First Report of Alternaria alternata Causing Leaf blight on Epimedium sagittatum (Sieb. et Zucc.) Maxim. in China.

Epimedium sagittatum (Sieb.et Zucc.) Maxim. is an important material of traditional Chinese medicine because of the rich content of flavonoids that are used to treat osteoporosis, liver cancer, and sexual dysfunction (Liu et al. 2013). A leaf blight was observed on E. sagittatum in Zhumadian City, China (32°58'12" N, 114°37'48" E, continental monsoon climate) in June 2021. Survey indicated that about 18% of the plants were infected in a 266-ha commercial planting area. The initial symptoms were white patches with tan borders, irregular in outline, with small black particles visible on the center of the lesions. In a week or so, patches extended throughout the leaf, and then leaves withered. Thirty leaves with symptoms collected from five different sites were cut into 5×5 mm pieces, and then surface-sterilized with 75% ethanol for 15 s followed by rinsing with double distilled water (ddH2O) three times. The pieces were then disinfested with 0.1% HgCl2 solution for 30 s, and rinsed with ddH2O, then placed onto potato-dextrose agar medium (PDA) and incubated in the dark for 3 d at 28°C. Eight fungal isolates were purified; of these, only the isolate HY2-1 infected the host plant and was selected for further morphological characterization. The colonies of HY2-1 were olive green with loose aerial hyphae on PDA. Conidiophores were single or branched, producing brown conidia in short chains. Conidia were obclavate, obpyriform, or ellipsoidal, 15.9-47.3 µm × 7.6-16.6 µm (n=50) and pale brown or dark brown with a short cylindrical beak at the tip that contained 1-5 transverse septa and 0-4 longitudinal septa. Morphological characteristics of the isolate were identical with those of Alternaria species (Huang et al. 2022). For molecular identification, the internal transcribed spacers (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Weir et al. 2012), major allergen Alt a 1(Alt a 1) and translation elongation factor 1-α gene (TEF) (Lawrence et al. 2013) were amplified and sequenced using the primers ITS4/5, GDF/GDR, Alt-F/R, and EF1-728F/986R, respectively. The results of the sequencing were uploaded to GenBank as ITS (OR418487), GAPDH (OR419792), Alt a 1 (OR419794), and TEF (OR419796), respectively. Phylogenetic analyses were performed by concatenating all the sequenced loci using the Bayesian method in Phylosuite (Zhang et al.2020). The phylogenetic tree indicated that the isolate belongs to the A. alternata clade with a bootstrap value of 75%. The pathogen was identified as A. alternata based on the morphological and molecular results. To satisfy Koch's postulates, a conidial suspension (106 conidia/mL) of the HY2-1 was prepared with ddH2O to infect the healthy plants. Ninety healthy leaves on 30 plants in pots were punctured using a sterilized needle, and then inoculated by spraying the conidial suspension on the wounded leaves in a greenhouse at 25°C and 80% relative humidity. The control plants were sprayed with ddH2O. The plants showed similar symptoms to the original infected plant 15 d after inoculation. The controls showed no symptoms. A pure culture of A. alternata was isolated and identified again as previously described. Leaf blight caused by A. alternata has been reported on Taro (Liu et al. 2020), Toona ciliata (Wang et al. 2023), etc. To our knowledge, this is the first report of E. sagittatum leaf blight caused by A. alternata in China. The results will help to develop effective control strategies for leaf blight on E. sagittatum.

Target isolation of diverse sesquiterpenoid from the stems of Daphne genkwa based on molecular networking.

Guiding by LC-MS/MS analysis and the Global Natural Products Social (GNPS) Molecular Networking, three undescribed sesquiterpenoids, stedapgens A-C, and two known analogues were discovered in the barks of Daphne genkwa Sieb. et Zucc. The structures were determined by analysis of their spectroscopic data and quantum-chemical calculations. All the isolated novel compounds were tested for their acetylcholinesterase inhibitory activities with IC50 = 0.754 ± 0.059, 0.696 ± 0.026, and 0.337 ± 0.023 µg/ml. Among them, stedapgen A displayed promising inhibitory activities against AChE, and the binding sites were predicted by molecular docking.

