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Objective: To explore the correlation of formyl peptide receptor 1 expression with drug resistance and the functions of formyl peptide receptor 1 in drug-resistant bladder cancer., Methods: Expression of formyl peptide receptor 1 in T24 and T24/DDP cisplatin-resistant bladder cancer cell lines was tested by quantitative real-time Polymerase Chain Reaction and Western blotting. After incubation of T24/DDP with N-formyl-Met-Leu-Phe, the phosphor proteins were tested by Western blot analysis. We characterized the functions of formyl peptide receptor 1 in T24/DDP cells by assessing proliferation, migration, and changes of cell cycles., Results: Formyl peptide receptor 1 was expressed in both T24 and T24/DDP, and it was overexpressed in T24/DDP compared with T24. Formyl peptide receptor 1 activation promoted the expression of the messenger RNA of resistance-related proteins, such as multidrug resistance-associated protein 1 (MRP1) and lung resistance-related protein (LRP). The expression of 4 signal pathway proteins were upregulated: signal transducer and activator of transcription 3, Janus kinase 2, extracellular regulated protein kinases, and protein kinase B, while the expression of phosphatidylinositol 3-kinase was observed to be downregulated in drug-resistant bladder cancer cells. Formyl peptide receptor 1 activation also improved the expression of phospho-signal transducer and activator of transcription 3 and phospho-extracellular regulated protein kinases 1/2 and promoted the proliferation and migration of T24/DDP cells. In addition, formyl peptide receptor 1 inhibition led to the change in the cell cycle in T24/DDP., Conclusions: The overexpression of formyl peptide receptor 1 may be related to drug-resistant bladder cancer and promotes the deterioration of drug-resistant bladder cancer.

Chrysanthemum indicum L. capitulum is an enriched source of flavonoids with broad-ranging biological activities, mainly due to their anti-inflammatory, anti-cancer, immune regulation, anti-microbial activity, hepatoprotective, and neuroprotective effects. The O-methylation of various secondary metabolites has previously been demonstrated to be mainly catalyzed by S-adenosyl-L-methionine-dependent O-methyltransferase (OMT) proteins encoded by the OMT gene family. However, limited comprehensive study was published on the OMT gene family, especially the CCoAOMT subfamily, involved in the O-methylation of flavonoids in Chrysanthemum. Here, we analyzed the spatiotemporal expression patterns of C. indicum OMT genes in leaf and flower at different developmental stages. Transcriptome sequencing and qRT-PCR analysis showed that COMTs were mainly highly expressed in capitulum, especially in full bloom, while CCoAOMTs were mainly highly expressed in leaves. Correlation analysis of OMT gene expression and flavonoids accumulation revealed that four OMTs (CHR00029120, CHR00029783, CHR00077404, and CHR00078333) were putatively involved in most methylated flavonoids biosynthesis in the capitulum. Furthermore, we identified a true CCoAOMT enzyme, CiCCoAOMT1, and found that it catalyzed O-methylation of quercetin and luteolin at the 3′-OH position. In summary, this work provides an important theoretical basis for further research on the biological functions of OMTs in C. indicum. [ABSTRACT FROM AUTHOR]

Callose is an important polysaccharide composed of beta-1,3-glucans and is widely implicated in plant development and defense responses. Callose synthesis is mainly catalyzed by a family of callose synthases, also known as glucan synthase-like (GSL) enzymes. Despite the fact that GSL family genes were studied in a few plant species, their functional roles have not been fully understood in woody perennials. In this study, we identified total of 84 GSL genes in seven plant species and classified them into six phylogenetic clades. An evolutionary analysis revealed different modes of duplication driving the expansion of GSL family genes in monocot and dicot species, with strong purifying selection constraining the protein evolution. We further examined the gene structure, protein sequences, and physiochemical properties of 11 GSL enzymes in Prunus mume and observed strong sequence conservation within the functional domain of PmGSL proteins. However, the exon–intron distribution and protein motif composition are less conservative among PmGSL genes. With a promoter analysis, we detected abundant hormonal responsive cis-acting elements and we inferred the putative transcription factors regulating PmGSLs. To further understand the function of GSL family genes, we analyzed their expression patterns across different tissues, and during the process of floral bud development, pathogen infection, and hormonal responses in Prunus species and identified multiple GSL gene members possibly implicated in the callose deposition associated with bud dormancy cycling, pathogen infection, and hormone signaling. In summary, our study provides a comprehensive understanding of GSL family genes in Prunus species and has laid the foundation for future functional research of callose synthase genes in perennial trees. [ABSTRACT FROM AUTHOR]

Oxidative stress, as a characteristic of cellular aerobic metabolism, plays a crucial regulatory role in the development and metastasis of gastric cancer (GC). Long noncoding RNAs (lncRNAs) are important regulators in GC development. However, research on the prognostic patterns of oxidative stress-related lncRNAs (OSRLs) and their functions in the immune microenvironment is currently insufficient. We identified the OSRLs signature (DIP2A-IT1, DUXAP8, TP53TG1, SNHG5, AC091057.1, AL355001.1, ARRDC1-AS1, and COLCA1) from 185 oxidative stress-related genes in The Cancer Genome Atlas (TCGA) cohort via random survival forest and Cox analyses, and the results were subsequently validated in the Gene Expression Omnibus (GEO) dataset. The patients were divided into high- and low-risk groups by the risk score of the OSRLs signature. Longer overall survival was detected in the low-risk group than in the high-risk group in both the TCGA cohort (P < 0. 001, HR = 0.43, 95% CI 0.31–0.62) and the GEO cohort (P = 0.014, HR = 0.67, 95% CI 0.48–0.93). Next, multivariate Cox analysis identified that the risk model was an independent prognostic characteristic (HR > 1, P = 0.005), and time-dependent receiver operating characteristic (ROC) curve analysis and nomogram analysis were utilized to evaluate the predictive ability of the risk model. Next, gene set enrichment analysis revealed that the immune-related pathway, Wnt/ β -catenin signature, mammalian target of rapamycin complex 1 signature, and cytokine‒cytokine receptor interaction was enriched. High-risk patients were more responsive to CD200, TNFSF4, TNFSF9, and BTNL2 immune checkpoint blockade. The results of qRT‒PCR further proved the accuracy of our bioinformatic analysis. Overall, our study identified a novel OSRLs signature that can serve as a promising biomarker and prognostic indicator, which provides a personalized predictive approach for patient prognosis evaluation and treatment. [ABSTRACT FROM AUTHOR]

Comparative transcriptome analysis of fiber tissues between Gossypium barbadense and Gossypium hirsutum could reveal the molecular mechanisms underlying high-quality fiber formation and identify candidate genes for fiber quality improvement. In this study, 759 genes were found to be strongly upregulated at the elongation stage in G. barbadense, which showed four distinct expression patterns (I–IV). Among them, the 346 genes of group IV stood out in terms of the potential to promote fiber elongation, in which we finally identified 42 elongation-related candidate genes by comparative transcriptome analysis between G. barbadense and G. hirsutum. Subsequently, we overexpressed GbAAR3 and GbTWS1, two of the 42 candidate genes, in Arabidopsis plants and validated their roles in promoting cell elongation. At the secondary cell wall (SCW) biosynthesis stage, 2275 genes were upregulated and exhibited five different expression profiles (I–V) in G. barbadense. We highlighted the critical roles of the 647 genes of group IV in SCW biosynthesis and further picked out 48 SCW biosynthesis-related candidate genes by comparative transcriptome analysis. SNP molecular markers were then successfully developed to distinguish the SCW biosynthesis-related candidate genes from their G. hirsutum orthologs, and the genotyping and phenotyping of a BC3F5 population proved their potential in improving fiber strength and micronaire. Our results contribute to the better understanding of the fiber quality differences between G. barbadense and G. hirsutum and provide novel alternative genes for fiber quality improvement. [ABSTRACT FROM AUTHOR]

