Search

Aim: This study aimed to determine the effects of iRoot BP Plus on receptor activator of nuclear factor‐κB ligand (RANKL)‐induced osteoclastogenesis in vitro and inflammation‐mediated bone resorption in vivo and investigated the underlying molecular mechanisms. Methodology: CCK‐8 was performed to test cell viability in RANKL‐induced RAW 264.7 cells and BMDMs in response to iRoot BP Plus. The effect of iRoot BP Plus on osteoclastogenesis was determined using TRAP staining and phalloidin staining, respectively. Pit formation assay was conducted to measure osteoclast resorptive capacity. Western blot and qPCR were performed to examine osteoclast‐related proteins and gene expression, respectively. Western blot was also used to investigate the signalling pathways involved. For in vivo experiments, an LPS‐induced mouse calvarial bone resorption model was established to analyse the effect of iRoot BP Plus on bone resorption (n = 6 per group). At 7 days, mouse calvaria were collected and prepared for histological analysis. Results: We identified that iRoot BP Plus extracts significantly attenuated RANKL‐induced osteoclastogenesis, reduced sealing zone formation, restrained osteolytic capacity and decreased osteoclast‐specific gene expression (p <.01). Mechanistically, iRoot BP Plus extracts reduced TRAF6 via proteasomal degradation, then suppressed the phosphorylation of mitogen‐activated protein kinases (MAPKs), blocked the nuclear translocation of c‐Fos and diminished nuclear factor‐κB (NF‐κB) p65 and NFATc1 accumulation. Consistent with the in vitro results, iRoot BP Plus extracts attenuated osteoclast activity thus protecting against inflammatory bone resorption in vivo (p <.05), which was accompanied by a suppression of TRAF6, c‐Fos, NFATc1 and cathepsin K expression. Conclusion: These findings provide valuable insights into the signalling mechanisms underlying nanoparticulate bioceramic putty‐mediated bone homeostasis. [ABSTRACT FROM AUTHOR]

The light/dark cycle, known as the photoperiod, plays a crucial role in influencing various physiological activities in fish, such as growth, feeding and reproduction. However, the underlying mechanisms of this influence are not fully understood. This study focuses on exploring the impact of different light regimes (LD: 12 h of light and 12 h of darkness; LL: 24 h of light and 0 h of darkness; DD: 0 h of light and 24 h of darkness) on the expression of clock genes (LcClocka, LcClockb, LcBmal, LcPer1, LcPer2) and the secretion of hormones (melatonin, GnRH, NPY) in the large yellow croaker, Larimichthys crocea. Real-time quantitative PCR (RT-qPCR) and enzyme-linked immunosorbent assays were utilized to assess how photoperiod variations affect clock gene expression and hormone secretion. The results indicate that changes in photoperiod can disrupt the rhythmic patterns of clock genes, leading to phase shifts and decreased expression. Particularly under LL conditions, the pineal LcClocka, LcBmal and LcPer1 genes lose their rhythmicity, while LcClockb and LcPer2 genes exhibit phase shifts, highlighting the importance of dark phase entrainment for maintaining rhythmicity. Additionally, altered photoperiod affects the neuroendocrine system of L. crocea. In comparison to the LD condition, LL and DD treatments showed a phase delay of GnRH secretion and an acceleration of NPY synthesis. These findings provide valuable insights into the regulatory patterns of circadian rhythms in fish and may contribute to optimizing the light environment in the L. crocea farming industry. [ABSTRACT FROM AUTHOR]

Chinese narcissus (Narcissus tazetta var. chinensis cv. 'Jinzhanyintai') is one of the 10 most famous traditional flowers of China, having a beautiful and highly ornamental flower with a rich fragrance. However, the flower longevity affects its commercial appeal. While petal senescence in Narcissus is ethylene-independent and abscisic acid-dependent, the regulatory mechanism has yet to be determined. In this study, we identified a R2R3-MYB gene (NtMYB1) from Narcissus tazetta and generated oeNtMYB1 and Ntmyb1 RNA interference mutants in Narcissus as well as an oeNtMYB1 construct in Arabidopsis. Overexpressing NtMYB1 in Narcissus or Arabidopsis led to premature leaf yellowing, an elevated level of total carotenoid, a reduced level of chlorophyll b , and a decrease in photosystem II fluorescence (F v/ F m). A dual-luciferase assay and chromatin immunoprecipitation–quantitative PCR revealed that NtMYB1 directly binds to the promoter of NtNCED1 or NtNCED2 and activates NtNCED1/2 gene expression both in vitro and in vivo. Moreover, overexpressing NtMYB1 accelerated abscisic acid biosynthesis, up-regulated the content of zeatin and abscisic acid, and down-regulated the level of β-carotene and gibberellin A1, leading to petal senescence and leaf yellowing in Narcissus. This study revealed a regulatory process that is fundamentally different between non-photosynthetic organs and leaves. [ABSTRACT FROM AUTHOR]

