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Anisodus tanguticus (Maxim.) Pascher is an important Tibetan folk medicine and the source of tropane alkaloids (TAs) grown in Qinghai-Tibet Plateau. There are marked differences in quality of A. tanguticus from geographic areas. The aim of present research was to establish a method for the quantitative analysis of TAs coupled with chemometrics analysis to trace geographical origins. Qualitative analysis of TAs in A. tanguticus was carried out using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and quantitative analysis of TAs in different plant organs from different geographical origin was achieved. Contents of TAs were subjected to the principal component analysis, and orthogonal partial least-squares discriminant analysis. The contents of the three marker compounds (anisodamine, anisodine and atropine) in the roots and acrial parts of A. tanguticus were positive correlated and varied significantly from different geographical origins. Principal component analysis, and orthogonal partial least-squares discriminant analysis results showed excellent discrimination between different geographical origin of A. tanguticus. This study could provide comprehensive evaluation and further utilization of A. tanguticus resources.

ibrio parahaemolyticus is the main cause of foodborne gastroenteritis, which is widely distributed in shellfish and other seafood. Most V. parahaemolyticus are nonpathogenic, and only a few types, such as serotype O3:K6, are pathogenic, which is also the most prevalent strain in Asia. However, the relationship between this serotype and pathogenicity has yet to be established. The waaf gene is located in the O antigen synthesis gene cluster. Thus, we constructed a waaf gene deletion mutant (i.e., △waaf) of wild-type (WT) which isolated from shellfish serotype O3:K6 via chitin-mediated transformation technology. We then constructed the △waaf complementary strain (i.e., C-Δwaaf) via the Escherichia coli S17 λpir strain by conjugation. The basic physiological characteristics, adhesion to Caco2 cells, and pathogenicity of the WT, Δwaaf, and C-Δwaaf strains were compared. Growth curves showed no remarkable differences between the WT and Δwaaf strains. However, the Δwaaf strain non-reactive to O3 antisera and other 12 O-group antisera of V. parahaemolyticus. Moreover, the number of flagella and extracellular polysaccharides decreased, the adhesion decreased, and the pathogenicity weakened. These characteristics of the C-Δwaaf strain were similar to those of the WT strain. These results indicated that the waaf gene is vital to the serotype in V. parahaemolyticus, and changes in O antigen could affect the pathogenicity of this bacterium. This study will be helpful to understand the pathogenic mechanism of V. parahaemolyticus.

Salmonella , a major foodborne pathogen, causes severe gastrointestinal disease in people and animals worldwide. Plasmid-borne mcr-1 , which confers colistin resistance in Salmonella, has significant epidemiological interest for public health safety. Here, we report the first evidence of mcr-1 -mediated colistin resistance in one multidrug-resistant strain，namely 16062 in this study, from 355 Salmonella isolates collected for Jiaxing foodborne pathogen monitoring in Zhejiang Province in 2015-2019. In addition to colistin, 16062 displayed multidrug resistance to various antimicrobials (β-lactams, quinolone, sulfonamide, florfenicol, ampicillin, streptomycin, nalidixic acid, aminoglycoside, and trimethoprim-sulfamethox). The mcr-1 -carrying IncX4 plasmid (p16062-MCR) in this study shares a conserved structure with other mcr -IncX4 plasmids. We found that other antimicrobial-resistance genes ( aac(6')-Ib-cr , aadA1 , aadA2 , aph(3')-Ia , oqxA , oqxB , sul1 , and cmlA1 ) are located on p16062-cmlA, an atypical IncHI2 plasmid, in isolate 16062. This is the first identification of transferable colistin resistance in foodborne Salmonella isolate collected in Jiaxing city, the 5-year monitoring of which revealed limited dissemination. By determining the genetic features of the plasmid vehicle, the characteristics of transferable mcr genes circulating in isolates from Jiaxing are now clearer.

Implant surface topography plays a crucial role in achieving successful implantation. Simple and controllable surface topographical modifications are considered a promising method to accelerate bone osseointegration for biomedical application. Moreover, comprehension of the mechanism between surface topography and cell osteogenic differentiation is vital for manipulation of these processes to promote bone tissue regeneration. In this study, we investigated the effects of implant surfaces with various sized hierarchical microgroove/nanopore topographies on cell adhesion, osteogenesis, and its underlying mechanism both in vitro and in vivo. Our findings reveals that titanium surface with proper sized microgroove/nanopore topography (SLM-1MAH) exhibits the more satisfying adhesive and osteogenic efficiency than the clinically used sand-blasted, large-grit, and acid etching (SLA) surface. The underlying molecular mechanism lies in the activation of the integrin α2-PI3K-Akt signal pathway, where SLM-1MAH surface increased the protein expression of integrin α2 (Itga2), phosphatidylinositol 3-kinase (PI3K), and phosphorylated serine/threonine kinase Akt (p-Akt) to enhance osteogenesis and osseointegration. Furthermore, the SLM-1MAH surface also displays better osseointegration efficiency with stronger bonding strength than that on SLA surface. This work provides a novel strategy for implant surface topography design to improve bone-implant osseointegration.

Versatile nano-theranostic agents are highly desirable for the precision nanomedicine of tumors in recent years. In this work, a multifunctional diagnostic and therapeutic agent Bi2S3-MnO2@BSA-Ce6 (BMBC) was successfully prepared by a combination strategy of facile chemical coupling and biomineralization under mild conditions. These BMBC nanoparticles with an average diameter of 17.6 ± 4.6 nm exhibited favorable physiological stability and negligible toxicity. This agent expressed the outstanding performance of photothermal conversion (η = 45.8%) under 808 nm laser radiation and generating 1O2 under 650 nm laser irradiation. Due to the strong X-ray attenuation performance of element Bi and the highly effective magnetic moment of Mn2+, this agent exhibited favorable performance of X-ray computed tomography (CT) imaging (35.60 HU•mL•mg−1) and magnetic resonance (MR) imaging (r1 = 5.22 mM−1•s−1) in vitro. Furthermore, most of the HeLa cells treated with BMBC NPs were dead in the MTT and Calcein-AM/PI staining experiments, demonstrating the excellent effect of intracellular synergistic therapy. Overall, this theranostic agent could serve as a promising nano-platform for photothermal/photodynamic synergistic therapy of tumors under the guidance of infrared thermal (IRT)/MR/CT imaging.

This study was designed to evaluate the gastroprotective activity of cirsilineol in hydrochloric acid (HCl)/ethanol-induced gastric ulcer model. Cirsilineol was administered at the doses of 20 and 40 mg/kg in HCl/ethanol-induced rats. The gastroprotective ability was verified by determining the ulcer score, total acidity, hemoglobin, inflammatory cytokines, lipid peroxides, and enzymatic antioxidants superoxide dismutase (SOD) and catalase (CAT) in gastric tissue and serum biochemical analysis. The results showed a favorable increase in the hemoglobin level, antioxidant enzymes (SOD and CAT), restored electrochemical balance (carbon dioxide & anion gap) while a noticeable decrease in ulcer index, total acidity, lipid peroxides, inflammatory cytokines (interleukin-1 beta [IL-1β], IL-6, and tumor necrosis factor alpha) in rats treated with the cirsilineol. The serum biochemical analysis on liver markers (alkaline phosphatases, alanine aminotransferase, and aspartate aminotransferase), kidney markers (urea, creatinine, albumin, globulin, total protein), and lipid profile (triglyceride, high-density lipoprotein, total cholesterol) were attenuated by cirsilineol treatment in rats. Histopathology showed enhanced gastric protection and preserved the integrity of gastric mucosa upon cirsilineol administration. These results ultimately suggest that cirsilineol has gastroprotective effects that prevent the development of gastric ulcer.

