Reproductive Ability Disparity in the Pacific Whiteleg Shrimp (Penaeus vannamei): Insights from Ovarian Cellular and Molecular Levels.

The Pacific whiteleg shrimp (Penaeus vannamei) is a highly significant species in shrimp aquaculture. In the production of shrimp larvae, noticeable variations in the reproductive capacity among female individuals have been observed. Some females experience slow gonadal development, resulting in the inability to spawn, while others undergo multiple maturations and contribute to the majority of larval supply. Despite numerous studies that have been conducted on the regulatory mechanisms of ovarian development in shrimp, the factors contributing to the differences in reproductive capacity among females remain unclear. To elucidate the underlying mechanisms, this study examined the differences in the ovarian characteristics between high and low reproductive bulks at different maturity stages, focusing on the cellular and molecular levels. Transmission electron microscopy analysis revealed that the abundance of the endoplasmic reticulum, ribosomes, mitochondria, and mitochondrial cristae in oocytes of high reproductive bulk was significantly higher than that of the low reproductive bulk in the early stages of ovarian maturation (stages I and II). As the ovaries progressed to late-stage maturation (stages III and IV), differences in the internal structures of oocytes between females with different reproductive capacities gradually diminished. Transcriptome analysis identified differentially expressed genes (DEGs) related to the mitochondria between two groups, suggesting that energy production processes might play a crucial role in the observed variations in ovary development. The expression levels of the ETS homology factor (EHF) and PRDI-BF1 and RIZ homology domain containing 9 (PRDM9), which were significantly different between the two groups, were compared using qRT-PCR in individuals at different stages of ovarian maturation. The results showed a significantly higher expression of the EHF gene in the ovaries of high reproductive bulk at the II and IV maturity stages compared to the low reproductive bulk, while almost no expression was detected in the eyestalk tissue of the high reproductive bulk. The PRDM9 gene was exclusively expressed in ovarian tissue, with significantly higher expression in the ovaries of the high reproductive bulk at the four maturity stages compared to the low reproductive bulk. Fluorescence in situ hybridization further compared the expression patterns of EHF and PRDM9 in the ovaries of individuals with different fertility levels, with both genes showing stronger positive signals in the high reproductive bulk at the four ovarian stages. These findings not only contribute to our understanding of the regulatory mechanisms involved in shrimp ovarian development, but also provide valuable insights for the cultivation of new varieties aimed at improving shrimp fecundity.

Reliability-based anti-disturbance control for systems with parametric stochastic uncertainty: A probabilistic LMI approach.

We propose a reliability-based anti-disturbance control (RADC) method for systems with parametric stochastic uncertainty based on the linear matrix inequality (LMI) and the limit state function. Differing from the existing anti-disturbance control, the parametric stochastic uncertainty is considered in both the concerned system and the exogenous disturbance system. With this consideration, the condition for system stability and performance robustness is described by a stochastic LMI which holds with a certain probability (reliability). Through the limit state function method, the stochastic LMI is subtly transformed into two probabilistic LMIs for two different cases. The proposed probabilistic LMIs contain two probabilistic parameters of reliability indexes that quantify the effect of parametric stochastic uncertainty. At different prescribed reliability indexes, controllers with different reliability can be flexibly and reliably designed. Two illustrative examples with Monte-Carlo verification are presented to demonstrate the feasibility and effectiveness of the proposed RADC method.

Survival benefit of local consolidative therapy for patients with single-organ metastatic pancreatic cancer: a propensity score-matched cross-sectional study based on 17 registries.

Background: The continuous exploration of oligometastatic disease has led to the remarkable achievements of local consolidative therapy (LCT) and favorable outcomes for this disease. Thus, this study investigated the potential benefits of LCT in patients with single-organ metastatic pancreatic ductal adenocarcinoma (PDAC). Methods: Patients with single-organ metastatic PDAC diagnosed between 2010 - 2019 were identified from the Surveillance, Epidemiology, and End Results (SEER) database. Propensity score matching (PSM) was performed to minimize selection bias. Factors affecting survival were assessed by Cox regression analysis and Kaplan-Meier estimates. Results: A total of 12900 patients were identified from the database, including 635 patients who received chemotherapy combined with LCT with a 1:1 PSM with patients who received only chemotherapy. Patients with single-organ metastatic PDAC who received chemotherapy in combination with LCT demonstrated extended median overall survival (OS) by approximately 57%, more than those who underwent chemotherapy alone (11 vs. 7 months, p < 0.001). Furthermore, the multivariate Cox regression analysis revealed that patients that received LCT, younger age (< 65 years), smaller tumor size (< 50 mm), and lung metastasis (reference: liver) were favorable prognostic factors for patients with single-organ metastatic PDAC. Conclusion: The OS of patients with single-organ metastatic pancreatic cancer who received LCT may be prolonged compared to those who received only chemotherapy. Nevertheless, additional prospective randomized clinical trials are required to support these findings.

