The Combination of Bioactive Herbal Compounds with Biomaterials for Regenerative Medicine.

Regenerative medicine aims to restore the function of diseased or damaged tissues and organs by cell therapy, gene therapy, and tissue engineering, along with the adjunctive application of bioactive molecules. Traditional bioactive molecules, such as growth factors and cytokines, have shown great potential in the regulation of cellular and tissue behavior, but have the disadvantages of limited source, high cost, short half-life, and side effects. In recent years, herbal compounds extracted from natural plants/herbs have gained increasing attention. This is not only because herbal compounds are easily obtained, inexpensive, mostly safe, and reliable, but also owing to their excellent effects, including anti-inflammatory, antibacterial, antioxidative, proangiogenic behavior and ability to promote stem cell differentiation. Such effects also play important roles in the processes related to tissue regeneration. Furthermore, the moieties of the herbal compounds can form physical or chemical bonds with the scaffolds, which contributes to improved mechanical strength and stability of the scaffolds. Thus, the incorporation of herbal compounds as bioactive molecules in biomaterials is a promising direction for future regenerative medicine applications. Herein, an overview on the use of bioactive herbal compounds combined with different biomaterial scaffolds for regenerative medicine application is presented. We first introduce the classification, structures, and properties of different herbal bioactive components and then provide a comprehensive survey on the use of bioactive herbal compounds to engineer scaffolds for tissue repair/regeneration of skin, cartilage, bone, neural, and heart tissues. Finally, we highlight the challenges and prospects for the future development of herbal scaffolds toward clinical translation. Overall, it is believed that the combination of bioactive herbal compounds with biomaterials could be a promising perspective for the next generation of regenerative medicine.

Metabolomics analysis reveals the metabolite profiles of Rheum tanguticum grown under different altitudinal gradients.

BACKGROUND: Plant growth and quality are often affected by environmental factors, including geographical location, climate, and soil. In this study, we describe the effect of altitudinal differences on the growth and active ingredients in Rheum tanguticum Maxim. ex Balf. (R. tanguticum), a traditional Chinese medicinal herb known for its laxative properties. RESULTS: The results showed that plants grown at lower altitudes had better growth performances than those in higher altitude areas. The yield varied by 2.45-23.68 times with altitude, reaching a maximum of 102.01 t/ha. In addition, total anthraquinone and total sennoside contents decreased with increasing altitude, whereas total tannins increased with increasing altitude. The total anthraquinone content of the indicator compound reached 5.15% at five experimental sites, which exceeded the Chinese Pharmacopoeia standard by 70.87%. The content of the other two categories of active ingredients reached a maximum value of 0.94% (total sennosides) and 2.65% (total tannins). Redundancy analysis revealed that annual rainfall, annual average temperature, annual sunshine hours, and pH significantly affected growth and active ingredients. Moreover, key metabolites, such as flavonoids, amino acids and their derivatives, phenolic acids, lipids, and terpenes, were differentially expressed between samples from low- and high-altitude cultivation areas. These metabolites were enriched in the flavonoid and flavonol biosynthetic pathway and the monoterpene biosynthetic pathway. CONCLUSIONS: These results suggest that high anthraquinone content was observed in the lowest-latitude cultivation area due to low rainfall and alkaline soil pH. Key metabolites were significantly upregulated in high-latitude cultivation areas. These results provide a scientific basis for quality control and the systematic cultivation of R. tanguticum.

Fabrication of antibacterial poly (L-lactic acid)/tea polyphenol blend films via reactive blending using SG copolymer.

