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2-Phenylethanol (2-PE), a higher alcohol with a rose-like odor, has been widely utilized in food, perfume, and beverages. Saccharomyces cerevisiae is one of the most promising microorganisms for the biosynthesis of natural 2-PE. However, the growth of S. cerevisiae is generally inhibited by 2-PE, which makes its production in yeast cell factories challenging. Here, the whole-cell bioconversion was used to avert growth inhibition, leading to an increase in the concentration and productivity of 2-PE. Moreover, rapamycin (Rap) addition further improved the efficiency of 2-PE synthesis. The concentration of 2-PE (2.20 g/L) was 1.68-fold higher than that in the absence of Rap during the whole-cell bioconversion by S. cerevisiae BY4741. RT-qPCR results showed that Rap addition increased the transcription of ARO9, ARO10, ADH2, GAP1, ARO80, GLN3, and GDH2. When the GLN3 was knocked out, the transcriptional levels of the genes were dramatically decreased, and the concentration of 2-PE significantly decreased to 0.21 g/L. The results indicated that Rap enhanced the flux of the Ehrlich pathway, and Gln3 exerted a central role in the regulation of Rap. Furthermore, commercial yeast (S. cerevisiae FY202001) was selected to verify the applicability of Rap. In the presence of Rap, 3.67 g/L 2-PE was obtained by whole-cell bioconversion in flask, which was increased by 9% than that in the absence of Rap. Finally, the 2-PE titer reached 4.93 g/L by whole-cell bioconversion in a 5 L bioreactor, with a yield of 84 mol% from L-phenylalanine and a productivity of 0.103 g/L h, which was far higher than that of the currently reported in S. cerevisiae. These findings provided a new idea for the efficient synthesis of 2-PE. KEY POINTS: • Whole-cell bioconversion was used to produce 2-PE. • The regulation of the Ehrlich pathway by Rap provides a theoretical basis for developing an effective yeast cell factory to produce 2-PE. • The 2-PE productivity of 0.103 g/L h is far higher than that of the currently reported in S. cerevisiae .

Two yellow-pigmented bacterial strains, LZ-14

Background To estimate the incidence, prevalence and incidence-based mortality in patients with gynecologic sarcoma (GS), and described the trends of survival and initial treatments in the US by using the Surveillance, Epidemiology, and End Results (SEER) database. Methods GS cases aged 20 years or older between 1975 and 2015 were identified from SEER 9 registries. Incidence, prevalence, and incidence-based mortality were estimated, all rates were age adjusted to the 2000 US standard population and presented as per 100,000 persons. Annual percentage change (APC) and average APC (AAPC) were calculated to describe the trends. In addition, stage distribution, cancer-specific survival (CSS) and initial treatment pattern over time were also reported. Results The overall age-adjusted incidence of GS increased from 2.38 to 3.41 per 100,000 persons from 1975 to 2015, with an AAPC of 1.0 (P P Conclusions The incidence of GS increased significantly with improved survival, which might due to the strategy of combination of multiple treatment. However, no obvious improvement on the early detection of GS was found, which should be facilitated to further improve the prognosis of GS.

Objective: The objective of the study was to investigate the effect of multiple clinical parameters (age, weight, blood types, and bleeding types) on FVIII pharmacokinetic parameters (PK parameters) in adult patients with severe hemophilia A (SHA), draw up individualized preventive treatment plans, and observe clinical efficacy. Methods: Forty SHA patients treated in our hospital from January 2018 to May 2019 were enrolled, with their age, weight, blood types, bleeding types, and PK parameters measured to analyze the effects of clinical parameters on PK parameters. Individualized preventive treatment was developed, and patients were followed up for 1 year. The annual bleeding rate (ABR), annual joint bleeding rate (AJBR), and annual FVIII dosage were observed and compared before and after treatment. Results: Weight, blood types, and bleeding types could affect the PK parameters of FVIII. A prophylaxis plan was formulated under the guidance of FVIII half-life. After 1 year of follow-up, the mean ABR dropped from 36.54 to 4.06, decreased by 88.9%, the mean AJBR dropped from 28.36 to 2.75, decreased by 90%, and annual FVIII dosage increased by 47%. The dosage of FVIII in 8 patients after was less than that before prophylaxis, and the average half-life time of these 8 patients was 13.32 h. Conclusions: (1) Weight, blood types, and bleeding types of adult SHA patients could affect FVIII half-life. As body mass index increased, FVIII half-life was significantly prolonged. The FVIII half-life of patients with type O blood was significantly shorter than those with other blood types, and the FVIII half-life of knee joint bleeding was conspicuously shorter than those of elbow joint bleeding. (2) Individualized preventive treatment could markedly reduce the number of bleeds. For patients with a long half-life period, the total annual FVIII dosage could be reduced to achieve bleeding prevention.

Background Pulmonary endarterectomy (PEA) is the preferred treatment for CTEPH patients which can significantly improve symptoms and pulmonary hemodynamics. Therefore, this retrospective study evaluated the long-term outcomes after pulmonary endarterectomy (PEA) and analyze the predictors of long-term outcomes for chronic thromboembolic pulmonary hypertension (CTEPH). Methods From 2002–2020, 76 CTEPH patients successfully discharged after PEA in Beijing Chaoyang Hospital were followed-up by scheduled clinical visits or telephone interviews. The follow-up time lasted for 18 years and median time was 7.29 years. Results The survival rate at 1,3,5,10,15 years postoperatively was 100.00%, 97.10%, 95.40%, 89.80% and 82.90%, respectively. Multivariate logistics regression analysis showed that postoperative mPAP (hazard ratio: 1.144; 95%confidence interval: 1.018–1.285; P = 0.023) was associated with a higher risk of late death, right atrium right and left diameters (hazard ratio: 1.113; 95%confidence interval, 1.006–1.231; P = 0.038) were associated with a higher risk of major adverse events. Conclusion Pulmonary endarterectomy is an effective way to treat CTEPH. Long-term outcome is excellent for patients who undergoing pulmonary endarterectomy who survived from peri-operation time. Postoperative mPAP is a significant prognostic factor for long-term death and right atrium right and left diameters is a significant prognostic factor for major adverse events. That shows patients with high postoperative mPAP and right atrium right and left diameter should be followed up closely.

