Small RNA sequencing provides insights into molecular mechanism of flower development in Rhododendron pulchrum Sweet.

Rhododendron pulchrum sweet, a member of the Ericaceae family possessing valuable horticultural properties, is widely distributed in the temperate regions. Though serving as bioindicator of metal pollution, the molecular mechanism regulating flowering in R. pulchrum is very limited. Illumina sequencing was performed to identify critical miRNAs in the synthesis of flavonoids at different developmental stages. Totally, 722 miRNAs belonging to 104 families were screened, and 84 novel mature miRNA sequences were predicted. The miR166, miR156, and miR167-1 families were dominant. In particular, 126 miRNAs were significantly differentially expressed among four different flowering stages. Totally, 593 genes were differentially regulated by miRNAs during the flower development process, which were mostly involved in "metabolic pathways", "plant hormone signal transduction", and "mitosis and regulation of biosynthetic processes". In pigment biosynthesis and signal transduction processes, gra-miR750 significantly regulated the expression of flavonoid 3',5'-hydroxylase; aof-miR171a, aof-miR171b, aof-miR171c, cas-miR171a-3p, and cas-miR171c-3p could regulate the expression of DELLA protein; aof-miR390, aof-miR396b, ath-miR3932b-5p, cas-miR171a-3p, aof-miR171a, and aof-miR171b regulated BAK1 expression. This research showed great potentials for genetic improvement of flower color traits for R. pulchrum and other Rhododendron species.

Growth differentiation factor 15 predicts cardiovascular events in stable coronary artery disease.

BACKGROUND: Growth differentiation factor 15 (GDF-15) has been explored as a potential biomarker for various inflammatory diseases and cardiovascular events. This study aimed to assess the predictive role of GDF-15 levels in cardiovascular events and all-cause mortality, considering traditional risk factors and other biomarkers. METHODS: A prospective study was conducted and 3699 patients with stable coronary artery disease (CAD) were enrolled into the research. Baseline GDF-15 levels were measured. Median follow-up was 3.1 years during the study. We analyzed clinical variables and several biomarkers. Multivariable Cox regression analysis was performed to evaluate prognostic performance of GDF-15 levels in predicting myocardial infarction (MI), heart failure, stroke, cardiovascular death, and non-cardiovascular death. RESULTS: Baseline GDF-15 levels for 3699 patients were grouped by quartile (≤ 1153, 1153-1888, 1888-3043, > 3043 ng/L). Higher GDF-15 levels were associated with older age, male gender, history of hypertension, and elevated levels of N-terminal pro B-type natriuretic peptide (NT-pro BNP), soluble suppression of tumorigenesis-2 (sST2), and creatine (each with P < 0.001). Adjusting for established risk factors and biomarkers in Cox proportional hazards models, a 1 standard deviation (SD) increase in GDF-15 was associated with elevated risk of clinical events [hazard ratio (HR) = 2.18, 95% confidence interval (CI): (1.52-3.11)], including: MI [HR = 2.83 95% CI: (1.03-7.74)], heart failure [HR = 2.71 95% CI: (1.18-6.23)], cardiovascular and non-cardiovascular death [HR = 2.48, 95% CI (1.49-4.11)] during the median follow up of 3.1 years. CONCLUSIONS: Higher levels of GDF-15 consistently provides prognostic information for cardiovascular events and all cause death, independent of clinical risk factors and other biomarkers. GDF-15 could be considered as a valuable addition to future risk prediction model in secondary prevention for predicting clinical events in patient with stable CAD.

Insights into the contributions of hemicelluloses to assembly and mechanical properties of cellulose networks.

In order to explore the contributions of hemicelluloses to the assembly and mechanical properties of cellulose networks, the bacterial cellulose (BC) composites containing xylan and glucomannan were prepared to mimic the polysaccharides network of plant cell walls. Both polysaccharides could induce the change of diameters of cellulose ribbons and influence the crystallization of cellulose. Besides, small-angle X-ray scattering (SAXS) demonstrated that xylan inhibited the assembly of microfibrils into cellulose ribbons, while glucomannan promoted the packing of microfibrils. The changes of cellulose crystalline structure and assembly pattern of cellulose contributed to the lower tensile strength and higher strain at break of the BC composites as compared with the BC. The results provide a profound insight into the structure-property relationships of cellulose networks affected by hemicelluloses, which could be conducive to the development of cellulose biomaterials.

Mild pretreatment with Brønsted acidic deep eutectic solvents for fractionating ß-O-4 linkage-rich lignin with high sunscreen performance and evaluation of enzymatic saccharification synergism.

