Search

Abstract Cardiac regenerative potential in the animal kingdom displays striking divergence across ontogeny and phylogeny. Here we discuss several fundamental questions in heart regeneration and provide both a holistic view of heart regeneration in the organism as a whole, as well as a single-cell perspective on intercellular communication among diverse cardiac cell populations. We hope to provide valuable insights that advance our understanding of organ regeneration and future therapeutic strategies.

Abstract Propionic acidemia (PA), resulting from Pcca or Pccb gene mutations, impairs propionyl-CoA metabolism and induces metabolic alterations. While speculation exists that fasting might exacerbate metabolic crises in PA patients by accelerating the breakdown of odd-chain fatty acids and amino acids into propionyl-CoA, direct evidence is lacking. Our investigation into the metabolic effects of fasting in Pcca -/-(A138T) mice, a PA model, reveals surprising outcomes. Propionylcarnitine, a PA biomarker, decreases during fasting, along with the C3/C2 (propionylcarnitine/acetylcarnitine) ratio, ammonia, and methylcitrate. Although moderate amino acid catabolism to propionyl-CoA occurs with a 23-h fasting, a significant reduction in microbiome-produced propionate and increased fatty acid oxidation mitigate metabolic alterations by decreasing propionyl-CoA synthesis and enhancing acetyl-CoA synthesis. Fasting-induced gluconeogenesis further facilitates propionyl-CoA catabolism without changing propionyl-CoA carboxylase activity. These findings suggest that fasting may alleviate metabolic alterations in Pcca -/-(A138T) mice, prompting the need for clinical evaluation of its potential impact on PA patients.

A number of CaHfO3 modified BiFeO3-0.33BaTiO3 (BF-0.33BT-xCH) lead-free piezoceramics were fabricated through the solid-state sintering method and comprehensively investigated in this work. Under the optimal sintering temperature, all compositions display a typical perovskite structure in a pseudo-cubic phase with slightly larger lattice parameters as the CH content increases. The electrical resistivity is highly enhanced due to the addition of CH. Microstructures, including the grain morphology, core-shell structure, and chemistry inhomogeneities, are demonstrated upon different BF-0.33BT-xCH compositions. In particular, the core-shell structures with non-uniform element distributions in the compositions can be eliminated by adding sufficient CH content (x > 0.05). The highest saturation polarization (40.1 μC/cm2), remnant polarization (26.8 μC/cm2), and converse piezoelectric coefficient (290 pm/V) are achieved in the BF-0.33BT-0.01CH piezoceramic, which are significantly enhanced in comparison with the undoped BF-0.33BT piezoceramic. With further increasing the CH content, the piezoelectric properties of BF-0.33BT-xCH ceramics decline rapidly, and they start to exhibit characteristics of relaxor ferroelectrics.

IntroductionLeritrelvir (RAY1216) acts as a main protease inhibitor that hinders the cleavage of viral precursor proteins, thereby inhibiting virus replication of SARS-CoV-2). This antiviral mechanism has shown significant efficacy against the novel coronavirus. Preclinical studies have demonstrated the potent antiviral activity and favorable safety profile of this compound. This study aims to develop a pharmacokinetic model for leritrelvir, with and without ritonavir as a pharmacokinetic enhancer and to evaluate the necessity of co-administration with ritonavir and to investigate different dosage regimens.MethodThe model establishment was based on plasma concentration data from a phase I trial involving 72 subjects in single-ascending dose (SAD), multiple-ascending dose (MAD), and a food effect cohort. Analysis was conducted using a nonlinear mixed-effects model, and clinical trial simulations were carried out.ResultsThe findings of this study demonstrate a favorable safety profile for leritrelvir. With simulation suggests that a 400 mg thrice-daily (TID) regimen may be optimal to maintain the trough concentrations (Ctrough) above levels required for inhibiiting viral replication. While ritonavir was found to enhance exposure, it was deemed unnecessary. Gender and food consumption were identified as significant covariates affecting pharmacokinetic parameters, however, no dose adjustments were deemed necessary.DiscussionThis findings supported by subsequent phase II and phase III trials validated the appropriateness of a 400 mg TID regimen for the administration of leritrelvir.

Summary: Liquid-liquid phase separation (LLPS) mediated by G3BP1/2 proteins and non-translating mRNAs mediates stress granule (SG) assembly. We investigated the phylogenetic evolution of G3BP orthologs from unicellular yeast to mammals and identified both conserved and divergent features. The modular domain organization of G3BP orthologs is generally conserved. However, invertebrate orthologs displayed reduced capacity for SG assembly in human cells compared to vertebrate orthologs. We demonstrated that the protein-interaction network facilitated by the NTF2L domain is a crucial determinant of this specificity. The evolution of the G3BP1 network coincided with its exploitation by certain viruses, as evident from the interaction between viral proteins and G3BP orthologs in insects and vertebrates. We revealed the importance and divergence of the G3BP interaction network in human SG formation. Leveraging this network, we established a 7-component in vitro SG reconstitution system for quantitative studies. These findings highlight the significance of G3BP network divergence in the evolution of biological processes.

IntroductionEndometriosis (EMs) is characterized by ectopic growth of active endometrial tissue outside the uterus. The Luoshi Neiyi prescription (LSNYP) has been extensively used for treating EMs in China. However, data on the active chemical components of LSNYP are insufficient, and its pharmacological mechanism in EMs treatment remains unclear. This study aimed to explore the potential mechanism of LSNYP for EMs through network pharmacology based on the components absorbed into the blood.MethodsUltra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry was used to analyze blood components, and a series of network pharmacology strategies were utilized to predict targets of these components and EMs. Protein–protein interaction (PPI) network analysis, component–target–disease network construction, gene ontology (GO) functional enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. Additionally, molecular docking, molecular dynamics simulations, and in vitro and in vivo experiments were conducted to validate the HIF1A/EZH2/ANTXR2 pathway associated with hypoxic pathology in EMs.ResultsThirty-four absorbed components suitable for network pharmacology analysis were identified, and core targets, such as interleukin 6, EGFR, HIF1A, and EZH2, were founded. Enrichment results indicated that treatment of EMs with LSNYP may involve the regulation of hypoxia and inflammatory-related signaling pathways and response to oxidative stress and transcription factor activity. Experimental results demonstrated that LSNYP could decrease the expression of HIF1A, ANTXR2, YAP1, CD44, and β-catenin, and increased EZH2 expression in ectopic endometrial stromal cells and endometriotic tissues. Molecular docking and molecular dynamics simulations manifested that there was stable combinatorial activity between core components and key targets of the HIF1A/EZH2/ANTXR2 pathway.ConclusionLSNYP may exert pharmacological effects on EMs via the HIF1A/EZH2/ANTXR2 pathway; hence, it is a natural herb-related therapy for EMs.

