Dexmedetomidine enhances Mitophagy via PINK1 to alleviate hippocampal neuronal Pyroptosis and improve postoperative cognitive dysfunction in elderly rat.

Postoperative cognitive dysfunction (POCD) is a common complication in elderly surgical patients, significantly affecting their quality of life. Dexmedetomidine (Dex), an anesthetic, has shown promise in alleviating POCD, but its underlying mechanism remains unclear. This study aims to explore how Dex improves POCD in aged rats by targeting the PINK1-mediated mitochondrial autophagy pathway, reducing caspase-1/11-GSDMD-induced hippocampal neuronal pyroptosis. Transcriptome sequencing identified 300 differentially expressed genes enriched in the mitochondrial autophagy pathway in Dex-treated POCD rat hippocampal tissue, with Pink1 as a key candidate. In a POCD rat model, Dex treatment upregulated hippocampal PINK1 expression. In vitro experiments using H19-7 rat hippocampal neurons revealed that Dex enhanced mitochondrial autophagy and suppressed neuronal pyroptosis by upregulating PINK1. Further mechanistic validation demonstrated that Dex activated PINK1-mediated mitochondrial autophagy, inhibiting caspase-1/11-GSDMD-induced neuronal pyroptosis. In vivo experiments confirmed Dex's ability to reduce caspase-1/11-GSDMD-dependent hippocampal neuronal pyroptosis and improve postoperative cognitive function in aged rats. Dexmedetomidine improves postoperative cognitive dysfunction in elderly rats by enhancing mitochondrial autophagy via PINK1 upregulation, mitigating caspase-1/11-GSDMD-induced neuronal pyroptosis.

Metabolic subtypes and immune landscapes in esophageal squamous cell carcinoma: prognostic implications and potential for personalized therapies.

BACKGROUND: This study aimed to identify metabolic subtypes in ESCA, explore their relationship with immune landscapes, and establish a metabolic index for accurate prognosis assessment. METHODS: Clinical, SNP, and RNA-seq data were collected from 80 ESCA patients from the TCGA database and RNA-seq data from the GSE19417 dataset. Metabolic genes associated with overall survival (OS) and progression-free survival (PFS) were selected, and k-means clustering was performed. Immune-related pathways, immune infiltration, and response to immunotherapy were predicted using bioinformatic algorithms. Weighted gene co-expression network analysis (WGCNA) was conducted to identify metabolic genes associated with co-expression modules. Lastly, cell culture and functional analysis were performed using patient tissue samples and ESCA cell lines to verify the identified genes and their roles. RESULTS: Molecular subtypes were identified based on the expression profiles of metabolic genes, and univariate survival analysis revealed 163 metabolic genes associated with ESCA prognosis. Consensus clustering analysis classified ESCA samples into three distinct subtypes, with MC1 showing the poorest prognosis and MC3 having the best prognosis. The subtypes also exhibited significant differences in immune cell infiltration, with MC3 showing the highest scores. Additionally, the MC3 subtype demonstrated the poorest response to immunotherapy, while the MC1 subtype was the most sensitive. WGCNA analysis identified gene modules associated with the metabolic index, with SLC5A1, NT5DC4, and MTHFD2 emerging as prognostic markers. Gene and protein expression analysis validated the upregulation of MTHFD2 in ESCA. MTHFD2 promotes the progression of ESCA and may be a potential therapeutic target for ESCA. CONCLUSION: The established metabolic index and identified metabolic genes offer potential for prognostic assessment and personalized therapeutic interventions for ESCA, underscoring the importance of targeting metabolism-immune interactions in ESCA. MTHFD2 promotes the progression of ESCA and may be a potential therapeutic target for ESCA.

Structural insights into the conformational changes of BTR1/SLC4A11 in complex with PIP2.

