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Ferroptosis, a nonapoptotic form of cell death marked by iron-dependent peroxidation of phospholipids, is associated with the occurrence and progression of tumors. Erastin, a selective inhibitor of the cystine/glutamate transporter system Xc−, can induce the ferroptosis of cancer cells. Multiple myeloma (MM) has been reported to be insensitive to erastin-induced ferroptosis. However, we found the erastin sensitivity of different MM cells varied widely. Specifically, SLC7A11 abundance determined the sensitivity of MM cells to erastin-induced ferroptosis. MM cells expressing a high SLC7A11 level were more sensitive to erastin-induced ferroptosis than cells expressing a low level of SLC7A11. Moreover, the expression of SLC7A11 gradually increased with the progression of plasma cell dyscrasias. Survival analysis indicated that high levels of SLC7A11 predicted a poor prognosis for MM patients. Knocking down SLC7A11 expression significantly inhibited the proliferation of MM cells and induced ferroptotic cell death. Additionally, we revealed that the long noncoding RNA (lncRNA) SLC7A11-AS1 was a critical regulatory factor of SLC7A11 expression. SLC7A11-AS1 overexpression diminished SLC7A11 levels, leading to the ferroptosis of MM cells. In summary, our data show that heterogeneous SLC7A11 expression affects MM cell sensitivity to ferroptosis, providing a theoretical basis for improving the clinical treatment of MM. [ABSTRACT FROM AUTHOR]

[Display omitted] Lung cancer remains a leading cause of cancer-related mortality worldwide, with a complex molecular pathogenesis that is still not fully understood. Given the challenging task of distinguishing between subtypes of lung cancer (LUAD and LUSC), this study aimed to evaluate the potential of microRNAs (miRNAs) as biomarkers for differentiating these subtypes, by leveraging cancer genomics data. We used bioinformatics analysis for miRNA target prediction. In this regard, a miRNA-mRNA interaction network was defined and pathway analysis was subsequently established. To verify the results of the bioinformatic analysis, we conducted clinical and pre-clinical experiments. We carried out survival analysis, comparing gene expression and mortality rates between two major subtypes of lung cancer, LUSC and LUAD, from 45 lung cancer patients. Additionally, in vitro and in vivo studies were performed using human lung cancer cell lines to alter the expression of miR-21 and miR-326 using plasmid models. Finally, we conducted numerous assays to evaluate cell viability, cell migration and invasion, apoptosis, and cell cycle detection. We observed 102 miRNAs and 1,974 mRNAs as having differential expression in LUAD, whereas 143 miRNAs and 2,821 mRNAs were shown to have such an effect in LUSC. We found that hsa-mir-375 might significantly distinguish LUAD from LUSC, and our clinical validation demonstrated that the level of hsa-mir-375 was significantly higher in LUAD than in LUSC. In terms of finding a diagnostic marker, we discovered that a high level of hsa-mir-21 and/or hsa-mir-326 correlated with a worse prognosis. Our subsequent pre-clinical experiment showed that high expression of hsa-mir-21 and/or hsa-mir-326 significantly increased the aggressiveness of lung cancer and that inhibition of these miRNAs could potentially decrease the tumorigenic behavior of lung cancer cell lines. In conclusion, hsa-mir-21 and hsa-mir-326 might be potential diagnostic biomarkers for lung cancer and hsa-mir-375 might be a potential LUAD and LUSC distinguishing biomarker. [ABSTRACT FROM AUTHOR]

