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Objective To establish multi-class bioactive constituents’ determination of ten Anoectochilus, four Goodyera and one Ludisia species, and provide reference for the improvement of their quality control. Methods HPLC-ELSD and phenol–sulphuric acid methods were used for the quantitative determination of lactone glycosides (kinsenoside and its diastereoisomer, goodyeroside A) and polysaccharides, respectively, while an efficient iHPLC–MS/MS method was established for rapid determination of other minor constituents in ten Anoectochilus species and five related species. Results The contents of kinsenoside, goodyeroside A, polysaccharides and flavonoids varied notably almost in all tested samples, including both wild plants and tissue cultures. In particular, kinsenoside was the major lactone glycoside in A. roxburghii, A. formosanus, A. xingrenensis, A. nandanensis, A. brevilabris and A. burmannicus, whereas goodyeroside A was the predominant constituent in A. lylei, A. longilobus, A. elatus, A. zhejiangensis, G. schlechtendaliana, G. biflora, G. yangmeishanensi, G. repens and Ludisia discolor. Conclusion Our present study suggested that A. lylei, A. longilobus, A. elatus, A. zhejiangensis, Ludisia discolor and Goodyera species cannot be used as alternatives for A. roxburghii, and goodyeroside A may be reasonably used as a diagnostic marker for distinguishing A. roxburghii from A. lylei, A. longilobus, A. elatus and A. zhejiangensis, Goodyera and Ludisia species. The established method thus could be potentially used for the quality evaluation and control of Anoectochilus and some related species.

We recently proposed, and successfully applied, a novel and efficient technique-ultrasonic-circulating extraction (UCE) integrating superfine pulverization-to extract and prepare antioxidant crude polysaccharides other natural active substances from Ganoderma lucidum. The aim of this study was to evaluate the antioxidant and hepatoprotective activities and active ingredients in the powder from UCE (UCEP) through comparison with powder from hot water extraction (HWEP). The DPPH radical, ABTS radical, superoxide anion, total antioxidant activity, and ferric-reducing antioxidant power assay results showed that the UCEP exhibited stronger (P < 0.01) in vitro antioxidant activity than the HWEP. The hepatoprotective activity of the extracts was evaluated against CCl4-induced oxidative damage in the liver. Measurements of reduced glutathione, superoxide dismutase, and malondialdehyde in rat liver; measurements of alanine transaminase, aspartate transaminase, and lactate dehydrogenase in rat blood; and Western blotting for antioxidant proteins of transforming growth factor-β1, heme-oxygenase 1, and glutathione per-oxidase showed that the UCEP had antioxidant activity in vivo either similar to or slightly stronger than (P < 0.1) the HWEP. Further analysis of the active ingredients revealed that the UCEP and HWEP have similar mean yield and total triterpenoid content, but the former has significantly higher (P < 0.05) mean yield and total polysaccharide content than the latter. Our results suggest that the UCEP displays stronger antioxidant activities because of the larger amount of total polysaccharides; the UCEP may be able to be used as an antioxidant and liver protectant.

