Rab27B enhances drug resistance in hepatocellular carcinoma by promoting exosome-mediated drug efflux.

Liver cancer is a major threat to human life and health, and chemotherapy has been the standard non-surgical treatment for liver cancer. However, the emergence of drug resistance of liver cancer cells has hindered the therapeutic effect of chemical drugs. The discovery of exosomes has provided new insights into the mechanisms underlying tumour cell resistance. In this study, we aimed to determine the proteins associated with drug resistance in tumour cells and to elucidate the underlying mechanisms. We found that Rab27B expression in drug (5-fluorouracil, 5Fu)-resistant Bel7402 (Bel/5Fu) cells increased significantly compared with that in drug-sensitive Bel7402 cells. In addition, Bel/5Fu cells secreted more exosomes under 5Fu stimulation. The number of exosomes secreted by Bel/5Fu cells significantly reduced after knocking down Rab27B, and the cellular concentration of 5Fu increased, enhancing its therapeutic effect. We also found that the administration of classical drug efflux pump (P-glycoprotein, P-gp) inhibitors together with knockdown of Rab27B further improved the therapeutic effects of chemotherapy drugs. In conclusion, our findings suggest that Rab27B could be a new therapeutic target in liver cancer.

The critical role of exosomes in tumor biology.

Exosomes are a type of extracellular vesicles (diameter, 30-160 nm), which contain multiple proteins, nucleic acids, lipid molecules, and other substances. Most types of cells can secrete exosomes, although the biogenesis, composition, and function is specific to different cell types. Recently, many studies have demonstrated that exosomes play a critical role in tumor development. In this review, we briefly summarize the biogenesis, composition, and function of exosomes. We also discuss the recent advances in the critical role of exosomes in tumor biology with a special focus on their application in tumor diagnosis and treatment.

Molecularly Imprinted Poly(thionine)-Based Electrochemical Sensing Platform for Fast and Selective Ultratrace Determination of Patulin.

An innovative approach based on a surface functional monomer-directing strategy for the construction of a sensitive and selective molecularly imprinted electrochemical sensor for patulin recognition is described. A patulin imprinted platinum nanoparticle (PtNP)-coated poly(thionine) film was grown on a preformed thionine tailed surface of PtNP-nitrogen-doped graphene (NGE) by electropolymerization, which provided high capacity and fast kinetics to uptake patulin molecules. Thionine acted not only as a functional monomer for molecularly imprinted polymer (MIP), but also as a signal indicator. Enhanced sensitivity was obtained by combining the excellent electric conductivity of PtNPs, NGE, and thionine with multisignal amplification. The designed sensor displayed excellent performance for patulin detection over the range of 0.002-2 ng mL-1 (R2 = 0.995) with a detection limit of 0.001 ng mL-1 for patulin. In addition, the resulting sensor showed good stability and high repeatability and selectivity. Furthermore, the feasibility of its applications has also been demonstrated in the analysis of real samples, providing novel tactics for the rational design of MIP-based electrochemical sensors to detect a growing number of deleterious substances.

The Nogo-B receptor promotes human hepatocellular carcinoma cell growth via the Akt signal pathway.

Nogo-B receptor (NgBR) is a type I receptor with a single transmembrane domain and specifically binds to ligand Nogo-B. A previous study demonstrated that NgBR was highly expressed in human breast invasive ductal carcinoma and promoted epithelial-mesenchymal transition in breast tumor cells. Our recent work found that NgBR expression was associated with a poor prognosis in human patients with hepatocellular carcinoma (HCC). Here, we elucidate that the increased expression of NgBR contributes toward the increased cell growth of human HCC cells both in vitro and in vivo. Cell viability and clonogenic survival analysis results demonstrated that knockdown of NgBR inhibits the cell growth in human HCC cells, which correlates with a reduction in the phosphorylation of Akt levels. Furthermore, overexpression of NgBR by the cotransfected pIRES-NgBR plasmid together with NgBR siRNA in human HCC cells can rescue impaired phosphorylation of Akt levels in NgBR knockdown human HCC cells. In addition, cell viability analyses showed that NgBR overexpression can rescue the cell growth inhibition presented in human HCC NgBR knockdown cells. Taken together, our results suggest that NgBR potentially acts as an oncogene in HCC by increasing Akt activity. Thus, NgBR may represent a new potential diagnostic and therapeutic target for the treatment of HCC.

