Cold plasma for enhancing covalent conjugation of ovalbumin-gallic acid and its functional properties.

The utilization of cold plasma (CP) treatment to promote covalent conjugation of ovalbumin (OVA) and gallic acid (GA), as well as its functionality, were investigated. Results demonstrated that CP significantly enhanced the covalent grafting of OVA and GA. The maximum conjugation of GA, 24.33 ± 2.24 mg/g, was achieved following 45 s of CP treatment. Covalent conjugation between GA and OVA were confirmed through analyses of total sulfhydryl (-SH) group, Fourier transform infrared (FTIR) spectroscopy, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Unfolding of the OVA molecule occurred upon conjugation with GA, as evidenced by multiple spectroscopy analyses. Additionally, conjugation with GA resulted in significant improvements in the antioxidant activity and emulsifying properties of OVA. This study demonstrated that CP is a robust and sustainable technique for promoting the covalent conjugate of polyphenols and proteins, offering a novel approach to enhance the functional properties of proteins.

Preventing thermal aggregation of ovalbumin through dielectric-barrier discharge plasma treatment and enhancing its emulsification properties.

The impact of Dielectric-Barrier Discharge (DBD) plasma treatment on the prevention of heat-induced aggregation of Ovalbumin (OVA) and improvement in emulsification properties was investigated. Results highlighted the effective inhibition of thermal aggregation of OVA following exposure to plasma. Structural analysis revealed that the plasma-induced oxidation of sulfhydryl and intermolecular disulfide bonds played a pivotal role in inhibiting the thermal aggregation, considered by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE), multiplies spectroscopy, and analysis of dynamic exchange of sulfhydryl-disulfide bonds. Meanwhile, the oxidation of exposed hydrophobic sites due to plasma treatment resulted in the transformation of the OVA molecule's surface from hydrophobic to hydrophilic, contributing significantly to the aggregation inhibition. Additionally, compared to an untreated sample of OVA, almost one-fold increase in emulsifying ability (EAI) and 1.5-fold in emulsifying stability (ESI) was observed after 4 min of plasma treatment. These findings demonstrated that plasma treatment not only enhanced the thermal stability of OVA, but also improved its emulsification properties.

Precise measurement of microwave polarization using a Rydberg atom-based mixer.

A Rydberg atom-based mixer has opened up a new method to characterize microwave electric fields such as the precise measurement of their phase and strength. This study further demonstrates, theoretically and experimentally, a method to accurately measure the polarization of a microwave electric field based on a Rydberg atom-based mixer. The results show that the amplitude of the beat note changes with the polarization of the microwave electric field in a period of 180 degrees, and in the linear region a polarization resolution better than 0.5 degree can be easily obtained which reaches the best level by a Rydberg atomic sensor. More interestingly, the mixer-based measurements are immune to the polarization of the light field that forms the Rydberg EIT. This method considerably simplifies theoretical analysis and the experimental system required for measuring microwave polarization using Rydberg atoms and is of interest in microwave sensing.

Laparoscopic resection of a rare diaphragmatic haemangioma: a case report.

This case report describes the laparoscopic resection of a rare diaphragmatic haemangioma. A 45-year-old male patient was diagnosed incidentally with a left subphrenic mass by computed tomography. Laparoscopic left subphrenic mass excision was performed under general anaesthesia. A phrenic haemangioma was confirmed by postoperative pathology. Tumours originating in the diaphragm are rare, with only approximately 200 cases reported in the past century. The diaphragmatic tumour was determined to be primary because intraoperative imaging showed that the tumour was relatively isolated and had no obvious relationship with the surrounding tissues and organs.

Paralytic ileus as first symptom of Miller Fisher syndrome: A case report.

