Pharmacological and Pathological Effects of Mulberry Leaf Extract on the Treatment of Type 1 Diabetes Mellitus Mice.

This study investigated the pharmacological and pathological effects of aqueous mulberry leaf extract on type 1 diabetes mellitus mice induced with an intraperitoneal injection of streptozotocin (STZ). Diabetic mice were randomized into six groups: control (normal group), model, metformin-treated mice, and high-dose, medium-dose, and low-dose mulberry. The mulberry-treated mice were divided into high-, medium-, and low-dose groups based on the various doses of aqueous mulberry leaf extract during gavage. The efficacy of the six-week intervention was evaluated by measuring levels of fasting plasma glucose, alkaline phosphatase, alanine aminotransferase, aspartate transaminase, blood urea nitrogen, gamma-glutamyl transferase, glucose, high-density lipoprotein cholesterol, lactate dehydrogenase, and low-density lipoprotein cholesterol and recording body weight. Results revealed that mulberry leaf extract exhibited an ideal hypoglycemic effect, and the high-dose group was the most affected. Histology analysis, glycogen staining and apoptosis detection were used to study the extract's effects on the liver, kidney, and pancreatic cells of diabetic mice, enabling the assessment of its effectiveness and complications on a clinical and theoretical basis. It was shown that a certain concentration of aqueous mulberry leaf extract repaired the islet cells of type 1 diabetes mellitus mice, promoting normal insulin secretion. Herein, it was confirmed that mulberry leaf could be used to develop new hypoglycemic drugs or functional health food with broad applicability.

Study on Process Optimization and Antioxidant Activity of Polysaccharide from Bletilla striata Extracted via Deep Eutectic Solvents.

Taking the extraction yield of Bletilla striata polysaccharide (BSP) as the index and taking the type of deep eutectic solvents (DESs), extraction time, extraction temperature, DES water content, and solid-liquid ratio as the investigation factors, single-factor and Box-Behnken response surface tests were carried out to optimize the extraction process of BSP. Thus, the antioxidant activity of BSP on DPPH radicals, ABTS radicals and ferric reducing antioxidant power were determined. The results showed that the most suitable deep eutectic solvent was DES-2, namely choline chloride-urea. The optimal extraction conditions for BSP were an extraction time of 47 min, extraction temperature of 78 °C, water content of 35%, and solid-liquid ratio of 1:25. Under this optimized condition, the extraction yield of BSP was able to reach (558.90 ± 8.83) mg/g, and recycling studies indicated the good cycle stability of the DES. Antioxidant results showed that BSP had superior antioxidant activity and had a dose-response relationship with drug concentration. Compared with Bletilla striata polysaccharide obtained via conventional hot water extraction (BSP-W), the extraction yield of BSP obtained through this method (BSP-2) increased by 36.77%, the scavenging activity of DPPH radicals increased by 24.99%, the scavenging activity of ABTS radicals increased by 41.16%, and the ferric reducing antioxidant power increased by 49.19%. Therefore, DESs as new green reagents and BSP extracted with DESs not only had a high yield but also had strong antioxidant activity.

Effective identification of varieties by nucleotide polymorphisms and its application for essentially derived variety identification in rice.

BACKGROUND: Plant variety identification is the one most important of agricultural systems. Development of DNA marker profiles of released varieties to compare with candidate variety or future variety is required. However, strictly speaking, scientists did not use most existing variety identification techniques for "identification" but for "distinction of a limited number of cultivars," of which generalization ability always not be well estimated. Because many varieties have similar genetic backgrounds, even some essentially derived varieties (EDVs) are involved, which brings difficulties for identification and breeding progress. A fast, accurate variety identification method, which also has good performance on EDV determination, needs to be developed. RESULTS: In this study, with the strategy of "Divide and Conquer," a variety identification method Conditional Random Selection (CRS) method based on SNP of the whole genome of 3024 rice varieties was developed and be applied in essentially derived variety (EDV) identification of rice. CRS is a fast, efficient, and automated variety identification method. Meanwhile, in practical, with the optimal threshold of identity score searched in this study, the set of SNP (including 390 SNPs) showed optimal performance on EDV and non-EDV identification in two independent testing datasets. CONCLUSION: This approach first selected a minimal set of SNPs to discriminate non-EDVs in the 3000 Rice Genome Project, then united several simplified SNP sets to improve its generalization ability for EDV and non-EDV identification in testing datasets. The results suggested that the CRS method outperformed traditional feature selection methods. Furthermore, it provides a new way to screen out core SNP loci from the whole genome for DNA fingerprinting of crop varieties and be useful for crop breeding.