BACKGROUND Colorectal cancer (CRC) is one of the most common malignant tumors of the digestive tract. Lymphatic metastases of this tumor are mostly confined to the regional lymph nodes, and distant supraclavicular lymph node metastases are very rare. CASE SUMMARY In this report, we describe a patient with sigmoid carcinoma and isolated synchronous supraclavicular lymph node metastases. A 56-year-old male presented with a left cervical mass that was confirmed as a lymph node metastasis from sigmoid cancer by several auxiliary examinations. After 6 cycles of chemotherapy with the 5-fluorouracil, leucovorin and oxaliplatin + cetuximab regimen, the sigmoid colon tumor and Virchow’s lymph node metastasis were significantly smaller than before treatment, and no new metastatic sites were observed. Considering the effects of chemotherapy on quality of life, resection of the primary tumor was performed followed by 4 cycles of chemotherapy with the original chemotherapy regimen. Virchow’s lymph node dissection was selected by mutual consultation between the patient and us. After the second surgery, the patient received capecitabine and cetuximab chemotherapy and did not experience recurrence or metastasis during follow-up. CONCLUSION In conclusion, supraclavicular lymph node metastasis without any other solid organ metastasis is a potential metastatic pathway for CRC. In addition, after resection of the primary lesion, postoperative chemotherapy combined with supraclavicular lymph node dissection is feasible for the treatment of patients with CRC and isolated synchronous Virchow’s lymph node metastases.

A novel actinobacterium NJES-13T was isolated from the gut of Antarctic emperor penguin Aptenodytes forsteri. The new isolate produces bioactive gephyromycin metabolites and exopolysaccharides (EPS). Cells were Gram-negative, motile with the peritrichous flagella, and with a faint layer of extracellular slime. Colonies were yellow when grown on marine agar, ISP1, 2, 4 and TSA media. The strain developed clusters of coccoid, and divided by binary fission in the early phase of growth. The cell clusters were gradually disrupted during the stationary phase and formed short rod-shape cells which were interconnected by viscous EPS showing a three-dimensional net-like morphology, and contained polyhydroxyalkanoates (PHA) granules inside the cells. Growth of strain NJES-13T was observed at 15–45 °C, at pH 6.0–9.0 with 0.5–9.0% (w/v) NaCl. The complete genomic size of strain NJES-13T was 3.45 Mb with a DNA G + C content of 67.0 mol%. The combined polyphasic taxonomic characterizations presented in this study unequivocally separated strain NJES-13T from all known genera in the family Dermatophilaceae. Thus, strain NJES-13T represents a novel species of a new genus, for which the name Gephyromycinifex aptenodytis gen. nov., and sp. nov. is proposed. The type strain is NJES-13T (= CCTCC 2019007T = KCTC 49281T). Genetic prediction of secondary metabolite biosynthesis revealed a 44.5 kb-long biosynthetic gene cluster (BGC) of type III polyketide synthase (PKS) as well as four other BGCs, indicating its great potential to produce novel bioactive metabolites derived from the gut microbiota of animals living in the extreme habitats in the Antarctica.

Marine phycosphere hosts cross-kingdom algae-bacteria interactions playing a variety of crucial roles in aquatic ecosystems especially for the prevention and control of harmful algal blooms (HABs). During the investigation of structural composition of phycosphere microbiota (PM) of diverse marine HAB dinoflagellates, a Gram-negative, strictly aerobic, non-motile and rod-shaped bacterium designated LZ-17T was isolated from the phycosphere of highly toxic Alexandrium catenella LZT09. The 16S rRNA gene sequence analysis and the multilocus sequence analysis (MLSA) based on five protein-coding housekeeping genes (atpD, gyrB, mutL, topA and rpoD) indicated that strain LZ-17T was affiliated to the genus Maritimibacter within the family Rhodobacteraceae, and closely related to Maritimibacter alkaliphilus HTCC2654T (99.1%), ‘Maritimibacter harenae’ DP07T (97.9%) and M. lacisalsi X12M-4T (95.7%). The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain LZ-17T and the type strain of M. alkaliphilus were 96.9% and 74.7%. However, strain LZ-17T could be clearly distinguished from its closest by the phenotypical and phenotypical characteristics. Strain LZ-17T contained Q-10 as its major isoprenoid quinone, and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c), C16:0 and C16:0 2-OH as the predominant fatty acids (>10%). The major polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylcholine. The DNA G + C content was 64.3 mol%. Based on the polyphasic taxonomic characterization, strain LZ-17T represents a novel species of the genus Maritimibacter, for which the name Maritimibacter alexandrii sp. nov. is proposed, with the type strain LZ-17T (=CCTCC 2019005T = KCTC 72193T).