Combining a protein-targeting small molecule and a thiol-targeting small molecule for detecting a serum risk marker of liver tumor recurrence.

This work proposes a novel bioassay designed to detect the 2B receptor of serotonin in serum samples, which can serve as a risk marker for cancer recurrence after surgical resection. Traditional methods for detecting this marker are often costly and time-consuming, requiring specialized reagents and equipment. The new bioassay is designed to enable direct and reagent-less detection of the 2B receptor in serum samples, without the need of antibodies or enzymes. The assay uses a small molecule ligand for the 2B receptor combined with a thiol-targeting fluorescent dye on a compact peptide-based molecular frame. This design allows for a rapid and specific readout of the fluorescent signal upon probe-protein interaction. In addition, the covalent biosensing process used in the assay allows for signal enhancement by electrochemical cross-linking of serum proteins. The bioassay was successfully used to detect the 2B receptor in serum samples from hepatocarcinoma patients, indicating its potential as a powerful tool for early cancer detection and monitoring.

Multiplicative fault concept and fault observer design for reliability study of degrading reaction wheel.

The reaction wheel is a typical actuator and a crucial weak link in the spacecraft attitude control system, and much attention has been paid to its reliability problem. Due to limited samples and high cost for reaction wheel life tests, a simulation method by introducing attitude coupling dynamics and multiplicative fault concept is developed to analyze the logic of electric current as a performance indicator and verify its accuracy for reliability modeling. Furthermore, a new and intrinsic performance indicator of multiplicative fault is proposed for more application scenarios of reliability modeling and an adaptive sliding mode observer is designed for fault estimation. An illustrative example shows that the performance indicator of multiplicative fault can be used for various mission scenarios but requires certain persistent excitation, while electric current is the opposite.

A novel peptide-templated AgNPs nanoprobe for theranostics of prostate cancer.

Prostate specific membrane antigen (PSMA)-positive exosomes have the potential to serve as highly sensitive biomarkers for prostate cancer detection. Herein, a sensitive electrochemical biosensor for the ultrasensitive detection of PSMA-positive exosomes has been constructed based on a peptide-templated AgNPs nanoprobe. In this work, PSMA-specific binding peptides immobilized on a gold electrode were responsible for prostate cancer-derived exosomes capturing. Well-designed peptide (CCY- LWYIKC) serves a dual role: as a signal probe and as a recognizer in the exosomes-identification process. Specifically, LWYIKC bind to cholesterol at the exosome membranes, and CCY function as peptide templates to host a large number of silver nanoparticles, leading to a strong electrochemical signal. Thus, the concentration of exosomes can be quantified via electrochemical signal. This innovative method displayed a wide detection range of 102 to 108 particles/µL and a detection limit as low as 37 particles/µL. Notably, the method has shown outstanding performance when validated using clinical samples, suggesting its potential for clinical applications.

Regeneration mechanism of a novel high-performance biochar mercury adsorbent directionally modified by multimetal multilayer loading.

Biochar that is directly obtained by pyrolysis exhibits a low adsorption efficiency; furthermore, the process of recycling adsorbents is ineffective. To solve these problems, conventional chemical coprecipitation, sol-gel, multimetal multilayer loading and biomass pyrolysis coking processes have been integrated. After selecting specific components for structural design, a novel high-performance biochar adsorbent was obtained. The effects of the O2 concentration and temperature on the regeneration characteristics were explored. An isothermal regeneration method to repair the deactivated adsorbent in a specific atmosphere was proposed, and the optimal regeneration mode and conditions were determined. The microscopic characteristics of the regenerated samples were revealed along with the mechanism of Hg0 removal and regeneration by using temperature-programmed desorption technology and adsorption kinetics. The results show that doping multiple metals can reduce the pyrolysis reaction barrier of the modified biomass. On the modified surface of the sample, the doped metals formed aggregated oxides, and the resulting synergistic effect enhanced the oxidative activity of the biochar carriers and the threshold effect of Ce oxide. The optimal regeneration conditions (5% O2 and 600 °C) effectively coordinated the competitive relationship between the deep carbonization process and the adsorption/oxidation site repair process; in addition, these conditions provided outstanding structure-effect connections between the physico-chemical properties and Hg0 removal efficiency of the regenerated samples. Hg0 adsorption by the regenerated samples is a multilayer mass transfer process that involves the coupling of physical and chemical effects, and the surface adsorption sites play a leading role.

