Establishment and characterization of a golden pompano (Trachinotus blochii) fin cell line for applications in marine fish pathogen immunology.

Pompano fishes have been widely farmed worldwide. As a representative commercial marine species of the Carangidae family, the golden pompano (Trachinotus blochii) has gained significant popularity in China and worldwide. However, because of rapid growth and high-density aquaculture, the golden pompano has become seriously threatened by various diseases. Cell lines are the most cost-effective resource for in vitro studies and are widely used for physiological and pathological research owing to their accessibility and convenience. In this study, we established a novel immortal cell line, GPF (Golden pompano fin cells). GPF has been passaged over 69 generations for 10 months. The morphology, adhesion and extension processes of GPF were evaluated using light and electron microscopy. GPF cells were passaged every 3 days with L-15 containing 20 % fetal bovine serum (FBS) at 1:3. The optimum conditions for GPF growth were 28 °C and a 20 % FBS concentration. DNA sequencing of 18S rRNA and mitochondrial 16S rRNA confirmed that GPF was derived from the golden pompano. Chromosomal analysis revealed that the number pattern of GPF was 48 chromosomes. Transfection experiments demonstrated that GPF could be utilized to express foreign genes. Furthermore, heavy metals (Cd, Cu, and Fe) exhibited dose-dependent cytotoxicity against GPF. After polyinosinic-polycytidylic acid (poly I:C) treatment, transcription of the retinoic acid-inducible gene I-like receptor (RLR) pathway genes, including mda5, mita, tbk1, irf3, and irf7 increased, inducing the expression of interferon (IFN) and anti-viral proteins in GPF cells. In addition, lipopolysaccharide (LPS) stimulation up-regulated the expression of inflammation-related factors, including myd88, irak1, nfκb, il1ß, il6, and cxcl10 expression. To the best of our knowledge, this is the first study on the immune response signaling pathways of the golden pompano using an established fin cell line. In this study, we describe a preliminary investigation of the GPF cell line immune response to poly I:C and LPS, and provide a more rapid and efficient experimental material for research on marine fish immunology.

Molecular and functional characterization of golden pompano (Trachinotus blochii) TBK1 on IFN regulation.

The golden pompano (Trachinotus blochii), a pivotal commercial marine species in China, has gained significant popularity worldwide. However, accompanied with rapid growth and high density aquaculture, golden pompano has been seriously threatened by Nervous necrosis virus (NNV), while its molecular biology research regarding the innate immune system remains unexplored, which is crucial for understanding the activation of interferon (IFN) production and antiviral responses. In this study, we aimed to identify the characterization and function of golden pompano TANK-binding kinase 1 (gpTBK1), thereby providing evidence of the conservation of this classical factor in the RLR pathway among marine fish. Initially, we found the expression of gpTBK1 upregulation in diseased golden pompano with NNV infection and we successfully cloned the full-length open reading frame (ORF) of gpTBK1, consisting of 2172 nucleotides encoding 723 amino acids, from the head kidney. Subsequent analysis of the amino acid sequence revealed homology between gpTBK1 and other fish TBK1 proteins, with conserved N-terminal Serine/Threonine protein kinases catalytic domain (S_TKc) and C-terminal coiled coil domain (CCD). Moreover, the expression pattern showed that gpTBK1 exhibited ubiquitous expression across all evaluated tissues. Furthermore, functional identification experiments indicated that gpTBK1 activated interferon promoters' activity in golden pompano and induced the expression of downstream IFN-stimulated genes (ISGs). Notably, gpTBK1 was found to co-localize and interact with gpIRF3 in the cytoplasm. Collectively, these data provide a comprehensive analysis of the characterization and functional role of gpTBK1 in promoting interferon production. This research may facilitate the further study of the innate antiviral response, particularly the anti-NNV mechanisms, in golden pompano.

Transcriptome analysis reveals candidate miRNAs involved in skin color differentiation of juvenile Plectropomus leopardus in response to different background colors.

