The male reproductive toxicity after 5-Fluorouracil exposure: DNA damage, oxidative stress, and mitochondrial dysfunction in vitro and in vivo.

5-Fluorouracil (5-FU), a chemotherapeutic drug used to treat a variety of cancers, can enter the environment through different routes, causing serious public health and environmental concerns. It has been reported that 5-FU exposure adversely affects male reproductive function, and its effects on this system cannot be avoided. In this study, using western blotting and quantitative polymerase chain reaction studies, we found that 5-FU promoted testicular injury by inducing oxidative stress, which was accompanied by the inhibition of nuclear factor erythroid 2-related factor 2/antioxidant response element signaling. Accumulation of reactive oxygen species (ROS) aggravated 5-FU-mediated mitochondrial dysfunction and apoptosis in murine cell lines and testes, indicating oxidative stress and mitochondrial-dependent apoptotic signaling play crucial roles in the damage of spermatogenic cells caused. N-Acetyl-L-cysteine, an antioxidant that scavenges intracellular ROS, protected spermatogenic cells from 5-FU-induced oxidative damage and mitochondrial dysfunction, revealing the important role of ROS in testicular dysfunction caused by 5-FU. We found that 5-FU exposure induces testicular cell apoptosis through ROS-mediated mitochondria pathway in mice. In summary, our findings revealed the reproductive toxicological effect of 5-FU on mice and its mechanism, provided basic data reference for adverse ecological and human health outcomes associated with 5-FU contamination or poisoning.

pparß regulates lipid catabolism by mediating acox and cpt-1 genes in Scophthalmus maximus under heat stress.

Peroxisome proliferator-activated receptor ß (pparß) is a key gene-regulating lipid metabolism pathway, but its function in turbot remains unclear. In this study, the CDS of pparß was cloned from kidney for the first time. The CDS sequence length was 1533 bp encoding 510 amino acids. Structural analysis showed that the pparß protein contained a C4 zinc finger and HOLI domain, suggesting that the pparß gene of turbot has high homology with the PPAR gene of other species. The high expression patterns of pparß, acox, and cpt-1 at high temperatures, as shown through qPCR, indicated that high temperatures activated the transcriptional activity of pparß and increased the activity of the acox and cpt-1 genes. The expression of acox and cpt-1 was significantly inhibited when pparß was downregulated using RNAi technology and inhibitor treatments, suggesting that pparß positively regulated acox and cpt-1 expression at high temperatures and, thus, modulates lipid catabolism activity. These results demonstrate that pparß is involved in the regulation of lipid metabolism at high temperatures and expand a new perspective for studying the regulation of lipid metabolism in stress environments of teleost.

Emerging roles of ferroptosis in male reproductive diseases.

Ferroptosis is a type of programmed cell death mediated by iron-dependent lipid peroxidation that leads to excessive lipid peroxidation in different cells. Ferroptosis is distinct from other forms of cell death and is associated with various diseases. Iron is essential for spermatogenesis and male reproductive function. Therefore, it is not surprising that new evidence supports the role of ferroptosis in testicular injury. Although the molecular mechanism by which ferroptosis induces disease is unknown, several genes and pathways associated with ferroptosis have been linked to testicular dysfunction. In this review, we discuss iron metabolism, ferroptosis, and related regulatory pathways. In addition, we analyze the endogenous and exogenous factors of ferroptosis in terms of iron metabolism and testicular dysfunction, as well as summarize the relationship between ferroptosis and male reproductive dysfunction. Finally, we discuss potential strategies to target ferroptosis for treating male reproductive diseases and provide new directions for preventing male reproductive diseases.

Deep-learning based segmentation of ultrasound adipose image for liposuction.

BACKGROUND: To develop an automatic and reliable ultrasonic visual system for robot- or computer-assisted liposuction, we examined the use of deep learning for the segmentation of adipose ultrasound images in clinical and educational settings. METHODS: To segment adipose layers, it is proposed to use an Attention Skip-Convolutions ResU-Net (Attention SCResU-Net) consisting of SC residual blocks, attention gates and U-Net architecture. Transfer learning is utilised to compensate for the deficiency of clinical data. The Bama pig and clinical human adipose ultrasound image datasets are utilized, respectively. RESULTS: The final model obtains a Dice of 99.06 ± 0.95% and an ASD of 0.19 ± 0.18 mm on clinical datasets, outperforming other methods. By fine-tuning the eight deepest layers, accurate and stable segmentation results are obtained. CONCLUSIONS: The new deep-learning method achieves the accurate and automatic segmentation of adipose ultrasound images in real-time, thereby enhancing the safety of liposuction and enabling novice surgeons to better control the cannula.

