteristics of ED nurses' airbag management knowledge, beliefs, and practices to enhance their airbag management proficiency.

Metabolomics analysis of rice fermented by medicinal fungi providing insights into the preparation of functional food.

Rice, a primary staple food, may be improved in value via fermentation. Here, ten medicinal basidiomycetous fungi were separately applied for rice fermentation. After preliminary screening, Ganoderma boninense, Phylloporia pulla, Sanghuangporus sanghuang and Sanghuangporus weigelae were selected for further LC-MS based determination of the changes in metabolic profile after their fermentation with rice, and a total of 261, 296, 312, and 355 differential compounds were identified, respectively. Most of these compounds were up-regulated and involved in the metabolic pathways of amino acid metabolism, lipid metabolism, carbohydrate metabolism and the biosynthesis of other secondary metabolites. Sanghuangporus weigelae endowed the rice with the highest nutritional and bioactive values. The metabolic network of the identified differential compounds in rice fermented by S. weigelae illustrated their close relationships. In summary, this study provides insights into the preparation and application of potential functional food via the fermentation of rice with medicinal fungi.

Reversible splenial lesion syndrome in Anti-N-methyl-D-aspartate receptor encephalitis.

Here we described an 18-year-old woman who were initially misdiagnosed as psychiatric disorders in a psychiatric institution. She was transferred to our neurological ward because of impaired consciousness. Neuronal antibody testing confirmed the diagnosis of anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis. Cerebral magnetic resonance imaging (MRI) revealed a concomitant disorder named reversible splenial lesion syndrome (RESLES). After administration of combined immunotherapy, the patient recovered completely 3 months after discharge. To our knowledge, co-occurrence of RESLES and anti-NMDAR encephalitis was only described in two patients with teratoma and we provide another case without teratoma. We highlight that anti-NMDAR antibodies can be added to the multiple causes of RESLES. It is therefore imperative for clinicians to detect anti-neuronal antibodies in patients with RESLES to avoid missed diagnosis.

Boosting few-shot rare skin disease classification via self-supervision and distribution calibration.

Due to the difficulty in obtaining clinical samples and the high cost of labeling, rare skin diseases are characterized by data scarcity, making training deep neural networks for classification challenging. In recent years, few-shot learning has emerged as a promising solution, enabling models to recognize unseen disease classes by limited labeled samples. However, most existing methods ignored the fine-grained nature of rare skin diseases, resulting in poor performance when generalizing to highly similar classes. Moreover, the distributions learned from limited labeled data are biased, severely impairing the model's generalizability. This paper proposes a self-supervision distribution calibration network (SS-DCN) to address the above issues. Specifically, SS-DCN adopts a multi-task learning framework during pre-training. By introducing self-supervised tasks to aid in supervised learning, the model can learn more discriminative and transferable visual representations. Furthermore, SS-DCN applied an enhanced distribution calibration (EDC) strategy, which utilizes the statistics of base classes with sufficient samples to calibrate the bias distribution of novel classes with few-shot samples. By generating more samples from the calibrated distribution, EDC can provide sufficient supervision for subsequent classifier training. The proposed method is evaluated on three public skin disease datasets(i.e., ISIC2018, Derm7pt, and SD198), achieving significant performance improvements over state-of-the-art methods.

Correlation of functional magnetic resonance imaging features of primary central nervous system lymphoma with vasculogenic mimicry and reticular fibers.

