Search

To study the biocompatibility of nanohydroxyapatite (nmHA)-SiO 2 fiber material and its efficacy in guided bone regeneration. ① The cytotoxicity of the nmHA-SiO 2 fiber material to MC3T3-E1 cells was determined by CCK-8 assay. The adhesion of cells on the surface of the material was observed. ② Bone defects were prepared in the skull of three groups of New Zealand white rabbits. The following treatments were administered: implantation of nmHA-SiO 2 , implantation of Bio-Oss, and no treatment. The defects were then covered with nmHA-SiO 2 membrane or Hai'ao oral repair membrane. Animal samples were analyzed by gross observation, micro-computed tomography, hematoxylin-eosin staining and Masson staining. The data were statistically analyzed by multivariate analysis of variance to evaluate the repair of bone defects. ① The nmHA-SiO 2 fiber material has suitable biocompatibility. ② The nmHA-SiO 2 fiber material performed more effectively as a barrier membrane than other bone substitute materials in GBR model rabbits.

The simultaneous application of photothermal therapy (PTT) and photodynamic therapy (PDT) offers substantial advantages in cancer treatment. However, their synergistic anticancer efficacy is often limited by tumor hypoxia, and thermotolerance induced by high expression of heat shock proteins (HSP). Fortunately, hydrogen sulfide (H 2 S), known for its direct cytotoxic effect on tumor cells, has been recognized for its ability to enhance PTT and PDT. The effectiveness of H 2 S in these therapies is challenged by its low loading efficiency, poor stability, and short diffusion distance. To address these issues, a nanoscale emulsion drop template created through the salting-out effect is employed to construct a robust H 2 S delivery system. Polydopamine (PDA), chosen for its interfacial polymerization tendency and excellent photothermal conversion rate, is utilized as a carrier for the H 2 S donor (ADT) and Zinc phthalocyanine (ZnPc) to fabricate a novel nanomedicine termed APZ NPs. The temperature-responsive APZ NPs are designed to release H 2 S during the PTT process. Elevated H 2 S levels promoted vasodilation, thereby enhancing the enhanced permeability and retention effect (EPR) of APZ NPs within solid tumors. This strategy effectively alleviated tumor hypoxia by disrupting the mitochondrial respiratory chain and mitigated tumor cell heat tolerance by inhibiting HSP expression., (© 2024 Wiley‐VCH GmbH.)

Quorum sensing (QS)-based manipulations emerge as a promising solution for biofilm reactors to overcome challenges from inefficient biofilm formation and lengthy start-ups. However, the ecological mechanisms underlying how QS regulates microbial behaviors and community assembly remain elusive. Herein, by introducing different levels of N-acyl-homoserine lactones, we manipulated the strength of QS during the start-up of moving bed biofilm reactors and compared the dynamics of bacterial communities. We found that enhanced QS elevated the fitness of fast-growing bacteria with high ribosomal RNA operon (rrn) copy numbers in their genomes in both the sludge and biofilm communities. This led to notably increased extracellular substance production, as evidenced by strong positive correlations between community-level rrn copy numbers and extracellular proteins and polysaccharides (Pearson's r = 0.529-0.830, P < 0.001). Network analyses demonstrated that enhanced QS significantly promoted the ecological interactions among taxa, particularly cooperative interactions. Bacterial taxa with higher network degrees were more strongly correlated with extracellular substances, suggesting their crucial roles as public goods in regulating bacterial interactions and shaping network structures. However, the assembly of more cooperative communities in QS-enhanced reactors came at the cost of decreased network stability and modularity. Null model and dissimilarity-overlap curve analysis revealed that enhanced QS strengthened stochastic processes in community assembly and rendered the universal population dynamics more convergent. Additionally, these shaping effects were consistent for both the sludge and biofilm communities, underpinning the planktonic-to-biofilm transition. This work highlights that QS manipulations efficiently drive community assembly and confer specialized functional traits to communities by recruiting taxa with specific life strategies and regulating interspecific interactions. These ecological insights deepen our understanding of the rules governing microbial societies and provide guidance for managing engineering ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Despite exhibiting potent anticancer activity, the strong hemolytic properties of melittin (MEL) significantly restrict its delivery efficiency and clinical applications. To address this issue, we have devised a strategy wherein homologous dopamine (DA), an essential component of bee venom, is harnessed as a vehicle for the synthesis of MEL-polydopamine (PDA) nanoparticles (MP NPs). The ingenious approach lies in the fact that MEL is a basic polypeptide, and the polymerization of DA is also conducted under alkaline conditions, indicating the distinctive advantages of PDA in MEL encapsulation. Furthermore, MP NPs are modified with folic acid to fabricate tumor-targeted nanomedicine (MPF NPs). MPF NPs can ameliorate the hemolysis of MEL in drug delivery and undergo degradation triggered by high levels of reactive oxygen species (ROS) within solid tumors, thereby facilitating MEL release and subsequent restoration of anticancer activity. After cellular uptake, MPF NPs induce cell apoptosis through the PI3K/Akt-mediated p53 signaling pathway. The tumor growth inhibitory rate of MPF NPs in FA receptor-positive 4T1 and CT26 xenograft mice reached 78.04% and 81.66%, which was significantly higher compared to that in FA receptor-negative HepG2 xenograft mice (45.79%). Homologous vehicles provide a new perspective for nanomedicine design.

