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Abstract Dracaena cambodiana Pierre ex Gagn. (Asparagaceae) is the source plant of Dragon’s blood and has high ornamental values in gardening. Currently, this species is classified as the second-class state-protected species in the National Key Protected Wild Plants (NKPWP) of China. However, limited genomic data has hindered a more comprehensive scientific understanding of the processes involved in the production of Dragon’s blood and the related conservation genomics research. In this study, we assembled a haplotype-resolved genome of D. cambodiana. The haploid genomes, haplotype A and haplotype B, are 1,015.22 Mb and 1,003.13 Mb in size, respectively. The completeness of haplotype A and haplotype B genomes was 98.60% and 98.20%, respectively, using the “embryophyta_10” dataset. Haplotype A and haplotype B genomes contained 27,361 and 27,066 protein-coding genes, respectively, with nearly all being functionally annotated. These findings provide new insights into the genomic characteristics of D. cambodiana and will offer additional genomic resources for studying the biosynthesis mechanism of Dragon’s blood and the horticultural application of Dragon trees.

IntroductionCodonopsis pilosula is a significant plant in traditional Chinese medicine, valued for its edible and medicinal properties. However, the lack of available genomic resources has hindered further research.MethodsThis study presents the first chromosome-scale genome assembly of C. pilosula using PacBio CLR reads and Hi-C scaffolding technology. Additionally, Ks analysis and syntenic depth analysis were performed to elucidate its evolutionary history.ResultsThe final assembly yielded a high-quality genome of 679.20 Mb, which was anchored to 8 pseudo-chromosomes with an anchoring rate of 96.5% and a scaffold N50 of 80.50 Mb. The genome assembly showed a high completeness of 97.6% based on Benchmarking with Universal Single-Copy Orthologs (BUSCO) analysis. Repetitive elements constituted approximately 76.8% of the genome, with long terminal repeat retrotransposons (LTRs) accounting for about 39.17%. Ks and syntenic depth analyses revealed that the polyploidization history of three platycodonoid clade species involved only the γ-WGT event. Karyotype evolutionary analysis identified an ancestral karyotype with 9 protochromosomes for the three platycodonoid clade species. Moreover, non-WGD genes, particularly those arising from tandem duplications, were found to contribute significantly to gene family expansion.DiscussionThese findings provide essential insights into the genetic diversity and evolutionary biology of C. pilosula, aiding its conservation and sustainable use.

Microplastics (MPs) and okadaic acid (OA) are known to coexist in marine organisms, potentially impacting humans through food chain. However, the combined toxicity of OA and MPs remains unknown. In this study, mice were orally administered OA at 200 μg/kg bw and MPs at 2 mg/kg bw. The co-exposure group showed a significant increase in malondialdehyde (MDA) content and significant decreases in superoxide dismutase (SOD) activity and glutathione (GSH) level compared to the control, MPs and OA groups (p < 0.05). Additionally, the co-exposure group exhibited significantly higher levels of IL-1β and IL-18 compared to other groups (p < 0.05). These results demonstrated that co-exposure to MPs and OA induces oxidative stress and exacerbates inflammation. Histological and cellular ultrastructure analyses suggested that this combined exposure may enhance gut damage and compromise barrier integrity. Consequently, the concentration of OA in the small intestine of the co-exposure group was significantly higher than that in the OA group. Furthermore, MPs were observed in the lamina propria of the gut in the co-exposure group. Transcriptomic analysis revealed that the co-exposure led to increased expression of certain genes related to the NF-κB/NLRP3 pathway compared to the OA and MPs groups. Overall, this combined exposure may disrupt the intestinal barrier, and promote inflammation through the NF-κB/NLRP3 pathway. These findings provide precious information for the understanding of health risks associated with MPs and phycotoxins.

Lamiales, comprising over 23,755 species across 24 families, stands as a highly diverse and prolific plant group, playing a significant role in the cultivation of horticultural, ornamental, and medicinal plant varieties. Whole-genome duplication (WGD) and its subsequent post-polyploid diploidization (PPD) process represent the most drastic type of karyotype evolution, injecting significant potential for promoting the diversity of this lineage. However, polyploidization histories, as well as genome and subgenome fractionation following WGD events in Lamiales species, are still not well investigated. In this study, we constructed a chromosome-level genome assembly of Lindenbergia philippensis (Orobanchaceae) and conducted comparative genomic analyses with 14 other Lamiales species. L. philippensis is positioned closest to the parasitic lineage within Orobanchaceae and has a conserved karyotype. Through a combination of Ks analysis and syntenic depth analysis, we reconstructed and validated polyploidization histories of Lamiales species. Our results indicated that Primulina huaijiensis underwent three rounds of diploidization events following the γ-WGT event, rather than two rounds as reported. Besides, we reconfirmed that most Lamiales species shared a common diploidization event (L-WGD). Subsequently, we constructed the Lamiales Ancestral Karyotype (LAK), comprising 11 proto-chromosomes, and elucidated its evolutionary trajectory, highlighting the highly flexible reshuffling of the Lamiales paleogenome. We identified biased fractionation of subgenomes following the L-WGD event across eight species, and highlighted the positive impacts of non-WGD genes on gene family expansion. This study provides novel genomic resources and insights into polyploidy and karyotype remodeling of Lamiales species, essential for advancing our understanding of species diversification and genome evolution.

Abstract Background Conus, a highly diverse species of venomous predators, has attracted significant attention in neuroscience and new drug development due to their rich collection of neuroactive peptides called conotoxins. Recent advancements in transcriptome, proteome, and genome analyses have facilitated the identification of conotoxins within Conus’ venom glands, providing insights into the genetic features and evolutionary patterns of conotoxin genes. However, the underlying mechanism behind the extraordinary hypervariability of conotoxins remains largely unknown. Results We analyzed the transcriptomes of 34 Conus species, examining various tissues such as the venom duct, venom bulb, and salivary gland, leading to the identification of conotoxin genes. Genetic variation analysis revealed that a subset of these genes (15.78% of the total) in Conus species underwent positive selection (Ka/Ks > 1, p

Abstract The treatment of chronic and non-healing wounds in diabetic patients remains a major medical problem. Recent reports have shown that hydrogel wound dressings might be an effective strategy for treating diabetic wounds due to their excellent hydrophilicity, good drug-loading ability and sustained drug release properties. As a typical example, hyaluronic acid dressing (Healoderm) has been demonstrated in clinical trials to improve wound-healing efficiency and healing rates for diabetic foot ulcers. However, the drug release and degradation behavior of clinically-used hydrogel wound dressings cannot be adjusted according to the wound microenvironment. Due to the intricacy of diabetic wounds, antibiotics and other medications are frequently combined with hydrogel dressings in clinical practice, although these medications are easily hindered by the hostile environment. In this case, scientists have created responsive-hydrogel dressings based on the microenvironment features of diabetic wounds (such as high glucose and low pH) or combined with external stimuli (such as light or magnetic field) to achieve controllable drug release, gel degradation, and microenvironment improvements in order to overcome these clinical issues. These responsive-hydrogel dressings are anticipated to play a significant role in diabetic therapeutic wound dressings. Here, we review recent advances on responsive-hydrogel dressings towards diabetic wound healing, with focus on hydrogel structure design, the principle of responsiveness, and the behavior of degradation. Last but not least, the advantages and limitations of these responsive-hydrogels in clinical applications will also be discussed. We hope that this review will contribute to furthering progress on hydrogels as an improved dressing for diabetic wound healing and practical clinical application.