Alzheimer's disease (AD) is a prevalent type of dementia and threatens the health of most elderly people and poses a huge burden to families and society. The fibroblast growth factor 23 (FGF23)/α-Klotho axis is associated with multiple aging-related diseases. Hence, this study explored the mechanism of the FGF23/α-Klotho axis in AD. FGF23/α-Klotho protein contents and levels of inflammatory cytokines in AD patients were measured, and the correlation between FGF23/α-Klotho protein contents and inflammatory cytokines was analyzed. FGF23 and α-Klotho expressions were blocked in peripheral blood mononuclear cells (PBMCs) in AD patients (AD-PBMCs) to assess the effects on cell inflammation and the Wnt/β-catenin pathway activation. The Wnt/β-catenin pathway was inhibited to evaluate cell inflammation. Combined treatments of the cells were conducted to verify the role of the FGF23/α-Klotho axis and the Wnt/β-catenin pathway in inflammation in AD-PBMCs. Increased FGF23 protein concentration and reduced α-Klotho protein concentration were observed in AD patients and correlated with inflammatory cytokine levels. FGF23 inhibition or α-Klotho overexpression reduced the production of inflammatory cytokines and activated the Wnt/β-catenin pathway in AD-PBMCs. Blocking the Wnt/β-catenin pathway increased inflammatory cytokine production in AD-PBMCs and annulled the effects of the FGF23/α-Klotho axis on AD-induced cell inflammation. We concluded that the FGF23/α-Klotho axis can regulate the AD-induced cell inflammation through the Wnt/β-catenin pathway.

In this study, a novel method of coupling phytohormones with saline wastewater was proposed to drive efficient microalgal lipid production. All the six phytohormones effectively promoted microalgae growth in saline wastewater, and further increased the microalgal lipid content based on salt stress, so as to achieve a large increase in microalgal lipid productivity. Among the phytohormones used, abscisic acid had the most significant promoting effect. Under the synergistic effect of 20 g/L salt and 20 mg/L abscisic acid, the microalgal lipid productivity reached 3.7 times that of the control. Transcriptome analysis showed that differentially expressed genes (DEGs) of microalgae in saline wastewater were mainly up-regulated under the effects of phytohormones except brassinolide. Common DEGs analysis showed that phytohormones all regulated the expression of genes related to DNA repair and substance synthesis. In conclusion, synergistic effect of salt stress and phytohormones can greatly improve the microalgal lipid production efficiency.

The nomenclature of duck viral hepatitis (DVH) was historically not a problem. However, 14 hepatotropic viruses among 10 different genera are associated with the same disease name, DVH. Therefore, the disease name increasingly lacks clarity and may no longer fit the scientific description of the disease. Because one disease should not be attributed to 10 genera of viruses, this almost certainly causes misunderstanding regarding the disease-virus relationship. Herein, we revisited the problem and proposed an update to DVH disease classification. This classification is based on the nomenclature of human viral hepatitis and the key principle of Koch's postulates (“one microbe and one disease”). In total, 10 types of disease names have been proposed. These names were literately matched with hepatitis-related viruses. We envision that this intuitive nomenclature system will facilitate scientific communication and consistent interpretation in this field, especially in the Asian veterinary community, where these diseases are most commonly reported.

Aims Vibrio (V.) parahaemolyticus is one of the most frequently occurred pathogens in mariculture. This study aimed to explore the mechanism of the impact of Ulva fasciata (U. fasciata) on the motility and biofilm formation of V. parahaemolyticus. Methods and results The inhibitory effect of U. fasciata on a V. parahaemolyticus, isolated from clam maricultural sediment, was examined by co-culture. The live U. fasciata significantly inhibited the swimming behaviour, twitching behaviour and biofilm formation of V. parahaemolyticus JF, with inhibition rates ranging from 2.48%-20.26%, 1.59-39.18%, and 28.3-94.7% under different nitrate and phosphate levels. The results of transcriptome sequencing showed that 210 significantly differentially expressed genes were found in strain JF between the presence and absence of U. fasciata, including 90 upregulated genes and 120 downregulated genes. According to GO (Gene Ontology) function enrichment and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis, the downregulated genes of JF were partially enriched in flagella assembly (fliC, fliK, fliG, fliN, fliH, fliI, fliJ, fliA), bacterial chemotaxis (mCP, cheB, cheW, cheY) and biofilm formation (fliA/σ28 , eps), which explained the suppressed motility and biofilm formation of V. parahaemolyticus JF under U. fasciata stress. Conclusions Live U. fasciata significantly impaired the motility and biofilm formation of V. parahaemolyticus, which could occur in niches with either sufficient or inadequate nutrient (nitrate and phosphate) concentrations. The differentially expressed genes of V. parahaemolyticus modulated by U. fasciata were enriched mainly in the flagellar assembly, bacterial chemotaxis and biofilm pathways. Significance and impact of study New information on how V. parahaemolyticus respond to U. fasciata regarding motility and adhesion behaviours, and the mechanism of that was firstly explored in this study. The results suggested that the seaweed U. fasciata has promising prospects as an environmentally friendly preventive measure to treat vibriosis in mariculture.

Tumor necrosis factor receptor 1 (TNFR1) associated death domain protein (TRADD) is a pivotal adaptor in TNF signaling pathway and plays an important role in apoptosis and immune regulation. The function of TRADD has been investigated extensively in mammals, however, the role of TRADD in ducks remains obscure. To reveal the function of duck TRADD (duTRADD) in the apoptosis and innate immune response, the TRADD homologue of mallard (Anas platyrhynchos) has been cloned and the function of duTRADD is investigated in this study. We conducted sequence analysis of the duTRADD, the open reading frame (ORF) region of duTRADD gene was 1065 bp, encoding 354 amino acids (aa), which shares similar functional domain to its mammalian counterpart. Tissue distribution profile of duTRADD in 7-day-old ducklings showed that the expression level of the gene was the highest in heart, followed by liver and brain. Accordingly, duck Tembusu virus (DTMUV) has been shown to decrease duTRADD expression, while overexpression of duTRADD inhibited DTMUV replication in a dose-dependent manner. Furthermore, duTRADD activated the transcriptional activity of caspase-3/8/9, the flow cytometry showed that duTRADD significantly induced apoptosis. However, duTRADD showed hardly any effect on the transcriptional activity of IFN-α/β and its downstream interferon-stimulated genes (ISGs). The current data support the conclusion that duTRADD is a novel pro-apoptotic protein with a critical role in defense against DTMUV invasion. These results lay the theoretical foundation for the development of new anti-DTMUV strategies.

