[Toxicity of bisphenol S on human normal ovarian epithelial cells mediated by ERß-MAPK signaling pathway].

OBJECTIVE: To investigate the effects of long-term(7 days and 14 days) bisphenol S(BPS) exposure on the ERß-MAPK signaling pathway, hormone secretion phenotype and cell cycle in human normal ovarian epithelial cells IOSE 80 at actual human exposure level. METHODS: Physiologically based pharmacokinetic model combined with BPS levels in the serum of women along the Yangtze River in China was used to determine the dosing concentrations of BPS, and vehicle control and 17 ß-estradiol(E_2) control were used. Complete medium with corresponding concentrations(0, 6.79×10~(-6), 6.79×10~(-4), 6.79×10~(-2), 6.79 µmol/L BPS and 10 nmol/L E_2) was replaced every 2 days. mRNA expressions of estrogen receptor(ERß and GPR30), key genes in MAPK signaling pathway(P38/JNK/ERK signaling pathway) and gonadotropin-releasing hormone-related genes(GnRH-I, GnRH-II and GnRH-R) were measured by qPCR. The ERß-MAPK signaling pathway inhibitors were employed to detect the effect of long-term exposure to BPS on the cell cycle by flow cytometry. Dose-response relationship analysis was performed to calculate the benchmark does lower confidence limits. RESULTS: Compared to the vehicle control, after 7 days exposure to BPS, the ratio of G_2/M phase was significantly increased(P<0.05), and the mRNA expressions of GnRH-I, GnRH-II and GnRH-R were significantly decreased(P<0.05); after 14 days exposure to BPS, the mRNA expressions of ESR2, MAPK3, and MAPK9 were significantly increased(P<0.05), and the mRNA expressions of GnRH-II and GnRH-R were significantly decreased(P<0.05). The GnRH-II mRNA expression level of BPS treatment for 7 days; the G_0/G_1 phase ratio, MAPK3 and MAPK8 mRNA expression level of BPS exposure for 14 days; and the GnRH-I mRNA expression level after BPS treatment for 7 days and 14 days showed a good dose-response relationship but with poor fit. CONCLUSION: Long-term low-dose exposure to BPS may cause cell cycle arrest by activating the ERß-MAPK signaling pathway, and may lead to changes in the hormone secretion of IOSE 80 cells.

FTO genotype and body mass index reduction in childhood obesity interventions: A systematic review and meta-analysis.

Numerous guidelines have called for personalized interventions to address childhood obesity. The role of fat mass and obesity-associated gene (FTO) in the risk of childhood obesity has been summarized. However, it remains unclear whether FTO could influence individual responses to obesity interventions, especially in children. To address this, we systematically reviewed 12,255 records across 10 databases/registers and included 13 lifestyle-based obesity interventions (3980 children with overweight/obesity) reporting changes in body mass index (BMI) Z-score, BMI, waist circumference, waist-to-hip ratio, and body fat percentage after interventions. These obesity-related outcomes were first compared between children carrying different FTO genotypes (rs9939609 or its proxy) and then synthesized by random-effect meta-analysis models. The results from single-group interventions showed no evidence of associations between FTO risk allele and changes in obesity-related outcomes after interventions (e.g., BMI Z-score: -0.01; 95% CI: -0.04, 0.01). The results from controlled trials showed that associations between the FTO risk allele and changes in obesity-related outcomes did not differ by intervention/control group. To conclude, the FTO risk allele might play a minor role in the response to obesity interventions among children. Future studies might pay more attention to the accumulation effect of multiple genes in the intervention process among children.

Integrated sentence-level speech perception evokes strengthened language networks and facilitates early speech development.

Natural poetic speeches (i.e., proverbs, nursery rhymes, and commercial ads) with strong prosodic regularities are easily memorized by children and the harmonious acoustic patterns are suggested to facilitate their integrated sentence processing. Do children have specific neural pathways for perceiving such poetic utterances, and does their speech development benefit from it? We recorded the task-induced hemodynamic changes of 94 children aged 2 to 12 years using functional near-infrared spectroscopy (fNIRS) while they listened to poetic and non-poetic natural sentences. Seventy-three adult as controls were recruited to investigate the developmental specificity of children group. The results indicated that poetic sentences perceiving is a highly integrated process featured by a lower brain workload in both groups. However, an early activated large-scale network was induced only in the child group, coordinated by hubs for connectivity diversity. Additionally, poetic speeches evoked activation in the phonological encoding regions in the children's group rather than adult controls which decreases with children's ages. The neural responses to poetic speeches were positively linked to children's speech communication performance, especially the fluency and semantic aspects. These results reveal children's neural sensitivity to integrated speech perception which facilitate early speech development by strengthening more sophisticated language networks and the perception-production circuit.