Multipotent mesenchymal stromal cells play critical roles in hepatocellular carcinoma initiation, progression and therapy.

Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer, with high morbidity, relapse and mortality rates. Multipotent mesenchymal stromal cells (MSCs) can be recruited to and become integral components of the HCC microenvironment and can influence tumor progression. This review discusses MSC migration to liver fibrosis and the HCC microenvironment, MSC involvement in HCC initiation and progression and the widespread application of MSCs in HCC-targeted therapy, thus clarifying the critical roles of MSCs in HCC.

Lipid-related FABP5 activation of tumor-associated monocytes fosters immune privilege via PD-L1 expression on Treg cells in hepatocellular carcinoma.

Monocytes/macrophages, a plastic and heterogeneous cell population of the tumor microenvironment (TME), can constitute a major component of most solid tumors. Under the pressure of rapid proliferation of the tumor, monocytes/macrophages can be educated and foster immune tolerance via metabolic reprogramming. Our studies have shown that the activation of FABP5, a lipid-binding protein, decreases the rate of ß-oxidation causing the accumulation of lipid droplets in monocytes. We found that hepatocellular carcinoma cells (HCC) increased IL-10 secretion by monocytes, which depended on the expression of FABP5 and suppressing of the PPARα pathway. Moreover, the elevated level of IL-10 promotes PD-L1 expression on Treg cells via the JNK-STAT3 pathway activation. We also observed that elevation of FABP5 in monocytes was negatively related to HCC patients' overall survival time. Thus, FABP5 promotes monocyte/macrophage lipid accumulation, fosters immune tolerance formation, and might represent itself as a therapeutic target in both tumor-associated monocytes (TAMs) and cancer cells.

Exposure Assessment of Multiple Mycotoxins and Cumulative Health Risk Assessment: A Biomonitoring-Based Study in the Yangtze River Delta, China.

The extensive exposure to multiple mycotoxins has been demonstrated in many countries; however, realistic assessments of the risks related to cumulative exposure are limited. This biomonitoring study was conducted to investigate exposure to 23 mycotoxins/metabolites and their determinants in 227 adults (aged 20-88 years) in the Yangtze River Delta, China. Eight mycotoxins were detected in 110 urine samples, and multiple mycotoxins co-occurred in 51/227 (22.47%) of urine samples, with deoxynivalenol (DON), fumonisin B1 (FB1), and zearalenone (ZEN) being the most frequently occurring. For single mycotoxin risk assessment, FB1, ZEN, aflatoxin B1 (AFB1), and ochratoxin A (OTA) all showed potential adverse effects. However, for the 12 samples containing DON and ZEN, in which none had a hazard risk, the combination of both mycotoxins in two samples was considered to pose potential endocrine disrupting risks to humans by hazard index (HI) method. The combined margin of exposure (MOET) for AFB1 and FB1 could constitute a potential health concern, and AFB1 was the main contributor. Our approach provides a blueprint for evaluating the cumulative risks related to different types of mycotoxins and opens a new horizon for the accurate interpretation of epidemiological health outcomes related to multi-mycotoxin exposure.

Dual-target electrochemical aptasensor based on co-reduced molybdenum disulfide and Au NPs (rMoS2-Au) for multiplex detection of mycotoxins.

Multiple mycotoxin contamination has posed health risks in the area of food safety. In this study, co-reduced molybdenum disulfide and gold nanoparticles (rMoS2-Au) were designed and used for the first time as an efficient platform endowing electrochemical electrodes with superior electron transfer rates, large surface areas and strong abilities to firmly couple with large amounts of different aptamers. After further modification with thionine (Thi) and 6-(Ferrocenyl) hexanethiol (FC6S), a platform enabling sensitive, selective and simultaneous determination of two important mycotoxins, zearalenone (ZEN) and fumonisin B1 (FB1), was achieved. The established aptasensor showed excellent linear relationships (R2 > 0.99) when ZEN and FB1 concentrations were in the range of 1 × 10-3-10 ng mL-1 and 1 × 10-3-1 × 102 ng mL-1, respectively. High sensitivity of ZEN and FB1 with a limit of detection as low as 5 × 10-4 ng mL-1 was obtained with excellent selectivity and stability. The effectiveness of the aptasensor was verified in real maize samples, and satisfactory recoveries were attained. The established platform could be easily expanded to other aptamer-based multiplex screening protocols in biochemical research and clinical diagnosis.

