Abstract. Ground-level ozone (O3) pollution in river–valley cities is influenced by interactions among photochemistry, boundary-layer processes, and terrain-induced circulations, yet their combined effects on O3 distributions remain insufficiently understood. Using WRF–CAMx coupled with source apportionment and process analysis, we investigated the three-dimensional distribution, source contributions, and formation characteristics of O3 over the Nanjing section of the Yangtze River. The altitude of the O3 maximum decreased from ~5.0 km in winter to ~2.0 km in summer, indicating stronger near-surface photochemical production and boundary-layer processes during warm seasons. Distinct spatial heterogeneity was observed between the river corridor and adjacent urban areas. Daytime near-surface O3 development was weaker over the river in summer, whereas the high-O3 layer extended downward to greater depths in spring and autumn. Regional background transport accounted for most total O3, while suburban contributions increased during high-O3 episodes, reaching 45.5 % under O3 concentrations of at least 160 µg/m3 in July. Transport and diffusion associated with river-breeze circulations played important roles in regulating O3 variability within the river corridor. O3 formation remained volatile organic compound (VOC)-limited in spring and autumn, whereas river-breeze-induced nitrogen oxide (NOx) dilution shifted the river corridor to NOx-limited conditions approximately two hours earlier than in the surrounding urban area in summer. These findings demonstrate that river-induced circulations can substantially modify O3 distributions, source contributions, and chemical sensitivity within river–valley cities, and that city-averaged and monthly mean characterizations may not fully capture local O3 variability in complex terrain.

Chimeric antigen receptor-engineered macrophages (CAR-Ms) hold great promise for solid tumor immunotherapy. The intracellular domains (ICDs) of CARs determine the phenotypic output of therapeutic macrophages but remain largely unexplored. Here, we constructed a CAR library containing 131 unique signaling domains derived from native immune receptors and identified 17 ICDs that enhance macrophage phagocytosis, inflammatory responses, or tumor infiltration in vitro and in vivo. We further developed a scalable 3′ barcode technology, CARode, to uniquely label and trace ICD variants within large-scale combinatorial CAR library and applied it to single-cell RNA sequencing and single-cell CAR analysis to assess the synergetic effects of ICD combinations on macrophage activation. Our approach uncovered a novel CD40-LY9-FCRL1 chimeric receptor that modulates the tumor microenvironment and improves solid tumor clearance. In conclusion, our findings demonstrate that pooled screening can accelerate the discovery of complex ICD constructs, providing a powerful platform for engineering macrophage-based immunotherapies. ### Competing Interest Statement The Shen lab has received research support from Macera therapeutics. P. Shen is a compensated co-founder, member of the scientific advisory board, and works as the CSO of Macera therapeutics. Y. Chen is a compensated co-founder, and works as the CEO of Macera therapeutics. W. Zhang works as the Process Development Director of Macera therapeutics. P. Shen, Y. Wang, S. Zuo, Y. Chen, and W. Zhang are listed on patent applications related to this work. The other authors declare no competing interests.

ABSTRACT:Mesenchymal stem cells (MSCs) are a class of protocells that can differentiate into various cell types and have robust replication and renewal capabilities. MSCs secrete various nutritional factors to regulate the microenvironment of tumor tissues. The mechanism by which they inhibit or promote tumor growth may be closely related to MSC-derived exosomes (MSC-Exo). However, the role of MSC-Exo vesicles in tumors remains controversial. This review discusses the potential applications of microRNAs in exosomes derived from MSCs in treating tumors.

