Effect of Phenol and Alkylamide Interaction on α-Glucosidase Inhibition and Cellular Antioxidant Activity during In Vitro Digestion: Using Szechuan Pepper (Zanthoxylum genus) as a Model.

Although recent evidence indicated significant phenol and alkylamide interaction in aqueous solutions, the gastrointestinal digestion influence of the combination remains unclear. This study aims to investigate phenol and alkylamide interaction during in vitro digestion, focusing on bioaccessibility and bioactivity, including α-glucosidase inhibition and cellular antioxidant activity. Additionally, the structural mechanism of phenol and alkylamide interaction during in vitro digestion was explored. The results indicated that the presence of phenols and alkylamides significantly increased or decreased their respective bioaccessibility, depending on the Zanthoxylum varieties. Furthermore, although antagonistic phenol/alkylamide interaction was evident during α-glucosidase inhibition, cellular oxidative stress alleviation, and antioxidant gene transcription upregulation, this effect weakened gradually as digestion progressed. Glycoside bond cleavage and the methylation of phenols as well as alkylamide isomerization and addition were observed during digestion, modifying the hydrogen bonding sites and interaction behavior. This study provided insights into the phenol/alkylamide interaction in the gastrointestinal tract.

Effect of Enoxaparin and Daikenchuto Coadministration on Hepatic Disorder Markers in Gynecological Cancer Patients after Abdominal Surgery.

Enoxaparin and daikenchuto are commonly administered to prevent venous thromboembolism and intestinal obstruction after gynecological malignancy surgery. However, the effects of their combined use on hepatic function are not well studied. This study aimed to clarify the effects of the coadministration of enoxaparin and daikenchuto on hepatic function. First, Japanese Adverse Drug Event Report (JADER) data were analyzed to identify signals of hepatic disorders. Second, a retrospective observational study of patients who underwent surgery for gynecological malignancies was conducted. This study defined hepatic disorders as an increase in aspartate aminotransferase (AST) or alanine aminotransaminase (ALT) levels above the reference values, using 1-h postoperative values as the baseline. The analysis of JADER data revealed an increased risk for hepatic disorders with the coadministration of enoxaparin and daikenchuto. An observational study also showed higher odds ratios (95% confidence intervals) for the occurrence of hepatic disorders in the coadministration group (4.27; 2.11-8.64) and enoxaparin alone group (2.48; 1.31-4.69) than in the daikenchuto alone group. The median increase in the ALT level was also higher in the coadministration group (34; 15-59) than in the enoxaparin alone (19; 6-38) and daikenchuto alone groups (8; 3-33). In conclusion, our study suggests that compared with the use of enoxaparin or daikenchuto alone, enoxaparin and daikenchuto coadministration increases the risk of hepatic disorders, with more significant increases in AST and ALT levels. Healthcare workers need to be aware of these potential side effects when combining these drugs after surgery for gynecological malignancies.

Chemical Composition Variation in Essential Oil and Their Correlation with Climate Factors in Chinese Prickly Ash Peels (Zanthoxylum armatum DC.) from Different Habitats.

Essential oils are secondary metabolites in plants with a variety of biological activities. The flavor and quality of Zanthoxylum armatum DC. are mainly determined by the essential oil components in the Chinese prickly ash peels. In this study, the correlation between climate change in different regions and the content of essential oils of Z. armatum was investigated using gas chromatography-mass spectrometry (GC/MS) and multivariate statistical analysis. The Z1-24 refers to 24 batches of samples from different habitats. A total of 145 essential oils were detected in 24 batches of samples, with the highest number of terpene species and the highest content of alcohol. The relative odor activity (ROAV) values identified nine main flavor compounds affecting the odor of Z. armatum. Linalool, decanal, and d-limonene were the most critical main flavor compounds, giving Z. armatum a spicy, floral, oily, and fruity odor. The results of hierarchical cluster analysis (HCA) and principal component analysis (PCA) classified Z5 into a separate group, Z2 and Z7 were clustered into one group, and the rest of the samples were classified into another group. Correlation analysis and path analysis showed that temperature and precipitation were the main climatic factors affecting essential oils. Comparisons can be made with other plants in the genus Zanthoxylum to analyze differences in essential oil type and content. This study contributes to the identification of Z. armatum quality, promotes the accumulation of theories on the effects of climatic factors on essential oils, and enriches the site selection and breeding of Z. armatum under similar climatic conditions.

