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ObjectiveTo investigate the impact of different component ratios and mechanical stiffness of the gelatin-sodium alginate composite hydrogel scaffold, fabricated through 3D bioprinting, on the viability and functionality of chondrocytes.MethodsThree different concentrations of hydrogel, designated as low, medium, and high, were prepared. The rheological properties of the hydrogel were characterized to optimize printing parameters. Subsequently, the printability and shape fidelity of the cell-loaded hydrogel scaffolds were statistically evaluated, and the chondrocyte viability was observed. Dynamic mechanical analysis was conducted to measure the modulus, thereby assessing the scaffold’s stiffness. Following a 21-day culture period, RT-PCR, histological staining, and immunostaining were employed to assess chondrocyte activity, chondrosphere aggregates formation, and cartilage matrix production.ResultsBased on rheological analysis, optimal printing temperatures for each group were determined as 27.8°C, 28.5°C, and 30°C. The optimized printing parameters could ensure the molding effect of the scaffolds on the day of printing, with the actual grid area of the scaffolds was close to the theoretical grid area. And the scaffolds exhibited good cell viability (93.24% ± 0.99%, 92.04% ± 1.49%, and 88.46% ± 1.53%). After 7 days of culture, the medium and high concentration groups showed no significant change in grid area compared to the day of printing (p > 0.05), indicating good morphological fidelity. As the hydrogel’s bicomponent ratio increased, both the storage modulus and loss modulus increased, while the loss factor remained relatively constant. The highest number of chondrocytes-formed chondrosphere aggregates in the medium concentration group was observed by light microscopy. RT-PCR results indicated significantly higher expression levels of chondrogenic genes SOX9, Agg, and Col-II in the low and medium concentration groups compared to the high concentration group (p < 0.05). Histological staining results showed that the middle concentration group formed the highest number of typical cartilage lacunae.ConclusionThe aforementioned results indicate that in 3D bioprinted cell-loaded GA-SA composite hydrogel scaffolds, the scaffolds with the composition ratio (10:3) and mechanical stiffness (∼155 kPa) exhibit sustained morphological fidelity, effectively preserve the hyaline phenotype of chondrocytes, and are more conducive to cartilage regeneration.

Vascular stents have many applications in treating arterial stenosis and other vascular-related diseases. The ideal vascular stent for clinical application should have radial support and axial bending mechanical properties that meet the requirements of vascular deformation coordination. The materials used for vascular stents implanted in the human body should have corresponding biocompatibility to ensure that the stents do not cause coagulation, hemolysis, and other reactions in the blood. This study fabricated four types of vascular stents, including inner hexagon, arrowhead, quadrilateral, and outer hexagonal, using fused filament fabrication technology and thermoplastic polyurethane (TPU) as materials. By evaluating the effects of edge width and wall thickness on the radial support and axial bending performance, it was found that the inner hexagonal stent exhibited the best radial support and axial bending performance under the same conditions. The design and fabrication of vascular stents based on 3D printing technology have promising application prospects in personalized customized vascular repair therapy.

Articular osteochondral (OC) defects are a global clinical problem characterized by loss of full-thickness articular cartilage with underlying calcified cartilage through to the subchondral bone. While current surgical treatments can relieve pain, none of them can completely repair all components of the OC unit and restore its original function. With the rapid development of three-dimensional (3D) printing technology, admirable progress has been made in bone and cartilage reconstruction, providing new strategies for restoring joint function. 3D printing has the advantages of fast speed, high precision, and personalized customization to meet the requirements of irregular geometry, differentiated composition, and multi-layered boundary layer structures of joint OC scaffolds. This review captures the original published researches on the application of 3D printing technology to the repair of entire OC units and provides a comprehensive summary of the recent advances in 3D printed OC scaffolds. We first introduce the gradient structure and biological properties of articular OC tissue. The considerations for the development of 3D printed OC scaffolds are emphatically summarized, including material types, fabrication techniques, structural design and seed cells. Especially from the perspective of material composition and structural design, the classification, characteristics and latest research progress of discrete gradient scaffolds (biphasic, triphasic and multiphasic scaffolds) and continuous gradient scaffolds (gradient material and/or structure, and gradient interface) are summarized. Finally, we also describe the important progress and application prospect of 3D printing technology in OC interface regeneration. 3D printing technology for OC reconstruction should simulate the gradient structure of subchondral bone and cartilage. Therefore, we must not only strengthen the basic research on OC structure, but also continue to explore the role of 3D printing technology in OC tissue engineering. This will enable better structural and functional bionics of OC scaffolds, ultimately improving the repair of OC defects.