Ethnopharmacological relevance The roots of Actinidia eriantha Benth (AER) are commonly used traditional folk medicine for the treatment of gastric carcinoma, nasopharyngeal carcinoma, and breast carcinoma. Besides, the anti-proliferative and immunomodulatory effects of AER polysaccharides on tumor-bearing mice have been reported previously. Aim of the study This work was carried out to investigate the anticancer and anti-angiogenic activities of AER. Materials and methods The growth inhibitory effects of ethanol extracts from the leaves (EEL), stems (EES) and roots (EER) of A. eriantha on human gastric carcinoma SGC7901 cells, human nasopharyngeal carcinoma CNE2 cells, human breast carcinoma MCF7 cells and human umbilical vein endothelial cells (HUVECs) were evaluated by MTT assay. The ethyl acetate fraction from EER (EA-EER) was further investigated for the anticancer activity against SGC7901 cells and the anti-angiogenic activity in HUVECs in vitro . The apoptosis in SGC7901 cells and HUVECs was confirmed by DAPI nuclear staining and flow cytometry analysis, the effect on cellular DNA fragmentation was detected in SGC7901 cells. And the cell cycle-arresting activity in HUVECs was determined by flow cytometry. Moreover, the inhibitory effect of EA-EER on cell migration in HUVECs was observed by both wound-healing and Transwell migration assays. RT-PCR and Western-blotting were performed to determine the mRNA and protein expression levels, respectively, including Bax, Bcl-2 and caspase-3 in SGC7901 cells, as well as VEGF-A and VEGFR-2 in HUVECs. Furthermore, the in vivo anti-angiogenic activity of EA-EER was evaluated by using chick embryo chorioallantoic membrane (CAM) assay. Ultimately, the chemical components in EA-EER were isolated and purified by repeated column chromatography followed by structure characterization using 1 H and 13 C nuclear magnetic resonance and mass spectroscopy. Results Compared with EEL and EES, EER displayed the strongest growth inhibitory effect on SGC7901 cells, CNE2 cells and HUVECs. Among the EER fractions, EA-EER exhibited the most potent growth inhibitory activity against SGC7901 cells, CNE2 cells and HUVECs. Moreover, EA-EER induced obvious apoptosis in SGC7901 and HUVECs, and significantly inhibited the proliferation of HUVECs via blockade of cell cycle G1 to S progression. Furthermore, EA-EER suppressed the expression of Bcl-2 and improved the expression Bax and caspase-3 in SGC7901 cells. EA-EER not only inhibited migration of HUVECs, but also down-regulated the expression of VEGF-A and VEGFR-2 in HUVECs. In vivo, EA-EER exposure reduced the formation of blood vessels in chick embryos. A bio-guided isolation of EA-EER led to the isolation of three compounds for the first time, namely (6R, 7E, 9S)-6, 9-hydroxy-megastigman-4, 7-dien-3-one-9-O-β- D -glucopyranoside, Oleanolic acid-23-O-β- D -glucopyranoside, 3β, 23, 24-trihydroxyl-12-oleanen-28-oic acid. Conclusion The present research demonstrated that the significant anticancer and anti-angiogenic effects of AER, providing the supportive evidence for its traditional use in the treatment for cancer. It was suggested that AER could be use as a potential source of cancer therapeutic drug.

The high-performance liquid chromatography fingerprint method is a simple and reliable technique to evaluate the quality of leaves of Magnolia officinalis Rehd.et Wils. var. biloba Rehd.et Wils. We used the following bioactive phenolic constituents as reference compounds: rutin, afzelin, hyperoside, isoquercitrin, quercetin-3-O-α-l-rhamnoside, honokiol and magnolol. The conditions of an Agilent 1200 HPLC were: YMC-Pack-ODS-AQ column (250 × 4.6 mm id S-5 μm, 12 nm), mobile phase acetonitrile and 0.2% phosphoric acid in a gradient elute mode, flow rate 1.0 mL/min, detection wavelength 280 nm and column temperature 30°C. The analytical method was validated in terms of linearity, stability, repeatability, precision and recovery tests. While performing fingerprint analysis, we identified 11 peaks as characteristic peaks and assessed the similarities of 17 samples collected from different geological regions of China. The peak areas were used to evaluate the variation in the chemical composition of the tested samples. For this purpose, we performed hierarchical cluster analysis of the peak areas. Our results indicate that simultaneous determination of multiple ingredients could be done through chromatographic fingerprint analysis. Therefore, this high-performance liquid chromatography fingerprint method was readily utilized to evaluate the quality of leaves of M. officinalis var.biloba, which are used in several traditional herbal preparations.

Background Iridoid glycosides (IGs), including monotropein (MON) and deacetyl asperulosidic acid (DA) as the main ingredients, are the major chemical components in Morinda officinalis How. (MO) root, possessing various pharmacological properties including anti-osteoporosis, anti-inflammation and anti-rheumatism activities.The aim of the present study was to further elucidate the pharmacological actions of MO by investigating the pharmacokinetics and tissue distribution of IGs in MO. Methods An ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS) method was developed and validated for simultaneous determination of MON and DA levels in plasma and various tissues of Wistar rats. MON, DA and acetaminophen (ACE) as the internal standard (IS) were extracted from rat plasma and tissue samples by direct deproteinization with methanol. The rats were administered orally at 1650 mg/kg MO and 25, 50 and 100 mg/kg MO iridoid glycosides (MOIGs) or intravenously at MOIG 25 mg/kg for pharmacokinetic study of MON and DA. In addition, 100 mg/kg MOIG was administered orally for tissue distribution study of MON and DA. Non-compartmental pharmacokinetic profiles were constructed. Tissue distributions were calculated according to the validated methods. Results Significant differences in the pharmacokinetic parameters were observed in male and female rats. The AUC0-t, Cmax and bioavailability of MON and DA in female rats were higher than those in male rats. MON and DA mainly distributed in the intestine and stomach after oral administration, and noteworthily high concentrations of MON and DA were detected in the rat hypothalamus. Conclusion The results of the present study may shed new lights on the biological behavior of MOIGs in vivo, help explain their pharmacological actions, and provide experimental clues for rational clinical use of these IGs extracted from the MO root. Electronic supplementary material The online version of this article (10.1186/s12906-018-2351-1) contains supplementary material, which is available to authorized users.