BTR1 (SLC4A11) is a NH3 stimulated H+ (OH-) transporter belonging to the SLC4 family. Dysfunction of BTR1 leads to diseases such as congenital hereditary endothelial dystrophy (CHED) and Fuchs endothelial corneal dystrophy (FECD). However, the mechanistic basis of BTR1 activation by alkaline pH, transport activity regulation and pathogenic mutations remains elusive. Here, we present cryo-EM structures of human BTR1 in the outward-facing state in complex with its activating ligands PIP2 and the inward-facing state with the pathogenic R125H mutation. We reveal that PIP2 binds at the interface between the transmembrane domain and the N-terminal cytosolic domain of BTR1. Disruption of either the PIP2 binding site or protonation of PIP2 phosphate groups by acidic pH can transform BTR1 into an inward-facing conformation. Our results provide insights into the mechanisms of how the transport activity and conformation changes of BTR1 are regulated by PIP2 binding and interaction of TMD and NTD.

Immediate Effects of Vergence Exercises Using Automatic Dual Rotational Risley Prisms on Accommodative Lag and Facility.

INTRODUCTION: This study evaluated novel automatic dual rotational Risley prisms (ADRRPs) as a vergence exercise tool for patients with myopia to improve accommodative lag and accommodative facility. METHODS: Participants with myopia aged 20-24 years were recruited. After vergence exercises with prisms (treatment group) or plano lenses (control group) using ADRRPs for 10 min, measurements were taken using an open-field autorefractor (Grand Seiko WAM-5500) at viewing distances of 0.4 m and 6.0 m. We measured accommodative facility using a ± 2.00 D accommodative flipper. RESULTS: A total of 56 participants (treatment group, 39; control group, 17) performed vergence exercises using ADRRPs. Participants in the treatment group showed improvements in accommodative lag at a 0.4 m viewing distance, with measurements of 0.57 D (right eye; OD) and 0.53 D (left eye; OS) and 0.21 D (OD) and 0.27 D (OS) before and after the exercises, respectively (p < 0.001). Over-refractions using an open-field autorefractor with spherical equivalent contact lenses at a 6.0 m viewing distance were - 0.01 ± 0.30 D (OD) and 0.03 ± 0.34 D (OS) and 0.15 ± 0.32 D (OD) and 0.19 ± 0.28 D (OS) before and after the exercises, respectively (difference + 0.16 D; p < 0.001). Accommodative facility values before and after exercises were 14.88 ± 3.36 and 15.59 ± 3.60 cpm, respectively (p < 0.01). No significant differences in accommodative lag, relaxation, and accommodative facility before and after exercise were observed in the control group. CONCLUSIONS: Using ADRRPs in vergence exercises can improve accommodative lag, accommodative facility, and accommodative relaxation in adults with myopia. Further research to evaluate persistent and long-term effects is needed.

Structural and functional changes of the cerebellum in temporal lobe epilepsy.

Aims: This study aimed to comprehensively explore the cerebellar structural and functional changes in temporal lobe epilepsy (TLE) and its association with clinical information. Methods: The SUIT toolbox was utilized to perform cerebellar volume and diffusion analysis. In addition, we extracted the average diffusion values of cerebellar peduncle tracts to investigate microstructure alterations. Seed-based whole-brain analysis was used to investigate cerebellar-cerebral functional connectivity (FC). Subgroup analyses were performed to identify the cerebellar participation in TLE with/without hippocampal sclerosis (HS)/focal-to-bilateral tonic-clonic seizure (FBTCS) and TLE with different lateralization. Results: TLE showed widespread gray matter atrophy in bilateral crusII, VIIb, VIIIb, left crusI, and left VIIIa. Both voxel and tract analysis observed diffusion abnormalities in cerebellar afferent peduncles. Reduced FC between the right crus II and the left parahippocampal cortex was found in TLE. Additionally, TLE showed increased FCs between left lobules VI-VIII and cortical nodes of the dorsal attention and visual networks. Across all patients, decreased FC was associated with poorer cognitive function, while increased FCs appeared to reflect compensatory effects. The cerebellar structural changes were mainly observed in HS and FBTCS subgroups and were regardless of seizure lateralization, while cerebellar-cerebral FC alterations were similar in all subgroups. Conclusion: TLE exhibited microstructural changes in the cerebellum, mainly related to HS and FBTCS. In addition, altered cerebellar-cerebral functional connectivity is associated with common cognitive alterations in TLE.

The Hepatic Nerves Regulated Inflammatory Effect in the Process of Liver Injury: Is Nerve the Key Treating Target for Liver Inflammation?

