Ganoderma lucidum triterpenoids investigating their role in medicinal applications and genomic protection.

OBJECTIVES: Ganoderma lucidum (GL) is a white rot fungus widely used for its pharmacological properties and health benefits. GL consists of several biological components, including polysaccharides, sterols, and triterpenoids. Triterpenoids are often found in GL in the form of lanostane-type triterpenoids with quadrilateral carbon structures. KEY FINDINGS: The study revealed that triterpenoids have diverse biological properties and can be categorized based on their functional groups. Triterpenoids derived from GL have shown potential medicinal applications. They can disrupt the cell cycle by inhibiting ß-catenin or protein kinase C activity, leading to anti-cancer, anti-inflammatory, and anti-diabetic effects. They can also reduce the production of inflammatory cytokines, thus mitigating inflammation. Additionally, triterpenoids have been found to enhance the immune system's defenses against various health conditions. They possess antioxidant, antiparasitic, anti-hyperlipidemic, and antimicrobial activities, making them suitable for pharmaceutical applications. Furthermore, triterpenoids are believed to afford radioprotection to DNA, protecting it from radiation damage. SUMMARY: This review focuses on the types of triterpenoids isolated from GL, their synthesis pathways, and their chemical structures. Additionally, it highlights the pharmacological characteristics of triterpenoids derived from GL, emphasizing their significant role in various therapeutic applications and health benefits for both humans and animals.

Unraveling the Metabolomics Mysteries in Camellia Oil: From Cognition to Application.

Camellia oil is a high-value edible seed oil, recommended by the Food and Agriculture Organization (FAO). It is essential to develop accurate and rapid analytical methods to authenticate camellia oil due to its susceptibility to adulteration. Recently, hyphenated chromatography-mass spectrometry, especially high-resolution mass spectrometry using chemometrics, has become a promising platform for the identification of camellia oil. Based on the compositional analysis, the fatty acid, sterol, phenol, and tocopherol profiles (or fingerprints) were utilized as predictor variables for assessing authenticity. The review systematically summarizes the workflow of chromatography-mass spectrometry technologies and comprehensively investigates recent metabolomic applications combined with chemometrics for camellia oil authentication. Metabolomics has significantly improved our understanding of camellia oil composition at the molecular level, contributing to its identification and full characterization. Hence, its integration with standard analytical methods is essential to enhance the tools available for public and private laboratories to assess camellia oil authenticity. Integrating metabolomics with artificial intelligence is expected to accelerate drug discovery by identifying new metabolic pathways and biomarkers, promising to revolutionize medicine.

Hyphenated chromatography-MS represents a precise method for camellia oil characterizationMetabolomics-based HRMS enhances camellia oil identification at the molecular levelMachine learning shows promise in ensuring camellia oil authenticityFatty acids, phenols, and sterols are metabolic markers for camellia oil authenticationMultivariate modeling detects camellia oil fraud through diverse data analysis.

Aflatoxin B1-induced liver pyroptosis is mediated by disturbing the gut microbial metabolites: The roles of pipecolic acid and norepinephrine.

The disturbed gut microbiota is a key factor in activating the aflatoxin B1 (AFB1)-induced liver pyroptosis by promoting inflammatory hepatic injury; however, the pathogen associated molecular pattern (PAMP) from disturbed gut microbiota and its mechanism in activating liver pyroptosis remain undefined. By transplanting AFB1-originated fecal microbiota and sterile fecal microbial metabolites filtrate, we determined the association of PAMP in AFB1-induced liver pyroptosis. Notably, AFB1-originated sterile fecal microbial metabolites filtrate were more active in triggering liver pyroptosis in mice, as compared to parental fecal microbiota. This result supported a critical role of the metabolic homeostasis of gut microbiota in AFB1-induced liver pyroptosis, rather than an injurious response to direct exposure of AFB1 in liver. Among the gut-microbial metabolites, pipecolic acid and norepinephrine were proposed to bind TLR4 and NLRP3, the upstream proteins of pyroptosis signaling pathway. Besides, the activations of TLR4 and NLRP3 were linearly correlated with the concentrations of pipecolic acid and norepinephrine in the serum of mice. In silenced expression of TLR4 and NLRP3 in HepG2 cells, pipecolic acid or norepinephrine did not able to activate hepatocyte pyroptosis. These results demonstrated the necessity of gut microbial metabolism in sustaining liver homeostasis, as well as the potential to provide new insights into targeted intervention for AFB1 hepatotoxicity.

