Transcriptomic and histologic analyses preliminarily reveal the immune-metabolic response mechanism to saline-alkaline in large yellow croaker (Larimichthys crocea).

There are large areas of saline-alkaline waters worldwide, the utilization of which would greatly enhance the development of aquaculture productivity. To elucidate the regulatory mechanisms underlying the adaptation of large yellow croaker (Larimichthys crocea) to saline-alkaline water, this study analyzed the growth performance, tissue histology, and gills transcriptome profiles of L. crocea in both seawater (CK) and saline-alkaline water (EX) groups. Growth indices statistics revealed that L. crocea can adapt to saline-alkaline water, with growth performance comparable to that of the CK group. Histological examination revealed partial cellular detachment and structural relaxation in the gills tissue of the EX group, while liver and kidney tissues appeared normal. Transcriptome analysis revealed 3821 differentially expressed genes (DEGs), with 1541 DEGs up-regulated and 2280 DEGs down-regulated. GO enrichment analysis indicated that up-regulated DEGs were enriched in terms related to metabolite production during biological activities, while down-regulated DEGs were associated with terms related to maintaining cellular activities. KEGG enrichment analysis revealed that up-regulated DEGs were enriched in pathways related to the synthesis and metabolism of amino acids and lipids, such as the PPAR signaling pathway and glutathione metabolism. The down-regulated DEGs were predominantly enriched in immune-related signaling pathways, including the Toll-like receptor signaling pathway and NOD-like receptor signaling pathway. Further analysis revealed that genes such as lipoprotein lipase A (lpla), branched-chain amino acid aminotransferase 2 (bcat2), interleukin 8 (il8), interleukin 10 (il10), and interferon regulatory factor 7 (irf7) were involved in the adaptation of L. crocea to saline-alkaline water culture conditions. This study provides a basis for understanding the adaptability of large yellow croaker to saline-alkaline water and lays the foundation for the rational utilization of fishery water resources.

Purification, characterization, and antioxidant ability of polysaccharides from Phascolosoma esculentas.

The polysaccharide was extracted from Phascolosoma esculenta (PEP). Two purified polysaccharides (PEP-1 and PEP-2) were obtained by the column chromatography separation method. The molecular weights of PEP-1 and PEP-2 were 33.6 and 5.7 × 103 kDa, respectively. PEP-1 and PEP-2 had the same monosaccharides composition, but their molar ratios varied. The in vitro antioxidant activity of the PEP, PEP-1, and PEP-2 were investigated by scavenging free radicals like 3-ethylbenzoth-iazoline-6-sulfonic acid (ABTS), •OH, and 2,2-diphenyl-1-picrylhydrazyl (DPPH). Additionally, the in vivo antioxidant activity of PEP-1 was examined using the Caenorhabditis elegans (C. elegans) organism. Results showed that PEP-1 was much more effective than PEP and PEP-2 at scavenging DPPH, •OH, and ABTS radicals. Additionally, PEP-1 strengthened C. elegans' ability to endure oxidative stress. PEP-1 possessed the in vivo antioxidant capacity, including the reactive oxygen species (ROS) content reducing, and protective effect on antioxidant enzyme activities in C. elegans. In summary, PEP, PEP-1, and PEP-2 might have the potential to develop as functional foods and clinical medications.

Evaluation of different agricultural wastes for the production of polysaccharides from Oudemansiella raphanipes and its antioxidant properties.

Oudemansiella raphanipes (OR) is a commercial mushroom which possesses high nutritional value and excellent and unique flavors. In this study, various agricultural wastes were utilized as substitute materials in the low-cost and high-yield production of mycelia biomass and polysaccharides by liquid fermentation. The sawdust, wheat bran, apple pomace, sugarcane, and corn particles were employed to cultivate OR, using the potato dextrose broth as control. Additionally, a preliminary characterization and in vitro antioxidant activities of partial purified OR polysaccharides were investigated. The substrate of sugarcane was suitable for mycelia growth of OR, with high yield of mycelia biomass and polysaccharides content. In vitro antioxidant activity assays demonstrated that OR polysaccharides could effectively scavenge 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 1,1-diphenyl-2-picrylhydrazyl radicals. OR polysaccharides had configuration as revealed by Fourier transform infrared, and was mainly composed of fucose (Fuc), rhamnose (Rha), arabinose (Ara), galactose (Gal), glucose (Glc), xylose (Xyl), mannose (Man), ribose (Rib), and galacturonic acid (Gal-UA), with mass percentages of 3.29%, 0.64%, 1.09%, 16.03%, 72.69%, 0.56%, 3.18%, 0.93%, and 1.59%, respectively. This study may offer support for decreasing the cost of OR polysaccharides production and dealing with these agricultural wastes.

Characterization of the glucocorticoid receptor of large yellow croaker (Larimichthys crocea) and its expression in response to salinity and immune stressors.