BGLU genes play crucial role in the defense of biotic and abiotic stresses, this study identified the members of the BGLU family in cotton via bioinformatics methods. The sequence features, conserved domain structure, chromosome location, promoter elements and other family features were analyzed, and the transcriptome data and qRT-PCR were used to analyze the pathogenic stress response pattern of BGLU family genes. The results showed a total of 53 BGLU genes were identified and divided into 6 sub-families according to the phylogenetic relationship in upland cotton, and some genes were specifically expressed under pathogen stress. qPCR analysis and RNAseq data showed that GhBGLU5, GhBGLUU, GhBGLU18, GhBGLU26 and GhBGLU34 maintained higher expression level in resistant and susceptible cottons, implying that these genes positively regulated cotton Verticillium wilt resistance. This study provided theoretical basis for further analyzing the functions and molecular mechanisms of the BGLU genes in cotton. [ABSTRACT FROM AUTHOR]

The pharyngeal sac is a comparatively rare organ in the digestive tract among teleost fishes. However, our understanding of this remarkable organ in the silver pomfret (Pampus argenteus) is limited. In the present study, we examined the various morphological and histological characteristics of the pharyngeal sac using histochemical techniques and electron microscopy. The pharyngeal sac showed unique characteristics such as well-developed muscular walls, weakly keratinized epithelium, numerous goblet cells, and needle-like processes on the papillae. The porous cavity of the papillae contained numerous adipocytes and was tightly enveloped by type I collagen fibers. These structures might provide mechanical protection and excellent biomechanical properties for grinding and shredding prey. A comparison of gene expression levels between the pharyngeal sac and esophagus using RNA-seq showed that phenotype-associated genes (epithelial genes and muscle genes) were upregulated, whereas genes related to nutrient digestion and absorption were downregulated in the pharyngeal sac. These results support the role of the pharyngeal sac in shredding and predigesting food. Overall, these findings provide a clearer understanding of the pharyngeal sac morphology and explain the morphological adaptations of the digestive tract for feeding on gelatinous prey. To our knowledge, this is the first report on pharyngeal sac gene expression in P. argenteus. [ABSTRACT FROM AUTHOR]

As a micronutrient, vitamin C plays an important role in growth and immunity of fish, but the effects of vitamin C on Pampus argenteus were still unclear. Here, we added vitamin C with different concentrations (0, 350, 1050, 2100 and 4200 mg/kg) to the basic diet, and the optimal concentration of vitamin C was screened out as 1050 mg/kg through the growth indicators, antioxidant enzyme activities (SOD, CAT, GSH‐PX and MDA) and the expression of four immune‐related genes (IL‐2, IL‐3, IL‐17 and IL‐18). After feeding dietary vitamin C supplementation with optimal concentration for 4 weeks, 80 ml/t formaldehyde was used to damage the immune system of fish. Then, the activities of the antioxidant enzymes and immune‐related genes were tested to find out the protective effects of vitamin C on the immune system. The results showed that the activity of CAT, SOD and GSH‐PX increased, while the MDA content decreased. At the molecular level, the expression of IL‐2 and IL‐3 genes increased, but the expression of IL‐17 and IL‐18 genes was inhibited. These findings suggested that the dietary vitamin C supplementation improved fish growth, alleviated the stress reaction caused by formaldehyde treatment and improved the immunity of P. argenteus. [ABSTRACT FROM AUTHOR]

Objective. To investigate the expression of ATPase family AAA domain-containing protein 2 (ATAD2) and kinesin family member 4A (KIF4A) in esophageal squamous cell carcinoma (ESCC) tissues and their association with clinicopathological features and to explore the role of ATAD2 in regulating KIF4A expression and biological functions in ESCC cells and the effect of aspirin on their expression. Methods. The mRNA and protein expression of ATAD2 and KIF4A in the tissues of patients with ESCC were measured by RT-qPCR and immunohistochemistry, and the correlation between the expression of mRNA and clinicopathological characteristics was analyzed. Western blot and RT-qPCR were used to detect the interference efficiency and KIF4A expression after si-ATAD2 transfection in EC109 and KYSE30 cells. CCK-8 and Transwell assay were performed to investigate the effects of ATAD2 and aspirin on proliferation, migration, and invasion of ESCC cells. The effect of aspirin on the expression of ATAD2 and KIF4A in ESCC cells was measured by RT-qPCR and Western blot. Results. The expression of ATAD2 and KIF4A was upregulated in ESCC tissues, and both were correlated with the differentiation grades and lymph node metastasis. Knockdown of ATAD2 in ESCC cells significantly inhibited cell proliferation, migration, and invasion. Compared to the negative control group, the proliferation, migration, and invasion ability of ESCC cells in the aspirin-treated groups were decreased, and the expression of ATAD2 and KIF4A in ESCC cells was decreased after treating with aspirin for 48 h. Conclusion. The expression levels of ATAD2 and KIF4A are elevated in ESCC. ATAD2 promotes proliferation, migration, and invasion of ESCC cells by regulating KIF4A. Aspirin can inhibit the malignant behavior of ESCC cells by downregulating ATAD2 and KIF4A. [ABSTRACT FROM AUTHOR]

Background: Many studies about depression have focused on the dysfunctional synaptic signaling in the hippocampus that drives the pathophysiology of depression. Radix Bupleuri has been used in China for over 2000 years to regulate liver-qi. Extracted from Radix Bupleuri, Saikosaponin D (SSD) is a pharmacologically active substance that has antidepressant effects. However, its underlying mechanism remains unknown. Materials and methods: A chronic unpredictable mild stress (CUMS) paradigm was used as a rat model of depression. SD rats were randomly assigned to a normal control (NC) group or one exposed to a CUMS paradigm. Of the latter group, rats were assigned to four subgroups: no treatment (CUMS), fluoxetine-treated (FLU), high-dose and low-dose SSD-treated (SSDH and SSDL). SSD was orally administrated of 1.50 mg/kg and 0.75 mg/kg/days for three weeks in the SSDH and SSDL groups, respectively. Fluoxetine was administrated at a dose of 2.0 mg/kg/days. SSD's antidepressant effects were assessed using the open field test, forced swim test, and sucrose preference test. Glutamate levels were quantified by ELISA. Western blot and immunochemical analyses were conducted to quantify proteins in the Homer protein homolog 1 (Homer1)-metabotropic glutamate receptor 5 (mGluR5) and mammalian target of rapamycin (mTOR) pathways in the hippocampal CA1 region. To measure related gene expression, RT-qPCR was employed. Results: CUMS-exposed rats treated with SSD exhibited increases in food intake, body weight, and improvements in the time spent in the central are and total distance traveled in the OFT, and less pronounced pleasure-deprivation behaviors. SSD also decreased glutamate levels in CA1. In CA1 region of CUMS-exposed rats, SSD treatment increased mGluR5 expression while decreasing Homer1 expression. SSD also increased expressions of postsynaptic density protein 95 (PSD95) and synapsin I (SYP), and the ratios of p-mTOR/mTOR, p-p70S6k/p70S6k, and p-4E-BP1/4E-BP1 in the CA1 region in CUMS-exposed rats. Conclusions: SSD treatment reduces glutamate levels in the CA1 region and promotes the expression of the synaptic proteins PSD-95 and SYP via the regulation of the Homer1-mGluR5 and downstream mTOR signaling pathways. These findings suggest that SSD could act as a natural neuroprotective agent in the prevention of depression. Highlights: Saikosaponin D, a pharmacologically active substance isolated from the Radix Bupleuri has been shown to have antidepressant effects. Its effects are mediated through the Homer1-mGluR5 and mTOR pathways. The findings support saikosaponin D's actions as a neuroprotective agent. [ABSTRACT FROM AUTHOR]

