The discovery of a novel single-function intermolecular Diels-Alder enzyme for the biosynthesis of hetero-dimer lithocarpins.

The deep-sea fungus Phomopsis lithocarpus FS508 produces tenellone-macrolide conjugated hetero-dimer lithocarpins A-G with anti-tumor activities. The deficiency of new intermolecular Diels-Alder (DA) enzymes hindered the development of new bioactive hetero-dimers. A novel single-function intermolecular DA enzyme, g7882, was initially discovered in this study. The deletion of g7882 led to the disappearance of lithocarpin A and an increase in precursor level . the overexpression of g7882 significantly improved lithocarpin A yield. The in vitro function of g7882DA was also confirmed by biochemical reaction using tenellone B as a substrate. Additionally, the knockout of KS modules of PKS in cluster 41 and cluster 81 (lit cluster) eliminated the production of lithocarpins, which firstly explains the biosynthetic process of hetero-dimer lithocarpins mediated by DA enzyme in FS508. Furthermore, the removal of a novel acetyltransferase GPAT in cluster 41 and the oxidoreductase, prenyltransferase in cluster81 resulted in the reduction of lithocarpin A in P. lithocarpus. The overexpression of gpat in P. lithocarpus FS508 improved the yield of lithocarpin A significantly and produced a new tenellone derivative lithocarol G. This study offers a new DA enzyme tool for the biosynthesis of novel hetero-dimer and biochemical clues for the biosynthetic logic elucidation of lithocarpins.

Exogenous Spermidine and Amino-Ethoxyvinylglycine Improve Nutritional Quality via Increasing Amino Acids in Rice Grains.

Polyamines and ethylene are key regulators of the growth and development, quality formation, and stress response of cereal crops such as rice. However, it remains unclear whether the application of these regulators could improve the nutritional quality via increasing amino acids in rice grains. This study examined the role of exogenous polyamines and ethylene in regulating amino acid levels in the milled rice of earlier-flowered superior grain (SG) and later-flowered inferior grain (IG). Two rice varieties were field grown, and either 1 mmol L-1 spermidine (Spd) or 50 µmol L-1 amino-ethoxyvinylglycine (AVG) was applied to panicles at the early grain-filling stage. The control check (CK) was applied with deionized water. The results showed that the Spd or AVG applications significantly increased polyamine (spermine (Spm) and Spd) contents and decreased ethylene levels in both SG and IG and significantly increased amino acid levels in the milled rice of SG and IG relative to the CK. Collectively, the application of Spd or AVG can increase amino acid-based nutritional quality and grain yield via increasing polyamine (Spm and Spd) contents and reducing ethylene levels in both SG and IG of rice.

3-Carboxy-indole derivatives from the deep-sea-derived fungus Phomopsis tersa FS441.

Three new compounds phomtersines A-C (1-3) together with nine known compounds were isolated from the marine-derived fungus Phomopsis tersa FS441. Their structures were sufficiently established by spectroscopic methods, including extensive 1D and 2D NMR techniques and modified Snatzke's method. Moreover, compounds 1-12 were evaluated for cytotoxic and anti-inflammatory activities. As a result, phomtersine B (2) and the known compound 10 showed moderate cytotoxic activity against the four tested cell lines with IC50 values ranging from 20.21 to 36.53 µM, and phomtersine A (1) exhibited moderate inhibitory activity against LPS-induced NO production.

Involvement of brassinosteroids and abscisic acid in spikelet degeneration in rice under soil drying during meiosis.