Liver injury is a common pathological basis for various liver diseases. Chronic liver injury is often an important initiating factor in liver fibrosis, cirrhosis, and hepatocellular carcinoma (HCC). Currently, hepatitis A and E infections are the most common causes of acute liver injury worldwide, whereas drug toxicity (paracetamol overdose) in the USA and part of Western Europe. In recent years, chronic liver injury has become a common disease that harms human health. Meanwhile, the main causes of chronic liver injury are viral hepatitis (B, C) and long-term alcohol consumption worldwide. During the process of liver injury, massive inflammatory cytokines are stimulated by these hazardous factors, leading to a systemic inflammatory response syndrome, followed by a compensatory anti-inflammatory response, which causes immune cell dysfunction and sepsis, subsequent multi-organ failure. Cytokine release and immune cell infiltration-mediated aseptic inflammation are the most important features of the pathobiology of liver failure. From this perspective, diminishing the onset and progression of liver inflammation is of clinical importance in the treatment of liver injury. Although many studies have hinted at the critical role of nerves in regulating inflammation, there largely remains undetermined how hepatic nerves mediate immune inflammation and how the inflammatory factors released by these nerves are involved in the process of liver injury. Therefore, the purpose of this article is to summarize previous studies in the field related to hepatic nerve and inflammation as well as future perspectives on the aforementioned questions. Our findings were presented in three aspects: types of nerve distribution in the liver, how these nerves regulate immunity, and the role of liver nerves in hepatitis and liver failure.

Manganese(ii) complexes stimulate antitumor immunity via aggravating DNA damage and activating the cGAS-STING pathway.

Activating the cyclic GMP-AMP synthase-stimulator of the interferon gene (cGAS-STING) pathway is a promising immunotherapeutic strategy for cancer treatment. Manganese(ii) complexes MnPC and MnPVA (P = 1,10-phenanthroline, C = chlorine, and VA = valproic acid) were found to activate the cGAS-STING pathway. The complexes not only damaged DNA, but also inhibited histone deacetylases (HDACs) and poly adenosine diphosphate-ribose polymerase (PARP) to impede the repair of DNA damage, thereby promoting the leakage of DNA fragments into cytoplasm. The DNA fragments activated the cGAS-STING pathway, which initiated an innate immune response and a two-way communication between tumor cells and neighboring immune cells. The activated cGAS-STING further increased the production of type I interferons and secretion of pro-inflammatory cytokines (TNF-α and IL-6), boosting the tumor infiltration of dendritic cells and macrophages, as well as stimulating cytotoxic T cells to kill cancer cells in vitro and in vivo. Owing to the enhanced DNA-damaging ability, MnPC and MnPVA showed more potent immunocompetence and antitumor activity than Mn2+ ions, thus demonstrating great potential as chemoimmunotherapeutic agents for cancer treatment.

Identification of Key Aromatic Compounds in Basil (Ocimum L.) Using Sensory Evaluation, Metabolomics and Volatilomics Analysis.

Basil (Ocimum L.) is widely used as a flavor ingredient, however research on basil flavor is limited. In the current study, nine basil species were selected, including Ocimum basilicum L.var. pilosum (Willd.) Benth., Ocimum sanctum, Ocimum basilicum cinnamon, Ocimum gratissimum var. suave, Ocimum tashiroi, Ocimum basilicum, Ocimum americanum, Ocimum basilicum ct linalool, and Ocimum basilicum var. basilicum, and their fragrance and flavor characteristics were assessed by sensory evaluation. The results indicated that Ocimum basilicum var. basilicum and Ocimum gratissimum var. suave have a strong clove smell and exhibited a piquant taste. Metabolomics and volatilomics analyses measured 100 nonvolatile metabolites and 134 volatiles. Differential analysis showed that eugenol, γ-terpinene, germacrene D and malic acid were among the most varied metabolites in basil species. Combined with sensory evaluation results, correlation analysis revealed that ß-pinene and γ-cadinene contributed to the piquant smell, while eugenol and germacrene D contributed to the clove smell, and malic acid and L-(−)-arabitol contributed to the sweet flavor in basil. This study provided comprehensive flavor chemistry profiles of basil species and could be used as a guide for basil flavor improvement. The better understanding of objective sensory attributes and chemical composition of fresh basil could introduce the improved cultivars with preponderant traits, which is also in accordance with the various demands of breeders and growers, food producers, and consumers.