Herein, a mild fractionation method by employing polyol-based Brønsted acidic DESs (BDESs) was proposed to extract lignin with well-preserved ß-O-4 substructures and to enhance fermentable sugar yields simultaneously. For ethylene glycol-oxalic acid (EG-OA), more than 53 % of lignin was obtained and superb carbohydrate digestibility (i.e., glucose and xylose yields were reached to 94.6 % and 87.7 %, respectively) was achieved after pretreatment. Remarkably, detailed structural studies revealed that the polyol was incorporated into lignin, which stabilized reactive carbocation intermediates formed during BDESs treatment and prevented undesired recondensation reactions. This lignin protection mechanism was shown to play a key role in enzymatic hydrolysis enhancement and lignin valorization. The resultant ß-O-4 linkage-rich lignin fractions were attractive for natural sunscreen applications due to their lighter color and excellent UV-blocking performance. Overall, this work proposed a sustainable and economically practical lignin-first biorefinery approach that is beneficial for achieving comprehensive valorization of lignocellulose.

Multifunctional hybrid hydrogel with transparency, conductivity, and self-adhesion for soft sensors using hemicellulose-decorated polypyrrole as a conductive matrix.

Polymers with high conductivity and cross-linking ability are ideal materials for the preparation of conductive hydrogels for application in wearable electronic devices. However, the fabrication of conductive polymer-incorporated hydrogels with good synergistic properties remains a great challenge due to the hydrophobicity and opacity of conjugated π conductive polymers. In this study, a multifunctional hybrid hydrogel was prepared by incorporating hemicellulose-decorated polypyrrole (H/PPY), polyvinyl alcohol (PVA), and tannic acid (TA) into a polyacrylamide (PAM) network. The addition of excess ammonium persulfate (APS) in the process of gelation not only initiated the polymerization of PAM but also resulted in the change of the hydrogel from opaque to transparent by continuously breaking and reducing the size of the PPY particles. The hybrid hydrogel exhibited high transparency and conductivity, good adhesion ability and mechanical performance, and high resistance strain sensitivity and could accurately monitor the strain signals of the index finger and elbow flexion and pulse beat during rest and exercise, which has promising potential for use in wearable or implantable smart sensor devices, electronic skins, and artificial intelligence applications.

Prediction of SYNTAX score II improvement by adding temporal heart rate changes between discharge and first outpatient visit in patients with acute myocardial infarction.

BACKGROUND: The prognostic ability of the temporal changes in resting heart rate (ΔHR) in patients with acute myocardial infarction (AMI) for cardiovascular (CV) mortality and clinical outcomes is rarely examined. This study investigated the predictive value of ΔHR using models with SYNTAX score II (SxS-II) for the long-term prognosis of patients with AMI. METHODS: Six hundred five AMI patients with vital signs recorded at the first outpatient visit (2-4 weeks after discharge) were retrospectively recruited into this study. The changes between discharge and outpatient resting heart rate (D-O ΔHR) were calculated by subtracting the HR at the first post-discharge visit from the value recorded at discharge. The major adverse cardiovascular and cerebrovascular events (MACCE) include cardiovascular death, recurrent myocardial infarction, revascularization, and nonfatal stroke. The predictive values and reclassification ability of the different models were assessed using a likelihood ratio test, Akaike's information criteria (AIC), receiver operating characteristic (ROC) curves, net reclassification improvement (NRI), and integrated discrimination improvement (IDI). RESULTS: During the follow-up period, a drop-in resting heart rate (RHR) from discharge to first outpatient visit was independently associated with less risk of CV mortality [D-O ΔHR: hazards ratio (HR) = 0.97, 95% CI = 0.96-0.99, P < 0.001] and MACCE (HR = 0.98, 95% CI = 0.97-0.99, p = 0.001). The likelihood test indicated that the combined model of SxS-II and D-O ΔHR yielded the lowest AIC for CV mortality and MACCE (P < 0.001). Moreover, D-O ΔHR alone significantly improved the net reclassification and integrated discrimination of the models containing SxS-II for CV mortality and MACCE (CV mortality: NRI = 0.5600, P = 0.001 and IDI = 0.0759, P = 0.03; MACCE: NRI = 0.2231, P < 0.05 and IDI = 0.0107, P < 0.05). CONCLUSIONS: The change in D-O ΔHR was an independent predictor of long-term CV mortality and MACCE. The D-O ΔHR combined with SxS-II could significantly improve its predictive probability.

Visual and Rapid Detection of Porcine Epidemic Diarrhea Virus (PEDV) Using Reverse Transcription Loop-Mediated Isothermal Amplification Method.