Poly (L-lactic acid) (PLLA) composite materials with both excellent antibacterial properties and mechanical properties are highly desirable for both food packaging and biomedical applications. However, a facile method to prepare transparent PLLA composite films with both excellent antibacterial and mechanical properties is still lacking. In this work, blend films based on PLLA, tea polyphenols (TP) and poly (styrene-co-glycidyl methacrylate) (SG) copolymers (PLLA/TP/SG) were prepared by melt blending using twin screw extruder. The blend films showed high transparency with a brownish color originated from tea polyphenols. Both SEM and DSC analyses confirmed that the blends are thermodynamically compatible. GPC and mechanical assessments demonstrated that the PLLA/TP binary blends exhibit reduced molecular weight and compromised mechanical properties, compared to neat PLLA. However, incorporating SG copolymer resulted in increased molecular weight and improved mechanical properties for the PLLA/TP/SG blends. The FT-IR spectra exhibited a shift to lower wavenumber for the absorption peak associated with the benzene ring on TPs after blending with PLLA and SG, indicating the occurrence of transesterification between PLLA and TP. Plate coating studies revealed that the PLLA/TP/SG blends with TP incorporation at 5 wt% exhibited a bacteriostatic rate of 99.99 % against Staphylococcus aureus and Escherichia coli. Overall, our study reveals that the PLLA/TP/SG blend films exhibit excellent antibacterial properties coupled with good mechanical properties, rendering them a promising candidate for antibacterial packaging materials.

Transcriptomic and proteomic investigation of the ameliorative effect of total polyphenolic glycoside extract on hepatic fibrosis in Lamiophlomis rotata Kudo via the AGE/RAGE pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: During the regression of liver fibrosis, a decrease in hepatic stellate cells (HSCs) can occur through apoptosis or inactivation of activated HSCs (aHSCs). A new approach for antifibrotic therapy involves transforming hepatic myofibroblasts into a quiescent-like state. Lamiophlomis rotata (Benth.) Kudo (L. rotata), an orally available Tibetan herb, has traditionally been used to treat skin disease, jaundice, and rheumatism. In our previous study, we found that the total polyphenolic glycoside extract of L. rotata (TPLR) promotes apoptosis in aHSCs for the treatment of hepatic fibrosis. However, whether TPLR induces aHSCs to become inactivated HSCs (iHSCs) is unclear, and the underlying mechanism remains largely unknown. PURPOSE: This study aimed to examine the impact of TPLR on the phenotypes of hepatic stellate cells (HSCs) during the regression of liver fibrosis and explore the potential mechanism of action. METHODS: The effect of TPLR on the phenotypes of hepatic stellate cells (HSCs) was assessed using immunofluorescence (IF) staining, reverse transcription-polymerase chain reaction (RT-PCR), and Western blotting. Transcriptomic and proteomic methods were employed to identify the main signaling pathways involved. Based on the omics results, the likely mechanism of TPLR on the phenotypes of aHSCs was confirmed through overexpression and knockdown experiments in TGF-ß1-activated LX-2 cells. Using a CCl4-induced liver fibrosis mouse model, we evaluated the anti-hepatic fibrosis effect of TPLR and explored its potential mechanism based on omics findings. RESULTS: TPLR was found to induce the differentiation of aHSCs into iHSCs by significantly decreasing the protein expression of α-SMA and Desmin. Transcriptomic and proteomic analyses revealed that the AGE/RAGE signaling pathway plays a crucial role in the morphological transformation of HSCs following TPLR treatment. In vitro experiments using RAGE overexpression and knockdown demonstrated that the mechanism by which TPLR affects the phenotype of HSCs is closely associated with the RAGE/RAS/MAPK/NF-κB axis. In a model of liver fibrosis, TPLR obviously inhibited the generation of AGEs and alleviated liver tissue damage and fibrosis by downregulating RAGE and its downstream targets. CONCLUSION: The AGE/RAGE axis plays a pivotal role in the transformation of activated hepatic stellate cells (aHSCs) into inactivated hepatic stellate cells (iHSCs) following TPLR treatment, indicating the potential of TPLR as a therapeutic agent for the management of liver fibrosis.

Trophic relationships between protists and bacteria and fungi drive the biogeography of rhizosphere soil microbial community and impact plant physiological and ecological functions.

