Terpene Lactones Research Articles

Differences in in vivo absorption of flavone glycosides, flavone aglycones and terpene lactones under different dosage forms and physiological conditions.

Ginkgo biloba L. extract (GBE) oral preparations have been used for many years in the prevention and treatment of cardiovascular and cerebrovascular diseases, and the main active ingredients are flavonoids and terpene lactones. Among them, the oral absorption of the prototype components of flavonoid glycosides into the blood needs to be further clarified, and the differences in the oral absorption of different components in GBE by different dosage forms and physiological conditions are not clear yet. To clarify the oral absorption of the prototype flavonoid glycosides in vivo, and to further explore the differences in the oral absorption of various active compounds under different oral dosage forms and dietary conditions. Firstly, the target compounds were selected based on the characteristic chromatogram of GBE and literature. Then, the content differences of three different oral GBE preparations were studied, and their pharmacokinetics (PK) were compared. Finally, the PK differences of the preparations with better oral absorption under different dietary conditions were studied. Five flavonoid glycosides, three aglycones and four terpene lactones were selected as the research objects. The content determination results of GBE tablets, guttate pills and tinctures showed that the content of several components especially flavonoid glycosides in the tincture was higher than that of the other two preparations. After oral administration of these three preparations, the PK study showed different results from previous studies. The PK behavior of flavonoid glycosides was also determined at the same time as flavonoid glycosides and terpene lactones. and the bioavailability of flavonoid glycosides in the tincture was higher than that of the other two preparations. PK results of fasting and non-fasting showed that taking GBE tincture on an empty stomach increased the absorption of various compounds, especially flavonoid glycosides. However, due to the existence of food residues in the gastrointestinal tract, the oral bioavailability of flavonoid glycosides was significantly improved. This study discussed the differences in the content and oral absorption of active compounds in different oral preparations of GBE, clarified the in vivo absorption of flavonoid glycosides prototype, as well as the influence of diet on the PK of active compounds, which has certain guiding significance for the clinical application of GBE oral preparations.

Pharmacological activities and effective substances of the component-based Chinese medicine of Ginkgo biloba leaves based on serum pharmacochemistry, metabonomics and network pharmacology.

As a potential drug candidate for the treatment of hypertension and complications, it is speculated that the component-based Chinese medicine of Ginkgo biloba leaves (GBCCM) which mainly composed of flavonoid aglycones (FAs) and terpene lactones (TLs) may have different pharmacological effects at different doses or ratios. Taking the normal mice as the study object, metabonomics was conducted by giving different doses of GBCCM. Based on the components of GBCCM absorbed into the blood, the network pharmacological prediction was carried out. By integrating the results of metabonomics and network pharmacology, predict the possible pharmacological effects of GBCCM and conduct experimental verification. It was found that eight of the 19 compounds in GBCCM could be absorbed into the blood. GBCCM mainly affected the signal pathways of unsaturated fatty acid, pyruvate, bile acid, melanin and stem cells. It was speculated that GBCCM might have activities such as lowering blood pressure, regulating stem cell proliferation and melanogenesis. By establishing the models of mushroom tyrosinase, rat bone marrow mesenchymal stem cells (BMSCs) and spontaneously hypertensive rats (SHRs), we found that FAs and TLs showed synergistic effect in hypertension and tyrosinase models, and the optimal ratio was 3:2 (4.4mg/kg) and 1:1 (0.4mg/ml), respectively. As effective substances, FAs significantly promoted the proliferation of rat BMSCs on the third and fifth days at the concentration of 0.2μg/ml (p < 0.05). GBCCM showed a variety of pharmacological effects at different doses and ratios, which provided an important reference for the druggability of GBCCM.