Spikelet degeneration in rice (Oryza sativa L.) is a serious physiological defect, and can be regulated by soil moisture status and phytohormones. This study investigated the possibility that brassinosteroids (BRs) in collaboration with abscisic acid (ABA) are involved in mediating the effect of soil drying during meiosis on spikelet degeneration in rice. Three rice cultivars were field grown and three irrigation regimes including well watered (WW), moderate soil drying (MD), and severe soil drying (SD) were imposed during meiosis. MD significantly decreased spikelet degeneration in comparison with WW, due mainly to the alleviation in oxidative damage via enhancing ascorbate-glutathione (AsA-GSH) cycle activity in young panicles, and SD exhibited the opposite effects. Enhanced AsA-GSH cycle strength, decreased oxidative stress, and spikelet degeneration rate were closely associated with the synergistically elevated BR and ABA levels in young panicles in MD. In contrast, low BR and excessive ABA levels led to an increase in spikelet degeneration in SD. The three cultivars exhibited the same tendencies. The intrinsic link among AsA-GSH cycle, oxidative stress, spikelet degeneration rate, and BR and ABA levels was further verified by using transgenic rice lines and chemical regulators. BRs or ABA play a unique role in regulating spikelet degeneration. Synergistically increased BR and ABA levels in MD could work together to strengthen AsA-GSH cycle activity, leading to a reduction in oxidative damage and spikelet degeneration. On the other hand, a severe imbalance between low BR and excessive ABA levels may have contributed to the opposite effects in SD.

Effects of Salt Stress on Grain Yield and Quality Parameters in Rice Cultivars with Differing Salt Tolerance.

Rice yield and grain quality are highly sensitive to salinity stress. Salt-tolerant/susceptible rice cultivars respond to salinity differently. To explore the variation in grain yield and quality to moderate/severe salinity stress, five rice cultivars differing in degrees of salt tolerance, including three salt-tolerant rice cultivars (Lianjian 5, Lianjian 6, and Lianjian 7) and two salt-susceptible rice cultivars (Wuyunjing 30 and Lianjing 7) were examined. Grain yield was significantly decreased under salinity stress, while the extent of yield loss was lesser in salt-tolerant rice cultivars due to the relatively higher grain filling ratio and grain weight. The milling quality continued to increase with increasing levels. There were genotypic differences in the responses of appearance quality to mild salinity. The appearance quality was first increased and then decreased with increasing levels of salinity stress in salt-tolerant rice but continued to decrease in salt-susceptible rice. Under severe salinity stress, the protein accumulation was increased and the starch content was decreased; the content of short branched-chain of amylopectin was decreased; the crystallinity and stability of the starch were increased, and the gelatinization temperature was increased. These changes resulted in the deterioration of cooking and eating quality of rice under severe salinity-stressed environments. However, salt-tolerant and salt-susceptible rice cultivars responded differently to moderate salinity stress in cooking and eating quality and in the physicochemical properties of the starch. For salt-tolerant rice cultivars, the chain length of amylopectin was decreased, the degrees of order of the starch structure were decreased, and pasting properties and thermal properties were increased significantly, whereas for salt-susceptible rice cultivars, cooking and eating quality was deteriorated under moderate salinity stress. In conclusion, the selection of salt-tolerant rice cultivars can effectively maintain the rice production at a relatively high level while simultaneously enhancing grain quality in moderate salinity-stressed environments. Our results demonstrate specific salinity responses among the rice genotypes and the planting of salt-tolerant rice under moderate soil salinity is a solution to ensure rice production in China.

Overlapping peak separation algorithm for ion mobility spectra based on multistrategy JAYA.

RATIONALE: In the field of separation science, ion mobility spectrometry (IMS) plays an important role as an analytical tool. However, the lack of sufficient structural resolution is a common problem in qualitative and quantitative analysis using IMS. A method is needed to solve the problem of overlapping peaks caused by insufficient resolution. METHODS: The method uses multiple strategies to more effectively use population information to balance exploration and exploitation capabilities, prevent local optimization, accurately resolve overlapping peaks, quickly obtain optimal spectral peak model coefficients, and accurately identify compounds. RESULTS: Multistrategy JAYA algorithm's (MSJAYA) performance is compared with improved particle swarm optimization (IPSO), dynamic inertia weight particle swarm optimization (DIWPSO), and multiobjective dynamic teaching-learning-based optimization (MDTLBO). The analysis shows that MSJAYA's maximum separation error is within 0.6%, a level of accuracy not guaranteed by the other algorithms. In addition, the separation error fluctuates within a much smaller range, demonstrating MSJAYA's superior robustness. CONCLUSIONS: Compared with other overlapping peak separation algorithms, MSJAYA is more applicable because no special parameters are used. The method allows fast deconvolution analysis of strong overlapping peaks with multiple components, which greatly improves the resolution of IMS.