Comparison of methods of isolating extracellular vesicle microRNA from HepG2 cells for High-throughput sequencing.

Background: Extracellular vesicles (EVs) were reported to participate in various cellular processes based on the biomolecules, particularly microRNAs. Numerous commercial EVs isolation reagents are available. However, whether these reagents are suitable for separating EVs from the culture medium supernatant supernatant of model cell lines, such as HepG2, and whether the isolated products are suitable for High-throughput sequencing remains unclear. Methods: We examined three commonly used EVs isolation kits: the ExoQuick-TC exosome precipitation solution (EQ), Total Exosome Isolation from cell culture medium (EI), and exoEasy Maxi Kit (EM), to isolate EVs from HepG2 cell culture medium supernatants. EVs were identified based on marker proteins, particle size measurements, and electron microscopy analysis. The total amounts of microRNA and microRNA High-throughput sequencing data quality from EVs isolated by each kit were compared. Results: The total amount of EVs' microRNA isolated from the EI and EM groups were higher than that obtained from the EQ group (EQ/EI: p = 0.036, EI/EM: p = 0.024). High-throughput sequencing data quality evaluation showed that the EI group possessed higher quality than those in the EM group. Conclusion: For the cell culture medium from HepG2, EVs' microRNA isolated by EI reagents might be more suitable for High-throughput sequencing applications.

Lidocaine inhibits the proliferation and migration of endometrial cancer cells, and promotes apoptosis by inducing autophagy.

As a gynecological malignancy, endometrial cancer (EC) has a high incidence and mortality rate in women. The aim of the present study was to investigate the mechanism of EC and to identify novel effective treatment methods for this disease. The viability and proliferation of the RL95-2 human endometrial cancer cell line were assessed using Cell Counting Kit-8 assays. Colony formation, wound healing, Transwell, TUNEL and immunofluorescence assays were used to assess the effects of 5, 10 and 15 mM lidocaine on the colony formation, migration, invasiveness, apoptosis and Beclin 1 protein expression of RL95-2 cells, respectively. Furthermore, western blotting was used to analyze the protein expression levels of apoptosis- and autophagy-related proteins. The results demonstrated that lidocaine inhibited the viability, proliferation and migration of EC cells, and promoted apoptosis. Furthermore, lidocaine was demonstrated to induce autophagy and Beclin 1 protein expression in EC cells. In conclusion, lidocaine inhibited the proliferation and migration of EC cells, and promoted apoptosis via autophagy induction, which indicated that lidocaine may be a potential therapeutic drug for the treatment of EC.

Korotkoff sounds dynamically reflect changes in cardiac function based on deep learning methods.

Korotkoff sounds (K-sounds) have been around for over 100 years and are considered the gold standard for blood pressure (BP) measurement. K-sounds are also unique for the diagnosis and treatment of cardiovascular diseases; however, their efficacy is limited. The incidences of heart failure (HF) are increasing, which necessitate the development of a rapid and convenient pre-hospital screening method. In this review, we propose a deep learning (DL) method and the possibility of using K-methods to predict cardiac function changes for the detection of cardiac dysfunctions.

Crystal Structure of the Disease-Specific Protein of Rice Stripe Virus.

The rice stripe virus (RSV) is responsible for devastating effects in East Asian rice-producing areas. The disease-specific protein (SP) level in rice plants determines the severity of RSV symptoms. Isothermal titration calorimetry (ITC) and bimolecular fluorescence complementation (BiFC) assays confirmed the interaction between an R3H domain-containing host factor, OsR3H3, and RSV SP in vitro and in vivo. This study determined the crystal structure of SP at 1.71 Å. It is a monomer with a clear shallow groove to accommodate host factors. Docking OsR3H3 into the groove generates an SP/OsR3H3 complex, which provides insights into the protein-binding mechanism of SP. Furthermore, SP's protein-binding properties and model-defined recognition residues were assessed using mutagenesis, ITC, and BiFC assays. This study revealed the structure and preliminary protein interaction mechanisms of RSV SP, shedding light on the molecular mechanism underlying the development of RSV infection symptoms.

