Discovery of 6-Formylpyridyl Urea Derivatives as Potent Reversible-Covalent Fibroblast Growth Factor Receptor 4 Inhibitors with Improved Anti-Hepatocellular Carcinoma Activity.

Fibroblast growth factor receptor 4 (FGFR4) has been considered as a potential anticancer target due to FGF19/FGFR4 mediated aberrant signaling in hepatocellular carcinoma (HCC). Several FGFR4 inhibitors have been reported, but none have gained approval. Herein, a series of 5-formyl-pyrrolo[3,2-b]pyridine-3-carboxamides and a series of 6-formylpyridyl ureas were characterized as selective reversible-covalent FGFR4 inhibitors. The representative 6-formylpyridyl urea 8z exhibited excellent potency against FGFR4WT, FGFR4V550L, and FGFR4V550M with IC50 values of 16.3, 12.6, and 57.3 nM, respectively. It also potently suppressed proliferation of Ba/F3 cells driven by FGFR4WT, FGFR4V550L, and FGFR4V550M, and FGFR4-dependent Hep3B and Huh7 HCC cells, with IC50 values of 1.2, 13.5, 64.5, 15.0, and 20.4 nM, respectively. Furthermore, 8z displayed desirable microsomal stability and significant in vivo efficacy in the Huh7 HCC cancer xenograft model in nude mice. The study provides a promising new lead for anticancer drug discovery directed toward overcoming FGFR4 gatekeeper mutation mediated resistance in HCC patients.

Exploring the nurses' experiences in recognising and managing clinical deterioration in emergency patients: A qualitative study.

BACKGROUND: Emergency Department (ED) patients are particularly at a high risk of deterioration. The frontline nurses are key players in identifying and responding to deterioration events; however, few studies have sought to explore the whole process of recognition and management of clinical deterioration by emergency nurses. OBJECTIVES: The aim of this study was to explore the experiences of emergency nurses and provide a whole picture of how they recognise and manage clinical deterioration. METHODS: A qualitative descriptive study involving 11 senior nurses and seven junior nurses was conducted in the ED of a 3000-bed tertiary general hospital using semistructured interviews. The interviews were transcribed and thematically analysed. FINDINGS: Four salient themes emerged from the data analysis. The first, 'early recognition and response', revealed the importance of vital signs assessment in recognising and responding to clinical deterioration. The second, 'information transfer', depicted the skills and difficulties of transferring information in escalations of care. The third, 'abilities, education, and training', presented the abilities that emergency nurses should have and their perspectives on training. The fourth, 'support culture', described the major role of senior nurses in collaboration with colleagues in the ED. CONCLUSIONS: This study explored the experiences of emergency nurses in recognising and managing clinical deterioration. The findings illuminate the need to support the critical role of emergency nurses, with an emphasis on their abilities and continuous interprofessional collaboration training to improve the recognition and management of clinical deterioration.

Germacrane Sesquiterpene Dilactones from Mikania micrantha and Their Antibacterial and Cytotoxic Activity.

Four new germacrane sesquiterpene dilactones, 2ß-hydroxyl-11ß,13-dihydrodeoxymikanolide (1), 3ß-hydroxyl-11ß,13-dihydrodeoxymikanolide (2), 1α,3ß-dihydroxy-4,9-germacradiene-12,8:15,6-diolide (3), and (11ß,13-dihydrodeoxymikanolide-13-yl)-adenine (4), together with five known ones (5-9) were isolated from the aerial parts of Mikania micrantha. Their structures were elucidated on the basis of extensive spectroscopic analysis. Compound 4 is featured with an adenine moiety in the molecule, which is the first nitrogen-containing sesquiterpenoid so far isolated from this plant species. These compounds were evaluated for their in vitro antibacterial activity against four Gram-(+) bacteria of Staphyloccocus aureus (SA), methicillin-resistant Staphylococcus aureus (MRSA), Bacillus cereus (BC) and Curtobacterium. flaccumfaciens (CF), and three Gram-(-) bacteria of Escherichia coli (EC), Salmonella. typhimurium (SA), and Pseudomonas Solanacearum (PS). Compounds 4 and 7-9 were found to show strong in vitro antibacterial activity toward all the tested bacteria with the MIC values ranging from 1.56 to 12.5 µg/mL. Notably, compounds 4 and 9 showed significant antibacterial activity against the drug-resistant bacterium of MRSA with MIC value 6.25 µg/mL, which was close to reference compound vancomycin (MIC 3.125 µg/mL). Compounds 4 and 7-9 were further revealed to show in vitro cytotoxic activity toward human tumor A549, HepG2, MCF-7, and HeLa cell lines, with IC50 values ranging from 8.97 to 27.39 µM. No antibacterial and cytotoxic activity were displayed for the other compounds. The present research provided new data to support that M. micrantha is rich in structurally diverse bioactive compounds worthy of further development for pharmaceutical applications and for crop protection in agricultural fields.