Exosome-derived ENO1 regulates integrin α6ß4 expression and promotes hepatocellular carcinoma growth and metastasis.

Alpha-enolase (ENO1) has been found to be dysregulated in several human malignancies, including hepatocellular carcinoma (HCC). Although the role of ENO1 as a glycolytic enzyme in HCC cells has been well characterized, little is known about the other roles of ENO1, especially exosome-derived ENO1, in regulating HCC progression. Here, we demonstrated that ENO1 is frequently upregulated in HCC cells or tissues, with even higher expression in highly metastatic HCC cells or metastatic tissues as well as in exosomes derived from highly metastatic sources. Moreover, ENO1 expression is associated with the tumor-node-metastasis (TNM) stage, differentiation grade and poor prognosis in HCC patients. Surprisingly, ENO1 can be transferred between HCC cells via exosome-mediated crosstalk, exhibiting an effect similar to that of ENO1 overexpression in HCC cells, which promoted the growth and metastasis of HCC cells with low ENO1 expression by upregulating integrin α6ß4 expression and activating the FAK/Src-p38MAPK pathway. In summary, our data suggest that exosome-derived ENO1 is essential to promoting HCC growth, metastasis, and further patient deterioration. The findings from this study implicate a novel biomarker for the clinical evaluation of HCC progression, especially the prediction of HCC metastatic risk.

Reduced graphene oxide-zinc oxide nanocomposite as dispersive solid-phase extraction sorbent for simultaneous enrichment and purification of multiple mycotoxins in Coptidis rhizoma (Huanglian) and analysis by liquid chromatography tandem mass spectrometry.

In the current study, a robust dispersive solid-phase extraction (dSPE) strategy using reduced graphene oxide-zinc oxide (rGO-ZnO) nanocomposite as the sorbent was proposed for separation, purification and enrichment of 12 mycotoxins in Coptidis rhizoma (Huanglian). The targeted mycotoxins included aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, aflatoxin M1, alternariol-methylether, mycophenolic acid, ochratoxin A, penitrem A, nivalenol, zearalenone and zearalanone. The rGO-ZnO nanocomposite was successfully synthesized through hydrothermal process by a modified Hummers method, and further characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), FTIR spectroscopy, ultraviolet-visible spectroscopy and X-ray diffraction (XRD). Several key parameters affecting the performance of the dSPE approach were extensively investigated, and after optimization, acetonitrile/water/formic acid (80/19/1, v/v/v) as the extraction solution, 2% acetonitrile as the adsorption solution, 15 mg rGO-ZnO as the sorbent, n-hexane as the washing solution, and methanol/formic acid (99/1, v/v) as the desorption solution presented an excellent purification and enrichment efficiency. Under the optimal dSPE procedure followed by analysis with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), adequate linearity (R2 ≥ 0.991), high sensitivity (limit of quantification in the range of 0.09-0.41 µg kg-1), acceptable recovery (70.3-105.7%) and satisfactory precision (RSD 1.4-15.0%) were obtained. The analysis of 12 selected mycotoxins was also carried out in real Coptidis rhizoma (Huanglian) samples for applicability evaluation of the established method.

Thin-layer MoS2 and thionin composite-based electrochemical sensing platform for rapid and sensitive detection of zearalenone in human biofluids.

In this study, a facile electrochemical sensing platform based on thin-layer MoS2 and thionin (MoS2-Thi) composites was constructed for the sensitive and rapid detection of zearalenone (ZEA) in human biofluids. MoS2-Thi composites with a thin-layer MoS2 structure and large specific surface area were synthesized via a simple ultrasound exfoliation method. To further enhance the performance of the platform but without increasing the complexity of the fabrication process, ZEA antibodies were modified with Pt nanoparticles (Pt NPs) using an environmentally friendly method, where they had a much wider linear range and four times higher sensitivity compared with the original ZEA monoclonal antibody (ZEA-MAb). Under the optimal conditions, the electrochemical responses of the MoS2-Thi composite-based immunosensor were linear (R2 = 0.9915) when the ZEA concentrations ranged from 0.01 to 50 ng mL-1. The proposed immunosensor could detect ZEA at concentrations as low as 0.005 ng mL-1 with excellent selectivity. The immunosensor exhibited acceptable stability with high reproducibility and accuracy, and it was applied successfully for determining ZEA in human plasma and urine samples. Our findings indicated that the proposed MoS2-Thi composite-based electrochemical sensing platform provides a new approach for the rapid and sensitive bioanalysis of ZEA, and it may also be extended to other mycotoxins for multiplex detection.