Establishment and validation of prognostic nomograms to predict the overall and cancer-specific survival in patients with hepatic malignant vascular tumors.

OBJECTIVE: To characterize the clinicopathologic features and to investigate the prognostic nomograms for overall survival (OS) and cancer-specific survival (CSS) in patients with Hepatic malignant vascular tumors (HMVT). METHOD: Patients diagnosed with HMVT between 1973 and 2015 were screened from the Surveillance, Epidemiology, and End Results (SEER) database. The Kaplan-Meier (KM) was used for survival analysis. The univariate and multivariate Cox analyses were performed to identify independent predictors. Furthermore, the prognostic nomograms were established and evaluated. RESULTS: A total of 510 HMVT patients were collected, and randomly divided into HMVT-training (N=308) and validation cohort (N=202) groups. The 3- and 5-year OS for overall HMVT were 21.3% and 19.8%, and the corresponding CSS was 29.8% and 27.7% respectively. Age at diagnosis, grade, tumor size, and histological type were identified as prognostic factors for OS and CSS in patients with HMVT. However, sex was just for predicting CSS, and T stage was only an indicator of OS. These factors were further utilized to construct the nomograms for OS and CSS in the HMVT-training cohort showing credible performance with the C-index of 0.763 and 0.762, respectively. Moreover, the AUC value for 1-, 3-, 5-year OS was 0.873, 0.905 and 0.898, and the corresponding value for CSS was 0.808, 0.794 and 0.788 respectively. Additionally, the calibration curves demonstrated a favorable agreement between the predicted and actual 1-, 3- and 5-year survival rates both in the training and validated cohorts. CONCLUSION: This was the largest population-based study to describe the clinicopathologic characteristics in patients with HMVT. Moreover, we established and validated prognostic nomograms that indicated an accurate prediction for 1-, 3- and 5-year of OS and CSS.

Structure-guided rational design of the Geobacillus thermoglucosidasius feruloyl esterase GthFAE to improve its thermostability.

Feruloyl esterases are indispensable biocatalysts catalyzing the cleavage of ester bonds between polysaccharides and their hydroxycinnamoyl cross-links. GthFAE from Geobacillus thermoglucosidasius was identified as a thermophilic alkaline feruloyl esterase with potential applications in paper manufacturing. To improve the enzymatic properties rationally and efficiently, the structure of GthFAE was solved at 1.9 Å, revealing a core domain of classical α/ß hydrolase fold and an inserted α/ß cap domain. In silico analysis based on it helped us to investigate whether the residues at the active center have positive effects on the stability, and how. Several site-directed mutations were conducted, of which substitutions at residues T41 and T150 apparently improved the thermostability. The combination mutant T41N/T150R exhibited an optimal temperature of 65 °C, a 6.4 °C higher Tm compared to wild type by 80 °C, and a 35-fold longer in half-life (201 min) at 70 °C. Molecular dynamics simulations further illustrated that the structure of T41N/T150R was more stable than the wild type and T150R stabilized the cap domain by introducing salt bridges to the region with E154 and D164. This study not only highlighted residues within the active center on their thermostability improving effects, but also contributed to the prospective industrial application of GthFAE.

Effect of nano zinc oxide on proliferation and toxicity of human gingival cells.

BACKGROUND: Periodontal dressing is used to cover the gum surface and protect the wound after periodontal surgery. Nanomaterials have been widely applied in dentistry in recent years. Zinc oxide (ZnO) is one of the main components of periodontal dressing. AIM: This study aims to explore the toxicity ZnO nanoparticles (ZnO NPs) causes to human gingival fibroblast cells (HGF-1) and its effect on cell proliferation. METHODS: First, we identified and analyzed HGF-1, including cell morphology, growth curve, and immunohistochemistry staining. Then, we treated HGF-1 with ZnO NP. Cell viability, the integrity of the cell membrane, oxidative damage, and apoptosis were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release assay, fluorescent probe, and flow cytometry. Furthermore, the expression of murine double minute 2 (MDM2) and p53 was determined by quantitative real-time polymerase chain reaction (qPCR) and Western blotting. We finally overexpressed MDM2 in HGF-1 to verify the relationship between MDM2 and cell proliferation. RESULTS: Our research indicated ZnO NPs did not affect cell proliferation at low concentrations. However, high-concentration ZnO NP inhibited cell proliferation, destroyed the integrity of cell membranes, and induced oxidative stress and apoptosis. In addition, high concentration of ZnO NPs inhibited the proliferation of HGF-1 by regulating the expression of MDM2 and p53. CONCLUSION: High concentration of ZnO NP caused toxicity to HGF-1 cells and inhibited cell proliferation by regulating MDM2 and p53 expression.

