Baicalein Relieves Ferroptosis-Mediated Phagocytosis Inhibition of Macrophages in Ovarian Endometriosis.

Iron overload and oxidative stress have been reported to contribute to ferroptosis in endometriotic lesions. However, the possible roles of iron overload on macrophages in endometriosis (EMs) remain unknown. Based on recent reports by single-cell sequencing data of endometriosis, here we found significant upregulations of ferroptosis-associated genes in the macrophage of the endometriotic lesion. Additionally, there was an elevated expression of HMOX1, FTH1, and FTL in macrophages of peritoneal fluid in EMs, as well as iron accumulation in the endometriotic lesions. Notably, cyst fluid significantly up-regulated levels of intracellular iron and ferroptosis in Phorbol-12-myristate-13-acetate (PMA)-stimulated THP-1 cells. Additionally, high iron-induced ferroptosis obviously reduced PMA-stimulated THP-1 cells' phagocytosis and increased the expression of angiogenic cytokines, such as vascular endothelial growth factor A (VEGFA) and interleukin 8 (IL8). Baicalein, a potential anti-ferroptosis compound, increased GPX4 expression, significantly inhibited ferroptosis, and restored phagocytosis of THP-1 cells in vitro. Collectively, our study reveals that ferroptosis triggered by high iron in cyst fluid promotes the development of EMs by impairing macrophage phagocytosis and producing more angiogenic cytokines (e.g., IL8 and VEGFA). Baicalein displays the potential for the treatment of EMs, especially in patients with high ferroptosis and low phagocytosis of macrophages.

[Technical recommendations for pragmatic randomized controlled trials of heat-sensitive moxibustion in community (â ¡): organization, implementation and follow-up visits].

Based on the community research experience of heat-sensitive moxibustion, this study explained technical recommendations for pragmatic randomized controlled trials (pRCTs) of heat-sensitive moxibustion in community from 7 aspects: selection of community research sites, ethical approval and registration, patient recruitment, training of standard operating procedures, ensuring patient compliance, quality control of follow-up visits and patient safety, which aimed to reduce the difficulty of research execution and improve the quality of pRCTs implementation and follow-up visits of heat-sensitive moxibustion.

Fabrication of cellulose derivative coated spherical covalent organic frameworks as chiral stationary phases for high-performance liquid chromatographic enantioseparation.

Porous spherical silica-based chiral stationary phases (CSPs) have been commercially used in the field of chiral separation, however, the scope of their application is, to some extent, limited by the instability of silica towards mobile phase containing strong base or acid. As such, developing new matrix-based CSPs is one of the effective strategies to overcome this bottleneck in studies of chiral separation materials. In this work, we have demonstrated that stable spherical covalent organic frameworks (SCOFs) can be utilized as matrixes for the fabrication of new CSPs for the first time. Specifically, a porous imine-linked SCOF with good crystallinity, large surface area, and high chemical stability is synthesized at room temperature. Then, cellulose-tris (3,5-dimethylphenylcarbamate) (CDMPC), a typical cellulose derivative, is selected as a potential chiral selector and coated onto the robust SCOFs, giving rise to the fabrication of new CDMPC@SCOF CSPs. The as-synthesized stable SCOF-based CSPs are exploited for high-performance liquid chromatographic (HPLC) enantioseparation, showing high resolution abilities for the separation of racemic compounds such as metalaxyl, 1-(1-naphthalenyl)ethanol, epoxiconazol, trans-stilbene oxide, and so on. Moreover, the prepared SCOF-based CSPs exhibit more superior acid and base stability than those of the silica-based CSPs. Our work not only uncovers the great potential of SCOFs as matrixes for constructing novel CSPs, but also expands the application of COFs in the field of enantiomeric separation under harsh base and acid conditions.

[Technical recommendation for pragmatic randomized controlled trial of heat-sensitive moxibustion in community (â ): randomization with consideration of patient preference].