Bioinformatics-assisted, integrated omics studies on medicinal plants.

The immense therapeutic and economic values of medicinal plants have attracted increasing attention from the worldwide researchers. It has been recognized that production of the authentic and high-quality herbal drugs became the prerequisite for maintaining the healthy development of the traditional medicine industry. To this end, intensive research efforts have been devoted to the basic studies, in order to pave a way for standardized authentication of the plant materials, and bioengineering of the metabolic pathways in the medicinal plants. In this paper, the recent advances of omics studies on the medicinal plants were summarized from several aspects, including phenomics and taxonomics, genomics, transcriptomics, proteomics and metabolomics. We proposed a multi-omics data-based workflow for medicinal plant research. It was emphasized that integration of the omics data was important for plant authentication and mechanistic studies on plant metabolism. Additionally, the computational tools for proper storage, efficient processing and high-throughput analyses of the omics data have been introduced into the workflow. According to the workflow, authentication of the medicinal plant materials should not only be performed at the phenomics level but also be implemented by genomic and metabolomic marker-based examination. On the other hand, functional genomics studies, transcriptional regulatory networks and protein-protein interactions will contribute greatly for deciphering the secondary metabolic pathways. Finally, we hope that our work could inspire further efforts on the bioinformatics-assisted, integrated omics studies on the medicinal plants.

Organ-specific, integrated omics data-based study on the metabolic pathways of the medicinal plant Bletilla striata (Orchidaceae).

BACKGROUND: Bletilla striata is one of the important species belonging to the Bletilla genus of Orchidaceae. Since its extracts have an astringent effect on human tissues, B. striata is widely used for hemostasis and healing. Recently, some other beneficial effects have also been uncovered, such as antioxidation, antiinflammation, antifibrotic, and immunomodulatory activities. As a key step towards a thorough understanding on the medicinal ingredient production in B. striata, deciphering the regulatory codes of the metabolic pathways becomes a major task. RESULTS: In this study, three organs (roots, tubers and leaves) of B. striata were analyzed by integrating transcriptome sequencing and untargeted metabolic profiling data. Five different metabolic pathways, involved in polysaccharide, sterol, flavonoid, terpenoid and alkaloid biosynthesis, were investigated respectively. For each pathway, the expression patterns of the enzyme-coding genes and the accumulation levels of the metabolic intermediates were presented in an organ-specific way. Furthermore, the relationships between enzyme activities and the levels of the related metabolites were partially inferred. Within the biosynthetic pathways of polysaccharides and flavonoids, long-range phytochemical transportation was proposed for certain metabolic intermediates and/or the enzymes. CONCLUSIONS: The data presented by this work could strengthen the molecular basis for further studies on breeding and medicinal uses of B. striata.

Investigating microRNA-mediated regulation of the nascent nuclear transcripts in plants: a bioinformatics workflow.

Most of the microRNAs (miRNAs) play their regulatory roles through posttranscriptional target decay or translational inhibition. For both plants and animals, these regulatory events were previously considered to take place in cytoplasm, as mature miRNAs were observed to be exported to the cytoplasm for Argonaute (AGO) loading and subsequent target binding. Recently, this notion was challenged by increasing pieces of evidence in the animal cells that uncovered the nuclear importation and action of the AGO-associated miRNAs. The nuclear-localized regulatory mode was also reported for the plant miRNAs. However, evidence is still lacking to show the universality and conservation of the miRNA-mediated regulation in the plant nuclei. Here, we introduced a bioinformatics workflow for genome-wide investigation of miRNA-guided, cleavage-based regulation of the nascent nuclear transcripts. Facilitated by the tool package PmiRNTSA (Plant microRNA-mediated nascent transcript slicing analyzer), plant biologists could perform a comprehensive search for the miRNA slicing sites located within the introns or the exon-intron/intron-exon junctions of the target transcripts, which are supported by degradome sequencing data. The results enable the researchers to examine the co-transcriptional regulatory model of the miRNAs for a specific plant species. Moreover, a case study was performed to search for the slicing sites located within the exon-intron/intron-exon junctions in two model plants. A case study was performed to show the feasibility and reliability of our workflow. Together, we hope that this work could inspire much more innovative research efforts to expand the current understanding of the miRNA action modes in plants.