Impacts of Inherent Components and Nitrogen Fertilizer on Eating and Cooking Quality of Rice: A Review.

With the continuous improvement of living standards, the preferences of consumers are shifting to rice varieties with high eating and cooking quality (ECQ). Milled rice is mainly composed of starch, protein, and oil, which constitute the physicochemical basis of rice taste quality. This review summarizes the relationship between rice ECQ and its intrinsic ingredients, and also briefly introduces the effects of nitrogen fertilizer management on rice ECQ. Rice varieties with higher AC usually have more long branches of amylopectin, which leach less when cooking, leading to higher hardness, lower stickinesss, and less panelist preference. High PC impedes starch pasting, and it may be hard for heat and moisture to enter the rice interior, ultimately resulting in worse rice eating quality. Rice with higher lipid content had a brighter luster and better eating quality, and starch lipids in rice have a greater impact on rice eating quality than non-starch lipids. The application of nitrogen fertilizer can enhance rice yield, but it also decreases the ECQ of rice. CRNF has been widely used in cereal crops such as maize, wheat, and rice as a novel, environmentally friendly, and effective fertilizer, and could increase rice quality to a certain extent compared with conventional urea. This review shows a benefit to finding more reasonable nitrogen fertilizer management that can be used to regulate the physical and chemical indicators of rice grains in production and to improve the taste quality of rice without affecting yield.

Oxidation kinetics and non-Marcusian charge transfer in dimensionally confined semiconductors.

Electrochemical reactions represent essential processes in fundamental chemistry that foster a wide range of applications. Although most electrochemical reactions in bulk substances can be well described by the classical Marcus-Gerischer charge transfer theory, the realistic reaction character and mechanism in dimensionally confined systems remain unknown. Here, we report the multiparametric survey on the kinetics of lateral photooxidation in structurally identical WS2 and MoS2 monolayers, where electrochemical oxidation occurs at the atomically thin monolayer edges. The oxidation rate is correlated quantitatively with various crystallographic and environmental parameters, including the density of reactive sites, humidity, temperature, and illumination fluence. In particular, we observe distinctive reaction barriers of 1.4 and 0.9 eV for the two structurally identical semiconductors and uncover an unusual non-Marcusian charge transfer mechanism in these dimensionally confined monolayers due to the limit in reactant supplies. A scenario of band bending is proposed to explain the discrepancy in reaction barriers. These results add important knowledge into the fundamental electrochemical reaction theory in low-dimensional systems.

Non-invasive Assessment of Axillary Lymph Node Metastasis Risk in Early Invasive Breast Cancer Adopting Automated Breast Volume Scanning-Based Radiomics Nomogram: A Multicenter Study.

OBJECTIVE: The aim of the work described here was to develop a non-invasive tool based on the radiomics and ultrasound features of automated breast volume scanning (ABVS), clinicopathological factors and serological indicators to evaluate axillary lymph node metastasis (ALNM) in patients with early invasive breast cancer (EIBC). METHODS: We retrospectively analyzed 179 ABVS images of patients with EIBC at a single center from January 2016 to April 2022 and divided the patients into training and validation sets (ratio 8:2). Additionally, 97 ABVS images of patients with EIBC from a second center were enrolled as the test set. The radiomics signature was established with the least absolute shrinkage and selection operator. Significant ALNM predictors were screened using univariate logistic regression analysis and further combined to construct a nomogram using the multivariate logistic regression model. The receiver operating characteristic curve assessed the nomogram's predictive performance. DISCUSSION: The constructed radiomics nomogram model, including ABVS radiomics signature, ultrasound assessment of axillary lymph node (ALN) status, convergence sign and erythrocyte distribution width (standard deviation), achieved moderate predictive performance for risk probability evaluation of ALNs in patients with EIBC. Compared with ultrasound, the nomogram model was able to provide a risk probability evaluation tool not only for the ALNs with positive ultrasound features but also for micrometastatic ALNs (generally without positive ultrasound features), which benefited from the radiomics analysis of multi-sourced data of patients with EIBC. CONCLUSION: This ABVS-based radiomics nomogram model is a pre-operative, non-invasive and visualized tool that can help clinicians choose rational diagnostic and therapeutic protocols for ALNM.