Simple Summary: Esophageal squamous cell carcinoma (ESCC) is a high-incidence cancer in China for which treatment strategies and therapeutic effects are limited. In this study, we established cellular immunotherapy for ESCC using epidermal growth factor receptor (EGFR)-targeting chimeric antigen receptor (CAR)-expressing T cells. The goal of this project was to provide new and effective therapies for ESCC. ESCC is a highly malignant tumor, and its morbidity and mortality in China account for more than 50% of the world's total rates. As effective treatments are lacking, the 5-year survival rate of patients does not exceed 30%. CAR-T-cell-based immunotherapy has emerged as one of the most promising cancer treatments; however, there are relatively fewer reports regarding its application for ESCC. In this study, we conducted large-sample whole-genome sequencing (WGS) and RNA-seq analysis of patients with ESCC from China to examine the feasibility of EGFR-targeting CAR-T cells in the treatment of ESCC. We found much higher levels of EGFR gene amplification and overexpression in tumors than in the normal tissues, indicating that EGFR could be a promising target of CAR-T-cell-based immunotherapy in ESCC. Therefore, we tested EGFR-targeting CAR-T cells for lytic activity against ESCC cells as a model to establish cellular immunotherapy for ESCC. Five types of CAR-T cells targeting EGFR were constructed, two of which, CAR1-T and CAR2-T, showed a strong cytotoxicity against ESCC in in vitro and in vivo experiments. The results of this study suggest that CAR1-T and CAR2-T have the potential to be used for anti-ESCC immunotherapy in clinics. [ABSTRACT FROM AUTHOR]

The purpose of this article is to study the effects and mechanism of miR-4796 in the process of ophiopogon polysaccharide liposome (OPL) regulation of the immune activity of Kupffer cells (KCs). In this study, KCs were used as cell models, and were treated with OPL in different concentrations after being transfected with miR-4796 mimic or miR-4796 inhibitor. Firstly, the secretion of NO and iNOS, phagocytic activity, the expression of surface molecules CD14 and MHC II, apoptosis and ROS secretion were measured by Griess, flow cytometry, fluorescence staining and ELISA. Then, real-time PCR and Western blot were used to measure the expression of TLR4, IKKβ, MyD88 and NF-κB in the TLR4-NF-κB signaling pathway. The results showed that after transfection with miR-4796 mimic, the secretion of NO and iNOS, cell migration, cell phagocytosis and expression levels of CD14 and MHC II in the OPL group were significantly higher than those in the miR-4796 mimic control group (p < 0.05; p < 0.01). In addition, the mRNA and protein expression levels of TLR4, MyD88 and NF-κB were significantly higher than those in miR-4796 mimic control group (p < 0.05; p < 0.01). After transfection with miR-4796 inhibitor, the secretion of NO and iNOS, cell migration, cell phagocytosis, expression of CD14 and MHCII in OPL group were significantly higher than those in the miR-4796 inhibitor control group (p < 0.05; p < 0.01). These results indicated that OPL could regulate the immune activity of KCs by regulating miR-4796 and activating the TLR4-NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Background: The ricefield eel is a freshwater protogynous hermaphrodite fish and has become an important aquaculture species in China. The sex change of ricefield eel is impeding its aquaculture practice, particularly the large-scale artificial breeding. Many studies including transcriptomes of mixed gonadal samples from different individuals have been aimed to elucidate mechanisms underlying the sex change. However, the key physiological factors involved in the initiation of sex change remain to be identified. Results: The present study performed transcriptomic analysis on gonadal samples of different sexual stages obtained through biopsy from the same fish undergoing sex change. A total of 539,764,816 high-quality reads were generated from twelve cDNA libraries of gonadal tissues at female (F), early intersexual (EI), mid-intersexual (MI), and late intersexual (LI) stages of three individual sex-changing fish. Pairwise comparisons between EI and F, MI and EI, and LI and MI identified 886, 319, and 10,767 differentially expressed genes (DEGs), respectively. Realtime quantitative PCR analysis of 12 representative DEGs showed similar expression profiles to those inferred from transcriptome data, suggesting the reliability of RNA-seq data for gene expression analysis. The expression of apoeb, csl2, and enpp2 was dramatically increased and peaked at EI while that of cyp19a1a, wnt4a, fgf16, and foxl2a significantly downregulated from F to EI and remained at very low levels during subsequent development until LI, which suggests that apoeb, csl2, enpp2, cyp19a1a, wnt4a, fgf16, and foxl2a may be closely associated with the initiation of sex change of ricefield eels. Conclusions: Collectively, results of the present study confirmed that the down-regulation of female-related genes, such as cyp19a1a, wnt4a, fgf16, and foxl2a, is important for the sex change of ricefield eels. More importantly, some novel genes, including apoeb, csl2, and enpp2, were shown to be expressed with peak values at EI, which are potentially involved in the initiation of sex change. The present transcriptomic data may provide an important research resource for further unraveling the mechanisms underlying the sex change and testicular development in ricefield eels as well as other teleosts. [ABSTRACT FROM AUTHOR]