Red skin color in Plectropomus leopardus is important to its ornamental and economic value. However, the color of P. leopardus can change during the rearing process, darkening and turning black due to the influence of environmental background color. The underlying molecular mechanisms that regulate this phenomenon remain unclear. MicroRNAs (miRNAs) are endogenous, small non-coding RNAs that play important roles in numerous biological processes, such as skin differentiation and color formation in many animals. Therefore, we performed miRNA sequencing of P. leopardus skin before (initial) and after rearing with three different background colors (white, black, and blue) using Illumina sequencing to identify candidate miRNAs that may contribute to skin color differentiation. In total, 154,271,376 clean reads were obtained, with over 92 % of them successfully mapped to the P. leopardus reference genome. The miRNA length distributions of all samples displayed peaks around a typical length of 22 nt. Within these sequences, 243 known and 287 novel miRNAs were identified. A total of 65 significantly differentially expressed miRNAs (DEMs) were identified (P < 0.05), including 40 known DEMs and 25 novel DEMs. These DEMs included novel_561, miR-141-3p, and miR-129-5p, whose target genes were primarily associated with pigmentation related processes, including tyrosine metabolism, melanogenesis, and the Wnt signaling pathway. These findings shed light on the potential roles of miRNAs in the darkening of skin color in P. leopardus, thus enhancing our understanding of the molecular mechanisms involved in skin pigmentation differentiation in this species.

The muscle nutritional components analysis of golden pompano (Trachinotus blochii) in different mariculture area, growth stages, and genders.

Golden pompano (Trachinotus blochii) is an economically important fish which exhibits sexual size dimorphism and is widely cultivated in the southern seas of China. To evaluate the nutritional composition of T. blochii of different mariculture areas, growth stages, and genders, the moisture, ash, amino acids, and fatty acids in the muscle were measured using national standard biochemical assay. The analysis found 16 kinds of amino acids in the muscle of T. blochii. The EAA contents of fish from Guangdong (GD) and Guangxi (GX) were significantly lower than those of Hainan (HN) and Fujian (FJ) (p < 0.05). The unsaturated fatty acids were higher in T. blochii cultured in HN and FJ (p < 0.05). Within the same sea area, the contents of TAA, EAA, DAA, and PUFA increased with growth in T. blochii, but the differences were not significant (p > 0.05). EAA/TAA and EAA/NEAA conformed to the ideal FAO/WHO model. The AAS, CS, and EAAI scores of amino acids within groups gradually increased with growth. The TAA, EAA and PUFA contents in females were higher than in males (p > 0.05). The slightly higher amounts of amino acids and fatty acids in female T. blochii indicated females had higher nutritional value. In conclusion, the HN and FJ groups, the later growth stages, and the female T. blochii had generally higher nutritional values than their respective counterparts. These results provide fundamental data supporting all-female T. blochii breeding and culture, and optimized marketing body size.

Weighted correlation network analysis of the genes in the eyes of juvenile Plectropomus leopardus provide novel insights into the molecular mechanisms of the adaptation to the background color.

Plectropomus leopardus is a valuable marine fish whose skin color is strongly affected by the background color. However, the influence of the visual sense on the skin color variation of P. leopardus remains unknown. In the present study, transcriptome analysis was used to examine the visual response mechanism under different background colors. Paraffin sections of the eyes showed that the background color caused morphological changes in the pigment cells (PCs) and outer nuclear layer (ONL) and the darkening of the iris color. The transcriptome analysis results indicated that the gene expressions in the eyes of P. leopardus were significantly different for different background colors. We identified 4845, 3069, 5874, and 6309 differentially expressed genes (DEGs) in the pairwise comparisons of white vs. initial, blue vs. initial, red vs. initial, and black vs. initial groups, respectively. Some hub genes and key pathways regulating the adaptive mechanism of P. leopardus's eyes to the background color were identified, i.e., the JAK-STAT, mTOR, and Ras signaling pathways, and the ndufb7, slc6a13, and novel.3553 gene. This adaptation was achieved through the synthesis of stress proteins and energy balance supply mediated by hub genes and key pathways. In addition, the phenylalanine metabolism, tyrosine metabolism, and actin cytoskeleton-related processes or pathways and genes were responsible for iris and skin color adaptation. In summary, we inferred that stress protein synthesis, phenylalanine metabolism, and energy homeostasis were critical stress pathways for P. leopardus to adapt its skin color to the environment. These new findings indicate that the P. leopardus skin color variation may have been caused by the environmental adaption of the eyes. The results provide new insights into the molecular mechanisms underlying the skin color adaptation of P. leopardus.