The skin of Trachinotus ovatus is a crucial component of the mucosal immune system and serves as the primary site of infection by Cryptocaryon irritans. In order to investigate the significant role of skin in C. irritans infection, a comprehensive transcriptome analysis was conducted on skin tissues from the infection group, infection-adjacent group, and infection group compared with the infection-adjacent group (ATT_vs_PER, ADJ_vs_PER, ATT_vs_ADJ). This study identified differentially expressed long non-coding RNAs (DE lncRNAs), microRNAs (DE miRNAs), and differentially expressed genes (DEGs). The prediction of lncRNA target genes was accomplished by utilizing positional relationship (co-location) and expression correlation (co-expression) with protein-coding genes. Subsequently, functional enrichment analysis was conducted on the target genes of differentially expressed lncRNAs, revealing their involvement in signaling pathways such as tight junction, MAPK, and cell adhesion molecules. This study describes the regulatory network of lncRNA-miRNA-mRNA in T. ovatus skin tissue infected with C. irritans. Functional prediction analysis showed that differentially expressed lncRNA and miRNA may regulate the expression of immune genes such as interleukin-8 (il8) to resist the infection of C. irritans. Conducting additional research on these non-coding RNAs will facilitate a deeper understanding of their immune regulatory function in T. ovatus during C. irritans infection. The study of non-coding RNA in this study laid a foundation for revealing the molecular mechanism of the immune system of T. ovatus to respond to the infection of C. irritans. It provided a choice for the molecular breeding of Trachinotus ovatus against C. irritans. [ABSTRACT FROM AUTHOR]

Background: To analyze the fibroblasts subtypes in the gingival tissues of healthy controls, gingivitis and periodontitis patients, as well as the effects of interaction between subtypes on alveolar bone destruction.Methods: Gingival tissues were divided into three groups according to clinical and radiographic examination, and the immunostaining of EDA+FN was assessed. Fibroblasts from gingiva developed colony formation units (CFUs) and induced Trap+MNCs. The expression of osteoclastogenesis-related genes was assessed by real-time PCR. Variances in the gene profiles of CFUs were identified by principal component analysis, and cluster analysis divided CFUs into subtypes. The induction of Trap+MNCs and gene expression were compared among individual or cocultured subtypes. The fibroblast subtypes exerted critical effect on Trap+MNCs formation were selected and edited by CRISPR/Cas to investigate the influence on osteoclastogenesis in the periodontitis in mice.Results: Most periodontitis samples exhibited intensive EDA+FN staining (P < 0.05), and these fibroblasts also induced most Trap+MNCs among three groups; consistently, fibroblasts from periodontitis highly expressed genes facilitating osteoclastogenesis. According to gene profiles and osteoclastogenic induction, four clusters of CFUs were identified. The proportion of clusters was significantly different (P < 0.05) among three groups, and their interaction influenced osteoclastogenic induction. Although Cluster 4 induced less osteoclasts, it enhanced the effects of Clusters 1 and 3 on Trap+MNCs formation (P < 0.05). EDA knockout in Cluster 4 abrogated this promotion (P < 0.05), and decreased osteoclasts and alveolar bone destruction in experimental periodontitis (P < 0.05).Conclusion: Heterogeneous fibroblast subtypes affect the switch or development of periodontitis. A subtype (Cluster 4) played important role during alveolar bone destruction, by regulating other subtypes via EDA+FN paracrine. [ABSTRACT FROM AUTHOR]