Enhancing the antioxidant properties and compatibility of protein/sodium alginate film by incorporating Zanthoxylum bungeanum essential oil Pickering emulsion.

Zanthoxylum bungeanum essential oil (ZBEO) Pickering emulsion was incorporated into rice protein (RP)/sodium alginate (SA)-based film to enhance the antioxidant activity and compatibility. With increasing ZBEO content from 2 % to 4 %, the average size of ZBEO Pickering emulsion ranged from 124.28 to 165.65 nm. The best mechanical property with a tensile strength of 14.56 MPa and hydrophobicity with a water vapor permeability of 2.11 × 10-12 g⋅cm-1⋅s-1⋅Pa-1 of emulsion film were achieved with 0.8 % ZBEO. In addition, the loss of ZBEO in the emulsion films was reduced by 11-14 %. The DPPH radical scavenging activity of emulsion film with 1.2 % ZBEO was 65.54 % in 95 % ethanol. The results of Fourier transform infrared spectroscopy and molecular dynamics simulation showed that electrostatic interactions played a leading role in film formation. Overall, ZBEO Pickering emulsion is an effective method to enhance the antioxidant activity, mechanical strength and hydrophobicity of RP/SA film.

Polyphenolic Platform Ameliorated Sanshool for Skin Photoprotection.

Natural evolution has nurtured a series of active molecules that play vital roles in physiological systems, but their further applications have been severely limited by rapid deactivation, short cycle time, and potential toxicity after isolation. For instance, the instability of structures and properties has greatly descended when sanshool is derived from Zanthoxylum xanthoxylum. Herein, natural polyphenols are employed to boost the key properties of sanshool by fabricating a series of nanoparticles (NPs). The intracellular evaluation and in vivo animal model are conducted to demonstrate the decreased photodamage score and skin-fold thickness of prepared NPs, which can be attributed to the better biocompatibility, improved free radical scavenging, down-regulated apoptosis ratios, and reduced DNA double-strand breaks compared to naked sanshool. This work proposes a novel strategy to boost the key properties of naturally occurring active molecules with the assistance of natural polyphenol-based platforms.

Research advances of Zanthoxylum bungeanum Maxim. polyphenols in inflammatory diseases.

Zanthoxylum bungeanum Maxim., commonly known as Chinese prickly ash, is a well-known spice and traditional Chinese medicine ingredient with a rich history of use in treating inflammatory conditions. This review provides a comprehensive overview of the botanical classification, traditional applications, and anti-inflammatory effects of Z. bungeanum, with a specific focus on its polyphenolic components. These polyphenols have exhibited considerable promise, as evidenced by preclinical studies in animal models, suggesting their therapeutic potential in human inflammatory diseases such as ulcerative colitis, arthritis, asthma, chronic obstructive pulmonary disease, cardiovascular disease, and neurodegenerative conditions. This positions them as a promising class of natural compounds with the potential to enhance human well-being. However, further research is necessary to fully elucidate their mechanisms of action and develop safe and effective therapeutic applications.

Chitosan/oxidized Konjac Glucomannan films incorporated with Zanthoxylum Bungeanum essential oil: A novel approach for extending the shelf life of meat.