Rhizosphere microorganisms play a vital role in enhancing plant health, productivity, and the accumulation of secondary metabolites. Currently, there is a limited understanding of the ecological processes that control the assembly of community. To address the role of microbial interactions in assembly and for functioning of the rhizosphere soil microbiota, we collected rhizosphere soil samples from Anisodus tanguticus on the Tibetan Plateau spanning 1500 kilometers, and sequenced the bacteria, fungi, archaea, and protist communities. We observed a significant but weak distance-decay relationship in the microbial communities of rhizosphere soil. Our comprehensive analysis of spatial, abiotic, and biotic factors showed that trophic relationships between protists and bacteria and fungi predominantly influenced the alpha and beta diversity of bacterial, fungal, and protistan communities, while abiotic factors had a greater impact on archaeal communities, including soil pH, available phosphorus, total phosphorus and mean annual temperature. Importantly, microbial interactions had a more significant influence on Anisodus tanguticus physiological and ecological functions compared to individual microorganisms. Network analyses revealed that bacteria occupy a central position of the co-occurrence network and play a crucial role of connector within this community. The addition of protists increased the stability of bacterial, fungal, and archaeal networks. Overall, our findings indicate that trophic relationships play an important role in assembly and for functioning of the rhizosphere soil microbiota. Bacterial communities serve as a crucial link between different kingdoms of microorganisms in the rhizosphere community. These findings help us to fully harness the beneficial functions of rhizosphere microorganisms for plants and achieve sustainable use of biological resources.

A core root bacteria contribute to plant growth and anisodine accumulation of Anisodus tanguticus.

BACKGROUND: Although it is well recognized that core root microorganisms contribute to plant health and productivity, little is known about their role to the accumulation of secondary metabolites. The roots of Anisodus tanguticus, a traditional herbal medication utilized by Tibetan medicine, are rich in tropane alkaloids. We collected wild A. tanguticus populations throughout a 1500 km transect on the Qinghai-Tibetan Plateau. RESULTS: Our results showed that despite sampling at a distance of 1500 km, the root of A. tanguticus selectively recruits core root bacteria. We obtained 102 root bacterial core OTUs, and although their number only accounted for 2.99% of the total, their relative abundance accounted for 73% of the total. Spearman correlation and random forest analyses revealed that the composition of core root microbiomes was related to anisodine contents, aboveground biomass and nitrogen contents of Anisodus tanguticus. Among them, the main role is played by Rhizobacter, Variovorax, Polaromonas, and Mycobacterium genus that are significantly enriched in roots. Functional prediction by FAPROTAX showed that nitrogen-cycling microorganisms and pathogenic bacteria are strongly associated with anisodine contents, aboveground biomass and nitrogen contents of Anisodus tanguticus. CONCLUSIONS: Our findings show that the root selectively recruits core root bacteria and revealed that the core microbiomes and microbial functions potentially contributed to the anisodine contents, aboveground biomass and nitrogen contents of the plant. This work may increase our understanding of the interactions between microorganisms and plants and improve our ability to manage root microbiota to promote sustainable production of herbal medicines.

Soluble Protein Content, Bioactive Compounds and the Antioxidant Activity in Seeds of Ten Rheum tanguticum Lines from Qinghai-Tibet Plateau.

Rheum tanguticum (Rh. tanguticum) is a Chinese medicinal plant traditionally used in the treatment of constipation. As a byproduct, the seeds of this plant are rich in nutrients and phytochemicals. This study aimed to determine and assess seed germination ability, seed physical characteristics, soluble protein content, chemical constituents and antioxidant capacity from different breeding lines, to promote the development and utilization of seed resources. Significant differences were observed for the soluble protein content and antioxidant assays among the ten lines. The contents of aloe-emodin, rhein and catechins accumulated in seeds were extremely low and significantly different from those in roots. In contrast, emodin and chrysophanol were abundant in seeds, and significant differences were observed between seeds and roots. It was found that associations between gallic acid and catechins were not significant for either soluble protein or antioxidant capacity. There was a significantly positive correlation between the contents of four anthraquinones (aloe-emodin, rhein, emodin and chrysophanol) and soluble protein. Seeds have potent antioxidative capacity and relatively high levels of soluble protein content. The rich chemical composition of seeds can be widely used in the medical industry for further development.