Drought is one of the most critical abiotic stresses influencing maize yield. Improving maize cultivars with drought tolerance using marker-assisted selection requires a better understanding of its genetic basis. In this study, a doubled haploid (DH) population consisting of 217 lines was created by crossing the inbred lines Han 21 (drought-tolerant) and Ye 478 (drought-sensitive). The population was genotyped with a 6 K SNP assay and 756 SNP (single nucleotide polymorphism) markers were used to construct a linkage map with a length of 1344 cM. Grain yield (GY), ear setting percentage (ESP), and anthesis–silking interval (ASI) were recorded in seven environments under well-watered (WW) and water-stressed (WS) regimes. High phenotypic variation was observed for all traits under both water regimes. Using the LSMEAN (least-squares mean) values from all environments for each trait, 18 QTL were detected, with 9 associated with the WW and 9 with the WS regime. Four chromosome regions, Chr. 3: 219.8–223.7 Mb, Chr. 5: 191.5–194.7 Mb, Chr. 7: 132.2–135.6 Mb, and Chr. 10: 88.2–89.4 Mb, harbored at least 2 QTL in each region, and QTL co-located in a region inherited favorable alleles from the same parent. A set of 64 drought-tolerant BC3F6 lines showed preferential accumulation of the favorable alleles in these four regions, supporting an association between the four regions and maize drought tolerance. QTL-by-environment interaction analysis revealed 28 edQTL (environment-dependent QTL) associated with the WS regime and 22 associated with the WW regime for GY, ESP, and ASI. All WS QTL and 55.6% of WW QTL were located in the edQTL regions. The hotspot genomic regions identified in this work will support further fine mapping and marker-assisted breeding of drought-tolerant maize.

To clarify the histopathological characteristics of Camellia oleifera that were infected by Colletotrichum fructicola, this study utilized frozen tissue sections, whole-stain clearing and transmission electron microscopy to observe the time of transformation from biotrophic to necrotrophic when Co. fructicola infects leaves, as well as the colonization and expansion of the pathogenic fungus at different stages on the leaves. The Co. fructicola conidia produced germ tubes 2 h post-inoculation, and appressoria began to form 4 h after inoculation. There were two ways that pathogens can use to invade the leaves of Ca. oleifera. One was to directly penetrate the host cuticle, while the other involved invasion from the stomata of the host using germ tubes or hyphae. A few infection vesicles could then be observed in the leaf tissue, and the infection vesicles produce thicker septal primary hyphae after 24 h of infection. When the infection reached the 40 h point, the fungus started to produce secondary hyphae with smaller diameter than that of the primary hyphae and gradually expanded into the adjacent cells. Transmission electron microscopy revealed that when the secondary hyphae entered the mesophyll cells, the contents in the cells were digested and a cavity gradually formed, indicating that the cells had been killed. Thus, the appearance of the secondary hyphae marked the point at which Co. fructicola entered into a destructive necrotrophic stage.

Intron splicing is an essential event in post-transcriptional RNA processing in plant mitochondria, which requires the participation of diverse nuclear-encoded splicing factors. However, it is presently unclear how these proteins cooperatively take part in the splicing of specific introns. In this study, we characterized a nuclear-encoded mitochondrial P-type pentatricopeptide repeat (PPR) protein named EMP603. This protein is essential for splicing of intron 2 in the Nad1 gene and interacts with the mitochondria-localized DEAD-box RNA helicase PMH2-5140, the RAD52-like proteins ODB1-0814 and ODB1-5061, and the CRM domain-containing protein Zm-mCSF1. Further study revealed that the N-terminal region of EMP603 interacts with the DEAD-box of PMH2-5140, the CRM domain of Zm-mCSF1, and OBD1-5061, but not with OBD1-0814, whereas the PPR domain of EMP603 can interact with ODB1-0814, ODB1-5061, and PMH2-5140, but not with Zm-mCSF1. Defects in EMP603 severely disrupt the assembly and activity of mitochondrial complex I, leading to impaired mitochondrial function, and delayed seed development. The interactions revealed between EMP603 and PMH2-5140, ODB1-0814, ODB1-5061, and Zm-mCSF1 indicate a possible involvement of a dynamic ‘spliceosome-like’ complex in intron splicing, and may accelerate the elucidation of the intron splicing mechanism in plant mitochondria.

Breast cancer is one of the most common and heterogeneous malignancies. Although the prognosis of breast cancer has improved with the development of early screening, the mechanisms underlying tumorigenesis and progression remain incompletely understood. DNA methylation has been implicated in tumorigenesis and tumor development and, in the present study. we screened methylation‐driven genes and explored their prognostic values in breast cancer. RNA‐sequencing (RNA‐Seq) transcriptome data and DNA methylation data of the TCGA‐BRCA dataset were obtained from The Cancer Genome Atlas. Differentially expressed genes and differentially methylated genes were identified separately. The intersected 783 samples with both RNA‐Seq data and DNA methylation data were selected for further analysis. Fifty‐six methylation‐driven genes were identified using the MethylMix r package and 10 prognosis methylation‐driven genes (CDO1, CELF2, ITPAIPL1, KCNH8, PTK6, RAB25, RIC3, USP44, ZSCAN1 and ZSCAN23) were further screened by combined methylation and gene expression analysis. Based on the methylation data of the screened 10 methylation‐driven genes, six subgroups were identified with the ConsensusClusterPlus r package. The protein levels of the 10 prognostic methylation‐driven genes were detected by immunohistochemical experiments. Moreover, based on the RNA‐Seq data, a signature calculating the risk score of each patient was developed with stepwise regression. The risk score and other clinical features (age and stage) were confirmed to be independent prognostic factors by univariate and multivariate Cox regression analyses. Finally, a prognostic nomogram incorporating all the significant factors was integrated to predict the 3‐, 5‐ and 7‐year overall survival. Taken together, the methylation‐driven genes identified here may be potential biomarkers of breast cancer., DNA methylation has been implicated in tumorigenesis and tumor development. In the present study, we screened methylation‐driven genes and explored their prognostic values in breast cancer by combined methylation and gene expression analysis. The methylation‐driven genes identified may be potential biomarkers of breast cancer.

Background We aimed to assess the efficacy of different initial intravenous immunoglobulin (IVIG) regimens in Kawasaki disease (KD) patients to find more cost‐effective therapy options. Methods A multicenter, open‐label, blind‐endpoint randomized controlled trial was conducted from January 2014 to December 2015. Patients with KD, within 10 days of illness, were randomly assigned to receive different IVIG regimens (Group A, 2 g/kg once; Group B, 1 g/kg for 2 consecutive days; Group C, 1 g/kg once) and aspirin 30mg/kg/d. Primary outcomes included hours to defervescence and development of coronary artery lesions during the study period. Major secondary outcomes included total fever days, total dose of IVIG, changes of laboratory data, length of stay, and hospitalization expenses. (ClinicalTrials.gov: NCT02439996). Results A total of 404 patients underwent randomization. No difference was found in the outcomes of defervescence among three groups at 6, 12, 24, and 36 hours after completion of initial IVIG infusion. There were no differences in the incidence of coronary artery lesions during the study period (at week 2, month 1, month 3, and month 6 of illness), changes of laboratory data, total fever days, and length of stay. Group C patients had the lowest total dose of IVIG (mean: 1.2 vs 2.2 vs 2.1 g/kg; P < 0.001) and hospitalization expenses (mean: 8443.8 vs 10798.4 vs 11011.4 Chinese Yuan; P < 0.001) than other two groups. Conclusions A single dose of 1g/kg IVIG is a low‐cost treatment with the same efficacy as 2 g/kg IVIG and can be an option for the initial therapy of KD patients.