Objective: To deepen the imaging-pathological mechanism of primary central nervous system lymphoma (PCNSL) and provide a theoretical basis for clinical diagnosis and treatment, the functional magnetic resonance imaging (fMRI) characteristics of PCNSL were analyzed, and the relationship between the fMRI characteristics and vasculogenic mimicry (VM) and reticular fiber in PCNSL was discussed. Methods: Ninety-six patients with PCNSL treated in our hospital were divided into three groups according to the pathological examination results, including strong positive group of VM (n = 40), weak positive group of VM (n = 56), strong positive group of reticular fiber (n = 45) and weak positive group of reticular fiber (n = 51). The levels of augmentation index and apparent diffusion coefficient (ADC) were compared among the groups. receiver operator characteristic (ROC) curve analysis was used to analyze the clinical value of ADC value in differential diagnosis of PCNSL. Results: The levels of augmentation index in the strong positive group of VM were significantly higher than that in the weak positive group of VM, and the ADC value in the strong positive group of VM was significantly lower than that in the weak positive group of VM (P < 0.001). The levels of augmentation index in the strong positive group of reticular fiber were significantly higher than that in the weak positive group of reticular fiber, and ADC value in the strong positive group of reticular fiber was significantly lower than that in reticular fiber weak positive group (P < 0.001). Pearson correlation analysis showed that the levels of augmentation index were positively correlated with VM and reticular fiber (r = 0.529, 0.548, P < 0.001) and the ADC value was negatively correlated with VM and reticular fiber (r = -0.485, -0.513, P < 0.001). There was a significant negative correlation between necrotic lesions and VM (r = -0.185, P < 0.05). The area under the curve (AUC) values of average ADC value, minimum ADC value, and maximum ADC value for individual differential diagnosis of PCNSL were 0.920, 0.901, and 0.702, while the AUC of the combined differential diagnosis was 0.985, with a sensitivity of 95.00 % and a specificity of 92.70 %. Conclusion: The levels of augmentation index and the ADC value of PCNSL focus are significantly correlated with VM and reticular fiber, and there is a strong negative correlation between necrotic lesions and VM. MRI imaging technology is of great significance in revealing the biological behavior of PCNSL, which can effectively reveal the relationship between VM and reticular fibers and the MRI characteristics in PCNSL, thereby providing a new imaging basis for the clinical diagnosis and treatment of PCNSL.

Effectiveness of cardioneuroablation in different subtypes of vasovagal syncope.

BACKGROUND: Cardioneuroablation (CNA) has shown encouraging results in patients with vasovagal syncope (VVS). However, data on different subtypes was scarce. METHODS: This observational study retrospectively enrolled 141 patients [mean age: 40 ± 18 years, 51 males (36.2%)] with the diagnosis of VVS. The characteristics among different types of VVS and the outcomes after CNA were analyzed. RESULTS: After a mean follow-up of 4.3 ± 1.5 years, 41 patients (29.1%) experienced syncope/pre-syncope events after CNA. Syncope/pre-syncope recurrence significantly differed in each subtype (P = 0.04). The cardioinhibitory type of VVS had the lowest recurrence rate after the procedure (n = 6, 16.7%), followed by mixed (n = 26, 30.6%) and vasodepressive (n = 9, 45.0%). Additionally, a significant difference was observed in the analyses of the Kaplan-Meier survival curve (P = 0.02). Syncope/pre-syncope burden was significantly reduced after CNA in the vasodepressive type (P < 0.01). Vasodepressive types with recurrent syncope/pre-syncope after CNA have a lower baseline deceleration capacity (DC) level than those without (7.4 ± 1.0 ms vs. 9.0 ± 1.6 ms, P = 0.01). Patients with DC < 8.4 ms had an 8.1 (HR = 8.1, 95% CI: 2.2-30.0, P = 0.02) times risk of syncope/pre-syncope recurrence after CNA compared to patients with DC ≥ 8.4 ms, and this association still existed after adjusting for age and sex (HR = 8.1, 95% CI: 2.2-30.1, P = 0.02). CONCLUSIONS: Different subtypes exhibit different event-free rates. The vasodepressive type exhibited the lowest event-free rate, but those patients with DC ≥ 8.4 ms might benefit from CNA.

Identification of an ANCA-Associated Vasculitis Cohort Using Deep Learning and Electronic Health Records.