A protocol for gradient coating design zinc anodes for stable zinc ion batteries.

In this protocol, we present a modified gradient coating strategy for zinc anodes. We describe steps for synthesizing electrodes, measuring electrochemistry, and assembling and testing batteries. The protocol can be applied for broadening design ideas of functional interface coating. For complete details on the use and execution of this protocol, please refer to Chen et al. (2023).1.

Reactive oxygen species-mediated phosphorylation of JNK is involved in the regulation of BmFerHCH on Bombyx mori nucleopolyhedrovirus proliferation.

c-Jun N-terminal kinase (JNK) phosphorylation is widely observed during virus infection, modulating various aspects of the virus-host interaction. In our previous research, we have proved that B. mori ferritin heavy-chain homolog (BmFerHCH), an inhibitor of reactive oxygen species (ROS), facilitates B. mori nucleopolyhedrovirus (BmNPV) proliferation. However, one question remains: Which downstream signaling pathways does BmFerHCH regulate by inhibiting ROS? Here, we first determined that silencing BmFerHCH inhibits BmNPV proliferation, and this inhibition depends on ROS. Then, we substantiated that BmNPV infection activates the JNK signaling pathway. Interestingly, the JNK phosphorylation during BmNPV infection is activated by ROS. Further, we found that the enhanced nuclear translocation of phospho-JNK induced by BmNPV infection was dramatically reduced by pretreatment with the antioxidant N-acetylcysteine (NAC), whereas there was more detectable phospho-JNK in the cytoplasm. Next, we investigated how changes in BmFerHCH expression affect JNK phosphorylation. BmFerHCH overexpression suppressed the phosphorylation of JNK and nuclear translocation of phospho-JNK during BmNPV infection, whereas BmFerHCH knockdown facilitated phosphorylation of JNK and nuclear translocation of phospho-JNK. By measuring the viral load, we found the inhibitory effect of BmFerHCH knockdown on BmNPV infection depends on phosphorylated JNK. In addition, the JNK signaling pathway was involved in BmNPV-triggered apoptosis. Hence, we hypothesize that ROS-mediated JNK phosphorylation is involved in the regulation of BmFerHCH on BmNPV proliferation. These results elucidate the molecular mechanisms and signaling pathways of BmFerHCH-mediated response to BmNPV infection.

Transcriptome analysis of differentially expressed circular RNAs in the testis and ovary of golden pompano (Trachinotus blochii).