Factors associated with psychological resilience in patients with chronic heart failure and efficacy of psycho-cardiology intervention.

OBJECTIVE: To explore the value of psycho-cardiology intervention on psychological resilience in patients with chronic heart failure (CHF) and investigate the associated factors. METHODS: A retrospective study of 142 patients with CHF was carried out. These patients were admitted to the Department of Cardiology, Provincial Clinical Medical College of Fujian Medical University from January 2017 to January 2021. They were grouped according to intervention method, including 74 patients with psycho-cardiology intervention and 68 with conventional intervention. The psychological resilience and the levels of anxiety and depression before and after intervention were assessed with the Connor-Davidson resilience scale (CD-RISC), self-rating anxiety scale (SAS), and self-rating depression scale (SDS), respectively. The factors associated with psychological resilience in patients with CHF were observed. The relationship between psychological resilience and SAS scores before intervention was studied. RESULTS: Using multivariate logistic regression analysis, we found that age (OR (95% CI): 3.452 (0.862-4.872), P=0.015), gender (OR, (95% CI): 3.389 (0.872-5.023), P=0.035), SAS score (OR (95% CI) 5.433 (1.543-14.333), P=0.027) and SDS score (OR (95% CI): 5.654 (1.572-15.823), P=0.021) were factors associated with psychological resilience in patients with CHF (all P<0.05). The average CD-RISC scores were 56.55±8.89 points in patients with CHF. The psychological resilience was inversely correlated with SAS score (r=-0.450, P<0.001) and SDS scores (r=-0.401, P<0.001). The CD-RISC scores of the observation group after intervention were higher than before intervention and higher than the control group, while SAS and SDS scores were decreased (all P<0.05). CONCLUSION: Age, gender, SAS, and SDS scores are factors associated with psychological resilience in patients with CHF. Psychological resilience was inversely associated with both anxiety and depression. Psycho-cardiology intervention can improve patients' psychological resilience, and reduce their anxiety and depression.

First-Line Tyrosine Kinase Inhibitors Combined With Local Consolidative Radiation Therapy for Elderly Patients With Oligometastatic Non-Small Cell Lung Cancer Harboring EGFR Activating Mutations.

OBJECTIVE: The aim of this study was to investigate the efficacy and safety of combined applications of local consolidative radiation therapy (LCRT) and first-line tyrosine kinase inhibitors (TKIs) for the treatment of primary tumors and oligometastatic sites in oligometastatic NSCLC harboring Epidermal Growth Factor Receptor (EGFR) activating mutations. PATIENTS AND METHODS: Elderly patients with oligometastatic NSCLC (≤5 metastases) harboring EGFR activating mutations at the time of diagnosis were identified. They were treated with first-line TKIs alone or in combination with LCRT. Progression-free survival (PFS) and overall survival (OS) were estimated through the Kaplan-Meier method. RESULTS: A total of 122 elderly patients were enrolled between February 2010 and January 2018. Among them, 41.0% (n = 50) received TKIs combined with LCRT (TKIs + LCRT group), whereas 59.0% (n = 72) received TKIs monotherapy (TKIs alone group). Patients were followed up for a median length of 34 months (ranging from 7.0 to 64 months). The median PFS in TKIs + LCRT group was 17 months (95%CI: 15.37-18.63), which was significantly longer than that of the TKIs-alone group (12 months; 95%CI: 11.05-12.95) (p <0.001). Median OS in TKIs + LCRT group was 38 months (95%CI: 35.61-40.39), while that of the TKIs-alone group was 29 months (95%CI: 26.86-31.14) (p <0.001). Multivariate analyses revealed that LCRT, one to two metastases, and good ECOG PS were independent predictors for better PFS (p <0.001, p = 0.004, and p = 0.027). Moreover, LCRT, good ECOG PS, and T1-2 stage were independent predictors for better OS (p <0.001, p = 0.007 and p = 0.007). Most of the patients suffered from grade 1 to 2 toxicities, and treatment-related deaths were not recorded. CONCLUSION: First-line TKIs combined with LCRT may improve survival outcomes for elderly patients with oligometastatic NSCLC harboring EGFR activating mutations. This approach was not associated with much toxicity, therefore, it can be used for the treatment of elderly patients with oligometastatic disease.

