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Arabidopsis thaliana MYB5 collaborates with TRANSPARENT TESTA GLABRA1 (TTG1) and basic-Helix-Loop-Helix (bHLH) transcription factors to regulate seed coat, trichome and root cell differentiation. Using a yeast two-hybrid system we show that the N-terminal region of MYB5 binds directly to the serine/threonine CASEIN KINASE2 BETA3 (CK2β3) subunit. Functions of the CASEIN KINASE2 (CK2) complex include facilitating phosphorylation of MYB transcription factors and cell cycle checkpoint regulatory proteins. Purified recombinant MYB5 protein was found to bind only weakly in vitro to the promoter of ALPHA/BETA ESTERASE/HYDROLASE4 (ABE4) , a known MYB5 target gene. We propose that phosphorylation of MYB5 facilitated by the MYB5-CK2β3 interaction enhances MYB5 binding to its target gene promoters., (Copyright: © 2021 by the authors.)

Epidemiological data suggest that the incidence of rheumatoid arthritis (RA) increases in postmenopausal women, which may be related to estrogen deficiency. Tissue acidosis is a common symptom of RA. Acid-sensitive ion channel 1a (ASIC1a), a member of the extracellular H + -activated cation channel family, could be activated by changes in extracellular pH and plays a crucial role in the pathogenesis of RA. As the only cellular component in cartilage tissue, chondrocytes play an extremely important role in maintaining cartilage tissue homeostasis. The aim of this study was to investigate whether estrogen could protect acid-stimulated chondrocytes by regulating the expression of ASIC1a and explore the possible mechanism. The results showed that estrogen could protect against acid-induced chondrocyte injury by reducing ASIC1a protein expression. Moreover, lysosome inhibitor chloroquine (CQ) and autophagy inhibitor 3-methyladeniine (3-MA) could reverse the reduction of ASIC1a protein caused by estrogen, indicating that autophagy-lysosome pathway contributes to estrogen-induced degradation of ASIC1a protein. Furthermore, the down-regulation of ASIC1a expression by estrogen was attenuated by MPP, a specific inhibitor of estrogen-related receptor-alpha (Esrra), indicating that Esrra is involved in the process of estrogen regulating the expression of ASIC1a. Additionally, adenosine 5'-monophosphate (AMP)-activated protein kinase/unc-51-like kinase 1 (AMPK-ULK1) signaling pathway was activated by estrogen treatment, which was abrogated by Esrra-silencing, and AMPK-specific inhibitor Compound C pretreatment could reduce estrogen-induced downregulation of ASIC1a protein. Taken together, these results indicate that estrogen could promote autophagy-lysosome pathway-dependent ASIC1a protein degradation and protect against acidosis-induced cytotoxicity, the mechanisms of which might relate to Esrra-AMPK-ULK1 signaling pathway., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Carex agglomerata C. B. Clarke is a sedge with excellent ornamental characters, it is an important ecosystem stabilizer. Here we report the complete chloroplast genome of C. agglomerata to provide a foundation for further phylogenetic studies on the Cyperaceae. The chloroplast (cp) genome is 184,157 bp in size and consists of a large single-copy (LSC) region 106,654 bp in length, a small single-copy (SSC) region of 36,099 bp, two inverted repeats (IR) regions each 20,702 bp. The total GC content of the cp genome is 33.9% with the LSC, SSC, and IR regions 32, 32.5, and 42.9%, respectively. The cp genome contains 128 genes, including 80 protein-coding, 40 tRNA, and eight rRNA genes. The phylogenetic analysis showed C. agglomerata is in a clade with Carex neurocarpa Maxim and Carex siderosticta Hance. This study provides a basis for further phylogenetic studies of Carex., Competing Interests: The authors report no potential conflict of interest., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Chemical control of fungal spoilage of postharvest cereal grains is an important strategy for the management of grain storage. Here, the potential antifungal activity of 1-nonanol, a main component of cereal volatiles, against Aspergillus flavus was studied. The growth of A. flavus was completely inhibited by 0.11 and 0.20 μL/mL 1-nonanol at vapor and liquid contact phases, respectively. Metabolomic analysis identified 135 metabolites whose expression was significantly different between 1-nonanol-treated and untreated A. flavus. These metabolites were involved in the tricarboxylic acid cycle, amino acid biosynthesis, protein degradation and absorption, aminoacyl-tRNA biosynthesis, mineral absorption, and in interactions with ABC transporters. Biochemical validation confirmed the disruptive effect of 1-nonanol on A. flavus growth, as indicated by the leakage of intracellular electrolytes, decreased succinate dehydrogenase, mitochondrial dehydrogenase, and ATPase activity, and the accumulation of reactive oxygen species. We speculated that 1-nonanol could disrupt cell membrane integrity and mitochondrial function and might induce apoptosis of A. flavus mycelia. Simulated grain storage experiments showed that 1-nonanol vapor, at a concentration of 264 μL/L, completely inhibited A. flavus growth in wheat, corn, and paddy grain with an 18% moisture content. This study provides new insights into the antifungal mechanism of 1-nonanol against A. flavus, indicating that it has a promising potential as a bio-preservative to prevent fungal spoilage of postharvest grains. KEY POINTS: • 1-Nonanol showed higher antifungal activity against A. flavus. • The antifungal mechanisms of 1-nonanol against A. flavus were revealed. • 1-Nonanol could damage cell membrane integrity and mitochondrial function., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Aspergillus flavus is a notorious saprophytic fungus that compromises the quantity and quality of postharvest grains and produces carcinogenic aflatoxins. The natural compound hexanal disrupts cell membrane synthesis and mitochondrial function and induces apoptosis in A. flavus; here, we investigated the molecular mechanisms underlying these effects. The minimum inhibition and fungicidal concentration (MIC and MFC) of hexanal against A. flavus spores were 3.2 and 9.6 μL/mL, respectively. Hexanal exposure resulted in abnormal spore morphology and early spore apoptosis. These changes were accompanied by increased reactive oxygen species production, reduced mitochondrial membrane potential, and DNA fragmentation. Transcriptomic analysis revealed that hexanal treatment greatly altered the metabolism of A. flavus spores, including membrane permeability, mitochondrial function, energy metabolism, DNA replication, oxidative stress, and autophagy. This study provides novel insights into the mechanism underlying the antifungal activity of hexanal, suggesting that hexanal can be used an anti-A. flavus agent for agricultural applications. KEY POINTS: • Hexanal exposure resulted in abnormal spore morphology. • The apoptotic characteristics of A. flavus were induced after hexanal treatment. • Hexanal could change the expression of key A. flavus growth-related genes., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

DNA methylation is a crucial regulatory mechanism in many biological processes. However, limited studies have dissected the contribution of DNA methylation to sexual differentiation in dioecious plants. In this study, we investigated the variances in methylation and transcriptional patterns of male and female flowers of garden asparagus. Compared with male flowers, female flowers at the same stages showed higher levels of DNA methylation. Both male and female flowers gained DNA methylation globally from the premeiotic to meiotic stages. Detailed analysis revealed that the increased DNA methylation was largely due to increased CHH methylation. Correlation analysis of differentially expressed genes and differentially methylated regions suggested that DNA methylation might not have contributed to the expression variation of the sex-determining genes SOFF and TDF1 but probably played important roles in sexual differentiation and flower development of garden asparagus. The upregulated genes AoMS1, AoLAP3, AoAMS, and AoLAP5 with varied methylated CHH regions might have been involved in sexual differentiation and flower development of garden asparagus. Plant hormone signaling genes and transcription factor genes also participated in sexual differentiation and flower development with potential epigenetic regulation. In addition, the CG and CHG methylation levels in the Y chromosome were notably higher than those in the X chromosome, implying that DNA methylation might have been involved in Y chromosome evolution. These data provide insights into the epigenetic modification of sexual differentiation and flower development and improve our understanding of sex chromosome evolution in garden asparagus., (© 2021. The Author(s).)

