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Lipopolysaccharide-induced TNF-α factor (LITAF) is an important transcription factor mediating the expression of inflammatory cytokines, including TNF-α, which is involved in the innate immune defence against infectious agents. In this study, the Scylla paramamosain LITAF (LITAF) gene was cloned and identified from S. paramamosain using RT-PCR and RACE. The full length cDNA of LITAF was 1116 bp, contained an ORF of 495 bp encoding a polypeptide of 164 amino acids with predicted molecular mass of 17.86 kDa and a theoretical isoelectric point of 8.51. Sequence alignment showed that LITAF possessed a typical LITAF domain with two CXXC motifs and eight conserved cysteine residues. Phylogenetic analysis indicated that LITAF clustered closely with known LITAFs from Eriocheir sinensis and Litopenaeus vannamei. qRT-PCR analysis showed LITAF mRNA transcript expressed ubiquitously in all examined tissues with the highest expression in haemocytes, while lower expression in hepatopancreas. Its expression levels displayed a trend of up-regulation and then returned to its initial value in haemocytes and gill, post challenge with LPS, Poly (I : C), and Vibrio alginolyticus, respectively. Furthermore, the LITAF protein was verified to be located in hyalinocytes, semi-granulocytes and granulocytes, and was distributed throughout the cytoplasm and nucleus, especially in cytoplasm. These results provide a basis for further investigating the roles of LITAF in the immune signaling pathways as a transcription factor in S. paramamosain.

The interactive effect of dietary inactivated Lactobacillus plantarum and berberine on black sea bream, Acanthopagrus schlegelii, was investigated with three diets designated as D1 (Con: basal diet), D2 (LP: basal diet + 400 mg/kg L. plantarum), and D3 (LPBB: basal diet + 400 mg/kg L. plantarum + 50 mg/kg berberine) and fed to juvenile black sea bream ( 5.67 ± 0.05 g) for 56 days. The growth performance and feed utilization parameters, as well as intestinal trypsin activity, were significantly improved in the LP and LPBB groups ( P < 0.05 ). Fish fed the LPBB diet showed better serum and hepatic antioxidant capacity, whilst the LP group had better hepatic antioxidant capacity, than the control fish ( P < 0.05 ). Intestinal IgM and C3 levels significantly increased in the LPBB fish than the rest of the groups ( P < 0.05 ). NF-κB was significantly upregulated in the LP group ( P < 0.05 ). Nrf2 and IL-10 were significantly upregulated, whilst Keap1b and NF-κB were significantly downregulated in the LPBB group ( P < 0.05 ). These findings show that a combination of inactivated L. plantarum and berberine in the diet of black sea bream can improve the immune response and antioxidant capacity than a diet with only the inactivated probiotic, whilst both diets can equally improve growth performance.

Oil-water separation with high efficiency and low energy consumption is a tremendous challenge in the green treatment of oily wastewater. In this paper, a novel filtration method with combined granular media for collaborative removal emulsified oil and suspended solids (SS) was proposed, followed by the exploration of demulsification feasibility and oil removal mechanism. The effect of the operation and structural parameters of the filter bed on oil separation performance was thoroughly investigated, and its feasibility for raw oily wastewater treatment was also explored. A remarkable demulsification performance was observed with the combined granular media filter, and a balance of separation efficiency and pressure drop in the emulsified oily wastewater filtration was also achieved subsequently. Effective oil droplet capture and coalescence were observed with a high speed camera system, and pore clogging could be avoided in combined media. The optimal parameters of the combined media filter (CMF) were concluded to be a combined media ratio of 1:1, a superficial velocity of 0.20 m min

Clostridium autoethanogenum protein (CAP) is a single‐cell protein derived from ethanol production and may have potential to become a substitute for fishmeal in aquafeeds. A 70‐day feeding trial was conducted with black sea bream (Acanthopagrus schlegelii) (mean initial weight 6.03 ± 0.09 g) to evaluate partial replacement of fishmeal with CAP in diets. Seven isonitrogenous and isoenergetic diets were formulated with graded levels of CAP (0, 4.85, 9.70, 14.55, 19.40, 38.80 and 58.20%) to replace fishmeal. The inclusion of CAP at all dietary levels tested did not significantly affect the growth performance (p > .05). Fish fed the CAP58.20% diet showed a significantly lower feeding rate, with significantly higher protein efficiency ratio and feed efficiency ratio compared with fish fed the other diets (p

Purpose: Cells recognize mechanical cues from the extracellular environment through interactions with matrix proteins, such as collagen. Following injury, fibrotic deposition of collagen can lead to aberrant cell differentiation and failed healing. Aligned collagen matrices can drive PDGFRα+ mesenchymal progenitor cells (MPCs) towards osteochondral lineages, leading to heterotopic ossification (HO). Previous work has shown that reducing mechanical strain by immobilizing a limb prevents the formation of an aligned extracellular matrix (ECM), leading to altered MPC differentiation, however, the cellular mechanisms of matrix reorganization following injury remain unclear. Using novel bioinformatics approach, we identified discoidin domain receptor 2 (DDR2) as a key MPC specific tyrosine kinase receptor that interacts with the fibrillar collagen matrix. We hypothesized that DDR2 activation leads to regulation of ECM alignment and therefore serves as a novel upstream regulator of mechanotransductive signaling following musculoskeletal repair. Methods: Heterotopic ossification was induced using a proven mouse model of 30% total body surface area burn with concurrent Achilles’ transection in Ddr2 deletion (Ddr2slie/slie) and littermate control mice on the C57/BL6J background. In separate experiments, ankle joint immobilizers were placed on injured mice for 1, 2 or 3 weeks after burn/tenotomy. Tissue from Ddr2 deleted, immobilized and mobile control mice were harvested for immunofluorescence histology, second harmonic generation of collagen fiber histology at 1, 3 and 9 weeks after injury (SHG) (n=4/group). HO site tissue was also harvested prior to injury and at 1 and 6 weeks after burn/tenotomy and processed for Single-Cell RNA sequencing (scRNA seq) and single nucleus assay for transposase-accessible chromatin (snATAC) using 10X Sequencing and downstream analysis using Seurat 4.0.1 and Signac 3.1.5. Results: Single-cell RNA sequencing showed that genes for collagen type 1 and 3 and Ddr2 are highly upregulated in MPCs following injury. SnATAC showed increased open chromatin reads in the promoter of MPCs following injury (not shown). Utilizing SHG, we found that the collagen matrix is aligned at three-weeks following injury, but one week of limb immobilization was enough to prevent aligned collagen. The active phosphorylated form of DDR2 (pDDR2) was decreased in PDGFRα+ MPCs within immobilized samples, suggesting that DDR2 may affect matrix alignment. Ddr2slie/slie had more disorganized collagen matrix nine weeks following injury and significantly reduced HO. Conclusions: This is the first work to alter collagen matrix organization by genetic modification and mobilization protocols and identifies DDR2 as a critical upstream receptor in matrix organization and aberrant progenitor differentiation following injury. DDR2 antagonism could have therapeutic potential in preventing heterotopic ossification following traumatic injury.

