Glyphosate and tetrabromobisphenol A (TBBPA) are pollutants that pose a serious threat to the ecological safety of aquatic environments. However, there has been no report on the effects of combined exposure on the toxicity of carp fish gills in water. Therefore, we constructed a model of carp gill tissue and the carp epithelioma cells (EPC) cells exposed to glyphosate and/or TBBPA in vitro and in vivo, established a control group, a glyphosate group, a TBBPA group, and a glyphosate + TBBPA group, and added PI3K/AKT pathway activator musk ketone in vitro to verify the relationship between toxins and pathways. qRT-PCR and western blotting methods were used to detect the expression of oxidative stress-related indicators (CAT, GSH-Px, T-AOC, H2O2) and related genes. In vitro and in vivo results showed that glyphosate and/or TBBPA exposure resulted in overproduction of ROS, decreased activity of CAT, GSH-Px, T-AOC, and increased H2O2 content. Glyphosate and/or TBBPA exposure inhibited the PI3K/AKT/mTOR signaling pathway, further resulting in increased autophagy related genes LC3, ATG-5, Beclin-1, and decreased p62 expression. Inflammation related genes TNF-alpha, IL-1 beta, IL-6, IL-18 increased. And it was more significant when exposed in combination than when exposed alone. The addition of PI3K/AKT signaling pathway activator musk ketone in vitro can significantly alleviate the changes of autophagy and inflammation-related indicators. In summary, glyphosate and/or TBBPA induce oxidative stress by promoting gill autophagy and inflammation via the PI3K/AKT/mTOR pathway.

Nonlinear data visualization using t-distributed stochastic neighbor embedding (t-SNE) enables the representation of complex single-cell transcriptomic landscapes in two or three dimensions to depict biological populations accurately. However, t-SNE often fails to account for uncertainties in the original dataset, leading to misleading visualizations where cell subsets with noise appear indistinguishable. To address these challenges, we introduce uncertainty-aware t-SNE (Ut-SNE), a noise-defending visualization tool tailored for uncertain single-cell RNA-seq data. By creating a probabilistic representation for each sample, Our Ut-SNE accurately incorporates noise about transcriptomic variability into the visual interpretation of single-cell RNA sequencing data, revealing significant uncertainties in transcriptomic variability. Through various examples, we showcase the practical value of Ut-SNE and underscore the significance of incorporating uncertainty awareness into data visualization practices. This versatile uncertainty-aware visualization tool can be easily adapted to other scientific domains beyond single-cell RNA sequencing, making them valuable resources for high-dimensional data analysis.

Introduction Environmental and occupational exposure to cadmium (Cd) has been shown to cause acute kidney injury (AKI). Previous studies have demonstrated that autophagy inhibition and lysosomal dysfunction are important mechanisms of Cd-induced AKI. Objectives Transcription factor EB (TFEB) is a critical transcription regulator that modulates autophagy-lysosome function, but its role in Cd-induced AKI is yet to be elucidated. Thus, in vivo and in vitro studies were conducted to clarify this issue. Methods and results Data firstly showed that reduced TFEB expression and nuclear translocation were evident in Cd-induced AKI models, accompanied by autophagy-lysosome dysfunction. Pharmacological and genetic activation of TFEB improved Cd-induced AKI via alleviating autophagy inhibition and lysosomal dysfunction, whereas Tfeb knockdown further aggravated this phenomenon, suggesting the key role of TFEB in Cd-induced AKI by regulating autophagy. Mechanistically, Cd activated mechanistic target of rapamycin complex 1 (mTORC1) to enhance TFEB phosphorylation and thereby inhibiting TFEB nuclear translocation. Cd also activated chromosome region maintenance 1 (CRM1) to promote TFEB nuclear export. Meanwhile, Cd activated general control non-repressed protein 5 (GCN5) to enhance nuclear TFEB acetylation, resulting in the decreased TFEB transcriptional activity. Moreover, inhibition of CRM1 or GCN5 alleviated Cd-induced AKI by enhancing TFEB activity, respectively. Conclusion In summary, these findings reveal that TFEB phosphorylation, nuclear export and acetylation independently suppress TFEB activity to cause Cd-induced AKI via regulating autophagy-lysosome function, suggesting that TFEB activation might be a promising treatment strategy for Cd-induced AKI.