INTRODUCTION: Miller Fisher syndrome (MFS), regarded by many scholars as a variant of Guillain Barre syndrome (GBS), accounts for approximately 5% to 10% of GBS cases. The typical clinical manifestations of MFS are extraocular muscle paralysis, ataxia, and tendon reflex loss or disappearance. To date, intestinal obstruction has rarely been reported as the initial symptom. PATIENT CONCERNS: A 48-year-old woman presenting with abdominal pain and distention was diagnosed with paralytic ileus. There was no significant improvement in symptoms after symptomatic treatment. After that, the patient developed visual rotation, with limited binocular abduction and adduction, and ataxia. Anti-ganglioside testing revealed positive anti-ganglioside antibodies. DIAGNOSIS: The patient was diagnosed as MFS. INTERVENTIONS: The early stage is mainly symptomatic treatment of paralytic ileus. After MFS was diagnosed, the patient was given large amounts of immunoglobulin and hormone shock therapy. OUTCOMES: After 1 week, the symptoms of intestinal obstruction and MFS gradually improved. The patient was later discharged automatically for financial reasons. Six months after discharge, the patient was interviewed by telephone, and she had recovered. CONCLUSION: To date, intestinal obstruction has rarely been reported as the initial symptom. In case of inconsistencies between the imaging examinations and clinical symptoms, neuroelectrophysiology and cerebrospinal fluid puncture should be performed, striving for timely detection and treatment.

Effects of Dietary Bopu Powder Supplementation on Serum Antioxidant Capacity, Egg Quality, and Intestinal Microbiota of Laying Hens.

The purpose of this study was to investigate the effects of dietary Bopu powder supplementation on the serum antioxidant capacity, serum biochemical indices, egg quality, and intestinal microbiota. Six hundred and forty-eight 33-week-old Lohmann Brown commercial laying hens were randomly allocated into six groups and fed a basal diet supplemented with 0, 25, 50, 100, 200, and 400 mg/kg Bopu powder for 8 weeks, denoted BP0, BP25, BP50, BP100, BP200, and BP400, respectively. The results showed that dietary Bopu powder supplementation reduced serum cholesterol concentrations (linear, p < 0.01) while increasing serum globulin and albumin concentrations (linear, p < 0.05). Furthermore, the BP50 and BP100 groups had greater serum catalase and glutathione peroxidase activity (p < 0.05). The egg Haugh Units were considerably higher in BP25 and BP50 (p < 0.05), and eggshell thickness was higher in BP25, BP200, and BP400 (p < 0.05) when compared to BP0. Dietary treatment with Bopu powder at doses ranging from 25-100 mg/kg improved glutathione peroxidase and catalase activities while decreasing malondialdehyde concentrations in the yolk (p < 0.05). The addition of Bopu powder increased the diversity of microbiota and the relative abundance of Bacteroidota in the gut. For instance, dietary Bopu powder supplementation of 25-50 mg/kg significantly raised the relative abundance of Enterococcus, Bacteroides, and Fusobacterium in the foregut. Supplementing the diet with 50-100 mg/kg of Bopu powder improved the relative abundance of Lactobacillus in the hindgut. In conclusion, dietary Bopu powder supplementation enhanced the abundance of beneficial bacteria in the foregut of laying hens and improved egg quality and antioxidant capacity. Furthermore, in the laying hen diet, the optimal dosage of Bopu powder additive was 25-50 mg/kg.

Identification of seven undescribed cucurbitacins in Cucumis sativus (cucumber) and their cytotoxic activity.

Cucurbitacin C-type cucurbitacins that are only identified in Cucumis sativus (cucumber) are, in part, responsible for the health benefits and bitter flavor. Nevertheless, detailed information about those functional ingredients in cucumber is scarce. In this study, ten cucurbitacin C analogues including seven undescribed ones have been isolated from the bitter leaves of cucumber, in which six compounds showed growth inhibition capabilities against tumor cell lines HepG2, A549, DU145 and HCT116. Intriguingly, cucurbitacin C6 and C7 exhibited a significant inhibition effect compared to the positive control taxol (IC50 = 1.86 ± 0.17 µM) on HepG2 cell line with IC50 values of 10.06 ± 0.34 µM and 4.16 ± 0.42 µM, respectively. The mechanism of cucurbitacin-induced apoptosis is likely down-regulating the expression of caspase-related proteins. This work enlarges the knowledge of the cucurbitacins in cucumber and highlights the importance of cucumber as a source of specialized metabolites in the food and medicinal industries.