ObjectiveWe previously demonstrated that promoting Beclin-1–dependent autophagy is cardiac protective during endotoxemia shock, suggesting that autophagy-based approaches may become a promising therapeutic strategy for sepsis. In this study, we applied both genetic and pharmacological approaches to evaluate whether Beclin-1 activation improves sepsis outcomes in a model of pneumonia-induced sepsis.MethodsSepsis was induced in mice by Klebsiella pneumoniae infection via intubation, and outcomes of clinical sickness scores, systemic infection, inflammation, survival, and pulmonary pathology were examined. Evaluation of Beclin-1 activation was achieved by comparing strains of C57BL/6J wild type and Becn1F121A that carries a transgenic expression of Beclin-1–active mutant F121A, and by comparing animal groups treated with Beclin-1–activating peptide, Tat-beclin-1 peptide (TB-peptide), or with vehicle control. The status of autophagy in the lung tissue was examined in autophagy reporter mice, CAG-RFP-EGFP-LC3, by fluorescence microscopy.ResultsPulmonary infection by K. pneumoniae produced an insufficient, maladaptive autophagy in the lung. Activation of Beclin-1 by forced expression of active mutant Becn1F121A or by treatment with TB-peptide enhanced autophagy and significantly reduced sickness scores, systemic infection, and circulating and pulmonary cytokine production. Both approaches demonstrated notable benefits in limiting post-infection pathogenesis in the lung, such as decreases in alveolar congestion, hemorrhage, infiltration of inflammatory cells, and alveolar wall thickness.ConclusionData suggest that targeted activation of Beclin-1 alleviates adverse outcomes of pneumonia-induced sepsis, and thus, possess a therapeutic potential.

Silicon-containing heterocyclic compounds are widely used in synthetic organic chemistry, material chemistry and medicinal chemistry due to their unique physicochemical properties. Thus, from the drug and material discovery viewpoints, development of methodology to access diverse silacycles is of great significance and in great demand. Among these, transition metal catalysis provides a powerful and straightforward synthetic method to form diverse silacycles in a step-economical fashion. This article summarized the recent progress in transition metal catalyzed silicon-containing heterocycles synthesis.

Methanol-to-olefin (MTO) is an important non-petroleum chemical process for the preparation of light olefins. However, the MTO process consumes copious amounts of water and produces large amounts of untreated effluent. Therefore, the realization of efficient wastewater treatment and recycling is key to the green low-carbon development of MTO. Here, a cooperative process combining swirl regenerating micro-channel separation (SRMS) and combined fibrous coalescence (CFC) technologies was proposed to separate high contents of oil and suspended matter in MTO wastewater. Using a pilot device with a treatment capacity of 1 m

Pain is a central feature of soft tissue trauma, which under certain contexts, results in aberrant osteochondral differentiation of tissue-specific stem cells. Here, the role of sensory nerve fibers in this abnormal cell fate decision is investigated using a severe extremity injury model in mice. Soft tissue trauma results in NGF (Nerve growth factor) expression, particularly within perivascular cell types. Consequently, NGF-responsive axonal invasion occurs which precedes osteocartilaginous differentiation. Surgical denervation impedes axonal ingrowth, with significant delays in cartilage and bone formation. Likewise, either deletion of Ngf or two complementary methods to inhibit its receptor TrkA (Tropomyosin receptor kinase A) lead to similar delays in axonal invasion and osteochondral differentiation. Mechanistically, single-cell sequencing suggests a shift from TGFβ to FGF signaling activation among pre-chondrogenic cells after denervation. Finally, analysis of human pathologic specimens and databases confirms the relevance of NGF-TrkA signaling in human disease. In sum, NGF-mediated TrkA-expressing axonal ingrowth drives abnormal osteochondral differentiation after soft tissue trauma. NGF-TrkA signaling inhibition may have dual therapeutic use in soft tissue trauma, both as an analgesic and negative regulator of aberrant stem cell differentiation., Soft tissue trauma can result in aberrant osteochondral differentiation of local mesenchymal progenitor cells. Here the authors show that, in mice, soft tissue trauma results in NGF expression by perivascular cells, which leads to axonal invasion and drives abnormal osteochondral differentiation, and show that this process can be prevented by inhibition of NGF signaling.