To address the challenges of low detection rate and high missed detection rate of military aircraft in current complex remote sensing data, and to meet the requirements of real-time detection and easy deployment of models, this article introduces DET-you only look once (YOLO), an innovative detection model. First, to tackle the issue of reduced accuracy in identifying small targets amidst intricate backgrounds, a novel feature extraction component, C2f_DEF, was devised. This module replaced all existing C2f components within YOLOv8n, thereby significantly enhancing the model's ability to cope with complicated environmental contexts. Second, to achieve the functionality of easy deployment of the model, some deep structures were simplified to make the model more lightweight. Afterward, to further improve the model's ability to handle complex backgrounds and dense environments in remote sensing images and to improve the model's detection accuracy for military aircraft, the DAT module was embedded in the model. Finally, this article also optimized the loss function and reg_max to further reduce computational costs while improving the detection accuracy of the model. To verify the effectiveness and strong universality of DET-YOLO, extensive experimental verification was conducted on three publicly available datasets, namely MAR20, NWPU VHR-10, and NEU-DET. On the MAR20 dataset, compared with other advanced models, DET-YOLO achieved the highest mAP0.5 (namely 94.7%) with only 80 training epochs while meeting lightweight and real-time requirements. While on the other two datasets, DET-YOLO also achieved the best detection performance.

ABSTRACT: Inhibiting oxidative stress is key for ensuring sperm motility during semen cryopreservation. The aim of this study was to investigate the effect of adding alpha-lipoic acid (ALA) as an extender in rooster semen cryopreservation. Different concentrations of ALA were added to the frozen diluent of rooster semen; subsequently, computer-aided semen analysis was used to determine membrane functional integrity, acrosome integrity, antioxidant capacity (based on T-AOC, GSH-Px, SOD, CAT, and MDA contents), and mitochondrial integrity. The frozen sperm ultrastructure was observed using transmission electron microscopy. The results showed that the addition of different concentrations of ALA partially to greatly improved the quality of frozen sperm; in particular, 8 μg/mL ALA significantly improved multiple parameters of sperm quality, including sperm motility and antioxidant enzyme activity, after freeze–thaw. The results of this study provide empirical and theoretical support for effective rooster semen cryopreservation and can inform the development of new protective agents in the field of livestock reproduction.

Abstract Non-alcoholic fatty liver disease is a growing health burden with limited treatment options worldwide. Herein we report a randomized, double-blind, placebo-controlled, multiple-dose trial of a first-in-class pan-phosphodiesterase inhibitor ZSP1601 in 36 NAFLD patients (NCT04140123). There were three cohorts. Each cohort included twelve patients, nine of whom received ZSP1601 50 mg once daily, 50 mg twice daily, or 100 mg twice daily, and three of whom received matching placebos for 28 days. The primary outcomes were the safety and tolerability of ZSP1601. A total of 27 (27/36, 75%) patients experienced at least one treatment-emergent adverse event (TEAE). Most TEAEs were mild to moderate. There was no Serious Adverse Event. Diarrhea, transiently elevated creatinine and adaptive headache were frequently reported adverse drug reaction. We conclude that ZSP1601 is well-tolerated and safe, showing effective improvement in liver chemistries, liver fat content and fibrosis in patients with NAFLD.

Abstract Propionic acidemia (PA) is a rare autosomal recessive congenital disease caused by mutations in the PCCA or PCCB genes. Elevated propionylcarnitine, 2-methylcitric acid (2MCA), propionylglycine, glycine and 3-hydroxypropionate can be used to diagnose PA. Early-onset PA can lead to acute deterioration, metabolic acidosis, and hyperammonemia shortly after birth, which can result in high mortality and disability. Late-onset cases of PA have a more heterogeneous clinical spectra, including growth retardation, intellectual disability, seizures, basal ganglia lesions, pancreatitis, cardiomyopathy, arrhythmias, adaptive immune defects, rhabdomyolysis, optic atrophy, hearing loss, premature ovarian failure, and chronic kidney disease. Timely and accurate diagnosis and appropriate treatment are crucial to saving patients’ lives and improving their prognosis. Recently, the number of reported PA cases in China has increased due to advanced diagnostic techniques and increased research attention. However, an overview of PA prevalence in China is lacking. Therefore, this review provides an overview of recent advances in the pathogenesis, diagnostic strategies, and treatment of PA, including epidemiological data on PA in China. The most frequent variants among Chinese PA patients are c.2002G > A in PCCA and c.1301C > T in PCCB, which are often associated with severe clinical symptoms. At present, liver transplantation from a living (heterozygous parental) donor is a better option for treating PA in China, especially for those exhibiting a severe metabolic phenotype and/or end-organ dysfunction. However, a comprehensive risk–benefit analysis should be conducted as an integral part of the decision-making process. This review will provide valuable information for the medical care of Chinese patients with PA.

Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) is one of the most devastating agricultural pests worldwide due to its high reproductive and invasive abilities. The elucidation of its gonadal developmental characteristics and the identification of sex-related genes will provide a useful genetic basis for reproductive-based pest control. Here, the gonadal transcriptome of B. dorsalis was sequenced, and novel gonad-specific expressed genes were analyzed. A total of 1338, 336, 35, and 479 differentially expressed genes (DEGs) were found in the testis (TE), ovary (OV), female accessory gland (FAG), and male accessory gland (MAG), respectively. Furthermore, 463 highly expressed gonad-specific genes were identified, with the TE having the highest number of specific highly expressed genes, at 402, followed by 51 in the OV, 9 in the MAG, and only 1 in the FAG. Strikingly, approximately half of highly expressed gonad-specific genes were uncharacterized. Then, it was found that 35, 17, 3, 2, and 1 of 202 uncharacterized highly expressed TE-specific genes encoded proteins that contained transmembrane domains, signal peptides, high-mobility group boxes, the zinc finger domain, and the BTB/POZ domain, respectively. Interestingly, approximately 40% of uncharacterized highly expressed gonad-specific genes encoding proteins were not predicted to possess functional motifs or domains. Finally, the spatiotemporal expression and sequence characterization of six novel highly expressed gonad-specific genes were analyzed. Altogether, our findings provide a valuable dataset for future functional analyses of sex-related genes and potential target sites for pest control.