In this study, a novel edible composite film was prepared by chitosan, konjac glucomannan oxidized with ozone for 60 min (OKGM), and Zanthoxylum Bungeanum essential oil (ZEO). The chitosan/OKGM film was fortified with ZEO to assess the physical properties, structure, antioxidant and antibacterial abilities, and pork preservation systematically. Compared to the control group, the addition of 1 % ZEO increased tensile strength by 18.92 % and decreased water solubility, water vapor permeability, and moisture content by 10.05 %, 6.60 %, and 1.03 %, respectively. However, the treatment with ZEO (1.5 % and 2 %) decreased mechanical properties and increased the water vapor permeability. The ultraviolet barrier, antioxidant, and antibacterial abilities of composite films were enhanced by increasing the ZEO addition. Moreover, the COZ-1 film was used to protect the freshness of pork with slow-release behavior of ZEO. The results showed that addition of ZEO significantly decreased the pH value, total viable count, redness, total volatile basic nitrogen, and thiobarbituric acid and increased the hardness of pork after preservation for 10 days. Therefore, the chitosan/OKGM loaded with ZEO film can potentially be used as food packaging, providing new ideas for the research on active packaging materials.

Application of chitosan films incorporated with Zanthoxylum limonella essential oil for extending shelf life of pork.

This study aimed to produce chitosan films incorporated with Zanthoxylum limonella essential oil for extending shelf life. The volatile compounds of Z. limonella essential oil were identified by gas chromatography-mass spectrometry consisting of limonene, α-phellandrene, ρ-cymene, and sabinene as major compounds. In this study, the addition of Z. limonella essential oil at concentrations of 0 %, 2 %, and 4 % in chitosan film was assessed for its antibacterial activity against Escherichia coli and Staphylococcus aureus. Chitosan film incorporated with 4 % essential oil demonstrated the most significant antibacterial effect against E. coli and S. aureus in comparison to the chitosan film without essential oil due to the synergistic effects on antibacterial activity. The physical and mechanical properties of the chitosan films incorporated with Z. limonella oil developed were also assessed. The addition of essential oil to chitosan films led to improvements in mechanical strength and flexibility, while minimal changes were observed in terms of water solubility, water vapor permeability, and thermal stability. The findings emphasize that this innovative film not only extends the shelf life of pork without chemical preservatives but is also a fully bio-based material. Consequently, it shows great potential to be used as active packaging within the food industry.

The Zanthoxylum armatum fruit's oil exterminates Candida cells by inhibiting ergosterol biosynthesis without generating reactive oxygen species.

Candida spp. is a significant cause of topical and fungal infections in humans. In addition to Candida albicans, many non-albicans species such as C. krusei, C. glabrata, C. parapsilosis, C. tropicalis, C. guilliermondii cause severe infections. The main antifungal agents belong to three different classes, including azoles, polyenes, and echinocandins. However, resistance to all three categories of drugs has been reported. Therefore, there is an urgent need to search for other alternatives with antifungal activity. Many herbal extracts and compounds from natural sources show excellent antifungal activity. In this study, we used an oil extract from the fruits of Zanthoxylum armatum, which showed significant antifungal activity against various Candida spp. by two different methods-minimum inhibitory concentration (MIC) and agar diffusion. In addition, we attempted to explore the possible mechanism of action in C. albicans. It was found that the antifungal activity of Z. armatum oil is fungicidal and involves a decrease in the level of ergosterol in the cell membrane. The decrease in ergosterol level resulted in increased passive diffusion of a fluorescent molecule, rhodamine6G, across the plasma membrane, indicating increased membrane fluidity. The oil-treated cells showed decreased germ tube formation, an important indicator of C. albicans' virulence. The fungal cells also exhibited decreased attachment to the buccal epithelium, the first step toward invasion, biofilm formation, and damage to oral epithelial cells. Interestingly, unlike most antifungal agents, in which the generation of reactive oxygen species is responsible for killing, no significant effect was observed in the present study.

Rapid determination of geographical authenticity and pungency intensity of the red Sichuan pepper (Zanthoxylum bungeanum) using differential pulse voltammetry and machine learning algorithms.