The sex determination and differentiation process of economically important crustaceans have been regarded as the focus of aquaculture for a long time, because of the sex-related weight differences. Neocaridina denticulata sinensis makes a suitable animal model for studying crustaceans because it can reproduce many times under artificial control and has a short reproductive cycle. Male and female sex characteristics of the adult rice shrimp Neocaridina d. sinensis are morphologically obvious, but not in embryos and juvenile stages. At present, sex-specific DNA markers have not yet been developed. To produce a reliable molecular marker for sex in Neocaridina and to investigate molecular sex differentiation, we therefore focused on identifying sex-specific transcriptomic differences. In this study, we found three sex-specific expression genes, NDM , Sushi , and NDF , after screening a large number of transcriptome data. NDM and Sushi are male-specific expression genes, and NDF is a female-specific expression gene. Semi-quantitative RT-PCR analysis showed that NDM and NDF can act as molecular markers for the sex identification of Neocaridina in different developmental stages, especially sex identification for embryos and juveniles with the same morphological characteristics. However, Sushi can only act as a molecular marker for the sex identification of Neocaridina in adult stages. Furthermore, in situ hybridization showed that a strong positive signal of NDM was detected in the male testis. At the same time, we explored the relationship between these three genes and sex differentiation. The results of RNA interference treatment show that knockdown of nd-IAG (Neocaridina denticulata sinensis insulin-like androgenic gland hormone) can change the expression of NDM and NDF. On the basis of the expression of the male-specific gene NDM and the female-specific gene NDF , we developed a molecular test that for the first time allows the unambiguous sex determination of Neocaridina samples lacking external sex-specific features from juvenile stages onward. Résumé: A cause des différences de poids en fonction du sexe, la détermination du sexe et ses mécanismes de différenciation chez les crustacés économiquement importants ont été pendant longtemps au centre de l'attention en aquaculture. Neocaridina denticulata sinensis est un bon modèle pour étudier les crustacés car elle peut se reproduire plusieurs fois sous contrôle artificiel et a un cycle reproducteur court. Les caractéristiques sexuelles mâle et femelle de la crevette adulte Neocaridina d. sinensis sont morphologiquement évidentes, mais pas chez les embryons ou les stades juvéniles. De nos jours, des marqueurs AND spécifiques du sexe n'ont pas encore été développés. Afin de produire un marqueur moléculaire fiable pour le sexe chez Neocaridina et pour explorer la différenciation moléculaire du sexe, nous nous sommes concentrés sur l'identification des différences transcriptomiques spécifiques du sexe. Dans cette étude, après un grand nombre de criblage des données du transcriptome, nous avons trouvé trois gènes d'expression spécifique du sexe, NDM , Sushi , et NDF. NDM et Sushi sont des gènes d'expression spécifiques mâles, et NDF est spécifique femelle. Des analyses par RT-PCR semi-quantitative ont montré que NDM et NDF peuvent servir comme marqueurs moléculaires pour l'identification du sexe de Neocaridina chez différents stades de développement, pour des embryons et des juvéniles avec les mêmes caractéristiques morphologiques. Cependant, Sushi peut seulement server comme marqueur moléculaire pour l'identification du sexe de Neocaridina chez les adultes. De plus, l'hybridation in situ a montré qu'un signal positif fort pour NDM était détecté dans le testicule mâle. En même temps, nous avons exploré l'interrelation entre ces trois gènes et la différenciation sexuelle. Les résultats du traitement de l'interférence par ARN montrent que l'extinction de nd-IAG (hormone de Neocaridina denticulata sinensis semblable à l'insuline de la glande androgène) peut modifier l'expression de NDM et NDF. Sur la base de l'expression du gène spécifique mâle NDM et du gène spécifique femelle NDF , nous avons développé un test moléculaire qui, pour la première fois, permet une détermination du sexe sans ambiguïté sur des échantillons de Neocaridina sans caractère sexuel externe à partir des stades juvéniles. [ABSTRACT FROM AUTHOR]

