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Decreased functional β-cell mass is the hallmark of diabetes, but the cause of this metabolic defect remains elusive. Here, we show that the expression levels of the growth factor receptor-bound protein 10 (GRB10), a negative regulator of insulin and mTORC1 signaling, are markedly induced in islets of diabetic mice and high glucose-treated insulinoma cell line INS-1cells. β-cell-specific knockout of Grb10 in mice increased β-cell mass and improved β-cell function. Grb10-deficient β-cells exhibit enhanced mTORC1 signaling and reduced β-cell dedifferentiation, which could be blocked by rapamycin. On the contrary, Grb10 overexpression induced β-cell dedifferentiation in MIN6 cells. Our study identifies GRB10 as a critical regulator of β-cell dedifferentiation and β-cell mass, which exerts its effect by inhibiting mTORC1 signaling.

MicroRNAs (miRNAs) act as regulators of plant development and multiple stress responses. Here we demonstrate that the rice miR171b-SCL6-IIs module regulates the balance between blast resistance, grain yield, and flowering. miR171b-overexpressing rice plants (OX171b) displayed increased rice blast resistance accompanied with enhanced defense responses and late heading, whereas blocking miR171b expression in rice (MIM171) led to greater susceptibility to blast disease, associated with compromised defense responses and early heading. Either overexpressing or silencing of miR171b significantly affected plant height and number of filled seeds per panicle (seed-setting rate), resulting in decreased grain yield. miR171b targets SCL6-IIa, SCL6-IIb, and SCL6-IIc, whose expression was suppressed in OX171b but increased in MIM171. Mutants of SCL6-IIa, SCL6-IIb, and SCL6-IIc all displayed phenotypes like that of OX171b, including markedly increased blast disease resistance, slightly decreased grain yield, and delayed flowering. Amounts of miR171b increased gradually in leaves during the vegetative stage but decreased gradually in panicles during the reproductive stage, whereas SCL6-IIs displayed the reverse expression pattern. Together, these results suggest that the expression of miR171b was time- and space-dependent during the rice growth period and regulated the balance between rice blast disease resistance, grain yield, and flowering via SCL6-IIs, and that appropriate accumulation of miR171b is essential for rice development.

The avoidance of detection by predators and parasites is critical to survival. Two complex mechanisms for such avoidance are mimicry and camouflage, with fossils providing valuable insight into the evolution of these strategies. Such fossil evidence is, however, rare, and the frequent partial and lopsided occurrence. Here, we report a diverse insect assemblage exhibiting these adaptations from mid-Cretaceous Kachin amber (99 million years ago), including plant mimesis in Tridactylidae (pygmy mole crickets) and debris-carrying camouflage in Gelastocoridae (toad bugs) and Psocodea (bark lice). Critically, Mesozoic plant mimesis in Tridactylidae is supported by our Siamese Network analysis, a Deep Learning model and potentially powerful tool for investigating ancient mimicry. Together with previously known records, our fossils demonstrate that most extant debris-carrying insects (eight groups with direct camouflage) had evolved exogenous camouflage by the mid-Cretaceous. Our results suggest that a complex biological response was already widespread among insects in mid-Cretaceous ecosystems during the rise of angiosperms, probably in response to similar selective pressures as experienced by their extant counterparts.

Objective Small cell lung cancer (SCLC) transformation had previously been reported mainly in epidermal growth factor receptor (EGFR) mutant adenocarcinoma. However, the underlying genomic profile remains unclear. Our study aim to find the evolution and genotypic characteristic of SCLC transformation. Patients and methods Thirty-one SCLC transformation patients who were initially diagnosed as Non-Small Cell Lung Cancer (NSCLC) patients were included. Whole exome sequencing (WES) of both primary and transformed re-biopsy lesions was conducted on 12 patients. Clinical characteristics were analyzed using R software (v.3.6.1). Results Our study included 31 patients, of whom, three had lung squamous cell carcinoma. Six patients did not carry EGFR mutations. Thirty patients received chemotherapy for SCLCs. The disease control rate (DCR) was 96.7%, and the median progression-free survival (PFS) was 4.03 months. The median time to transformation was 33.07 months, and the median overall survival (OS) was 62.08 months. Somatic mutation analysis showed that besides TP53, RB1, and EGFR, there was a high occurrence of mutations to CSMD3 and ADAMTS19, especially in the EGFR-wild type (EGFR-wt) group. Concerning mutational signature, the EGFR-mutant (EFFR-mut) transformed group favored an APOBEC (apolipoprotein B mRNA editing catalytic polypeptide-like)-associated mutation pattern (P = 0.16). DNA Damage Repair (DDR)-related signatures were significantly enriched in the EGFR-wt transformed group (P = 0.034). Additionally, clonal evolution analysis revealed that all patients had the same main trunk genes in the phylogenetic tree. Transformed SCLCs are not sensitive to immunotherapy, possibly due to increased tumor heterogeneity. Conclusion Our results indicated that the EGFR-wt patients could also transform to SCLCs, but they have different genetic features with EGFR-mut patients. SCLC-transformed patients respond to classical chemotherapy and have a better prognosis than those with classical SCLCs.

This study was conducted to explore the correlations between the expression, methylation, and various clinicopathological factors of purinergic P2X1 receptor (P2RX1) and the prognosis of patients with gastrointestinal tumors. The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) databases were used to analyze the expression of P2RX1 in different types of gastrointestinal cancers. Kaplan-Meier analysis and univariate Cox regression analysis were used to analyze the correlations between P2RX1 expression and the prognosis of various gastrointestinal tumors. Correlations between P2RX1 expression and N6 methyladenine (m6A)-related genes as well as immune checkpoint genes were analyzed by R packages (R version: 4.0.3) based on TCGA database. The association between P2RX1 methylation level and the prognosis of patients with gastrointestinal cancers was analyzed using the MethSurv database. In order to explore the biological functions of P2RX1 in hepatocellular carcinoma, the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis were carried out using R software. In order to evaluate the correlations between P2RX1 and tumor immune infiltration, Spearman correlation test was performed. The correlations between P2RX1 expression and immune score as well as immune checkpoint genes were analyzed based on TCGA and Tumor Immune Estimation Resource (TIMER) databases. The expression of P2RX1 was found to be significantly downregulated in gastrointestinal tumors except in cholangiocarcinoma (P < 0.05). High expression of P2RX1 tended to present better prognosis in hepatocellular carcinoma (P < 0.05). It was noted that cg06475633 of P2RX1 presented a higher methylation level compared with other CpG sites in hepatocellular carcinoma. Overall, six CpGs of P2RX1 were associated with significant prognosis in patients with hepatocellular carcinoma (P < 0.05). Among all the 20 m6A-related genes, Wilms’ tumor 1-associating protein (WTAP) was the most strongly correlated with P2RX1 in hepatocellular carcinoma. Gene enrichment analysis showed that P2RX1 is widely involved in the proliferation, activation, organization, and differentiation of various immune cells. After investigating the TIMER database, P2RX1 was found to be tightly correlated with immune infiltrating cells in gastrointestinal tumors, especially with dendritic cells. Moreover, P2RX1 was found to be strongly positively associated with programmed cell death 1 (PD1), programmed death-ligand 1 (PD-L1), and cytotoxic T-lymphocyte-associated protein 4 (CTLA4) in hepatocellular carcinoma (P < 0.05). In conclusion, the dual role of P2RX1 in cancers and its involvement in the recruitment as well as regulation of tumor infiltrating cells in gastrointestinal cancers may be appreciated through this study.