The Improvement of Epothilone D Yield by the Disruption of epoK Gene in Sorangium cellulosum Using TALEN System.

Epothilones are a kind of 16-member macrolides with strong anticancer activity, which was produced by Sorangium cellulosum. Epothlione D shows better drug resistance and safety than taxol in clinical trials. However, the low yield of epothilone D in Sorangium cellulosum and thereof toxicity limited the application of epothilone D. In this study, the epoK gene in gene cluster for epothilone was firstly inactivated by the employment of TALEN gene knockout system. The qRT-PCR analysis and sequencing were performed to confirm the gene deletion of epoK, resulting in the epothilone D yield improvement by 34.9±1.6% and the decrease of epothilone B yield by 34.2±2.5%, which was demonstrated by LC-MS analysis. This study would lay a foundation for the yield improvement of epothilones D, B and thereof derivatives in S. cellulosum by genetic engineering, thus promoting the applications of epothilones in the field of anticancer.

Brassinosteroids mediate moderate soil-drying to alleviate spikelet degeneration under high temperature during meiosis of rice.

This study tested the hypothesis that brassinosteroids (BRs) mediate moderate soil-drying (MD) to alleviate spikelet degeneration under high temperature (HT) stress during meiosis of rice (Oryza sativa L.). A rice cultivar was pot-grown and subjected to normal temperature (NT) and HT treatments during meiosis, and two irrigation regimes including well-watered (WW) and MD were imposed to the plants simultaneously. The MD effectively alleviated the spikelet degeneration and yield loss under HT stress mainly via improving root activity and canopy and panicle traits including higher photosynthetic capacity, tricarboxylic acid cycle activity, and antioxidant capacity than WW. These parameters were regulated by BRs levels in plants. The decrease in BRs levels at HT was due mainly to the enhanced BRs decomposition, and the MD could rescue the BRs deficiency at HT via enhancing BRs biosynthesis and impeding decomposition. The connection between BRs and HT was verified by using rice BRs-deficient mutants, transgenic rice lines, and chemical regulators. Similar results were obtained in the open-air field experiment. The results suggest that BRs can mediate the MD to alleviate spikelet degeneration under HT stress during meiosis mainly via enhancing root activity, canopy traits, and young panicle traits of rice.

IL-17 Imbalance Promotes the Pyroptosis in Immune-Mediated Liver Injury Through STAT3-IFI16 Axis.

Autoimmune hepatitis (AIH) affects all age group and occurs mainly in women. Pyroptosis is a novel programmed cell death featured with cell bursting and release of proinflammatory cytokines. A deeper understanding of AIH pathogenesis will contribute to novel therapy for AIH patients. Here, we aimed to investigate the role of IL-17 in immune-mediated liver injury. The levels of cytokines were measured by ELISA, and mRNA levels of STAT3 and IFN gamma-inducible protein 16 (IFI16) were detected by PCR. Expressions of STAT3, IFI16, gasdermin D and cleaved caspase-1 were measured by western-blotting. Immunohistochemical staining and transmission electron microscopy were applied to evaluate liver histopathological changes of the treated mice. Our results showed that the levels of IFI16 was increased in hepatocytes treated with IL-17 protein, and further elevated after STAT3-overexpressed (STAT3-OE) lentivirus treatment. The levels of IFI16 were reduced in hepatocytes treated with IL-17 neutralizing Ab (nAb), but were significantly increased after STAT3-OE treatment. Pyroptosis was observed in hepatocytes treated with IL-17 protein, and further cell damage was observed after STAT3-OE lentivirus treatment. Liver damage was alleviated in mice treated with IL-17 nAb, however sever damage was experienced after STAT3-OE lentivirus treatment. A binding interaction between IFI16 and STAT3 was detected in IL-17 treated hepatocytes. Glutathione transaminase activity was enhanced in concanavalin A-induced AIH mice compared to the control group (p<0.01). IL-17 plays an important role in activating STAT3 and up-regulating IFI16, which may promote the pyroptosis in AIH-related liver injury through STAT3-IFI16 axis.

