Hormonal and Metabolomic Responses of Dendrobium catenatum to Infection with the Southern Blight Pathogen Sclerotium delphinii.

Southern blight caused by Sclerotium delphinii has a devastating effect on Dendrobium catenatum (an extremely valuable medicinal and food homologous Orchidaceae plant). However, the mechanisms underlying S. delphinii infection and D. catenatum response are far from known. Here, we investigated the infection process and mode of S. delphinii through microscopic observations of detached leaves and living plantlets and further explored the hormonal and metabolomic responses of D. catenatum during S. delphinii infection by using the widely targeted metabolome method. The results showed that S. delphinii infection involves two stages: a contact phase (12 to 16 h after inoculation) and a penetration stage (20 h after inoculation). S. delphinii hyphae could penetrate leaves directly (via swollen hyphae and the formation of an infection cushion) or indirectly (via stomatal penetration), causing water-soaked lesions on leaves within 24 to 28 h after inoculation and expanded thereafter. The content of jasmonates increased after the hyphal contact and remained at high levels during S. delphinii infection, whereas the ethylene precursor (1-aminocyclopropanecarboxylic acid) accumulated significantly after penetration. Furthermore, metabolites of the phenylpropanoid and flavonoid pathways were enriched after pathogen penetration, whereas several amino acids accumulated in significant amounts at the late stage of infection. Moreover, some other associated metabolites were significantly altered during pathogen infection. Therefore, the jasmonate, phenylpropanoid, flavonoid, and amino acid pathways could play crucial roles in D. catenatum resistance to S. delphinii infection. This study provides insight into the prevention and control of southern blight disease of D. catenatum.

Predictive Nomogram for Midterm to Long-Term Prognosis in Patients with Papillary Renal Cell Carcinoma Based on Data from the Surveillance, Epidemiology, and End Results (SEER) Program.

BACKGROUND This study aimed to develop a predictive nomogram for midterm to long-term prognosis in patients with papillary renal cell carcinoma (RCC) based on data from the US Surveillance, Epidemiology, and End Results (SEER) program. MATERIAL AND METHODS Clinical pathology data and follow-up information were obtained from the SEER database for patients with papillary RCC between 1997-2014. Univariate and multivariate Cox regression models evaluated the independent prognostic factors, and the nomogram was constructed to predict the 3-year, 5-year, and 10-year survival rates. Multiple parameters were estimated to evaluate the predictive values, including the concordance indices (C-indices), calibration plots, area under the receiver operator characteristics (ROC) curve, net reclassification improvement (NRI), integrated discrimination improvement (IDI), and decision curve analysis (DCA). RESULTS The study included 13,926 patients with papillary RCC. Univariate and multivariate Cox regression analysis developed the nomogram that relied on the predictive variables of age, Fuhrman grade, TNM stage, surgery of the primary site, lymphadenectomy, and marital status. The C-indices of the novel model in the validation cohort were more satisfactory than those of the TNM classification. Accurate discrimination and calibration by the nomogram were identified in both cohorts. The NRI and IDI supported prediction improvements, and the DCA supported the nomogram's clinical significance. CONCLUSIONS A nomogram was developed to evaluate the prognosis of papillary RCC and to identify the patients who required specialized treatment. However, external validation of the predictive nomogram is required that also includes patients from other countries.

[Genome-wide identification and expression analysis of CSLA gene family of Dendrobium catenatum].

Glucomannan is the key active ingredient of Dendrobium catenatum, and CSLA family is responsible for glucomannan biosynthesis. In order to systematically evaluate the CSLA family members of D. catenatum, the bioinformatics methods were performed for genome-wide identification of DcCSLA gene family members through the genomic data of D. catenatum downloaded from the NCBI database, and further analyses of their phylogenetic relationship, gene structure, protein conserved domains and motifs, promoter cis-elements and gene expression profiles in response to stresses. The results showed that D. catenatum contains 13 CSLA members, all of which contain 9-10 exons. In the evolutionary relationship, CSLA genes were clustered into 5 groups, DcCSLA genes were distributed in all branches. Among which the ancestral genes of groupI existed before the monocot-dicot divergence, and groupⅡ-Ⅴ only existed in the monocot plants, indicating that group Ⅰ represents the earliest origin group. CSLA proteins are characteristic of the signature CESA_CaSu_A2 domain. Their promoter regions contain cis elements related to stresses and hormones. Under different stress treatments, low temperature induces the expression of DcCSLA5 and inhibits the expression of DcCSLA3. Infection of Sclerotium delphinii inhibits DcCSLA3/4/6/8/9/10 expression. Under the treatment of jasmonic acid, DcCSLA11 expression was significantly up-regulated, and DcCSLA2/5/7/12/13 were significantly down-regulated. These results laid a foundation for further study on the function of DcCSLA genes in glucomannan biosynthesis and accumulation.

