Full-length transcriptome characterization and comparative analysis of Gleditsia sinensis.

As an economically important tree, Gleditsia sinensis Lam. is widely planted. A lack of background genetic information on G. sinensis hinders molecular breeding. Based on PacBio single-molecule real-time (SMRT) sequencing and analysis of G. sinensis, a total of 95,183 non-redundant transcript sequences were obtained, of which 93,668 contained complete open reading frames (ORFs), 2,858 were long non-coding RNAs (LncRNAs) and 18,855 alternative splicing (AS) events were identified. Genes orthologous to different Gleditsia species pairs were identified, stress-related genes had been positively selected during the evolution. AGA, AGG, and CCA were identified as the universal optimal codon in the genus of Gleditsia. EIF5A was selected as a suitable fluorescent quantitative reference gene. 315 Cytochrome P450 monooxygenases (CYP450s) and 147 uridine diphosphate (UDP)-glycosyltransferases (UGTs) were recognized through the PacBio SMRT transcriptome. Randomized selection of GsIAA14 for cloning verified the reliability of the PacBio SMRT transcriptome assembly sequence. In conclusion, the research data lay the foundation for further analysis of the evolutionary mechanism and molecular breeding of Gleditsia.

Application of choline chloride deep eutectic solvents and high-speed counter-current chromatography to the extraction and purification of flavonoids from the thorns of Gleditsia sinensis Lam.

INTRODUCTION: Flavonoids are the most important and effective constituents in the thorns of Gleditsia sinensis Lam., which have been known to show antimicrobial, antiviral, anticancer, and anticoagulant activities. However, efficient extraction and separation methods for these flavonoids are not currently established. OBJECTIVE: To develop an efficient method for efficient extraction and rapid separation of flavonoids from the thorns of G. sinensis using choline chloride deep eutectic solvents (DESs) and high-speed counter-current chromatography (HSCCC). METHODOLOGY: As for extraction, DES composed of choline chloride and 1,4-butanediol at 1:4 mole ratio, at an extraction temperature of 55°C, 20% of water content, 1:30 mg/mL for solid-liquid ratio, and 45 min for extraction time were selected as the optimised extraction method for flavonoids from the thorns of G. sinensis. As for separation, dichloromethane-methanol-n-butanol-water (4:3:0.5:2, v/v) was applied to develop a successful strategy for purification of the flavonoids by HSCCC. RESULTS: Totally, five flavonoids, including padmatin (1, 3.7 mg), isovitexin (2, 2.5 mg), 3',5,5',7-tetrahydroxyflavanonol (3, 11.2 mg), 7,4'-dihydroxy-5,3'-dimethoxyflavanonol (4, 4.1 mg), and quercetin (5, 3.8 mg), were successfully obtained from 250 mg of the extracted flavonoids by HSCCC. CONCLUSION: Results demonstrated that the combination of DES and HSCCC is a powerful technique for the extraction, and isolation of flavonoids from the thorns of G. sinensis compared with conventional organic solvent extraction and column chromatography, which have been proven to provide higher extraction efficiency for flavonoids and rapidly obtain the quality control markers of flavonoids from the investigated plant.

Effects of Gleditsia sinensis pod powder, coconut shell biochar and rice husk biochar as additives on bacterial communities and compost quality during vermicomposting of pig manure and wheat straw.

This study investigated the effectiveness of Gleditsia sinensis pod powder (GSPP), coconut shell biochar (CSB), rice husk biochar (RHB) and their mixtures on vermicomposting of pig manure and wheat straw using Eisenia fetida. The results indicated that the addition of GSPP or/and CSB and RHB could greatly enhance the relative abundance of Bacteroidetes, Actinobacteria, and Firmicutes, as well as the activities of celluloses, protease, and alkaline phosphatase. However, the earthworm biomass was increased in the GSPP and/or CSB addition treatments but decreased in RHB addition treatments compared with the control. Compared with the control, addition of 4%GSPP+8%CSB significantly (P < 0.05) accelerated the degradation of organic matter and increased the concentration of nutrients (total N, P, K), NO3--N in final vermicompost. Germination and growth of tomato seedings were also higher (P < 0.05) in vermicompost produced with the addition of 4%GSPP+8%CSB than in control. Consequently, 4%GSPP+8%CSB addition was suggested as an efficient method to improve the vermicomposting of pig manure and wheat straw.