The frequent detection of emerging organic pollutants in the surface water of the Yangtze River poses a threat to ecological health, and differences exist in ecological risks in different regions, which are related to the socio-economic development of cities along the Yangtze River. To comprehensively analyze the pollution characteristics of new pollutants in the surface waters of the Yangtze River and the key factors affecting their ecological risks, this review study analyzed the data on pollutants in the Yangtze River Region from the published literature over the past 20 years. First, we used the ecological risk assessment strategy to screen the polluted areas that require attention and prioritized new pollutants, followed by revealing the links between urban social development and pollutant occurrence. The results showed that the new pollutants in the surface water of the Yangtze River mainstem included three categories, 20 subcategories, and 412 types, and the detected concentrations ranged from 10-5 to 103 ng·L-1, among which the concentrations of volatile organic pollutants （VOPs）, phthalate esters （PAEs）, insecticides, fungicides, and herbicides were high. Significant differences were observed in the concentration and types of surface water pollutants among different regions, namely more in the east and less in the west, with industrial categories concentrated in the downstream （Anhui-Shanghai） and pesticides concentrated in the midstream （Wuhan-Anhui）. Most emerging pollutants with high ecological risk belonged to the industrial product category and the pharmaceutical and personal care product category. The spatial distribution characteristics of pollutants were greatly influenced by population density, urban area, per capita GDP, and the proportion of secondary industry, and the degree of human activities in cities was the main reason for the large differences in the distribution of pollutants.

Exposed to ubiquitously perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) has been associated with non-alcoholic fatty liver disease (NAFLD), yet the underlying molecular mechanism remains elusive. The extrapolation of empirical studies correlating per- and polyfluoroalkyl substance (PFAS) exposure with NAFLD occurrence to real-life exposure was hindered by the limited availability of mechanistic data at environmentally relevant concentrations. Herein, a novel pathway mediating hepatocyte lipid accumulation by PFOA and PFOS at human-relevant dose (<10 mu M) was identified by integrating CRISPR-Cas9 genome screening, concentration-dependent transcriptional assay in HepG2 cell and epidemiological data mining. 1) At genetic level, nudt7 showed the highest enriched potency among 569 NAFLD-related genes, and the transcription of nudt7 was significantly downregulated by PFOA and PFOS exposure (<7 mu M). 2) At molecular pathway, upon exposure to <= 10(-4) mu M PFOA and PFOS, the downregulation of nudt7 transcriptional expression triggered the reduction of Ace-CoA hydrolase activity. 3) At cellular level, increased lipids were measured in HepG2 cells with PFOA and PFOS (<2 mu M). Overall, we identified a novel mechanism mediated by transcriptional downregulation of nudt7 gene in hepatocellular lipid increase treated with PFOA and PFOS, which could potentially explain the NAFLD occurrence associated with exposure to PFASs in humans.

Macrophages may acquire a reparative phenotype that supports tissue repair and remodeling in response to tissue injury. However, the metabolic requirements underpinning this process are incompletely understood. Here, we show that posttranslational modification (PTM) of PPARγ regulates lipid synthesis in response to wound microenvironmental cues and that metabolic rewiring orchestrates function of reparative macrophages. In injured tissues, repair signaling leads to decreased macrophage PPARγ threonine 166 (T166) phosphorylation, which results in a partially active PPARγ transcriptional program comprised of increased binding activity to the regulator regions of lipid synthesis-associated genes, thereby increased lipogenesis. The accumulated lipids serve as signaling molecules, triggering STAT3-mediated growth factor expression, and supporting the synthesis of phospholipids for the expansion of the endoplasmic reticulum (ER), which is required for protein secretion. Genetic or pharmacological inhibition of PPARγ T166 phosphorylation promotes the reparative function of macrophages and facilitates tissue regeneration. In summary, our work identifies PPARγ T166-regulated lipid biosynthesis as an essential pathway for meeting the anabolic demands of the activation and function of macrophages and provides a rationale for potential therapeutic targeting of tissue repair. Macrophages with a reparative phenotype are important for tissue repair and have distinctive metabolic features. Here authors show that metabolic rewiring of macrophages during wound healing involves dephosphorylation of the transcription factor PPARγ, which results in activation of target genes that regulate lipid biosynthesis.