Early Bolting, Yield, and Quality of Angelica sinensis (Oliv.) Diels Responses to Intercropping Patterns.

Intercropping is a sustainable method for cultivating medicinal herbs since it requires lower dependence on chemical fertilizers than a sole cropping system. In this study, we compared the effects of sole cropping and intercropping on early bolting, yield, and the chemical composition of Angelica sinensis (Oliv.) Diels. Field experiments were conducted, in 2018 and in 2019, using different cropping systems including sole cropping of A. sinensis (AS), sole cropping of Vicia faba (VF), and intercropping (without fertilization) at three ratios: one row of A. sinensis + three rows of V. faba, AS/VF (1:3), two rows of A. sinensis + two rows V. faba, AS/VF (2:2), three rows of A. sinensis + one row V. faba, AS/VF (3:1). The effect of each cropping system was evaluated by measuring the dry biomass of V. faba and the dry biomass, ferulic acid content, and essential oil content and composition of A. sinensis. The early bolting rate of A. sinensis was significantly lower in the intercropping system as compared with that in a sole cropping system. The AS/VF (3:1) intercropping pattern resulted in an optimal yield and the highest ferulic acid content of A. sinensis, highest dry biomass of V. faba, and highest land equivalent ratio (LER). Additionally, the A. sinensis was more aggressive (the aggressivity value of A. sinensis was positive, and its competitive ratio was >1) under AS/VF (3:1) intercropping pattern, and it dominated over V. faba (which had negative aggressivity values and a competitive ratio of <1) under AS/VF (3:1) intercropping pattern. Ligustilide was the most dominant component of the essential oil of A. sinensis, regardless of the cropping system; however, the chemical component of essential oil was not influenced by intercropping patterns. Overall, the AS/VF (3:1) intercropping pattern without fertilization was the most productive, with the highest LER and ferulic acid content. These data indicate that intercropping can serve as an alternative for reducing the use of chemical fertilizers and intercropping also decreases the early bolting rate of A. sinensis, thus, enabling its sustainable production.

Multivariate Statistical Analysis of Metabolites in Anisodus tanguticus (Maxim.) Pascher to Determine Geographical Origins and Network Pharmacology.

Anisodus tanguticus (Maxim.) Pascher, has been used for the treatment of septic shock, analgesia, motion sickness, and anesthesia in traditional Tibetan medicine for 2,000 years. However, the chemical metabolites and geographical traceability and their network pharmacology are still unknown. A total of 71 samples of A. tanguticus were analyzed by Ultra-Performance Liquid Chromatography Q-Exactive Mass Spectrometer in combination with chemometrics developed for the discrimination of A. tanguticus from different geographical origins. Then, network pharmacology analysis was used to integrate the information of the differential metabolite network to explore the mechanism of pharmacological activity. In this study, 29 metabolites were identified, including tropane alkaloids, hydroxycinnamic acid amides and coumarins. Principal component analysis (PCA) explained 49.5% of the total variance, and orthogonal partial least-squares discriminant analysis (OPLS-DA) showed good discrimination (R2Y = 0.921 and Q2 = 0.839) for A. tanguticus samples. Nine differential metabolites accountable for such variations were identified through variable importance in the projection (VIP). Through network pharmacology, 19 components and 20 pathways were constructed and predicted for the pharmacological activity of A. tanguticus. These results confirmed that this method is accurate and effective for the geographic classification of A. tanguticus, and the integrated strategy of metabolomics and network pharmacology can explain well the "multicomponent--multitarget" mechanism of A. tanguticus.

