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As a medicinal herbal plant, Entada phaseoloides has high levels of secondary metabolites, particularly triterpenoid saponins, which are important resources for scientific research and medical applications. However, the lack of a reference genome for this genus has limited research on its evolution and utilization of its medicinal potential. In this study, we report a chromosome-scale genome assembly for E. phaseoloides using Illumina, Nanopore long reads and high-throughput chromosome conformation capture technology. The assembled reference genome is 456.18 Mb (scaffold N50 = 30.9 Mb; contig N50 = 6.34 Mb) with 95.71% of the sequences anchored onto 14 pseudochromosomes. E. phaseoloides was estimated to have diverged from the Leguminosae lineage at ~72.0 million years ago. With the integration of transcriptomic and metabolomic data, gene expression patterns and metabolite profiling of E. phaseoloides were determined in different tissues. The pattern of gene expression and metabolic profile of the kernel were distinct from those of other tissues. Furthermore, the evolution of certain gene families involved in the biosynthesis of triterpenoid saponins and terpenes was analysed and offers new insights into the formation of these two metabolites. Four CYP genes, one UGT gene and related transcription factors were identified as candidate genes contributing to regulation of triterpenoid saponin biosynthesis. As the first high-quality assembled reference genome in the genus Entada, it will not only provide new information for the evolutionary study of this genus and conservation biology of E. phaseoloides but also lay a foundation for the formation and utilization of secondary metabolites in medicinal plants.

The conventional method for effective or toxic chemical substance identification of multicomponent herbal medicine is based on single component separation, which is time-consuming, labor intensive, inefficient, and neglects the interaction and integrity among the components; therefore, it is necessary to find an alternative routine to evaluate the components more efficiently and scientifically. In this study, sodium aescinate injection (SAI), obtained from different manufacturers and prepared as “components knockout” samples, was chosen as the case study. The chemical fingerprints of SAI were obtained by high-performance liquid chromatography to provide the chemical information. The effectiveness and irritation of each sample were evaluated using anti-inflammatory and irritation tests, and then “Gray correlation” analysis (GCA) was applied to rank the effectiveness and irritability of each component to provide a preliminary judgment for product optimization. The prediction model of the proportions of the expected components was constructed using the artificial neural network. The results of the GCA showed that the irritation sorting of each SAI component was in the order of B > A > G > J > I > H > D > F > E > C and the effectiveness sorting of SAI components was in the order of D > C > B > A > F > E > H > I > G > J; the predictive proportion of SAI was optimized by the BP neural network as A: B: C: D: E: F = 0.7526: 0.5005: 5.4565: 1.4149: 0.8113: 1.0642. This study provided a scientific, accurate, reliable, and efficient approach for the proportion optimization of multicomponent drugs, which has a good prospect of popularization and application in product upgrading and development of herbal medicine.

Six new serratene triterpenoids (1-6), together with nine known triterpenoid compounds were isolated from the extract of club moss Lycopodium japonicum. The structures of isolated compounds were established by spectroscopic methods. The cytotoxic activities of all compounds were evaluated against three human cancer cell lines in vitro by MTT assay. Compounds 2, 6-8 and 11 exhibited moderate activities against all three cell lines with IC50 values of 2.28-11.81 μg/mL.

Glycoborinine (GB), a natural carbazole alkaloid isolated from Glycosmis pentaphylla, has been shown to be a potential molecule against cancer cells. In this study, the cell-signaling pathway of its anti-tumor activity was investigated. MTT assay result showed that GB inhibited HepG2 cell proliferation in a dose- and time-dependent manner and 50% inhibiting concentration (IC50) of GB-induced cell death was 39.7 μM for a period of 48 h. GB-induced HepG2 apoptosis was confirmed by Hochest 33258 staining and PI staining. The level of reactive oxygen species (ROS) was measured with H2DCF-DA staining and the change of mitochondrial membrane potential (△Ψ(m)) was analyzed with tetrechloro-tetraethylbenzimidazolcarbocyanine iodide (JC-1) probe. Results showed that GB at 12.5, 25, and 50 μM promoted ROS production. GB induced HepG2 apoptosis through a mitochondrial apoptotic pathway, which was demonstrated by GB-induced increase in the ratio of Bax/Bcl-2, cytochrome C release, the ratio of cleaved caspase-3/procaspase-3, and the ratio of cleaved poly ADP-ribose polymerase (cleaved PARP)/poly ADP-ribose polymerase (PARP). To summarize, this study demonstrated that GB could induce HepG2 apoptosis through the mitochondrial-dependent pathway, which might provide a promising approach to cure liver cancer with GB.