Herpes simplex virus type 1 (HSV-1) is a common virus infecting the ocular tissue. It infects eye tissues, such as the eyelid, cornea, and conjunctiva. Corneal HSV-1 infection causes herpes simplex keratitis (HSK), which can induce vision loss. Current treatments for eye infections targeting HSV-1 can led to various sequelae. Antiviral drugs only work during active viral replication, and viral resistance has been recorded in numerous cases. Therefore, it is necessary to determine the molecular mechanisms underlying HSV-1 infection and identify new antiviral drugs. There are no reports on whether PI3K can regulate SGK1 to modulate HSV-1 infection in corneal epithelial cells (CECs), or how this mechanism works. This study found that HSV-1 levels and apoptosis increased in human corneal epithelial cells (HCECs) and BALB/c mice after HSV-1 infection. Serum and glucocorticoid-regulated protein kinase 1 (SGK1) were upregulated in HCECs and corneal tissues of BALB/c mice infected with HSV-1, as evidenced by whole-transcriptome sequencing, quantitative real-time polymerase chain reaction (RT-qPCR), and immunofluorescence staining experiments. An inhibitor of SGK1 (GSK 650394) reduced SGK1 expression, HSV-1 replication, and apoptosis in CECs, as evidenced by western blotting, flow cytometry, and in cell western blotting. The phosphatidylinositol 3'-kinase (PI3K) pathway was activated in CECs infected with HSV-1. After treatment with the PI3K inhibitor (LY294002), the expression of SGK1 and Wnt signaling pathway protein β-catenin were downregulated, and the replication of HSV-1 decreased in CECs; additionally, CECs apoptosis was reduced. HSV-1 replication causes CECs apoptosis. In HSV-1 infected CECs, SGK1 expression was upregulated by activated PI3K/SGK1 signaling pathway. Additionally, SGK1 activated Wnt/β-catenin signaling pathway to promote HSV-1 replication and cause CEC apoptosis. In conclusion, SGK1 is an important target for HSK treatment.

Recombinase polymerase amplification (RPA) running at 37-42 degrees C is fast, efficient and less-implemented; however, the existing technologies of nucleic acid testing based on RPA have some limitations in specificity of single-base recognition and multiplexing capability. Herein, we report a highly specific and multiplex RPA-based nucleic acid detection platform by combining flap endonuclease 1 (FEN1)-catalysed invasive reactions with RPA, termed as FEN1-aided RPA (FARPA). The optimal conditions enable RPA and FEN1-based fluorescence detection to occur automatically and sequentially within a 25-min turnaround time and FARPA exhibits sensitivity to 5 target molecules. Due to the ability of invasive reactions in discriminating single-base variation, this one-pot FARPA is much more specific than the Exo probe-based or CRISPR-based RPA methods. Using a universal primer pair derived from tags in reverse transcription primers, multiplex FARPA was successfully demonstrated by the 3-plex assay for the detection of SARS-CoV-2 pathogen (the ORF1ab, the N gene, and the human RNase P gene as the internal control), the 2-plex assay for the discrimination of SARS-CoV-2 wild-type from variants (Alpha, Beta, Epsilon, Delta, or Omicrons), and the 4-plex assay for the screening of arboviruses (zika virus, tick borne encephalitis virus, yellow fever virus, and chikungunya virus). We have validated multiplex FARPA with 103 nasopharyngeal swabs for SARS-CoV-2 detection. The results showed a 100% agreement with RT-qPCR assays. Moreover, a hand-held FARPA analyser was constructed for the visualized FARPA due to the switch like endpoint read-out. This FARPA is very suitable for pathogen screening and discrimination of viral variants, greatly facilitating point-of-care diagnostics.