Brigatinib combined with cetuximab in the fifth-line treatment of non-small cell lung cancer with EGFR p.C797S mutation in critically ill patients: a report of two cases and literature review.

For critically ill patients with non-small cell lung cancer (NSCLC) in need of life-saving treatment, there is currently no reported evidence regarding the use of medication specifically targeting epidermal growth factor receptor ( EGFR ) p.C797S mutation, which is known to cause resistance to third-generation tyrosine kinase inhibitors (TKIs). Our report aims to investigate and explore treatment strategies to overcome resistance associated with EGFR p.C797S mutation in order to provide potential therapeutic options for these patients. Here, we reported two cases with NSCLC who initially harbored an EGFR -sensitive mutation and were both treated with osimertinib, a third-generation TKI. Next-generation sequencing tests conducted prior to the initiation of fifth-line therapy in critically ill patients revealed the presence of EGFR p.C797S mutations in both patients, suggesting acquired resistance. In the course of fifth-line therapy, the administration of a combination of brigatinib and cetuximab proved vital in saving critically ill patients, moderately extending their overall survival period. Our findings suggested that a combined regimen of brigatinib and cetuximab could serve as a potentially life-saving therapeutic strategy for critically ill patients with NSCLC, particularly those demonstrating EGFR p.C797S-mediated resistance. Further studies, however, are required to validate and expand upon these promising findings.

Discrimination rancidity degree of infant formula rice flour based on Headspace Solid-Phase Microextraction combined with Gas Chromatography-Mass Spectrometry as an alternative to sensory evaluation.

To mitigating the serious threat of harmful volatile substances to the health of infants, an alternative method of odor evaluation were proposed based on Headspace solid-phase microextraction (HS-SPME) combined with Gas Chromatography-Mass Spectrometry (GC-MS) to discriminate the degree of rancidity of infant formula rice flour (IFRF). Inspectors can simply calculate the rancidity degree of infant formula rice flour according to the regression equation based on the concentration of rancidity markers. The results showed that the joint application of OPLS-DA, molecular sensory experiments, and unsaturated fatty acids (UFAs) degradation experiments could successfully recognize the rancidity markers without collinearity in multiple linear regression analysis. The rancidity markers curve fitting was helpful for the establishment of multivariate regression model of rancidity grading. The model had an accuracy of more than 92.90% by the verification of odor evaluation. The application of the model to investigate the market IFRF samples showed that about 3% of the samples collected in the experiment were unsuitable for infant feeding. Therefore, the established model was considered to be a robust and less workload method to replace the olfactory evaluation method for discriminating the rancidity degree of IFRF.

Based on the Cancer Genome Atlas Database Development of a prognostic model of RNA binding protein in stomach adenocarcinoma.

The purpose of this study was to identify potential RNA binding proteins associated with the survival of gastric adenocarcinoma, as well as the corresponding biological characteristics and signaling pathways of these RNA binding proteins. RNA sequencing and clinical data were obtained from the cancer genome map (N = 32, T = 375) and the comprehensive gene expression database (GSE84437, N = 433). The samples in The Cancer Genome Atlas were randomly divided into a development group and a test group. A total of 1495 RNA binding protein related genes were extracted. Using nonparametric tests to analyze the difference of RNA binding protein related genes, 296 differential RNA binding proteins were obtained, 166 were up-regulated and 130 were down regulated. Twenty prognosis-related RNA binding proteins were screened using Cox regression, including 14 high-risk genes (hazard ratio > 1.0) and 6 low-risk genes (hazard ratio < 1.0). Seven RNA binding protein related genes were screened from the final prognostic model and used to construct a new prognostic model. Using the development group and test group, the model was verified with survival analysis, receiver operating characteristics curves and prognosis analysis curves. A prediction nomogram was finally developed and showed good prediction performance.

