Synthetic protein circuits for programmable control of mammalian cell death.

Natural cell death pathways such as apoptosis and pyroptosis play dual roles: they eliminate harmful cells and modulate the immune system by dampening or stimulating inflammation. Synthetic protein circuits capable of triggering specific death programs in target cells could similarly remove harmful cells while appropriately modulating immune responses. However, cells actively influence their death modes in response to natural signals, making it challenging to control death modes. Here, we introduce naturally inspired "synpoptosis" circuits that proteolytically regulate engineered executioner proteins and mammalian cell death. These circuits direct cell death modes, respond to combinations of protease inputs, and selectively eliminate target cells. Furthermore, synpoptosis circuits can be transmitted intercellularly, offering a foundation for engineering synthetic killer cells that induce desired death programs in target cells without self-destruction. Together, these results lay the groundwork for programmable control of mammalian cell death.

Single cell analyses of cancer cells identified two regulatorily and functionally distinct categories in differentially expressed genes among tumor subclones.

To explore the feature of cancer cells and tumor subclones, we analyzed 101,065 single-cell transcriptomes from 12 colorectal cancer (CRC) patients and 92 single cell genomes from one of these patients. We found cancer cells, endothelial cells and stromal cells in tumor tissue expressed much more genes and had stronger cell-cell interactions than their counterparts in normal tissue. We identified copy number variations (CNVs) in each cancer cell and found correlation between gene copy number and expression level in cancer cells at single cell resolution. Analysis of tumor subclones inferred by CNVs showed accumulation of mutations in each tumor subclone along lineage trajectories. We found differentially expressed genes (DEGs) between tumor subclones had two populations: DEGCNV and DEGreg. DEGCNV, showing high CNV-expression correlation and whose expression differences depend on the differences of CNV level, enriched in housekeeping genes and cell adhesion associated genes. DEGreg, showing low CNV-expression correlation and mainly in low CNV variation regions and regions without CNVs, enriched in cytokine signaling genes. Furthermore, cell-cell communication analyses showed that DEGCNV tends to involve in cell-cell contact while DEGreg tends to involve in secreted signaling, which further support that DEGCNV and DEGreg are two regulatorily and functionally distinct categories.

Preparation of novel polyvinyl alcohol-carbon nanotubes containing imidazolyl ionic liquid/chitosan hydrogel for highly efficient uranium extraction from seawater.

A novel polyvinyl alcohol-carbon nanotube containing an imidazolyl ionic liquid/chitosan composite hydrogel (termed CBCS) was prepared for highly selective uranium adsorption from seawater. The results show that CBCS has good adsorption properties for uranium within the pH range of 5.0-8.0. Kinetics and thermodynamics experiments show that the theoretical maximum adsorption capacity of CBCS to U(VI) is 496.049 mg/g (288 K, pH = 6.0), indicating a spontaneous exothermic reaction. Mechanism analysis shows that the hydroxyl group, amino group, and CN bond on the surface of CBCS directly participate in uranium adsorption and that the dense pores on the surface of CBCS play an important role in uranium adsorption. The competitive adsorption experiment shows that CBCS has excellent uranium adsorption selectivity. In addition, CBCS exhibits good reusability. After five adsorption-desorption cycles, the uranium adsorption rate of CBCS can still reach >98 %. Hence, CBCS has excellent potential for uranium extraction from seawater.

Broadband, Polarization-Sensitive, and Self-Powered High-Performance Photodetection of Hetero-Integrated MoS2 on Lithium Niobate.