Main observation and conclusion: Lithocarpins E—G, featuring a rare naturally‐occurring highly oxygenated tenellone‐macrolide skeleton, were isolated from the culture extract of a marine‐derived fungus Phomopsis lithocarpus FS508. Their structures were fully elucidated by NMR and MS spectroscopic analyses and electronic circular dichroism calculations. Compounds 1—3 were evaluated for their in vitro cytotoxicities against HepG2, MCF‐7, SF‐268, as well as A549 cell lines, among which, compound 1 exhibited inhibitory activity against HepG2 cells with an IC50 value of 6.3 μmol/L. The further mechanistic investigation demonstrated that lithocarpin E could cause the apoptosis of HepG2 cells through activation of p‐ERK, Bax, and caspase‐3 gene expressions. [ABSTRACT FROM AUTHOR]

Whether synthetic genomes can power life has attracted broad interest in the synthetic biology field. Here, we report de novo synthesis of the largest eukaryotic chromosome thus far, synIV , a 1,454,621-bp yeast chromosome resulting from extensive genome streamlining and modification. We developed megachunk assembly combined with a hierarchical integration strategy, which significantly increased the accuracy and flexibility of synthetic chromosome construction. Besides the drastic sequence changes, we further manipulated the 3D structure of synIV to explore spatial gene regulation. Surprisingly, we found few gene expression changes, suggesting that positioning inside the yeast nucleoplasm plays a minor role in gene regulation. Lastly, we tethered synIV to the inner nuclear membrane via its hundreds of loxPsym sites and observed transcriptional repression of the entire chromosome, demonstrating chromosome-wide transcription manipulation without changing the DNA sequences. Our manipulation of the spatial structure of synIV sheds light on higher-order architectural design of the synthetic genomes. [Display omitted] • De novo synthesis of the largest eukaryotic chromosome, synIV • SynIV has 3D structure similar to native chrIV , despite thousands of changes • "Inside-out" intranuclear restructuring of synIV yields minor transcriptional changes • Multipoint tethering synIV to nuclear membrane pervasively represses synIV transcription Zhang et al. synthesized a 1.45-Mb extensively modified yeast chromosome from scratch. The gene position/expression relationship was probed by turning synIV inside-out, exchanging the positions of centromeres and telomeres, with little impact on the transcriptome. When tethering synIV to the inner nuclear membrane, transcription throughout synIV was reduced. [ABSTRACT FROM AUTHOR]

Brain metastasis (BM) is a major complication of lung adenocarcinoma (LAD). An investigation of the pathogenic mechanisms of BM, as well as the identification of appropriate molecular markers, is necessary. The aim of this study was to determine the expression patterns of microRNAs (miRNAs) in LAD with BM, and to investigate the biological role of these miRNAs during tumorigenesis. miRNA array profiles were used to identify BM-associated miRNAs. These miRNAs were independently validated in 155 LAD patients. Several in vivo and in vitro assays were performed to verify the effects of miRNAs on BM. We identified six miRNAs differentially expressed in patients with BM as compared to patients with BM. Of these, miR-4270 and miR-423-3p were further investigated. miR-4270 and miR-423-3p directly targeted MMP19 and P21, respectively, to influence cell viability, migration, and colony formation in vitro. miR-4270 downregulation and miR-423-3p upregulation was associated with an increased risk of BM in LAD patients. Thus, our results suggested that miR-4270 and miR-423-3p might play an important role in BM pathogenesis in LAD patients, and that these miRNAs might be useful prognostic and clinical treatment targets. [ABSTRACT FROM AUTHOR]