The air fryer utilizes heated air rather than hot oil to achieve frying, eliminating the need for cooking oil, rendering it a healthier cooking method than traditional frying and baking. However, there is limited evidence supporting that the air fryer could effectively reduce the level of food-derived carcinogen. In this study, we compared the concentration of Benzo[a]pyrene (BaP), a typical carcinogen, in beef patties cooked using an air fryer and an oven, under different cooking conditions, including temperatures (140 °C, 160 °C, 180 °C, and 200 °C), times (9, 14, and 19 min), and oil added or not. The adjusted linear regression analysis revealed that the BaP concentration in beef cooked in the air fryer was 22.667 (95% CI: 15.984, 29.349) ng/kg lower than that in beef cooked in the oven. Regarding the air fryer, the BaP concentration in beef cooked without oil brushing was below the detection limit, and it was significantly lower than in beef cooked with oil brushing (p < 0.001). Therefore, cooking beef in the air fryer can effectively reduce BaP concentration, particularly due to the advantage of oil-free cooking, suggesting that the air fryer represents a superior option for individuals preparing meat at high temperatures.

BiFeO3–BaTiO3 is a promising lead-free piezoelectric ceramic, exhibiting high Curie temperature and superior electrochemical characteristics. In this work, [Formula: see text]BiFeO3–xBaTiO3 (BF–xBT, [Formula: see text], 0.28, 0.30, 0.32, 0.34, 0.36) ceramics were fabricated using the conventional solid-state reaction method through precise composition control. Multiple characterization techniques, including X-ray powder diffraction (XRD), scanning electron microscope (SEM), and electrical property testing systems, were applied to systematically examine the crystallographic structure, microstructure, as well as the dielectric, ferroelectric and piezoelectric properties of the BF–xBT ceramics. The XRD results confirm that all compositions exhibit a typical perovskite structure, transitioning from a single rhombohedral phase to a rhombohedral–cubic phase mixture as the BT content increases. SEM shows apparent core–shell microstructures in the ceramics. Notably, the results demonstrated that the BF–0.30BT ceramic exhibits the maximum piezoelectric constant ([Formula: see text]) [Formula: see text][Formula: see text]pC/N, while the BF–0.34BT ceramic displays the maximum converse piezoelectric constant [Formula: see text][Formula: see text]pm/V, which highlights the suitability of BF–BT ceramics for high-performance piezoelectric applications.

Abstract Transcranial direct current stimulation (tDCS) of the occipital lobe may modulate visual cortex neuroplasticity. We assessed the acute effect of visual cortex anodal (a-)tDCS on ocular dominance plasticity induced by short-term monocular deprivation (MD), a well-established technique for inducing homeostatic plasticity in the visual system. In Experiment 1, active or sham visual cortex tDCS was applied during the last 20 min of 2-h MD following a within-subjects design (n = 17). Ocular dominance was measured using two computerized tests. The magnitude of ocular dominance plasticity was unaffected by a-tDCS. In Experiment 2 (n = 9), we investigated whether a ceiling effect of MD was masking the effect of active tDCS. We replicated Experiment 1 but used only 30 min of MD. The magnitude of ocular dominance plasticity was decreased with the shorter intervention, but there was still no effect of active a-tDCS. Within the constraints of our experimental design and a-tDCS parameters, visual cortex a-tDCS did not modulate the homeostatic mechanisms that drive ocular dominance plasticity in participants with normal binocular vision.

Since the commencement of Sustainable Development Goals (SDGs), renewable energy has faced many challenges in reaching the target of SDGs, while the potential ecological impact on the environment cannot be ignored. The expansion of photovoltaic (PV) networks is raising concerns regarding the potential impact of large-scale PV power stations on local ecosystems. However, a comprehensive understanding of the specific responses of vegetation and soil factors to PV con-struction across different study locations is still lacking. To address this knowledge gap, we con-ducted a comprehensive meta-analysis of 28 studies internationally representing 31 observational points that evaluated 432 different vegetation and soil factor responses to the installation of PV power stations. We used piecewiseSEM to explore the responses of predictors/factors to the eco-logical environment. This study investigated the geographical and environmental conditions associated with PV construction and their responses to vegetation and soil factors, considering the advantages and disadvantages of PV power station construction in different ecosystems. The results indicate that (1) the response of the ecosystems to PV power station construction increased by 58.89%. Among these, the most significant improvement is in the desert, which accounts for 77.26%. Im-provement in temperate regions is 59.62%, while there is a decrease of 19.78% in boreal regions. Improvement in arid regions is 84.45%, while improvement in humid regions is 9.84%. (2) PV construction promotes SWC, vegetation diversity, vegetation coverage, and vegetation biomass, significantly enhancing vegetation productivity. (3) Among the different ecosystems, PV power station effects were most significant in deserts, while showing negative impacts on croplands. (4) Compared to below-panel treatments, between-panel treatments were more effective in improving ecological conditions. The study contributes to mitigating adverse effects associated with photovoltaic site development, offering insights into site selection planning for solar power stations and the advancement of the renewable energy sector.

Objective: NRF2 is a master transcription factor that regulates the stress response. NRF2 is frequently mutated and activated in human esophageal squamous cell carcinoma (ESCC), which drives resistance to chemotherapy and radiation therapy. Therefore, a great need exists for NRF2 inhibitors for targeted therapy of NRF2high ESCC. Design: We performed high-throughput screening of two compound libraries from which hit compounds were further validated in human ESCC cells and a genetically modified mouse model. The mechanism of action of one compound was explored by biochemical assays. Results: Using high-throughput screening of two small molecule compound libraries, we identified 11 hit compounds as potential NRF2 inhibitors with minimal cytotoxicity at specified concentrations. We then validated two of these compounds, pyrimethamine and mitoxantrone, by demonstrating their dose- and time-dependent inhibitory effects on the expression of NRF2 and its target genes in two NRF2Mut human ESCC cells (KYSE70 and KYSE180). RNAseq and qPCR confirmed the suppression of global NRF2 signaling by these two compounds. Mechanistically, pyrimethamine reduced NRF2 half-life by promoting NRF2 ubiquitination and degradation in KYSE70 and KYSE180 cells. Expression of an Nrf2E79Q allele in mouse esophageal epithelium (Sox2CreER;LSL-Nrf2E79Q/+) resulted in an NRF2high phenotype, which included squamous hyperplasia, hyperkeratinization, and hyperactive glycolysis. Treatment with pyrimethamine (30 mg/kg/day, p.o.) suppressed the NRF2high esophageal phenotype with no observed toxicity. Conclusion: We have identified and validated pyrimethamine as an NRF2 inhibitor that may be rapidly tested in the clinic for NRF2high ESCC.