Objective: Since diabetic patients with coronary microvascular dysfunction (CMD) exhibit high cardiac mortality and women have higher prevalence of non-obstructive coronary artery disease than men, we tried to expand the limited understanding about the etiology and the sex difference of diabetic CMD., Approach and Results: Accumulated methylglyoxal (MGO) due to diabetes promotes vascular damage and it was used for mimicking diabetic status. Flow cytometry analysis and isometric tension measurement were performed to evaluate coronary artery endothelial injury. MGO induced apoptosis of coronary endothelial cells, accompanied by downregulation of androgen receptor (AR). Lentivirus-mediated stable expression of AR in coronary endothelial cells increased anti-apoptotic Bcl-2 expression and attenuated MGO-induced cell apoptosis. cPLA 2 activation was the downstream of AR downregulation by MGO treatment. Moreover, MGO also activated cPLA 2 rapidly to impair endothelium-dependent vasodilation of coronary arteries from mice. Reactive oxygen species (ROS) overproduction was demonstrated to account for MGO-mediated cPLA 2 activation and endothelial dysfunction. Importantly, AR blockade increased endothelial ROS production whereas AR activation protected coronary artery endothelial vasodilatory function from the MGO-induced injury. Although galectin-3 upregulation was confirmed by siRNA knockdown in endothelial cells not to participate in MGO-induced endothelial apoptosis, pharmacological inhibitor of galectin-3 further enhanced MGO-triggered ROS generation and coronary artery endothelial impairment., Conclusions: Our data proposed the AR downregulation-ROS overproduction-cPLA 2 activation pathway as one of the mechanisms underlying diabetic CMD and postulated a possible reason for the sex difference of CMD-related angina. Meanwhile, MGO-induced galectin-3 activation played a compensatory role against coronary endothelial dysfunction., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Period circadian regulator 3 (PER3) functions as a tumor suppressor in various cancers. However, the role of PER3 in multiple myeloma (MM) has not been reported yet. Through this study, we aimed to investigate the potential role of PER3 in MM and the underlying mechanisms. RT-qPCR and western blotting were used to determine the mRNA and protein expression levels of PER3. Glyoxylate reductase 1 homolog (GLYR1) was predicted to be a transcription factor of PER3. The binding sites of GLYR1 on the promoter region of PER3 were analyzed using UCSC and confirmed using luciferase and chromatin immunoprecipitation assays. Viability, apoptosis, and metathesis were determined using CCK-8, colony formation, TUNEL, and transwell assays. We found that PER3 expression decreased in MM. Low PER3 levels may predict poor survival rates; PER3 overexpression suppresses the viability and migration of MM cells and promotes apoptosis. Moreover, GLYR1 transcriptionally activates PER3, and the knockdown of PER3 alleviates the effects of GLYR1 and induces its malignant behavior in MM cells. To conclude, GLYR1 upregulates PER3 and suppresses the aggressive behavior of MM cells, suggesting that GLYR1/PER3 signaling may be a potential therapeutic target for MM., Competing Interests: Declaration of competing interest The authors declared that they have none conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The ecological impact of microplastics (MPs) in coastal environments has been widely studied. However, the influence of small microplastics in the actual environment is often overlooked due to measurement challenges. In this study, Hangzhou Bay (HZB), China, was selected as our study area. High-throughput metagenomic sequencing and micro-Raman spectrometry were employed to analyze the microbial communities and microplastics of coastal sediment samples, respectively. We aimed to explore the ecological impact of MPs with small sizes (≤ 100 μm) in real coastal sediment environments. Our results revealed that as microplastic size decreased, the environmental behavior of MPs underwent alterations. In the coastal sediments, no significant correlations were observed between the detected MPs and the whole microbial communities, but small MPs posed potential hazards to eukaryotic communities. Moreover, these small MPs were more prone to microbial degradation and significantly affected carbon metabolism in the habitat. This study is the first to reveal the comprehensive impact of small MPs on microbial communities in a real coastal sediment environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Plant leaf senescence, caused by multiple internal and environmental factors, has an important impact on agricultural production. The lectin receptor-like kinase (LecRLK) family members participate in plant development and responses to biotic and abiotic stresses, but their roles in regulating leaf senescence remain elusive. Here, we identify and characterize a rice premature withered leaf 1 (pwl1) mutant, which exhibits premature leaf senescence throughout the plant life cycle. The pwl1 mutant displayed withered and whitish leaf tips, decreased chlorophyll content, and accelerated chloroplast degradation. Map-based cloning revealed an amino acid substitution (Gly412Arg) in LOC&#95;Os03g62180 (PWL1) was responsible for the phenotypes of pwl1. The expression of PWL1 was detected in all tissues, but predominantly in tillering and mature leaves. PWL1 encodes a G-type LecRLK with active kinase and autophosphorylation activities. PWL1 is localized to the plasma membrane and can self-associate, mainly mediated by the plasminogen-apple-nematode (PAN) domain. Substitution of the PAN domain significantly diminished the self-interaction of PWL1. Moreover, the pwl1 mutant showed enhanced reactive oxygen species (ROS) accumulation, cell death, and severe DNA fragmentation. RNA sequencing analysis revealed that PWL1 was involved in the regulation of multiple biological processes, like carbon metabolism, ribosome, and peroxisome pathways. Meanwhile, interfering of biological processes induced by the PWL1 mutation also enhanced heat sensitivity and resistance to bacterial blight and bacterial leaf streak with excessive accumulation of ROS and impaired chloroplast development in rice. Natural variation analysis indicated more variations in indica varieties, and the vast majority of japonica varieties harbour the PWL1 Hap1 allele. Together, our results suggest that PWL1, a member of LecRLKs, exerts multiple roles in regulating plant growth and development, heat-tolerance, and resistance to bacterial pathogens., (© 2023 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

Wastewater treatment plants (WWTPs) effluent often contains a significant amount of residual organic pollutants and nutrients, causing disturbance to the coastal effluent receiving areas (ERA). Microbial communities in coastal ERA sediments may benefit from the coexistence of organic pollutants and nutrients, promoting the emergence of versatile taxa that are capable of eliminating these substances simultaneously. However, the identification and exploration of versatile taxa in natural environments under anthropogenic disturbances remain largely uncharted territory. In this study, we specifically focused on the versatile taxa coupled by the degradation of aromatic compounds (ACs) and denitrification, using Hangzhou Bay in China as our study area. We explored how WWTPs effluent disturbance would affect the versatile taxa, and particularly examined the role of disturbance intensity in shaping their composition. Intriguingly, we found that versatile taxa were mainly derived from denitrifiers like Pseudomonas, suggesting the fulfilled potential of denitrifiers regarding ACs degradation. We also discovered that moderate disturbance stimulated the diversity of versatile taxa, resulting in strengthened functional redundancy. Through correlation network analysis, we further demonstrated that moderate disturbance enhanced the community-level cooperation. Thus, moderate disturbance serves as a catalyst for versatile taxa to maintain community function, making them more resilient to effluent disturbances. Additionally, we identified COD and NO 3 - -N concentrations as significant environmental factors influencing the versatile taxa. Overall, our findings reveal the role of effluent disturbances in the promotion of versatile taxa, and highlight moderate disturbance can foster more robust versatile taxa that are better equipped to handle effluent disturbances., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