Development and validation of an instrument for measuring junior nurses' recognition and response abilities to clinical deterioration (RRCD).

BACKGROUND: Nurses of all levels are expected to be competent in managing clinical deterioration. Given their limited experience and basic-level knowledge, there is a concern about junior nurses' clinical and patient management skills. However, junior nurses' abilities to recognise and respond to clinical deterioration have not been adequately explored because of the absence of a comprehensive tool. OBJECTIVES: The aim of this study was to develop a new self-assessment scale to assess the junior nurses' recognition and response abilities to clinical deterioration and to examine its reliability and validity. METHODS: Scale items were based on literature reviews and interviews. The preliminary scale was generated through two rounds of expert review. A panel of five experts evaluated content validity. After a pilot study, the questionnaire was distributed to 168 junior nurses via convenience sampling. Subsequent statistical analysis of results included construct validity, internal consistency, and test-retest reliability. RESULTS: Six factors were included, and 69.310% of the total variance was explained by the 25 items comprising the scale. The Cronbach's alpha coefficient was 0.905 (95% confidence interval [CI]: 0.812-0.979) for the overall scale and 0.655-0.838 for its subscales. The Guttman split-half reliability was 0.856 (95% CI: 0.806-0.894). The test-retest reliability of the scale was 0.878 (95% CI: 0.836-0.911). CONCLUSION: We developed a scale for measuring the abilities of junior nurses to recognise and respond to clinical deterioration and confirmed its reliability and validity. More experimental studies are needed to further evaluate this instrument.

Polycyclic Phenol Derivatives from the Leaves of Spermacoce latifolia and Their Antibacterial and α-Glucosidase Inhibitory Activity.

Three new polycyclic phenol derivatives, 2-acetyl-4-hydroxy-6H-furo [2,3-g]chromen-6-one (1), 2-(1',2'-dihydroxypropan-2'-yl)-4-hydroxy-6H-furo [2,3-g][1]benzopyran-6-one (2) and 3,8,10-trihydroxy-4,9-dimethoxy-6H-benzo[c]chromen-6-one (8), along with seven known ones (3-7, 9 and 10) were isolated for the first time from the leaves of Spermacoce latifolia. Their structures were determined by spectroscopic analysis and comparison with literature-reported data. These compounds were tested for their in vitro antibacterial activity against four Gram-(+) bacteria: Staphyloccocus aureus (SA), methicillin-resistant Staphylococcus aureus (MRSA), Bacillus cereus (BC), Bacillus subtilis (BS), and the Gram-(-) bacterium Escherichia coli. Compounds 1, 2, 5 and 8 showed antibacterial activity toward SA, BC and BS with MIC values ranging from 7.8 to 62.5 µg/mL, but they were inactive to MRSA. Compound 4 not only showed the best antibacterial activity against SA, BC and BS, but it further displayed significant antibacterial activity against MRSA (MIC 1.95 µg/mL) even stronger than vancomycin (MIC 3.9 µg/mL). No compounds showed inhibitory activity toward E. coli. Further bioassay indicated that compounds 1, 4, 5, 6, 8 and 9 showed in vitro α-glucosidase inhibitory activity, among which compound 9 displayed the best α-glucosidase inhibitory activity with IC50 value (0.026 mM) about 15-fold stronger than the reference compound acarbose (IC50 0.408 mM). These results suggested that compounds 4, 8 and 9 were potentially highly valuable compounds worthy of consideration to be further developed as an effective anti-MRSA agent or effective α-glucosidase inhibitors, respectively. In addition, the obtained data also supported that S. latifolia was rich in structurally diverse bioactive compounds worthy of further investigation, at least in searching for potential antibiotics and α-glucosidase inhibitors.