Highly pathogenic coronaviruses, including SARS-CoV-2, SARS-CoV and MERS-CoV, are thought to be transmitted from bats to humans, but the viral genetic signatures that contribute to bat-to-human transmission remain largely obscure. In this study, we identified an identical ribosomal frameshift motif among the three bat–human pairs of viruses and strong purifying selection after jumping from bats to humans. This represents genetic signatures of coronaviruses that are related to bat-to-human transmission. To further trace the early human-to-human transmission of SARS-CoV-2 in North America, a geographically stratified genome-wide association study (North American isolates and the remaining isolates) and a retrospective study were conducted. We determined that the single nucleotide polymorphisms (SNPs) 1,059.C > T and 25,563.G > T were significantly associated with approximately half of the North American SARS-CoV-2 isolates that accumulated largely during March 2020. Retrospectively tracing isolates with these two SNPs was used to reconstruct the early, reliable transmission history of North American SARS-CoV-2, and European isolates (February 26, 2020) showed transmission 3 days earlier than North American isolates and 17 days earlier than Asian isolates. Collectively, we identified the genetic signatures of the three pairs of coronaviruses and reconstructed an early transmission history of North American SARS-CoV-2. We envision that these genetic signatures are possibly diagnosable and predic markers for public health surveillance.

The fall armyworm (Spodoptera frugiperda) is one of the most significant agricultural pests in the world and invaded China in early 2019. We sampled and sequenced RNA-seq data from 15 individuals across different developmental stages. Developmental stages were the larval stage (5th instar larvae and 6th instar larvae), chrysalis stage, and adult stage (female adult and male adult). Individual samples were mainly clustered by developmental stages and we then identified variation between developmental stages of differentially expressed transcripts (DETs). There were 2136 upregulated DETs and 1391 downregulated DETs in the larval stage when comparing larval and chrysalis stages. In the comparison between the chrysalis and adult stages, there were 2033 upregulated DETs and 1391 downregulated DETs in the chrysalis stage. In total, 19,195 abundantly expressed transcripts were obtained and 10% of them were DETs. We then obtained stage-specific DETs to investigate the potential function of the fall armyworm during different developmental stages. We also constructed our annotation background set for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. This indicated that the fall armyworm may undergo active metabolism during its lifespan, even in the chrysalis stage. And it also may experience detoxifying and xenobiotic metabolism throughout its life, especially in the larval stage, which partially explains the difficulty to eradicate using chemical control. Our study is the first insight into the developmental patterns of the fall armyworm and we also provide the fundamental information about enhanced drug resistance at the level of transcriptome. These results are beneficial for a future investigation related to the eradication and/or control stage.

Riemerella anatipestifer (R. anatipestifer, RA) is an infectious pathogen that causes septicemia and polyserositis in ducks. Our previous studies showed that RA CH-1 ∆fur was significantly attenuated in ducklings, which highlights the potential of this strain as a live attenuated vaccine. In this study, it was shown that infection with 10

Phytochemical investigation on the 80% EtOH extract of the whole plants of Zephyranthes grandiflora afforded three new 4a-epi-plicamine-type alkaloids, zephyranthines A-C (1-3). Structural elucidat...

Duck enteritis virus (DEV) multifunctional tegument protein UL13 is predicted to be a conserved herpesvirus protein kinase; however, little is known about its subcellular localization signal. In this study, through transfection of 2 predicted nuclear signals of DEV UL13 fused to enhanced green fluorescent protein, 2 bipartite nuclear localization signals (NLS) were identified. We found that ivermectin blocked the NLS-mediated nuclear import of DEV UL13, showing that the nuclear localization signal of DEV UL13 is a classical importin α- and β-dependent process. We constructed a DEV UL13 mutant strain in which the NLS of DEV UL13 was deleted to explore whether deletion of the NLS affects viral replication. Amino acids 4 to 7 and 90 to 96 were predicted to be NLSs, further proving that nuclear import occurs via a classical importin α- and β-dependent process. We also found that the NLS of pUL13 had no effect on DEV replication in cell culture. Our study enhances the understanding of DEV pUL13. Taken together, these results provide significant information regarding the biological function of pUL13 during DEV infection.

To determine the role of glycoprotein I (gI) in duck plague virus (DPV), a gI-deleted mutant (BAC-CHv-ΔgI) and a gI-revertant virus (BAC-CHv-ΔgI Rev) were constructed by using a markerless two-step Red recombination system implemented on the DPV genome cloned into a bacterial artificial chromosome (BAC). Mutants were characterized on duck embryo fibroblast (DEF) cells compared with wild-type virus. BAC-CHv-ΔgI produced viral plaques on DEF cells that were on average approximately 57.2% smaller than those produced by BAC-CHv-ΔgI Rev and wild-type virus. Electron microscopy confirmed that deleting of gI resulted in nucleocapsids accumulated around the cytoplasm vesicles and few of them could complete the final envelopment process. These results clearly indicated that DPV gI plays significant roles in viral cell-cell spread and viral final envelopment process.