Boosting ultralong chemiluminescence for the self-powered time-resolved immunosensor.

The construction of an immunosensor based on ultralong chemiluminescence is challenged due to the shortage of highly efficient initiator for long and stable catalysis. Herein, the heterogeneous Au/Pt@CuO/Cu2O catalyst was used to investigate the structure-activity relationship, while Au/Pt significantly promotes the activity of CuO/Cu2O to catalyze H2O2 and thus produces ·OH and O2•- radicals in highly alkaline solutions, resulting in the strong and long chemiluminescence in the reaction with luminol (10 mL, more than 4 min with 1 µg catalyst). By using the Au/Pt@CuO/Cu2O as the label in the immunoassay, the strong and long chemiluminescence could initiate the photocurrent of the photoelectrochemical (PEC) substrate, and the luminescence time could influence the photocurrent extinction time, thus a self-powered time-resolved PEC immunosensor was developed to detect furosemide, showing a linear relationship between the extinction time and the logarithm of concentrations from 10-3 to 1 µg/L. This work not only experimentally verifies that the Pt-O-Cu bond in heterogeneous catalysts breaks the pH limitation of the Fenton reaction, but also realizes the chemiluminescence for self-powered time-resolved immunosensor, thereby expanding the portable applicability of chemiluminescence in food safety inspection, health monitoring, and biomedical detection without external light source.

Nickel-Catalyzed, Enantioselective Hydrofluoromethylation of Olefins: Access to Chiral α-Fluoromethylated Amides and Esters.

We herein report the nickel-catalyzed enantioselective hydrofluoromethylation of enamides and enol esters with CH2FI as the fluoromethyl source to enable the diversity-oriented synthesis (DOS) of chiral α-fluoromethylated amides as well as esters with features of wide functional group compatibility as well as excellent enantioselectivity. The synthetic value of this protocol was demonstrated by transformations of the resulted α-fluoromethylated amides to different scaffolds including amine, oxazoline, thiazoline, and α-fluoromethylated tetrahydroquinoline.

Active film preparation using pectin and polyphenols of watermelon peel and its applications for super-chilled storage of chilled mutton.

We investigated the effect of active packaging films prepared by pectin (WMP) and polyphenols (WME) obtained from watermelon peel on the quality of chilled mutton during super-chilled storage. The addition of WME created new chemical and hydrogen bonds in film. Furthermore, an appropriate amount of WME (≤1.5%) was evenly distributed throughout the film matrix, improving barrier properties, mechanical properties, thermal stability, and light transmittance of the film. An assessment of the meat quality showed that the pH, L*, b*, thiobarbituric acid reactive substances (TBARs), total volatile basic nitrogen (TVB-N), and total bacterial count (TCA) of super-chilled + film group were significantly lower, whereas shear force and a* value were significantly higher (P < 0.05) than other groups. The WMP/WME film has dense microstructure and excellent mechanical properties after storage. Pectin and polyphenols obtained from watermelon peel have good potential as a novel packaging material for chilled mutton during super-chilled storage.

Facile Fabrication of Highly Quantum Dot/AuNP-Loaded Tags for a Dual-Modal Colorimetric/Reversed Ratiometric Fluorescence Immunochromatographic Assay.

Developing an easily-prepared, sensitive, and accurate point-of-need immunochromatographic assay (ICA) is significant in food safety screening, clinical diagnosis, and environmental monitoring. However, the current single-modal ICAs are limited in certain instinct drawbacks that restrict analytical performances. Herein, we introduce an ultrasensitive dual-modal colorimetric/reversed ratiometric fluorescence ICA based on facilely prepared immunoprobes with a high loading capacity of red quantum dots and AuNPs. By smartly integrating these red-colored/fluorescent signal probes with an immobilized green quantum dot antigen on the test lines, discrete "turn-on" visual inspection and reversed ratiometric quantification via a portable smartphone-based analyzer were accomplished. As an application, this method was employed to detect 11 phosphodiesterase-5 inhibitors in health foods with ultralow detection limits (0.0028-0.045 ng/mL), high repeatability (coefficient of variations of 0.3-1.91%), and reasonable accuracy (recoveries of 86.6-107%). The proposed method was further validated by the authorized liquid chromatography with tandem mass spectrometry method in actual sample detection. This new assay format can be extended to ultrasensitive flexible detection of other food contaminants, environmental pollutants, or tumor biomarkers within minutes, and it just requires simply prepared signal reporters, easy-to-operate procedures, and a low-cost miniaturized analyzer.