Heat-sensitive moxibustion is the appropriate technique of the external treatment in traditional Chinese medicine and it is widely used in community because of its "easy learning, simple operation and clear curative effect". Pragmatic randomized controlled trial is a main intervention design in the real world study, which provides a high-level evidence for the effectiveness assessment of heat-sensitive moxibustion in community management. Focusing on the key links of randomization, e.g. block randomization, stratified randomization, cluster randomization, sample size allocation, allocation concealment and blinding, the paper elaborates the advantages, disadvantages and technical details of two-stage randomization with consideration of patient preference in pragmatic randomized controlled trials of heat-sensitive moxibustion in community. It facilitates improving the quality of evidence, reproducibility and methodological homogeneity among different trials.

Effect and Safety of Acupuncture for Type 2 Diabetes Mellitus: A Systematic Review and Meta-analysis of 21 Randomised Controlled Trials.

OBJECTIVE: To evaluate the effects of acupuncture on hypoglycaemic outcomes in type 2 diabetes mellitus (T2DM). METHODS: PubMed, Embase, Cochrane Library, and ClinicalTrials.gov were searched from inception up to July 2020, to identify randomised controlled trials (RCTs) that enrolled patients with T2DM and compared acupuncture combined with antidiabetic drugs to antidiabetic drugs alone. The primary outcomes were haemoglobin A1c (HbA1c) and fasting blood glucose (FBG). The secondary outcomes included 2-h blood glucose (2hBG), fasting insulin (FINS), homeostatic model assessment for insulin resistance (HOMA-IR), and acupuncture-related adverse events. Mean difference (MD) and 95% confidence interval (CI) were used as the effect measure in the meta-analysis. The quality of evidence was assessed using the Grading of Recommendations Assessment, Development and Evaluation tool. RESULTS: Twenty-one RCTs (n=1,188) were included. The meta-analytic results showed that the acupuncture group had greater reductions in FBG (MD -6.46 mg/dL, 95% CI -11.95 to -0.98; moderate-quality evidence) and HOMA-IR (MD -1.23, 95% CI -2.16 to -0.31; low-quality evidence), but comparable changes in HbA1c (MD -0.39%, 95% CI -0.84 to 1.61; very-low-quality evidence), 2hBG (MD -4.99 mg/dL, 95% CI -20.74 to 10.76; low-quality evidence), and FINS (MD -1.32 µIU/mL, 95% CI -3.76 to 1.12; low-quality evidence). No data on the incidence of diabetic complications were found. All acupuncture-related adverse events reported were mild. CONCLUSIONS: The current evidence suggests that acupuncture, as a complementary therapy to antidiabetic drugs, has a small but statistically significant effect on decreasing FBG and improving insulin resistance. The effects of acupuncture on HbA1c, 2hBG, and FINS remain uncertain. Acupuncture is generally safe in patients with mild diabetes. More evidence for the long-term effects of acupuncture on T2DM is needed. (Trial registration No. CRD42018115639).

An Intelligence Method for Recognizing Multiple Defects in Rail.

Ultrasonic guided waves are sensitive to many different types of defects and have been studied for defect recognition in rail. However, most fault recognition algorithms need to extract features from the time domain, frequency domain, or time-frequency domain based on experience or professional knowledge. This paper proposes a new method for identifying many different types of rail defects. The segment principal components analysis (S-PCA) is developed to extract characteristics from signals collected by sensors located at different positions. Then, the Support Vector Machine (SVM) model is used to identify different defects depending on the features extracted. Combining simulations and experiments of the rails with different kinds of defects are established to verify the effectiveness of the proposed defect identification techniques, such as crack, corrosion, and transverse crack under the shelling. There are nine channels of the excitation-reception to acquire guided wave detection signals. The results show that the defect classification accuracy rates are 96.29% and 96.15% for combining multiple signals, such as the method of single-point excitation and multi-point reception, or the method of multi-point excitation and reception at a single point.

The Co-Expression of Melanoma-Antigen Family a Proteins and New York Esophageal Squamous Cell Carcinoma-1 in Breast Cancer: A Pilot Study.