RecQ-mediated genome instability protein 1 (RMI1) is an important component of the BLM-Topo IIIα-RMI1-RMI2 complex and plays a critical role in maintaining genome stability. However, the cellular functions of RMI1 in response to ionizing radiation (IR) are poorly understood. In this study, we found that

As an important plant single cell model and textile application materials, poorly known about fiber color formation in cotton, which is sensitively regulated by environmental signals. Our studies underline the importance of photo signal on sensitive fiber color formation and characterize fiber color early initiation (15 DPA) and late accumulated metabolites (45 DPA) in different lighting condition. The results revealed 236 differential metabolites between control and shading, of which phenylpropanoids metabolites accounted for 20%, including uncharacterized novel metabolites and pathways. Furthermore, the early initiation specific genes respond to the absence of light are highly correlated with phenylpropanoid metabolites related to pigmentation. The current study reveals the complex pathways involving early initiation regulation and late metabolic pathways. In addition, the collection composed of uncharacterized photoinduced metabolites and early initiation signaling/regulatory genes were identified, which are important resources for understanding fiber color formation. This report provides new insight into molecular regulatory and biochemical basis underlying photoinduced fiber color formation in cotton.

Mitochondria‐associated membranes (MAMs) are essential to mitochondria. This study was to determine whether endotoxemia rearranges MAMs in the heart, and whether Beclin‐1 regulates this process. Wild‐type mice and mice with a cardiac‐specific overexpression of Beclin‐1 (Becn1‐Tg), or a heterozygous knockout of Beclin‐1 (Becn1 +/−) were given lipopolysaccharide (LPS) challenge. In the heart, the ultrastructure of MAMs was examined by electron microscopy and the histology evaluated by immunostaining. Additionally, MAMs were isolated by ultracentrifugation, and their content and function were quantified. The effects of Beclin‐1‐activating peptide (TB‐peptide) on MAMs were also examined. Data showed that endotoxemia decreased both the total mass and the function of MAMs, and these deficiencies became worse in Becn1 +/− mice but were alleviated in Becn1‐Tg and TB‐peptide‐treated mice. Responses of myocardial MAMs to LPS and to TB‐peptide were additionally examined in AC16 human cardiomyocytes. In vitro findings recaptured the effects of LPS and TB‐peptide in cardiomyocytes; the challenge of LPS reduced the level and activity of MAMs, and TB‐peptide attenuated this defect. Together, the results suggest a new function of Beclin‐1 in improving cardiac MAMs during endotoxemia, providing a mechanism for the previously identified role of Beclin‐1 in protection of mitochondria and cardiac function.

The prevention and treatment of coronary heart disease (CHD) is a difficult problem to be solved urgently. Genetic factors play a crucial role in CHD development. This study aimed to investigate the association of GAS5/METTL14/ESR1 polymorphisms with CHD susceptibility. We carried out a case-control study that included 506 patients and 506 healthy subjects to detect the correlation between GAS5/METTL14/ESR1 polymorphisms and CHD risk in a Chinese population. Odds ratios (OR) and 95% confidence intervals (CI) were computed to assess the associations. Our study showed that GAS5 rs17359906 (OR 2.32, p = 0.020) and rs75315904 (OR 0.41, p = 0.039) were related to the risk of CHD in females. ESR1 rs6927072 (OR 1.76, p = 0.007) and rs4870061 (OR 0.74, p = 0.036) correlated with CHD risk in age ≤ 60 years. GAS5 rs17359906 (OR 0.10, p = 0.032) and ESR1 rs3020308 (OR 2.73, p = 0.041) were associated with an increased susceptibility to CHD in smokers. We also found that METTL14 rs4834698 (OR 1.57, p = 0.044) and ESR1 rs4870061 (OR 0.62, p = 0.040) were associated with CHD susceptibility in non-drinkers. Besides, METTL14 rs17050450 (OR 0.48, p = 0.029) and ESR1 rs3853248 (OR 1.61, p = 0.018) had the susceptibility of CHD patients with diabetes. Our study indicated that GAS5/METTL14/ESR1 polymorphisms were associated with CHD risk, which might provide a new understanding of CHD in a Chinese population.

Summary Heterotopic ossification (HO) is a form of pathological cell-fate change of mesenchymal stem/precursor cells (MSCs) that occurs following traumatic injury, limiting range of motion in extremities and causing pain. MSCs have been shown to differentiate to form bone; however, their lineage and aberrant processes after trauma are not well understood. Utilizing a well-established mouse HO model and inducible lineage-tracing mouse (Hoxa11-CreERT2;ROSA26-LSL-TdTomato), we found that Hoxa11-lineage cells represent HO progenitors specifically in the zeugopod. Bioinformatic single-cell transcriptomic and epigenomic analyses showed Hoxa11-lineage cells are regionally restricted mesenchymal cells that, after injury, gain the potential to undergo differentiation toward chondrocytes, osteoblasts, and adipocytes. This study identifies Hoxa11-lineage cells as zeugopod-specific ectopic bone progenitors and elucidates the fate specification and multipotency that mesenchymal cells acquire after injury. Furthermore, this highlights homeobox patterning genes as useful tools to trace region-specific progenitors and enable location-specific gene deletion., Graphical Abstract, Highlights • Lineage tracing, single-cell RNA-seq and single cell ATAC enable cell specific analysis of in vivo cell fate • Hoxa11 lineage marks distinct mesenchymal precursors in the zeugopod • Hoxa11 lineage mesenchymal precursors undergo an aberrant cell fate change towards ectopic bone and cartilage, Here, Pagani et al. use the Hoxa11 patterning gene, expressed exclusively in the limb zeugopod, to demonstrate by bioinformatic single cell transcriptomic, epigenomic analyses, and ex vivo confocal microscopy that extra-skeletal cells originating from Hoxa11 lineage cells are a population of mesenchymal cells with the ability to undergo aberrant differentiation towards chondrocytes, osteoblasts or adipocytes after injury

The homologous leucine zipper/EF-hand-containing transmembranes (LETMs) are highly conserved across a broad range of eukaryotic organisms. The LETM functional characteristics involved in biological process have been identified primarily in animals, but little is known about the LETM biological function mode in plants. Based on the results of the current investigation, the GhLETM1 gene crucially affects filament elongation and anther dehiscence of the stamen in cotton. Both excessive and lower expression of the GhLETM1 gene lead to defective stamen development, resulting in shortened filaments and indehiscent anthers with pollen abortion. The results also showed that the phenotype of the shortened filaments was negatively correlated with anther defects in the seesaw model under the ectopic expression of GhLETM1. Moreover, our results notably indicated that the gene requires accurate expression and exhibits a sensitive dose effect for its proper function. This report has important fundamental and practical significance in crop science, and has crucial prospects for genetic engineering of new cytoplasmic male sterility lines and breeding of crop hybrid varieties.