High-performance photodetectors hold promising potential in optical communication and imaging systems. However, conventional counterparts are suffering narrow detection range, high power consumption, and poor polarization sensitivity. Characteristics originating from switchable polarization in ferroelectrics can be used to optimize the photo-to-electric procedure and improve the photodetection performance. In this regard, we constructed a configuration by integrating 2-dimensional molybdenum disulfide (MoS2) with ferroelectric lithium niobate (LiNbO3), resulting in the MoS2/LiNbO3 heterostructured photodetector. Benefiting from the pyroelectric effect of LiNbO3, the limitation of bandgap on the detection range can be broken, thus broadening the response band of the detector to 365 to 1,064 nm, as well as enabling the self-powered characteristic. Meanwhile, high carrier mobility and decent light absorbance of MoS2 introduce robust light-matter interactions with the underlying LiNbO3, leading to ultrafast rise/fall times of ≈150 µs/250 µs and switching ratios of up to ≈190. Moreover, the highest responsivity, specific detectivity, and external quantum efficiency achieved were 17.3 A·W-1, 4.3 × 1011 Jones, and 4,645.78%, respectively. Furthermore, because of the anisotropy of the spontaneous-polarized LiNbO3 substrate, the photocurrent of the device achieved a dichroic ratio of 7.42, comparing favorably to most MoS2-based photodetectors. This work demonstrates the integration potential between ferroelectric LiNbO3 and 2-dimensional materials for high-performance photodetection.

Shrimp MANF maintains hemocyte viability via interaction with a tyrosine kinase Abl.

Mesencephalic astrocyte-derived neurotrophic factor (MANF) is a highly conserved cell protective protein. In this study, we explored its functions in shrimp hemocytes. Our results indicated that LvMANF knockdown could cause a decrease in total hemocyte count (THC) and an increase in caspase3/7 activity. To further explore its working mechanism, transcriptomic analyses were performed with wild-type and LvMANF-knockdown hemocytes. Three upregulated genes from transcriptomic data, including FAS-associated factor 2, rho-associated protein kinase 1, and serine/threonine-protein kinase WNK4 were validated with qPCR. Further experiments showed that LvMANF knockdown and tyrosine kinase LvAbl knockdown could decrease tyrosine phosphorylation in shrimp hemocytes. In addition, the interaction between LvMANF and LvAbl was validated with immunoprecipitation. The knockdown of LvMANF would decrease ERK phosphorylation and increase LvAbl expression. Our results suggest intracellular LvMANF may maintain shrimp hemocyte viability by interacting with LvAbl.

Directional Bloch surface wave coupling enabled by magnetic spin-momentum locking of light.

We study the magnetic spin-locking of optical surface waves. Through an angular spectrum approach and numerical simulations, we predict that a spinning magnetic dipole develops a directional coupling of light to transverse electric (TE) polarized Bloch surface waves (BSWs). A high-index nanoparticle as a magnetic dipole and nano-coupler is placed on top of a one-dimensional photonic crystal to couple light into BSWs. Upon circularly polarized illumination, it mimics the spinning magnetic dipole. We find that the helicity of the light impinging on the nano-coupler controls the directionality of emerging BSWs. Furthermore, identical silicon strip waveguides are configured on the two sides of the nano-coupler to confine and guide the BSWs. We achieve a directional nano-routing of BSWs with circularly polarized illumination. Such a directional coupling phenomenon is proved to be solely mediated by the optical magnetic field. This offers opportunities for directional switching and polarization sorting by controlling optical flows in ultra-compact architectures and enables the investigation of the magnetic polarization properties of light.

Nested relation extraction via self-contrastive learning guided by structure and semantic similarity.

The conventional Relation Extraction (RE) task involves identifying whether relations exist between two entities in a given sentence and determining their relation types. However, the complexity of practical application scenarios and the flexibility of natural language demand the ability to extract nested relations, i.e., the recognized relation triples may be components of the higher-level relations. Previous studies have highlighted several challenges that affect the nested RE task, including the lack of abundant labeled data, inappropriate neural networks, and underutilization of the nested relation structures. To address these issues, we formalize the nested RE task and propose a hierarchical neural network to iteratively identify the nested relations between entities and relation triples in a layer by layer manner. Moreover, a novel self-contrastive learning optimization strategy is presented to adapt our method to low-data settings by fully exploiting the constraints due to the nested structure and semantic similarity between paired input sentences. Our method outperformed the state-of-the-art baseline methods in extensive experiments, and ablation experiments verified the effectiveness of the proposed self-contrastive learning optimization strategy.

