Naringenin mitigates cadmium-induced cell death, oxidative stress, mitochondrial dysfunction, and inflammation in KGN cells by regulating the expression of sirtuin-1.

The objective of this study was to examine the potential protective role of naringenin against the harmful effects induced by cadmium in KGN cell line. Cell viability was evaluated by cell counting kit-8 assay. Caspase-3/-9 activities were determined by caspase-3/-9 activity assay kits, respectively. Intracellular reactive oxygen species (ROS) level was detected by ROS-Glo™ H2O2 Assay, antioxidant capacity was determined by a total antioxidant capacity assay kit. Mitochondrial membrane potential (MMP), ATP level, and ATP synthase activity were determined by JC-1, ATP assay kit, and ATP synthase activity assay kit, respectively. The mRNA expression was determined by qRT-PCR. Cadmium reduced cell viability and increased caspase-3/-9 activities in a concentration-dependent manner. Naringenin improved cell viability and reduced caspase-3/-9 activities in cadmium-stimulated KGN cells in a concentration-dependent manner. Cadmium diminished the antioxidant capacity, increased ROS production, and induced mitochondrial dysfunction in KGN cells. These effects were ameliorated by naringenin treatment in a concentration-dependent manner. Furthermore, naringenin reduced the levels of pro-inflammatory cytokines in KGN cells exposed to cadmium. SIRT1 knockdown downregulated its expression in KGN cells and compromised the protective effects of naringenin on cell viability and caspase-3/-9 activities in cadmium-stimulated KGN cells. Naringenin prevented the reduction of MMP, ATP levels, and ATP synthase activity in cadmium-stimulated KGN cells in a concentration-dependent manner. However, these protective effects were significantly reversed by SIRT1 knockdown. In conclusion, this study suggests that naringenin protects against cadmium-induced damage by regulating oxidative stress, mitochondrial function, and inflammation in KGN cells, with SIRT1 playing a potential mediating role.

Selective autophagic degradation of ACLY (ATP citrate lyase) maintains citrate homeostasis and promotes oocyte maturation.

Macroautophagy/autophagy is a cellular and energy homeostatic mechanism that contributes to maintain the number of primordial follicles, germ cell survival, and anti-ovarian aging. However, it remains unknown whether autophagy in granulosa cells affects oocyte maturation. Here, we show a clear tendency of reduced autophagy level in human granulosa cells from women of advanced maternal age, implying a potential negative correlation between autophagy levels and oocyte quality. We therefore established a co-culture system and show that either pharmacological inhibition or genetic ablation of autophagy in granulosa cells negatively affect oocyte quality and fertilization ability. Moreover, our metabolomics analysis indicates that the adverse impact of autophagy impairment on oocyte quality is mediated by downregulated citrate levels, while exogenous supplementation of citrate can significantly restore the oocyte maturation. Mechanistically, we found that ACLY (ATP citrate lyase), which is a crucial enzyme catalyzing the cleavage of citrate, was preferentially associated with K63-linked ubiquitin chains and recognized by the autophagy receptor protein SQSTM1/p62 for selective autophagic degradation. In human follicles, the autophagy level in granulosa cells was downregulated with maternal aging, accompanied by decreased citrate in the follicular fluid, implying a potential correlation between citrate metabolism and oocyte quality. We also show that elevated citrate levels in porcine follicular fluid promote oocyte maturation. Collectively, our data reveal that autophagy in granulosa cells is a beneficial mechanism to maintain a certain degree of citrate by selectively targeting ACLY during oocyte maturation.Abbreviations: 3-MA: 3-methyladenine; ACLY: ATP citrate lyase; AMA: advanced maternal age; CG: cortical granule; CHX: cycloheximide; CQ: chloroquine; CS: citrate synthase; COCs: cumulus-oocyte-complexes; GCM: granulosa cell monolayer; GV: germinal vesicle; MII: metaphase II stage of meiosis; PB1: first polar body; ROS: reactive oxygen species; shRNA: small hairpin RNA; SQSTM1/p62: sequestosome 1; TCA: tricarboxylic acid; TOMM20/TOM20: translocase of outer mitochondrial membrane 20; UBA: ubiquitin-associated domain; Ub: ubiquitin; WT: wild-type.