Background: Cardiac dysfunction is a major component of sepsis-induced multiorgan failure in critical care units. Changes in cardiac autophagy and its role during sepsis pathogenesis have not been clearly defined. Targeted autophagy-based therapeutic approaches for sepsis are not yet developed. Methods: Beclin-1-dependent autophagy in the heart during sepsis and the potential therapeutic benefit of targeting this pathway were investigated in a mouse model of lipopolysaccharide (LPS)-induced sepsis. Results: LPS induced a dose-dependent increase in autophagy at low doses, followed by a decline that was in conjunction with mammalian target of rapamycin activation at high doses. Cardiac-specific overexpression of Beclin-1 promoted autophagy, suppressed mammalian target of rapamycin signaling, improved cardiac function, and alleviated inflammation and fibrosis after LPS challenge. Haplosufficiency for beclin 1 resulted in opposite effects. Beclin-1 also protected mitochondria, reduced the release of mitochondrial danger-associated molecular patterns, and promoted mitophagy via PTEN-induced putative kinase 1-Parkin but not adaptor proteins in response to LPS. Injection of a cell-permeable Tat-Beclin-1 peptide to activate autophagy improved cardiac function, attenuated inflammation, and rescued the phenotypes caused by beclin 1 deficiency in LPS-challenged mice. Conclusions: These results suggest that Beclin-1 protects the heart during sepsis and that the targeted induction of Beclin-1 signaling may have important therapeutic potential.

Mesenchymal stem cells (MSCs) derived from different tissues may aid in the regeneration of radiation-induced organ lesions; however, the radiation responses of human MSCs from different sources are unknown. In our study, a comparison of the results from cell proliferation, apoptosis, cell cycle, DNA damage, and DNA repair assays consistently showed that MSCs derived from adipose tissue possess a significantly stronger radiation resistance capacity than MSCs derived from umbilical cord and gingival, which is accompanied by a higher level of phosphorylated signal transducer and activator of transcription 3 (Stat3) expression. This reminds us Stat3 could be a potential biomarker of radiation resistance. These findings provide a better understanding of radiation-induced biologic responses in MSCs and may lead to the development of better strategies for stem cell treatment and cancer therapy.

Introduction: Inappropriate activation of Wnt/β-catenin signaling plays a role in some cancers. β-catenin (β-cat) levels in the cell can be regulated by a cadherin-mediated sequestration into membrane-bound and free cytosolic pools, with the later translocating to the nucleus and driving TCF-mediated transcriptional activity following Wnt signal transduction. While sequencing has shown that MM lacks the mutations that typically lead to constitutive β-cat activation seen in other cancers, we and others have demonstrated that Wnt/β-catenin signaling is nonetheless activated in MM and can regulate MM growth. The mechanism driving β-cat stabilization and activation in MM is unclear. E- and N-cadherin (N-cad) expression is elevated in MM compared to plasma cells from healthy donors. We hypothesized that that cadherins can regulate Wnt/β-catenin signaling in MM. Materials and Methods: We detected different forms of β-cat expression in a panel of human MM cell lines (HMCLs) and CD138 PC from MM patients by several approaches. Cadherin gain- or loss-of-function MM models were produced by expressing wild-type N-cad in MMS1 and ARP1 (lack endogenous N-cadherin expression) using a lentiviral system to create stable cell lines (N-Cad/MMS1 and N-cad/ARP1) and empty vector control (EV/MMS1, and EV-ARP1). We knocked down N-cadherin in the JJN3 cell line expressing high level of endogenous N-cadherin using shRNA specific for N-cad (shNcad/JJN3) or scrambled control shRNA (shCont/JJN3) by lentiviral-mediated transfection. We used a TCF reporter system to evaluate β-cat transcriptional activity as previously described. Results: We surveyed 25 HMCLs and CD138-selected plasma cells from 72 newly diagnosed MM for active β-cat with an antibody that specifically recognizes the unphosphorylated active form of β-cat. Higher levels of cytosolic and/or nuclear β-cat protein were seen in 13 of 25 (52%) HMCLs and 36 of 72 (50%) primary MM PC. Correlation of β-cat protein levels with global mRNA expression levels in primary PC revealed significant correlation with only one gene, CDH2 (N-cad). Remarkably, those primary MM with high β-cat levels but low CDH2 levels expressed high levels of E-cadherin/CHD1 mRNA. This posed the question of whether CDH2 is a direct target of TCF/β-cat transcriptional activity or whether high levels of CDH2 lead to increased levels of β-cat protein via sequestration. Both CDH2 mRNA and protein were correlated with β-cat protein but not β-cat mRNA in 23/25 HMCLs. Co-immunoprecipitation revealed that N-cad and β-cat complexes could be identified in HMCLs and primary MM. Consistent with N-cad-mediated stabilization of β-cat both total and unphosphorylated β-cat levels and TCF activity were significantly elevated in N-cad/MMS1 and N-Cad/ARP1 cells relative to controls. In contrast, shRNA mediated knockdown of N-cad led to a loss of both N-cad and β-cat protein levels and TCF-dependent transcription activity relative to controls. These findings provide evidence that β-cat/TCF signaling can be regulated by N-cad in MM. CDH2 mRNA is significantly elevated in the MS and HY subgroups of MM. To search for a potential mechanism of CDH2 transcriptional regulation in MS MM, we compared TCF activity and β-cat protein levels in MS versus non-MS HMCLs. TCF activity and active β-cat were elevated in MS versus non-MS forms of MM and B-cell lymphoma lacking N-cadherin. To determine if MMSET is required to up-regulate N-cad expression, we depleted the full-length MMSET protein in KMS11 cells. The results revealed a dramatic loss of total and unphosphorylated β-cat protein, but not mRNA, and loss of both CDH2 mRNA and protein relative to controls. These data suggest that MMSET can regulate the transcription of the CDH2 gene. MMS1 and ARP1 cells stably expressing N-cad exhibited enhanced adhesion to bone marrow stromal cells and decreased sensitivity to bortezomib (Bzb). In contrast, blocking N-cadherin-mediated adhesion by CDH2 shRNA increased sensitivity to Bzb. These results suggests that N-cad/β-cat complexes can contribute to adhesion-mediated drug resistance in MM. Conclusion: Taken together, these findings establish that β-cat is stabilized by N-cadherin-, and likely E-cadherin-, adhesins junction formation in MM. This in turn leads to an increased pool of β-cat that can drive TCF transcriptional activation as well participate in cadherin-mediated cell adhesion and drug resistance. Disclosures Davies: Amgen: Consultancy, Honoraria; BMS: Consultancy, Honoraria; Abbvie: Consultancy, Honoraria; Takeda: Consultancy, Honoraria; Janssen: Consultancy, Honoraria; Roche: Consultancy, Honoraria. Morgan: BMS: Membership on an entity's Board of Directors or advisory committees; Jansen: Membership on an entity's Board of Directors or advisory committees; Karyopharm: Membership on an entity's Board of Directors or advisory committees; Oncopeptides: Membership on an entity's Board of Directors or advisory committees. Walker: Bristol Myers Squibb: Research Funding; Sanofi: Speakers Bureau.