N6-methyladenosine (m 6 A) modification has been implicated in the progression of obesity and metabolic diseases. However, its impact on beige fat biology is not well understood. Here, via m 6 A-sequencing and RNA-sequencing, this work reports that upon beige adipocytes activation, glycolytic genes undergo major events of m 6 A modification and transcriptional activation. Genetic ablation of m 6 A writer Mettl3 in fat tissues reveals that Mettl3 deficiency in mature beige adipocytes leads to suppressed glycolytic capability and thermogenesis, as well as reduced preadipocytes proliferation via glycolytic product lactate. In addition, specific modulation of Mettl3 in beige fat via AAV delivery demonstrates consistently Mettl3's role in glucose metabolism, thermogenesis, and beige fat hyperplasia. Mechanistically, Mettl3 and m 6 A reader Igf2bp2 control mRNA stability of key glycolytic genes in beige adipocytes. Overall, these findings highlight the significance of m 6 A on fat biology and systemic energy homeostasis., (© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH.)

The biosensor is an instrument that converts the concentration of biomarkers into electrical signals for detection. Biosensing technology is non-invasive, lightweight, automated, and biocompatible in nature. These features have significantly advanced medical diagnosis, particularly in the diagnosis of mental disorder in recent years. The traditional method of diagnosing mental disorders is time-intensive, expensive, and subject to individual interpretation. It involves a combination of the clinical experience by the psychiatrist and the physical symptoms and self-reported scales provided by the patient. Biosensors on the other hand can objectively and continually detect disease states by monitoring abnormal data in biomarkers. Hence, this paper reviews the application of biosensors in the detection of mental diseases, and the diagnostic methods are divided into five sub-themes of biosensors based on vision, EEG signal, EOG signal, and multi-signal. A prospective application in clinical diagnosis is also discussed., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zheng, Liu, Lai, Wang, Xia, Sun and Zhang.)

The wastewater treatment plant (WWTP) effluent discharge affects the microorganisms in the receiving water bodies. Despite the ecological significance of microbial communities in pollutant degradation and element cycling, how the community diversity is affected by effluent remains obscure. Here, we compared the sediment bacterial communities exposed to different intensities of WWTP effluent discharge in Hangzhou Bay, China: i) a severely polluted area that receives effluent from an industrial WWTP, ii) a moderately polluted area that receives effluent from a municipal WWTP, and iii) less affected area that inner the bay. We found that the sediment bacterial diversity decreased dramatically with pollution levels of inorganic nutrients, heavy metals, and organic halogens. Microbial community assembly model analysis revealed increased environmental selection and decreased species migration rate in the severely polluted area, resulting in high phylogenetic clustering of the bacterial communities. The ecological networks were less complex in the two WWTP effluent receiving areas than in the inner bay area, as suggested by the smaller network size and lower modularity. Fewer negative network associations were detected in the severely (6.7%) and moderately (8.3%) polluted areas than in the less affected area (16.7%), indicating more collaborative inter-species behaviors are required under stressful environmental conditions. Overall, our results reveal the fundamental impacts of WWTP effluents on the ecological processes shaping coastal microbial communities and point to the potential adverse effects of diversity loss on ecosystem functions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Sodium humate (SH) is one of the derivatives humic substances, which can be utilized for heavy metal removal from water due to its containing plenty of functional groups. In this study, a double network hydrogel SH/polyacrylamide (SH/PAM) was synthesized by a simple free-radical polymerization and used for Cu 2+ and Pb 2+ removal from water. The adsorption process can be well described by Langmuir-Freundlich model, indicating that both physical and chemical adsorption were involved. X-ray photoelectron spectroscopy (XPS) characterization demonstrated that complexation was the main mechanism for the adsorption. Two-dimensional correlation analysis of FTIR (2D-FTIR-COS) results showed that the variation order of functional groups during Cu 2+ and Pb 2+ adsorption in the following order: COOH ≈ -CO > -OH > C-O and -COOH ≈ C-O > -CO > -OH, respectively. According to the density functional theory (DFT) calculation results, the O atom of SH in the COO - was the main adsorption site. Meanwhile, the adsorption energy of Pb 2+ was more negative than that of Cu 2+ and the orbital hybridization between O atom of SH and Pb 2+ was denser than that of Cu 2+ , which suggested that SH/PAM had a stronger combining capacity for Pb 2+ than Cu 2+ . Therefore, the adsorption capacity for Pb 2+ was larger than Cu 2+ . Moreover, the removal efficiencies are 30.2% for Al, 98.79% for Cu, 99.0% for Fe, 17.2% for Mn, 93.4% for Pb, and 62.4% for Zn in actual acid mine drainage using 6 g L -1 adsorbent. Collectively, this study not only provided a new adsorbent for heavy metal removal but also explicated the mechanism of heavy metal removal by SH from molecule and electron perspective, which is helpful for the application of SH in the environmental field., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Wastewater treatment plants (WWTPs) have been regarded as an important source of antibiotic resistance genes (ARGs) in environment, but out of municipal domestic WWTPs, few evidences show how environment is affected by industrial WWTPs. Here we chose Hangzhou Bay (HZB), China as our study area, where land-based municipal and industrial WWTPs discharged their effluent into the bay for decades. We adopted high-throughput metagenomic sequencing to examine the antibiotic resistome of the WWTP effluent and coastal sediment samples. And we proposed a conceptual framework for the assessment of antibiotic resistome risk, and a new bioinformatic pipeline for the evaluation of the potential horizontal gene transfer (HGT) frequency. Our results revealed that the diversity and abundance of ARGs in the WWTP's effluent were significantly higher than those in the sediment. Furthermore, the antibiotic resistome in the effluent-receiving area (ERA) showed significant difference from that in HZB. For the first time, we identified that industrial WWTP effluent boosted antibiotic resistome risk in coastal sediment. The crucial evidences included: 1) the proportion of ARGs derived from WWTP activated sludge (WA) was higher (14.3 %) and two high-risky polymyxin resistance genes (mcr-4 and mcr-5) were enriched in the industrial effluent receiving area; 2) the HGT potential was higher between resistant microbiome of the industrial effluent and its ERA sediment; and 3) the highest resistome risk was determined in the industrial effluent, and some biocide resistance genes located on high-risky contigs were related to long-term stress of industrial chemicals. These findings highlight the important effects of industrial activities on the development of environmental antimicrobial resistance., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Hypoxia, a characteristic hallmark of solid tumours, restricts the therapeutic effect of photodynamic therapy (PDT) for cancer treatment. To address this issue, a facile and nanosized oxygen (O 2 ) bubble template is established by mixing oxygenated water and water-soluble solvents for guiding hollow polydopamine (HPDA) synthesis, and O 2 is encapsulated in the cavity of HPDA. HPDA with abundant catechol is designed as a carrier for zinc phthalocyanine (ZnPc, a boronic acid modified photosensitizer) via borate ester bonds to fabricate nanomedicine (denoted as HZNPs). The in vitro and in vivo results indicate that O 2 -evolving HZNPs could alleviate tumour hypoxia and enhance PDT-anticancer efficiency. Melanin-like HPDA with a photothermal conversion rate ( η ) of 38.2% shows excellent synergistic photothermal therapy (PTT) efficiency in cancer treatment., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Follicular dendritic cells (FDCs) are a specialized type of stromal cells that exclusively reside in B-cell follicles. When inflammation occurs, the FDC network is reorganized to support germinal center (GC) polarization into the light zone (LZ) and dark zone (DZ). Despite the indispensable role of FDCs in supporting humoral responses, the FDC regulatory requirements remain incompletely defined. In this study, we unexpectedly observed an accumulation of CD169 + subcapsular sinus macrophage (SSM)-derived microvesicles (MVs) in the B-cell zone, which were tightly associated with the FDC network. Interestingly, a selective deposition of CD169 + MVs was detected in both GC LZ FDCs in secondary follicles and on predetermined LZ FDCs in primary follicles. The ablation of CD169 + MVs, resulting from SSM depletion, resulted in significantly decreased expression of LZ-related genes in FDCs. In addition, we found that CD169 + MVs could colocalize with fluorescently tagged antigen-containing immune complexes (ICs), supporting a possible role of CD169 + MVs in transporting antigens to the FDC network. Thus, our data reveal intimate crosstalk between FDCs and SSMs located outside B-cell follicles via SSM-released MVs, providing a novel perspective on the mechanisms underlying the regulation of FDC maturation and polarization., (© 2022 The Authors. European Journal of Immunology published by Wiley-VCH GmbH.)