Deep Learning-Based CT Imaging for the Diagnosis of Liver Tumor.

The objective of this research was to investigate the application value of deep learning-based computed tomography (CT) images in the diagnosis of liver tumors. Fifty-eight patients with liver tumors were selected, and their CT images were segmented using a convolutional neural network (CNN) algorithm. The segmentation results were quantitatively evaluated using the Dice similarity coefficient (DSC), precision, and recall. All the patients were examined and diagnosed by CT enhanced delayed scan technique, and the CT scan results were compared with the pathological findings. The results showed that the DSC, precision, and recall of the CNN algorithm reached 0.987, 0.967, and 0.954, respectively. The images segmented by the CNN were clearer. The diagnostic result of the examination on 56 cases by CT enhanced delay scanning was consistent with that of pathological diagnosis. According to the result of pathological diagnosis, there were 6 cases with hepatic cyst, 9 with hepatic hemangioma, 12 cases with liver metastasis, 10 cases with hepatoblastoma, 3 cases with focal nodular hyperplasia, and 18 cases with primary liver cancer. The result of CT enhanced delay scanning on 58 patients was consistent with that of pathological diagnosis, and the total diagnostic coincidence rate reached 96.55%. In conclusion, the CNN algorithm can perform accurate and efficient segmentation, with high resolution, providing a more scientific basis for the segmentation of liver tumors in CT images. CT enhanced scanning technology has a good effect on the diagnosis and differentiation of liver tumor patients, with high diagnostic coincidence rate. It has important value for the diagnosis of liver tumor and is worthy of clinical application.

Subacute dermal toxicity study of bensulfuron-methyl in Sprague-Dawley rats.

Background: Bensulfuron-methyl has recently attracted attention given its widespread use as an herbicide in crops, especially its transdermal safety. However, no dermal toxicity study of this pesticide to mammals has been reported. The present study aims to investigate subacute dermal toxicity of bensulfuron-methyl following repeated doses exposure.Methods: Forty-eight Sprague-Dawley rats were randomly divided into four groups: a normal control group and bensulfuron-methyl groups of different concentrations (250, 500, 1000 mg/kg.bw/day). The rats were topically applied with the substance dermally for 6 h per day for 28 days. At the end of the experiment, all rats were monitored for any changes in their hematological, biochemical parameters, and pathological and histological sections.Results: There were no statistically significant differences (P ≥ 0.05) in the hematological parameters and biochemical parameters. The pathological histological results of rats in the control and the highest concentration group showed no significant abnormalities. The NOAEL of subacute dermal toxicity study was found to be 1000 mg/kg.bw/day in both female and male rats.Conclusion: The result indicated that bensulfuron-methyl is probably safe for humans as a pesticide.

Computed Tomography Imaging under Artificial Intelligence Reconstruction Algorithm Used in Recovery of Sports Injury of the Knee Anterior Cruciate Ligament.