Porcine epidemic diarrhea virus (PEDV) can cause severe infectious porcine epidemic diarrhea (PED) and infect different ages of pigs, resulting in sickness and death among suckling pigs. For PEDV detection, finding an effective and rapid method is a priority. In this study, we established an effective reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for PEDV detection. Three sets of primers, specific for eight different sequences of the PEDV N gene, were designed in this study. The optimized RT-LAMP amplification program was as follows: 59 min at 61.9 °C and 3 min at 80 °C. The RT-LAMP results were confirmed with the addition of SYBR Green I fluorescence dye and with the detection of a ladder-like band by conventional gel electrophoresis analysis, which demonstrated a significant agreement between the two methods. The LOD of PEDV by RT-LAMP was 0.0001 ng/µL. Compared with RT-LAMP, the traditional RT-PCR method is 100-fold less sensitive. The RT-LAMP results had no cross-reaction with porcine parvovirus (PPV), porcine circovirus type 1 (PCV1), porcine pseudorabies virus (PRV), porcine circovirus type 2 (PCV2), rotavirus (RV), transmissible gastroenteritis virus (TGEV) and porcine reproductive and respiratory syndrome virus (PRRSV). Consequently, the newly developed RT-LAMP method could provide an accurate and reliable tool for PEDV diagnosis.

Research progress of signaling pathways of the natural substances intervene dyslipidemia (Review).

Dyslipidemia is an umbrella term for a range of lipid metabolic disorders in the body. This condition has been widely reported to greatly increase the risk of cardiovascular diseases, threatening human health. In recent years, advances in molecular biology have deepened understanding of the dyslipidemia-related signaling pathways and specific mechanisms underlying dyslipidemia. Signaling pathways possess the ability to transmit an extracellular signal to the inside of the cell, leading to specific biological effects. Lipid metabolism disorders and lipid levels in the blood are frequently affected by aberrant alterations in the dyslipidemia-related signaling pathways. Therefore, further investigations into these pathways are required for the prevention and treatment of dyslipidemia. The present review summarizes the characteristics of six dyslipidemia-associated signaling pathways: Peroxisome proliferator-activated receptor, adenosine monophosphate-activated protein kinase, farnesoid X receptor, forkhead box O, adipocytokine and cyclic adenosine monophosphate signaling pathways. In particular, specific focus was placed on previous experimental studies and reports on the intervention effects of natural substances (compounds from animals, plants, marine organisms and microorganisms) on dyslipidemia.

Association between Gut Microbiota Dysbiosis and the CHA2DS2-VASc Score in Atrial Fibrillation Patients.

Background: In our previous studies, we found a disordered taxonomic composition and function of gut microbiota (GM) in atrial fibrillation (AF) patients. However, direct evidence about the association between dysbiotic microbiota and thromboembolic risk in AF is lacking. Aims: In this study, we analyzed the interaction of GM and related functional patterns in AF with different CHA2DS2-VASc scores to assess its potential as a biomarker for predicting stroke risk. Patients and Methods. The CHA2DS2-VASc score was used for thromboembolic risk stratification in AF according to American Heart Association (AHA) guidelines. We investigated the taxonomic and functional annotation of GM based on metagenomic data from 50 AF patients (32 with high thromboembolic risk (CHA2DS2-VASc score ≥2 (males) or CHA2DS2-VASc score ≥3 (females)) and 18 individuals with low thromboembolic risk (CHA2DS2-VASc score <2 (males) or CHA2DS2-VASc score <3 (females))). Results: The gut microbial diversity, composition, and function in AF were different in high and low CHA2DS2-VASc score groups. In high thromboembolic risk group, the abundance of Prevotella, Lachnospiraceae, and Eubacterium rectale, related to the production of short-chain fatty acids and anti-inflammatory were reduced (all P < 0.05). Furthermore, annotated by Kyoto Encyclopedia of Genes and Genomes (KEGG), a database of genes and genomes, the KEGG orthology-based scoring approach exhibited a significant association with thromboembolic risk in AF patients. Conclusions: Imbalance of GM and microbial dysfunction are involved in aggravated thromboembolic risk of AF.

Association of plasma Ninjurin-1 and SYNTAX score in patients with coronary artery disease.