Oxidation induced by dielectric barrier discharge (DBD) plasma treatment reduces IgG/IgE binding capacity and improves the functionality of glycinin.

The effect of dielectric barrier discharge (DBD) plasma treatment times from 2 to 5 min at 40 kV on IgG/IgE binding capacity and functionality of soybean glycinin was examined. A substantial reduction in the binding capacity (91.64% for IgG and 81.49% for IgE) was obtained after 5 min of plasma treatment, as determined by western-blot and ELISA analyses. Further studies demonstrated that the elimination of antigenicity and allergenicity of glycinin was directly related to plasma-induced structural changes on two aspects. A conformational alteration caused by oxidation of peptide bond amino groups, accompanied with an oxidation of Trp, Tyr, and Phe amino acid residues, which was confirmed by surface hydrophobicity, multi-spectroscopic analysis, and amino acid analysis. The cleavage of polypeptide chains inevitably partially diminished the linear epitopes, resulting in a primary decline in IgG/IgE binding capacity. Additionally, an increase in the solubility from 10.78 ± 0.35 to 65.96 ± 1.86% and significant increase in the emulsifying ability from 21.08 ± 2.64 to 160.29 ± 4.12 m2/g were observed after treatment of the plasma for 2 min. The present results confirm the potential use of DBD for the production of hypoallergenic soy protein-based products and improving their technical functions such as solubility and emulsifying ability.

The chromosome-level Hemerocallis citrina Borani genome provides new insights into the rutin biosynthesis and the lack of colchicine.

Hemerocallis citrina Borani (huang hua cai in Chinese) is an important horticultural crop whose flower buds are widely consumed as a delicious vegetable in Asia. Here we assembled a high-quality reference genome of H. citrina using single-molecule sequencing and Hi-C technologies. The genome assembly was 3.77 Gb and consisted of 3183 contigs with a contig N50 of 2.09 Mb, which were further clustered into 11 pseudochromosomes. A larger portion (3.25 Gb or 86.20%) was annotated as a repetitive content and 54,295 protein-coding genes were annotated in the genome. Genome evolution analysis showed that H. citrina experienced a recent whole-genome duplication (WGD) event at ~15.73 million years ago (Mya), which was the main factor leading to many multiple copies of orthologous genes. We used this reference genome to predict 20 genes involved in the rutin biosynthesis pathway. Moreover, our metabolomics data revealed neither colchicine nor its precursors in H. citrina, challenging the long-standing belief that this alkaloid causes poisoning by the plant. The results of our disruptive research are further substantiated by our genomic finding that H. citrina does not contain any genes involved in colchicine biosynthesis. The high-quality genome lays a solid foundation for genetic research and molecular breeding of H. citrina.

Plant-derived isoquinoline alkaloids that target ergosterol biosynthesis discovered by using a novel antifungal screening tool.