Chemical investigation on the solid rice culture of Trichoderma atroviride S361, an endophyte isolated from Cephalotaxus fortunei, has afforded a pair of novel N-furanone amide enantiomers, (-)-trichodermadione A (1a) and (+)-trichodermadione A (1b), and a new cyclohexenone sesquiterpenoid, trichodermadione B (2), together with six known secondary metabolites. Chiral separation of compound 1 was successfully performed on a Lux Cellulose-2 column. Their structures were elucidated by detailed spectroscopic analyses on the basis of NMR, HRMS, and ECD data, and the absolute configurations of the new compounds were determined by computational analyses of their electronic circular dichroism (ECD) spectra and Snatzke's method. Compounds 1a, 1b and 2 were also evaluated for their cytotoxicity against DU145 and PC3 cell lines, as well as inhibitory effects against the production of NO in LPS-stimulated RAW264.7 cells.

Objective To evaluate the effects of Jidesheng anti-venom used externally for skin and soft-tissue necrosis from Chinese cobra bite. Methods A retrospective review was performed according to the clinical data recorded from January 2002 to December 2012. A total of 126 patients (116 females and 10 males) with skin and soft-tissue necrosis due to Chinese cobra bite were divided into two groups. The control group was treated externally with 40% glyceride magnesium sulfate ( n =52), and the treatment group was given Jidesheng anti-venom externally ( n =74). The data collected included maximum local necrotic area of skin and soft tissues, de-tumescence time, healing time, and skin-grafting rate. Results There were no significant differences in gender, age, and disease condition between the control and treatment groups ( P >0.05). No statistically significant difference was found in de-tumescence time between the two groups ( P >0.05). The maximum local necrotic area of skin and soft tissues was (19.9 ± 7.3) cm 2 in the treatment group, while it was (23.3 ± 6.4) cm 2 in the control group. The healing time of the treatment group was shorter than that of the control group [(32.1 ± 3.7) vs (34.4±4.5) days)] The skin-grafting rate in the treatment group was lower than that of the control group (10.81% vs 25.00%). There were statistically significant differences in maximum local necrotic area of skin and soft tissues, healing time, and skin-grafting rate between the control and treatment groups (all P Conclusion External application of Jidesheng anti-venom may help to promote wound healing and reduce the skin-grafting rate in cases of skin and soft-tissue necrosis due to Chinese cobra bite.

A reversed-phase high performance liquid chromatography (HPLC) method was developed to evaluate the quality of Morinda officinalis through establishing chromatographic i¬ngerprint and simultaneous determination of four anthraquinone compounds, namely 2-hydroxy-3-hydroxymethyl- anthraquinone, 2-hydroxy-1-methoxy-anthraquinone, rubiadin-1-methyl ether and rubiadin. The chromatographic separation was performed on C18 column with a gradient elution program using a mixture of acetonitrile and 0.2% aqueous phosphoric acid (v/v) as mobile phase. In fingerprint analysis, 16 peaks were selected as the characteristic peaks to evaluate the similarities of samples collected from different origins in China according to the State Food and Drug Administration (SFDA) requirements. Furthermore, hierarchical cluster analysis (HCA) and principal component analysis (PCA) were also applied to evaluate the variation of chemical components among different sources of M. officinalis in China. This study indicated that the developed HPLC method, combined with chemometrics analysis, could be readily utilized to control the quality of M. officinalis and its related traditional Chinese medicinal (TCM) preparations. Key words: Morinda officinalis, fingerprint analysis, anthraquinone compounds, chemometric analysis, quality control.