Microplastics have become global emerging issue and received widespread attention in recent years. Due to their chemical persistence, plastic particles can be broken into smaller items but accumulated for long time in the environment like sediment. However, limited by current detection technologies, the distribution and characteristics of small-sized microplastics in coastal sediment remain uncertain. In this study, we established a new method based on micro-Raman spectroscopy for detecting small-sized microplastics, namely multipoint confocal micro-Raman spectrum scanning (MCmRSS). The MCmRSS was first applied in detecting microplastics in the sediment samples collected from three bays of the East China Sea. The minimum size of microplastics was 4 μm and average microplastics concentration was 91 ± 55 items /g dry weight sediment, with fragment and polyethylene as the most common shape and polymer type, respectively. The spatial variation of microplastics was in accordance with the strength of coastal human activities and marine dynamics. In all the microplastic items, the small-sized ones (<10 μm) accounted for 67%; and the relationship between microplastic concentration and its size followed a power-exponential equation. Compared with previous studies, the number of microplastics in coastal sediments detected by the MCmRSS increased by 2 orders of magnitude, which was benefited from the advantages of multipoint scanning in the fixed identification areas and high resolution of micro-Raman spectrum. Our findings would summon the re-evaluation of the potential risks of small-sized microplastics in the coastal environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Copper-based photosensitizer nanoparticle has high potential clinic translation potency for its extensive physiological effects such as anti-cancer progression, anti-bacteria and accelerate tissue regeneration. However, copper excess or improper coordination can induce toxicity or reduce drug efficacy. To get proper copper-photosensitizer complex nanoparticle, a portion of chlorin e6 covalently conjugated with low molecular weight fish collagen fragments-collage tripeptides (CTPs), and Cu 2+ subsequently triggers CTP-Ce6 conjugates assemble to Cu(II) based CTP-Ce6 nanosphere(CCeC-Ns). CCeC-Ns are 10-20 nm nanoparticles. CCeC-Ns quenched Ce6 fluorescence in aqueous solution and improved longer wavelength light absorbance. It exhibited dramatically higher cellular uptake rates and much more anticancer potency than those of free Ce6 under 660 nm irradiation without obvious dark toxicity in vitro. CCeC-Ns have longer retention time and higher penetrating rate than free Ce6 in tumor spheroid model. CCeC-Ns displayed extremely higher anti-bacterial potency than free Ce6 and sustainable efficacy. It provides a more potent and safer nanodrug for cancer and infection treatment and an idea for highly efficient metal-photosensitizers complexes design., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Transmembrane kinases (TMKs) play important roles in plant growth and signaling cascades of phytohormones. However, its function in the regulation of early leaf senescence (ELS) of plants remains unknown. Here, we report the molecular cloning and functional characterization of the WATER-SOAKED SPOT1 gene which encodes a protein belongs to the TMK family and controls chloroplast development and leaf senescence in rice ( Oryza sativa L.). The water-soaked spot1 ( oswss1 ) mutant displays water-soaked spots which subsequently developed into necrotic symptoms at the tillering stage. Moreover, oswss1 exhibits slightly rolled leaves with irregular epidermal cells, decreased chlorophyll contents, and defective stomata and chloroplasts as compared with the wild type. Map-based cloning revealed that OsWSS1 encodes transmembrane kinase TMK1. Genetic complementary experiments verified that a Leu396Pro amino acid substitution, residing in the highly conserved region of leucine-rich repeat (LRR) domain, was responsible for the phenotypes of oswss1 . OsWSS1 was constitutively expressed in all tissues and its encoded protein is localized to the plasma membrane. Mutation of OsWSS1 led to hyper-accumulation of reactive oxygen species (ROS), more severe DNA fragmentation, and cell death than that of the wild-type control. In addition, we found that the expression of senescence-associated genes (SAGs) was significantly higher, while the expression of genes associated with chloroplast development and photosynthesis was significantly downregulated in oswss1 as compared with the wild type. Taken together, our results demonstrated that OsWSS1, a member of TMKs, plays a vital role in the regulation of ROS homeostasis, chloroplast development, and leaf senescence in rice., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Xu, Ji, Wang, Xu, Wang, Zheng, Tang, Wei, Zhao and Zhao.)