BackgroundDiabetic retinopathy (DR) is one of the most common microvascular complications of diabetes and causes of blindness in developed countries. Our study was designed to identify immune-related genes involved in the progression of proliferative diabetic retinopathy (PDR).MethodsThe "GSE102485" dataset of neovascular membrane samples (NVMs) from type 1 and 2 diabetes mellitus patients was downloaded from the Gene Expression Omnibus database. Functional enrichment analyses, protein-protein interaction network (PPI) construction, and module analysis of immune pathways in NVMs and controls were conducted via Gene Set Enrichment Analysis and Metascape.ResultsThe significantly upregulated hallmark gene sets in DR2 and DR1 groups were involved in five immune pathways. Only CCR4, CXCR6, C3AR1, LPAR1, C5AR1, and P2RY14 were not previously reported in the context of PDR molecular pathophysiology. Except for P2RY14, all of the above were upregulated in retinal samples from experimental diabetes mouse models and human retina microvascular endothelial cells (HRMECs) treated with high glucose (HG) by quantitative Real Time Polymerase Chain Reaction (qRT-PCR).ConclusionThe genes identified herein provide insight into immune-related differential gene expression during DR progression.

AbstractBackground The activation of NLRP3 inflammasome in macrophages is a risking factor accelerating atherosclerosis (AS) progression. Here, the function of Peli1 in regulating NLRP3 inflammasome activation during AS progression were investigated. MethodsApoE−/−mice were subjected to high fat diet to construct AS modelin vivo. HE, Oil red O and Sirius red staining were adopted to analyze histopathological changes and lipid accumulation. Raw264.7 cells stimulated by ox-LDL were used as AS cellular model. Pro-inflammatory cytokines secretion was assessed using ELISA. Total m6A level was examined by m6A dot blot assay, and Peli1 m6A level was assessed using MeRIP assay. The interactions between METTL3, YTHDC2, Peli1, YB-1 and NLRP3 were analyzed by ChIP, dual-luciferase reporter gene, CoIP, RIP and/or RNA pull down assays. Results YB-1 knockdown could inhibit AS progressionin vivo, and YB-1 silencing repressed ox-LDL-mediated lipid accumulation and inflammation in macrophages by inactivating NLRP3 inflammasome. E3 ubiquitination ligase Peli1 mediated ubiquitination degradation of YB-1 during AS progression. Moreover, it was found that YTHDC2 recognized METTL3-mediated Peli1 m6A modification and mediated Peli1 mRNA degradation. Rescue studies revealed that YB-1 upregulation abrogated Peli1 upregulation’s repression on AS progression bothin vitroandin vivo. Conclusion Peli1, regulated by m6A modification, inhibited YB-1-mediated NLRP3 inflammasome activation in macrophages by promoting YB-1 ubiquitination to suppress AS progression.

New approach methodologies (NAMs), especially omics-based high-throughput bioassays have been developed rapidly, providing rich mechanistic information such as molecular initiation events (MIEs) and (sub)cellular key events (KEs) in adverse outcome pathways (AOPs). However, how to apply the knowledge of MIEs/KEs to predict adverse outcomes (AOs) induced by chemicals represents a new challenge for computational toxicology. Here, an integrated method named ScoreAOP was developed and evaluated to predict chemicals’ developmental toxicity for zebrafish embryos by integrating four related AOPs and dose-dependent reduced zebrafish transcriptome (RZT). The rules of ScoreAOP included 1) sensitivity of responsive KEs demonstrated by point of departure of KEs (PODKE), 2) evidence reliability and 3) distance between KEs and AOs. Moreover, eleven chemicals with different modes of action (MoAs) were tested to evaluate ScoreAOP. Results showed that eight of the eleven chemicals caused developmental toxicity at tested concentration in apical tests. All the tested chemicals’ developmental defects were predicted using ScoreAOP, whereas eight out of the eleven chemicals predicted by ScoreMIE which was developed to score MIEs disturbed by chemicals based on in vitro bioassays data. Finally, in terms of mechanism explanation, ScoreAOP clustered chemicals with different MoAs while ScoreMIE failed, and ScoreAOP revealed the activation of aryl hydrocarbon receptor (AhR) plays a significant role in dysfunction of cardiovascular system, resulting in zebrafish developmental defects and mortality. In conclusion, ScoreAOP represents a promising approach to apply mechanism information obtained from omics to predict AOs induced by chemicals.