Mode of action exploration of reproductive toxicity induced by bisphenol S using human normal ovarian epithelial cells through ERß-MAPK signaling pathway.

BACKGROUND: In the plastics production sector, bisphenol S (BPS) has gained popularity as a replacement for bisphenol A (BPA). However, the mode of action (MOA) of female reproductive toxicity caused by BPS remains unclear and the safety of BPS is controversial. METHODS: Human normal ovarian epithelial cell line, IOSE80, were exposed to BPS at human-relevant levels for short-term exposure at 24 h or 48 h, or for long-term exposure at 28 days, either alone or together with five signaling pathway inhibitors: ICI 18,2780 (estrogen receptor [ER] antagonist), G15 (GPR30 specific inhibitor), U0126 (extracellular regulated protein kinase [ERK] 1/2 inhibitor), SP600125 (c-Jun N-terminal kinase [JNK] inhibitor) or SB203580 (p38 mitogen­activated protein kinase [p38MAPK] inhibitor). MOA through ERß-MAPK signaling pathway interruption was explored, and potential thresholds were estimated by the benchmark dose method. RESULTS: For short-term exposure, BPS exposure at human-relevant levels elevated the ESR2 and MAPK8 mRNA levels, along with the percentage of the G0/G1 phase. For long-term exposure, BPS raised the MAPK1 and EGFR mRNA levels, the ERß, p-ERK, and p-JNK protein levels, and the percentage of the G0/G1 phase, which was partly suppressed by U0126. The benchmark dose lower confidence limit (BMDL) of the percentage of the S phase after 24 h exposure was the lowest among all the BMDLs of a good fit, with BMDL5 of 9.55 µM. CONCLUSIONS: The MOA of female reproductive toxicity caused by BPS at human-relevant levels might involve: molecular initiating event (MIE)-BPS binding to ERß receptor, key event (KE)1-the interrupted expression of GnRH, KE2-the activation of JNK (for short-term exposure) and ERK pathway (for long-term exposure), KE3-cell cycle arrest (the increased percentage of the G0/G1 phase), and KE4-interruption of cell proliferation (only for short-term exposure). The BMDL of the percentage of the S phase after 24 h exposure was the lowest among all the BMDLs of a good fit, with BMDL5 of 9.55 µM.

ß-Elemene in zedoary turmeric oil injection induces dyspnea by binding to hemoglobin and upregulating HIF-1α.

ETHNOPHARMACOLOGICAL RELEVANCE: Zedoary turmeric oil injection (ZTOI) extracted from the rhizome extract of Curcuma phaeocaulis Valeton, Curcuma wenyujin Y. H. Chen et C. Ling or Curcuma kwangsiensis S. G. Lee et C. F. Liang, is widely used for the treatment of virus-induced upper respiratory tract infections, peptic ulcers, viral pneumonia, etc. However, it has attracted widespread attention because it often causes adverse drug reactions (ADRs), including dyspnea. However, little is known about the mechanism underlying dyspnea caused by ZTOI, which limits its clinical application. AIM OF THE STUDY: To investigate the major pathophysiologic signatures and underlying mechanism of ZTOI-related dyspnea. METHODS: Respiratory function detection was used to explore the pathophysiologic signature of dyspnea induced by ZTOI. UV-vis absorption spectroscopy and isothermal titration calorimetry were applied to test the interaction between ZTOI and hemoglobin (Hb). GC‒MS was used to identify the main components in ZTOI. Molecular docking, surface plasmon resonance, and circular dichroism spectroscopy were employed to test the reaction between ß-elemene and Hb. Western blot was performed to investigate the effect of ß-elemene on the hypoxia signaling pathway. RESULTS: The results showed that ZTOI-induced dyspnea was related to a decreased oxygen carrying capacity of Hb. The molecular interaction between ZTOI and Hb was proven. Notably, ß-elemene in ZTOI exhibited high binding affinity to Hb and altered its secondary structure. Furthermore, it was found that ß-elemene downregulated the expression of prolyl hydroxylase-domain protein 2 and upregulated the expression of hypoxia-inducible factor-1α. CONCLUSIONS: Our study is valuable for better understanding the pathophysiological characteristics and underlying mechanism of ZTOI to ensure its safe clinical application. We also provided a strategy to elucidate the underlying mechanism based on inspiration from clinical ADR phenotypes for investigating other medical products with ADRs in the clinic.

