A transcription-independent mechanism determines rapid periodic fluctuations of BRCA1 expression.

BRCA1 expression is highly regulated to prevent genomic instability and tumorigenesis. Dysregulation of BRCA1 expression correlates closely with sporadic basal-like breast cancer and ovarian cancer. The most significant characteristic of BRCA1 regulation is periodic expression fluctuation throughout the cell cycle, which is important for the orderly progression of different DNA repair pathways throughout the various cell cycle phases and for further genomic stability. However, the underlying mechanism driving this phenomenon is poorly understood. Here, we demonstrate that RBM10-mediated RNA alternative splicing coupled to nonsense-mediated mRNA decay (AS-NMD), rather than transcription, determines the periodic fluctuations in G1/S-phase BRCA1 expression. Furthermore, AS-NMD broadly regulates the expression of period genes, such as DNA replication-related genes, in an uneconomical but more rapid manner. In summary, we identified an unexpected posttranscriptional mechanism distinct from canonical processes that mediates the rapid regulation of BRCA1 as well as other period gene expression during the G1/S-phase transition and provided insights into potential targets for cancer therapy.

Efficacy and Safety of Laser Balloon Versus Irrigated Radiofrequency Ablation as Initial Therapies for Atrial Fibrillation: A Meta-Analysis.

Background: Catheter ablation (CA) is an effective therapy for atrial fibrillation (AF) and, although radiofrequency ablation (RFA) is the standard treatment for pulmonary vein isolation (PVI), it is complex and time-consuming. Laser balloon ablation (LBA) has been introduced to simplify the conventional RFA; however, results of studies comparing LBA and RFA remain controversial. As such, this investigation aimed to comprehensively evaluate the efficacy and safety of LBA versus RFA. Methods: The PubMed, Embase, Cochrane Library, and ClinicalTrials.gov databases were searched for relevant studies. The primary endpoints were the freedom from atrial tachyarrhythmia (ATA) and procedure-related complications. Results: Twelve studies including 1274 subjects were included. LBA and RFA yielded similar rates of freedom from ATA (72.5% vs. 68.7%, odds ratio [OR] = 1.26, 95% confidence interval [CI] 1.0-1.7, p = 0.11) and procedure-related complications (7.7% vs. 6.5%, OR = 1.17, 95% CI 0.72-1.90, p = 0.536). LBA with the second- and third-generation laser balloons (LB2/3) yielded remarkably higher rates of freedom from ATA than RFA using contact-force technology (RFA-CF) (OR = 1.91, p = 0.013). Significantly lower pulmonary vein (PV) reconnection rates (OR = 0.51, p = 0.021), but higher phrenic nerve palsy (PNP) rates (OR = 3.42, p = 0.023) were observed in the LBA group. LBA had comparable procedure (weighted mean difference [WMD] = 8.43 min, p = 0.337) and fluoroscopy times (WMD = 3.09 min, p = 0.174), but a longer ablation time (WMD = 12.57 min, p = 0.00) than those for RFA. Conclusions: LBA and RFA treatments were comparable in terms of freedom from ATA and postprocedural complications in patients with AF. Compared with RFA, LBA was associated with significantly lower PV reconnection rates, but a higher incidence of PNP and longer ablation time.

Extracellular vesicles derived from human Sertoli cells: characterizations, proteomic analysis, and miRNA profiling.

BACKGROUND: Extracellular vesicles (EVs) contain thousands of proteins and nucleic acids, playing an important role in cell-cell communications. Sertoli cells have been essential in the testis as a "nurse cell". However, EVs derived from human Sertoli cells (HSerCs) have not been well investigated. METHODS: EVs were isolated from HSerCs via ultracentrifugation and characterized by transmission electron microscopy, tunable resistive pulse sensing, and Western blotting. The cargo carried by HSerCs-EVs was measured via liquid chromatography-mass spectrometry and GeneChip miRNA Arrays. Bioinformatic analysis was performed to reveal potential functions of HSerCs-EVs. RESULTS: A total of 860 proteins with no less than 2 unique peptides and 88 microRNAs with high signal values were identified in HSerCs-EVs. Biological processes related to molecular binding, enzyme activity, and regulation of cell cycle were significantly enriched. Specifically, many proteins in HSerCs-EVs were associated with spermatogenesis and regulation of immune system, including Septins, Large proline-rich protein BAG6, Clusterin, and Galectin-1. Moreover, abundant microRNAs within HSerCs-EVs (miR-638, miR-149-3p, miR-1246, etc.) had a possible impact on male reproductive disorders such as asthenozoospermia and oligozoospermia. CONCLUSIONS: Our study has shown that HSerCs-EVs contain diverse components such as proteins and microRNAs. Further research is required to evaluate HSerCs-EVs in spermatogenesis, which are underutilized but highly potent resources with particular promise for male infertility.