Understanding the toxicological mechanisms of chemicals is essential for accurate assessments of environmental health risks. Inflammation could play a critical role in the adverse health outcomes caused by genotoxicants; however, the toxicological mechanisms underlying genotoxicants-induced inflammatory response are still limited. Here, functional genomics CRISPR screens were performed to enhance the mechanistic understanding of the genotoxicants-induced inflammatory response at low doses exposure. Key genes and pathways associated with the activities of immune cells and the production of cytokines were identified by CRISPR screens of 6 model genotoxicants. Gene network analysis revealed that three genes (TLR10, HCAR2 and TRIM6) were involved in the regulation of neutrophil apoptosis and cytokine release, and TLR10 shared a similar functional pattern with HCAR2 and TRIM6. Furthermore, adverse outcome pathway (AOP) network analysis revealed that TLR10 was involved in the molecular initiating events (MIEs) or key events (KEs) in the inflammatory response AOPs of all the 6 genotoxicants, which provided mechanistic links between TLR10 and genotoxicants-induced inflammation and respiratory diseases. Finally, functional validation tests demonstrated that TLR10 exhibited inhibitory effects on genotoxicants-induced inflammatory responses in both epithelial and immune cells. This study highlights the powerful utility of the integration of CRISPR screen and AOP network analysis in illuminating the toxicological causal mechanisms of environmental chemicals.

Determining dose-response relationship is essential for comprehensively revealing chemical-caused effects on organisms. However, uncertainty and complexity of gene/protein interactions cause the inability of traditional toxicogenomic methods (e.g., transcriptomics, proteomics and metabolomics) to effectively establish the direct relationship between chemical exposure and genes. In this work, we built an effective dose-dependent yeast functional genomics approach, which can clearly identify the direct gene-chemical link in the process of cadmium (Cd) toxification from a genome-wide scale with wide range concentrations (0.83, 2.49, 7.48, 22.45, 67.34, 202.03 and 606.1 μM). Firstly, we identified 220 responsive strains, and found that 142, 110, 91, 34, 8, 0 and 0 responsive strains can be respectively modulated by seven different Cd exposure concentrations ranging from high to low. Secondly, our results demonstrated that these genes induced by the high Cd exposure were mainly enriched in the process of cell autophagy, but ones caused by the low Cd exposure were primarily involved in oxidative stress. Thirdly, we found that the top-ranked GO biological processes with the lowest point of departure (POD) were transmembrane transporter complex and mitochondrial respiratory chain complex III, suggesting that mitochondrion might be the toxicity target of Cd. Similarly, nucleotide excision repair was ranked first in KEGG pathway with the least POD, indicating that this dose-dependent functional genomics approach can effectively detect the molecular initiating event (MIE) of cadmium toxification. Fourthly, we identified four key mutant strains (RIP1, QCR8, CYT1 and QCR2) as biomarkers for Cd exposure. Finally, the dose-dependent functional genomics approach also performed well in identifying MIE for additional genotoxicity chemical 4-nitroquinoline-1-oxide (4-NQO) data. Overall, our study developed a dose-dependent functional genomics approach, which is powerful to delve into the MIE of chemical toxification and is beneficial for guiding further chemical risk assessment.