This study aimed to analyze the influence of artificial intelligence (AI) reconstruction algorithm on computed tomography (CT) images and the application of CT image analysis in the recovery of knee anterior cruciate ligament (ACL) sports injuries. A total of 90 patients with knee trauma were selected for enhanced CT scanning and randomly divided into three groups. Group A used the filtered back projection (FBP) reconstruction algorithm, and the tube voltage was set to 120 kV during CT scanning. Group B used the iDose4 reconstruction algorithm, and the tube voltage was set to 120 kV during CT scanning. In group C, the iDose4 reconstruction algorithm was used, and the tube voltage was set to 100 kV during CT scanning. The noise, signal-to-noise ratio (SNR), carrier-to-noise ratio (CNR), CT dose index volume (CTDI), dose length product (DLP), and effective radiation dose (ED) of the three groups of CT images were compared. The results showed that the noise of groups B and C was smaller than that of group A (P < 0.05), and the SNR and CNR of groups B and C were higher than those of group A. The images of patients in group A with the FBP reconstruction algorithm were noisy, and the boundaries were not clear. The noise of the images obtained by the iDose4 reconstruction algorithm in groups B and C was improved, and the image resolution was also higher. The agreement between arthroscopy and CT scan results was 96%. Therefore, the iterative reconstruction algorithm of iDose4 can improve the image quality. It was of important value in the diagnosis of knee ACL sports injury.

Comparative Pharmacokinetic Study of Three Major Bioactive Components in the Normal and Different Courses of Liver-Stagnation and Spleen-Deficiency Syndrome Depressive Rats after Intragastric Administration of Zhi-Zi-Hou-Po Decoction.

To reveal the pharmacokinetic process of narirutin, naringin, and honokiol in normal and different courses of liver-stagnation and spleen-deficiency syndrome depressive rats after intragastric administration of Zhi-Zi-Hou-Po decoction (ZZHPD), a rapid ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) method was proposed in this study. Chronic unpredictable mild stress (CUMS) rat was employed as the depression model. Acetonitrile solution containing 0.1% acetic acid and methyl alcohol-water (50 : 50, v/v) was chosen as the protein precipitant and redissolve solution severally; a Shim-pack GISS C18 column coupled with 0.1% aqueous acetic acid-acetonitrile in gradient elution was employed to separate the mixed constituents in plasma. The WinNonLin software (version 6.1) was chosen as the analytical tool for the pharmacokinetic parameters. The results indicated that compared with rats in the control group, the sucrose preference, scores in the open-field test, and the concentration of 5-HT in plasma of rats in CUMS and CUMS + ZZHPD treatment groups were lower, while the immobility time in the forced swimming test of rats in these two groups was longer, which implied that the depression model was successful. These behavioral and biochemical indexes of rats above in the CUMS + ZZHPD treatment group were improved after oral administration of ZZHPD, which indicated that the antidepressant effect of ZZHPD was definite. The UHPLC-MS/MS method was stabilized, sensitive, and exclusive, and the extraction recovery and matrix effect of three analytes were all above 89%. The T max, AUC, and Cmax of three ingredients in CUMS-induced depression rats were significantly larger than control rats, while these pharmacokinetic parameters in CUMS + ZZHPD treatment rats were decreased significantly compared with CUMS-induced depression rats, which may relate with the changes in physiological function of the gastrointestinal tract and liver in CUMS-induced liver-stagnation and spleen-deficiency syndrome depressive rats. This study provided important information for the clinical rational use of ZZHPD in antidepressant treatment.

Electro-optic high-speed optical beam shifting based on a lithium niobate tapered waveguide.

We propose an electro-optic on-chip beam shifting device based on gradient microstructured electrodes and an optical tapered waveguide fabricated using lithium niobate (LN). The distribution of refractive index variations of the optical waveguide can be electro-optically defined and tailored by the designed gradient microstructured electrodes, which directs the beam propagation and shifting. The length of the beam shifting device is 18 mm and the width of the waveguide is gradually increased from 8 µm to 80 µm. The functionality of the beam shifting device is experimentally demonstrated, and it is observed that it has an electro-optic tunability of 0.41 µm/V, and a high-speed response time of 19 ns (λ=1310 nm). This study can provide potential applications in optical switching and modulation, beam scanning and ranging, optical spatial communications, etc.

[Open reduction and internal fixation with plate via posteromedial approach of retaining pes anserinus tendon in the treatment of tibial plateau fracture].

