Aqueous Zn‐ion batteries have been considered as promising alternatives to Li‐ion batteries due to their abundant reserves, low price, and high safety. However, Zn anode shows poor reversibility and ...cycling stability in most conventional aqueous electrolytes. Here, a new type of aqueous Zn‐ion electrolyte based on ZnCl2–acetamide deep eutectic solvent with both environmental and economic friendliness has been prepared. The water molecule introduced in the “water‐in‐deep eutectic solvent” electrolyte could reduce the Zn2+ desolvation energy barrier by regulating Zn2+ solvation structure to promote uniform Zn nucleation. Zn anode shows improved electrochemical performance (≈98% Coulombic efficiency over 1000 cycles) in the electrolyte whose molar ratio of ZnCl2:acetamide:H2O is 1:3:1. The assembled full battery composed of phenazine cathode and Zn anode could stably cycle over 10 000 cycles with a high capacity retention of 85.7%. Overall, this work offers new insights into exploring new green electrolyte systems for Zn‐ion batteries.
The “water‐in‐deep eutectic solvent” electrolyte based on ZnCl2, acetamide, and water with both environmental and economic friendliness can induce uniform Zn nucleation by modulating Zn2+ solvation structure to achieve the fast reaction kinetics on the interface between the electrolyte and Zn metal, accounting for the improved reversibility and cycling stability of the Zn anode.
Age-related osteoporosis, a high-prevalence disease in the aged population, is generally attributed to the excessive activity of osteoclasts. Most approved drugs treat osteoporosis by inhibition of ...osteoclasts. Although in vivo studies have shown that alpha-ketoglutarate (AKG), an intermediate in the TCA cycle, can ameliorate age-related osteoporosis, the effects of AKG on osteoclastogenesis and the underlying mechanism of its action have not been studied yet. Here, we showed that the elevation of intracellular AKG levels by supplementing dimethyl AKG (DM-AKG, a cell-permeable derivative of AKG) inhibits the receptor activator of NF-κB ligand (RANKL)-induced osteoclasts differentiation from primary bone marrow-derived macrophages (BMMs) and RAW264.7 cells in vitro. We further found that DM-AKG treatment suppresses NF-κB signaling and oxidative phosphorylation (OXPHOS) during RANKL-induced osteoclastogenesis in RAW264.7 cells. Interestingly, dimethyl oxalylglycine (DMOG), an AKG competitive inhibitor of AKG-dependent prolyl hydroxylases (PHDs), antagonizes the suppression of the RANKL-activated NF-κB signaling pathway caused by DM-AKG treatment. Furthermore, blocked PHD1 expression (also known as EglN2), instead of PHD2 or PHD3, was confirmed to reverse the DM-AKG treatment-induced suppression of the RANKL-activated NF-κB signaling pathway. Accordingly, blocked PHD1 expression antagonized the inhibitory effects of DM-AKG on osteoclastogenesis. Together, our finding suggests that the elevation of intracellular AKG levels inhibits osteoclastogenesis by suppressing RANKL-activated NF-κB signaling in a PHD1-dependent manner, which may provide a novel nutritional strategy for osteoporosis treatment.
The small yellow croaker (Larimichthys polyactis) is one of the most highly cultured fish in China, contributing considerably to the country's economy. However, conditions experienced under ...high-density culture make the fish susceptible to hypoxia, which impacts growth and consequently product quality. To understand the effects of hypoxia stress on physiological and biochemical responses of L. polyactis, we subjected the fish to hypoxic (2.5 mg/L) stress for 96 h. We observed histological changes in the liver pre- and post-hypoxia and measured the changes in liver injury parameters. The results showed injury to the liver of L. polyactis following hypoxic stress, indicating that the liver is an important organ in the hypoxic stress response. To explore the genes and biological processes involved in hypoxic adaptation of L. polyactis at the overall transcription level, liver tissues were obtained pre- (0 h) and post- (6, 24, 48, and 96 h) hypoxia for transcriptomic analysis. GO and KEGG enrichment analysis of differentially expressed genes (DEGs) showed that DEGs were mainly enriched in redox equilibrium, metabolism, immunity, and apoptosis pathways under hypoxic stress. In addition, we examined the changes in molecular indices related to oxidative stress, cell apoptosis, and non-specific immunity in L. polyactis under hypoxic stress. The results showed that hypoxia induced reactive oxygen species overproduction, altered the activity of oxidase (MDA, PCO, 8-OhdG) and antioxidant enzymes (T-AOC, SOD, CAT, GPx, GR), and reduced the mRNA expression of key antioxidant genes (sod1, sod2, cat) in the liver, thus confirming that hypoxic stress induced oxidative stress. Additionally, we confirmed that hypoxic stress induced apoptosis in the liver by inducing the enzyme activity of Caspase-3/8/9 and the mRNA expression of key apoptotic genes (bcl2, bax, fas, and bnip3). Furthermore, hypoxic stress inhibited the activity of non-specific enzymes (AKP, ACP, and LZM) and mRNA expression of immune-related genes (tnf-α, il-1β, and il-17c); hence, hypoxic stress inhibited the non-specific immune function of L. polyactis. In conclusion, our study provides new insights into the molecular mechanisms underlying hypoxia adaptation in marine fish, which will aid in selective breeding of hypoxia-tolerant L. polyactis.