Exosome microRNAs (miRNAs) have great research outlook in clinical therapy and biomarkers, they have been found to have a close to multiple diseases. A growing number of studies have attempted to alleviate or treat diseases through exosomes. It indicates that miRNAs in exosomes have great significance in preventing and controlling diseases in clinical research. We summarise these studies below to better understand their implications.We screened and analyzed more than 100 articles from PubMed, Web of Science, and other databases from 1987 to 2022. Data of the clinical trials are collected from clinicaltrials.gov.In this review, we introduce the source, type, and characteristics of several exosomes, summarising current research on their role in cardiovascular, nervous system disease, tumour, and other diseases. Further, we discuss their mechanism of action and future directions for development of treatments in several diseases, and highlight the significant research value and potential use of exosomes in clinical diagnosis and treatment.An increasing number of researchers have begun to explore the link between exosomal miRNAs and diseases. More exosome therapeutics will be used in future clinical trials, which may bring new hope for the diagnosis and treatment of several diseases.KEY MESSAGESExosomes have unique advantages in molecular transport and cell signal transduction.miRNAs play an essential role in the formation of multiple diseases.Research on the clinical application and potential value of exosomes is growing.

Although previous studies have confirmed the association between phthalate esters (PAEs) exposure and endocrine disorders in humans, few studies to date have systematically assessed the threats of new PAE alternatives to endocrine disruptions. Herein, zebrafish embryos were continuously exposed to two PAEs [di-n-butyl phthalate (DBP) and diisobutyl phthalate (DiBP)], two structurally related alternatives [diiononyl phthalate (DINP) and diisononyl hexahydrophthalate (DINCH)], and two non-PAE substitutes [dipropylene glycol dibenzoate (DGD) and glyceryl triacetate (GTA)], and the endocrine-disrupting effects were investigated during the early stages (8-48 hpf). For five endogenous hormones, including progesterone, testosterone, 17β-estradiol, triiodothyronine (T3), and cortisol, the tested chemicals disturbed the contents of at least one hormone at environmentally relevant concentrations (≤3.9 μM), except DINCH and GTA. Then, the concentration-dependent reduced zebrafish transcriptome analysis was performed. Thyroid hormone (TH)- and androgen/estrogen-regulated adverse outcome pathways (AOPs) were the two types of biological pathways most sensitive to PAE exposure. Notably, six compounds disrupted four TH-mediated AOPs, from the inhibition of deiodinases (molecular initiating event, MIE), a decrease in T3 levels (key event, KE), to mortality (adverse outcome, AO) with the quantitatively linear relationships between MIE-KE (|r| = 0.96, p = 0.002), KE-AO (|r| = 0.88, p = 0.02), and MIE-AO (|r| = 0.89, p = 0.02). Multiple structural analyses showed that benzoic acid is the critical toxicogenic fragment. Our data will facilitate the screening and development of green alternatives.

Since a large number of contaminants are detected in source waters (SWs) and tap waters (TWs), it is important to perform a comprehensive effect evaluation and key contributor identification. A reduced human transcriptome (RHT)-based effect-directed analysis, which consisted of a concentration-dependent RHT to reveal the comprehensive effects and noteworthy pathways and systematic identification of key contributors based on the interactions between compounds and pathway effects, was developed and applied to typical SWs and TWs along the Yangtze River. By RHT, 42% more differentially expressed genes and 33% more pathways were identified in the middle and lower reaches, indicating heavier pollution. Hormone and immune pathways were prioritized based on the detection frequency, sensitivity, and removal efficiency, among which the estrogen receptor pathway was the most noteworthy. Consistent with RHT, estrogenic effects were widespread along the Yangtze River based on in vitro evaluations. Furthermore, 38 of 100 targets, 39 pathway-related suspects, and 16 estrogenic nontargets were systematically identified. Among them, diethylstilbestrol was the dominant contributor, with the estradiol equivalent (EEQ) significantly correlated with EEQwater. In addition, zearalenone and niclosamide explained up to 54% of the EEQwater. The RHT-based EDA method could support the effect evaluation, contributor identification, and risk management of micropolluted waters.