Anti-Inflammatory Effect of Chamaecyparis obtusa (Siebold & Zucc.) Endl. Leaf Essential Oil.

Chamaecyparis obtusa (Siebold & Zucc.) Endl. (C. obtusa) belongs to the Cupressaceae family and is native to East Asian regions. Essential oils extracted from the leaves, bark, branches, and roots of C. obtusa have both aesthetic and medicinal properties and are thus widely used. However, detailed analyses of the active ingredients of C. obtusa extract are lacking. In this study, the sabinene content in the hydro-distillation of C. obtusa leaf essential oil (COD) was analyzed using GC-MS, and the anti-inflammatory effect of COD was compared with that of pure sabinene. Cell viability was evaluated by MTT assay, and nitric oxide (NO) production was measured using Griess reagent. Relative mRNA and protein levels were analyzed using RT-qPCR and western blot, and secreted cytokines were analyzed using a cytokine array kit. The results showed that both COD and sabinene inhibited the expression of inducible nitric oxide synthase (iNOS) and the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 in lipopolysaccharide (LPS)-induced RAW 264.7 cells. COD and sabinene also reduced the production of pro-inflammatory cytokines interleukin (IL)-1ß, IL-6, IL-27, IL-1 receptor antagonist (IL-1ra), and granulocyte-macrophage colony-stimulating factor (GM-CSF). The anti-inflammatory mechanisms of COD and sabinene partially overlap, as COD was shown to inhibit MAPKs and the JAK/STAT axis, and sabinene inhibited MAPKs, thereby preventing LPS-induced macrophage activation.

Effects of bleeding of Actinidia arguta (Sieb. & Zucc) Planch. ex miq. on its plant growth, physiological characteristics and fruit quality.

Bleeding is as particularly a serious phenomenon in Actinidia arguta and has important effects on this plant's growth and development. Here we used A. arguta to study the effects of bleeding on the growth and development of leaves and fruits after a bleeding episode. We detect and analyze physiological indices of leaves and fruit after bleeding. The result revealed that the relative electrical conductivity and malondialdehyde (MDA) of leaves increased in treatment. Nitro blue tetrazolium chloride (NBT) and 3,3-diaminobenzidine (DAB) staining revealed the accumulation of reactive oxygen species (ROS) in leaves after bleeding. The chlorophyll content and photosynthetic parameter of plants were also decreased. In fruits, pulp and seed water content decreased after the damage, as did fruit vitamin C (Vc), soluble sugar content, and soluble solids content (SSC); the titratable acid content did not change significantly. We therefore conclude that bleeding affects the physiological indices of A. arguta. Our study provides a theoretical basis for understanding the physiological changes of A. arguta after bleeding episodes and laying a timely foundation for advancing research on A. arguta bleeding and long-term field studies should be executed in order to gain insights into underlying mechanisms.

Current and potential trends in the bioactive properties and health benefits of Prunus mume Sieb. Et Zucc: a comprehensive review for value maximization.

Prunus mume Sieb. Et Zucc (P. mume) is an acidic fruit native to China (named Chinese Mei or greengage plum). It is currently cultivated in several Asian countries, including Japan ("Ume"), Korea (Maesil), and Vietnam (Mai or Mo). Due to its myriad nutritional and functional properties, it is accepted in different countries, and its characteristics account for its commercialization. In this review, we summarize the information on the bioactive compounds from the fruit of P. mume and their structure-activity relationships (SAR); the pulp has the highest enrichment of bioactive chemicals. The nutritional properties of P. mume and the numerous uses of its by-products make it a potential functional food. P. mume extracts exhibit antioxidant, anticancer, antimicrobial, and anti-hyperuricaemic properties, cardiovascular protective effects, and hormone regulatory properties in various in vitro and in vivo assays. SAR shows that the water solubility, molecular weight, and chemical conformation of P. mume extracts are closely related to their biological activity. However, further studies are needed to evaluate the fruit's potential nutritional and functional therapeutic mechanisms. The industrial process of large-scale production of P. mume and its extracts as functional foods or nutraceuticals needs to be further optimized.

Multi-target bioactive compound screening from the infructescence of Platycarya strobilacea Sieb. et Zucc. by affinity chromatography using immobilized ß2 -adrenoceptor and muscarinic-3 acetylcholine receptor as the stationary phase.