[Carbon footprint of major grain crops in the middle and lower reaches of the Yangtze River during 2011-2020]. / 2011­2020年长江中下游地区主要粮食作物生产碳足迹.

The middle and lower reaches of the Yangtze River is one of main grain production areas in China, which is of great significance to food security. Understanding the carbon footprint of major grain crop production is helpful to develop high-yield and low-carbon agriculture. Based on the data of yield, sown area and farmland production input of main grain crops (rice, wheat and maize) in six provinces (Jiangsu, Anhui, Jiangxi, Hubei, Hunan, and Zhejiang) in the middle and lower reaches of the Yangtze River from 2011 to 2020, we estimated carbon footprint in the production of the three grain crops. The results showed that from 2011 to 2020, yield per unit area, planting area, and total yield of rice, wheat and maize were the highest in Jiangsu Province. In terms of area-scaled carbon footprint, rice in the middle and lower reaches of the Yangtze River had the highest area-scaled carbon footprint, with an average of 2.0 t CE·hm-2, followed by wheat and maize. The area-scaled carbon footprint of the three staple crops was increasing. In terms of yield-scaled carbon footprint, rice was the highest, with an average of 0.8 kg CE·kg-1, followed by wheat and maize. In terms of carbon input structure, irrigation electricity, chemical fertilizers and pesticides accounted for a relatively high proportion. Irrigation electricity accounted for 35.0%, 36.3%, and 33.2% of the total carbon input of rice, wheat and maize, respectively. Chemical fertilizers accounted for 28.8%, 32.5%, and 32.5%, respectively, while pesticides accounted for 24.2%, 13.3% and 11.5%, respectively. In terms of carbon efficiency, maize had the highest (3.9 kg·kg-1 CE), followed by rice and wheat. With the green development of agriculture, carbon emission in the production of major grain crops in the middle and lower reaches of the Yangtze River could be reduced by improving irrigation efficiency, fertilizer utilization efficiency, pesticide utilization efficiency and mechanized operation efficiency, as well as diversification of straw returning, cultivation of new varieties and policy leverage.

The Relationships among "STAY-GREEN" Trait, Post-Anthesis Assimilate Remobilization, and Grain Yield in Rice (Oryza sativa L.).

The mobilization and translocation of carbohydrates and mineral nutrients from vegetative plant parts to grains are pivotal for grain filling, often involving a whole plant senescence process. Loss of greenness is a hallmark of leaf senescence. However, the relationship between crop yield and senescence has been controversial for many years. Here, in this study, the overexpression and RNA interference lines of gene of OsNYC3 (Non-Yellow Coloring 3), a chlorophyll catabolism gene, were investigated. Furthermore, exogenous phytohormones were applied, and a treatment of alternate wetting and moderate drying (AWMD) was introduced to regulate the processes of leaf senescence. The results indicated that the delayed senescence of the "STAY-GREEN" trait of rice is undesirable for the process of grain filling, and it would cause a lower ratio of grain filling and lower grain weight of inferior grains, because of unused assimilates in the stems and leaves. Through the overexpression of OsNYC3, application of exogenous chemicals of abscisic acid (ABA), and water management of AWMD, leaf photosynthesis was less influenced, a high ratio of carbohydrate assimilates was partitioned to grains other than leaves and stems as labeled by 13C, grain filling was improved, especially for inferior spikelets, and activities of starch-synthesizing enzymes were enhanced. However, application of ethephon not only accelerated leaf senescence, but also caused seed abortion and grain weight reduction. Thus, plant senescence needs to be finely adjusted in order to make a contribution to crop productivity.

A comprehensive gene expression profile analysis of prostate cancer cells resistant to paclitaxel and the potent target to reverse resistance.