A framework for precisely thinning planning in a managed pure Chinese fir forest based on UAV remote sensing.

Where and how many trees should be thinned in a pure managed forest to improve forest quality and increase ecological benefits are important forest questions. In this study we address such challenges by providing a novel framework for planning thinning operations through Unmanned Aerial Vehicle (UAV) remote sensing techniques, which can not only obtain forest attributes of its entire stand with spatial properties, but also optimize the selection of thinning areas, thinning intensities and cut-trees. This study helps to reduce the costs of time-consuming and laborious ground investigations. The framework was demonstrated by applying it into a subtropical Chinese fir plantation in southeastern China. Results showed that RGB images acquired by a low-cost UAV have great potential in depicting forest structure. The overall accuracy of the individual tree detection in the case study was 85.19 % ± 0.48 %. The overall accuracy and the intersection over union of the non-crown area extraction were 94.94 % and 82.65 %, respectively. For the two determined thinning areas, 19.5 % and 14.3 % crown density were required to thin in the primary and secondary regions, respectively. In addition, the top-down perspective of UAV remote sensing makes up for the limitations of the bottom-up perspective of traditional forestry. The framework can act as a basic model for forest managers to modify and expand for customizing detailed thinning guidelines.

Vertical distributions of atmospheric HONO and the corresponding OH radical production by photolysis at the suburb area of Shanghai, China.

Nitrous acid (HONO) is considered as one of the main sources of the hydroxyl radical (OH), the most relevant oxidant in the atmosphere. Multi-AXis-Differential Optical Absorption Spectroscopy (MAX-DOAS) measurements were conducted to obtain the vertical profiles of aerosol and HONO from November 1, 2020 to January 31, 2021 at a suburb site of Shanghai, China. HONO was mainly distributed near the surface, but high values HONO occasionally occurred around 0.7 km, indicating an unaccounted source of daytime HONO at high altitudes. The positive correlation between HONO and aerosols suggested that the photo-enhanced heterogeneous reactions on the aerosol surface were an important source of daytime HONO at high altitudes. To obtain the vertical distribution of OH production by HONO photolysis (P(OH)HONO), the vertical profiles of photolysis rate of HONO (JHONO) were calculated by establishing a method of combining observations with empirical relationship based on heterogeneous atmospheric and radiative transfer models. The JHONO increased approximately linearly with increasing altitudes and the noontime averages value of JHONO near the ground were 6.68 × 10-4 s-1, which was strongly negatively affected by aerosols in the morning and afternoon. The P(OH)HONO profile varied in different months (November, December, January) that the changes were mainly affected by HONO and JHONO. P(OH)HONO was more positively affected by JHONO at high altitude and noon but greatly influenced by HONO concentrations in the morning and afternoon.

Genetic Mechanism for Antioxidant Activity of Endogenous Enzymes under Salinity and Temperature Stress in Turbot (Scophthalmus maximus).