Comparative pharmacokinetic analysis of six major bioactive constituents using UPLC-MS/MS in samples isolated from normal and diabetic nephropathy rats after oral administration of Gushen Jiedu capsule.

Berberine, palmatine, physcion, rhein, calycosin-7-O-glucoside, and ferulic acid are six major active consituents that are present in Gushen Jiedu capsule (GSJD) extracts. The aim of this study was to determine the pharmacokinetics of the six active consituents in vivo by a rapid, sensitive, and precise UPLC-MS/MS method, which were compared between normal and diabetic nephropathy (DN) rats. Good separation of the target analytes and internal standards (ketoprofen and puerarin) was obtained on a Waters BEH C18 UPLC column with a mobile phase of 0.1 % formic acid acetonitrile-0.1 % formic acid water. All the calibration curves showed good linearity with a regression coefficient (r2) of ≥ 0.9908. The lower limits of quantification (LLOQ) for berberine, palmatine, physcion, rhein, calycosin-7-O-glucoside, and ferulic acid were 20, 2.5, 20, 20, 2.5, and 2.5 ng/mL, respectively. The relative standard deviations (RSDs) of intra-day and inter-day precision were all within 12.66 %, and the relative errors of intra-day and inter-day accuracy ranged from - 15.00 to 14.93 %. Good extraction recovery and matrix effects were obtained. The stability study confirmed the stability of the six analytes (RSD < 15 %). Finally, the data showed that the pharmacokinetic parameters (especially CLz/F, AUC and Tmax) of the six target analytes in DN rats were significantly different from those in normal rats. PK studies under pathological conditions could provide new thoughts to elucidate the underlying mechanism of GSJD and promote the clinical development of GSJD to treat DN.

Early Visual Deprivation Impairs Functional Development of the Visual Ventral Stream.

Purpose: To probe the dynamic alternations of neural networks in real-time visual processing after visual deprivation (VD) removal. Methods: A prospective cross-sectional study was conducted. Twenty children with a history of early binocular VD caused by congenital cataracts and 20 matched typically developing (TD) children were enrolled. The event-related potential (ERP) data were obtained via high-density electroencephalography. ERP data were analyzed based on three components (P1, N170, and P2), three test conditions (objects, human faces, and Chinese characters), and peak time and region of interest (ROI) chosen on a grand average head map collapsed from the averaged waveform of each group. Source localization and alpha power spectrum density were applied to define the functional pattern of brain areas and evaluate the attention function. Results: The VD group showed significantly lower P1 amplitudes than the TD group under all conditions in peak ROIs, which were situated in the left occipito-temporal region. For both VD and TD groups, there were strong N170 effects in the character and human face conditions in the component's peak ROIs. Furthermore, source mapping indicated that the VD group generally showed significantly lower activation in the visual cortex and ventral stream, whereas the beyond network areas (mostly frontal areas) intensively participated in functional compensation in the VD group. The VD group showed significant poststimulus alpha desynchronization in object recognition. Conclusions: Our research described the mechanisms of visual networks after early binocular VD removal. Our findings may provide a new basis for the poor visual recovery after early binocular VD removal and offer clues for visual recovery strategies.

Genetic and molecular exploration of maize environmental stress resilience: Toward sustainable agriculture.

Global climate change exacerbates the effects of environmental stressors, such as drought, flooding, extreme temperatures, salinity, and alkalinity, on crop growth and grain yield, threatening the sustainability of the food supply. Maize (Zea mays) is one of the most widely cultivated crops and the most abundant grain crop in production worldwide. However, the stability of maize yield is highly dependent on environmental conditions. Recently, great progress has been made in understanding the molecular mechanisms underlying maize responses to environmental stresses and in developing stress-resilient varieties due to advances in high-throughput sequencing technologies, multi-omics analysis platforms, and automated phenotyping facilities. In this review, we summarize recent advances in dissecting the genetic factors and networks that contribute to maize abiotic stress tolerance through diverse strategies. We also discuss future challenges and opportunities for the development of climate-resilient maize varieties.