Abstract Background Aberrant RNA editing of adenosine-to-inosine (A-to-I) has been linked to multiple human cancers, but its role in intrahepatic cholangiocarcinoma (iCCA) remains unknown. We conducted an exome-wide investigation to search for dysregulated RNA editing that drive iCCA pathogenesis. Methods An integrative whole-exome and transcriptome sequencing analysis was performed to elucidate the RNA editing landscape in iCCAs. Putative RNA editing sites were validated by Sanger sequencing. In vitro and in vivo experiments were used to assess the effects of an exemplary target gene Kip1 ubiquitination-promoting complex 1 (KPC1) and its editing on iCCA cells growth and metastasis. Crosstalk between KPC1 RNA editing and NF-κB signaling was analyzed by molecular methods. Results Through integrative omics analyses, we revealed an adenosine deaminases acting on RNA 1A (ADAR1)-mediated over-editing pattern in iCCAs. ADAR1 is frequently amplified and overexpressed in iCCAs and plays oncogenic roles. Notably, we identified a novel ADAR1-mediated A-to-I editing of KPC1 transcript, which results in substitution of methionine with valine at residue 8 (p.M8V). KPC1 p.M8V editing confers loss-of-function phenotypes through blunting the tumor-suppressive role of wild-type KPC1. Mechanistically, KPC1 p.M8V weakens the affinity of KPC1 to its substrate NF-κB1 p105, thereby reducing the ubiquitinating and proteasomal processing of p105 to p50, which in turn enhances the activity of oncogenic NF-κB signaling. Conclusions Our findings established that amplification-driven ADAR1 overexpression results in overediting of KPC1 p.M8V in iCCAs, leading to progression via activation of the NF-κB signaling pathway, and suggested ADAR1-KPC1-NF-κB axis as a potential therapeutic target for iCCA.

The Late Pleistocene-Holocene climate fluctuations have greatly influenced the phylogeographic structure and historical dynamics of many marine organisms in the western Pacific marginal seas. Here, we investigated the impact of Pleistocene glacial-interglacial cycles on the phylogeographic structure and demographic dynamics of Decapterus maruadsi, an economically important fish along the coast of the East China Sea (ECS) and northern South China Sea (NSCS). We obtained 430 concatenated sequences (Cyt b + control region, 1548–1554 bp) of D. maruadsi, including 246 newly sampled from the ECS and 184 previously determined from the NSCS. Genetic structure and phylogenetic analysis demonstrated a lack of significant population structure among 16 populations. Moreover, there was no significant differentiation among populations from Chinese coastal waters and northern Vietnam. Neutrality tests, unimodal mismatch distributions, Bayesian skyline plots, and the star-like haplotype networks all indicated a recent demographic expansion for D. maruadsi population during the Late Pleistocene-Holocene, explaining the low genetic diversity in D. maruadsi along the southeast coast of China. Notably, phylogenetic analyses and net genetic distances based on Cyt b jointly confirmed that 57 Cyt b haplotypes identified as D. maruadsi from the previously defined Sundaland-Rosario-Ranong clade actually represented D. russelli. These results not only reveal the complex effects of Pleistocene-Holocene climate fluctuations on the phylogeographic structure and demographic history of D. maruadsi but also provide useful genetic information for the management of genetic resources.