Abstract Background Central pontine myelinolysis (CPM) is a rare demyelinating disorder caused by the loss of myelin in the center of the basis pontis. CPM typically occurs with rapid correction of severe chronic hyponatremia and subsequent disturbances in serum osmolality. Although hyperglycaemia is recognized as a pathogenetic factor in serum osmolality fluctuations, CPM is rarely seen in the context of diabetes. Case presentation A 66-year-old Chinese male presented with a history of gait imbalance, mild slurred speech and dysphagia for two weeks. MRI showed the mass lesions in the brainstem, and laboratory examinations showed high blood glucose and HbA1c, as well as increased serum osmolality. The patient was diagnosed with CPM secondary to hyperosmolar hyperglyceamia and received insulin treatment as well as supportive therapy. After six weeks of followup, the patient had fully recovered to a normal state. Conclusion CPM is a potentially fatal neurological condition and can occur in uncontrolled diabetes mellitus. Early diagnosis and timely treatment are crucial for improving the prognosis.

Triazine herbicides are common contaminants in coastal waters, and they are recognized as inhibitors of photosystem II, causing significant hinderance to the growth and reproduction of phytoplankton. However, the influence of these herbicides on microalgal toxin production remains unclear. This study aimed to examine this relationship by conducting a comprehensive physiological and 4D label-free quantitative proteomic analysis on the harmful dinoflagellate Karenia mikimotoi in the presence of the triazine herbicide dipropetryn. The findings demonstrated a significant decrease in photosynthetic activity and pigment content, as well as reduced levels of unsaturated fatty acids, reactive oxygen species (ROS), and hemolytic toxins in K. mikimotoi when exposed to dipropetryn. The proteomic analysis revealed a down-regulation in proteins associated with photosynthesis, ROS response, and energy metabolism, such as fatty acid biosynthesis, chlorophyll metabolism, and nitrogen metabolism. In contrast, an up-regulation of proteins related to energy-producing processes, such as fatty acid β-oxidation, glycolysis, and the tricarboxylic acid cycle, was observed. This study demonstrated that dipropetryn disrupts the photosynthetic systems of K. mikimotoi, resulting in a notable decrease in algal toxin production. These findings provide valuable insights into the underlying mechanisms of toxin production in toxigenic microalgae and explore the potential effect of herbicide pollution on harmful algal blooms in coastal environments.

Objectives: To evaluate the efficacy and safety of low-dose trimethoprim (TMP)-sulfamethoxazole (SMX) (TMP-SMX) as the primary prophylaxis for Pneumocystis jirovecii pneumonia (PJP) in adult recipients of kidney transplantation. Methods: Three kinds of prescriptions in kidney recipients were documented, including 20 mg TMP/100 mg SMX oral daily, 20 mg TMP/100 mg SMX oral every other day, and nonprophylaxis. The primary outcome was the incidence of PJP in the first 180 days of follow-up after kidney transplantation. The secondary outcomes were changes in renal and liver function. Results: Among the 1469 recipients, 1066 (72.56%) received 20 mg TMP/100 mg SMX daily, 127 (8.65%) received 20 mg TMP/100 mg SMX every other day, and 276 (18.79%) did not have prophylaxis prescription. The 276 recipients in the nonprophylaxis group had 124.92 person-years of follow-up, during which PJP occurred in 29 patients, for an incidence rate of 23.21 (95% confidence interval 15.76-32.72) per 100 person-years. The TMP-SMX daily group and the TMP-SMX every other day group had 524.89 and 62.07 person-years of follow-up, respectively, with no occurrence of PJP. There was no significant difference among the three groups in changes in renal and liver function (P >0.05, respectively). A total of 111 recipients in each group were enrolled in the propensity score matching analysis. It was revealed that the 111 nonprophylaxis recipients had 51.27 person-years of follow-up and 10 PJP cases. Prophylaxis was considered effective because there was a significant difference between the three groups (P

Okadaic acid (OA) is one of the most prevalent marine phycotoxin with complex toxicity, which can lead to toxic symptoms such as diarrhea, vomiting, nausea, abdominal pain, and gastrointestinal discomfort. Studies have shown that the main affected tissue of OA is digestive tract. However, its toxic mechanism is not yet fully understood. In this study, we investigated the changes that occurred in the epithelial microenvironment following OA exposure, including the epithelial barrier and gut bacteria. We found that impaired epithelial cell junctions, mucus layer destruction, cytoskeletal remodeling, and increased bacterial invasion occurred in colon of rats after OA exposure. At the same time, the gut bacteria decreased in the abundance of beneficial bacteria and increased in the abundance of pathogenic bacteria, and there was a significant negative correlation between the abundance of pathogenic bacteria represented by Escherichia/Shigella and animal body weight. Metagenomic analysis inferred that Escherichia coli and Shigella spp. in Escherichia/Shigella may be involved in the process of cytoskeletal remodeling and mucosal layer damage caused by OA. Although more evidence is needed, our results suggest that opportunistic pathogens may be involved in the complex toxicity of OA during OA-induced epithelial barrier damage.

Abstract Total joint arthroplasty is a common surgical procedure resulting in improved quality of life; however, a leading cause of surgery failure is infection. Periprosthetic joint infections often involve biofilms, making treatment challenging. The metabolic state of pathogens in the joint space and mechanism of their tolerance to antibiotics and host defenses are not well understood. Thus, there is a critical need for increased understanding of the physiological state of pathogens in the joint space for development of improved treatment strategies toward better patient outcomes. Here, we present a quantitative, untargeted NMR-based metabolomics strategy for Pseudomonas aeruginosa suspended culture and biofilm phenotypes grown in bovine synovial fluid as a model system. Significant differences in metabolic pathways were found between the suspended culture and biofilm phenotypes including creatine, glutathione, alanine, and choline metabolism and the tricarboxylic acid cycle. We also identified 21 unique metabolites with the presence of P. aeruginosa in synovial fluid and one uniquely present with the biofilm phenotype in synovial fluid. If translatable in vivo, these unique metabolite and pathway differences have the potential for further development to serve as targets for P. aeruginosa and biofilm control in synovial fluid.

Poleroviruses, which are distributed worldwide, infect many crops of economic importance and cause severe plant diseases. Brassica yellows virus (BrYV), which has three genotypes, A, B, and C, is a newly identified polerovirus infecting crucifer crops in China, but its distribution is still unclear. Here, we report the distribution and prevalence levels of the three BrYV genotypes in crucifer crops in China. A total of 570 crucifer leaf samples randomly collected from 22 provinces, four ethnic minority autonomous regions, and three municipalities in China were tested for BrYV. RT-PCR detection showed that 97 of the field samples were positive for BrYV, and the average incidence of BrYV was 17.0%. The virus was detected in 22 provinces, with high incidences in north, northwest, and northeast China. The multiplex RT-PCR amplification of the three BrYV genotypes revealed that both single and mixed infections occurred. Among the BrYV infections, 38.1% were mix-infected by more than two viral genotypes, and 8.2% samples were mix-infected by three viral genotypes. Our findings indicated a widespread prevalence of BrYV in China, and BrYV mixed infections with Turnip mosaic virus and Cucumber mosaic virus in crucifer crops are common. This study is the first large-scale survey of BrYV in crucifer crops in China. The information generated in this investigation will contribute to the national prevention and control of viral diseases.