A missense mutation in RRM1 contributes to animal tameness.

The increased tameness to reduce avoidance of human in wild animals has been long proposed as the key step of animal domestication. The tameness is a complex behavior trait and largely determined by genetic factors. However, the underlying genetic mutations remain vague and how they influence the animal behaviors is yet to be explored. Behavior tests of a wild-domestic hybrid goat population indicate the locus under strongest artificial selection during domestication may exert a huge effect on the flight distance. Within this locus, only one missense mutation RRM1I241V which was present in the early domestic goat ~6500 years ago. Genome editing of RRM1I241V in mice showed increased tameness and sociability and reduced anxiety. These behavioral changes induced by RRM1I241V were modulated by the alternation of activity of glutamatergic synapse and some other synapse-related pathways. This study established a link between RRM1I241V and tameness, demonstrating that the complex behavioral change can be achieved by mutations under strong selection during animal domestication.

Monotropein: A comprehensive review of biosynthesis, physicochemical properties, pharmacokinetics, and pharmacology.

Monotropein, a principal natural compound in iridoid glycosides extracted from Morindae officinalis radix, has potent pharmacological activities. To understand and utilize monotropein, we systematically summarized the studies on monotropein, including its biosynthetic pathway, physicochemical properties, pharmacokinetics, and pharmacology. Interestingly, we found that the multiple bioactivities of monotropein, such as anti-osteoporosis, anti-inflammation, anti-oxidation, anti-nociception, and hepatic or renal protection, are closely associated with its capability of downregulating the nuclear factor-κB signaling pathway, inhibiting the mitogen-activated protein kinase signaling pathway, attenuating the activation of nuclear factor E2-related factor 2/heme oxygenase-1 signaling pathway, and regulating the mammalian target of rapamycin/autophagy signaling pathway. However, the clinically therapeutic effects and the potential problems need to be addressed. This review highlights the current research progress on monotropein, which provides a reference for further investigation of monotropein.

Genome assembly and genetic dissection of a prominent drought-resistant maize germplasm.

In the context of climate change, drought is one of the most limiting factors that influence crop production. Maize, as a major crop, is highly vulnerable to water deficit, which causes significant yield loss. Thus, identification and utilization of drought-resistant germplasm are crucial for the genetic improvement of the trait. Here we report on a high-quality genome assembly of a prominent drought-resistant genotype, CIMBL55. Genomic and genetic variation analyses revealed that 65 favorable alleles of 108 previously identified drought-resistant candidate genes were found in CIMBL55, which may constitute the genetic basis for its excellent drought resistance. Notably, ZmRtn16, encoding a reticulon-like protein, was found to contribute to drought resistance by facilitating the vacuole H+-ATPase activity, which highlights the role of vacuole proton pumps in maize drought resistance. The assembled CIMBL55 genome provided a basis for genetic dissection and improvement of plant drought resistance, in support of global food security.

The battle of crops against drought: Genetic dissection and improvement.

With ongoing global climate change, water scarcity-induced drought stress remains a major threat to agricultural productivity. Plants undergo a series of physiological and morphological changes to cope with drought stress, including stomatal closure to reduce transpiration and changes in root architecture to optimize water uptake. Combined phenotypic and multi-omics studies have recently identified a number of drought-related genetic resources in different crop species. The functional dissection of these genes using molecular techniques has enriched our understanding of drought responses in crops and has provided genetic targets for enhancing resistance to drought. Here, we review recent advances in the cloning and functional analysis of drought resistance genes and the development of technologies to mitigate the threat of drought to crop production.

Profiles of total and sn-2 fatty acid of human mature milk and their correlated factors: A cross-sectional study in China.