Diabetic retinopathy (DR) is the most common ocular fundus disease in diabetic patients. Chronic hyperglycemia not only promotes the development of diabetes and its complications, but also aggravates the occurrence of senescence. Previous studies have shown that DR is associated with senescence, but the specific mechanism has not been fully elucidated. Here, we first detected the differentially expressed genes (DEGs) and cellular senescence level of db/db mouse retinas by bulk RNA sequencing. Then, we used single-cell sequencing (scRNA-seq) to identify the main cell types in the retina and analyzed the DEGs in each cluster. We demonstrated that p53 expression was significantly increased in retinal endothelial cell cluster of db/db mice. Inhibition of p53 can reduce the expression of SA-β-Gal and the senescence-associated secretory phenotype (SASP) in HRMECs. Finally, we found that p53 can promote FoxO3a ubiquitination and degradation by increasing the expression of the ubiquitin-conjugating enzyme UBE2L6. Overall, our results demonstrate that p53 can accelerate the senescence process of endothelial cells and aggravate the development of DR. These data reveal new targets and insights that may be used to treat DR. • The expression of p53 was upregulated in the retina of db/db mice. • Retinal aging in db/db mice was significantly earlier than that in normal C57BL/6J mice. • Inhibiting p53 alleviates endothelial cell senescence. • p53 promotes the ubiquitination and degradation of FoxO3a through UBE2L6 and aggravates endothelial cell senescence. [ABSTRACT FROM AUTHOR]

Summary: T cells play crucial roles in the antitumour immune response. However, their dysfunction leads to inefficient tumour eradication. New members of the B7 family have moved to the fore of cancer research because of their involvement in T cell-mediated immune escape and tumorigenesis. Recently, bispecific antibodies (Bi-Abs) have become attractive because of their ability to activate T cells to target tumours. In this study, we examined the expression of new B7 family members B7-H4, B7-H5, B7-H6, and B7-H7 in human haematological tumour cells. Furthermore, we explored whether B7-H6 is an efficient target for T cell-induced cytotoxicity in haematologic malignant cells. We determined the capability of T cells armed with the bispecific antibody anti-CD3 × anti-B7-H6 (B7-H6Bi-Ab) to target haematological tumours in K562, Thp-1, Daudi, Jurkat, and U266 cells. Compared with their T cell counterparts, B7-H6Bi-Ab-armed T cells demonstrated significant cytotoxicity induction in B7-H6+ haematological tumour cells, according to quantitative luciferase and lactate dehydrogenase assays, and their activity was accompanied by increased levels of the secreted killing mediators granzyme B and perforin. Moreover, B7-H6Bi-Ab-armed T cells produced more T cell-derived cytokines: TNF-α, IFN-γ, and IL-2. In addition, compared to the control T cells, a higher level of the activation marker CD69 was detected on the B7-H6Bi-Ab-armed T cells. Taken together, these data suggest that the antitumour effect of B7-H6Bi-Ab-armed T cells may be a promising immunotherapy for use in future haematologic treatments. [ABSTRACT FROM AUTHOR]

This study identified 57 basic leucine zipper (bZIP) genes from the pineapple genome, and the analysis of these bZIP genes was focused on the evolution and divergence after multiple duplication events in relation to the pineapple genome fusion. According to bioinformatics analysis of a phylogenetic tree, the bZIP gene family was divided into 11 subgroups in pineapple, Arabidopsis, and rice; gene structure and conserved motif analyses showed that bZIP genes within the same subgroup shared similar intron-exon organizations and motif composition. Further synteny analysis showed 17 segmental duplication events with 27 bZIP genes. The study also analyzed the pineapple gene expression of bZIP genes in different tissues, organs, and developmental stages, as well as in abiotic stress responses. The RNA-sequencing data showed that AcobZIP57 was upregulated in all tissues, including vegetative and reproductive tissues. AcobZIP28 and AcobZIP43 together with the other 25 bZIP genes did not show high expression levels in any tissue. Six bZIP genes were exposed to abiotic stress, and the relative expression levels were detected by quantitative real-time PCR. A significant response was observed for AcobZIP24 against all kinds of abiotic stresses at 24 and 48 h in pineapple root tissues. Our study provides a perspective for the evolutionary history and general biological involvement of the bZIP gene family of pineapple, which laid the foundation for future functional characterization of the bZIP genes in pineapple. [ABSTRACT FROM AUTHOR]

Background: Oral squamous cell carcinoma (OSCC) is a solid tumor, which originates from squamous epithelium, with about 400,000 new-cases/year worldwidely. Presently, chemoradiotherapy is the most important adjuvant treatment for OSCC, mostly in advanced tumors. However, clinical resistance to chemotherapy still leads to poor prognosis of OSCC patients. Via high-throughput analysis of gene expression database of OSCC, we investigated the molecular mechanisms underlying cisplatin resistance in OSCC, analyzing the differentially expressed genes (DEGs) and their regulatory relationship, to clarify the molecular basis of OSCC chemotherapy resistance and provide a theoretical foundation for the treatment of patients with OSCC and individualized therapeutic targets accurately. Methods: Datasets related to "OSCC" and "cisplatin resistance" (GSE111585 and GSE115119) were downloaded from the GEO database and analyzed by GEO2R. Venn diagram was used to obtain drug-resistance-related DEGs. Functional enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis were performed on DEGs using The Database for Annotation, Visualization and Integrated Discovery (DAVID) software. Protein–protein interaction (PPI) network was constructed by STRING (search tool for recurring instances of neighbouring genes) database. Potential target genes of miRNA were predicted via miRDB, and cBioportal was used to analyze the function and survival of the potential functional genes. Results: Forty-eight upregulated DEGs and 49 downregulated DEGs were obtained from the datasets, with cutoff as p < 0.01 and |log FC| > 1. The DEGs in OSCC mainly enriched in cell proliferation regulation, and chemokine activity. In PPI network with hub score > 300, the hub genes were identified as NOTCH1 , JUN , CTNNB1 , CEBPA , and ETS1. Among miRNA–mRNA targeting regulatory network, hsa-mir-200c-3p, hsa-mir-200b-3p, hsa-mir-429, and hsa-mir-139-5p were found to simultaneously regulate multiple hub genes. Survival analysis showed that patients with high CTNNB 1 or low CEBPA expression had poor outcome. Conclusions: In the OSCC cisplatin-resistant cell lines, NOTCH1 , JUN , CTNNB1 , CEBPA , and ETS1 were found as the hub genes involved in regulating the cisplatin resistance of OSCC. Members of the miR-200 family may reverse drug resistance of OSCC cells by regulating the hub genes, which can act as potential targets for the treatment of OSCC patients with cisplatin resistance. [ABSTRACT FROM AUTHOR]

Summary: Prostate-specific antigen (PSA) has been widely used as the unique serum biomarker for the diagnosis of prostate cancer (PCa). When PSA is moderately increased (e.g., 4–10 ng/ml), it is difficult to differentiate benign prostatic hyperplasia (BPH) from cancer. The diagnostic test (i.e., prostate biopsy) is invasive, adding pain and economic burden to the patient. Urine samples are more convenient, non-invasive and readily available than blood. We sought to determine whether ferritin might be the potential urinary biomarker in prostate cancer diagnosis. Using two-dimensional electrophoresis (2DE) followed by mass spectrometry (MS), differentially expressed urinary proteins among patients with PCa, BPH and normal controls were obtained. The ferritin heavy chain (FTH) gene, ferritin light chain (FTL) gene and protein expression of BPH-1 cells and PC-3 cells were analyzed by real-time quantitative PCR and Western blotting, respectively. Stable FTH or FTL silenced cell lines were generated by small hairpin(sh) RNA lentiviral transfection. The function of the cell lines was evaluated by the colony formation assay, transwell assay, and flow cytometry. Compared with BPH and normal controls, 15 overexpressed proteins, including FTH and FTL, were identified in the urine of the PCa group. FTH and FTL were also highly expressed in PC-3 cell lines compared with BPH-1 cells. FTH-silenced cells showed reduced cell proliferation, migration and increased cell apoptosis. FTL-silenced cells showed increased proliferation and migration abilities. There are differences in urinary proteins among patients with PCa, BPH and normal controls. FTH and FTL play different roles in PCa cells and are potential biomarkers for PCa. [ABSTRACT FROM AUTHOR]

