Identification of human fetal liver miRNAs by a novel method

AbstractMicroRNAs (miRNAs) are short 20–25 nucleotides RNA molecules that have been shown to regulate gene expressions in a variety of eukaryotic systems. miRNAs are widespread in eukaryotes and several hundred of miRNAs have been identified, but still a lot of miRNAs have not been detected in various eukaryotic organisms. However, it is not an easy work to clone miRNAs by traditional methods. Here, we describe the identification of 27 miRNAs from a human fetal liver cDNA library by a novel cloning method. Low molecular weight RNA fraction (⩽200nt) from fetal liver tissue was extracted, and polyadenylated by poly(A) polymerase. A 5′ RNA adaptor was ligated to poly(A)-tailed RNA using T4 RNA ligase. After reverse transcription, the cDNA was amplified by PCR with two adaptor primers. The PCR product with a size about 109bp was recovered and cloned into T vector. After sequencing, database searching, and expression profiling, 5 novel miRNAs were discovered among other 22 known miRNAs in human fetal liver. These finding indicate that a large diverse population of miRNAs may function to regulate gene expression in hepatocyte

Overexpression of Toll-Like Receptor 4 Contributes to Phagocytosis of Salmonella Enterica Serovar Typhimurium via Phosphoinositide 3-Kinase Signaling in Sheep

Background/Aims: Phagocytosis of bacteria by monocytes/macrophages can trigger the immune response and the clearance of bacteria. This innate immune response involves Toll-like receptor 4 (TLR4). However, much remains unknown about the mechanism of TLR4-regulated phagocytosis of Salmonella enterica serovar Typhimurium (S. typhimurium) within sheep monocytes/macrophages. Here, we aimed to address these knowledge gaps by infecting transgenic sheep overexpressing TLR4 with S. typhimurium and examining the phagocytic mechanisms involved. Methods: Transgenic sheep were generated by microinjection of the constructed plasmids into fertilized eggs. Monocytes/macrophages isolated from sheep blood were stimulated with LPS and S. typhimurium. Phagocytosis-related factor expression, phagocytic ability, and adhesion were then determined. TLR4/phosphatidylinositide 3-kinase (PI3K) signaling was inhibited to investigate if this pathway is involved in changes in bacterial internalization in sheep. Results: We found that TLR4 overexpression effectively activated the PI3K signaling pathway and upregulated the expression of scavenger receptors. Additionally, actin polymerization and adhesive capacity were both enhanced in TLR4-overexpressing sheep monocytes/macrophages. TLR4 inhibition decreased S. typhimurium phagocytosis by reducing the actin polymerization and adhesive capacity of cells. Furthermore, inhibition of PI3K markedly impaired TLR4-dependent phagocytosis by restraining actin polymerization and scavenger receptor expression and reduced the adhesive capacity of the monocytes/macrophages. Conclusion: Our findings indicate that overexpression of TLR4 enhances phagocytosis through PI3K signaling and the subsequent activation of actin polymerization and scavenger receptors in sheep monocytes/macrophages infected with S. typhimurium

Degraded Synergistic Recruitment of sEMG Oscillations for Cerebral Palsy Infants Crawling

Background: Synergistic recruitment of muscular activities is a generally accepted mechanism for motor function control, and motor dysfunction, such as cerebral palsy (CP), destroyed the synergistic electromyography activities of muscle group for limb movement. However, very little is known how motor dysfunction of CP affects the organization of the myoelectric frequency components due to the abnormal motor unit recruiting patterns.Objectives: Exploring whether the myoelectric activity can be represented with synergistic recruitment of surface electromyography (sEMG) frequency components; evaluating the effect of CP motor dysfunction on the synergistic recruitment of sEMG oscillations.Methods: Twelve CP infants and 17 typically developed (TD) infants are recruited for self-paced crawling on hands and knees. sEMG signals have been recorded from bilateral biceps brachii (BB) and triceps brachii (TB) muscles. Multi-scale oscillations are extracted via multivariate empirical mode decomposition (MEMD), and non-negative matrix factorization (NMF) method is employed to obtain synergistic pattern of these sEMG oscillations. The coefficient curve of sEMG oscillation synergies are adopted to quantify the time-varying recruitment of BB and TB myoelectric activity during infants crawling.Results: Three patterns of sEMG oscillation synergies with specific frequency ranges are extracted in BB and TB of CP or TD infants. The contribution of low-frequency oscillation synergy of BB in CP group is significantly less than that in TD group (p < 0.05) during forward swing phase for slow contraction; however, this low-frequency oscillation synergy keep higher level during the backward swing phase crawling. For the myoelectric activities of TB, there is not enough high-frequency oscillation recruitment of sEMG for the fast contraction in propulsive phase of CP infants crawling.Conclusion: Our results reveal that, the myoelectric activities of a muscle can be manifested as sEMG oscillation synergies, and motor dysfunction of CP degrade the synergistic recruitment of sEMG oscillations due to the impaired CNS regulation and destroyed MU/muscle fiber. Our preliminary work suggests that time-varying coefficient curve of sEMG oscillation synergies is a potential index to evaluate the abnormal recruitment of electromyography activities affected by CP disorders