The ergosterol pathway is a prime antifungal target as it is required for fungal survival, yet is not involved in human homeostasis. Methods to study the ergosterol pathway, however, are often time-consuming. The minimum inhibitory concentration (MIC) assay is a simple research tool that determines the lowest concentration at which a novel antimicrobial is active in vitro with limited scope to determine the mechanism of action for a drug. In this study, we show that by adding hydrogen peroxide, an oxidative stressor, or glutathione (GSH), an antioxidant, to modify a commonly performed MIC assay allowed us to screen selectively for new antifungal drugs that target ergosterol biosynthesis in fungi. A human pathogen and dermatophyte, Microsporum gypseum, was used as a test organism. When exposed to ergosterol targeting drugs, the hydrogen peroxide treatment significantly decreased fungal survival by reducing ergosterol in the cell wall, whereas GSH increased survival of M. gypseum. Further, by performing a series of experiments with M. gypseum and Trichophyton rubrum, it was determined that the oxidative stress from hydrogen peroxide causes cell death at different developmental stages based on fungal species. These findings allow us to describe a simple, high-throughput method for simultaneously screening new antifungal drugs for activity and effects on the ergosterol pathway. By using this tool, two isoquinoline alkaloids were discovered to be potent inhibitors of ergosterol biosynthesis in vitro by reducing the amount of ergosterol without affecting the expression of 1,3-ß-glucan. Both compounds also significantly reduced the severity of acanthosis, hyperkeratosis, spongiosis and dermal edema in vivo.

Fermented traditional Chinese medicine alters the intestinal microbiota composition of broiler chickens.

For a long time, phytogenic resources have been widely used as substitutes for antibiotics in livestock production. However, few studies have examined the relationship between the intestinal microbiota and fermented herbal medicines. Here, 252 Arbor Acres broiler plus broiler chickens were randomly assigned to a control group, which was fed a basal diet; an unfermented healthy chicken powder group (JJS) fed a basal diet containing 20 g/kg JJS; or one of 5 fermented JJS groups, which were fed a basal diet containing 20 g/kg JJS, fermented with by 5 different bacterial strains for 42 days. The growth performances of the different groups were measured and the changes in the intestinal microbiota were analyzed. The body weight gain in the Bacillus subtilis group (Bs) was the highest among the 6 groups, while the feed conversion ratio (FCR) was best with Z. rouxii fermentation. The result indicated that products of JJS fermentation products of JJS by B. subtilis and Z. rouxii had important effects on chicken growth performance. The foregut and hindgut microbial communities of Bs, Zr, the control group and the JJS group, were collected for 16S rDNA sequencing. The results showed that JJS and its fermentation products mainly acted on the foregut and had little effect on the hindgut, and Z. rouxii fermentation products can increased the diversity in the foregut microbial community. In addition, the relative abundance of Bifidobacterium in the foregut of the Z. rouxii group was significantly increased, which may be an important factor in promoting growth.

Integrating Network Pharmacology and Molecular Docking to Analyse the Potential Mechanism of action of Macleaya cordata (Willd.) R. Br. in the Treatment of Bovine Hoof Disease.

Based on network pharmacological analysis and molecular docking techniques, the main components of M. cordata for the treatment of bovine relevant active compounds in M. cordata were searched for through previous research bases and literature databases, and then screened to identify candidate compounds based on physicochemical properties, pharmacokinetic parameters, bioavailability, and drug-like criteria. Target genes associated with hoof disease were obtained from the GeneCards database. Compound-target, compound-target-pathway-disease visualization networks, and protein-protein interaction (PPI) networks were constructed by Cytoscape. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed in R language. Molecular docking analysis was done using AutoDockTools. The visual network analysis showed that four active compounds, sanguinarine, chelerythrine, allocryptopine and protopine, were associated with the 10 target genes/proteins (SRC, MAPK3, MTOR, ESR1, PIK3CA, BCL2L1, JAK2, GSK3B, MAPK1, and AR) obtained from the screen. The enrichment analysis indicated that the cAMP, PI3K-Akt, and ErbB signaling pathways may be key signaling pathways in network pharmacology. The molecular docking results showed that sanguinarine, chelerythrine, allocryptopine, and protopine bound well to MAPK3 and JAK2. A comprehensive bioinformatics-based network topology strategy and molecular docking study has elucidated the multi-component synergistic mechanism of action of M. cordata in the treatment of bovine hoof disease, offering the possibility of developing M. cordata as a new source of drugs for hoof disease treatment.

Improvement of antifungal activity of a culture filtrate of endophytic Bacillus amyloliquefaciens isolated from kiwifruit and its effect on postharvest quality of kiwifruit.