Network pharmacology combined with molecular docking to explore the potential mechanisms for the antioxidant activity of Rheum tanguticum seeds.

BACKGROUND: Rheum tanguticum (R. tanguticum) is an edible and medicinal plant that exhibits high antioxidant activity. The purpose of the present study was to investigate the bioactive components of its seeds and the potential mechanisms of antioxidant activity to provide a foundation for further developmental work on R. tanguticum seeds as a functional food. METHODS: In this study, the antioxidant activities of R. tanguticum seeds were measured using DPPH, ABTS and FRAP assays. LC-Q-TOF/MS was used to identify the active compounds in the seeds, and Swiss Target Prediction was used to identify their potential targets. The DisGENET, DrugBank, OMIM and GeneCard databases were used to search for antioxidant-related targets. RESULTS: The component-target-pathway network was constructed and included 5 compounds and 9 target genes. The hub genes included ESR1, APP, MAPK8, HSP90AA1, AKT1, MMP2, PTGS2, TGFB1 and JUN. The antioxidant activity signaling pathways of the compounds for the treatment of diseases were the cancer signaling pathway, estrogen signaling pathway, colorectal cancer signaling pathway, MAPK signaling pathway, etc. Molecular docking revealed that the compounds in R. tanguticum seeds could inhibit potential targets (AKT1, ESR1 and PTGS2). CONCLUSION: Molecular docking studies revealed that the binding energy score between liriodenine and PTGS2 was the highest (8.16), followed by that of chrysophanol (7.10). This result supports the potential for PTGS2-targeted drug screening and design.

MaxEnt Modeling to Predict the Current and Future Distribution of Pomatosace filicula under Climate Change Scenarios on the Qinghai-Tibet Plateau.

As an important Tibetan medicine and a secondary protected plant in China, Pomatosace filicula is endemic to the country and is mainly distributed in the Qinghai-Tibet Plateau (QTP). However, global climate change caused by greenhouse gas emissions might lead to the extinction of P. filicula. To understand the potential spatial distribution of P. filicula in future global warming scenarios, we used the MaxEnt model to simulate changes in its suitable habitat that would occur by 2050 and 2070 using four representative concentration pathway (RCP) scenarios and five global climate models. The results showed that the QTP currently contains a suitable habitat for P. filicula and will continue to do so in the future. Under the RCP2.6 scenario, the suitable habitat area would increase by 2050 but shrink slightly by 2070, with an average reduction of 2.7%. However, under the RCP8.5 scenario, the area of unsuitable habitat would expand by an average of 54.65% and 68.20% by 2050 and 2070, respectively. The changes in the area of suitable habitat under the RCP4.5 and RCP6.0 scenarios were similar, with the unsuitable area increasing by approximately 20% by 2050 and 2070. Under these two moderate RCPs, the total suitable area in 2070 would be greater than that in 2050. The top three environmental factors impacting the habitat distribution were altitude, annual precipitation (BIO12) and annual temperature range (BIO7). The cumulative contribution rate of these three factors was as high as 82.8%, indicating that they were the key factors affecting the distribution and adaptability of P. filicula, P. filicula grows well in damp and cold environments. Due to global warming, the QTP will become warmer and drier; thus, the growing area of P. filicula will move toward higher elevations and areas that are humid and cold. These areas are mainly found near the Three-River Region. Future climate change will aggravate the deterioration of the P. filicula habitat and increase the species' survival risk. This study describes the distribution of P. filicula and provides a basis for the protection of endangered plants in the QTP.

Above- and below-ground resource acquisition strategies determine plant species responses to nitrogen enrichment.