Endothelial cell dysfunction plays a fundamental role in the pathogenesis of atherosclerosis (AS), and endothelial autophagy has protective effects on the development of AS. Our previous study had shown that oxidized low-density lipoprotein/β2-glycoprotein I/anti-β2-glycoprotein I antibody (oxLDL/β2GPI/anti-β2GPI) complex could promote the expressions of inflammatory cytokines and enhance the adhesion of leukocytes to endothelial cells. In the present study, we aimed to assess the effects of oxLDL/β2GPI/anti-β2GPI complex on endothelial autophagy and explore the associated potential mechanisms. Human umbilical vein endothelial cells (HUVECs) and mouse brain endothelial cell line (bEnd.3) were used as models of the vascular endothelial cells. Autophagy was evaluated by examining the expressions of autophagic proteins using western blotting analysis, autophagosome accumulation using transmission electron microscopy, and RFP-GFP-LC3 adenoviral transfection and autophagic flux using lysosome inhibitor chloroquine. The expressions of phospho-PI3K, phospho-AKT, phospho-mTOR, and phospho-eNOS were determined by western blotting analysis. 3-Methyladenine (3-MA) and rapamycin were used to determine the role of autophagy in oxLDL/β2GPI/anti-β2GPI complex-induced endothelial cell dysfunction. We showed that oxLDL/β2GPI/anti-β2GPI complex suppressed the autophagy, evidenced by an increase in p62 protein, a decrease in LC3-II and Beclin1, and a reduction of autophagosome generation in endothelial cells. Moreover, inhibition of autophagy was associated with PI3K/AKT/mTOR and eNOS signaling pathways. Rapamycin attenuated oxLDL/β2GPI/anti-β2GPI complex-induced endothelial inflammation, oxidative stress, and apoptosis, whereas 3-MA alone induced the endothelial injury. Our results suggested that oxLDL/β2GPI/anti-β2GPI complex inhibited endothelial autophagy via PI3K/AKT/mTOR and eNOS signaling pathways and further contributed to endothelial cell dysfunction. Collectively, our findings provided a novel mechanism for vascular endothelial injury in AS patients with an antiphospholipid syndrome (APS) background.

Summary Our study reports a novel dominant COL1A1 mutation in OI. Using a zebrafish model, we confirmed that the glycine to serine substitution at position 608 of the COL1A1 protein has deleterious effects on bone development. Purpose Osteogenesis imperfecta (OI), also known as brittle bone disease, is a group of genetic disorders. Mutations in two genes, collagen type I alpha 1 chain (COL1A1) and collagen type I alpha 1 chain (COL1A2), which encode the pro-a1 (I) and pro-a2 (I) chains of type I collagen, respectively, the most abundant form of collagen in the human body, cause most cases of OI. Methods In this study, we used panel-based next-generation sequencing for prenatal diagnosis of a fetus whose ultrasound images suggested OI. A de novo mutation in COL1A1 gene was suspected to cause the phenotype. To validate the effect of this mutation in a zebrafish model, we constructed a plasmid containing the corresponding mutation in the zebrafish gene col1a1a (c.1744 G > A), and overexpressed the mutant protein in zebrafish larvae. Results We identified a novel COL1A1 mutation (c.1822 G > A; p.Gly608Ser) in the fetus but not in her parents by an skeletal dysplasias panel. Bioinformatic analysis showed that the affected residue (p.Gly608) is highly conserved from zebrafish to humans. In contrast to larvae expressing wild-type (WT) col1a1a and enhanced green fluorescent protein (EGFP), col1a1a mutation-expressing larvae showed significant spinal curvature and embryonic lethality, mimicking the phenotype of human OI. Conclusions Our study revealed the pathogenicity of a de novo mutation, c.1822 G > A, in human COL1A1, which expands the mutation spectrum of OI.

Carbon monoxide (CO) is an important gaseous signaling molecule. The use of CO-releasing molecules such as metal carbonyls enables the elucidation of the pleiotropic functions of CO. Although metal carbonyls show a broad-spectrum antimicrobial activity, it remains unclear whether the bactericidal property originates from the transition metals or the released CO. Here, we develop nonmetallic CO-releasing micelles via a photooxygenation mechanism of 3-hydroxyflavone derivatives, enabling CO release under red light irradiation (e.g., 650 nm). Unlike metal carbonyls that non-specifically internalize into both Gram-positive and Gram-negative bacteria, the nonmetallic micelles are selectively taken up by S. aureus instead of E. coli cells, exerting a selective bactericidal effect. Further, we demonstrate that the CO-releasing micelles can cure methicillin-resistant S. aureus (MRSA)-infected wounds, simultaneously eradicating MRSA pathogens and accelerating wound healing.

RUBylation plays essential roles in plant growth and development through regulating Cullin-RING ubiquitin E3 ligase (CRL) activities and the CRL-mediated protein degradations. However, the function of RUBylation in regulating kernel development remains unclear. Through genetic and molecular analyses of a small kernel 501 (smk501) mutant in maize (Zea mays L.), we cloned the smk501 gene, revealed its molecular function, and defined its roles in RUBylation pathway and seed development. Smk501 encodes a RUBylation activating enzyme E1 subunit ZmECR1 (E1 C-TERMINAL RELATED 1) protein. Destruction in RUBylation by smk501 mutation resulted in less embryo and endosperm cell number and smaller kernel size. The transcriptome and proteome profiling, hormone evaluation and cell proliferation observation revealed that disturbing ZmECR1 expression mainly affects pathways on hormone signal transduction, cell cycle progression and starch accumulation during kernel development. In addition, mutant in zmaxr1 (Auxin resistant 1), another RUB E1 subunit, also showed similar defects in kernel development. Double mutation of zmecr1 and zmaxr1 lead to empty pericarp kernel phenotype. RUBylation is a novel regulatory pathway affecting maize kernel development, majorly through its functions in modifying multiple cellular progresses.

Prioritizing genes for translation to therapeutics for common diseases has been challenging. Here, we propose an approach to identify drug targets with high probability of success by focusing on genes with both gain of function (GoF) and loss of function (LoF) mutations associated with opposing effects on phenotype (Bidirectional Effect Selected Targets, BEST). We find 98 BEST genes for a variety of indications. Drugs targeting those genes are 3.8-fold more likely to be approved than non-BEST genes. We focus on five genes (IGF1R, NPPC, NPR2, FGFR3, and SHOX) with evidence for bidirectional effects on stature. Rare protein-altering variants in those genes result in significantly increased risk for idiopathic short stature (ISS) (OR = 2.75, p = 3.99 × 10−8). Finally, using functional experiments, we demonstrate that adding an exogenous CNP analog (encoded by NPPC) rescues the phenotype, thus validating its potential as a therapeutic treatment for ISS. Our results show the value of looking for bidirectional effects to identify and validate drug targets., Prioritising genes as potential drug targets is challenging and often unsuccessful once testing efficacy in humans. Here, the authors propose an approach to identifying drug targets that uses evidence from gain- or loss-of-function mutations associated with bidirectional effects on phenotypes.