Ultrasensitive fluorescent detection of telomerase activity based on tetrahedral DNA nanostructures as carriers for DNA-templated silver nanoclusters.

Precise evaluation of telomerase activity is essential for the clinical diagnosis of early tumors. Herein, we have ingeniously designed a tetrahedral DNA nanostructure, with hairpin-shaped DNA probes rich in cytosine bases at four vertices for telomerase detection. The DNA-templated silver nanoclusters can be formed after the addition of Ag. Then the introduction of telomerase adds the single-strand TTAGGG extension, which can "turn on" the fluorescence of silver nanoclusters quickly by the proximity of the resulting guanine-rich sequences to silver nanoclusters and realize accurate detection of telomerase activity. In this study, integration of high stability tetrahedral DNA nanostructure and fluorescence signal amplification of four DNA-templated silver nanoclusters offers the advantage of high sensitivity, with a low detection limit of 1 cell. More than that, this method is low-cost, facile, and feasible for practical clinical applications.

Xanthotoxin induced photoactivated toxicity, oxidative stress and cellular apoptosis in Caenorhabditis elegans under ultraviolet A.

Xanthotoxin (XAT) is widely present in many kinds of plants. Caenorhabditis elegans, a typical model organism, was used to study the effects of XAT on C. elegans developmental toxicity, neurotoxicity, reproductive toxicity induced under ultraviolet A (UVA), oxidative stress and apoptosis in C. elegans. The results showed that after XAT exposure treatment, the hatchability of C. elegans decreased significantly as the concentration increased; the body length and width increased markedly, the external morphology was swollen; the brood sizes had been decreased; and the frequencies of head thrashes and body bend decreased significantly. At 80 and 100 mg/L, XAT reduced the activities of mitochondrial complex enzymes I and III, resulting in the excessive production of ROS, and inhibited SOD and CAT so that the ROS could not be eliminated over time. ROS accumulation in the bodies further caused the contents of MDA, protein carbonyl and lipofuscin to increase significantly, the mitochondrial membrane potential to be severely damaged, apoptosis to occur, and the apoptosis genes ced-3 and ced-4 to be significantly upregulated. Thus, XAT showed photoactivated toxicity to C. elegans under UVA, which will help people to make full and rational use of plants containing XAT.

Relationship of anifrolumab pharmacokinetics with efficacy and safety in patients with systemic lupus erythematosus.

OBJECTIVES: To characterize the relationship of anifrolumab pharmacokinetics with efficacy and safety in patients with moderate to severe SLE despite standard therapy, using pooled data from two phase 3 trials. METHODS: TULIP-1 and TULIP-2 were randomized, placebo-controlled, 52-week trials of intravenous anifrolumab (every 4 weeks for 48 weeks). For the exposure-response analysis, BILAG-based Composite Lupus Assessment (BICLA) or SLE Responder Index [SRI(4)] response rates at week 52 in each quartile/tertile of average anifrolumab serum concentration (Cave) were compared for anifrolumab and placebo in all-comers, patients who completed treatment, and IFN gene signature (IFNGS)-high patients who completed treatment, using average marginal effect logistic regression. Relationships between exposure and key safety events were assessed graphically. RESULTS: Of patients in TULIP-1/TULIP-2 who received anifrolumab (150 mg, n = 91; 300 mg, n = 356) or placebo (n = 366), 574 completed treatment, of whom 470 were IFNGS high. In the exposure-efficacy analyses, BICLA and SRI(4) treatment differences favouring anifrolumab 300 mg vs placebo were observed across Cave subgroups and all analysis populations. Logistic regression identified Cave as a significant covariate for predicted BICLA response, as higher anifrolumab Cave predicted greater efficacy. There was no evidence of exposure-driven incidence of key safety events through week 52 in patients receiving anifrolumab 150 or 300 mg. CONCLUSION: While higher Cave predicted greater efficacy, consistent positive benefit favouring anifrolumab 300 mg vs placebo was observed in BICLA and SRI(4) responses across Cave subgroups in the TULIP trials. There was no evidence of exposure-driven safety events. CLINICALTRIAL.GOV NUMBERS: NCT02446912, NCT02446899.