BACKGROUND: Expression of nerve injury-induced protein 1 (Ninj1) is associated with several inflammatory disease. The soluble Ninj1 is an antiatherogenic protein. However, the role of plasma Ninj1 levels in patients with coronary artery disease (CAD) and its correlation with the severity of the disease remains unknown. This study investigated the association between plasma Ninj1 levels and the severity of coronary artery stenosis in patients with CAD. METHODS: A total of 207 subjects were recruited in this study. Blood samples were obtained to assess plasma Ninj1 level using enzyme-linked immunosorbent assay. The SYNTAX score calculated from baseline coronary angiography results was used to evaluate the severity of coronary artery stenosis. The least absolute shrinkage and selection operator (LASSO) regression analysis was performed to select the predictive factors. Then, a nomogram based on Ninj1 was constructed to predict the probability of CAD. RESULTS: Patients with CAD had significantly higher plasma Ninj1 than those without CAD (P < 0.001). A positive correlation was established between the Ninj1 levels and SYNTAX score (R = 0.352, P < 0.001). The multivariate logistical regression analysis indicated that plasma Ninj1 (P = 0.024) was an independent predictor of CAD occurrence after adjustment for clinical risk factors. The nomogram based on plasma Ninj1 level demonstrated good calibration and discrimination with the area under the curve 0.814. CONCLUSIONS: Plasma Ninj1 levels are increased in patients with CAD. Elevated levels of plasma Ninj1 are associated with CAD and the severity of coronary stenosis. A nomogram based on plasma Ninj1 and sectional clinical characteristics exerted a predictive potential for CAD.

A high-throughput analytical method for multiple DNA targets based on microdroplet PCR coupled with DGGE.

We developed a novel approach to analyze multiple DNA targets based on microdroplet PCR coupled with denaturing gradient gel electrophoresis (MPDG) to achieve high-throughput detection of biological samples. The target DNAs were preamplified using specific primers. Subsequently, the preamplified products were separated into individual microreactors for parallel amplification with high efficiency, avoiding the interference of different primers and templates, and preventing inconsistent amplification efficiency and non-specific amplification. The final products were analyzed using denaturing gradient gel electrophoresis (DGGE). Using genetically modified (GM) maize as samples, the MPDG method could be used for the simultaneous detection of three DNA targets with an absolute limit of detection of 0.5% (w/w), with no cross reaction with other non-GM samples. The simulated sample assay of MPDG suggested that the method is suitable for practical application. The MPDG approach, with high sensitivity and specificity, could play a crucial role in the field of nucleic acid detection.

Efficient fractionation of woody biomass hemicelluloses using cholinium amino acids-based deep eutectic solvents and their aqueous mixtures.

Novel and green cholinium amino acids-based deep eutectic solvents (DESs) and their aqueous mixtures were synthesized and employed in deconstructing poplar for hemicellulose fractionation. The effects of water content in DESs on hemicellulose dissociation and structural features were comprehensively investigated, along with the reusability of DESs for treatment. The integration of water into DESs could facilitate hemicellulose fractionation, and the cholinium lysine: urea with 5 wt% water (CL: U-5) demonstrated the best performance with a hemicellulose yield of 59.2%. Further structure analysis revealed that hemicelluloses with various branching degrees and molecular weights were obtained with varying water content of DESs. Furthermore, the CL: U-5 had recyclability and reusability with a 40.5% hemicellulose yield obtained after reused three times. The novel and eco-friendly cholinium amino acids-based DESs treatment provides an effective and sustainable strategy for hemicellulose fractionation from woody biomass.

Novel, recyclable Brønsted acidic deep eutectic solvent for mild fractionation of hemicelluloses.

Acidic deep eutectic solvents (DESs) are promising media for lignin valorization and cellulose conversion due to their good ability in efficient deconstruction of plant cell wall. However, hemicellulose extraction from lignocellulose using acidic DESs remains a challenge. Herein, novel and green Brønsted acidic DESs (BDESs) were synthesized from natural organic acids and common polyols and successively adopted to deconstruct corncob for mild fractionation of hemicelluloses. Oxalic acid (OA)-based BDESs were preferred for corncob processing due to the high solubility of xylan. The results revealed that the suitable acidity of DESs and mild temperature effectively avoided the over-degradation of hemicelluloses. The chemical composition and structural features of the recovered hemicelluloses were investigated systematically. Moreover, after ethylene glycol (EG)-OA BDES was recycled and reused three times, the extraction still resulted in a satisfactory hemicellulose yield. The novel and eco-friendly processing offers a practical and sustainable route for hemicellulose extraction in acidic condition.

Alkaline deep eutectic solvents as novel and effective pretreatment media for hemicellulose dissociation and enzymatic hydrolysis enhancement.