Background: Aldo-keto reductases-catalyzed transformations of ketones to chiral alcohols have become an established biocatalytic process step in the pharmaceutical industry. Previously, we have discovered an aldo-keto reductase (AKR) from Kluyveromyces marxianus that is active to the aliphatic tert-butyl 6-substituted (5R/S)-hydroxy-3-oxohexanoates, but it is inactive to aromatic ketones. In order to acquire an excellent KmAKRmutant for ensuring the simultaneous improvement of activity-thermostability toward tert-butyl 6-cyano-(5R)-hydroxy-3-oxohexanoate ((5R)-1) and broadening the universal application prospects toward more substrates covering both aliphatic and aromatic ketones, a fluorescence-based high-throughput (HT) screening technique was established., Main Methods and Major Results: The directed evolution of KmAKR variant M5 (KmAKR-W297H/Y296W/K29H/Y28A/T63M) produced the "best" variant M5-Q213A/T23V. It exhibited enhanced activity-thermostability toward (5R)-1, improved activity toward all 18 test substrates and strict R-stereoselectivity toward 10 substrates in comparison to M5. The enhancement of enzymatic activity and the extension of substrate scope covering aromatic ketones are proposed to be largely attributed to pushing the binding pocket of M5-Q213A/T23V to the enzyme surface, decreasing the steric hindrance at the entrance and enhancing the flexibility of loops surrounding the active center. In addition, combined with 0.94 g dry cell weight (DCW) L -1 glucose dehydrogenase from Exiguobacterium sibiricum (EsGDH) for NADPH regeneration, 2.81 g DCW L -1 M5-Q213A/T23V completely converted (5R)-1 of up to 450 g L -1 at 120 g g -1 substrates/catalysts (S/C), yielding the corresponding optically pure tert-butyl 6-cyano-(3R,5R)-dihydroxyhexanoate ((3R,5R)-2, > 99.5% d.e. p ) with a space-time yield (STY) of 1.08 kg L -1 day -1 ., Conclusions: A fluorescence-based HT screening system was developed to tailor KmAKR's activity, thermostability and substrate scope. The "best" variant M5-Q213A/T23V holds great potential application for the synthesis of aliphatic/aromatic R-configuration alcohols., (© 2021 Wiley-VCH GmbH.)

Background: Intestinal barrier breakdown, a frequent complication of intestinal ischemia-reperfusion (I/R) including dysfunction and the structure changes of the intestine, is characterized by a loss of tight junction and enhanced permeability of the intestinal barrier and increased mortality. To develop effective and novel therapeutics is important for the improvement of outcome of patients with intestinal barrier deterioration. Recombinant human angiopoietin-like protein 4 (rhANGPTL4) is reported to protect the blood-brain barrier when administered exogenously, and endogenous ANGPTL4 deficiency deteriorates radiation-induced intestinal injury., Aim: To identify whether rhANGPTL4 may protect intestinal barrier breakdown induced by I/R., Methods: Intestinal I/R injury was elicited through clamping the superior mesenteric artery for 60 min followed by 240 min reperfusion. Intestinal epithelial (Caco-2) cells and human umbilical vein endothelial cells were challenged by hypoxia/ reoxygenation to mimic I/R in vitro ., Results: Indicators including fluorescein isothiocyanate-conjugated dextran (4 kilodaltons; FD-4) clearance, ratio of phosphorylated myosin light chain/total myosin light chain, myosin light chain kinase and loss of zonula occludens-1, claudin-2 and VE-cadherin were significantly increased after intestinal I/R or cell hypoxia/reoxygenation. rhANGPTL4 treatment significantly reversed these indicators, which were associated with inhibiting the inflammatory and oxidative cascade, excessive activation of cellular autophagy and apoptosis and improvement of survival rate. Similar results were observed in vitro when cells were challenged by hypoxia/reoxygenation, whereas rhANGPTL4 reversed the indicators close to normal level in Caco-2 cells and human umbilical vein endothelial cells significantly., Conclusion: rhANGPTL4 can function as a protective agent against intestinal injury induced by intestinal I/R and improve survival via maintenance of intestinal barrier structure and functions., Competing Interests: Conflict-of-interest statement: The authors declare that they have no competing interests., (©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.)

Cellular senescence has been considered an important driver of many chronic lung diseases. However, the specific mechanism of cellular senescence in silicosis is still unknown. In the present study, silicotic rats and osteoclast stimulatory transmembrane protein ( Ocstamp ) overexpression of MLE-12 cells were used to explore the mechanism of OC-STAMP in cellular senescence in alveolar epithelial cell type II (AEC2). We found an increasing level of OC-STAMP in AEC2 of silicotic rats. Overexpression of Ocstamp in MLE-12 cells promoted epithelial-mesenchymal transition (EMT), endoplasmic reticulum (ER) stress, and cellular senescence. Myosin heavy chain 9 (MYH9) was a potential interacting protein of OC-STAMP. Knockdown of Ocstamp or Myh9 inhibited cellular senescence in MLE-12 cells transfected with pcmv6- Ocstamp . Treatment with 4-phenylbutyrate (4-PBA) to inhibit ER stress also attenuated cellular senescence in vitro or in vivo . In conclusion, OC-STAMP promotes cellular senescence in AEC2 in silicosis., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2021 Tian Li et al.)

Purpose: To analyze the characteristics, related risk factors, and prognosis of suprachoroidal hemorrhage (SCH) associated with pars plana vitrectomy (PPV)., Methods: Cases of SCH associated with PPV excluding trauma were retrospectively analyzed in Beijing Tongren Hospital between January 2010 and June 2020. The data collected included general data, myopia status, axial length, state of the crystalline lens, SCH onset time, range, treatment method, visual prognosis, and methods of operation and anesthesia. Patients were divided into those with SCH related to the first PPV (Group 1), and SCH related to second intraocular surgery in the vitrectomized eye (Group 2). Patients were also classified by the SCH onset time into either the expulsive suprachoroidal hemorrhage group (ESCH) and the delayed suprachoroidal hemorrhage group (DSCH). The general data, related risk factors, and the visual prognosis of SCH in the different groups were analyzed., Results: SCH associated with PPV was studied in 28 cases with an incidence of 0.06 %; 16 males and 12 females. The mean age of the patients was (53.51 ± 10.21) years old, the mean follow-up time was (24.94 ± 14.60) days, and the mean axial length was (28.21 ± 3.14) mm. Of these cases, 21 were classified as high myopia, 25 as aphakia/ pseudophakic, and 7 as focal hemorrhage. Silicone oil removal occurred in 12 cases (43 %). Patients in Group 2 were younger than Group 1 (P = 0.005). In terms of treatment and prognosis, 5 eyes were simply closely observed, 4 were given single suprachoroidal drainage, 15 were given suprachoroidal drainage combined with silicone tamponade, 2 underwent anterior chamber puncture, and 2 gave up treatment. A follow-up vision: NLP ~ 20/30; among them, 2 eyes with NLP (7.14 %), 6 of ≥ 20/200 (21.43 %). The final outcomes presented a significantly positive correlation with baseline vision but no significant correlation with age or axial length., Conclusions: SCH has a higher incidence rate after a second intraocular surgery in a vitrectomized eye which is associated with the lack of vitreous support and easier fluctuation of intraocular pressure. SCH associated with PPV is more localized and has a relatively good prognosis; high myopia and aphakic/ pseudophakic eyes are risk factors. Active treatment can effectively improve visual prognosis., Trial Registration: Retrospective case series study, not applicable., (© 2021. The Author(s).)

Objectives: To explore factors affecting the efficacy of Bernese periacetabular osteotomy for the treatment of hip dysplasia., Methods: A retrospective study was conducted on 44 patients with hip dysplasia who underwent Bernese periacetabular osteotomy with a modified Smith-Peterson approach between January 2017 and November 2019. Among them, 40 were women and four were men. The average age was 31.2 ± 9.4. Preoperative and postoperative imaging parameters were measured. The acetabular top tilt angle, lateral central edge angle, acetabular abduction angle, femoral head extrusion index, sphericity index of femoral head, Shenton line, Tonnis grade of osteoarthritis, joint congruency, p/a ratio, acetabular anteversion angle, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scale scores, and modified Harris hip score (MHHS) were observed. MHHS were divided into three clinically relevant categories: poor (<70 points), good (70-85 points), and excellent (86-91 points). Patient demographic data, as well as preoperative and postoperative radiographic parameters, were subjected to univariate logistic regression analysis. Multiple regression analysis was used to determine factors influencing postoperative MHHS., Results: The follow-up time was 1.0-3.9 years after surgery, with an average of 1.6 years. By the last follow-up, MHHS increased from 70 points before surgery to 91 points after surgery (P < 0.001), WOMAC pain score decreased from 4 points before surgery to 0 points after surgery (P < 0.001). WOMAC functional score decreased (Preoperative: 18.0 [4.0]; Postoperative: 4.0 [0], P = 0.004). Six patients had sensory disturbance of the lateral femoral cutaneous nerve, four of which recovered completely during follow-up. No other complications related to surgical approach, osteotomy, acetabular displacement, acetabular fixation, and postoperative stage were found. There was no significant vascular, nerve, or visceral injuries in any of the patients. On multiple regression analysis, the probability of the postoperative modified Harris hip score of a hip joint with a preoperative lateral center edge angle ≥4.5° being classified as excellent was six times that of angles <4.5° (Exp[β]: 6.249, 95% CI: 1.03-37.85, P = 0.046). Regression analysis of other factors found no significant correlation with postoperative functional scores., Conclusion: Overall functional scores post-PAO significantly improved, and pain symptoms were significantly reduced. Patients with a preoperative lateral center edge angle ≥4.5° had better joint function after surgery., (© 2021 The Authors. Orthopaedic Surgery published by Chinese Orthopaedic Association and John Wiley & Sons Australia, Ltd.)