A Mechanical Modelling and Simulation Method for Resolving PIM Problems in Antennas.

Passive intermodulation (PIM) generated from antennas is a nonlinear distortion phenomenon and causes serious problems to communication quality. Traditional radio frequency (RF) solutions focus on testing the final product to find the PIM source. However, it cannot solve the stability of PIM after the antenna is vibrated. This paper introduces a new method to improve the stability of PIM in the design phase. By studying the mechanism of PIM generation, a simulation method is proposed in this paper by applying mechanical finite element simulation and simulating the structural design of the device under test. Then, the stress at the PIM source is reduced, thereby the PIM stability of the product is improved. This paper adopts this method by studying a typical product, finding the root cause that affects the product PIM magnitude and stability, and optimizing its design. The PIM value of the new scheme is stable by making a prototype and testing. The method provided in this article can effectively improve product development efficiency and assist designers in avoiding the risks of PIM before the product's manufacturing.

Emission characteristics of 99 NMVOCs in different seasonal days and the relationship with air quality parameters in Beijing, China.

In recent years, the haze incidents have occurred frequently in China. Therefore, more attention should be taken in comprehensively determination and analysis of the extended-ranges of volatile organic compounds (VOCs). Here, up to 99 non-methane VOCs (NMVOCs), including not frequently reported partial halocarbons and oxygenated VOC (OVOC) species, were monitored in atmosphere of Beijing. The mean concentration of total NMVOC (TNMVOC) decreased in the order of winter polluted days (216.05 µg m-3) > summer polluted days (127.01 µg m-3) > summer normal days (95.63 µg m-3) > winter normal days (50.25 µg m-3). The ethane to n-butane, ethylene to 1-butene, BTEX, acetaldehyde, acetone, n-hexanal, dichloromethane, chloroform and 1,2-dichloroethane, were determined to be the main composition in their respective alkane, alkene, aromatic, OVOC and halocarbon classes. The minor propylbenzene, diethylbenzene, ethyltoluene, and trimethylbenzene isomer ratios were within the narrow range of 1.3-3.21. Generally, the most abundant NMVOCs were alkanes in winter but OVOCs in summer. TNMVOC significantly positively correlated with PM10, PM2.5, CO, RH, SO2 (winter), NO2 (winter), but negatively with windspeed, SSD and PRS (winter). The opposite correlation was observed between TNMVOC and O3 in winter and summer. There was no meaningful correlation between TNMVOC and T, PRS (summer), SO2 (summer) and NO2 (summer). 3D surface graphs, built by MATLAB, were drawn to investigate the relationship between PM2.5 and NMVOC taking air quality parameters into account. The PM2.5 concentration increased non-linearly as TNMVOC concentration increased, with various surface graphs. Unlike other air quality parameters, O3 affected the relationship differently between winter and summer. The findings presented herein may provide a new train of thought for occurrence of haze.

Thermal Oxidation Degradation of 2,2',4,4'-Tetrabromodiphenyl Ether over LiαTiOx Micro/Nanostructures with Dozens of Oxidative Product Analyses and Reaction Mechanisms.