SHP2 plays a critical role in modulating tumor growth and PD-1-related signaling pathway, thereby serving as an attractive antitumor target. To date, no antitumor drugs targeting SHP2 have been approved, and hence, the search of SHP2 inhibitors with new chemical scaffolds is urgently needed. Herein, we developed a novel SHP2 allosteric inhibitor SDUY038 with a furanyl amide scaffold, demonstrating potent binding affinity (K D = 0.29 μM), enzymatic activity (IC50 = 1.2 μM) and similar binding interactions to SHP099. At the cellular level, SDUY038 exhibited pan-antitumor activity (IC50 = 7–24 μM) by suppressing pERK expression. Furthermore, SDUY038 significantly inhibited tumor growth in both xenograft and organoid models. Additionally, SDUY038 displayed acceptable bioavailability (F = 14%) and half-life time (t 1/2 = 3.95 h). Conclusively, this study introduces the furanyl amide scaffold as a novel class of SHP2 allosteric inhibitors, offering promising lead compounds for further development of new antitumor therapies targeting SHP2.

Due to the sequential sample arrival, changing experiment conditions, and evolution of knowledge, the demand to continually visualize evolving structures of sequential and diverse single-cell RNA-sequencing (scRNA-seq) data becomes indispensable. However, as one of the state-of-the-art visualization and analysis methods for scRNA-seq, t-distributed stochastic neighbor embedding (t-SNE) merely visualizes static scRNA-seq data offline and fails to meet the demand well. To address these challenges, we introduce online t-SNE to seamlessly integrate sequential scRNA-seq data. Online t-SNE achieves this by leveraging the embedding space of old samples, exploring the embedding space of new samples, and aligning the two embedding spaces on the fly. Consequently, online t-SNE dramatically enables the continual discovery of new structures and high-quality visualization of new scRNA-seq data without retraining from scratch. We showcase the formidable visualization capabilities of online t-SNE across diverse sequential scRNA-seq datasets.

Phage therapy has shown great promise for the treatment of multidrug-resistant bacterial infections. However, the lack of a thorough and organized understanding of phage-body interactions has limited its clinical application. Here, we administered different purified phages (Salmonella phage SE_SZW1, Acinetobacter phage AB_SZ6, and Pseudomonas phage PA_LZ7) intravenously to healthy animals (rats and monkeys) to evaluate the phage-induced host responses and phage pharmacokinetics with different intravenous (IV) doses in healthy animals. The plasma and the organs were sampled after different IV doses to determine the phage biodistribution, phage-induced cytokines, and antibodies. The potential side effects of phages on animals were assessed. A non-compartment model revealed that the plasma phage titer gradually decreased over time following a single dose. Repeated doses resulted in a 2-3 Log10 decline of the plasma phage titer at 5 min compared to the first dose, regardless of the type of phage administered in rats. Host innate immune responses were activated including splenic enlargement following repeated doses. Phage-specific neutralization antibodies in animals receiving phages were detected. Similar results were obtained from monkeys. In conclusion, the mammalian bodies were well-tolerant to the administered phages. The animal responses to the phages and the phage biodistribution profiles could have a significant impact on the efficacy of phage therapy.IMPORTANCEPhage therapy has demonstrated potential in addressing multidrug-resistant bacterial infections. However, an insufficient understanding of phage-host interactions has impeded its broader clinical application. In our study, specific phages were administered intravenously (IV) to both rats and monkeys to elucidate phage-host interactions and evaluate phage pharmacokinetics (PK). Results revealed that with successive IV administrations, there was a decrease in plasma phage concentrations. Concurrently, these administrations elicited both innate and adaptive immune responses in the subjects. Notably, the observed immune responses and PK profiles exhibited variation contingent upon the phage type and the mammalian host. Despite these variations, the tested mammals exhibited a favorable tolerance to the IV-administered phages. This underscores the significance of comprehending these interactions for the optimization of phage therapy outcomes.