•Hypoxic stress induced liver tissue injury in small yellow croaker•Hypoxia induces DEGs related to oxidative stress, apoptosis, and immunity in the liver•Hypoxia altered enzyme activity and expression of genes related to antioxidants•Hypoxia altered Caspase-3/8/9 activity and expression of apoptosis genes•Hypoxia inhibited the activity of nonspecific immune-related enzymes and the expression of related genes•Hypoxia induces oxidative stress, apoptosis, and inhibits non-specific immunity
Cell lines play an important role in studying biological processes in vitro. In this study, we aimed to establish and characterize the first cell line isolated from the small yellow croaker ...(Larimichthys polyactis), an economically important marine fish in China. The small yellow croaker fry (SYCF) cells proliferated well in Leibovitz's 15 medium containing 2% antibiotics, 0.1 M NaCl, and 10% fetal bovine serum at 27 °C. Sequence analyses of mitochondrial genes confirmed the origin of SYCF cells. Furthermore, we unraveled the mechanism of hypoxia-induced apoptosis, a major threat to aquatic animals using the SYCF cell line. The HIF-PHD pathway of the SYCF cells can be activated under hypoxic stress, suggesting that this cell line is impressible for hypoxia and can serve as a superb model in vitro to study the cell physiology in L. polyactis in a hypoxic setting. We found that hypoxia results in morphological abnormalities, excess reactive oxygen species (ROS) production, and dysfunctional redox homeostasis, indicating the occurrence of oxidative stress in the SYCF cells. In addition, decreased cell viability and severe apoptosis were detected under hypoxic conditions. Through monitoring the indicators of the intrinsic and extrinsic apoptosis pathways, including the mitochondrial membrane potential, the expression of bax, bcl2, bnip3, and fas, Bcl2, and Caspase-3 protein, and activity of Caspases-3/8/9, we found that both pathways are involved. Considering that ROS-induced oxidative stress may be the main reason of apoptosis, we investigated whether the addition of an antioxidant, N-Acetyl-L-cysteine (NAC), will ameliorate the apoptosis. Interestingly, we found that both intrinsic and extrinsic apoptosis indicators were markedly recovered by adding NAC in culture media, significant remissions were also observed in the cell viability and apoptosis rate. These results suggested that hypoxia-induced apoptosis of SYCF cells may be mediated by the overproduction of ROS, and the antioxidant may be a useful material to relieve cell damage under hypoxia in fish. In general, this study established a marine fish cell line of SYCF cells, which can be used as an in vitro research model of L. polyactis and for revealing the mechanisms of ROS-induced apoptosis under hypoxic conditions in this species. This provides important insights into the cellular physiology of marine fish under hypoxia.
•The establishment and characterization of SYCF cells line•The HIF/PHD pathway in the SYCF cells could be activated under hypoxia•Hypoxia reduces oxidative stress and cell viability and causes severe intrinsic and extrinsic apoptosis in SYCF cells line•Hypoxia induced apoptosis of SYCF cells is mediated by ROS overproduction
Early weaning and shorter breastfeeding duration are applied by a proportion of young mothers, especially in the social spheres of poverty-stricken areas. Early childhood is a critical period for ...intestinal development, which is driven by intestinal stem cells (ISCs). However, how early weaning practice affects the function of ISCs to mediate intestinal development remains unclear.
We established an excellent early weaning mice model that has significant intestinal atrophy and growth arrest symptoms to explore the responses of ISCs to early weaning. The primary and passaged intestinal organoids from the suckling or early weaning mice were cultured to explore the underlying mechanism of early weaning affecting the ISCs.
