OsUGE2 Regulates Plant Growth through Affecting ROS Homeostasis and Iron Level in Rice.

BACKGROUND: The growth and development of rice (Oryza sativa L.) are affected by multiple factors, such as ROS homeostasis and utilization of iron. Here, we demonstrate that OsUGE2, a gene encoding a UDP-glucose 4-epimerase, controls growth and development by regulating reactive oxygen species (ROS) and iron (Fe) level in rice. Knockout of this gene resulted in impaired growth, such as dwarf phenotype, weakened root growth and pale yellow leaves. Biochemical analysis showed that loss of function of OsUGE2 significantly altered the proportion and content of UDP-Glucose (UDP-Glc) and UDP-Galactose (UDP-Gal). Cellular observation indicates that the impaired growth may result from decreased cell length. More importantly, RNA-sequencing analysis showed that knockout of OsUGE2 significantly influenced the expression of genes related to oxidoreductase process and iron ion homeostasis. Consistently, the content of ROS and Fe are significantly decreased in OsUGE2 knockout mutant. Furthermore, knockout mutants of OsUGE2 are insensitive to both Fe deficiency and hydrogen peroxide (H2O2) treatment, which further confirmed that OsUGE2 control rice growth possibly through Fe and H2O2 signal. Collectively, these results reveal a new pathway that OsUGE2 could affect growth and development via influencing ROS homeostasis and Fe level in rice.

Cement-Augmented Screw Fixation with PHILOS Plating for Osteoporotic Proximal Humeral Fractures: An Observation of Mid- and Long-Term Curative Efficacy.

OBJECTIVES: The mid- and long-term clinical outcomes of cement-augmented screws in the treatment of osteoporotic proximal humeral fractures have rarely been reported. The aim of this study was to observe the mid- and long-term efficacy of combined cement-augmented screw fixation and PHILOS plating in the treatment of osteoporotic fractures of the proximal humerus. METHODS: This study retrospectively analyzed data from 19 patients with osteoporotic fractures of the proximal humerus who had undergone internal fixation at the Guizhou Provincial People's Hospital from February 2017 to May 2021. The cohort was comprised of six males and 13 females, aged 75-87 (mean age: 82.52 ± 1.24) years. According to the Neer classification, three, 12, and four patients had two-part, three-part, and four-part fractures, respectively. All patients were treated with open reduction internal fixation with cement-augmented screws and PHILOS plating. Time until fracture healing was recorded postoperatively. Patients were observed for postoperative complications, including humeral head necrosis, loosening or breaking of the augmented screws, screw perforation of the humeral head, and varus fracture displacement. Visual analog scale and Constant scores of the shoulder joint were compared 1, 3, 6, and 12 months after surgery. Scores at the most recent follow-up were used to evaluate shoulder joint function. Measured data conforming to a normal distribution were expressed as mean ± SD. Analysis of variance or rank sum tests were used for intergroup comparisons. A value of p < 0.05 was considered significant. RESULTS: All 19 patients followed up for 1-4 (average: 2.13 ± 0.61) years. Fractures united in all cases, with a healing time of 8-14 (average: 10.25 ± 1.72) weeks. There were no cases of humeral head necrosis, screw loosening, fractures, or perforation of the humeral head. One patient had mild varus fracture displacement with a reduced neck-shaft angle. There were significant differences in visual analog scale and Constant scores 1, 3, and 6 months after surgery (p < 0.05). The visual analog scale score was 0 at final follow-up in all cases. The Constant score of the shoulder joint was excellent, good, fair, and poor in two, 12, four, and one case, respectively, yielding an excellent and good rate of 73.68%. CONCLUSIONS: Cement-augmented screw fixation combined with PHILOS plating of osteoporotic proximal humeral fractures had good mid- and long-term clinical efficacy. It should be considered a new option for fracture treatment in such patients.

A metabolic intervention strategy to break evolutionary adaptability of tumor for reinforced immunotherapy.