Microbial production of industrial chemicals is a sustainable approach to reduce the dependence on petroleum-based chemicals such as acids, alcohols, and amines, in which the cadaverine is a natural diamide and serves as one of the key monomers for biopolymer production. In this study, the constitutive promoter J23100 driven lysine decarboxylase (CadA) for cadaverine production was established and compared in different Escherichia coli strains. The best chassis designed as JW, expressed the highest amount of CadA by using J23100 promoter, showing stable and high copy numbers (i.e., PCN > 100) when culture in the antibiotic-free medium. JW attained a CadA activity of 167 g-DAP/g-DCW-h and had the maximum biocatalyst of 45.6 g-DCW/L in fed-batch fermentation. In addition, JW was able to convert 2.5 M L-lysine to 221 g/L cadaverine, with 86 % yield and 55.3 g/L-h productivity. The whole-cell biocatalyst could be reused over four times at an average of 97 % conversion when supplied half of fresh cells in the reaction. This work developed a stable, constitutive expression, long-term preservation, high-level expression of CadA for DAP production, and paved an alternative opportunity of bio-nylon for industry in the future., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Dendrobium viroid (DVd) was first reported in China in 2020, and it is the only viroid known to infect Orchidaceae family plants. In this study, we developed a simple reverse transcription-polymerase chain reaction (RT-PCR) method for the rapid detection of DVd in Dendrobium plants. When extracting the sap template from the leaves, they are first clamped between two layers of plastic film, and the sap is pressed out and collected with a pipette. Using this sap, DVd was detected by dot-blot and RT-PCR methods and, the expected amplicons were confirmed by sequencing analysis. The batch analysis of field samples revealed that this method can be used to detect DVd rapidly. The detection method also reduces cross-contamination between different samples and minimizes false positives. Thus, this sap-direct RT-PCR method allows effective and rapid DVd detection in the study of Orchidaceae plants., (© 2021 The Society for Applied Microbiology.)

Introduction: The complex electrophysiological phenomena related to the atrioventricular node (AVN) are due to its complex anatomical structures. Aside from the inferior nodal extension (INE), other node-like tissues, such as the retroaortic node (RN), have been described less extensively and may also share the mechanism of normal conduction and abnormal conduction in AVN re-entrant tachycardia., Methods: High-density sections of the entire AVN were obtained from rats and rabbits. Fibrosis was analyzed by Masson's trichrome staining. Connexin (Cx43, Cx40, and Cx45) and ion channel (Na v 1.5, Ca v 3.1, and HCN4) proteins were immunohistochemically labeled for the analysis of tissue features. Three-dimensional (3D) reconstruction of the AV junction was performed to clarify the relationships among different structures., Results: The RN expressed the same connexin isoforms as the compact node (CN) and INE. Na v 1.5 labeling was observed at low levels in the CN, RN, and INE, where Ca v 3.1 and HCN4 were expressed. The CN connected with the RN in a narrow strip pattern at the start of the CN. The RN presented as a shuttle shape and was the only tissue directly connected with the atrium in the anterior septum., Conclusion: The RN connects with the AVN anatomically, suggesting that direct electrical conduction occurs between them. The entrance of the atria into the AVN is distal to the RN, which may form the fast AVN pathway., (© 2021 Wiley Periodicals LLC.)

Objective: To explore the potential mechanism of Huanglian Jiedu Decoction (HJD) treatment and prevention of metastatic Cutaneous Melanoma (CM) occurrence and metastasis based on network pharmacological methods and immune infiltration analysis., Methods: The GEO database was used to obtain metastatic CM disease targets, the TCMSP database and the HERB database were used to obtain HJD action targets, core genes were screened by protein interaction network, and the potential mechanism of HJD in the treatment of metastatic CM was explored by enrichment analysis, prognostic analysis and immune infiltration analysis., Results: HJD treatment of metastatic CM involved 60 targets, enrichment analysis showed that HJD treatment of metastatic CM involved Chemokine signaling pathway, NF-kappa B signaling pathway, and Fluid shear stress and atherosclerosis, etc. Prognostic analysis revealed that HJD had a certain ability to improve the prognosis of metastatic CM patients. Immune infiltration analysis showed that HJD could inhibit the immune cell infiltration of metastatic CM patients by acting on related targets., Conclusions: Our study identified the potential mechanism of HJD in the treatment of metastatic CM through network pharmacology, and revealed the mechanism of HJD in the prevention of Skin Cutaneous Melanoma metastasis through immune infiltration analysis and prognostic analysis., Competing Interests: The authors declare that they have no conflicts of interests., (Copyright © 2021 Ding Li et al.)

Main Conclusion: Transcriptomic analyses identified anther-expressed genes in wheat likely to contribute to heat tolerance and hence provide useful genetic markers. The genes included those involved in hormone biosynthesis, signal transduction, the heat shock response and anther development. Pollen development is particularly sensitive to high temperature heat stress. In wheat, heat-tolerant and heat-sensitive cultivars have been identified, although the underlying genetic causes for these differences are largely unknown. The effects of heat stress on the developing anthers of two heat-tolerant and two heat-sensitive wheat cultivars were examined in this study. Heat stress (35 °C) was found to disrupt pollen development in the two heat-sensitive wheat cultivars but had no visible effect on pollen or anther development in the two heat-tolerant cultivars. The sensitive anthers exhibited a range of developmental abnormalities including an increase in unfilled and clumped pollen grains, abnormal pollen walls and a decrease in pollen viability. This subsequently led to a greater reduction in grain yield in the sensitive cultivars following heat stress. Transcriptomic analyses of heat-stressed developing wheat anthers of the four cultivars identified a number of key genes which may contribute to heat stress tolerance during pollen development. Orthologs of some of these genes in Arabidopsis and rice are involved in regulation of the heat stress response and the synthesis of auxin, ethylene and gibberellin. These genes constitute candidate molecular markers for the breeding of heat-tolerant wheat lines.

Hexanal, a natural volatile organic compound, exerts antifungal activity against Aspergillus flavus; however, the mechanisms underlying these effects are unclear. In this study, we found that the growth of A. flavus mycelium was completely inhibited following exposure to 0.4 μL/mL hexanal (minimal inhibitory concentration). A detailed metabolomics survey was performed to identify changes in metabolite production by A. flavus cells after exposure to 1/2 the minimal inhibitory concentration of hexanal for 6 h, which revealed significant differences in 70 metabolites, including 20 upregulated and 50 downregulated metabolites. Among them, levels of L-malic acid, α-linolenic acid, phosphatidylcholine, D-ribose, riboflavin, D-mannitol, D-sorbitol, and deoxyinosine were significantly reduced. The metabolomics results suggest that the metabolites are mainly involved in the tricarboxylic acid cycle (TCA), ABC transport system, and membrane synthesis in A. flavus cells. Hexanal treatment reduced succinate dehydrogenase and mitochondrial dehydrogenase activity and stimulated superoxide anion and hydrogen peroxide accumulation in A. flavus mycelia. Increases in the electric conductivity and A 260nm of the culture supernatant indicated cell membrane leakage. Therefore, hexanal appears to disrupt cell membrane synthesis, induce mitochondrial dysfunction, and increase oxidative stress in A. flavus mycelia. KEY POINTS: • Metabolite changes of A. flavus mycelia were identified after hexanal treatment. • Most differential metabolites were downregulated in hexanal-treated A. flavus. • An antifungal model of hexanal against A. flavus was proposed.

The discovery of efficient and specific HIV-latency-reversing agents is critical for HIV therapy. Here, we developed wikstroelide E, a daphnane diterpene from the buds of Wikstroemia chamaedaphne , as a potential HIV-latency-reversing agent that is 2500-fold more potent than the drug prostratin. Based on transcriptome analysis, the underlying mechanism was that wikstroelide E regulated the MAPK, PI3K-Akt, JAK-Stat, TNF, and NF-κB signaling pathways. We clearly demonstrated that wikstroelide E reversed latent HIV infection by activating PKC-NF-κB signals, serving as a proxy for verifying the transcriptome data. Strikingly, the Tat protein contributes to the robust activation of latent HIV in wikstroelide-E-treated cells, producing an unexpected latency-reversing effect against latent HIV. This study provides the basis for the potential development of wikstroelide E as an effective HIV-latency-reversing agent.