Glycyrrhizin Treatment Facilitates Extinction of Conditioned Fear Responses After a Single Prolonged Stress Exposure in Rats

Background/Aims: Impaired fear memory extinction is widely considered a key mechanism of post-traumatic stress disorder (PTSD). Recent studies have suggested that neuroinflammation after a single prolonged stress (SPS) exposure may play a critical role in the impaired fear memory extinction. Studies have shown that high mobility group box chromosomal protein 1 (HMGB-1) is critically involved in neuroinflammation. However, the role of HMGB-1 underlying the development of impairment of fear memory extinction is still not known. Methods: Thus, we examined the levels of HMGB-1 in the basolateral amygdala (BLA) following SPS using Western blot and evaluated the levels of microglia and astrocytes activation in the BLA after SPS using immunohistochemical staining. We then examined the effects of pre-SPS intra-BLA administration of glycyrrhizin, an HMGB1 inhibitor, or LPS-RS, a competitive TLR4 antagonist, on subsequent post-SPS fear extinction. Results: We found that SPS treatment prolonged the extinction of contextual fear memory after the SPS. The impairment of SPS-induced extinction of contextual fear memory was associated with increased HMGB1 and Toll-like receptor 4 (TLR4) levels in the BLA. Additionally, the impairment of SPS-induced extinction of contextual fear memory was associated with increased activation of microglia and astrocyte in the BLA. Intra-BLA administrations of glycyrrhizin (HMGB-1 inhibitor) or LPS-RS (TLR4 antagonist) can prevent the development of SPS-induced fear extinction impairment. Conclusion: Taken together, these results suggested that SPS treatment may not only produce short term effects on the HMGB1/TLR4-mediated pro-inflammation, but alter the response of microglia and astrocytes to the exposure to fear associated contextual stimuli

Analysis on the Noise for the Different Gearboxes of the Heavy Truck

In order to locate the excessive noise caused by gearbox noise when a heavy vehicle is accelerating, the noise and vibration on the key parts of the gearbox were tested and analyzed, and the peak noise frequency of radiating from the subbox of gearbox was found, which is the same as the peak frequency of pass-by noise. Aiming at the larger gear noise of the gearbox subbox, the experiments of pass-by noise by the two ways of the different speed-ratio gearbox, and the transformation of vice-box with helical gear, the pass-by noise was reduced by 5.6 and 3.9 dB(A), and it was made to reach the less GB limit-value. Meanwhile, the effect of the different-ratio gearbox on the vehicle noise was analyzed. So, the methods provide a practical basis for the vehicle noise and the fault diagnosis of gearbox

A Comprehensive Immunologic Portrait of Triple-Negative Breast Cancer

Triple-negative breast cancer (TNBC) is a high-risk malignancy due to its high capacity for invasion and lack of targeted therapy. Immunotherapy continues to demonstrate efficacy in a variety of cancers, and thus may be a promising strategy for TNBC given the limited therapeutic options currently available for TNBC. In this study, we performed an exhaustive analysis of immunogenic signatures in TNBC based on 2 large-scale breast cancer (BC) genomic data. We compared enrichment levels of 26 immune cell activities and pathways among TNBC, non-TNBC, and normal tissue, and within TNBCs of different genotypic or phenotypic features. We found that almost all analyzed immune activities and pathways had significantly higher enrichment levels in TNBC than non-TNBC. Elevated enrichment of these immune activities and pathways was likely to be associated with better survival prognosis in TNBC. This study demonstrated that TNBC likely exhibits the strongest immunogenicity among BC subtypes, and thus warrants the immunotherapeutic option for TNBC

Immunogenomics Analysis Reveals that TP53 Mutations Inhibit Tumor Immunity in Gastric Cancer