Heterosis refers to the superior phenotypes observed in hybrids. Cytoplasmic male sterility (CMS) system plays an important role in cotton heterosis utilization. However, the global gene expression patterns of CMS-D2 and its interaction with the restorer gene Rf1 remain unclear. Here, the full-length transcript sequencing was performed in anthers of the CMS-D2 restorer line using PacBio single-molecule real-time sequencing technology. Combining PacBio SMRT long-read isoforms and Illumina RNA-seq data, 107,066 isoforms from 44,338 loci were obtained, including 10,086 novel isoforms of novel genes and 66,419 new isoforms of known genes. Totally 56,572 alternative splicing (AS) events, 1146 lncRNAs, 61 fusion transcripts and 10,466 genes exhibited alternative polyadenylation (APA), and 60,995 novel isoforms with predicted open reading frames (ORFs) were further identified. Furthermore, the specifically expressed genes in restorer line were selected and confirmed by qRT-PCR. These findings provide a basis for upland cotton genome annotation and transcriptome research, and will help to reveal the molecular mechanism of interaction between Rf1 and CMS-D2 cytoplasm.

Progressive familial intrahepatic cholestasis type 1 (PFIC1) results from biallelic pathogenic variants in ATP8B1. This study sought second pathogenic variants in ATP8B1 by whole-genome sequencing (WGS) in four unrelated low γ-glutamyl transpeptidase cholestasis patients in whom clinical suspicion of PFIC1 was high and gene-panel or Sanger sequencing had identified only one pathogenic variant in ATP8B1. Sanger sequencing confirmed WGS findings and determined the origin of each variant. Novel nonrecurrent structural variants in three patients (patient 1 to patient 3) were identified in trans: g.55396652_55403080del (6427-bp deletion), g.55335906_55346620dup (10,715-bp duplication), and g.55362063_55364293dup (2231-bp duplication). One synonymous variant in patient 4 was recognized in trans (c.1029GA, p. Thr343Thr) and demonstrated as deleterious. In conclusion, WGS improves genetic diagnostic yield in PFIC1. These findings expand the gene-variant spectrum associated with familiar intrahepatic cholestasis 1 (FIC1) disease and for the first time report tandem duplication in ATP8B1 associated with cholestasis.

Phlebiopsis is a genus of poroid crust fungi belong to the family Phanerochaetaceae. The genus contains 19 species, which show a widespread distribution. In this paper, a strain of Phlebiopsis sp., named WS01, was isolated from tubers of Gastrodia elata collected in southern China. The endophytic fungus WS01 bearing fruiting-body was cultivated successfully in the fruiting body media. The characterized strain, in hyphae of the hymenium with colorless walls and cystidia lacking revealed that it morphologically differs from other Phlebiopsis species. A combined dataset of the internal transcribed spacer (ITS), the large subunit regions of nuclear ribosomal RNA gene (LSU) and RNA polymerase II largest subunit gene (rpb1) was used to confirm the phylogenetic placement of this species, and phylogenetic analyses were carried out with maximum likelihood and bayesian inference methods. The results provide evidence that the species represents a new member of the Phlebiopsis clade in the family Phanerochaetaceae, in which the new species formed a monophyletic lineage with a strong support (100% ML, 1.00 BPP) and then grouped with P. pilatii. According to a signature sequence for a new species, the strain is, therefore, proposed as Phlebiopsis xuefengensis sp. nov. It is the first record of Phlebiopsis fungi from the plant endophytes and the first novel fungal species isolated from Gastrodia elata.

KEY MESSAGE: qGSN5, a novel quantitative trait locus coordinating grain size and grain number in rice, was fine-mapped to an 85.60-kb region. GS3 may be a suppressor of qGSN5. Grain size and grain number are two factors that directly determine rice grain yield; however, the underlying genetic mechanisms are complicated and remain largely unclear. In this study, a chromosome segment substitution line (CSSL), CSSL28, which showed increased grain size and decreased grain number per panicle, was identified in a set of CSSLs derived from a cross between 93-11 (recipient) and Nipponbare (donor). Four substitution segments were identified in CSSL28, and the substitution segment located on chromosome 5 was responsible for the phenotypes of CSSL28. Thus, we defined this quantitative trait locus (QTL) as grain size and grain number 5 (qGSN5). Cytological and quantitative PCR analysis showed that qGSN5 regulates the development of the spikelet hull by affecting cell proliferation. Genetic analysis showed that qGSN5 is a semi-dominant locus regulating grain size and grain number. Through map-based cloning and overlapping substitution segment analysis, qGSN5 was finally delimited to an 85.60-kb region. Based on sequence and quantitative PCR analysis, Os05g47510, which encodes a P-type pentatricopeptide repeat protein, is the most likely candidate gene for qGSN5. Pyramiding analysis showed that the effect of qGSN5 was significantly lower in the presence of a functional GS3 gene, indicating that GS3 may be a suppressor of qGSN5. In addition, we found that qGSN5 could improve the grain shape of hybrid rice. Together, our results lay the foundation for cloning a novel QTL coordinating grain size and grain number in rice and provide a good genetic material for long-grain hybrid rice breeding.

Cancer metastasis accounts for nearly 90% of all cancer deaths. Metastatic cancer progression requires both cancer cell migration to the site of the metastasis and subsequent proliferation after colonization. However, it has long been recognized that cancer cell migration and proliferation can be uncoupled; but the mechanism underlying this paradox is not well understood. Here we report that TNFAIP8 (tumor necrosis factor-α-induced protein 8), a “professional” transfer protein of phosphoinositide second messengers, promotes cancer cell migration or metastasis but inhibits its proliferation or cancer growth. TNFAIP8-deficient mice developed larger tumors, but TNFAIP8-deficient tumor cells completely lost their ability to migrate toward chemoattractants and were defective in colonizing lung tissues as compared to wild-type counterparts. Mechanistically, TNFAIP8 served as a cellular “pilot” of tumor cell migration by locally amplifying PI3K–AKT and Rac signals on the cell membrane facing chemoattractant; at the same time, TNFAIP8 also acted as a global inhibitor of tumor cell growth and proliferation by regulating Hippo signaling pathway. These findings help explain the migration–proliferation paradox of cancer cells that characterizes many cancers.

Fatty acid uptake is extremely important for the survival and growth of the intracellular parasite Toxoplasma gondii. In this study, CRISPR-Cas9 gene editing technology was used to investigate the role of four lipid synthesis enzymes, namely, glycerol-3-phosphate dehydrogenase (G3PDH), malonyl CoA-acyl carrier protein transacylase (FabD), acyl-ACP thiolesterase (TE), and diacylglycerol acyltransferase (DGAT), in the virulence and infectivity of Type I RH and Type II Prugniaud (Pru) strains of T. gondii. Immunofluorescence analysis of the tachyzoite stage showed that FabD protein was located in the apicoplast; however, the expression level of the other three proteins was undetectable. Compared with wild-type (WT) strains, the growth of RHΔG3PDH, RHΔTE, and RHΔDGAT in vitro and their virulence in vivo were not significantly different. However, RHΔFabD exhibited a significantly reduced growth rate, compared with the WT strain. The deletion of FabD attenuated the virulence of Type II Pru strain and reduced the formation of cysts in vivo. These data improved our understanding of the role of lipid synthesis enzymes in the pathogenesis of T. gondii.