Oxidized low density lipoprotein (oxLDL)-induced endothelial oxidative damage promotes the development of atherosclerosis. Caveolae play an essential role in maintaining the survival and function of vascular endothelial cell (VEC). It is reported that the long coiled-coil protein NECC2 is localized in caveolae and is associated with neural cell differentiation and adipocyte formation, but its role in VECs needs to be clarified. Our results showed NECC2 expression increased in the endothelium of plaque-loaded aortas and oxLDL-treated HUVECs. Down-regulation of NECC2 by NECC2 siRNA or compound YF-307 significantly inhibited oxLDL-induced VEC apoptosis and the adhesion factors expression. Remarkably, inhibition of NECC2 expression in the endothelium of apoE-/- mice by adeno-associated virus (AAV)-carrying NECC2 shRNA or compound YF-307 alleviated endothelium injury and restricted atherosclerosis development. The immunoprecipitation results confirmed that NECC2 interacted with Tyk2 and caveolin-1(Cav-1) in VECs, and NECC2 further promoted the phosphorylation of Cav-1 at Tyr14 b y activating Tyk2 phosphorylation. On the other hand, inhibiting NECC2 levels suppressed oxLDL-induced phosphorylation of Cav-1, uptake of oxLDL by VECs, accumulation of intracellular reactive oxygen species and activation of NF-κB. Our findings suggest that NECC2 may contribute to oxLDL-induced VEC injury and atherosclerosis via modulating Cav-1 phosphorylation through Tyk2. This work provides a new concept and drug target for treating atherosclerosis.

Ferroptosis is a nonapoptotic, iron-catalyzed form of regulated cell death. It has been shown that high glucose (HG) could induce ferroptosis in vascular endothelial cells (VECs), consequently contributing to the development of various diseases. This study synthesized and evaluated a series of novel ferrostatin-1 (Fer-1) derivatives fused with a benzohydrazide moiety to prevent HG-induced VEC ferroptosis. Several promising compounds showed similar or improved inhibitory effects compared to positive control Fer-1. The most effective candidate 12 exhibited better protection against erastin-induced ferroptosis and high glucose-induced ferroptosis in VECs. Mechanistic studies revealed that compound 12 prevented mitochondrial damage, reduced intracellular ROS accumulation, upregulated the expression of GPX4, and decreased the amounts of ferrous ion, LPO and MDA in VECs. However, compound 12 still exhibited undesirable microsomal stability like Fer-1, suggesting the need for further optimization. Overall, the present findings highlight ferroptosis inhibitor 12 as a potential lead compound for treating ferroptosis-associated vascular diseases. This work designed and synthesized novel ferrostatin analogs with a benzohydrazide moiety, and identified compound 12 as a promising lead for preventing HG-induced VEC ferroptosis.

BACKGROUND:Currently, there is increasing evidence from clinic, epidemiology, as well as neuroimaging, demonstrating neuropsychiatric abnormalities in COVID-19, however, whether there were associations between brain changes caused by COVID-19 and genetic susceptibility of psychiatric disorders was still unknown.METHODS:In this study, we performed a meta-analysis to investigate these associations by combing single-cell RNA sequencing datasets of brain tissues of COVID-19 and genome-wide association study summary statistics of psychiatric disorders.RESULTS:The analysis demonstrated that among ten psychiatric disorders, gene expression perturbations implicated by COVID-19 in excitatory neurons of choroid plexus were significantly associated with schizophrenia.CONCLUSIONS:Our analysis might provide insights for the underlying mechanism of the psychiatric consequence of COVID-19.