OBJECTIVE: The aim of this study was to quantify the expression of melanoma-antigen family A proteins (MAGE-A) and New York esophageal squamous cell carcinoma-1 (NY-ESO-1) in breast cancer and establish the prognosis of breast cancer patients with MAGE-A and NY-ESO-1 co-expression. METHODS: A total of 122 patients with breast cancer were recruited for this study. Their clinicopathological data were collected retrospectively, and the MAGE-A and NY-ESO-1 expressions in paraffin-embedded specimens from the 122 patients were evaluated using immunohistochemical analysis. In addition, the survival states of the patients were recorded. RESULTS: Fifty-four patients (44.26%) were MAGE-A positive and 46 (37.70%) were NY-ESO-1 positive. Interestingly, 22 of the 46 NY-ESO-1-positive cases co-expressed MAGE-A. The expression of MAGE-A was positively associated with estrogen-receptor status (χ2 = 4.026, p = 0.045) and human epidermal growth factor receptor 2 status (χ2 = 5.482, p = 0.019), while the expression of NY-ESO-1 was positively associated with p53 expression (χ2 = 4.541, p = 0.033). Of the 122 patients, the lowest survival rate was observed in patients with NY-ESO-1 (+)/MAGE-A (+), with a 5-year survival rate of 59.09% and a median survival of 97 months. CONCLUSION: The results showed that MAGE-A and NY-ESO-1 were frequently expressed in breast cancer patients. The co-expression of MAGE-A and NY-ESO-1 occurred in about 18% of these patients, and it may indicate a poor prognosis.

Ginsenosides in vascular remodeling: Cellular and molecular mechanisms of their therapeutic action.

Evidence is mounting that abnormal vascular remodeling (VR) is a vital pathological event that precedes many cardiovascular diseases (CVD). This provides us with a new research perspective that VR can be a pivotal target for CVD treatment and prevention. However, the current drugs for treating CVD do not fundamentally reverse VR and repair vascular function. The reason may be that a complicated regulatory network is formed between the various signaling pathways involved in VR. Recently, ginsenoside, the main active substance of ginseng, has become increasingly the focus of many researchers for its multiple targets, multiple pathways, and few side effects. Several data have revealed that ginsenosides can improve VR caused by vasodilation dysfunction, abnormal vascular structure and blood pressure. This review is intended to discuss the therapeutic effects and mechanisms of ginsenosides in some diseases involved in VR. Besides, we herein also give a new and contradictory insight into intracellular and molecular signaling of ginsenosides in all kinds of vascular cells. Most importantly, we also discuss the feasibility of ginsenosides Rb1/Rg1/Rg3 in drug development by combining the pharmacodynamics and pharmacokinetics of ginsenosides, and provide a pharmacological basis for the development of ginsenosides in clinical applications.

Facile and Site-Selective Synthesis of an Amine-Functionalized Covalent Organic Framework.

Amine-functionalized covalent organic frameworks (COFs) hold great potential in diversified applications. However, the synthesis is dominated by postsynthetic modification, while the de novo synthesis allowing for direct installation of amine groups remains a formidable challenge. Herein, we develop a site-selective synthetic strategy for the facile preparation of amine-functionalized hydrazone-linked COF for the first time. A new monomer 2-aminoterephthalohydrazide (NH2-Th) bearing both amine and hydrazide functionalities is designed to react with benzene-1,3,5-tricarbaldehyde (Bta). Remarkably, the different activity of amine and hydrazide groups toward aldehyde underpin the highly site-selective synthesis of an unprecedented NH2-Th-Bta COF with abundant free amine groups anchored in the well-defined pore channels. Interestingly, NH2-Th-Bta COF exhibits dramatically enhanced iodine uptake capacity (3.58 g g-1) in comparison to that of the nonfunctionalized Th-Bta COF counterpart (0.68 g g-1), and many reported porous adsorbents, despite its low specific surface area. Moreover, NH2-Th-Bta COF possesses exceptional cycling capability and retained high iodine uptake, even after six cycles. This work not only provides a simple and straightforward route for the de novo synthesis of amine-functionalized COFs but also uncovers the great potential of amine-functionalized COFs as adsorbents in the efficient removal of radioiodine and beyond.