Mangrove ecosystems play an important role in global carbon budget, however, the quantitative relationships between environmental drivers and productivity in these forests remain poorly understood. This study presented a remote sensing (RS)-based productivity model to estimate the light use efficiency (LUE) and gross primary production (GPP) of mangrove forests in China. Firstly, LUE model considered the effects of tidal inundation and therefore involved sea surface temperature (SST) and salinity as environmental scalars. Secondly, the downscaling effect of photosynthetic active radiation (PAR) on the mangrove LUE was quantified according to different PAR values. Thirdly, the maximum LUE varied with temperature and was therefore determined based on the response of daytime net ecosystem exchange and PAR at different temperatures. Lastly, GPP was estimated by combining the LUE model with the fraction of absorbed photosynthetically active radiation from Sentinel-2 images. The results showed that the LUE model developed for mangrove forests has higher overall accuracy (RMSE = 0.0051, R 2  = 0.64) than the terrestrial model (RMSE = 0.0220, R 2  = 0.24). The main environmental stressor for the photosynthesis of mangrove forests in China was PAR. The estimated GPP was, in general, in agreement with the in-situ measurement from the two carbon flux towers. Compared to the MODIS GPP product, the derived GPP had higher accuracy, with RMSE improving from 39.09 to 19.05 g C/m 2 /8 days in 2012, and from 33.76 to 19.51 g C/m 2 /8 days in 2015. The spatiotemporal distributions of the mangrove GPP revealed that GPP was most strongly controlled by environmental conditions, especially temperature and PAR, as well as the distribution of mangroves. These results demonstrate the potential of the RS-based productivity model for scaling up GPP in mangrove forests, a key to explore the carbon cycle of mangrove ecosystems at national and global scales., (© 2022. The Author(s).)

2,4,6-Trichlorophenol (2,4,6-TCP) is extensively consumed in industrial production and may cause environmental damages. The effect of halide ions on the decomposition of 2,4,6-TCP has often been overlooked. In this study, the bromide ion was found to have a stronger negative impact on 2,4,6-TCP degradation than chloride ion in the O 3 system, and led to the formation of adsorbable organic halogens (AOX). Kinetic modeling demonstrated that the concentration of various radicals was largely depended on the solution pH, and stronger basicity not only contributed to the mineralization of 2,4,6-TCP, but also inhibited the formation of halogenated by-products. Combining the intermediate identification and quantum chemical calculation, the degradation pathways of 2,4,6-TCP during ozone oxidation process were proposed. The toxicity test and ECOSAR simulation demonstrated that the acute toxicity of some 2,4,6-TCP degradation intermediates was relatively higher than their parent compound. With high concentrations of halide ions, the ozone-treated solution showed greater toxicity than the originator 2,4,6-TCP solution. These results illustrate that the ozone treatment of the halide-containing wastewater may cause potential ecological hazards and its application needs to be more cautious., (Copyright © 2021. Published by Elsevier Ltd.)

Climate change has become a great challenge for humanity. However, the current climate change mitigation measures, primarily concentrate on land, more or less neglecting the vital role of the ocean-based solutions. While the ocean is a crucial regulator of the global climate, ocean-based solutions could also play an essential role in climate change mitigation and policymaking. This paper developed an Ocean-based Solutions Carbon Reduction Assessment Model (OSCRAM) that addresses coastal ecosystems, ocean energy, marine transportation, fishery, and seabed to estimate the oceanic contribution to climate change mitigation. It has been applied to evaluate the capacity of carbon emission reduction through oceans in China. We found that the total contribution for carbon emission reduction was about 6.86 Tg CO 2 per year, and it may reach 139.39 Tg in 2030 under the target scenario. The results indicated that the ocean has huge potential to reduce carbon emissions. The development of marine energy and low-carbon marine shipping may have more potential for emission reduction in China, and the government should also protect and restore coastal wetlands for their enormous carbon storage. It can also provide a reference for the globe and other nations in achieving emission reduction goals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

We investigate the topology optimization of geometric phase metasurfaces for wide-angle and high-efficiency deflection, where adjoint-based multi-object optimization approach is adopted to improve the absolute efficiency while maintaining the polarization conversion characteristic of geometric phase metasurfaces. We show that, for the initially discrete geometric phase metasurfaces with different materials and working wavelengths, the topology shapes gradually evolve from discrete structures to quasi-continuous arrangements with the increment of optimization iteration operations. More importantly, the finally optimized metasurfaces manifest as catenary-like structure, providing significant improvements of absolute efficiency. Furthermore, for the initial structure with catenary distribution, the corresponding optimized metasurface also has a catenary-like topology shape. Our results on the topology-optimized geometric phase metasurfaces reveal that, from the perspective of numerical optimization, the continuous catenary metasurfaces is superior to the discrete geometric phase metasurfaces.

