•Turmeric essential oil (EO) exhibited antifungal activity both in vitro and in vivo.•Turmeric EO could suppress mycelial growth and spore germination in vitro.•Turmeric EO could disrupt fungal ...plasma membrane and mitochondrial functions.•Turmeric EO could down-regulate aflatoxin gene expression.•Turmeric EO could inhibit fungal contamination in maize in practice.
The antifungal activity and potential mechanisms in vitro as well as anti-aflatoxigenic efficiency in vivo of natural essential oil (EO) derived from turmeric (Curcuma longa L.) against Aspergillus flavus was intensively investigated. Based on the previous chemical characterization of turmeric EO by gas chromatography–mass spectrometry, the substantially antifungal activities of turmeric EO on the mycelial growth, spore germination and aflatoxin production were observed in a dose-dependent manner. Furthermore, these antifungal effects were related to the disruption of fungal cell endomembrane system including the plasma membrane and mitochondria, specifically i.e. the inhibition of ergosterol synthesis, mitochondrial ATPase, malate dehydrogenase, and succinate dehydrogenase activities. Moreover, the down-regulation profiles of turmeric EO on the relative expression of mycotoxin genes in aflatoxin biosynthetic pathway revealed its anti-aflatoxigenic mechanism. Finally, the suppression effect of fungal contamination in maize indicated that turmeric EO has potential as an eco-friendly antifungal agent.
•Thirty-one organophosphorus pesticides residues in Alpinia oxyphylla were determined.•Sample preparation was completed in a single step without any clean-up procedure.•The proposed method was ...successfully applied to 55 commercial samples.•Seven organophosphorus pesticides were detected in eight of the samples.•The positive samples were further confirmed by GC–MS/MS.
A simple, rapid and effective gas chromatography–flame photometric detection method was established for simultaneous multi-component determination of 31 organophosphorus pesticides (OPPs) residues in Alpinia oxyphylla, which is widely consumed as a traditional medicine and food in China. Sample preparation was completed in a single step without any clean-up procedure. All pesticides expressed good linear relationships between 0.004 and 1.0μg/mL with correlation coefficients higher than 0.9973. The method gave satisfactory recoveries for most pesticides. The limits of detection varied from 1 to 10ng/mL, and the limits of quantification (LOQs) were between 4 and 30ng/mL. The proposed method was successfully applied to 55 commercial samples purchased from five different areas. Five pesticide residues were detected in four (7.27%) samples. The positive samples were confirmed by gas chromatography with tandem mass spectrometry (GC–MS/MS).
Until now, despite the great success acquired in scientific research and commercial applications, magnetic beads (MBs) have been used for nothing more than a carrier in most cases in bioassays. In ...this work, highly chemiluminescent magnetic beads containing N-(4-aminobutyl)-N-ethyl isoluminol (ABEI) and Co2+ (Co2+/ABEI/MBs) were first synthesized via a facile strategy. ABEI and Co2+ were grafted onto the surface of carboxylated MBs by virtue of a carboxyl group and electrostatic interaction. The as-prepared Co2+/ABEI/MBs exhibited good paramagnetic properties, satisfactory stability, and intense chemiluminescence (CL) emission when reacted with H2O2, which was more than 150 times that of ABEI functionalized MBs. Furthermore, it was found that 2,4,6-trinitrotoluene (TNT) aptamer could attach to the surface of Co2+/ABEI/MBs via electrostatic interaction and coordination interaction between TNT aptamer and Co2+, leading to a decrease in CL intensity due to the catalytic site Co2+ being blocked by the aptamer. In the presence of TNT, TNT would bind strongly with TNT aptamer and detach from the surface of Co2+/ABEI/MBs, resulting in partial restoration of the CL signal. Accordingly, label-free aptasensor was developed for the determination of TNT in the range of 0.05–25 ng/mL with a detection limit of 17 pg/mL. This work demonstrates that Co2+/ABEI/MBs are easily connected with recognition biomolecules, which are not only magnetic carriers but also direct sensing interfaces with excellent CL activity. It provides a novel CL interface with a magnetic property which easily separates analytes from the sample matrix to construct label-free bioassays.
•Toxigenic fungi were recovered in functional foods and spices.•Contamination levels of mycotoxins were determined by HPLC-FLD.•Aspergillus and Penicillium were the predominant fungi.•Some samples ...were found to be positive with mycotoxins, especially AFB1.