The development of an analytical method for assessing pungency intensity and determining geographical origins is crucial for evaluating the quality of visually similar Zanthoxylum bungeanum pericarp (PZB). This study analyzed 210 PZB samples from 14 origins across China, focusing on origin adulteration identification and pungency intensity using a combination of differential pulse voltammetry (DPV) and machine learning algorithms. The artificial neural network (ANN) and K-nearest neighbor (KNN) algorithms provided the highest accuracy in origin identification (100 %) and adulteration detection (97.9 %) respectively. Moreover, the ANN excelled in predicting pungency intensity (R2 = 0.918). Assessment via feature importance analysis of DPV features revealed that segments of polyphenols (0.34-0.52 V and 1.0-1.2 V) and alkylamides (1.0-1.2 V) contributed significantly to the PZB pungency intensity. These findings highlight the potential of DPV as a reliable method for assessing the quality of PZB, and offer a promising solution for ensuring the geographical authenticity of this important crop.

Effect of emulsifiers on the properties of corn starch films incorporated with Zanthoxylum bungeanum essential oil.

The use of natural and safe ingredients in green food packaging material is a hot research topic. This study investigated the effect of different emulsifiers on starch film properties. Three types of emulsifiers, including Tween 80 as a small-molecule surfactant, sodium caseinate (CAS), whey protein isolate (WPI), and gelatin (GE) as macromolecule emulsifiers, whey protein isolate fibril (WPIF) as a particle emulsifier, were utilized to prepare Zanthoxylum bungeanum essential oil (ZBO) emulsions. The mechanical, physical, thermal, antibacterial properties, microstructure and essential oil release of starch films were investigated. CAS-ZBO nanoemulsion exhibited the smallest particle size of 198.6 ± 2.2 nm. The film properties changed with different emulsifiers. CAS-ZBO film showed the highest tensile strength value. CAS-ZBO and WPIF-ZBO films exhibited lower water vapor permeability than Tween-ZBO. CAS-ZBO film showed good dispersion of essential oil, the slowest release rate of essential oils in all food simulants, and the best antibacterial effect against Staphylococcus aureus and Listeria monocytogenes. The films composed of CAS-ZBO nanoemulsion, corn starch, and glycerol are considered more suitable for food packaging. This work indicated that natural macromolecule emulsifiers of CAS and WPIF are expected to be used in green food packaging material to offer better film properties.

Neurotoxicity study of ethyl acetate extract of Zanthoxylum armatum DC. on SH-SY5Y based on ROS mediated mitochondrial apoptosis pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Zanthoxylum armatum DC. (ZADC) is a traditional medicinal plant with various pharmacological activities and is widely used in China, Japan, India, and other regions. Previous studies have revealed that the methanol extract of ZADC can cause neurotoxicity symptoms in rats, such as drooling, decreased appetite, decreased movement, and increased respiratory rate. However, the basis of these toxic substances and the mechanism of neurotoxicity remain unclear. AIM OF THE STUDY: To evaluate the effects of ZADC on nerve cells and their damage mechanisms and discuss the possible toxic substance basis. MATERIALS AND METHODS: The ethyl acetate extract of ZADC is obtained by extracting the methanol extract of ZADC with ethyl acetate. The Q-Orbitrap LC-MS/MS method was employed to analyze the chemical composition of the EA extract of ZADC. SH-SY5Y cells were incubated with different concentrations of the ethyl acetate extract of ZADC. The cytotoxicity of the extract was evaluated using CCK-8, LDH, and ROS assays, and the oxidative stress status of cells was assessed using MDA, GSH, and SOD. Cell apoptosis was detected using flow cytometry. Damage to mitochondrial function was evaluated by labeling mitochondria, ATP, and MMP with fluorescence. Cyto-C, Caspase-3, Caspase-9, Apaf-1, Bax, and reduced Bcl2 expression were measured to evaluate the activation of the mitochondrial apoptosis pathway. Finally, NAC intervention was used to detect changes in the relevant indicators. The activation of mitochondrial apoptosis pathway was evaluated by measuring Cyto-C, Caspase-3, Caspase-9, Apaf-1, and Bax and Bcl2 expression. Finally, NAC intervention was utilized to detect changes in the relevant indicators. RESULTS: After treating SY-SY5Y cells with EA extract from ZADC, cell viability decreased significantly, and the intracellular ROS level increased in a dose-dependent manner. Meanwhile, ZADC can cause cellular oxidative stress and increase MDA and SOD concentrations while decreasing GSH concentrations. It can also shorten the mitochondrial cristae and decrease the number of mitochondria. In contrast, it can reduce ATP synthesis in the mitochondria and mitochondrial membrane potential (MMP). Furthermore, it increased the apoptosis rate and the expression of Cyto-C, Caspase-3, Caspase-9, Apaf-1, and Bax and reduced Bcl2 expression. NAC intervention alleviated the reduction in SH-SY5Y cell survival and the accumulation of reactive oxygen species induced by the EA extract in ZADC. It also inhibits signaling pathways dominated by proteins, such as Cyto-C, reducing cell apoptosis and cytotoxicity. A total of 46 compounds were identified in the extracts. CONCLUSIONS: The results suggest that EA extract of ZADC can induce the mitochondrial apoptotic pathway by accumulating ROS in cells, leading to apoptosis. Antioxidants had a good inhibitory and protective effect against cell damage caused by the EA extract of ZADC. The neurotoxic components of ZADC may be organic acids and compounds containing amino groups.

