Discovery of small molecules for autophagy-lysosome degradation of immune checkpoint proteins.

Targeted protein degradation (TPD) technologies, particularly proteolysis targeting chimeras (PROTACs), have emerged as a promising branch of targeted therapy. Current ubiquitin-proteasome-dependent TPD technologies are limited to targeting intracellular proteins. Although the blockade of immune checkpoints has achieved great clinical success, most immune checkpoints are transmembrane proteins, which are difficult to be ubiquitinated and degraded by PROTACs. Herein, we developed a novel discovery strategy of bifunctional small molecules, which could mediate autophagy-lysosome degradation of immune checkpoints. F-1 was demonstrated to activate the autophagy-lysosome system, and conjugation of F-1 with inhibitors targeting programmed cell death-ligand 1 (PD-L1) or V-domain Ig suppressor of T-cell activation (VISTA) generated a new class of small molecules that effectively induce the degradation of PD-L1 or VISTA in tumor cells. The most promising PD-L1 degrader B3 significantly induced PD-L1 degradation in RKO cells through the autophagy-lysosome system and exhibited good tumor-inhibiting effects in vivo. Our work could expand the development of degraders targeting immune checkpoints and provide a promising discovery strategy for future autophagy-lysosome targeting degradation technology.

Review of Acoustic Agglomeration Technology Research.

Acoustic agglomeration is employed as a precursor technique that modifies the sound field of fine particles to increase their size, thereby facilitating more efficient emission control. This paper reviews progress in the field of acoustic agglomeration technology, clarifies the mechanisms at play within the acoustic agglomeration process, and outlines its applicability in both gas-liquid and gas-solid phases. Furthermore, it analyzes the factors impacting the efficacy of acoustic agglomeration, summarizes the numerical simulation research of acoustic agglomeration, and proposes directions for technological enhancement.

Real-Time Temperature Monitoring of Lithium Batteries Based on Ultrasonic Technology.

Electrochemical energy storage stations serve as an important means of load regulation, and their proportion has been increasing year by year. The temperature monitoring of lithium batteries necessitates heightened criteria. Ultrasonic thermometry, based on its noncontact measurement characteristics, is an ideal method for monitoring the internal temperature of lithium batteries. In this study, temperature and ultrasonic time delay measurement experiments were conducted on 18650 lithium batteries and laminated and wound lithium batteries to obtain the corresponding relationship between temperature and time delay and validate the temperature measurement for the same type of battery. The experimental results show that (1) the ultrasonic temperature measurement technique exhibits a relatively large error when used for 18650 Li-ion batteries under experimental conditions; (2) in the experiments on laminated and wound soft-pack lithium batteries, the relationship between temperature and time delay exhibits a nonlinear characteristic; and (3) under the experimental conditions, the ultrasonic temperature measurement errors were ±1.1 °C for stacked Li-ion batteries and ±1.4 °C for wound Li-ion batteries.

Advances in colorimetric aptasensors for heavy metal ion detection utilizing nanomaterials: a comprehensive review.

Heavy metal ion contamination poses significant environmental and health risks, necessitating rapid and efficient detection methods. In the last decade, colorimetric aptasensors have emerged as powerful tools for heavy metal ion detection, owing to their notable attributes such as high specificity, facile synthesis, adaptability to modifications, long-term stability, and heightened sensitivity. This comprehensive overview summarizes the key developments in this field over the past ten years. It discusses the principles, design strategies, and innovative techniques employed in colorimetric aptasensors using nanomaterials. Recent advancements in enhancing sensitivity, selectivity, and on-site applicability are highlighted. The review also presents application studies of successful heavy metal ion detection using colorimetric aptasensors, underlining their potential for environmental monitoring and health protection. Finally, future directions and challenges in the continued evolution of these aptasensors are outlined.

Genome-wide identification and analysis of the MAPKK gene family in Chinese mitten crab (Eriocheir sinensis) and its response to bacterial challenge.