Forkhead box‐O1 (FoxO1) is a key nutrient‐ and growth factor‐dependent regulator of metabolism, but its functional role in human primary keratinocytes (HPKs) is less known. To investigate the role of FoxO1 in HPKs and effect of insulin‐like growth factor 1 (IGF‐1) and isotretinoin on FoxO1 expression, HPKs were treated with 1.2 mmol/L calcium chloride, 1‐20 ng/mL IGF‐1 and 0.1‐10 μmol/L isotretinoin. Recombinant adenovirus expressing FoxO1 or FKHR shRNA lentivirus transfection was introduced to upregulate or silence FoxO1 expression. Epidermal FoxO1 immunostaining was lower in acne lesion than in normal skin. FoxO1 overexpression induced involucrin expression, G2/M arrest and apoptosis but suppressed proliferation, while FoxO1 silencing decreased involucrin expression but increased proliferation, S phase and viable cells in HPKs. IGF‐1 downregulated FoxO1 and involucrin but upregulated p‐Akt expression in HPKs, which was blocked by pretreatment with LY294002. Isotretinoin enhanced FoxO1, p53 and p21 but inhibited p‐FoxO1 and involucrin expression in HPKs. These results demonstrate that FoxO1 promotes differentiation and apoptosis in HPKs. IGF‐1 may reduce keratinocyte differentiation through PI3K/Akt/FoxO1 pathway, while isotretinoin can reinforce FoxO1 expression. FoxO1 may be involved in acne pathogenesis and could serve as a potential therapeutic target. [ABSTRACT FROM AUTHOR]

Aims: Fusarium oxysporum is a causal disease that threatens watermelon production, but little information on the molecular mechanisms involved in host defense is available. To understand the defense response, a proteome-level changes that occur in watermelon roots during F. oxysporum infection were investigated.Methods: We utilized two-dimensional gel electrophoresis (2-DE) to compare changes in the root proteome profiles and validated their expression using real-time PCR.Results: A total of 690 spots were detected, and 32 proteins had significant changes in abundance and were further identified by mass spectrometry. These proteins were mainly involved in metabolism, stress and defense and amino acid biosynthesis. RT-PCR analysis revealed that transcripts corresponding to the nine randomly selected proteins could be significantly induced, their expression patterns were consistent with the proteomic results except for Apx and Tdh. The involvement of these proteins in regulating watermelon response against F. oxysporum is discussed.Conclusions: The reprogrammed proteins were involved in several biological processes, which indicates that watermelon can directly alter the abundance of these proteins to establish a defense response. This work helps us understand the basic processes during the watermelon-F. oxysporum interaction and may contribute to improve resistance breeding toward this pathogen. [ABSTRACT FROM AUTHOR]

Inducible NO synthase (iNOS) expression and peroxynitrite formation are significantly increased in diabetic vascular tissues. Transcription factor KLF5 activates iNOS gene transcription and is involved in vascular inflammatory injury and remodeling. However, mutual regulation between KLF5, iNOS and peroxynitrite in diabetic vascular inflammation, as well as the underlying mechanisms, remain largely unknown. In this study, we found a marked increase in KLF5 and iNOS expression in vascular smooth muscle cells (VSMC) of diabetic patients. High glucose-induced expression of KLF5 and iNOS was also observed in cultured mouse VSMCs. Further investigation showed that high glucose induced KLF5 nitration by iNOS-mediated peroxynitrite generation, and nitrated KLF5 increased its interaction with NF-κB p50 and thus cooperatively activated the expression of inflammatory cytokines TNF-α and IL-1β. Furthermore, we showed that the VSMC-specific knockout of KLF5 dramatically reduced inflammatory cytokine expression in the vascular tissues of diabetic mice. Moreover, 17β-estradiol (E2) inhibited high glucose-mediated effects in VSMCs, and in the response to E2, estrogen receptor (ER) α competed with KLF5 for binding to NF-κB p50, which in turn leads to the suppression of inflammatory gene expression in VSMCs. Together, the present findings were the first to show that KLF5 expression and nitration by iNOS-mediated peroxynitrite are necessary for the induction of TNF-α and IL-1β expression in VSMCs of diabetic vascular tissues. [ABSTRACT FROM AUTHOR]

The comprehensive analysis of single or multiple microarray datasets is currently available in Gene Expression Omnibus (GEO) databases, with several studies having identified genes strongly associated with the development of lung adenocarcinoma (LUAD). However, the mechanisms of LUAD development remain largely unknown and has not yet been systematically studied; thus, further studies are required in this field. In the present study, weighted gene co-expression network analysis (WGCNA) was used for the evaluation of key genes with potential high risk of LUAD, and to provide more reliable evidence concerning its pathogenesis. The GSE140797 dataset from the high-throughput GEO database was downloaded and was first analyzed using the Limma package in the R language in order to determine the differentially expressed genes. The dataset was then analyzed using the WGCNA package to analyze the co-expressed genes, and the modular genes with the highest correlation with the clinical phenotype were identified. Subsequently, the pathogenic genes shared in common between the result of the two analyses were imported into the STRING database for protein-protein interaction network analysis. The hub genes were screened out using Cytoscape, and then The Cancer Genome Atlas analysis, receiver operating characteristic analysis and survival analysis were subsequently performed. Finally, the key genes were evaluated using reverse transcription-quantitative PCR and western blot analysis. Bioinformatics analysis of the GSE140797 dataset revealed eight key genes: AURKA, BUB1, CCNB1, CDK1, MELK, NUSAP1, TOP2A and PBK. Finally, the AURKA, TOP2A and MELK genes were evaluated in samples from patients with lung cancer using WGCNA and RT-qPCR, western blot analysis experiments, providing basis for further research on the mechanisms of LUAD development and targeted therapy. [ABSTRACT FROM AUTHOR]

Toll-like receptor (TLR)-mediated signaling are critical for host defense against pathogen invasion. However, excessive responses would cause harmful damages to the host. Here we show that deficiency of the E3 ubiquitin ligase TRIM32 increases poly(I:C)- and LPS-induced transcription of downstream genes such as type I interferons (IFNs) and proinflammatory cytokines in both primary mouse immune cells and in mice. Trim32-/- mice produced higher levels of serum inflammatory cytokines and were more sensitive to loss of body weight and inflammatory death upon Salmonella typhimurium infection. TRIM32 interacts with and mediates the degradation of TRIF, a critical adaptor protein for TLR3/4, in an E3 activity-independent manner. TRIM32-mediated as well as poly(I:C)- and LPS-induced degradation of TRIF is inhibited by deficiency of TAX1BP1, a receptor for selective autophagy. Furthermore, TRIM32 links TRIF and TAX1BP1 through distinct domains. These findings suggest that TRIM32 negatively regulates TLR3/4-mediated immune responses by targeting TRIF to TAX1BP1-mediated selective autophagic degradation. [ABSTRACT FROM AUTHOR]