As a main source for the recognition and identification of lead compounds, traditional Chinese medicine plays a pivotal role in preventing diseases for years. However, screening bioactive compounds from traditional Chinese medicine remains challenging because of the complexity of the systems and the occurrence of the synergic effect of the compounds. The infructescence of Platycarya strobilacea Sieb. et Zucc is prescribed for allergic rhinitis treatment with unknown bioactive compounds and unclear mechanisms. Herein, we immobilized the ß2 -adrenoceptor and muscarine-3 acetylcholine receptor onto the silica gel surface to prepare the stationary phase in a covalent bond through one step. The feasibility of the columns was investigated by the chromatographic method. Ellagic acid and catechin were identified as the bioactive compounds targeting the receptors. The binding constants of ellagic acid were calculated to be (1.56 ± 0.23)×107  M-1 for muscarine-3 acetylcholine receptor and (2.93 ± 0.15)×107  M-1 for ß2 -adrenoceptor by frontal analysis. While catechin can bind with muscarine-3 acetylcholine receptor with an affinity of (3.21 ± 0.05)×105  M-1 . Hydrogen bonds and van der Waals' force were the main driving forces for the two compounds with the receptors. The established method provides an alternative for multi-target bioactive compound screening in complex matrices.

Isolation of Pro-Osteogenic Compounds from Euptelea polyandra That Reciprocally Regulate Osteoblast and Osteoclast Differentiation.

Plants contain a large number of small-molecule compounds that are useful for targeting human health and in drug discovery. Healthy bone metabolism depends on the balance between bone-forming osteoblast activity and bone-resorbing osteoclast activity. In an ongoing study searching for 22 plant extracts effective against osteoporosis, we found that the crude extract of Euptelea polyandra Sieb. et Zucc (E. polyandra) had osteogenic bioactivity. In this study, we isolated two compounds, isoquercitrin (1) and astragalin (2), responsible for osteogenic bioactivity in osteoblastic MC3T3-E1 cells from the leaf of E. polyandra using column chromatography and the spectroscopic technique. This is the first report to isolate astragalin from E. polyandra. Compounds (1) and (2) promoted osteoblast differentiation by increasing alkaline phosphatase (ALP) activity and alizarin red S stain-positive calcium deposition, while simultaneously suppressing tartrate-resistant acid phosphatase (TRAP)-positive osteoclast differentiation in RAW264.7 cells at non-cytotoxic concentrations. Isoquercitrin (1) and astragalin (2) increased the expression of osteoblastic differentiation genes, Osterix, ALP, and Osteoprotegerin in the MC3T3-E1 cells, while suppressing osteoclast differentiation genes, TRAP, Cathepsin K, and MMP 9 in the RAW264.7 cells. These compounds may be ideal targets for the treatment of osteoporosis due to their dual function of promoting bone formation and inhibiting bone resorption.

New Type of Tannins Identified from the Seeds of Cornus officinalis Sieb. et Zucc. by HPLC-ESI-MS/MS.

There is a lack of information on the compound profile of Cornus officinalis Sieb. et Zucc. seeds. This greatly affects their optimal utilization. In our preliminary study, we found that the extract of the seeds displayed a strong positive reaction to the FeCl3 solution, indicating the presence of polyphenols. However, to date, only nine polyphenols have been isolated. In this study, HPLC-ESI-MS/MS was employed to fully reveal the polyphenol profile of the seed extracts. A total of 90 polyphenols were identified. They were classified into nine brevifolincarboxyl tannins and their derivatives, 34 ellagitannins, 21 gallotannins, and 26 phenolic acids and their derivatives. Most of these were first identified from the seeds of C. officinalis. More importantly, five new types of tannins were reported for the first time: brevifolincarboxyl-trigalloyl-hexoside, digalloyl-dehydrohexahydroxydiphenoyl (DHHDP)-hexdside, galloyl-DHHDP-hexoside, DHHDP-hexahydroxydiphenoyl(HHDP)-galloyl-gluconic acid, and peroxide product of DHHDP-trigalloylhexoside. Moreover, the total phenolic content was as high as 79,157 ± 563 mg gallic acid equivalent per 100 g in the seeds extract. The results of this study not only enrich the structure database of tannins, but also provide invaluable aid to its further utilization in industries.