Quality evaluation of ginkgo biloba leaves based on non-targeted metabolomics and representative ingredient quantification

Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) combined with multivariate statistical analysis was applied to the study of plant metabolomics to reveal the factors affecting the content of ginkgo leaf compounds. As a follow-up analysis, the terpene lactones and ginkgolic acids were quantified simultaneously using ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-QqQ-MS/MS), and subsequently total flavonol glycosides were quantified by high-performance liquid chromatography (HPLC). The results revealed that a total of 52 compounds were potentially identified by establishing a database, and 10 compounds were verified by reference standards; terpene lactones, ginkgolic acids, and flavonoids were the differential compounds; and ginkgolide A was identified as an important indicator compound for tree age. In addition, quantitative analysis showed that the contents of total flavonol glycosides and terpene lactones were highest during April and August in young ginkgo leaves, and differed based on origin. In summary, numerous compounds were rapidly detected by liquid chromatography coupled with MS, the ginkgo leaf samples were compared, and the differential metabolites were screened out. The content changing rules of the target compounds in ginkgo leaves from different regions with different tree ages and harvesting periods were clarified.

Orally Administered Ginkgolide C Attenuates DSS-Induced Colitis by Maintaining Gut Barrier Integrity, Inhibiting Inflammatory Responses, and Regulating Intestinal Flora.

Ulcerative colitis (UC), one of the foremost common forms of inflammatory bowel disease, poses a serious threat to human health. Currently, safe and effective treatments are not available. This study investigated the protective effect of ginkgolide C (GC), a terpene lactone extracted from Ginkgo biloba leaves, on UC and its underlying mechanism. The results showed that GC remarkably mitigated the severity of DSS-induced colitis in mice, as demonstrated by decreased body weight loss, reduced disease activity index, mitigated tissue damage, and increased colon length. Furthermore, GC inhibited DSS-induced hyperactivation of inflammation-related signaling pathways (NF-κB and MAPK) to reduce the production of inflammatory mediators, thereby mitigating the inflammatory response in mice. GC administration also restored gut barrier function by elevating the number of goblet cells and boosting the levels of tight junction-related proteins (claudin-3, occludin, and ZO-1). In addition, GC rebalanced the intestinal flora of DSS-treated mice by increasing the diversity of the flora, elevating the abundance of beneficial bacteria, such as Lactobacillus and Allobaculum, and decreasing the abundance of harmful bacteria, such as Bacteroides, Oscillospira, Ruminococcus, and Turicibacter. Taken together, these results suggest that GC administration effectively alleviates DSS-induced colitis by inhibiting the inflammatory response, maintaining mucosal barrier integrity, and regulating intestinal flora. This study may provide a scientific basis for the rational use of GC in preventing colitis and other related diseases.

Treatment of Ginkgo biloba with Exogenous Sodium Selenite Affects Its Physiological Growth, Changes Its Phytohormones, and Synthesizes Its Terpene Lactones

Ginkgolide is a unique terpenoid natural compound in Ginkgo biloba, and it has an important medicinal value. Proper selenium has been reported to promote plant growth and development, and improve plant quality, stress resistance, and disease resistance. In order to study the effects of exogenous selenium (Se) on the physiological growth and the content of terpene triolactones (TTLs) in G. biloba seedlings, the seedlings in this work were treated with Na2SeO3. Then, the physiological indexes, the content of the TTLs, and the expression of the related genes were determined. The results showed that a low dose of Na2SeO3 was beneficial to plant photosynthesis as it promoted the growth of ginkgo seedlings and increased the root to shoot ratio. Foliar Se application significantly increased the content of soluble sugar and protein and promoted the content of TTLs in ginkgo leaves; indeed, it reached the maximum value of 7.95 mg/g in the ninth week, whereas the application of Se to the roots inhibited the synthesis of TTLs. Transcriptome analysis showed that foliar Se application promoted the expression levels of GbMECPs, GbMECT, GbHMGR, and GbMVD genes, whereas its application to the roots promoted the expression of GbDXS and GbDXR genes. The combined analysis results of metabolome and transcriptome showed that genes such as GbDXS, GbDXR, GbHMGR, GbMECPs, and GbCYP450 were significantly positively correlated with transcription factors (TFs) GbWRKY and GbAP2/ERF, and they were also positively correlated with the contents of terpene lactones (ginkgolide A, ginkgolide B, ginkgolide M, and bilobalide). Endogenous hormones (MeJA-ILE, ETH, and GA7) were also involved in this process. The results suggested that Na2SeO3 treatment affected the transcription factors related to the regulation of endogenous hormones in G. biloba, and further regulated the expression of genes related to the terpene synthesis structure, thus promoting the synthesis of ginkgo TTLs.