This study reports the optimization of culture conditions and medium components of M9, an endophytic strain of Bacillus amyloliquefaciens, which we had previously isolated from kiwifruit, and evaluation of its effect on kiwifruit postharvest protection against soft rot. The method of one-factor-at-a-time was used to determine the optimum culture conditions, and response surface methodology was applied to optimize the medium constituents. After optimization, a high rate of antifungal activity (73.12% decrease in decay rate) by M9 culture filtrate against the soft rot fungal pathogen, Botryosphaeria dothidea, was obtained. Compared with the initial culture conditions, the antifungal activity of M9 culture filtrate was increased by 19.5%. Furthermore, a postharvest storage experiment on kiwifruit showed that M9 culture filtrate could maintain the quality of stored kiwifruit, delay fruit senescence, and significantly enhance the storability of kiwifruit. Using liquid chromatography-mass spectrometry, we found that the antifungal activity of M9 was associated with the presence of the C12-surfactin A lipopeptide. PRACTICAL APPLICATIONS: Soft rot caused by Botryosphaeria dothidea is one of the most important diseases of kiwifruit, causing postharvest fruit rot. The disease progresses rapidly and is difficult to control, posing a great threat to the kiwifruit industry. In this study, the culture conditions and medium composition for culture of the strain M9 (kiwifruit endophytic Bacillus amyloliquefaciens) were optimized to maximize the inhibition of B. dothidea. A postharvest storage experiment showed that M9 culture filtrate could improve the disease resistance of kiwifruits, delay the senescence of the fruits, and maintain the quality of kiwifruit during storage. Because M9 is a natural endophyte of kiwifruit, the strategy used in this study was both effective and safe. This work will contribute to the exploitation of B. amyloliquefaciens in controlling the pathogens of kiwifruit and the development of safer and more advanced strategies for the postharvest protection of kiwifruits.

Chemical Composition and Bioactivity of Essential Oil of Ten Labiatae Species.

Using antibiotics as feed additives have been successively banned worldwide from 1986; therefore, it is an urgent task to finding safe and effective alternatives. As natural products of plant origin, essential oils (EOs) are an outstanding option due to their reported bioactivity. In this research, ten EOs of Labiatae species were extracted by steam distillation and its chemical constituents were identified by gas chromatography-mass spectrometry (GC-MS). A total of 123 chemical compounds, including alkenes, phenols, aldehydes and ketones, were identified. The results of antioxidant activity carried out through DPPH free radical scavenging (DPPH) and ferric reducing antioxidant power (FRAP), showing that EOs of Ocimum basilicum Linn. (ObEO), Thymus mongolicus Ronn. (TmEO), Origanum vulgare Linn. (OvEO) and Mosla chinensis Maxim. (McEO) have strong antioxidant activities. Their 50%-inhibitory concentration (IC50) value was <1.00, 1.42, 1.47 and 1.92 µg/mL, respectively; and their FRAP value was 1536.67 ± 24.22, 271.84 ± 4.93, 633.71 ± 13.14 and 480.66 ± 29.90, respectively. The results of filter paper diffusion showing that McEO, OvEO and TmEO inhibition zone diameter (IZD) are all over 30 mm. The results of two-fold dilution method showed that McEO, OvEO and TmEO have strong antibacterial activities against Staphylococcus aureus (S. aureus) and their minimal inhibitory concentrations (MIC) value was 1 µL/mL, 2 µL/mL, and 2 µL/mL, respectively. In conclusion, the results in this work demonstrate the possibility for development and application of EOs as potential feed additives.

Dispersive microwave electrometry using Zeeman frequency modulation spectroscopy of electromagnetically induced transparency in Rydberg atoms.