Freshwater ecosystems are among the most threatened ecosystems on Earth. The freshwater biodiversity crisis has caused widespread global concern. Drought as one of the factors causing freshwater biodiversity is still poorly understood. Crayfish is often used in academic research as a biological indicator. In this study, flow cytometry, hematoxylin-eosin staining, and untargeted metabolomics were used to analyze the immune function, histopathology, and metabolism of crayfish under drought conditions. After drought exposure, the total hemocytes count (THC) was significantly decreased (from 8.9 × 105 mL−1 in the control group to 2.2 × 105 mL−1 at day 5). Phagocytosis decreased by 66% after 5 days of drought. The level of reactive oxygen species (ROS) in the hepatopancreas was upregulated. Moreover, histological disorder and metabolism changes in the hepatopancreas were obvious. These results indicate that drought suppresses immune function, disrupts the balance of oxidative and antioxidative systems, and induces tissue damage and metabolic changes in crayfish.

ObjectivesRegular sports participation is a gendered phenomenon in China. Women have reported much higher constraints than men on time, partner, psychology, knowledge, and interest. This study explores personal, family, lifestyle, and health factors associated with sports participation. Study DesignThis study is a cross-sectional study. MethodsData were collected from the national reprehensive China Family Panel Studies (CFPS) database (2018) to analyze personal information, family background, lifestyle, and health in relation to women's sports participation. Multiple classification logistic regression was used to quantify the association between independent variables and sports time. ResultsWomen with high personal income and education, who were unmarried, in faster economic development areas have more awareness and more time for sports participation. Women who were overweight and self-rated as unattractive spent less time on sports participation. Women with a small family population and no children have more time for sports participation. Less time on the internet and moderate sleep contribute to active sports participation. Women with chronic diseases and high medical costs are less likely to participate in sports. ConclusionsNegative body aesthetic perception, the burden of family environment, modernization of lifestyle, and the normalization of sub-health are essential factors affecting women's sports participation. The government should understand the inner and outer barriers to women's participation in sports, develop policies and regulations to protect and support women's sports participation, and guide and monitor the effective implementation of women's sports activities.

Cytotoxicity of non-polar narcotic chemicals can be predicted by quantitative structure activity relationship (QSAR) models, but the polar narcotic chemicals' actual cytotoxicity exceeds the predicted values by their chemical structures. This discrepancy indicates that the molecular mechanism by which polar narcotic chemicals exert their toxicity is unclear. Taking advantage of Saccharomyces cerevisiae (yeast) functional genome-wide heterozygous essential gene knockout mutants, we here have identified the specific molecular fingerprints of two main chemical structure groups (phenols and anilines) of polar narcotic chemicals (dichlorophen (DCP), 4-chlorophenol (4-CP), 2, 4, 6-trichlorophenol (TCP), 3, 4-dichloroaniline (DCA) and N-methylaniline (NMA)) and one non-polar narcotic chemical 2, 2, 2-trichloroethanol (TCE). Especially, we identify 33, 57, 54, 46, 59 and 53 responsive strains through exposure to TCE, DCP, 4-CP, TCP, DCA and NMA with three test concentrations, respectively, revealing that these polar narcotic chemicals have more responsive strains than the non-polar narcotic chemical. Remarkably, we find that the molecular fingerprints of polar narcotic chemicals in different chemical structure groups are obviously varied, particularly phenols and anilines have their own specific molecular fingerprints. Interestingly, our results demonstrate that the molecular toxicity mechanisms of anilines are associated with DNA replication, but phenols are related with pathway of RNA degradation. Additionally, we find that the two knockout strains (SME1 and DIS3) and the three knockout strains (TSC11, RSP5 and HSF1) can specifically respond to exposure to phenols and anilines, respectively. Thus, they may be served as potential biomarkers to distinguish phenols from anilines. Collectively, our works demonstrate that the functional genomic platform of yeast essential gene mutants can not only act as an effective tool to identify key specific molecular fingerprints for polar narcotic chemicals, but also help to understand the molecular mechanisms of polar narcotic chemicals.