Soy protein isolate-based active films functionalized with Zanthoxylum bungeanum by-products: Effects on barrier, mechanical, antioxidant and cherry tomato preservation performance.

In this work, soy protein isolate (SPI)-based films enriched with naturally sourced Zanthoxylum bungeanum leaf extract (ZBLE) were prepared. Different ZBLE contents (0, 1, 3, 5, and 7 % w/w SPI) were incorporated into the SPI matrix to investigate the effect of ZBLE on various properties of the obtained films. ZBLE exhibited excellent compatibility with SPI in terms of tensile strength, water barrier properties, UV-light resistance capability, and antioxidant activities. The films with 5 % ZBLE addition presented the most comprehensive performance. The release of total phenolic compounds in two different aqueous food simulants was analyzed. Furthermore, the films were employed to preserve fresh cherry tomatoes at 25 ± 1 °C for 18 days. The changes in the physicochemical properties (mass loss rate, decay rate, and vitamin C content) of cherry tomatoes revealed that the addition of ZBLE to films significantly extended the storage time. Therefore, the SPI/ZBLE composite film has the potential as an eco-friendly active packaging material for food preservation.

Transcriptomics integrated with metabolomics to characterize key pigment compounds and genes related to anthocyanin biosynthesis in Zanthoxylum bungeanum peel.

Zanthoxylum bungeanum is an important condiment with high economic value and its peel color is one of the main quality indexes. However, the key pigment compounds and related genes are still unclear affecting the quality control of the plants. In this study, the contents of four types of pigments were measured in Z. bungeanum and flavonoids were identified as the most important pigments. Based on the targeted flavonoid metabolomics of Z. bungeanum peels, 14 key pigment compounds were screened out from 152 flavonoids, among which cyanidin-3-O-rutinoside and cyanidin-3-O-glucoside were the most critical compounds for peel color. They were further verified to be present in nine varieties of Z. bungeanum by HPLC fingerprints. The 14 compounds were all associated with flavonoid and anthocyanin biosynthesis pathways and the 39 differentially expressed genes related to these pathways were annotated and screened based on transcriptomics. The genes ZbDFR, ZbANS, and ZbUFGT were identified as three key genes for anthocyanin synthesis in Z. bungeanum peels. Further qRT-PCR results confirmed the reliability of transcriptomics and the accuracy of gene screening. Subsequent protein induced expression demonstrated that ZbANS and ZbUFGT were expressed after 12 h induced by IPTG while ZbDFR was expressed after 15 h. Further transient and stable transformation analysis confirmed that both anthocyanin content and the expression of ZbDFR were significantly increased in overexpression Z. bungeanum leaves and Nicotiana benthamiana. The functional effect of stable transformation of ZbDFR was more significant than that of transient transformation with a 7.67-fold/1.49-fold difference in total anthocyanin content and a 42.37-fold/12.32-fold difference in the expression of ZbDFR. This study provides new insights into the chemical composition and the molecular mechanisms of Z. bungeanum peel color and lays an effective foundation for the color quality control, multi-purpose utilization of Z. bungeanum and the creation of new germplasm.