We herein developed and demonstrated a Zeeman frequency modulation scheme for improving the signal-to-noise ratio of microwave electric field measurement using Rydberg atoms. The spectra of the electromagnetically induced transparency (EIT) and Autler-Townes splitting of Rydberg atoms is frequency modulated by an alternating current magnetic field. The signal-to-noise ratio of the corresponding dispersive error signal is enhanced more than 10 times than that of the original spectrum. Furthermore, we show that the slope of the dispersive error signal near the resonance of the Rydberg EIT can be used to characterize the weak microwave electric field amplitudes. The more intuitive and simpler structure compared with other existing frequency modulation technologies greatly reduces the difficulties of experiments and experimental data analysis.

Systematic identification of compounds in Macleaya microcarpa by high-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry combined with mass spectral fragmentation behavior of Macleaya alkaloids.

RATIONALE: Macleaya microcarpa (Maxim.) Fedde belongs to the genus Macleaya of the Papaveraceae family. Benzylisoquinoline alkaloids (BIAs) are considered the main bioactive constituents of M. microcarpa. METHODS: Using high-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry (HPLC/QTOFMS/MS) we identified BIAs in the aerial parts of M. microcarpa in the early flowering stage. Target profiling and identification of BIAs in the extracted samples from the fresh aerial parts of M. microcarpa were exclusively based on a personal, accurate, mass database of known compounds and the mass spectral fragmentation behavior of Macleaya alkaloids. RESULTS: A total of 97 alkaloids, comprising 7 benzyltetrahydroisoquinolines, 1aporphine, 9 tetraprotoberberines, 3 protoberberines, 2 N-methyltetrahydroprotoberberines, 4 protopines, 47 dihydrobenzophenanthridines, and 24 benzophenanthridines, were identified from the fresh aerial parts of M. microcarpa, and 77 of these were detected for the first time in M. microcarpa. In addition, some of the screened alkaloids were related to the biosynthetic pathways of sanguinarine and chelerythrine. CONCLUSIONS: The integrated method is sensitive and reliable for screening and identifying trace or ultra-trace isoquinoline alkaloids and has contributed to a better understanding of BIAs in the fresh aerial parts of M. microcarpa.

The chicken gut metagenome and the modulatory effects of plant-derived benzylisoquinoline alkaloids.

BACKGROUND: Sub-therapeutic antibiotics are widely used as growth promoters in the poultry industry; however, the resulting antibiotic resistance threatens public health. A plant-derived growth promoter, Macleaya cordata extract (MCE), with effective ingredients of benzylisoquinoline alkaloids, is a potential alternative to antibiotic growth promoters. Altered intestinal microbiota play important roles in growth promotion, but the underlying mechanism remains unknown. RESULTS: We generated 1.64 terabases of metagenomic data from 495 chicken intestinal digesta samples and constructed a comprehensive chicken gut microbial gene catalog (9.04 million genes), which is also the first gene catalog of an animal's gut microbiome that covers all intestinal compartments. Then, we identified the distinctive characteristics and temporal changes in the foregut and hindgut microbiota. Next, we assessed the impact of MCE on chickens and gut microbiota. Chickens fed with MCE had improved growth performance, and major microbial changes were confined to the foregut, with the predominant role of Lactobacillus being enhanced, and the amino acids, vitamins, and secondary bile acids biosynthesis pathways being upregulated, but lacked the accumulation of antibiotic-resistance genes. In comparison, treatment with chlortetracycline similarly enriched some biosynthesis pathways of nutrients in the foregut microbiota, but elicited an increase in antibiotic-producing bacteria and antibiotic-resistance genes. CONCLUSION: The reference gene catalog of the chicken gut microbiome is an important supplement to animal gut metagenomes. Metagenomic analysis provides insights into the growth-promoting mechanism of MCE, and underscored the importance of utilizing safe and effective growth promoters.

Hairy root induction and benzylisoquinoline alkaloid production in Macleaya cordata.