Charophyte assemblages from the Cretaceous/Paleogene (K/Pg) boundary transition in the JZK-2 borehole of the Jiaolai Basin (eastern China) were studied. The charophyte flora includes Tolypella grambastii, Peckichara praecursoria, Microchara cristata, Microchara prolixa, Chara changzhouensis, Lamprothamnium ellipticum, Nodosochara (Turbochara) specialis, Lychnothamnus aff. vectensis, and Lychnothamnus lanpingensis n. comb. The discovery of the ostracod species Porpocypris sphaeroidalis Guan in the upper section of the JZK-2 borehole strongly indicates the arrival of the Paleocene. Three floras of the latest Cretaceous to the early Paleocene were compared among the Jiaolai, Pingyi, and Songliao basins. The Pingyi Basin is special because it is the only carbonate palaeolake that occurred throughout the K/Pg transition in China. It is characterized by the appearance of the Cretaceous brackish water element, Feistiella anluensis (Wang), and is dominated by the Paleocene species, Peckichara varians Grambast. The Songliao and Jiaolai floras grow in clastic lakes; the Jiaolai Basin is thought to correspond to an altitude lake, which was formed in the coastal mountains with high palaeoelevation (≥ 2.0 km), while the Songliao Basin is presumed to have been formed in a low elevation site according to previous studies. This difference in altitude explains why the Songliao Basin is more species-rich than the Jiaolai Basin.

Neonatal hypoxic-ischaemic (HI) injury is a serious complication of neonatal asphyxia and the leading cause of neonatal acute death and chronic neurological injury, and the effective therapeutic method is lacking to improve patients' outcomes. We reported in this study that panax notoginseng saponin (PNS) may provide a treatment option for HI. HI model was established using neonatal Sprague-Dawley rats and then intraperitoneally injected with different dosage of PNS, once a day for 7 days. Histological staining and behavioural evaluations were performed to elucidate the pathological changes and neurobehavioural variation after PNS treatment. We found PNS administration significantly reduced the infarct volume of brain tissues and improved the autonomous activities of neonatal rats, especially with higher dosage. PNS treatment at 40 mg/kg reduced neuronal damage, suppressed neuronal apoptosis and depressed astroglial reactive response. Moreover, the long-term cognitive and motor functions were also improved after PNS treatment at 40 mg/kg. Importantly, PNS treatment elevated the levels of BDNF and TrkB but decreased the expression of p75NTR both in the cortex and hippocampus of HI rats. The therapeutic efficacy of PNS might be correlated with PNS-activated BDNF/TrkB signalling and inactivation of p75NTR expression, providing a novel potential therapy for alleviating HI injury.

Lily (Lilium spp.), with its beautiful flower, is an important horticultural crop and a popular ornamental plant, but because the abundant pollen pollutes the flowers and surroundings, its use is restricted. To solve this problem, the mechanism of pollen development in lily needs to be analyzed. However, the complex and delicate process of anther development in lily remains largely unknown. In this study, LoUDT1, a bHLH transcription factor (TF), was isolated and identified in lily. LoUDT1 was closely related to OsUDT1 of Oryza sativa and AtDYT1 of Arabidopsis. It was localized in the cytoplasm and nucleus and showed no transcriptional activation in yeast cells. LoUDT1 interacted with another bHLH TF, LoAMS, and the interaction depended on their BIF domains. LoUDT1 and LoAMS were both expressed in the anthers but showed different expression patterns. LoUDT1 was continuously expressed during the entire development of anthers, whereas LoAMS was only highly expressed early in anther development. With overexpression of LoUDT1 in Arabidopsis, normal anther development was affected and defective pollens were produced, which caused partial male sterility of transgenic plants. These defects depended on the level of LoUDT1 accumulation. By contrast, with the appropriate expression of LoUDT1 in a dyt1-3 mutant, normal pollen grains were produced, showing partial fertility. Thus, LoUDT1 might be a key regulator of anther development in lily. By further increasing the understanding of anther development, the results of this study can provide a theoretical basis for the molecular breeding of pollen-free lilies.

The role of B7‐H3 in acute myeloid leukemia (AML) is not fully understood. Two previous studies investigating its expression and significances in AML are partially different. In this study, we aimed to systematically characterize the genomic and immune landscape in AML patients with altered B7‐H3 expression using multi‐omics data in the public domain. We found significantly increased B7‐H3 expression in AML compared to either other hematological malignancies or healthy controls. Clinically, high B7‐H3 expression was associated with old age, TP53 mutations, wild‐type WT1 and CEBPA, and the M3 and M5 FAB subtypes. Moreover, we observed that increased B7‐H3 expression correlated significantly with a poor outcome of AML patients in four independent datasets. Gene set enrichment analysis (GSEA) revealed the enrichment of the “EMT” oncogenic gene signatures in high B7‐H3 expressers. Further investigation suggested that B7‐H3 was more likely to be associated with immune‐suppressive cells (macrophages, neutrophils, dendritic cells, and Th17 cells). B7‐H3 was also positively associated with a number of checkpoint genes, such as VISTA (B7‐H5), CD80 (B7‐1), CD86 (B7‐2), and CD70. In summary, we uncovered distinct genomic and immunologic features associated with B7‐H3 expression in AML. This may lead to a better understanding of the molecular mechanisms underlying B7‐H3 dysregulation in AML and to the development of novel therapeutic strategies., In this paper, we found significantly increased B7‐H3 expression in AML compared to either other hematological malignancies or healthy controls. Importantly, we observed that increased B7‐H3 expression correlated significantly with a poor outcome of AML patients in four independent datasets. We also uncovered distinct genomic and immunologic features associated with B7‐H3 expression in AML.

Biological technology has presented a promising application for phosphine (PH3) purification; however, the degradation characteristics and microbial community of PH3 biopurification systems have ra...

MsCML46 enhances tolerance to abiotic stresses through alleviating osmotic stress and oxidative damage by regulating the expression of stress-related genes to optimize osmolytes levels and antioxidant enzyme activity in transgenic tobacco. Abiotic stresses are major environmental factors that constraint crop productivity worldwide. Various stimuli regulate intracellular calcium levels and calcium-mediated signal transduction, and cellular responses. Ca2+ signals are perceived by different Ca2+ receptors. Calmodulin-like protein (CML) is one of the best-characterized Ca2+ sensors which shares sequence similarity with highly conserved calmodulin (CaM) ubiquitously expressed in plants. Currently, the molecular and physiological functions of CMLs are largely unknown. In this study, the MsCML46 was characterized in alfalfa (Medicago sativa cv. Zhaodong) under freezing stress. Results showed that MsCML46 was localized to the cytoplasm of Arabidopsis, and its expression was strongly elevated by cold, drought, salt, saline-alkali, and ABA treatments. Overexpressing MsCML46 in tobacco enhanced tolerance to freezing, drought, and salt stresses as evidenced by improved contents of osmotic regulatory solutes and antioxidant enzyme activity but decreased reactive oxygen species (ROS) accumulation. Furthermore, cold, drought, and salt stresses increased the expression of stress-related genes in transgenic tobacco. MsCML46 binds free Ca2+ to promote signal transduction and maintain higher K+/Na+ ratio. In this way, it protects intracellular homeostasis under sodium ion toxicity. These results suggest that MsCML46 plays a crucial role in resisting abiotic stresses and can be exploited in genetic engineering for crops.