Extraction Optimization, Physicochemical Characteristics, and Antioxidant Activities of Polysaccharides from Kiwifruit (Actinidia chinensis Planch.).

In order to evaluate effects of extraction techniques on the physicochemical characteristics and antioxidant activities of kiwifruit polysaccharides (KPS), and further explore KPS as functional food ingredients, both microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE) were optimized for the extraction of KPS. Furthermore, the physicochemical structures and antioxidant activities of KPS extracted by different techniques were investigated. The optimal extraction conditions of UAE and MAE for the extraction of KPS were obtained by response surface methodology. Different extraction techniques significantly affected the contents of uronic acids, molecular weights, molar ratios of constituent monosaccharides, and the degree of esterification of KPS. Results showed that KPS exhibited remarkable DPPH and ABTS radical scavenging activities, and reducing power. The high antioxidant activities observed in KPS extracted by the MAE method (KPS-M) might be partially attributed to its low molecular weight and high content of unmethylated galacturonic acid. Results suggested that the MAE method could be a good potential technique for the extraction of KPS with high antioxidant activity, and KPS could be further explored as functional food ingredients.

Dynamic changes in DNA demethylation in the tree shrew (Tupaia belangeri chinensis) brain during postnatal development and aging.

Brain development and aging are associated with alterations in multiple epigenetic systems, including DNA methylation and demethylation patterns. Here, we observed that the levels of the 5-hydroxymethylcytosine (5hmC) ten-eleven translocation (TET) enzyme-mediated active DNA demethylation products were dynamically changed and involved in postnatal brain development and aging in tree shrews (Tupaia belangeri chinensis). The levels of 5hmC in multiple anatomic structures showed a gradual increase throughout postnatal development, whereas a significant decrease in 5hmC was found in several brain regions in aged tree shrews, including in the prefrontal cortex and hippocampus, but not the cerebellum. Active changes in Tet mRNA levels indicated that TET2 and TET3 predominantly contributed to the changes in 5hmC levels. Our findings provide new insight into the dynamic changes in 5hmC levels in tree shrew brains during postnatal development and aging processes.

Neuroprotective properties of Madecassoside from Centella asiatica after hypoxic-ischemic injury.

Madecassoside is one of increasingly used constituent of Centella asiatica, a frequently prescribed crude drug in South eastern Asia and China for wound healing. In the present experiment, it exposes the neuroprotective nature of Madecassoside in GT1-7 cell lines, further, which the antioxidant activities are performed. The cellular toxicity was assessed using 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay with increased cell viability with IC50 2.5µg/ml. the regulation of antioxidant levels showed changes in madecassoside treated cell lysate viz., SOD assay. Also, the antioxidative assays confirmed the negligible cellular damage caused to the GT1-7 cell lines. Hence, the results advocate that the current antioxidant and antitumor activity be justified by the high concentration of phenolic constituents, primarily the triterpene present in the C. asiatica.

[Establishment of Duplex PCR for Identifying Metagonimus yokogawai and Haplorchis taichui].