Three antioxidant properties (corresponding to the enzymes superoxide dismutase (SOD), catalase, and glutathione peroxidase) were measured in the livers of Scophthalmus maximus under different salinities/temperatures (5, 10, 20, 30, and 40‱/17, 20, 23, 25, and 28 °C). Split-plot (SP) analysis, additive main effects, and multiplicative interaction (AMMI) and genotype × environment interaction (GGE) biplots were used to analyze genotype × salinity/temperature interactions for antioxidant properties. The results of the SP analysis show that the activity of the three antioxidant properties was significantly (p < 0.01) affected by salinity/temperature, antioxidant, and salinity/temperature × antioxidant interaction. The results of the AMMI analysis for salinity reveal that the effect of genotype, salinity, and genotype × salinity interaction on antioxidant properties reached a significant level (p < 0.001); 92.1065%, 2.6256%, and 4.4360% of the total sum of squares for antioxidant property activity were attributable to the effects of genotype, salinity, and genotype × salinity interaction, respectively. The results of GGE biplot analysis for salinity reveal differences in the activity ranking of the three antioxidant properties under five salinities; this difference expands with an decrease or increase in salinity from 30‱ (optimum salinity). A salinity of 5‱ had the strongest ability to identify the three antioxidant properties. The five experimental salinities were divided into one region, and SOD activity was the highest in this region. In a comprehensive analysis of stability and activity, SOD had the best activity and stability. The results of AMMI analysis for temperature reveal that genotype, temperature, and genotype × temperature interaction had significant effects on the antioxidant properties (p < 0.001); 82.4720%, 4.0666%, and 12.0968% of the total sum of squares for antioxidant property activity were attributable to the effects of genotype, temperature, and genotype × temperature interaction, respectively. The results of GGE biplot analysis for temperature reveal a large difference in the activity ranking of antioxidant properties between 17 °C and the other four temperatures, while only small differences in the activity rankings were detected among the other four temperatures. The difference in the activity ranking of antioxidant properties was greatest between the temperatures of 17 and 20 °C. A temperature of 17 °C showed the strongest ability to distinguish the three antioxidant properties. Additionally, the five test temperatures were grouped into one region, and comprehensive analysis of activity and stability showed that SOD had the best activity and stability.

An explainable machine learning method for assessing surgical skill in liposuction surgery.

PURPOSE: Surgical skill assessment has received growing interest in surgery training and quality control due to its essential role in competency assessment and trainee feedback. However, the current assessment methods rarely provide corresponding feedback guidance while giving ability evaluation. We aim to validate an explainable surgical skill assessment method that automatically evaluates the trainee performance of liposuction surgery and provides visual postoperative and real-time feedback. METHODS: In this study, machine learning using a model-agnostic interpretable method based on stroke segmentation was introduced to objectively evaluate surgical skills. We evaluated the method on liposuction surgery datasets that consisted of motion and force data for classification tasks. RESULTS: Our classifier achieved optimistic accuracy in clinical and imitation liposuction surgery models, ranging from 89 to 94%. With the help of SHapley Additive exPlanations (SHAP), we deeply explore the potential rules of liposuction operation between surgeons with variant experiences and provide real-time feedback based on the ML model to surgeons with undesirable skills. CONCLUSION: Our results demonstrate the strong abilities of explainable machine learning methods in objective surgical skill assessment. We believe that the machine learning model based on interpretive methods proposed in this article can improve the evaluation and training of liposuction surgery and provide objective assessment and training guidance for other surgeries.

Three-Dimensional Quantitative Coherent Diffraction Imaging of Staphylococcus aureus Treated with Peptide-Mineralized Au-Cluster Probes.

Characterizing interactions between microbial cells and their specific inhibitory drugs is essential for developing effective drugs and understanding the therapeutic mechanism. Functional metal nanoclusters can be effective inhibitory agents against microorganisms according to various characterization methods, but quantitative three-dimensional (3D) spatial structural analysis of intact cells is lacking. Herein, using coherent X-ray diffraction imaging, we performed in situ 3D visualization of unstained Staphylococcus aureus cells treated with peptide-mineralized Au-cluster probes at a resolution of ∼47 nm. Subsequent 3D mass-density mapping and quantitative structural analyses of S. aureus in different degrees of destruction showed that the bacterial cell wall was damaged and cytoplasmic constituents were released from cells, confirming the significant antibacterial effects of the Au-cluster probe. This study provides a promising nondestructive approach for quantitative imaging and paves the way for further research into microbe-inhibitor drug interactions.

Structural and functional characterization of turbot pparγ: Activation during high temperature and regulation of lipid metabolism.