A novel combination of quick response code and microfluidic paper-based analytical devices for rapid and quantitative detection.

Nowadays, there is great interest in the use of microfluidic paper-based analytical devices (µPADs) for low-cost diagnostics. In most cases, the test equipment is needed. Here we report a new type of device-independent detection method based on timer-paper-based analytical devices, which can be tested by smartphone, and its application for cholesterol detection. The Quick Response code was designed as the timer component of this device. Wax printing method was employed to print the pattern on filter paper. The color and enzyme reagents have been immobilized in the hydrophilic channel to complete the colorimetric detection for cholesterol. Under laminar flow conditions of the cellulose network, the liquid volume of detection zone has been quantified by monitoring the fluid residence time on different area of the timer-paper-based devices. The precise monitoring of detection time can promote the accuracy of colorimetric detection. This is very important for the quantitative detection of paper-based analytical devices. One significant outcome of this report is that simple and accurate timer can be used for detection process self-clocking. The factors of total detection time have been investigated. The linear range of the calibration curve was 3.0 ~ 6.0 mmol L-1, with correlation coefficient of 0.9956. With the characteristic of easy to use, low cost and accurate monitoring of detection time, this kind of timer-paper-based devices can be applied to cholesterol or other substances rapid detection.

DNA duplexes with hydrophobic modifications inhibit fusion between HIV-1 and cell membranes.

Discovery of new drugs for the treatment of AIDS typically possessing unique structures associated with novel mechanisms of action has been of great importance due to the quick drug-resistant mutations of HIV-1 strains. The work presented in this report describes a novel class of DNA duplex-based HIV-1 fusion inhibitors. Hydrophobic groups were introduced into a DNA duplex skeleton either at one end, at both ends, or in the middle. These modified DNA duplexes inhibited fusion between HIV-1 and human cell membranes at micro- or submicromolar concentrations. Respective inhibitors adopted an aptamer pattern instead of a base-pairing interaction pattern. Structure-activity relationship studies of the respective DNA duplexes showed that the rigid and negatively charged DNA skeletons, in addition to the presence of hydrophobic groups, were crucial to the anti-HIV-1 activity of these compounds. A fluorescent resonance energy transfer (FRET)-based inhibitory assay showed that these duplex inhibitors interacted with the primary pocket in the gp41 N-terminal heptad repeat (NHR) instead of interacting with the lipid bilayers.

Visual detection of copper(II) ions based on an anionic polythiophene derivative using click chemistry.

We have developed a novel approach for the rapid visual detection of Cu(2+) based on an anionic polythiophene derivative (sodium poly(2-(4-methyl-3-thienyloxy)propanesulfonate, PMTPS) using click chemistry. The method relies on the disassembly of PMTPS aggregates in the presence of cationic surfactant through electrostatic and hydrophobic interactions. In the assay of Cu(2+) detection, a cationic surfactant was formed via a click reaction catalyzed by copper(I), which was derived in situ from copper(II) and promoted the disassembly of PMTPS aggregates leading to the distinct solution color change from purple to yellow. This polymer probe has excellent sensitivity and selectivity for Cu(2+) with a detectable range in the micromolar regime by naked eyes and can be used for monitoring Cu(2+) concentrations below the safety limit in real-world samples.

Colorimetric and fluorescent dual detection of paraquat and diquat based on an anionic polythiophene derivative.

We have developed a colorimetric and fluorescent dual-response probe for the detection of paraquat and diquat in aqueous solutions based on an anionic polythiophene derivative. The detection limit of this approach can be as low as 10(-9) M by fluorescence measurements.

