Ficolin 3 promotes ferroptosis in HCC by downregulating IR/SREBP axis-mediated MUFA synthesis.

BACKGROUND: Targeting ferroptosis has been identified as a promising approach for the development of cancer therapies. Monounsaturated fatty acid (MUFA) is a type of lipid that plays a crucial role in inhibiting ferroptosis. Ficolin 3 (FCN3) is a component of the complement system, serving as a recognition molecule against pathogens in the lectin pathway. Recent studies have reported that FCN3 demonstrates inhibitory effects on the progression of certain tumors. However, whether FCN3 can modulate lipid metabolism and ferroptosis remains largely unknown. METHODS: Cell viability, BODIPY-C11 staining, and MDA assay were carried out to detect ferroptosis. Primary hepatocellular carcinoma (HCC) and xenograft models were utilized to investigate the effect of FCN3 on the development of HCC in vivo. A metabonomic analysis was conducted to assess alterations in intracellular and HCC intrahepatic lipid levels. RESULTS: Our study elucidates a substantial decrease in the expression of FCN3, a component of the complement system, leads to MUFA accumulation in human HCC specimens and thereby significantly promotes ferroptosis resistance. Overexpression of FCN3 efficiently sensitizes HCC cells to ferroptosis, resulting in the inhibition of the oncogenesis and progression of both primary HCC and subcutaneous HCC xenograft. Mechanistically, FCN3 directly binds to the insulin receptor ß (IR-ß) and its pro-form (pro-IR), inhibiting pro-IR cleavage and IR-ß phosphorylation, ultimately resulting in IR-ß inactivation. This inactivation of IR-ß suppresses the expression of sterol regulatory element binding protein-1c (SREBP1c), which subsequently suppresses the transcription of genes related to de novo lipogenesis (DNL) and lipid desaturation, and consequently downregulates intracellular MUFA levels. CONCLUSIONS: These findings uncover a novel regulatory mechanism by which FCN3 enhances the sensitivity of HCC cells to ferroptosis, indicating that targeting FCN3-induced ferroptosis is a promising strategy for HCC treatment.

Sevoflurane with Low Concentration Decrease DNA Methylation on Chronic Obstructive Pulmonary Disease (COPD)-Related Gene Promoter in COPD Rat.

Chronic obstructive pulmonary disease (COPD) is a difficult-to-cure disease that mainly affects the respiratory system. Inhaled anesthetic drug such as sevoflurane plays a controversial role in COPD by different concentration, but the underlying epigenetic mechanism remains unclear. Here, we prepared lipopolysaccharide (LPS)-induced COPD rat model, and isolated Alveolar type II (ATII) cells. We mainly focused DNA methylation on the promoter of COPD-related genes including Sftpa1, Napsa, Ca2, Sfta2, Lamp3, Wif1, Pgc, and Etv5. We observed COPD rat treated by sevoflurane with low (0.5%) and high (2%) concentrations displayed an opposite DNA methylation pattern. These six genes' promoter were all hypomethylated by 0.5% sevoflurane whereas hypermethylated by 2% sevoflurane, accompanied with the opposite transcriptional activity. We further verified that the DNMT1 binding ability contributed to DNA methylation these six genes' promoter. Moreover, we also captured DNMT1 and identified REC8 meiotic recombination protein (REC8) as the specific binding protein only existed in ATII cells treated with 0.5% sevoflurane rather than 2% and control. The binding ability of REC8 on these target genes' promoter showed highly positive correlation with DNMT1. In summary, we uncovered a potential epigenetic role of sevoflurane with low concentration in ATII cells of COPD that may help us deeply understand the pathogenesis and treatment mechanism of inhaled anesthesia drugs in COPD via a dose-dependent manner.

Characterization of a novel Tn6485h transposon carrying both blaIMP-45 and blaAFM-1 integrated into the IncP-2 plasmid in a carbapenem-resistant Pseudomonas aeruginosa.