Effect of nano zinc oxide on proliferation and toxicity of human gingival cells.

BACKGROUND: Periodontal dressing is used to cover the gum surface and protect the wound after periodontal surgery. Nanomaterials have been widely applied in dentistry in recent years. Zinc oxide (ZnO) is one of the main components of periodontal dressing. AIM: This study aims to explore the toxicity ZnO nanoparticles (ZnO NPs) causes to human gingival fibroblast cells (HGF-1) and its effect on cell proliferation. METHODS: First, we identified and analyzed HGF-1, including cell morphology, growth curve, and immunohistochemistry staining. Then, we treated HGF-1 with ZnO NP. Cell viability, the integrity of the cell membrane, oxidative damage, and apoptosis were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release assay, fluorescent probe, and flow cytometry. Furthermore, the expression of murine double minute 2 (MDM2) and p53 was determined by quantitative real-time polymerase chain reaction (qPCR) and Western blotting. We finally overexpressed MDM2 in HGF-1 to verify the relationship between MDM2 and cell proliferation. RESULTS: Our research indicated ZnO NPs did not affect cell proliferation at low concentrations. However, high-concentration ZnO NP inhibited cell proliferation, destroyed the integrity of cell membranes, and induced oxidative stress and apoptosis. In addition, high concentration of ZnO NPs inhibited the proliferation of HGF-1 by regulating the expression of MDM2 and p53. CONCLUSION: High concentration of ZnO NP caused toxicity to HGF-1 cells and inhibited cell proliferation by regulating MDM2 and p53 expression.

Suboptimal Response to Tenofovir Alafenamide in Two Patients With HBeAg-Positive Hepatitis B: A Case Report.

Tenofovir alafenamide (TAF) is one of the most potent first-line nucleot(s)ide analogs for treating chronic hepatitis B virus (HBV) infections. To date, no cases of TAF drug resistance and/or suboptimal response have been reported. To our knowledge, this is the first report of two adult male patients presenting a suboptimal response response to TAF monotherapy. Our study indicates long-term observations and extensive data are needed to further evaluate the efficacy and safety of TAF, and highlights the need for the development of robust novel direct-acting antivirals and immune therapies for HBV.

Structural Insights into a Novel Esterase from the East Pacific Rise and Its Improved Thermostability by a Semirational Design.

Lipolytic enzymes are essential biocatalysts in food processing as well as pharmaceutical and pesticide industries, catalyzing the cleavage of ester bonds in a variety of acyl chain substrates. Here, we report the crystal structure of an esterase from the deep-sea hydrothermal vent of the East Pacific Rise (EprEst). The X-ray structure of EprEst in complex with the ligand, acetate, has been determined at 2.03 Å resolution. The structure reveals a unique spatial arrangement and orientation of the helix cap domain and α/ß hydrolase domain, which form a substrate pocket with preference for short-chain acyl groups. Molecular docking analysis further demonstrated that the active site pocket could accommodate p-nitrophenyl (pNP) carboxyl ligands of varying lengths (≤6 C atoms), with pNP-butyrate ester predicted to have the highest binding affinity. Additionally, the semirational design was conducted to improve the thermostability of EprEst by enzyme engineering based on the established structure and multiple sequence alignment. A mutation, K114P, introduced in the hinge region of the esterase, which displayed increased thermostability and enzyme activity. Collectively, the structural and functional data obtained herein could be used as basis for further protein engineering to ultimately expand the scope of industrial applications of marine-derived lipolytic enzymes.