The RNA degradome: a precious resource for deciphering RNA processing and regulation codes in plants.

The plant RNA degradome was defined as an aggregate of the RNA fragments degraded from various biochemical pathways, such as RNA turnover, maturation and quality surveillance. In recent years, the degradome sequencing (degradome-seq) libraries became a rich storehouse for researchers to study on RNA processing and regulation. Here, we provided a brief overview of the uses of degradome-seq data in plant RNA biology, especially on non-coding RNA processing and small RNA-guided target cleavages. Some novel applications in RNA research area, such as in vivo mapping of the endoribonucleolytic cleavage sites, identification of conserved motifs at the 5' ends of the uncapped RNA fragments, and searching for the protein-binding regions on the transcripts, were also mentioned. More importantly, we proposed a model for the biologists to deduce the contributions of transcriptional and/or post-transcriptional regulation to gene differential expression based on degradome-seq data. Finally, we hope that the degradome-based analytical methods could be widely applied for the studies on RNA biology in eukaryotes.

New glycosides from the twigs and leaves of Rhododendron latoucheae.

Six new glycosides (1-6), together with three known ones, were isolated from the twigs and leaves of Rhododendron latoucheae. Their structures were elucidated based on the spectroscopic data, including infrared spectrometry, mass spectrometry, and nuclear magnetic resonance experiments, along with Mosher's method. In addition, all compounds were tested their antiviral (herpes simplex virus-1 and influenza A/95-359) activities.

Minor Nortriterpenoids from the Twigs and Leaves of Rhododendron latoucheae.

A hyphenated NMR technique (analytical HPLC with a DAD connected to MS, SPE, and NMR) has proven effective for the full structural analysis and identification of minor natural products in complex mixtures. Application of this hyphenated technique to the CH2Cl2-soluble fraction of Rhododendron latoucheae led to the identification of 15 new minor ursane-type 28-nortriterpenoids (1-15). Compounds 1 and 12 inhibited HSV-1 with IC50 values of 6.4 and 0.4 µM, respectively.

The Gut Bacterial Community Composition of Wild Cervus albirostris (White-Lipped Deer) Detected by the 16S Ribosomal RNA Gene Sequencing.

Cervus albirostris (white-lipped deer) is an endemic species in China. As the name implies, C. albirostris has a characteristic pure white marking around their mouth and on the underside of the throat. The animal is a typical alpine species normally living at the height of 3500-4300 m. In this study, by pyrosequencing the 16S rRNA gene sequences, we for the first time analyzed the gut bacterial community composition in eight feces samples of wild C. albirostris. From a total of 243,634 high-quality sequences, we identified 186 genera, included in 17 prokaryotic phyla in the feces. The relative proportions of Firmicutes and Bacteroidetes were highly consistent in each individual sample. The most frequently detected genus was Ruminococcaceae UCG-005, ranging from 6.70 to 21.00%, displaying positively connections with the Rikenellaceae RC9 gut group. The bacterial communities associated with C. albirostris provide the basic knowledge for further microbiological studies and facilitates the conservation efforts of this vulnerable deer species.

Intronic regions of plant genes potentially encode RDR (RNA-dependent RNA polymerase)-dependent small RNAs.