Rootstock grafting may improve the resistance of watermelon plants to low temperatures. However, information regarding the molecular responses of rootstock grafted plants to chilling stress is limited. To elucidate the molecular mechanisms of chilling tolerance in grafted plants, the transcriptomic responses of grafted watermelon under chilling stress were analyzed using RNA-seq analysis. Sequencing data were used for digital gene expression (DGE) analysis to characterize the transcriptomic responses in grafted watermelon seedlings. A total of 702 differentially-expressed genes (DEGs) were found in rootstock grafted (RG) watermelon relative to self-grafted (SG) watermelon; among these genes, 522 genes were up-regulated and 180 were down-regulated. Additionally, 164 and 953 genes were found to specifically expressed in RG and SG seedlings under chilling stress, respectively. Functional annotations revealed that up-regulated DEGs are involved in protein processing, plant-pathogen interaction and the spliceosome, whereas down-regulated DEGs are associated with photosynthesis. Moreover, 13 DEGs were randomly selected for quantitative real time PCR (qRT-PCR) analysis. The expression profiles of these 13 DEGs were consistent with those detected by the DGE analysis, supporting the reliability of the DGE data. This work provides additional insight into the molecular basis of grafted watermelon responses to chilling stress. [ABSTRACT FROM AUTHOR]

Chemotherapy is an important strategy for the treatment of bladder cancer. However, the main problem limiting the success of chemotherapy is the development of multidrug resistance (MDR). To improve the management of bladder cancer, it is an urgent matter to search for strategies to reverse MDR. We chose three kinds of herbal medicines including ginsenoside Rh2, (-)-Epigallocatechin gallate (EGCG) and Tetramethylpyrazine (TMP) to detect their effects on bladder cancer. Reversal effects of these three herbal medicines for drug resistance in adriamycin (ADM)-resistant Pumc-91 cells (Pumc-91/ADM) were assessed by Cell Counting Kit-8 (CCK-8) cell proliferation assay system. The mechanisms of reversal effect for TMP were explored in Pumc-91/ADM and T24/DDP cells. After Pumc-91/ADM and T24/DDP cells were treated with TMP, cell cycle distribution analysis was performed by flow cytometry. The expression of MRP1, GST, BCL-2, LRP and TOPO-II was evaluated using quantitative real-time , immunefluorescence assay and western blot. It was observed that TMP was capable of enhancing the cytotoxicity of anticancer agents on Pumc-91/ADM cells in response to ADM, however Rh2 and EGCG were unable to. The reversal effect of TMP was also demonstrated in T24/DDP cells. Moreover, the treatment with TMP in Pumc-91/ADM and T24/DDP cells led to an increased of G1 phase accompanied with a concomitant decrease of cell numbers in S phase. Compared to the control group, an obvious decrease of MRP1, GST, BCL-2 and an increase of TOPO-II were shown in TMP groups with a dose-dependency in mRNA and protein levels. However, there was no difference on LRP expression between TMP groups and the control group. TMP could effectively reverse MDR of Pumc-91/ADM and T24/DDP cells and its mechanisms might be correlated with the alteration of MRP1, GST, BCL-2 and TOPO-II. TMP might be a potential candidate for reversing drug resistance in bladder cancer chemotherapy. [ABSTRACT FROM AUTHOR]

This study aimed to identify the highest efficiency promoter sequence of β-lactoglobulin (BLG) gene and construct a mammary-specific expression vector for expressing high level of transgene in goat mammary epithelial cells. We amplified 13 BLG proximal promoter fragments, 2 introns, and a 3′ untranslated region from goat genome. The dual-luciferase assay showed that the promoter activities of these BLG proximal promoter fragments were significantly higher in HC11 than those in 293T or GEF cells, and that BLG promoter region BLG11 (431 bp, −391/+40) was an optimal sequence to drive transgenic expression. Western blot analysis confirmed that the promoter sequence BLG11 had a higher promoter activity in goat mammary epithelial cells than other promoter sequences. Moreover, the expression levels of GFP were significantly increased by combining promoter region BLG11 and native intron 1 in transfected goat mammary epithelial cells, and increased further with introns 1 and 2. This study demonstrated that a short BLG promoter region BLG11 combined with introns 1 and 2 could promote transgenic expression in goat mammary epithelial cells. These mammary regulated elements may be useful for production of mammary gland bioreactor. [ABSTRACT FROM AUTHOR]

The essence of the reprogramming activity of somatic cell nuclear transfer (SCNT) embryos is to produce normal fertilized embryos. However, reprogramming of somatic cells is not as efficient as the reprogramming of sperm. In this report, we describe the effect of an inducible, specific miR-34 microRNA expression in donor cells that enables a similar level of sperm:transgene expression on the early development of SCNT embryos. Our results showed that donor cells with doxycycline (dox)-induced miR-34c expression for the preparation of SCNT embryos resulted in altered developmental rates, histone modification (H3K9ac and H3K4me3), and extent of apoptosis. The cleavage rate and blastocyst formation of the induced nuclear transfer (NT) group were significantly increased. The immunofluorescence signal of H3K9ac in embryos in the induced NT group significantly increased in two-cell- and eight-cell-stage embryos; that of H3K4me3 increased significantly in eight-cell-stage embryos. Although significant differences in staining signals of apoptosis were not detected between groups, lower apoptosis levels were observed in the induced NT group. In conclusion, miR-34c expression induced by dox treatment enhances the developmental potential of SCNT embryos, modifies the epigenetic status, and changes blastocyst quality. [ABSTRACT FROM AUTHOR]

Abstract: Marsupenaeus japonicus is a shrimp species of great value in the Chinese aquaculture industry. Given the susceptibility to viral diseases, research efforts have focused on the molecular characteristics of the shrimp's immune mechanisms. Ferritin is well known for its iron storage function, but studies have also addressed its immune function in response to pathogens. In this study, an M. japonicus ferritin cDNA was identified by homology cloning and rapid amplification of cDNA ends-PCR. The full-length cDNA is 1244bp long and contains an open reading frame (513bp) that encodes a highly conserved protein of 170 amino acids. Quantitative real-time PCR detection of ferritin revealed high expression in eight tested tissues, with the highest levels in hemocytes—consistent with the iron storage capacity of ferritin. We infected M. japonicus with white spot syndrome virus and validated the model by viral copy analysis and histopathology, which demonstrated an increase in viral copies along with acute degeneration of tissues. Transcripts of ferritin increased by 3.1-fold, 2.1-fold, and 1.5-fold in the hepatopancreas, gill, and midgut at 24h post-injection, suggesting that ferritin played an important role in the immune response of M. japonicus. [Copyright &y& Elsevier]

Purpose: Resistance to chemotherapy drugs remains a difficult problem in bladder cancer treatment. Protein expression is an important factor underlying multidrug resistance (MDR) in bladder cancer. The aim of the study was to explore differentially expressed proteins responsible for MDR between an adriamycin-resistant human bladder cancer cell line (pumc-91/ADM) and its parental cell line (pumc-91). Methods: Two-dimensional gel electrophoresis (2-DE) combining image analysis was used to screen the differentially expressed protein spots between the pumc-91/ADM and pumc-91 cell lines. Then, the protein spots were identified using MALDI-TOF/TOF mass spectrometry. Among the identified proteins, annexin A2 (ANXA2) and nucleophosmin (NPM1) were then further verified using RT-PCR and Western blot analysis. Results: A total of 30 proteins, including 19 up-regulated and 11 down-regulated proteins, were successfully identified in pumc-91/ADM. According to their different functions, these 30 proteins were classified into 12 categories. Annexin A2 (ANXA2) and nucleophosmin (NPM1) were up-regulated in pumc-91/ADM compared with pumc-91. Conclusion: The proteins identified may have an important clinical significance in MDR, and ANXA2 and NPM1 may take part in mechanism of MDR in bladder cancer. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effect of dietary L-theanine supplementation on skeletal muscle fiber type transition in mice. Our data indicated that dietary 0.15% L-theanine supplementation significantly increased the mRNA expression levels of muscle fiber type related genes (MyHC I, MyHC IIa, PGC-1α, Sirt1, Tnnt1, Tnnc1, Tnni1, MEF2C) and the protein expression levels of MyHC IIa, myoglobin, PGC-1α, Sirt1 and Troponin I-SS, but significantly decreased the mRNA and protein expression levels of MyHC IIb. Dietary 0.15% L-theanine supplementation significantly increased the activities of SDH and MDH and decreased the activity of LDH. Furthermore, immunofluorescence demonstrated that dietary 0.15% L-theanine supplementation significantly increased the percentage of type I fibers, and significantly decreased the percentage of type II fibers. In addition, we found that dietary 0.15% L-theanine supplementation increased the fatigue-resistant, antioxidant capacity, mitochondrial biogenesis, and function in skeletal muscle of mice. Furthermore, dietary 0.15% L-theanine supplementation significantly increased the mRNA levels of prox1, CaN and NFATc1, the protein levels of prox1, CNA and NFATc1 and the activity of CaN in GAS muscle when compared with the control group. These results indicated that dietary L-theanine supplementation promoted skeletal muscle fiber transition from type II-type I, which might be via activation of CaN and/or NFATc1 signaling pathway. [ABSTRACT FROM AUTHOR]