Background: ANCA-associated vasculitis (AAV) is a rare but serious disease. Traditional case-identification methods using claims data can be time-intensive and may miss important subgroups. We hypothesized that a deep learning model analyzing electronic health records (EHR) can more accurately identify AAV cases. Methods: We examined the Mass General Brigham (MGB) repository of clinical documentation from 12/1/1979 to 5/11/2021, using expert-curated keywords and ICD codes to identify a large cohort of potential AAV cases. Three labeled datasets (I, II, III) were created, each containing note sections. We trained and evaluated a range of machine learning and deep learning algorithms for note-level classification, using metrics like positive predictive value (PPV), sensitivity, F-score, area under the receiver operating characteristic curve (AUROC), and area under the precision and recall curve (AUPRC). The deep learning model was further evaluated for its ability to classify AAV cases at the patient-level, compared with rule-based algorithms in 2,000 randomly chosen samples. Results: Datasets I, II, and III comprised 6,000, 3,008, and 7,500 note sections, respectively. Deep learning achieved the highest AUROC in all three datasets, with scores of 0.983, 0.991, and 0.991. The deep learning approach also had among the highest PPVs across the three datasets (0.941, 0.954, and 0.800, respectively). In a test cohort of 2,000 cases, the deep learning model achieved a PPV of 0.262 and an estimated sensitivity of 0.975. Compared to the best rule-based algorithm, the deep learning model identified six additional AAV cases, representing 13% of the total. Conclusion: The deep learning model effectively classifies clinical note sections for AAV diagnosis. Its application to EHR notes can potentially uncover additional cases missed by traditional rule-based methods.

Effectiveness and safety of finerenone in Chinese CKD patients without diabetes: a retrospective, real-world study.

BACKGROUND: Finerenone, a non-steroidal mineralocorticoid receptor antagonist, has previously demonstrated its efficacy and safety in chronic kidney disease (CKD) associated with diabetes mellitus. Given its therapeutic potential, finerenone has been preliminarily explored in clinical practice for non-diabetic CKD patients. The effectiveness and safety in this population require further investigation in a real-world setting. METHODS: This retrospective, real-world analysis included non-diabetic CKD patients receiving finerenone. The main clinical outcomes assessed were changes in urinary albumin-to-creatinine ratio (UACR) and estimated glomerular filtration rate (eGFR). Serum potassium (sK+) levels were also monitored. Data were collected at baseline, and then at 1 month and 3 months following treatment initiation. RESULTS: Totally, 16 patients were included. There was a notable decrease in UACR from 1-month post-treatment, with a further reduction at 3 months, resulting in a median reduction of 200.41 mg/g (IQR, 84.04-1057.10 mg/g; P = 0.028; percent change, 44.52% [IQR, 31.79-65.42%]). The average eGFR at baseline was 80.16 ml/min/1.73m2, with no significant change after 1 month (80.72 ml/min/1.73m2, P = 0.594) and a slight numerical increase to 83.45 ml/min/1.73m2 (P = 0.484) after 3 months. During the 3-month follow-up, sK+ levels showed only minor fluctuations, with no significant differences compared to baseline, and remained within the normal range throughout the treatment period. No treatment discontinuation or hospitalization due to hyperkalemia was observed. CONCLUSION: In non-diabetic CKD patients, finerenone showed good effectiveness and safety within a 3-month follow-up period. This study provides valuable real-world evidence supporting the use of finerenone in non-diabetic CKD and highlights the need for future large-scale prospective research to further validate its efficacy.

Fabrication and Characterization of Docosahexaenoic Acid Algal Oil Pickering Emulsions Stabilized Using the Whey Protein Isolate-High-Methoxyl Pectin Complex.

In this study, the whey protein isolate-high-methoxyl pectin (WPI-HMP) complex prepared by electrostatic interaction was utilized as an emulsifier in the preparation of docosahexaenoic acid (DHA) algal oils in order to improve their physicochemical properties and oxidation stability. The results showed that the emulsions stabilized using the WPI-HMP complex across varying oil-phase volume fractions (30-70%) exhibited consistent particle size and enhanced stability compared to emulsions stabilized solely using WPI or HMP at different ionic concentrations and heating temperatures. Furthermore, DHA algal oil emulsions stabilized using the WPI-HMP complex also showed superior storage stability, as they exhibited no discernible emulsification or oil droplet overflow and the particle size variation remained relatively minor throughout the storage at 25 °C for 30 days. The accelerated oxidation of the emulsions was assessed by measuring the rate of DHA loss, lipid hydroperoxide levels, and malondialdehyde levels. Emulsions stabilized using the WPI-HMP complex exhibited a lower rate of DHA loss and reduced levels of lipid hydroperoxides and malondialdehyde. This indicated that WPI-HMP-stabilized Pickering emulsions exhibit a greater rate of DHA retention. The excellent stability of these emulsions could prove valuable in food processing for DHA nutritional enhancement.