The insulin signalling pathway is one of the most studied pathways, including ovary maturation and female reproduction. However, downstream elements involved in this cascade remain unknown. Here, we identified and characterized an insulin-like receptor (IR) in female Macrobrachium rosenbergii, and named as Mro-IR. The deduced Mro-IR contained conserved domains of IR proteins, including two ligand-binding domains, a furin-like domain, two fibronectin-3 domains, a transmembrane domain and an intracellular tyrosine kinase domain in order. Multiple sequence alignment of the first ligand-binding domain and the tyrosine kinase domain in Mro-IR revealed a high degree of similarity to other representative IRs. Mro-IR was clustered into a separate branch with IRs from decapods by phylogenetic analysis. Additionally, tissue distribution analysis showed that Mro-IR was uniquely expressed in the ovary of M. rosenbergii. This study would possibly provide a new highlight to the molecular mechanism of insulin-like signalling cascade, regulating female sexual differentiation in crustaceans.

Rice accumulates higher levels of cadmium (Cd) in grain than other cereals, and hybrid rice varieties cover around half of the total rice area in China. Therefore, understanding the phenotypic and genetic characteristics of Cd accumulation in rice hybrids is vital for food safety. An incomplete diallel cross design was used to analyse the heritability and combining ability of Cd accumulation in rice hybrids growing in soil contaminated with Cd. The quantitative real-time PCR was used to detect the gene expressions that related to Cd uptake and transport. The Cd contents in brown rice, nodes and roots were detected by ICPOES, and the Cd contents in xylem sap were detected by ICPMS. Additive effects dominated the Cd accumulation in brown rice, upper nodes, and the Cd translocation efficiency from roots to nodes of hybrids. The Cd accumulation in brown rice was positively correlated with Cd accumulation in upper nodes and translocation efficiency from roots to nodes. Regarding the general combining ability, the restorer lines R2292 and R2265, and the male sterile lines KeBX58S, Ke20A, Ke76-1BS and Ke244-1BS were found to be potential elite parental lines for developing hybrid rice with low Cd accumulation. Hybrid rice with low Cd accumulation in brown rice can be derived from parental lines with low general combining ability for Cd content in brown rice.

TianWei Guo,1,* Wei Wang,2,* YueXia Ji,1 Min Zhang,3 GuoYing Xu,4 Sen Lin5 1Department of Pathology, Changshu Hospital Affiliated to Nanjing University of Chinese Medicine, Changshu, Jiangsu, People’s Republic of China; 2Department of Pathology, The Affiliated Obstetrics and Gynecology Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing, Jiangsu, People’s Republic of China; 3Department of Pathology, Children’s Hospital Affiliated to Soochow University, Suzhou, Jiangsu, People’s Republic of China; 4School of Medical Technology, Jiangsu College of Nursing, Huai’an, Jiangsu, People’s Republic of China; 5The Affiliated Huai’an Hospital of Xuzhou Medical University and the Second People’s Hospital of Huai’an, Huai’an, Jiangsu, People’s Republic of China*These authors contributed equally to this workCorrespondence: Sen LinThe Affiliated Huai’an Hospital of Xuzhou Medical University and the Second People’s Hospital of Huai’an, Huai’an, Jiangsu, People’s Republic of ChinaEmail linsen378@163.comGuoYing XuSchool of Medical Technology, Jiangsu College of Nursing, Huai’an, Jiangsu, People’s Republic of ChinaEmail xuguoying3@163.comIntroduction: Growing evidences imply that multiple long non-coding RNAs (lncRNAs) play a significant role in the treatment of cancer. Therefore, it is of great significance to discover new biomarkers or therapeutic targets of gastric cancer (GC). However, the potential molecular mechanism of lncPROX1-AS1 in GC remains unknown. The objective of current study is to investigate the effect of PROX1-AS1 in GC.Methods: Thus, we detect that PROX1-AS1 is over-expressed in tissues and cell lines of GC using qRT-PCR analysis. CCK-8, colony formation, flow cytometry, wounding healing and transwell analyses were performed to explore the effect of PROX1-AS1 on GC malignant behaviors.Results: It is further disclosed that silencing of PROX1-AS1 represses cell proliferation, migration, and invasion, whereas promotes cell apoptosis in GC. Bioinformatics analysis suggests that miR-877-5p is negatively regulated by PROX1-AS1 and ectopic of miR-877-5p alleviates the malignant behaviors of GC. Subsequently, miR-877-5p suppresses the activity of PD-L1-3ʹ UTR. At last, rescue assays demonstrated that the GC progression is suppressed by sh-PROX1-AS1 and facilitated on account of miR-877-5p inhibitors and then is retrieved by sh-PD-L1.Discussion: Our findings reveal that PROX1-AS1 exerts its role via miR-877-5p/PD-L1 axis in the GC progression, suggesting that PROX1-AS1 may represent a new therapeutic target for the diagnosis and treatment of GC patients.Keywords: PROX1-AS1, miR-877-5p, PD-L1, proliferation, apoptosis, gastric cancer

Background and Aims Knowledge of plant resource acquisition strategies is crucial for understanding the mechanisms mediating the responses of ecosystems to external nitrogen (N) input. However, few studies have considered the joint effects of above-ground (light) and below-ground (nutrient) resource acquisition strategies in regulating plant species responses to N enrichment. Here, we quantified the effects of light and non-N nutrient acquisition capacities on species relative abundance in the case of extra N input. Methods Based on an N-manipulation experiment in a Tibetan alpine steppe, we determined the responses of species relative abundances and light and nutrient acquisition capacities to N enrichment for two species with different resource acquisition strategies (the taller Stipa purpurea, which is colonized by arbuscular mycorrhizal fungi, and the shorter Carex stenophylloides, which has cluster roots). Structural equation models were developed to explore the relative effects of light and nutrient acquisition on species relative abundance along the N addition gradient. Key Results We found that the relative abundance of taller S. purpurea increased with the improved light acquisition along the N addition gradient. In contrast, the shorter C. stenophylloides, with cluster roots, excelled in acquiring phosphorus (P) so as to elevate its leaf P concentration under N enrichment by producing large amounts of carboxylate exudates that mobilized moderately labile and recalcitrant soil P forms. The increased leaf P concentration of C. stenophylloides enhanced its light use efficiency and promoted its relative abundance even in the shade of taller competitors. Conclusions Our findings highlight that the combined effects of above-ground (light) and below-ground (nutrient) resources rather than light alone (the prevailing perspective) determine the responses of grassland community structure to N enrichment.

A new two-step enzymatic conversion process for the production of a novel mono-α-1,4-glucosylated rebaudioside A derivative (RA1G) was established via transglycosylation followed by hydrolyzation. In the transglycosylation reaction catalyzed by cyclodextrin glycosyltransferase, rebaudioside A (RA) was converted into glucosylated RA derivatives (RAGs), and the maximum conversion of 87.8% was obtained with the optimal conditions of 2 U/mL enzyme, 82.5 mg/mL β-cyclodextrin, and 82.5 mg/mL RA at 60 °C for 5 h. The obtained RAG solution was then directly used in hydrolyzation. Four amylases were screened for shortening the newly synthesized α-glucans of RAGs. A glucoamylase was found to produce RA1G as the single glucosylated product, and the maximum yield of 53.3% was achieved with the optimal conditions of adding 1.5 U/mL glucoamylase into RAG solution at 60 °C for 3 h. RA1G was identified as 13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl) oxy] ent-kaur-16-en-19-oic acid-[(4-O-α-D-glucopyranosyl-β-D-glucopyranosyl) ester] by MS and NMR analysis and showed an improved sensory quality compared to RA.

Coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 has caused a global pandemics. To facilitate the detection of SARS-CoV-2 infection, various RT-LAMP assays using 19 sets of primers had been developed, but never been compared. We performed comparative evaluation of the 19 sets of primers using 4 RNA standards and 29 clinical samples from COVID-19 patients. Six of 15 sets of primers were firstly identified to have faster amplification when tested with four RNA standards, and were further subjected to parallel comparison with the remaining four primer sets using 29 clinical samples. Among these 10 primer sets, Set-4 had the highest positive detection rate of SARS-CoV-2 (82.8%), followed by Set-10, Set-11, and Set-13 and Set-17 (75.9%). Set-14 showed the fastest amplification speed (Tt value Nsp3, S, S, E, N and N gene regions of SARS-CoV-2, respectively, were determined to be better than the other primer sets. Two RT-LAMP assays with the Set-4 primers in combination with any one of four other primer sets (Set-14, Set-10, Set-11, and Set-13) were recommended to be used in the COVID-19 surveillance.

Background Rheum tanguticum (Rh. tanguticum) is an important traditional Chinese medicine plant, “Dahuang”, which contains productive metabolites and occupies wide habitats on the Qinghai-Tibet plateau. Plants occupying wide habitats usually vary in phenotypes such as in morphology and metabolism, thereby developing into different ecotypes. Under canopy and open habitats are a pair of dissimilar habitats which possess Rh. tanguticum plants. However, few studies have focused on the effect of habitats on Rh. tanguticum growth, particularly combining morphological and metabolic changes. This study focused on Rh. tanguticum plants growing in under canopy and open habitats where morphology and metabolism changes were quantified using non-target metabolism methods. Results The obtained results indicated that the two dissimilar habitats led to Rh. tanguticum developing into two distinct ecotypes where the morphology and metabolism were simultaneously changed. Under canopy habitats bred morphologically smaller Rh. tanguticum plants which had a higher level of metabolites (22 out of 31) which included five flavonoids, four isoflavonoids, and three anthracenes. On the other hand, the open habitats produced morphologically larger Rh. tanguticum plants having a higher level of metabolites (9 out of 31) including four flavonoids. 6 of the 31 metabolites were predicted to have effect targets, include 4 represent for under canopy habitats and 2 for open habitats. Totally, 208 targets were connected, among which 42 were communal targets for both under canopy and open habitats represent compounds, and 100 and 66 were unique targets for under canopy superior compounds and open habitats superior compounds, respectively. In addition, aloe-emodin, emodin, chrysophanol, physcion, sennoside A and sennoside B were all more accumulated in under canopy habitats, and among which aloe-emodin, emodin, chrysophanol and physcion were significantly higher in under canopy habitats. Conclusions This study determined that Rh. tanguticum growing in under canopy and in open habitats developed into two distinct ecotypes with morphological and metabolic differences. Results of network pharmacology study has indicated that “Dahuang” coming from different habitats, such as under canopy and open habitats, are different in effect targets and thus may have different medicinal use. According to target metabolomics, under canopy habitats may grow better “Dahuang”.

Splicing of plant organellar group II introns from precursor-RNA transcripts requires the assistance of nuclear-encoded splicing factors. Maturase (nMAT) is one such factor, as its three homologs (nMAT1, 2 and 4) have been identified as being required for the splicing of various mitochondrial introns in Arabidopsis. However, the function of nMAT in maize (Zea mays L.) is unknown. In this study, we identified a seed development mutant, empty pericarp 2441 (emp2441) from maize, which showed severely arrested embryogenesis and endosperm development. Positional cloning and transgenic complementation assays revealed that Emp2441 encodes a maturase-related protein, ZmnMAT3. ZmnMAT3 is highly expressed during seed development and its protein locates to the mitochondria. The loss of function of ZmnMAT3 resulted in the reduced splicing efficiency of various mitochondrial group II introns, particularly of the trans-splicing of nad1 introns 1, 3 and 4, which consequently abolished the transcript of nad1 and severely impaired the assembly and activity of mitochondrial complex I. Moreover, the Zmnmat3 mutant showed defective mitochondrial structure and exhibited expression and activity of alternative oxidases. These results indicate that ZmnMAT3 is essential for mitochondrial complex I assembly during kernel development in maize.

The RNA-binding protein QKI belongs to the hnRNP K-homology domain protein family, a well-known regulator of pre-mRNA alternative splicing and is associated with several neurodevelopmental disorders. Qki is found highly expressed in developing and adult hearts. By employing the human embryonic stem cell (hESC) to cardiomyocyte differentiation system and generating QKI-deficient hESCs (hESCs-QKIdel) using CRISPR/Cas9 gene editing technology, we analyze the physiological role of QKI in cardiomyocyte differentiation, maturation, and contractile function. hESCs-QKIdel largely maintain normal pluripotency and normal differentiation potential for the generation of early cardiogenic progenitors, but they fail to transition into functional cardiomyocytes. In this work, by using a series of transcriptomic, cell and biochemical analyses, and the Qki-deficient mouse model, we demonstrate that QKI is indispensable to cardiac sarcomerogenesis and cardiac function through its regulation of alternative splicing in genes involved in Z-disc formation and contractile physiology, suggesting that QKI is associated with the pathogenesis of certain forms of cardiomyopathies., RNA binding protein Quaking (QKI) is known for its broad function in pre-mRNA splicing and modification and its association with several neurodevelopmental disorders. Here the authors reveal that QKI-mediated regulation of RNA splicing is indispensable to cardiac development and contractile physiology.

Emergence of antibiotic resistance presents a major setback to global health, and shortage of antibiotic pipelines has created an urgent need for development of alternative therapeutic strategies. Bacteriophage (phage) therapy is considered as a potential approach for treatment of the increasing number of antibiotic-resistant pathogens. Phage-antibiotic synergy (PAS) refers to sublethal concentrations of certain antibiotics that enhance release of progeny phages from bacterial cells. A combination of phages and antibiotics is a promising strategy to reduce the dose of antibiotics and the development of antibiotic resistance during treatment. In this review, we highlight the state-of-the-art advancements of PAS studies, including the analysis of bacterial-killing enhancement, bacterial resistance reduction, and anti-biofilm effect, at both in vitro and in vivo levels. A comprehensive review of the genetic and molecular mechanisms of phage antibiotic synergy is provided, and synthetic biology approaches used to engineer phages, and design novel therapies and diagnostic tools are discussed. In addition, the role of engineered phages in reducing pathogenicity of bacteria is explored.

Background & Aims Cancer-associated fibroblasts (CAFs) play a key role in the cancer process, but the research progress is hampered by the paucity of preclinical models that are essential for mechanistic dissection of cancer cell–CAF interactions. Here, we aimed to establish 3-dimensional (3D) organotypic co-cultures of primary liver tumor–derived organoids with CAFs, and to understand their interactions and the response to treatment. Methods Liver tumor organoids and CAFs were cultured from murine and human primary liver tumors. 3D co-culture models of tumor organoids with CAFs and Transwell culture systems were established in vitro. A xenograft model was used to investigate the cell–cell interactions in vivo. Gene expression analysis of CAF markers in our hepatocellular carcinoma cohort and an online liver cancer database indicated the clinical relevance of CAFs. Results To functionally investigate the interactions of liver cancer cells with CAFs, we successfully established murine and human 3D co-culture models of liver tumor organoids with CAFs. CAFs promoted tumor organoid growth in co-culture with direct cell–cell contact and in a Transwell system via paracrine signaling. Vice versa, cancer cells secrete paracrine factors regulating CAF physiology. Co-transplantation of CAFs with liver tumor organoids of mouse or human origin promoted tumor growth in xenograft models. Moreover, tumor organoids conferred resistance to clinically used anticancer drugs including sorafenib, regorafenib, and 5-fluorouracil in the presence of CAFs, or the conditioned medium of CAFs. Conclusions We successfully established murine and human 3D co-culture models and have shown robust effects of CAFs in liver cancer nurturing and treatment resistance., Graphical abstract