Label-Free Fluorescent Aptasensor for Ochratoxin-A Detection Based on CdTe Quantum Dots and (N-Methyl-4-pyridyl) Porphyrin.

With the widespread contamination of ochratoxin A (OTA), it is of significant importance for detecting OTA in foods and traditional Chinese medicine (TCM). In this study, a novel label-free fluorescent aptasensor utilizing the interaction between OTA-triggered antiparallel G-quadruplex and (N-methyl-4-pyridy) porphyrin (TMPyP) for the rapid and sensitive determination of OTA was established. The fluorescence of CdTe quantum dots (QDs) could be quenched by TMPyP. In the presence of analyte (OTA), the aptamer could recognize OTA and transform from a random coil to the antiparallel G-quadruplex. The interaction between G-quadruplex and TMPyP could release CdTe QDs from TMPyP, and thus recover the fluorescence of CdTe QDs. Under optimized conditions, the detection limit of the designed aptasensor was 0.16 ng mL-1, with a linear range of 0.2 to 20 ng mL-1. Furthermore, this aptasensor showed high selectivity toward OTA against other structural analogs and other mycotoxins, and was successfully applied in Astragalus membranaceus samples. The presented aptasensor for OTA detection could be a promising tool for the field monitoring of food and TCM.

Heterogeneity of cancer-associated fibroblasts and roles in the progression, prognosis, and therapy of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a lethal disease, and recurrence and metastasis are the major causes of death in HCC patients. Cancer-associated fibroblasts (CAFs), a major stromal cell type in the HCC microenvironment, promote HCC progression, and have gradually become a hot research topic in HCC-targeted therapy. This review comprehensively describes and discusses the heterogeneous tissue distribution, cellular origin, phenotype, and biological functions of HCC-associated fibroblasts. Furthermore, the possible use of CAFs for predicting HCC prognosis and in targeted therapeutic strategies is discussed, highlighting the critical roles of CAFs in HCC progression, diagnosis, and therapy.

Determination of multiple mycotoxins in paired plasma and urine samples to assess human exposure in Nanjing, China.

This study was conducted to investigate mycotoxin exposure in 260 rural residents (age 18-66 years) in Nanjing, China. Paired plasma and first morning urine samples were analyzed for 26 mycotoxin biomarkers, including 12 parent mycotoxins and 14 mycotoxin metabolites, by an ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method. Mycotoxins and their metabolites were detected in 95/260 (36.5%) plasma samples and 144/260 (55.4%) urine samples. The most prevalent mycotoxin in plasma was ochratoxin A (OTA), with the incidence of 27.7% (range 0.312-9.18 µg/L), while aflatoxin B1-lysine (AFB1-lysine) (incidence 19.6%, range 10.5-74.5 pg/mg albumin), fumonisin B1 (FB1) (incidence 2.7%, range 0.305-0.993 µg/L), deoxynivalenol (DON) (incidence 2.3%, range 1.39-5.53 µg/L), zearalenone (ZEN) (incidence 6.5%, range 0.063-0.418 µg/L) and zearalanone (ZAN) (incidence 1.2%, range 0.164-0.346 µg/L) were also detected in plasma samples. Deoxynivalenol-15-glucuronide (DON-15-GlcA) was the most frequently detected urinary mycotoxin, with the incidence of 43.8% (range 0.828-37.7 µg/L). DON (incidence 10.0%, range 1.39-14.7 µg/L), DON-3-glucuronide (DON-3-GlcA) (incidence 15.8%, range 0.583-5.84 µg/L), aflatoxin M1 (AFM1) (incidence 10.4%, range 0.125-0.464 µg/L), ZAN (incidence 7.7%, range 0.106-1.82 µg/L), ZEN (incidence 6.9%, range 0.056-0.311 µg/L), FB1 (incidence 3.1%, range 0.230-1.33 µg/L), T-2 toxin (incidence 2.3%, range 0.248-3.61 µg/L) and OTA (incidence 1.2%, range 0.153-0.557 µg/L) were also found in urine samples. Based on the plasma or urinary levels, the daily intakes of AFB1, FB1, ZEN, DON and OTA were estimated. The results showed that the investigated rural dwellers were exposed to multiple mycotoxins, especially to carcinogenic mycotoxin AFB1 with a mean daily intake of 0.41 µg/kg·bw/day, thereby underlining a potential public health concern. To the best of our knowledge, this is the first study to evaluate human exposure to mycotoxins with direct measurements of multiple mycotoxins in paired plasma and urine samples for over 200 subjects of a single population.