Sanguinarine is currently widely used to replace antibiotic growth promoters in animal feeding and has demonstrated useful anticancer activity. Currently, the main source of sanguinarine is from an important medicinal plant, Macleaya cordata. To obtain a new source of sanguinarine production, we established hairy root cultures of M. cordata by co-cultivating leaf and stem explants with Agrobacterium rhizogenes. Except the co-cultivation medium, all growth media contained 200 mg/L timentin to eliminate A. rhizogenes. Through comparing the metabolic profiles and gene expression of hairy roots and wild-type roots sampled at five time points, we found that the sanguinarine and dihydrosanguinarine contents of hairy roots were far higher than those of wild-type roots, and we revealed the molecular mechanism that causes these metabolites to increase. Consequently, this study demonstrated that the hairy root system has further potential for bioengineering and sustainable production of sanguinarine on a commercial scale. To the best of our knowledge, this is the first efficient protocol reported for the establishment of hairy root cultures in M. cordata using A. rhizogenes.

Characterization of in vitro metabolites of three tetrahydroprotoberberine alkaloids in rat liver S9 by high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.

RATIONALE: Tetrahydroberberine (THB), tetrahydrocoptisine (THCP) and tetrahydrocolumbamine (THCB) belong to the tetrahydroprotoberberine (THPB) alkaloids. Most of them have been extensively studied because of their pharmacological activities such as anti-hypertension, anti-arrhythmia, antimicrobial activity and antioxidant. However, limited information on the pharmacokinetics and metabolism of the three alkaloids has been reported. The purpose of this study was to investigate the in vitro metabolism of THB, THCP and THCB in rat liver S9 by using a rapid and accurate high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC/QqTOF-MS) method. METHODS: The incubation mixture was processed with 15% trichloroacetic acid. Chromatographic separation of the three THPB alkaloids and their metabolites was achieved by HPLC/QqTOF-MS and accurate mass measurements of metabolites were automatically performed through data-dependent acquisition in only a 30-min analysis. The detailed structural elucidations of these metabolites were performed by comparing the changes in their accurate molecular masses, elemental compositions and product ions with those of the parent drug. RESULTS: Five, five and four metabolites of THB, THCP and THCB were identified in rat liver S9, respectively. The results show that O-demethylenation of the 9,10-vicinal methoxyl group was the main metabolic pathway of THB and THCB and that demethylenation of the two methylenedioxy groups was the main metabolic pathway of THCP. In addition, minor oxidation and methylation reactions could occur for these alkaloids in rat liver S9. CONCLUSIONS: This was the first investigation of the in vitro metabolism of THB, THCP and THCB in rat liver S9 by using a sensitive and accurate HPLC/QqTOF-MS method. The tentatively proposed metabolic pathways of the three alkaloids will provide a basis for further studies of the in vivo metabolism of the three compounds in animals and humans.

Medicinal plants of the genus Macleaya (Macleaya cordata, Macleaya microcarpa): A review of their phytochemistry, pharmacology, and toxicology.

In the genus Macleaya, Macleaya cordata and Macleaya microcarpa have been recognized as traditional herbs that are primarily distributed in China, North America, and Europe and have a long history of medicinal usage. These herbs have been long valued and studied for detumescence, detoxification, and insecticidal effect. This review aims to provide comprehensive information on botanical, phytochemical, pharmacological, and toxicological studies on plants in the genus Macleaya. Plants from the genus of Macleaya provide a source of bioactive compounds, primarily alkaloids, with remarkable diversity and complex architectures, thereby having attracted attention from researchers. To date, 291 constituents have been identified and/or isolated from this group. These purified compounds and/or crude extract possess antitumor, anti-inflammatory, insecticidal, and antibacterial activities in addition to certain potential toxicities. Macleaya species hold potential for medicinal applications. However, despite the pharmacological studies on these plants, the mechanisms underlying the biological activities of active ingredients derived from Macleaya have not been thoroughly elucidated to date. Additionally, there is a need for research focusing on in vivo medical effects of Macleaya compounds and, eventually, for clinical trials.