Comparison of the Performance of 24 Early Warning Scores with the Updated National Early Warning Score (NEWS2) for Predicting Unplanned Intensive Care Unit (ICU) Admission in Postoperative Brain Tumor Patients: A Retrospective Study at a Single Center.

BACKGROUND There have been few studies to evaluate early warning score (EWS) systems, or track and trigger systems (TTS), to identify early clinical deterioration in patients following brain tumor surgery who are admitted to the Intensive Care Unit (ICU). The National Early Warning Score (NEWS2) is an established method used in the U.K. National Health Service to improve care for in-hospital patients. This retrospective study from a single center aimed to compare the performance of NEWS2 with 24 other types of EWS to evaluate unplanned ICU admissions within 72 h after brain tumor surgery. MATERIAL AND METHODS A total of 326 patients with brain tumors were included in the study. Patients who experienced unplanned ICU transfer after surgery (69 cases) were diagnostically matched with patients who did not require intensive care (257 controls). We collected the physiological variables to calculate the area under the receiver operator characteristic curve (AUROC), sensitivity, specificity, Youden index values, cutoff values, positive predictive values, and negative predictive values. RESULTS The NEWS2 identified postoperative brain tumor patients with AUROC (0.860, p=0.000). The Patient-At-Risk (PAR) score was higher than NEWS2 in terms of AUROC value (0.870, P=0.000), Youden index (0.589 vs 0.542). CONCLUSIONS The findings showed that although the NEWS 2 performed well when used to evaluate unplanned ICU admissions within 72 h of postoperative brain tumor patients, the PAR score was also an accurate EWS.

Characterization of LEA genes in Dendrobium officinale and one Gene in induction of callus.

Late embryogenesis abundant (LEA) proteins are widely involved in plant stress responsive, while their involvement in callus formation is largest unknown. In this study, we identified and conducted expression analysis of the LEA genes from Phalaenopsis equestris and Dendrobium officinale, and characterized a LEA gene from D. officinale. A total 57 and 59 LEA genes were identified in P. equestris and D. officinale, respectively. A phylogenetic analysis showed that AtM, LEA_5 and Dehydrin groups were absent in both orchids. LEA_1 group genes were strongly expressed in seeds, significantly down-regulated in flowers, and absent in vegetative organs (leaves, stems and roots) in both orchids. Moreover, LEA_1 and LEA_4 group genes from D. officinale were abundant in the protocorm-like body stage and were dramatically up-regulated in response to abscisic acid and salinity stress. A LEA_1 gene (DoLEA43) was selected for further functional analysis. DoLEA43 protein was localized in the cytoplasm and nucleus, and its promoter contained a WUN-motif that was modulated by wounding. Overexpression of DoLEA43 in Arabidopsis enhanced callus induction, causing changes to callus formation-related genes such as WIND1. Our results indicate the involvement of LEA genes in the induction of callus, which provide insights into plant regeneration.

Thymol derivatives with antibacterial and cytotoxic activity from the aerial parts of Ageratina adenophora.