Background: The P2Y12 receptor is a kind of purinoceptor that is engaged in platelet aggregation, and P2Y12 inhibitors have been used in clinical antithrombotic therapy. The P2Y12 receptor in microglia induces interleukin-1β (IL-1β) expression, which is a key mediator of depression in the brain. Although peripheral P2Y12 is involved in neuropathic pain, whether P2Y12 expression in the medial prefrontal cortex (mPFC) is associated with comorbidities of visceral pain and depression remains unclear. Accumulating evidence suggests that electroacupuncture (EA) is effective in treating inflammatory bowel disease (IBD), but its mechanism is unknown. This study aimed to determine whether P2Y12 expression in the mPFC is associated with comorbidities of visceral pain and depression in IBD and whether EA treats IBD by targeting the P2Y12 receptor. Methods: We used 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced IBD mice. P2Y12 short hairpin RNA (shRNA) was stereotaxically injected into the bilateral mPFC. EA was performed on bilateral "Dachangshu" (BL25) acupoints once a day for 7 days. Von Frey filaments and colorectal distension were used to detect the mechanical pain threshold and visceral pain sensitivity. The sucrose preference test, tail suspension test and forced swimming test were used to evaluate depression in mice. Western blotting was used to test the expression of P2Y12 and IL-1β. Immunofluorescence staining was used to assess microglial activity. Results: We found that IBD mice presented visceral pain and depression associated with increased P2Y12 expression in the mPFC. P2Y12 shRNA significantly attenuated visceral pain and depression in IBD mice. P2Y12 shRNA significantly downregulated IL-1β expression and inhibited the activation of microglia in the mPFC of IBD mice. Meanwhile, EA played a similar role of P2Y12 shRNA. EA significantly downregulated P2Y12 expression, weakened the activation of microglia, and then inhibited IL-1β expression in the mPFC, thus relieving visceral pain and depression in IBD mice. Conclusion: The present study provided new ideas that the P2Y12 receptor in the mPFC could be a new target for the treatment of comorbid visceral pain and depression by EA. This may not only deepen our understanding of the analgesic and antidepressant mechanisms of EA but also promote the application of EA to treat IBD. [ABSTRACT FROM AUTHOR]

The sea cucumber Apostichopus japonicus is an economically important marine species in China, and understanding the mechanisms underlying its gonad development is crucial for successful reproduction and breeding. In this study, we performed transcriptome comparisons and analyses of A. japonicus gonadal and non-gonadal tissues to identify genes and molecular pathways associated with gonadal development. We also supplemented the annotation of the A. japonicus genome. Collectively, results revealed a total of 941 ovary-specific genes and 2499 testis-specific genes through different expression analysis and WGCNA analysis. The most enriched pathways in ovary and testis were "DNA replication" and "purine metabolism", respectively. Additionally, we identified key candidate gene modules that control gonad development and germ cell maturation, with CDT1 and DYNC2LI1 serving as hub genes. Our findings provide important insights into the gonadal development system of A. japonicus and offer valuable references for further research on reproductive biology in this marine invertebrate species. [Display omitted] • Comparative analyses of gene expression in sea cucumber gonadal and non-gonadal tissues • Key candidate gene modules specifically expressed by females and males gonad were screened. • The ovarian module with CDT1 as the core gene may affect ovarian development through the pre-replication complex. [ABSTRACT FROM AUTHOR]

Aestivation is a state of dormancy in diverse animal species characterized by a depressed metabolism. High temperatures in the summer trigger aestivation in the adult sea cucumber, Apostichopus japonicus. In the present study, the glycolytic suppression in aestivating A. japonicus was verified by metabolite quantification and activity and a gene expression analysis of rate-limiting enzymes (hexokinase and pyruvate kinase). Significant decreases in glucose concentrations were discovered in the respiration tree ( p < 0.01), intestine ( p < 0.05) and muscle ( p < 0.01) of aestivating A. japonicus; likewise, over this period, pyruvic acid concentrations significantly decreased in the respiration tree ( p < 0.05) and intestine ( p < 0.01), but were unchanged in muscle. Enzymatic activity variation of hexokinase ( Aj-HK) and pyruvate kinase ( Aj-PK) in aestivating A. japonicus revealed that Aj-HK remained at high levels in the respiration tree and muscle but significantly decreased in intestine ( p < 0.01); the activity of Aj-PK also declined significantly in intestine ( p < 0.01), whereas it increased significantly in the respiration tree ( p < 0.05) and muscle ( p < 0.01). The gene sequences of Aj-HK and Aj-PK were identified, and their transcriptional quantification revealed that the expression of Aj-HK was depressed in all of the examined tissues, whereas Aj-PK expression was depressed in muscle only. These results suggest that glycolytic suppression occurred in aestivating A. japonicus, especially in the intestine, and transcriptional and post-transcriptional modifications are involved in the regulation of glycolytic suppression in this marine species. [ABSTRACT FROM AUTHOR]