Phytochemical Compounds, Antioxidant, and Digestive Enzymes Inhibitory Activities of Different Fractions from Ginkgo biloba L. Nut Shells

This study was designed to investigate the phytochemical compounds, antioxidant, and digestive enzymes inhibitory activities of the free (F), esterified (E), and insoluble-bound (IB) fractions from Ginkgo biloba L. nut shells. Results showed that a total of twelve compounds were detected in G. biloba nut shells by using UHPLC-ESI-HRMS/MS, including two kinds of organic acids, three kinds of phenolic acids, three kinds of flavonoids, and four kinds of terpene lactones. The F fraction contained all identified compounds and had the highest contents of the total phenolics and total flavonoids. All of the three different fractions exhibited good DPPH radical and ABTS radical cation scavenging activities and strong inhibitory effects on the generation of intracellular reactive oxygen species (ROS). Moreover, these three fractions also had good inhibitory effects towards α-glucosidase and pancreatic lipase. Among the three fractions, the F fraction possessed the strongest bioactivities. The findings obtained in the current study may provide some insights and bases for the further investigation and application of G. biloba nut shells in clinical medicine or the nutraceutical industry.

Simultaneous determination of ginkgolide A, B, C, bilobalide and rutin in rat plasma by LC-MS/MS and its application to a pharmacokinetic study

Abstract A reliable LC-MS/MS method for the determination of five bioactive constituents (bilobalide, BLL; ginkgolide A, GLA; ginkgolide B, GLB; ginkgolide C, GLC; rutin) of Ginkgo biloba leaf extracts (GBE) in rat plasma was established, fully validated and applied to an intragastric pharmacokinetic study of a preparation of GBE in rat. Samples were extracted with ethyl acetate. C18 column was selected as analytical column in this method. Mobile phase was water with 0.01% formic acid and acetonitrile. Quantification was performed in negative multiple-reaction monitoring mode. Matrix instability of terpene lactones was noticed and hydrochloric acid was used as a stabilizer. This method showed good precision and accuracy, recovery was reproducible and matrix effect was negligible. Among four terpene lactones, BLL had the highest exposure and the shortest terminal half-life, GLA and GLB had lower exposure and longer terminal half-life, the exposure of GLC was lowest and its terminal half-life was the maximum, and all of them showed rapid absorption. This study provides a reference for determination of terpene lactones and flavonol glycoside prototypes in GBE and offers pharmacokinetic data of flavonol glycoside prototype in GBE.

Therapeutic Versus Preventative Use of Ginkgo biloba Extract (EGb 761) against Indomethacin-Induced Gastric Ulcer in Mice

The main bioactive constituents in the standardized Ginkgo biloba leaf extract (EGb 761) are the terpene lactones and flavonoid glycosides. EGb 761’s antioxidant and anti-inflammatory properties have previously been demonstrated. Indomethacin-induced gastric ulcers have a multifactorial etiology and represent a major restriction to its therapeutic utility. The underlying ulcerogenic process involves oxidative and inflammatory biomolecular insults. This study was performed to explore the curative and preventative benefits of EGb 761 in experimentally-induced ulcers. To develop gastric ulcers in mice, indomethacin (40 mg/kg) was administered orally. EGb 761 (200 mg/kg) was given by gavage for 7 days before (preventative) and after (therapeutic) indomethacin administration. The histological alterations and macroscopic mucosal lesions were assessed. In gastric tissue homogenates, malondialdehyde (MDA), reduced glutathione (GSH), nitric oxide (NO), and inflammatory cytokines were measured. The expressions of cyclooxygenase-2 (COX-2), cytokines, and proliferating cell nuclear antigen (PCNA) in the stomach mucosa were also investigated. The ulcer index, histological alterations, gastric oxidants, and inflammatory biomarkers were all significantly increased by indomethacin. In stomach specimens, it increased COX-2 and PCNA expression. EGb 761 treatments, both prophylactic and therapeutic, resulted in significant reductions in ulcer lesions, nitrosative and oxidative damage, and inflammatory markers, along with the lowering of COX-2 and PCNA expressions. Furthermore, in the fight against stomach ulcers, EGb 761 treatment was found to be more efficient than prevention.