GW117 is new melatonergic antidepressant being developed to show better antidepressant action than agomelatine. The purpose of this study was to evaluate the toxicity and to determine potential target organs after oral (gavage) administration of the test article GW117 for 28 days and to assess the reversibility after a 4-week recovery phase in beagle dogs. Toxicokinetics was also evaluated. Four groups were designed in this study, including the vehicle control group and the GW117 50, 150 and 500 mg/kg/day groups, with 5 dogs/sex/group. Body weight, hematology, clinical chemistry, gross necropsy, organ weight, histopathology, and other indicators were examined. Results showed that animals dosed at >= 150 mg/kg/day showed gastrointestinal reactions (watery feces and dark green/red brown feces), with a dose-response relationship in the incidence and severity grade. Female dogs at 500 mg/kg/day had an increase in organ weight and ratios of the liver at the end of the dosing phase. Histopathology examination showed that some animals at 500 mg/kg/day, especially female animals, had minimal centrilobular hepatocyte hypertrophy in the liver, which reversed after 28-day recovery. With the exception of the above, no GW117-related abnormality was noted. Meanwhile, there were no sexual differences in drug exposure and accumulation after the first and last dosing. The no observed adverse effect dose level (NOAEL) was 150 mg/kg/day, under which mean C-max and AUC(0 -> t) were 583.5 and 2767.0 ng/ml*h for females and 663.2 and 4046.3 ng/ml*h for males on Day 28.

Objectives Phage therapy has shown a great promise for the treatment of multidrug-resistant bacterial infections. However, the lack of a thorough and organized understanding of phage-body interactions has limited its clinical application. Methods Here, we administered different purified phages ( Salmonella phage SE_SZW1, Acinetobacter phage AB_SZ6, and Pseudomonas phage PA_LZ7) intravenously to healthy animals (rats and monkeys) to evaluate the phage-induced host responses and phage pharmacokinetics (PK) with different intravenous (IV) doses in healthy animals. The plasma and the organs were sampled after different IV doses to determine the phage biodistribution, the phage-induced cytokines, and antibodies. The potential side effects of phages on animals were assessed. Results A non-compartment model revealed that the plasma phage titer gradually decreased over time following a single dose. Repeated doses caused that the plasma phage titer at 5 minutes dropped 2-3 Log 10 compared to the first dose regardless of phage types in rats. Host innate immune responses were activated including the upregulated expression (>10-fold) of TNF- α and splenic enlargement following repeated doses. Phage-specific neutralization antibodies in animals receiving phages were detected. Similar results were obtained from monkeys. Conclusions The mammalian bodies were well-tolerant to the administered phages. The animal responses to the phages and the phage biodistribution profiles could have a significant impact on the efficacy of phage therapy.

ABSTRACT Phage therapy has become one of the most promising alternatives to antibiotics as the surge of multi-drug-resistant (MDR) bacteria severely threatens global public health. The lack of a thorough understanding of pharmacokinetics (PK) and the safety study of phage has limited its clinical application. In this study, we aimed to evaluate the PK and safety of a lytic phage PA_LZ7 that infects MDR Pseudomonas aeruginosa using intravenous (IV) route in an uninfected immune-competent mice model. The active phage of PA_LZ7 exhibited an exponential decay in plasma at 2*108 (low dosage), 2*1010 [medium dosage (MD)], and 2*1011 PFU/kg [high dosage (HD)] after IV administration, and the phage titer dropped to ~4 log10 PFU/mL over 24 h for all dosages. Phages primarily accumulated in the spleen at 1 h post-administration and gradually decreased in all dosages. The active phage titer in other organs decreased globally within 72 h as well. Plasma cytokines IL-6, IL-10, KC/GRO, and tumor necrosis factor-alpha sharply increased 1 h post-administration in mice of MD and HD groups and dropped to a normal range within 24 h. No obvious toxic effect was observed, except for the increased spleen weight and relative spleen weight in the MD and HD group and the slight lymphocytosis in the white pulp of the spleen in the HD group. In conclusion, PA_LZ7 possesses good PK properties and shows no obvious toxicity; it could be considered as a potent therapeutic candidate for the treatment of MDR P. aeruginosa infections in future phage therapy. IMPORTANCE Phage therapy is gaining traction as an alternative to antibiotics due to the rise of multi-drug-resistant (MDR) bacteria. This study assessed the pharmacokinetics and safety of PA_LZ7, a phage targeting MDR Pseudomonas aeruginosa, in mice. After intravenous administration, the phage showed an exponential decay in plasma and its concentration dropped significantly within 24 h for all dosage groups. Although there was a temporary increase in certain plasma cytokines and spleen weight at higher dosages, no significant toxicity was observed. Therefore, PA_LZ7 shows potential as an effective and safe candidate for future phage therapy against MDR P. aeruginosa infections.