Inflammatory bowel disease (IBD) is one of the most common chronic inflammatory illnesses of the gastrointestinal tract due to the imbalance of immune homeostasis of T helper cells and/or regulatory T cells (Tregs). The Traditional Chinese medicine herb has been clinically proven for use in the treatment of IBD but its possible mechanism remains unknown. The study aims to assess the effect of Chinese medicinal herb decoction QRZSLXF (Qing Re Zao Shi Liang Xue receipt) for the treatment of TNBS-induced experimental colitis in mice and explore its relevant mechanism involved in Th17 and Tregs. Mice colitis was induced by 50% 2,4,6-Trinitrobenzenesulfonic Acid (TNBS) ethanol solution weekly manner. These established model mice were divided into model control (0.8% NaCl treatment), FICZ, naphthoflavone (NaFTV), dexamethasone (DXM), and QRZSLXF (QrLx) groups. The colonoscopy, H&E staining, and immune staining were used to analyze the disease severity, inflammatory condition and Th17 or Treg related factors expression. High-performance liquid chromatography-mass spectrometry (HPLC/MS) was used to assess the content of FICZ in the colon tissues. Western blot and ELISA were used to examine the expression of Th17 or Treg related factors protein levels. Flow cytometry analysis was performed to assess the number and ratio of Th17/Tregs in splenocytes, and mesenteric lymph node lymphocytes (MLNCs), and lamina propria mononuclear cells (LPMCs). NaFTV, DXM and QrLx groups intestinal inflammation scores were significantly lower than that in colitis model control and FICZ groups, while the IL-6, STAT3, and RORγt expression levels were significantly lower than those in the model control and FICZ groups. Mass spectrometry results showed FICZ that in both DXM and QrLx groups was lower than control model and FICZ groups. Flow cytometry results showed that DXM, NaFTV and QrLx could significantly reduce Th17 proportion and increase Treg proportion in splenocytes, MLNCs, and LPMCs. NaFTV and QrLx treatment could decrease symptoms and inflammatory colitis, by decreasing of FICZ concentration and AhR signaling in colon, resulting in reducing the expression of IL-6, STAT3, and RORγt, whereas increasing the expression of FOXP3, consequently reducing the proportion of Th17 cells and increasing the proportion of Treg cells, respectively. Image 1 [ABSTRACT FROM AUTHOR]

• Adult zebrafish are more sensitive to F-53B than embryos, with a lower 96 h LC50. • F-53B exposure impairs the gill structure of zebrafish. • F-53B exposure leads to an increase in erythrocyte numbers in zebrafish. • F-53B decreases egg production and increases the GSI in zebrafish. • F-53B disrupts steroidogenic gene expression and sex hormone production in zebrafish. 6:2 Chlorinated polyfluorinated ether sulfonate, commonly known as F-53B, is widely used as a mist suppressant in various industries and is frequently detected in the environment. Despite its prevalent presence, the adverse effects of F-53B are not well understood and require future investigation. This study utilized zebrafish embryos and adults to examine the toxic effects of F-53B. Our findings revealed that F-53B impaired gill structure and increased erythrocyte numbers in adult zebrafish. Notably, F-53B demonstrated a higher sensitivity for inducing mortality (LC 50 at 96 h) in adult zebrafish compared to embryos. Additionally, F-53B disrupted the expression of critical steroidogenic genes and hindered sex hormone production, which negatively affecting egg production. In conclusion, this study underscores the detrimental impact of F-53B on gill structure and reproductive toxicity in zebrafish, providing valuable insights into its overall toxicity. [ABSTRACT FROM AUTHOR]

Major histocompatibility complex II (MHCII) is a critical immune molecule that plays an essential role in combating external pathogens in fish. To investigate MHCII in the economically significant fish species, silver pomfret, we firstly identified three PaMHCII s, including one PaMHCIIa and two PaMHCIIb s, all closely related to Trachinotus ovatus MHCII. Ig superfamily, C1-set and IGc1 domain were identified in PaMHCII s, with MHCII alpha and MHCII beta domians found in PaMHCIIa and PaMHCIIb s, respectively. To further elucidate their immune function, we analyzed three transcriptomes from fish infected with a primary pathogenic bacterium, discovering that MHCII was highly enriched in Antigen processing and presentation (map04612). To explore the regulatory relationships of these differentially expressed genes (DEGs) in vitro , we established a Pampus argenteus kidney cell line (PaK) and construct a main pathogenic bacteria- Photobacterium damselae subsp. Damselae (PDD) infection PaK model according to the change of cell viability. In this model, the TUNEL signal did not exhibit significant changes at 6 h but increased gradually; compared to 0 h, tested genes were up-regulated at 6 h; All genes, except PaMHCII s, GILT and CTSB, were up-regulated at 12 h. At 18 h, TCR was up-regulated while Ii and CTSL were significantly down-regulated. By 24 h, TCR , Ii and CTSL were down-regulated. These results suggest that the MHCII pathway is activated during the early stages of infection, but PDD may suppress it later through immune evasion mechanisms. This data enhances our understanding of the origin, evolution, and function of MHCII and provides guidance for disease control in silver pomfret aquaculture. • We identified one MHCII a and two MHCII b s in silver pomfret. • We constructed Pampus argenteus kidney cell line (PaK). • MHCII were enriched in MHCII pathway in transcriptomes under exogenous stresses. • Acquired immunity regulated by MHCII was activated in early PDD infection in vitro. • PDD depressed the whole antigen processing and presentation through immune escape. [ABSTRACT FROM AUTHOR]

Members of the Signal Transducer and Activator of Transcription (STAT) family function pivotally as transcriptional activators integral to the modulation of inflammatory responses. The aquaculture of silver pomfret is frequently compromised by the imposition of exogenous stressors, which include thermal fluctuations, notably low-temperatures, diminished oxygen levels, and the onslaught of bacterial pathogens. Notwithstanding the critical impact of these stressors, the scientific literature presents a notable gap in our understanding of the STAT pathway's role in the silver pomfret's adaptive response mechanisms. To address this lacuna, we identified stat genes in the silver pomfret—denominated as Pastat1 , Pastat2 , Pastat3 , Pastat4 , and Pastat5 —through a thorough and systematic bioinformatics analysis. Further scrutiny of the gene configurations and constituent motifs has elucidated that STAT proteins possess analogous structural frameworks and exhibit significant evolutionary preservation. Subsequently, the expression patterns of five stat genes were verified by RT-qPCR in twelve different tissues and four growth periods in healthy fish, showing that the expression of Pastat genes was temporally and spatially specific, with most of the stat genes expressed at higher levels in the spleen, following muscle, gill, and liver. Transcriptomic analysis of exposure to exogenous stressors, specifically formaldehyde and low-temperature conditions, elucidated that Pastat1 and Pastat2 genes exhibited a heightened sensitivity to these environmental challenges. RT-qPCR assays demonstrated a marked alteration in the expression profiles of jak1 and Pastat gene suites in PaS upon prolonged bacterial infection subsequent to these exogenous insults. Moreover, the gene expression of the downstream effectors involved in innate immunity and apoptosis displayed marked deviations. This study additionally elucidated the Pastat gene family's role in modulating the innate immune response and apoptotic regulation within the silver pomfret during exogenous stressors and subsequent pathogenic incursions. • There have five stat family genes in silver pomfret. • The changes of stat genes expression in response to exogenous stresses were complex. • Significant changes in stat genes expression in bacterial infection after exogenous stresses. • Stat genes are involved in innate immunity and apoptosis-related pathways. [ABSTRACT FROM AUTHOR]