A monoclonal antibody-based time-resolved fluorescence microsphere lateral flow immunoassay for paclobutrazol detection.

Paclobutrazol (PBZ) is a plant growth inhibitor and fungicide, but it is also carcinogenic and teratogenic, and has potential harm to human health. In this study, two PBZ haptens (PBZ-1, PBZ-2) were synthesized and conjugated with carrier proteins to get artificial antigens. A highly specific monoclonal antibody (mAb) against PBZ was prepared. The antibody subtype was IgG1 and the concentration was 11.03 mg/mL. A sensitive and rapid time-resolved fluorescence microsphere lateral flow immunoassay (TRFMs-LFIA) was established based on the mAb. The activated pH, the mAbs diluents, the mAb reacting concentration and the probe amount were optimized. The visual limit of detection (vLOD) and quantitative limit of detection (qLOD) of the TRFMs-LFIA for PBZ were 50 and 1.72 ng/mL respectively, and the 50% inhibiting concentration (IC50) was 9.38 ng/mL. The pretreatment procedures are simple and rapid, and the detection time of TRFMs-LFIA strip is 6 min. Qualitative and quantitative analysis of PBZ could be achieved under a UV light or with a portable fluorescence immunoassay analyzer. The average recovery rates ranged from 96.2% to 111.9% and the corresponding coefficients of variation (CV) were 4.0%-11.2% in spiked wheat and rice samples. Twenty real wheat and rice samples were measured by the TRFMs-LFIA and compared with Ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS). The measured values showed a good accordance. These results indicated that the proposed assay will provide a novel effective strategy for on-site detection of PBZ.

Soluble Sema4D Level Is Positively Correlated with Sema4D Expression in PBMCs and Peripheral Blast Number in Acute Leukemia.

Semaphorin 4D (Sema4D) is highly expressed in various cancers and leukemia. It is involved in the development of acute leukemia. A high level of soluble Sema4D is also present in the plasma of acute leukemia patients. However, it remains unknown whether Sema4D is associated with the clinical characteristics of acute leukemia. In this study, Sema4D expression was examined in peripheral blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BMMCs) of patients with acute leukemia, and it was highly expressed in the PBMCs of B-acute lymphoblastic leukemia (ALL), T-ALL, and acute myeloid leukemia (AML) patients and in the BMMCs of B-ALL and AML patients but not in the BMMCs of T-ALL patients. Sema4D expression was higher in the PBMCs of T-ALL patients than in the PBMCs of B-ALL or AML patients. In addition, Sema4D expression in BMMCs was reduced in B-ALL patients during the chemotherapy process. It was lower in remission patients than in newly diagnosed and patients without remission. In acute leukemia, soluble Sema4D level in serum is positively correlated with Sema4D expression in PBMCs, leukocyte number, and peripheral blast number. Those results suggest that the levels of Sema4D and its soluble form are associated with acute leukemia development and may be regarded as a potential biomarker in pediatric acute leukemia.

Mimotope-Based Immunoassays for the Rapid Analysis of Mycotoxin: A Review.

Mycotoxins are toxic contaminants in foods and feeds that are naturally occurring and largely unavoidable. Determining their contents in these products is essential to protect humans from harm. Immunoassays of mycotoxins have been well-established because they are fast, sensitive, simple, and cost-effective. However, a major limitation of immunoassays is the requirement of toxic mycotoxins as competing antigens, standards, or competing tracers. Mimotopes are peptides or proteins that can specifically bind to antibodies and compete with analytes for binding sites by mimicking antigenic epitopes. They can be employed as substitutes for competing antigens, standards, or competing tracers to avoid use of mycotoxins. This review summarizes the production and functionalization of the two main kinds of mimotopes, mimic peptides and anti-idiotypic antibodies (Ab2), and their applications in rapid analysis of mycotoxins.