Salicylic acid (SA) acts antagonistically to jasmonic acid (JA) in plant immunity. We previously reported that CATALASE2 (CAT2) promotes JA‐biosynthetic acyl‐CoA oxidase (ACX) activity to enhance plant resistance to necrotrophic Botrytis cinerea, and SA represses JA biosynthesis through inhibiting CAT2 activity, while the underlying mechanism remains to be further elucidated. Here, we report that the truncated CAT2 N‐terminus (CAT2‐N) interacts with and promotes ACX2/3, and CAT2‐N‐overexpressing plants have increased JA accumulation and enhanced resistance to B. cinerea B05.10, but compromised antagonism of SA on JA. Catalase inhibitor treatment or mutating CAT2 active amino acids abolished CAT2 H2O2‐decomposing activity but did not affect its promotion of ACX2/3 activity via interaction. CAT2‐N, a truncated protein with no catalase activity, interacted with and promoted ACX2/3. Overexpressing CAT2‐N in Arabidopsis plants resulted in increased ACX activity, higher JA accumulation, and stronger resistance to B. cinerea B05.10 infection. Additionally, SA dramatically repressed JA biosynthesis and resistance to B. cinerea in the wild type but not in the CAT2‐N‐overexpressing plants. Together, our study reveals that CAT2‐N can be utilized as an accelerator for JA biosynthesis during plant resistance to B. cinerea B05.10, and this truncated protein partly relieves SA repression of JA biosynthesis in plant defence responses., N‐terminal fragment of CATALASE2 interacts with and promotes acyl‐CoA oxidases implicated in JA biosynthesis and enhances plant resistance to Botrytis cinerea, thus CATALASE2‐N could be a genetic tool conferring plant resistance.

Fat mass and obesity-associated protein (FTO), an N6-methyladenosine (m6A) demethylase, participates in tumor progression and metastasis in many malignancies, but its role in colorectal cancer (CRC) is still unclear. Here, we found that FTO protein levels, but not RNA levels, were downregulated in CRC tissues. Reduced FTO protein expression was correlated with a high recurrence rate and poor prognosis in resectable CRC patients. Moreover, we demonstrated that hypoxia restrained FTO protein expression, mainly due to an increase in ubiquitin-mediated protein degradation. The serine/threonine kinase receptor associated protein (STRAP) might served as the E3 ligase and K216 was the major ubiquitination site responsible for hypoxia-induced FTO degradation. FTO inhibited CRC metastasis both in vitro and in vivo. Mechanistically, FTO exerted a tumor suppressive role by inhibiting metastasis-associated protein 1 (MTA1) expression in an m6A-dependent manner. Methylated MTA1 transcripts were recognized by an m6A “reader”, insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2), which then stabilized its mRNA. Together, our findings highlight the critical role of FTO in CRC metastasis and reveal a novel epigenetic mechanism by which the hypoxic tumor microenvironment promotes CRC metastasis.

In this study, the effect of coelomic fluid isolates (CFIs) from sea cucumbers (Stichopus japonicus) on autolysis of minced muscle tissue of S. japonicus was investigated. The coelomic fluid of S. japonicus at the concentration of 90% showed suppression effects on activities of papain and trypsin with inhibitory rate of 26.6% and 40.4%, respectively. Moreover, the effect of CFIs on autolysis of minced muscle tissue from S. japonicus was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and trichloroacetic acid (TCA)-soluble oligopeptide assay. CFIs exhibited significantly inhibitory effect on autolytic muscle mince samples with inhibitory rate at 56.2–87.2%. Both heat and proteinase treatments did not influence the inhibitory effect of CFIs on autolysis of minced muscle tissue from S. japonicus. These results suggest that the non-protein and thermally stable substances contribute to the inhibitory effect of CFIs, which can be used to improve sea cucumber quality.

A novel pathogenic strain Vibrio 20190611023 was isolated from the hepatopancreas of moribund cultured Penaeus vannamei suffering from black gill disease. This strain was identified as V. brasiliensis based on the phylogenetic analyses of 16S rDNA gene and five other housekeeping genes (i.e., gapA, ftsZ, mreB, topA and gyrB). Some biochemical features of this strain were determined with an API 20NE system, and its haemolytic activity was determined using a sheep blood agar plate. The pathogenicity of this isolate 20190611023 was confirmed by the experimental challenge tests and histopathological examinations. P. vannamei were challenged via reverse gavage with different doses of bacterial suspensions. The calculated median lethal dose (LD50 ) was (3.16 ± 1.78) × 105 CFU/g (body weight). Moreover, antibiotic susceptibility tests were performed, the results of which showed that the strain 20190611023 was sensitive to chloramphenicol, compound sulphamethoxazole, ciprofloxacin, doxycycline and oxacillin, but resistant to erythromycin, kanamycin, gentamicin, cefoperazone, ceftriaxone, cefamezin and piperacillin. To our knowledge, this is the first report for demonstrating V. brasiliensis as a shrimp pathogen, which expands the host range of V. brasiliensis infection. The present study highlights that more attention should be paid to this novel pathogen in intensive shrimp aquaculture.

The sterile triploid crucian carp (TC) has growth superiority which is generated from crossing between diploid red crucian carp (Carassius carassius red var. ♀) (RC) and tetraploid hybrid linage [derived from RC (♀) × common carp (Cyprinus carpio, ♂) (CC)]. Hypothalamus is an important control center for hormone releasing participating in gonadal development. In the present study, the microRNA (miRNA) alternations in the hypothalamus from RC, CC and TC were analyzed. A total of 143 known miRNAs and 519 novel miRNAs were identified from the hypothalamus. Thereinto, miR-7a, miR-128 and miR-9-3p were the most abundant miRNAs accounting for 13.37%, 12.01% and 11.15% of all the miRNA reads, respectively. A total of 109, 208 and 228 differentially expressed miRNAs (DEMs) were identified from RC vs TC, CC vs TC and CC vs RC, respectively. For these DEMs, 46 miRNA-mRNA pairs were identified by miRNA target prediction. In addition, using weighted gene co-expression network analysis (WGCNA), 17 modules of co-expressed miRNAs and mRNA were constructed and yellow module had the highest correlation with the sterile triploid. A total of 107 novel miRNAs, 29 conserved miRNAs and 1343 genes were contained in yellow module and metabolic processes, biosynthetic processes, chromosome organization, cytoskeleton organization, gene expression and axis specification were enriched GO terms in this module. The top three hub miRNAs were let-7a, miR-100 and miR-122 which were correlated with 1086, 1172 and 1222 genes, respectively. These results indicated that the hub miRNAs and their regulated gene network play a central role in controlling endocrine secretion and gamete maturation. In addition, the abnormal expression of these miRNAs contributes to the sterility of TC. The present evidence provides insights into the miRNA and gene constructed network of sterility in triploid fish.

Ligustici Radix (Chinese name: maoqianhu) consists of the dried roots of Ligusticum brachylobum Franch., which is mainly distributed in the Yunnan and Sichuan provinces. This herbal medicine has been primarily used for the treatment of cough in traditional Chinese medicine. Ligustici Radix is rich in coumarin derivatives. Interestingly, enantiomers and diastereomers are widely used for these coumarins, thus posing a great challenge for in-depth chemical profile characterization. In the present study, a new analytical platform, achiral-chiral liquid chromatography-tandem mass spectrometry (achiral-chiral LC-MS/MS) was configured to profile the chemical composition of Ligustici Radix. Because achiral and chiral columns were serially coupled, especially enantiomers, both chemically and enantiomerically selective separations could be accomplished simultaneously. The newly configured achiral-chiral LC-MS/MS platform did not require any electronic valve; hence, it could overcome the drawbacks of heart-cutting achiral-chiral two-dimensional LC, i. e., sophisticated instrumentation and limited reproducibility due to the use of electronic valve(s) and the undesired retention time shift across different analytical runs. Some available candidates for chemically selective or enantiomerically selective separation were assayed; then, Capcell core RP-C18 column that was packed with core-shell type particles, and AD-RH column embedding amylose coated particles were employed the achiral and the chiral columns, respectively. The narrow-bore core-shell RP-C18 column served as the front tool to achieve efficient chemoselective separation of coumarin analogs, and enantioselective enantiomers were obtained by using a wide-bore AD-RH chiral column. The predictive multiple reaction monitoring (predictive MRM) mode allowed for the sensitive detection of potential components, and an enhanced product ion (EPI) scan, which was a unique function of Qtrap-MS, was programmed to record the MS2 spectra for all captured signals and thus aid structural annotation. Online energy-resolved mass spectrometry (online ER-MS) was introduced to pursue the suitable collision energy for each compound; in particular, inferior collision energy instead of the optimal one was utilized to suppress the response of the primary components such as praeruptorin A, B and pteryxin. The criteria to judge enantiomers or not included identical quantitative and qualitative precursor-to-product ion transitions, identical quantitative versus qualitative responses, and longer retention times from achiral-chiral LC over single-column achiral LC. As a result, a total of sixty components were observed and structurally identified. In particular, enantiomerically selective separations were achieved for eight enantiomers, cis-khellactone (CKL), qianhucoumarin G (QC-G), pteryxin (Pte), praeruptorin A (PA), cis-3'-isovaleryl-4'-acetylkhellactone (IAK), praeruptorin B (PB), praeruptorin E (PE), and cis-3',4'-diisovalerylkhellactone (DIK). Notably, none of the enantiomers were present as racemates; instead, the proportion of one enantiomer in each pair was greater than the other. Achiral-chiral LC-predictive MRM is a feasible choice for the quantitative and qualitative analyses of Ligustici Radix as well as other herbal medicines characterized by enantiomers and diastereomers.