BackgroundRenal ischemia-reperfusion injury (IRI) is an inevitable occurrence during kidney transplantation. Mitophagy, ferroptosis, and the associated immune microenvironment (IME) have been shown to play important roles in renal IRI. However, the role of mitophagy-associated IME genes in IRI remains unclear. In this study, we aimed to construct a prediction model of IRI prognosis based on mitophagy-associated IME genes.MethodThe specific biological characteristics of the mitophagy-associated IME gene signature were comprehensively analyzed using public databases such as GEO, Pathway Unification, and FerrDb. Correlations between the expression of prognostic genes and immune-related genes and IRI prognosis were determined by Cox regression, LASSO analysis, and Pearson’s correlation. Molecular validation was performed using human kidney 2 (HK2) cells and culture supernatant as well as the serum and kidney tissues of mice after renal IRI. Gene expression was measured by PCR, and inflammatory cell infiltration was examined by ELISA and mass cytometry. Renal tissue damage was characterized using renal tissue homogenate and tissue sections.ResultsThe expression of the mitophagy-associated IME gene signature was significantly correlated with IRI prognosis. Excessive mitophagy and extensive immune infiltration were the primary factors affecting IRI. In particular, FUNDC1, SQSTM1, UBB, UBC, KLF2, CDKN1A, and GDF15 were the key influencing factors. In addition, B cells, neutrophils, T cells, and M1 macrophages were the key immune cells present in the IME after IRI. A prediction model for IRI prognosis was constructed based on the key factors associated with the mitophagy IME. Validation experiments in cells and mice indicated that the prediction model was reliable and applicable.ConclusionWe clarified the relationship between the mitophagy-related IME and IRI. The IRI prognostic prediction model based on the mitophagy-associated IME gene signature provides novel insights on the prognosis and treatment of renal IRI.

Abstract Background There is a lack of data regarding the appropriateness of transperitoneal and retroperitoneal approaches for homolateral laparoscopic adrenalectomy. The aim of this study is to compare lateral transperitoneal and retroperitoneal approach for left-sided and right-sided laparoscopic adrenalectomy respectively. Methods Between January 2014 and December 2019, 242 patients underwent left-sided and 252 patients underwent right-sided laparoscopic adrenalectomy. For left side, transperitoneal approach was used in 132 (103 with tumors

Gambierdiscus spp. is mainly responsible for the ciguatera fish poisoning (CFP) around the world. The gambiertoxin produced by Gambierdiscus can be passed through the food chain to form ciguatoxins (CTXs) that cause ciguatoxins poisoning. However, the toxic effects of Gambierdiscus on fish through the food chain and related mechanism remains unclear. In this study, the toxicity of Gambierdiscus caribaeus on the marine medaka (Oryzias melastigma) was investigated, where the simulated food chain toxic algae-food organism-fish (G. caribaeus-Artemia metanauplii-O. melastigma) was set. The results showed that direct or indirect exposure through the food chain of G. caribaeus could affect the swimming behaviour of O. melastigma, manifested as decreased swimming performance and spontaneous abnormal swimming behaviours. Histological observation showed that direct or indirect exposure of G. caribaeus caused different degrees of pathological damage to the gills, intestine and liver tissues of O. melastigma. Transcriptome sequencing and RT-qPCR demonstrated that G. caribaeus exposure could trigger a series of physiological and biochemical responses, mainly reflected in energy metabolism, reproductive system, neural activity, immune stress and drug metabolism in marine medaka. Our finding may provide novel insight into the toxicity of Gambierdiscus on fish.

The analysis of NMR spectra of complex biochemical samples with respect to individual resonances is challenging but critically important. Here, the authors present a deep learning-based method that accelerates this process also for crowded NMR data that are non-trivial to analyze, even by expert NMR spectroscopists.

Sedimentary nitrogen isotope (δ15Nsed) in Santa Barbara Basin (SBB) has been mostly interpreted as the record of the eastern tropical North Pacific (ETNP) intermediate water denitrification process. Nevertheless, debate remains regarding sources and control mechanisms of δ15Nsed signal in SBB. Multi-proxy analyses including δ15Nsed, total organic carbon (TOC), total nitrogen (TN), C/N ratio, and marine biomarkers were performed on a 46-cm sediment core (SBB-190629) collected from SBB in 2019. The core was dated with varve counting and 210Pb dating method, showing a depositional history of 1938–2019 CE with a sedimentation rate of 0.564 cm/year. The findings show that the δ15Nsed record (at ~0.25-year resolution) ranges from 6.24‰ to 7.43‰, which was affected by both local and remote forcing. The long-term variations of the SBB δ15Nsed signature show a general decreasing trend from 1940 to the late 1980s, low values during 1980~2000, and an increase afterward, which is thought to reflect changes in ETNP denitrification induced by the strength of tropical trade winds. Our results also reveal a series of abrupt annual to multiannual changes, superimposed on the long-term variation mentioned above. The SBB local δ15N signal (Δδ15NSBB) is accessed by using the deviation from the mean δ15N (Δδ15N) of SBB-190629 to subtract the Δδ15N of the ETNP. The Δδ15NSBB record compares well with redox-sensitive proxies (Re/Mo ratio and C29 stanol/stenol ratio) from the SBB bottom water and with the OCmarine content calculated based on the C/N ratio mixing model, indicating that the Δδ15NSBB is mainly controlled by bottom water denitrification, which was induced by the change of upwelling intensity and marine productivity. Since various climatic factors (e.g., El Niño–Southern Oscillation (ENSO), Pacific Decadal Oscillation (PDO), and North Pacific Gyre Oscillation (NPGO)) have different impacts on the upwelling intensity (hence the marine productivity and denitrification) in SBB on different timescales, the influence of combined climatic factors on SBB denitrification is time dependent.

ObjectiveTo investigate the risk factors of infectious diseases in adult kidney transplantation recipients and to establish a simple and novel nomogram to guide the prophylactic antimicrobial therapy.MethodsPatients who received kidney transplantation between January 2018 and October 2021 were included in the study and were divided into a training and a testing set at a 1:1 ratio. Risk factors correlated to infectious diseases were selected using a Least Absolute Shrinkage and Selection Operator (LASSO) regression model. The prediction model was built by incorporating the variables selected by the LASSO model into a logistic regression equation. Calibration curves and receiver operating characteristic (ROC) curves were also applied to assess the model calibration and discrimination. A nomogram consisting of the selected factors was established to provide individualized risks of developing infections. Decision curve analysis (DCA) was adopted to estimate the net benefit and reduction in interventions for a range of clinically reasonable risk thresholds.ResultsIn all, 863 adult kidney recipients were included in the study, and 407 (47.16%) of them developed infectious diseases during the 3-year follow–up period. A total of 8 variables were selected using LASSO regression and were retained for subsequent model construction and infection prediction. The area under the curve (AUC) was 0.83 and 0.81 in the training and testing sets, with high F scores of 0.76 and 0.77, sensitivity of 0.76 and 0.81, and specificity of 0.88 and 0.74, respectively. A novel nomogram was developed based on 8 selected predictors (requirement for albumin infusion, requirement for red blood cell infusion, triglyceride, uric acid, creatinine, globulin, neutrophil percentage, and white blood cells). The net benefit indicated that the nomogram would reduce unnecessary interventions at a wide range of threshold probabilities in both sets.ConclusionsAdult kidney transplantation recipients are high-risk hosts for infectious diseases. The novel nomogram consisting of 8 factors reveals good predictive performance and may promote the reasonable antimicrobial prescription. More external validations are required to confirm its effectiveness for further clinical application.