Luteolin (LU) is considered as one of the most important flavonoids with many beneficial functions to human health. In our present study, an efficient, convenient and reproducible extraction method of LU extracted from Eclipta prostrata by ultrasound assisted microwave extraction (UAME) was established. The extraction process of LU using UAME was optimized according to single factor test and orthogonal design, and the optimum parameters were obtained as follows: the ratio of 80% ethanol, hydrochloric acid and plant material: 50: 0.3: 1 (v/v/w), microwave power: 40 W, extracting time: 3 × 3 min. The extraction yield of LU was 0.690 mg/g under the optimum conditions, compared with heat reflux extraction (HRE) and reflux acid extraction (RAE) in terms of extraction yie ld, time consumption and labor intensiveness, indicating that the UAME was more efficient than RE and RAE. In conclusion, UAME could markedly reduce extraction time and simplify the extraction process of LU fromE. prostrata. Key words: Ultrasound assisted microwave acid extraction, luteolin, Eclipta prostrata L.

The aim of this study was to evaluate antioxidant activities and cytotoxicity against human tumor cell lines of extracts from Bidens pilosa Linn. var. radiata Sch. Bip. (BP). Total phenolic and flavonoid contents of the extracts were determined by ultraviolet spectrophotometry. Furthermore, the antioxidant properties of different polarity fractions extracted from BP were evaluated using DPPH and ABTS radical scavenging test and FRAP assay. The ethyl acetate fraction (EE-BP) showed the highest antioxidant activity compared to other fractions. In addition, the anti-proliferative activities of the extracts on four human tumor cells, namely MCF-7, HepG2, MGC 803 and RKO, were investigated by MTT method. The EE-BP displayed the most remarkable anti-proliferative effect against the tumor cells, particularly RKO cell in dose- and time-dependent manner. The antioxidant activities and cytotoxicity correlated highly with the total phenolic and flavonoid contents, respectively. Furthermore, The active ingredient BP-6, namely 5,7,4'-trihydroxy-3,3'-dimethyl-flavonol, was isolated and purified with the purity above 99.00% and content of 0.15% in EE-BP detected by HPLC, which could significantly inhibit the proliferation of RKO cells with the IC(50) value of 6.66 μmol/l. In order to characterize the apoptotic RKO cells, flow cytometry and DNA fragmentation assay were performed. Apoptotic cell numbers increased in a dose-dependent manner after the treatment with different concentrations of EE-BP and BP-6 for 12 and 6 h, respectively. DNA ladders in apoptotic RKO cells could be easily visualized when exposed to 200 μg/ml of the EE-BP for 36 h. Taken together, our work indicated that BP had potentially therapeutic value against colorectal cancer.

The present study was undertaken to investigate the protective effects of ethanol extract from the root of Morinda Officinalis (RMO) on ovariectomy-induced bone loss. Administration of RMO extract increased trabecular bone mineral content and bone mineral density of tibia, improved the levels of phosphorus (P), calcium (Ca) and OPG, decreased the levels of DPD/Cr, TRAP, ACTH and corticosterone, but did not reverse the levels of ALP, TNF-alpha and IL-6 in serum of ovariectomized rats. These findings demonstrated that RMO extract reduced bone loss in ovariectomized rats, probably via the inhibition of bone resorption, but was not involved with bone formation. Anthraquinones and polysaccharides from Morinda officinals could be responsible for their antiosteoporotic activity, and the action mechanism of these constituents needs to be further studied. Therefore, RMO has the potential to develop a clinically useful antiosteoporotic agent.

Factors affecting the morphology of Streptomyces avermitilis and avermectin production in submerged cultivation, including nitrogen sources, inoculum level and DO (dissolved oxygen) tension in the broth were investigated in a 50-L bioreactor. It was found that a combination of soybean meal and yeast meal as nitrogen sources and 4.3% inoculum led to pellet formation, and the pellet morphology facilitated to maintain DO＞20% in the early stage of fermentation. With the aid of image analysis tools, area and density of pellets in different batches were calculated. Results show that higher dissolved oxygen tension was favorable for pellet formation and avermectin production.