Phytochemical Analysis by FTIR of Zanthoxylum Acanthopodium, DC Fruit Ethanol Extract, N-hexan, Ethyl Acetate and Water Fraction.

Background: Andaliman fruit is used as spice in Batak cuisine, North Sumatera, Indonesia. The potency of andaliman fruit extract as herbal medicine is widely studied. Many studies elaborate the benefits of andaliman fruit extract as an antioxidant, antibacterial or anticancer. Objective: The aim of this study was to identify the phytochemical compounds of andaliman fruit extract and its fractions. Methods: The andaliman fruit was originated from Balige city, Tobasa Regency, North Sumatera. The extract was made by maceration within ethanol and followed by fractionation with n-hexan, ethyl acetate and water. The phytochemical screening by chemical reactions, thin layer chromatography, and Fourier Transform Infrared Spectrophotometer were performed. Results: This study found andaliman fruit ethanol extract consists of alkaloids, flavonoids, glycosides, tannins, triterpens, and steroids. The n-hexan fraction consists of triterpens and steroids, ethyl acetate fraction consists of flavonoids and glycosides, and water fraction consists of alkaloid, flavonoid, and glycosides. Andaliman fruit ethanol extract had eight color spots, n-hexan and ethyl acetate fraction had five color spots and water fraction had two color spots. Andaliman fruit ethanol extract had functional group of C-H alkanes, =C-H alkenes (aliphatic), O-H, C=C alkenes, C=C aromatics, C≡C alkynes, C-O, C=O, and NO2. The n-hexan fraction had C-H alkanes, =C-H alkenes (aliphatic), O-H, C=C alkenes, C-O, C=O, and NO2. The ethyl acetate fraction had C-H alkanes, =C-H alkenes (aliphatic), O-H, C=C alkenes, C=C aromatics, C-O, C=O, and NO2. The water fraction had C-H alkanes, =C-H alkenes (aliphatic), O-H, C=C aromatics, C≡C alkynes, and C-O. Conclusion: Phytochemical screening found that andaliman fruit ethanol extract, n-hexan, ethyl acetate and water fraction positive of phytochemical compounds. The FTIR of andaliman fruit ethanol extract, n-hexan fraction, ethyl acetate fraction, and water fraction showed the absorbance of C-H alkanes, O-H, C-O, and C=C alkenes indicating the presence of alkaloids, flavonoids, and triterpenoid saponin.

Optimization of enzyme-assisted microwave extraction of Zanthoxylum limonella essential oil using response surface methodology.

Zanthoxylum limonella essential oil possesses potential antimicrobial activity and is of considerable interest as food flavouring and traditional herb. In this study, an enzymolysis-pretreatment-microwave-assisted extraction (EP-MAE) method was used to extract Z. limonella essential oil. The response surface methodology (RSM) with Plackett-Burman design (PBD) and Box-Behnken design (BBD) models were employed to optimize conditions in the EP-MAE method. Seven variables including water to plant ratio, enzyme amount, incubation temperature, incubation time, shaking speed, microwave time, and microwave power were selected to determine the optimal values for extracting Z. limonella essential oil. As the results, four variables including water to plant ratio, enzyme amount, microwave time and power were evaluated as significant variables affecting on yield and volatile compounds of Z. limonella essential oil from both PBD and BBD experiments. The optimum conditions of EP-MAE was obtained as follows: water to plant ratio (11.16 mL/g), enzyme amount (0.68%), microwave time (36.73 min), and power (1665 W). The Z. limonella essential oil composition and its yield from EP-MAE was compared to those extracted from MAE and hydrodistillation. The optimal extraction conditions in the EP-MAE method enhanced significantly higher essential oil yield (7.89 ± 0.08 mg/g) compared to those found by MAE (7.26 ± 0.04 mg/g) and hydrodistillation (7.04 ± 0.03 mg/g), respectively. Fifty-one volatile components were identified among these methods, with similar major compounds of limonene, ß-pinene, and α-phellandrene, showing percentage ranging between 34.59-35.78%, 19.91-22.67%, 8.47-8.75%, respectively. However, an extremely higher content of compounds was detected using the EP-MAE method. This study demonstrates the significance of EP-MAE, which may be applied as a more potent extraction method for essential oils in aromatic plants compared to MAE and hydrodistillation.