Flowerlike LiαTiOx micro/nanostructures were successfully synthesized to degrade 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) at 250-350 °C. The pseudo-first-order kinetics rate constant of the reaction at 300 °C was in the range of 0.034-0.055 min-1. The activation energy was as low as 39.9-48.1 kJ/mol. The excellent performance attained over LiαTiOx was attributed to Li dopant having the electron-donating effect, which enhanced the oxygen species mobility. The oxidative reaction was believed to be the dominant degradation pathway following the Mars-van Krevelen mechanism, being accompanied by the weak hydrodebromination occurrence generating the trace mono- to tri-BDEs. More than 70 types of oxidation products containing diphenyl ether backbone, single-benzene rings, and ring-opened products were detected by GC-MS with derivatization, ESI-FT-ICR-MS, and ion chromatography. An increase in the number of ring-cracked oxidative products under prolonged reaction was observed by ESI-FT-ICR-MS analysis according to the van Krevelen diagram. In the oxidative reaction, a series of oxidative products, such as OH-tri-BDEs and OH-tetra-BDEs, first formed via the nucleophilic O2- attack and subsequently transformed into dibromophenol, tribromophenol, and benzenedicarboxylic and benzoic acids, etc. They could be further attacked by electrophilic O2- and O- and completely cracked to small molecules such as formic, acetic, propionic, and butyric acids.

[Nonlinear Variations in PM2. 5 Concentration in the Three Major Urban Agglomerations in China].

ITA and Beast methods were used to quantitatively analyze the nonlinear process of a PM2.5 concentration time series based on the PM2.5 concentration data of the three major urban agglomerations in China. The results showed that： ① the degree of the PM2.5 pollution in the three major urban agglomerations had decreased， and the high-concentration areas had noticeably shrunk. The degree of spatial polarization of PM2.5 concentration was reduced， and the spatial difference was narrowed. The PM2.5 concentration in most areas showed downward trends， but the degree of change was not the same. Compared with the YRD and PRD， the concentration of PM2.5 in the BTH was still at a relatively high level. ② The concentration of PM2.5 in the three major urban agglomerations had seasonal variation characteristics that were high in winter and spring and low in summer and autumn. There were obvious differences in PM2.5 concentration between winter and summer， and the convergence of PM2.5 concentration in summer was greater than that in winter. Areas with high PM2.5 concentration also had obvious downward trends， but the downward trends of PM2.5 concentration in the PRD were not obvious compared with those in the YRD and BTH. ③ The PM2.5 concentration time series of the three major urban agglomerations all had significant downward trends： Beijing-Tianjin-Hebei （BTH） > the Yangtze River Delta （YRD） > the Pearl River Delta （PRD）. The PM2.5 concentration had the largest downward trends in winter. The higher the PM2.5 pollution level， the greater the downward trends. ④ The trend component of the PM2.5 concentration time series in the BTH had two change points， and there was one change point in the seasonal component. The trend and seasonal components of the PM2.5 concentration time series in the YRD had no change point. There was no change point in the seasonal component but one change point in the trend component of the PM2.5 concentration time series in the PRD. These results can provide scientific references for regional air pollution control.

Recognition of screening out hierarchical toxic contaminants tuned by quantified pseudo-components from complex engineering co-combustion.

Co-combustion of industrial and municipal solid wastes has emerged as the most promising disposal technology, yet its effect on unknown contaminants generation remains rarely revealed due to waste complexity. Hence, six batches of large-scale engineering experiments were designed in an incinerator of 650 t/d, which overcame the inauthenticity and deviation of laboratory tests. 953-1772 non-targeted compounds were screened in fly ash. Targeting the impact of co-combustion, a pseudo-component matrix model was innovatively integrated to quantitatively extract nine components from complex wastes grouped into biomass and plastic. Thus, the influence was evaluated across eight dimensions, covering molecular characteristics and toxicity. The effect of co-combustion with biomass pseudo-components was insignificant. However, co-combustion with high ratios of plastic pseudo-components induced higher potential risks, significantly promoting the formation of unsaturated hydrocarbons, highly unsaturated compounds (DBE≥15), and cyclic compounds by 19 %- 49 %, 17 %- 31 %, and 7 %- 27 %, respectively. Especially, blending with high ratios of PET plastic pseudo-components produced more species of contaminants. Unique 2 Level I toxicants, bromomethyl benzene and benzofuran-2-carbaldehyde, as well as 4 Level II toxicants, were locked, receiving no concern in previous combustion. The results highlighted risks during high proportion plastics co-combustion, which can help pollution reduction by tuning source wastes to enable healthy co-combustion.

A bibliometric analysis of efferocytosis in cardiovascular diseases from 2001 to 2022.