As a signaling molecule, hydrogen sulfide (H2S) plays an indispensable role in the modulation of ripening and senescence in fruits and vegetables. To explore the role of H2S in regulating metabolism of postharvest tomato, ripening-related physiological parameters, activities of antioxidant enzymes and gene expression were analyzed in H2S-fumigated tomato fruits. These results show that H2S significantly delayed the color transition and softening of tomato fruit, and maintained higher level of flavonoids and lower level of anthocyanin during storage. Besides, H2S could maintain higher level of nutritional-related metabolites, such as reducing sugar, ascorbic acid during postharvest storage. Moreover, H2S decreased the rate of O2− production, inhibited the production of H2O2 and malondialdehyde (MDA), enhanced the activities of antioxidant enzymes including ascorbate peroxidase (APX), superoxide dismutase (SOD), catalase (CAT) and guaiacol peroxidase (POD) in tomato fruits, while reduced the activities of phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO) and lipoxygenase (LOX). Besides, the expression of the antioxidant-encoding genes SlCAT2, SlPOD12 was generally upregulated with H2S fumigation. Principal component analysis (PCA) suggests that H2S induced significant discrepancy mainly to the differences in firmness, anthocyanin, flavonoid and the activity of guaiacol peroxidase (POD), and the correlation analysis further shows that H2S affected pigment metabolism and nutritional quality. In conclusion, H2S could maintain better appearance and nutritional quality, and prolong the storage period of postharvest tomato fruits through activating the antioxidative system.

Penicillium digitatum is the primary pathogen that causes serious yield losses worldwide. In our previous study, CsWRKY transcription factors (TFs) and some genes associated with immunity were identified in citrus fruits after P. digitatum infection, but little information is available in the literature on the mechanisms of TFs in citrus disease resistance. In this study, the possible mechanisms of CsWRKY65 participating in the establishment of disease resistance were investigated. Results show that CsWRKY65 was a transcriptional activator in the nucleus. The dual-luciferase transient assays and electrophoretic mobility shift assays showed that CsWRKY65 bound with CsRbohB, CsRbohD, CsCDPK33, and CsPR10 promoters to activate gene transcription. Besides, the transient overexpression of CsWRKY65 induced reactive oxygen species accumulation and increased PR gene expression in Nicotiana benthamiana leaves. The transient overexpression of CsWRKY65 in the citrus peel enhanced the disease resistance against P. digitatum. In conclusion, CsWRKY65 is likely to be involved in regulating the disease resistance to P. digitatum of citrus fruits by directly activating the expressions of CsRbohB, CsRbohD, CsCDPK33, and CsPR10. This study provides new information for the mechanism of citrus WRKY TFs participating in the establishment of disease resistance.

MYCNOS is an oncogenic lncRNA in liver cancer, but its role in glioblastoma (GBM) is unknown. We predicted that MYCNOS might interact with miR-216b, which targets FOXM1 to perform tumor suppressive roles. This study was performed to analyze the role of MYCNOS in GBM and explore its potential interactions with miR-216b and FOXM1. MYCNOS expression in paired GBM and non-tumor tissues from 62 GBM patients was analyzed by RT-qPCR. The interaction between MYCNOS and miR-216b was predicted by IntaRNA 2.0 and confirmed by dual luciferase activity assay. Overexpression of MYCNOS, miR-216b, and FOXM1 was achieved in GBM cells, followed by performing RT-qPCR and Western blot to explore the relationship among them. CCK-8 assay was performed to explore the role of MYCNOS, miR-216b, and FOXM1 in regulating GBM cell proliferation. MYCNOS was upregulated in GBM tissues compared to the paired non-tumor tissues. MYCNOS is predicted to interact with miR-216b, but overexpression of MYCNOS and miR-216b failed to affect each other's expression significantly. Dual luciferase activity assay showed that MYCNOS and miR-216b could directly interact with each other. MYCNOS overexpression increased the expression of FOXM1, which is a direct target of miR-216b. Cell proliferation assay showed that MYCNOS and FOXM1 overexpression resulted in an increased proliferation rate of GBM cells, while miR-216b overexpression suppressed cell proliferation. Moreover, MYCNOS overexpression suppressed the role of miR-216b. MYCNOS may regulate FOXM1 expression of by serving as an endogenous sponge of miR-216b axis to promote the proliferation of GBM cells.

Maize (Zea mays L.) germplasm in China Summer maize ecological region (CSM) or central corn-belt of China is diverse but has not been systematically characterized at molecular level. In this study, genetic variation, genome diversity, linkage disequilibrium patterns, population structure, and characteristics of different heterotic groups were studied using 525,141 SNPs obtained by Genotyping-By-Sequencing (GBS) for 490 inbred lines collected from researchers at CSM region. The SNP density is lower near centromere, but higher near telomere region of maize chromosome, the degree of linkage disequilibrium (r2) vary at different chromosome regions. Majority of the inbred lines (66.05%) show pairwise relative kinship near zero, indicating a large genetic diversity in the CSM breeding germplasm. Using 4849 tagSNPs derived from 3618 haplotype blocks, the 490 inbred lines were delineated into 3 supergroups, 6 groups, and 10 subgroups using ADMIXTURE software. A procedure of assigning inbred lines into heterotic groups using genomic data and tag-SNPs was developed and validated. Genome differentiation among different subgroups measured by Fst, and the genetic diversity within each subgroup measured by GD are both large. The share of heterotic groups that have significant North American germplasm contribution: P, SS, IDT, and X, accounts about 54% of the CSM breeding germplasm collection and has increased significantly in the last two decades. Two predominant types of heterotic pattern in CSM region are: M-Reid group × TSPT group, and X subgroup × Local subgroups.