Efforts to identify and target glioblastoma (GBM) drivers have primarily focused on receptor tyrosine kinases (RTKs). Clinical benefits, however, have been elusive. Here, we identify a SRY-related box 2 (SOX2) transcriptional regulatory network that is independent of upstream RTKs and is capable of driving glioma initiating cells. We identified oligodendrocyte lineage transcription factor 2 (OLIG2) and zinc finger E-box binding homeobox 1 (ZEB1) as potential SOX2 targets, which are frequently co-expressed irrespective of driver mutations. In murine glioma models, we show that different combinations of tumor suppressor and oncogene mutations can activate Sox2, Olig2, and Zeb1 expression. We demonstrate that ectopic co-expression of the three transcription factors can transform tumor suppressor deficient astrocytes into glioma initiating cells in the absence of an upstream RTK oncogene. Finally, we demonstrate that the transcriptional inhibitor mithramycin downregulates SOX2 and its target genes, resulting in markedly reduced proliferation of GBM cells in vivo.

Objective: To detect the initiation of swallowing in patients with disorders of consciousness (DOC) as well as the relationship between the initiation of swallowing and the prognosis of DOC patients. Methods: Nineteen DOC patients were included in this study, and a self-controlled trial compared five different stimuli. The five different stimuli were as follows: (1) one command, as recommended by the Coma Recovery Scale-Revised (CRS-R), which was "open your mouth"; (2) placing a spoon in front of the patient's mouth without a command; (3) placing a spoon filled with water in front of the patient's mouth without a command; (4) one command-"there is a spoon; open your mouth"-with a spoon in front of the patient's mouth; (5) one command, "there is a spoon with water; open your mouth," with a spoon filled with water in front of the patient's mouth. All 19 patients were given these five stimuli randomly, and any one of the commands was presented four times to a patient, one at a time, at 15-s intervals. The sensitivity and specificity of the initiation of swallowing in detecting conscious awareness were determined. Results: None of the patients responded to the first four stimuli. However, six patients showed initiated swallowing toward the fifth stimulus. Among those six, five patients showed improvement in their consciousness state 6 months later. The sensitivity and specificity of the initiation of swallowing for DOC patients was 83.33% [95% CIs (36%, 100%)] and 92.31% [95% CIs (64%, 100%)], respectively. Conclusions: The initiation of swallowing can be an early indication of conscious behavior and can likely provide evidence of conscious awareness. Clinical Trial Registration: www.ClinicalTrials.gov, identifier: NCT03508336; Date of registration: 2018/4/16.

Background Coronary heart disease (CHD) is a common chronic inflammatory disease. Interleukin (IL)-7/IL-7R has been reported to be involved in the development of CHD. However, the relationship between IL-7/7R genetic polymorphisms and CHD among the Han Chinese population remains unclear. Methods To examine whether IL-7/7R variants contributed to CHD, six single-nucleotide polymorphisms (SNPs) were genotyped by using the Agena MassARRAY platform in 499 CHD patients and 496 controls. Logistic regression was used to calculate the odds ratios (ORs) and 95% confidence intervals (CIs). The linkage disequilibrium was analyzed using Haploview software. The association between clinical parameters and IL-7/7R polymorphisms was determined by a one-way ANOVA. Results IL-7R rs969129 G (OR = 1.20, 95% CI: 1.00–1.43, p = 0.047) allele and GG (OR = 1.45, 95% CI: 1.01–2.08, p = 0.044) genotype carriers had a higher risk for CHD. IL-7R haplotype “ACAG” (OR = 1.43, 95% CI: 1.09–1.87, p = 0.010) conferred an increased CHD risk. Rs969129, rs6451231, and rs117173992 were related to CHD susceptibility in males and/or the subgroup of individuals aged >61 years. IL-7R rs969129, rs10053847, rs6451231, and rs118137916 variants were associated with diabetes in patients with CHD. Moreover, rs969129, rs6451231, and rs117173992 were associated with high-density lipoprotein cholesterol (HDL-C) concentrations, whereas rs118137916 and rs10053847 were associated with low-density lipoprotein cholesterol (LDL-C) levels (p Conclusion IL-7/7R variants were related to the genetic predisposition of CHD in the Chinese Han population. These findings increase our knowledge regarding the effect of IL-7/7R on CHD.