Ganoderma triterpenoids (GTs) have been recognised as an important bioactive ingredient in Ganoderma Lucidum (Leyss. ex Fr.) Karst. (Polyporaceae), widely used for treating and preventing chronic hepatopathy of various etiologies.The objective of this study is to better understand the hepatoprotective effect of GTs and to enhance their use in food supplement pharmaceutical and medical industries.HepG2 cells were pretreated in the presence or absence of GTs (50, 100 and 200 μg/ml) for 4 h, then exposed with 60 μmol/L of t-BHP for an additional 4 h. The cell viability was evaluated by MTT method. ALT, AST and LDH production in culture medium and intracellular MDA, GSH and SOD levels were determined. Moreover, the total triterpenoid content and chemical constituents in GTs were detected by ultraviolet spectrophotometry and HPLC/Q-TOF-MS, respectively.GTs (50, 100 and 200 μg/ml) significantly increased the relative cell viability by 4.66, 7.78 and 13.46%, respectively, and reduced the level of ALT by 11.44%, 33.41% and 51.24%, AST by 10.05%, 15.63% and 33.64%, and LDH by 16.03%, 23.4% and 24.07% in culture medium, respectively. GTs could also remarkably decrease the level of MDA and increase the content of GSH and SOD in HepG2 cells. Furthermore, the total triterpenoid content in GTs was 438 mg GAAEs/g GTs. And 16 triterpenoids in GTs were identified or tentatively characterised.Our results showed that GTs had potent cytoprotective effect against oxidative damage induced by t-BHP in HepG2 cells, thus suggesting their potential use as liver protectant.

Ethnopharmacological relevance Leaves of Toona sinensis (A. Juss.) Roem. (TSL), a popular vegetable in China, have anti-inflammatory, antidoting, and worm-killing effects and are used in folk medicine for the treatment of enteritis, dysentery, carbuncles, boils, and especially abdominal tumors. Our aim was to investigate the in vitro antimicrobial activity against Staphylococcus aureus and anticancer property of the essential oil from TSL (TSL-EO), especially the pro-apoptotic effect in SGC-7901. Materials and methods TSL-EO obtained by hydrodistillation was analyzed by GC/MS and was tested in vitro against twenty clinically isolated strains of Staphylococcus aureus (SA 1–20), which were either methicillin-sensitive Staphylococcus aureus (MSSA) or methicillin-resistant Staphylococcus aureus (MRSA) and two standard strains viz. ATCC 25923 and ATCC 43300. The anticancer activity of TSL-EO was evaluated in vitro against HepG2, SGC7901, and HT29 through MTT assay. Moreover, the apoptosis-inducing activity of TSL-EO in SGC7901 cells was determined by Hoechst 33324 staining and flow cytometry methods. Also, the apoptosis-related proteins viz. Bax, Bcl-2 and caspase-3 were detected by western-blotting. Results GC–MS analysis showed that TSL-EO contained a high amount of sesquiterpenes (84.64%), including copaene (8.27%), β-caryophyllene (10.16%), caryophyllene (13.18%) and β-eudesmene (5.06%). TSL-EO inhibited the growth of both MSSA and MRSA, with the lowest MIC values of 0.125 and 1 mg/ml, respectively. Treatment with TSL-EO for 24 h could significantly suppress the viability of three different cancer cell lines (P, Graphical abstract

Purpose: To investigate the chemical composition of Bidens pilosa L. var. radiata . Sch Bip. (BP), as well as its antioxidant and anti-proliferative activities. Methods: The whole herb of BP was extracted with 95 % ethanol, which was then partitioned sequentially with petroleum ether, ethyl acetate and n-butyl alcohol to obtain petroleum ether, ethyl acetate (EE-BP), n-BuOH and water fractions. EE-BP was further isolated and purified by repeated column chromatography. The obtained compounds were identified by comparing 1H and 13C nuclear magnetic resonance (NMR) spectra with published data for the compounds. Flavonoids were evaluated for 1,1-diphenyl-2-picrylhydrazy (DPPH) and 2, 2 -azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) free radical scavenging activity by colorimetric methods, while their anti-proliferative effect against human colon cancer RKO cells was determined by 3-(4,5) dimethylthiahiazo (-z-y1)-3,5-diphenytetrazoliumromide (MTT) method. Results: A total of 6 compounds were isolated from BP, including flavonoids or flavonoid glycosides, among which were isoquercitrin (1), vitexin (2), astragalin (3), 5,6,7,4'-tetramethoxylflavone (4), 5,3',4'- trihydroxy-3,7-dimethoxylflavone (5) and quercetin (6). The results indicate that compound 6 was the most effective in both DPPH and ABTS free radical-scavenging assays, with half maximal inhibitory concentration (IC50) of 10.7 μmol/L and 17.5 μmol/L, respectively. Both compounds 4 and 5 exhibited significant growth inhibitory effect on RKO cells with IC50 of 39.08 μmol/L and 17.68 μmol/L, respectively (p < 0.01). Conclusion: BP contains a series of flavonoid compounds, possessing significant antioxidant activity and antiproliferative effect in RKO cells, thus suggesting that it is a potential health supplement and an easily available resource of natural antioxidants for use in colon cancer prevention. Keywords: Bidens pilosa , Flavonoid, Antioxidant, Anti-proliferation, Colon cancer