The artificial breeding of golden pompano (Trachinotus blochii) has expanded greatly in recent years, and after long-term breeding efforts, clear sexual dimorphisms have been observed in T. blochii growth traits, with females growing faster. As sponges of microRNA (miRNAs), circular RNAs (CircRNAs) can alleviate miRNA inhibition of target mRNA. However, few studies have examined sex-related CircRNAs and none of those have looked at T. blochii. To further understand the role of CircRNAs in sex differentiation and sexual size dimorphism in T. blochii, six CircRNA libraries were constructed from the testes and ovaries of T. blochii. A total of 1522 CircRNAs were found distributed over all 24 chromosomes of T. blochii. 135 differentially expressed CircRNAs (DECs) were identified by screening, These DECs were then subjected to GO enrichment, which found 47 enriched pathways. A number of CircRNAs were enriched in cellular processes and metabolic processes. According to the KEGG pathway analysis, a series of sex differentiation pathways were enriched, including the GnRH, calcium, and MAPK signaling pathways. Furthermore, we selected two CircRNAs from the DECs named circ-cacna1b and circ-octc. We found that the cacna1b gene is regulated by 7 miRNAs, 3 of which were regulated by circ-cacna1b, i.e., mmu-miR-138-5p, fru-miR-138, and pma-miR-138b. In addition, the miRNA named pma-miR-138b can regulate sex-related genes, such as sox9 and dmrt1, among others. The co-expression network of CircRNA-miRNA-mRNA showed circ-cacna1b may play a crucial role in T. blochii sex differentiation by regulating pma-miR-138b to affect the expression of sex differentiation genes. The circ-octc may be one of the largest contributors to sexual size dimorphism during growth through its effect on lipid metabolism. These findings could broaden our understanding of CircRNAs and provide new insight into their function in sex differentiation and growth.

Identification and Characterization of Sex-Biased miRNAs in the Golden Pompano (Trachinotus blochii).

The golden pompano (Trachinotus blochii) is a marine fish of considerable commercial importance in China. It shows notable sexual size dimorphism; the growth rate of females is faster than that of males. Therefore, sex-biased research is of great importance in T. blochii breeding. However, there have been few studies on sex differentiation and mechanisms underlying sex determination in T. blochii. MicroRNAs (miRNAs) play crucial roles in sex differentiation and determination in animals. However, limited miRNA data are available on fish. In this study, two small RNA libraries prepared from the gonads of T. blochii were constructed and sequenced. The RNA-seq analysis yielded 1366 known and 69 novel miRNAs with 289 significantly differentially expressed miRNAs (p < 0.05). Gene ontology (GO) analysis confirmed that the TFIIA transcription factor complex (GO: 0005672) was the most significantly enriched GO term. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the differentially expressed miRNAs and target genes were mainly related to sex determination and gonadal developmental signaling pathways, specifically the Wnt signaling pathway, MAPK signaling pathway, and steroid biosynthetic pathway. MiRNA-mRNA co-expression network analysis strongly suggested a role for sex-biased miRNAs in sex determination/differentiation and gonadal development. For example, gata4, foxo3, wt1, and sf1 genes were found to be regulated by bta-miR-2898; esr2 and foxo3 by novel_176, and ar by oar-let-7b. Quantitative real-time polymerase chain reaction analysis of selected mRNAs and miRNAs validated the integrated analysis. This study established a set of sex-biased miRNAs that are potential regulatory factors in gonadal development in T. blochii. These results provide new insight into the function of miRNAs in sex differentiation and determination in T. blochii and highlight some key miRNAs for future studies.

The Effect of Background Color on Skin Color Variation of Juvenile Plectropomus leopardus.

Fish skin color is usually strongly affected by the background color of their environment. The study investigated the effects of five different background colors on the skin color of leopard coral groupers (Plectropomus leopardus). More than 450 juveniles were reared in Blue, Red, Black, White, and Transparent background tanks for 56 days. The paraffin section showed that the skin melanin zone of fish in the White group was smaller, whereas the Black and Red groups (especially Black) were nearly the largest. The apparent skin color of P. leopardus was red on the white background, which darkened in response to the other color backgrounds. The Black group revealed the blackest skin color, followed by the transparent group. Moreover, the White group had the highest L*, a*, and b* values. The melanin content and tyrosinase activity in the dorsal and ventral skin of the Black group were significantly higher than those in the other groups (p < 0.05), and the serum α-MSH level was higher in the Black group as well. The carotenoid and lutein contents showed completely different trends among the experimental groups, as carotenoid content was higher in the Red and White groups, while lutein content was higher in the Transparent group. The expression level of scarb1 was highest in the Blue and White groups, followed by the Transparent group, and lowest in the Black group (p < 0.05). The expression trend of scarb1 was similar to the skin color in different backgrounds, indicating that the background color regulated scarb1 expression level through visual center, then influenced the uptake and transport of carotenoids, then influenced the skin color formation of P. leopardus. Moreover, lighter colors inhibited the formation of melanocytes and had a significant effect on carotenoid and lutein contents. Pigment-related genes were involved in the regulation of fish skin color, and they were affected by background color in P. leopardus. These results indicate that a white background is more conducive to maintaining red skin color in juvenile P. leopardus.