Genome-wide analysis of microsatellite and sex-linked marker identification in Gleditsia sinensis.

BACKGROUND: Gleditsia sinensis Lam. (Leguminosae), a dioecious perennial arbor, demonstrates important medicinal properties and economic value. These properties can be harnessed depending on the sex of the plant. However, the sex of the plants is difficult to identify accurately through morphological methods before the flowering. RESULTS: We used bulked segregant analysis to screen sex-specific simple sequence repeat (SSR) markers in G. sinensis. Five male and five female plants were pooled to form the male and female bulks, respectively, and subjected to whole-genome sequencing. After high-throughput sequencing, 5,350,359 sequences were obtained, in which 2,065,210 SSRs were searched. Among them, the number of duplicated SSRs was the highest. The male plants could reach 857,874, which accounted for 60.86% of the total number of male plants. The female plants could reach 1,447,603, which accounted for 56.25% of the total model of the female plants. Among all the nucleotide repeat types, the A/T-rich motif was the most abundant. A total of 309,516 female strain-specific SSRs were selected by clustering. After designing the primers, the male and female gene pools were amplified, and five pairs of primers (i.e., 27, 34, 36, 39, and 41) were found to amplify the differential bands in the male and female gene pools. Using the five pairs of primers, we performed PCR verification on 10 individuals of known sex, which constructed the gene pool. The female plants amplified a single fragment of lengths (i.e., 186, 305, 266, 203, and 260 bp) and no male plant strip, thereby completing the identification of the male and female sexes of the G. sinensis. CONCLUSIONS: This study provides accurate sex identification strategies between female and male plants, thus improving the utilization rate of G. sinensis resources.

Chemical constituents from the thorns of Gleditsia sinensis and their cytotoxic activities.

A new aromatic glycoside (1) and a new natural product, neolignan (2), along with twenty-three known compounds (3-25), were isolated from the thorns of Gleditsia sinensis. According to the spectroscopic analyses (IR, UV, HRESIMS, NMR and ECD), the structures of isolates were elucidated. Herein, compounds 4, 6-8, 10-13, 15, 16, 18, 20, 23 were isolated from the plant of G. sinensis for the first time. Moreover, compounds 4, 6, 15 and 24 showed cytotoxic effects on human ovarian cancer (SKOV-3) cells with IC50 values of 24.83 ± 4.90, 48.86 ± 9.11, 80.13 ± 5.62, 15.38 ± 2.21 µM, respectively. [Formula: see text].

Valorization of Gleditsia triacanthos Invasive Plant Cellulose Microfibers and Phenolic Compounds for Obtaining Multi-Functional Wound Dressings with Antimicrobial and Antioxidant Properties.

Gleditsia triacanthos is an aggressive invasive species in Eastern Europe, producing a significant number of pods that could represent an inexhaustible resource of raw material for various applications. The aim of this study was to extract cellulose from the Gleditsia triacanthos pods, characterize it by spectrophotometric and UHPLC-DAD-ESI/MS analysis, and use it to fabricate a wound dressing that is multi-functionalized with phenolic compounds extracted from the leaves of the same species. The obtained cellulose microfibers (CM) were functionalized, lyophilized, and characterized by ATR-FTIR and SEM. The water absorption and retention capacity as well as the controlled release of phenolic compounds with antioxidant properties evaluated in temporal dynamics were also determined. The antimicrobial activity against reference and clinical multi-drug-resistant Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterobacter cloacae, Candida albicans, and Candida parapsilosis strains occurred immediately after the contact with the tested materials and was maintained for 24 h for all tested microbial strains. In conclusion, the multi-functionalized cellulose microfibers (MFCM) obtained from the reproductive organs of an invasive species can represent a promising alternative for the development of functional wound dressings with antioxidant and antimicrobial activity, as well as being a scalable example for designing cost-effective, circular bio-economy approaches to combat the accelerated spread of invasive species.