Population Pharmacokinetics and Exposure-Response Relationship of Zimberelimab in Chinese Patients with Advanced Tumors.

This study aimed to establish a population pharmacokinetic (PopPK) model using data from 2 clinical trials of zimberelimab, evaluate the pharmacokinetics (PKs) of zimberelimab, explore the feasibility of 360 mg once every 3 weeks (Q3W) and 480 mg once every 4 weeks (Q4W) as alternative dosage regimens, and analyze the exposure-response relationship of the efficacy and safety of zimberelimab for advanced tumors. The PKs of zimberelimab were described using the 2-compartment model with time-dependent nonlinear elimination. The prediction-corrected visual predictive check was used to evaluate the model's predictive value on blood drug concentrations. In total, 2165 PK observations from 321 participants were included. The PopPK model demonstrated a high level of concordance between the observed data and the predicted values, indicative of a robust fit to the PK data of zimberelimab. The PK variables were similar for the 240 mg once every 2 weeks, 360 mg Q3W, and 480 mg Q4W regimens. No covariates significantly affecting the PK variables in the final model were found. The exposure variables of zimberelimab have no obvious correlations with efficacy and safety, and 360 mg Q3W and 480 mg Q4W are worthy of further study. This study establishes a PopPK model and analyzes the exposure-response relationship of zimberelimab, which helps to explore the potential for alternative dosing regimens and offers a foundation for optimizing therapeutic strategies for advanced cancer patients through simulation-based methods.

Unmasking Bias in AI: A Systematic Review of Bias Detection and Mitigation Strategies in Electronic Health Record-based Models.

OBJECTIVES: Leveraging artificial intelligence (AI) in conjunction with electronic health records (EHRs) holds transformative potential to improve healthcare. Yet, addressing bias in AI, which risks worsening healthcare disparities, cannot be overlooked. This study reviews methods to detect and mitigate diverse forms of bias in AI models developed using EHR data. METHODS: We conducted a systematic review following the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines, analyzing articles from PubMed, Web of Science, and IEEE published between January 1, 2010, and Dec 17, 2023. The review identified key biases, outlined strategies for detecting and mitigating bias throughout the AI model development process, and analyzed metrics for bias assessment. RESULTS: Of the 450 articles retrieved, 20 met our criteria, revealing six major bias types: algorithmic, confounding, implicit, measurement, selection, and temporal. The AI models were primarily developed for predictive tasks in healthcare settings. Four studies concentrated on the detection of implicit and algorithmic biases employing fairness metrics like statistical parity, equal opportunity, and predictive equity. Sixty proposed various strategies for mitigating biases, especially targeting implicit and selection biases. These strategies, evaluated through both performance (e.g., accuracy, AUROC) and fairness metrics, predominantly involved data collection and preprocessing techniques like resampling, reweighting, and transformation. DISCUSSION: This review highlights the varied and evolving nature of strategies to address bias in EHR-based AI models, emphasizing the urgent needs for the establishment of standardized, generalizable, and interpretable methodologies to foster the creation of ethical AI systems that promote fairness and equity in healthcare.

CircSMAD3 represses SMAD3 phosphorylation and ameliorates cardiac remodeling by recruiting YBX1.

Circular RNA (circRNA) has emerged as potential therapeutic targets for cardiovascular diseases. Given the central role of the TGFß signaling pathway in cardiac remodeling and its potential as a therapeutic target, we hypothesized that a circRNA from this pathway could modulate cardiac remodeling and serve as a heart failure treatment. Therefore, we identified a circRNA, named circSMAD3, that was significantly reduced in murine heart failure models. Functionally, circSMAD3 mitigated cardiomyocyte hypertrophy and inhibited cardiac fibroblast activation in vitro. Mechanistically, circSMAD3 interacts with YBX1, stabilizing it and facilitating its binding to SMAD3 in the nucleus, disrupting the TGFß/SMAD3 signaling pathway, and ultimately restoring cardiac remodeling. This study highlights circSMAD3 as a promising therapeutic target for heart failure treatment.