Protein kinases of the MAPK cascade family (MAPKKK-MAPKK-MAPK) play an important role in the growth and development of organisms and their response to environmental stress. The MAPKK gene families in the Chinese mitten crab Eriocheir sinensis have never been systematically analyzed. We identified four MAPKK family genes, EsMEK, EsMAPKK4, EsMAPKK6, and EsMAPKK7, in E. sinensis and analyzed their molecular features and expression patterns. All four MAPKK genes are composed of multiple exons and introns, all have a conserved domain, and all have 10 conserved motifs (except EsMEK and EsMAPKK7 which are missing motif 10). The four MAPKK genes are on four different chromosomes and have no gene duplications, and the results of phylogenetic tree analysis indicate that the ESMAPKK gene family is highly conserved evolutionarily. The EsMAPKK genes were widely expressed in all the examined tissues with higher expression in hemocytes, hepatopancreas, and gills. Notably, EsMAPKK6 was also highly expressed in the ovary. Vibrio parahaemolyticus infection significantly increased the mRNA levels of the EsMAPKK genes in hemocytes. Further disruption of the EsMAPKK gene family expression affects the expression levels of multiple antimicrobial peptides in hemocytes. Our experimental results provide a starting point for a more in-depth study of the innate immunity functional roles of members of the MAPKK gene families in E. sinensis.

Effects of the tumor necrosis factor on hemocyte proliferation and bacterial infection in Chinese mitten crab (Eriocheir sinensis).

Tumor necrosis factor (TNF) is an important cytokine that can regulate a variety of cellular responses by binding tumor necrosis factor receptor (TNFR). We studied whether the TNF of Eriocheir sinensis can regulate hemocyte proliferation. The results showed that the EsTNF and EsTNFR were constitutively expressed in all tested tissues, including the heart, hepatopancreas, muscles, gills, stomachs, intestines, and hemocytes. We found that low levels of EsTNF and EsTNFR transcripts were present in hemocytes. The gene expression levels were significantly increased in the hemocytes after being stimulated by Staphylococcus aureus or Vibrio parahaemolyticus. We also found some genes related to cell proliferation were expressed at a higher level in pulsing rTNF-stimulated hemocytes compared with the control group. We also knocked down the EsTNFR gene with RNAi technology. The results showed that the expression level of these genes related to cell proliferation was significantly down-regulated compared with the control group when the TNF does not bind TNFR. We used Edu technology to repeat the above experiments and the results were similar. Compared with the control group, the hemocytes stimulated by rTNF showed more significant proliferation, and the proliferation rate was significantly down-regulated after knocking down the EsTNFR gene. Therefore, we indicate that TNF binding TNFR can affect the proliferation of E. sinensis hemocytes, which might be manifested by affecting the expression of some proliferation-related genes.

Improving aptamer performance: key factors and strategies.

Aptamers are functional single-stranded oligonucleotide fragments isolated from randomized libraries by Systematic Evolution of Ligands by Exponential Enrichment (SELEX), exhibiting excellent affinity and specificity toward targets. Compared with traditional antibody reagents, aptamers display many desirable properties, such as low variation and high flexibility, and they are suitable for artificial and large-scale synthesis. These advantages make aptamers have a broad application potential ranging from biosensors, bioimaging to therapeutics and other areas of application. However, the overall performance of aptamer pre-selected by SELEX screening is far from being satisfactory. To improve aptamer performance and applicability, various post-SELEX optimization methods have been developed in the last decade. In this review, we first discuss the key factors that influence the performance or properties of aptamers, and then we summarize the key strategies of post-SELEX optimization which have been successfully used to improve aptamer performance, such as truncation, extension, mutagenesis and modification, splitting, and multivalent integration. This review shall provide a comprehensive summary and discussion of post-SELEX optimization methods developed in recent years. Moreover, by discussing the mechanism of each approach, we highlight the importance of choosing the proper method to perform post-SELEX optimization.

Immune functions of the Dscam extracellular variable region in Chinese mitten crab.

In arthropods, there is only a single copy of Down Syndrome Cell Adhesion Molecule (Dscam) in the genome, but it can exist as numerous splice variants. There are three hypervariable exons in the extracellular domain and one hypervariable exon in the transmembrane domain. In Chinese mitten crab (Eriocheir sinensis), exons 4, 6 and 14 can produce 25, 34 and 18 alternative splice variants, respectively. In this study, through Illumina sequencing, we identified additional splice variants for exons 6 and 14, hence there may be > 50,000 Dscam protein variants. Sequencing of exons 4, 6 and 14 showed that alternative splicing was altered after bacterial stimulation. Therefore, we expressed and purified the extracellular variable region of Dscam (EsDscam-Ig1-Ig7). Exons 4.3, 6.46 and 14.18, three variable exons of the recombinant protein, were randomly selected. The functions of EsDscam-Ig1-Ig7 in immune defences of E. sinensis were subsequently explored. EsDscam-Ig1-Ig7 was discovered to bind to both Gram-positive Staphylococcus aureus and Gram-negative Vibrio parahaemolyticus, but it did not exhibit antibacterial activity. By promoting hemocyte phagocytosis and bacterial removal, EsDscam-Ig1-Ig7 can also shield the host from bacterial infection. The findings highlight the immunological activities of Dscam alternative splicing and reveal the potential for many more Dscam isoforms than were previously predicted in E. sinensis.