OBJECTIVE: To develop a duplex PCR method for identifying Metagonimus yokogawai and Haplorchis taichui. METHODS: ITS1 sequences of M. yokogawai and H. taichui, as well as those of their homologous species were obtained from GenBank, and two sets of specific primer pairs for M. yokogawai and H. taichui were designed accordingly using Primer Premier 5.0 software. PCR reaction system and conditions were optimized. The established duplex PCR method was applied in a pool of M. yokogawai, H. taichui, and 17 related species to examine its specificity. Sensitivity was evaluated through serial dilutions of plasmids containing their specific sequences. Finally, the duplex PCR was applied to identify M. yokogawai and H. taichui among trematodes collected from the viscera of 47 cats and 40 dogs to test its practicality. RESULTS: The duplex PCR method amplified target sequences of M. yokogawai and H. taichui, generating 648 bp and 279 bp products, respectively. No cross reaction was found with the following 17 related species: Haplorchis pumilio, Clonorchis sinensis, Pharyngostomum cordatum, the metacercaria of Metorchis sp. and Exorchis sp., Echinochasmus liliputanus, Echinochasmus perfoliatus, Echinostoma friedi, Hypoderaeum conoideum, Holostephanus sp., Diplodiscus sp., Anisakis sp., Metorchis orientais, Paragonimus westermani, Watsonius watsoni, Notocotylus sp. and Hysterothylacium sp, indicating a high specificity of this method. The detection limits for DNAs of M. yokogawai and H. taichui were 1.49 x 10(-1) pg and 1.14 x 10(-1) pg, suggesting a good sensitivity for this method. Further, the duplex PCR successfully identified M. yokogawai and H. taichui from cat and dog viscera, with no cross amplification of other trematodes. CONCLUSION: The duplex PCR is effective in identifying Metagonimus yokogawai and Haplorchis taichui.

[A head-to-head comparison of contemporary indolent prostate cancer screen protocols in Chinese].

OBJECTIVE: To compare the diagnostic accuracy of five internationally used indolent prostate cancer screen protocols in Chinese prostate cancer patients. METHODS: Retrospective analysis was made of the consecutive cohort of 314 patients, from Jan. 2006 to Apr. 2014, who had both prostate biopsy and radical prostatectomy in Peking University First Hospital. The Gleason score≤6, pT2, tumor volume≤0.5 mL, margin negative and lymph nodes negative were defined as indolent prostate cancer. The predictive value of five indolent screen criteria including Epstein, Memorial Sloan-Kettering Cancer Center (MSKCC), Prostate Cancer Research International: Active Surveillance (PRIAS), University of California, San Francisco (UCSF), and University of Miami (UM) were evaluated in Chinese prostate cancer patients. Measures of diagnostic accuracy and areas under the receiver-operating curve (AUC) were calculated for each protocol and compared. RESULTS: A total of 16% (49 cases) of the patients met the inclusion criteria of at least one protocol, including 24 cases in Epstein, 33 cases in MSKCC, 28 cases in PRIAS, 34 cases in UCSF, and 22 cases in UM. Three percent were eligible for all the studied criteria. UCSF and MSKCC protocols had the highest sensitivity and specificity than the others. The Epstein and PRIAS protocols demonstrated acceptable positive predictive value, but the specificity and sensitivity were inefficient. The UM protocol was performed unsatisfiedly on sensitivity, positive predictive value and AUC. A strict limited protocol which contained all the five protocols could not improve the predictive accuracy. CONCLUSION: The UCSF protocol had better diagnostic accuracy than the others, but the results were not satisfied. A further investigation on indolent prostate cancer screening in Chinese patients is needed.

Inhibition of phosphoinositide 3-kinase delta attenuates experimental autoimmune encephalomyelitis in mice.

The phosphoinositide 3-kinase delta (PI3Kδ) has been implicated in multiple signaling pathways involved in autoimmune diseases. We here aimed to test the hypothesis that selective inhibition of PI3Kδ may promote anti-inflammatory effects by inhibiting Th1 and Th17 cells. We investigated the therapeutic efficacy of a selective PI3Kδ inhibitor IC87114 in experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis (MS). The efficacy was evaluated based on clinical scores, histopathology, serum cytokines and inflammatory infiltrations in the central nervous system (CNS). Treatment of EAE mice with IC87114 reduced the clinical symptoms, histopathology and cellular infiltration into the CNS. And treatment of EAE with IC87114 suppressed the Th1 and Th17 cell ratios. Consistently, the serum levels of IL-1ß, IL-6, IL-17 and INF-γ were markedly reduced by IC87114. Taken together, our studies demonstrate that inhibition of PI3Kδ may serve as novel therapy to suppress neuroinflammation seen during EAE.