Chrysanthemum indici flos (CIF), a traditional functional beverage, has been used in China for thousands of years. However, research on the antioxidant properties and compounds in CIF is insufficient. In the current study, an online gradient extraction (OGE) coupled with a high performance liquid chromatography-ferric reducing antioxidant power (HPLC-FRAP) system for rapid identification of antioxidants in CIF was proposed. The CIF sample (0.3 mg) was extracted online by a gradient mobile phase (acetonitrile and 0.1% formic acid), separated on a Poroshell 120 SB-Aq column, and then detected for the anti-oxidative activity with the online FRAP assay. 36 peaks were found and 28 compounds were identified. Among them, 18 peaks showed antioxidant activity, six of which were detected in CIF for the first time. The total analytical time was less than 45 min, including sample extraction, separation, and antioxidant identification. The results indicate that the OGE-HPLC-FRAP system is a simple, efficient and rapid tool for screening antioxidants in CIF, which provides the scientific data of CIF's antioxidant research and is helpful to improve the quality evaluation of CIF.

Oxidative stress is one of the critical factors that influence rotator cuff healing. Se has been proven to be effective for oxidative stress inhibition and inflammation control in previous studies. This makes it an ideal tool for improving rotator cuff healing. In this work, New Zealand white rabbit rotator cuff healing models were established, and sustained release of Se from porous Se@SiO2 nanocomposites was applied via intraarticular injection. Samples of tendon-bone junctions were removed for histological observation at the 4th and 8th weeks after the operation. The functional proteins collagen II and aggrecan and related inflammatory factors, such as TNF-α, IL-1β and IL-6, were evaluated separately by western blotting (WB) and ELISA. The results indicated that porous Se@SiO2 nanocomposites can significantly reduce scar hyperplasia and fat infiltration, optimize the structural remodelling of collagen fibres and cartilage cells, improve the expression of collagen II and aggrecan and reduce the expression of inflammatory factors. Porous Se@SiO2 nanocomposites showed effective results in promoting rotator cuff healing after surgical repair and had great potential in relevant clinical applications.

In this study, UiO-66-NH2 metal-organic framework (MOF) nanoparticles with peroxidase and oxidase mimetic activities were incorporated into a chitosan (CS) matrix by a simple and environmentally friendly method. The UiO-66-NH2/CS composite membrane possesses the peroxidase mimicking activity in the presence of traces of H2O2, thus resulting in good antibacterial properties. Intriguingly, 30 min of UV pre-irradiation of the UiO-66-NH2/CS composite membrane, in the absence of H2O2, still leads to a good antibacterial activity. This was attributed to the oxidase mimetic activity and the peroxidase mimicking activity of UiO-66-NH2. In such a way, the side effects of direct exposure to UV irradiation and H2O2 can be avoided for wound-healing treatments. The antibacterial mechanism was further proved by antibacterial experiments, TMB·2HCl color development experiments, reactive oxygen species generation tests and electron spin resonance tests. As a potential medical antibacterial dressing, in vitro membranes were also investigated.

Oxidative stress plays a central role in the common pathophysiology of neurodegenerative diseases such as Alzheimer's disease, amyotrophic lateral sclerosis, and Parkinson's disease. Antioxidant therapy has been suggested for the prevention and treatment of neurodegenerative diseases. Compounds derived from natural sources may offer the potential for new treatment options. Semen Celosiae is a traditional Chinese edible herbal medicine with a long history in China and exhibits wide-reaching biological activities such as hepatoprotective, anti-tumor, anti-diarrheal, anti-diabetic, anti-oxidant, etc. In this study, nine saponins and two phenylacetonitrile glycosides were isolated from Semen Celosiae and their structures were identified using ESI-MS and NMR techniques. Among them, compounds 1 and 2 have not been previously reported. The total concentrations of the five triterpenoid saponins and the two phenylacetonitrile glycosides were 3.348 mg g−1 and 0.187 mg g−1, respectively, suggesting that Semen Celosiae is a novel viable source of the two kinds of compounds. These compounds were observed to significantly attenuate t-BHP-induced neuronal damage by effectively enhancing cell viability and decreasing reactive oxygen species generation and cell apoptosis rate in NSC-34 cells. Furthermore, compounds 1 and 7 reduced the ratios of cleaved caspase-3: caspase-3 and cleaved caspase-7: caspase-7 and the level of cytochrome C, while they increased the levels of SOD1 and Beclin 1. These findings suggest that compounds 1–11 are potent inhibitors of neuron injury elicited by t-BHP, possibly via inhibition of oxidative stress and apoptosis, and activation of autophagy; therefore they may be valuable leads for future therapeutic development.

The emergence of multidrug resistance (MDR) in Proteus mirabilis clinical isolates is a growing public health concern and has serious implications for wildlife. What is the role of wildlife has been become one of the hot issues in disseminating antimicrobial resistance (AMR). Here, fifty-four P.mirabilis isolates from 12 different species were identified. Among them, 25 isolates were determined to be MDR by profile of antimicrobial susceptibility, 10 MDR P.mirabilis isolates were subjected to comparative genomic analysis by whole genome sequencing (WGS). Comprehensive analysis showed that chromosome of P.mirabilis isolates mainly carries multidrug-resistance complex elements harboring resistance to carbapenems genes blaOXA-1, blaNDM-1 and blaTEM-1. Class I integron is the insertion hotspot of IS26, it can be inserted into type I integron at different sites, thus forming a variety of multiple drug resistance decision sites. At the same time, Tn21, Tn7, SXT / R391 Mobile elements cause widespread spread of this drug resistance genes. In conclusion, P.mirabilis isolates from wildlife showed higher resistance to commonly used clinic drugs comparing to those from human. Therefore, wild animals carrying multidrug resistance (MDR) clinical isolates should be paid attention to by the public health. This article is protected by copyright. All rights reserved.

H1N1 influenza is a kind of acute respiratory infectious disease that has a high socioeconomic and medical burden each year around the world. In the past decades, H1N1 influenza viruses have exhibited high resistance to adamantanes, which has become a serious issue. To understand the up-to-date distribution and evolution of H1N1 influenza viruses with adamantanes-resistant mutations, we conducted a deep analysis of 15875 M2 protein and 8351 MP nucleotides sequences. Results of the distribution analyses showed that 77.32% of H1N1 influenza viruses harbored-resistance mutations of which 73.52% were S31N, And the mutant variants mainly appeared in North America and Europe and H1N1 influenza viruses with S31N mutation became the circulating strains since 2009 all over the world. In addition, 80.65% of human H1N1 influenza viruses and 74.61% of swine H1N1 influenza viruses exhibited adamantanes resistance, while the frequency was only 1.86% in avian H1N1 influenza viruses. Studies from evolutionary analyses indicated that the avian-origin swine H1N1 influenza viruses replaced the classical human H1N1 influenza viruses and became the circulating strains after 2009; The interspecies transmission among avian, swine, and human strains over the past 20 years contributed to the 2009 swine influenza pandemic. Results of our study clearly clarify the historical drug resistance level of H1N1 influenza viruses around the world and demonstrated the evolution of adamantanes-resistant mutations in H1N1 influenza viruses. Our findings emphasize the necessity for monitoring the adamantanes susceptibility of H1N1 influenza viruses and draw attention to analyses of the evolution of drug-resistant H1N1 influenza variants.