INTRODUCTION: In recent years, efferocytosis in cardiovascular diseases has become an intense area of research. However, only a few bibliometric analyses have been conducted in this area. In this review, we used CiteSpace 5.7. R2 and VOSviewer 1.6.17 software to perform text mining and knowledge map analysis. This study summarizes the latest progress, development paths, frontier research hotspots, and future research trends in this field. MATERIALS AND METHODS: Studies on efferocytosis in cardiovascular diseases were downloaded from the Web of Science Core Collection. RESULTS: In total, 327 studies published by 506 institutions across 42 countries and regions were identified. The number of studies on efferocytosis in cardiovascular diseases has increased over time. Arteriosclerosis Thrombosis and Vascular Biology published the highest number of articles and was the top co-cited journal. Tabas Ira. was the most prolific researcher and co-cited the most. The most productive countries were the United States and China. Columbia University, Harvard Medical School, and Brigham Women's Hospital were the 3 most productive institutions in the field of research. Keyword Co-occurrence, Clusters, and Burst analyses showed that inflammation, atherosclerosis, macrophages, and phagocytosis appeared with the highest frequency in these studies. CONCLUSION: Multinational cooperation and multidisciplinary intersections are characteristic trends of development in the field, and the immune microenvironment, glycolysis, and lipid metabolism will be the focus of future research.

SPSI: A Novel Composite Index for Estimating Panicle Number in Winter Wheat before Heading from UAV Multispectral Imagery.

Rapid and accurate estimation of panicle number per unit ground area (PNPA) in winter wheat before heading is crucial to evaluate yield potential and regulate crop growth for increasing the final yield. The accuracies of existing methods were low for estimating PNPA with remotely sensed data acquired before heading since the spectral saturation and background effects were ignored. This study proposed a spectral-textural PNPA sensitive index (SPSI) from unmanned aerial vehicle (UAV) multispectral imagery for reducing the spectral saturation and improving PNPA estimation in winter wheat before heading. The effect of background materials on PNPA estimated by textural indices (TIs) was examined, and the composite index SPSI was constructed by integrating the optimal spectral index (SI) and TI. Subsequently, the performance of SPSI was evaluated in comparison with other indices (SI and TIs). The results demonstrated that green-pixel TIs yielded better performances than all-pixel TIs apart from TI[HOM], TI[ENT], and TI[SEM] among all indices from 8 types of textural features. SPSI, which was calculated by the formula DATT[850,730,675] + NDTICOR[850,730], exhibited the highest overall accuracies for any date in any dataset in comparison with DATT[850,730,675], TINDRE[MEA], and NDTICOR[850,730]. For the unified models assembling 2 experimental datasets, the RV2 values of SPSI increased by 0.11 to 0.23, and both RMSE and RRMSE decreased by 16.43% to 38.79% as compared to the suboptimal index on each date. These findings indicated that the SPSI is valuable in reducing the spectral saturation and has great potential to better estimate PNPA using high-resolution satellite imagery.

Genome-wide analysis of the MADS-box gene family involved in salt and waterlogging tolerance in barley (Hordeum vulgare L.).

MADS-box transcription factors are crucial members of regulatory networks underlying multiple developmental pathways and abiotic stress regulatory networks in plants. Studies on stress resistance-related functions of MADS-box genes are very limited in barley. To gain insight into this gene family and elucidate their roles in salt and waterlogging stress resistance, we performed genome-wide identification, characterization and expression analysis of MADS-box genes in barley. A whole-genome survey of barley revealed 83 MADS-box genes, which were categorized into type I (Mα, Mß and Mγ) and type II (AP1, SEP1, AGL12, STK, AGL16, SVP and MIKC*) lineages based on phylogeny, protein motif structure. Twenty conserved motifs were determined and each HvMADS contained one to six motifs. We also found tandem repeat duplication was the driven force for HvMADS gene family expansion. Additionally, the co-expression regulatory network of 10 and 14 HvMADS genes was predicted in response to salt and waterlogging stress, and we proposed HvMADS11,13 and 35 as candidate genes for further exploration of the functions in abiotic stress. The extensive annotations and transcriptome profiling reported in this study ultimately provides the basis for MADS functional characterization in genetic engineering of barley and other gramineous crops.

[Scale Dependence Between PM2.5 and Meteorological Factors and Its Influencing Factors in "2+26" Cities].