Colorimetric and fluorescent detection of protamines with an anionic polythiophene derivative.

We have developed a novel approach for the colorimetric and fluorescent detection of protamines based on an anionic polythiophene derivative. This probe has high sensitivity and selectivity toward protamines with a linear detectable range from 0.1 to 30 µg mL(-1), and can be used for protamine detection in serum samples.

Interactions between endophytic fungus Pestalotiopsis sp. DO14 and Dendrobium catenatum: Deciphering plant polysaccharide and flavonoid accumulation and underlying mechanisms by comparative transcriptome and metabolome analyses.

Dendrobium catenatum, which belongs to the Orchidaceae family, has been used as a traditional medicine and healthy food in China for over 2000 years, and is of enormous economic value. Polysaccharides and flavonoids are two major functional ingredients in D. catenatum stems that contribute to its health benefits. D. catenatum lives in close association with endophytic fungi, but the literature regarding the further relations between them, especially the fungal-induced accumulation of metabolites in the host plant, is sparse. Our previous study showed that Pestalotiopsis sp. DO14 isolated from D. catenatum improved the host plant growth and metabolite accumulation. This study was performed to investigate dynamic variations of the growth traits, key metabolites (polysaccharides and flavonoids), and expression of key genes of D. catenatum under conditions of the DO14 colonization. Colonization with DO14 promoted D. catenatum growth as indicated by increased leaf area, mid-stem thickness, and plant height. The content of polysaccharides, mannose, and sucrose increased even without DO14 entering the host cells or forming a mature symbiotic relationship concurrent with improved photosynthesis rate. Furthermore, DO14 induced upregulation of genes involved in sugar and flavonoid metabolism, especially phosphoenolpyruvate carboxykinase (PCKA), chalcone synthase (CHS) and UDP-glycose flavonoid glycosyltransferase (UFGT). These observations suggested that endophytic fungi induce the accumulation of polysaccharides and flavonoids by plants, increasing the efficiency of carbon assimilation and carbon turnover. The findings of this study provide insight into the mechanisms underlying Orchidaceae-endophyte interactions, and suggest potential novel applications of endophytic fungi in D. catenatum breeding to improved plant quality.

Machine learning uncovers accumulation mechanism of flavonoid compounds in Polygonatum cyrtonema Hua.

The compositions and yield of flavonoid compounds of Polygonatum cyrtonema Hua (PC) are important indices of the quality of medicinal materials. However, the flavonoids compositions and accumulation mechanism are still unclear in PC. Here, we identified 22 flavonoids using widely-targeted metabolome analysis in 15 genotypes of PC. Then weighted gene co-expression network analysis based on 45 transcriptome samples was performed to construct 12 co-expressed modules, in which blue module highly correlated with flavonoids was identified. Furthermore, 4 feature genes including PcCHS1, PcCHI, PcCHS2 and PcCHR5 were identified from 94 hub genes in blue module via machine learning methods support vector machine-recursive feature elimination (SVM-RFE) and random forest (RF), and their functions on metabolic flux of flavonoids pathway were confirmed by tobacco transient expression system. Our findings identified representative flavonoids and key enzymes in PC that provided new insight for elite breeding rich in flavonoids, and thus will be beneficial for rapid development of great potential economic and medicinal value of PC.

Reversion of p-glycoprotein-mediated multidrug resistance in human leukemic cell line by diallyl trisulfide.

Multidrug resistance (MDR) is the major obstacle in chemotherapy, which involves multiple signaling pathways. Diallyl trisulfide (DATS) is the main sulfuric compound in garlic. In the present study, we aimed to explore whether DATS could overcome P-glycoprotein-(P-gp-)mediated MDR in K562/A02 cells, and to investigate whether NF-κB suppression is involved in DATS-induced reversal of MDR. MTT assay revealed that cotreatment with DATS increased the response of K562/A02 cells to adriamycin (the resistance reversal fold was 3.79) without toxic side effects. DATS could enhance the intracellular concentration of adriamycin by inhibiting the function and expression of P-gp, as shown by flow cytometry, RT-PCR, and western blot. In addition, DATS resulted in more K562/A02 cell apoptosis, accompanied by increased expression of caspase-3. The expression of NF-κB/p65 (downregulation) was significantly linked to the drug-resistance mechanism of DATS, whereas the expression of IκBα was not affected by DATS. Our findings demonstrated that DATS can serve as a novel, nontoxic modulator of MDR, and can reverse the MDR of K562/A02 cells in vitro by increasing intracellular adriamycin concentration and inducing apoptosis. More importantly, we proved for the first time that the suppression of NF-κB possibly involves the molecular mechanism in the course of reversion by DATS.