A localized positive charge on IgG (referred to as a "charge patch") shows an adverse effect on pharmacokinetics (PK), so it would seem to be best practice to avoid charge patches during the discovery stage and closely monitor charge interactions during the development process. In certain circumstances, however, charge patches are required for target binding, in which case completely removing charge patches is not feasible. Therefore, quantitative measurement of a charge patch and its impact on PK is critical to the success of therapeutic antibody development. In this article, we generated mutations of a recombinant human antibody (referred to as mAb1) with disrupted charge patches to investigate how charge patches on IgG antibodies impact both target-binding affinity and PK-related factors. We conclude that it is important to modulate the size of the charge patch in order to balance target-binding affinity and PK.

We previously reported that blue eggshell color in chickens is associated with a partial endogenous retroviral (EAV-HP) insertion in the promoter region of the solute carrier organic anion transporter family member 1B3 (SLCO1B3) gene. The EAV-HP sequence includes numerous regulatory elements, which may modulate the expression of adjacent genes. To determine whether this insertion influences the expression of neighboring genes, we screened the expression of solute carrier organic anion transporter family members 1C1, 1B1 (SLCO1C1, SLCO1B1), and SLCO1B3 in 13 and 10 tissues from female and male Yimeng chickens, respectively. We observed that the insertion only significantly modulated the expression of SLCO1B3 and did not majorly affect that of SLCO1C1 and SLCO1B1. High expression of SLCO1B3 was detected in the shell gland, magnum, isthmus, and vagina of the oviduct in female blue-eggshell chickens. We also observed ectopic expression of SLCO1B3 in the testes of male chickens. SLCO1B3 is typically highly expressed in the liver; however, the EAV-HP insertion significantly reduces SLCO1B3 expression. As a liver-specific transporter, a reduction in the expression of SLCO1B3 may affect liver metabolism, particularly that of bile acids. We also detected higher ectopic expression of SLCO1B3 in the lungs of birds heterozygous for the EAV-HP insertion than in homozygous genotypes. In conclusion, we confirmed that the EAV-HP insertion modifies SLCO1B3 expression, and showed, for the first time, similar expression profile of this gene in all parts of the oviduct in females and testis in males. We also observed different levels of SLCO1B3 expression in the liver, which were associated with the EAV-HP insertion, and significantly higher expression in the lungs of birds with heterozygous genotype. The effects of these changes in the SLCO1B3 expression pattern on the function of the tissues warrant further investigation.

Microorganisms plays an important role in the growth of Pyropia haitanensis. To understand the structural and functional diversity of the microorganism community of P. haitanensis (PH40), the associated metabolic pathway network in cluster of orthologous groups (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG), and carbohydrate-active enzymes (CAZymes) were explored in metagenomic analysis. DNA extraction from gametophytes of P. haitanensis was performed first, followed by library construction, sequencing, preprocessing of sequencing data, taxonomy assignment, gene prediction, and functional annotation. The results show that the predominant microorganisms of P. haitanensis were bacteria (98.98%), and the phylum with the highest abundance was Proteobacteria (54.64%), followed by Bacteroidetes (37.92%). Erythrobacter (3.98%) and Hyunsoonleella jejuensis (1.56%) were the genera and species with the highest abundance of bacteria, respectively. The COG annotation demonstrated that genes associated with microbial metabolism was the predominant category. The results of metabolic pathway annotation show that the ABC transport system and two-component system were the main pathways in the microbial community. Plant growth hormone biosynthesis pathway and multi-vitamin biosynthesis functional units (modules) were the other important pathways. The CAZyme annotation revealed that the starch might be an important carbon source for microorganisms. Glycosyl transferase family 2 (GT2) and glycosyl transferase family 3 (GT3) were the highly abundant families in glucoside transferase superfamily. Six metagenome-assembled genomes containing enzymes involved in the biosynthesis of cobalamin (vitamin B12) and indole-3-acetic acid were obtained by binning method. They were confirmed to belong to Rhodobacterales and Rhizobiales, respectively. Our findings provide comprehensive insights into the microorganism community of Pyropia.

Congenital heart disease (CHD) is the most common birth defect worldwide. Long non-coding RNAs (lncRNAs) have been implicated in many diseases. However, their involvement in CHD is not well understood. This study aimed to investigate the role of dysregulated lncRNAs in CHD. We used Gene Expression Omnibus data mining, bioinformatics analysis, and analysis of clinical tissue samples and observed that the novel lncRNA SAP30-2:1 with unknown function was significantly downregulated in damaged cardiac tissues from patients with CHD. Knockdown of lncRNA SAP30-2:1 inhibited the proliferation of human embryonic kidney and AC16 cells and decreased the expression of heart and neural crest derivatives expressed 2 (HAND2). Moreover, lncRNA SAP30-2:1 was associated with HAND2 by RNA immunoprecipitation. Overall, these results suggest that lncRNA SAP30-2:1 may be involved in heart development through affecting cell proliferation via targeting HAND2 and may thus represent a novel therapeutic target for CHD.

Purpose Cervical squamous cell carcinoma (CSCC) seriously affects women's health worldwide, and it is of great significance to illuminate the specific role of circRNAs in CSCC. Materials and methods Three mRNA datasets, two miRNA datasets and one circRNA dataset of CSCC, downloaded from GEO, were utilized in this study. Differentially expressed mRNAs (DEmRNAs), miRNAs (DEmiRNAs) and circRNAs (DEcircRNAs) were identified, and a ceRNA (DEcircRNA-DEmiRNA-DEmRNA) regulatory network was constructed. GO and pathway analyses of DEcircRNAs and DEmRNAs in the ceRNA regulatory network were performed. Quantitative real-time polymerase chain reaction (qRT-PCR) validation of the expression of the selected DEmRNAs, DEmiRNAs and DEcircRNAs was performed. Results A total of 1356 DEmRNAs, 13 DEmiRNAs and 77 DEcircRNAs were obtained. The ceRNA network contained 3 circRNA-miRNA pairs and 158 miRNA-mRNA pairs, including 3 circRNAs, 3 miRNAs, and 138 mRNAs. Functional annotation of DEmRNAs in the ceRNA regulatory network revealed that these DEmRNAs were significantly enriched in cell cycle, p53 signalling pathway and DNA replication. The qRT-PCR results were generally consistent with those of our integrated analysis. Conclusion In conclusion, we speculate that the regulation of the hsa_circ_0000069/hsa-miR-125b-5p/CDKN2A and hsa_circ_0020594/hsa-let-7c-5p/CCNB2 axes may be involved in CSCC.

This study systematically investigated the phytotoxicity of chicken manure co-composted with tobacco powder and mushroom substrate on seed germination during active phase of composting. All compost products met the sanitation requirements specified in the Chinese national standard; however, only the mushroom substrate compost satisfied the maturity standard. From day 28, the composting entered the end of active phase and the concentrations of K+, Zn2+, Na+, Cu2+ and Fe3+ decreased gradually. Redundancy analysis indicated that the germination index, catalase and peroxidase activities was positively correlated with K+, Zn2+, Na+, Cu2+, Fe3+ and NO3--N, and negatively correlated with NH4+-N, Mg2+ and Ca2+, among which the most significant ions were Fe3+, Mg2+ and Zn2+ for all treatments. The malondialdehyde concentration of germinated seeds had adverse correlation with the above ions parameters.