"Dual-Signal-On" Integrated-Type Biosensor for Portable Detection of miRNA: Cas12a-Induced Photoelectrochemistry and Fluorescence Strategy.

The abnormal expression of microRNA (miRNA) can affect the RNA transcription and protein translation, leading to tumor progression and metastasis. Currently, the accurate detection of aberrant expression of miRNA, particularly using a portable detection system, remains a great challenge. Herein, a novel dual-mode biosensor with high sensitivity and robustness for miR-21 detection was developed based on the cis-cleavage and trans-cleavage activities of Cas12a. miRNA can be combined with hairpin DNA-horseradish peroxidase anchored on a CdS/g-C3N4/B-TiO2 photoelectrode, thus the nonenzymatic amplification was triggered to form numerous HRP-modified double-stranded DNA (HRP-dsDNA). Then, HRP-dsDNA can be specifically recognized and efficiently cis-cleaved by Cas12a nucleases to detach HRP from the substrate, while the remaining HRP on HRP-dsDNA can catalyze 4-chloro-1-naphthol (4-CN) to form biocatalytic precipitation (BCP) on the surface of the photoelectrode, and thus the photocurrent can be changed. Meanwhile, the trans-cleavage ability of Cas12a was activated, and nonspecifically degrade the FQ-reporter and a significant fluorescence signal can be generated. Such two different kinds of signals with independent transmission paths can mutually support to improve the performance of the detection platform. Besides, a portable device was constructed for the point-of-care (POC) detection of miR-21. Moreover, the dual-mode detection platform can be easily expanded for the specific detection of other types of biomarkers by changing the sequence of hairpin DNA, thereby promoting the establishment of POC detection for early cancer diagnosis.

Polystyrene Microsphere-Based Immunochromatographic Assay for Detection of Aflatoxin B1 in Maize.

Aflatoxin B1 (AFB1), a mycotoxin, is hepatotoxic, carcinogenic, and nephrotoxic in humans and animals, and contaminate a wide range of maize. In this study, an immunochromatographic assay (ICA) based on polystyrene microspheres (PMs) was developed for sensitive and quantitative detection of AFB1 in maize. The amounts of PMs, the condition for activating carboxyl groups of PMs, the amount of monoclonal antibody (mAb), and the volume of the immune probe were optimized to enhance the performance PMs-ICA for point-of-care testing of AFB1 in maize. The PMs-ICA showed the cut-off value of 1 ng/mL in phosphate buffer (PB) and 6 µg/kg in maize samples, respectively. The quantitative limit of detection (qLOD) was 0.27 and 1.43 µg/kg in PB and maize samples, respectively. The accuracy and precision of the PMs-ICA were evaluated by analysis of spiked maize samples with recoveries of 96.0% to 107.6% with coefficients of variation below 10%. In addition, the reliability of PMs-ICA was confirmed by the liquid chromatography-tandem mass spectrometry method. The results indicated that the PMs-ICA could be used as a sensitive, simple, rapid point-of-care testing of AFB1 in maize.

Rational hapten design to produce high-quality antibodies against carbamate pesticides and development of immunochromatographic assays for simultaneous pesticide screening.

Carbamate pesticides (CPs) are the most used pesticides in agricultural production and pest control. In this study, carbofuran, isoprocarb and carbaryl were employed as models, and a general hapten strategy based on carbamate moiety recognition was proposed. Molecular modeling of the three-dimensional (3D) structure and surface electrostatic potential of the CPs indicated that the amide group formed by conjugation significantly influenced recognition by antibodies. The proposed strategy was used to obtain three sensitive and specific monoclonal antibodies (mAbs) with IC50 values of 1.4 ng/mL, 8.4 ng/mL and 13.8 ng/mL for carbofuran, isoprocarb and carbaryl, respectively. Negligible cross-reactivity (%) with analogs was observed, except for fenobucarb (84.6%) for isoprocarb. The obtained antibodies were used to develop an immunochromatographic assay (ICA) to simultaneously and quantitatively detect the three CPs. A strip reader was used to determine the limits of quantitation (LOQs) as 0.05 ng/mL (carbofuran), 31.3 ng/mL (isoprocarb) and 31.3 ng/mL (carbaryl). The recoveries of cucumber and Chinese cabbage samples ranged from 76% to 111%, with CVs from 1.3% to 10.6%, indicating good potential for the rapid simultaneous detection of multiple pesticide residues in a large batch of samples.