Background: Agarwood is a traditional Chinese medicine used as a clinical sedative, carminative, and antiemetic drug. Agarwood is formed in Aquilaria sinensis when A. sinensis trees are threatened by external physical, chemical injury or endophytic fungal irritation. However, the mechanism of agarwood formation via chemical induction remains unclear. In this study, we characterized the transcriptome of different parts of a chemically induced A. sinensis trunk sample with agarwood. The Illumina sequencing platform was used to identify the genes involved in agarwood formation. Methodology/Principal Findings: A five-year-old Aquilaria sinensis treated by formic acid was selected. The white wood part (B1 sample), the transition part between agarwood and white wood (W2 sample), the agarwood part (J3 sample), and the rotten wood part (F5 sample) were collected for transcriptome sequencing. Accordingly, 54,685,634 clean reads, which were assembled into 83,467 unigenes, were obtained with a Q20 value of 97.5%. A total of 50,565 unigenes were annotated using the Nr, Nt, SWISS-PROT, KEGG, COG, and GO databases. In particular, 171,331,352 unigenes were annotated by various pathways, including the sesquiterpenoid (ko00909) and plant–pathogen interaction (ko03040) pathways. These pathways were related to sesquiterpenoid biosynthesis and defensive responses to chemical stimulation. Conclusions/Significance: The transcriptome data of the different parts of the chemically induced A. sinensis trunk provide a rich source of materials for discovering and identifying the genes involved in sesquiterpenoid production and in defensive responses to chemical stimulation. This study is the first to use de novo sequencing and transcriptome assembly for different parts of chemically induced A. sinensis. Results demonstrate that the sesquiterpenoid biosynthesis pathway and WRKY transcription factor play important roles in agarwood formation via chemical induction. The comparative analysis of the transcriptome data of agarwood and A. sinensis lays the foundation for elucidating the mechanism of agarwood formation via chemical induction, and thus, enables future improvements in agarwood quality while protecting endangered wild A. sinensis. [ABSTRACT FROM AUTHOR]

Objective: Except for EGFR gene mutation, there is still lack of predictive factors for gefitinib activity as the second-line treatments for advanced NSCLC with wild-type (WT) EGFR or patients with mutant EGFR but showed poor response. Our purpose was to assess the predictive value of epithelial-mesenchymal transition (EMT) and IGF-1R for gefitinib efficacy as the second-line treatment for NSCLC. Methods: 53 advanced NSCLC patients who accepted gefitinib as the second-line treatment were enrolled in this study. Expression of E-cadherin, vimentin, and IGF-1R was determined by immunohistochemistry. EGFR gene mutation was determined by liquidchip technique. Results: The positive rate of EMT, IGF-1R, and EGFR gene mutation was 54.7%, 58.5%, and 39.6%, respectively. EMT (−) was positively correlated with EGFR gene mutation ( p = .034) and EMT (+) was associated with IGF-1R (+) ( p = .000). EMT (−) was associated with a significantly higher objective response rate (ORR) for all the 53 patients (41.7% vs. 6.9%, p = .024) and showed a higher ORR tendency than EMT (+) in EGFR mutation patients (50.0% vs. 28.6%) and WT EGFR patients (20.0% vs. 4.5%) ( p > .05). EMT (−) showed a significant longer median survival time (MST) than EMT (+) for all 53 patients (8 months vs. 4 months) and WT EGFR patients (6 months vs. 3 months) ( p < .05). IGF-1R (−) showed a higher ORR tendency than IGF-1R (+) in EGFR mutation patients (54% vs. 30%) and WT EGFR patients (18.2% vs. 4.8%) ( p > .05). Conclusion: EMT is correlated with efficacy of gefitinib as the second-line treatment for NSCLC, and combined detection of EMT and IGF-1R may be used as new predictors besides EGFR mutation, especially for patients with WT EGFR. [ABSTRACT FROM AUTHOR]

Long non-coding RNAs (lncRNAs) transcribed extensively from the genome have been proposed to be key regulators of diverse biological processes. However, little is known about the role of lncRNAs in regulation of the cell-cycle G1/S checkpoint following DNA damage, a key step in the maintenance of genomic fidelity. Here we show that growth-arrested DNA damage-inducible gene 7 (gadd7), a DNA damage-inducible lncRNA, regulates the G1/S checkpoint in response to UV irradiation. Interestingly, UV-induced gadd7 directly binds to TAR DNA-binding protein (TDP-43) and interferes with the interaction between TDP-43 and cyclin-dependent kinase 6 (Cdk6) mRNA, resulting in Cdk6 mRNA degradation. These findings demonstrate a role for gadd7 in controlling cell-cycle progression and define a novel mechanism by which lncRNAs modulate mRNA expression at the post-transcriptional level by altering mRNA stability. [ABSTRACT FROM AUTHOR]