The forkhead box O3a protein (FoxO3a) has been reported to regulate tumour invasion and migration, but little is known about the molecular mechanism or its role in trophoblast invasion and migration into the uterus. In this study, we aim to explore its role in trophoblast development and placenta‐related pregnancy complications and the potential mechanism. Levels of FoxO3a and its phosphorylated form (p‐FoxO3a) in placental tissue from healthy pregnant women and pre‐eclampsia patients were first compared. Then, HTR‐8/SVneo cells were transfected with lentiviral vectors to deplete and overexpress FoxO3a. Western blot, immunohistochemistry, Cell Counting Kit‐8, wound‐healing assay, Matrigel invasion assay, cell apoptosis, cell cycle assay, RNA sequencing, qRT‐PCR and ChIP‐qPCR were performed on the cells to study the potential role of FoxO3a and the underlying mechanism. We found the expression of FoxO3a was decreased, whereas p‐FoxO3a was increased in pre‐eclampsia placentae. FoxO3a depletion significantly reduced transcription of the promoter region of intercellular cell adhesion molecule‐1 (ICAM1) gene in ChIP assays and led to reduced invasion and migration of trophoblast cells, arrested cell cycle in G1 phase and increased apoptosis under oxidative stress. Our results suggested that FoxO3a may play a role in the regulation of trophoblast invasion and migration during placental development, which may be because of its affinity to the ICAM1 promotor.

If age boundaries are arbitrarily or roughly defined, age-related analyses can result in questionable findings. Here, we aimed to delineate the uniquely age-dependent immune features of coronavirus disease 2019 (COVID-19) in a retrospective study of 447 patients, stratified according to age distributions of COVID-19 morbidity statistics into well-defined age-cohorts (2-25y, 26-38y, 39-57y, 58-68y, and 69-79y). Age-dependent susceptibilities and severities of the disease were observed in COVID-19 patients. A comparison of the lymphocyte counts among the five age-groups indicated that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection led to age-dependent lymphopenia. Among the lymphocyte subsets, the CD8+ T cell count alone was significantly and age-dependently decreased (520, 385, 320, 172, and 139 n/μl in the five age-groups, respectively). In contrast, the CD4+ T cell, B cell, and natural killer cell counts did not differ among age-cohorts. Age and CD8+ T cell counts (r=‒0.435, p

Cephalodiones A-D (1-4), the first example of C19 -norditerpenoid dimers, were isolated and fully characterized from a Cephalotaxus plant. These new skeletal natural products shared a unique tricyclo[6.4.1.12,7 ]tetradeca-3,5,9,11-tetraene-13,14-dione core that was capped in both ends with rigid multicyclic ring systems either C2 -symmetrically or asymmetrically. Compounds 1-4 were proposed to be biosynthetically produced by the [6+6]-cycloaddition of two identical C19 -norditerpenoid troponoids, which was validated by the semisyntheses of dimers 2-4. Moreover, some compounds showed significant inhibition on Th17 cell differentiation.

The discovery of effective therapeutic treatments for cancer via cell differentiation instead of antiproliferation remains a great challenge. Cyclin-dependent kinase 2 (CDK2) inactivation, which overcomes the differentiation arrest of acute myeloid leukemia (AML) cells, may be a promising method for AML treatment. However, there is no available selective CDK2 inhibitor. More importantly, the inhibition of only the enzymatic function of CDK2 would be insufficient to promote notable AML differentiation. To further validate the role and druggability of CDK2 involved in AML differentiation, a suitable chemical tool is needed. Therefore, we developed first-in-class CDK2-targeted proteolysis-targeting chimeras (PROTACs), which promoted rapid and potent CDK2 degradation in different cell lines without comparable degradation of other targets, and induced remarkable differentiation of AML cell lines and primary patient cells. These data clearly demonstrated the practicality and importance of PROTACs as alternative tools for verifying CDK2 protein functions.

β-N-acetylhexosaminidases have gained much attention for synthesis of functional oligosaccharides, but the relatively low conversion ratio limited their industrial applications. A β-N-acetylhexosaminidase gene (CmHex187) from Catenibacterium mitsuokai was successfully expressed in Escherichia coli. The recombinant enzyme (CmHex187) was purified, biochemically characterized and subjected to the synthesis of human milk oligosaccharides (HMOs). CmHex187 displayed optimal pH and temperature of 5.5 and 45 °C, respectively. The enzyme showed strong transglycosylation activity, as it efficiently catalyzed the synthesis of an important component of HMO lacto-N-triose II (LNT2) using 0.015 M p-nitrophenyl N-acetyl-β- d -glucosaminide and 1.2 M lactose at pH 7.0 and 60 °C for 1.5 h, with a high conversion ratio of 44.3 %. The formed LNT2 promoted the growth of tested Lactobacillus and Bifidobacteria strains. The excellent enzymatic properties and high LNT2 synthesis ability of CmHex187 may make it a good candidate in LNT2 production.

Background The microRNA (miRNA) miR-133a-1 has been identified as a tumor suppressor in breast cancer. However, the underlying mechanisms of miR-133a-1 in breast cancer have not been fully elucidated. This study aimed to explore the targets of miR-133a-1 in breast cancer using an integrated bioinformatics approach. Methods Human SKBR3 breast cancer cells were transfected with miR-133a-1 or a miRNA negative control (miRNA-NC) for 48 hours. The RNA-seq sequencing technique was performed to identify the differential expression of genes induced by miR-133a-1 overexpression. Functional enrichment analysis was conducted to determine the target genes and pathways involved in breast cancer. Results Breast cancer patients with high levels of miR-133a-1 expression commonly showed longer overall survival compared to patients with a low level of miR-133a-1 expression. Using Cuffdiff, we identified 1,216 differentially expressed genes induced by miR-133a-1 overexpression, including 653 upregulated and 563 downregulated genes. MOCS3 was the most upregulated gene and KRT14 was the most downregulated gene. The top 10 pathways related to the differentially expressed genes were identified through Gene Ontology (GO) enrichment analysis. Sex-determining region Y-box 9 (SOX9) demonstrated the highest semantic similarities among the differentially expressed genes. Since SOX9 and CD44 were hub nodes in the protein-protein interaction network, the SOX9 gene may be a target of miR-133a-1 in breast cancer. Conclusions This report provides useful insights for understanding the underlying mechanisms in the pathogenesis of breast cancer.

Background High-risk human papilloma virus (hrHPV) is the main causal factor of cervical precancer and cancer when persistent infection is left untreated. Previous studies have confirmed the vaginal microbiota is associated with HPV infection and the development of cervical lesions. The microbiota at different parts of the female genital tract is closely related but different from each other. To analyze the distinction between the vaginal and cervical microbiota of hrHPV(+) women in China, one hundred subjects were recruited, including 10 patients with HPV16/18(+) and cervical carcinoma, 38 patients with HPV16/18(+) but no cervical carcinoma, 32 patients with other hrHPV(+) and 20 healthy controls with HPV(−). Vaginal and cervical microbiota were separately tested through next-generation sequencing (NGS) targeting the variable region (V3-V4) of the bacterial ribosome 16S rRNA gene. Results HrHPV(+) subjects had higher percentages of vaginal douching history (P = 0.001), showed more frequent usage of sanitary pads (P = 0.007), had more sex partners (P = 0.047), were more sexually active (P = 0.025) and more diversed in ways of contraception (P = 0.001). The alpha diversity of the cervical microbiota was higher than that of the vagina. The cervical microbiota consisted of a lower percentage of Firmicutes and a higher percentage of Proteobacteria than the vagina at the phylum level. Sphingomonas, belonging to α-Proteobacteria, was almost below the detection limit in the vagina but accounted for five to 10 % of the bacteria in the hrHPV(−) cervix (PPPseudomonas, belonging to γ-Proteobacteria, could hardly be seen in the normal vagina and shared a small percentage in the normal cervix but was significantly higher in the HPV16/18(+) (PPGardnerella, Prevotella, Atopobium and Sneathia, between the cevix and vigina. Conclusions The proportion of Proteobacteria was significantly higher in the cervical microbiota than that of vagina. The hrHPV infection and cervical cancer was positively associated with Pseudomonas and negatively associated with Sphingomonas. It is of great improtance to deeply explore the cervical microbiota and its function in cervical cacinogenesis.