Seed local adaptation and seedling plasticity account for Gleditsia triacanthos tree invasion across biomes.

BACKGROUND AND AIMS: Phenotypic plasticity and local adaption can contribute to the success of invasive species. While the former is an environmentally induced trait, the latter involves a selection process to filter the best genotype for a location. We examined the evidence for phenotypic plasticity and local adaptation for seed and seedling traits of the invasive tree Gleditsia triacanthos, with three origins distributed along an approx. 10° latitude gradient across three biomes. METHODS: In sub-tropical forests, dry woodlands and secondary temperate grasslands in Argentina, we harvested seeds from clusters of neighbouring trees (i.e. families) distributed within 15-20 km in each origin (biome). We manipulated the environmental conditions relevant to each biome, assuming that propagule availability did not represent an ecological barrier. In growth chambers, we evaluated seed imbibition and seed germination under different light, temperature and water potential. In a 2 year common garden, we evaluated the impact of resident vegetation removal on seedling survival and growth. KEY RESULTS: Mean time to complete seed imbibition differed among origins; seeds from temperate grasslands reached full imbibition before seeds from dry woodlands and sub-tropical forests. Germination was always >70 %, but was differentially affected by water potential, and light quantity (dark-light) and quality (red-far red) among origins, suggesting local adaptation. In the common garden, vegetation removal rather than origin negatively affected seedling survival and enhanced seedling growth. Vegetation removal increased basal diameter, leaves per plant and spine number, and reduced the height:basal diameter ratio. CONCLUSIONS: We conclude that local adaptation in seed germination traits and plastic changes in seedling allometry (e.g. height:diameter) may allow this tree to respond over the short and long term to changes in environmental conditions, and to contribute to shape G. triacanthos as a successful woody invader. Overall, our study revealed how local adaptation and plasticity can explain different aspects of tree invasion capacity across biomes.

Control of biofouling on pearl oysters Pinctada imbricata using wax and Chinese herbs.

Biofouling poses severe challenges to pearl oyster Pinctada imbricata culture in China, and controlling it is both labor- and capital-intensive. The antifouling properties of wax, and wax mixed with Chinese herbs, sprayed onto pearl oyster shell surfaces during peak biofouling seasons were evaluated. Pearl oysters coated with three wax treatments (plain wax, Chinaberry seed extract, Chinese honeylocust fruit extract) and a control (no treatment), were cultured in nets for up to 60 days. Mortality rate, fouling organism and pearl-oyster weights, and shell height are reported for individual oysters on each of six sampling dates. With the exception of oysters submerged for 12 days, all oysters were significantly affected by treatment type and submersion duration. Fouling weight increased more rapidly over time in the control-treatment oysters. Wax-based coatings deterred fouling-organism settlement on oysters for at least 2 months during the intensive fouling season, reducing mortality and not adversely effecting growth.

Can arbuscular mycorrhizal fungi from non-invaded montane ecosystems facilitate the growth of alien trees?

It is generally assumed that recruitment and expansion of alien species along elevation gradients are constrained by climate. But, if plants are not fully constrained by climate, their expansion could be facilitated or hindered by other factors such as biotic interactions. Here, we assessed the composition of arbuscular mycorrhizal fungi (AMF) in soils along an elevation gradient (i.e. 900 m, 1600 m, 2200 m and 2700 m a.s.l.) through a fungal DNA meta-barcoding approach. In addition, we studied in the greenhouse the effects of AMF on growth and phosphorous (P) nutrition of seedlings of the alien trees Gleditsia triacanthos, Ligustrum lucidum and Pyracantha angustifolia cultivated in soils from those elevations, spanning the elevation at which they already form monospecific stands (below 1450 m a.s.l.) and higher elevations, above their current range of distribution in montane ecosystems of Central Argentina. For comparison, we also included in the experiment the dominant native tree Lithraea molleoides that historically occurs below 1300 m a.s.l. Arbuscular mycorrhizal fungal community composition showed strong community turnover with increasing elevation. The effects of these AMF communities on plant growth and nutrition differed among native and alien trees. While P nutrition in alien species' seedlings was generally enhanced by AMF along the whole gradient, the native species benefited only from AMF that occur in soils from the elevation corresponding to its current altitudinal range of distribution. These results suggest that AMF might foster upper range expansion of these invasive trees over non-invaded higher elevations.