Background: Ischemic stroke is a highly complex disorder. This study aims to identify novel methylation changes in ischemic stroke.Methods: We carried out an epigenome-wide study of ischemic stroke using an Infinium HumanMethylation 850K array (cases:controls = 4:4). 10 CpG sites in 8 candidate genes from gene ontology analytics top-ranked pathway were selected to validate 850K BeadChip results (cases:controls = 20:20). We further qualified the methylation level of promoter regions in 8 candidate genes (cases:controls = 188:188). Besides, we performed subgroup analysis, dose-response relationship and diagnostic prediction polygenic model of candidate genes.Results: In the discovery stage, we found 462 functional DNA methylation positions to be associated with ischemic stroke. Gene ontology analysis highlighted the “calcium-dependent cell-cell adhesion via plasma membrane cell adhesion molecules” item, including 8 candidate genes (CDH2/PCDHB10/PCDHB11/PCDHB14/PCDHB16/PCDHB3/PCDHB6/PCDHB9). In the replication stage, we identified 5 differentially methylated loci in 20 paired samples and 7 differentially methylated genes (CDH2/PCDHB10/PCDHB11/PCDHB14/PCDHB16/PCDHB3/PCDHB9) in 188 paired samples. Subgroup analysis showed that the methylation level of above 7 genes remained significantly different in the male subgroup, large-artery atherosclerosis subgroup and right hemisphere subgroup. The methylation level of each gene was grouped into quartiles, and Q4 groups of the 7 genes were associated with higher risk of ischemic stroke than Q1 groups (p < 0.05). Besides, the polygenic model showed high diagnostic specificity (0.8723), sensitivity (0.883), and accuracy (0.8777).Conclusion: Our results demonstrate that DNA methylation plays a crucial part in ischemic stroke. The methylation of these 7 genes may be potential diagnostic biomarker for ischemic stroke.

Computer vision is an important part of the autonomous vehicles to gain the perception of the surrounding environment. In order to satisfy the adaptability of lane detection system under different illumination and different road conditions, an effective lane detection method based on image classification and hybrid isomeric operators is proposed. Camera correction matrix and distortion coefficient are obtained by using checkerboard grid images, and they will be cached to be shared with subsequent image streams to improve the speed of real-time lane detection. In the process of preprocessing, the images under different illumination conditions are classified and processed, and the inverse perspective transformation is used to deal with the 2-D image flow so that the image flow has a certain depth of data. According to the intensity of the whole gray mean, the edge detection is carried out by using heterogeneous operators and combining with a variety of threshold filtering methods to extract the lane pixels. The lane detection module transmits the vehicle steering angle and the deviate distance from the road center line to the decision layer, effectively implementing the interactive simulation. Experiments show that the algorithm has good real-time performance, stability, and robustness under different illumination and road conditions.

Pearl gentian grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) is a hybrid fish with high commercial value. It is widely cultured on the Asian coast; however, it is not cold-tolerant. Although we have previously characterized the liver transcriptomic responses of this grouper to cold stress, the roles of miRNAs and transcription factors (TFs) in cold resistance and the underlying regulatory mechanisms are still unclear. In this study, we integrated miRNA and mRNA sequencing data for pearl gentian grouper under cold stress and constructed a miRNA-TF-mRNA regulatory network. Furthermore, we screened seven key miRNAs (i.e., gmo-miR-221-5p, ssa-miR-7132b-5p, ola-let-7c, ssa-miR-25-3-5p, ccr-miR-489, gmo-miR-10545-5p, ccr-miR-122) that regulated target genes (including TF ACSS2, TF PPARD, TF PPP4CB; CYP2J2, EHHADH, RXRs, NR1D2, PPP1CC-A, PPP2R1A, FOXK2, etc.). These miRNAs participated in several important pathways and biological processes by the direct or indirect regulation of target genes, such as antioxidation and membrane fluidity, glucose and lipid metabolism, circadian rhythm, DNA repair, and apoptosis. The key cold-related miRNAs, TFs, and genes and their potential regulatory relationships identified in this study provide a deeper understanding of the complex molecular basis of the response to low-temperature environments in the grouper. In particular, our results provide the first identification for the role of NR1D2 gene in the cold tolerance of fish via the regulation of circadian rhythm. Furthermore, the key miRNAs and genes provide a basis for the molecular breeding of new cold-tolerant varieties of the pearl gentian grouper.