Screening the Extract of Laportea bulbifera (Sieb. et Zucc.) Wedd. Based on Active Component Content, Its Antioxidant Capacity and Exploration of Hepatoprotective Activity in Rats.

Laportea bulbifera (Sieb. et Zucc.) Wedd., a plant with a long history of medicinal use, possesses uncertainly defined medicament portions while its antioxidant capacity remains largely unexplored. To gain a better understanding of its medicinal value, this study focused on investigating the Laportea bulbifera aboveground part (LBAP) and the Laportea bulbifera root (LBR). Through an assessment of the bioactive compound content, a significant finding emerged: the LBR exhibited notably higher levels of these bioactive phytochemicals compared to the LBAP. This observation was further reinforced by the antioxidant assays, which demonstrated the superiority of the LBR's antioxidant capacity. The experimental results unequivocally indicate that the root is the optimal medicament portion for Laportea bulbifera. Furthermore, it was discovered that the presence of alcohol in the extraction solvent significantly enhanced the extraction of active ingredients, with the methanol extract of LBR performing the best among the extracts tested. Consequently, this extract was selected for further research. Leveraging cutting-edge UHPLC-ESI-Q-TOF-MS technology, the methanol extract of LBR was meticulously analyzed, revealing the presence of 41 compounds, primarily belonging to the phenolics and fatty acids. Remarkably, stability experiments demonstrated that the phenolics in the methanol extract maintained their stability across various pH values and during in vitro simulations of the human digestive system, albeit showing gradual degradation under high temperatures. Furthermore, the oxidative stability tests conducted on oils revealed the potential of the methanol extract as a stabilizer for olive oil and sunflower oil. Moreover, oral acute toxicity studies confirmed the low toxicity of the methanol extract, further supporting its safe use for medicinal purposes. Of particular note, histopathological examination and biochemical analysis affirmed the remarkable protective effects of the methanol extract against d-galactosamine-induced liver damage. These findings underscore the therapeutic potential of the methanol extract from the LBR in the treatment of diseases associated with oxidative imbalance.

Actinidia arguta (Sieb. et Zucc.) Planch. ex Miq.: A Review of Phytochemistry and Pharmacology.

Actinidia arguta (Siebold & Zucc.) Planch ex Miq. (A. arguta) is a highly valued vine plant belonging to the Actinidia lindl genus. It is extensively utilized for its edible and medicinal properties. The various parts of A. arguta serve diverse purposes. The fruit is rich in vitamins, amino acids, and vitamin C, making it a nutritious and flavorful raw material for producing jam, canned food, and wine. The flowers yield volatile oils suitable for essential oil extraction. The leaves contain phenolic compounds and can be used for tea production. Additionally, the roots, stems, and leaves of A. arguta possess significant medicinal value, as they contain a wide array of active ingredients that exert multiple pharmacological and therapeutic effects. These effects include quenching thirst, relieving heat, stopping bleeding, promoting blood circulation, reducing swelling, dispelling wind, and alleviating dampness. Comprehensive information on A. arguta was collected from scientific databases covering the period from 1970 to 2023. The databases used for this review included Web of Science, PubMed, ProQuest, and CNKI. The objective of this review was to provide a detailed explanation of A. arguta from multiple perspectives, such as phytochemistry and pharmacological effects. By doing so, it aimed to establish a solid foundation and propose new research ideas for further exploration of the plant's potential applications and industrial development. To date, a total of 539 compounds have been isolated and identified from A. arguta. These compounds include terpenoids, flavonoids, phenolics, phenylpropanoids, lignin, organic acids, volatile components, alkanes, coumarins, anthraquinones, alkaloids, polysaccharides, and inorganic elements. Flavonoids, phenolics, alkaloids, and polysaccharides are the key bioactive constituents of A. arguta. Moreover, phenolics and flavonoids in A. arguta exhibit remarkable antioxidant, anti-inflammatory, and anti-tumor properties. Additionally, they show promising potential in improving glucose metabolism, combating aging, reducing fatigue, and regulating the immune system. While some fundamental studies on A. arguta have been conducted, further research is necessary to enhance our understanding of its mechanism of action, quality evaluation, and compatibility mechanisms. A more comprehensive investigation is highly warranted to explore the mechanism of action and expand the range of drug resources associated with A. arguta. This will contribute to the current hot topics of anti-aging and anti-tumor drug research and development, thereby promoting its further development and utilization.