Although immunotherapy continues to demonstrate efficacy in a variety of refractory cancers, currently, no any immunotherapeutic strategy is clinically used for gastric cancer (GC) except its microsatellite instable subtype. Thus, it is important to identify molecular biomarkers for predicting the responders to GC immunotherapy. TP53 mutations frequently occur in GC and are associated with unfavorable clinical outcomes in GC. We performed a comprehensive characterization of the associations between TP53 mutations and immune activities in GC based on two large-scale GC cancer genomics data. We compared expression and enrichment levels of 787 immune-related genes and 23 immune gene-sets among TP53-mutated GCs, TP53‐wildtype GCs, and normal tissue, and explored the correlations between p53-mediated pathways and immune activities in GC. Strikingly, almost all analyzed immune gene-sets were significantly downregulated in enrichment levels in TP53-mutated GCs compared to TP53‐wildtype GCs. These less active immune pathways and cell types in TP53-mutated GCs included 15 immune cell types and function, tumor-infiltrating lymphocytes, regulatory T cells, immune checkpoint, cytokine and cytokine receptor, human leukocyte antigen, pro‐inflammatory, and parainflammation. Moreover, we identified a number of p53-mediated pathways and proteins that were significantly associated with immune activities in GC. Furthermore, we demonstrated that the TP53 mutation itself could result in the depressed immune activities in GC and other cancer types. We revealed that chromosomal instability was an important mechanism for the depressed tumor immunity in TP53-mutated cancers. Finally, we showed that immune cell infiltration and immune activities were likely positively associated with survival prognosis in GC. Our findings suggest that p53 may play an important role in activating tumor immunity in GC and other cancer types and that the TP53 mutation status could be useful in stratifying cancer patients responsive to a certain immunotherapy

Research on the Establishment of Provincial Characteristic Scenic Lines Based on GIS

China is entering a stage of rapid development. To ensure strategic development, more regions have begun to integrate and reconstruct regional spaces by strengthening their regional cooperation and focusing on top-level design. The scenic line is a physical space that integrates ecological landscape resources, cultural carriers, and industrial foundations into regional spaces. Its construction is of considerable importance for aggregating functions and supporting regional integration. Sichuan, China, has some of the most abundant bamboo resources worldwide, and the bamboo scenic line has Chinese characteristics. This study takes 12 areas suitable for bamboo growth in Sichuan Province as research objects using GIS technology combined with methods such as suitability evaluation. An ecological base layer, landscape pattern layer, facility foundation layer, and industrial development layer were developed as the four element layers, along with their influencing factors. The weights of the factors were assigned using the entropy method, and the cost path was analyzed for the resistance side, branch point, and context. A suitability evaluation system was constructed for the scenic line, and a provincial organic development pattern of “one point, two axes and three belts” was formed for the bamboo scenic line, which can provide guidance for planning and design

Increased glycolysis correlates with elevated immune activity in tumor immune microenvironmentResearch in context section

Background: Prior studies showed that tumor glycolysis and tumor immune evasion are interdependent. However, a systematic investigation of the association between tumor glycolysis and tumor immunity in various cancers remains lacking. Methods: Using the Cancer Genome Atlas (TCGA) datasets, we explored the association between glycolytic activity and immune signatures in 14 cancer types. We also explored the associations between glycolytic activity and tumor immunity associated genetic features, including PD-L1 expression, tumor mutation burden (TMB), and tumor aneuploidy. Moreover, we performed in vitro experiments to verify some findings from bioinformatics analysis. Furthermore, we explored the association between tumor glycolytic activity and immunotherapy response. Findings: Glycolytic activity was likely correlated with active immune signatures in various cancers and highly glycolytic tumors presented an immune-stimulatory tumor microenvironment. Compared to TMB and aneuploidy, glycolytic activity was a stronger and more consistent predictor for immune signatures in diverse cancers. Both computational and experimental analyses showed that glycolysis could increase PD-L1 expression in tumor. Glycolytic activity had a strong correlation with apoptosis which was a strong positive predictor for immune signatures, suggesting that apoptosis could be an important medium connecting glycolytic activity with immune activity in cancer. Finally, highly glycolytic tumors exhibited a better immunotherapy response and a favorable survival in the immunotherapy setting. Interpretation: Tumor glycolysis may increase tumor immunity in diverse cancers. Glycolytic activity enhances PD-L1 expression on tumor cells and thus promotes anti-PD-1/PD-L1 immunotherapy response. Thus, the tumor glycolytic activity could be a predictive biomarker for immunotherapy response in diverse cancers. Fund: This work was supported by the China Pharmaceutical University (grant numbers 3150120001, 2632018YX01 to XW). Keywords: Tumor glycolysis, Tumor immunity, Tumor immune microenvironment, PD-L1 expression, Tumor immunotherap