The cDNA sequence encoding the prolactin (PRL) gene of the orange-spotted grouper was obtained by randomly sequencing its pituitary cDNA library. The full-length cDNA of orange-spotted grouper PRL consists of 1342 bp. The open reading frame encodes a protein of 212 amino acids, which consists of a putative signal peptide of 24 residues and a mature protein of 188 amino acids. Northern blot analysis identified a single transcript of 1.4 kb in the pituitary gland, indicating perhaps the presence of only one form of PRL in the orange-spotted grouper. The expression of PRL gene in different tissues and during embryonic development of orange-spotted grouper was determined using one-step RT-PCR followed by Southern blot analysis. In addition to the pituitary gland, high levels of PRL gene expression were detected in the olfactory bulb, forebrain, midbrain, hypothalamus, hindbrain, medulla, gill, spleen, and adipose tissue. Very low levels of expression were detected in the kidney, liver, and blood cells. The PRL gene was also expressed in the gonads, with the higher level of expression observed in the stage 3 as compared to stage 2 ovary. During embryonic development, mRNA for PRL was detected at all time points examined except at 2 h 15 min post-fertilization, and its expression peaked at 9 h post-fertilization, at the time of neurulation. Results from the current study suggest that PRL may have important paracrine or autocrine roles in diverse tissue types, including the brain and the gonads, and during embryonic development of the orange-spotted grouper. [Copyright &y& Elsevier]

• Antiserum against ricefield eel Tshb or Tshr generated is of high specificity. • Tsh cells are localized to the inner areas of the adenohypophysis. • Tshb and Tshr expression detected in the ovary and testis at mRNA and protein levels. • Tsh may be produced locally in gonads and potentially play paracrine/autocrine roles. Thyroid stimulating hormone (TSH), a glycoprotein synthesized and secreted from thyrotrophs of the pituitary gland, is composed of a glycoprotein hormone common alpha subunit (CGA) and a specific beta subunit (TSHB). The major biological function of TSH is to stimulate thyroidal follicles to synthesize and secrete thyroid hormones through activating its cognate receptor, the thyroid stimulating hormone receptor (TSHR). In the present study, polyclonal antisera against ricefield eel Tshb and Tshr were generated respectively, and the expression of Tshb and Tshr was examined at mRNA and protein levels. RT-PCR analysis showed that tshb mRNA was expressed mainly in the pituitary as well as in some extrapituitary tissues including the ovary and testis. Tshr mRNA was also expressed in a tissue-specific manner, with transcripts detected in tissues including the kidney, ovary, and testis. The immunoreactive Tshb signals in the pituitary were shown to be localized to the inner areas of adenohypophysis which are close to the neurohypophysis of adult ricefield eels. Tshb-immunoreatvie cells in the pituitary of ricefield eel larvae were firstly observed at hatching. The expression of immunoreactive Tshb and Cga was also detected in ricefield eel ovary and testis together with Tshr. In the ovary, immunoreactive Tshb, Cga, and Tshr were observed in oocytes and granulosa cells. In the testis, immunoreactive Tshb was mainly observed in Sertoli cells while immunoreactive Cga and Tshr were detected in germ cells as well as somatic cells. Results of the present study suggest that Tsh may be synthesized both in the ovary and testis locally, which may play paracrine and/or autocrine roles in gonadal development in ricefield eels. [ABSTRACT FROM AUTHOR]