Effect of origin, tree age, and harvesting time on content of flavonoids and terpene lactones in Ginkgo Folium

The content of total flavonol glycosides in Ginkgo Folium in the planting bases was determined by high performance liquid chromatography(HPLC).The samples were extracted by reflux with methanol-25% hydrochloric acid.The HPLC conditions were as follows: Agilent ZORBAX SB-C_(18) column(4.6 mm×250 mm, 5 μm), isocratic elution with mobile phase of 0.4% phosphoric acid solution-methanol(45∶55), flow rate of 1 mL·min~(-1), column temperature of 30 ℃, detection wavelength of 360 nm, and injection vo-lume of 10 μL.A method for the determination of terpene lactones in Ginkgo Folium was established based on ultra-high performance liquid chromatograph-triple-quadrupole/linear ion-trap tandem mass spectrometry(UPLC-QTRAP-MS/MS).The UPLC conditions were as below: gradient elution with acetonitrile-0.1% formic acid, flow rate of 0.2 mL·min~(-1), column temperature of 30 ℃, sample chamber temperature of 10 ℃, and injection volume of 10 μL.The ESI~+and multiple reaction monitoring(MRM) were adopted for the MS.The above methods were used to determine the content of total flavonol glycosides and terpene lactones in 99 batches of Ginkgo Folium from 6 planting bases, and the results were statistically analyzed.The content of flavonoids and terpene lactones in Ginkgo Folium from different origins, from trees of different ages, harvested at different time, from trees of different genders, and processed with different methods was compared.The results showed that the content of total flavonol glucosides in 99 Ginkgo Folium samples ranged from 0.38% to 2.08%, and the total content of the four terpene lactones was in the range of 0.03%-0.87%.The method established in this study is simple and reliable, which can be used for the quantitative analysis of Ginkgo Folium.The research results lay a basis for the quality control of Ginkgo Folium.

The qualitative and quantitative profiling for quality assessment of Yinxing Mihuan Oral Solution and the stability study on the focused flavonol glycosides

Yinxing Mihuan Oral Solution (YMOS) has been widely applied for the treatment of coronary heart disease, angina pectoris, and cerebral ischemic disease in clinical practice. Nonetheless, the limited basic researches on quality analysis of YMOS remain a critical bottleneck that needs to be enhanced for better clinical applications. In this study, a total of 67 chemical components, including flavonoids, terpene lactones, nucleosides, etc., were tentatively characterized by ultra-high performance liquid chromatography tandem Q-Exactive Orbitrap high-resolution mass spectrometry, among which 34 compounds were further identified by comparison with reference substances. By adopting a methodologically validated method, we discovered that the quantitative estimate of multi-compounds in 22 batches of YMOS showed lot-to-lot consistency, and the additives in YMOS also met the corresponding regulations. Furthermore, five flavonol glycosides whose content presented a downward trend in the expired YMOS were focused. A systematic research on stability test focusing on the five targeted flavonol glycosides was performed under different temperatures and pH levels. It was found that ortho-diphenolic hydroxyl group on B-ring and the type of saccharide connected to 3-hydroxyl on C-ring play a pivotal role in the stability of the tested compounds. Subsequently, as the important compounds, ginkgolides A, B, and C in YMOS were simultaneously quantified with ultra performance liquid chromatography coupled with triple quadrupole mass spectrometry. In brief, this study performs a reliable chemical identification and provides a rapid and feasible method for the quality evaluation, which contributes to the in-depth investigation and safe application of YMOS for clinical uses.