Transcriptome Analysis Reveals the Complex Regulatory Pathway of Background Color in Juvenile Plectropomus leopardus Skin Color Variation.

Fish skin color is often strongly affected by background color. We hypothesized that the regulatory mechanism of variations in skin color in P. leopardus is linked to the background color. In this study, we conducted transcriptome analysis of Plectropomus leopardus cultured under different background colors to compare gene expression levels and the important signaling pathways. The RNA-seq analysis yielded 26,675 known mRNAs, 3278 novel mRNAs, and 3179 differentially expressed genes (DEGs). The DEGs related to melanin synthesis were screened out. Some key melanin-related genes were identified, specifically tyr, slc7a11, mc1r, ednrb, dct, tat, and wnt1. These DEGs were mainly involved in melanogenesis, including tyrosine metabolism, the Wnt signaling pathway, and the cAMP signaling pathway. The expression levels of some key genes were upregulated when background color deepened, such as α-msh, wnt, and gf. The α-MSH/cAMP-dependent, Wnt/ß-catenin, and PI3K/Akt signaling pathways were activated, resulting in the accumulation of intracellular mitf. mitf promoted melanin production by binding to the tyr/tyrp1/dct promoter region. In the present study, we explored the molecular mechanism underlying the darkened skin color pattern of P. leopardus, providing a theoretical basis for the molecular mechanism underlying pigmentation in P. leopardus.

Mesoporous vanadium nitride as anion storage electrode for reverse dual-ion hybrid supercapacitor.

In traditional dual-ion systems, the cathode usually is employed as anion-storage materials. Herein, we propose a new dual-ion hybrid supercapacitor with reverse anion/cation-storage mechanism, consisting of a mesoporous (MPs) VN anode as a pivotal anion-storage material and K2-xMn8O16 nanosheet arrays grown on carbon cloth (NSs/CC) as (K-storage) cathode. During charge/discharge, the anode and cathode reversibly store/release OH- ions and K+ ions, respectively. Herein, the MPs VN as anion-storage electrode can operate in an alkaline condition and deliver a high capacitance of 251 mF cm-2 with desired low-voltage plateau. More importantly, benefiting from unique reverse dual-ion mechanism, the (MPs VN-K2-xMn8O16 NSs/CC) hybrid device displays excellent rate performance and satisfying area capacitance along with good durability of 92.2% after 10,000 cycles at a scan rate of 100 mV s-1. It offers new ideas to expand the range of anion-storage materials in dual-ion hybrid supercapacitors.

Characterization and functional analysis of myostatin and myogenin genes involved in temperature variation and starvation stress in Golden pompano, Trachinotus blochii.

Animal growth and development is a complicated process and is regulated by multi-genes. Myostatin (Mstn) and myogenin (Myog) are a pair of negative and positive regulators respectively, which play an important role in the generation of muscle cells. In order to study the function of these two genes in muscle growth of Trachinotus blochii, full lengths of two mstn genes (mstn-1 and mstn-2) and myog gene were cloned using RACE. We first identified and characterized the complete cDNA sequences of mstn-1, mstn-2, and myog genes derived from T. blochii, an economically important mariculture species in China. Multiple sequence alignment of amino acids and phylogenetic analysis revealed that the Mstn and Myog were highly conserved to the other Perciformes. In addition, gene duplication of mstn in T. blochii was observed. mstn-1 mRNA was mainly expressed in the muscle and gonad, while mstn-2 and myog transcripts were detectable mainly in the brain and muscle, respectively. Moreover, the nutritional status and temperature influenced abundance levels in brain and muscle. Results suggested that mstn and myog genes play an important role in muscle growth of T. blochii, mstn may not be limited to control of muscle growth in fish and could also be involved in other biological functions.