HPLC-MS/MS determination of flavonoids in Gleditsiae Spina for its quality assessment.

Gleditsiae Spina, the thorn of Gleditsia sinensis Lam., has been used as an anti-inflammatory, anti-tumor, and anti-bacterial traditional medicine for hundreds of years in China. This study used high-performance liquid chromatography and tandem mass spectrometry combined with chemometric methods to allow the fast and accurate identification and quantification of the flavonoids compounds in Gleditsiae Spina, and created reliable criteria for accurate identification of Gleditsiae Spina and its adulterants. This research provides good evidence for the classification and quality evaluation of Gleditsiae Spina. Firstly, eight flavonoids compounds were detected and identified on the basis of their mass spectra, fragment characteristics, and comparison with published data. Then the mass spectroscopic fragmentation pathways of these compounds were determined and, in addition rutin, isoquercitrin, and quercitrin were detected in Gleditsiae Spina for the first time. The quantification was performed on a triple quadrupole tandem mass spectrometer in multi-reaction monitoring mode, and the baseline separation of the eight bioactive flavonoids components was achieved within 13 min. Furthermore, the proposed method was successfully applied for simultaneous quantitative determination of the eight Gleditsiae Spina compounds and adulterants obtained from different sources in China. Then, we built a classification model which showed a high level of accuracy predicting 100% of the samples, correctly.

Sulfated Purine Alkaloid Glycosides from the Pupal Case Built by the Bruchid Beetle Bruchidius dorsalis Inside the Seed of Gleditsia japonica.

Three new sulfated isoguanine alkaloid glycosides, designated as saikachinoside A monosulfate (1), saikachinoside A disulfate (2), and locustoside B disulfate (3), have been isolated from the pupal case of the wild bruchid seed beetle Bruchidius dorsalis (Chrysomelidae, Bruchinae) infesting the seed of Gleditsia japonica Miq. (Fabaceae). Their structures were determined by spectroscopic methods and the inhibitory activity of 2 and 3 against acid phosphatase was evaluated.

[Research on morphological characteristics change of quercetin and polyphenols of accumulation dynamic of spines of Gleditsia sinensis].

To study morphological characteristics change of the development process and quercetin and polyphenols of accumulation dynamic of the spines of Gleditsia sinensis, measure and compare morphological indexes using Vernier caliper, ruler and balance, calibrate and analysis quercetin and polyphenols content using HPLC and colorimetric method. The spines of G. sinensis development process is divided into formation period (the beginning of August to the beginning of November), dormant period (the beginning of November to the end of March in the following year), germination period (the end of March to the middle of April), fast growth period (the middle of April to the middle of August), browning period (the middle of August to the end of August) and mature period (the beginning of September to the end of December). Formation period the spines primordium divides and forms the scale bubs; dormant period the scale bubs are in a dormant state; germination period the bubs scales fall off, spines primordium began to development; fast growth period rapid growth to maximum; browning period browning from the tip and browns until the whole becomes brown; mature period The early stage of maturity is full of luster, gradually the color deepened and the luster faded. The accumulation of quercetin was gradually decreasing after increasing. The total polyphenol accumulation was significantly decreased and then gradually increased, decreased finally. The content of quercetin was increased from 0.000 4%-0.002 6%, and the polyphenol content decreased from 0.761 9%-0.049 1% and then slowly increased to 0.286 9% in the fast growth period.The quercetin continuous increase to 0.004 3% and total polyphenol increased to 0.421 6% in the browning period. In the mature period, the quercetin content significantly decreased after reaching 0.009 6% in September, and the polyphenols content decreased after reaching 0.723 5% in October. Using principal component analysis results: September first, October 2nd, November 3rd. The morphological characteristics change of the development process and quercetin and polyphenols accumulation were determined. The development process is divided into six periods, the best harvest time is the early stage of mature period. Provide theoretical support for the utilization of the spines of G. sinensis and cultivation techniques of high yield.