Ethnomedicinal Uses, Geographical Distribution, Botanical Description, Phytochemistry, Pharmacology, and Quality Control of Laportea bulbifera (Sieb. et Zucc.) Wedd.: A Review.

Laportea bulbifera (Sieb. et Zucc.) Wedd. (L. bulbifera) is a significant plant in the Laportea genus. Traditionally, it has been used in ethnomedicine for treating various conditions such as rheumatic arthralgia, fractures, falling injuries, nephritis dropsy, limb numbness, pruritus, fatigue-induced internal imbalances, and irregular menstruation. Modern pharmacological studies have confirmed its therapeutic potential, including anti-inflammatory, immunosuppressive, analgesic, and anti-rheumatoid arthritis properties. To gather comprehensive information on L. bulbifera, a thorough literature search was conducted using databases like Web of Science, PubMed, ProQuest, and CNKI. This review aims to provide a comprehensive understanding of L. bulbifera, covering various aspects such as ethnomedicinal uses, geographical distribution, botanical description, phytochemistry, pharmacology, and quality control. The goal is to establish a solid foundation and propose new research avenues for exploring and developing potential applications of L. bulbifera. So far, a total of one hundred and eighty-nine compounds have been isolated and identified from L. bulbifera, including flavonoids, phenolics, nitrogen compounds, steroids, terpenoids, coumarins, phenylpropanoids, fatty acids and their derivatives, and other compounds. Notably, flavonoids and fatty acids have demonstrated remarkable antioxidant and anti-inflammatory properties. Additionally, these compounds show promising potential in activities such as analgesia, hypoglycemia, and hypolipidemia, as well as toxicity. Despite extensive fundamental studies on L. bulbifera, further research is still needed to enhance our understanding of its mechanism of action and improve quality control. This requires more comprehensive investigations to explore the specific material basis, uncover new mechanisms of action, and refine quality control methods related to L. bulbifera. By doing so, we could contribute to the further development and utilization of this plant.

Impact of Fermentation, Autoclaving and Phytase Treatment on the Antioxidant Properties and Quality of Teff Cookies.

Research background: Teff [Eragrostis tef (Zucc.) Trotter] is an underutilised cereal crop grown mainly in Ethiopia and Eritrea. It is an excellent source of dietary fibre, vitamins, minerals and bioactive compounds. However, it also contains a high amount of phytic acid, which is an antinutrient and reduces the bioavailability of minerals and proteins. To improve the nutritional quality of teff, the phytic acid content should be reduced by an effective dephytinisation method. Experimental approach: In this study, various dephytinisation methods (fermentation, autoclaving and phytase treatment) were used to dephytinise teff flour. Undephytinised and dephytinised teff flour was mixed into wheat flour (0-40 %) to improve the functional properties of cookies. Twenty different cookie formulations were prepared according to 4x5x2 factorial design. The physical, chemical, nutritional and sensory properties of the cookies were investigated. Results and conclusions: Among the dephytinisation methods, fermentation produced the most effective reduction in phytic acid mass fraction (181 mg/100 g), followed by phytase treatment (198 mg/100 g). The protein, fat, Fe and Zn content and antioxidant activity of cookies enriched with dephytinised teff flour were comparable to cookies fortified with undephytinised teff flour. Moreover, the dephytinised teff cookies had lower phytic acid mass fractions. The cookies containing 40 % teff flour had higher antioxidant activity and nutritional quality than the control wheat cookies. The use of dephytinised teff flour reduced the spread ratio and the a* and b* values of cookies compared to undephytinised flour. Cookies containing fermented and phytase-treated teff flour had a harder texture than cookies containing undephytinised flour. In addition, as the amount of teff flour increased, the spread ratio values of cookies gradually incrased while their hardness decreased. Overall acceptability scores of cookies containing 10-20 % teff flour were similar to the control. Novelty and scientific contribution: To the best of our knowledge, this is the first study to determine the quality of cookies containing dephytinised teff flour. The data highlight the potential of dephytinised (especially autoclaved and phytase-treated) teff flour (up to 20 %) as a functional ingredient to enrich the mineral content and antioxidant capacity of foods. Furthermore, this study shows that fermentation, autoclaving and phytase treatment can be used to improve the nutritional quality of grains.