Based on the PM2.5 concentration and meteorological data of "2+26" cities, the variations in PM2.5 time series were analyzed by the continuous wavelet transform(CWT) and discrete wavelet transform(DWT). Wavelet coherence(WTC) and multiple wavelet coherence(MWC) were used to quantify the response relationship between PM2.5 and single/multiple meteorological factors in the time-frequency domain. Partial wavelet coherence(PWC) was used to quantitatively evaluate the influence of atmospheric teleconnection factors on the response relationship. The results showed that:① the concentration of PM2.5 in the "2+26" cities had the spatial distribution characteristics of high in the middle area and low in the peripheral area. The PM2.5 mutation events were mainly concentrated before 2018 and mostly occurred in winter when the meteorological conditions were stable. The annual scale period of 256-512 d was relatively stable, and it was also the dominant period of the PM2.5 time series. ② The coherences between PM2.5 and meteorological factors depended on the time-frequency scale and variable combination. At all time-frequency scales, PM2.5 had strong coherences with relative humidity and temperature. At small and medium time-frequency scales, PM2.5 had strong coherences with wind speed. At large scales, PM2.5 had strong coherences with temperature. The combination of precipitation, temperature, and relative humidity could explain the variation in PM2.5 at all time-frequency scales. ③ At different time-frequency scales, the enhancement/weakening effects of atmospheric teleconnection factors on the response relationship were not the same. At all time-frequency scales, the El Niño-Southern Oscillation(ENSO) had a greater impact on the response relationship between PM2.5 and precipitation/temperature, and the Pacific decadal oscillation(PDO) had a greater impact on the response relationship between PM2.5 and relative humidity/wind speed. These results provide reference for regional air pollution control.

In situ mapping of ion distribution profiles and gene expression reveals interactions between hypoxia and Mn2+/Fe2+ availability in barley roots.

Flooding stress affects soil properties thus altering the availability, uptake, and transport of mineral nutrients in plant roots. Flooding stress also increases the amount of soluble Mn2+ and Fe2+ in the soil and their uptake by plants, causing elemental toxicity. However, as oxygen profiles in plant roots are not uniform, it is still unclear how soil flooding will affect Mn2+/Fe2+ absorption and distribution in different cell types and tissues. In this study, waterlogging sensitive barley variety NasoNijo (NN) and tolerant variety TX9425 (TX) were exposed to hypoxia, metal (Mn2+ and Fe2+), and combined hypoxia + metal treatment to map the in situ ion profiles at different regions of barley root. We found that combined hypoxia and metal stress causes significantly more reduction in plant biomass compared with the single submergence or metal stress. Despite this, more Fe and Mn were accumulated under metal stress condition than those under combined stress, regardless of variety. Cultivar NN absorbed more Fe and Mn than TX in the cortical cells of the root meristem and in the mature zone under metal stress which was also verified by histochemical detection. In the mature zone, the expressions of Fe and Mn transporter genes including HvADPRibase-Mn (Manganese-dependent ADP-ribose), HvZIP1 (zinc-regulated transporter /Fe-regulated transporter-like protein 1), HvYS1 (yellow stripe 1), HvNRAMP5 (Natural Resistance-Associated Macrophage Protein 5) were significantly downregulated under all three treatments in both barley varieties except HvADPRibase-Mn HvZIP1 cortex of TX were unchanged under metal stress. Interestingly, the transcripts of HvMTP1 (metal tolerance protein 1) were significantly downregulated by metal and combined stress in stele and upregulated by hypoxia and metal stress in cortex of TX, but not affected in NN. It is concluded that Fe and Mn absorption involving HvMTP1is associated with the extent of waterlogging tolerance in barley.

Comparison of two novel methods for counting wheat ears in the field with terrestrial LiDAR.