Fatty acid (FA) in breast milk is beneficial to the growth and neurodevelopment of infants. However, the structure profiles of breast milk FAs and the influencing factors which are crucial for normal function have not been fully elucidated. This study aimed to characterize the profiles of total and sn-2 FAs in human mature milk based on two representative urban areas in China and explore potential sociodemographic determinants. Mothers (n = 70) at 40-100 d postpartum from Beijing and Danyang were recruited according to unified inclusion and exclusion criteria. Total and sn-2 FA compositions were examined by gas chromatography and quantified. Using the Spearman correlation and multiple regression model, we found that the location and maternal education level were the most conspicuous correlated factor. The milk of mothers from Beijing had higher levels of the n-6 series of long-chain polyunsaturated fatty acids (LCPUFA) (C20:2, C20:3n-6, C20:4n-6, n-6PUFA/n-3PUFA, LA/ALA, and ARA/DHA) than that of Danyang, while the opposite was observed in the n-3 series of LCPUFA (C18:3n-3 and Total n-3PUFA). Compared to the milk of mothers with a high school degree or below, those with a bachelor's degree or above had lower SFAs (C10:0, C12:0, C14:0, and Total SFA), n-3 series of LCPUFA (C18:3n-3 and Total n-3PUFA), C18:1n-9t, and higher n-6 series of LCPUFA (C18:2n-6c, C20:2, C20:4n-6, Total n-6PUFA, and n-6PUFA/n-3PUFA). Maternal age, infant gender, pre-conception body mass index (BMI), parity, delivery mode, and gestational weight gain were also associated with total FAs. However, fewer associations were found between the above factors and sn-2 FAs. This study will promote an understanding of human breast milk's lipid profile and help develop a formula more suitable for infants.

The key micronutrient copper orchestrates broad-spectrum virus resistance in rice.

Copper is a critical regulator of plant growth and development. However, the mechanisms by which copper responds to virus invasion are unclear. We previously showed that SPL9-mediated transcriptional activation of miR528 adds a previously unidentified regulatory layer to the established ARGONAUTE (AGO18)-miR528-L-ascorbate oxidase (AO) antiviral defense. Here, we report that rice promotes copper accumulation in shoots by inducing copper transporter genes, including HMA5 and COPT, to counteract viral infection. Copper suppresses the transcriptional activation of miR528 by inhibiting the protein level of SPL9, thus alleviating miR528-mediated cleavage of AO transcripts to strengthen the antiviral response. Loss-of-function mutations in HMA5, COPT1, and COPT5 caused a significant reduction in copper accumulation and plant viral resistance because of the increased SPL9-mediated miR528 transcription. Gain in viral susceptibility was mitigated when SPL9 was mutated in the hma5 mutant background. Our study elucidates the molecular mechanisms and regulatory networks of copper homeostasis and the SPL9-miR528-AO antiviral pathway.

Polystyrene microplastic exposure induces insulin resistance in mice via dysbacteriosis and pro-inflammation.

Microplastics (MPs) as emerging contaminants have become a global environmental problem. However, studies on the effects of MPs on metabolic diseases remain limited. Here, we evaluated the effects of polystyrene (PS), one of the most prominent types of MPs, on insulin sensitivity in mice fed with normal chow diet (NCD) or high-fat diet (HFD), and explained the underlying mechanisms. Mice fed with NCD or HFD both showed insulin resistance (IR) after PS exposure accompanied by increased plasma lipopolysaccharide and pro-inflammatory cytokines such as tumor necrosis factor-α and interleukin-1ß. Exposure to PS also resulted in a significant decrease in the richness and diversity of gut microbiota, particularly an increase in the relative abundance of Gram-negative bacteria such as Prevotellaceae and Enterobacteriaceae. Additionally, PS with a small particle size (5 µm) accumulated in the liver, kidneys and blood vessels of mice. Further analyses showed inhibition of the insulin signaling pathway in the liver of PS exposed mice, such as inhibition of IRS1 and decreased expression of PI3K. Hence, the mechanism of PS exposure to induce IR in mice might be mediated through regulating gut microbiota and PS accumulation in tissues, stimulating inflammation and inhibiting the insulin signaling pathway. In conclusion, PS might be a potential environmental contaminant that causes metabolic diseases associated with IR.

The level and efficacy of lutein in patients with age-related macular degeneration: a comprehensive systematic review and meta-analysis.