Efficacy and Safety of Local Radiotherapy to All Oligometastatic Sites in Elderly Patients with Metachronous Oligometastatic Cancers After Initial Treatment for the Primary Tumor.

BACKGROUND AND PURPOSE: This study aimed to investigate the efficacy and safety of maintenance therapy combined with local radiotherapy at all oligometastatic sites (LRTOS) in elderly patients with metachronous oligometastatic cancers (MOC). PATIENTS AND METHODS: A total of 242 elderly patients with MOC (≤5 metastases) and primary tumor well controlled after definitive treatment was retrospectively analyzed between August 2014 and February 2020 at Beijing Geriatric Hospital and Air Force General Hospital. Patients were divided into maintenance therapy group (maintenance therapy alone) and local radiotherapy group (maintenance therapy combined with LRTOS). RESULTS: There were 86 patients in the local radiotherapy group and 156 patients in the maintenance therapy group. The median length of follow-up was 36 months (range, 8.0-62 months). Median overall survival (mOS) was 25 months (95% CI: 21.1-28.9) in the local radiotherapy group and 16 months (95% CI: 14.5-17.6) in the maintenance therapy group (p < 0.001). Multivariate analyses demonstrated that LRTOS (hazard ratio (HR) = 0.49, 95% confidence interval (CI): 0.35-0.67, p < 0.001), good Eastern Cooperative Oncology Group Performance Status (ECOG PS, HR = 0.69, 95% CI: 0.49-0.97, p = 0.032), longer duration between diagnosis of primary tumor and occurrence of progression (HR = 0.87, 95% CI: 0.78-0.97, p = 0.015), and subsequent systemic treatment (HR = 0.52, 95% CI: 0.38-0.72, p < 0.001) were independent predictors of good OS. In patients who did not receive subsequent systemic treatment, their mOS was 21 months (95% CI: 12.8-29.2) for those treated with LRTOS and 14 months (95% CI: 11.4-16.6) for those who did not receive local radiotherapy (p = 0.001). Further multivariate analysis showed that LRTOS was the only independent factor for predicting good OS (HR = 0.47, 95% CI: 0.26-0.83, p = 0.010). Patients with metachronous oligometastatic lung cancer, colorectal cancer, prostate cancer, and breast cancer had higher survival benefits following LRTOS. Most patients suffered from grade 1-2 toxicities, but no treatment-related death was recorded. CONCLUSION: This retrospective study shows that elderly patients with MOC treated with LRTOS may have better survival outcomes.

Study on the Hg0 removal characteristics and synergistic mechanism of iron-based modified biochar doped with multiple metals.

An iron-based composite adsorbent with biochar as the support was prepared by coprecipitation and the sol-gel method. Both single-iron-based modified biochar without doping with other metals and iron-based modified biochar doped with multiple metals (Ce, Cu, Co, Mn) were synthesised. The adsorption kinetics were analysed, and temperature-programmed desorption measurements were performed to reveal the inherent difference in mechanism between the oxidation and adsorption of Hg0 by the modified biochar and to elucidate the key mechanism of Hg0 removal. The results show that the removal of Hg0 by the modified biochar mainly includes adsorption and oxidation processes. The adsorption process is divided into two stages, external and internal mass transfer, both of which occur via multilayer adsorption. HgO and Hg-OM are the main forms of Hg0 present on the modified biochar surface. Doped metal oxides can play a synergistic role in enhancing the mercury removal performance of the modified biochar.

Assessing efficacy and safety of stereotactic body radiation therapy for oligometastatic non-small cell lung cancer with epidermal growth factor receptor (EGFR) wild type.