Multifunctional integrated VN/V2O5 heterostructure sulfur hosts for advanced lithium-sulfur batteries.

Lithium-sulfur (Li-S) batteries show great potential in future electric transportation and large-scale grid storage applications because of their attractive theoretical energy density (2600 W h kg-1) and relatively abundant sulfur reserves. However, the rapid capacity decay and unsatisfactory sulfur loading caused by the lithium polysulphide (LiPS) dissolution and low electrical conductivity of sulfur are the most urgent issues plaguing its practical applications. Herein, we report a multifunctional nanoporous (NP) VN/V2O5 binary host that can efficiently resolve the above conflicts by the synergy between the functions of two materials. The inner V2O5 facilitates rapid trapping of numerous LiPSs while the outer porous VN with abundant NP channels offers high conductivity and mild chemisorption, thereby improving the localization and catalytic conversion ability of LiPSs. Accordingly, the designed cathodes with 1.87 mg cm-2 sulfur content achieve an acceptable areal specific capacity (2.72 mA h cm-2), excellent rate capability (963 mA h g-1 at 5.0C), and cycling stability. Remarkably, the cathodes with ultrahigh sulfur loading of 9.02 mg cm-2 deliver a satisfactory areal specific capacity (12.12 mA h cm-2) and still maintain excellent durability.

A new potassium dual-ion hybrid supercapacitor based on battery-type Ni(OH)2 nanotube arrays and pseudocapacitor-type V2O5-anchored carbon nanotubes electrodes.

Hybrid supercapacitors (HSCs) with the characteristics of high energy density, long cycle life and without altering their power density need to be developed urgently. Herein, a novel dual-ion hybrid supercapacitors (DHSCs) with Ni(OH)2 nanotube arrays (NTAs) as positive electrode and V2O5 directly grown on freestanding carbon nanotubes (CNTs) as negative electrode is assembled. In charging mechanism of DHSCs, K+ are inserted into the V2O5 negative while OH- react with Ni(OH)2 positive; during discharge, the K+ and OH- are released from V2O5 negative and Ni(OH)2 positive, respectively, and return back to the electrolyte, which is quite different from traditional metal ion or alkaline supercapacitors. Because of the merits combining dual-ion mechanism and HSCs, the DHSC displays excellent capacity retention of âˆ¼ 81.4% after 10,000 cycles, high energy density of âˆ¼ 25.4 µWh cm-2 and high power density of âˆ¼ 4.66 mW cm-2, indicating the potential applications in the further on flexible wearable electronics.

Identification of Candidate Genes Associated With Hypoxia Tolerance in Trachinotus blochii Using Bulked Segregant Analysis and RNA-Seq.