Objective: This study aimed to use an air-liquid interface (ALI) exposure system to simulate the inhalation exposure of motorcycle exhaust particulates (MEPs) and then investigate the benchmark dose (BMD) of MEPs by evaluating cell relative viability (CRV) in lung epithelial BEAS-2B cells., Methods: The MEPs dose was characterized by measuring the number concentration (NC), surface area concentration (SAC), and mass concentration (MC). BEAS-2B cells were exposed to MEPs at different concentrations via ALI and CRV was determined using Cell Counting Kit (CCK-8) assay. BMD software was applied to calculate BMD and the lower limit of benchmark dose (BMDL) according to Akaike Information Coefficient (AIC), with P -value based on Hill, Linear, Polynomial, and Power model., Results: Our results reveal that BMD of NC and SAC were estimated by the best-fitting Hill model, while MC was estimated by Polynomial model. The BMDL for CRV following ALI exposure to MEPs were as follows: 364.2#/cm 3 for NC; 0.662 × 10 7 nm 2 /cm 3 for SAC; and 0.278 μg/m 3 for MC., Conclusion: These results indicate that MEPs exposure via ALI system induces a dose-dependent decrease of CRV and provides the potential exposure threshold of MEPs in a lung cell model., (Copyright © 2021 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.)

Objectives: Our aim was to investigate the prevalence and predictive variables of sarcopenia., Methods: We recruited participants from the Peking Union Medical College Hospital Multicenter Prospective Longitudinal Sarcopenia Study (PPLSS). Muscle mass was quantified using bioimpedance, and muscle function was quantified using grip strength and gait speed. Logistic regression revealed the relationships between sarcopenia and nutritional, lifestyle, disease, psychosocial and physical variables., Results: The prevalence of sarcopenia and sarcopenic obesity was 9.2%-16.2% and 0.26%-9.1%, respectively. Old age, single status, undernourishment, higher income, smoking, low physical activity, poor appetite and low protein diets were significantly associated with sarcopenia. Multiple logistic regression analysis showed that age was a risk factor for all stages of sarcopenia, and participants above 80 years were greater than fivefold more susceptible to sarcopenia, while lower physical activity was an independent risk factor. The optimal cut-off value for age was 71 years, which departs from the commonly accepted cut-off of 60 years. Female participants were greater than twofold less susceptible to sarcopenia than male participants. The sterol derivative 25-hydroxyvitamin D was associated with fourfold lower odds of sarcopenia in male participants. Several protein intake variables were also correlated with sarcopenia. Based on these parameters, we defined a highly predictive index for sarcopenia., Conclusions: Our findings support a predictive index of sarcopenia, which agglomerates the complex influences that sterol metabolism and nutrition exert on male vs female participants., (© 2021 The Authors. Cell Proliferation Published by John Wiley & Sons Ltd.)

t-Butyl 6-cyano-(3R,5R)-dihydroxyhexanoate ((3R,5R)-2) is an advanced chiral diol intermediate of the cholesterol-lowering drug atorvastatin. KmAKR M5 (W297H/Y296W/K29H/Y28A/T63M) constructed in our previous work, displayed good biocatalytic performance on (3R,5R)-2. In the present work, stepwise evolution was applied to further enhance the thermostability and activity of KmAKR M5 . For thermostability enhancement, N109 and S196 located far from the active site were picked out by structure-guided consensus engineering, and mutated by site-directed mutagenesis (SDM). For catalytic efficiency improvement, the residues A30 and T302 adjacent to the substrate-binding pocket were subjected to site-saturation mutagenesis (SSM). As a result, the "best" mutant KmAKR M9 (W297H/Y296W/K29H/Y28A/T63M/A30P/T302S/N109K/S196C) was developed, of which T 50 15 and T m were 5.0 °C and 8.2 °C higher than those of KmAKR M5 . Moreover, compared to KmAKR M5 , KmAKR M9 displayed a 1.9-fold (846 vs 2436 min) and 6.7-fold (126 vs 972 min) longer half-lives at 40 and 50 °C, respectively. Structural analysis suggested that beneficial mutations introduced additional hydrophobic interactions and hydrogen bonds, contributing rigidification of the flexible loops and the increase of internal forces, hence increasing the thermostability and activity. 5 g DCW (dry cell weight) L -1 KmAKR M9 completely reduced 350 g L -1 t-butyl 6-cyano-(5R)-hydroxy-3-oxo-hexanoate ((5R)-1), within 3.7 h at 40 °C, yielding optically pure (3R,5R)-2 (d.e. p  > 99.5%) with a space-time yield (STY) of 1.82 kg L -1 d -1 . Hence, KmAKR M9 is a robust biocatalyst for the synthesis of (3R,5R)-2., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Awareness is one of the first steps in a lifestyle modification process. Since older adults are at a higher risk of hypertension, maintaining low sodium intake is of utmost importance. Yet, it is uncertain if older adults have an awareness of their sodium intake. This study aimed to explore self-perceived sodium intake among older Chinese people and compare it with various measures of sodium intake. A cross-sectional study was conducted in Hong Kong in 2015. Data were collected from 24-h urinary sodium excretion (UNa24) measurements, dietary sodium intake by 24-h diet recalls and self-perceived salt intake levels among the older Hong Kong population. Kappa statistics and a paired t -test were used to compare the different measures. Fifty-nine participants provided complete 24-h urine samples. The mean (±SD) UNa24 of participants was 2846 ± 1253 mg, of which 74.6% exceeded the recommended daily sodium intake. About 87.8% of participants with UNa24 values >2000 mg perceived their sodium intake as too little or just right. The kappa statistic between UNa24 and self-perception was insignificantly different from zero (κ = -0.003, p -value = 0.909). The UNa24 estimates were significantly higher than those estimated from the 24-h diet recalls by 1203 mg ( p -value <0.001). The findings imply that the low awareness of excessive sodium intake should be raised among older people to promote a healthy intake of sodium.

Two unprecedented and stable metal-organic frameworks, {[Co 2 (H 2 O) 2 (L)(OH)]·2.5H 2 O·0.5DMF} n ( 1 ) and {[Ni 2 (H 2 O) 2 (L)(OH)]·1.75H 2 O} n ( 2 ), have been synthesized (H 3 L = 5-(5-carboxy-pyridin-3-yloxy)-isophthalic acid, DMF = N , N -dimethylformamide). Structural analysis shows that 1 and 2 are heteronuclear isomorphous, possessing a three-dimensional (3D) (4,8)-connected flu/fluorite topological framework formed through the interconnection of tetranuclear butterfly {M 4 (COO) 6 (OH) 2 } clusters and the ligands. Although the frameworks of these two compounds are similar, their magnetic properties are different. Compound 1 exhibits an antiferromagnetic interaction in the high-temperature region, while 2 shows a weak ferromagnetic interaction in the whole-temperature region. Furthermore, considering the presence of hydroxyl groups and water molecules in the frameworks, we tested their proton conductivity. The efficient proton transfer pathway in the framework endowed 1 and 2 with excellent proton conductivities of 9.07 × 10 -5 and 1.29 × 10 -4 S·cm -1 at 363 K and 98% relative humidity (RH), respectively., Competing Interests: The authors declare no competing financial interest., (© 2021 The Authors. Published by American Chemical Society.)

This study investigated the risk of insomnia and hypnotics use among emergency physicians. This cross-sectional study recruited physicians working in Taiwanese hospitals in 2015 and the general population as the participants. Data from 1,097 emergency physicians obtained from the National Health Insurance Research Database were grouped into the case group, whereas 14,112 nonemergency physicians and 4,388 people from the general population were categorized into the control groups. This study used logistic regression and conditional logistic regression to compare the risks of insomnia between emergency and nonemergency physicians and between emergency physicians and the general population, respectively. The prevalence of insomnia among emergency physicians, nonemergency physicians and general population was 5.56%, 4.08%, and 1.73%, respectively. Compared with nonemergency physicians and the general population, emergency physicians had a significantly higher risk of insomnia. The proportions of emergency physicians, nonemergency physicians, and general population using hypnotics were 19.96%, 18.24%, and 13.26%, respectively. Among emergency physicians who used hypnotics, 49.77%, 25.57%, and 24.66% used only benzodiazepines, only nonbenzodiazepines, and both benzodiazepines and nonbenzodiazepines, respectively. Nonpharmacological interventions to improve insomnia and reminder of safe use of hypnotics to emergency physicians can serve as references for hospitals in developing health-promoting activities.