Glucocorticoids are steroidal hormones critical to stress responses in vertebrates. To gain further insight into the role of the glucocorticoid receptor (GR) in acute stress responses in teleost fish, the relevant cDNA of large yellow croaker (Larimichthys crocea; LcGR) was cloned using the rapid amplification of cDNA ends (RACE) technique. Multiple alignment of the amino acids (aa) of LcGR and the GR of other teleosts indicated LcGR contained four commonly conserved domains and lacked the 9-aa insert seen in GR1. Phylogenetic analysis of the amino acid sequence revealed that LcGR grouped most closely with the GR2 of other teleosts and can therefore be considered a GR2 subtype. In healthy L. crocea, Lcgr mRNA was found to be expressed at high levels in the gill, brain, and muscle tissue, expressed at intermediate levels in heart and stomach tissue, and expressed at low levels in the kidney, intestine, head kidney, liver, and spleen tissue. The response of L. crocea to acute low-salinity stress was tested, with a significant increase in plasma cortisol concentration after 3 h, peaking after 6 h, and gradually returning to base levels. Regarding changes of Lcgr expression in different body tissues under the stress, there was up-regulation of the Lcgr transcript in the brain, liver, and gill tissues, but not in muscle tissue. Responses to pathogen mimics were also tested. Injection with lipopolysaccharide resulted in Lcgr expression, with an increase-decrease-increase trend in the head kidney. In contrast, a down-regulation of Lcgr expression in the head kidney was observed throughout the experimental period upon injection of polyinosinic:polycytidylic acid, revealing different roles of Lcgr for different types of pathogens. The results offer novel insights about the effects of different stressors on GR gene expression in L. crocea, and can facilitate further investigations into stress responses in other mariculture fish species.

MINDY1 promotes bladder cancer progression by stabilizing YAP.

BACKGROUND: Bladder cancer is one of the most commonly diagnosed urological malignant tumor. The Hippo tumor suppressor pathway is highly conserved in mammals and plays an important role in carcinogenesis. YAP is one of major key effectors of the Hippo pathway. However, the mechanism supporting abnormal YAP expression in bladder cancer remains to be characterized. METHODS: Western blot was used to measure the expression of MINDY1 and YAP, while the YAP target genes were measured by real-time PCR. CCK8 assay was used to detect the cell viability. The xeno-graft tumor model was used for in vivo study. Protein stability assay was used to detect YAP protein degradation. Immuno-precipitation assay was used to detect the interaction domain between MINDY1 and YAP. The ubiquitin-based Immuno-precipitation assays were used to detect the specific ubiquitination manner happened on YAP. RESULTS: In the present study, we identified MINDY1, a DUB enzyme in the motif interacting with ubiquitin-containing novel DUB family, as a bona fide deubiquitylase of YAP in bladder cancer. MINDY1 was shown to interact with, deubiquitylate, and stabilize YAP in a deubiquitylation activity-dependent manner. MINDY1 depletion significantly decreased bladder cancer cell proliferation. The effects induced by MINDY1 depletion could be rescued by further YAP overexpression. Depletion of MINDY1 decreased the YAP protein level and the expression of YAP/TEAD target genes in bladder cancer, including CTGF, ANKRD1 and CYR61. CONCLUSION: In general, our findings establish a previously undocumented catalytic role for MINDY1 as a deubiquitinating enzyme of YAP and provides a possible target for the therapy of bladder cancer.

Suppressive effects of plumbagin on the growth of human bladder cancer cells via PI3K/AKT/mTOR signaling pathways and EMT.

BACKGROUND: Novel chemotherapeutic drugs with good anti-tumor activity are of pressing need for bladder cancer treatment. In this study, plumbagin (PL), a natural plant-derived drug extracted from Chinese herbals, was identified as a promising candidate for human bladder cancer (BCa) chemotherapy. METHODS: The anti-tumor activity of PL was evaluated using a series of in vitro experiments, such as MTT, transwell assay, flow cytometry, quantitative real-time PCR (qRT-PCR) and western blotting. We established xenograft tumors in nude mice by subcutaneous injection with the human bladder cancer T24 cells. RESULTS: The results showed that PL could inhibit the proliferation, migration and survival of BCa cells (T24 and UMUC3 cells) in a time- and dose-dependent way. We found PL promotes the cell cycle arrest and apoptosis by inhibiting PI3K/AKT/mTOR signaling pathway, which inhibits cell proliferation. In vivo, anti-tumor activity of PL was further investigated using a BCa cell xenograft mice model. To simulate clinical chemotherapy, the PL were intravenously injected with a dose of 10 mg/kg for 10 times. Compared with the blank control, the tumor weight in PL treated group decreased significantly from 0.57 ± 0.04 g to 0.21 ± 0.06 g (P < 0.001). CONCLUSIONS: In our study. We found PL inhibits the proliferation of T24 and UMUC3 cells in vivo and in vitro, which may play a role through several downstream effectors of PI3K/AKT/mTOR signaling pathway to promote the cell cycle arrest and apoptosis. Meanwhile, we consider that PL may inhibit the migration of bladder cancer cells via EMT suppression and induce ROS generation to make cell apoptosis. This work screened out a novel chemotherapeutic drug (plumbagin) with relatively good anti-tumor activity, which possessed great potential in BCa chemotherapy.