Glyphosate (GLY), a ubiquitous environmental pollutant, can result in gut microbiota dysbiosis intimately involving various diseases. The latest research has shown an association between gut microbiota alteration and defective spermatogenesis. Here, we aimed to investigate whether GLY-induced gut microbiota dysbiosis contributed to male reproductive toxicity. Data showed that GLY-exposed rats exhibited male reproductive dysfunction, evidenced by impaired testis architectural structure, reduced sperm motility, together with increased sperm malformation ratio. 16S rDNA sequencing analysis indicated that GLY exposure altered the composition of gut commensal microbiota, of which the relative abundance of Bacteroidetes and Firmicutes phyla was significantly changed. Unexpectedly, the increased abundance of Prevotella_1 and Bacteroides genera was negatively correlated with sperm quality. Mechanistically, the pathological changes in GLY-exposed testis were accompanied by the increased interleukin (IL)-17A production, probably due to gut microbes-derived Th17 cell migration. Furthermore, activation of IL-17A signaling triggered testicular oxidative damage. Taken together, these findings uncover an underlying mechanistic scenario that gut microbiota dysbiosis-driven local IL-17A production is one reason responsible for male reproductive toxicity induced by GLY, which provides new insights into the male reproductive toxicity of GLY in mammals.

Glyphosate (GLY), an active ingredient of the most commonly used herbicide, when in crops and feed, is deleterious to male reproductive health. Trehalose (Tre), a naturally non-reducing disaccharide, is shown to counteract the adverse stresses due to its antioxidation effect. Thus, this study was designed to investigate whether Tre can improve GLY-induced testicular damage via suppressing oxidative stress. 60 healthy Hy-Line Brown breeder roosters were utilized to assess the protective effects of Tre supplementation against testicular oxidative damage caused by GLY. Data showed that Tre administration significantly alleviated GLY- induced reduction in testis weight, decreased GLY level in the testis tissues, and alleviated GLY-caused testicular pathological damage. Concurrently, GLY treatment significantly elevated serum malondialdehyde (MDA) and testicular reactive oxygen species (ROS) levels, decreased serum total anti-oxidation capacity (T-AOC), catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels, which were all notably reversed by Tre administration. Moreover, GLY- inhibited nuclear factor erythroid 2-related factor 2 (Nrf2) pathway in rooster testis, a master regulator of oxidative stress, was markedly recovered by Tre administration. In summary, these findings demonstrated that Tre can prevent GLY-induced testicular damage in roosters by ameliorating oxidative stress.

目的 基于遥测技术建立适用于监测清醒自由活动状态下巴马小型猪心电、血压、体温等生理指标的动物模型,为其在安全药理学研究中的应用提供支持.方法 8只巴马小型猪(雌雄各半)经手术植入植入子,手术后恢复3～4周,经体格检查确认已完全恢复后,采用EMKA遥测系统连续监测至少24 h心电图、血压等生理指标,包括心率、PR间期、QRS间期、QT间期、校正QT(QTcv)间期、体温、收缩压(SBP)、舒张压(DBP)、平均动脉压(MBP),并于次周重复监测1次.对巴马小型猪的24h指标变化进行分析,并与Beagle犬和食蟹猴数据进行种属间比对分析.结果 巴马小型猪的心电数据和血压数据昼夜节律均不明显;体温呈现明显的昼夜节律,白天体温高,夜间体温低;动物的心率、血压和体温均受外界影响比较大,与动物的活动呈现明显的相关性;与Beagle犬和食蟹猴比较,巴马小型猪的心率和人类最为相似,SBP仅稍高于人类.结论 通过体内植入植入子,成功建立了巴马小型猪植入式遥测动物模型;与Beagle犬和食蟹猴比较,巴马小型猪更适合用于药物对心血管系统影响的评价.