The in vitro antioxidant activity of ten different parts of lotus (Nelumbo nucifera Gaertn.) was evaluated and compared. Among the ten lotus extracts, the receptacle did not only possessed the highest phenolic, flavonoid and proanthocyanidin contents, but its 2-picrylhydrazyl free (DPPH) and 2, 2'-Azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, reducing power as well as total antioxidant activity were also comparable to, if not better than those of the butylated hydroxytoluene (BHT) control (p < 0.05). Compared with the BHT, all ten lotus extracts even exhibited significantly (p < 0.05) higher metal chelating activity. Nevertheless, the hydroxyl radical scavenging ability of all ten lotus extracts was significantly (p < 0.05) lower than that of the ascorbic acid control. In contrast to the metal chelating ability, phenolic compounds in the ten lotus extracts would likely be responsible for their DPPH and ABTS radical scavenging activities. Key words: Louts (Nelumbo nucifera), total phenolic content, total flavonoid content, total proanthocyanidin content, antioxidant activity.

The root of Morinda officinalis has been claimed to have a protective effect against bone loss in sciatic neurectomized and ovariectomized osteoporotic rats, and this protective effect is supposed to be attributed to anthraquinone compounds in the plant. In the present study, we investigated the effects of three anthraquinones isolated from M. officinalis, including 1, 3, 8-trihydroxy-2-methoxy-anthraquinone (1), 2-hydroxy-1-methoxy-anthraquinone (2) and rubiadin (3) on bone resorption activity in vitro and the mechanism on osteoclasts derived from rat bone marrow cells. Compound 1, 2 and 3 decreased the formation of bone resorption pits, the number of multinucleated osteoclasts, and the activity of tartrate resistant acid phosphates (TRAP) and cathepsin K in the coculture system of osteoblasts and bone marrow cells in the presence of 1, 25-dihydroxyvitamine D(3) and dexamethasone. They also enhanced the apoptosis of osteoclasts induced from bone marrow cells with M-CSF and RANKL. In addition, Compound 1, 2 and 3 improved the ratio of mRNA and protein expression of OPG and RANKL in osteoblasts, interfered with the JNK and NF-κB signal pathway, and reduced the expression of calcitonin receptor (CTR) and carbonic anhydrase/II (CA II) in osteoclasts induced from bone marrow cells with M-CSF and RANKL. These findings indicate that the anthraquinone compounds from M. officinalis are potential inhibitors of bone resorption, and may also serve as evidence to explain the mechanism of the inhibitory effects of some other reported anthraquinones on bone loss.

Eclipta prostrata, an aromatic plant, is known in Chinese herbal medicine for the treatment of various kidney diseases. In the present study, the volatile components were isolated from the aerial parts of this plant by hydrodistillation and analysed by GC-MS. A total of 55 compounds, which were the major part (91.7%) of the volatiles, were identified by matching mass spectra with a mass spectrum library (NIST 05.L). The main components were as follows: heptadecane (14.78%), 6,10,14-trimethyl-2-pentadecanone (12.80%), n-hexadecanoic acid (8.98%), pentadecane (8.68%), eudesma-4(14),11-diene (5.86%), phytol (3.77%), octadec-9-enoic acid (3.35%), 1,2-benzenedicarboxylic acid diisooctyl ester (2.74%), (Z,Z)-9,12-octadecadienoic acid (2.36%), (Z)-7,11-dimethyl-3-methylene-1,6,10-dodecatriene (2.08%) and (Z,Z,Z)-1,5,9,9-tetramethyl-1,4,7-cycloundecatriene (2.07%). The effects of volatile components and ethanolic extract from the aerial parts of this plant on the proliferation and differentiation of primary osteoblasts were evaluated by the MTT method and measuring the activity of alkaline phosphatase (ALP activity). Both volatile components and ethanolic extract (1 microg/mL to 100 microg/mL) significantly (p < 0.01) stimulated the proliferation and increased the ALP activity of primary osteoblasts. These results propose that E. prostrata can play an important role in osteoblastic bone formation, and may possibly lead to the development of bone-forming drugs.