Characterization of the chemical constituents and metabolic profile of Polygonum cuspidatum Sieb. et Zucc. in rat plasma, urine, and feces by ultra-high performance liquid chromatography coupled with Quadrupole-Exactive Orbitrap mass spectrometry.

Polygonum cuspidatum Sieb. et Zucc. is a traditional and popular Chinese medicine with a wide spectrum of pharmacological effects such as anti-bacterial, anti-inflammatory, and anti-tumor activities together with other health effects like lowering lipids, preventing diabetes, and regulating the immune system. It is of great significance to explore the complex chemical constituents and metabolic process of Polygonum cuspidatum in vivo to further clarify the effective substances. However, studies on its metabolism in vivo were not comprehensive in previous literature. In this study, ultra-high performance liquid chromatography coupled with Quadrupole-Exactive Orbitrap mass spectrometry was used to comprehensively identify the chemical constituents in Polygonum cuspidatum and further analyze its metabolic profile in rats. Compared with reference substances, various databases, and literature retrieval, 62 compounds were inferred from the Polygonum cuspidatum extract. Furthermore, a total of 119 compounds, including 44 prototype compounds and 75 metabolites, were annotated in rat plasma, urine, and feces. The main metabolic pathways of Polygonum cuspidatum in rats included hydrogenation reduction, hydroxylation, dehydration, methylation, sulfation, and glucuronidation. This is the first systematic study on the chemical constituents of Polygonum cuspidatum and its metabolic profile in vivo, which contributes to finding its bioactive components and seeking its therapeutic targets.

Genome-Wide Association Studies Reveal Novel Loci for Herbivore Resistance in Wild Soybean (Glycine soja).

The production of soybean [Glycine max (L.) Merr.] is seriously threatened by various leaf-feeding insects, and wild soybean [Glycine soja Sieb. & Zucc.] has a greater resistance capacity and genetic diversity. In this study, a natural population consisting of 121 wild soybean accessions was used for detecting insect resistance genes. The larval weight (LW) of the common cutworm (CCW), the resistance level (RL) and the index of damaged leaf (IDL) were evaluated as resistance indicators to herbivores. An association synonymous SNP AX-94083016 located in the coding region of the respiratory burst oxidase gene GsRbohA1 was identified by genome-wide association study (GWAS) analyses. The overexpression of GsRbohA1 in soybean hairy roots enhanced resistance to CCW. One SNP in the promoter region cosegregated with AX-94083016 contributing to soybean resistance to CCW by altering GsRbohA1 gene expression and reactive oxygen species (ROS) accumulation. Two major haplotypes, GsRbohA1A and GsRbohA1G, were identified based on the SNP. The resistant haplotype GsRbohA1A predominates in wild soybeans, although it has been gradually lost in landraces and cultivars. The nucleotide diversity around GsRbohA1 is much lower in landraces and cultivars than in its ancestors. In conclusion, a new resistant haplotype, GsRbohA1A, was identified in wild soybean, which will be a valuable gene resource for soybean insect resistance breeding through introducing into improvement lines, and it offers a strategy for exploring resistance gene resources from its wild relatives.

Acteoside and ursolic acid synergistically protects H2O2-induced neurotrosis by regulation of AKT/mTOR signalling: from network pharmacology to experimental validation.