BACKGROUND: Stereotactic body radiation therapy (SBRT) is an emerging therapy for oligometastatic cancer. The aim of this study was to investigate the efficacy and safety of high-dose radiotherapy for primary and oligometastatic lesions in epidermal growth factor receptor (EGFR) wild-type non-small cell lung cancer (NSCLC). METHODS: A total of 40 EGFR wild-type oligometastatic NSCLC patients (defined as ≤5 metastases) treated with SBRT in our department between 2009 and 2016 were analyzed retrospectively. SBRT was delivered to the lesions with a median biologically effective dose at alpha/beta 10 (BED10) value of 102.7 Gy (range, 94.5-113.5 Gy). Primary endpoints including progression-free survival (PFS) and overall survival (OS) were estimated with the Kaplan-Meier method. Factors potentially affecting OS and PFS were evaluated by univariate and multivariate Cox-regression analyses. RESULTS: After a median follow-up of 39 months, the median OS observed in this study was 40 months (95% CI: 32.562-47.438 months). One-, 2-, and 3-year OS rates were 100.0%, 72.5%, and 62.5% respectively. Twenty-nine patients (72.5%) succumbed to tumor burden and median PFS was 13 months (range, 10.687-15.313 months). One-, 2-, and 3-year PFS rates were 65.0%, 10.0%, and 0% respectively. Multivariate analysis suggested Eastern Cooperative Oncology Group performance status (ECOG PS) <2 and high-dose radiation regimens were independent prognostic factors of longer OS (P<0.001 and 0.049, respectively), and patients receiving radiotherapy with BED10 ≥100 Gy showed a better PFS than those undergoing low dose (P=0.047). There were no patients of CTCAE v 5.0 grade 4-5 toxicity or treatment-related deaths. Grade 3 toxicity occurred in 2 (5.0%) patients and 36 (90.0%) patients experienced grade 1-2 adverse reactions. CONCLUSIONS: The current study suggested systemic chemotherapy combined with SBRT for pulmonary and metastatic lesions was feasible and tolerable to improve outcomes of EGFR wild-type oligometastatic NSCLC patients.

A new method for identification of outliers in immunogenicity assay cut point data.

The cut point is an important parameter for immunogenicity assay validation and critical to immunogenicity assessment in clinical trials. FDA (2019) recommends using a statistical approach to derive cut point, with an appropriate outlier removal procedure. In general, the industry follows the methods described in Shankar et al. (2008) and Zhang et al. (2013) among others to determine cut point. Outlier removal is a necessary step during the cut point determination exercise to reduce potential false negative classifications. However, the widely used statistical outlier removal method, namely, Tukey's box-plot method (1.5 times inter-quartile range, IQR), is often found to be overly conservative in the sense that it removes too many "outliers". Tukey's box-plot method can be used to flag potential outliers for further investigation, however, it is not a hypothesis testing based statistical method. Removing these suspected "outliers" will lead to lower cut point which might confound immunogenicity assessment due to the presence of many low false positives. Besides, the very nature of assay analytical variability has a non-negligible adverse impact on the reliability of ADA classification in terms of false positive and false negative, demanding as large as possible contribution from biological variability relative to analytical variability. A new outlier removal procedure, which takes into account the relative magnitude between biological variability and analytical variability within the sample population, is proposed and statistically justified. After sequential removal of analytical and biological outliers, a 5% false positive rate and 1% false positive rate in screening and confirmatory assays, respectively, are still targeted without increasing potential false negatives. Internal data shows that this practice has minimal impact on assay sensitivity and has the advantage of selecting true positive samples. It is shown that the new procedure is more appropriate for cut point determination.

Confirmatory cut point has limited ability to make accurate classifications in immunogenicity assays.

Aim: Competitive inhibition with excess unlabeled drug is used to confirm the presence of antidrug antibodies (ADA) in study samples. We evaluated specific and nonspecific responses from both drug-naive and drug-treated subjects to identify conditions required by the confirmatory assay to make accurate ADA classifications. Results: Nonspecific signal measured in drug-naive samples used to determine assay cut points was uniformly low and close to the screening cut point. Confirmatory assays performed on incurred study samples with nonspecific responses significantly above the level observed during cut point determination resulted in incorrect ADA classifications. Conclusion: Intensity of confirmatory response should be proportional to the screening response and therefore, to ensure accurate ADA classifications, the confirmatory responses cannot be considered as independent but need to be evaluated in relation to the screening responses.

Mitotic motor CENP-E cooperates with PRC1 in temporal control of central spindle assembly.