Golden Pompano (Trachinotus blochii) has rapidly developed into the one of the main valuable fish species in Chinese marine aquaculture. Due to its rapid growth, active metabolism, and high oxygen consumption, hypoxia will increase its mortality and cause serious economic losses. We constructed two experimental groups of fish with different degrees of tolerance to hypoxia, used BSR-Seq analysis based on genome and genetic linkage groups to locate SNPs and genes that were related to the differences in hypoxia tolerance. The results showed that hypoxia tolerance SNPs of golden pompano may be jointly determined by multiple linkage groups, especially linkage groups 18 and 22. There were 768 and 348 candidate genes located in the candidate regions of the brain and liver, respectively. These genes were mainly involved in anaerobic energy metabolism, stress response, immune response, waste discharge, and cell death. The prostaglandin-endoperoxide synthase 2 (PTGS2) on LG8, which is involved in the metabolism of arachidonic acid, has a G/A nonsynonymous mutation at position 20641628, and the encoded amino acid was changed from hydrophobic aspartic acid to asparaginate. The specific pathway of the RIG-I-like receptor signaling pathway in the liver may mediate the metabolic system and the immune system, linking glucose metabolism with immune regulation. The death of the hypoxia-intolerant group may be due to the accumulation of lactic acid caused by the activation of anaerobic glycolysis during the early stage of hypoxia stress, and the activation of type I interferon was inhibited, which resulted in decreased immunity. Among the genes involved in the RIG-I-like receptor signaling pathway, the CYLD Lysine 63 Deubiquitinase (CYLD) located on LG16 had a G/T nonsynonymous mutation at position 13629651, and the encoded amino acid was changed from alanine acid to valine. The interferon induced with helicase C domain 1 (Ifih1) located on LG18 has a G/C nonsynonymous mutation at position 16153700, and the encoded hydrophilic glycine was changed to hydrophobic alanine. Our findings suggest these SNPs may assist in the molecular breeding of hypoxia-tolerant golden pompano, and speculate that the balance of glucose and lipid metabolism plays a key role in Trachinotus blochii under acute hypoxia.

EZH2 is a potential prognostic predictor of glioma.

The enhancer of zeste homologue 2 (EZH2) is a histone H3 lysine 27 methyltransferase that promotes tumorigenesis in a variety of human malignancies by altering the expression of tumour suppressor genes. To evaluate the prognostic value of EZH2 in glioma, we analysed gene expression data and corresponding clinicopathological information from the Chinese Glioma Genome Atlas, the Cancer Genome Atlas and GTEx. Increased expression of EZH2 was significantly associated with clinicopathologic characteristics and overall survival as evaluated by univariate and multivariate Cox regression. Gene Set Enrichment Analysis revealed an association of EZH2 expression with the cell cycle, DNA replication, mismatch repair, p53 signalling and pyrimidine metabolism. We constructed a nomogram for prognosis prediction with EZH2, clinicopathologic variables and significantly correlated genes. EZH2 was demonstrated to be significantly associated with several immune checkpoints and tumour-infiltrating lymphocytes. Furthermore, the ESTIMATE and Timer Database scores indicated correlation of EZH2 expression with a more immunosuppressive microenvironment for glioblastoma than for low grade glioma. Overall, our study demonstrates that expression of EZH2 is a potential prognostic molecular marker of poor survival in glioma and identifies signalling pathways and immune checkpoints regulated by EHZ2, suggesting a direction for future application of immune therapy in glioma.

Co-modulation of Liver Genes and Intestinal Microbiome of Largemouth Bass Larvae (Micropterus salmoides) During Weaning.

In recent years, largemouth bass have become one of the most commonly aquacultured species in China, however, its low survival rate during larval weaning has always been a bottleneck that has restricted industrial development. Understanding the changes in liver metabolism and intestinal microflora during the weaning of largemouth bass larvae can help to design better weaning strategies and improve survival. In this study, liver mRNA and intestinal microflora 16S rRNA genes were analyzed using high-throughput sequencing at the pre, mid, and post weaning stages [15, 30, 45 days post hatching; total length (cm) were 2.21 ± 0.12, 3.45 ± 0.21, 5.29 ± 0.33, respectively]. The transcriptome results revealed that the genes with increased expression were related to amino acid metabolism in the pre-weaning stage, but they were related to fatty acid metabolism in the post-weaning stage. A similar phenomenon was observed in the intestinal microflora where the dominant microbe Proteobacteria (relative abundance 56.32%) in the pre-weaning stage was gradually replaced by Firmicutes (relative abundance 62.81%) by the post-weaning stage. In addition, the three most important digestive enzymes (trypsin, lipase, and amylase) in the intestine were significantly decreased during the mid-weaning stage (P < 0.05), which was also true for some genes crucial to immune pathways in the liver. Overall, these findings showed that weaning in largemouth bass can cause changes in liver metabolism and intestinal microbial communities, which has improved our understanding of fish adaptation to changes in food sources during weaning.