The Chromosome-Level Genome of Miracle Fruit (Synsepalum dulcificum) Provides New Insights Into the Evolution and Function of Miraculin.

Miracle fruit (Synsepalum dulcificum) is a rare valuable tropical plant famous for a miraculous sweetening glycoprotein, miraculin, which can modify sour flavors to sweet flavors tasted by humans. Here, we present a chromosome-level high-quality genome of S. dulcificum with an assembly genome size of ∼550 Mb, contig N50 of ∼14.14 Mb, and 37,911 annotated protein-coding genes. Phylogenetic analysis revealed that S. dulcificum was most closely related to Camellia sinensis and Diospyros oleifera, and that S. dulcificum diverged from the Diospyros genus ∼75.8 million years ago (MYA), and that C. sinensis diverged from Synsepalum ∼63.5 MYA. Ks assessment and collinearity analysis with S. dulcificum and other species suggested that a whole-genome duplication (WGD) event occurred in S. dulcificum and that there was good collinearity between S. dulcificum and Vitis vinifera. On the other hand, transcriptome and metabolism analysis with six tissues containing three developmental stages of fleshes and seeds of miracle fruit revealed that Gene Ontology (GO) terms and metabolic pathways of "cellular response to chitin," "plant-pathogen interaction," and "plant hormone signal transduction" were significantly enriched during fruit development. Interestingly, the expression of miraculin (Chr10G0299340) progressively increased from vegetative organs to reproductive organs and reached an incredible level in mature fruit flesh, with an fragments per kilobase of transcript per million (FPKM) value of ∼113,515, which was the most highly expressed gene among all detected genes. Combining the unique signal peptide and the presence of the histidine-30 residue together composed the main potential factors impacting miraculin's unique properties in S. dulcificum. Furthermore, integrated analysis of weighted gene coexpression network analysis (WGCNA), enrichment and metabolite correlation suggested that miraculin plays potential roles in regulating plant growth, seed germination and maturation, resisting pathogen infection, and environmental pressure. In summary, valuable genomic, transcriptomic, and metabolic resources provided in this study will promote the utilization of S. dulcificum and in-depth research on species in the Sapotaceae family.

A Simple Logistic Regression Model for Predicting the Likelihood of Recurrence of Atrial Fibrillation Within 1 Year After Initial Radio-Frequency Catheter Ablation Therapy.

BACKGROUND: The clinical factors associated with the recurrence of atrial fibrillation (Af) in patients undergoing catheter ablation (CA) are still ambiguous to date. PURPOSE: 1. To recognize preoperative serologic factors and clinical features associated with Af recurrence after the first ablation treatment. 2. To Develop a Logical Regression Model for Predicting the Likelihood of Recurrence Within 1 Year After the Initial Radio-Frequency Catheter Ablation (RFCA) Therapy. METHODS: Atrial fibrillation patients undergoing RFCA at our institution from January 2016 to June 2021 were included in the analysis (n = 246). A combined dataset of relevant parameters was collected from the participants (clinical characteristics, laboratory results, and time to recurrence) (n = 200). We performed the least absolute shrinkage and selection operator (Lasso) regression with 100 cycles, selecting variables present in all 100 cycles to identify factors associated with the first recurrence of atrial fibrillation. A logistic regression model for predicting whether Af would recur within a year was created using 70% of the data as a training set and the remaining data to validate the accuracy. The predictions were assessed using calibration plots, concordance index (C-index), and decision curve analysis. RESULTS: The left atrial diameter, albumin, type of Af, whether other arrhythmias were combined, and the duration of Af attack time were associated with Af recurrence in this sample. Some clinically meaningful variables were selected and combined with recognized factors associated with recurrence to construct a logistic regression prediction model for 1-year Af recurrence. The receiver operating characteristic (ROC) curve for this model was 0.8695, and the established prediction model had a C-index of 0.83. The performance was superior to the extreme curve in the decision curve analysis. CONCLUSION: Our study demonstrates that several clinical features and serological markers can predict the recurrence of Af in patients undergoing RFCA. This simple model can play a crucial role in guiding physicians in preoperative evaluation and clinical decision-making.