Viscoelastic hydrogels can enhance 3D cell migration and proliferation due to the faster stress relaxation promoting the arrangement of the cellular microenvironment. However, most synthetic photocurable hydrogels used as bioink materials for 3D bioprinting are typically elastic. Developing a photocurable hydrogel bioink with fast stress relaxation would be beneficial for 3D bioprinting engineered 3D skeletal muscles in vitro and repairing volumetric muscle loss (VML) in vivo; however, this remains an ongoing challenge. This study aims to develop an interpenetrating network (IPN) hydrogel with tunable stress relaxation using a combination of gelatin methacryloyl (GelMA) and fibrinogen. These IPN hydrogels with faster stress relaxation showed higher 3D cellular proliferation and better differentiation. A 3D anisotropic biomimetic scaffold was further developed via a printing gel-in-gel strategy, where the extrusion printing of cell-laden viscoelastic FG hydrogel within Carbopol supported gel. The 3D engineered skeletal muscle tissue was further developed via 3D aligned myotube formation and contraction. Furthermore, the cell-free 3D printed scaffold was implanted into a rat VML model, and both the short and long-term repair results demonstrated its ability to enhance functional skeletal muscle tissue regeneration. These data suggest that such viscoelastic hydrogel provided a suitable 3D microenvironment for enhancing 3D myogenic differentiation, and the 3D bioprinted anisotropic structure provided a 3D macroenvironment for myotube organization, which indicated the potential in skeletal muscle engineering and VML regeneration. STATEMENT OF SIGNIFICANCE: The development of a viscoelastic 3D aligned biomimetic skeletal muscle scaffold has been focused on skeletal muscle regeneration. However, a credible technique combining viscoelastic hydrogel and printing gel-in-gel strategy for fabricating skeletal muscle tissue was rarely reported. Therefore, in this study, we present an interpenetrating network (IPN) hydrogel with fast stress relaxation for 3D bioprinting engineered skeletal muscle via a printing gel-in-gel strategy. Such IPN hydrogels with tunable fast stress relaxation resulted in high 3D cellular proliferation and adequate differentiation in vitro. Besides, the 3D hydrogel-based scaffolds also enhance functional skeletal muscle regeneration in situ. We believe that this study provides several notable advances in tissue engineering that can be potentially used for skeletal muscle injury treatment in clinical.

Acute lung injury (ALI) is a common complication in patients with sepsis and is accompanied by high mortality. The present study aimed to investigate if the organic compound citrulline has a protective against lipopolysaccharide (LPS)-stimulated ALI and its potential mechanisms. ALI was induced in mice by intraperitoneal (i.p.) injection of LPS (10 mg/kg). Citrulline (1 g/kg/day) was administrated i.p. 7 days prior to LPS injection. Mouse lung vascular endothelial cells (MLVECs) were divided into five groups: Control, LPS, LPS + Cit, LPS + N-acetyl-L-cysteine (NAC) and LPS + Cit + ML385. Lung injury was determined by morphology changes. Apoptosis and pyroptosis were detected using western blot analysis and immunofluorescence. The present results indicated that citrulline can significantly attenuate ALI. Citrulline pretreatment decreased the expression of NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome and decreased pyroptosis and apoptosis. Intervention with the total reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine attenuated NLRP3 inflammasome-associated pyroptosis and apoptosis in LPS-treated MLVECs. Citrulline pretreatment inhibited pyroptotic cell death and apoptosis induced by LPS. Citrulline decreased accumulation of intracellular ROS and activated the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. Furthermore, the Nrf2 inhibitor ML385 reversed ROS generation, NLRP3 inflammasome-mediated pyroptosis and apoptosis suppressed by citrulline. In summary, the present data demonstrated that citrulline may confer protection against ALI via inhibition of ROS/NLRP3 inflammasome-dependent pyroptosis and apoptosis via the Nrf2 signaling pathway.

A novel Gram-negative, aerobic, motile and short rod-shaped bacterium with exopolysaccharides production, designated as LZ-4T, was isolated from cultivable phycosphere microbiota of harmful algal blooms-causing marine dinoflagellate Alexandrium catenella LZT09 which produces paralytic shellfish poisoning toxins. Strain LZ-4T was able to use thiosulfate (optimum concentration 10 mM) as energy source for bacterial growth. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain LZ-4T belonged to the genus Limnobacter, showing high 16S rRNA gene sequences similarities with L. thiooxidans DSM 13612T (99.4%), L. humi NBRC 11650T (98.2%) and L. litoralis NBRC 105857T (97.2%), respectively. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between LZ-4T and L. thiooxidans DSM 13612T were 78.9 and 21.9%, respectively. Both values were far lower than the thresholds (95–96% for ANI and 70% for dDDH) generally accepted for new species delineation. The respiratory quinone of strain LZ-4T was Q-8. The dominant cellular fatty acids were determined as summed feature 3 (C16:1 ω6c/ω7c), summed feature 8 (C18:1 ω6c/ω7c) and C16:0. Polar lipids profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified aminolipids and three unidentified polar lipids. The genomic DNA G+C content of strain LZ-4T was 52.5 mol%. Based on polyphasic characterization, strain LZ-4T represents a novel species of the genus Limnobacter, for which the name Limnobacter alexandrii sp. nov. is proposed. The type strain is LZ-4T (=CCTCC AB 2019004T =KCTC 72281T).

MicroRNAs (miRNAs) are key in the post-transcriptional regulation of gene expression and thus characterization of miRNAs and investigation of the relative abundance and specificity of tissue expression are essential for understanding gene expression in the golden snub-nosed monkey (GSM, Rhinopithecus roxellanae). Here, we report the first dataset of GSM miRNAs where we identified 460 miRNAs in seven tissues, with 246 conserved known mature miRNAs and 214 novel mature miRNAs. We determined miRNA abundance and expression in the seven tissues using a Tissue Specificity Index score and found that most novel GSM miRNAs showed a highly tissue-specific expression pattern. In particular, 67 novel miRNAs and the miR-34 family were expressed in abundance only in the lung. Five known miRNAs were highly abundant in digestive organs such as the pancreas and liver, and four novel miRNAs were highly expressed in the heart and muscle. Annotation of target genes of GSM miRNAs indicated that target genes were enriched in many important pathways, such as the HIF-1 signaling pathway and xenobiotic biodegradation-related pathways. Collectively, these results emphasize that miRNAs play important roles in GSM diet and high-elevation adaptation regulation. In summary, this study provides essential information on GSM miRNAs and will benefit further investigations of the function and mechanism of miRNAs in controlling gene expression in the GSM.