Zearalenone (ZEN) is a potent oestrogenic mycotoxin that is mainly produced by Fusarium species and is a serious environmental pollutant in animal feeds. Apiotrichum mycotoxinivorans has been widely used as a feed additive to detoxify ZEN. However, the effects of ZEN on A. mycotoxinivorans and its detoxification mechanisms remain unclear. In this study, transcriptomic and bioinformatic analyses were used to investigate the molecular responses of A. mycotoxinivorans to ZEN exposure and the genetic basis of ZEN detoxification. We detected 1424 significantly differentially expressed genes (DEGs), of which 446 were upregulated and 978 were downregulated. Functional and enrichment analyses showed that ZEN-induced genes were significantly associated with xenobiotic metabolism, oxidative stress response, and active transport systems. However, ZEN-inhibited genes were mainly related to cell division, cell cycle, and fungal development. Subsequently, bioinformatic analysis identified candidate ZEN-detoxification enzymes. The Baeyer–Villiger monooxygenases and carboxylesterases, which are responsible for the formation and subsequent hydrolysis of a new ZEN lactone, respectively, were significantly upregulated. In addition, the expression levels of genes related to conjugation and transport involved in the xenobiotic detoxification pathway were significantly upregulated. Moreover, the expression levels of genes encoding enzymatic antioxidants and those related to growth and apoptosis were significantly upregulated and downregulated, respectively, which made it possible for A. mycotoxinivorans to survive in a highly toxic environment and efficiently detoxify ZEN. This is the first systematic report of ZEN tolerance and detoxification in A. mycotoxinivorans. We identified the metabolic enzymes that were potentially involved in detoxifying ZEN in the GMU1709 strain and found that ZEN-induced transcriptional regulation of genes is key to withstanding highly toxic environments. Hence, our results provide valuable information for developing enzymatic detoxification systems or engineering this detoxification pathway in other species.

Constructing optical nanoprobes with superior performance is highly desirable for sensitive and accurate assays. Herein, we develop a facile room-temperature strategy for the fabrication of green emissive carbon nanoclusters (CNCs) with dual-exciting centers for the dual-channel sensing of hemin. The formation of the CNCs is attributed to the crosslinking polymerization of the precursors driven by the Schiff base reaction between ethylenediamine and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone. Most importantly, the proposed CNCs have a unique excitation-independent green emission (518 nm) with two excitation centers at 260 nm (channel 1) and 410 nm (channel 2). The dual-exciting central emission can serve as dual-channel fluorescence (FL) signals for highly sensitive and reliable detection of hemin based on the inner filter effect. Because of the great spectral overlap difference between the absorption spectrum of hemin and the excitation lights of the CNCs in the two channels, hemin has a different quenching effect on FL emission from different channels. The dual-channel signals of the CNCs can detect hemin in the range of 0.075–10 μM (channel 1) and 0.25–10 μM (channel 2), respectively. These findings not only offer new guidance for the facile synthesis of dual-exciting central CNCs but also establish a reliable sensing platform for the analysis of hemin in complex matrixes.

Archaeal ammonia oxidation is the most important intermediate pathway in regulating the oceanic nitrogen cycle; however, the study of its specific role on a geological time scale is restricted to a specific part of marginal seas; thus far, only in the southern South China Sea (SCS). To explore the spatial pattern of the role of archaeal ammonia oxidation in the SCS, the GDGT-[2]/[3] ratio (Glycerol Dialkyl Glycerol Tetraether), an indicator of the archaeal ammonia oxidation rate, was analyzed and examined from the collected data profiles since the last glacial period in the northern SCS. The results showed that the GDGT-[2]/[3] ratio in the northern SCS was opposite to that in the southern SCS, with higher GDGT-[2]/[3] values during the Holocene compared to the last glacial period. Based on existing published depths of thermocline (DOT) data in the northern SCS since 30 ka, we believe that hydrological structural variations induced by mesoscale eddies caused this difference. Therefore, physical processes are very important factors that control the nitrogen cycle over a long-time scale. This study may provide new insights into the understanding of the role of archaeal ammonia oxidation within the marine nitrogen cycle over geological time scale.

Abstract To explore the characteristics and prognostic significance of genetic mutations in acute myeloid leukemia (AML), we screened the gene mutation profile of 171 previously untreated AML patients using a next‐generation sequencing technique targeting 127 genes with potential prognostic significance. A total of 390 genetic alterations were identified in 149 patients with a frequency of 87.1%. Younger age and high sensitivity to induction chemotherapy were associated with a lower number of mutations. NPM1 mutation was closely related to DNMT3A and FLT3‐internal tandem duplication (FLT3‐ITD) mutations, but mutually exclusive with ASXL1 mutation and CEBPAdouble mutation. In univariate analysis, ASXL1 or TET2 mutation predicted shorter overall survival (OS) or relapse‐free survival (RFS), DNMT3A, FLT3‐ITD, or RUNX1 mutation predicted a higher likelihood of remission‐induction failure, whereas NRAS mutation or CEBPAdouble mutation predicted longer OS. Concurrent DNMT3A, FLT3‐ITD, and NPM1 mutations predicted shorter OS. Hypomethylation agents could improve the OS in patients with DNA methylation‐related mutations. According to multivariate analysis, TET2 mutation was recognized as an independent prognostic factors for RFS. In summary, our study provided a detailed pattern of gene mutations and their prognostic relevance in Chinese AML patients based on targeted next‐generation sequencing screening.

Objective To investigate the polymorphism of T cell receptor β chain variable region (TCRVβ) in peripheral blood of clinically isoiated syndrome (CIS) patients. To preliminarily predict the specific TCRVβ fragment of CIS and its diagnostic value for CIS. Methods Hospitalized patients of Beijing Xuanwu Hospital from September 2012 to March 2014, were divided into 2 groups including CIS group (experimental pateints, n=30) and normal control group (healthy volunteers, n=30). The routine immune indexes and expression of TCRVβ subfamilies were detected by flow cytometry. CELL Quest Pro 5.1 software was used to analyze 24 subfamilies of TCRVβ. Calculated the statistic of amplification rate, amplification number and amplification fraction of the total 24 subfamilies of TCRVβ. Results Compared to the normal control group, the quantitative expression of CD4+TCRVβ2, 4, 5.2, 5.3, 7.1, 8, 14, 17, 20, 21.3 and CD8+TCRVβ4, 14 were significantly expanded in CIS group, CD4+TCRVβ11 and CD8+TCRVβ9, 11 were significantly reduced in CIS group (P

Abstract Cucurbit aphid-borne yellows virus (CABYV), melon aphid-borne yellows virus (MABYV) and suakwa aphid-borne yellows virus (SABYV) are three poleroviruses that infect cucurbit crops. Developing specific antisera against such viruses is crucial for their detection and functional understanding of related genes. However, no studies have yet reported viral detection using antisera against movement proteins (MP) in these three viruses. In this study, we generated plasmids expressing three viral MP genes, and transformed them into the Escherichia coli strain, Rosetta, to recombinantly express and purify fusion proteins. Then, polyclonal antisera were derived by immunizing New Zealand white rabbits, after which western blotting was used to determine the titer, sensitivity and specificity of the antisera. The antisera titers against MPCABYV, MPMABYV and MPSABYV were 1:512000, 1:256000 and 1:256000, respectively. The optimized working concentrations for the three antisera ranged between 1:10000 and 1:64000. Additionally, antisera against MPCABYV and MPMABYV only reacted with their corresponding MP proteins. Antiserum against MPSABYV not only had the strongest reaction with its MP, but also reacted weakly with MPCABYV and MPMABYV. All three antisera exerted no serological reactions with other poleroviruses. Furthermore, our data showed that all antisera specifically detected MPs in both Nicotiana benthamiana and cucumber leaves. Thus, we have established a system that sensitively detects three poleroviruses infecting cucurbits, using antisera against MPs. We provide a foundation for future research on the serological detection of these viruses, and interaction mechanisms between viruses and host plants.