The typical hallmark of tumor evolution is metabolic dysregulation. In addition to secreting immunoregulatory metabolites, tumor cells and various immune cells display different metabolic pathways and plasticity. Harnessing the metabolic differences to reduce the tumor and immunosuppressive cells while enhancing the activity of positive immunoregulatory cells is a promising strategy. We develop a nanoplatform (CLCeMOF) based on cerium metal-organic framework (CeMOF) by lactate oxidase (LOX) modification and glutaminase inhibitor (CB839) loading. The cascade catalytic reactions induced by CLCeMOF generate reactive oxygen species "storm" to elicit immune responses. Meanwhile, LOX-mediated metabolite lactate exhaustion relieves the immunosuppressive tumor microenvironment, preparing the ground for intracellular regulation. Most noticeably, the immunometabolic checkpoint blockade therapy, as a result of glutamine antagonism, is exploited for overall cell mobilization. It is found that CLCeMOF inhibited glutamine metabolism-dependent cells (tumor cells, immunosuppressive cells, etc.), increased infiltration of dendritic cells, and especially reprogrammed CD8+ T lymphocytes with considerable metabolic flexibility toward a highly activated, long-lived, and memory-like phenotype. Such an idea intervenes both metabolite (lactate) and cellular metabolic pathway, which essentially alters overall cell fates toward the desired situation. Collectively, the metabolic intervention strategy is bound to break the evolutionary adaptability of tumors for reinforced immunotherapy.

Ethylene accelerates copper oxide nanoparticle-induced toxicity at physiological, biochemical, and ultrastructural levels in rice seedlings.

The enormous use of metal-based nanoparticles (NPs) in different sectors may result in enhanced accumulation in agricultural soil, which could impose negative effects on crop productivity. Hence, strategies are needed to explore the mechanisms of copper oxide nanoparticle (CuO NP)-induced toxicity in crops. The present study aimed to investigate the involvement of ethylene in CuO NP-induced toxicity in rice seedlings. Here, our results indicate that 450 mg L-1 of CuO NPs induced toxic effects in rice seedlings. Thus, it was evidenced by the reduced plant biomass accumulation, enhanced oxidative stress indicators, and cellular ultrastructural damages. More importantly, the exogenous supply of ethylene biosynthesis and signaling antagonists cobalt (Co) and silver (Ag) respectively provided tolerance and improved the defense system of rice seedlings against CuO NP toxicity. The ethylene antagonists could significantly reduce the extent of ultrastructural and stomatal damage by controlling the ROS accumulation in rice seedlings under CuO NP stress. Furthermore, Co and Ag augmented the antioxidant defense system against CuO NP-induced toxicity. Contrary to that, all oxidative damage attributes were further enhanced exogenous application of ethylene biosynthesis precursor [1-aminocyclopropane-1-carboxylic acid (ACC)] in the presence of CuO NPs. In addition, ACC could increase the CuO NP-induced stomatal and ultrastructural damages by reducing the ROS-scavenging ability in rice seedlings. Taken together, these results indicate the involvement of ethylene in CuO NP-induced toxicity in rice seedlings.

Cd248a and Cd248b in zebrafish participate in innate immune responses.

CD248, also known as endosialin or tumor endothelial marker 1, is a type I single transmembrane glycoprotein. CD248 has been demonstrated to be upregulated in cancers, tumors and many fibrotic diseases in human and mice, such as liver damage, pulmonary fibrosis, renal fibrosis, arthritis and tumor neovascularization. However, no definite CD248 orthologs in fish have been documented so far. In this study, we report the identification of cd248a and cd248b in the zebrafish. Both the phylogenetic analysis and the conserved synteny strongly suggested that zebrafish cd248a and cd248b are orthologs of the human CD248. Both cd248a and cd248b exhibited similar and dynamic expression pattern in early development, both genes had weak maternal expression, the zygotic transcripts were first seen in anterior somites and head mesenchyme, then shifted to eyes and head mesenchyme, later expanded to branchial arches, and gradually declined with development. The expression profiles of cd248a and cd248b were upregulated upon LPS (Lipopolysaccharide) challenge. Both Cd248a protein and Cd248b protein were localized on the cell membrane and cytoplasm, and overexpression of cd248a and cd248b induced the expression of pro-inflammatory cytokines, in vitro and in vivo. Moreover, deficiency of cd248a or cd248b both downregulated the expression of pro-inflammatory cytokines and upregulated anti-inflammatory cytokine. Additionally, loss of cd248a or cd248b both downregulated the expression of pro-inflammatory cytokines after LPS treatment. Taken together, these results indicated that cd248a and cd248b in zebrafish were involved in immune response and would provide further information to understand functions of Cd248 protein in innate immunity of fish.