Error-free cell division depends on the accurate assembly of the spindle midzone from dynamic spindle microtubules to ensure chromatid segregation during metaphase-anaphase transition. However, the mechanism underlying the key transition from the mitotic spindle to central spindle before anaphase onset remains elusive. Given the prevalence of chromosome instability phenotype in gastric tumorigenesis, we developed a strategy to model context-dependent cell division using a combination of light sheet microscope and 3D gastric organoids. Light sheet microscopic image analyses of 3D organoids showed that CENP-E inhibited cells undergoing aberrant metaphase-anaphase transition and exhibiting chromosome segregation errors during mitosis. High-resolution real-time imaging analyses of 2D cell culture revealed that CENP-E inhibited cells undergoing central spindle splitting and chromosome instability phenotype. Using biotinylated syntelin as an affinity matrix, we found that CENP-E forms a complex with PRC1 in mitotic cells. Chemical inhibition of CENP-E in metaphase by syntelin prevented accurate central spindle assembly by perturbing temporal assembly of PRC1 to the midzone. Thus, CENP-E-mediated PRC1 assembly to the central spindle constitutes a temporal switch to organize dynamic kinetochore microtubules into stable midzone arrays. These findings reveal a previously uncharacterized role of CENP-E in temporal control of central spindle assembly. Since CENP-E is absent from yeast, we reasoned that metazoans evolved an elaborate central spindle organization machinery to ensure accurate sister chromatid segregation during anaphase and cytokinesis.

Neutrophil extracellular traps induced by IL-8 aggravate atherosclerosis via activation NF-κB signaling in macrophages.

Here, we sought to explore the underlying role of interleukin (IL)-8 in neutrophil extracellular traps (NETs) formation during atherosclerosis (AS). The concentration of pro-inflammatory cytokines IL-8, IL-6 and IL-1ß was determined by enzyme-linked immunosorbent assay (ELISA). NETs formation was evaluated by immunofluorescence and myeloperoxidase (MPO)-DNA complex ELISA. The mRNA levels of IL-8 and Toll-like receptor 9 (TLR9) were measured by quantitative real-time PCR (qRT-PCR). The phosphorylation levels of NF-κB p65 were detected by western blotting. The hematoxylin and eosin (H&E) staining of atherosclerotic lesion areas was performed in ApoE-deficiency mice. Results showed that patients with AS showed higher serum levels of IL-8, a pro-inflammatory cytokine and NETs. IL-8 interacted with its receptor CXC chemokine receptor 2 (CXCR2) on neutrophils, leading to the formation of NETs via Src and extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinases (MAPK) signaling to aggravate AS progression in vivo. PMA-induced NETosis directly upregulated the TLR9/NF-κB pathway in macrophages and subsequently initiated the release of IL-8. Our data reveal a neutrophil-macrophage interaction in AS progression, and indicate that NETs represent as a novel therapeutic target in treatment of AS and other cardiovascular diseases (CVD).

Potential Regulation Mechanisms of P-gp in the Blood-Brain Barrier in Hypoxia.

The blood-brain barrier (BBB) is a barrier of the central nervous system (CNS), which can restrict the free exchange of substances, such as toxins and drugs, between cerebral interstitial fluid and blood, keeping the relative physiological stabilization. The brain capillary endothelial cells, one of the structures of the BBB, have a variety of ATP-binding cassette transporters (ABC transporters), among which the most widely investigated is Pglycoprotein (P-gp) that can efflux numerous substances out of the brain. The expression and activity of P-gp are regulated by various signal pathways, including tumor necrosis factor-α (TNF-α)/protein kinase C-ß (PKC- ß)/sphingosine-1-phosphate receptor 1 (S1P), vascular endothelial growth factor (VEGF)/Src kinase, etc. However, it remains unclear how hypoxic signaling pathways regulate the expression and activity of P-gp in brain microvascular endothelial cells. According to previous research, hypoxia affects the expression and activity of the transporter. If the transporter is up-regulated, some drugs enter the brain's endothelial cells and are pumped back into the blood by transporters such as P-gp before they enter the brain tissue, consequently influencing the drug delivery in CNS; if the transporter is down-regulated, the centrally toxic drug would enter the brain tissue and cause serious adverse reactions. Therefore, studying the mechanism of hypoxia-regulating P-gp can provide an important reference for the treatment of CNS diseases with a hypoxia/reoxygenation (H/R) component. This article summarized the mechanism of regulation of P-gp in BBB in normoxia and explored that of hypoxia.