The Huanghe (Yellow River) supplies large amount of sediments and terrestrial organic carbon (OC) to the eastern Chinese marginal seas. A relocation of the Huanghe outlet from the southern Yellow Sea (YS) to the Bohai Sea occurred in 1855 AD, however, detailed knowledge about the impact of this relocation on sedimentary source and OC burial in Chinese marginal seas is still critically lacking. In this study, we present total OC content and its isotope (δ13C), along with bulk total organic carbon (TOC)/total nitrogen (TN) molar ratio and lipid biomarker contents, in a sediment core HH12 from the southern YS with sediment age spanning the last 300 years. We find that TOC and terrestrial lipid biomarker mass accumulation rates were lower between 1855 AD and 1950 AD than that prior to 1855 AD in core HH12; and in accordance, both TOC/TN ratio and δ13C records indicate a gradual decrease of terrigenous source contributions to sedimentary OC. This suggests that the relocation of the Huanghe outlet reduced the transport of terrestrial OC to the southern YS. However, the δ13C record also indicates a relative increase of terrestrial OC contribution to sedimentary OC after 1950 AD, and the most likely explanation is increased contributions from the old Huanghe delta erosion and Korean rivers. Future studies should focus on better constraining the variations of terrestrial and marine endmembers with δ13C and Δ14C analyses of specific biomarkers to examine these linkages.

There is a critical need to accurately diagnose, prevent, and treat biofilms in humans. The biofilm forming P. aeruginosa bacteria can cause acute and chronic infections, which are difficult to treat due to their ability to evade host defenses along with an inherent antibiotic-tolerance. Using an untargeted NMR-based metabolomics approach, we identified statistically significant differences in 52 metabolites between P. aeruginosa grown in the planktonic and lawn biofilm states. Among them, the metabolites of the cadaverine branch of the lysine degradation pathway were systematically decreased in biofilm. Exogenous supplementation of cadaverine caused significantly increased planktonic growth, decreased biofilm accumulation by 49% and led to altered biofilm morphology, converting to a pellicle biofilm at the air-liquid interface. Our findings show how metabolic pathway differences directly affect the growth mode in P. aeruginosa and could support interventional strategies to control biofilm formation.

p53 is a vital transcriptional protein implicated in regulating diverse cellular processes, including cell cycle arrest, DNA repair, mitochondrial metabolism, redox homeostasis, autophagy, senescence, and apoptosis. Recent studies have revealed that p53 levels and activity are substantially increased in affected neurons in cellular and animal models of Parkinson’s disease (PD) as well as in the brains of PD patients. p53 activation in response to neurodegenerative stress is closely associated with the degeneration of dopaminergic neurons accompanied by mitochondrial dysfunction, reactive oxygen species (ROS) production, abnormal protein aggregation, and impairment of autophagy, and these pathogenic events have been implicated in the pathogenesis of PD. Pathogenic p53 integrates diverse cellular stresses and activate these downstream events to induce the degeneration of dopaminergic neurons; thus, it plays a crucial role in the pathogenesis of PD and appears to be a potential target for the treatment of the disease. We reviewed the current knowledge concerning p53-dependent neurodegeneration to better understand the underlying mechanisms and provide possible strategies for PD treatment by targeting p53.

Antibodies are one of the most important groups of biomolecules for both clinical and basic research and have been developed as potential therapeutics. Affinity is the key feature for biological activity and clinical efficacy of an antibody, especially of therapeutic antibodies, and thus antibody affinity improvement is indispensable and still remains challenging. To address this issue, we developed the E. coliAssisted Speed affINity-maturation Evolution SyStem (EASINESS) for continuous directed evolution of Ag–Ab interactions. Two key components of EASINESS include a mutation system modified from error-prone DNA polymerase I (Pol I) that selectively mutates ColE1 plasmids in E. coli and a protein–protein interaction selection system from mDHFR split fragments. We designed a GCN4 variant which barely forms a homodimer, and during a single round of evolution, we reversed the homodimer formation activity from the GCN4 variant to verify the feasibility of EASINESS. We then selected a potential therapeutic antibody 18A4Hu and improved the affinity of the antibody (18A4Hu) to its target (ARG2) 12-fold in 7 days while requiring very limited hands-on time. Remarkably, these variants of 18A4Hu revealed a significant improved ability to inhibit melanoma pulmonary metastasis in a mouse model. These results indicate EASINESS could be as an attractive choice for antibody affinity maturation.

Objective To explore effect of internal carotid artery (ICA) stenting on mild cognitive impairment (MCI) of patients with asymptomatic ICA stenosis. Methods Fifty-four patients with severe ICA stenosis (stenosis rate ≥ 70%) from October 2016 to September 2018 were studied. Cognitive function was examined in 54 patients with asymptomatic ICA stenosis 24 h before operation and 1 month, 3 months, 6 months after operation. Cognitive function was assessed with the Mini-Mental State Examination (MMSE), Trail Making Test (TMT) and Stroop Color-Word Test (SCWT). Results There were statistical differences in MMSE (P = 0.000), time of TMT-A and TMT-B (P = 0.000, 0.000), time of SCWT-A, SCWT-B and SCWT-C (P = 0.000, 0.036, 0.000), and correct number of SCWT-A (P = 0.005) before and after ICA stenting. MMSE, time of TMT-A, TMT-B and SCWT-A at 3 and 6 months after stenting were shorter than before and one month after stenting (P < 0.05, for all), while time of SCWT-C was shorter than before stenting (P < 0.05, for all). Time of SCWT-B at 6 months after stenting was shorter than before stenting (P < 0.05, for all). Correct number of SCWT-A at 6 months after stenting was higher than before and 3 months after stenting (P < 0.05, for all), which at 3 monthes after stenting was lower than one month after stenting (P < 0.05, for all). Conclusions ICA stenting can help to improve the MCI patients with asymptomatic ICA stenosis. DOI:10.3969/j.issn.1672-6731.2019.10.007

Abstract The serological method is one of the most important techniques extensively used in crop production to detect different pathogens, especially plant viruses. An antiserum is essential for serological tests. The 17 kDa movement protein (MP) of Potato leafroll virus (PLRV) is related to the membranous structures and localized to the plasmodesmata, but there is no report on preparation of PLRV-MP antiserum for detection of PLRV. To prepare PLRV-MP antiserum, reverse transcription polymerase chain reaction (RT-PCR) was carried out to amplify the PLRV-MP gene, which was constructed into a prokaryotic vector to express the protein in Escherichia coli for immunization of rabbits, after purification. Western blotting revealed that this developed antiserum could effectively detect PLRV, but with better results from the perspective of color development and economics by using antiserum at the ratio range of 1:10000 to 1:40000, presenting high sensitivity and specificity to PLRV. The serological detection results for PLRV of the field samples were identical to the RT-PCR detection. This is the first report on the development of PLRV-MP antiserum that has been successfully used for both laboratory and field detection of PLRV. The results provide a fundamental tool for further research on the function of PLRV-MP.

Prorocentrum lima is a global benthic dinoflagellate that produces diarrhetic shellfish poisoning (DSP) toxins, which can be ingested by filter-feeding bivalves, and eventually pose a great threat to human health through food chain. After being exposed to P. lima, different bivalves may accumulate various levels of DSP toxins and display different toxic responses. However, the underlying mechanism remains unclear. Here, we found that the content of okadaic acid-equivalents (OA-eq) varied in the digestive glands of the three bivalves including Crassostrea gigas, Mytilus coruscus and Tegillarca granosa after P. lima exposure. The degree of esterification of OA-eq in the three bivalves were opposite to the accumulation of OA-eq. The digestive gland tissues of the three bivalve species were damaged to different degrees. The transcriptional induction of Nrf2 targeted genes such as ABCB1 and GPx indicates the functionality of Nrf2 pathway against DSP toxins in bivalves. The oyster could protect against DSP toxins mainly through ABC transporters and esterification, while the mussel and clam reduce the damage induced by DSP toxins mainly by regulating the expression of antioxidant genes. Our findings may provide some explanations for the difference in toxic response to DSP toxins in different shellfish.