Sepsis is a leading cause of death in intensive care units, and cardiac dysfunction is an identified serious component of the multi-organ failure associated with this critical condition. This review summarized the current discoveries and hypothesizes of how autophagy changes in the heart during sepsis and the underlying mechanisms. Recent investigations suggest that specific activation of autophagy initiation factor Beclin-1 has a potential to protect cardiac mitochondria, attenuate inflammation, and improve cardiac function in sepsis. Accordingly, pharmacological interventions targeting this pathway have a potential to become an effective approach to control sepsis outcomes. The role of autophagy during sepsis pathogenesis has been under intensive investigation in recent years. It is expected that developing therapeutic approaches with specificities targeting at autophagy regulatory factors may provide new opportunities to alleviate organ dysfunction caused by maladaptive autophagy during sepsis.

Berberine (BBR) is an isoquinoline alkaloid with hepatoprotective, anti-obesity, antibacterial and anti-inflammatory properties. The present study investigated berberine supplementation in high starch diet on growth performance, antioxidative status, immune parameters, and ammonia stress response of black sea bream (Acanthopagrus schlegelii). Three isonitrogenous diets were prepared as Con: control diet (14% starch), HS: high starch diet (23% starch), and HSB: high starch diet with 50 mg/kg berberine. After the 8-week feeding trial, HS and HSB groups had similar specific growth rate and weight gain, but they were significantly lower than that of the Con group (P

The aim of this study is to understand the utilization of inpatient services and its contributing factors among middle-aged and elderly females (MAEF) in less developed rural regions. Five surveys were conducted between 2006 and 2014 with rural residents of Jiangxi by stratified cluster random sampling. Participant females included only those who were aged 45 and above. Complex sampling logistics analysis was performed to analyze the effect of three factors on inpatient service. Complex sampling logistics regression analysis revealed that the probability of hospitalization for the divorced or widowed females was significantly lower than that of married ones (aOR = 0.177, p &lt, 0.05). However, the probability of early discharge was significantly higher among divorced or widowed females than married ones (aOR = 3.237, p &lt, 0.05). In addition, females with chronic diseases were more likely to be hospitalized (aOR = 3.682, p &lt, 0.05). Also, early discharge （aOR = 7.689，p &lt, 0.05) occurred among the participants who should be hospitalized but were not hospitalized occurred (aOR = 3.258，p &lt, 0.05). The continuous improvement of the new rural cooperative medical policy has promoted the utilization of inpatient services for the MAEF. Findings from this study emphasize the need to strengthen the prevention and treatment of chronic diseases among middle-aged and elderly women.

Background Behavioral assessment has been acted as the gold standard for the diagnosis of disorders of consciousness (DOC) patients. The item “Functional Object Use” in the motor function sub-scale in the Coma Recovery Scale-Revised (CRS-R) is a key item in differentiating between minimally conscious state (MCS) and emergence from MCS (EMCS). However, previous studies suggested that certain specific stimuli, especially something self-relevant can affect DOC patients’ scores of behavioral assessment scale. So, we attempted to find out if personalized objects can improve the diagnosis of EMCS in the assessment of Functional Object Use by comparing the use of patients’ favorite objects and other common objects in MCS patients. Methods Twenty-one post-comatose patients diagnosed as MCS were prospectively included. The item “Functional Object Use” was assessed by using personalized objects (e.g., cigarette, paper) and non-personalized objects, which were presented in a random order. The rest assessments were performed following the standard protocol of the CRS-R. The differences between functional uses of the two types of objects were analyzed by the McNemar test. Results The incidence of Functional Object Use was significantly higher using personalized objects than non-personalized objects in the CRS-R. Five out of the 21 MCS studied patients, who were assessed with non-personalized objects, were re-diagnosed as EMCS with personalized objects (χ2 = 5, df = 1, p

CONSORT 2010 checklist of information to include when reporting a randomised trial. (DOC 217 kb)

The details of Function Object Use with personalized object. (DOCX 25 kb)

Tissue and cell damage caused by ionizing radiation is often highly genotoxic. The swift repair of DNA damage is crucial for the maintenance of genomic stability and normal cell fitness. Long noncoding RNAs (lncRNAs) have been reported to play an important role in many physiological and pathological processes in cells. However, the exact function of lncRNAs in radiation-induced DNA damage has yet to be elucidated. Therefore, this study aimed to analyze the potential role of lncRNAs in radiation-induced DNA damage. We examined the expression profiles of lncRNAs and mRNAs in 293T cells with or without 8 Gy irradiation using high-throughput RNA sequencing. We then performed comprehensive transcriptomic and bioinformatic analyses of these sequencing results. A total of 18,990 lncRNAs and 16,080 mRNAs were detected in all samples. At 24 h post irradiation, 49 lncRNAs and 323 mRNAs were differentially expressed between the irradiation group and the control group. qRT-PCR was used to verify the altered expression of six lncRNAs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that the predicted genes were mainly involved in the histone mRNA metabolic process and Wnt signaling pathways. This study may provide novel insights for the study of lncRNAs in radiation-induced DNA damage.