BACKGROUND AND AIMS: Knowledge of plant resource acquisition strategies is crucial for understanding the mechanisms mediating the responses of ecosystems to external nitrogen (N) input. However, few studies have considered the joint effects of above-ground (light) and below-ground (nutrient) resource acquisition strategies in regulating plant species responses to N enrichment. Here, we quantified the effects of light and non-N nutrient acquisition capacities on species relative abundance in the case of extra N input. METHODS: Based on an N-manipulation experiment in a Tibetan alpine steppe, we determined the responses of species relative abundances and light and nutrient acquisition capacities to N enrichment for two species with different resource acquisition strategies (the taller Stipa purpurea, which is colonized by arbuscular mycorrhizal fungi, and the shorter Carex stenophylloides, which has cluster roots). Structural equation models were developed to explore the relative effects of light and nutrient acquisition on species relative abundance along the N addition gradient. KEY RESULTS: We found that the relative abundance of taller S. purpurea increased with the improved light acquisition along the N addition gradient. In contrast, the shorter C. stenophylloides, with cluster roots, excelled in acquiring phosphorus (P) so as to elevate its leaf P concentration under N enrichment by producing large amounts of carboxylate exudates that mobilized moderately labile and recalcitrant soil P forms. The increased leaf P concentration of C. stenophylloides enhanced its light use efficiency and promoted its relative abundance even in the shade of taller competitors. CONCLUSIONS: Our findings highlight that the combined effects of above-ground (light) and below-ground (nutrient) resources rather than light alone (the prevailing perspective) determine the responses of grassland community structure to N enrichment.

Non-target metabolomics revealed the differences between Rh. tanguticum plants growing under canopy and open habitats.

BACKGROUND: Rheum tanguticum (Rh. tanguticum) is an important traditional Chinese medicine plant, "Dahuang", which contains productive metabolites and occupies wide habitats on the Qinghai-Tibet plateau. Plants occupying wide habitats usually vary in phenotypes such as in morphology and metabolism, thereby developing into different ecotypes. Under canopy and open habitats are a pair of dissimilar habitats which possess Rh. tanguticum plants. However, few studies have focused on the effect of habitats on Rh. tanguticum growth, particularly combining morphological and metabolic changes. This study focused on Rh. tanguticum plants growing in under canopy and open habitats where morphology and metabolism changes were quantified using non-target metabolism methods. RESULTS: The obtained results indicated that the two dissimilar habitats led to Rh. tanguticum developing into two distinct ecotypes where the morphology and metabolism were simultaneously changed. Under canopy habitats bred morphologically smaller Rh. tanguticum plants which had a higher level of metabolites (22 out of 31) which included five flavonoids, four isoflavonoids, and three anthracenes. On the other hand, the open habitats produced morphologically larger Rh. tanguticum plants having a higher level of metabolites (9 out of 31) including four flavonoids. 6 of the 31 metabolites were predicted to have effect targets, include 4 represent for under canopy habitats and 2 for open habitats. Totally, 208 targets were connected, among which 42 were communal targets for both under canopy and open habitats represent compounds, and 100 and 66 were unique targets for under canopy superior compounds and open habitats superior compounds, respectively. In addition, aloe-emodin, emodin, chrysophanol, physcion, sennoside A and sennoside B were all more accumulated in under canopy habitats, and among which aloe-emodin, emodin, chrysophanol and physcion were significantly higher in under canopy habitats. CONCLUSIONS: This study determined that Rh. tanguticum growing in under canopy and in open habitats developed into two distinct ecotypes with morphological and metabolic differences. Results of network pharmacology study has indicated that "Dahuang" coming from different habitats, such as under canopy and open habitats, are different in effect targets and thus may have different medicinal use. According to target metabolomics, under canopy habitats may grow better "Dahuang".

The complete chloroplast genome of Tibetan medicine Gentianopsis paludosa.

Gentianopsis paludosa (Mum.) Ma is an important species in Tibetan folk medicine, but its wild populations are shrinking roughly due to the increasing demand for it. Gentianopsis paludosa is presently at risk of over-exploitation, so it needs urgent conservation. Here, we report the complete sequence of the chloroplast genome of G. paludosa. The genome was 51,121 bp in length with 129 genes comprising 84 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The overall GC content of G. paludosa chloroplast genome was is 36.67%. The phylogenomic analysis suggested that G. paludosa forms a clade with species in Halenia and Swertia, indicating that the G. paludosa is more closely related to Halenia and Swertia than that of Gentiana.