Twenty-four samples including 14 functional foods and 10 spices obtained from Chinese markets were examined for their mould profile. The mycotoxin contamination levels were also determined by an optimized HPLC-FLD method. 124 fungal isolates belonging to four different genera were recovered with Aspergillus and Penicillium as predominant fungi, with an incidence of 66.1% and 15.3%, respectively. In functional foods Aspergillus niger section (57.1%) was isolated more frequently, followed by Aspergillus flavi section (50.0%) and Aspergillus ochraceus section (21.4%), with the most contaminated samples being Coix seeds. Similar fungal presence and frequency were encountered in spice with A. niger section group (60.0%) and A. flavi section (40.0%) as main fungi. Cumin and Pricklyash peel samples showed the highest fungal contamination. Four functional foods and three spices were found to be positive at low levels for mycotoxins including aflatoxin B1 (up to 0.26μg/kg) and ochratoxin A (OTA) (5.0μg/kg). The more frequently detected mycotoxin was AFB1 (16.7%).
Vancomycin, which has a strong antibacterial effect to Gram-positive bacteria, was adopted as one molecular recognition agent for bacterial detection. Magnetic beads (MBs) were functionalized with ...this antibiotic to effectively concentrate Staphylococcus aureus (S. aureus). In addition, alkaline phosphatase (ALP)-tagged rabbit immunoglobulin G (ALP-IgG) was used as the second recognition agent to improve the specificity based on the binding between the Fc region of rabbit IgG and protein A in the cell wall of S. aureus. MBs-concentrated sandwich complex of vancomycin/S. aureus/ALP-IgG was formed with a one-step incubation protocol. Then ALP chemiluminescent reaction was triggered by injecting substrate solution to quantitate S. aureus. Based on the sandwich molecular recognition mechanism and MBs concentration, an ultrasensitive, specific and rapid method was developed for S. aureus detection. The linear range for S. aureus detection was 12–1.2×106CFUmL−1, with a very low detection limit of 3.3CFUmL−1. The whole detection process could be completed in 75min. Other Gram-positive bacteria and Gram-negative bacteria, including Escherichia coli, Salmonella, Pseudomonas aeruginosa, Micrococcus luteus, Bacillus cereus and Bacillus subtilis, showed negligible interference to S. aureus detection. This method was successfully used to quantitate S. aureus in lake water, milk, human urine and human saliva with acceptable recoveries ranging from 70.0% to 116.7%.
•An ultrasensitive, specific and rapid CL method was developed for S. aureus detection.•The method was based on the sandwich molecular recognition and MBs concentration.•The method is believed to show an acceptable specificity.
Identifying plant, fungal, and animal ingredients in a specific mixture remains challenging during the limitation of PCR amplification and low specificity of traditional methods. Genomic DNA was ...extracted from mock and pharmaceutical samples. Four type of DNA barcodes were generated from shotgun sequencing dataset with the help of a local bioinformatic pipeline. Taxa of each barcode was assigned by blast to TCM-BOL, BOLD, and GenBank. Traditional methods including microscopy, thin layer chromatography (TLC), and high-performance liquid chromatography (HPLC) were carried out according to Chinese pharmacopoeia. On average, 6.8 Gb shotgun reads were sequenced from genomic DNA of each sample. Then, 97, 11, 10, 14, and one operational taxonomic unit (OTU) were generated for ITS2, psbA-trnH, rbcL, matK, and COI, respectively. All the labeled ingredients including eight plant, one fungal, and one animal species were successfully detected in both the mock and pharmaceutical samples, in which Chebulae Fructus, Poria, and Fritilariae Thunbergia Bulbus were identified via mapping reads to organelle genomes. In addition, four unlabeled plant species were detected from pharmaceutical samples, while 30 genera of fungi, such as Schwanniomyces, Diaporthe, Fusarium were detected from mock and pharmaceutical samples. Furthermore, the microscopic, TLC, and HPLC analysis were all in accordance with the standards stipulated by Chinese Pharmacopoeia. This study indicated that shotgun metabarcoding could simultaneously identified plant, fungal, and animal ingredients in herbal products, which has the ability to serve as a valuable complement to traditional methods.
•A widely applicable strategy for aflatoxins analysis in medicinal herbs was proposed.•The critical points of purification approach was optimized.•The method was validated by a certified reference ...material and representative matrices.•The method was successfully used to determine the aflatoxin level of real samples.