Recent research has linked the non-coding intronic regions of plant genes to the production of small RNAs (sRNAs). Certain introns, called 'mirtrons' and 'sirtrons', could serve as the single-stranded RNA precursors for the generation of microRNA and small interfering RNA, respectively. However, whether the intronic regions could serve as the template for double-stranded RNA synthesis and then for sRNA biogenesis through an RDR (RNA-dependent RNA polymerase)-dependent pathway remains unclear. In this study, a genome-wide search was made for the RDR-dependent sRNA loci within the intronic regions of the Arabidopsis genes. Hundreds of intronic regions encoding three or more RDR-dependent sRNAs were found to be covered by dsRNA-seq (double-stranded RNA sequencing) reads, indicating that the intron-derived sRNAs were indeed generated from long double-stranded RNA precursors. More interestingly, phase-distributed sRNAs were discovered on some of the dsRNA-seq read-covered intronic regions, and those sRNAs were largely 24 nt in length. Based on these results, the opinion is put forward that the intronic regions might serve as the genomic origins for the RDR-dependent sRNAs. This opinion might add a novel layer to the current biogenesis model of the intron-derived sRNAs.

The use of high-throughput sequencing methods for plant microRNA research.

MicroRNA (miRNA) acts as a critical regulator of gene expression at post-transcriptional and occasionally transcriptional levels in plants. Identification of reliable miRNA genes, monitoring the procedures of transcription, processing and maturation of the miRNAs, quantification of the accumulation levels of the miRNAs in specific biological samples, and validation of miRNA-target interactions become the basis for thoroughly understanding of the miRNA-mediated regulatory networks and the underlying mechanisms. Great progresses have been achieved for sequencing technology. Based on the high degree of sequencing depth and coverage, the high-throughput sequencing (HTS, also called next-generation sequencing) technology provides unprecedentedly efficient way for genome-wide or transcriptome-wide studies. In this review, we will introduce several HTS platform-based methods useful for plant miRNA research, including RNA-seq (RNA sequencing), RNA-PET-seq (paired end tag sequencing of RNAs), sRNA-seq (small RNA sequencing), dsRNA-seq (double-stranded RNA sequencing), ssRNA-seq (single-stranded RNA sequencing) and degradome-seq (degradome sequencing). In particular, we will provide some special cases to illustrate the novel use of HTS methods for investigation of the processing modes of the miRNA precursors, identification of the RNA editing sites on miRNA precursors, mature miRNAs and target transcripts, re-examination of the current miRNA registries, and discovery of novel miRNA species and novel miRNA-target interactions. Summarily, we opinioned that integrative use of the above mentioned HTS methods could make the studies on miRNAs more efficient.

Deep-Eutectic-Solvent-in-Water Pickering Emulsions Stabilized by Starch Nanoparticles.

Deep eutectic solvents (DESs) have received extensive attention in green chemistry because of their ease of preparation, cost-effectiveness, and low toxicity. Pickering emulsions offer advantages such as long-term stability, low toxicity, and environmental friendliness. The oil phase in some Pickering emulsions is composed of solvents, and DESs can serve as a more effective alternative to these solvents. The combination of DESs and Pickering emulsions can improve the applications of green chemistry by reducing the use of harmful chemicals and enhancing sustainability. In this study, a Pickering emulsion consisting of a DES (menthol:octanoic acid = 1:1) in water was prepared and stabilized using starch nanoparticles (SNPs). The emulsion was thoroughly characterized using various techniques, including optical microscopy, transmission microscopy, laser particle size analysis, and rheological measurements. The results demonstrated that the DES-in-water Pickering emulsion stabilized by the SNPs had excellent stability and retained its structural integrity for more than 200 days at room temperature (20 °C). This prolonged stability has significant implications for many applications, particularly in the field of storage and transportation. This Pickering emulsion based on DESs and SNPs is sustainable and stable, and it has great potential to improve green chemistry practices in various fields.

Effect of stigmasterol and polyglycerol polyricinoleate concentrations on the preparation and properties of rapeseed oil-based gel emulsions.