The leaves of Elaeagnus pungens were extracted with 70% ethanol and successively purified by column chromatography. Seven constituents were obtained and characterized, all of which belong to the class of flavonol glycosides. Their structures were elucidated on the basis of spectroscopic methods including 1D/2D NMR and MS analysis techniques. The seven flavonol glycosides were determined as kaempferol 3-O-β-d-glucopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 6)-[α-l-rhamnopyranosyl(1 → 2)]-β-d-galactopyranoside (1), isorhamnetin 3-O-β-d-glucopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 6)-β-d-galactopyranoside (2), together with five known compounds, respectively. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliu-m bromide assay in vitro showed that the isolated flavonol glycosides showed no proliferation activity in the asthma airway smooth muscle cells, comparing with solvent as the control group.

Two new phenylethanoid glycosides, himalosides A (1) and B (2), were isolated from the whole plant of Boschniakia himalaica, together with four known compounds. Their structures were elucidated on the basis of spectral and chemical evidences. Himaloside A (1) and (+)-lariciresinol 4′-O-β- d -glucopyranoside (4) exhibited antibacterial activity with the MIC values at 50 and 25 μg/mL against Staphylococcus aureus and S. epidermidis, respectively.

Three new C21 steroidal glycosides with a cinnamoyl group at C(12) and a 2-methylbutanoyl group at C(20), and a straight sugar chain at C(3), namely cyanoauriculosides C–E (1–3, resp.), together with three known steroidal derivatives, were isolated from the roots of Cynanchum auriculatum (Asclepiadaceae). Their structures were determined by spectroscopic analyses and chemical methods. The known constituents were identified as wilfoside K1N (4), cynanauriculoside II (5), and auriculoside IV (6).

Three new macrocyclic β-dihydroagarofuran-type sesquiterpene pyridine alkaloids, fortuneines A (1), B (2), and C (3), together with the four known alkaloids wilfornine E (4), aquifoliunine E-I (5), euoverrine B (6), and euojaponine I (7), were isolated from the aerial parts of Euonymus fortunei. Their structures were elucidated by spectroscopic methods, including HR-ESI-MS, 1H- and 13C-NMR, DEPT, 1H,1H-COSY, HSQC, HMBC, and ROESY. This is the first isolation of the above sesquiterpene pyridine alkaloids from this plant, except for compound 6.

Two new steroidal glycosides named cyanoauriculoside A (1) and B (2), have been isolated from the roots of Cynanchum auriculatum Royle ex Wight. The structure of cyanoauriculoside A (1) was determined to be 20-O-acetyl-penupogenin-3-O-β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl-(1→4)-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-diginopy-ranosyl-(1→4)-β-D-cymaropyranoside. Cyanoauriculoside B (2) was characterized as 20-O-acetylpenupogenin-3-O-β-D-glucopyranosyl-(1→4)-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-diginopyranosyl-(1→4)-β-D-cymaropyranoside. The structures of these compounds were elucidated based on the chemical and spectroscopic evidence.

The CO I gene sequences of Qianghuoyu, Pachytriton labiatus and Gehyra mutilata were achieved by PCR amplification and bi-directional sequencing. Furthermore, a pair of specific primers SJYW1 and SJYW2 in the non-conservative district were designed through sequence alignment. The PCR reaction condition was established by changing the annealing temperature and cycle numbers. The results showed that 350 bp DNA fragment was amplified from Qianghuoyu in PCR with annealed temperature at 54 °C and the cycle number was 25 cycles, whereas not any DNA fragment was amplified from P. labiatus and G. mutilata under the same reaction condition. This method is well-performed in the identification of Qianghuoyu for its excellent specificity and repeatability.

Two novel xanthones, 1,6-dihydroxy-4,5-dimethoxyxanthone ( 1 ) and 1,5,6-trihydroxy-7,8-di(3-methyl-2-butenyl)-6′,6′-dimethylpyrano(2′,3′:3,4)xanthone ( 2 ) were isolated from the bark of Garcinia xanthochymus by normal phase and reverse phase silica gel column chromatography. Their structures were elucidated by spectroscopic methods, especially 2D-NMR techniques.

In the title compound, C15H22O2, the C=C bond lengths in the cyclohexene ring and the allylic acid group [1.325 (3) and 1.324 (3) Å] are almost equal and are shorter than those observed in ethylene. The C=O and C—O bond lengths in the allylic acid group [1.201 (3) and 1.321 (3) Å] are almost equal to those of formic acid. There is an intramolecular hydrogen bond between the allylic acid H atom and the hydroxy O atom [C...O = 2.673 (2) Å and C—H...O = 101°].