This study is to investigate the effect of microRNA-1338 (miR-1338) on the activity of Kupffer cells (KCs) and its mechanism regulated by ophiopogon polysaccharide liposome (OPL). KCs was treated with different OPL after transfected with miR-1338 mimic and miR-1338 inhibitor. The secretion of NO and iNOS, the expression of catalase (CAT) and peroxidase (POD), the phagocytic activity, the expression of CD14 and MHC II, the apoptosis and the secretion of ROS were measured. In addition, the expressions of key signal factors TLR4, IKKβ, MyD88 and NF-κB in NF-κB signaling pathway were measured by real-time PCR and Western blot (WB). The results showed that OPL could promote the secretion of iNOS, the expression of POD, the phagocytosis, the mRNA expression of TLR4, MyD88, IKKβ and NF-κB, the protein expression of TLR4 and NF-κB, and inhibit the cell apoptosis and ROS secretion after transfected with miR-1338 mimic. After transfected with miR-1338 inhibitor, OPL could promote the secretion of NO and iNOS, the expression of POD, cell migration, phagocytosis, and inhibit cell apoptosis. Meanwhile, the mRNA expression of TLR4, MyD88, IKKβ and NF-κB and the protein expression of TLR4, MyD88 and NF-κB were promoted. These results suggested that OPL could activate TLR4-NF-κB signaling pathway and thereby improve the activity of KCs by regulating miR-1338. • KCs were treated with OPL after transfected with miR-1338 mimic and inhibitor. • OPL could improve the activity of KCs by regulating miR-1338. • The mechanism of action might be regulation of TLR4-NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]

Small nucleolar RNAs (snoRNAs) are an important group of non-coding RNAs that have been reported to play a key role in the occurrence and development of various cancers. Here we demonstrate that Small nucleolar RNA 42 (SNORA42) enhanced the proliferation and migration of Oesophageal squamous carcinoma cells (ESCC) via the DHX9/p65 axis. Our results found that SNORA42 was significantly upregulated in ESCC cell lines, tissues and serum of ESCC patients. The high expression level of SNORA42 was positively correlated with malignant characteristics and over survival probability of patients with ESCC. Through in vitro and in vivo approaches, we demonstrated that knockdown of SNORA42 significantly impeded ESCC growth and metastasis whereas overexpression of SNORA42 got opposite effects. Mechanically, SNORA42 promoted DHX9 expression by attenuating DHX9 transports into the cytoplasm, to protect DHX9 from being ubiquitinated and degraded. From the KEGG analysis of Next-Generation Sequencing, the NF-κB pathway was one of the most regulated pathways by SNORA42. SNORA42 enhanced phosphorylation of p65 and this effect could be reversed by NF-κB inhibitor, BAY11–7082. Moreover, SNORA42 activated NF-κB signaling through promoting the transcriptional co-activator DHX9 interacted with p-p65, inducing NF-κB downstream gene expression. In summary, our study highlights the potential of SNORA42 is up-regulated in ESCC and promotes ESCC development partly via interacting with DHX9 and triggering the DHX9/p65 axis. • The paper found that SNORA42 was upregulated in ESCC cell lines, tissues and serum. • The high expression of SNORA42 was positively correlated with malignant characteristics and over survival. • In vitro and in vivo approaches indicate that SNORA42 promotes ESCC growth and metastasis. • Mechanically, SNORA42 promoted DHX9 expression by attenuating DHX9 transports into the cytoplasm, to protect DHX9 from being ubiquitinated and degraded. • Moreover, SNORA42 activated NF-κB signaling through promoting DHX9 interacted with p-p65, inducing NF-κB downstream gene. • In summary, our study highlights the potential of SNORA42 is up-regulated in ESCC and promotes ESCC development partly via interacting with DHX9 and triggering the DHX9/p65 axis. [ABSTRACT FROM AUTHOR]

