Molecular mechanism of miRNA mediated biosynthesis of secondary metabolites in medicinal plants.

Plant secondary metabolites are important raw materials for the pharmaceutical industry, and their biosynthetic processes are subject to diverse and precise regulation by miRNA. The identification of miRNA molecules in medicinal plants and exploration of their mechanisms not only contribute to a deeper understanding of the molecular genetic mechanisms of plant growth, development and resistance to stress, but also provide a theoretical basis for elucidating the pharmacological effects of authentic medicinal materials and constructing bioreactors for the synthesis of medicinal secondary metabolite components. This paper summarizes the research reports on the discovery of miRNA in medicinal plants and their regulatory mechanisms on the synthesis of secondary metabolites by searching the relevant literature in public databases. It summarizes the currently discovered miRNA and their functions in medicinal plants, and summarizes the molecular mechanisms regulating the synthesis and degradation of secondary metabolites. Furthermore, it provides a prospect for the research and development of medicinal plant miRNA. The compiled information contributes to a comprehensive understanding of the research progress on miRNA in medicinal plants and provides a reference for the industrial development of related secondary metabolite biosynthesis.

Advances in militarine: Pharmacology, synthesis, molecular regulation and regulatory mechanisms.

Militarine is the lead member of secondary metabolites found in multiple medicinal plants of the orchid family. It acts as not only an important inhibitor on plant growth, but also functions as the quality marker for medicinal materials. In addition, Militarine has been shown to possess remarkably medicinal value, with a definite potential for finding widespread adoption of treating various diseases, including lung injury, brain nerve injury, cognitive impairment, aging, tumors, inflammation, peptic ulcers, and more. Thus, it can serve as a material carrier for pharmacophore upon, so much so that it probes as natural source of lead compounds in the research and development of medication. The study reported herein makes an overview on the physicochemical properties and pharmacological mechanisms of Militarine compounds, summarizes the biogenic pathways of Militarine and organically integrates the biological characteristics of Militarine with multiple omics techniques. Besides, this review also constructs a regulatory system for the biological accumulation of Militarine around its precursor compounds, characteristic gene elements, key enzymes, important metabolic products, and critical steps and links. Exceptionally, emphasis on the biosynthesis of Militarine under both abiotic and biotic stress, as well as an elaboration of the signaling pathways and critical regulatory mechanisms that govern the metabolic flow of Militarine have been represented accordingly in this paper. These findings are expected to provide reference schemes and theoretical foundations for acquiring high-quality resources of Militarine and advancing its large-scale industrial production, drug development, and clinical applications to comprehensively elucidate the biosynthetic and metabolic pathways.

The role and mechanism of TCM in the prevention and treatment of infectious diseases.

The constant presence of infectious diseases poses an everlasting threat to the entire world. In recent years, there has been an increased attention toward the application of traditional Chinese medicine (TCM) in the treatment of emerging infectious diseases, as it has played a significant role. The aim of this article is to provide a concise overview of the roles and mechanisms of TCM in treating infectious diseases. TCM possesses the ability to modulate relevant factors, impede signaling pathways, and inhibit microbial growth, thereby exhibiting potent antiviral, antibacterial, and anti-inflammatory effects that demonstrate remarkable efficacy against viral and bacterial infections. This article concludes that the comprehensive regulatory features of Chinese herbal medicines, with their various components, targets, and pathways, result in synergistic effects. The significance of Chinese herbal medicines in the context of infectious diseases should not be underestimated; however, it is crucial to also acknowledge their underutilization. This paper presents constructive suggestions regarding the challenges and opportunities faced by Chinese medicines. Particularly, it emphasizes the effectiveness and characteristics of Chinese medicines in the treatment of infectious diseases, specifying how these medicines' active substances can be utilized to target infectious diseases. This perspective is advantageous in facilitating researchers' pharmacological studies on Chinese medicines, focusing on the specific points of action. The mechanism of action of Chinese herbal medicines in the treatment of infectious diseases is comprehensively elucidated in this paper, providing compelling evidence for the superior treatment of infectious diseases through Chinese medicine. This information is favorable for advancing the development of TCM and its potential applications in the field of infectious diseases.

Identification, Biological Function Profiling and Biosynthesis of Secondary Metabolites in Medicinal Orchids.