Substitution of manure for mineral P fertilizers increases P availability by enhancing microbial potential for organic P mineralization in greenhouse soil.

The shortage of phosphorus (P) as a resource represents a major challenge for the sustainable development of agriculture. Manure has a high P content and is a potential substitute for mineral P fertilizers. However, little is known about the effects on soil P availability and soil microbial P transformation of substituting manure for mineral P fertilizers. In this study, variations in soil P availability and bacterial P mobilization were evaluated under treatment with manure as compared to mineral P fertilizers. In the greenhouse fruit and vegetable production system that provided the setting for the study, substitution of manure for mineral P (PoR treatment) resulted in a similar level of soil total P and a similar fruit and vegetable yield as compared to traditional fertilization, but a significantly increased level of soil available P. In addition, PoR treatment enhanced bacterial organic P mineralization potential and decreased inorganic P dissolution potential. These results demonstrate that manure application increases the availability of soil P primarily by enhancing soil microbial Po mineralization, indicating the potential feasibility of applying manure instead of mineral P fertilizers in greenhouse farming.

Effects of intrauterine infusion of platelet-rich plasma on hormone levels and endometrial receptivity in patients with repeated embryo implantation failure.

OBJECTIVE: To analyze the effect of intrauterine infusion on platelet-rich plasma on hormone levels and endometrial receptivity of patients with repeated embryo implantation failure (RIF). METHODS: A total of 64 patients with repeated implantation failure and re-fertilization-embryo transfer who were admitted to our hospital from January 2019 to December 2021 were analyzed retrospectively. Among them were 30 patients who did not receive the platelet-rich plasma perfusion therapy. This became the control group (CG). The 34 patients who received the therapy were regarded as the research group (RG). The changes of hormone levels before and after the treatment and endometrial receptivity after the treatment were evaluated. The outcomes of IVF assisted pregnancy, including rates of embryo implantation, clinical pregnancy, and early miscarriage, were compared after the treatment. Risk factors for clinical pregnancy were analyzed by logistic regression. RESULTS: After treatment, the estradiol (E2) level increased and the follicle stimulating hormone (FSH) level decreased (P<0.05), but there was no marked difference in luteinizing hormone (LH) before or after the treatment (P>0.05). The E2 level in the RG was higher than that in the CG, and FSH in the RG was lower (P<0.05). In comparison to CG, the endometrial thickness on the day of human chorionic gonadotropin (hCG) injection and embryo transfer in the RG increased dramatically (P<0.05). The uterine artery pulsation index (PI) and uterine artery resistance index (RI) decreased (P<0.05). The embryo implantation and clinical pregnancy rates in the RG increased markedly (P<0.01), and the early abortion rate decreased significantly (P<0.05). The logistic regression analysis identified that age, number of transplant failures, treatment regimens, and FSH were risk factors for clinical pregnancy outcomes in patients. CONCLUSION: Intrauterine infusion of platelet-rich plasma can improve the hormone levels in RIF patients, increase endometrial thickness, and enhance endometrial blood flow, increasing the pregnancy rate of patients and improving clinical pregnancy.

Changes in phosphorus mobilization and community assembly of bacterial and fungal communities in rice rhizosphere under phosphate deficiency.

Rhizosphere microorganisms are closely associated with phosphorus (P) uptake in plants and are considered potential agents to mitigate P shortage. However, the mechanisms of rhizospheric microbial community assembly under P deficiency have yet to be elucidated. In this study, bacterial and fungal communities in rice rhizosphere and their P mobilization potential under high (+P) and low (-P) concentrations of P were investigated. Bacterial and fungal community structures were significantly different between -P and +P treatments. And both bacterial and fungal P-mobilizing taxa were enriched in-P treatment; however, the proportion of P-mobilizing agents in the fungal community was markedly greater than that in the bacterial community. A culture experiment confirmed that microbial phosphate solubilizing capacity was significantly higher in -P treatment compared with that in +P treatment. -P treatment lowered bacterial diversity in rice rhizosphere but increased fungal diversity. Further analysis demonstrated that the contribution of deterministic processes in governing bacterial community assembly was strengthened under P deficiency but was largely weakened in shaping the fungal community. These results highlighted that enriching P-mobilizing microbes in the rhizosphere is a vital way for rice to cope with P deficiency, and that fungi contribute considerably to P mobilization in rice rhizosphere. Findings from the study provide novel insights into the assembly of the rhizosphere microbiome under P deficiency and this will facilitate the development of rhizosphere microbial regulation strategies to increase nutrient uptake in plants.