Aeromonas veronii can cause a variety of diseases such as sepsis in humans and animals. However, there has been no effective way to eradicate A. veronii. In this study, the intracellular antibacterial activities of the C-terminal aminated marine peptide N6 (N6NH2) and its D-enantiomer (DN6NH2) against A. veronii were investigated in macrophages and in mice, respectively. The result showed that DN6NH2 with the minimum inhibitory concentration (MIC) of 1.62 μM is more resistant to cathepsin B than N6NH2 (3.23 μM). The penetration percentages of the cells treated with 4–200 μg/mL fluorescein isothiocyanate (FITC)-DN6NH2 were 52.5–99.6%, higher than those of FITC-N6NH2 (27.0–99.1%). Both N6NH2 and DN6NH2 entered macrophages by macropinocytosis and an energy-dependent manner. DN6NH2 reduced intracellular A. veronii by 34.57%, superior to N6NH2 (19.52%). After treatment with 100 μg/mL DN6NH2, the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β were reduced by 53.45%, 58.54%, and 44.62%, respectively, lower than those of N6NH2 (15.65%, 12.88%, and 14.10%, respectively); DN6NH2 increased the IL-10 level (42.94%), higher than N6NH2 (7.67%). In the mice peritonitis model, 5 μmol/kg DN6NH2 reduced intracellular A. veronii colonization by 73.22%, which was superior to N6NH2 (32.45%) or ciprofloxacin (45.67%). This suggests that DN6NH2 may be used as the candidate for treating intracellular multidrug-resistant (MDR) A. veronii. • DN6NH 2 improved intracellular antibacterial activity against MDR A. veronii. • DN6NH 2 entered macrophages by micropinocytosis and enhanced the internalization rates. • DN6NH 2 effectively protected the mice from infection with A. veronii.

Light is the most important environmental cue signaling the transition from skotomorphogenesis to photomorphogenesis, thus affecting plant development and metabolic activity. How the light response mechanisms of maize seedlings respond to fluctuations in the light environment has not been well characterized to date. In this study, we built a gene coexpression network from a dynamic transcriptomic map of maize seedlings exposed to different light environments. Coexpression analysis identified ten modules and multiple genes that closely correlate with photosynthesis and characterized hub genes associated with regulatory networks, duplication events, domestication and improvement. In addition, we identified that 38% of hub genes underwent duplication events, 74% of which are related to photosynthesis. Moreover, we captured the dynamic expression atlas of gene sets involved in the chloroplast photosynthetic apparatus and photosynthetic carbon assimilation in different light environments, which should help to elucidate the key mechanisms and regulatory networks that underlie photosynthesis in maize. Insights from this study provide a valuable resource to better understand the genetic mechanisms of the response to fluctuations in the light environment in maize.

With the increase in pet keeping among families in China, there has been a corresponding increase in the incidence of allergic diseases such as bronchial asthma and allergic rhinoconjunctivitis. Pet allergies pose important social and health problems, making their management in children crucial. There is an increasing trend of sensitization to cat and dog dander among Chinese children. The process underlying allergic sensitization is complex and is associated with an IgE-mediated type I hypersensitivity reaction that involves the upregulation of type 2 T-helper (Th2) cells, resulting in Th2 immunity. Compared with traditional serum-specific IgE (sIgE) and skin tests, molecular diagnosis may be better at predicting the progression of diseases. The association between pet keeping and allergic diseases in children remains controversial. Although the hygiene hypothesis is widely accepted in Chinese children, exposure to pets in early life remains a risk factor for allergies. Pet avoidance and allergy immunotherapy (AIT) are recommended for children with pet allergies. However, pet avoidance is difficult to maintain, and AIT is not widely available in China. There is an imbalance between the increase in allergic diseases and the shortage of prevention and treatment options. We hope that advancements in medicine, diagnosis, and healthcare policies will help resolve this issue.

Phytoliths are microscopic particles of amorphous silica produced by plants that have vital importance for environmental change and carbon sequestration. This study investigated phytoliths and their morphotype assemblages, and phytolith-occluded carbon (PhytOC) along a marsh degradation gradient to clarify the significance of phytoliths for evaluating carbon sequestration dynamics during marsh degradation on the Zoige Plateau, China. Soil samples from non-degraded marsh (NDM), lightly degraded marsh (LDM), moderately degraded marsh (MDM), and seriously degraded marsh (SDM) were collected. Soil phytolith shape and PhytOC were determined using heavy liquid suspension, microscope observations, and alkali-solution spectrophotometry. The dominant phytolith morphotypes among the 15 identified in the soils from the differently degraded marshes were lanceolate, rectangular, and elongate, which originated from Poaceae and Cyperaceae. Soil phytolith morphotype assemblages show potential for studying past vegetation and climatic succession in alpine wetlands. The soil phytolith and PhytOC content exhibited no significant differences between the NDM and LDM samples. Compared with the NDM soil, phytolith content was significantly lower in the MDM (33.8–58.8%) and SDM (52.7–65.4%) soils, respectively, and PhytOC content was also significantly lower (by 31.0–57.1% and 47.5–74.4%, respectively). The soil phytolith and PhytOC content decreased significantly with soil depth, especially once the marsh has become moderately degraded. Alpine marsh degradation reduced the accumulation of soil PhytOC by decreasing the PhytOC input flux, thereby altering the long-term carbon sequestration mechanism of phytoliths in this alpine-marsh wetland ecosystem.

Verticillium wilt is a vascular disease caused by Verticillium dahliae, which has a serious impact on the yield and quality of eggplant. MiRNAs regulates plant growth, development, environmental adaptation and other life activities by controlling the expression of target genes. Although miRNAs have been systematically studied by high-throughput sequencing in eggplant (Solanum melongena L.), their target genes are not identified systematically, which would provide the basis for exploring the mechanism of miRNA mediated gene regulation in response to Verticillium wilt of eggplant. In this study, degradome sequencing was performed using two libraries separately prepared from mock-inoculated and V. dahliae-inoculated seedlings of eggplants. A total of 122 target genes for 49 miRNAs families were identified. Of them, 62 target genes were significantly induced by V. dahliae, including 34 targets were found only in the inoculated seedlings library and 28 targets were shown to be differentially expressed between both two libraries. Further analysis showed that a conserved miRNA could target various genes ranging from one up to 15 or a target gene could be cleaved by 1–5 different miRNAs. QRT-PCR analysis on five miRNAs and their target genes confirmed the reliability of the degradome sequencing data. GO analysis revealed that target genes owned diverse biological functions and transcription regulation was the largest functional category. Analysis on KEGG showed that target genes were mainly involved in plant-pathogen interaction pathway and plant hormone signal transduction pathway to resist disease. This result will provide basic information for further studying the regulatory mechanism of eggplant miRNAs in response to Verticillium wilt infection.