Transcription factors of the peroxisome proliferator-activated receptor gamma (pparγ) is a ligand-activated receptor that plays key roles in lipid metabolism and inflammation. In this study, we cloned the pparγ cDNA of turbot (Scophthalmus maximus). It has a 1659 bp coding sequence (CDS) and encodes 552 amino acids. The deduced PPARγ protein of turbot contains two highly conserved domains, a C4-type zinc finger, a nuclear hormone receptor DNA-binding region signature, and a HOLI domain (ligand-binding domain of hormone receptors) identical to that of in other species. qPCR results showed that the expression level of pparγ and the expression of the fatty acid transporters fatp and cd36 were significantly increased under high-temperature stress. This indicates that high temperatures activate the transcriptional activity of pparγ, and lipid metabolism plays an important role in turbot under high-temperature stress. In addition, RNA interference was used to explore the regulation of pparγ on lipid metabolism of turbot at high temperatures. The results showed that the mRNA level of pparγ was significantly decreased. After the expression level of pparγ was inhibited, the expression levels of fatp and cd36 were significantly decreased. At the same time, GW9662 (a pparγ antagonist) was used to inhibit pparγ activity in turbot kidney cells, and fatp and cd36 gene expressions were detected. The mRNA expression levels of pparγ, fatp, and cd36 were significantly decreased. Our results suggest that high temperatures activate pparγ in turbot and that pparγ regulates lipid transport and maintains lipid metabolism homeostasis through positive regulation of the expression of downstream genes fatp and cd36. This study further elucidates that the pparγ-mediated signaling pathway may play an important role in regulating lipid metabolism during heat stress in teleost fish.

Dissecting the Molecular Regulation of Natural Variation in Growth and Senescence of Two Eutrema salsugineum Ecotypes.

Salt cress (Eutrema salsugineum, aka Thellungiella salsuginea) is an extremophile and a close relative of Arabidopsis thaliana. To understand the mechanism of selection of complex traits under natural variation, we analyzed the physiological and proteomic differences between Shandong (SD) and Xinjiang (XJ) ecotypes. The SD ecotype has dark green leaves, short and flat leaves, and more conspicuous taproots, and the XJ ecotype had greater biomass and showed clear signs of senescence or leaf shedding with age. After 2-DE separation and ESI-MS/MS identification, between 25 and 28 differentially expressed protein spots were identified in shoots and roots, respectively. The proteins identified in shoots are mainly involved in cellular metabolic processes, stress responses, responses to abiotic stimuli, and aging responses, while those identified in roots are mainly involved in small-molecule metabolic processes, oxidation-reduction processes, and responses to abiotic stimuli. Our data revealed the evolutionary differences at the protein level between these two ecotypes. Namely, in the evolution of salt tolerance, the SD ecotype highly expressed some stress-related proteins to structurally adapt to the high salt environment in the Yellow River Delta, whereas the XJ ecotype utilizes the specialized energy metabolism to support this evolution of the short-lived xerophytes in the Xinjiang region.

The oral microbiome analysis reveals the similarities and differences between periodontitis and Crohn's disease-associated periodontitis.

Patients with Crohn's disease (CD) have higher incidences of oral diseases such as dental caries and periodontitis than healthy people. Studies indicate that the interaction between gut and oral microbiota is an important factor. To compare the composition and diversity of the oral microbiome in periodontitis and CD-associated periodontitis, subgingival plaque and saliva samples from patients with these diseases were collected for 16S rRNA gene sequencing analyses. In CD-associated periodontitis, the subgingival plaque had greater microbial diversity than saliva. Subgingival plaque had decreased abundances of Firmicutes, Streptococcus, and Haemophilus and increased abundances of Bacteroidetes, Actinomyces, Treponema_2, Capnocytophaga, and Porphyromonas relative to saliva. The microbial composition in subgingival plaque was similar between the two diseases. Both red complex (Porphyromonas, Tannerella, and Treponema) and orange complex (Fusobacteria) bacteria were abundant in periodontitis subgingival plaque, while orange complex bacteria (Prevotella_2 and Prevotella) were abundant in CD-associated periodontitis subgingival plaque. Pocket depth was significantly positively correlated with multiple periodontal pathogens, including Porphyromonas, Tannerella, and Treponema. This study reveals the similarities and differences in the oral microbiome between periodontitis and CD-associated periodontitis, which provides a foundation to further explore the associations between CD and periodontitis.

Dual role of CsrA in regulating the hemolytic activity of Escherichia coli O157:H7.