Protective effect of SGK1 in rat hippocampal neurons subjected to ischemia reperfusion.

BACKGROUND/AIMS: To investigate the protective effect of SGK1 (serum- and glucocorticoid-inducible protein kinase 1) in rat hippocampal neurons in vitro and in vivo following ischemia reperfusion (I/R). METHODS: Isolated rat hippocampal neurons were subjected to 2 h of oxygen and glucose deprivation (OGD) then returned to normoxic conditions for 10, 30 or 60 min. Cell apoptosis and protein expression of SGK1 were analyzed. To examine SGK1 function, we overexpressed SGK1 in rat hippocampal neurons. Finally we examined the involvement of PI3K/Akt/GSK3ß signaling by treating the cells (untransfected or transfected with expression vector encoding SGK1) with the PI3K inhibitor LY294002. Findings were confirmed in vivo in a rat model of middle cerebral artery occlusion. RESULTS: I/R caused a time-dependent increase in apoptosis, both in vitro and in vivo. SGK1 protein levels decreased significantly under the same conditions. Overexpression of SGK1 reduced apoptosis following OGD or I/R compared to cells transfected with empty vector and subjected to the same treatment, or sham-operated animals. Addition of LY294002 revealed that the action of SGK1 in suppressing apoptosis was mediated by the PI3K/Akt/GSK3ß pathway. CONCLUSION: SGK1 plays a protective role in ischemia reperfusion in rat hippocampal neurons, exerting its effects via the PI3K/Akt/GSK3ß pathway.

Health status, intention to seek health examination, and participation in health education among taxi drivers in jinan, china.

BACKGROUND: Taxi drivers are exposed to various risk factors such as work overload, stress, an irregular diet, and a sedentary lifestyle, which make these individuals vulnerable to many diseases. This study was designed to assess the health status of this occupational group. OBJECTIVES: The objective was to explore the health status, the intention to seek health examination, and participation in health education among taxi drivers in Jinan, China. PATIENTS AND METHODS: The sample-size was determined scientifically. The systematic sampling procedure was used for selecting the sample. Four hundred taxi drivers were randomly selected from several taxi companies in Jinan. In total, 396 valid questionnaires (from 370 males and 26 females) were returned. Health status, intention to seek health examination, and participation in health education were assessed by a self-designed questionnaire. Other personal information including sex, age, ethnicity, marital status, years of employment as a taxi driver, education level, and habits were also collected. RESULTS: This survey revealed that 54.8% of taxi drivers reported illness in the last two weeks and 44.7% of participants reported chronic diseases. The prevalence rates of hypertension, diabetes mellitus, gastroenteritis, arthritis, and heart disease were 18.2%, 8.8%, 26%, 18.4%, and 4.8% of questioned taxi drivers, respectively. Significant self-reported symptoms included fatigue, waist and back pain, headache, dyspepsia, and dry throat affecting 49.7%, 26.2%, 23.5%, 26%, and 27% of participants, respectively. In total, 90.1% of subjects thought that it was necessary to receive a regular health examination. Only 17.9% of subjects had been given information about health education, and significantly, more than 87% of subjects who had been given information about health education reported that the information had been helpful. CONCLUSIONS: Taxi drivers' health was poor in our survey. Thus, using health education interventions to improve knowledge and change in behaviors are necessary and effective programs that improve the health of individuals in this special occupational group are needed.

Involvement of pituitary tumor transforming gene 1 in psoriasis, seborrheic keratosis, and skin tumors.