OBJECTIVES: To characterize a carbapenem-resistant Pseudomonas aeruginosa (CRPA) with an IncP-2 plasmid containing a novel transposon, Tn6485h, which carries both blaIMP-45 and blaAFM-1. METHODS: Antimicrobial susceptibility testing and filter mating experiment were performed on PA942. The stability of the plasmid carrying both blaIMP-45 and blaAFM-1 was carried out. We determined the growth rate of the transconjugant to investigate fitness cost. Additionally, whole-genome sequencing and genomic analysis were performed on PA942. RESULTS: PA942 strain was resistant to most antibiotics except for ciprofloxacin and colistin. Bioinformatics analysis confirmed that PA942 contains an IncP-2 plasmid with a novel transposon Tn6485h carrying both blaIMP-45 and blaAFM-1. The plasmid pPA942-IMP45 can be transferred into recipient bacteria PAO1Rif with an efficiency of 2.2 × 10-7 and the transconjugant PAO1Rif/ pPA942-IMP45 can be stably inherited for 10 generations in the absence of antibiotics. CONCLUSION: We report a carbapenem-resistant P. aeruginosa strain with an IncP-2 plasmid containing a novel transposon, Tn6485h, which carries both blaIMP-45 and blaAFM-1. The IncP-2 plasmid and transposon Tn6485h may contribute to the spread of MBL genes. Therefore, effective measures to prevent the spread of these plasmids should be taken.

Three-dimensional visualization and evaluation of hilar cholangiocarcinoma resectability and proposal of a new classification.

BACKGROUND: As digital medicine has exerted profound influences upon diagnosis and treatment of hepatobiliary diseases, our study aims to investigate the accuracy of three-dimensional visualization and evaluation (3DVE) system in assessing the resectability of hilar cholangiocarcinoma (hCCA), and explores its potential clinical value. MATERIALS AND METHODS: The discovery cohort, containing 111 patients from April 2013 to December 2019, was retrospectively included to determine resectability according to revised criteria for unresectability of hCCA. 3D visualization models were reconstructed to evaluate resectability parameters including biliary infiltration, vascular involvement, hepatic atrophy and metastasis. Evaluation accuracy were compared between contrast-enhanced CT and 3DVE. Logistic analysis was performed to identify independent risk factors of R0 resection. A new comprehensive 3DVE classification of hCCA based on factors influencing resectability was proposed to investigate its role in predicting R0 resection and prognosis. The main outcomes were also analyzed in cohort validation, including 34 patients from January 2020 to August 2022. RESULTS: 3DVE showed an accuracy rate of 91% (95%CI 83.6-95.4%) in preoperatively evaluating hCCA resectability, significantly higher than 81% (95%CI 72.8-87.7%) of that of CT (p = 0.03). By multivariable analysis, hepatic artery involvement in 3DVE was identified an independent risk factor for R1 or R2 resection (OR = 3.5, 95%CI 1.4,8.8, P < 0.01). New 3DVE hCCA classification was valuable in predicting patients' R0 resection rate (p < 0.001) and prognosis (p < 0.0001). The main outcomes were internally validated. CONCLUSIONS: 3DVE exhibited a better efficacy in evaluating hCCA resectability, compared with contrast-enhanced CT. Preoperative 3DVE demonstrated hepatic artery involvement was an independent risk factor for the absence of R0 margin. 3DVE classification of hCCA was valuable in clinical practice.

Predicting the Gas Permeability of Sustainable Cement Mortar Containing Internal Cracks by Combining Physical Experiments and Hybrid Ensemble Artificial Intelligence Algorithms.