Background: Lutein has been linked with various visual performance disorders, including age-related macular degeneration (AMD). However, previous studies evaluating the association between serum lutein and the risk of AMD showed results, and the efficacy of lutein intake in AMD patients remains unclear. Methods: To comprehensively estimate the relationship between lutein and AMD, a systematic review and meta-analysis was conducted by searching eligible randomized clinical trials (RCT) and case-control studies to study the association between lutein and AMD on the Cochrane Library, MEDLINE, Elsevier, PubMed, Web of Knowledge, Chinese National Knowledge Infrastructure (CNKI), and Chinese Biomedical Database (CBM) databases until April 2020. The weighted mean difference (WMD) with 95% confidence interval (95% CI) was adopted as the primary effect estimate. Meta-analysis was conducted using STATA 12.0. Results: Nine studies with 855 participants were included in this meta-analysis. The NOS scoring of five case-control studies ranged 5-9. For RCTs, two studies were rated with a low risk of bias, one study with a moderate risk of bias and one with a high risk of bias. The results increased significantly in macular pigment optical density (MPOD) (WMD =0.069; 95% CI: 0.040-0.098, P=0.000) among AMD patients taking lutein supplementation, while there was no difference in circulating lutein levels between AMD patients and controls (WMD =0.00; 95% CI: -0.01 to 0.00, P=0.310). Subgroup analysis suggested that the dose and duration of supplementation could significantly influence the MPOD level in AMD patients. In particular, we observed a larger increase in MPOD of AMD patients using a higher dose (20 mg/d) and longer treatment (>6 months). Conclusions: Although current evidence does not support circulating lutein as a biomarker for early screening of the high-risk AMD population, this study is the first meta-analysis to explore the relationship between lutein in blood and AMD patients. Given that lutein has a high safety profile as indicated by many studies, it is reasonable to give the current analysis result that high dose (20 mg/d) and long duration (>6 months) of lutein intake could be beneficial to AMD patients.

Transcriptomics analysis and benchmark concentration estimating-based in vitro test with IOSE80 cells to unveil the mode of action for female reproductive toxicity of bisphenol A at human-relevant levels.

Bisphenol A (BPA) is of great concern in public health, of which female reproductive toxicity is one major adverse health effect with the unclear mode of action (MOA) yet. Based on the principle of Toxicity Testing in the 21st Century, the purpose of this study is to explore the MOA for female reproductive toxicity using human normal ovarian epithelial cells IOSE80 at 28-day human-relevant-level exposure. A physiological based pharmacokinetic model was used to select the administration concentrations according to the BPA levels in female gonads at human actual exposure scenario. Enrichment KEGG pathways interrupted by BPA consisted of RNA transport, ribosome biogenesis in eukaryotes, cell cycle, cellular senescence, progesterone-mediated oocyte maturation, and oocyte meiosis. Increased relative mRNA and protein expressions of ERK and CDKN3, and proportion of S phase, as well as decreased proportion of G0/G1 phase were observed with increasing BPA concentrations, which could be partially inhibited by ERK inhibitor U0126. Among all the benchmark concentration lower confidence limits, mRNA expression of MAPK3 served as the lowest. In conclusion, the MOA of BPA induced female reproductive toxicity at human-relevant levels may include: key event (KE)1-ERK activation, KE2-increased expression of CDKN3, and KE3-cell cycle arrest. However, more in vivo studies may be needed to complete the MOA.

Network pharmacology-based dissection of the underlying mechanisms of dyspnoea induced by zedoary turmeric oil.

Zedoary turmeric oil (ZTO) has been widely used in clinic. However, the unpleasant induced dyspnoea inevitably impedes its clinical application. Thus, it is urgent to elucidate the mechanism underlying the ZTO-induced dyspnoea. In this study, network pharmacology was firstly performed to search the clue of ZTO-induced dyspnoea. The key target genes of ZTO-induced dyspnoea were analysed using GO enrichment analysis and KEGG pathway analysis. GO analysis suggested that haem binding could be a key molecular function involved in ZTO-induced dyspnoea. Hence, the haemoglobin (Hb) was focused for its oxygen-carrying capacity with haem as its critical component binding to the oxygen. Ultraviolet-visible absorption spectrum indicated that the ZTO injection (ZTOI) perturbed the Soret band of Hb, suggesting an interaction between ZTO and Hb. GC-MS analysis revealed that ß-elemene, germacrone, curdione and furanodiene were main components of ZTOI. Molecular docking was used to illustrate the high affinity between representative sesquiterpenes and Hb, which was finally confirmed by surface plasmon resonance, suggesting their potential roles in dyspnoea by ZTO. Following a network pharmacology-driven strategy, our study revealed an intervened Hb-based mechanism underlying the ZTO-induced dyspnoea, providing a reference for elucidating mechanism underlying adverse drug reactions of herbal medicines in clinic.