The complete chloroplast genome of Bupleurum longicaule var. strictum, an annual herb endemic to China.

Bupleurum species are well-known for their antipyretic, analgesic, gallbladder and other functions so that they are very popular in traditional Chinese medicine. However, to our knowledge, there is no completed chloroplast genome of Bupleurum genus in China. In the present study, we determined the complete chloroplast genome sequences for Bupleurum longicaule var. strictum using IIumina sequencing. Bupleurum longicaule var. strictum is 155,578 bp which is composed of two inverted repeats (IR: 26,257bp), a large single copy region (LSC: 86,977 bp), and a small single-copy region (SSC: 16,087 bp). The overall AT content is 65.32%. The chloroplast genome includes a total of 51 functional genes including 15 protein-coding genes and 36 tRNA. A total of 10 genes were duplicated in the IR regions including seven tRNA and three protein-coding genes. Phylogenetic analysis suggested that B. longicaule var. strictum formed a monophyletic clade.

Nitrogen Fertilizer Levels Affect the Growth and Quality Parameters of Astragalus mongolica.

Owing to overexploitation, wild resources of Astragalus mongolica, a Chinese herbal plant that is widely distributed in the arid and semi-arid areas of Northern China, have gradually become exhausted, and therefore, commercial cultivation is increasingly important to meet the growing demand for astragalus and reduce the pressure on wild populations. Nitrogen level is an important factor that affects the yield and quality of A. mongolica. However, uniform standards for fertilization among production areas have not yet been determined. In this study, the effect of nitrogen fertilizer treatment on the yield and quality of A. mongolica in the Qinghai-Tibet Plateau was explored using a control treatment (no added nitrogen, N0) and five different nutrient levels: 37.5 kg/ha (N1), 75 kg/ha (N2), 112.5 kg/ha (N3), 150 kg/ha (N4), and 187.5 kg/ha (N5). According to grey relational analysis, the optimal nitrogen fertilizer treatment was the N4 level followed by the N5 and N2 levels. Nitrogen fertilizer significantly increased the root biomass, plant height, root length, and root diameter. However, nitrogen fertilization had no significant effect on the content of Astragaloside IV and mullein isoflavone glucoside. The content of ononin and calycosin continually accumulated throughout the growing period. The results showed that the ononin and calycosin content under N4 and N2 is higher than other levels and there is not significantly different between different nitrogen fertilizer levels about them. The content of formononetin decreased gradually with the progression of the growing season. The optimal nitrogen fertilizer treatment for A. mongolica is recommended to be 150 kg/ha and the content of active compounds and yield were observed to reach the maximum in October.

The complete chloroplast genome of Swertia tetraptera and phylogenetic analysis.

Swertia tetraptera, native to the Qinghai-Tibetan Plateau, is an important traditional Chinese medicine. Although researchers have done a lot of work on it, the phylogenetic position of S. tetraptera within Swertia has still not been solved. Chloroplast genome sequences play a significant role in the development of molecular markers in plant phylogenetic and population genetic studies. In present study, we determined the complete chloroplast genome sequences for S. tetraptera using IIumina sequencing. The total length of the complete chloroplast genome of S. tetraptera is 152,840 bp, of which the GC content is 37.95%. The genome encodes 130 functional genes, including 85 protein-coding genes, 37 tRNA, and 8 rRNA. Phylogenetic analysis suggested that S. tetraptera forms monophyletic group with Halenia corniculata which shows closed relationship with the Halenia.

Effects of different nitrogen fertilizer levels on growth and active compounds of rhubarb from Qinghai plateau.