[Study on breeding and pollination characteristics of Gleditsia sinensis].

To study the breeding system and pollination characteristics of Gleditsia sinensis, we observed the development of flower development and the processing of pollination, and determined the pollen viability and stigma acceptability by TTC and benzidine-hydrogen peroxide method and detected its breeding system using OCI value, P/O ratio and artificial pollination.The results showed that: ①G. sinensis are racemes, divided into bisexual inflorescences (only a small amount of inactive pollen) and male inflorescences (occasionally a few bisexual flowers), flowers hermaphrodite. ②Male flowers had the strongest pollen activity 4 h after flowering; the stigma receptivity of bisexual was the highest at 1 h after flowering, and pollination was the best in this time. ③The pollen tube had a few elongation when the bisexual flower is half-opened. The number of pollen tube and length significantly increased when blooming. The flower reaches the ovary and even enters the ovule to complete the fertilization. ④When the OCI=4 and P/O=11 684, it means that the breeding system was facultative, outcrossing, and requiring pollinators based on the results of the bagging experiment.There was parthenogenesis. ⑤The characteristics of saponin pollination were wind pollination and insect vector pollination, and pollinators were initially identified as Apis mellifera ligustica. All these results provides a theoretical and technical foundation for the new germplasm of G. sinensis.

Separation and Characterization of Triterpenoid Saponins in Gleditsia sinensis by Comprehensive Two-Dimensional Liquid Chromatography Coupled with Mass Spectrometry.

A comprehensive two-dimensional liquid chromatography coupled with mass spectrometry method was established to separate and characterize triterpenoid saponins in Gleditsia sinensis, the Chinese herbal medicine Zhu-Ya-Zao used for the treatment of apoplexy. The saponins were separated on an Agilent Zorbax Eclipse plus C18 column (2.1 mm × 150 mm, 1.8 µm) for the first dimension, and a Poroshell 120 phenyl-hexyl column (3.0 mm × 50 mm, 2.7 µm) for the second dimension. Methanol and acetonitrile were used as the organic mobile phase for 1D and 2D, respectively. The theoretical peak capacity was 640, and the orthogonality was 57 %. Particularly, saponins with different numbers of monoterpene groups could be well separated on the second dimension. The structures were characterized by electrospray ionization tandem mass spectrometry in both positive and negative ion modes. In the MS/MS spectra, the relative abundances for [B α + Na]+ and [Y 0α + Na]+ were closely correlated with the number of sugar residues of the α-chain and ß-chain, respectively, which facilitated the differentiation of isomers. Finally, a total of 72 saponins with molecular weights greater than 1500 Da were characterized. Among them, 49 compounds, including 2 acetylated saponins, were detected from G. sinensis for the first time.

De novo assembly and characterization of Gleditsia sinensis transcriptome and subsequent gene identification and SSR mining.