CONTEXT: Ursolic acid (UA) and acteoside (ATS) are important active components that have been used to treat Alzheimer's disease (AD) because of their neuroprotective effects, but the exact mechanism is still unclear. OBJECTIVE: Network pharmacology was used to explore the mechanism of UA + ATS in treating AD, and cell experiments were used to verify the mechanism. MATERIALS AND METHODS: UA + ATS targets and AD-related genes were retrieved from TCMSP, STITCH, SwissTargetPrediction, GeneCards, DisGeNET and GEO. Key targets were obtained by constructing protein interaction network through STRING. The neuroprotective effects of UA + ATS were verified in H2O2-treated PC12 cells. The subsequent experiments were divided into Normal, Model (H2O2 pre-treatment for 4 h), Control (H2O2+ solvent pre-treatment), UA (5 µM), ATS (40 µM), UA (5 µM) + ATS (40 µM). Then apoptosis, mitochondrial membrane potential, caspase-3 activity, ATG5, Beclin-1 protein expression and Akt, mTOR phosphorylation levels were detected. RESULTS: The key targets of UA + ATS-AD network were mainly enriched in Akt/mTOR pathway. Cell experiments showed that UA (ED50: 5 µM) + ATS (ED50: 40 µM) could protect H2O2-induced (IC50: 250 µM) nerve damage by enhancing cells viability, combating apoptosis, restoring MMP, reducing the activation of caspase-3, lessening the phosphorylation of Akt and mTOR, and increasing the expression of ATG5 and Beclin-1. CONCLUSIONS: ATS and UA regulates multiple targets, bioprocesses and signal pathways against AD pathogenesis. ATS and UA synergistically protects H2O2-induced neurotrosis by regulation of AKT/mTOR signalling.

Transcriptome Analysis of Salt Stress in Hibiscus hamabo Sieb. et Zucc Based on Pacbio Full-Length Transcriptome Sequencing.

Hibiscus hamabo Sieb. et Zucc is an important semi-mangrove plant with great morphological features and strong salt resistance. In this study, by combining single molecule real time and next-generation sequencing technologies, we explored the transcriptomic changes in the roots of salt stressed H. hamabo. A total of 94,562 unigenes were obtained by clustering the same isoforms using the PacBio RSII platform, and 2269 differentially expressed genes were obtained under salt stress using the Illumina platform. There were 519 differentially expressed genes co-expressed at each treatment time point under salt stress, and these genes were found to be enriched in ion signal transduction and plant hormone signal transduction. We used Arabidopsis thaliana (L.) Heynh. transformation to confirm the function of the HhWRKY79 gene and discovered that overexpression enhanced salt tolerance. The full-length transcripts generated in this study provide a full characterization of the transcriptome of H. hamabo and may be useful in mining new salt stress-related genes specific to this species, while facilitating the understanding of the salt tolerance mechanisms.

Identifying Wild Versus Cultivated Gene-Alleles Conferring Seed Coat Color and Days to Flowering in Soybean.

Annual wild soybean (G. soja) is the ancestor of the cultivated soybean (G. max). To reveal the genetic changes from soja to max, an improved wild soybean chromosome segment substitution line (CSSL) population, SojaCSSLP5, composed of 177 CSSLs with 182 SSR markers (SSR-map), was developed based on SojaCSSLP1 generated from NN1138-2(max)×N24852(soja). The SojaCSSLP5 was genotyped further through whole-genome resequencing, resulting in a physical map with 1366 SNPLDBs (SNP linkage-disequilibrium blocks), which are composed of more markers/segments, shorter marker length and more recombination breakpoints than the SSR-map and caused 721 new wild substituted segments. Using the SNPLDB-map, two loci co-segregating with seed-coat color (SCC) and six loci for days to flowering (DTF) with 88.02% phenotypic contribution were identified. Integrated with parental RNA-seq and DNA-resequencing, two SCC and six DTF candidate genes, including three previously cloned (G, E2 and GmPRR3B) and five newly detected ones, were predicted and verified at nucleotide mutant level, and then demonstrated with the consistency between gene-alleles and their phenotypes in SojaCSSLP5. In total, six of the eight genes were identified with the parental allele-pairs coincided to those in 303 germplasm accessions, then were further demonstrated by the consistency between gene-alleles and germplasm phenotypes. Accordingly, the CSSL population integrated with parental DNA and RNA sequencing data was demonstrated to be an efficient platform in identifying candidate wild vs. cultivated gene-alleles.

Anti-Gastritis and Anti-Lung Injury Effects of Pine Tree Ethanol Extract Targeting Both NF-κB and AP-1 Pathways.