Structural characterization and transcript-metabolite correlation network of immunostimulatory effects of sulfated polysaccharides from green alga Ulva pertusa.

Three water-soluble polysaccharides (UPPs 1-3) were obtained from edible green alga Ulva pertusa. The chemico-physical analyses indicated that UPPs 1-3 possessed molecular weights of 376.7 kDa, 57.21 kDa, and 131.13 kDa, with sulfate contents of 26.01 ± 8.13%, 9.86 ± 3.24%, and 13.32 ± 6.56%, respectively, and composed of arabinose, galactose, glucose, xylose, galacturonic acid, glucuronic acid, and mannuronic acid, with different ratios. The in vitro studies revealed that UPP-1 showed significant effects on the proliferation and phagocytic activity of macrophage, release of nitric oxide, and secretion of cytokines (TNF-α and IL-6). The transcript-metabolite analysis of UPP-1 treated macrophage revealed 4747 differential genes (2416 up-regulated and 2331 down-regulated) and 94 differential metabolites (77 up-regulated and 17 down-regulated) that significantly co-mapped a transcript-metabolite correlation network of biosynthesis of amino acids, glycerophospholipid metabolism, and carbon metabolism. Thus, these findings provide a valuable foundation for the potential application of U. pertusa polysaccharides.

Development of Cu(II)/Cu(I)-induced quantum dot-mediated fluorescence immunoassay for the sensitive determination of ethyl carbamate.

A series of haptens were rationally designed for producing monoclonal antibodies specific for EC and a simple fluorescence immunoassay platform was developed for the sensitive determination of EC based on alkaline phosphatase (ALP)-triggered Cu+ quenching of CdSe quantum dots (QDs). It was noted that Cd as a fluorescence substrate in CdSe QDs can be selectively substituted by Cu+ that resulted in a more significant fluorescence quenching in comparison with Cu2+. Meanwhile, because ALP catalyzed ascorbic acid phosphate and then assisted the transformation of Cu2+ to Cu+, the change in fluorescence intensity was found to be proportional to ALP concentration. After simple magnetic separation, the sensitivity and linear range of the established assay were improved approximately 53-fold and an order of magnitude, respectively, when compared with the conventional ELISA. The proposed platform was able to both amplify the signal and eliminate matrix interferences, making it a promising to determine EC as well as other contaminants in complex food matrix in a highly sensitive and simple manner. Graphical abstract.

Ultrasensitive Fluorometric Angling Determination of Staphylococcus aureus in Vitro and Fluorescence Imaging in Vivo Using Carbon Dots with Full-Color Emission.

Rapid, accurate, and safe screening of foodborne pathogenic bacteria is essential to effectively control and prevent outbreaks of foodborne illness. Fluorescent sensors constructed from carbon dots (CDs) and nanomaterial-based quenchers have provided an innovative method for screening of pathogenic bacteria. Herein, an ultrasensitive magnetic fluorescence aptasensor was designed for separation and detection of Staphylococcus aureus (S. aureus). Multicolor fluorescent CDs with a long fluorescent lifetime (6.73 ns) and high fluorescence stability were synthesized using a facile hydrothermal approach and modified cDNA as a highly sensitive fluorescent probe. CD fluorescence was quenched by Fe3O4 + aptamer via fluorescence resonance energy transfer (FRET). Under optimal conditions, the FRET-based aptasensor can detect S. aureus accompanied by a wide linear range of 50-107 CFU·mL-1 and a detection limit of 8 CFU·mL-1. Compared with other standard methods, this method was faster and more convenient, and the entire test was finished within 30 min. The capability of the aptasensor was simultaneously investigated on food samples. Additionally, the developed CDs exhibited excellent biocompatibility and were thus applied as fluorescent probes for bioimaging both in vitro and in vivo. This new platform provided an excellent application of the CDs for detecting and bioimaging pathogenic bacteria.