Cytotoxicity and Genotoxicity Evaluation of Zanthoxylum rhoifolium Lam and In Silico Studies of Its Alkaloids.

The alkaloids isolated from Zanthoxylum rhoifolium have demonstrated great pharmacological potential; however, the toxic profiles of these extracts and fractions are still not well elucidated. This study evaluated the toxicity of the ethanol extract (EEZR) and neutral (FNZR) and alkaloid (FAZR) fractions. Chemical characterization was performed by chromatographic methods: thin-layer chromatography (TLC) and high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). The cytotoxicity of the samples was evaluated in human hepatocellular carcinoma (HepG2) cells using the cell viability method (MTT) and mutagenicity by the Allium cepa assay (ACA). Alkaloids isolated from the species were selected for toxicity prediction using preADMET and PROTOX. The molecular docking of the topoisomerase II protein (TOPOII) was used to investigate the mechanism of cell damage. In the EEZR, FNZR, and FAZR, the presence of alkaloids was detected in TCL and HPLC-DAD analyses. These samples showed a 50% inhibitory concentration (IC50) greater than 400 µg/mL in HepG2 cells. In ACA, time- and concentration-dependent changes were observed, with a significant reduction in the mitotic index and an increase in chromosomal aberrations for all samples. Nuclear sprouts and a micronucleus of the positive control (PC) were observed at 10 µg/mL and in the FAZR at 30 µg/mL; a chromosomal bridge in FNZR was observed at 105 µg/mL, CP at a concentration of 40 µg/mL, and nuclear bud and mitotic abnormalities in the EEZR were observed at 170 µg/mL. The alkaloids with a benzophenanthridine were selected for the in silico study, as structural alterations demonstrated certain toxic effects. Molecular docking with topo II demonstrated that all alkaloids bind to the protein. In summary, the fractionation of Z. rhoifolium did not interfere with toxicity; it seems that alkaloids with a benzophenanthridine nucleus may be involved in this toxicity.

Karyotype and Phylogenetic Relationship Analysis of Five Varieties and Cultivars of Zanthoxylum armatum Based on Oligo-FISH.

Green prickly ash (Zanthoxylum armatum) has edible and medicinal value and is an economically significant plant in many countries. Z. armatum has many cultivars and varieties with similar phenotypes that are difficult to distinguish via traditional methods. In this study, we utilized oligo-FISH to distinguish five varieties and cultivars of Z. armatum on the basis of three oligonucleotide probes of 5S rDNA, (AG3T3)3, and (GAA)6. Karyotype analysis of the five varieties and cultivars of Z. armatum showed that the Z. armatum 'Tengjiao' karyotype formula was 2n = 2x = 98m with karyotype type 1C and an arm ratio of 4.3237, including two pairs of 5S rDNA signals and five pairs of (GAA)6 signals. The karyotype formula of Z. armatum 'Youkangtengjiao' was 2n = 2x = 128m + 8sm with karyotype type 2B and an arm ratio of 3.5336, including three pairs of 5S rDNA signals and 17 pairs of (GAA)6 signals. The karyotype formula of Z. armatum var. novemfolius was 2n = 2x = 134m + 2sm with karyotype type 1C and an arm ratio of 5.5224, including two pairs of 5S rDNA signals and eight pairs of (GAA)6 signals. The karyotype formula of Z. armatum 'YT-03' was 2n = 2x = 2M + 128m + 4sm + 2st with karyotype type 2C and an arm ratio of 4.1829, including three pairs of 5S rDNA signals and nine pairs of (GAA)6 signals. The karyotype formula of Z. armatum 'YT-06' was 2n = 2x = 126m + 10sm with cytotype 2B and an arm ratio of 3.3011, including three pairs of 5S rDNA signals and two pairs of (GAA)6 signals. The five varieties and cultivars of Z. armatum had (AG3T3)3 signals on all chromosomes. The chromosomal symmetry of Z. armatum 'Tengjiao' was high, whereas the chromosomal symmetry of Z. armatum 'YT-03' was low, with the karyotypes of the five materials showing a trend toward polyploid evolution. The phylogenetic relationship between Z. armatum 'Tengjiao' and Z. armatum var. novemfolius was the closest, while that between Z. armatum 'YT-03' and Z. armatum 'YT-06' was closer than with Z. armatum 'Youkangtengjiao' according to oligo-FISH. The results provided a karyotype profile and a physical map that contributes to the distinction of varieties and cultivars of Z. armatum and provides strategies for distinguishing other cultivated species.