Lesion mimic mutants (LMMs) are ideal materials for studying programmed cell death and defense response in plants. Here we report investigations on two LMMs ( msl-1 and msl-2 ) from the indica rice cultivar JG30 treated by ethyl methyl sulfone. Both of the mutants showed similar mosaic spot lesions at seedling stage, but they displayed different phenotypes along with development of the plants. At tillering stage, larger orange spots appeared on leaves of msl-2 , while only small reddish-brown spots exhibit on leaves of msl-1 . At heading stage, the msl-2 plants were completely dead, while the msl-1 plants were still alive even if showed apparent premature senility. For both the mutants, the mosaic spot lesion formation was induced by light; DAB and trypan blue staining showed a large amount of hydrogen peroxide accumulated at the lesion sites, accompanied by a large number of cell death. Consequently, reactive oxygen species were enriched in leaves of the mutants; SOD and CAT activities in the scavenging enzyme system were decreased compared with the wild type. In addition, degraded chloroplasts, decreased photosynthetic pigment content, down-regulated expression of genes associated with chloroplast synthesis/photosynthesis and up-regulated expression of genes related to senescence were detected in the mutants, but the abnormality of msl-2 was more serious than that of msl-1 in general. Genetic analysis and map-based cloning revealed that the lesion mimic and premature senescence traits of both the mutants were controlled by recessive mutated alleles of the SL (Sekiguchi lesion) gene, which encodes the CYP71P1 protein belonging to cytochrome P450 monooxygenase family. The difference of mutation sites and mutation types (SNP-caused single amino acid change and SNP-caused early termination of translation) led to the different phenotypes in severity between msl-1 and msl-2 . Taken together, this work revealed that the CYP71P1 is involved in regulation of both premature senescence and cell death in rice, and its different mutation sites and mutation types could cause different phenotypes in terms of severity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zheng, Xu, Wang, Tang, Wei, Ji, Wang and Zhao.)

Antibiotics have raised significant concern as emerging pollutants for their increasing consumption, persistent input, and potential threat to ecological environment. Due to low concentrations and various types in coastal water, simultaneous quantification of all kinds of antibiotics is time-consuming and costly. In order to make antibiotic regular monitoring in coastal water possible, identifying the priority antibiotics in the environment is essential. Here, a method for screening the priority antibiotics in coastal water was proposed, considering individual antibiotic concentration, the positive correlation between individual and total antibiotic concentration, the detection frequency, and obvious ecological risk. Taking coastal water of the East China Sea as an example, on a list of 77 target antibiotics, 7 (SMX, TMP, SCP, SMP, CNX, ATM, and ETM) and 4 (SMX, SCP, SMP, and CNX) antibiotics were selected to be the priority antibiotics in 2017 and 2018, respectively. Furthermore, the 4 priority antibiotics in 2018 were all involved in the 7 priority antibiotics in 2017. The sum of the priority antibiotic concentrations accounted for 0.8% and 23.2% of total antibiotic concentrations, and the sum of their RQ accounted for 69.2% and 66.8% of total RQ values in 2017 and 2018, respectively. Among the above 7 priority antibiotics, ATM is mainly used in human clinical, SMX, SCP, and SMP are mainly consumed in veterinary medicine, TMP, CNX, and ETM are commonly used for humans and animals. The proposed method might provide an important reference for the monitoring and management of antibiotic pollution in coastal water., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The distribution of 77 antibiotics in the coastal water and sediment from 3 bays of the East China Sea was investigated. There were 43 and 25 antibiotics detected with total concentrations of 30.8-2106.1 ng/L and 2.2-99.9 ng/g in water and sediment, respectively. Approximately 83.0% and 85.4% of the individual antibiotic concentrations were lower than 5.0 ng/L in water and 1.0 ng/g in sediment. Clindamycin (1.2-1507.9 ng/L, mean 183.8 ng/L) and erythromycin (ND-45.2 ng/g, mean 3.4 ng/g) were the most abundant in water and sediment, respectively. Ecological risk assessment revealed that the joint toxicity was enhanced when multiple antibiotics were present simultaneously. A decrease in the total antibiotic concentration and the ecological risk in water was observed from nearshore to offshore. Three antibiotics (sulfamethoxypyridazine, sulfamethoxazole and cinoxacin) were selected to be prioritized based on ecological risks for antibiotics monitoring and management of the coastal water in the East China Sea., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

This study presented a novel alternating-anode electrochemical system (AAES) based on single electrolytic cell for the treatment of nitrogen-containing refractory organic wastewater (NOW). The core of AAES lies in the alternating working of iron anode and DSA anode to integrate different electrochemical processes. The biologically treated landfill leachate (BTLL) was selected as a practical NOW for assessing the performance of AAES. The results indicated that after 140 min of electrolytic reaction, the removal efficiency of chemical oxygen demand and total nitrogen (TN) using AAES was found to be 76.9 and 98.9%, respectively. The main component of dissolved organic matter (DOM) in BTLL included humic-like substances, which could be degraded into small-molecule DOM, such as fulvic-like substances and protein-like substances, by available chlorine and hydroxyl radicals present in AAES. Cathode reduction (NO x - -N → NH 4 + -N and N 2 ) under iron anode and indirect oxidation (NH 4 + -N → N 2 ) under DSA anode were the main pathways to remove TN from NOW. Owing to the redox conditions created by the alternating anodes, the main stable crystalline forms of precipitates obtained from AAES were Fe 3 O 4 and γ-Fe 2 O 3 , which could be separated by using the external magnetic field. The findings of this study may provide a feasible solution for the advanced electrochemical treatment of NOW in a single electrolytic cell as well as rapid separation of precipitates., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objective: To investigate the effect of Golgi phosphoprotein 3 (Golph3) on paclitaxel- induced apoptosis and autophagy in HeLa cells., Methods: HeLa cells were transfected with a lentiviral vector expressing Golph3 or a small interfering RNA (siRNA) targeting Golph3 for up-regulation or down-regulation of Golph3 which was verified by Western blotting. The autophagic bodies in the cells were observed using transmission electron microscopy. The expression of autophagy markers p62 and LC3 were detected using Western blotting, and the cell apoptosis was examined by PI/Anexin V-FITC double staining and flow cytometry. The effects of blocking autophagy was evaluated by treatment of the cells with the autophagy inhibitor 3-MA., Results: Transmission electron microscopy showed that the lentivirus-mediated overexpression of Golph3 significantly increased the number of autophagic bodies and interference of Golph3 expression significantly decreased autophagic bodies in HeLa cells. Western blotting showed that Golph3 overexpression caused an increased expression of LC3 and decreased the accumulation of p62 in the cells, and interference of Golph3 resulted in the reverse changes. The cell apoptosis induced by paclitaxel was significantly decreased in Golph3-overexpressing HeLa cells and increased in the cells with Golph3 knockdown ( P >0.01). Treatment with 3-MA alone did not obviously affect HeLa cell apoptosis, but in cells with Golph3 knockdown, 3-MA significantly enhanced paclitaxel-induced apoptosis ( P >0.01)., Conclusions: Up-regulation of Golph3 promotes autophagy and inhibits paclitaxel-induced apoptosis, whereas suppression of Golph3 inhibits autophagy and enhances paclitaxel- induced apoptosis in HeLa cells.