Background: Paclitaxel resistance is the major clinical obstacle in the chemotherapy of prostate cancer (PCa), but the resistant mechanism is less investigated.Purpose: To establish two paclitaxel-resistant PCa cells, provide a comprehensive gene expression profile analysis of resistant cells and the potential target to reverse resistance.Methods: Two Paclitaxel-resistant PCa cells (PC3/PR, LNcap/PR) were established by gradually increasing drug concentration. MTT and transwell assays were performed to detect drug sensitivity, cell proliferation and migration abilities. RNA-Sequencing (RNA-seq) and bioinformatic analyses were performed to identify abnormally expressed genes (AEGs) in resistant cells, and annotate the biological functions of AEGs. The role of the candidate AEG, TLR-4, on the resistant phenotypes was further investigated.Results: The resistance index of resistant cells was 2-3, and they showed a slower proliferation and increased migration ability. 4741 AEGs were screened out (Log2fold change absolute: log2FC(abs) > 1) in the resistant cells, and they were enriched in 2'-5'-oligoadenylate synthetase activity and chemical carcinogenesis. A number of AEGs, CCND2, IGFBP3, FOS, SHH, ZEB2, and members of FGF, FGFR and WNT families were also identified to be involved in cancer- and resistant phenotype-related processes. Finally, TLR-4 was validated significantly increased in resistant cells, and knockdown of TLR-4 increased drug-sensitivity, inhibited the proliferation and migration abilities.Conclusions: The study provided a comprehensive gene expression profile of paclitaxel-resistant PCa cells, and TLR-4 could be a potential target to reverse paclitaxel resistance.

A novel prognostic model based on three clinic-related miRNAs for prostate cancer.

Background: Prostate cancer (PCa) is the second most common malignant tumor in men worldwide. MiRNAs have been reported to play significant roles in prognosis prediction for patients with malignant tumors. Methods: The survival-related miRNAs (sDMIRs) were identified by Cox regression analysis. A risk score model (RSM) was established based on three sDMIRs. The expression levels of sDMIRs in cell lines and clinical samples were detected via quantitative polymerase chain reaction. The correlations between sDMIRs and clinicopathological characteristics of PCa patients were evaluated using the chi-square test and Fisher's exact probability method. Results: Four sDMIRs were remarkably related to the prognosis of PCa patients based on univariate Cox analysis, of which miR-10a-5p, miR-20a-5p, and miR-508-3p were used to establish the RSM. The OS in the low-risk group was better than that in the high-risk group. In the verification of various prostate cell lines and clinical samples from 162 PCa patients, the prominently higher expression of miR-10a-5p and miR-20a-5p and lower expression of miR-508-3p were detected in PCa cell lines and tumor tissues, especially the more advanced T-stage. Besides, the higher expression of miR-20a-5p and miR-10a-5p was significantly correlated to the higher level of PSA, Gleason score, more advanced T-stage, and distant metastasis status. Conclusion: We identify and validate the clinical significance of three sDMIRs and establish a verified RSM to evaluate the prognosis for PCa patients. The findings not only provide a reliable tool for clinical decision-makers to evaluate patients' prognosis but also offer a novel perspective into the field of biomarker identification.

Boosting the performance of single-atom catalysts via external electric field polarization.

Single-atom catalysts represent a unique catalytic system with high atomic utilization and tunable reaction pathway. Despite current successes in their optimization and tailoring through structural and synthetic innovations, there is a lack of dynamic modulation approach for the single-atom catalysis. Inspired by the electrostatic interaction within specific natural enzymes, here we show the performance of model single-atom catalysts anchored on two-dimensional atomic crystals can be systematically and efficiently tuned by oriented external electric fields. Superior electrocatalytic performance have been achieved in single-atom catalysts under electrostatic modulations. Theoretical investigations suggest a universal "onsite electrostatic polarization" mechanism, in which electrostatic fields significantly polarize charge distributions at the single-atom sites and alter the kinetics of the rate determining steps, leading to boosted reaction performances. Such field-induced on-site polarization offers a unique strategy for simulating the catalytic processes in natural enzyme systems with quantitative, precise and dynamic external electric fields.

Tri-n-butyl phosphate delays tissue repair by dysregulating neutrophil function in zebrafish.