As a powerful tool for the discovery of peptide compounds, high-throughput library screening technology can also perform multitarget screening with high chemical diversity as a condition, which accelerates the process of exploring high-affinity peptide molecules. These techniques can be generally divided into in vitro and in vivo screening techniques, both of which play important roles in peptide drug discovery. Compared with linear peptides, cyclic peptides have been proven to have good bioavailability, membrane permeability, and stability and longer half-lives and have gradually become a topic of high interest in peptide drug research. High-throughput library screening technology plays an important role in the development of cyclic peptide drugs. Based on published studies, this paper reviews the high-throughput library screening technology used in the development of cyclic peptide drugs.

Oxidized low-density lipoprotein (oxLDL)-induced endothelium injury promotes the development of atherosclerosis. It has been reported that homoplantaginin, a flavonoid glycoside from the traditional Chinese medicine Salvia plebeia R. Br., protected vascular endothelial cells by inhibiting inflammation. However, it is undetermined whether homoplantaginin affects atherosclerosis. In this study, we evaluated the effect of homoplantaginin and its derivative dihydrohomoplantagin on oxLDL-induced endothelial cell injury and atherosclerosis in apoE-/- mice. Our results showedthat both dihydrohomoplantagin and homoplantaginin inhibited apoptosis and the increased level of ICAM-1 and VCAM-1 in oxLDL-stimulated HUVECs and the plaque endothelium of apoE-/- mice. Additionally, both of them restricted atherosclerosis development of apoE-/- mice. Mechanistic studies showed that oxLDL-induced the increase in ROS production, phosphorylation of ERK and nuclear translocation of NF-κB in HUVECs was significantly inhibited by the compounds. Meanwhile, these two compounds promoted Nrf2 nuclear translocation and increased the anti-oxidation downstream HO-1 protein level in HUVECs and plaque endothelium. Notably, knockdown of Nrf2 by siRNA abolished the cell protective effects of compounds and antagonized the inhibition effects of them on ROS production and NF-κB activation in oxLDL-stimulated HUVECs. Collectively, dihydrohomoplantagin and homoplantaginin protected VECs by activating Nrf2 and thus inhibited atherosclerosis in apoE-/- mice.

Glyphosate (GLY) frequently detected in various water bodies has imposed a serious risk on fish. Head kidney of fish is an important defense organ, playing a vital part in antagonizing exogenous hazardous matter. The objective of this study was to characterize toxic mechanisms of GLY in head kidney of common carp based on transcriptome profiling. After 45-days exposure of GLY at environmentally relevant concentrations, juvenile common carp were used as experimental subjects to analyze how the head kidney responded to GLY. The transcriptome profiling identified 1381 different expressed genes (DEGs) between the control and exposure groups (5 and 50 mg/L). Functional analysis of DEGs substantiated over-representative pathways mainly involving cellular stress responses, cell proliferation and turnover, apoptosis, lipid metabolism, and innate immune processes in both treated groups compared with the control group. Predicted network of gene regulation indicated that GLY-induced tp53 played a vital role in linking a battery of signals. Furthermore, the expression of 10 candidate genes by qRT-PCR aligned with transcriptional profiling. In addition, western blotting analysis confirmed that GLY-induced apoptosis and cellular proliferation were closely involved in activating MAKP signaling pathway and lipid metabolism pathway in both treated groups. Collectively, these data demonstrate that head kidney of juvenile common carp mainly leverages upregulation of genes related to cell proliferation and turnover, apoptosis, and lipid metabolism to combat sub-chronic exposure of GLY. This study casts new understanding into the risk of GLY in aquatic animals.