Design and evaluation of a novel evodiamine-phospholipid complex for improved oral bioavailability.

A novel evodiamine (EVO)-phospholipid complex (EPLC) was designed to improve the bioavailability of EVO. A central composite design approach was employed for process optimization. EPLC were characterized by differential scanning calorimetry, ultraviolet spectroscopy, Fourier transformed infrared spectroscopy, (1)H-NMR spectroscopy, matrix-assisted laser desorption/ionization time-of-flight spectroscopy, apparent solubility, and dissolution rate. After oral administration of EPLC, the concentrations of EVO at different time points were determined by high-performance liquid chromatography. The optimal formulation for EPLC was obtained where the values of X (1), X (2), and X (3) were 2, 0.5, and 2.5 mg/mL, respectively. The average particle size and zeta potential of EPLC with the optimized formulation were 246.1 nm and -26.94 mV, respectively. The EVO and phospholipids in the EPLC were associated with non-covalent interactions. The solubility of EPLC in water and the dissolution rate of EPLC in phosphate-buffered solution (pH 6.8) were substantially enhanced. The plasma EVO concentration-time curves of EPLC and free EVO were both in accordance with the two-compartment model. The peak concentration and AUC(0-∞) of EPLC were increased, and the relative bioavailability was significantly increased to 218.82 % compared with that of EVO.

Genome-wide analysis of basic helix-loop-helix genes in Dendrobium catenatum and functional characterization of DcMYC2 in jasmonate-mediated immunity to Sclerotium delphinii.

Dendrobium catenatum, belonging to the Orchidaceae, is a precious Chinese herbal medicine. Sclerotium delphinii (P1) is a broad-spectrum fungal disease, which causes widespread loss in the near-wild cultivation of D. catenatum. Thus, resistance breeding of D. catenatum has become the key to solve this problem. The basic helix-loop-helix (bHLH) gene family is closely related to plant resistance to external stresses, but the related research in D. catenatum is not deep enough yet. Phylogenetic analysis showed that 108 DcbHLH genes could be divided into 23 subgroups. Promoter cis-acting elements revealed that DcbHLHs contain a large number of stress-related cis-acting elements. Transcriptome analysis of MeJA and P1 treatment manifested that exogenous MeJA can change the expression pattern of most bHLH genes, especially the IIIe subgroup, including inhibiting the expression of DcbHLH026 (MYC2a) and promoting the expression of DcbHLH027 (MYC2b). Subcellular localization indicated that they were located in the nucleus. Furthermore, exogenous MeJA treatment significantly delayed disease time and reduced lesion size after infection with P1. DcMYC2b-overexpression Arabidopsis lines showed significantly smaller lesions after being infected with P1 than the wild type, indicating that DcMYC2b functions as an important positive regulator in D. catenatum defense against P1. Our findings shed more insights into the critical role of the DcbHLH family in plants and the resistance breeding of D. catenatum.

Pathways Linking Parental Support to Adolescents' Reading Proficiency: A Social Cognitive Theory Perspective.

Parental support is essential to children's motivation and academic functioning. However, few studies have investigated the pathways linking perceived parental support to children's achievement in reading during adolescence. This study aims to fill this gap by systematically investigating the relationships among perceived support from parents, adolescents' motivational beliefs, and reading proficiency based on Bandura's social cognitive theory. A range of motivational processes are explored, including self-efficacy, goals, and values. Using the China sample from the Programme for International Student Assessment (PISA) 2018, which includes 12,058 adolescents from 361 schools, this study proposed two competing models based on different accounts of self-efficacy beliefs. Multilevel path analysis is adopted as the analytic method. The results suggest that perceived support from parents has a statistically significant but negligible relationship with adolescents' reading proficiency. However, this relationship is mediated by nuanced pathways such as self-efficacy beliefs, mastery goal orientation, and reading enjoyment. Findings of this study provide evidence in support of the top-down theory of self-efficacy in the reading context and also contribute to a better understanding of the interactions between different motivational processes. Theoretical and practical implications of this study are discussed, and suggestions for future research are offered.