CBX2 Regulates Proliferation and Apoptosis via the Phosphorylation of YAP in Hepatocellular Carcinoma.

Chromobox 2 (CBX2), a chromobox family protein, is a crucial component of the polycomb group complex: polycomb repressive complex 1 (PRC1). Research on CBX2 as an oncogene has been published in recent years. However, the connection between CBX2 and hepatocellular carcinoma (HCC) has not been studied. In this article, based on the results of immunohistochemical (IHC) staining of HCC and adjacent liver tissue microarrays, we found that high CBX2 expression is associated with poor prognosis in HCC patients. The results of a CCK8 assay, a clonogenic survival assay and a nude mouse tumorigenicity assay showed that knockdown of CBX2 inhibited the proliferation of HCC cells. According to the results of Annexin V-FITC/propidium iodide (PI) staining-based fluorescence activated cell sorting (FACS) analysis, knockdown of CBX2 increased HCC cell apoptosis. Furthermore, the RNA-seq results revealed that knockdown of CBX2 inhibited the expression of WTIP, which is an inhibitor of the Hippo pathway. We used western blotting to validate the mechanism and discovered that knockdown of CBX2 increased the phosphorylation of YAP, which explains why knockdown of CBX2 inhibits proliferation and increases apoptosis in HCC cells. In conclusion, CBX2 could be a potential target for HCC anticancer treatment.

Reduced graphene oxide and gold nanoparticle composite-based solid-phase extraction coupled with ultra-high-performance liquid chromatography-tandem mass spectrometry for the determination of 9 mycotoxins in milk.

A reliable solid-phase extraction (SPE) procedure using reduced graphene oxide and gold nanoparticle (rGO/Au) composite as sorbent was proposed for purification and enrichment of aflatoxin B1 (AFB1), aflatoxin M1 (AFM1), ochratoxin A (OTA), zearalenone (ZEA), α-zearalenol (α-ZOL), ß-zearalenol (ß-ZOL), zearalanone (ZAN), α-zearalanol (α-ZAL) and ß-zearalanol (ß-ZAL) in milk. Main parameters affecting the performance of SPE procedure were thoroughly investigated. The optimized conditions included 2% acetonitrile in water as the loading solution, 5% methanol in water as the washing solution, and methanol/acetonitrile/formic acid (50/49/1, v/v/v) as the elution solvent. Satisfactory linearity (R2 ≥ 0.992), high sensitivity (limit of quantification in the range of 0.02-0.18 ng mL-1), adequate recovery (70.2-111.2%), and acceptable precision (RSD, 2.0-14.9%) were obtained when the optimal sample pretreatment protocol was combined with ultra-high-performance liquid chromatography-tandem mass spectrometry detection. The applicability of the validated method was further verified in real milk samples.

Fluorescence imaging of hepatocellular carcinoma with a specific probe of COX-2.

Hepatocellular carcinoma (HCC) is the major subtype of primary liver cancer. Although the standard treatment method based on surgery has generally extended life, it still causes the second and sixth most prevalent cancer-related death in men and women, respectively. The recurrence of cancer caused by unclear resection margins and any remaining undiscovered metastatic nodules should take a large proportion of responsibility for the poor prognosis after resective surgery. Therefore, a practical and effective method that can be used during hepatectomy to specifically identify HCC is a potentially significant area deserving attention. Tests involving fluorescence have been used in many biological systems. In this study, we use a probe that can combine with cyclooxygenase-2 (COX-2) and subsequently emit fluorescence to identify HCC cells and heteroplastic tumors in a mouse model. The results show that this specific probe can clearly differentiate HCC, with differences that could be observed with the naked eye in human samples. The biotechnology of knocking down COX-2 and its inhibitor were used on human HCC cell line SMMC7721, and the outcomes confirmed the above results. The toxic effect also showed that the probe had no harmful effect on normal liver cells. Taken together, our study demonstrates that a COX-2-specific fluorescence probe may be a new and effective method to identify HCC, especially during surgery.