Minimal hepatic encephalopathy (MHE) is characterized as cognitive deficits including memory and learning dysfunctions after liver injuries or hepatic diseases. Our understandings of neurological mechanisms of MHE-associated cognitive syndromes, however, are far from complete. In the current study we generated a mouse MHE model by repetitive administrations of thioacetamide (TAA), which induced hyperammonemia plus elevated proinflammatory cytokines in both the general circulation and motor cortex. MHE mice presented prominent motor learning deficits, which were associated with excess dendritic spine pruning in the motor cortex under 2-photon in vivo microscopy. The pharmaceutical blockade of glucocorticoid receptor or suppression of its biosynthesis further rescued motor learning deficits and synaptic protein loss. Moreover, MHE mice presented microglial activation, which can be alleviated after glucocorticoid pathway inhibition. In sum, our data demonstrates corticosterone-induced microglial activation, synaptic over-pruning and motor learning impairments in MHE, providing new insights for MHE pathogenesis and potential targets of clinical interventions., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The humic-like substances were the main organic components in most wastewater (e.g. domestic sewage, toilet wastewater and landfill leachate). Two types of actual humic-like substances (fulvic acid (FA) and biologically treated landfill leachate (BTLL)) were selected to describe the changes in the properties of humic-like substances (complexation ability, aromaticity and mobility) during electrochemical oxidation. Meanwhile, the acute cytotoxicity of FA and BTLL was also tested by acute toxicological test of luminescent bacteria. The results showed that the consumption of coordinating groups such as phenolic groups and hydrogen bonds reduced the complexation ability of FA and BTLL. The functional groups were degraded with the removal order of quinone group, phenolic group and aromatic group, and finally realized the molecular saturation and aromaticity decrease for humic-like substances. The mobility of FA and BTLL was decreased because of the enhancement of hydrophobicity during electrolysis process. Furthermore, the available chlorine produced during electrochemical oxidation was the main acute cytotoxicity substance, therefore, it is necessary to remove it before discharge in order to reduce ecological risks. This study provides a basis for understanding and evaluating the electrochemical degradation process of humic-like substances in detail., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Physical exercise improves learning and memory, but little in vivo evidence has been provided to illustrate the molecular mechanisms. Here, we show that chronic treadmill exercise activates the mechanistic target of rapamycin (mTOR) pathway in mouse motor cortex. Both ex vivo and in vivo recordings suggest that mTOR activation leads to potentiated postsynaptic excitation and enhanced neuronal activity of layer 5 pyramidal neurons after exercise, in association with increased oligodendrogenesis and axonal myelination. Exercise training also increases dendritic spine formation and motor learning. Together, exercise activates mTOR pathway, which is necessary for spinogenesis, neuronal activation, and axonal myelination leading to improved motor learning. This model provides new insights for neural network adaptations through exercises and supports the intervention of cognitive deficits using exercise training.

Autistic spectral disorder (ASD) is a prevalent neurodevelopmental disease that affects multiple brain regions. Both clinical and animal studies have revealed the possible involvement of the cerebellum in ASD pathology. In this study, we generated a rodent ASD model through a single prenatal administration of valproic acid (VPA) into pregnant mice, followed by cerebellar morphological and functional studies of the offspring. Behavioral studies showed that VPA exposure led to retardation of critical motor reflexes in juveniles and impaired learning in a tone-conditioned complex motor task in adults. These behavioral phenotypes were associated with premature migration and excess apoptosis of the granular cell (GC) precursor in the cerebellar cortex during the early postnatal period, and the decreased cell density and impaired dendritic arborization of the Purkinje neurons. On acute cerebellar slices, suppressed synaptic transmission of the Purkinje cells were reported in the VPA-treated mice. In summary, converging evidence from anatomical, electrophysiological and behavioral abnormalities in the VPA-treated mice suggest cerebellar pathology in ASD and indicate the potential values of motor dysfunction in the early diagnosis of ASD.

Metabolic programs are crucial for regulatory T (T reg) cell stability and function, but the underlying mechanisms that regulate T reg cell metabolism are elusive. Here, we report that lysosomal TRAF3IP3 acts as a pivotal regulator in the maintenance of T reg cell metabolic fitness. T reg-specific deletion of Traf3ip3 impairs T reg cell function, causing the development of inflammatory disorders and stronger antitumor T cell responses in mice. Excessive mechanistic target of rapamycin complex 1 (mTORC1)-mediated hyper-glycolytic metabolism is responsible for the instability of TRAF3IP3-deficient T reg cells. Mechanistically, TRAF3IP3 restricts mTORC1 signaling by recruiting the serine-threonine phosphatase catalytic subunit (PP2Ac) to the lysosome, thereby facilitating the interaction of PP2Ac with the mTORC1 component Raptor. Our results define TRAF3IP3 as a metabolic regulator in T reg cell stability and function and suggest a lysosome-specific mTORC1 signaling mechanism that regulates T reg cell metabolism., (© 2018 Yu et al.)