Differentiating the learning styles of college students in different disciplines in a college English blended learning setting.

Learning styles are critical to educational psychology, especially when investigating various contextual factors that interact with individual learning styles. Drawing upon Biglan's taxonomy of academic tribes, this study systematically analyzed the learning styles of 790 sophomores in a blended learning course with 46 specializations using a novel machine learning algorithm called the support vector machine (SVM). Moreover, an SVM-based recursive feature elimination (SVM-RFE) technique was integrated to identify the differential features among distinct disciplines. The findings of this study shed light on the optimal feature sets that collectively determined students' discipline-specific learning styles in a college blended learning setting.

Transcriptomic and Metabolomic Approaches Deepen Our Knowledge of Plant-Endophyte Interactions.

In natural systems, plant-symbiont-pathogen interactions play important roles in mitigating abiotic and biotic stresses in plants. Symbionts have their own special recognition ways, but they may share some similar characteristics with pathogens based on studies of model microbes and plants. Multi-omics technologies could be applied to study plant-microbe interactions, especially plant-endophyte interactions. Endophytes are naturally occurring microbes that inhabit plants, but do not cause apparent symptoms in them, and arise as an advantageous source of novel metabolites, agriculturally important promoters, and stress resisters in their host plants. Although biochemical, physiological, and molecular investigations have demonstrated that endophytes confer benefits to their hosts, especially in terms of promoting plant growth, increasing metabolic capabilities, and enhancing stress resistance, plant-endophyte interactions consist of complex mechanisms between the two symbionts. Further knowledge of these mechanisms may be gained by adopting a multi-omics approach. The involved interaction, which can range from colonization to protection against adverse conditions, has been investigated by transcriptomics and metabolomics. This review aims to provide effective means and ways of applying multi-omics studies to solve the current problems in the characterization of plant-microbe interactions, involving recognition and colonization. The obtained results should be useful for identifying the key determinants in such interactions and would also provide a timely theoretical and material basis for the study of interaction mechanisms and their applications.

Antecedents of adolescent students' ICT self-efficacy: The ICT dataset.

Based on the Programme for International Student Assessment (PISA) 2015 dataset, the information and communication technology (ICT) dataset focuses specifically on ICT-related constructs in the context of educational technology. It includes a wide range of student-level variables collected from 30 Economic Co-operation and Development (OECD) countries, which pertain to students' motivational and behavioural characteristics in relation to their ICT self-efficacy. In total, it comprises 201, 652 students from 7708 schools. As technology has become an integral component of education, the ICT dataset can serve as a handy resource for studying ICT-related constructs. Besides, the ICT dataset holds advantages over the original PISA dataset for its intensive focus and easy readability. With this important resource, researchers can undertake their own research in the neighbouring fields of ICT, developing their own theories or validating existing theoretical frameworks and statements. The focus of this study is to identify the antecedents of adolescent students' ICT self-efficacy and illuminate potential mechanisms at work.

NEK7: a potential therapy target for NLRP3-related diseases.

NLRP3 inflammasome plays an essential role in innate immunity, yet the activation mechanism of NLRP3 inflammasome is not clear. In human or animal models, inappropriate NLRP3 inflammasome activation is implicated in many NLRP3-related diseases, such as tumors, inflammatory diseases and autoimmune diseases. Until now, a great number of inhibitors have been used to disturb the related signaling pathways, such as IL-1ß blockade, IL-18 blockade and caspase-1 inhibitors. Unfortunately, most of these inhibitors just disturb the signaling pathways after the activation of NLRP3 inflammasome. Inhibitors that directly regulate NLRP3 to abolish the inflammation response may be more effective. NEK7 is a multifunctional kinase affecting centrosome duplication, mitochondrial regulation, intracellular protein transport, DNA repair and mitotic spindle assembly. Researchers have made significant observations on the regulation of gene transcription or protein expression of the NLRP3 inflammasome signaling pathway by NEK7. Those signaling pathways include ROS signaling, potassium efflux, lysosomal destabilization, and NF-κB signaling. Furthermore, NEK7 has been proved to be involved in many NLRP3-related diseases in humans or in animal models. Inhibitors focused on NEK7 may regulate NLRP3 to abolish the inflammation response and NEK7 may be a potential therapeutic target for NLRP3-related diseases.