Objective: To analyze the clinical features of children with psychogenic non-epileptic seizures in one tertiary center in China.Methods: Clinical data including medical records and video- electroencephalograph (video-EEG) monitoring records of 88 pediatric PNES patients hospitalized in the pediatric department of Xuanwu Hospital, Beijing, China from April, 2012 to April, 2018 were collected in this study. Demographic information of patients, semiological classification, duration, and frequency of symptoms, risk factors as well as comorbidity were summarized and analyzed.Results: For semiological classification, all PNES related symptoms were divided into different categories: motor symptoms, unresponsiveness, sensory symptoms, visceral symptoms, and abnormal behaviors, among which motor symptoms were the most prevalent form. Risk factors were reviewed and categorized into two groups: persistent factors and predisposing factors, and patients were most frequently affected by the influences of families. The duration and frequency of symptoms varied substantially within PNES patients while the average time of duration was relatively longer than epilepsy as reported previously. Epilepsy was considered as the most frequent comorbidity of PNES and PNES patients misdiagnosed as epilepsy often mistreated with antiseizure medication.Significance: Our study showed that motor PNES are the most frequent seizure type. Family issues were a risk factor for PNES. Epilepsy was the most frequent co-existing neurological comorbidity.

Accumulating studies suggest that neuroinflammation characterized by microglial overactivation plays a pivotal role in the pathogenesis of Parkinson’s disease. As such, inhibition of microglial overactivation might be a promising treatment strategy to delay the onset or slow the progression of Parkinson’s disease. Ginsenoside Rb1, the most active ingredient of ginseng, reportedly exerts neuroprotective effects by suppressing inflammation in vitro. The present study aimed to evaluate the neuroprotective and anti-inflammatory effects of ginsenoside Rb1 in a lipopolysaccharide-induced rat Parkinson’s disease model. Rats were divided into four groups. In the control group, sham-operated rats were intraperitoneally administered normal saline for 14 consecutive days. In the ginsenoside Rb1 group, ginsenoside Rb1 (20 mg/kg) was intraperitoneally injected for 14 consecutive days after sham surgery. In the lipopolysaccharide group, a single dose of lipopolysaccharide was unilaterally microinjected into the rat substantial nigra to establish the Parkinson’s disease model. Lipopolysaccharide-injected rats were treated with normal saline for 14 consecutive days. In the ginsenoside Rb1 + lipopolysaccharide group, lipopolysaccharide was unilaterally microinjected into the rat substantial nigra. Subsequently, ginsenoside Rb1 was intraperitoneally injected for 14 consecutive days. To investigate the therapeutic effects of ginsenoside Rb1, behavioral tests were performed on day 15 after lipopolysaccharide injection. We found that ginsenoside Rb1 treatment remarkably reduced apomorphine-induced rotations in lipopolysaccharide-treated rats compared with the lipopolysaccharide group. To investigate the neurotoxicity of lipopolysaccharide and potential protective effect of ginsenoside Rb1, contents of dopamine and its metabolites in the striatum were measured by high-performance liquid chromatography. Compared with the lipopolysaccharide group, ginsenoside Rb1 obviously attenuated the lipopolysaccharide-induced depletion of dopamine and its metabolites in the striatum. To further explore the neuroprotective effect of ginsenoside Rb1 against lipopolysaccharide-induced neurotoxicity, immunohistochemistry and western blot assay of tyrosine hydroxylase were performed to evaluate dopaminergic neuron degeneration in the substantial nigra par compacta. The results showed that lipopolysaccharide injection caused a large loss of tyrosine hydroxylase-immunoreactive neurons in the substantia nigra and a significant decrease in overall tyrosine hydroxylase expression. However, ginsenoside Rb1 noticeably reversed these changes. To investigate whether the neuroprotective effect of ginsenoside Rb1 was associated with inhibition of lipopolysaccharide-induced microglial activation, we examined expression of the microglia marker Iba-1. Our results confirmed that lipopolysaccharide injection induced a significant increase in Iba-1 expression in the substantia nigra; however, ginsenoside Rb1 effectively suppressed lipopolysaccharide-induced microglial overactivation. To elucidate the inhibitory mechanism of ginsenoside Rb1, we examined expression levels of inflammatory mediators (tumor necrosis factor-α, interleukin-1β, inducible nitric oxide synthase, and cyclooxygenase 2) and phosphorylation of nuclear factor kappa B signaling-related proteins (IκB, IKK) in the substantia nigra with enzyme-linked immunosorbent and western blot assays. Our results revealed that compared with the control group, phosphorylation and expression of inflammatory mediators IκB and IKK in the substantia nigra of lipopolysaccharide group rats were significantly increased; whereas, ginsenoside Rb1 obviously reduced lipopolysaccharide-induced changes on the lesioned side of the substantial nigra par compacta. These findings confirm that ginsenoside Rb1 can inhibit inflammation induced by lipopolysaccharide injection into the substantia nigra and protect dopaminergic neurons, which may be related to its inhibition of the nuclear factor kappa B signaling pathway. This study was approved by the Experimental Animal Ethics Committee of Shandong University of China in April 2016 (approval No. KYLL-2016-0148).

Though microalgae have been considered the potential resource for lipid production, native strains are unable to meet the industrial demand. Here, we aim to uncover the complex molecular relationship between algal growth and lipid accumulation. Transcriptomic analysis revealed the crucial role of plastidial fatty acid and triacylglycerol (TAG) biosynthetic machinery in lipid overproduction. The expression of key fatty acid biosynthetic genes such as acetyl-CoA carboxylase (ACCase), malonyl CoA−acyl carrier protein transacylase (MCAT), 3-ketoacyl synthase (KAS), 3-ketoacyl-ACP reductase (KAR) increased during day 10–13 of cultivation, particularly plastidial TAG biosynthetic genes substantially increased. However, expression of genes involved in ER TAG biosynthesis increased only in the stationary phase, which implied the potential of plastidial TAG biosynthesis. This report provides a novel insight into the growth-phase dependent lipogenic orchestration, and also uncovers the signature genes and plastidial TAG biosynthesis that might be extrapolated for improving lipogenic traits of algae.

Abstract Background Microalgal metabolic engineering holds great promise for the overproduction of a wide range of commercial bioproducts. It demands simultaneous manipulation of multiple metabolic nodes. However, high-efficiency promoters have been lacking. Results Here we report a strong constitutive promoter Pt211 in expressing multiple target genes in oleaginous microalga Phaeodactylum tricornutum. Pt211 was revealed to contain significant cis-acting elements. GUS reporter and principal genes glycerol-3-phosphate acyltransferase (GPAT) and diacylglycerol acyltransferase 2 (DGAT2) involved in triacylglycerol biosynthesis were tested under driven of Pt211 in P. tricornutum. GUS staining and qPCR analysis showed strong GUS expression. DGAT2 and GPAT linked with a designed 2A sequence exhibited higher transcript abundances than WT, while algal growth and photosynthesis were not impaired. Conclusion The total lipid content increased notably by 2.6-fold compared to WT and reached up to 57.5% (dry cell weight). Overall, our findings report a strong promoter and a strategy for coordinated manipulation of complex metabolic pathways.