Emulsion gels mimic the rheological properties of solid and semi-solid fats, offering a viable solution to replace conventional fats in low-fat food formulations. In this study, gel emulsions stabilized with stigmasterol (ST) and polyglycerol polyricinoleate (PGPR) complexes were prepared. Initially, we examined the effect of the ST/PGPR complex on the mechanism of gel emulsion stabilization. Our findings revealed that the gel emulsion formulated with 3% PGPR and ST exhibited a robust structure, effectively stabilizing the entire system and ensuring uniform distribution, and increasing ST concentration led to greater stability of the gel emulsion system. Stability assessments demonstrated that gel emulsions containing 3% PGPR and varying ST concentrations exhibited remarkable thermal stability and effectively delayed oil oxidation. These results underscore the high stability of gel emulsions stabilized with the ST/PGPR complex, highlighting their potential as a margarine substitute.

The influence of residual pectin composition and content on nanocellulose films from ramie fibers: Micro-nano structure and physical properties.

In this study, cellulose nanofibril (CNF) films from ramie fibers were prepared with different pectin compositions and contents, and the influence of residual pectin on the overall performances of CNF films was evaluated. There was no significant effect of the residual pectin composition on the properties of obtained CNF films. However, when the content of residual pectin was increased from 0.45 % to 9.16 %, the surface area and water absorption of CNF films were increased from 0.2223 to 0.3300 m2/g, and from 93.51 % to 122.42 %, respectively. Pectin covers the CNF surface and act as a physical barrier between the cellulose fibrils; thus the nanocellulose films with high pectin content will have a loose and porous structure, resulting in a high surface area and a high water absorption. Besides, with the residual pectin content decreasing from 9.16 % to 0.45 %, the UVA light transmittance and tensile strength of CNF films were increased from 30.6 % to 59.9 %, and from 37.67 to 100.26 MPa, respectively. After removal of amorphous pectins in CNFs, the low pectin containing CNFs are able to pack more compactly to form a strong and thin film. This paper provides guidance for the preparation of CNF films with different performance requirements.

An organ-specific transcriptomic atlas of the medicinal plant Bletilla striata: Protein-coding genes, microRNAs, and regulatory networks.

As one of the important species belonging to the Bletilla genus of Orchidaceae, Bletilla striata (Thunb.) Rchb. f., possesses both ornamental and medicinal values. Its dried tubers are used as a traditional Chinese medicine, and several secondary metabolites have been indicated to be the active ingredients. However, the molecular mechanisms related to the regulation of secondary metabolism have not been characterized in B. striata. In this study, integrated analysis of RNA sequencing (RNA-seq), small RNA sequencing (sRNA-seq), and degradome sequencing (degradome-seq) data from three organs (leaf, root, and tuber) of B. striata provided us with a comprehensive view of the microRNA (miRNA)-mediated regulatory network. Firstly, based on the RNA-seq data, the organ-specific expression patterns of the protein-coding genes, especially for those related to secondary metabolism, were investigated. Secondly, 342 conserved miRNA candidates were identified from B. striata. These miRNAs were assigned to 88 families, some of which were selected for expression pattern analysis. Additionally, 31 hairpin-structured precursors encoding 23 novel miRNAs were uncovered from the transcriptome assembly. Thirdly, based on the degradome signatures, 1,142 validated miRNA-target pairs (involving 167 conserved miRNAs and six novel miRNAs and 51 target genes) were included in the regulatory network. Organ-specific expression level comparison between the miRNAs and their targets revealed some interesting miRNA-target pairs. Fourthly, some valuable subnetworks were extracted for further functional studies. Additionally, some regulatory pathways were indicated to be monocot specific. Summarily, our results lay a solid basis for in-depth studies on the regulatory mechanisms underlying the production of the medicinal ingredients in B. striata.

RNA-, sRNA-, and degradome-seq were performed for three organs of B. striata. Organ-specific expression patterns of the protein-coding genes were analyzed. A total of 365 miRNAs were identified and subject to expression pattern analysis. A total of 1,142 miRNA-target pairs were validated for network construction. Some miRNA-mediated regulatory pathways were indicated to be monocot specific.

Bioactive Components From Gracilaria rubra With Growth Inhibition on HCT116 Colon Cancer Cells and Anti-inflammatory Capacity in RAW 264.7 Macrophages.