Blastocystis is a common protozoan found in the surveys of human and animal fecal specimens. Extensive genetic diversity has been observed within the genus Blastocystis. At least 17 subtypes (ST) have been identified in mammals and birds, nine of which (ST1 to ST9) have been identified in humans. In China, although there have been a few reports on Blastocystis infection in humans and many animal species, no epidemiological data are available in either humans or animals in northeastern China's Heilongjiang Province. To determine infection rates and subtype distributions of Blastocystis in pigs, cattle, sheep and goats, to understand genetic characterizations and to assess zoonotic possibility of Blastocystis isolates, 337 fecal specimens from livestock (68 from pigs, 147 from cattle, 109 from sheep and 13 from goats) were collected in northeastern China's Heilongjiang Province. Each of them was detected for Blastocystis by PCR amplification of the partial SSU rRNA gene. An average infection rate of Blastocystis was 7.7% (26/337), and the highest infection rate was found in 9.5% (14/147) in cattle, followed by 8.8% (6/68) in pigs and 5.5% (6/109) in sheep. There was an absence of Blastocystis in goats. Five Blastocystis subtypes were identified: ST5 (n=6) in pigs; ST3 (n=2), ST10 (n=10) and ST14 (n=2) in cattle; ST1 (n=1), ST5 (n=1), ST10 (n=3) and ST14 (n=1) in sheep. 38.5% (10/26) of Blastocystis isolates belong to potentially zoonotic subtypes based on the previous findings of ST1, ST3 and ST5 in humans. This is the first report of Blastocystis in pigs, cattle and sheep in northeastern China's Heilongjiang Province. ST1, ST5 and ST14 were identified in sheep for the first time. Due to the low infection rate of Blastocystis and the small percentage of potentially zoonotic subtypes in these livestock, there is a minimal risk of zoonotic transmission of Blastocystis., (Copyright © 2017 Elsevier B.V. All rights reserved.)

China is facing intense coastal eutrophication. Large-scale seaweed aquaculture in China is popular, now accounting for over 2/3's of global production. Here, we estimate the nutrient removal capability of large-scale Chinese seaweed farms to determine its significance in mitigating eutrophication. We combined estimates of yield and nutrient concentration of Chinese seaweed aquaculture to quantify that one hectare of seaweed aquaculture removes the equivalent nutrient inputs entering 17.8 ha for nitrogen and 126.7 ha for phosphorus of Chinese coastal waters, respectively. Chinese seaweed aquaculture annually removes approximately 75,000 t nitrogen and 9,500 t phosphorus. Whereas removal of the total N inputs to Chinese coastal waters requires a seaweed farming area 17 times larger than the extant area, one and a half times more of the seaweed area would be able to remove close to 100% of the P inputs. With the current growth rate of seaweed aquaculture, we project this industry will remove 100% of the current phosphorus inputs to Chinese coastal waters by 2026. Hence, seaweed aquaculture already plays a hitherto unrealized role in mitigating coastal eutrophication, a role that may be greatly expanded with future growth of seaweed aquaculture.

Viral infection causes host cells to produce type I IFNs, which play a critical role in viral clearance. IFN regulatory factor (IRF) 7 is the master regulator of type I IFN-dependent immune responses. In this article, we report that N-Myc and STATs interactor (Nmi), a Sendai virus-inducible protein, interacted with IRF7 and inhibited virus-triggered type I IFN production. The overexpression of Nmi inhibited the Sendai virus-triggered induction of type I IFNs, whereas the knockdown of Nmi promoted IFN production. Furthermore, the enhanced production of IFNs resulting from Nmi knockdown was sufficient to protect cells from infection by vesicular stomatitis virus. In addition, Nmi was found to promote the K48-linked ubiquitination of IRF7 and the proteasome-dependent degradation of this protein. Finally, an impairment of antiviral responses is also detectable in Nmi-transgenic mice. These findings suggest that Nmi is a negative regulator of the virus-triggered induction of type I IFNs that targets IRF7.

Type 2 helper T cells (T(H)2) are critically involved in allergies and asthma. Here we demonstrate that extracellular matrix protein-1 (ECM1) is highly and selectively expressed in T(H)2 cells. ECM1 deficiency caused impaired T(H)2 responses and reduced allergic airway inflammation in vivo. Functional analysis demonstrated that although the T(H)2 polarization of ECM1-deficient cells was unimpaired, these cells had a defect in migration and were retained in peripheral lymphoid organs. This was associated with reduced expression of KLF2 and S1P(1). We also found that ECM1 could directly bind the interleukin-2 (IL-2) receptor to inhibit IL-2 signaling and activate S1P(1) expression. Our data identify a previously unknown function of ECM1 in regulating T(H)2 cell migration through control of KLF2 and S1P(1) expression.

Perforin is a pore-forming protein engaged mainly in mediating target T cell death and is employed by cytotoxic T lymphocytes (CTLs) and natural killer cells. However, whether it also plays a role in conventional CD4(+) T cell function remains unclear. Here we report that in perforin-deficient (PKO) mice, CD4(+) T cells are hyperproliferative in response to T cell receptor (TCR) stimulation. This feature of hyperproliferation is accompanied by the enhancement both in cell division and in IL-2 secretion. It seems that the perforin deficiency does not influence T cell development in thymus spleen and lymph node. In vivo, perforin deficiency results in increased antigen-specific T cell proliferation and antibody production. Furthermore, PKO mice are more susceptible to experimental autoimmune uveitis. To address the molecular mechanism, we found that after TCR stimulation, CD4(+) T cells from PKO mice display an increased intracellular calcium flux and subsequently enhance activation of transcription factor NFAT1. Our results indicate that perforin plays a negative role in regulating CD4(+) T cell activation and immune response by affecting TCR-dependent Ca(2+) signaling.