Cardiovascular risk factors for cognitive deficiency and options for correction

Cardiovascular risk factors for the development of cognitive impairment and Alzheimer’s disease are ranked among the most significant ones, especially in elderly patients. The review provides data on the influence of such factors as arterial hypertension, arterial hypotension, heart failure, coronary heart disease, etc. on the development of cognitive deficiency. The presence of two or more of these risk factors generally tends to increase the possibility of the development of Alzheimer’s disease. The author examines the relationship between these factors and the development of Alzheimer’s disease. Insights into how CVD risk factors can cause progressive cognitive impairment provides further insight into the pathophysiology of Alzheimer’s disease and the identification of methods to prevent or treat CV risk factors for cognitive decline. The course and outcome of cardiovascular diseases depend on the patient’s age, previous medical history, lifestyle, primary prevention, genetic and pathological factors that affect structural and hemodynamic functions. The article addresses issues of therapy of these disorders. Methods of non-pharmacological therapy are discussed: the importance of the patient’s lifestyle, physical activity, nutritional issues, social activity of patients. Much attention is paid to the role of cognitive training in the management of patients with cognitive impairments. The correction of risk factors for stroke and the progression of chronic circulatory disorders is one of the most elaborated areas. The therapeutic potential drugs with a multifactorial mechanism of action are discussed. One of them is Ginkgo biloba extract EGb 761®, which meets the regulated content of active ingredients (flavone glycosides, terpene lactones and ginkgolic acid) and, perhaps, promises to be a more rational choice in the treatment of cognitive impairment. The role of Ginkgo biloba extract EGb 761® in the treatment of cardiovascular diseases is discussed.

Ginkgo Biloba: A Famous Living Fossil Tree and an Ancient Herbal Traditional Chinese Medicine

Background: Ginkgo (Ginkgo biloba) is a living fossil and a deciduous tree, having extracts with antidiabetic, antioxidant, anticancer, antihypertensive, immunostimulating, hepatoprotective, and antimicrobial activities, memory enhancement efficiency, and beneficial effects against neurodegenerative disease. Objective: This study aimed at evaluating the medicinal values and natural benefits of Ginkgo. Methods: This review investigated publication in MEDLINE/PubMed database and Google Scholar. The keywords used for an electronic search were Ginkgo biloba, living fossil, bioactive components, and traditional Chinese medicine. Results: The main active constituents of Ginkgo biloba are flavone glycosides such as kaempferol, quercetin, and isorhamnetin, terpene lactones, alkylphenols, proanthocyanidins, rhamnose, glucose, D-glucaric acid, ginkgolic acid, organic acids such as hydroxykinurenic, kynurenic, protocatechic, shikimic, and vanillic. Ginkgo kernels have been used as medicine or eaten as nuts in traditional medicinal science. The most notable pharmaceutical applications of Ginkgo are observed in cardiovascular disease, Alzheimer's disease, impaired cerebral performance, vascular insufficiency, antidepressant-induced sexual dysfunction, premenstrual syndrome, liver fibrosis, vascular disease, tinnitus, macular degeneration, memory, and vertigo. Conclusion: The development of modern drugs from Ginkgo by considering the importance of traditional medicinal Asian science with further research works should be emphasized.

Application of near infrared spectroscopy to predict contents of various lactones in chromatographic process of Ginkgo Folium

This study established a method for rapid quantification of terpene lactone, bilobalide, ginkgolide C, ginkgolide A and ginkgolide B in the chromatographic process of Ginkgo Folium based on near infrared spectroscopy(NIRS). The effects of competitive adaptive reweighting sampling(CARS), random frog(RF), and synergy interval partial least squares(siPLS) on the performance of partial least squares regression(PLSR) model were compared to the reference values measured by HPLC. Among them, the correlation coefficients of prediction(Rp) of validation sets of terpene lactone, bilobalide, and ginkgolide C were all higher than 0.98, and the relative standard errors of prediction(RSEPs) were 5.87%, 6.90% and 6.63%, respectively. Aiming at ginkgolide A and ginkgolide B with relatively low content, the genetic algorithm joint extreme learning machine(GA-ELM) was used to establish the optimized quantitative analysis model. Compared with CARS-PLSR model, the CARS-GA-ELM models of ginkgolide A and ginkgolide B exhibited a reduction in RSEP from 15.65% to 8.52% and from 21.28% to 10.84%, respectively, which met the needs of quantitative ana-lysis. It has been proved that NIRS can be used for the rapid detection of various lactone components in the chromatographic process of Ginkgo Folium.