Pigs have anatomical and physiological characteristics comparable to those in humans and, therefore, are a favorable model for immune function research. Interferons (IFNs) and inflammasomes have essential roles in the innate immune system. Here, we report that G10, a human-specific agonist of stimulator of interferon genes (STING), activates both type I IFN and the canonical NLRP3 inflammasome in a STING-dependent manner in porcine cells. Without a priming signal, G10 alone transcriptionally stimulated Sp1-dependent p65 expression, thus triggering activation of the nuclear factor-κB (NF-κB) signaling pathway and thereby priming inflammasome activation. G10 was also found to induce potassium efflux- and NLRP3/ASC/Caspase-1-dependent secretion of IL-1β and IL-18. Pharmacological and genetic inhibition of NLRP3 inflammasomes increased G10-induced type I IFN expression, thereby preventing virus infection, suggesting negative regulation of the NLRP3 inflammasome in the IFN response in the context of STING-mediated innate immune activation. Overall, our findings reveal a new mechanism through which G10 activates the NLRP3 inflammasome in porcine cells and provide new insights into STING-mediated innate immunity in pigs compared with humans.

Angular glints are induced essentially by the superposition of scattering responses from multiple scattering centers and usually fluctuate quickly and widely with the aspect angles for complex targets change. This paper presents angular glint simulation of complex conducting targets using attributed scattering centers. The formulae for linear glint errors (LGEs) are derived based on the parametric models of attributed scattering centers and can be applied for real-time LGE simulation. Our analyses and numerical experiments show that the LGE simulation requires more precise scattering center modeling than the radar imaging simulation does. To ensure the accuracy of the LGE simulation, a forward approach that uses induced currents on conducting targets is used to precisely extract the scattering centers. The numerical results show that the LGE simulation based on the developed scattering center model has high accuracy. More importantly, the scattering center model developed for sparse aspect angles can be used to simulate LGEs for dense aspect angles, as required for complex targets, which leads to high efficiency in simulating LGEs.

In this study, we determined the complete mitogenome of Nemipterus virgatus of which the length was 17,073 bp, including 37 canonical mitochondrial genes and 2 non-coding regions. The control region contained termination associated sequence domain (TAS), central conserved domain (CSB-F, CSB-E, CSB-D, CSB-C, and CSB-A), conserved sequence block domain (CSB-1, CSB-2, and CSB-3), and tandem repeat sequence domain (TTD). Nine single nucleotide polymorphisms and three insertion of tandem repeat sequence (each length in 28 bp) were detected between two N. virgatus mitogenomes. The phylogenetic analysis showed that the families Nemipteridae, Sparidae, Centracanthidae, and Lethrinidae did not gather into a monophyly of superfamily Sparoidea in the neighbor-joining tree.

In this study, we reported the complete mitogenome of Scolopsis ciliata sampled from the Beibu Gulf, South China Sea. It was 16,734 bp in length and contained 37 canonical mitochondrial genes and 2 non-coding regions. Fifty-one single nucleotide polymorphisms and five insertion and deletions were detected between two S. ciliata mitogenomes, suggesting a high level of intraspecific genetic variation in the species. The control region contained termination associated sequence domain (TAS), central conserved domain (CSB-F, CSB-E, CSB-D, CSB-C, and CSB-A), and conserved sequence block domain (CSB-1, CSB-2, and CSB-3). The phylogenetic analysis of 21 complete mitogenome sequences well supports the phylogenetic position of S. ciliata and reveals the phylogenetic relationship of the genus Scolopsis.

The complete mitochondrial genome of the ornate threadfin bream, Nemipterus hexodon, was first determined by the pairs-walking sequencing in this study. The circular mtDNA molecule was 17,115 bp in size and the overall nucleotide composition of H-stand was A (29.55%), T (27.36%), G (16.08%), and C (27.01%), with a slight bias towards A + T. The complete mitogenome encoded 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and 2 non-coding regions (an origin of L-strand replication and a control region). The Bayesian tree supported the phylogenetic position of N. hexodon, which provided useful information for phylogenetic relationship in genus Nemipterus.