Gracilaria rubra is rich in bioactive compounds with various potential health benefits. This study aimed to elucidate the profile of both extractable bioactive components (EBCs) and non-extractable bioactive components (NEBCs) of G. rubra and determine their anti-colon cancer and anti-inflammatory activities. Both EBCs and NEBCs displayed strong suppressive effects on the viability of HCT116 cells, which causes cell cycle arrest, induces cellular apoptosis, and regulates the expression of cyclin-dependent kinases (CDKs) and tumor suppressor proteins. Additionally, EBCs and NEBCs from G. rubra displayed anti-inflammatory functions via inhibiting the production of nitric oxide (NO), reactive oxygen species (ROS), and proinflammatory cytokines in activated macrophages and regulating the expression levels of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), NADPH-quinone oxidoreductase-1 (NQO-1), and heme oxygenase 1 (HO-1). These findings provide a rationale for animal and human studies designed to evaluate the chemopreventive and anti-inflammatory potential of these bioactive compounds from G. rubra.

Inhibitory Effects of Polyphenols-Rich Components From Three Edible Seaweeds on Inflammation and Colon Cancer in vitro.

Polyphenols from edible seaweeds display various health benefits which have not been adequately studied. This study aimed to characterize the composition of extractable polyphenol-rich components (EPCs) and non-extractable polyphenol-rich components (NEPCs) from three edible seaweeds (i.e., Laminaria japonica, Ulva lactuca, and Porphyra tenera) and evaluate their anti-inflammatory capacities in activated macrophages and anticancer properties in colon cancer cells. Both EPCs and NEPCs from three edible seaweeds against lipopolysaccharides (LPS) stimulated nitric oxide in activated macrophages. Immunoblotting and qRT-PCR indicated that EPCs and NEPCs regulated the expression levels of proinflammatory enzymes, proinflammatory cytokines, and antioxidant enzymes in macrophages. Furthermore, EPCs and NEPCs lowered the viability of colon cancer cells, while normal colon cells were not affected. Additionally, EPCs and NEPCs induced cellular apoptosis and led to G0/G1 cell cycle arrest in HCT116 cells. Overall, these results provide a rationale for future animal and human studies designed to examine the anti-inflammatory and chemopreventive capacities of polyphenols-rich components from L. japonica, U. lactuca, and P. tenera.

Structure and Properties of Organogels Prepared from Rapeseed Oil with Stigmasterol.

This work used the natural ingredient stigmasterol as an oleogelator to explore the effect of concentration on the properties of organogels. Organogels based on rapeseed oil were investigated using various techniques (oil binding capacity, rheology, polarized light microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy) to better understand their physical and microscopic properties. Results showed that stigmasterol was an efficient and thermoreversible oleogelator, capable of structuring rapeseed oil at a stigmasterol concentration as low as 2% with a gelation temperature of 5 °C. The oil binding capacity values of organogels increased to 99.74% as the concentration of stigmasterol was increased to 6%. The rheological properties revealed that organogels prepared with stigmasterol were a pseudoplastic fluid with non-covalent physical crosslinking, and the G' of the organogels did not change with the frequency of scanning increased, showing the characteristics of strong gel. The microscopic properties and Fourier transform infrared spectroscopy showed that stigmasterol formed rod-like crystals through the self-assembly of intermolecular hydrogen bonds, fixing rapeseed oil in its three-dimensional structure to form organogels. Therefore, stigmasterol can be considered as a good organogelator. It is expected to be widely used in food, medicine, and other biological-related fields.

A strategy for accurately and sensitively quantifying free and esterified fatty acids using liquid chromatography mass spectrometry.

Fatty acid (FA) composition of foods dictates a diversity of aspects regarding food quality, ranging from product shelf life, sensory properties to nutrition. There is a challenge to quantitate FAs using liquid chromatography-mass spectrometry due to poor ionization efficiency and matrix effects. Here, an isotopic-tagged derivatization strategy was established to accurately and sensitively quantify free and esterified FAs. After derivatization reaction, the detection sensitivity of FAs was remarkably improved and the limit of quantitation was lower than 100 ng/L. The quantitative errors caused by matrix effects were diminished benefiting from isotope-derivatized internal standards. The established quantitation strategy was successfully applied to verify both free and esterified FA contents in meat after different post-harvest procedures, finding that free polyunsaturated FAs increased significantly during freezing process.