Combined exposure to hypoxia and ammonia aggravated biological effects on glucose metabolism, oxidative stress, inflammation and apoptosis in largemouth bass (Micropterus salmoides).

Hypoxia and ammonia are unavoidable environmental factors in aquaculture, and have been shown cause various adverse effects in fish. In the present study, a two-factor crossover experiment was carried out to evaluate the combined effect of hypoxia and ammonia on oxidative stress and glucose metabolism endpoints in largemouth bass. The fish were divided into four experimental groups: hypoxia and ammonia group, hypoxia group, ammonia group, and control group. The results showed that hypoxia and ammonia exposures both induced antioxidant response and oxidative stress (superoxide dismutase [SOD] and catalase [CAT] activities increased first then decreased, and malondialdehyde accumulated) and anaerobic glycolysis (increase of blood glucose, decrease of liver glycogen, accumulation of lactate, and increased lactate dehydrogenase activity). In addition, hypoxia and ammonia upregulated antioxidant enzyme genes (Cu/ZnSOD, CAT, and GPx), apoptosis genes (caspase 3, caspase 8, and caspase 9), as well as inflammatory genes (interleukin [IL]-1ß and IL-8) and downregulated an anti-inflammatory gene (IL-10), suggesting that apoptosis and inflammation may be related to oxidative stress. The increased expression of GLUT1, LDH, and MCT4 were induced by hypoxia and ammonia, suggesting that anaerobic glycolysis was increased. Furthermore, fish suffering from hypoxia or ammonia exposure showed some changes in gill tissues histology, and the most severe lesions of gill tissues appeared in simultaneous exposure. Overall, both hypoxia and ammonia affected homeostasis, and simultaneous exposure led to more deleterious effects on largemouth bass than exposure to the individual stressors.

Pathological Grade-Associated Transcriptome Profiling of lncRNAs and mRNAs in Gliomas.

The aim of the present study was to explore the expression profiles of lncRNAs and mRNAs in glioma patients and to elucidate any potential relationship between lncRNAs and mRNAs in glioma. High-throughput transcriptome sequencing of mRNAs and lncRNAs from six normal tissues and 16 glioma tissues (grade II, six cases; grade III, four cases; and grade IV, six cases) was performed. Series test of cluster (STC) analysis was used to screen significant trending models associated with glioma. Gene co-expression networks were constructed for the differentially expressed lncRNAs and mRNAs, and gene-ontology (GO) and pathway-enrichment analyses were further performed. Quantitative real-time PCR was performed to validate the five most differentially expressed lncRNAs and mRNAs. After filtering the raw sequencing data, we found 578 lncRNAs and 3,216 mRNAs that were significantly dysregulated in glioma (fold change ≥ 2, p < 0.05). Twenty model profiles of lncRNA and 10 model profiles of mRNA were summarized, and three patterns of lncRNAs and two patterns of mRNAs were of clinical significance. Three gene co-expression networks between mRNAs and lncRNAs were built to clarify the relationship between lncRNAs and mRNAs in glioma. GO and pathway analyses indicated that the differentially expressed lncRNAs and mRNAs were enriched in several biological processes and signaling pathways associated with tumorigenesis. Both lncRNAs and mRNAs exhibited dynamic differential expression profiles that indicated their potential roles in different degrees of glioma malignancy. A series of bioinformatics analyses indicated that most of these lncRNAs and mRNAs are involved in important biological processes and pathways associated with the pathogenesis of glioma. These results provide potential directions and valuable resources for future investigations via the comprehensive integration of these lncRNAs and mRNAs.