Gleditsia sinensis is a Chinese native deciduous tree with a high economic and medicinal value. However, there is limited knowledge on the molecular processes responsible for the medical properties of this species owing to lack of bioinformatic resources such as available whole-genome sequences. In the present study, RNA sequencing data were used to analyze the transcriptome of G. sinensis, and a series of bioinformatic tools was used to explore the main genes involved in important molecular processes. A total of 75.57 million paired-end reads, with a length of 101 bp, were acquired from G. sinensis. Using the assembly tool Trinity, 233,751 transcripts were discovered. Among these, 85,795 were identified as unique transcripts and 59,326 unique transcripts were found to contain coding regions. Gene ontology analysis identified 27,637 unique transcripts that were clustered into 56 functional groups. Genes involved in flavonoid and terpenoid backbone biosynthesis and those encoding transcription factors were further analyzed. Sequence analysis revealed four putative G. sinensis chalcone isomerase genes (GsCHI) encoding the enzymes for flavonoid biosynthesis. GsCHI1 was found to be phylogenetically related to the chalcone isomerase of the family Leguminosae, and its transcript levels in different tissues were higher than those of GsCHI2, GsCHI3, and GsCHI4. Furthermore, 15,014 simple sequence repeat (SSR) markers were discovered in the transcript library, and 5170 primers were generated for the SSR loci. The genetic and genomic information presented in this study will be helpful for future studies on gene discovery and molecular processes in G. sinensis.

Gleditsia sinensis Thorn Attenuates the Collagen-Based Migration of PC3 Prostate Cancer Cells through the Suppression of α2ß1 Integrin Expression.

Gleditsia sinensis thorns (GST) have been used as a traditional medicine for carbuncles and skin diseases. The purpose of this study was to decide whether non-toxicological levels of water extract of GST (WEGST) are effective in inhibiting the progress of prostate cancer formation and to identify the target molecule involved in the WEGST-mediated inhibitory process of prostate cancer cell migration and in vivo tumor formation. Through the Boyden chamber migration assay, we found that non-toxic levels of WEGST could not attenuate the PC3 migration to the bottom area coated with serum but significantly inhibited PC3 cell migration to the collagen-coated bottom area. We also found that non-toxic levels of WEGST significantly attenuated collagen against adhesion. Interestingly, ectopic administration of WEGST could not affect the expression of α2ß1 integrin, which is known as a receptor of collagen. However, when the PC3 cells adhered to a collagen-coated plate, the expression of α2 integrin but not that of ß1 integrin was significantly inhibited by the administration of non-toxic levels of WEGST, leading to the inhibition of focal adhesion kinase (FAK) phosphorylation. Furthermore, oral administration of WEGST (25 mg/kg/day) significantly inhibited the size of a PC3 cell-xenografted tumor. Taken together, these results suggest a novel molecular mechanism for WEGST to inhibit prostate cancer progression at particular stages, such as collagen-mediated adhesion and migration, and it might provide further development for the therapeutic use of WEGST in the treatment of prostate cancer progression.

Hypoglycemic activity of Gleditsia caspica extract and its saponin-containing fraction in streptozotocin-induced diabetic rats.

The fruits of Gleditsia species (Fabaceae) have been known in traditional medicine as a saponin-rich herbal medicine. The present study aimed to investigate the effects of the total methanolic extract of Gleditsia caspica (MEGC) and its saponin-containing fractions (SFGC) on hyperglycemia in streptozotocin (STZ)-induced diabetic rats. A single intraperitoneal injection of STZ (55 mg/kg body weight) was used to induce hyperglycemia in male albino rats. MEGC (15, 30 and 60 mg/kg, p.o.) and SFGC (15, 30 and 60 mg/kg, p.o.) were administered to the diabetic rats daily for 14 days. The anti-diabetic drug gliclazide (10 mg/kg, p.o.) was used as a positive control. Blood samples were collected from overnight fasted rats for the evaluation of the antihyperglycemic, antihyperlipidemic and antioxidant activities. The levels of glucose, triglycerides (TG), cholesterol (TC) and malondialdehyde (MDA) were increased significantly, whereas the levels of α-amylase, insulin and reduced glutathione (GSH) were decreased in the experimental diabetic rats. Pancreas and liver of the diabetic rats exhibited significant changes in the histopathology, morphology and DNA content. Administration of MEGC or SFGC led to a decrease in the levels of glucose, TG, TC and MDA. In addition, the levels of α-amylase, insulin and GSH were increased in MEGC and SFGC treated diabetic rats. Also, the histopathological and morphological changes, as well the changes in DNA were significantly reversed by the extracts. Thus, MEGC and SFGC exhibited potent hypoglycemic and hypolipidemic activities in STZ- induced diabetic rats.