OBJECTIVE: To compare the effects of cutting and retaining the pes anserinus tendon on effectiveness following tibial plateau fracture. METHODS: A clinical data of 40 patients with tibial plateau fracture treated with open reduction and internal fixation with plate via posteromedial approach between January 2015 and January 2020 was retrospectively analyzed, including 18 patients retained the pes anserinus tendon (study group) and 22 patients cut the pes anserinus tendon (control group) during operation. There was no significant difference in gender, age, side of affected knee, cause of injury, Schatzker classification, time from injury to operation, and associated ligament injury between the two groups ( P>0.05). The operation time, intraoperative blood loss, hospital stay, anatomic reduction rate, incidence of complications, fracture healing time, knee flexion and extension range of motion at 2 weeks and 12 months, and knee extension range of motion at 3 months after operation were recorded and compared between the two groups. The visual analogue scale (VAS) score was used to evaluate the early postoperative pain improvement at 1, 3, and 14 days after operation and hospital for special surgery (HSS) score was used to evaluate the improvement of knee function at 3, 6, and 12 months after operation. RESULTS: The patients in both groups were followed up 12-15 months with an average of 12.8 months. There was no significant difference in operation time, intraoperative blood loss, and fracture healing time between the two groups ( P>0.05). The hospital stay in the control group was significantly longer than that in the study group ( t=8.339, P=0.000). There was no significant difference in the anatomic reduction rate (90.9% vs. 83.3%) between the control group and the study group ( χ 2=0.058, P=0.810). There were 1 case of proximal tibial osteomyelitis, 3 cases of skin necrosis, 3 cases of traumatic arthritis, and 2 cases of lower deep venous thrombosis after operation in the control group, and 1 case of metaphyseal nonunion, 2 cases of traumatic arthritis, and 1 case of lower deep venous thrombosis in the study group, showing no significant difference in the incidence of complications (40.9% vs. 22.2%) between the two groups ( χ 2=1.576, P=0.209). In the study group, knee flexion and extension range of motion at 2 weeks and 12 months and knee extension range of motion at 3 months after operation were significantly better than those of the control group ( P<0.05). VAS scores and HSS scores in both groups improved with time after operation ( P<0.05), in addition, the HSS score and VAS score of the study group were significantly better than those of the control group ( P<0.05). CONCLUSION: Compared with traditional pes anserinus tendon cutting group, pes anserinus tendon retaining group can significantly reduce postoperative short-term pain, improve postoperative knee range of motion and knee function within 1 year after operation.

Shrimp Plasma MANF Works as an Invertebrate Anti-Inflammatory Factor via a Conserved Receptor Tyrosine Phosphatase.

For a long time, how anti-inflammatory factors evolved was largely unknown. In this study, we chose a marine invertebrate, Litopenaeus vannamei, as a model and identified that shrimp mesencephalic astrocyte-derived neurotrophic factor (MANF) was an LPS-induced plasma protein, which exerted its anti-inflammatory roles on shrimp hemocytes by suppressing ERK phosphorylation and Dorsal expression. In addition, we demonstrated that shrimp MANF could be associated with a receptor protein tyrosine phosphatase (RPTP) to mediate negative regulation of ERK activation and Dorsal expression. More interestingly, shrimp RPTP-S overexpression in 293T cells could switch shrimp and human MANF-mediated ERK pathway activation to inhibition. In general, our results indicate that this conserved RPTP is the key component for extracellular MANF-mediated ERK pathway inhibition, which gives a possible explanation about why this neurotropic factor could both protect neuron cells from apoptosis and inhibit immune cell M1 activation in various species.

Identifying the effectual-combination ingredients of Zhi-zi-Hou-po decoction based on metabolic difference-oriented network regulation strategy.