BACKGROUND: The metrics for assessing the yield of crops in the field include the number of ears per unit area, the grain number per ear, and the thousand-grain weight. Typically, the ear number per unit area contributes the most to the yield. However, calculation of the ear number tends to rely on traditional manual counting, which is inefficient, labour intensive, inaccurate, and lacking in objectivity. In this study, two novel extraction algorithms for the estimation of the wheat ear number were developed based on the use of terrestrial laser scanning (TLS) in conjunction with the density-based spatial clustering (DBSC) algorithm based on the normal and the voxel-based regional growth (VBRG) algorithm. The DBSC involves two steps: (1) segmentation of the point clouds using differences in the normal vectors and (2) clustering of the segmented point clouds using a density clustering algorithm to calculate the ear number. The VBRG involves three steps: (1) voxelization of the point clouds, (2) construction of the topological relationships between the voxels as a connected region using the k-dimensional tree, and (3) detection of the wheat ears in the connected areas using a regional growth algorithm. RESULTS: The results demonstrated that DBSC and VBRG were promising in estimating the number of ears for different cultivars, planting densities, N fertilization rates, and growth stages of wheat (RMSE = 76 ~ 114 ears/m2, rRMSE = 18.62 ~ 27.96%, r = 0.76 ~ 0.84). Comparing the performance of the two algorithms, the overall accuracy of the DBSC (RMSE = 76 ears/m2, rRMSE = 18.62%, r = 0.84) was better than that of the VBRG (RMSE = 114 ears/m2, rRMSE = 27.96%, r = 0.76). It was found that with the DBSC, the calculation in points as units permitted more detailed information to be retained, and this method was more suitable for estimation of the wheat ear number in the field. CONCLUSIONS: The algorithms adopted in this study provide new approaches for non-destructive measurement and efficient acquisition of the ear number in the assessment of the wheat yield phenotype.

Transcriptome analysis highlights the role of ferroptosis in palmitic acid-induced endothelial dysfunction.

Background: Palmitic acid (PA) has a lipotoxic effect on blood vessels, leading to endothelial dysfunction and cell death. The underlying mechanisms are not yet fully understood. Aim: We sought to investigate the effects of PA on endothelial cells, with an emphasis on ferroptosis. Methods: Rat corpus cavernosum endothelial cells (RCCECs) and human umbilical vein endothelial cells (HUVECs) were treated with PA to induce a pattern of cell death, as evidenced by the evaluation of cell viability. The differentially expressed genes were measured via RNA sequencing to reveal potential mechanisms. The intracellular levels of glutathione (GSH), malondialdehyde (MDA), ferrous ion (Fe2+), and reactive oxygen species (ROS) were evaluated using commercial kits. Western blot was performed to determine the expressions of relative proteins. Outcomes: At the end of the study period, the evaluated outcomes were cell viability, transcriptome profiles, the expressions of glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11), as well as levels of GSH, MDA, Fe2+, and ROS. Results: PA-induced cell death of RCCECs and HUVECs was demonstrated in a dose- and time-dependent manner. Based on the findings of RNA-sequencing (RNA-seq), enrichment of many biological processes associated with cell cycle and response to stimulus occurred. More importantly, ferroptosis was highlighted in the bioinformatic analysis of both endothelial cells. The levels of intracellular Fe2+, MDA, and ROS were significantly increased following PA exposure while GSH was decreased, suggesting excessive iron accumulation, development of lipid peroxidation, and imbalanced redox homeostasis. Mechanistically, PA decreased the protein expression levels of GPX4 and SLC7A11 in endothelial cells, both of which played crucial roles in ferroptotic cell death. Clinical Translation: This study suggests that ferroptosis may be a useful target for novel therapeutic interventions for endothelial dysfunction and cell death in vascular diseases such as erectile dysfunction. Strengths and Limitations: In this study, we found that ferroptosis could participate in PA-induced endothelial dysfunction and cell death. A limitation of the study is that it did not shed light on the overall mechanisms of this process. Therefore, further research on the intricate networks of regulating ferroptosis is needed. Conclusion: Overall, the occurrence of ferroptosis was demonstrated in the PA-treated HUVECs and RCCECs in this study.

Icariside II induces rapid phosphorylation of endothelial nitric oxide synthase via multiple signaling pathways.