The irrigating winter wheat one or two times each season was practiced in Huanghuai Winter Wheat Region in order to reduce the over-exploitation of groundwater. In this study, plant height (PH) and its relationship with yield were analyzed in a field study conducted over three growing seasons with 102 wheat varieties irrigated zero to two times at different growth stages. Here, we found that irrigation at the jointing stage increased wheat PH by 6.60–9.70%. Whereas irrigation at the early milking stage had almost no effect on it. Against the wildtype alleles Rht-B1a and Rht-D1a, the genetic background of Rht-B1b and Rht-D1b significantly reduced PH by 14.40–15.50% and 16.90–19.10%, respectively. The combined Rht-B1b and Rht-D1b can reduce PH by 29.10–32.50%. PH was strongly, negatively correlated with grain yield in jointing irrigation (W1) or jointing + milking irrigation (W2) over three growing seasons. In contrast, a significant positive correlation was detected in no-irrigation (W0) in the 2012/2013 wheat growing season, yet no correlation was found in 2013/2014 and 2014/2015. PH was positively correlated with drought coefficient (DRC) in W1 and W2. These results augment our understanding of the relationship between PH and grain yield in different irrigation regimes and provide PH traits for wheat improvement on water saving.

Raffinose and its precursor galactinol accumulate in plant leaves during abiotic stress. RAFFINOSE SYNTHASE (RAFS) catalyzes raffinose formation by transferring a galactosyl group of galactinol to sucrose. However, whether RAFS contributes to plant drought tolerance and, if so, by what mechanism remains unclear. In this study, we report that expression of RAFS from maize (or corn, Zea mays) (ZmRAFS) is induced by drought, heat, cold, and salinity stresses. We found that zmrafs mutant maize plants completely lack raffinose and hyper-accumulate galactinol and are more sensitive to drought stress than the corresponding null-segregant (NS) plants. This indicated that ZmRAFS and its product raffinose contribute to plant drought tolerance. ZmRAFS overexpression in Arabidopsis enhanced drought stress tolerance by increasing myo-inositol levels via ZmRAFS-mediated galactinol hydrolysis in the leaves due to sucrose insufficiency in leaf cells and also enhanced raffinose synthesis in the seeds. Supplementation of sucrose to detached leaves converted ZmRAFS from hydrolyzing galactinol to synthesizing raffinose. Taken together, we demonstrate that ZmRAFS enhances plant drought tolerance through either raffinose synthesis or galactinol hydrolysis, depending on sucrose availability in plant cells. These results provide new avenues to improve plant drought stress tolerance through manipulation of the raffinose anabolic pathway.

LL-37, also called cathelicidin, is an important part of the human immune system, which can resist various pathogens. A plethora of experiments have demonstrated that it has the multifunctional effects of immune regulation, in addition to antimicrobial activity. Recently, there have been increasing interest in its immune function. It was found that LL-37 can have two distinct functions in different tissues and different microenvironments. Thus, it is necessary to investigate LL-37 immune functions from the two sides of the same coin. On the one side, LL-37 promotes inflammation and immune response and exerts its anti-infective and antitumor effects; on the other side, it has the ability to inhibit inflammation and promote carcinogenesis. This review presents a brief summary of its expression, structure, and immunomodulatory effects as well as brief discussions on the role of this small peptide as a key factor in the development and treatment of various inflammation-related diseases and cancers.

The main causes of failure of orthopedic implants are infection and poor bone ingrowth. Surface modification of the implants to allow for long-term antibacterial and osteogenic functions is an effective solution to prevent failure of the implants. We developed silver-rich TiN/Ag nano-multilayers on the surface of titanium alloy with different doses of Ag+ . The antibacterial stability and osteogenesis of the silver-rich surface were determined by evaluating the adhesion and proliferation of Staphylococcus epidermidis, and the adhesion, proliferation, alkaline phosphatase activity, extracellular matrix mineralization, and the expression level of genes involved in osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs). The results demonstrated that the antibacterial rates (Ra) of 5 × 1016 -Ag, 1 × 1017 -Ag, 5 × 1017 -Ag, and 1 × 1018 -Ag were respectively 46.21%, 85.66%, 94.99%, 98.48%, and 99.99%. After subcutaneous implantation in rats or immersion in phosphate buffered saline for up to 12 weeks, the silver-rich surface of the titanium alloy showed long-term stable inhibition of Staphylococcus epidermidis. Furthermore, in vitro and in vivo studies indicated that the Ag-implanted titanium did not show apparent cytotoxicity and that lower Ag+ implanted groups (5 × 1016 -Ag, 1 × 1017 -Ag) had better viability and biological safety when compared with higher Ag+ implanted groups. In addition, when compared with the Ti6Al4V-group, all Ag-implanted groups exhibited enhanced osteogenic indicators in rat BMSCs. Regarding osteogenic indicators, the surfaces of the 5 × 1017 -Ag group had better osteogenic effects than those of other groups. Therefore, the proper dose of Ag+ implanted TiN/Ag nano-multilayers may be one of the options for the hard tissue replacement materials with antibacterial activity and osteogenic functions.

Tetralogy of Fallot (TOF) is the most common complex congenital heart disease (CHD) with uncertain cause. Although long non‐coding RNAs (lncRNAs) have been implicated in heart development and several CHDs, their role in TOF is not well understood. This study aimed to investigate how dysregulated lncRNAs contribute to TOF. Using Gene Expression Omnibus data mining, bioinformatics analysis and clinical heart tissue sample detecting, we identified a novel antisense lncRNA TBX5‐AS1:2 with unknown function that was significantly down‐regulated in injured cardiac tissues from TOF patients. LncRNA TBX5‐AS1:2 was mainly located in the nucleus of the human embryonic kidney 293 (HEK293T) cells and formed an RNA‐RNA double‐stranded structure in the overlapping region with its sense mRNA T‐box transcription factor 5 (TBX5), which is an important regulator in heart development. Knock‐down of lncRNA TBX5‐AS1:2 via promoter hypermethylation reduced TBX5 expression at both the mRNA and protein levels by affecting its mRNA stability through RNA‐RNA interaction. Moreover, lncRNA TBX5‐AS1:2 knock‐down inhibited the proliferation of HEK293T cells. In conclusion, these results indicated that lncRNA TBX5‐AS1:2 may be involved in TOF by affecting cell proliferation by targeting TBX5.

Drought stress is a major threat to maize ( Zea mays L.) yield. Deep-seeding tolerant maize variety can absorb water in deep soil and thus have strong drought tolerance. Therefore, it is of great theoretical and practical importance to study the genetic mechanism of deep-seeding tolerance. In our previous work, we identified a major QTL qMES20-10 controlling maize deep-seeding tolerance on chromosome 10 using an F2:3 population derived from a deep-seeding tolerant inbred line 3681-4 and a common inbred line X178. In this study, a BC3F3:4 population was constructed through background and foreground selection using X178 as the recurrent parent. The major QTL, qMES20-10 , was firstly verified in this BC3F3:4 population. Furthermore, advanced backcross population was constructed through marker-assisted selection, and qMES20-10 was fine-mapped within the interval of 133.3–136.0 Mb on chromosome 10. Moreover, RNA-Seq analysis of two near-isogenic lines screened from the BC3F3:4 families identified the differentially expressed genes, mainly involved in chemical stimulus response, oxidation reduction, and oxidative stress response. These results lay a foundation of further cloning the major QTL qMES20-10.

How egg temperature sets sex In many reptiles, sex is determined by nest temperature during egg incubation. Temperature regulates the expression of an epigenetic modifier gene called Kdm6b , which is responsible for testis development. However, the molecular connection between temperature and sex-specific expression of this factor was previously unknown. Weber et al. have identified a link between temperature and the activation of a key regulator of Kdm6b called signal transducer and activator of transcription 3 (STAT3). After an influx of Ca 2+ at the warmer, female-producing temperature, STAT3 is phosphorylated and silences Kdm6b transcription to repress testis development. Science , this issue p. 303