An oral multi-unit delivery system was developed by incorporating the nanoparticle (NP) into the nanofiber mat and its efficiency for intestinal-specific delivery and controlled release of a peptide (insulin) was investigated. Initially, the influence of deacetylation degree (DD) of chitosan and ionic gelation methods on the properties of NPs was studied. High DD (95%) chitosan was attributed to higher encapsulation efficiency and stability when crosslinked with polyanion tripolyphosphate. Subsequently, the multi-unit system was fabricated using a pH-sensitive polymer (sodium alginate) as the coating layer to further encapsulate the NP. Fiber mat with an average diameter of 481 ± 47 nm could significantly decrease the burst release of insulin in acidic condition and release most amount of insulin (>60%) in the simulated intestinal medium. Furthermore, the encapsulated peptide remained in good integrity. This multi-unit carrier provides the better-designed vehicle for intestinal-specific delivery and controlled release of the peptide., (© 2021 IOP Publishing Ltd.)

To explore the potential targets underlying the effect of rosuvastatin on heart failure (HF) by utilizing a network pharmacology approach and experiments to identify the results. PharmMapper and other databases were mined for information relevant to the prediction of rosuvastatin targets and HF-related targets. Then, the rosuvastatin-HF target gene networks were created in Cytoscape software. Eventually, the targets and enriched pathways were examined by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Furthermore, we constructed an HF animal model and used rosuvastatin to treat them, identifying the changes in heart function and related protein expression. We further used different cells to explore the mechanisms of rosuvastatin. Thirty-five intersection targets indicated the therapeutic targets linked to HF. GO analysis showed that 481 biological processes, 4 cellular components and 23 molecular functions were identified. KEGG analysis showed 13 significant treatment pathways. In animal experiments, rosuvastatin significantly improved the cardiac function of post-myocardial infarction mice and prevented the development of HF after myocardial infarction by inhibiting IL-1Β expression. Cell experiments showed that rosuvastatin could reduce the expression of IL-1B in HUVEC and THP-1 cells. The therapeutic mechanism of rosuvastatin against HF may be closely related to the inhibition of the expression of apoptosis-related proteins, inflammatory factors, and fibrosis-related genes. However, IL-1Β is one of the most important target genes., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Soil bacterial diversity is a key factor for the maintanence of forest ecosystem function. Soil bacterial community would change along forest succession. We analyzed the variations of soil bacterial diversity and community composition at different successional stages in the Pinus yunnanensis forest, which would help understand the mechanism underlying forest restoration. We investigated soil bacterial diversity, community composition, and effect factors at different successional stages (including coniferous forest, mixed coniferous and broadleaf forest, and evergreen broadleaf forest) using Illumina Hiseq platform. The results showed that OTUs, Chao1 index, Ace index, and Shannon index of soil bacterial community decreased with the process of secondary succession. The highest soil bacterial total OTUs, richness, and complexity appeared at early successional stage. Soil bacterial community composition varied across different stages, with the mixed coniferous and broadleaf forest showing largest variation. Proteobacteria and Acidobacteria were common dominant phyla at secondary successional stages. Actinobacteria, Chloroflexi, and Patescibacteria were dominant phyla at the early successional stage, the abundance of which decreased with successional process in the P. yunnanensis forest. Proteobacteria and WPS-2 increased with the succession. Soil pH and tree species richness were key factors in driving soil bacterial community structure. The soil bacterial diversity decreased with forest succession, while the variations of soil bacterial community composition became larger.

A 44-year-old woman presented to the Emergency Department with abdominal pain. She had a history of fibroids and no prior surgeries. Ultrasonography and CT imaging revealed a small bowel obstruction and massive uterine fibroids. The patient required laparotomy to relieve the intestinal obstruction after conservative therapy failed. Massive uterine fibroids is a rare cause of small bowel obstruction which requires the vigilance of Emergency Medicine physicians., Competing Interests: Declaration of Competing Interest The authors do not have any declarations of interest to disclose pertaining to this manuscript., (Copyright © 2020 Elsevier Inc. All rights reserved.)

A subchronic toxicity study was conducted in Wistar rats to evaluate the potential health effects of genetically modified (GM) herbicide-tolerant soybean DAS-68416-4. Rats were fed with diets containing toasted meal produced from GM soybean engineered with aad-12 and pat genes or containing non-GM soybean at a dose of 30.0, 15.0, or 7.5%,w/w% and 0% (control group) for 90 consecutive days. Animals were evaluated for general behavior, body weight gain, food consumption, food use efficiency, etc. At the middle and end of the study, blood and serum samples were collected for routine and biochemical assays. Internal organs were taken for calculating relative weights and doing histopathological examination. The rats were active and healthy without any abnormal symptoms during the entire study period. No biological differences in hematological or biochemical indices were detected. No histopathological changes were observed. Under the conditions of this study, herbicide-tolerant soybean DAS-68416-4 did not cause any treatment-related effects in Wistar rats following 90 days of dietary administration., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The Coronavirus disease 2019 (COVID-19) outbreak has been brought under control through a nationwide effort, and now it has become a global pandemic and the situation seems grim. We summarized the measures taken in Wuhan and analyzed the effects to comprehensively describe the factors involved in controlling the COVID-19 in China. In China, several measures such as the lockdown of Wuhan, restriction of traffic and communities, increasing hospital beds, nationwide support from medical staff, epidemic prevention equipment and supplies, and establishment of makeshift shelter hospitals have been taken. The lockdown of Wuhan reduced the propagation of cases to other cities in Hubei province and throughout China, traffic and community restrictions reduced the flow of population and the spread of disease, increasing wards and beds and medical personnel reduced the incidence of severe cases and mortality, the establishment of the Fangcang shelter hospitals provided a good isolation and monitoring environment, and further reduced the spread and fatality of the disease. The fact that China was able to control the spread of COVID-19 within three months without a specific drug or vaccine suggests that these measures are more adequate and effective.

Members of the family Pospiviroidae have single-stranded circular RNA genomes that adopt a rod-like or a quasi-rod-like conformation. These genomes contain a central conserved region that is involved in replication in the nucleus through an asymmetric RNA-RNA rolling-circle mechanism. Members of the family Pospiviroidae lack the hammerhead ribozymes that are typical of viroids classified in the family Avsunviroidae . The family Pospiviroidae includes the genera Apscaviroid , Cocadviroid , Coleviroid , Hostuviroid and Pospiviroid , with >25 species. This is a summary of the ICTV Report on the family Pospiviroidae , which is available at ictv.global/report/pospiviroidae.

The growth of the Tibetan Plateau throughout the past 66 million years has profoundly affected the Asian climate, but how this unparalleled orogenesis might have driven vegetation and plant diversity changes in eastern Asia is poorly understood. We approach this question by integrating modeling results and fossil data. We show that growth of north and northeastern Tibet affects vegetation and, crucially, plant diversity in eastern Asia by altering the monsoon system. This northern Tibetan orographic change induces a precipitation increase, especially in the dry (winter) season, resulting in a transition from deciduous broadleaf vegetation to evergreen broadleaf vegetation and plant diversity increases across southeastern Asia. Further quantifying the complexity of Tibetan orographic change is critical for understanding the finer details of Asian vegetation and plant diversity evolution., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

Iron deficiency anemia (IDA) is a common micronutrient deficiency among pregnant women with severe consequences including impaired immuno-inflammatory system, premature birth, fetal death etc. The present study aimed to investigate the effects of three iron supplements on IDA female rats and their offspring. The IDA female rat model was established with low iron diet and the rats were then mated. After pregnancy, rats were fed diets containing different iron supplements (iron polysaccharide complex, iron protein succinylate and ferrous sulfate) until their offspring were 42 days old. Pregnancy outcomes, haematological, iron metabolism, physical and neurological development indexes were determined. The results showed that all three iron supplements improved the levels of hematological parameters of both mother and offspring rats. After iron supplementation, serum iron, transferrin saturation and serum ferritin levels were increased compared with the IDA group. The level of ferritin light chain in the liver and spleen of both mother and offspring rats in iron supplemented groups was significantly higher than that of the IDA group. The average number of born alive per litter in the iron treatment groups was significantly higher than that in the IDA group. Iron supplements also improved the physical growth and neurobehavioral development of offspring rats. It was also found that iron supplementation improved the expression of ferritin light chain and the synaptic growth associated proteins in the brain and hippocampus. No significant difference was found in the efficacy of three iron supplements. These results suggest that pregnant and postpartum IDA affects pregnancy outcomes, offspring physical development and causes neural impairment. Sufficient iron supplementation can significantly improve IDA and its adverse effects on both mother and offspring.