A novel peroxymonosulfate activation process by single-atom iron catalyst from waste biomass for efficient singlet oxygen-mediated degradation of organic pollutants.

Single-atom dispersed catalysts (SACs) have gained considerable attention in organic contaminants remediation due to their superior reactivity and stability. However, the complex and costly synthesis processes limit their practical applications in environmental protection. Herein, a facile and cost-effective single-atom iron catalyst (Fe-SA/NC) anchored on nitrogen-doped porous carbon was first fabricated by using waste biomass as a carbon source. The Fe-SA/NC catalyst exhibited outstanding performance with a high turnover frequency of 1.72 min-1 toward antibiotics degradation via peroxymonosulfate activation. ECOSAR program and algae growth experiments demonstrated that the byproducts produced during the sulfamethoxazole degradation process were not detrimental to the aquatic environment. Radical quenching and electron paramagnetic resonance experiments revealed that Fe-SA/NC remarkably promoted 1O2 production in PMS-assisted reaction, and thus 1O2 contributed as much as 78.77% to sulfamethoxazole degradation. As indicated by experiment and density functional theory (DFT) calculations, FeN2O2 configuration serves as the active site. DFT calculations further presented the most rational generation route of 1O2 as PMS→OH* â†’O* â†’1O2. We also designed Fe-SA/NC embedded spherical pellets for contaminants elimination at the device level. This study offers new insights into the synthesis of SACs from waste biomass and their practical application in environmental remediation.

Deubiquitinase cylindromatosis (CYLD) regulates antibacterial immunity and apoptosis in Chinese mitten crab (Eriocheir sinensis).

Ubiquitination and deubiquitination of target proteins is an important mechanism for cells to rapidly respond to changes in the external environment. The deubiquitinase, cylindromatosis (CYLD), is a tumor suppressor protein. CYLD from Drosophila melanogaster participates in the antimicrobial immune response. In vertebrates, CYLD also regulates bacterial-induced apoptosis. However, whether CYLD can regulate the bacterial-induced innate immune response in crustaceans is unknown. In the present study, we reported the identification and cloning of CYLD in Chinese mitten crab, Eriocheir sinensis. Quantitative real-time reverse transcription polymerase chain reaction analysis showed that EsCYLD was widely expressed in all the examined tissues and was upregulated in the hemolymph after Vibrio parahaemolyticus challenge. Knockdown of EsCYLD in hemocytes promoted the cytoplasm-to-nucleus translocation of transcription factor Relish under V. parahaemolyticus stimulation and increased the expression of corresponding antimicrobial peptides. In vivo, silencing of EsCYLD promoted the removal of bacteria from the crabs and enhanced their survival. In addition, interfering with EsCYLD expression inhibited apoptosis of crab hemocytes caused by V. parahaemolyticus stimulation. In summary, our findings revealed that EsCYLD negatively regulates the nuclear translocation of Relish to affect the expression of corresponding antimicrobial peptides and regulates the apoptosis of crab hemocytes, thus indirectly participating in the innate immunity of E. sinensis.

Integrinß1/FAK/ERK signalling pathway is essential for Chinese mitten crab Eriocheir sinensis hemocyte survival.

Integrins are cellular adhesion molecules that mediate cell-cell, cell-extracellular matrix, and cell-pathogen interactions. Integrins can stimulate various signaling pathways by binding to different ligands, thereby exerting immunological functions. While integrins have been found to primarily play a role in bacterial agglutination, phagocytosis, and inhibition of apoptosis in invertebrates, the specific signaling pathway and mechanism of action remain unclear. In vertebrates, ß1 integrin and extracellular matrix interactions can associate with focal adhesion kinase (FAK) to initiate MAPK/ERK signaling and regulate cell survival; however, in invertebrates (e.g., Chinese mitten crab), the mechanisms of integrins are poorly understood. The purpose of this study was to investigate whether integrinß1/FAK activation of the MAPK/ERK pathway regulates hemocyte survival and the associated mechanism. Treatment with an integrinß1 inhibitor RGD (a conserved tripeptide Arg-Gly-Asp), decreased the levels of FAK and ERK expression and phosphorylation, followed by an intensification of apoptosis. Similar results were obtained following siRNA knockdown of integrinß1 expression. We further found that the attenuation of ERK phosphorylation enhanced the level of Caspase-3 expression. Together, these findings suggest that integrinß1 activates the FAK/ERK signaling cascade and is involved in the survival of Chinese mitten crab hemocytes.