Early weaning depressed the self-renewal of ISCs and attenuated the activity of ISCs-driven intestinal epithelial regeneration and crypt expansion in vivo and ex-vivo. Further results showed that early weaning retarded the differentiation of ISCs into transit-amplifying cells and Paneth cells, and accelerated the apoptosis of villous epithelial cells, jointly leading to intestinal epithelial atrophy. Mechanistically, early weaning inhibited Wnt signaling in ISCs, while an exogenous Wnt amplifier restored ISCs' function in ex-vivo.
Our findings indicate that early weaning depresses the activity of ISCs via attenuating Wnt/β-catenin signaling and triggers the proinflammatory cytokines TNF-α, IL-1β, IL-6, and IL-17 in jejunum, thereby impeding ISCs-driven epithelial regeneration and intestinal growth, which may provide a basal theory for the development of infant nutrients targeting stem cells to alleviate early weaning-induced intestinal problems.
Larimichthys crocea is an economically important marine fish in China. To date, the molecular mechanisms underlying testicular development and spermatogenesis in L. crocea have not been thoroughly ...elucidated. In this study, we conducted a comparative transcriptome analysis between testes (TES) and pooled multiple tissues (PMT) (liver, spleen, heart, and kidney) from six male individuals. More than 54 million clean reads were yielded from TES and PMT libraries. After mapping to the draft genome of L. crocea, we acquired 25,787 genes from the transcriptome dataset. Expression analyses identified a total of 3853 differentially expressed genes (DEGs), including 2194 testes-biased genes (highly expressed in the TES) and 1659 somatic-biased genes (highly expressed in the PMT). The dataset was further annotated by blasting with multi-databases. Functional genes and enrichment pathways involved in spermatogenesis and testicular development were analyzed, such as the neuroactive ligand-receptor interaction pathway, gonadotropin-releasing hormone (GnRH) and mitogen-activated protein kinase (MAPK) signaling pathways, cell cycle pathway, and dynein, kinesin, myosin, actin, heat shock protein (hsp), synaptonemal complex protein 2 (sycp2), doublesex- and mab-3-related transcription factor 1 (dmrt1), spermatogenesis-associated genes (spata), DEAD-Box Helicases (ddx), tudor domain-containing protein (tdrd), and piwi genes. The candidate genes identified by this study lay the foundation for further studies into the molecular mechanisms underlying testicular development and spermatogenesis in L. crocea.
KIF3A and KIF3B are homologous motor subunits of the Kinesin II protein family. KIF3A, KIF3B, and KAP3 form a heterotrimeric complex and play a significant role in spermatogenesis. Here, we first ...cloned full-length
kif3a
/
3b
cDNAs from
Larimichthys polyactis
.
Lp
-
kif3a/3b
are highly related to their homologs in other animals. The proteins are composed of three domains, an N-terminal head domain, a central stalk domain, and a C-terminus tail domain.
Lp
-
kif3a/3b
mRNAs were found to be ubiquitously expressed in the examined tissues, with high expression in the testis. Fluorescence in situ hybridization (FISH) was used to analyze the expression of
Lp
-
kif3a/3b
mRNAs during spermiogenesis. The results showed that
Lp
-
kif3a/3b
mRNAs had similar expression pattern and were continuously expressed during spermiogenesis. From middle spermatid to mature sperm,
Lp
-
kif3a/3b
mRNAs gradually localized to the side of the spermatid where the midpiece and tail form. In addition, we used immunofluorescence (IF) to observe that
Lp
-KIF3A protein co-localizes with tubulin during spermiogenesis. In early spermatid,
Lp
-KIF3A protein and microtubule signals were randomly distributed in the cytoplasm. In middle spermatid, however, the protein was detected primarily around the nucleus. In late spermatid, the protein migrated primarily to one side of the nucleus where the tail forms. In mature sperm,
Lp
-KIF3A and microtubules accumulated in the midpiece. Moreover,
Lp
-KIF3A co-localized with the mitochondria. In mature sperm,
Lp
-KIF3A and mitochondria were present in the midpiece. Therefore,
Lp-
KIF3A/KIF3B may be involved in spermiogenesis in
L. polyactis
, particularly during nuclear reshaping and tail formation.