Jellyfish venom can cause the mass mortality fish and induces apoptosis of many types of cells; therefore, we speculated that the venom might be able to induces apoptosis in fish. To further understand the mechanism, we cultured Larimichthys crocea with Rhopilema esculentum and performed transcriptome analysis of liver, heart, and kidney of the fish. According to KEGG analysis, we selected certain apoptosis-related pathways, such as the P53 signaling pathway and apoptosis, and analyzed the apoptosis-relatedrelated DEGs in these pathways in the three tissues using quantitative real-time reverse transcription PCR. PIG3 , CTSL , CytC , AP1 and Gadd45a were all upregulated in three tissues; whereas Mdm4 was downregulated in heart tissues. Then, we treated HEK 293 T cells with jellyfish venom and knocked down the expression of PIG3 and CTSL using small interfering RNAs (siRNAs). In cells treated with jellyfish venom, cell viability increased significantly after RNAi; P53 and CASP9 were upregulated, but the expression of CASP9 was much higher in the Control-RNAi group (without siRNA). PIG3 and BAX were downregulated in the PIG3 -RNA+ (with PIG3 siRNA); CTSL and CYTC were upregulated, but BCL2 was downregulated in the Control-RNAi group. The protein levels of PIG3, CTSL, and CASP9 detected by western blotting showed similar trends to their mRNA expression levels. Thus, the results indicated that jellyfish venom could activate PIGs through the P53-PIG3 pathway to damage lysosomes, which released CTSL to affect mitochondria, leading to the apoptosis in fish. The data increases our understanding of the mechanism of apoptosis induced by jellyfish venom. [Display omitted] • Jellyfish venom can cause mortality of fish and apoptosis of many model cells. • Apoptosis relating genes were highly expressed in fish treated by jellyfish venom. • RNAi depressed the expression of Cyt C and Casp9 in cell treated by jellyfish venom. • Jellyfish venom could induced apoptosis of fish by PIG3 of P53-PIG3-CTSL pathway. • The data increases understanding of mechanism for cytotoxicity of jellyfish venom. [ABSTRACT FROM AUTHOR]

• Characterizing distinct metabolic and transcriptional profiles associated with raw materials of Geqingpi, Sihuaqingqi and Citri Reticulatae Pericarpium. • The relative content of flavonoids in the pericarp of Citrus reticulata 'Chachi' is the most abundant, with polymethoxyflavonoids (PMFs) being the main component of flavonoids. • A set of seven biomarkers were identified that demonstrated the ability to clearly distinguish between the three stages, and nobiletin possessed the most significant contribution. • A total of 93 O -methyltransferase genes were identified in Citrus reticulata 'Chachi', and 28 of them were potentially responsible for catalyzing the biosynthesis of PMFs (i.e., sinensetin, nobiletin and 3,5,6,7,8,3',4'- heptemthoxyflavone). The dried pericarp of Citrus reticulata 'Chachi' (CRC) is commonly utilized in Traditional Chinese Medicine (TCM) and food consumption. Geqingpi (GQP), Sihuaqingpi (SHQP) and Guangchenpi (CRP) are TCM materials derived from different developmental stages of CRC pericarp, each possessing distinct medicinal properties. However, there is currently limited study on the active substances in these three stages. Metabolome data suggested that the discrepancies in flavonoid content, particularly polymethoxyflavonoids (PMFs), predominantly contribute to the variations across the three periods. Additionally, through multivariate statistical analysis, 7 potential biomarkers were identified and nobiletin (NOB) possessed the most significant contribution. O -methyltransferases (OMTs) play a crucial role to catalyze the formation of -OCH 3 in PMFs. The analysis of transcriptome and genome data revealed the presence of 93 CrOMTs in CRC and 43 were expressed on pericarp. Further investigation identified that 28 of these genes were potentially responsible for catalyzing the biosynthesis of important PMFs such as NOB, sinensetin (SIN), and 3,3′,4′,5,6,7,8-heptamethoxyflavone (HEP). Our results may provide a new perspective for the study of GQP, SHQP and CRP, as well as the regulatory mechanism of PMFs in CRC. [ABSTRACT FROM AUTHOR]

Photobacterium damselae subsp. damselae (PDD) is a dangerous bacterial disease that affects silver pomfret (Pampus argenteus) and its main virulence factors are pore-forming toxins. In this study, a highly pathogenic strain was obtained from naturally infected silver pomfret and was introduced into healthy fish for experimentation. Liver, gill, kidney and spleen samples were collected at 0, 24, 48, 72 and 96 h. Histopathological evaluations of these tissues, via HE staining, showed multi-organ bleeding, inflammatory infiltration and histopathocyte vacuolation as the main pathological changes. The levels of AKP, ACP, LZM, SOD, GSH and CAT in serum also displayed an increase following infection. Transcriptome sequencing of liver, kidney and spleen was performed using samples 96 h post-infection. The analysis of the resulting 770,878,120 clean reads collectively assembled into 89,310 unigenes. The WGCNA analysis demonstrated enrichment of phagosome and cytokine-cytokine receptor interaction in the liver, drug metabolism-cytochrome P450 in the kidney, and MAPK signaling pathway and other signaling pathways in the spleen. DEGs related to immune and immune-related pathways, such as Phagosome, Autophagy-animal and Glycerolipid metabolism, were significantly enriched. Selected key genes from these KEGG pathways were examined using RT-qPCR. The expression level of autophagy related genes (pi3k, bcl2) increased after infection, while the gene associated with MAPK and apoptosis (Caspase 9, mapk4) significantly increased after 72 h infection. The expression of phagosome related genes (atp6v1f, ctsl) increased initially, but later decreased. The genes related to innate immunity (ccl4, il15) were up-regulated, while those related to acquired immunity (mhc2a, mhc2b) showed a downward trend. Lysosome-related genes (gilta, giltb, cftr, c-type lectin, m6pra and m6prb) were up-regulated in liver and spleen and down-regulated in kidney and gill. The data obtained illustrated that PDD activated the immune defenses of the fish, increasing metabolic expenditure, but that the pore-forming toxins released by PDD could destroy the phagosome membrane and help the bacteria escape, which could disturb antigen presentation and inhibit the activation of the acquired immunity. This study reveals the potential immune escape mechanism of PDD in the host for the first time and has the potential to aid in the development of anti-PDD drugs. • PDD infection caused multi-organ bleeding, inflammatory infiltration in fish tissues. • Immune relating enzyme activities increased after PDD infection. • PDD activated the immune defenses and increased metabolic expenditure. • Pore-forming toxins of PDD might destroy phagosome and help PDD escape. • This immune escape might disturb antigen presentation and inhibit acquired immunity. [ABSTRACT FROM AUTHOR]