Brassica yellows virus (BrYV) is a tentative species of the genus Polerovirus, which has at least three genotypes (A, B, and C) in China. The P0 protein of BrYV-A (P0BrA) has been identified as a viral suppressor of RNA silencing (VSR), which can also induce cell death in infiltrated Nicotiana benthamiana leaves. In this study, we demonstrated that the cell death induced by P0BrA was accompanied by the accumulation of reactive oxygen species (ROS) and increased Pathogenesis-related protein genes-1 (PR1) expression. Meanwhile, this cell death phenotype was delayed by salicylic acid (SA) pretreatment. Biological function comparison of the three P0 proteins showed that transiently expressed P0BrB or P0BrC induced a significantly delayed and milder cell death response compared with P0BrA. However, like P0BrA, they also suppressed local and systemic RNA silencing. Six residues of P0BrA essential for inducing cell death were identified by comparative analysis and amino acid substitution assay. We also show that all three BrYV genotypes have synergistic interactions with pea enation mosaic virus 2 (PEMV 2) in N. benthamiana. This study provides theoretical guidance for controlling the viral disease caused by poleroviruses in the future.

Brassica yellows virus (BrYV) is a tentative species of the genus Polerovirus, which occurs widely, and mostly damages Brassicaceae plants in East Asia. Because BrYV cannot be transmitted mechanically, an insect-based transmission method is required for further virus research. Here, a reliable and unrestricted method is described, in which non-viruliferous aphids (Myzus persicae) acquired BrYV from transgenic Arabidopsis thaliana, harboring the full-length viral genome germinated from seeds and its frozen leaves. The aphids then transmitted the virus to healthy plants. There was no significant difference in acquisition rates between fresh and frozen infected leaves, although the transmission rate from frozen infected leaves was lower compared to fresh infected leaves. This simple novel method may be used to preserve viral inocula, evaluate host varietal resistance to BrYV, and investigate interactions among BrYV, aphids, and hosts.

Abstract Background Microalgae have emerged as a potential feedstock for biofuels and bioactive components. However, lack of microalgal strains with promising triacylglycerol (TAG) content and desirable fatty acid composition have hindered its commercial feasibility. Attempts on lipid overproduction by metabolic engineering remain largely challenging in microalgae. Results In this study, a microalgal 1-acyl-sn-glycerol-3-phosphate acyltransferase designated AGPAT1 was identified in the model diatom Phaeodactylum tricornutum. AGPAT1 contained four conserved acyltransferase motifs I–IV. Subcellular localization prediction and thereafter immuno-electron microscopy revealed the localization of AGPAT1 to plastid membranes. AGPAT1 overexpression significantly altered the primary metabolism, with increased total lipid content but decreased content of total carbohydrates and soluble proteins. Intriguingly, AGPAT1 overexpression coordinated the expression of other key genes such as DGAT2 and GPAT involved in TAG synthesis, and consequently increased TAG content by 1.81-fold with a significant increase in polyunsaturated fatty acids, particularly EPA and DHA. Moreover, besides increased lipid droplets in the cytosol, ultrastructural observation showed a number of TAG-rich plastoglobuli formed in plastids. Conclusion The results suggested that AGPAT1 overexpression could elevate TAG biosynthesis and, moreover, revealed the occurrence of plastidial TAG synthesis in the diatom. Overall, our data provide a new insight into microalgal lipid metabolism and candidate target for metabolic engineering.

The contact angle, as a vital measured parameter of wettability of material surface, has long been in dispute whether it is affected by gravity. Herein, we measured the advancing and receding contact angles on extremely low contact angle hysteresis surfaces under different gravities (1-8G) and found that both of them decrease with the increase of the gravity. The underlying mechanism is revealed to be the contact angle hysteresis and the deformation of the liquid-vapor interface away from the solid surface caused by gradient distribution of the hydrostatic pressure. The real contact angle is not affected by gravity and cannot measured by an optical method. The measured apparent contact angles are angles of inclination of the liquid-vapor interface away from the solid surface. Furthermore, a new equation is proposed based on the balance of forces acting on the three-phase contact region, which quantitatively reveals the relation of the apparent contact angle with the interfacial tensions and gravity. This finding can provide new horizons for solving the debate on whether gravity affects the contact angle and may be useful for the accurate measurement of the contact angle and the development of a new contact angle measurement system.

To speed up the fabrication of optical metamaterials by making use of the fast speed advantage of femtosecond laser preparation, a metamaterial appropriate for femtosecond laser processing was designed, and the interaction between femtosecond laser and metal-dielectric-metal fishnet stacks was investigated in detail. Two kinds of processing mechanisms, thermal melting and stress break, were revealed during the fabrication. The thermal melting process, dominated by the interaction of femtosecond laser with metals, makes the upper and lower metal layers adhere to each other, which leads to the magnetic resonance impossible. The stress break process, dominated by the interaction of femtosecond laser with dielectrics, can keep the upper and lower metal coatings isolated. Fishnet optical metamaterial was fabricated by femtosecond laser-induced stress break technique, using back side ablation, high numerical aperture and super-Gaussian beam. The resolution and speed can reach 500 nm, and 100 units/s, respectively. Spectrophotometer measurement results proved that the magnetic resonances were found in the fishnet nanostructure. The theoretical refractive index of the metamaterial on a glass substrate reached −0.12 at the wavelength of 3225 nm. It proved that femtosecond laser-induced stress break was a good and fast tool during the fabrication of optical metamaterials.

Poor soil structure and nutrients, excessive exchangeable Na+, high pH as well as low enzyme activities are common in the solonetz, and significantly restrict corn (Zea mays L.) production. Cattle manure application combined with deep tillage is an important management practice that can affect soil physico-chemical properties and enzyme activities as well as corn yield in the solonetz. Field experiments were carried out in a randomized complete block design comprising four treatments: Corn with conventional tillage was used as a control, and corn with manure application combined with deep tillage as well as film mulching and aluminium sulfate were used as the experimental treatments, respectively. The relationship between corn yield and measured soil properties was determined using stepwise regression analysis. Manure application combined with deep tillage management was more effective than conventional tillage for increasing corn yield and for improving soil properties in the solonetz. The highest corn yield was obtained in the treatments with manure application+deep tillage+plastic film mulching (11 472 and 12 228 kg ha−1), and increased by 38 and 43% comparing with the control treatment (8 343 and 8 552 kg ha−1) both in the 2013 and 2014 experiments, respectively. Using factor analysis, three factors were obtained, which represented soil fertility status, soil saline-alkaline properties and soil structural properties both in the 2013 and 2014 experiments, respectively. Manure and deep tillage management resulted in two distinct groups of soil properties: (1) soils with manure application combined with deep tillage and (2) soils with conventional tillage. Stepwise regression analysis showed that corn yield was significantly and positively correlated to urease and available P, as well as negatively correlated to pH, electrical conductivity (EC), exchange sodium percentage (ESP), and bulk density (ρb). We concluded that ρb was dominant factor for corn yield on the basis of discriminant coefficient. Manure application combined with deep tillage management resulted in an increase in corn yield mainly owing to improved soil structural properties, followed by decreased soil saline-alkaline obstacle as well as increased urease activity and available P. This result is likely that the improvement in soil organic matter (SOM) from manure application greatly and positively contributed to better soil physico-chemical properties and enzyme activities, especially decrease in ρb. Suggestion for corn production should be improvement in soil structural properties firstly. This could cause decrease in ρb that key factor which limited the corn production in the solonetz.