Abstract Baiyang Lake is the largest freshwater lake in North China, playing an important role in aquatic products production and eco-environment improvement. Traditional organochlorine pesticides were not enough to reflect ecological risk. We performed the high-throughput and non-targeted screening to identify the high-residue and wide-distribution pesticides at farmland-river interlaced area. We firstly reported the residue level and spatio-temporal distribution of typical pesticides in soils and waters (SP1–SP13) near Fuhe river in 2020–2021. The mean recoveries of eight pesticides ranged from 79.4 to 129%. The residues were 0.250–3530 ng/L (water) and 2.79 × 10−3–647 μg/kg dw (soil), respectively. Thiamethoxam was dominant with the high-residue proportion (HRP) of 53–95% (water, HRP > 50%) and 63–97% (soil, HRP > 60%), respectively. Most of pesticides almost have no significant season-change. The risk quotient (RQ) model results showed that although most pesticides have no aquatic risk (RQ

Abstract Limited efforts have been made in assessing the effect of genome-wide profiling of RNA splicing-related variation on lung cancer risk. In the present study, we first identified RNA splicing-related genetic variants linked to lung cancer in a genome-wide profiling analysis and then conducted a two-stage (discovery and replication) association study in populations of European ancestry. Discovery and validation were conducted sequentially with a total of 29,266 cases and 56,450 controls from both the Transdisciplinary Research in Cancer of the Lung and the International Lung Cancer Consortium as well as the OncoArray database. For those variants identified as significant in the two datasets, we further performed stratified analyses by smoking status and histological type and investigated their effects on gene expression and potential regulatory mechanisms. We identified three genetic variants significantly associated with lung cancer risk: rs329118 in JADE2 (P = 8.80E−09), rs2285521 in GGA2 (P = 4.43E−08), and rs198459 in MYRF (P = 1.60E−06). The combined effects of all three SNPs were more evident in lung squamous cell carcinomas (P = 1.81E−08, P = 6.21E−08, and P = 7.93E−04, respectively) than in lung adenocarcinomas and in ever smokers (P = 9.80E−05, P = 2.70E−04, and P = 2.90E−05, respectively) than in never smokers. Gene expression quantitative trait analysis suggested a role for the SNPs in regulating transcriptional expression of the corresponding target genes. In conclusion, we report that three RNA splicing-related genetic variants contribute to lung cancer susceptibility in European populations. However, additional validation is needed, and specific splicing mechanisms of the target genes underlying the observed associations also warrants further exploration.

With the development of high-density and highly integrated electronic devices, the insulation characteristics between the interconnects of printed circuit boards (PCBs) are the key issues to be considered in their design and production. Most of the existing studies on insulation characteristics are based on parallel interconnect structures; however, the possibility of flashover or breakdown on PCB at parallel interconnects is relatively low compared to other irregular structures. The flashover characteristics of PCB interconnects need to be more comprehensively presented. In this paper, the commonly used interconnects were divided into three typical structures. Under the same DC excitation, the “interconnect with pad” structure had the most severe electric field concentration, and the insulation characteristic of this structure was the worse of the three. Furthermore, the effects of the distance between the interconnect and pad size on the flashover voltages of the “interconnect with pad” structure were investigated and explained by the maximum electric field strength, which could be approximately calculated by the existing equation.

Land degradation destroys human habitats, and vegetation is a marker reflecting land degradation. In this article, the Balochistan Province of Pakistan, which has a fragile ecological environment, was selected as a typical case to analyze its land degradation over 21 years. Relevant studies that used the NDVI and remote sensing data to monitor land degradation already existed. Based on the data product of MODIS, this study obtained the spatio-temporal trends of the normalized difference vegetation index (NDVI) changes from 2000 to 2020 using the sen+ Mann–Kendall (MK) test and Hurst index and analyzed the driving factors of land degradation and restoration by employing the multiple stepwise regression method. The residual analysis method was an effective tool for distinguishing between anthropogenic and climatic impacts, given that not all regions have a significant correlation between the NDVI and rainfall. The main climatic drivers of the NDVI were derived based on the Geodetector analysis and stripped of the main climatic factors by residual analysis to explore the influence of anthropogenic factors on the NDVI. The results show the following: (1) Balochistan is dominated by land restoration. Land restoration is mainly dominated by climate as well as both climate and human factors, and land degradation is mainly dominated by climate and human factors. (2) The Geodetector-based study found high correlations between the NDVI and TMP, MAP, AET and PET, complementing most previous residual analyses that considered only precipitation and temperature. In Balochistan, TMP, AET, PET and MAP were the dominant climatic factors affecting the spatial distribution of the NDVI; TMP with MAP and TMP with AET were the main interactive factors in the spatial distribution of the NDVI. (3) The article quantifies the impact of the anthropogenic drivers on land degradation. Human activities positively influenced the NDVI in 91.02% of the area and negatively influenced it in 8.98% of the area. (4) The overall trend of the NDVI was mainly stable, with stronger improvement than degradation, and showed strong persistence. The above findings enrich our understanding of the climatic impacts of land degradation and human impacts in arid or semi-arid regions and provide a scientific basis for ecological engineering to achieve ecological conservation and quality development in Balochistan, Pakistan.

ObjectiveTo explore the clinical efficacy, characteristics and safety of endoscopic-assisted resection of single-segment posterior longitudinal ligament ossification in the treatment of thoracic spinal stenosis (TSS).MethodFifteen TSS patients, including 6 males and 9 females aged 43–70 years treated with endoscopic-assisted resection of single-segment posterior longitudinal ligament ossification through the transfacet joint approach by our team from November 2016 to June 2020 were retrospectively analyzed. The operation time, intraoperative blood loss, and postoperative complications were recorded. The VAS score, ODI and JOA score (full score, 11 points) were recorded before the operation, after the operation and at the last follow-up to evaluate the clinical efficacy and calculate the improvement rate.ResultsThe ventral side of the spinal cord was decompressed in all patients, providing improvements in neurological symptoms and significant pain relief. The mean follow-up time was 20.27 ± 3.87 months. Mean operation time, intraoperative blood loss, and hospitalization time were found to be 84.80 ± 13.23 min, 36.33 ± 7.41 mL, 5.13 ± 1.02 days; respectively.The JOA score at the last follow-up was 8.6 ± 1.25, which was significantly better than the preoperative (5.53 ± 1.20) and postoperative (6.87 ± 1.31) scores (p

Zeolites and zeo-type materials with nanosized dimensions are of great practical interest owing to their favorable transport properties, faster adsorption kinetics, and large external surface area. This mini-review presents recent developments in the organic template-free synthesis of nanosized zeolites and related materials. The advantages and challenges of these methods are addressed with particular attention to the green synthesis of nanozeolites.