Ethylene participates in zinc oxide nanoparticles induced biochemical, molecular and ultrastructural changes in rice seedlings.

Nowadays, the applications of engineered nanoparticles (ENPs) have been significantly increased, thereby negatively affecting crop production and ultimately contaminating the food chain worldwide. Zinc oxide nanoparticles (ZnO NPs) induced oxidative stress has been clarified in previous studies. But until now, it has not been investigated that how ethylene mediates or participates in ZnO NPs-induced toxicity and related cellular ultrastructural changes in rice seedlings. Here, we reported that 500 mg/L of ZnO NPs reduced the fresh weight (54.75% and 55.64%) and dry weight (40.33% and 47.83%) in shoot and root respectively as compared to control. Furthermore, ZnO NPs (500 mg/L) reduced chlorophyll content (72% Chla, 70% Chlb), induced the stomatal closure and ultrastructural damages by causing oxidative stress in rice seedlings. These cellular damages were significantly increased by exogenous applications of ethylene biosynthesis precursor (ACC) in the presence of ZnO NPs. In contrary, ZnO NPs induced damages on the above-mentioned attributes were reversed through the exogenous supply of ethylene signaling and biosynthesis antagonists such as silver (Ag) and cobalt (Co) respectively. Interestingly, ZnO NPs accelerate ethylene biosynthesis by up-regulating the transcriptome of ethylene biosynthesis responsive genes. The antioxidant enzymes activities and related gene expressions were further increased in ethylene signaling and biosynthesis associated antagonists (Ag and Co) treated seedlings as compared to sole ZnO NPs treatments. In contrary, the above-reported attributes were further decreased by ACC together with ZnO NPs. In a nutshell, ethylene effectively contributes in ZnO NPs induced toxicity and causing ultrastructural and stomatal damage in rice seedlings. Such findings could have potential implications in producing genetic engineered crops, which will be able to tolerate nanoparticles toxicity in the environment.

Conservation of eATP perception throughout multicellular animal evolution: Identification and functional characterization of coral and amphioxus P2X7-like receptors and flounder P2X7 receptor.

Perception of extracellular ATP (eATP), a common endogenous damage-associated molecular pattern, is through its receptor P2X7R. If eATP/P2X7R signaling is conserved throughout animal evolution is unknown. Moreover, little information is currently available regarding P2X7R in invertebrates. Here we demonstrated that the coral P2X7-like receptor, AdP2X7RL, the amphioxus P2X7-like receptor, BjP2X7RL and the flounder P2X7 receptor, PoP2X7R, shared common features characteristic of mammalian P2X7R, and their 3D structures displayed high resemblance to that of human P2X7R. Expression of Adp2x7rl, Bjp2x7rl and Pop2x7r was all subjected to the regulation by LPS and ATP. We also showed that AdP2X7RL, BjP2X7RL and PoP2X7R were distributed on the plasma membrane in AdP2X7RL-, BjP2X7RL- and PoP2X7R-expressing HEK cells, and had strong affinity to eATP. Importantly, the binding of AdP2X7RL, BjP2X7RL and PoP2X7R to eATP all induced similar downstream responses, including induction of cytokines (IL-1ß, IL-6, IL-8 and CCL-2), enhancement of phagocytosis and activation of AKT/ERK-associated signaling pathway observed for mammalian P2X7R. Collectively, our results indicate for the first time that both coral and amphioxus P2X7RL as well as flounder P2X7R can interact with eATP, and induce events that trigger mammalian mechanisms, suggesting the high conservation of eATP perception throughout multicellular animal evolution.