Background: The Montessori Method underpinned by the principle of person-centered care has been widely adopted to design activities for people with dementia. However, the methodological quality of the existing evidence is fair. The objectives of this study are to examine the feasibility and effects of a culturally adapted group-based Montessori Method for Dementia program in Chinese community on engagement and affect in community-dwelling people with dementia., Methods: This was a two-arm randomized controlled trial. People who were aged 60 years or over and with mild to moderate dementia were recruited and randomly assigned to the intervention group to receive Montessori-based activities or the comparison group to receive conventional group activities over eight weeks. The attendance rates were recorded for evaluating the feasibility. The Menorah Park Engagement Scale and the Apparent Affect Rating Scale were used to assess the engagement and affect during the activities based on observations. Generalized Estimating Equation model was used to examine the intervention effect on the outcomes across the sessions., Results: A total of 108 people with dementia were recruited. The average attendance rate of the intervention group (81.5%) was higher than that of the comparison group (76.3%). There was a significant time-by-group intervention effect on constructive engagement in the first 10 minutes of the sessions (Wald χ 2  = 15.21-19.93, ps = 0.006-0.033), as well as on pleasure (Wald χ 2  = 25.37-25.73, ps ≤ 0.001) and interest (Wald χ2 = 19.14-21.11, ps = 0.004-0.008) in the first and the middle 10 minutes of the sessions, adjusted for cognitive functioning., Conclusions: This study provide evidence that Montessori-based group activities adapted to the local cultural context could effectively engage community-dwelling Chinese older people with mild to moderate dementia in social interactions and meaningful activities and significantly increase their positive affect., Trial Registration: ClinicalTrials.gov, NCT04352387. Registered 20 April 2020. Retrospectively registered.

Determining whether the high-latitude Bering land bridge (BLB) was ecologically suitable for the migration of mesothermal plants is significant for Holarctic phytogeographic inferences. Paleobotanical studies provide a critical source of data on the latitudinal positions of different plant lineages at different times, permitting assessment of the efficacy of the BLB for migration. Here we report exceptionally preserved fossils of Firmiana and Tilia endochrysea from the middle Miocene of South Korea. This represents a new reliable record of Firmiana and the first discovery of the T. endochrysea lineage in the fossil record of Asia. The occurrence of these fossils in South Korea indicates that the two lineages had a distribution that extended much farther north during the middle Miocene, but they were still geographically remote from the BLB. In light of the broader fossil record of Asia, our study shows that, in the middle Miocene, some mesothermal plants apparently inhabited the territory adjacent to the BLB and thus they were possibly capable of utilizing the BLB as a migratory corridor. Some other mesothermal plants, such as Firmiana and the T. endochrysea lineages, however, are restricted to more southern regions relative to the BLB based on current fossil evidence. These lineages may have been ecologically unable to traverse the BLB, which raises questions about the efficacy of the BLB as a universal exchange route for mesothermal plants between Asia and North America during the middle Miocene., Competing Interests: We have no competing interest., (© 2020 Kunming Institute of Botany, Chinese Academy of Sciences. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.)

Seven new diterpenoids (1-7), including five 7-membered ring vibsane-type diterpenoids, vibsanolide A-E (1-5) and a pair of epimers of 14,15,16,17-tetranorvibsane-type diterpenoids possessing bicyclo[4.2.1]nonane moiety, vibsanolide F-G (6-7), together with twelve known analogues (8-19) were isolated from the crude extracts of the leaves of Viburnum odoratissimum using Small Molecule Accurate Recognition Technology (SMART). These structures including absolute configurations were elucidated by means of comprehensive analyses of spectroscopic data, as well as comparison of the experimental and calculated electronic circular dichroism (ECD) spectra. These compounds were evaluated for their cytotoxic activities against A549 and HepG2 cells by MTT assay. The results showed that compound 2 exhibited potent cytotoxic activity against A549 cells with IC 50 value of 1.11 μM. Further staining experiments indicated that 2 could promote apoptosis induction, enhance reactive oxygen species (ROS) level and attenuate mitochondrial membrane potential (MMP) in A549 cells. Taken together, these findings provided new insights into understanding the cytotoxic activity of vibsane-type diterpenoids and it is meaningful to further investigate the application potential of V. odoratissimum., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Tibet's ancient topography and its role in climatic and biotic evolution remain speculative due to a paucity of quantitative surface-height measurements through time and space, and sparse fossil records. However, newly discovered fossils from a present elevation of ∼4,850 m in central Tibet improve substantially our knowledge of the ancient Tibetan environment. The 70 plant fossil taxa so far recovered include the first occurrences of several modern Asian lineages and represent a Middle Eocene (∼47 Mya) humid subtropical ecosystem. The fossils not only record the diverse composition of the ancient Tibetan biota, but also allow us to constrain the Middle Eocene land surface height in central Tibet to ∼1,500 ± 900 m, and quantify the prevailing thermal and hydrological regime. This "Shangri-La"-like ecosystem experienced monsoon seasonality with a mean annual temperature of ∼19 °C, and frosts were rare. It contained few Gondwanan taxa, yet was compositionally similar to contemporaneous floras in both North America and Europe. Our discovery quantifies a key part of Tibetan Paleogene topography and climate, and highlights the importance of Tibet in regard to the origin of modern Asian plant species and the evolution of global biodiversity., Competing Interests: The authors declare no competing interest., (Copyright © 2020 the Author(s). Published by PNAS.)

Emerging cellulose nanomaterials extracted from agricultural biomasses have recently received extensive attention due to diminishing fossil resources. To further reduce the carbon footprints and wastage of valuable resources, additive manufacturing techniques of new nanocellulosic materials have been developed. Studies on the preparation and characterization of 3D-printable functional nanocellulosic materials have facilitated a deeper understanding into their desirable attributes such as high surface area, biocompatibility, and ease of functionalization. In this critical review, we compare and highlight the different methods of extracting nanocellulose from biorenewable resources and the strategies for transforming the obtained nanocellulose into nanocomposites with high 3D printability. Optimistic technical applications of 3D-printed nanocellulose in biomedical, electronics, and environmental fields are finally described and evaluated for future perspectives., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Background: Mitochondrial transcription elongation factor (TEFM) is an essential molecule that regulates the replication-transcription switch of mitochondrial DNA. TEFM modulates both transcription elongation and RNA processing in mitochondria. The purpose of the present study was to determine the association of TEFM with tumor progression and prognosis in hepatocellular carcinoma (HCC) patients., Methods: The different protein expression level of TEFM among HCC cell lines was detected by Western blotting. The gene expression profiling interactive analysis (GEPIA) was used to dynamically analyze the mRNA expression of TEFM gene in different stages of HCC. The protein and mRNA expression levels of TEFM were detected by immunohistochemistry, Western blotting and qRT-PCR. The mRNA-SeqV2 expression of TEFM and clinical information of HCC patients were downloaded from the TCGA database by using R3.6.3 software. Next, the relationships between the expression level of TEFM and clinicopathological characteristics and the prognostic value of TEFM were analyzed. A Cox regression model was used for multivariate analysis of the factors that affected the prognosis of HCC. Finally, the association between the expression levels of TEFM and other mitochondrial regulatory genes and HCC biomarker genes was analyzed by GEPIA., Results: TEFM is upregulated in HCC cell lines compared to noncancerous liver cell line. TEFM protein and mRNA expression levels in HCC tissues were significantly upregulated compared with those in noncancerous liver tissues. In addition, the mRNA expression level of TEFM was significantly correlated with sex, serum AFP level, and vascular invasion (P<0.05). Further analysis showed that high expression level of TEFM was unfavorable in terms of the prognosis of patients with HCC. Cox multivariate regression analysis showed that patient age, vascular invasion, and TEFM expression were independent factors affecting the prognosis of HCC patients (P<0.05). The expression level of the TEFM gene was significantly positively correlated with the expression of multiple mitochondrial regulatory genes and biomarker genes of HCC (P<0.01, R>0)., Conclusions: Our findings reveal that TEFM may play an important role in the progression of HCC. More importantly, the elevated expression of TEFM may potentially predict poor overall survival (OS) and disease-free survival (DFS) in patients with HCC., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at: http://dx.doi.org/10.21037/jgo-20-120). The authors have no conflicts of interest to declare., (2020 Journal of Gastrointestinal Oncology. All rights reserved.)