Angiokinases, such as vascular endothelial-, fibroblast- and platelet-derived growth factor receptors (VEGFRs, FGFRs and PDGFRs) play crucial roles in tumor angiogenesis. Anti-angiogenesis therapy using multi-angiokinase inhibitor has achieved great success in recent years. In this study, we presented the design, synthesis, target identification, molecular mechanism, pharmacodynamics (PD) and pharmacokinetics (PK) research of a novel triple-angiokinase inhibitor WXFL-152. WXFL-152, identified from a series of 4-oxyquinoline derivatives based on a structure–activity relationship study, inhibited the proliferation of vascular endothelial cells (ECs) and pericytes by blocking the angiokinase signals VEGF/VEGFR2, FGF/FGFRs and PDGF/PDGFRβ simultaneously in vitro. Significant anticancer effects of WXFL-152 were confirmed in multiple preclinical tumor xenograft models, including a patient-derived tumor xenograft (PDX) model. Pharmacokinetic studies of WXFL-152 demonstrated high favourable bioavailability with single-dose and continuous multi-dose by oral administration in rats and beagles. In conclusion, WXFL-152, which is currently in phase Ib clinical trials, is a novel and effective triple-angiokinase inhibitor with clear PD and PK in tumor therapy.

The influenza A virus is highly contagious and often causes global pandemics. The prevalence of strains of the influenza A virus that are resistant to approved drugs is a huge challenge for the current clinical treatment of influenza A. RNA polymerase is a pivotal enzyme in the replication of the influenza A virus, and it is a promising target for anti-influenza A therapies. In this paper, we report a novel and potent anti-influenza-A-virus inhibitor, ZSP1273, targeting the influenza A virus RNA polymerase, especially for multidrug-resistant strains. The inhibitory activity of ZSP1273 on RNA polymerase activity was 0.562 ± 0.116 nM (IC50 value), which was better than that of the clinical candidate compound VX-787 with the same target. In vitro, the EC50 values of ZSP1273 on normal influenza A virus strains (i.e., H1N1 and H3N2) varied from 0.01 nM to 0.063 nM, which were better than those of the licensed drug oseltamivir. Moreover, oseltamivir-resistant strains, baloxavir-resistant strains, and highly pathogenic avian influenza strains were also sensitive to ZSP1273. In vivo, ZSP1273 effectively reduced influenza A virus titers in a dose-dependent manner in a murine model and maintained a high survival rate in mice. In addition, the inhibitory activity of ZSP1273 on influenza A virus infection was also observed in a ferret model. Pharmacokinetic studies showed the favorable pharmacokinetic characteristics of ZSP1273 in mice, rats, and beagle dogs after single-dose and continuous multiple-dose administration. In conclusion, ZSP1273 is a highly effective anti-influenza A virus replication inhibitor, especially against multidrug-resistant strains. ZSP1273 is currently being studied in phase III clinical trials.

It is easy to skid and roll by centrifugal force and wind, which affects the safety of the vehicle. At the same time, the special location of bridge and tunnel connecting segment in the canyon will accelerate the wind, and make the transverse wind play a more important role in driving vehicles. Based on the acceleration effect of canyons on wind, the vehicle model is simulated by Carsim software, and the road and wind models are established. It is studied that the vehicle has different radius of circular curve under different wind levels, corresponding to different superelevation. The lateral acceleration, lateral deflection and transverse force coefficient are selected as the running state of the vehicle at different speeds to research the linear optimization design of bridge and tunnel connecting segment in the canyon. The result shows that when the wind force exceeds the fifth grade, it is possible for the vehicle to overturn under the limit minimum radius required by the standard value. In order to ensure the stability and safety of the vehicle, this paper considers the geographical position of bridge and tunnel connecting segment in the canyon and the relationship between the radius of the superelevation and circular curve. What’s more, it puts forward the optimal limit minimum radius of the circular curve with different wind grades of 5-9 grades. At the same time, when the road alignment cannot be optimized under the condition, this paper puts forward the speed limit that the vehicle safety can be guaranteed under different wind speed conditions. The speed limit can provide reference for traffic management and safety guarantee of mountain expressway.

Sepsis is an exacerbated inflammatory reaction induced by severe infection. As important defensive molecules in innate immunity, several AMPs are reported to prevent septic shock. In this study, we characterized a novel cathelicidin, FM-CATH, from the frog skin of F. multistriata. FM-CATH was found to adopt an amphipathic α-helix structural in membrane-mimetic environments and possess favorable antimicrobial effects against bacteria and fungus. In addition, it triggered the agglutination of bacteria. It could also strongly bind to LPS and LTA. Additionally, FM-CATH affected the enzymatic activities of thrombin, plasmin, β-tryptase, and tPA, leading to coagulation inhibition in vitro and in vivo. Finally, we observed that FM-CATH improved survival rate and inhibited pathological alteration, bacterial count, serum biochemistry, and pro-inflammatory cytokine expression in the cecal ligation and puncture-induced sepsis mice. Taken together, these findings suggest that FM-CATH might be served as a promising agent for the treatment of sepsis.

In this study, systematic separation and subsequent pharmacological activity studies were carried out to identify cytotoxic natural products from the dried stems of Millettia pachyloba Drake. Five previously undescribed isoflavones, pachyvones A–E; one previously undescribed xanthone, pachythone A; and twenty-two known compounds were obtained. The structures of these compounds were assigned on the basis of 1D/2D NMR data and high-resolution electrospray ionization mass spectroscopy analysis. Preliminary activity screening with HeLa and MCF-7 cells showed that ten compounds (3–5, 9, 12, 17–19, 24, and 25) had potential cytotoxicity. Further in-depth activity studies with five cancer cell lines (HeLa, HepG2, MCF-7, Hct116, and MDA-MB-231) and one normal cell line (HUVEC) revealed that these ten compounds showed specific cytotoxicity in cancer cells, with IC50 values ranging from 5 to 40 μM, while they had no effect on normal cell lines. To investigate whether the cytotoxicity of these ten compounds was associated with autophagy, their autophagic effects were evaluated in GFP-LC3-HeLa cells. The results demonstrated that compound 9 (durmillone) significantly induced autophagy in a concentration-dependent manner and had the best activity as an autophagy inducer among all of the compounds. Therefore, compound 9 was selected for further study. The PI/Annexin V double staining assay and Western blotting results revealed that compound 9 also induced obvious apoptosis in HeLa and MCF-7 cells, which suggests that it mediates cytotoxic activity through activation of both apoptosis and autophagy. Taken together, this study identified ten natural cytotoxic products from the dried stems of Millettia pachyloba Drake, of which compound 9 induced apoptosis and autophagy and could be an anticancer drug candidate. Keywords: Millettia pachyloba, Leguminosae, Isoflavones, Cytotoxicity, Autophagy, Apoptosis