Ganoderma, commonly referred to as “Lingzhi” in Chinese, “Reizhi” in Japanese, and “Youngzhi” in Korean, is a family of medicinal mushrooms employed to maintain human vitality and to promote longevity in traditional Chinese medicine for over 2000 years [1]. To date, Ganoderma has been used as remedy for the treatment of various diseases such as migraine and headache, hypertension, arthritis, hepatitis, cardiovascular problems, and cancer. It has been shown to contain a variety of bioactive ingredients including trierpenes, polysaccharides, nucleosides, steroids, fatty acids, alkaloids, proteins, peptides, amino acids, and inorganic elements [2]. Recently, Ganoderma spores gained considerable attention owning to their possible higher bioactivity [3]. However, spores have extremely hard and resilient bilayer walls, thus making them difficult to break open, and their bioactive lipid is stored between the inner and outer walls and is difficult to extract [4]. The difficulty above could be overcome by using superfine grinding technology through breaking the sporoderm and removing plant cellulose barriers [5]. Furthermore, it was reported that alcohol extracts from sporoderm-broken spores were more cytotoxic against tumor cells than that from sporoderm-nonbroken spores [6]. Super CO2 fluid extraction (SFE), a well-known extract technology, was established to extract liposoluble materials, which presented some advantages such as being nontoxic, high-efficiently, and less-deteriorating compared with the conventional methods viz. hydrodistillation, steam distillation, or solvent extraction [7]. In our current work, spores from Ganoderma duropora Lloyd (G. duropora) and Ganoderma lucidum (Leyss.ex Fr.) Karst. (G. lucidum) were superfine pulverized prior to extraction by SFE. The chemical composition of DSL (G. duropora spores lipids) and LSL (G. lucidum spores lipids) were then determined and compared by GC-MS. The results showed that spore lipids from G. duropora and G. lucidum obtained by SFE gave an average yield of 14.14% and 20.66% on a dry weight basis, respectively. As given in Table 1, GC-MS analyses revealed the presence of 22 and 10 compounds whose total relative contents were 74.04% and 88.58% in DSL and LSL, respectively. Aliphatic compounds were the main constituents in these two spore lipids, except for naphthalene and ergosterol belonging to aromatics and steroids, respectively. The chromatographic profile showed that both contained eight compounds and high levels of n-hexadecanoic acid and oleic acid (9.52%, 61.19% in DSL and 7.99%, 80.01% in LSL, as shown in Table 1). Nevertheless, 1-nonadecene (0.02%) and 2,3-dihydroxypropyl elaidate (0.07%) were detected in LSL but not in DSL. According to the above-mentioned results, it was concluded that the components from two spore lipids resembled each other. From this point of view, G. duropora in high production amounts might be developed as an alternative to Ganoderma.

Endophytic fungi possess favorable effects on their host plants, including disease-resistance improvement, secondary metabolite induction, and growth promotion. It is therefore a promising and sustainable strategy to utilize endophytic fungi for the quality improvement of medicinal herbs or important crops. In our study, a collection of 277 strains of endophytic fungi were isolated from Anoectochilus and Ludisia orchids. Two strains J162 and J211 can be symbiotically cocultured with the tissue culture seedlings of Anoectochilus roxburghii, a popular medicinal and edible plant in southern China. Both strains can significantly enhance the biomass of A. roxburghii and induce the biosynthesis and accumulation of its active ingredients, including flavonoids, kinsenoside, and polysaccharides. J162 and J211 were further identified as Chaetomium globosum and Colletotrichum gloeosporioides based on multilocus phylogenetic analysis. Immunocytochemical staining indicated that J162 and J211 mainly colonized the intercellular gap of xylem parenchyma cells of A. roxburghii roots without obvious harm. In addition, quantitative real-time polymerase chain reaction showed that the expression of three growth-related genes, namely, uracil phosphoribosyl transferase, amino acid transmembrane transporter, and maturase K, were significantly altered in A. roxburghii plants when treated with J162 and J211. In conclusion, the two strains are highly beneficial microbial resources for the growth and accumulation of active ingredients of A. roxburghii in agricultural cultivation.