Purpose: To perform a comprehensive meta-analysis of all available evidence on the efficacy and safety of catheter-based renal denervation for heart failure with reduced ejection fraction., Methods: We searched English and Chinese databases and calculated the weighted mean difference or standardized mean difference and 95% confidence intervals to estimate the efficacy and safety of renal denervation for heart failure. All relevant studies were screened and a meta-analysis was conducted using Review Manager 5.4., Results: A total of 11 studies were identified for the meta-analysis. For the primary outcomes, the results showed that renal denervation significantly improved ejection fraction (weighted mean difference 6.42), left ventricular end-systolic diameter (weighted mean difference -3.95), left ventricular end-diastolic diameter (weighted mean difference -4.17) and left atrial diameter (weighted mean difference -4.09). For the secondary outcomes, renal denervation reduced the B-type natriuretic peptide level, heart rate, systolic blood pressure and diastolic blood pressure. However, further analysis revealed that renal denervation improved heart function but did not further reduce the heart rate and blood pressure compared with the control group., Conclusion: Treatment with renal denervation can significantly improve heart function and structure in patients with heart failure. In addition, the level of B-type natriuretic peptide can be reduced after renal denervation treatment. Renal denervation did not further reduce heart rate and blood pressure compared with the control group. Therefore, the treatment of heart failure with renal denervation is effective and safe.

Background and Aims: Somatic mutation R249S in TP53 is highly common in hepatocellular carcinoma (HCC). We aim to investigate the effects of R249S in ctDNA on the prognosis of HCC., Methods: We analysed three cohorts including 895 HCC patients. TP53 mutation spectrum was examined by direct sequencing of genomic DNA from tissue specimens in HCC patients with hepatectomy (Cohort 1, N = 260). R249S and other recurrent missense mutations were assessed for their biological functions and associations with overall survival (OS) and progression-free survival (PFS) of HCC patients in Cohort 1. R249S within circulating tumour DNA (ctDNA) was detected through droplet digital polymerase chain reaction (ddPCR) and its association with OS and PRS was analysed in HCC patients with (Cohort 2, N = 275) or without (Cohort 3, N = 360) hepatectomy., Results: In Cohort 1, R249S occupied 60.28% of all TP53 mutations. Overexpression of R249S induced more serious malignant phenotypes than those of the other three identified TP53 recurrent missense mutations. Additionally, R249S, but not other missense mutations, was significantly associated with worse OS (P = .006) and PFS (P = .01) of HCC patients. Consistent with the results in Cohort 1, HCC patients in Cohorts 2 and 3 with R249S had worse OS (P = 8.291 × 10 -7 and 2.608 × 10 -7 in Cohorts 2 and 3, respectively) and PFS (P = 5.115 × 10 -7 and 5.900 × 10 -13 in Cohorts 2 and 3, respectively) compared to those without this mutation., Conclusions: TP53 R249S mutation in ctDNA may serve as a promising prognosis biomarker for HCC patients with or without hepatectomy., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Liver organoids have recently been applied as models for liver disease and drug screening, especially when combined with liver-on-a-chip technologies. Compared to hepatocyte-like cells, primary hepatocytes have high functionality but cannot maintain their function when cultured in vitro . Mesenchymal stem cells (MSCs) enhance hepatocyte function and maintain hepatocyte metabolism when co-cultured with hepatocytes. MSCs can help induced pluripotent stem cells to generate an organoid structure  via the MSC-based traction force triggered by extracellular matrix (ECM) proteins. In this study, primary hepatocytes were co-cultured with MSCs on a liver-derived ECM to generate liver organoids within a short duration., Aim: To create hepatocyte organoids by co-culturing primary hepatocytes with MSCs on a porcine liver extracellular matrix (PLECM) gel., Methods: Perfusion and enzymatic hydrolysis were used to form the PLECM gel. Rat hepatocytes and human MSCs were mixed and plated on pre-solidified PLECM gel in a 48-well plate for 48 h to generate organoids. Generated organoids were evaluated through hematoxylin and eosin, periodic acid-Schiff, immuno-histological, and immunofluorescence staining, and quantitative PCR for alb , CYP450 gene markers, and urea cycle genes. Culture medium was collected to detect albumin (ALB) and urea production on days 2, 4, 6, 8, 14, and 20., Results: The whole porcine liver was perfused and enzymatically hydrolyzed to form a PLECM gel. The structural components and basement membrane composition of the ECM, such as collagen type I, collagen type IV, fibronectin, and laminin, were demonstrated to be retained. Through interaction of human MSCs with the liver-derived ECM, primary hepatocytes and human MSCs assembled together into a 3D construction and generated primary hepatocyte organoids for 48 h. The mRNAs of the gene alb , the CYP450 gene markers cyp1a1, cyp1a2 , and cyp3a2 as well as urea cycle genes arg-1, asl, ass-1, cps-1, nags were highly expressed in hepatocyte organoids. Long-term survival of the primary hepatocyte organoids, as well as stable functionality, was demonstrated via ALB and urea production in vitro ., Conclusion: Our new method of creating primary hepatocyte organoids by co-culturing hepatocytes with MSCs on liver-derived ECM hydrogels could be used to develop models for liver disease and for drug screening., Competing Interests: Conflict-of-interest statement: All the authors have nothing to disclose., (©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.)

Sodium-ion batteries (SIBs) are heralded as promising candidates for grid-scale energy storage systems due to their low cost and abundant sodium resources. Excellent rate capacity and outstanding cycling stability are always the goals for SIBs. Up to now, nearly all attention has been focused on the control of morphology and structure of electrode materials, but the influence of binders on their performance is neglected, especially in cathode materials. Herein, using Na 3 V 2 (PO 4 ) 2 O 2 F (NVPOF) as a cathode material, the influence of four different binders (sodium alginate, SA; carboxymethylcellulose sodium, CMC; poly(vinylidene fluoride), PVDF; and poly(acrylic latex), LA133) on its electrochemical performance is studied. As a result, when using SA as the binder, the electrochemical performance of the NVPOF electrode is improved significantly, which is mainly because of the high water solubility, rich carboxyl and hydroxyl groups, and high adhesive and cohesive properties of the SA binder, leading to the uniform distribution of active materials NVPOF and carbon black in electrodes, good integrity, low polarization, and superior kinetic properties of the NVPOF electrodes, as demonstrated by scanning electron microscopy, cyclic voltammetry, electrochemical impedance spectroscopy, and galvanostatic intermittent titration technique. More importantly, when coupled with a hard carbon anode, the fabricated sodium-ion full cells also exhibit excellent rate performance, thus providing a preview of their practical application. This work shows that the battery performance can be improved by matching suitable binder systems, which is believed to have great importance for the further optimization of the electrochemical performance of SIBs.

Aldo-keto reductase KmAKR-catalyzed asymmetric reduction offers a green approach to produce dichiral diol tert-butyl 6-substituted-(3R,5R/S)-dihydroxyhexanoates, which are important building blocks of statins. In our previous work, we cloned a novel gene of NADPH-specific aldo-keto reductase KmAKR (WT) from a thermotolerant yeast Kluyveromyces marxianus ZJB14056 and a mutant KmAKR-W297H/Y296W/K29H (Variant III) has been constructed and displayed strict diastereoselectivity towards tert-butyl 6-cyano-(5R)-hydroxy-3-oxohexanoate ((5R)-1) but moderate activity and stability. Herein, to further co-evolve its activity and thermostability, we performed semi-rational engineering of Variant III by using a combinational screening strategy, consisting of tertiary structure analysis, loop engineering, and alanine scanning. As results, the "best" variant KmAKR-W297H/Y296W/K29H/Y28A/T63M (Variant VI) was acquired, whose K m , k cat /K m towards (5R)-1 was 0.66 mM and 210.77 s -1 mM -1 , respectively, with improved thermostability (half-life of 14.13 h at 40 °C). Combined with 1.5 g dry cell weight (DCW) L -1 Exiguobacterium sibiricum glucose dehydrogenase (EsGDH) for NADPH regeneration, 4.5 g DCW L -1 Variant VI completely reduced (5R)-1 of up to 450 g L -1 within 7.0 h at 40 °C, yielding the corresponding optically pure tert-butyl 6-cyano-(3R,5R)-dihydroxyhexanoate ((3R,5R)-3, >99.5% d.e. p ) with a space-time yield (STY) of 1.24 kg L -1 day -1 , and this was the highest level documented in literatures so far on substrate loading and STY of producing (3R,5R)-3. Besides (5R)-1, Variant VI displayed strong activity on tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate ((5S)-2). 4.5 g DCW L -1 Variant VI completely reduced 400 g L -1 (5S)-2, within 5.0 h at 40 °C, yielding optically pure tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate ((3R,5S)-4, >99.5% d.e. p ) with a STY of 1.34 kg L -1 day -1 . In summary, Variant VI displayed industrial application potential in statins biomanufacturing., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Avibacterium paragallinarum , the causative agent of infectious coryza, is known to release outer membrane vesicles (OMVs). In the present study, we investigated the composition, bioactivities, and functional properties of the OMVs of A. paragallinarum . Following extraction and purification, the OMVs were observed to be spherical in shape, with diameters ranging from 20 to 300 nm. The vesicles contained endotoxin as well as genomic DNA. The molecular weights of the OMV-contained protein fragments were mostly concentrated at 65 and 15 kDa. The components of the OMV proteins were mainly various functional enzymes (e.g., ATP-dependent RNA helicase), structural components (e.g., streptomycin B receptor and membrane protein), and some hypothetical proteins with unknown functions. The expression levels of inflammation-related factors, such as interleukin (IL)-2, IL-6, IL-1β, IL-10, and inducible nitric oxide synthase (iNOs), were significantly upregulated in chicken macrophage cells HD11 incubated with OMVs. Serum IgG antibodies were measured after two intramuscular injections of an OMV-based vaccine into specific pathogen-free (SPF) chickens. The vaccinated chickens were then challenged by A. paragallinarum of homologous and heterologous serovars. It was noted that the vaccinated chickens produced immunoglobulin G (IgG) antibodies against A. paragallinarum . The OMVs conferred an acceptable level of protection (70%), defined as an absence of colonization and of clinical signs, against the homologous strain (serovar A), while the cross-protection against heterologous challenge with serovars B and C was much weaker. However, the OMVS did provide significant protection against clinical signs for all three serovars. Overall, this study laid a foundation for further unraveling the functional roles of OMVs released by A. paragallinarum ., (Copyright © 2020 Mei, Sun, Blackall, Xian, Li, Gong and Wang.)