The secondary metabolites present in medicinal orchids are diverse and possess a vast array of biological activities. They represent valuable raw materials for modern pharmaceuticals and clinical medicine and have tremendous potential for future development. A systematic collation of secondary metabolites' composition and a summary of the biological activities of medicinal orchids represent a crucial step in unlocking the potential of these valuable resources in drug development. Furthermore, such information can provide essential guidance for comprehensively analyzing the pharmacological and therapeutic mechanisms of these valuable herbs in traditional Chinese herbal medicine. This review article presents an overview of the types and main biological functions of the secondary metabolites found in medicinal orchids, as well as the conventional synthesis methods for these compounds. Our aim is to provide a useful reference for future research and the drug development of secondary metabolic products of medicinal orchids.

Application of traditional Chinese medicine in film drug delivery system.

Film drug delivery systems have the advantages of precise administration, simple process and easy portability, compared with other traditional drug delivery systems such as tablets, capsules, syrups, ointments, etc. The traditional Chinese medicine (TCM) are normally developed in four categories of film agent like patch film, coating, spray film and gel film, which are applied to the treatment of oral ulcers, chronic diseases of lower limbs, burns, scalds, gynecological disease and body care. So the TCM film has great research value and prominent market prospect. In this review, we summarized the research progress of the material composition, pharmaceutical production, clinical application and pharmacology mechanism of various TCM film agents. It may provide a comprehensive reference for further development and utilization of TCM film agents.

Dendrobine biosynthesis in Dendrobium nobile in four different habitats is affected by the variations in the endophytic fungal community.

Dendrobium nobile, an epiphytic plant, is a traditional medicinal herb with abundant endophytes. It is unclear whether the variation in the diversity and abundance of endophytes could stimulate the biosynthesis of medicinal compounds in the plant. In this study, we collected fresh stems of D. nobile from four habitats for investigating the fungal community structure, dendrobine content, and environment factors and their correlations. The results indicated no significant difference in endophytic fungal diversity among the habitats; however, different dominant or special endophytic genera were observed in the hosts from different habitats. The altitude was observed to be positively related to the dendrobine content, as the stems collected from the altitude of 692 m exhibited the highest level of dendrobine. Furthermore, the relative abundance of Toxicocladosporium was found to be positively correlated with the altitude and dendrobine content. The epiphytic matrix exhibited a significant negative correlation with the relative abundance of the endophytic fungus Gibberella but did not exhibit any significant correlation with the dendrobine content. The results indicated that the abundance of endophytes in D. nobile was affected by the altitude and epiphytic matrix and that high Toxicocladosporium abundance and high altitude were conducive to dendrobine production.

Mechanisms by Which Traditional Chinese Medicines Influence the Intestinal Flora and Intestinal Barrier.

The effect of a drug on the intestinal flora and the intestinal barrier is an important evaluation index for drug safety and efficacy. Chemical synthetic drugs are widely used due to their advantages of fast efficacy and low doses, but they are prone to cause drug resistance and inhibit proton pumps, which may harm intestinal health. Traditional Chinese medicine (TCM) has been applied clinically for thousands of years, and how TCMs regulate intestinal health to achieve their effects of disease treatment has become a hot research topic that needs to be resolved. This paper reviews the recent research on the effects of TCMs on intestinal microorganisms and the intestinal mucosal barrier after entering the intestine, discusses the interaction mechanisms between TCMs and intestinal flora, and details the repair effect of TCMs on the intestinal mucosal barrier to provide a reference for the development, utilization, and modernization of TCM.

Glucosyloxybenzyl 2-isobutylmalates and phenolic glycosides from the flowers of Bletilla striata.

Four previously undescribed compounds, including three glucosyloxybenzyl 2-isobutylmalates (1-3), one phenolic glycoside (4), along with ten known compounds were isolated from the flowers of Bletilla striata. The structures and absolute configurations of the undescribed compounds were elucidated on the basis of HR-ESIMS, NMR spectroscopy, optical rotation value, and acid hydrolysis experiment. Cytotoxicity of the isolated compounds against A549, HCT-116, and SW1990 cells and protective effects of t-BHP-induced L02 cytotoxic were assayed. The antioxidant activities of the isolated compounds were also evaluated.