Three new thymol derivatives, 7-formyl-9-isobutyryloxy-8-hydroxythymol (1), 7,9-di-isobutyryloxy-8,10-dehydrothymol (2) and 2α-methoxyl-3ß-methyl-6-methylol-2,3-dihydrobenzofuran (3), along with five known ones (4-8), were isolated from the aerial parts of the invasive plant Ageratina adenophora. Their structures were elucidated by extensive spectroscopic analysis and they were all isolated from the aerial part of A. adenophora for the first time. These compounds, except 8, selectively showed in vitro antimicrobial activity against three Gram-(+) and two Gram-(-) bacterial strains. In particular, compounds 1 and 5 showed notable in vitro antimicrobial activity against all five bacterial strains with IC50 values ranging from 3.9 to 15.6 µg mL-1, as compared to reference compound kanamycin sulfate with a MIC value 1.9-3.9 µg mL-1. Compounds 1 and 5 were further revealed to show in vitro cytotoxic activity against three tested human tumor (MCF-7, NCI-H460 and HeLa) cell lines, with IC50 values ranging from 7.45 to 28.63 µM. Compounds 7 and 8 selectively showed slight but detectable in vitro cytotoxicity toward MCF-7 and NCI-H460 cell lines, with IC50 values 44.65-83.19 µM. No cytotoxic effects were detected in the bioassay of the other four thymol derivatives. The present results provide new data to support that the aerial parts of A. adenophora are a rich source of bioactive chemicals valuable in medicinal applications.

Development of the National Early Warning Score-Calcium Model for Predicting Adverse Outcomes in Patients With Acute Pancreatitis.

INTRODUCTION: This study aimed to develop a new model on the basis of the National Early Warning Score to predict intensive care unit admission and the mortality of patients with acute pancreatitis. METHODS: Patients diagnosed with acute pancreatitis in the emergency department were enrolled. The values of the National Early Warning Score, Modified Early Warning Score, and Bedside Index of Severity in Acute Pancreatitis in predicting intensive care unit admission and mortality of patients with acute pancreatitis were evaluated. RESULTS: A total of 379 patients with acute pancreatitis were enrolled; 77 patients (20.3%) were admitted to the intensive care unit and 14 (3.7%) died. The National Early Warning Score and calcium level were identified as independent risk factors of intensive care unit admission. Serum calcium exhibited a moderate correlation with National Early Warning Score (r = -0.46; P < 0.001), Modified Early Warning Score (r = -0.37; P < 0.001), and Bedside Index of Severity in Acute Pancreatitis (r = -0.39; P < 0.001). A new model called National Early Warning Score-calcium was developed by combining National Early Warning Score and calcium blood test result, which had larger areas under the curve for predicting intensive care unit admission and mortality than the other 3 scoring systems. DISCUSSION: A new model developed by combining National Early Warning Score and calcium exhibited better value in predicting the prognosis of acute pancreatitis than the models involving National Early Warning Score, Modified Early Warning Score, and Bedside Index of Severity in Acute Pancreatitis alone.

Mining MYB transcription factors from the genomes of orchids (Phalaenopsis and Dendrobium) and characterization of an orchid R2R3-MYB gene involved in water-soluble polysaccharide biosynthesis.

Members of the MYB superfamily act as regulators in a wide range of biological processes in plants. Despite this, the MYB superfamily from the Orchidaceae has not been identified, and MYB genes related to bioactive water-soluble polysaccharide (WSP) biosynthesis are relatively unknown. In this study, we identified 159 and 165 MYB genes from two orchids, Phalaenopsis equestris and Dendrobium officinale, respectively. The MYB proteins were classified into four MYB classes in both orchids: MYB-related (MYBR), R2R3-MYB, 3R-MYB and atypical MYB proteins. The MYBR proteins in both orchids were classified into five subfamilies and 12 genes were strongly up-regulated in response to cold stress in D. officinale. The R2R3-MYB proteins were both divided into 31 clades in P. equestris and D. officinale. Among these clades, nine contained MYB TFs related to secondary cell wall biosynthesis or testa mucilage biosynthesis in Arabidopsis thaliana. In D. officinale, 10 candidate genes showed an expression pattern corresponding to changes in the WSP content. Overexpression of one of these candidate genes (DoMYB75) in A. thaliana increased seed WSP content by about 14%. This study provides information about MYB genes in two orchids that will further help to understand the transcriptional regulation of WSP biosynthesis in these orchids as well as other plant species.

Diterpenoids with herbicidal and antifungal activities from hulls of rice (Oryza sativa).