A taxonomic study was carried out on a novel algae-associated bacterial strain Z3-1T, which was isolated from phycosphere microbiota of toxic marine dinoflagellate Alexandrium tamarense 880. Cells of strain Z3-1T were Gram-stain-negative, rod-shaped and strictly aerobic and were motile by means of flagella. Strain Z3-1T grew at 25–42 °C, pH 5.0–10.0 and 1.0–5.0 % (w/v) NaCl. Strain Z3-1T reduced nitrate to nitrite, but did not reduce nitrite to nitrogen gas. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Z3-1T belongs to the genus Nitratireductor showing the highest sequence similarity (97.0 %) to Nitratireductor basaltis JCM 14935T. The average nucleotide identity and digital DNA–DNA hybridization relatedness between strain Z3-1T and type strains of genus Nitratireductor with available genome sequences were in the ranges of 72.4–74.4 % and 22.7–23.3 %, respectively. The major fatty acids were summed in feature 8 (C18:1 ω7c and/or C18:1 ω6c), C19:0 ω8c cyclo, C18:1 ω7c 11-metyl and iso-C17:0. The major polar lipids were determined as diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, two unidentified phospholipids and four unidentified polar lipids. The genomic DNA G+C content calculated from genome sequence was 65.6 mol%. Based on genotypic, chemotaxonomic and phenotypic data obtained, strain Z3-1T represents a novel species of the genus Nitratireductor , for which the name Nitratireductor alexandrii sp. nov. is proposed with the type strain Z3-1T (=KCTC 62458T=CCTCC AB 2017227T).

A phytochemical study of 80% ethanol extract from the aerial parts of Euphorbia helioscopia led to the isolation of three new jatrophane diterpenoids, euphoheliphanes A-C (1–3). Their structures we...

Due to the vegetation destruction and soil desertification caused by excessive exploitation at Ganzhou ion-type rare earth mine in the mid-1980s, it is essential to carry out ecological remediation. The symbiotic mycorrhiza formed by the developed perennial ryegrass (Lolium perenne L.) roots infected with arbuscular mycorrhizal fungi (AMF) can significantly improve the growth and resistance of plants. In this study, the combination of symbiotic mycorrhiza and soil modifier was used to construct the ryegrass-AMF-soil modifier combined remediation technology, which achieved effective ecological remediation of soil tailings. The orthogonal experiment of soil modifier showed that the most efficient formula for ryegrass biomass, soil organic matter, soil alkaline hydrolysis, soil available phosphorus, and soil pH was 5 g/kg sepiolite, 3 g/kg chicken manure, 2 g/kg humic acid, and 2 g/kg biochar (A4B3C3D3), and chicken manure (B), humic acid (C), and biochar (D) had significant effects on the improvement of ryegrass biomass, soil organic matter, soil alkaline nitrogen, and soil available phosphorus. Sepiolite (A) had a significant improvement in soil pH. Furthermore, the AMF infection results indicated that Glomus moss (G.m.) had higher affinity with ryegrass. The T4 treatment-combined remediation using G.m. inoculation had the most significant effect on ryegrass growth; plant height increased by 39.19% compared with T1 treatment-inoculation using G.m. Under combined remediation, soil pH, organic matter, alkali nitrogen, and effective phosphorus content also significantly improved after combined treatment. Under G.m. inoculation treatment (T4 treatment), the soil nutrient content reached the three criteria of the soil nutrient grading standard.

A Gram-negative, aerobic, non-motile, non-spore-forming and rod-shaped bacterium, named strain LZ-16-2T, was isolated from the phycosphere microbiota of the paralytic shellfish poisoning toxin-producing marine dinoflagellate Alexandrium catenella LZT09. Strain LZ-16-2T grew optimally at 28 °C at pH 6.5 and with 3 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence revealed that strain LZ-16-2T fell within the genus Haliea and was most closely related to Haliea salexigens DSM 19537T, with which the new isolate exhibited 98.5 % 16S rRNA gene sequence similarity. The major respiratory quinone was Q-8. The predominant cellular fatty acids were C17 : 1 ω8c, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), C17 : 1 ω6c, C11 : 0 3-OH and C17 : 0. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The average nucleotide identity and in silico DNA–DNA genome hybridization relatedness values between strain LZ-16-2T and its closest relative, H. salexigens DSM 19537T, were 92.8 and 55.1 %, respectively. The DNA G+C content was 61.3 mol%. Differential phenotypic properties and phylogenetic distinctiveness distinguished strain LZ-16-2T from all other members of the genus Haliea . On the basis of the polyphasic characterization, strain LZ-16-2T represents a novel species of the genus Haliea , for which the name Haliea alexandrii sp. nov. is proposed. The type strain is LZ-16-2T (=KCTC 62344T=CCTCC AB2017229T).