The presence of internal fissures holds immense sway over the gas permeability of sustainable cement mortar, which in turn dictates the longevity and steadfastness of associated edifices. Nevertheless, predicting the gas permeability of sustainable cement mortar that contains internal cracks poses a significant challenge due to the presence of numerous influential variables and intricate interdependent mechanisms. To solve the deficiency, this research establishes an innovative machine learning algorithm via the integration of the Mind Evolutionary Algorithm (MEA) with the Adaptive Boosting Algorithm-Back Propagation Artificial Neural Network (ABA-BPANN) ensemble algorithm to predict the gas permeability of sustainable cement mortar that contains internal cracks, based on the results of 1452 gas permeability tests. Firstly, the present study employs the MEA-tuned ABA-BPANN model as the primary tool for gas permeability prediction in cement mortar, a comparative analysis is conducted with conventional machine learning models such as Particle Swarm Optimisation Algorithm (PSO) and Genetic Algorithm (GA) optimised ABA-BPANN, MEA optimised Extreme Learning Machine (ELM), and BPANN. The efficacy of the MEA-tuned ABA-BPANN model is verified, thereby demonstrating its proficiency. In addition, the sensitivity analysis conducted with the aid of the innovative model has revealed that the gas permeability of durable cement mortar incorporating internal cracks is more profoundly affected by the dimensions and quantities of such cracks than by the stress conditions to which the mortar is subjected. Thirdly, puts forth a novel machine-learning model, which enables the establishment of an analytical formula for the precise prediction of gas permeability. This formula can be employed by individuals who lack familiarity with machine learning skills. The proposed model, namely the MEA-optimised ABA-BPANN algorithm, exhibits significant potential in accurately estimating the gas permeability of sustainable cement mortar that contains internal cracks in varying stress environments. The study highlights the algorithm's ability to offer essential insights for designing related structures.

The role of main pancreatic duct stent in the enucleation of benign/borderline pancreatic head tumors: a cohort study.

PURPOSE: This study aimed to reveal the role of preoperative main pancreatic duct (MPD) stent placement in reducing the intraoperative main pancreatic duct injury rate and the incidence of postoperative pancreatic leakage following pancreatic tumor enucleation. METHODS: A retrospective cohort analysis was performed for all patients with benign/borderline pancreatic head tumors who were treated with enucleation. The patients were divided into two groups (standard vs. stent) depending on whether they underwent main pancreatic duct stent placement prior to surgery. RESULTS: Thirty-three patients were finally included in the analytical cohort. Compared with the standard group, patients in the stent group had a shorter distance between tumors and main pancreatic duct (p=0.01) and presented with larger tumors (p<0.01). The rates of POPF (grade B&C) were 39.1% (9/23) and 20% (2/10) in the standard and stent groups, respectively (p<0.01). Major postoperative complications occurred more frequently in the standard group than in the stent group (14 versus 2; p<0.01). No significant differences in mortality, in-hospital stay or medical cost were observed between the two groups (p>0.05). CONCLUSIONS: MPD stent placement prior to surgery may facilitate pancreatic tumor enucleation, minimize MPD injury and decrease the occurrence of postoperative fistula.

hENT1's role in adjuvant intra-arterial gemcitabine-based chemotherapy for resectable pancreatic cancer patients.

BACKGROUND: We aimed to verify the role of hENT1 as a prognostic predictor for patients with resectable pancreatic ductal adenocarcinoma (PDAC) who underwent radical resection followed by intra-arterial infusion of gemcitabine-based regimen. METHODS: We collected surgical samples from 102 patients with resectable PDAC who received radical resection followed by intra-arterial infusion of gemcitabine-based regimen. The hENT1 expression with the help of immunohistochemistry was conducted using formalin-fixed and paraffin embedded tissues. The Kaplan-Meier analyses and Cox regression were used to evaluate the mortality hazard associated with the discrepancy between strong and weak of hENT1 expression. Patients' clinical and pathological characteristics were compared between the two groups, then the role of hENT1 as a prognostic predictor was further explored. RESULTS: A total of 102 patients were included to assess the hENT1 expression. 50 patients were classified into high hENT1 expression group, the other 52 patients were attributed into low hENT1 expression group. High hENT1 expression was related to a significantly improved overall survival (OS) (p = 0.014) and disease-free survival (DFS) (p = 0.004). Both univariate (p = 0.001) and multivariate analyses (p < 0.001) indicated that high hENT1 expression was related to a decreased mortality. CONCLUSIONS: High expression of hENT1 is positive prognostic factor for adjuvant intra-arterial gemcitabine-based chemotherapy in resectable PDAC.