Metal-organic frameworks (MOFs) with cuprous-halide-aggregates have shown superiority as organic LED (OLED) and semiconductor materials, while engineering MOF flexibility by involving the expansion of cuprous aggregates remains a great challenge. In this particular work, a dissolution-recrystallization structural transformation (DRST) with the dramatic growth of Cu I -I aggregates, from 2D NJNU-100 to 3D NJNU-101 has been successfully realized. The unsaturated coordination nodes (2-positional nitrogen atoms) in NJNU-100 have been demonstrated to be the driven force for DRST to NJNU-101 via the formation of coordination bonds. The structural transformation process was irreversible and observed with optical microscopy and powder XRD. The expansion of Cu I -I aggregates was also computational simulated accompanying with the rotation of the neutral tripodal TTTMB ligand (1,3,5-tris(1,2,4-triazol-1-ylmethyl)-2,4,6-trimethylbenzene) and the reduction of Cu II to Cu I . Moreover, the intermediate product NJNU-102 was captured by adding the planar molecular anthrancene to shut down the reaction, where only partial 2-positional nitrogen atoms coordinated to the aggregates and the anthrancene was oxidized to anthraquinone. NJNU-102 has further confirmed that DRST involved the breakage and recombination of coordination bonds and the electron transfer. NJNU-100 and NJNU-101 could be applied as semiconductor and OLED materials. This work has provided insights for crystal engineering, especially for the construction of the Cu I x X y aggregates, and illustrated that DRST could be controlled with a rational design (as the unsaturated coordination modes).

Exosomes contain a variety of biological active substances such as proteins, miRNAs, lncRNAs and lipids, and exosomes from different cells play different biological functions. Exosomes, as a carrier, are involved in many pathological processes such as nerve injury and repair, vascular regeneration, immune response, and fibrosis formation. It plays an important role in the treatment of eye diseases such as glaucoma, diabetic retinopathy, and keratitis. This paper reviews the research progress of exosomes in various diseases in vivo , which provides a new way for the treatment of eye diseases., (International Journal of Ophthalmology Press.)

Forsythiae Fructus is divided into Qingqiao and Laoqiao due to different harvesting periods. So far, the accumulation of heavy metals in the two types of Forsythiae Fructus has not been reported. In this study, the residual levels of copper(Cu), lead(Pb), chromium(Cr), arsenic(As), cadmium(Cd) and mercury(Hg) in 29 batches of Laoqiao and 60 batches of Qingqiao were determined by inductively coupled plasma mass spectrometry(ICP-MS). The samples were collected from Shanxi, Shaanxi, Henan, and Hebei Provinces. In addition, the diversity and correlation of harmful elements in Qingqiao and Laoqiao were analyzed by multivariate statistical method. Furthermore, principal component analysis(PCA) was used to analyze the harmful elements concentrations of Qingqiao and Laoqiao. The results showed that there was a significant difference on the residual levels of heavy metals and harmful elements between Qingqiao and Laoqiao. Among them, the content of Pb in Laoqiao is significantly higher than that in Qingqiao(P&lt;0.01), while the content of Cu is significantly lower than that in Qingqiao. However, the difference in harmful elements among different producing areas of Forsythiae Fructus is not significant. PCA analysis showed that Qingqiao and Laoqiao were successfully grouped into two categories. This study suggests significant difference in the residual levels of heavy metals and harmful elements between Qingqiao and Laoqiao. Besides, Forsythiae Fructus has a certain enrichment of Pb in the fruit ripening stage(Laoqiao). This study provides a reference for the quality classification and safety of Forsythiae Fructus.

Salvia plebeia has been in use as traditional Chinese medicine (TCM) for more than 500 years. In this study, the complete chloroplast (cp) genome of S. plebeia was sequenced, assembled and compared to those of other five published Salvia cp genomes. It was found that the cp genome structure of S. plebeia was well conserved and had a total size of 151 062 bp. Four parameters were used to display the usage conditions of the codons of the amino acids in Salvia genus. Although the number of protein-coding genes in each species was the same, the total number of codons was different. Except for amino acids Trp and Met whose Relative Synonymous Codon Usage (RSCU) value of one condon was equal to 1, the remaining 19 amino acids had 1-3 preferred codons. The preferred codon names of each amino acid were coincident. The period size for the tandem repeats of six species ranged from 9 to 410 bp. Salvia cp genomes mainly possessed tandem repeats with a copy number less than or equal to 3. The sequence length of tandem repeats of the six species ranged from 25 to 824 bp. Highly viarable regions including four intergenic spacers and six partial genes were discovered as potential specific barcodes for Salvia species through cp genome-wide comparison. Finally, we performed phylogenetic analyses based on the complete cp genome and coding sequences respectively. These results provide information to help construct the cp genome library for Salvia, which may support studies of phylogenetics, DNA barcoding, population and transplastomics., (Copyright © 2020 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.)

Background: This report serves to describe the use of a novel adjustable single 8-0 polypropylene suture for scleral fixation without conjunctival dissection, and to describe related clinical outcomes associated with this approach., Methods: In this study, we retrospectively reviewed 28 eyes from 27 patients that underwent scleral fixation of the intraocular lens (IOL) without conjunctival dissection using an adjustable single 8-0 polypropylene suture at the Beijing Tongren Eye Center between April 2018 and April 2019. For this surgical approach, a 23-gauge infusion cannula was set, after which two Hoffmann scleral pockets were created. Next, 8-0 polypropylene sutures were inserted into the eye guided by 10-0 polypropylene sutures of a long straight needle. The 8-0 suture was then used to fix the haptic IOs. Finally, these 8-0 polypropylene sutures were removed from the scleral pockets, and knots were tightened with the adjustable single suture. Primary outcomes included visual acuity and postoperative complication incidence., Results: For this study, outcomes for 28 eyes from 27 patients (9 female, 18 male) were assessed. Patients had a mean age of 54 ± 15.11 years-old and were followed for an average of 10.18 ± 2.76 months postoperatively. Uncorrected visual acuity in these patients improved significantly from a preoperative value of 1.269 ± 0.464 logMAR to a 3-month postoperative value of 0.409 ± 0.413 logMAR (p = 0.000). The majority of postoperative complications in these patients were temporary and self-limiting, including corneal edema (35.71%), hypotony (14.29%), elevated intraocular pressure (28.58%), and mild hyphema (7.14%). No evidence of exposure or erosion of the trimmed suture end was detected in any patients. An ultrasound biomicroscope was able to readily detect the IOL and all sutures, and IOLs were found to be well-centered without any dislocation, tilting, or subluxation upon follow-up., Conclusions: An adjustable single 8-0 polypropylene suture can reliably and effectively be used for scleral fixation without conjunctival dissection for the treatment of patients with aphakia or inadequate posterior capsule support. The novel procedure described herein may therefore be an effective means of minimizing the risk of suture-related complications in patients undergoing scleral-fixated IOL implantation., Trial Registration: Retrospective case series study, not applicable. NCT04476264 .