The extracellular non-variable region of Dscam promotes bacterial clearance by promoting phagocytosis of hemocytes in Eriocheir sinensis.

As the most typical example of mRNA variable splicing, Down Syndrome Cell Adhesion Molecule (Dscam) can produce a large number of mRNA isomers. It plays an important role not only in the nervous system, but also in the immune system. In Eriocheir sinensis, the extracellular region of Dscam has three variable domains, which can produce 25, 34 and 18 exons and encode the N-terminal region of immunoglobulin (Ig) 2 and Ig3 domains, and the entire Ig7 domain, respectively. In addition to three variable domains, the extracellular non-variable region of Dscam also includes many Ig domains and fibronectin type III (FNIII) domains. However, the role of the extracellular non-variable region function of Dscam in the immune defense of E. sinensis is unclear. In this study, we focused on the role of the extracellular non-variable region of Dscam in crab immune defense. The results indicate that the extracellular non-variable region of Dscam can bind bacteria and has bacteriostatic function. At the same time, the extracellular non-variable region of Dscam can also directly promote bacterial clearance by promoting phagocytosis of hemocytes.

One-step synthesis of highly fluorescent carbon dots as fluorescence sensors for the parallel detection of cadmium and mercury ions.

Cadmium (Cd2+) and mercury ions (Hg2+) are essential for the quality control of food samples because of their serious toxicity to human health, but the effective and simple strategy for their parallel detection remains challenging. In this paper, a rapid and simple parallel detection method for Cd2+ and Hg2+ was developed using carbon dots (CDs) as fluorescent sensors. A one-step hydrothermal method with a single precursor l-arginine as both the carbon and nitrogen sources was employed to prepare nitrogen-doped CDs (N-CDs). N-CDs exhibited a uniform particle size and excitation-independent fluorescence emission. The maximum emission wavelength of N-CDs was observed at 354 nm with the excitation wavelength at 295 nm. The quantum yield of N-CDs reached as high as 71.6% in water. By using sodium diphosphate and phytic acid as masking agents, the fluorescent sensor can be quenched by Cd2+ and Hg2+ in the linear range of 0-26.8 µM and 0-49.9 µM within 5 min. Other common ions in farm products showed no significant effect on the fluorescence intensity of the sensing system. The results demonstrated that the sensing system had good selectivity and sensitivity for Cd2+ and Hg2+. The detection limits for Cd2+ and Hg2+ were 0.20 and 0.188 µM, respectively. In addition, the fluorescent sensor had been successfully applied for the detection of Cd2+ and Hg2+ in fruits and vegetables, and the recoveries were 86.44-109.40% and 86.62-115.32%, respectively. The proposed fluorescent sensor provides a rapid, simple, and sensitive detection method for Cd2+ and Hg2+ in food samples and thus a novel quantitative detection method for heavy metal ions in foods.

Selection and colorimetric application of ssDNA aptamers against metamitron based on magnetic bead-SELEX.

Metamitron (MTM) is a typical and widely used triazine herbicide in agricultural production. Its moderate toxicity and high residue in the environment have deleterious impacts on human health. The establishment of a rapid and efficient MTM detection method is of great significance. In this study, a magnetic-bead SELEX (systematic evolution of ligands by exponential enrichment) system was developed to select the MTM aptamers with high affinity and specificity. Through 10 rounds of screening, six candidate aptamers with the highest abundance were obtained by high-throughput sequencing. The homology, secondary structure, and affinity analyses were performed. The aptamer named MTM-6 was selected as the optimal aptamer with the dissociation constant (Kd) value of 16 nM. Then, a colorimetric detection method for MTM based on aptamer MTM-6 and the aggregation of gold nanoparticles (AuNPs) induced by NaCl was established with a linear range from 20 to 1000 nM (R = 0.9966) and a limit of detection (LOD) of 4.58 nM. The average recovery rate of MTM in the application of actual aqueous samples ranged from 95.40 to 107.83% with a relative standard deviation (RSD) from 1.11 to 3.48%. With considerable sensitivity and specificity, this colorimetric aptasensor is convenient and efficient, and shows bright application potential in MTM detection in aqueous samples.