Anti-Vibrio dibutyl phthalate from marine-derived Streptomyces sp. S073.

Marine Streptomyces S073 was previously shown to have strong anti-Vibrio activity, and its antibacterial mechanism was proposed to be associated with siderophore-mediated iron competition and other antagonistic agents. In this study, anti-Vibrio compounds produced by S073 were isolated by bioassay-guided fractionation using column chromatography and HPLC, and the target compound in the most active fraction was identified as dibutyl phthalate (DBP) by various spectroscopic analyses, including EI-MS, 1H NMR and 13C NMR. The DBP-producing capacity of S073 was 2.39 mg/L in ISP1 culture media. Pure DBP was demonstrated to have strong inhibitory activity on Vibiro parahaemolyticus growth with an MIC of 31.25 mg/L. When standard DBP was supplemented into the S073 fermentation broth in a gradient method, an additive inhibitory effect on V. parahaemolyticus was observed, indicating the important role of DBP in driving anti-Vibrio activity in S073 metabolites pool. A synergistic additive effect between DBP and florfenicol was observed in the Vibrio inhibition. These results indicate that, to achieve Vibrio-inhibition, S073 exerted multifaceted strategies, which included DBP-mediated antagonism and siderophore-governed iron competition. The application potential of S073 as an aquaculture probiotic was evaluated, and the safety risks associated with the endocrine disruptor attributes of DBP were discussed.

Callus growth kinetics and accumulation of secondary metabolites of Bletilla striata Rchb.f. using a callus suspension culture.

Bletilla striata is an endangered traditional Chinese medicinal plant with multiple uses and a slow regeneration rate of its germplasm resources. To evaluate the callus growth kinetics and accumulation of secondary metabolites (SMs), a callus suspension culture was proven to be a valuable approach for acquiring high yields of medicinal compounds. An effective callus suspension culture for obtaining B. striata callus growth and its SMs was achieved with the in vitro induction of calluses from B. striata seeds. The callus growth kinetics and accumulation of SMs were analyzed using a mathematical model. The resulting callus growth kinetic model revealed that the growth curves of B. striata suspension-cultured calluses were sigmoidal, indicating changes in the growth of the suspension-cultured calluses. Improved Murashige and Skoog callus growth medium was the most favorable medium for B. striata callus formation, with the highest callus growth occurring during the stationary phase of the cultivation period. Callus growth acceleration started after 7 days and thereafter gradually decreased until day 24 of the cultivation period and reached its highest at day 36 period in both the dry weight and fresh weight analyses. The coelonin concentration peaked during the exponential growth stage and decreased afterward during the stationary stage of the callus suspension culture. The maximum content of coelonin (approximately 0.3323 mg/g callus dry weight) was observed on the 18th day of the cultivation cycle, while dactylorhin A and militarine reached the highest concentrations at day 24, and p-hydroxybenzyl alcohol at day 39. This investigation also laid a foundation for a multimathematical model to better describe the accumulation variation of SMs. The production of SMs showed great specificity during callus growth and development. This research provided a well-organized way to increase the accumulation and production of SMs during the scaled-up biosynthesis of calluses in B. striata callus suspension cultures.

Chemical Constituents, Pharmacologic Properties, and Clinical Applications of Bletilla striata.

Bletilla striata is a plant from the Orchidaceae family that has been employed as a traditional Chinese medicine (TCM) for thousands of years in China. Here, we briefly review the published studies of the last 30 years that were related to chemical constituents, pharmacologic activities, and clinical applications of B. striata. Approximately 158 compounds have been extracted from B. striata tubers with clarified molecular structures that were classified as glucosides, bibenzyls, phenanthrenes, quinones, biphenanthrenes, dihydrophenanthrenes, anthocyanins, steroids, triterpenoids, and phenolic acids. These chemicals support the pharmacological properties of hemostasis and wound healing, and also exhibit anti-oxidation, anti-cancer, anti-viral, and anti-bacterial activities. Additionally, various clinical trials conducted on B. striata have demonstrated its marked activities as an embolizing and mucosa-protective agent, and its application for use in novel biomaterials, quality control, and toxicology. It also has been widely used as a constituent of many preparations in TCM formulations, but because there are insufficient studies on its clinical properties, its efficacy and safety cannot be established from a scientific point of view. We hope that this review will provide reference for further research and development of this unique plant.