Tri-n-butyl phosphate (TnBP) is a widely used organophosphate ester, but its effects on the regenerative process under damaging circumstances remain unknown. In the present study, zebrafish larvae were exposed to 0, 50, 100, 200 and 1000 µg/L TnBP, and the caudal fins were cut at 72 hours post fertilization (hpf). First, after exposure to TnBP, the number of total neutrophils decreased together with decreased neutrophils in the tail, and TnBP inhibited chemotaxis. Second, reactive oxygen species (ROS) levels in the zebrafish decreased greatly. Following exposure to TnBP, transcription levels of many genes regulating fin regeneration, such as fgf20a, fgfr1a, bmp2a and bmp4, were significantly downregulated, while inflammatory factors such as cxcl8a, cxcl18b, il-6, and tnfa were abnormally upregulated. In addition, TnBP inhibited the regenerative area after caudal fin amputation. The inflammatory state was adverse during the regenerative process. In summary, TnBP exposure is immunotoxic and decreases oxidative stress in injured zebrafish larvae.

The sucrose transport regulator OsDOF11 mediates cytokinin degradation during rice development.

Photosynthetic tissues are dynamic structures whose homeostasis depends on the coordination of two antagonistic processes: self-maintenance and supporting sink tissues. The balance of these processes determines plant development, which might be mediated by cytokinin. However, little is known about the link between sucrose transport signaling and cytokinin. Rice (Oryza sativa) DNA BINDING WITH ONE FINGER11 (OsDOF11) is a transcription factor that mediates sucrose transport by inducing the expression of sucrose transporter genes. Here, we found that OsDOF11 loss-of-function mutants showed a semi-dwarf phenotype with a smaller cell length due to increased cytokinin content in source tissues. RNA sequencing and reverse transcription quantitative PCR analyses revealed that genes involved in cytokinin signaling and metabolism were affected in osdof11 mutants. Yeast one-hybrid, dual-luciferase reporter, and chromatin immunoprecipitation experiments showed that OsDOF11 directly binds to the promoter regions of O. sativa CYTOKININ OXIDASE/DEHYDROGENASE4 (OsCKX4). Moreover, mutation of osckx4 in the osdof11 osckx4 double mutant rescued the semi-dwarf phenotype of the osdof11 mutant. Interestingly, exogenous application of kinetin promoted OsDOF11 expression earlier than OsCKX4, and overexpression of O. sativa VIN3-LIKE 2 caused an increase in active cytokinin levels and induced OsDOF11 transcript levels. Taken together, our results suggest a model in which both a sucrose transport regulator (OsDOF11) and cytokinin via OsCKX4 establish a feedback loop to maintain dynamic tissue homeostasis.

Apelin Alleviates Meniscus Endothelial Cell Apoptosis in Osteoarthritis.

Osteoarthritis (OA) is a degenerative disease characterized by articular cartilage and/or chondrocyte destruction, and although it has long been considered as a primary disease, the importance of meniscus endothelial cell modulation in the subchondral microenvironment has recently drawn attention. Previous studies have shown that apelin could potentially inhibit cellular apoptosis; however, it remains unclear whether apelin could play a protective role in protecting the endothelium in the OA meniscus. In this study, with the advantages of single-cell RNA sequencing (scRNA-seq) data, in combination with flow cytometry, we identified two endothelial subclusters in the meniscus, featured by high expression of Homeobox A13 (HOXA13) and Ras Protein-Specific Guanine Nucleotide Releasing Factor 2 (RASGRF2), respectively. Compared with control patients, both subclusters decreased in absolute cell numbers and exhibited downregulated APJ endogenous ligand (APLN, coding for apelin) and upregulated apelin receptor (APLNR, coding apelin receptor). Furthermore, we confirmed that in OA, decreased endothelial cell numbers, including both subclusters, were related to intrinsic apoptosis factors: one more relevant to caspase 3 (CASP3) and the other to BH3-Interacting Domain Death agonist (BID). In vitro culturing of meniscal endothelial cells purified from patients proved that apelin could significantly inhibit apoptosis by downregulating these two factors in endothelial cell subclusters, suggesting that apelin could potentially serve as a therapeutic target for patients with OA.