Ethylene-induced improvement in photosynthetic performance of Zanthoxylum armatum under reoxygenation conditions.

In this study, we evaluated the photosynthetic performance of Zanthoxylum armatum seedlings to test the tolerance to reoxygenation after waterlogging. The experiment included a control group without waterlogging (NW) and three reoxygenation groups with reoxygenation after 1day (WR1), 2days (WR2) and 3days (WR3). Seedlings were pretreated with concentrations of 0, 200 and 400µmolL-1 of ethylene. The results showed that reoxygenation after waterlogging for 1-3days decreased photosynthetic pigments content, enzymes activity, stomatal conductance (G s ), net photosynthetic rate (P n ), transpiration rate (T r ) and water-use efficiency (WUE). However, pretreatment with ethylene increased photosynthetic pigments content, enzymes activity and gas exchange parameters under both NW and WR3 treatments. The chlorophyll fluorescence results showed that the maximum quantum yield of PSII (F v /F m ) and actual photochemical efficiency of PSII (Φ PSII ) remained no significant changes under the NW and WR1 treatments, while they were significantly reduced with an increase in waterlogging days followed by reoxygenation under WR2 and WR3 treatments. Exogenous ethylene inhibited F v /F m and the non-photochemical quenching coefficient (NPQ), while enhanced Φ PSII and electron transfer efficiency (ETR) under WR2 treatments. Moreover, the accumulation of exogenous ethylene reduced photosynthetic ability. These findings provide insights into the role of ethylene in enhancing the tolerance of Z. armatum to reoxygenation stress, which could help mitigate the impact of continued climate change.

Chemical Composition, Antimicrobial, Nitric Oxide Inhibition and Cytotoxic Activity of Essential Oils from Zanthoxylum acanthopodium DC. Leaves and Stems from Vietnam.

This study was aimed to investigate the chemical composition and biological activities of leaf and stem essential oils of Zanthoxylum acanthopodium DC. from Vietnam. Their chemical composition was analyzed by GC/MS. Antimicrobial activities were evaluated by microdilution broth assay. Anti-inflammatory activity was evaluated by the ability to inhibit nitric oxide production in macrophage cells. Cytotoxic activity was evaluated using the sulforhodamine B assay on three human cancer cell lines. Forty-four compounds were identified in the leaf oil, among which dehydroaromadendrane (23.4 %), (E)-carpacin (17.6 %), 2-tridecanone (12.2 %), and 9-methyl-2-decanone (11.8 %) were the most abundant. The stem oil contained fifty-five identified constituents, mainly γ-gurjunene (51.1 %) and butyl acetate (11.8 %). Both oils exhibited inhibitory effects on three bacterial strains, namely S. aureus, E. coli, P. aeruginosa and a fungal strain C. albican, while showed insignificant effects on B. subtilis, L. fermentum, and S. enterica. Both oils showed weak NO production inhibition in LPS-induced RAW264.7 cells, but exhibited potent cytotoxic activity against all three tested cell lines SK-LU-1, MCF-7, and HepG2 with the IC50 values ranging from 16.03±0.77 to 35.60±1.62 µg/mL. This is the first report on the antimicrobial, anti-inflammatory and cytotoxic activities of essential oils from the leaves and stems of Z. acanthopodium.