Periplaneta americana is a common traditional Chinese medicinal material which has been used to treat arthritis, fever, aches, pains, and inflammation of the extremities for several hundred years. However, little scientific data exists in literature to support its use. The purpose of this study was to evaluate the antipyretic, anti-inflammatory and analgesic activities of Periplaneta americana extract (PAE) and explore its underlying mechanism. The antipyretic, anti-inflammatory and analgesic activities were evaluated by LPS-induced fever, carrageenan-induced paw edema, abdominal writhing, hot plate and formalin tests, respectively. The mechanism of action was explored by antioxidant activity analysis, inflammatory cytokines expression and febrile mediator measurement, and pathway activation analysis. The results from UHPLC-HRMS indicated that the extract was found to contain dopamine, coumarin, dipeptide, vitamin, organic acid, amino acid and its metabolites, and other organic compounds. PAE showed in a dose-dependent manner antioxidant activity and reduced the protein production and mRNA expression of NO, IL-1β, IL-6, and TNF-α in RAW 264.7 cells in vitro. Moreover, PAE significantly and dose-dependently inhibited the writhing responses and licking time in formalin tests, increased response latency in the hot plate test, reduced carrageenan-induced paw edema and inflammation in mice, decreased LPS-induced rT increase in rats. Furthermore, PAE treatment markedly inhibited the increase in the levels of NO, IL-6, IL-1β, TNF-α, PGE2 and cAMP in plasma of fevered rat, greatly suppressed the activation of inflammatory response pathway and the change of MDA and GSH concentration, MPO and SOD activity as well as FRAP capacity in paw induced by carrageenan injection. In conclusion, the findings suggested that PAE produced potential antinociceptive, anti-inflammatory and antipyretic effects by reducing production of endogenous inflammatory mediators and blocking the MAPK/NF-κB signaling pathway which support the claim for its traditional use in the treatment of various diseases.

The impact of fermented cow milk on gut microbiota has been extensively studied, however, the role of fermented goat milk (FGM) has not been well evaluated. The present study using 16S rRNA gene sequencing technology investigated the effect of FGM intake on small intestinal microbiota in mice. It was found that FGM intake decreased microbial diversity and significantly altered microbiota composition, particularly the genera Lactobacillus and Streptococcus. Moreover, metabolism and disease profiles were shifted by the FGM based on the function prediction. The expression of immune factors including tumor necrosis factor alpha (Tnfα), granzyme B (Gzmb), perforin (Prf) and aryl hydrocarbon receptor (Ahr) was also regulated by FGM treatment. Collectively, our findings reveal that FGM intake can exert beneficial effects on small intestinal microbiota and further modulate the host metabolism and immune responses. Therefore, the FGM can serve as an alternative functional food to improve health through gut bacterial management.

Peroxisome proliferator-activated receptor α/δ (PPAR α/δ), regulating glucolipid metabolism and immune inflammation, has been identified as an effective therapeutic target in non-alcoholic steatohepatitis (NASH). Dual PPAR α/δ agonist, such as GFT505 (also known as elafibranor), demonstrated potential therapeutic effect for NASH in clinical trials. To profile the regulatory network of PPAR α/δ agonist in NASH, the choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) induced NASH model was used to test the pharmacodynamics and transcriptome regulation of GFT505 in this study. The results showed that GFT505 ameliorated hepatic steatosis, inflammation and fibrosis in CDAHFD mice model. RNA-sequencing yielded 3995 up-regulated and 3576 down-regulated genes with GFT505 treatment. And the most significant differentialy expressed genes involved in glucolipid metabolism (Pparα, Acox1, Cpt1b, Fabp4, Ehhadh, Fabp3), inflammation (Ccl6, Ccl9, Cxcl14) and fibrosis (Timp1, Lamc3, Timp2, Col3a1, Col1a2, Col1a1, Hapln4, Timp3, Pik3r5, Pdgfα, Pdgfβ, Tgfβ1, Tgfβ2) were confirmed by RT-qPCR. The down-regulated genes were enriched in cytokine-cytokine receptor interaction pathway and ECM-receptor interaction pathway, while the up-regulated genes were enriched in PPAR signaling pathway and fatty acid degradation pathway. This study provides clues and basis for further understanding on the mechanism of PPAR α/δ agonist on NASH.

When a car is running at high speed, the canyon wind at the bridge-tunnel junction in the mountainous area brings along the acceleration effect. The aerodynamic lateral force will cause the vehicle sideslip and unsteady steering, which is extremely harmful to driving safety. In this paper, Xi-Han Expressway is taken as the research object to analyze the influencing factors of vehicle’s lateral stability by combining the theoretical research of the finite element method, automobile aerodynamics, and speed limit with field investigation and simulation test. CarSim software is used for simulation to explore the influence of different positions of the circular curve on vehicle lateral stability. The results show that the wind level affects the tunnel exit’s unfavorable section on the circular curve. The larger the wind level, the larger the proportion of the tunnel exit’s unfavorable section on the circular curve. The proportions of tunnel exit’s unfavorable section on the circular curve under 6–9 wind levels are 33.33%, 38.89%, 55.56%, and 66.67%, respectively. In addition, the lateral stability of vehicles under level 6–8 wind scale is the worst when the tunnel exit is located at 5° position on the circular curve. The results indicate the influence of strong wind on the lateral stability of vehicles in mountainous expressway. The research can optimize the design of the highway tunnel group and provide the basic theory and method basis for the quantitative management and scientific management of the road traffic management department.

Background Non-alcoholic steatohepatitis (NASH) is a progressive liver disease characterized by hepatic steatosis, lobular inflammation and fibrosis. Acetyl-CoA carboxylase (ACC) isoform 1 and 2 involved in de novo lipogenesis (DNL) and fatty acid oxidation have been identified as a therapeutic target in NASH. GS-0976, the inhibitor of ACC1 and ACC2, has achieved favorable therapeutic effects in clinical trials with NASH. The purpose of this study was to explore the transcriptional alterations regulated by GS-0976 in NASH. Methods C57BL/6 mice were fed on a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) or normal diet for 12 weeks. Mice were treated with or without GS-0976 (3 mg/kg per day) in the last 8 weeks. Oil Red O, Haematoxylin-eosin (H & E), and Sirius Red were used to evaluate hepatic steatosis, inflammation and fibrosis. The comparative RNA-sequencing was conducted to analyse the hepatic gene expression profiles in mice. Reverse transcription–polymerase chain reaction analysis was performed to validate the differential expression of representative genes. Results GS-0976 attenuated the steatosis, inflammation, and fibrosis of NASH in CDAHFD mouse model. High-throughput sequencing and differential gene expression analysis showed that there were 516 up-regulated genes and 525 down-regulated genes after GS-0976 treatment. Genes involved in the metabolic process, extracellular matrix formation, immune response, and angiogenesis were significantly enriched. The “Metabolic pathways” and “ECM-receptor interaction” pathways were the most significantly enriched KEGG pathways in the up-regulated and down-regulated differentially expressed genes (DEGs), respectively. Conclusions Transcriptome analysis showed that GS-0976 could regulate the expression of genes related to metabolism, inflammation and fibrosis in NASH. The global transcriptomic changes in gene expression promote the further understanding for the inhibition mechanisms of GS-0976 in NASH.