Several studies have attempted to reduce diagnostic delay and identify biomarkers for drug development in amyotrophic lateral sclerosis (ALS). In this study, we aimed to evaluate the diagnostic accuracy for ALS of cerebrospinal fluid (CSF) neurofilament (Nf), Tau protein, and inflammatory factors such as interleukin (IL)-2, IL-6, IL-10, IL-15, and granulocyte-macrophage colony-stimulating factor (GMCSF) in Chinese patients. Our prospective study measured the concentration of phosphorylated Nf heavy chain (pNfH), Nf light chain (NfL), Tau, pTau, and inflammatory factors in the CSF of 85 patients. Detailed clinical data and laboratory, neuroimaging, and neurophysiological findings were recorded. The concentrations of pNfH and NfL were higher in the ALS group than in the control group. At the 1104 pg/mL pNfH cutoff, the specificity was 68.8%, the sensitivity 100%, and the area under the curve (AUC) 0.907. At the 1,139 pg/mL NfL cutoff, the specificity was 56.3%, the sensitivity 96.2%, and the AUC 0.775. There was no significant difference in the concentrations of Tau, pTau, IL-2, IL-6, IL-10, IL-15, and GMCSF between the ALS and control groups (p > 0.05). In the ALS group, the concentration of pNfH in the CSF was correlated with disease duration (r = −0.475, p < 0.001). This is the first prospective study to confirm the diagnostic value of Nf for ALS in Chinese patients.

ORF3a, a newly identified non-AUG-initiated ORF encoded by members of genera Polerovirus and Luteovirus, is required for long-distance movement in plants. However, the mechanism of action of P3a in viral systemic movement is still not clear. In this study, sequencing of a brassica yellows virus (BrYV) mutant defective in systemic infection revealed two-nucleotide variation at positions 3406 and 3467 in the genome. Subsequent nucleotide substitution analysis proved that only the non-synonymous substitution (C→U) at position 3406, resulting in P3aP18L, abolished the systemic infection of BrYV. Preliminary investigation showed that wild type BrYV was able to load into the petiole of the agroinfiltrated Nicotiana benthamiana leaves, whereas the mutant displayed very low efficiency. Further experiments revealed that the P3a and its mutant P3aP18L localized to the Golgi apparatus and near plasmodesmata, as well as the endoplasmic reticulum. Both P3a and P3aP18L were able to self-interact in vivo, however, the mutant P3aP18L seemed to form more stable dimer than wild type. More interestingly, we confirmed firstly that the ectopic expression of P3a of other poleroviruses and luteoviruses, as well as co-infection with Pea enation mosaic virus 2 (PEMV 2), restored the ability of systemic movement of BrYV P3a defective mutant, indicating that the P3a is functionally conserved in poleroviruses and luteoviruses and is redundant when BrYV co-infects with PEMV 2. These observations provide a novel insight into the conserved function of P3a and its underlying mechanism in the systemic infection.

Amyloid beta (Aß)-induced oxidative stress is a major pathologic hallmark of Alzheimer′s disease. Cyanidin, a natural flavonoid compound, is neuroprotective against oxidative damage-mediated degeneration. However, its molecular mechanism remains unclear. Here, we investigated the effects of cyanidin pretreatment against Aß-induced neurotoxicity in PC12 cells, and explored the underlying mechanisms. Cyanidin pretreatment significantly attenuated Aß-induced cell mortality and morphological changes in PC12 cells. Mechanistically, cyanidin effectively blocked apoptosis induced by Aß, by restoring the mitochondrial membrane potential via upregulation of Bcl-2 protein expression. Moreover, cyanidin markedly protected PC12 cells from Aß-induced DNA damage by blocking reactive oxide species and superoxide accumulation. These results provide evidence that cyanidin suppresses Aß-induced cytotoxicity, by preventing oxidative damage mediated by reactive oxide species, which in turn inhibits mitochondrial apoptosis. Our study demonstrates the therapeutic potential of cyanidin in the prevention of oxidative stress-mediated Aß neurotoxicity.

Polerovirus P0 suppressors of host gene silencing contain a consensus F-box-like motif with Leu/Pro (L/P) requirements for suppressor activity. The Inner Mongolian Potato leafroll virus (PLRV) P0 protein (P0PL-IM) has an unusual F-box-like motif that contains a Trp/Gly (W/G) sequence and an additional GW/WG-like motif (G139/W140/G141) that is lacking in other P0 proteins. We used Agrobacterium infiltration-mediated RNA silencing assays to establish that P0PL-IM has a strong suppressor activity. Mutagenesis experiments demonstrated that the P0PL-IM F-box-like motif encompasses amino acids 76-LPRHLHYECLEWGLLCG THP-95, and that the suppressor activity is abolished by L76A, W87A, or G88A substitution. The suppressor activity is also weakened substantially by mutations within the G139/W140/G141 region and is eliminated by a mutation (F220R) in a C-terminal conserved sequence of P0PL-IM. As has been observed with other P0 proteins, P0PL-IM suppression is correlated with reduced accumulation of the host AGO1-silencing complex protein. However, P0PL-IM fails to bind SKP1, which functions in a proteasome pathway that may be involved in AGO1 degradation. These results suggest that P0PL-IM may suppress RNA silencing by using an alternative pathway to target AGO1 for degradation. Our results help improve our understanding of the molecular mechanisms involved in PLRV infection.

Higher plants exploit posttranscriptional gene silencing as a defense mechanism against virus infection by the RNA degradation system. Plant RNA viruses suppress posttranscriptional gene silencing using their encoded proteins. Three important motifs (F-box-like motif, G139/W140/G141-like motif, and C-terminal conserved region) in P0 of Potato leafroll virus (PLRV) were reported to be essential for suppression of RNA silencing activity. In this study, Agrobacterium-mediated transient experiments were carried out to screen the available amino acid substitutions in the F-box-like motif and G139/W140/G141-like motif that abolished the RNA silencing suppression activity of P0, without disturbing the P1 amino acid sequence. Subsequently, four P0 defective mutants derived from a full-length cDNA clone of PLRV (L76F and W87R substitutions in the F-box-like motif, G139RRR substitution in the G139/W140/G141-like motif, and F220R substitution in the C-terminal conserved region) were successfully generated by reverse PCR and used to investigate the impact of these substitutions on PLRV infectivity. The RT-PCR and western blot analysis revealed that these defective mutants affected virus accumulation in inoculated leaves and systemic movement in Nicotiana benthamiana as well as in its natural hosts, potato and black nightshade. These results further demonstrate that the RNA silencing suppressor of PLRV is required for PLRV accumulation and systemic infection.

Microalgae have been emerging as an important source for the production of bioactive compounds. Marine diatoms can store high amounts of lipid and grow quite quickly. However, the genetic and biochemical characteristics of fatty acid biosynthesis in diatoms remain unclear. Glycerophospholipids are integral as structural and functional components of cellular membranes, as well as precursors of various lipid mediators. In addition, diacylglycerol acyltransferase (DGAT) is a key enzyme that catalyzes the last step of triacylglyceride (TAG) biosynthesis. However, a comprehensive sequence-structure and functional analysis of DGAT in diatoms is lacking. In this study, an isoform of diacylglycerol acyltransferase type 2 of the marine diatom Phaeodactylum tricornutum was characterized. Surprisingly, DGAT2 overexpression in P. tricornutum stimulated more oil bodies, and the neutral lipid content increased by 35%. The fatty acid composition showed a significant increase in the proportion of polyunsaturated fatty acids; in particular, EPA was increased by 76.2%. Moreover, the growth rate of transgenic microalgae remained similar, thereby maintaining a high biomass. Our results suggest that increased DGAT2 expression could alter fatty acid profile in the diatom, and the results thus represent a valuable strategy for polyunsaturated fatty acid production by genetic manipulation.