The Effect of the Histone Chaperones HSPA8 and DEK on Tumor Immunity in Hepatocellular Carcinoma.

Complex immune contexture leads to resistance to immunotherapy in hepatocellular carcinoma (HCC), and the need for new potential biomarkers of immunotherapy in HCC is urgent. Histone chaperones are vital determinants of gene expression and genome stability that regulate tumor development. This study aimed to investigate the effect of histone chaperones on tumor immunity in HCC. Bioinformatics analyses were initially performed using The Cancer Genome Atlas (TCGA) database, and were validated using the Gene Expression Omnibus (GEO) database and the International Cancer Genome Consortium (ICGC) database. Immune-related histone chaperones were screened with the Spearman rank coefficient. Consensus clustering was utilized to divide the HCC samples into two clusters. ESTIMATE, CIBERSORT and ssGSEA analyses were performed to assess immune infiltration. The expression of immunomodulatory genes, chemokines and chemokine receptors was analyzed to evaluate sensitivity to immunotherapy. The differentially expressed genes (DEGs) were included in weighted gene coexpression network analysis (WGCNA) to identify the hub genes. Enrichment analyses were used to investigate the functions of the hub genes. The Kaplan-Meier method and log-rank test were conducted to draw survival curves. A Cox regression analysis was utilized to identify independent risk factors affecting prognosis. HSPA8 and DEK were screened out from 36 known histone chaperones based on their strongest correlation with the ESTIMATE score. Cluster 2, with high HSPA8 expression and low DEK expression, tended to have stronger immune infiltration and better sensitivity to immunotherapy than Cluster 1, with low HSPA8 expression and high DEK expression. Furthermore, WGCNA identified 12 hub genes closely correlated with immune infiltration from the DEGs of the two clusters, of which FBLN2 was proven to be an independent protective factor of HCC patients. HSPA8 and DEK are expected to be biomarkers for precisely predicting the effect of immunotherapy, and FBLN2 is expected to be a therapeutic target of HCC.

Hepatolithiasis Classification Based on Anatomical Hepatectomy.

BACKGROUND: This study aims to propose a novel classification system to standardize the treatment of hepatolithiasis. METHODS: A hepatolithiasis classification named LHO was proposed to represent the distribution of stones in the segmental bile ducts and the hepatic atrophy associated with the stones (L), the existence of stones or strictures in the hilar bile duct (H), and dysfunction of the Oddi sphincter (O), which can be used to formulate ideal surgical protocols. One hundred and forty-seven primary hepatolithiasis patients treated between 2013 and 2018 were classified into different types and divided into two groups. If the patient's actual surgical procedure matched the ideal surgical protocol, the patients were included in the matching group; otherwise, patients were included in the nonmatching group. The rates of residual stones, recurrence, and a good quality of life (QOL) were analyzed among the patients in the matching and nonmatching groups and previous reports. RESULTS: According to the classification of each patient, 77.6% of the patients were included in the matching group, and 22.4% were included in the nonmatching group. The rates of residual stones, recurrence, and a good QOL were significantly better in the matching group than in the nonmatching group (9.6% vs. 27.3%; 8.0% vs. 35.0%; 89.5% vs. 65.4%); the rates of residual stones and a good QOL were also better than those in previous reports (9.6% vs. 19.1%; 89.5% vs. 61.6%). CONCLUSIONS: The LHO classification can comprehensively reflect the key points of treatment, which is beneficial for formulating effective and standardized surgical plans of hepatolithiasis.

MicroRNA-216a targets WT1 expression and regulates KRT7 transcription to mediate the progression of pancreatic cancer-A transcriptome analysis.