A Gram-stain-negative, strictly aerobic, non-motile and non-pigmented spirillum, designated strain LZ-5T, was isolated from cultures of the paralytic shellfish poisoning (PSP) toxin-producing marine dinoflagellate Alexandrium catenella LZT09 collected from the Zhoushan sea area in the East China Sea during an algal bloom. The isolate grew at 4–40 °C (optimum, 25–33 °C) and pH 5.0–9.0 (optimum, 7.5) in the presence of 0.5–10 % (w/v) NaCl (optimum, 4.0 %). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain LZ-5T clearly belonged to the genus Saccharospirillum of the family Saccharospirillaceae . Strain LZ-5T shared highest 16S rRNA gene sequence similarity with Saccharospirillum impatiens EL-105T (98.9 %), Saccharospirillum mangrovi HK-33T (97.2 %), Saccharospirillum correiae CPA1T (96.8 %), Saccharospirillum salsuginis YIM-Y25T (96.8 %) and Saccharospirillum aestuarii IMCC 4453T (95.1 %). The average nucleotide identity and in silico DNA–DNA hybridization between strain LZ-5T and the two most closely related Saccharospirillum strains, S. impatiens EL-105T and S. mangrovi HK-33T, were 82.2 and 19.3 %, and 72.2 and 13.2 %, respectively. The predominant respiratory quinone of strain LZ-5T was Q-8, and the major fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and C16 : 0. The polar lipids of strain LZ-5T were diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), phosphatidylcholine (PC), glycolipid (GL), two unidentified glycophospholipids (GPLs), three unidentified aminophospholipids (APLs) and two unidentified lipids. The genomic DNA G+C content was 57.2 mol%. On the basis of this polyphasic characterization, strain LZ-5T represents a novel species of the genus Saccharospirillum , for which the name Saccharospirillum alexandrii sp. nov. is proposed. The type strain is LZ-5T (=KCTC 62460T=CCTCC AB2017232T).

A novel short-rod-shaped bacterial strain with poly-β-hydroxybutyric acid granules inside, designated as Z7-4T, was isolated from a culture of a marine dinoflagellate with palytoxin-producing capacity, Ostreopsis lenticularis OS06, collected from the East China Sea. Cells of Z7-4T were Gram-stain-negative, non-motile, strictly aerobic, 0.9–1.2 µm wide and 2.0–3.9 µm long. Growth occurred in 1–4 % (w/v) NaCl, at 15–37 °C and at pH 5.0–10.0, with optimum growth in 3.5 % (w/v) NaCl, at 30 °C and at pH 7.0. Analysis of 16S rRNA gene sequence revealed that Z7-4T shared the highest 16S rRNA gene sequence similarities with Nioella aestuarii JCM 30752T (98.8 %), followed by Nioella sediminis KCTC 42144T (98.6 %) and Nioella nitratireducens KCTC 32417T (96.9 %). Phylogenetic analysis based on nearly complete 16S rRNA gene sequences revealed that Z7-4T clearly represented a member of the genus Nioella within the family Rhodobacteraceae . The respiratory quinone of Z7-4T was identified as Q-10. Polar lipids of Z7-4T were phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, three unidentified aminophospholipids and one unidentified phospholipid. The major fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and C16 : 0. The DNA G+C content of Z7-4T was 63.0 mol%. DNA–DNA hybridization values of the isolate against the closely related type strains were far below the 70 % limit for species delineation. The average nucleotide identity and in silico DNA–DNA genome hybridization relatedness between Z7-4T and the closely related members of the genus Nioella , N. sediminis KCTC 42144T and N. nitratireducens KCTC 32417T, were 91.1 and 46.3 %, and 79.3 and 19.4 %, respectively. On the basis of the results of polyphasic analysis, Z7-4T is proposed to represent a novel species of the genus Nioella , for which the name Nioella ostreopsis sp. nov. is proposed. The type strain of Nioella ostreopsis is Z7-4T (=KCTC 62459T=CCTCC AB 2017231T)

Objectives: To compare the efficacy and safety of anterior temporal lobotomy (ATLo) and anterior temporal lobectomy (ATLe) in drug-resistant temporal lobe epilepsy.Methods: Patients diagnosed with pharmacoresistant temporal lobe epilepsy who underwent anterior temporal lobotomy (ATLo) or anterior temporal lobectomy (ATLe) performed by a single surgeon were retrospectively included. Every patient was followed up annually after surgery. The postoperative seizure outcome evaluation was based on the Engel and ILAE classifications. We compared postoperative complications and 2-year follow-up seizure outcomes between the ATLo group and the ATL group.Results: A total of 42 individuals (21 ATLo and 20 ATLe) were included. At the two-year follow-up, more patients in the ATLo group than the ATLe group had reached Engel class I (20 versus 14) and ILAE I (19 versus 13). However, these differences were not significant. One patient in the ATLo group had intraparenchymal hematoma and fully recovered. The two groups had similar incidences of other short-term complications, and no patients died or had any permanent complications.Discussion: ATLo is not inferior to ATLe for patients with drug-resistant temporal epilepsy. There was no significant difference in seizure outcomes or the rate of postoperative complications between the two groups. A large sample randomized control study is needed.

Homocysteine (Hcy) accelerates neuronal senescence and induces age-related neurodegenerative diseases. Silence signal regulating factor 1 (SIRT1) prolongs lifespan and takes neuroprotective effects. We have previously demonstrated that hydrogen sulfide (H2S) prevents Hcy-induced apoptosis of neuronal cells and has neuroprotective effect. In the present work, we aimed to investigate whether H2S protects HT22 cells against Hcy-induced neuronal senescence and whether SIRT1 mediates this role of H2S. We found that Hcy induced cellular senescence in HT22 cells, as determined by β-galactosidase staining, expressions of P16INK4a, P21CIPL, and trypan blue Staining, which are the markers of cellular senescence. However, sodium hydrosulfide (NaHS, the donor of H2S) significantly reversed Hcy-induced cellular senescence. Interestingly, NaHS not only up-regulated the expression of SIRT1 in HT22 cells but also reversed Hcy-downregulated the expression of SIRT1 in HT22 cells. Furthermore, we found that pretreatment with Sirtinol (an inhibitor of SIRT1) markedly reversed the protection of NaHS against Hcy-induced HT22 cells senescence and apoptosis. Our findings illustrated that H2S protects HT22 cells against Hcy-induced senescence by up-regulating SIRT1.