Resveratrol regulates the inflammation and oxidative stress of granulosa cells in PCOS via targeting TLR2.

Polycystic ovary syndrome (PCOS) is featured as a common endocrine disorder in reproductive-aged women, while its pathophysiology is not fully illustrated. This study examined potential actions of resveratrol in PCOS cellular model and explored the underlying interaction between resveratrol and toll-like receptor 2 (TLR2). This study performed the bioinformatics analysis on two microarray datasets (GSE34526 and GSE138518). We found that TLR2 was one of potential hub genes that may be associated with PCOS. Further examination showed that TLR2 was highly expressed in granulosa cells from PCOS group compared with control. The in vitro studies showed that LPS intervention caused an increased expression of TLR2 and the pro-inflammatory mediators, and induced oxidative stress in the granulosa cells, which was concentration-dependently antagonized by resveratrol treatment. TLR2 silence significantly attenuated LPS-induced increase TNF-α, IL-1ß, IL-6 and IL-8 expression and oxidative stress of granulosa cells. Furthermore, TLR2 overexpression promoted inflammatory response and oxidative stress in the granulosa cells, which was antagonized by resveratrol treatment. In conclusion, resveratrol could attenuate LPS-induced inflammation and oxidative stress in granulosa cells, and the underlying mechanisms may be related to the inhibitory effect of resveratrol on TLR2 expression in granulosa cells.

Camellia sinensis Chloroplast Fluoride Efflux Gene CsABCB9 Is Involved in the Fluoride Tolerance Mechanism.

Soil is a main source of fluoride for plants. The tea plants (Camellia sinensis) accumulate excessive amounts of fluoride in their leaves compared to other plants, but their fluoride tolerance mechanism is poorly understood. A chloroplast fluoride efflux gene (CsABCB9) was newly discovered by using transcriptome analysis, cloned from Camellia sinensis, and its function was demonstrated in the fluoride detoxication mechanism in Escherichia coli/Xenopus laevis oocytes and Arabidopsis thaliana. CsABCB9 is expressed in tea leaves upon F− treatment. The growth of tea, E. coli, and Arabidopsis were inhibited by F− treatment. However, growth of CsABCB9-overexpression in E. coli was shown to increase with lower fluoride content under F− treatment compared to the control. Furthermore, chlorophyll, xanthophyll and soluble sugar contents of CsABCB9-overexpression in Arabidopsis were improved under F− treatment compared to the wild type. CsABCB9 functions in fluoride transport, and the mechanism by which CsABCB9 improves fluoride resistance in tea is mainly chloroplast protection through fluoride efflux.

The enrichment of sugars and phenols from fast pyrolysis of bamboo via ethanol-Fenton pretreatment.

The high-purity compounds (e.g., sugars and phenols) are important raw materials and chemicals, which can be produced by biomass pyrolysis. However, the direct biomass pyrolysis produces complex compounds and thus inhibiting its large-scale utilization. To increase the yield and enrichment of sugars and phenols, a green coupling process based on ethanol-Fenton pretreatment combined with fast pyrolysis is firstly proposed. The bamboo was effectively separated into the ethanol-Fenton pretreated bamboo (EF-bamboo), lignin-rich fractions, and hemicellulose-degradation intermixtures with the massive removal of inorganic metals via this process. Compared with the fast pyrolysis of raw bamboo, the levoglucosan yield of EF-bamboo increased 5.4 times and the enrichment of sugars improved from 7.6% to 59.7%. Similarly, the yield of monophenols from lignin-rich fractions increased around 0.6 times and the enrichment of monophenols increased from 25.7% to 63.5%. This work provides a green and efficient route to produce high-yield and high-enrichment sugars and phenols.

Mapping combined with principal component analysis identifies excellent lines with increased rice quality.