Optimization of terpene lactones and ginkgolic acids extraction from Ginkgo biloba L. leaves by natural deep eutectic solvents using experimental design and HPTLC-MS analysis

In this work, a green extraction method was investigated to maximize the yield of terpene lactones and minimize that of ginkgolic acids from Ginkgo biloba L. leaves.An HPTLC-MS method for identification of simultaneous determination of terpene trilactones and ginkgolic acids in G. biloba samples was developed and validated. The limit of detection is 0.791, 0.850, 0.868, 0.785, 0.763, 0.871 and 0.622 μg/band, whereas the limit of quantitation is 2.399, 2.576, 2.632, 2.380, 2.313, 2.640 and 1.885 μg/band for bilobalide, ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide K, ginkgolide J and ginkgolic acid, correspondingly. The extraction ability of 15 different mixtures of NADES, using ultrasonic assisted extraction in comparison to methanol, ethanol and water was tested. The results revealed that DES2 (Choline chloride, ascorbic acid) displayed the highest extraction yields of total ginkgolides. Concerning ginkgolic acids, in comparison with conventional solvents, DES 2 displayed significantly lower extraction amounts. Optimization experiments using Box Behnken design indicated that the ginkgolides extraction yield was highly affected by water percentage (C, p < 0.0001; C2, p < 0.0001) followed by ultrasonication temperature (B, p < 0.0001; B2, p < 0.0001), while ultrasonication temperature possessed stronger influence on extraction yield of ginkgolic acids. The set optimum conditions for extraction were found to be an ultrasonication time of 36.63 min, temperature of 55.77 °C and a water content of 33.66% v/v.

JUSTIFICATION OF THE FEASIBILITY OF CREATING NEW GINKGO BILOBA MEDICINES AND DIETARY SUPPLEMENTS BASED ON THE STUDY OF THE UKRAINIAN PHARMACEUTICAL MARKET

Formulation of the problem. In view of the socio-economic crisis in Ukraine, the process of introducing new medicines to the market, ensuring their availability for all segments of the population remains a rather important problem in the country’s health care system in today’s conditions. Recently, medicinal products and dietary supplements based on ginkgo biloba have been widely used in modern pharmacotherapy. This is due to the unique phytochemical composition, which includes a number of powerful antioxidants and other active substances. Analysis of recent research and publications. Comprehensive works on marketing research of the domestic pharmaceutical market on the example of medicines and dietary supplements based on dry Ginkgo biloba leaf extract in the directions outlined in this work have not been conducted. Today, the attention of scientists is largely focused on the use in medical practice and dietetics of products based on Ginkgo biloba leaf extract due to their natural origin and multi-vector range of pharmacotherapy. Despite the interest of scientists in the pharmacological properties of the extract from the leaves of Ginkgo biloba, there is almost no information about the feasibility of finding and developing new herbal remedies based on it. The above determined the expediency of the work, determined its purpose, tasks, structure and logical construction of the research sequence. Formulating the purpose of the article. The purpose of the work was to conduct marketing research of the domestic pharmaceutical market in the segment of drugs and dietary supplements based on dry Ginkgo biloba leaf extract to determine marketing opportunities for domestic practical pharmacy. The presentation of the main material. Ginkgo biloba is a natural source of biologically active compounds. A significant amount of terpene lactones and flavonglycosides in the composition of natural raw materials, which have powerful antioxidant and anti-inflammatory properties and have a beneficial effect on the most important systems of the human body – the brain and blood circulation, as well as contribute to the normalization of metabolism, prevent premature aging of tissues, etc. The assortment of medicines and dietary supplements based on the extract of dry Ginkgo biloba leaves in the retail segment of the pharmaceutical market of Ukraine was analyzed. The main countries that export to the Ukrainian pharmaceutical market medicinal products and dietary supplements based on dry Ginkgo biloba leaf extract are the USA, Slovenia and Germany. Medicinal products and dietary supplements based on extract from dry Ginkgo biloba leaves are mostly presented on the domestic market in the form of capsules and tablets. The socio-economic availability of medicines and dietary supplements based on dry Ginkgo biloba leaf extract was determined based on the calculation of marketing indicators: the price liquidity ratio and the solvency adequacy ratio. The advantages and disadvantages of using certain extractants for obtaining dry Ginkgo biloba extract are given and substantiated. Given the resource efficiency, simplicity and reliability of the equipment used, the use of ethyl alcohol as an extractant is proposed. A brief description of the technological stages of production of Ginkgo biloba dry extract is proposed and given. Conclusions and prospects for further research. Providing all sections of the country’s population with affordable, effective and safe medicines is one of the priority and strategically important tasks of today’s pharmacy. The conducted marketing research of the pharmaceutical market in the segment of medicines and dietary supplements based on the extract of dry Ginkgo biloba leaves can be considered as a basis for the search and development of new effective phytoremedies of domestic production for the treatment of socially dangerous diseases.