OBJECTIVE: To study water-soluble chemical constituents from the leaves of Elaeagnus pungens. METHOD: Chemical constituents of E. pungens leaves were separated by a combination of macroporous resin column chromatography, reverse phase silica gel column chromatography, Sephadex LH-20 column chromatography and semi-preparative HPLC. Their structures were identified on the basis of physicochemical properties using the spectral method. RESULT: The two compounds were separated from E. pungens leaves and identified as kaempferol 3-O-P-D-glucopyranosyl- (1-->3)-alpha-L-rhamn-opyranosyl-(1-->6) -/3-D-galactopyranoside (1), kaempferol 3-O-P-D-glucopyranosyl-(1-->3)-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranoside-7-O-beta-D-glucopyranoside (2). CONCLUSION: Compound 2 separated from E. pungens leaves was a new compound.

To study identification methods of different extracts of Dai Medicine "Pokou" (the rhizome of Homalomena gigantea Engl. ) and its processed product made by immersing it in water, and provide reference for identification of the drug in further researches and applications.FTIR technique was used for identifying the features of different extracted parts of this crude drug and its processed product.Compared with the crude drug, the petroleum ether-extracted parts of processed product turned out to have no obvious distinction in the FTIR. There was a large difference in the ethyl acetate-extracted parts, and the n-butanol-extracted parts also had certain discrepancy. A preliminary analysis was made on the chemical fundamentals which caused the changes in the FTIR before and after the drug's processing.The results provide an infrared spectral identification method for the drug and its applications.

To study the acute toxicity of the crude and processed products of Entada phaseoloides and their effects on gastrointestinal movement in mice.Using the method of intragastric administration, to observe the acute toxicity of the crude and processed products of Entada phaseoloides in mice and determine their LD50. With the methods of charcoal propulsion of small intestine and methyl orange colorimetry of gastric emptying, to study the impact of the crude and processed products of Entada on gastrointestinal movement in mice.The oral LD50 of crude Entada phaseoloides, No. 1 and No. 2 processed products of Entada phaseoloides in mice were 27.17, 35.13, 42.18 g/kg body weight. Crude and processed products of Entada phaseoloides can significantly promote the enteric propulsion of normal mice, and can significantly counteract the depressing status induced by atropine, but have no influence on the overactive status induced by neostigmine. The high, middle and low-dose of groups showed significant inhibition of the gastric emptying in normal mice.Processed Entada phaseoloides showed effects on the enteric propulsion of normal and depressing mice, can restrain the gastric emptying under normal mice, but its safety is better than crude Entada phaseoloides.

To study the chemical constituents from the root of Berchemia lineata (L.) DC., nine compounds were isolated from the EtOAc extract by using silica gel, RP-C18 silica gel column chromatography and preparative HPLC. Based on the spectroscopic analysis, their structures were identified as 5-hydroxy-7-(2'-hydroxypropyl)-2-methyl-chromone (1), (-)-(1'R, 2'S)-erythro-5-hydroxy-7-(1', 2'-dihydroxypropyl)-2-methyl-chromone (2), naringenin (3), eriodictyol (4), (+)-aromadendrin (5), (+)-taxifolin (6), (+)-catechin (7), (+)-epigallocatechin (8) and quercetin (9). Among them, compound 2 is a new chromone derivative. Compound 1 is a known chromone derivative and isolated from this genus for the first time. Compounds 3-9 are known flavonoids and isolated from this plant for the first time.

To research identification methods of the Dai Medicine "Pokou" (the rhizome of Homalomena gigantea) and its processing product, and provide basis for identification of the drug in further research and application.Macroscopic, microscopic observation and TLC and FTIR techniques were used to authenticate this raw medicine and its processing product.There were certain differences in the macroscopic features. The TLC result and infrared spectra of the samples had also obvious differences. The methods for identification of this raw medicine and its processing product were established, The detailed tissue and powder of this medicine were drawn.The results provided the basis for identification of the medicine and establishment of its quality standard.

OBJECTIVE A novel method was established and the results were compared for HPLC fingerprint determination of crude and processed products of Entada phaseoloides. METHOD HPLC-UV was proposed for studying the fingerprints of fresh E. phaseoloides and their processed products, respectively. HPLC-ESI-MS was introduced to analyze the common peaks in each batch of crude E. phaseoloides. RESULT Sixteen characteristic peaks were found in crude E. phaseoloides samples and twenty-one common peaks existed in processed E. phaseoloides samples. Nine characteristic peaks of which were identified by comparison of the retention time and their molecular weights of chemical standards, most of which were identificated belong to triterpenoid saponins and glucosides. CONCLUSION After processed, the chemical composition of the extraction with solution of 60% methanol from crude E. phaseoloides are less or more than that from processed E. phaseoloides, and the changes of the main peaks of fingerprint chromatographic suggest that HPLC can be used to reflect the difference of chemical composition of E. phaseoloides and their processed products. It would be an efficient way for qualitative control of E. phaseoloides.