De Novo assembly, characterization and development of EST-SSRs from Bletilla striata transcriptomes profiled throughout the whole growing period.

Bletilla striata is an endangered orchid that has been used for millennia as a medicinal herb, in cosmetics and as a horticultural plant. To construct the first nucleotide database for this species and to develop abundant EST-SSR markers for facilitating further studies, various tissues and organs of plants in the main developmental stages were harvested for mRNA isolation and subsequent RNA sequencing. A total of 106,054,784 clean reads were generated by using Illumina paired-end sequencing technology. The reads were assembled into 127,261 unigenes by the Trinity package; the unigenes had an average length of 612 bp and an N50 of 957 bp. Of these unigenes, 67,494 (51.86%) were annotated in a series of databases. Of these annotated unigenes, 41,818 and 24,615 were assigned to gene ontology categories and clusters of orthologous groups, respectively. Additionally, 20,764 (15.96%) unigenes were mapped onto 275 pathways using the KEGG database. In addition, 25,935 high-quality EST-SSR primer pairs were developed from the 15,433 unigenes by MISA mining. To validate the accuracy of the newly designed markers, 87 of 100 randomly selected primers were effectively amplified; 63 of those yielded PCR products of the expected size, and 25 yielded products with significant amounts of polymorphism among the 4 landraces. Furthermore, the transferability test of the 25 polymorphic markers was performed in 6 individuals of two closely related genus Phalaenopsis and dendrobium. Which results showed a total of 5 markers can successfully amplified among these populations. This research provides a comprehensive nucleotide database and lays a solid foundation for functional gene mining and genomic research in B. striata. The developed EST-SSR primers could facilitate phylogenetic studies and breeding.

The guanine nucleotide-binding protein α subunit protein ChGnaq positively regulates Hsc70 transcription in Crassostrea hongkongensis.

Gnaq, one of Guanine nucleotide-binding protein α subunits, was isolated from cellular nucleus extracts of oyster Crassostrea hongkongensis gills with biotin-labeled ChHsc70 promoter by means of DNA-affinity purification, and preliminarily identified with mass spectrometry analysis. ChGnaq mRNA depletion by RNAi technique led to clear reduction in ChHsc70 mRNA expression of C. hongkongensis hemocytes. Correspondently, ChGnaq over-expression in heterologous HEK293T cells correlated with elevated expression activation of ChHsc70 promoter. Quantitative real time PCR analysis showed that both ChHsc70 and ChGnaq transcriptions were responsive to external physical/chemical stresses by heat, CdCl2 and NP. This suggested a plausible association between ChHsc70 and ChGnaq in the stress-induced genetic regulatory pathway. This study discovered a positively regulatory role of ChGnaq in controlling ChHsc70 transcription of C. hongkongensis, and conduced to a better understanding of the regulatory mechanisms in control of Hsc70 transcription.

Construction of full-length cDNA library and development of EST-derived simple sequence repeat (EST-SSR) markers in Senecio scandens.

Senecio scandens Buch.-Ham. ex D. Don (Compositae) is a crucial source of Chinese traditional medicine with antibacterial properties. We constructed a cDNA library and obtained expressed sequence tags (ESTs) to show the distribution of gene ontology annotations for mRNAs, using an individual plant with superior antibacterial characteristics. Analysis of comparative genomics indicates that the putative uncharacterized proteins (21.07%) might be derived from "molecular function unknown" clones or rare transcripts. Furthermore, the Compositae had high cross-species transferability of EST-derived simple sequence repeats (EST-SSR), based on valid amplifications of 206 primer pairs developed from the newly assembled expressed sequence tag sequences in Artemisia annua L. Among those EST-SSR markers, 52 primers showed polymorphic amplifications between individuals with contrasting diverse antibacterial traits. Our sequence data and molecular markers will be cost-effective tools for further studies such as genome annotation, molecular breeding, and novel transcript profiles within Compositae species.

Sequencing and comparative genomics analysis in Senecio scandens Buch.-Ham. Ex D. Don, based on full-length cDNA library.