An ethanol extract (Pd-EE) of Pinus densiflora Siebold and Zucc was derived from the branches of pine trees. According to the Donguibogam, pine resin has the effects of lowering the fever, reducing pain, and killing worms. The purpose of this study is to investigate whether Pd-EE has anti-inflammatory effects. During in vitro trials, NO production, as well as changes in the mRNA levels of inflammation-related genes and the phosphorylation levels of related proteins, were confirmed in RAW264.7 cells activated with lipopolysaccharide depending on the presence or absence of Pd-EE treatment. The activities of transcription factors were checked in HEK293T cells transfected with adapter molecules in the inflammatory pathway. The anti-inflammatory efficacy of Pd-EE was also estimated in vivo with acute gastritis and acute lung injury models. LC-MS analysis was conducted to identify the components of Pd-EE. This extract reduced the production of NO and the mRNA expression levels of iNOS, COX-2, and IL-6 in RAW264.7 cells. In addition, protein expression levels of p50 and p65 and phosphorylation levels of FRA1 were decreased. In the luciferase assay, the activities of NF-κB and AP-1 were lowered. In acute gastritis and acute lung injury models, Pd-EE suppressed inflammation, resulting in alleviated damage.

Gastrodia elata Blume and Zanthoxylum schinifolium Siebold & Zucc Mixed Extract Suppress Platelet Aggregation and Thrombosis.

Background and objectives: Blood vessel thrombosis causes blood circulation disorders, leading to various diseases. Currently, various antiplatelet and anticoagulant drugs, such as aspirin, warfarin, heparin, and non-vitamin K antagonist oral anticoagulants (NOACs), are used as the major drugs for the treatment of a wide range of thrombosis. However, these drugs have a side effect of possibly causing internal bleeding due to poor hemostasis when taken for a long period of time. Materials and Methods: Gastrodia elata Blume (GE) and Zanthoxylum schinifolium Siebold & Zucc (ZS) are known to exhibit hemostatic and antiplatelet effects as traditional medicines that have been used for a long time. In this study, we investigated the effect of a mixed extract of GE and ZS (MJGE09) on platelet aggregation and plasma coagulation. Results: We found that MJGE09 inhibited collagen-and ADP-induced platelet aggregation in vitro. In addition, collagen- and ADP-induced platelet aggregation were also inhibited in a dose-dependent manner on the platelets of mice that were orally administered MJGE09 ex vivo. However, compared with aspirin, MJGE09 did not prolong the rat tail vein bleeding time in vivo and did not show a significant effect on the increase in the prothrombin time (PT) and activated partial thromboplastin time (aPTT). Conclusions: These results suggest that MJGE09 can be used as a potential anticoagulant with improved antithrombotic efficacy.

Anti-Proliferative Effects of Standardized Cornus officinalis on Benign Prostatic Epithelial Cells via the PCNA/E2F1-Dependent Cell Cycle Pathway.

Cornus officinalis, widely used in traditional Chinese medicine, exhibits pharmacological effects against erectile dysfunction and pollakisuria, which are pathological symptoms of benign prostatic hyperplasia (BPH). Although traditional usage and a study on BPH have been reported, to our knowledge, no study has investigated the exact molecular mechanism(s) underlying the anti-proliferative effects of standardized C. officinalis on prostatic cells. We standardized C. officinalis 30% ethanol extract (COFE) and demonstrated the therapeutic effects of COFE on human BPH epithelial cells and testosterone-induced BPH in rats. In vitro studies using BPH-1 cells demonstrated an upregulation of BPH-related and E2F Transcription Factor 1(E2F1)-dependent cell cycle markers, whereas treatment with COFE clearly inhibited the proliferation of BPH epithelial cells and reduced the overexpression of G1 and S checkpoint genes. Additionally, COFE administration alleviated the androgen-dependent prostatic enlargement in a testosterone-induced BPH animal model. COFE exerted these anti-BPH effects by the inhibition of anti-apoptotic markers, suppression of PCNA expression, and regulation of E2F1/pRB-dependent cell cycle markers in rats with BPH. These results suggest that COFE exerts anti-proliferative effect by regulating PCNA/E2F1-dependent cell cycle signaling pathway both in vivo and in vitro. These findings reveal the therapeutic potential of COFE, which could be used as a substitute for BPH treatment.