Fluorescent carbon dots from water hyacinth as detection sensors for ferric ions: the preparation and optimisation using response surface methodology.

The search for alternatives to chemicals from natural products as precursors for the preparation of highly doped carbon dots (CDs) remains challenging. Novel CDs (W-CDs) were synthesised using a one-step pyrolysis method with wastewater hyacinth as the sole carbon and nitrogen source at a mild temperature without using any surface-activating reagents or salt. The obtained W-CDs emitted strong blue fluorescence under 365 nm UV light excitation, with a quantum yield of 15.12%. The Box-Behnken design of the response surface methodology was applied to optimize the W-CD preparation conditions, including the reaction temperature, reaction time and weight of water hyacinths. The temperature was found to be the most important factor affecting the fluorescence intensity of the W-CDs. Additionally, the fluorescence sensor based on W-CDs demonstrated excellent selectivity towards ferric (Fe) ions, with a limit of detection of 2.35 µM. The fluorescent sensor was successfully applied for detecting Fe3+ in real water samples with a recovery of 97.80-103.10%. Hence, the pyrolysis of water hyacinth is proven to be a rapid, effective and green approach for CDs and provides a novel method for recycling water hyacinth.

Performance Analysis of an Absorption Heat Pump System for Waste Heat and Moisture Cascade Recovery from Flue Gas.

The typical flue gas exhaust temperature of coal-fired boilers is up to 150 °C. There is a considerable amount of waste heat that cannot be efficiently recovered. This study describes a zero-energy consumption absorption heat pump system that can improve the thermal efficiency and recover moisture. In the proposed system, lithium bromide solution serves as the recyclable heat-transfer medium, which absorbs sensible heat at the outlet of the air preheater by a generator. The latent heat of the flue gas is absorbed by an evaporator, and the condensate water appears. Theoretical investigation and mathematical models are built. Meanwhile, the operating parameters of the system are displayed. The results showed that the acid dew point of flue gas is related to the water content. The sorption heat pump efficiency (coefficient of performance) increased to 1.64. Compared with the two-stage heat exchanger system, 24.4 t/h condensate water and 47.21 MW waste heat were recovered. The flue gas temperature at the chimney entrance was reduced by 4.18 °C.

Selection and identification of a DNA aptamer for fluorescent detection of netilmicin.

Netilmicin (NET) is an antibiotic widely used in healthcare and agriculture, but it can accumulate in the environment to threat human health. Netilmicin (NET) is an antibiotic used for veterinary purposes, for human therapy and for agricultural purposes. Therefore, there is a need to develop high-sensitive measuring methods to detect NET. Aptamer-based detecting methods are highly sensitive, inexpensive, and portable. In this study, we developed an aptamer-based fluorescence method to detect and quantify NET. NET was first conjugated to magnetic beads by amidation reaction and then NET-coated beads were used as the stationary phase to isolate aptamers by systematic evolution of ligands by exponential enrichment (SELEX) screening method. After ten rounds of SELEX screening, 32 aptamers with NET-binding affinity were obtained and the candidate aptamer APT-21 was finally chosen by comprehensively comparing their secondary structure characters and NET-binding affinity. APT-21 bound to NET with high affinity (Kd = 194.1 nmol/L) and high specificity that it displayed low cross-binding activities on 7 different structural analogs. We also developed a fluorometric assay using SYBR Green I (SG-I) and the APT-21. Key experimental parameters were optimized, including buffer system, SG-I and APT-21 reaction time, SG-I concentration, and aptamer concentration, to improve the detecting sensitivity. Our results suggest that the low limit of detection (LOD) of this method reached a low level of 1.95 nM and it also exhibited a good linear range up to 200 nM. Moreover, we successfully applied our method to detect the NET spiked in tap water and river water with good recoveries in the range from 97% to 111%. In conclusion, our current study isolated a NET-specific aptamer and developed an aptamer-based quantification method, which is promising to apply to detect NET in environmental samples.

Experimental Investigation into Flue Gas Water and Waste Heat Recovery Using a Purge Gas Ceramic Membrane Condenser.