• Copper bioaccumulation in fish led to tissue damage and abnormal collagen metabolism. • PaM was constructed for the first time as a research tool for environmental pollution. • Copper disrupted mediation of timp -MMPs for ECM balance through AKTs/ERK/FGFs. Copper pollution might have a negative effect on collagen metabolism in fish. To test this hypothesis, we exposed an important economical fish, silver pomfret (Pampus argenteus), to three concentrations of Cu2+ for up to 21 days to simulate natural exposure to copper. With increasing copper exposure concentration and time, hematoxylin and eosin staining and picrosirius red staining revealed extensive vacuolization, cell necrosis, and tissue structure destruction, and a change of type and abnormal accumulation of collagen in the liver, intestine, and muscle tissues. To further study the mechanism of collagen metabolism disorder caused by copper exposure, we cloned and analyzed a key collagen metabolism regulation gene, timp , of silver pomfret. The full-length timp2b cDNA was 1035 bp with an open reading frame of 663 bp, encoding a protein of 220 amino acids. Copper treatment significantly increased the expression of akts, erks , and fgfs genes and decreased the mRNA and protein expression of Timp2b and MMPs. Finally, we constructed a silver pomfret muscle cell line (PaM) for the first time and used PaM Cu2+ exposure models (450 μM Cu2+ exposure for 9 h) to examine regulation function of the timp2b-mmps system. We knocked down or overexpressed timp2b in the model, and found that downregulation of mmps expression and upregulation of akt/erk/fgf were further aggravated in the timp2b − group (subjected to RNA interference), whereas some recovery was achieved in the timp2b + group (overexpression). These results indicated that long-term excessive copper exposure can lead to tissue damage and abnormal collagen metabolism in fish, which might be caused by the alteration of akt/erk/fgf expression, which disrupts the effects of the timp2b-mmps system on extracellular matrix balance. The present study assessed the impact of copper on the collagen of fish and clarified its regulatory mechanism, providing a basis for toxicity of copper pollution study. [ABSTRACT FROM AUTHOR]

Abstract: Purpose: The transforming growth factor β (TGF-β) signaling pathway is involved in the epileptogenesis. Sorting Nexin 25 (SNX25) has been recently proposed to modulate TGF-β signaling through endosomal sorting of TGF-β receptors for lysosomal degradation. The aim of the present study was to determine the expression pattern of SNX25 in brains of epilepsy patients and in animal model of epilepsy. Methods: We investigated the expression of SNX25 in the brain tissues of patients with temporal lobe epilepsy (TLE) and in the pilocarpine-induced rat model of epilepsy using western blotting, real-time quantitative RT-PCR, and double-label immunofluorescence. Results: The expression of SNX25 was significantly increased in TLE patients in comparison to controls (0.21±0.07 vs. 0.11±0.03, P<0.05). In the lithium-pilocarpine induced epileptic rats, significant elevation of SNX25 levels was detected in the chronic phase, while no SNX25 alteration occurred in the acute and latent phases. Moreover, SNX25 localized to astrocytes and neurons, in both human samples and animal models. Conclusion: Our results indicate that upregulation of SNX25 might be involved in the development of temporal lobe epilepsy. [Copyright &y& Elsevier]

Abstract: Infectious bursal disease virus (IBDV) can cause disease in chickens characterized by immunosuppression and high mortality. Currently, real-time RT-PCR has been used to quantitate virus-specific RNA and to better understand host response to infection. However, normalization of quantitative real-time RT-PCR is needed to a suitable internal control. We thus investigated the expression pattern of six chicken genes, including β-actin, 28S rRNA, 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), TATA box-binding protein (TBP) and β-2-microglobulin, in chicken embryo (CE) cell cultures following a 7-day IBDV infection. The CE cells were inoculated with various multiplicity of infection (MOI) of IBDV vaccine strain Bursine-2, the expression of genes was measured by quantitative real-time PCR-based on cDNA synthesized from either normalized (100ng) or non-normalized (10μl) total RNA. The results showed that β-actin, 28S rRNA, 18S rRNA and GAPDH were the most constantly expressed genes, while TBP and β-2-microglobulin were markedly induced during the infection course. Of these constant expressed genes, 28S rRNA and 18S rRNA are highly expressed; β-actin intermediately expressed and GAPDH had a lower expression level in CE cell cultures. Also, β-actin showed no significant variation in both normalized and non-normalized assays and virus dose-independent of inoculation, while other genes did. β-Actin was further successfully used as an internal control to quantitate Bursine-2 virus-specific RNA load in CE cell cultures. Thus, β-actin was suggested as a suitable internal control in studying gene expression as well as virus-specific RNA load in CE cell after IBDV infection. [Copyright &y& Elsevier]

• Exposure to 30 μg/L HgCl2 up-regulated 1,788 and down-regulated 469 genes. • Most DEGs were related to xenobiotic biodegradation, development, oxidative stress. • Two metabolic pathways were identified to be key for HgCl 2 metabolism. • 10 DEGs were identified as key HgCl 2 -responsive genes. • We firstly investigated transcriptional changes to HgCl 2 in zebrafish intestine. Mercury is a widely used heavy metal that causes pollution to aquatic environments and severely affects the health of fish. Little is known about how heavy metal pollutants affect fish, particularly for gene expression within important organs such as the intestine. Herein, whole transcriptome sequencing was performed on zebrafish (Danio rerio) intestine tissue after HgCl 2 (HGC, 30 μg/L) exposure. A total of 2,257 differentially expressed genes (DEGs) were identified, including 1,788 up- and 469 down-regulated genes. Functional enrichment analysis revealed that these DEGs were primarily related to xenobiotic biodegradation, biomacromolecule metabolism, development, oxidative defense, and immune response. Ten key HGC-responsive DEGs were screened to survey the dynamic changes of expression in response to HGC exposure at different time points, and were also used to validate RNA sequencing data using quantitative real-time PCR (qPCR). Results indicate that the expression of genes encoding UGT1AB, GSTT1B, GSTO1, GSTM2, UGT5G1, GSTT1A, GSTR, GSTM3, GSTA1, and GSTP2 were significantly upregulated in response to the HGC exposure, and potentially help to counteract the adverse effects of HGC. This study provides insight into fish molecular toxicological responses to heavy metals and method on environmental risk assessment. [ABSTRACT FROM AUTHOR]

p-Nitrophenol (PNP) is one type of environmental pollutant, which is difficult to degrade and soluble in water. To investigate the effects of PNP on economically important marine fish species, we subjected Larimichthys crocea juvenile to five different concentrations of PNP for 96 h, and the semi-lethal concentration (LC50) was 6.218 mg/L. Then we collected the liver, kidney, and gill tissues to determine enzyme activity and gene expression levels, and analyzed histological changes. In histological analysis, the gills showed curling of lamella, epithelial lifting and hyperplasia; the parenchymal structure of hepatocytes was significantly damaged, with severe vacuolation and loss of original structure. The renal cells were damaged too, with congestion and renal tubular necrosis. Catalase and superoxide dismutase both showed an up- and down-tendency with the rise of concentration in the three tissues, and GSH-px had similar trend in the kidney, which decreased at 8 mg/L in the liver but showed no significant differences in the gills. Malondialdehyde of three tissues was increased with an increase in PNP concentration. The expression of four detoxification (cyp450, gst, gpx, hsp70) and one immune-related (mhc II) genes was induced at low PNP concentrations but inhibited at high PNP concentrations in the kidney. In liver, cyp450, hsp70 and mhc II showed similar trend but gst and gpx didn't increase at low PNP concentrations. Our results indicate that the fish possesses the ability to detoxify PNP; however, at high concentrations, PNP still causes serious damage to them. Our data not only help in understanding the ability of L. crocea to detoxify PNP but also should serve as a basis for the study of toxic effects of nitrobenzenes on marine fish. Unlabelled Image • PNP concentration and exposure time were directly proportional to mortality rate. • The activities of CAT and SOD showed synergistic effect in liver and kidney under low PNP stress. • PNP stress led to hepatic cell necrosis and liver function damage. • The immune and secretory functions of the kidney play a major role in resisting PNP stress. • The kidney appeared more sensitive to PNP stress than the liver. [ABSTRACT FROM AUTHOR]