Quality-related traits are some of the most important traits in rice, and screening and breeding rice lines with excellent quality are common ways for breeders to improve the quality of rice. In this study, we used 151 recombinant inbred lines (RILs) obtained by crossing the northern cultivated japonica rice variety ShenNong265 (SN265) with the southern indica rice variety LuHui99 (LH99) and simplified 18 common rice quality-related traits into 8 independent principal components (PCs) by principal component analysis (PCA). These PCs included peak and hot paste viscosity, chalky grain percentage and chalkiness degree, brown and milled rice recovery, width length rate, cooked taste score, head rice recovery, milled rice width, and cooked comprehensive score factors. Based on the weight ratio of each PC score, the RILs were classified into five types from excellent to poor, and five excellent lines were identified. Compared with SN265, these 5 lines showed better performance regarding the chalky grain percentage and chalkiness degree factor. Moreover, we performed QTL localization on the RIL population and identified 94 QTLs for quality-related traits that formed 6 QTL clusters. In future research, by combining these QTL mapping results, we will be using backcrossing to aggregate excellent traits and achieve quality improvement of SN265.

An efficient way to synthesize biomass-based molybdenum carbide catalyst via pyrolysis carbonization and its application for lignin catalytic pyrolysis.

In this study, a simple and rapid method was proposed to synthesize orthorhombic α-Mo2C as catalyst for lignin catalytic pyrolysis. Biomass in-situ pyrolysis products were used as the carbon source and supporter, the carbonization of Mo precursor was realized under rapid heating. Experimental results show that Pine-Mo2C catalyst can achieve lignin pyrolysis vapor bond breaking and deoxidation under normal pressure, and the yield of monocyclic aromatic hydrocarbons is 13.26 wt%, of which aromatic hydrocarbons with side chain account for 74%. The side chain aliphatic hydrocarbons of lignin are effectively retained, and hydrogen consumption is minimized. The characterization of catalyst and experiments of guaiacol, 2-phenoxy-1-phenylethanol and 4,4'-biphenol shows that efficient deoxidation is due to targeted attack of catalyst on C-O. Therefore, Pine-Mo2C shows excellent activity in promoting direct bond breaking deoxidation of lignin.

Defective chromatin architectures in embryonic stem cells derived from somatic cell nuclear transfer impair their differentiation potentials.

Nuclear transfer embryonic stem cells (ntESCs) hold enormous promise for individual-specific regenerative medicine. However, the chromatin states of ntESCs remain poorly characterized. In this study, we employed ATAC-seq and Hi-C techniques to explore the chromatin accessibility and three-dimensional (3D) genome organization of ntESCs. The results show that the chromatin accessibility and genome structures of somatic cells are re-arranged to ESC-like states overall in ntESCs, including compartments, topologically associating domains (TADs) and chromatin loops. However, compared to fertilized ESCs (fESCs), ntESCs show some abnormal openness and structures that have not been reprogrammed completely, which impair the differentiation potential of ntESCs. The histone modification H3K9me3 may be involved in abnormal structures in ntESCs, including incorrect compartment switches and incomplete TAD rebuilding. Moreover, ntESCs and iPSCs show high similarity in 3D genome structures, while a few differences are detected due to different somatic cell origins and reprogramming mechanisms. Through systematic analyses, our study provides a global view of chromatin accessibility and 3D genome organization in ntESCs, which can further facilitate the understanding of the similarities and differences between ntESCs and fESCs.

Nano-Mechanical Properties and Creep Behavior of Ti6Al4V Fabricated by Powder Bed Fusion Electron Beam Additive Manufacturing.