Identification of Chemical Components of Qi-Fu-Yin and Its Prototype Components and Metabolites in Rat Plasma and Cerebrospinal Fluid via UPLC-Q-TOF-MS

Qi-Fu-Yin, a traditional Chinese medicine formula, has been used to treat Alzheimer's disease (AD, a neurodegenerative disorder) in clinical setting. In this study, the chemical components of Qi-Fu-Yin and its prototype components and metabolites in rat plasma and cerebrospinal fluid, after oral administration, were preliminarily characterized via ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS). A total of 180 compounds, including saponins, flavonoids, organic acids, sucrose esters, oligosaccharide esters, phthalides, phenylethanoid glycosides, alkaloids, xanthones, terpene lactones, ionones, and iridoid glycoside, were tentatively characterized. For the first time, 51 prototypical components and 26 metabolites, including saponins, phthalides, flavonoids, sucrose esters, organic acids, alkaloids, ionones, terpene lactones, iridoid glycoside, and their derivatives, have been tentatively identified in the plasma. Furthermore, 10 prototypical components (including butylidenephthalide, butylphthalide, 20(S)-ginsenoside Rh1, 20(R)-ginsenoside Rh1, and zingibroside R1) and 6 metabolites were preliminarily characterized in cerebrospinal fluid. These results were beneficial to the discovery of the active components of Qi-Fu-Yin anti-AD.

Genome-wide characterization of bZIP gene family identifies potential members involved in flavonoids biosynthesis in Ginkgo biloba L.

Ginkgo biloba L. is an ancient relict plant with rich pharmacological activity and nutritional value, and its main physiologically active components are flavonoids and terpene lactones. The bZIP gene family is one of the largest gene families in plants and regulates many processes including pathogen defense, secondary metabolism, stress response, seed maturation, and flower development. In this study, genome-wide distribution of the bZIP transcription factors was screened from G. biloba database in silico analysis. A total of 40 bZIP genes were identified in G. biloba and were divided into 10 subclasses. GbbZIP members in the same group share a similar gene structure, number of introns and exons, and motif distribution. Analysis of tissue expression pattern based on transcriptome indicated that GbbZIP08 and GbbZIP15 were most highly expressed in mature leaf. And the expression level of GbbZIP13 was high in all eight tissues. Correlation analysis and phylogenetic tree analysis suggested that GbbZIP08 and GbbZIP15 might be involved in the flavonoid biosynthesis. The transcriptional levels of 20 GbbZIP genes after SA, MeJA, and low temperature treatment were analyzed by qRT-PCR. The expression level of GbbZIP08 was significantly upregulated under 4°C. Protein–protein interaction network analysis indicated that GbbZIP09 might participate in seed germination by interacting with GbbZIP32. Based on transcriptome and degradome data, we found that 32 out of 117 miRNAs were annotated to 17 miRNA families. The results of this study may provide a theoretical foundation for the functional validation of GbbZIP genes in the future.