Zhi-zi-Hou-po decoction (ZZHPD) has been used to treat depression in the clinic for thousand years in China. However, the pharmacodynamic substance of ZZHPD is still not totally clear due to its complex components. The objective of this study was to identify the effectual combination ingredients (ECIs) of ZZHPD, which could represent the antidepressant effect of the original formula. Firstly, differential plasma absorbed components with different variable importance in projection (VIP) value in chronic unpredictable mild stress (CUMS)-induced depression and control rat were revealed by untargeted metabolomics-driven strategy based on HPLC-ESI-TOF/MS, XCMS online and SIMCA-p software. Secondly, network topology scores (NTS) of plasma absorbed components were exposed by protein-protein interaction (PPI) network analysis based on components-related genes and depression-related genes, which were performed by network pharmacology tools. Finally, the ECIs were screened by considered of VIP value and NTS. Then the bioactivity was evaluated by cell viability and expression of glial fibrillary acid protein (GFAP), tumor necrosis factor α (TNFα) and interleukin 1ß (IL-1ß) of a lipopolysaccharide-induced astrocyte depression model. An effective combination composed with 12 components was filtrated as ECIs of ZZHPD, exposed to which the cell viability effect, the expression of GFAP and IL-1ß in astrocytes were essentially equivalent with original ZZHPD (p > 0.05), and that both characteristic constituents and trace compounds of ZZHPD might exert synergistic actions through multi-targets. The result of this study provided useful information for the clinical application and modern development of ZZHPD, and the proposed strategy could be regard as an alternative solution for effective combination screening of herbal medicines.

CRISPR-mediated live imaging of genome editing and transcription.

We report a robust, versatile approach called CRISPR live-cell fluorescent in situ hybridization (LiveFISH) using fluorescent oligonucleotides for genome tracking in a broad range of cell types, including primary cells. An intrinsic stability switch of CRISPR guide RNAs enables LiveFISH to accurately detect chromosomal disorders such as Patau syndrome in prenatal amniotic fluid cells and track multiple loci in human T lymphocytes. In addition, LiveFISH tracks the real-time movement of DNA double-strand breaks induced by CRISPR-Cas9-mediated editing and consequent chromosome translocations. Finally, by combining Cas9 and Cas13 systems, LiveFISH allows for simultaneous visualization of genomic DNA and RNA transcripts in living cells. The LiveFISH approach enables real-time live imaging of DNA and RNA during genome editing, transcription, and rearrangements in single cells.

Quantitative serum proteomics analyses reveal shrimp responses against WSSV infection.

For the past decades, quantitative serum proteomics in mammals have been widely applied in biomarker screening. Various serum protein removal methods have been developed to effectively sequester serum abundant proteins. However, few methods have been found for the removal of arthropod serum abundant proteins. Here, gel filtration chromatography and ultracentrifugation methods were applied to remove hemocyanin from Litopeaneus vannamei serum. When shrimps were challenged with white spot syndrome virus (WSSV), a total of 486 serum proteins were identified using mass spectrometry, and 18 upregulated WSSV responsive proteins were identified with isobaric tags for relative and absolute quantification (iTRAQ). These results provide an effective method to remove hemocyanin from shrimp serum. With this method some previously unidentified WSSV responsive serum proteins were revealed, which would give us a better insight into the response of crustaceans to WSSV infection.

Metabolic profile analysis of Zhi-zi-chi decoction in feces of normal and chronic unpredictable mild stress-induced depression rats based on UHPLC-ESI-Q-TOF-MS/MS and multiple analytical strategies.

Zhi-zi-chi decoction (ZZCD) has been verified by clinical application that it has definite curative effects and low side effects on depression. Because it is administered orally, the metabolites of ZZCD in the intestinal tract may influence the curative effects significantly. In this study, UHPLC-ESI-Q-TOF-MS/MS was used in combination with untargeted metabolomics-driven strategy, series product ion filtering and diagnostic fragment ion strategy for acquiring the comprehensive metabolic profile of ZZCD in feces of normal and chronic unpredictable mild stress (CUMS)-induced depression rats after oral administration, while the rat depression model was evaluated by behavior tests and plasma biochemical indices. Finally, a total of 56 compounds, including 35 prototype compounds and 21 metabolites, were identified or tentatively characterized in fecal samples. Among these, ten compounds were sieved as potential chemical markers that would reflect the antidepressant effect of ZZCD, which may offer important information for quality assessment, pharmacokinetic study and clinical security. In conclusion, the metabolic profile of ZZCD in normal and CUMS-induced depression rats would be helpful for the further study of anti-depression material basis and mechanism.