Background pUL21 is a conserved protein of Alphaherpesvirinae that performs multiple important functions. The C-terminus of pUL21 in other members of this subfamily has RNA-binding ability; this domain contributes to pseudorabies virus (PRV) retrograde axonal transport in vitro and in vivo and participates in newly replicated viral DNA packaging and intracellular virus transport. However, knowledge regarding duck enteritis virus (DEV) pUL21 is limited. Results We verified that DEV UL21 is a γ2 gene that encodes a structural protein. Moreover, we observed that pUL21 localized to the nucleus and cytoplasm. DEV pUL21 interacted with pUL16 and formed a complex in transfected human embryonic kidney (HEK) 293 T cells and DEV-infected duck embryo fibroblasts (DEFs). These results were further confirmed by CO-IP assays. Conclusions The DEV UL21 gene is a late gene, and pUL21 localizes to the nucleus and cytoplasm. DEV UL21 is a virion component. In addition, pUL21 can interact with pUL16. These findings provide insight into the characteristics of UL21 and the interaction between pUL21 and its binding partner pUL16. Our study enhances the understanding of DEV pUL21.

To investigate the pivotal roles of the duck plague virus (DPV) tegument protein UL14 in viral replication, we generated 2 mutated viruses of DPV by using the bacterial artifcial chromosome system, the UL14-null mutant virus (CHv-BAC-ΔUL14) and the corresponding revertant virus (CHv-BAC-ΔUL14R). We found that the CHv-BAC-ΔUL14 viruses exhibited impaired virion morphogenesis in transmission electron microscopy (TEM) studies. Furthermore, CHv-BAC-ΔUL14 exhibited a plaque size reduction in duck embryo fibroblasts (DEFs). Finally, CHv-BAC-ΔUL14 exhibited a significant viral growth defect. Taken together, our findings suggest that DPV UL14 protein regulates viral morphogenesis for efficient viral replication.

Zygosaccharomyces rouxii is an osmotolerant and halotolerant yeast that can participate in fermentation. To understand the mechanisms of salt and sugar tolerance, the transcription levels of Z. rouxii M 2013310 under 180 g/L NaCl stress and 600 g/L glucose stress were measured. The transcriptome analysis showed that 2227 differentially expressed genes (DEGs) were identified under 180 g/L NaCl stress, 1530 DEGs were identified under 600 g/L glucose stress, and 1278 DEGs were identified under both stress conditions. Then, KEGG enrichment analyses of these genes indicated that 53.3% of the upregulated genes were involved in the ergosterol synthesis pathway. Subsequently, quantitative PCR was used to verify the results, which showed that the genes of the ergosterol synthesis pathway were significantly upregulated under 180 g/L NaCl stress. Finally, further quantitative testing of ergosterol and spotting assays revealed that Z. rouxii M 2013310 increased the amount of ergosterol in response to high salt stress. These results highlighted the functional differences in ergosterol under sugar stress and salt stress, which contributes to our understanding of the tolerance mechanisms of salt and sugar in Z. rouxii.

Duck plague virus (DPV) pUL48 is a homologous of herpes simplex virus VP16, and some studies have shown that VP16 is essential for viral replication and proliferation, but there are few studies on DPV pUL48. Therefore, in order to study the function of pUL48 protein, we constructed a UL48-deleted mutant (DPV-BAC-∆UL48) that completely reemoved the UL48 gene from the DPV BAC genome and the revertant virus (DPV-BAC-∆UL48R) by using the 2-step red recombination system. Compared with the parental virus (DPV-BAC) and the revertant virus, the titer of UL48-deleted mutant was reduced by more than 38.2%, and the efficiency of producing infectious virions was significantly reduced. In addition, the average size of plaques produced by UL48-deleted mutant was about 30% smaller than that of the parental and revertant viruses, suggesting that pUL48 protein affected the cell-to-cell transmission of DPV. Finally, pharmacological inhibition assay showed that pUL48 is a late protein of DPV. In this study, we found that UL48, as a late gene, plays an important role in viral replication by affecting the formation of DPV infectious virion, virus cell-to-cell transmission, and viral genome transcription, which may provide some help for the study of the function of DPV pUL48 protein and the prevention and control of DPV.

Although the role of the flavivirus non-structural 2B (NS2B) as a viral protease cofactor has been investigated, its role in viral proliferation has rarely been studied. Moreover, important details on the involvement of the flavivirus NS2B in the virus replication cycle, including attachment, entry, RNA replication, assembly and release, remain unknown. Here, an alignment of flavivirus NS2B was performed to select conserved amino acids for the site-directed mutagenesis study. G92, which is not totally conserved to mutation, was also used. Certain mutations (P32A, G36A and G37A) completely blocked viral RNA synthesis, four mutations (D25A, A43Y, V44A, G92A and P112A) showed a slight defect in virus replication, and only the G92A mutant affected virus proliferation by blocking virus release but not RNA replication. By taking advantage of the tembusu virus (TMUV) replicon that represents viral genome RNA replication, we showed that the G92A mutation did not obviously impair viral replication. Naturally, the NS2B3 G92A mutation protein did not affect the self-cleavage ability or NS2B3 protein-mediated cleavage of the NS2AB G92A mutant. These results indicated that the G92A mutation did not affect viral protease activity. By using the TMUV infection clone and the replicon package system, the effect of G92 on the virus lifecycle was narrowed to viral resemblance or release. Subcellular fractionation further confirmed that G92A attenuated TMUV by reducing virus assembly. Collectively, these studies extend the list of NS2B functions beyond protease activity as a cofactor, show that transmembrane region amino acids play an important role in viral replication and indicate that G92 is involved in the assembly of infectious TMUV particles.