Effects of scanning strategy during powder bed fusion electron beam additive manufacturing (PBF-EB AM) on microstructure, nano-mechanical properties, and creep behavior of Ti6Al4V alloys were compared. Results show that PBF-EB AM Ti6Al4V alloy with linear scanning without rotation strategy was composed of 96.9% α-Ti and 2.7% ß-Ti, and has a nanoindentation range of 4.11-6.31 GPa with the strain rate ranging from 0.001 to 1 s-1, and possesses a strain-rate sensitivity exponent of 0.053 ± 0.014. While PBF-EB AM Ti6Al4V alloy with linear and 90° rotate scanning strategy was composed of 98.1% α-Ti and 1.9% ß-Ti and has a nanoindentation range of 3.98-5.52 GPa with the strain rate ranging from 0.001 to 1 s-1, and possesses a strain-rate sensitivity exponent of 0.047 ± 0.009. The nanohardness increased with increasing strain rate, and creep displacement increased with the increasing maximum holding loads. The creep behavior was mainly dominated by dislocation motion during deformation induced by the indenter. The PBF-EB AM Ti6Al4V alloy with only the linear scanning strategy has a higher nanohardness and better creep resistance properties than the alloy with linear scanning and 90° rotation strategy. These results could contribute to understanding the creep behavior of Ti6Al4V alloy and are significant for PBF-EB AM of Ti6Al4V and other alloys.

Overexpression of the High-Affinity Nitrate Transporter OsNRT2.3b Driven by Different Promoters in Barley Improves Yield and Nutrient Uptake Balance.

Improving nitrogen use efficiency (NUE) is very important for crops throughout the world. Rice mainly utilizes ammonium as an N source, but it also has four NRT2 genes involved in nitrate transport. The OsNRT2.3b transporter is important for maintaining cellular pH under mixed N supplies. Overexpression of this transporter driven by a ubiquitin promoter in rice greatly improved yield and NUE. This strategy for improving the NUE of crops may also be important for other cereals such as wheat and barley, which also face the challenges of nutrient uptake balance. To test this idea, we constructed transgenic barley lines overexpressing OsNRT2.3b. These transgenic barley lines overexpressing the rice transporter exhibited improved growth, yield, and NUE. We demonstrated that NRT2 family members and the partner protein HvNAR2.3 were also up-regulated by nitrate treatment (0.2 mM) in the transgenic lines. This suggests that the expression of OsNRT2.3b and other HvNRT2 family members were all up-regulated in the transgenic barley to increase the efficiency of N uptake and usage. We also compared the ubiquitin (Ubi) and a phloem-specific (RSs1) promoter-driven expression of OsNRT2.3b. The Ubi promoter failed to improve nutrient uptake balance, whereas the RSs1 promoter succeed in increasing the N, P, and Fe uptake balance. The nutrient uptake enhancement did not include Mn and Mg. Surprisingly, we found that the choice of promoter influenced the barley phenotype, not only increasing NUE and grain yield, but also improving nutrient uptake balance.

Overexpression of Nitrate Transporter OsNRT2.1 Enhances Nitrate-Dependent Root Elongation.

Root morphology is essential for plant survival. NO3- is not only a nutrient, but also a signal substance affecting root growth in plants. However, the mechanism of NO3--mediated root growth in rice remains unclear. In this study, we investigated the effect of OsNRT2.1 on root elongation and nitrate signaling-mediated auxin transport using OsNRT2.1 overexpression lines. We observed that the overexpression of OsNRT2.1 increased the total root length in rice, including the seminal root length, total adventitious root length, and total lateral root length in seminal roots and adventitious roots under 0.5-mM NO3⁻ conditions, but not under 0.5-mM NH4⁺ conditions. Compared with wild type (WT), the 15NO3- influx rate of OsNRT2.1 transgenic lines increased by 24.3%, and the expressions of auxin transporter genes (OsPIN1a/b/c and OsPIN2) also increased significantly under 0.5-mM NO3- conditions. There were no significant differences in root length, ß-glucuronidase (GUS) activity, and the expressions of OsPIN1a/b/c and OsPIN2 in the pDR5::GUS transgenic line between 0.5-mM NO3- and 0.5-mM NH4⁺ treatments together with N-1-naphthylphalamic acid (NPA) treatment. When exogenous NPA was added to 0.5-mM NO3- nutrient solution, there were no significant differences in the total root length and expressions of OsPIN1a/b/c and OsPIN2 between transgenic plants and WT, although the 15NO3- influx rate of OsNRT2.1 transgenic lines increased by 25.2%. These results indicated that OsNRT2.1 is involved in the pathway of nitrate-dependent root elongation by regulating auxin transport to roots; i.e., overexpressing OsNRT2.1 promotes an effect on root growth upon NO3- treatment that requires active polar auxin transport.