Characterization of global metabolic profile of Zhi-Zi-Hou-Po decoction in rat bile, urine and feces after oral administration based on a strategy combining LC-MS and chemometrics.

Identification of metabolic profile of traditional Chinese medicine in vivo is always a challenge task. Usually, screening out and identifying the exogenous compounds manually from total ion chromatograms (TICs) of biologic samples is time-consuming and strenuous. In this study, a systematic identification strategy based on LC-MS was adopted to clarify the metabolic profiling of Zhi-Zi-Hou-Po decoction (ZZHPD) in rat. Bile, urine and feces samples of rat were obtained after oral administration and then analyzed by LC-MS after proper preparation. The xenobiotics were screened out from TICs globally and rapidly by untargeted metabolomics-driven strategy (UMDS) based on the combined of XCMS online (a web-based platform to process LC-MS data), MetAlign (a software to process LC-MS data) and SIMCA-P (a software for data analysis). Most of the xenobiotics were identified by means of series product ions filtering (sPIF), which was based on the database-hit of ZZHPD (including prototype and metabolites). Then the unmatched constituents were identified tentatively and their source and metabolic pathway were clarified by using diagnostic fragment ions strategy (DFIS). As a result, a total of 83 compounds including 44 prototype compounds and 39 metabolites were rapidly identified or tentatively characterized from the biologic samples, and among them, four of which were found for the first time. Further research on the correlations of these prototype compounds and metabolites revealed that glucuronidation is the main metabolic pathways of ZZHPD in rat bile and urine, while prototype compounds were the abundant ingredients detected in rat feces.

Identification of absorbed components and metabolites of Zhi-Zi-Hou-Po decoction in rat plasma after oral administration by an untargeted metabolomics-driven strategy based on LC-MS.

In this study, a novel untargeted metabolomics-driven strategy based on LC-MS was used to rapidly screen and identify the absorbed components and metabolites of Zhi-Zi-Hou-Po decoction (ZZHPD) in rat plasma. The plasma sample was obtained from orbital venous of rats after oral administration and pretreated by protein precipitation before analysis. All sample data from total ion chromatograms (TICs) of LC-TOF/MS were aligned and peak picked by XCMS and MetAlign combined to extract three-dimensional datasets (peak code, t R -m/z pairs and ion intensity). Xenobiotics in rat plasma were differentiated from endogenous components by multivariate statistical analysis and then divided into prototype compounds and metabolites by comparing t R -m/z with the chemical compounds of ZZHPD. Combined with fragment ions and structure information of LC-TSQ/MS, a total of 61 compounds, including 35 prototype compounds and 26 metabolites, were rapidly identified or tentatively characterized in rat plasma. Results indicated that iridoid glycosides, monoterpenoids, flavonoids, and lignans were the main absorbed chemical components of ZZHPD. Glucuronidation and sulfation were the main metabolic pathways of ZZHPD compounds in vivo. In addition, there were ring-opening reactions and reduction reactions for iridoid glycosides, hydrolysis for flavonoids, as well as hydroxylation and stereoscopic conversion reactions for lignans. This study offers a systematically applicable approach for rapid screening and identification of xenobiotics and metabolites derived from multi-herb prescription in vivo, and provides useful information for ascertaining bioactive ingredients and action mechanisms of ZZHPD. Graphical Abstract Diagram of untargeted metabolomics-driven strategy for ZZHPD in rat plasma.