Iron (II, III) oxide/multi-walled carbon nanotube composite as solid-phase extraction sorbent followed by ultra-high performance liquid chromatography tandem mass spectrometry for simultaneous determination of zearalenone and type A trichothecenes in Salviae miltiorrhizae Radix et Rhizoma (Danshen).

For the first time, a reliable solid-phase extraction (SPE) procedure using iron (II, III) oxide (Fe3O4)/multi-walled carbon nanotube (MWCNT) composite as sorbents was proposed for purification and enrichment of zearalenone (ZEA) and four type A trichothecenes (T-2 toxin (T-2), HT-2 toxin (HT-2), neosolaniol (NEO) and diacetoxyscirpenol (DAS)) in Salviae miltiorrhiza Radix et Rhizoma (Danshen). The Fe3O4/MWCNT composite was synthesized by assembling Fe3O4 with MWCNT by sonication through an "aggregation wrap" mechanism and several key parameters affecting the performance of SPE procedure such as the composition of sample loading solutions, washing and elution solvents were thoroughly investigated. After optimization, 2% acetonitrile aqueous solution as the loading solution, 5% methanol aqueous solution as the washing solution and acetone containing 0.5% formic acid as the elution solvent presented an excellent purification efficiency for the five targets in Danshen. Under the optimal sample pretreatment conditions followed by analysis with ultra-high performance liquid chromatography-tandem mass spectrometry, satisfactory linearity (R2≥0.991), high sensitivity (limit of quantification in the range of 1.20-4.80µgkg-1), good recovery (73.7-91.9%) and acceptable precision (RSD, 2.1-13.3%) were obtained. The applicability of the validated method was further verified in real Salviae miltiorrhiza Radix et Rhizoma samples.

Spontaneously removed biliary stent drainage versus T-tube drainage after laparoscopic common bile duct exploration.

Several studies have shown the safety and feasibility of laparoscopic common bile duct exploration (LCBDE) as a minimally invasive treatment options for choledocholithiasis. Use of T-tube or biliary stent drainage tube placement after laparoscopic choledochotomy for common bile duct (CBD) stones is still under debate. This study tried to confirm the safety of spontaneously removable biliary stent in the distal CBD after LCBDE to allow choledochus primary closure. A total of 47 patients with choledocholithiasis underwent LCBDE with primary closure and internal drainage using a spontaneously removable biliary stent drainage tube (stent group, N = 22) or T-tube (T-tube group, N = 25). Operative parameters and outcomes are compared. Surgical time, intraoperative blood loss, length of hospital stay, drainage tube removal time, postoperative intestinal function recovery, and cost of treatment were all significantly lower in the stent group as compared to that in the T-tube group (P < 0.05 for all). Otherwise, Bile leakage between the two groups had no significant difference (P > 0.05). The biliary stent drainage tube was excreted spontaneously 4 to 14 days after surgery with the exception of one case, where endoscopic removal of biliary tube was required due to failure of its spontaneous discharge. LCBDE with primary closure and use of spontaneously removable biliary stent drainage showed advantage over the use of traditional T-tube drainage in patients with choledocholithiasis.

Multi-walled carbon nanotubes as solid-phase extraction sorbents for simultaneous determination of type A trichothecenes in maize, wheat and rice by ultra-high performance liquid chromatography-tandem mass spectrometry.

A solid-phase extraction (SPE) procedure using multi-walled carbon nanotubes (MWCNTs) as sorbents coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed for simultaneous determination of four type A trichothecenes in maize, wheat and rice for the first time. Several key parameters including the composition of sample loading solutions, washing and elution solvents were thoroughly investigated to achieve optimal SPE recoveries and efficiency. Performance of the MWCNTs materials was significantly affected by pH, and after optimization, n-hexane and 5% methanol aqueous solution as the washing solutions and methanol containing 1% formic acid as the elution solvent presented an excellent purification efficiency for the four targets in the different matrices. The method was validated by determining the linearity (R(2)≥0.992), recovery (73.4-113.7%), precision (1.2-17.1%) and sensitivity (limit of quantification in the range of 0.02-0.10µg/kg), and was further applied for simultaneous determination of type A trichothecenes in 30 samples. Although low contamination levels of type A trichothecenes in wheat, maize and rice were observed revealing mitigated risks to consumers in Shanghai, China, the developed method has proven to be a valuable tool for type A trichothecenes monitoring in complex crop matrices.