Colorectal cancer (CRC) is still a big health burden worldwide. Nutrition and dietary factors are known to affect colorectal cancer development and prognosis. The protective roles of diets rich in fruits and vegetables have been previously reported to contain high levels of cancer-fighting phytochemicals. Anthocyanins are the most abundant flavonoid compounds that are responsible for the bright colors of most blue, purple, and red fruits and vegetables, and have been shown to contribute to the protective effects of fruits and vegetables against cancer and other chronic diseases. Berries and grapes are the most common anthocyanin-rich fruits with antitumor effects. The antitumor effects of anthocyanins are determined by their structures and bioavailability as well as how they are metabolized. In this review, we aimed to discuss the preventive as well as therapeutic potentials of anthocyanins in CRC. We summarized the antitumor effects of anthocyanins and the mechanisms of action. We also discussed the potential pharmaceutical application of anthocyanins in practice.

Interocular suppression was quantified by the interocular luminance difference that was needed when the two eyes were balanced in discriminating a black–white stripe formed butterfly stimulus, which was dichoptically presented through polarized glasses. Stronger interocular suppression was found in amblyopes than that in controls at both the near (33 cm, 0.95 ± 1.00 vs. 0.14 ± 0.18, p

How to precisely quantify the binocular eye balance (i.e., the contribution that each eye makes to the binocular percept) across a range of spatial frequencies using a binocular combination task, is an important issue in both clinical and basic research. In this study, we aimed to compare the precision of a binocular orientation combination paradigm with that of the standard binocular phase combination paradigm in measuring the binocular eye balance at low to high spatial frequencies. Nine normal adults (average age: 24.6 ± 2.0 years old) participated. Subjects viewed an LED screen dichoptically with polarized glasses in a dark room. The method of constant stimuli was used to quantitatively assess the point of subjective equality (PSE), i.e., the interocular contrast ratio when two eyes are balanced in binocular combination, for stimulus spatial frequencies from 0.5 to 8 cycles/degree. Precision was quantified by the variance [i.e., standard error (SE), obtained from 100 bootstrap estimates] associated to the PSE. Using stimuli whose interocular phase difference at the edge of the gratings was matched at 45°, we found that the orientation paradigm provides more precision than the standard binocular phase combination paradigm, especially at high frequencies (Experiment 1). Such differences remained when using stimuli that had three times larger interocular phase difference (Experiment 2) or displayed at four times higher stimuli resolution (Experiment 3). Our results indicate that a binocular combination tasked based on orientation rather than phase, provides a more precise estimate of binocular eye balance in human adults at high spatial frequencies, thus allowing a binocular balance to be assessed within the spatial region where amblyopes are most defective (i.e., high spatial frequencies).

Recent laboratory findings suggest that short-term patching of the amblyopic eye (i.e., inverse occlusion) results in a larger and more sustained improvement in the binocular balance compared with normal controls. In this study, we investigate the cumulative effects of the short-term inverse occlusion in adults and old children with amblyopia. This is a prospective cohort study of 18 amblyopes (10-35 years old; 2 with strabismus) who have been subjected to 2 hours/day of inverse occlusion for 2 months. Patients who required refractive correction or whose refractive correction needed updating were given a 2-month period of refractive adaptation. The primary outcome measure was the binocular balance which was measured using a phase combination task; the secondary outcome measures were the best-corrected visual acuity which was measured with a Tumbling E acuity chart and converted to logMAR units and the stereoacuity which was measured with the Random-dot preschool stereogram test. The average binocular gain was 0.11 in terms of the effective contrast ratio (z=−2.344, p=0.019, 2-tailed related samples Wilcoxon Signed Rank Test). The average acuity gain was 0.13 logMAR equivalent (t17=4.76, p

Partial discharge (PD) detection has been proved as an effective tool for insulation condition monitoring of power equipment. The energy of PD pulses is valuable for studying the characteristics of PD activities. This paper proposes a method for estimating the energy of PD pulses in the presence of white noise and narrowband noise. First, a maximum likelihood (ML) estimator of the pulse energy is derived from the probability distribution of the energy spectral coefficients. To implement the ML method, the sampled data are divided into signal frames and noise frames. The noise frames are then utilized for extracting noise parameters using the 3F-C method. Eventually, these noise parameters are applied to the signal frames to find the ML estimate of the pulse energy. To verify the effectiveness of the proposed method, both simulated data and measured data have been processed using the proposed method and the conventional wavelet packet (WP) denoising method. Compared with the WP denoising method, the proposed method has a higher accuracy and is less susceptible to the lengths of the sampling time windows. The advantage of the method is more significant in unfavorable conditions where the signal-to-noise ratio is low and the accurate lengths of the PD pulses are difficult to determine.

Partial discharge (PD) leads to the generation of electrical, acoustic, optical, and thermal signals. The propagation characteristics of optical signals in gas insulated metal-enclosed transmission lines (GIL) are the basis of optical detection research. This paper simulates the propagation of PD optical signals in GIL through modeling GIL with different structures and specification parameters. By analyzing the optical parameters on the probe surface and the detection points when the PD source position is different, the influence of the difference in specifications caused by the voltage level on the propagation of the GIL PD optical signal is studied. The results show that the GIL cavity structure will affect the faculae distribution and the relative irradiance (RI) of the detection surface; the PD source position has a huge impact on the faculae distribution on the detection surface, but has little influence on the RI; as the voltage rises, the faculae distribution on the detection surface becomes more obvious, and the mean of RI decreases. The above results have the reference value for the manufacture of GIL equipment and the research of PD optical detection. When the specular reflection coefficient of surface material is smaller and the diffuse reflection coefficient is larger, the outline of the light spot is clearer, the proportion of brighter parts is larger, and the maximum value of the RI is larger.