Pearl gentian grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) is a fish of high commercial value in the aquaculture industry in Asia. However, this hybrid fish is not cold-tolerant, and its molecular regulation mechanism underlying cold stress remains largely elusive. This study thus investigated the liver transcriptomic responses of pearl gentian grouper by comparing the gene expression of cold stress groups (20, 15, 12, and 12 °C for 6 h) with that of control group (25 °C) using PacBio SMRT-Seq and Illumina RNA-Seq technologies. In SMRT-Seq analysis, a total of 11,033 full-length transcripts were generated and used as reference sequences for further RNA-Seq analysis. In RNA-Seq analysis, 3271 differentially expressed genes (DEGs), two low-temperature specific modules (tan and blue modules), and two significantly expressed gene sets (profiles 0 and 19) were screened by differential expression analysis, weighted gene co-expression networks analysis (WGCNA), and short time-series expression miner (STEM), respectively. The intersection of the above analyses further revealed some key genes, such as PCK, ALDOB, FBP, G6pC, CPT1A, PPARα, SOCS3, PPP1CC, CYP2J, HMGCR, CDKN1B, and GADD45Bc. These genes were significantly enriched in carbohydrate metabolism, lipid metabolism, signal transduction, and endocrine system pathways. All these pathways were linked to biological functions relevant to cold adaptation, such as energy metabolism, stress-induced cell membrane changes, and transduction of stress signals. Taken together, our study explores an overall and complex regulation network of the functional genes in the liver of pearl gentian grouper, which could benefit the species in preventing damage caused by cold stress.

Late Pleistocene climate oscillations are believed to have greatly influenced the distribution, population dynamics, and genetic variation of many marine organisms in the western Pacific. However, the impact of the late Pleistocene climate cycles on the demographic history and population genetics of pelagic fish in the northern South China Sea (SCS) remains largely unexplored. In this study, we explored the demographic history, genetic structure, and genetic diversity of Decapterus maruadsi, a typical pelagic fish, over most of its range in the northern SCS. A 828–832 bp fragment of mitochondrial control region were sequenced in 241 individuals from 11 locations. High haplotype diversity (0.905–0.980) and low nucleotide diversity (0.00269–0.00849) was detected, revealing low levels of genetic diversity. Demographic history analysis revealed a pattern of decline and subsequent rapid growth in the effective population size during deglaciation, which showed that D. maruadsi experienced recent demographic expansion after a period of low effective population size. Genetic diversity, genetic structure, and phylogenetic relationship analysis all demonstrated that no significant genetic differentiation existed among the populations, indicating that D. maruadsi was panmictic throughout the northern SCS. Periodic sea-level changes, fluctuation of the East Asian Monsoon, and Kuroshio variability were responsible for the population decline and expansion of D. maruadsi. The demographic history was the primary reason for the low levels of genetic diversity and the lack of significant genetic structure. The life history characteristics and ocean currents also had a strong correlation with the genetic homogeneity of D. maruadsi. However, the genetic structure of the population (genetic homogeneity) is inconsistent with biological characteristics (significant difference), which is an important reminder to identify and manage the D. maruadsi population carefully.

This genome-wide association study on persistent hepatitis B virus (HBV) infection among Chinese confirms previously associated genetic loci while discovering a novel protective locus at 8p21.3. The study also demonstrates the nearby gene INST10 suppresses HBV replication in vitro.

By analyzing the characteristics of expansive soil from Pingdingshan, China, the shear strength parameters at different water contents, dry densities, and dry-wet cycles of expansive soil are obtained. It is found that, at higher soil-water content, the internal friction angle is 0° and the shallow layer of expansive soil slope will collapse and destroy; this has nothing to do with the height of the slope and the size of the slope. The parameters of soil influenced by atmosphere are the ones which have gone through dry-wet cycles, and the parameters of soil without atmospheric influence are the same as those of natural soil. In the analysis of slope stability, the shear strength parameters of soil can be determined by using the finite element method, and the stability coefficient of the expansive soil slope can be calculated.

The complete mitogenome sequence of Dentex hypselosomus was amplified by designing 15 primer pairs. The circle genome was 16,618 bp in size and the overall base composition was 27.04% of A, 26.38% of T, 17.08% of G, and 29.50 of C, with significant anti-G bias. The complete mitogenome of D. hypselosomus encodes 37 canonical mitochondrial genes, two non-coding regions, an L-strand replication region (OL), and a control region (D-loop). The D-loop contained termination sequence domain (TAS), central conserved domains (CSB-F, CSB-E, CSB-D, CSB-C, and CSB-A), and conserved sequence blocks (CSB-1, CSB-2, and CSB-3). Phylogenetic analysis of nine sparid species well supported the phylogenetic position of D. hypselosomus and revealed the phylogenetic relationship of the family Sparidae at the level of mitochondrial genomes.