Transmissible gastroenteritis virus (TGEV) is an important pathogen capable of altering the expression profile of cellular miRNA. In this study, the potential of Polygonum c illinerve polysaccharide (PCP) to treat TGEV-infected piglets was evaluated through in vivo experiments. High-throughput sequencing technology was employed to identify 9 up-regulated and 17 down-regulated miRNAs during PCP-mediated inhibition of TGEV infection in PK15 cells. Additionally, miR-181 was found to be associated with target genes of key proteins in the apoptosis pathway. PK15 cells were treated with various concentrations of PCP following transfection with miR-181 mimic or inhibitor. Real-time PCR assessed the impact on TGEV replication, while electron microscopy (TEM) and Hoechst fluorescence staining evaluated cellular functionality. Western blot analysis was utilized to assess the expression of key signaling factors—cytochrome C (cyt C), caspase 9, and P53—in the apoptotic signaling pathway. The results showed that compared with the control group, 250 μg/mL PCP significantly inhibited TGEV gRNA replication and gene N expression (P < 0.01). Microscopic examination revealed uniform cell morphology and fewer floating cells in PCP-treated groups (250 and 125 μg/mL). TEM analysis showed no typical virus structure in the 250 μg/mL PCP group, and apoptosis staining indicated a significant reduction in apoptotic cells at this concentration. Furthermore, PCP may inhibit TGEV-induced apoptosis via the Caspase-dependent mitochondrial pathway following miR-181 transfection. These findings provide a theoretical basis for further exploration into the mechanism of PCP's anti-TGEV properties. • PCP possessed better anti-TGEV effects in piglets. • PCP significantly inhibited TGEV replication by regulating miR-181. • PCP inhibited TGEV-induced apoptosis by mitochondrial pathway by regulating miR-181. [ABSTRACT FROM AUTHOR]

Aromatase (encoded by Cyp19a1) in the ovarian follicular cells catalyzes the production of estradiol from testosterone, which plays important roles in the ovarian development of vertebrates. In the present study, the interaction of Dmrt1, Foxl2, and Nr5a1a on the regulation of cyp19a1a transcription in ovarian follicles was examined in a teleost, the ricefield eel Monopterus albus. The expression of dmrt1a , foxl2 , and nr5a1a was detected in ovarian follicular cells together with cyp19a1a at the mRNA and/or protein levels. Sequence analysis identified one conserved Foxo binding site in the proximal promoter region of ricefield eel cyp19a1a. Transient transfection assay showed that Foxl2 may bind to the conserved Foxo site to activate cyp19a1a transcription and act synergistically with Nr5a1a. Mutation of either the conserved Nr5a1 site or Foxo site abolished or significantly decreased the synergistic effects of Nr5a1a and Foxl2 on cyp19a1a transcription. The sequence between Region III and I-box of Nr5a1a was critical to this synergistic effect. Dmrt1a modulated the Foxl2- and Nr5a1a-induced activation of cyp19a1a transcription and their synergistic effects in a biphasic manner, with inhibitory roles observed at lower doses (10–50 ng) but release of the inhibition or even potentiating effects observed at higher doses (100–200 ng). Collectively, data of the present study suggest that the interaction of Dmrt1a, Foxl2, and Nr5a1a in the ovarian follicular cells may facilitate the adequate expression of cyp19a1a and the production of estradiol, and contribute to the development and maturation of ovarian follicles in ricefield eels and other vertebrates as well. • cyp19a1a , nr5a1a , foxl2 , and dmrt1a were expressed in follicular cells. • Foxl2 up-regulated cyp19a1a transcription via the conserved Foxo site. • Nr5a1a and Foxl2 bound to cyp19a1a promoter to exert their full synergism. • The sequence between Region III and I-box of Nr5a1a was key to the synergism. • Dmrt1a abolished the synergism of Nr5a1a and Foxl2 on c yp19a1a transcription. [ABSTRACT FROM AUTHOR]

Abstract: Sox8 and Sox10 are members of group E Sox proteins involved in a wide range of developmental processes including sex determination and neurogenesis in vertebrates. The orange-spotted grouper sox8a and sox10a homologues were isolated and characterized in the present study. Both sox8a and sox10a genes contain three exons and two introns, and encode putative proteins with typical structures of group E Sox. Sox8a was expressed in diverse tissues including the central nervous system and some peripheral tissues. In contrast, sox10a mRNA was detected primarily in the central nervous system. During embryogenesis, sox8a mRNA seemed to be de novo synthesized in the embryos from otic vesicle stage. However, sox10a mRNA was only detectable in juvenile fish 35days post hatching and thereafter. The mRNA levels of sox8a in the gonads were not significantly different among ovarian developmental stages but increased in the testis. In vitro transfection assays showed that the Sox10a but not Sox8a up-regulated cyp19a1a promoter activities. Taken together, these results suggested that the sox8a may play roles in diverse tissues and during embryogenesis, whereas sox10a may be mainly involved in the neural regulation of juvenile and adult fish, and that certain Sox homologues may regulate the orange-spotted grouper cyp19a1a promoter. [Copyright &y& Elsevier]