The small yellow croaker
Larimichthys polyactis
is an economically important marine fish in Northeast Asia. Currently, its natural resources are threatened by overfishing and environmental pollution. ...Therefore, research on the reproductive system of the fish is crucial. Here, we studied the testis development and ultrastructural features of spermatogenesis in cultured
L. polyactis
using anatomical, histological, and ultrastructural techniques. A pair of testes, consisting of a central sperm duct and radial seminiferous lobules, were observed. The reproduction cycle of testes can be divided into stages I–VI. March to May was confirmed as the breeding season for male
L. polyactis
, while April is the ideal period for artificial breeding. The male
L. polyactis
can attain sexual maturity within 1 year. The spermatogenesis of
L. polyactis
comprised spermatogonium, spermatocyte, spermatid, and mature spermatozoon. The morphology of spermatogenic cells changed obviously during spermiogenesis, including nuclear shaping, midpiece and flagellum formation. The mature sperms consist of an ellipsoidal head, a short midpiece, and a long flagellum. The anterior of the head with a kidney-shaped nucleus can be distinguished. The midpiece is located posterior to the head and includes four to six spherical mitochondria. The flagellum has irregular lateral fins. The testis of
L. polyactis
is an unrestricted lobular type, with cystic spermatogenesis, type II spermiogenesis, and type II spermatozoa. These features are highly similar to those of other Sciaenid species. Our findings provide useful insights into the mechanism underlying testis development and spermatogenesis of
L. polyactis
, which can facilitate the artificial breeding of this species.
Phosphorus (P) pollution from modern swine production is a major environmental problem. Dietary interventions to promote bone growth can improve the utilization of dietary P, and thereby reduce its ...emission. Recent in vitro studies have shown that alpha-ketoglutarate (AKG) exerts a pro-osteogenic effect on osteoblast cells. This study aimed to evaluate the effects of AKG supplementation on bone growth, P and Ca digestion, and the gut microbial profile in piglets. Thirty-two piglets were randomly assigned into two dietary groups. The piglets were fed a basic diet containing 10 g/kg AKG or 10 g/kg maize starch (control) for 28 days. On days 21-28, titanium dioxide was used as an indicator to determine the apparent digestibility of P. AKG supplementation improved the bone mineral density, length, weight, and geometrical and strength properties of the femur and tibia. Furthermore, AKG supplementation increased apparent ileal and total tract digestibility of P. Colonic microbiota analysis results showed that AKG supplementation increased α-diversity and beneficial bacteria, including
and
, and decreased nitrogen fixation and chemoheterotrophy. Together, AKG supplementation improves bone growth, the utilization of dietary P, and the colonic microbial profile, which may provide a nutritional strategy for diminishing P pollution originating from the pig industry.
Piglets are particularly susceptible to oxidative stress, which causes inferior growth performance and intestinal damage. Squalene (SQ), a natural bioactive substance enriched in shark liver oil, ...shows excellent antioxidant properties and can currently be obtained at a low cost from deodorizer distillate during the production of plant oil. This study aimed to evaluate the effects of plant-derived SQ supplementation on the growth performance of piglets and explore the beneficial roles of SQ against oxidative stress and intestinal injury in diquat-challenged piglets. Forty piglets were randomly divided into five groups and fed a basal diet supplemented with SQ at 0, 500, 1000, or 2000 mg/kg for 5 weeks. Acute oxidative stress was induced in the piglets with diquat (10 mg/kg BW) at the fourth week of the experiment, followed by a 7-d recovery period. Results showed that before the diquat challenge, SQ supplementation significantly improved growth performance (average daily gain and feed conversion ratio) and serum antioxidant status, and after the diquat challenge, SQ supplementation significantly mitigated diquat-induced growth arrest, intestinal villous atrophy, intestinal epithelial cell apoptosis, intestinal hyperpermeability, and deficiency of intestinal epithelial tight junction proteins (zonula occludens-1, occludin, and claudin-3). Under oxidative stress induced by diquat, SQ supplementation consistently improved the antioxidant status of the small intestine, liver, and muscle. In vitro, SQ was shown to alleviate hydrogen peroxide (H2O2)-induced increase of the levels of intracellular reactive oxygen species and apoptosis of porcine intestinal epithelial cells. Taken together, SQ supplementation improves growth performance and effectively alleviates acute oxidative stress-induced growth retardation and intestinal injury via improving antioxidant capacity in piglets. Our findings may provide an efficient strategy for alleviating oxidative stress-induced inferior growth performance and intestinal damage in piglets.
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