Abstract Gene therapy provides a novel method for cancer therapy. This study shows a DNA nanocomplex that is inspired from vesicular stomatitis virus (VSV) for ovarian cancer therapy. This DNA nanocomplex consists of a cationized monomethoxy poly (ethylene glycol)‐poly (d,l‐lactide) (MPEG‐PLA) nanoparticle and a plasmid encoding the matrix protein of vesicular stomatitis virus (VSVMP) that plays a critical role in the VSV‐induced apoptosis of cancer cells. The cationized MPEG‐PLA nanoparticle that is self‐assembled by MPEG‐PLA copolymer and N‐[1‐(2,3‐dioleoyloxy) propyl]‐N,N,N‐trimethylammonium chloride (DOTAP) has low cytotoxicity and high transfection efficiency (>80%). Intraperitoneal administration of the pVSVMP nanocomplex remarkably inhibits the intraperitoneal metastasis of ovarian cancer and does not cause significant systemic toxicity. The apoptosis induction and anti‐angiogenesis are involved in the anticancer mechanism. This work demonstrates a VSV‐inspired DNA nanocomplex that has potential application for the treatment of intraperitoneal metastasis of ovarian cancer.

Mai Tong Fang (MTF), a Chinese herbal combination, has been used for the treatment of diabetic nephropathy in traditional medical clinics in China. However, the anti-adipogenic and anti-hyperglycemic effects of MTF have not been fully elucidated, so this study explored these pharmacological activities in 3T3-L1 adipocytes and ob/ob mice, respectively, of the water fraction of milkvetch root, salviae miltiorrhizae and mulberry as key components of MTF. MTF was found to inhibit adipogenesis and triglyceride accumulation in 3T3-L1 adipocytes. Oral administration of MTF in ob/ob mice for 8 weeks, exhibited positive controls on blood glucose and body weight, and further improved glucose tolerance according to an oral glucose tolerance test. Importantly, MTF extract alleviated fat deposition and ballooning degeneration in liver tissue and blocked the increase of adipocyte size in adipose tissue from treated ob/ob mice. These results indicated that the extract of key components in the traditional Chinese prescription MTF continue a potent anti-adipogenic and glucose-lowering agent.

For 110 kV and above tunnel-installed high-voltage (HV) cross-linked poly-ethylene (XLPE) cable systems, it is a normal procedure to adopt a cross-bonding scheme. The high-frequency current method is frequently used in the cross-bonded cable systems for on-site or online partial discharge (PD) detection by monitoring the signals on the cross-bonding wires. To further study the amplitude distribution characteristics of the PD signals, a parametric characteristic admittance model of a three-phase cable system in a tunnel is established based on Tylavsky’s formulas. The model is used to calculate the amplitude distribution formula of the PD pulse current on the cross-bonding wires. In addition, the influence of cable laying and tunnel environment on the amplitude distribution is also studied. Finally, the correctness of the model and the conclusion are verified by simulation experiments and on-site tests. The results show that the signal amplitude distribution is determined by the ratio of the characteristic admittances. As the distance between the cables and the distance from the inner wall of the tunnel increase, the amplitude difference gradually decreases.

Exfoliative cytology has been widely used for early diagnosis of oral squamous cell carcinoma (OSCC). Test outcome is reported as "negative", "atypical" (defined as abnormal epithelial changes of uncertain diagnostic significance), and "positive" (defined as definitive cellular evidence of epithelial dysplasia or carcinoma). The major challenge is how to properly manage the "atypical" patients in order to diagnose OSCC early and prevent OSCC. In this study, we collected exfoliative cytology data, histopathology data, and clinical data of normal subjects (n=102), oral leukoplakia (OLK) patients (n=82), and OSCC patients (n=93), and developed a data analysis procedure for quantitative risk stratification of OLK patients. This procedure involving a step called expert-guided data transformation and reconstruction (EdTAR) which allows automatic data processing and reconstruction and reveals informative signals for subsequent risk stratification. Modern machine learning techniques were utilized to build statistical prediction models on the reconstructed data. Among the several models tested using resampling methods for parameter pruning and performance evaluation, Support Vector Machine (SVM) was found to be optimal with a high sensitivity (median>0.98) and specificity (median>0.99). With the SVM model, we constructed an oral cancer risk index (OCRI) which may potentially guide clinical follow-up of OLK patients. One OLK patient with an initial OCRI of 0.88 developed OSCC after 40 months of follow-up. In conclusion, we have developed a statistical method for qualitative risk stratification of OLK patients. This method may potentially improve cost-effectiveness of clinical follow-up of OLK patients, and help design clinical chemoprevention trial for high-risk populations.

Mammalian esophagus exhibits a remarkable change in epithelial structure during the transition from embryo to adult. However, the molecular mechanisms of esophageal epithelial development are not well understood. Zebrafish (Danio rerio), a common model organism for vertebrate development and gene function, has not previously been characterized as a model system for esophageal epithelial development. In this study, we characterized a piece of non-keratinized stratified squamous epithelium similar to human esophageal epithelium in the upper digestive tract of developing zebrafish. Under the microscope, this piece was detectable at 5dpf and became stratified at 7dpf. Expression of esophageal epithelial marker genes (Krt5, P63, Sox2 and Pax9) was detected by immunohistochemistry and in situ hybridization. Knockdown of P63, a gene known to be critical for esophageal epithelium, disrupted the development of this epithelium. With this model system, we found that Pax9 knockdown resulted in loss or disorganization of the squamous epithelium, as well as down-regulation of the differentiation markers Krt4 and Krt5. In summary, we characterized a region of stratified squamous epithelium in the zebrafish upper digestive tract which can be used for functional studies of candidate genes involved in esophageal epithelial biology.

Morphological changes during human and mouse esophageal development have been well characterized. However, changes at the molecular level in the course of esophageal morphogenesis remain unclear. This study aims to globally profile critical genes and signaling pathways during the development of mouse esophagus. By using microarray analysis this study also aims to determine how the Nrf2/Keap1 pathway regulates the morphogenesis of the esophageal epithelium.Gene expression microarrays were used to survey gene expression in the esophagus at three critical phases: specification, metaplasia and maturation. The esophagi were isolated from wild-type, Nrf2(-/-), Keap1(-/-), or Nrf2(-/-)Keap1(-/-) embryos or young adult mice. Array data were statistically analyzed for differentially expressed genes and pathways. Histochemical and immunohistochemical staining were used to verify potential involvement of the Wnt pathway, Pparβ/δ and the PI3K/Akt pathway in the development of esophageal epithelium.Dynamic gene expression patterns accompanied the morphological changes of the developing esophagus at critical phases. Particularly, the Nrf2/Keap1 pathway had a baseline activity in the metaplasia phase and was further activated in the maturation phase. The Wnt pathway was active early and became inactive later in the metaplasia phase. In addition, Keap1(-/-) mice showed increased expression of Nrf2 downstream targets and genes involved in keratinization. Microarray and immunostaining data also suggested that esophageal hyperkeratosis in the Keap1(-/-) mice was due to activation of Pparβ/δ and the PI3K/Akt pathway.Morphological changes of the esophageal epithelium are associated with dynamic changes in gene expression. Nrf2/Keap1 pathway activity is required for maturation of mouse esophageal epithelium.