Icariside II, as a favonoid compound derived from epimedium, has been proved to involed in a variety of biological and pharmacological effects such as anti-inflammatory, anti-osteoporosis, anti-oxidation, anti-aging, and anti-cancer but its mechanism is unclear, especially in terms of its effect on post-transcriptional modification of endothelial nitric oxide synthase (eNOS). Phosphorylation of eNOS plays an important role in the synthesis of nitric oxide in endothelial cells, which is closely related to erectile dysfunction, atherosclerosis, Alzheimer's disease, and other diseases. Our study aims to investigate the effect and mechanism of Icariside II on the rapid phosphorylation of eNOS. In this study, human umbilical vein endothelial cells (HUVECs) were stimulated with Icariside II in the presence or absence of multiple inhibitors (1 µM), including LY294002 (PI3K-inhibitor), MK-2206 (AKT-inhibitor), Bisindolylmaleimide X (AMPK-inhibitor), H-89 (CaMKII-inhibitor), KN-62 (PKA-inhibitor), Dorsomorphin (PKC-inhibitor). The proliferation of HUVECs was assessed using cell counting kit-8 (CCK-8). The release of nitric oxide (NO) within HUVECs was detected via fluorescence probe (DAF-FM). Western blot was used to examine the effect of Icariside II on the expression of eNOS, phosphorylation of eNOS, and common signaling pathways proteins. In this study, Icariside II was found to promote the cell proliferation and rapid NO release in HUVECs. The phosphorylation of eNOS-Ser1177 was significantly increased after Icariside II stimulation and reached a peak at 10 min (p < 0.05). Meanwhile, the phosphorylation of eNOS-Thr495 was significantly decreased after 45 min of stimulation (p < 0.05). Following the intervention with multiple inhibitors, it was found that MK-2206 (AKT inhibitor), LY294002 (PI3K inhibitor), KN-62 (AMPK inhibitor), and Bisindolylmaleimide X (PKC inhibitor) could significantly inhibit the phosphorylation of eNOS-Ser1177 caused by Icariside II (p < 0.05), while MK-2206, LY294002, and Bisindolylmaleimide X reversed the alleviated phosphorylation of eNOS-Thr495. We concluded that Icariside can regulate rapid phosphorylation of eNOS- Ser1177 and eNOS-Thr495 via multiple signaling pathways, resulting in the up-regulation of eNOS and the increased release of NO.

Proteomic analysis and miRNA profiling of human testicular endothelial cell-derived exosomes: the potential effects on spermatogenesis.

Testicular endothelial cells have been found to play an important role in spermatogenesis and fertility, but their mechanism is obscure. Exosomes released by various cells are recognized as cell-cell communication mediators during the initiation and progression of many diseases. Therefore, the current study aimed to investigate the protein and miRNA components of human testicular endothelial cell-derived exosomes (HTEC-Exos) and to explore their potential effects on spermatogenesis. In this study, HTEC-Exos were first isolated by the ultracentrifugation method, and then identified by nanoparticle tracking analysis, transmission electron microscopy (TEM), and western blotting. The characteristics of HTEC-Exos were examined by liquid chromatography-mass spectrometry and microRNA (miRNA) chip analysis. Bioinformatics analysis was performed to explore the potential role of the exosomal content on spermatogenesis. A total of 945 proteins were identified, 11 of which were closely related to spermatogenesis. A total of 2578 miRNAs were identified. Among them, 30 miRNAs demonstrated potential associations with male reproductive disorders, such as azoospermia, and spermatogenesis disorders. In particular, 11 out of these 30 miRNAs have been proven to be involved in spermatogenesis based on available evidence. This study provides a global view of the proteins and miRNAs from HTEC-Exos, suggesting that HTEC-Exos may function as potential effectors during the process of spermatogenesis.

Icariside â ¡ Attenuates Palmitic Acid-Induced Endothelial Dysfunction Through SRPK1-Akt-eNOS Signaling Pathway.