Post-transcriptional global carbon storage regulator A (CsrA) is a sequence-specific RNA-binding protein involved in the regulation of multiple bacterial processes. Hemolysin is an important virulence factor in the enterohemorrhagic Escherichia coli O157:H7 (EHEC). Here, we show that CsrA plays a dual role in the regulation of hemolysis in EHEC. CsrA significantly represses plasmid-borne enterohemolysin (EhxA)-mediated hemolysis and activates chromosome-borne hemolysin E (HlyE)-mediated hemolysis through different mechanisms. RNA structure prediction revealed a well-matched stem-loop structure with two potential CsrA binding sites located on the 5' untranslated region (UTR) of ehxB, which encodes a translocator required for EhxA secretion. CsrA inhibits EhxA secretion by directly binding to the RNA leader sequence of ehxB to repress its expression in two different ways: CsrA either binds to the Shine-Dalgarno sequence of ehxB to block ribosome access or to ehxB transcript to promote its mRNA decay. The predicted CsrA-binding site 1 of ehxB is essential for its regulation. There is a single potential CsrA-binding site at the 5'-end of the hlyE transcript, and its mutation completely abolishes CsrA-dependent activation. CsrA can also stabilize hlyE mRNA by directly binding to its 5' UTR. Overall, our results indicate that CsrA acts as a hemolysis modulator to regulate pathogenicity under certain conditions.

Quantitative analysis of the effect of radiation on mitochondria structure using coherent diffraction imaging with a clustering algorithm.

Radiation damage and a low signal-to-noise ratio are the primary factors that limit spatial resolution in coherent diffraction imaging (CDI) of biomaterials using X-ray sources. Introduced here is a clustering algorithm named ConvRe based on deep learning, and it is applied to obtain accurate and consistent image reconstruction from noisy diffraction patterns of weakly scattering biomaterials. To investigate the impact of X-ray radiation on soft biomaterials, CDI experiments were performed on mitochondria from human embryonic kidney cells using synchrotron radiation. Benefiting from the new algorithm, structural changes in the mitochondria induced by X-ray radiation damage were quantitatively characterized and analysed at the nanoscale with different radiation doses. This work also provides a promising approach for improving the imaging quality of biomaterials with XFEL-based plane-wave CDI.

Ultrafast Single-Particle Imaging with Intense X-Ray Pulses.

Ultrafast single-particle imaging with intense x-ray pulses from free-electron laser sources provides a new approach for visualizing structure and dynamics on the nanoscale. After a short introduction to the novel free-electron laser sources and methods, we highlight selected applications and discuss how ultrafast imaging flourishes from method development to early applications in physics and biology to opportunities for chemical sciences.

Transcriptome analysis reveals that high temperatures alter modes of lipid metabolism in juvenile turbot (Scophthalmus maximus) liver.

With the increase in farming density and the continuously high summer temperatures against the background of global warming, high temperature stress has become a major challenge in fish farming. In this study, we simulated the high temperature environments (20 °C, 24 °C, and 28 °C) that may occur during turbot culture. High-throughput sequencing was used to investigate the lipid metabolism response patterns in juvenile turbot liver under high temperature stress. A total of 2067 differentially expressed genes (DEGs) were identified. KEGG analysis revealed that the DEGs were mainly associated with glycerophospholipid metabolism, steroid biosynthesis, glycerolipid metabolism, fatty acid metabolic pathways, and the PPAR signaling pathway. A regulatory network was constructed to further elucidate the transcriptional regulation of lipid metabolism. We speculated that high temperature activates PPAR signaling pathway through interaction with ligands such as fatty acids. On the one hand, the HMGCS1 gene in this pathway can inhibit sterol synthesis by down-regulating the expression of key genes in steroid biosynthesis pathway (SQLE, EBP, and DHCR24). On the other hand, the expression of ACSL1 in this pathway is significantly increased under high temperature, which may play an important role in regulating fatty acid metabolism. Moreover, we collected blood and detected changes in serum lipid parameters; the variation patterns were also consistent with our results. These findings reveal that lipid metabolism has an important regulatory role in stress resistance when turbot is exposed to high temperatures.

Genetic parameters of seven immune factors in turbot (Scophthalmus maximus) infected with Vibrio anguillarum.