Ultrafast infrared (IR) Raman spectroscopy was used to study vibrational energy in ϕ-S alkylbenzenes, where ϕ = C6H5 and substituents S were CH3- (toluene), (CH3)2CH- (isopropylbenzene, IPB), or (CH3)3C- (t-butylbenzene, TBB). Using methods described previously,1 the normal modes were classified as phenyl (ϕ), substituent (S), or global (G). IR pulses were tuned to find conditions that maximized the localization of initial CH-stretch excitations on ϕ or S. Anti-Stokes Raman spectroscopy measured transient energy content of Raman-active S, ϕ, and G modes, to determine the rates of phenyl to substituent (Φ → S) or substituent to phenyl (S → Φ) transfer during the first few picoseconds, when energy transfer was mainly intramolecular. Since phenyl CH-stretches were 90-130 cm(-1) uphill in energy from substituent CH-stretches, of interest were S → Φ processes where molecular structure and local couplings were more important than energy differences. The Φ → S process efficiencies were small and about equal with all three substituents. The S → Φ transfer efficiencies could be increased by increasing substituent size. This was opposite to what would be predicted on the basis of the larger density of states of larger substituents, and it provides a path toward controlling forward-to-backward vibrational energy transfer ratios. The S → Φ transfer efficiency is understood as resulting from an increase in the local anharmonic couplings. A heavier substituent, when vibrating, transfers energy more effectively to the phenyl group.

We introduce a novel type of three-dimensional (3D) spectroscopy to study vibrational energy transfer, where an IR pulse tunable through the CH-stretching and CD-stretching regions was used to create parent vibrational excitations in liquids and a visible probe pulse was used to generate both Stokes and anti-Stokes Raman spectra as a function of delay time. The Raman spectra determine how much vibrational excitation was present in each probed state. The three dimensions are the wavenumber of the pumped state, the wavenumber of the probed state, and the time interval. The technique was used to study nitromethane (NM) and acetonitrile (ACN) and their deuterated analogues at ambient temperature. The 3D spectra were quite complicated. Three types of artifacts due to nonlinear light scattering were observed. Along the diagonal were two fundamental CH-stretch (or CD-stretch) transitions and several weaker combination bands or overtone transitions. Because Raman spectroscopy allows us to simultaneously probe a wide wavenumber region, for every diagonal peak, there were ∼10 off-diagonal peaks. The cross-peaks at shorter delay times reveal the nature of the initial excitation by showing which lower-wavenumber excitations were produced along with the pumped CH-stretch or CD-stretch. The longer-time spectra characterized vibrational energy relaxation processes, and showed how daughter vibrations were generated by different parent excitations.

Experiments were performed on nitrobenzene liquid at ambient temperature to probe vibrational energy flow from the nitro group to the phenyl group and vice versa. The IR pump, Raman probe method was used. Quantum chemical calculations were used to sort the normal modes of nitrobenzene into three categories: phenyl modes, nitro modes, and global modes. IR wavelengths in the 2500-3500 cm(-1) range were found that best produced excitations initially localized on nitro or phenyl. Pulses at 2880 cm(-1) excited a nitro stretch combination band. Pulses at 3080 cm(-1) excited a phenyl C-H stretch plus some nitro stretch. With nitro excitation there was no detectable energy transfer to phenyl. With phenyl excitation there was no direct transfer to nitro, but there was some transfer to global modes such as phenyl-nitro stretching, so some of the vibrational amplitude on phenyl moved onto nitro. Thus energy transfer from nitro to phenyl was absent, but there was weak energy transfer from phenyl to nitro. The experimental methods described here can be used to study vibrational energy flow from one part of a molecule to another, which could assist in the design of molecules for molecular electronics and phononics. The vibrational isolation of the nitro group when attached to a phenyl moiety suggests that strongly nonthermal reaction pathways may play an important role in impact initiation of energetic materials having peripheral nitro groups.

The ability to diffusely infiltrate as single cells through the highly confined extracellular spaces of the brain parenchyma and along blood vessels is a pathological hallmark of Glioblastoma Multiforme (GBM), which makes these tumors surgically incurable. The fact that GBM cells are capable of moving through diverse brain microenvironments raises the possibility that GBM cells may possess multiple mechanisms for invasion. At present, little is known about how GBM cells gain traction and generate sufficient contractile forces to overcome the mechanical challenge of migrating through the confined interstitial spaces of the brain. The purpose of this study is to gain cellular, biochemical and biophysical insights into GBM migration. To accomplish this we generated 1-D micro-patterned arrays that approximate the topography of brain microvasculature and microfluidic devices that approximate brain interstitial spaces. High-resolution 3D time lapse imaging is then used to track actin, myosin-II and alpha-tubulin dynamics during cell migration. We found that the geometry of micro-patterned laminin lanes in the absence of chemotactic, hapatotactic or durotactic gradients, is sufficient to trigger sustained (integrin) adhesion-dependent migration that is driven by actin polymerization at the leading edge and supported by acto-myosin-II cable-like structures that extend along the ventral and lateral edges of spindle shaped cells. A similar pattern of adhesion-dependent (mesenchymal) migration was seen in 3D channels with aspect ratio (15x15um) larger than the cell. In confined (5x5um) channels, GBM cells adopted interchangeable modes of migration, waxing between blebbing and mesenchyal migration (approximately 50:50). Blebbing migration was characterized by acto-myosin-II cable-like structures that extended the length of the cell along both ventral and dorsal surfaces. In contrast, mesenchymal phase migration was characterized by a cytoskeleton organization pattern that resembled 1-D migration. Exposure of cells in confined channels to actin inhibitors (latrunculin B and CK-666) caused the cells to adopt a blebbing predominant mode of migration, while myosin-II inhibitor (blebbistatin) induced a mesenchymal predominant pattern. While neither class of inhibitors blocked migration as a single agent, the combination produced cell arrest. GBM cells were also able to migrate through confined channels in the absence of any ECM coating by adopting a blebbing exclusive pattern of migration. This observation raises the possibility that in the absence of integrin signaling, GBM cells can use hydrostatic pressure to gain traction in confined spaces. Taken together these results suggest that GBM can interchangeably adopt adhesion dependent and independent modes of migration as they encounter different sized ECM topography and variable 3-D confinement. Citation Format: James O. Nyagilo, Sara Picirrillo, Yuxiao Sun, Loan T. Bui, Nanda Regmi, Jamie L. Wright, Donald Thevalingam, Armin Soltan Zadi, Young-tae Kim, Charles Choung, Digant P. Dave, Robert Bachoo. Extracellular matrix geometry and 3D spatial confinement trigger diverse mechanisms of primary human glioblastoma cell migration. [abstract]. In: Proceedings of the AACR Special Conference on Engineering and Physical Sciences in Oncology; 2016 Jun 25-28; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2017;77(2 Suppl):Abstract nr A54.