Diterpenoids are the main secondary metabolites of plants and with a range of biological activities. In the present study, 7 compounds were isolated from the hulls of rice (Oryza sativa L.). Among them, 3 diterpenoids are new namely, 3,20-epoxy-3α-hydroxy- 8,11,13-abietatrie-7-one (1), 4,6-epoxy-3ß-hydroxy-9ß-pimara-7,15-diene (2) and 2-((E)-3- (4-hydroxy-3-methoxyphenyl) allylidene) momilactone A (3). While, 4 terpenoids are known, namely momilactone A (4), momilactone B (5), ent-7-oxo-kaur-15-en-18-oic acid (6) and orizaterpenoid (7). The structures of these diterpenoids were elucidated using 1D and 2D NMR in combination with ESI-MS and HR-EI-MS. Furthermore, all isolated compounds displayed antifungal activities against four crop pathogenic fungi Magnaporthe grisea, Rhizoctonia solani, Blumeria graminearum and Fusarium oxysporum, and phytotoxicity against paddy weed Echinochloa crusgalli. The results suggested that rice could produce plenty of secondary metabolites to defense against weeds and pathogens.

Anti-MRSA Sesquiterpenes from the Semi-Mangrove Plant Myoporum bontioides A. Gray.

The striking rise of methicillin-resistant Staphylococcus aureus (MRSA) infections has become a serious threat to public health worldwide. In an effort to search for new anti-MRSA agents from natural products, a bioassay-guided phytochemical study was conducted on the semi-mangrove plant Myoporum bontioides A. Gray, which led to the isolation of two new sesquiterpene alkaloids (1 and 2) and six known furanosesquiterpenes (3⁻8). Their structures were elucidated on the basis of extensive analysis of their 1D, 2D NMR and mass spectroscopic data. These two new alkaloids (1 and 2) displayed potent anti-MRSA activity with MIC value of 6.25 µg/mL. This is the first report of sesquiterpene alkaloids from the plants of Myoporum genus and their anti-MRSA activity.

Characterization and immunological activity of polysaccharides from Ixeris polycephala.

A water-soluble polysaccharide, named KMCP, was isolated and purified from edible plant Ixeris polycephala by using DEAE-52 cellulose chromatography. Its structure was determined by chemical analysis, methylation analysis, and NMR analysis, coupled with characterization by scanning electron spectroscopy (SEM). The resulting data indicated that KMCP was an arabinogalactan, with an average molecular weight of 1.95×106Da, which was mainly composed of arabinose and galactose in a relative molar ratio of 28.1% and 70.3%, respectively. The structure of KMPC was characterized as 72.5% of (1→4)-ß-Galp residues interspersed with 27.5% of (1→4,6)-ß-Galp residues in the main chain, and the branches were composed of (1→5)-α-Araf moieties or α-Araf (1→5) α-Araf (1→disaccharide moieties attached at O-6 of the (1→4,6)-ß-Galp residues. KMCP was revealed to be capable of exhibiting macrophage-mediated innate immune responses via enhancing phagocytosis of macrophages and increasing production of NO, activating NF-κB signaling pathway and promoting the mice spleen cells proliferation in a dose-dependent manner within the test concentrations (10.0-200.0µg/mL). These results suggested that KMCP could potentially be an effective and safe immunomodulator valuable to be utilized in pharmacological fields or in the development of functional foods.

Triterpenoids with α-glucosidase inhibitory activity and cytotoxic activity from the leaves of Akebia trifoliata.

Ten pentacyclic triterpenoids including a new multiflorane triterpene acid, 2α,3ß,23-trihydroxymultiflor-7-en-28-oic acid (1), and a new lupane triterpene monoglucoside named akebiaoside C (2), were obtained from the leaves of Akebia trifoliata. Their structures were elucidated by extensive spectroscopic analysis, and they were all isolated from the leaves of A. trifoliata for the first time. These compounds, except 4 and 5, showed in vitro α-glucosidase inhibitory activity much stronger than acarbose. Especially, 2, 3, 6, 8 and 10 displayed in vitro α-glucosidase inhibitory activity with IC50 values from 0.004 to 0.081 mM, which were close or even more potent than corosolic acid (IC50 0.06 mM). Triterpenoids 1, 8 and 10 were further revealed to show moderate in vitro cytotoxic activity against human tumor A549, HeLa and HepG2 cell lines, with IC50 values ranging from 26.5 to 51.9 µM. Compound 9 selectively showed in vitro cytotoxicity toward HeLa and HepG2 cell lines, with IC50 values of 81.49 and 73.47 µM, respectively. These findings provided new data to support that the leaves of A. trifoliata are a rich source in bioactive triterpenoids highly valuable to be developed for medicinal usage.