In this study, 1,800 turbot (Scophthalmus maximus) individuals from 30 full-sib families were experimentally infected with Vibrio anguillarum, and the expression levels of the immune factors lysozyme, hepcidin, heat-shock protein 70 (HSP70), HSP90, immunoglobulin M (IgM), C-type lectin and Lily-type lectin in the liver were measured by real-time PCR. Heritability values of the seven immune factors were 0.289 ± 0.087, 0.092 ± 0.024, 0.282 ± 0.043, 0.244 ± 0.027, 0.343 ± 0.081, 0.092 ± 0.011 and 0.084 ± 0.009, respectively. The ranges of phenotypic, genetic and environmental correlations were -0.889 to 0.759, -0.841 to 0.888 and -0.919 to 0.883, respectively. The heritability values of HSP70, HSP90 and IgM were moderate, and the genetic correlations between HSP70, HSP90 and IgM were moderate to highly positive, which suggests that the immunocompetence of turbot against V. anguillarum can be improved by genetically improving these three immune characters via multi-trait integrated breeding technology or indirect selection.

Diffraction data from aerosolized Coliphage PR772 virus particles imaged with the Linac Coherent Light Source.

Single Particle Imaging (SPI) with intense coherent X-ray pulses from X-ray free-electron lasers (XFELs) has the potential to produce molecular structures without the need for crystallization or freezing. Here we present a dataset of 285,944 diffraction patterns from aerosolized Coliphage PR772 virus particles injected into the femtosecond X-ray pulses of the Linac Coherent Light Source (LCLS). Additional exposures with background information are also deposited. The diffraction data were collected at the Atomic, Molecular and Optical Science Instrument (AMO) of the LCLS in 4 experimental beam times during a period of four years. The photon energy was either 1.2 or 1.7 keV and the pulse energy was between 2 and 4 mJ in a focal spot of about 1.3 µm x 1.7 µm full width at half maximum (FWHM). The X-ray laser pulses captured the particles in random orientations. The data offer insight into aerosolised virus particles in the gas phase, contain information relevant to improving experimental parameters, and provide a basis for developing algorithms for image analysis and reconstruction.

Generation and simple characterization of flat, liquid jets.

We present an approach to determine the absolute thickness profile of flat liquid jets, which takes advantage of the information of thin film interference combined with light absorption, both captured in a single microscopic image. The feasibility of the proposed method is demonstrated on our compact experimental setup used to generate micrometer thin, free-flowing liquid jet sheets upon collision of two identical laminar cylindrical jets. Stable operation was achieved over several hours of the flat jet in vacuum (10-4 mbar), making the system ideally suitable for soft x-ray photon spectroscopy of liquid solutions. We characterize the flat jet size and thickness generated with two solvents, water and ethanol, employing different flow rates and nozzles of variable sizes. Our results show that a gradient of thickness ranging from a minimal thickness of 2 µm to over 10 µm can be found within the jet surface area. This enables the tunability of the sample thickness in situ, allowing the optimization of the transmitted photon flux for the chosen photon energy and sample. We demonstrate the feasibility of x-ray absorption spectroscopy experiments in transmission mode by measuring at the oxygen K-edge of ethanol. Our characterization method and the description of the experimental setup and its reported performance are expected to expand the range of applications and facilitate the use of flat liquid jets for spectroscopy experiments.

Impacts of geographic factors and population density on the COVID-19 spreading under the lockdown policies of China.

The outbreak of COVID-19 pandemic has a high spreading rate and a high fatality rate. To control the rapid spreading of COVID-19 virus, Chinese government ordered lockdown policies since late January 2020. The aims of this study are to quantify the relationship between geographic information (i.e., latitude, longitude and altitude) and cumulative infected population, and to unveil the importance of the population density in the spreading speed during the lockdown. COVID-19 data during the period from December 8, 2019 to April 8, 2020 were collected before and after lockdown. After discovering two important geographic factors (i.e., latitude and altitude) by estimating the correlation coefficients between each of them and cumulative infected population, two linear models of cumulative infected population and COVID-19 spreading speed were constructed based on these two factors. Overall, our findings from the models showed a negative correlation between the provincial daily cumulative COVID-19 infected number and latitude/altitude. In addition, population density is not an important factor in COVID-19 spreading under strict lockdown policies. Our study suggests that lockdown policies of China can effectively restrict COVID-19 spreading speed.