In recent years, deep eutectic solvents (DESs) are used for enhancing the enzymatic digestibility and lignin fractionation in pretreatment, while hemicellulosic fraction receives scant attention. Herein, we report a novel approach of applying alkaline deep eutectic solvents (ADESs) for dissociating hemicelluloses from woody biomass. Among these ADESs, choline chloride-monoethanolamine (C-M) was the most efficacious medium for deconstructing the recalcitrant structure of poplar and 63.3% of hemicelluloses was obtained at 80 °C. Structure analysis showed that the ADESs-extracted hemicelluloses retained partial of O-acetyl groups. Different ADESs could be used to obtain hemicelluloses with various degrees of branching. Furthermore, the enzymatic digestibility of cellulose was significantly increased by 6.6 times compared to that of the untreated poplar under the optimum conditions (C-M, 140 °C). This work provides a view on the dissociation behavior of hemicelluloses during ADESs pretreatment, which would be beneficial for devising DESs toward effective fractionation and comprehensive utilization of biomass.

Characterization of the complete chloroplast genome of Clerodendrum bungei Steud. (Lamiaceae).

Clerodendrum bungei Steud. is one kind of traditional medical herb which can be used for airway hyperreactivity treatment. In this study, the complete chloroplast genome sequence of C. bungei was assembled. Its complete circular chloroplast DNA length was 151,680 bp. The genome was made up of a large single-copy region of 83,189 bp, a small single-copy region of 17,311 bp, and a pair of inverted repeat regions of 25,590 bp. The genome totally encoded 130 genes, containing 85 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The phylogenetic analysis indicates that C. bungei belongs to the Lamiaceae family.

The complete chloroplast genome of Verbena officinalis, an herbal species of Verbenaceae family.

Verbena officinalis is one kind of traditional medical herb which has potential for multiple diseases' treatment. In this study, the complete chloroplast genome sequence of V. officinalis was assembled. Its complete circular chloroplast DNA length was 153,491 bp. The genome was made up of a large single-copy region of 84,518 bp, a small single-copy region of 17,357 bp, and a pair of inverted repeat regions of 25,808 bp. The genome totally encoded 128 genes, containing 83 protein-coding genes, 37 tRNA genes, and eight rRNA genes. Phylogenetic analysis indicates that V. officinalis belongs to the verbenaceae family.

Specific Decrease in B-Cell-Derived Extracellular Vesicles Enhances Post-Chemotherapeutic CD8+ T Cell Responses.

Systemic immunosuppression greatly affects the chemotherapeutic antitumor effect. Here, we showed that CD19+ extracellular vesicles (EVs) from B cells through CD39 and CD73 vesicle-incorporated proteins hydrolyzed ATP from chemotherapy-treated tumor cells into adenosine, thus impairing CD8+ T cell responses. Serum CD19+ EVs were increased in tumor-bearing mice and patients. Patients with fewer serum CD19+ EVs had a better prognosis after chemotherapy. Upregulated hypoxia-inducible factor-1α (HIF-1α) promoted B cells to release CD19+ EVs by inducing Rab27a mRNA transcription. Rab27a or HIF-1α deficiency in B cells inhibited CD19+ EV production and improved the chemotherapeutic antitumor effect. Silencing of Rab27a in B cells by inactivated Epstein-Barr viruses carrying Rab27a siRNA greatly improved chemotherapeutic efficacy in humanized immunocompromised NOD PrkdcscidIl2rg-/- mice. Thus, decreasing CD19+ EVs holds high potential to improve the chemotherapeutic antitumor effect.

A novel emulsion PCR method coupled with fluorescence spectrophotometry for simultaneous qualitative, quantitative and high-throughput detection of multiple DNA targets.

We constructed and validated a novel emulsion PCR method combined with fluorescence spectrophotometry (EPFS) for simultaneous qualitative, quantitative and high-throughput detection of multiple DNA targets. In a single reaction set, each pair of primers was labeled with a specific fluorophore. Through emulsion PCR, a target DNA was amplified in droplets that functioned as micro-reactors. After product purification, different fluorescent-labeled DNA products were qualitatively analyzed by the fluorescent intensity determination. The sensitivity and specificity of the system was examined using four kinds of genetically modified (GM) maize. The qualitative results revealed high specificity and sensitivity of 0.5% (w/w). In addition, the quantitative results revealed that the absolute limit of detection was 103 copies, showing good repeatability. Moreover, the reproducibility assays were further performed using four foodborne pathogenic bacteria to further evaluate the applicability of the system. Consequently, the same qualitative, quantitative and high-throughput results were confirmed with the four GM maize. To sum up, the new EPFS system is the first analytical technology of this kind that enables simultaneous qualitative, quantitative and high-throughput analysis of multiple genes.