A Genome-Wide Identification of the WRKY Family Genes and a Survey of Potential WRKY Target Genes in Dendrobium officinale.

The WRKY family, one of the largest families of transcription factors, plays important roles in the regulation of various biological processes, including growth, development and stress responses in plants. In the present study, 63 DoWRKY genes were identified from the Dendrobium officinale genome. These were classified into groups I, II, III and a non-group, each with 14, 28, 10 and 11 members, respectively. ABA-responsive, sulfur-responsive and low temperature-responsive elements were identified in the 1-k upstream regulatory region of DoWRKY genes. Subsequently, the expression of the 63 DoWRKY genes under cold stress was assessed, and the expression profiles of a large number of these genes were regulated by low temperature in roots and stems. To further understand the regulatory mechanism of DoWRKY genes in biological processes, potential WRKY target genes were investigated. Among them, most stress-related genes contained multiple W-box elements in their promoters. In addition, the genes involved in polysaccharide synthesis and hydrolysis contained W-box elements in their 1-k upstream regulatory regions, suggesting that DoWRKY genes may play a role in polysaccharide metabolism. These results provide a basis for investigating the function of WRKY genes and help to understand the downstream regulation network in plants within the Orchidaceae.

Phenolics from Mikania micrantha and Their Antioxidant Activity.

A phytochemical study on the aerial parts of Mikania micrantha led to the isolation of two new phenolic compounds, benzyl 5-O-ß-d-glucopyranosyl-2,5-dihydroxybenzoate (1) and (7S,8R)-threo-dihydroxydehydrodiconiferyl alcohol 9-acetate (2), together with twelve known compounds, benzyl 2-O-ß-d-glucopyranosyl-2,6-dihydroxybenzoate (3), 4-allyl-2,6-dimethoxyphenol glucoside (4), (+)-isolariciresinol (5), icariol A2 (6), 9,10-dihydroxythymol (7), 8,9,10-trihydroxythymol (8), caffeic acid (9), p-coumaric acid (10), ethyl protocatechuate (11), procatechuic aldehyde (12), 4-hydroxybenzoic acid (13), and hydroquinone (14). Their structures were elucidated on the basis of extensive spectroscopic analysis. Except 8 and 9, all the other compounds were isolated from this plant species for the first time. The antioxidant activity of those isolated compounds were evaluated using three different assays. Compounds 1, 2, 3, 9, 10, 13, and 14 demonstrated significant 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) free radical cation scavenging activity ranging from SC50 0.31 to 4.86 µM, which were more potent than l-ascorbic acid (SC50 = 10.48 µM). Compounds 5, 9, 11, and 12 exhibited more potent 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (SC50 = 16.24-21.67 µM) than l-ascorbic acid (39.48 µM). Moreover, the ferric reducing antioxidant power (FRAP) of compounds 2, 5, 9, and 11 were discovered to be also comparable to or even more potent than l-ascorbic acid.

Two New Thymol Derivatives from the Roots of Ageratina adenophora.

Two new thymol derivatives, 7,9-diisobutyryloxy-8-ethoxythymol (1) and 7-acetoxy-8-methoxy-9-isobutyryloxythymol (2), were isolated from fresh roots of Ageratina adenophora, together with four known compounds, 7,9-di-isobutyryloxy-8-methoxythymol (3), 9-oxoageraphorone (4), (-)-isochaminic acid (5) and (1α,6α)-10-hydroxycar-3-ene-2-one (6). Their structures were established on the basis of detailed spectroscopic analysis, and they were all isolated from the roots of A. adenophora for the first time. All the compounds were tested for their in vitro antibacterial activity toward three Gram-positive and two Gram-negative bacterial strains. Thymol derivatives 1-3 only selectively showed slight in vitro bacteriostatic activity toward three Gram-positive bacteria. The two known carene-type monoterpenes 5 and 6 were found to show moderate in vitro antibacterial activity against all five tested bacterial strains, with MIC values from 15.6 to 62.5 µg/mL. In addition, compounds 5 and 6 were further revealed to show in vitro cytotoxicity against human tumor A549, HeLa and HepG2 cell lines, with IC50 values ranging from 18.36 to 41.87 µM. However, their cytotoxic activities were inferior to those of reference compound adriamycin.