Three new steroidal glycosides, cyanoauriculosides F, G and H (1-3), were isolated from the roots of Cynanchum auriculatum (Asclepiadaceae) along with two known steroidal derivatives. On the basis of spectroscopic analysis and chemical methods, their structures were identified as 20-O-acetyl-8,14-seco-penupogenin-8-one 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-diginopyranosyl-(1→4)-β-D-cymaropyranoside (1), 2′,3′-Z-gagaminine 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaro-pyranosyl-(1→4)-α-L-diginopyranosyl-(1→4)-β-D-cymaropyranoside (2), 17-O-acetyl-kidjoranin 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-cymaro-pyranosyl-(1→4)-β-D-digitoxopyranosyl-(1→4)-β-D-digitoxopyranoside (3), gagaminine 3-O-α-L-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-α-L-digino-pyranosyl-(1→4)-β-D-cymaropyranoside (4) and wilfoside D1N (5).

To study the chemical constituents of the Entada phaseoloides (L.) Merr., seeds of Entada phaseoloides were extracted with 70% ethanol at room temperature. Isolation and purification were performed by silica gel, reversed-phase silica gel column chromatography and semi-preparative HPLC. Structures of the pure compounds were established on the basis of spectral analysis. Four sulfur-containing amide compounds were isolated from the n-BuOH-soluble fraction and identified as entadamide A-beta-D-glucopyranosyl-(1--3)-beta-D-glucopyranoside (1), entadamide A (2), entadamide A-beta-D-glucopyranoside (3) and clinacoside C (4). Compound 1 is a new compound. Compound 4 is isolated from the genus Entada for the first time.

To investigate the chemical constituents of the aerial parts of Plumbago zeylanica Linn.The constituents of the EtOAc-soluble portion in the 95% ethanol extract were isolated and purified by means of chromatography. Compounds were identified by their physical characteristics and spectral features.Nine compounds were isolated as plumbagin (I), isoshinanolone (II), plumbagic acid (III), beta-sitosterol (IV), 4-hydroxybenzaldehyde (V), trans-cinnamic acid (VI), vanillic acid (VII), 2, 5-dimethyl-7-hydroxychromone (VIII), indole-3-carboxaldehyde (IX).Compounds V, VII, VIII and IX were isolated for the first time from Plumbago Linn.

To study the chemical constituents of aerial parts of Laggera pterodonta (DC.) Benth., the air-dried aerial parts of this plant were powered and extracted with boiling water and purified by silica gel column chromatography and recrystallized. Eleven compounds were obtained from L. pterodonta. They were identified as to be 6-O-beta-D-glucopyranosyl-carvotanacetone (1), pterodontic acid (2), 1beta-hydroxy pterondontic acid (3), pterodontoside A (4), pterodondiol (5), pterodontriol B (6), 5-hydroxy-3,4', 6,7-tetramethoxyflavone (7), artemitin (8), chrysosplenetin B (9), quercetin (10) and beta-sitosterol (11). Compound 1 is a new monoterpene glucoside. Compounds 10 and 11 were isolated from this plant for the first time. Compounds 2 and 5 showed moderate activity against bacteria including Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Mycobacteium phlei and Bacillus circulans by paper disc diffusion method, while they both displayed no activity against Escherichia coli.

To investigate the antihepatocellular carcinoma effects of Chi-Shen extract (CSE) from the water-soluble compounds of Salvia miltiorrhiza and Paeoniae radix. The effect of CSE on the growth of HepG2 cells (hepatocellular carcinoma cell line) was studied by 3-(4,5)-2,5-diphenyltetrazolium bromide assay. Apoptosis were detected through acridine orange (AO) and ethylene dibromide (EB) staining and DNA fragmentation assay. The effect of CSE on the cell cycle of HepG2 cells was studied by the propidium iodide staining method. The activation of caspases-3, -8 and -9 was examined by immunoassay kits. The transcription of the Bcl-2 family and p53 was detected by RT-PCR. Our data revealed that CSE strongly induced HepG2 cell death in a dose- and time-dependent manner. CSE-induced cell death was considered to be apoptotic by observing the typical apoptotic morphological change by AO/EB staining and DNA fragmentation assay. The induction of HepG2 cell death was caused by an induction of apoptosis for the sub-G1 proportion increase, the downregulation of Bcl-2, the upregulation of Bax and p53, and the activation of the caspases-3 and -9 pathways. These results clearly demonstrated that CSE was able to inhibit the proliferation of HepG2 cells and cause apoptosis. Moreover, the anticancer effects of CSE were related to the Bcl-2 family pathway and the activation of caspases-3 and -9 in HepG2 cells.