We have previously shown that mitochondria-targeted vitamin E (Mito-Vit-E), a mtROS specific antioxidant, improves cardiac performance and attenuates inflammation in a pneumonia-related sepsis model. In this study, we applied the same approaches to decipher the signaling pathway(s) of mtROS-dependent cardiac inflammation after sepsis. Sepsis was induced in Sprague Dawley rats by intratracheal injection of S. pneumoniae. Mito-Vit-E, vitamin E or vehicle was administered 30 minutes later. In myocardium 24 hours post-inoculation, Mito-Vit-E, but not vitamin E, significantly protected mtDNA integrity and decreased mtDNA damage. Mito-Vit-E alleviated sepsis-induced reduction in mitochondria-localized DNA repair enzymes including DNA polymerase γ, AP endonuclease, 8-oxoguanine glycosylase, and uracil-DNA glycosylase. Mito-Vit-E dramatically improved metabolism and membrane integrity in mitochondria, suppressed leakage of mtDNA into the cytoplasm, inhibited up-regulation of Toll-like receptor 9 (TLR9) pathway factors MYD88 and RAGE, and limited RAGE interaction with its ligand TFAM in septic hearts. Mito-Vit-E also deactivated NF-κB and caspase 1, reduced expression of the essential inflammasome component ASC, and decreased inflammatory cytokine IL–1β. In vitro, both Mito-Vit-E and TLR9 inhibitor OND-I suppressed LPS-induced up-regulation in MYD88, RAGE, ASC, active caspase 1, and IL–1β in cardiomyocytes. Since free mtDNA escaped from damaged mitochondria function as a type of DAMPs to stimulate inflammation through TLR9, these data together suggest that sepsis-induced cardiac inflammation is mediated, at least partially, through mtDNA-TLR9-RAGE. At last, Mito-Vit-E reduced the circulation of myocardial injury marker troponin-I, diminished apoptosis and amended morphology in septic hearts, suggesting that mitochondria-targeted antioxidants are a potential cardioprotective approach for sepsis.

During mitosis, equal transmission of the duplicated chromosomes demands a strict regulation of separase, which cleaves cohesin and triggers sister chromatid separation in anaphase. Vertebrate separase is inhibited by securin and the inhibitory phosphorylation of separase. However, knockout experiments indicate that securin is dispensable and the inhibitory phosphorylation was observed only in M phase cells. This begs the question how cohesin cleavage by separase is prevented in the absence these two mechanisms. Here we show that separase is excluded from cohesin by the nuclear envelope, which forms in telophase and disassembles in mitosis. The exclusion is achieved passively by its large physical mass and may be backed up by the CRM1-dependent nuclear export. A functional NES motif is identified in separase. We demonstrated that the nuclear envelope is sufficient to prevent active separase from cleaving nuclear cohesin. We propose that the nuclear exclusion is important to prevent cohesin cleavage during interphase in the absence of securin and the phosphorylation inhibition.

Sister chromatids in mammalian cells remain attached mostly at their centromeres at metaphase because of the loss of cohesion along chromosome arms in prophase. Here, we report that Bub1 retains centromeric cohesion in mitosis of human cells. Depletion of Bub1 or Shugoshin (Sgo1) in HeLa cells by RNA interference causes massive missegregation of sister chromatids that originates at centromeres. Surprisingly, loss of chromatid cohesion in Bub1 and Sgo1 RNA-interference cells does not appear to require the full activation of separase but, instead, triggers a mitotic arrest that depends on Mad2 and Aurora B. Bub1 maintains the steady-state levels and centromeric localization of Sgo1. Therefore, Bub1 protects centromeric cohesion through Shugoshin in mitosis.

A more accurate, integrated chronostratigraphic framework is applied to the analysis of the development of Paleogene depressions in the Pearl River Mouth basin. The results of our study show that the development of these depressions was characterized by at least three rifting or basin-forming phases occurring during these periods: late Paleocene (Late Cretaceous?)-middle Eocene, late Eocene-early Oligocene, and middle Oligocene-late Oligocene. The transition from rifting stage to postrifting stage in the basin is about 10 m.y. later than the initial spreading of the South China Sea. The prologue of the spreading of the South China Sea began as early as the end of the middle Eocene. Lacustrine source rocks deposited during the basins first rifting phase are thick and of go d quality; source rocks deposited during the last two phases, which had a sharp increase in sedimentation rate, are of lesser quality, with the exception being those areas where deposits were out of reach of sediment from the northern mainland.

SummarySister chromatid separation is triggered by the separase-catalyzed cleavage of cohesin. This process is temporally controlled by cell-cycle-dependent factors, but its biochemical mechanism and spatial regulation remain poorly understood. We report that cohesin cleavage by human separase requires DNA in a sequence-nonspecific manner. Separase binds to DNA in vitro, but its proteolytic activity, measured by its autocleavage, is not stimulated by DNA. Instead, biochemical characterizations suggest that DNA mediates cohesin cleavage by bridging the interaction between separase and cohesin. In human cells, a fraction of separase localizes to the mitotic chromosome. The importance of the chromosomal DNA in cohesin cleavage is further demonstrated by the observation that the cleavage of the chromosome-associated cohesins is sensitive to nuclease treatment. Our observations explain why chromosome-associated cohesins are specifically cleaved by separase and the soluble cohesins are left intact in anaphase.