Cytochemical Localization of Polysaccharides in Dendrobium officinale and the Involvement of DoCSLA6 in the Synthesis of Mannan Polysaccharides.

Dendrobium officinale is a precious traditional Chinese medicinal plant because of its abundant polysaccharides found in stems. We determined the composition of water-soluble polysaccharides and starch content in D. officinale stems. The extracted water-soluble polysaccharide content was as high as 35% (w/w). Analysis of the composition of monosaccharides showed that the water-soluble polysaccharides were dominated by mannose, to a lesser extent glucose, and a small amount of galactose, in a molar ratio of 223:48:1. Although starch was also found, its content was less than 10%. This result indicated that the major polysaccharides in D. officinale stems were non-starch polysaccharides, which might be mannan polysaccharides. The polysaccharides formed granules and were stored in plastids similar to starch grains, were localized in D. officinale stems by semi-thin and ultrathin sections. CELLULOSE SYNTHASE-LIKE A (CSLA) family members encode mannan synthases that catalyze the formation of mannan polysaccharides. To determine whether the CSLA gene from D. officinale was responsible for the synthesis of mannan polysaccharides, 35S:DoCSLA6 transgenic lines were generated and characterized. Our results suggest that the CSLA family genes from D. officinale play an important role in the biosynthesis of mannan polysaccharides.

A new anthraquinone and a new naphthoquinone from the whole plant of Spermacoce latifolia.

A phytochemical study on the whole plant of Spermacoce latifolia led to the isolation of a new anthraquinone, 1,2,6-trihydroxy-5-methoxy-9,10-anthraquinone (1), and a new naphthoquinone, (2R)-6-hydroxy-7-methoxy-dehydroiso-α-lapachone (2), together with three known anthraquinones (3-5). Their structures were established on the basis of detailed spectroscopic analysis, including one- and two-dimensional NMR, ESI-MS, and HR-ESI-MS techniques. All the compounds were isolated from S. latifolia for the first time. Compounds 1, 2, 4, and 5 showed significant antibacterial activity toward Bacillus subtilis with MIC values ranging from 0.9 to 31.2 µg/ml, and compound 4 aslo exhibited antibacterial activity against Bacillus cereus with a MIC value 62.5 µg/ml. Compound 1 was further revealed to show significant in vitro α-glucosidase inhibitory activity with IC50 value of 0.653 mM.

DoGMP1 from Dendrobium officinale contributes to mannose content of water-soluble polysaccharides and plays a role in salt stress response.

GDP-mannose pyrophosphorylase (GMP) catalyzed the formation of GDP-mannose, which serves as a donor for the biosynthesis of mannose-containing polysaccharides. In this study, three GMP genes from Dendrobium officinale (i.e., DoGMPs) were cloned and analyzed. The putative 1000 bp upstream regulatory region of these DoGMPs was isolated and cis-elements were identified, which indicates their possible role in responses to abiotic stresses. The DoGMP1 protein was shown to be localized in the cytoplasm. To further study the function of the DoGMP1 gene, 35S:DoGMP1 transgenic A. thaliana plants with an enhanced expression level of DoGMP1 were generated. Transgenic plants were indistinguishable from wild-type (WT) plants in tissue culture or in soil. However, the mannose content of the extracted water-soluble polysaccharides increased 67%, 96% and 92% in transgenic lines #1, #2 and #3, respectively more than WT levels. Germination percentage of seeds from transgenic lines was higher than WT seeds and the growth of seedlings from transgenic lines was better than WT seedlings under salinity stress (150 mM NaCl). Our results provide genetic evidence for the involvement of GMP genes in the biosynthesis of mannose-containing polysaccharides and the mediation of GMP genes in the response to salt stress during seed germination and seedling growth.