Cathepsin L, a lysosomal endopeptidase, has been noted for its involvement in the innate immune response in invertebrates. Here, the cathepsin L cDNA of the sea cucumber Apostichopus japonicus ( Aj CatL) is identified from an EST library and then cloned by the rapid amplification of the cDNA ends (RACE) PCR. The full-length cDNA is 1678 bp long containing an open reading frame (ORF) of 1002 bp, an 80 bp 5′ UTR and a 599 bp 3′ UTR. The cDNA encodes 333 amino acid residues with a predicted molecular mass of 37.07 kDa and a theoretical isoelectric point (pI) of 5.01. The full-length Aj CatL contains three active sites of eukaryotic thiol (cysteine) protease at positions 133–144, 278–288 and 295–314. Analysis of the predicted tertiary structure of prepro-CatL (17–333 aa) and mature-CatL (116–333 aa) reveals that the propeptide region (17–115 aa) blocks access to the substrate-binding cleft. Phylogenetic analysis shows that the Aj CatL is clustered together with two other CatLs from Strongylocentrotus purpuratus . The enzymatic activity of Aj CatL was verified using a substrate hydrolyzing assay with recombinant m Aj CatL. Further analysis of real time-PCR demonstrates that the expression of Aj CatL mRNA is significantly up-regulated in the coelomocytes in cases of infection with the common bacterial pathogen, Vibrio splendidus . This suggests that the Aj CatL is likely to be involved in the immune response. [ABSTRACT FROM AUTHOR]

In mammals, the type II melanoma antigen (MAGE) protein family is constituted by at least ten closely related members, but our understanding of their function in the developing nervous system remains poor. To systematically study the expression pattern of type II MAGE genes during neurogenesis, we employed mouse embryonic carcinoma P19 cells as an in vitro model for neural differentiation by retinoic acid (RA) induction. The expression of type II MAGE genes was investigated under distinct steps of differentiation by a comparative ΔΔ C paradigm of real-time quantitative reverse-transcription PCR (qRT-PCR). The relative levels of each gene expression at various steps of differentiation were expressed as a fold change compared with that in RA-untreated P19 cells. The results revealed that: (1) the expression of MAGE- E1, E2, and Necdin transcripts was steadily increased, and the relative levels of MAGE- D1, D2, D3, F1, G1, and H1 mRNA were fluctuantly elevated after the RA-treatment at embryoid body and neural stages; (2) during RA-treatment and subsequent differentiation, the expression of MAGE- L2 mRNA was decreased. Therefore, our results suggested that MAGE-D1, D2, D3, E1, E2, F1, G1, H1, and Necdin might be involved in the early process of neurogenesis, and MAGE-L2 connected with maintenance of pluripotency of stem cells. These studies may present some clues for a better understanding of the fundamental aspects of type II MAGE genes during neurogenesis. [ABSTRACT FROM AUTHOR]

Sea cucumber Apostichopus japonicus displays the typical circadian rhythms. This present study investigated the molecular regulation of clock genes, as well as monoamines and melatonin, in multiple tissues of A. japonicus , responding to the photoperiod. In order to determine their pivotal role in circadian rhythms, the crucial clock genes, namely AjClock , AjArnt1 , AjCry1 , and AjTimeless , were identified and a comprehensive analysis of their expressions across various tissues in adult A. japonicus was conducted, revealing the potential existence of central and peripheral oscillators. Results demonstrated that the tissues of polian vesicle and nerve ring exhibited significant clock gene expression associated with the orchestration of circadian regulation, and that environmental light fluctuations exerted influence on the expression of these clock genes. However, a number of genes, such as AjArnt1 and AjCry1 , maintained their circadian rhythmicity even under continuous light conditions. Moreover, we further investigated the circadian patterns of melatonin (MT), serotonin (5-HT), and dopamine (DA) secretion in A. japonicus , data that underscored the tissue-specific regulatory differences and the inherent adaptability to dynamic light environments. Collectively, these findings will provide the molecular mechanisms controlling the circadian rhythm in echinoderms and the candidate tissues playing the role of central oscillators in sea cucumbers. [Display omitted] • The molecular circadian system was identified in Apostichopus japonicus. • The polian vesicle and nerve ring were determined with roles as circadian oscillators. • A. japonicus circadian system exhibits stability. • The rhythmicity of molecular circadian system can be affected by photoperiods. [ABSTRACT FROM AUTHOR]