Overexpression of a High-Affinity Nitrate Transporter OsNRT2.1 Increases Yield and Manganese Accumulation in Rice Under Alternating Wet and Dry Condition.

Nitrate and manganese (Mn) are necessary elements for the growth and development of rice in paddy soil. Under physiological conditions, we previously reported that the uptake of Mn in roots can be improved by the addition of external nitrate but not ammonium. To investigate the mechanism(s) of this phenotype, we produced plant lines overexpressing OsNRT2.1 and assessed Mn uptake under alternating wet and dry (AWD) and waterlogged (WL) conditions. Under AWD condition, we observed a 31% reduction in grain yields of wild type (WT) plants compared to WL condition. Interestingly, the overexpression of OsNRT2.1 could recover this loss, as OsNRT2.1 transgenic lines displayed higher grain yields than WT plants. We also observed 60% higher grain Mn in the transgenic lines in AWD condition and approximately 30% higher Mn in the grain of transgenic lines in WL condition. We further found that the overexpression of OsNRT2.1 did not alter Mg and Fe in the seeds in either growth condition. The reasons for the increased Mn content in OsNRT2.1 transgenic seeds in AWD condition could be explained by the elevated expression of OsNRAMP family genes including OsNRAMP3, OsNRAMP5, and OsNRAMP6 in node I, the panicle-neck, and the flag leaves. The mechanism(s) underpinning the upregulation of these genes requires further investigation. Taken together, our results provide a new function of OsNRT2.1 in improving rice yields and grain Mn accumulation during water-saving cultivation patterns. This represents a new strategy for maintaining yield and improving food quality in a sustainable agricultural system.

OsCHX14 is Involved in the K+ Homeostasis in Rice (Oryza sativa) Flowers.

Previously we showed in the osjar1 mutants that the lodicule senescence which controls the closing of rice flowers was delayed. This resulted in florets staying open longer when compared with the wild type. The gene OsJAR1 is silenced in osjar1 mutants and is a key member of the jasmonic acid (JA) signaling pathway. We found that K concentrations in lodicules and flowers of osjar1-2 were significantly elevated compared with the wild type, indicating that K+ homeostasis may play a role in regulating the closure of rice flowers. The cation/H+ exchanger (CHX) family from rice was screened for potential K+ transporters involved as many members of this family in Arabidopsis were exclusively or preferentially expressed in flowers. Expression profiling confirmed that among 17 CHX genes in rice, OsCHX14 was the only member that showed an expression polymorphism, not only in osjar1 mutants but also in RNAi (RNA interference) lines of OsCOI1, another key member of the JA signaling pathway. This suggests that the expression of OsCHX14 is regulated by the JA signaling pathway. Green fluorescent protein (GFP)-tagged OsCHX14 protein was preferentially localized to the endoplasmic reticulum. Promoter-ß-glucuronidase (GUS) analysis of transgenic rice revealed that OsCHX14 is mainly expressed in lodicules and the region close by throughout the flowering process. Characterization in yeast and Xenopus laevis oocytes verified that OsCHX14 is able to transport K+, Rb+ and Cs+ in vivo. Our data suggest that OsCHX14 may play an important role in K+ homeostasis during flowering in rice.

Overexpression of a pH-sensitive nitrate transporter in rice increases crop yields.

Cellular pH homeostasis is fundamental for life, and all cells adapt to maintain this balance. In plants, the chemical form of nitrogen supply, nitrate and ammonium, is one of the cellular pH dominators. We report that the rice nitrate transporter OsNRT2.3 is transcribed into two spliced isoforms with a natural variation in expression ratio. One splice form, OsNRT2.3b is located on the plasma membrane, is expressed mainly in the phloem, and has a regulatory motif on the cytosolic side that acts to switch nitrate transport activity on or off by a pH-sensing mechanism. High OsNRT2.3b expression in rice enhances the pH-buffering capacity of the plant, increasing N, Fe, and P uptake. In field trials, increased expression of OsNRT2.3b improved grain yield and nitrogen use efficiency (NUE) by 40%. These results indicate that pH sensing by the rice nitrate transporter OsNRT2.3b is important for plant adaption to varied N supply forms and can provide a target for improving NUE.