NEK7 interacts with NLRP3 to modulate the pyroptosis in inflammatory bowel disease via NF-κB signaling.

Inflammatory bowel disease (IBD) is one of the most common diseases in the gastrointestinal tract related to aberrant inflammation. Pyroptosis, which is characterized by inflammasome formation, the activation of caspase-1, and the separation of the N- and C-terminals of GSDMD, might be related to IBD pathogenesis. NEK7 is an important component of the NLRP3 inflammasome in macrophages. We attempted to investigate the mechanism of NEK7 interacting with NLRP3 to modulate the pyroptosis in IBD. NEK7 mRNA and protein expression and pyroptosis-associated factors, including Caspase-1 (p45, p20), NLRP3, and GSDMD, were upregulated in IBD tissues. NEK7 knockdown abolish ATP + LPS-induced pyroptosis in vitro and improved DSS-induced chronic colitis in vivo. NEK7 interacted with NLRP3, as revealed by Co-IP and GST pull-down assays, to exert its effects. Moreover, short-term LPS treatment alone induced no significant changes in NEK7 protein level. TLR4/NF-κB signaling in MODE-K cells could be activated by LPS treatment. LPS-induced NEK7 upregulation could be significantly reversed by JSH-23, an inhibitor of p65. Furthermore, LUC and ChIP assays revealed that RELA might activate the transcription of NEK7 via targeting its promoter region. LPS-induced TLR4/NF-κB activation causes an increase in NEK7 expression by RELA binding NEK7 promoter region. In conclusion, NEK7 interacts with NLRP3 to modulate NLRP3 inflammasome activation, therefore modulating the pyroptosis in MODE-K cells and DSS-induced chronic colitis in mice. We provide a novel mechanism of NEK7-NLRP3 interaction affecting IBD via pyroptosis.

Expression of FXR and HRG and their clinicopathological significance in benign and malignant pancreatic lesions.

AIMS: The following study examines the FXR and HRG expression in benign and malignant lesions of the pancreas and evaluates the association between FXR and HRG expression with clinicopathological features and prognosis of pancreatic cancer. MATERIALS AND METHODS: Immunohistochemistry of FXR and HRG was performed with EnVision™ in 106 pancreatic ductal adenocarcinoma (PDAC) specimens, 35 paracancer samples (2 cm away from the tumor, when possible or available), 55 benign lesions and 13 normal tissue samples. RESULTS: The percentage of cases with positive FXR and negative HRG expression was significantly higher in PDAC compared to pericancerous tissues, benign lesions and normal tissues (P<0.05 or P<0.01). In pancreatic tissues with benign lesions, tissues with positive FXR and/or negative HRG protein expression exhibited dysplasia or intraepithelial neoplasia. The percentage of cases with positive FXR and negative HRG expressions was significantly higher in PDAC with lymph node metastasis, invasion, and TNM stage III+IV disease (P<0.05 or P<0.01). The expression of FXR was negatively correlated with HRG (P<0.05). In addition, the univariate Kaplan-Meier analysis showed that positive FXR and negative HRG expression, poor differentiation, large tumor size, high TNM stage, lymph node metastasis, and invasion were closely associated with decreased overall survival in PDAC patients (P<0.05 or P<0.01). Moreover, multivariate Cox regression analysis identified that positive FXR and negative HRG expression were independent factors for poor prognosis in PDAC. The AUC for FXR was (AUC=0.709, 95% CI: 0.632-0.787), and for HRG was (AUC=0.719, 95% CI: 0.643-0.796) in PDAC compared to benign lesions. CONCLUSIONS: Positive FXR and negative HRG expression are closely associated with the carcinogenesis, clinical, pathological and biological behaviors, and poor prognosis in PDAC.