Accumulating evidence suggests that pituitary tumor transforming gene 1 (PTTG1) is a potential biomarker for cancer malignancy and a cell-cycle regulatory protein. This investigation was performed to address the subcellular localization of PTTG1 and its possible involvement in proliferative skin diseases. In vitro primary-cultured keratinocytes and skin samples from psoriasis, seborrheic keratosis (SK), basal cell carcinoma (BCC), and squamous cell carcinoma (SCC) were investigated by immunofluorescence and real-time PCR. In normal skin, PTTG1 is localized predominantly in 10% of basal keratinocytes, while 30-40% in basal and suprabasal psoriatic keratinocytes. PTTG1 mRNA in psoriatic epidermis is about 5-fold more than that in normal one (P<0.01). PTTG1 is localized in cytoplasm in primary-cultured normal and psoriatic keratinocytes, and PTTG1 in HaCaT cells is distributed throughout the cytoplasm of metaphase cells. PTTG1 is seen at both G2 and M phases, and highest PTTG1 expression correlates with highest cyclin B1 expression and highest degree of nuclear pleomorphism at M phase. The positive rate of PTTG1 in SK, BCC, and SCC is about 10%, 20%, and more than 80%, respectively. PTTG1 siRNA, which knocks down the expression of PTTG1, reduced the invasive capacity of A431 cells. In conclusion, PTTG1 is a marker for proliferative skin diseases associated with cell cycle regulation and may aid in detection of aggressive cancers.

Simultaneous quantitative determination of multiple mycotoxins in cereal and feedstuff samples by a suspension array immunoassay.

Mycotoxins produced by different species of fungi may coexist in single cereal and feedstuff samples, which could become highly toxic for humans and animals. In order to quantify four mycotoxins (zearalenone, fumonisin B1, deoxynivalenol, and aflatoxin B1) in cereal and feedstuff samples simultaneously, a new suspension array immunoassay was developed. Antimycotoxin monoclonal antibodies were conjugated to the surface of different encoding microspheres (19#, 37#, 39#, and 49#), and mycotoxin-protein conjugates were then coupled with biotin. Using streptavidin-phycoerythrin as a signal reporter protein, this direct competition multiple suspension array immunoassay was optimized. The results showed that the detection limits for zearalenone, fumonisin B1, deoxynivalenol, and aflatoxin B1 were 0.51, 6.0, 4.3, and 0.56 ng/mL, respectively, with detection ranges of 0.73-6.8, 11.6-110.3, 8.6-108.1, and 1.1-14.1 ng/mL, respectively. For the detection of the spiked samples, the recovery rates were between 92.3% and 115.5%. This method also shows a good correlation coefficient (r = 0.99, P < 0.01) with liquid chromatography-tandem mass spectrometry in the detection of toxins in commercial cereal and feedstuff samples. This suspension array immunoassay was high-throughput and accurate for the rapid quantitative detection of multiple mycotoxins in commercial cereal and feedstuff samples.

Rapid simultaneous quantification of zearalenone and fumonisin B1 in corn and wheat by lateral flow dual immunoassay.

A lateral flow dual immunoassay (LFDIA) was developed for rapid quantitative detection of zearalenone (ZEN) and fumonisin B1 (FB1) in corn and wheat samples on a single test strip. Two test lines and the control line on the nitrocellulose membrane were coated with ZEN and FB1 conjugates and goat anti-mouse IgG, respectively. Colloidal gold nanoparticles were conjugated with monoclonal antibodies against ZEN or FB1. The intensity of the test lines was analyzed by a photometric strip reader to determine the concentrations of ZEN and FB1 based on the calibration curves of known concentrations versus intensity readings. Test parameters such as types of buffers, ratio of the two gold-labeled antibodies, and dilution of the sample extracts and the gold-labeled antibodies were optimized. The detection limit was 0.35 and 5.23 ng/mL for ZEN and FB1, respectively, and the corresponding detection ranges were 0.94-7.52 and 9.34-100.45 ng/mL, respectively. Spiked and natural samples were analyzed using both LFDIA and liquid chromatography-tandem mass spectrometry. The two methods had a good correlation (R(2) = 0.96). The dual quantitative LFDIA is sensitive, rapid, and easy-to-use for on-site testing of a large number of samples.