Characterization of Fractional Polysaccharides from Gleditsia sinensis and Gleditsia microphylla Gums.

The seeds of Gleditsia sinensis and Gleditsia microphylla, widespread in China, are an important source of galactomannans. G. sinensis gum (GSG) and G. microphylla gum (GMG) were purified and precipitated using different concentrations of ethanol and isopropanol. The GSG and GMG, precipitated in different stages, presented different characteristics, including polymer recovery, mannose/galactose ratio, chemical composition, molecular weight, and morphological appearance. The galactomannan recovery of GSG and GMG in 33.3% ethanol was 81.7% and 82.5%, respectively, while that in 28.8% isopropanol was 81.3% and 82.9%, respectively. To achieve similar precipitation efficiency, the amount of isopropanol should be lower than that of ethanol because of the lower dielectric constant of isopropanol (20 vs. 25 for ethanol). The precipitation behavior of galactomannans in polar organic solvents was dependent on the molecular structures and properties of the solvent. A higher mannose/galactose ratio and a higher molecular weight was obtained in a lower concentration of alcohols.

Analgesic activity of Gleditsia triacanthos methanolic fruit extract and its saponin-containing fraction.

CONTEXT: Gleditsia triacanthos L. (Leguminosae) pods are used in folk medicine for pain relief as anodyne and narcotic. OBJECTIVE: The objective of this study is to evaluate analgesic activity of Gleditsia triacanthos methanolic fruit extract (MEGT) and its saponin-containing fraction (SFGT). MATERIALS AND METHODS: Peripheral analgesic activity was assessed using the acetic acid-induced writhing model in mice at doses of 140, 280, and 560 mg/kg and formalin test in rats at 100, 200, and 400 mg/kg doses. Central analgesic activity was evaluated using the hotplate method in rats (100, 200, and 400 mg/kg). RESULTS: In the writhing test, six mice groups treated with MEGT and SFGT found ED50 values 268.2 and 161.2 mg/kg, respectively, displayed a significant decrease in writhing count compared with the group treated with standard drug indomethacin (14 mg/kg). SFGT (280 and 560 mg/kg) showed 64.94 and 70.78% protection, respectively, which are more than double % protection caused by indomethacin (31.82%). In the formalin test, MEGT and SFGT (ED50 values 287.6 and 283.4 mg/kg for phase I as well as 295.1 and 290.4 mg/kg for phase II, respectively) at 400 mg/kg showed significant % inhibition in both phase I (18.86 and 52.57%) and phase II (39.36 and 44.29%) with reference to 10 mg/kg indomethacin (56.0 and 32.29%). MEGT and SFGT caused significant delay in responses in hotplate model (ED50 values 155.4 and 200.6 mg/kg, respectively) compared with that of 10 mg/kg indomethacin at 30, 60, and 120 min. DISCUSSION AND CONCLUSION: Central and peripheral analgesic activities induced by Gleditsia triacanthos fruits might account for its uses in folk medicine.

In Vivo Anti-tumor Effects of the Ethanol Extract of Gleditsia sinensis Thorns and Its Active Constituent, Cytochalasin H.

Angiogenesis is the process of new vessel formation from pre-existing blood vasculature and is critical for continuous tumor growth. We previously reported that an ethanolic extract of Gleditsia sinensis thorns (EEGS) and its active constituent, cytochalasin H, have anti-angiogenic activity in vitro and in vivo via suppression of endothelial cell functions. In the present study, EEGS and cytochalasin H were observed to efficiently inhibit tumor growth in an in ovo xenograft model without significant toxicity. We repeatedly observed the anti-tumor and anti-metastatic effects of EEGS in representative animal models. These results suggest that EEGS and its active constituent, cytochalasin H, are potential candidates for the development of anti-angiogenic cancer drugs.