Senecio scandens Buch.-Ham. ex D. Don, an important antibacterial source of Chinese traditional medicine, has a widespread distribution in a few ecological habitats of China. We generated a full-length complementary DNA (cDNA) library from a sample of elite individuals with superior antibacterial properties, with satisfactory parameters such as library storage (4.30 × 106 CFU), efficiency of titre (1.30 × 106 CFU/mL), transformation efficiency (96.35%), full-length ratio (64.00%) and redundancy ratio (3.28%). The BLASTN search revealed the facile formation of counterparts between the experimental sample and Arabidopsis thaliana in view of high-homology cDNA sequence (90.79%) with e-values <1e - 50. Sequence similarities to known proteins indicate that the entire sequences of the full-length cDNA clones consist of the major of functional genes identified by a large set of microarray data from the present experimental material. For other Compositae species, a large set of full-length cDNA clones reported in the present article will serve as a useful resource to facilitate further research on the transferability of expressed sequence tag-derived simple sequence repeats (EST-SSR) development, comparative genomics and novel transcript profiles.

[Microbial biomass and its correlations with carbon, nitrogen, and phosphorus in the sediments of Taihu Lake].

To explore the responses and feedbacks of the microbes in the sediments of Taihu Lake to the sediment nutrients, an investigation was made on the microbial biomass carbon (MB(C)), microbial biomass nitrogen (MB(N)), microbial biomass phosphorus (MB(P)), and their correlations with the total organic carbon (TOC), total nitrogen (TN), and total phosphorus (TP) in the sediments. The microbial biomass in the sediments was 184.66 mg x kg(-1), being higher at the lakeside than in the mid-lake region. The MB(C) was higher in the western coastal region, Zhushan Bay, and Meiliang Bay, with an average of 127.57 mg x kg(-1), MB(N) was higher in Meiliang Bay, Gonghu Bay, mid-lake region close to Meiliang Bay and Gonghu Bay, and eastern costal region, with an average of 19.25 mg x kg(-1), and MB(P) was higher in the eastern region and parts of the mid-lake region, with an average was 19.09 mg x kg(-1). The TOC high value zone (> or = 2.30 g x kg(-1)) was mainly in Zhushan Bay, western coastal region, Meiliang Bay, and Gonghu Bay, with an average of 1.59 g x kg(-1), TN high value zone (> or = 0.30 g x kg(-1)) was mainly in the Gonghu Bay, Meiliang Bay, Zhushan Bay, and western costal region, with an average of 0.21 g x kg(-1), and TP high value zone (> or = 1.20 g x kg(-1)) was mainly in the eastern coastal region and parts of the mid-lake region, with an average of 0.55 g x kg(-1). The TOC/TN ratio in the sediments was 7-19, with an average of 8.97, which showed that the organic substances in the sediments had obvious dual sources, among which, terrestrial organisms were mainly in the west side of the lake. The microbial biomass in the sediments was significantly positively correlated with sediment TOC and TN but had less correlation with sediment TP, and the MB(C)/MB(N) was significantly correlated with sediment TOC/TN, suggesting that the microbes in the sediments of Taihu Lake were mainly affected by the sediment TOC and TN, and the changes of the TOC/TN had significant effects on the microbial community structure.

S-Adenosylmethionine induces BldH and activates secondary metabolism by involving the TTA-codon control of bldH expression in Streptomyces lividans.

In the present study, a mechanism for S-adenosylmethionine (SAM) to promote secondary metabolism was characterized in terms of bldH sl) expression in Streptomyces lividans. A previous study demonstrated that SAM, on application at 2 microM, induces the transcription of the strR promoter (strRp), which originates from Streptomyces griseus, in S. lividans. An inactivation study verified that bldH sl is essential to strRp transcription in S. lividans and it was demonstrated that the effects of SAM on the induction of strRp activity, on the transcription of redZ and actII-orf4, and on antibiotic production were compromised when the unique leucine TTA-codon of bldH sl was changed to TTG. Western blot analysis revealed that SAM supplementation enhances the expression of bldH sl when the TTA-codon was intact but not when the TTG replacement was provided. This study validates the involvement of BldH sl in the potentiating effect of SAM on the antibiotic production and substantiates that SAM controls the expression of bldH sl through the TTA-codon control in translating bldH mRNA. It is argued here that the intracellular SAM-level modulates the maturation of bldA, which encodes the UUA-codon tRNA and controls secondary metabolism in S. lividans.