BACKGROUND: Rheum tanguticum Maxim. ex Balf is one of the plants generically known as rhubarb, a culinary vegetable that has long been used as a herbal remedy both in China and Europe. Increasing demand for rhubarb has triggered the overexploitation of Rh. tanguticum. Cultivation is therefore necessary for quality control and protection of wild resources. Nitrogen fertilizer plays an important role in cultivation. This study aimed to explore how nitrogen fertilizer affects the growth and quality of rhubarb on the Qinghai-Tibetan plateau. RESULTS: Nitrogen fertilizer promoted growth but had no significant influence on the active compounds of Rh. tanguticum. Generally, the N2 (150 kg ha-1 ) and N3 (225 kg ha-1 ) levels showed the most improved growth indexes, with no significant differences between them. The growth index and the amounts of eight of the nine studied active compounds in Rh. tanguticum increased from each year to the next and differed among growth stages. The contents of the active compounds were higher at the green stage and lower at the growth stage, which was opposite to the seasonal trends in root dry matter ratio. Gallic acid levels decreased with the growth of the plant. CONCLUSION: The N2 level (150 kg ha-1 ) was the recommended nitrogen fertilizer level in this study. It was revealed that seasonal changes rather than nitrogen fertilizer influenced active compounds in the root of Rh. tanguticum. © 2018 Society of Chemical Industry.

Separation of three polar compounds from Rheum tanguticum by high-speed countercurrent chromatography with an ethyl acetate/glacial acetic acid/water system.

The separation of polar compounds by high-speed countercurrent chromatography is still regarded as a challenge. In this study, an efficient strategy for the separation of three polar compounds from Rheum tanguticum has been successfully conducted by using high-speed countercurrent chromatography. X-5 macroporous resin chromatography was used for the fast enrichment of the target compounds. Then, the target fraction was directly introduced into high-speed countercurrent chromatography for separation using ethyl acetate/glacial acetic acid/water (100:1:100, v/v/v) as the solvent system. Consequently, three polar compounds including gallic acid, catechin, and gallic acid 4-O-ß-d-(6'-O-galloyl) glucoside were obtained with purities higher than 98%. The results showed glacial acetic acid could be such an appropriate regulator for the ethyl acetate/water system. This study provides a reference for the separation of polar compounds from natural products by high-speed countercurrent chromatography.

Efficient Accumulation and In Vitro Antitumor Activities of Triterpene Acids from Submerged Batch--Cultured Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes).

Triterpene acids are among the major bioactive constituents of lucidum. However, submerged fermentation techniques for isolating triterpene acids from G. lucidum have not been optimized for commercial use, and the antitumor activity of the mycelial triterpene acids needs to be further proven. The aim of this work was to optimize the conditions for G. lucidum culture with respect to triterpene acid production, scaling up the process, and examining the in vitro antitumor activity of mycelial triterpene acids. The key conditions (i.e., initial pH, fermentation temperature, and rotation speed) were optimized using response surface methodology, and the in vitro antitumor activity was evaluated using the MTT method. The optimum key fermentation conditions for triterpene acid production were pH 6.0; rotation speed, 161.9 rpm; and temperature, 30.1°C, resulting in a triterpene acid yield of 291.0 mg/L in the validation experiment in a 5-L stirred bioreactor; this yield represented a 70.8% increase in titer compared with the nonoptimized conditions. Furthermore, the optimized conditions were then successfully scaled up to a production scale of 200 L, and a triterpene productivity of 47.9 mg/L/day was achieved, which is, to our knowledge, the highest reported in the large-scale fermentation of G. lucidum. In addition, the mycelial triterpene acids were found to be cytotoxic to the SMMC-7721 and SW620 cell lines in vitro. Chemical analysis showed that the key active triterpene acid compounds, ganoderic acids T and Me, predominated in the extract, at 69.2 and 41.6 mg/g, respectively. Thus, this work develops a simple and feasible batch fermentation technique for the large-scale production of antitumor triterpene acids from G. lucidum.