Background: Endothelial dysfunction is commonly accompanied by a reduced capacity for nitric oxide (NO) production and decreased NO sensitivity, playing a central role in numerous vascular diseases. Saturated free fatty acids are known to reduce NO production and then induce endothelial dysfunction. Alternative splicing participates in the regulation of cellular and tissular homeostasis and is highly regulated by serine-arginine protein kinase (SRPK1). The role of SRPK1 in the biology of endothelial cells remains elusive. Icariside Ⅱ (ICA Ⅱ) has been reported to have protective effects on endothelial function. However, the specific molecular mechanisms are still unknown. The purpose of this study is to explore the role of SRPK1 in the biology of endothelial cells and the underlying mechanism of ICA Ⅱ on palmitic acid (PA) induced endothelial dysfunction. Methods: Endothelial dysfunction was induced using PA in human umbilical vein endothelial cells (HUVECs). The expression and phosphorylation of related proteins in the SRPK1-Akt-eNOS signaling pathway were detected by Western Blot. Cell Counting Kit-8 assay and Ki-67 immunofluorescence were used to estimate cell viability. Endothelial cell function was assessed by detecting NO production using DAF-FM DA. Interaction between ICA Ⅱ and SRPK1 was demonstrated by a biotinylated protein interaction pull-down assay. Results: The expressions of eNOS, Akt, and SRPK1 were down-regulated in the endothelial dysfunction stimulated by PA. SRPK1 inhibitor SPHINX31 restrained endothelial cell viability in a dose-dependent manner. Moreover, inhibition of SRPK1 using SPHINX31 and knockdown of SRPK1 by shRNA also showed a down-regulation of the proteins associated with the SRPK1-Akt-eNOS signaling pathway. Biotinylated protein interaction pull-down assay revealed that ICA Ⅱ could be directly bound with SRPK1. On the other hand, ICA Ⅱ could attenuate the PA-induced endothelial dysfunction and restore cell viability through the SRPK1-Akt-eNOS pathway. Conclusions: ICA Ⅱ, bound with SRPK1, could attenuate the endothelial dysfunction induced by the PA in HUVECs via the SRPK1-Akt-eNOS signaling pathway.

Single-Cell RNA Sequencing of Human Corpus Cavernosum Reveals Cellular Heterogeneity Landscapes in Erectile Dysfunction.

Purpose: To assess the diverse cell populations of human corpus cavernosum in patients with severe erectile dysfunction (ED) at the single-cell level. Methods: Penile tissues collected from three patients were subjected to single-cell RNA sequencing using the BD Rhapsody™ platform. Common bioinformatics tools were used to analyze cellular heterogeneity and gene expression profiles from generated raw data, including the packages Seurat, Monocle, and CellPhoneDB. Results: Disease-related heterogeneity of cell types was determined in the cavernous tissue such as endothelial cells (ECs), smooth muscle cells, fibroblasts, and immune cells. Reclustering analysis of ECs identified an arteriole ECs subcluster and another one with gene signatures of fibroblasts. The proportion of fibroblasts was higher than the other cell populations and had the most significant cellular heterogeneity, in which a distinct subcluster co-expressed endothelial markers. The transition trajectory of differentiation from smooth muscle cells into fibroblasts was depicted using the pseudotime analysis, suggesting that the expansion of corpus cavernosum is possibly compromised as a result of fibrosis. Cell-cell communications among ECs, smooth muscle cells, fibroblasts, and macrophages were robust, which indicated that inflammation may also have a crucial role in the development of ED. Conclusions: Our study has demonstrated a comprehensive single-cell atlas of cellular components in human corpus cavernosum of ED, providing in-depth insights into the pathogenesis. Future research is warranted to explore disease-specific alterations for individualized treatment of ED.

Phytoremediation of antibiotic-contaminated wastewater: Insight into the comparison of ciprofloxacin absorption, migration, and transformation process at different growth stages of E. crassipes.

The selection of aquatic plants at different growth stages and their absorption, migration, and transformation mechanisms has yet to be clarified. In this study, Eichhornia crassipes at the seedling and mature stages were selected to uptake antibiotics under hydroponic conditions. The results showed that the enrichment of ciprofloxacin (CIP) in roots at the seedling and mature stages were 7.72～2114.39 µg g-1 and 0.07～3711.33 µg g-1, respectively. The enrichment of CIP in aerial parts at the seedling and mature stages were 16.38～24.24 µg g-1 and 9.55～20.13 µg g-1, respectively. The translocation from roots to aerial parts at the seedling stage was high, as evidenced by the relatively higher transfer factor (TF). In addition, eight and ten major metabolic products were observed in the tissues of seeding and mature stage of E. crassipes, respectively. The metabolic pathway of CIP was short at the maturity stage, and CIP had a strong upward migration ability at the seedling stage, facilitating long-time photodegradation. However, E. crassipes exhibited a poor CIP tolerance at the mature stage and decayed relatively early. Therefore, the seedling stage of E. crassipes was proposed to be applied for phytoremediation, and these findings might improve the ability to phytoremediation of antibiotic-contaminated water.