Abstract: CYP19A1 in the brain and pituitary of vertebrates is important for reproductive and non-reproductive processes. In teleosts, it is broadly accepted that estradiol (E2) up-regulates cyp19a1b gene via a positive autoregulatory loop. Our present study, however, showed that E2 did not up-regulate ricefield eel cyp19a1b in the hypothalamus and pituitary, whereas dihydrotestosterone (DHT) or testosterone (T) stimulated cyp19a1b expression only in the pituitary. Two tissue-specific promoters, namely promoter I and II directing the expression in the brain and pituitary respectively, were identified. Promoter I contained a non-consensus estrogen response element (ERE), and consequently did not respond to E2. Promoter II contained an androgen response element (ARE) and consequently responded to DHT. Taken together, these results demonstrated a novel steroidal regulation of cyp19a1b gene expression and an alternative usage of tissue-specific cyp19a1b promoters in the brain and pituitary of a teleost species, the ricefield eel. [Copyright &y& Elsevier]

Abstract: Sox genes, a family of genes related to the mammalian sex-determining region Y (SRY) gene, are found throughout the animal kingdom, and involved in diverse developmental processes including sex determination and neurogenesis. Previously, we have identified one sox11 homologue, sox11b, from the ovary of the orange-spotted grouper. In the present study, another sox11 homologue, sox11a, was cloned from the brain. The orange-spotted grouper Sox11a contained the signature features of mammalian SOX11 homologues except the Pro–Glu rich region, was clustered with Sox11a homologues of other teleosts in the phylogenetic tree, and shared higher homologies with Sox11 of other species than the duplicated copy Sox11b. Interestingly, significant conservation was observed in the 3′UTR of sox11a but not sox11b transcripts when compared with mammalian Sox11 homologues. The expression of sox11a mRNA was detected in a wide range of tissues, with higher abundances in the central nervous system. During embryogenesis and larval development, the expression of sox11a mRNA remained at considerably high levels at all stages examined, from newly fertilized eggs, through organogenesis, to the larvae 18days posthatching. Together, these results indicated that the orange-spotted grouper sox11a was evolutionarily more conserved than sox11b, and may play important roles in neurogenesis, embryogenesis, and larval development. [ABSTRACT FROM AUTHOR]

• Specific antisera were generated for ricefield eel Esr2a and Esr2b. • Esr2a expressed in the adenohypophysis while Esr2b in the neurohypophysis. • Esr2a localized to Prl and Gh cells but not Lh and Fsh cells. • E2 stimulated prl and lhb gene expression in pituitary cells. • E2 had no effects on gh , fshb , and gnrhr2 gene expression in pituitary cells. Estrogens play important regulatory roles in the pituitary of vertebrates. Two forms of estrogen receptor 2 (Esr2), namely Esr2a and Esr2b, are identified in teleosts, but their differential roles remain to be fully elucidated. In the present study, expression and potential functional roles of Esr2a and Esr2b were characterized in ricefield eels. esr2a and esr2b mRNA were broadly distributed in tissues, with high levels observed in the brain, pituitary, and gonads. In order to examine the cellular localization of Esr2a and Esr2b in the pituitary, specific antisera against ricefield eel Esr2a and Esr2b were generated, respectively. Interestingly, immunohistochemistry and Western blot analysis revealed that Esr2a and Esr2b were differentially distributed in the pituitary, with the former localized to the adenohypophysis while the latter to the neurohypophysis. Dual fluorescent immunostaining showed that immunoreactive Esr2a was present in Gh and Prl cells, but not in Lh and Fsh cells. Estradiol (E 2) stimulated lhb and prl gene expression in dispersed pituitary cells of intersexual ricefield eels, but had no effects on gh , fshb , and gnrhr2 gene expression and Gh release. Results of the present study are helpful for further understanding the roles and mechanisms of estrogen signals in the pituitary. [ABSTRACT FROM AUTHOR]