Although extraction methods based on immunoaffinity column (IAC) cleanup have been used to detect aflatoxins in medicinal herbs, they do not yield satisfactory results for all sample matrices. The difficulty arises from the chemical complexity of the herbs, and there is a pressing need to determine which steps in IAC cleanup limit the scope of aflatoxin detection in many different kinds of medicinal herbs. In this work, we found that there were two main factors that severely decreased antibody-antigen recognition and led to serious nonspecific adsorption: (1) high extract acidity and (2) high co-extraction of interfering compounds. We therefore carried out a systematic study to optimize extraction efficiency. We found that dilution of samples in 0.1M phosphate buffer solution (pH 7.8, 2% Tween-20) at a 1:8 dilution ratio mitigated the effect of high acidity, decreased co-precipitation of compounds and nonspecific adsorption, and ameliorated the matrix effect. To validate this finding, and test if our method is widely applicable to in different kinds of herbal materials, we analyzed several representative complex sample matrices including fructus, cortex, and radix with varying extract pH values. The recovery efficiency was generally higher than 70%. We further validated our method by testing a certified reference material, and found that our approach accurately quantified aflatoxin concentration. After validation, we successfully used this method to determine the aflatoxin concentration of real samples. The approach described here could potentially be used to extract aflatoxin from other complex matrices with varying acidity.
Anterior cervical discectomy and fusion (ACDF) is a common procedure for treating cervical spondylotic myelopathy (CSM), however, ACDF may cause pseudoarthrosis, accelerated degeneration of adjacent ...segments, loss of activity of fused segments and other complications. The full-endoscopic technique can treat CSM, without the aforementioned complications above. Therefore, it is of great clinical value to investigate the surgical scheme of anterior percutaneous full-endoscopic transcorporeal decompression of the spinal cord (APFETDSC).
A total of 28 cases with single-segment Cervical spondylotic myelopathy (CSM) from April 2017 to July 2019 were involved in this study. The size of the disc-osteophyte complex was measured using imaging data prior to the operation. The diameter and direction of the bony passage was determined according to the size and central position of the complex, respectively. Twenty-eight patients underwent the above scheme for CSM. The clinical outcome evaluations included Visual Analog Scale (VAS) scores, Japanese Orthopedic Association (JOA) scores. The imaging assessment included MRI, CT and X-rays.
The diameter of the designed bony passage was about 6.9 mm, and directed toward the lower edge of the diseased lower vertebral body oblique to the center of the disc-osteophyte complex. All patients successfully completed the operation. The postoperative neck pain VAS and JOA were significantly improved compared to preoperative values (p < 0.01). Postoperative MRI indicated complete decompression of the spinal cord. CT scanning 1 year after the operation revealed an almost healed bony passage and X-ray imaging showed satisfactory physiological curvature of the cervical spine, without cervical instability.
Based on the diameter and direction of the bony passage, as determined by the size and position of the disc-osteophyte complex, indicated by MRI and CT scanning, anterior percutaneous full-endoscopic transcorporeal decompression of the spinal cord offers good decompression of the spinal cord and ensures excellent therapeutic outcome.
The freshwater leech Whitmania pigra (W. pigra) Whitman (Annelida phylum) is a model organism for neurodevelopmental studies. However, molecular biology research on its embryonic development is still ...scarce. Here, we described a series of developmental stages of the W. pigra embryos and defined five broad stages of embryogenesis: cleavage stages, blastocyst stage, gastrula stage, organogenesis and refinement, juvenile. We obtained a total of 239.64 Gb transcriptome data of eight representative developmental phases of embryos (from blastocyst stage to maturity), which was then assembled into 21,482 unigenes according to our reference genome sequenced by single-molecule real-time (SMRT) long-read sequencing. We found 3114 genes differentially expressed during the eight phases with phase-specific expression pattern. Using a comprehensive transcriptome dataset, we demonstrated that 57, 49 and 77 DEGs were respectively related to morphogenesis, signal pathways and neurogenesis. 49 DEGs related to signal pathways included 30 wnt genes, 14 notch genes, and 5 hedgehog genes. In particular, we found a cluster consisting of 7 genes related to signal pathways as well as synapses, which were essential for regulating embryonic development. Eight genes cooperatively participated in regulating neurogenesis. Our results reveal the whole picture of W. pigra development mechanism from the perspective of transcriptome and provide new clues for organogenesis and neurodevelopmental studies of Annelida species.