Hundreds of insect genome sequences have been reported; however, most sequencing projects have not determined the sex chromosomes, and no Y chromosomes from a heterometabolous insect have been identified and characterized to date. The brown planthopper (Nilaparvata lugens Stål) is the most economically damaging pest to rice and is also an ideal research subject for paddy ecology and functional genomics. We previously assembled a draft female genome mainly using second-generation sequencing technologies, with a contig N50 of only 24 kb, due to the large size and excessive repetitive regions in the N. lugens genome. Here, we utilize third-generation sequencing technologies and Hi-C data to generate a high-quality male N. lugens assembly with a contig N50 of 1.01 Mb, a scaffold N50 of 69.96 Mb and more than 95.6% of the assembled bases located on 16 chromosomes. Fourteen autosomes and two sex chromosomes (X + Y) were identified, filling in the gap related to the Y chromosome in heterometabolous insects. A total of 18,021 protein-coding genes and 6423 long-noncoding RNAs were predicted with full-length cDNA sequencing data. All 315 of the Y chromosome genes (Y-genes) were derived from autosomal and X-chromosome duplications. Large-scale RNA interference (RNAi) experiments were conducted against the N. lugens Y-genes, demonstrating that 7 Y-genes were essential for normal BPH development or male organ development, suggesting the importance of Y-genes. The first identified Y chromosome in heterometabolous insects will help gain more insight into sex determination, fertility and chromosome evolution.

Four unprecedented macrocyclic nonapeptides, orberryamides A–D (1–4), were isolated from Glycosmis pentaphylla (orangeberry) and structurally characterized by obtaining solid data from numerous ana...

The newly identified Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has resulted in a global health emergency because of its rapid spread and high mortality. The molecular mechanism of interaction between host and viral genomic RNA is yet unclear. We demonstrate herein that SARS-CoV-2 genomic RNA, as well as the negative-sense RNA, is dynamically N6-methyladenosine (m6A)-modified in human and monkey cells. Combined RIP-seq and miCLIP analyses identified a total of 8 m6A sites at single-base resolution in the genome. Especially, epidemic strains with mutations at these identified m6A sites have emerged worldwide, and formed a unique cluster in the US as indicated by phylogenetic analysis. Further functional experiments showed that m6A methylation negatively regulates SARS-CoV-2 infection. SARS-CoV-2 infection also triggered a global increase in host m6A methylome, exhibiting altered localization and motifs of m6A methylation in mRNAs. Altogether, our results identify m6A as a dynamic epitranscriptomic mark mediating the virus–host interaction.

Insulin resistance (IR) is a complex pathological condition resulting from the dysregulation of cellular response to insulin hormone in insulin-dependent cells and is recognized as a pathogenic hallmark and strong risk factor for metabolic syndrome. The present study aims to elucidate the molecular mechanism of the pathogenesis of IR. Here, we used human umbilical vein endothelial cells (HUVECs) to establish the IR cell model induced by 1 × 10(−6) mmol/L insulin. After 48 h, reactive oxygen species (ROS) and glucose consumption were measured by DCFH-DA and GOD-POD methods, respectively. The results of Microarray analysis demonstrated that there were 10 differentially expressed miRNAs (DEMs) selected based on Fold change (FC) and P value in the IR cell model compared with HUVECs. The enriched gene ontology (GO) terms analysis showed that the target genes of these 10 DEMs were significantly enriched in biological process, cellular component and molecular function, and the significantly enriched Kyoto Encyclopedia of Genes or Genomes (KEGG) pathways mainly include AMPK signaling pathway and PI3K signaling pathway. Amongst all, the expression level of miR-3138 was highest in the IR cell model evaluated by qRT-PCR. Through Targetscan, KSR2 mRNA was predicted as a target of miR-3138. And mRNA and protein expression levels of miR-3138, KSR2, GLUT4, AMPK, PI3K, Akt were examined using qRT-PCR and Western blotting, respectively. The interaction between miR-3138 and KSR2 was evaluated by dual-luciferase reporter assay. Our results showed that miR-3138 significantly deteriorated the IR of HUVECs via KSR2/AMPK/GLUT4 signaling pathway.

Two new benzo[de]isoquinoline derivatives, 4-phenyl-benzo[de]isoquinoline-1,3-dione (1) and 4-(4-hydroxyphenyl)-benzo[de]isoquinoline-1,3-dione (2), were isolated from 70% ethanol extract of the rh...

MicroRNA-205 (miR-205) is believed to be related to the progress of tumors. HOXD9 has been proved to be expressed abnormally in several kinds of cancers. However, the role of miR-205 and HOXD9 in breast cancer remains unclear. The biological role of miR-205 in breast cancer cell proliferation and chemoresistance was investigated. The expression of miR-205 in clinical tissues and breast cancer cell lines were analyzed using quantitative real-time PCR test (qRT-PCR). Overexpression and knockdown models of miR-205 were established to study cell proliferation and chemotherapy-resistant. Moreover, the potential relationships between miR-205 and HOXD9/Snail1 were measured using qRT-PCR, western blot, and chemotherapy-resistant study. miR-205 was lowly expressed in breast cancer tissues and cell lines. Overexpression of miR-205 could inhibit cell proliferation and chemotherapy-resistance. Moreover, we proved that miR-205 could target the HOXD9-Snail1 axis to suppress triple negative breast cancer cell proliferation and chemoresistance. The activation of Snail1 gene by HOXD9 was also proved in this study. The present study may provide a novel insight for the therapeutic strategies of breast cancer through targeting miR-205/HOXD9/Snail1.

Transforming growth factor (TGF-β) plays an important role in the development of deer antlers. The purpose of this study was to investigate the role of long noncoding RNA in the transcriptional regulation of TGF-β1 and its relationship with the proliferation and differentiation of antler chondrocytes. High-throughput sequencing was used to screen lncRNAs related to TGF-β1. Next, the overexpression plasmid and interference sequence of target lncRNA27785.1 were constructed and transfected into chondrocytes. We found that lncRNA27785.1 inhibited the proliferation and migration of chondrocytes and delayed the transition of cells from G1 to S phase. qRT-PCR and Western blot analysis indicated that the overexpression of lncRNA27785.1 may downregulate mRNA and protein expression of TGF-BR2, Smad3, pSmad3, and Smad4. Our findings highlight lncRNA27785.1 as an inhibitor of chondrocytes proliferation and differentiation by negatively regulating the TGF-β/Smad signaling pathway; this implicates an important regulatory role for long noncoding RNA in the regeneration of antler.

In this study, we reassessed the taxonomic position of Typhlomys (Rodentia: Platacanthomyidae) from Huangshan, Anhui, China, based on morphological and molecular evidence. Results suggested that Typhlomys is comprised of up to six species, including four currently recognized species (Typhlomys cinereus, T. chapensis, T. daloushanensis, and T. nanus), one unconfirmed candidate species, and one new species (Typhlomys huangshanensis sp. nov. ). Morphological analyses further supported the designation of the Huangshan specimens found at mid-elevations in the southern Huangshan Mountains (600 m to 1 200 m a.s.l.) as a new species.

Studies have shown that hyperglycemia aggravates brain damage by affecting vascular endothelial function. However, the precise mechanism remains unclear. Male Sprague-Dawley rat models of diabetes were established by a high-fat diet combined with an intraperitoneal injection of streptozotocin. Rat models of traumatic brain injury were established using the fluid percussion method. Compared with traumatic brain injury rats without diabetic, diabetic rats with traumatic brain injury exhibited more severe brain injury, manifested as increased brain water content and blood-brain barrier permeability, the upregulation of heme oxygenase-1, myeloperoxidase, and Bax, the downregulation of occludin, zona-occludens 1, and Bcl-2 in the penumbra, and reduced modified neurological severity scores. The intraperitoneal injection of a nitric oxide synthase inhibitor N(5)-(1-iminoethyl)-L-ornithine (10 mg/kg) 15 minutes before brain injury aggravated the injury. These findings suggested that nitric oxide synthase plays an important role in the maintenance of cerebral microcirculation, including anti-inflammatory, anti-oxidative stress, and anti-apoptotic activities in diabetic rats with traumatic brain injury. The experimental protocols were approved by the Institutional Animal Care Committee of Harbin Medical University, China (approval No. ky2017-126) on March 6, 2017.