Gene expression profiling has been broadly performed in the field of cancer research. This study aims to explore the key gene regulatory network and focuses on the functions of microRNA (miR)-216a in pancreatic cancer (PC). PC datasets GSE15471, GSE16515, and GSE32676 were used to screen the differentially expressed genes (DEGs) in PC. A miRNA microarray analysis and gene oncology analysis suggested miR-216a as an important differentially expressed miRNA in PC. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis suggested that miR-216a and the DEGs are largely enriched on the phosphatidyl inositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway. miR-216a targeted Wilms Tumor 1 (WT1), while WT1 promoted transcription activity of keratin 7 (KRT7). Upregulation of miR-216a reduced proliferation and invasiveness of PC cells, while further upregulation of WT1 blocked the functions of miR-216a. Silencing of KRT7 diminished the oncogenic role of WT1. The in vitro results were reproduced in vivo. High expression of miR-216a while poor expression of WT1 indicated better prognosis of PC patients. The miR-216a/WT1/KRT7 axis influenced the activity of the PI3K/AKT pathway. To conclude, this study evidenced that miR-216a suppressed WT1 expression and blocked KRT7 transcription, which inactivated the PI3K/AKT signaling and reduced PC progression.

A high-quality apple genome assembly reveals the association of a retrotransposon and red fruit colour.

A complete and accurate genome sequence provides a fundamental tool for functional genomics and DNA-informed breeding. Here, we assemble a high-quality genome (contig N50 of 6.99 Mb) of the apple anther-derived homozygous line HFTH1, including 22 telomere sequences, using a combination of PacBio single-molecule real-time (SMRT) sequencing, chromosome conformation capture (Hi-C) sequencing, and optical mapping. In comparison to the Golden Delicious reference genome, we identify 18,047 deletions, 12,101 insertions and 14 large inversions. We reveal that these extensive genomic variations are largely attributable to activity of transposable elements. Interestingly, we find that a long terminal repeat (LTR) retrotransposon insertion upstream of MdMYB1, a core transcriptional activator of anthocyanin biosynthesis, is associated with red-skinned phenotype. This finding provides insights into the molecular mechanisms underlying red fruit coloration, and highlights the utility of this high-quality genome assembly in deciphering agriculturally important trait in apple.

Preoperative Low-dose and High-dose Pregabalin and Cardiovascular Response to Endotracheal Intubation: A Prospective, Randomized, Single-blind, Controlled Study in China.

PURPOSE: A prospective, randomized, single-blind, controlled clinical study was designed to evaluate the efficacy and tolerability of preoperative pregabalin on cardiovascular response to laryngoscopy and endotracheal intubation. METHODS: Patients aged 18-60 years with an American Society of Anesthesiologists scale score of I or II were recruited and randomly allocated to receive placebo (control), low-dose (150-mg) pregabalin, or high-dose (300-mg) pregabalin. The medications were orally administered 1 hour before general anesthesia. Heart rate, systolic and diastolic blood pressures, and mean arterial blood pressure were measured and recorded prior to the administration of placebo or pregabalin; before endotracheal intubation; and at 0, 1, 3, 5, 7, and 10 minutes after intubation. The sedation score was evaluated 1 hour after the administration of placebo or pregabalin. FINDINGS: A total of 90 patients were enrolled (n = 30 per group). Pregabalin (150 or 300 mg) was associated with reduced blood pressure fluctuations after intubation, but with no significant differences between the 2 dose groups. Pregabalin was associated with an inhibitory effect on heart rate fluctuations and reduced hemodynamic complications after intubation, in a dose-dependent manner, but no effect on the required perioperative opioid dosage was found. Both doses were effective in reducing preoperative anxiety, but visual analog scale pain scores at 1 hour after surgery were reduced only in limb and spine as well as abdominal surgeries. A pregabalin-related adverse reaction was dizziness, which was observed at 1 hour after surgery in both groups. IMPLICATIONS: In this study, high-dose (300-mg) pregabalin effectively attenuated cardiovascular response after endotracheal intubation. ClinicalTrials.gov identifier: NCT03456947.