The direct discharge of wet saturated flue gas from a coal-fired power plant boiler causes a lot of water and waste heat loss. An inorganic ceramic membrane condenser recovers water and waste heat from the flue gas, which has great significance to improve energy utilization efficiency and reduce water consumption. However, the flue gas temperature is relatively low; thus, it is difficult to effectively utilize waste heat. In this paper, it is attempted to use the boiler secondary air as the cooling medium of the ceramic membrane condenser to realize the flue gas waste heat reuse. Based on the above ideas, a purge gas ceramic membrane condenser experimental platform was built for the water and waste heat recovery from the flue gas, and the water and waste heat recovery characteristics and the purge gas outlet parameters were discussed. Simultaneously, the heat transfer resistance and water recovery power consumption are also analyzed. The experimental results show that the water and waste heat recovery characteristics are enhanced with the purge gas flow increases. Increasing the flue gas temperature will increase the water recovery rate and heat recovery power. The ceramic membrane transmission efficiency is a key factor in restricting the actual water recovery efficiency. The purge gas absorbs the water and waste heat from the flue gas, the purge gas temperature and moisture content are significantly increased, and the purge gas relative humidity is also close to saturation. The Biot number of the ceramic membrane condenser is about 3.2 × 10-3 to 1.9 × 10-2; thus, the ceramic membrane tube wall thermal resistance can be neglected. There is a temperature difference between the flue gas and the purge gas, and the entropy production value of the ceramic membrane condenser increases with the flue gas temperature increases by the irreversible process.

Reducing environmental risks of chlorpyrifos application in typical soils by adding appropriate exogenous organic matter: Evidence from a simulated paddy field experiment.

Chlorpyrifos (CPF), as an organophosphate insecticide extensively used in the modern agricultural system, has been gradually banned in many countries due to its reported health risks to organisms, including humans. This study used simulated paddy field experiments and carbon-14 tracing to explore the possibility of reducing environmental risks of chlorpyrifos application through appropriate agronomic practice. Results showed 14C-CPF concentration in rice plants planted in the red soil (RS) was significantly higher than that in black soil (BS) and fluvo-aquic soil (FS). The application of biochar and chicken manure in RS reduced 14C-CPF accumulation in rice plants, and the content of 14C-CPF in rice grains decreased by 25% and 50%, respectively. Adding biochar to all three soils reduced the migration of 14C-CPF, especially in FS with the highest risk of 14C-CPF migration. The addition of chicken manure in FS reduced the migration of 14C-CPF and the total residual amount of 14C-CPF in the soil. In addition, chicken manure treatment increased the formation of 14C-bound residues (BRs) in soils and changed the distribution 14C-BRs in humus. The results indicated that the degree of environmental risks associated with the CPF application varies with soil types and could be reduced by introducing suitable exogenous organic matter into different soils, which is of great significance for guiding the scientific application of chlorpyrifos in agronomic practices.

Transcriptome analysis reveals the molecular mechanism of long-term exposure of Eriocheir sinensis to low concentration of trichlorfon.

Trichlorphon, a common organophosphorus pesticide (OPs), is widely used in aquaculture to prevent aquatic insects from infecting cultured objects as well as to control the excessive proliferation of plankton in water bodies. However, its repeated use time can contaminate water bodies and impart direct/indirect toxicity to beneficial aquatic species. However, the underlying mechanism regarding toxicity and cellular metabolism remains unclear. Understanding the mechanism would enable the standardized use and management of OPs and their use in the aquatic environment. Here, low concentration of trichlorphon (5 × 10-5 g/L) was used to construct a hepatopancreatic transcriptional library 30 d, 60 d and 90 d after exposure using RNA-Seq. We detected 649, 148, and 2949 DEGs in the hepatopancreas of E. sinensis for the Tri01 vs. Ctr01, Tri02 vs. Ctr02 and Tri03 vs. Ctr03 library, respectively. The results of KEGG pathway enrichment analysis showed that DEGs were mainly enriched in signal transduction, carbohydrate metabolism, transport and catabolism, endocrine system, and digestive system. Also, under trichlorfon stress, DEGs of E. sinensis were enriched in thyroid hormone signaling pathways, protein digestion and absorption, cancer pathways, etc. The significant DEGs were mainly related to metabolism and the apoptosis and autophagy pathways. This study lays a foundation for further revealing the effects of long-term trichlorfon stress on E. sinensis as well as the potential physiological toxicity. The relevant transcriptome data could provide a reference for the molecular toxicological evaluation of trichlorfon in aquaculture.