Chemical Constituents Isolated from the Leaves of Toricellia angulata Oliv. var. intermedia (Harms.) Hu

In our survey on the chemical composition of Chinese folk medicines, nine compounds were isolated from methanol extract of the leaves of Toricellia angulata Oliv. var. intermedia (Harms.) Hu (Corniaceae). The structures of these compounds were elucidated on the basis of NMR data analysis, which were identified as dimethyl 2-(hydroxymethyl)-5-oxocyclohexane-1,4-dicarboxylate (1), methyl succinate (2), 5-hydroxymethyl-2-furfuraldehyde (3), 7-hydroxy-6-methoxycoumarin (4), loliolide (5), (8 S)-deca-2-trans-2,9-diene-4,6-diyn-1,8-diol (6), methyl malate (7), griselinoside (8), and methyl linoleate (9), respectively. Among them, compound 1 is a new cyclohexanone derivative and given a trivial name torriangulate A, while others are categorized to be organic acids (2, 7, and 9), a coumarin derivative (4), a terpene lactone (5), a polyacetylene (6), and an iridoid glycoside (8). Compounds 3–5 were isolated from this genus for the first time and compound 9 was first identified from this species. The discovered compounds with novel or known structures further reveal the chemical basis of T. angulata var . intermedia, which lays a foundation for the development of T. angulata var . intermedia used as a traditionally folk medicine.

Integration of morphological, physiological and multi-omics analysis reveals the optimal planting density improving leaf yield and active compound accumulation in Ginkgo biloba

The leaves of Ginkgo biloba contain abundant active compounds, including flavonoids and terpene lactones. Leaves of 1–5-year-old seedlings are typically harvested for ginkgo leaf extract, which is used for treatment of cardiovascular diseases, dementia, and dizziness. However, the effects of planting densities on leaf yield and leaf active compound accumulation in ginkgo are largely unknown. Here, we conducted field experiments at four planting densities (control, moderate density [MD], medium-high density [MHD] and high density [HD]) of ginkgo seedlings to investigate their leaf production and quality. As planting density increased, stem elongation was enhanced whereas root growth was inhibited. In addition, soluble sugar and starch contents were decreased in leaves with density increasing. Although the fresh and dry weights of leaves per plant were reduced, the leaf yield per unit area increased by 60.5–138.8 % in two successive years at MD-HD. Interestingly, compared with other densities, MD resulted in a higher accumulation of various bioactive secondary metabolites, including 13 flavonoids (such as homoeriodictyol, baimaside, and ladanein), ginkgolides, and bilobalide. Correspondingly, the genes encoding chalcone synthase (CHS), chalcone isomerase (CHI), and flavonol synthase (FLS), which are involved in flavonoid biosynthesis, were upregulated. However, when exceeding MD, the contents of bioactive secondary metabolites and the expression of related genes decreased dramatically. These results suggest that the MD as the optimal density could not only enhance leaf yield, but also improve active-compound accumulation in G. biloba seedling leaves, providing a practical and theoretical reference for leaf-use ginkgo plantation.

Draft genome sequencing and functional annotation and characterization of biofilm-producing bacterium Bacillus novalis PD1 isolated from rhizospheric soil.

Biofilm forming bacterium Bacillus novalis PD1 was isolated from the rhizospheric soil of a paddy field. B. novalis PD1 is a Gram-positive, facultatively anaerobic, motile, slightly curved, round-ended, and spore-forming bacteria. The isolate B. novalisPD1 shares 98.45% similarity withB. novalis KB27B. B. vireti LMG21834 and B. drentensis NBRC 102,427 are the closest phylogenetic neighbours for B. novalis PD1. The draft genome RAST annotation showed a linear chromosome with 4,569,088bp, encoding 6139 coding sequences, 70 transfer RNA (tRNA), and 11 ribosomal RNA (rRNA) genes. The genomic annotation of biofilm forming B. novalis PD1(> 3.6@OD595nm) showed the presence of exopolysaccharide-forming genes (ALG, PSL, and PEL) as well as other biofilm-related genes (comER, Spo0A, codY, sinR, TasA, sipW, degS, and degU). Antibiotic inactivation gene clusters (ANT (6)-I, APH (3')-I, CatA15/A16 family), efflux pumps conferring antibiotic resistance genes (BceA, BceB, MdtABC-OMF, MdtABC-TolC, and MexCD-OprJ), and secondary metabolites linked to phenazine, terpene, and beta lactone gene clusters are part of the genome.