Identification of PANoptosis-related subtypes, construction of a prognosis signature, and tumor microenvironment landscape of hepatocellular carcinoma using bioinformatic analysis and experimental verification.

Background: Hepatocellular carcinoma (HCC) is one of the most lethal malignancies worldwide. PANoptosis is a recently unveiled programmed cell death pathway, Nonetheless, the precise implications of PANoptosis within the context of HCC remain incompletely elucidated. Methods: We conducted a comprehensive bioinformatics analysis to evaluate both the expression and mutation patterns of PANoptosis-related genes (PRGs). We categorized HCC into two clusters and identified differentially expressed PANoptosis-related genes (DEPRGs). Next, a PANoptosis risk model was constructed using LASSO and multivariate Cox regression analyses. The relationship between PRGs, risk genes, the risk model, and the immune microenvironment was studies. In addition, drug sensitivity between high- and low-risk groups was examined. The expression profiles of these four risk genes were elucidate by qRT-PCR or immunohistochemical (IHC). Furthermore, the effect of CTSC knock down on HCC cell behavior was verified using in vitro experiments. Results: We constructed a prognostic signature of four DEPRGs (CTSC, CDCA8, G6PD, and CXCL9). Receiver operating characteristic curve analyses underscored the superior prognostic capacity of this signature in assessing the outcomes of HCC patients. Subsequently, patients were stratified based on their risk scores, which revealed that the low-risk group had better prognosis than those in the high-risk group. High-risk group displayed a lower Stromal Score, Immune Score, ESTIMATE score, and higher cancer stem cell content, tumor mutation burden (TMB) values. Furthermore, a correlation was noted between the risk model and the sensitivity to 56 chemotherapeutic agents, as well as immunotherapy efficacy, in patient with. These findings provide valuable guidance for personalized clinical treatment strategies. The qRT-PCR analysis revealed that upregulated expression of CTSC, CDCA8, and G6PD, whereas downregulated expression of CXCL9 in HCC compared with adjacent tumor tissue and normal liver cell lines. The knockdown of CTSC significantly reduced both HCC cell proliferation and migration. Conclusion: Our study underscores the promise of PANoptosis-based molecular clustering and prognostic signatures in predicting patient survival and discerning the intricacies of the tumor microenvironment within the context of HCC. These insights hold the potential to advance our comprehension of the therapeutic contribution of PANoptosis plays in HCC and pave the way for generating more efficacious treatment strategies.

Asymmetry-enhanced attention network for Alzheimer's diagnosis with structural Magnetic Resonance Imaging.

BACKGROUND AND OBJECTIVE: With the aging of the global population becoming severe, Alzheimer's disease (AD) has become one of the world's most common senile diseases. Many studies have suggested that the brain's left-right asymmetry is one of the possible diagnostic landmarks for AD. However, most published approaches to classification problems may not adequately explore the asymmetry between the left and right hemispheres. At the same time, the relationship between asymmetry traits and other classifier features remains understudied. METHODS: In this paper, we proposed an asymmetry enhanced attention network (ASEAN) for AD diagnosis that effectively combines the anatomical asymmetry characteristics of the brain to enhance the accuracy and stability of classification tasks. First, we proposed a multi-scale asymmetry feature extraction module (MSAF) that can extract the asymmetry features of the brain from various scales. Second, we proposed an asymmetry refinement module (ARM) that considers the dependency between feature maps to suppress the irrelevant regions of the asymmetric feature maps. In addition, a parameter-free attention module was introduced to infer 4D attention weights and improve the network's representation capability. RESULTS: The proposed method achieved performance improvements on two databases: Alzheimer's Disease Neuroimaging Initiative (ADNI) and Australian Imaging, Biomarkers and Lifestyle (AIBL). For the classification tasks on ADNI, the proposed method achieves 92.1% accuracy, 96.2% sensitivity, and 91.3% specificity on the AD vs. CN (Cognitively Normal) task. Compared with state-of-the-art methods, the proposed method could achieve comparable results. CONCLUSION: The proposed model can extract long-range left-right brain similarity as complementary information and improve the model's diagnostic performance. A large number of experiments also support the model's validity. At the same time, this work provides a valuable reference for other neurological diseases, particularly those that exhibit left-right brain asymmetry during development.

EA-Net: Edge-aware network for brain structure segmentation via decoupled high and low frequency features.

Automatic brain structure segmentation in Magnetic Resonance Image (MRI) plays an important role in the diagnosis of various neuropsychiatric diseases. However, most existing methods yield unsatisfactory results due to blurred boundaries and complex structures. Improving the segmentation ability requires the model to be explicit about the spatial localization and shape appearance of targets, which correspond to the low-frequency content features and the high-frequency edge features, respectively. Therefore, in this paper, to extract rich edge and content feature representations, we focus on the composition of the feature and utilize a frequency decoupling (FD) block to separate the low-frequency and high-frequency parts of the feature. Further, a novel edge-aware network (EA-Net) is proposed for jointly learning to segment brain structures and detect object edges. First, an encoder-decoder sub-network is utilized to obtain multi-level information from the input MRI, which is then sent to the FD block to complete the frequency separation. Further, we use different mechanisms to optimize both the low-frequency and high-frequency features. Finally, these two parts are fused to generate the final prediction. In particular, we extract the content mask and the edge mask from the optimization feature with different supervisions, which forces the network to learn the boundary features of the object. Extensive experiments are performed on two public brain MRI T1 scan datasets (the IBSR dataset and the MALC dataset) to evaluate the effectiveness of the proposed algorithm. The experiments show that the EA-Net achieves the best performance compared with the state-of-the-art methods, and improves the segmentation DSC score by 1.31% at most compared with the U-Net model and its variants. Moreover, we evaluate the EA-Net under different noise disturbances, and the results demonstrate the robustness and superiority of our method under low-quality noisy MRI. Code is available at https://github.com/huqian999/EA-Net.

Rapid generation of maternal mutants via oocyte transgenic expression of CRISPR-Cas9 and sgRNAs in zebrafish.

Maternal products are exclusive factors to drive oogenesis and early embryonic development. As disrupting maternal gene functions is either time-consuming or technically challenging, early developmental programs regulated by maternal factors remain mostly elusive. We provide a transgenic approach to inactivate maternal genes in zebrafish primary oocytes. By introducing three tandem single guide RNA (sgRNA) expression cassettes and a green fluorescent protein (GFP) reporter into Tg(zpc:zcas9) embryos, we efficiently obtained maternal nanog and ctnnb2 mutants among GFP-positive F1 offspring. Notably, most of these maternal mutants displayed either sgRNA site-spanning genomic deletions or unintended large deletions extending distantly from the sgRNA targets, suggesting a prominent deletion-prone tendency of genome editing in the oocyte. Thus, our method allows maternal gene knockout in the absence of viable and fertile homozygous mutant adults. This approach is particularly time-saving and can be applied for functional screening of maternal factors and generating genomic deletions in zebrafish.

A Time-Saving Strategy to Generate Double Maternal Mutants by an Oocyte-Specific Conditional Knockout System in Zebrafish.

Maternal products are those mRNAs and proteins deposited during oogenesis, which play critical roles in controlling oocyte formation, fertilization, and early embryonic development. However, loss-of-function studies for these maternal factors are still lacking, mainly because of the prolonged period of transgenerational screening and technical barriers that prevent the generation of maternal (M) and maternal and zygotic (MZ) mutant embryos. By the transgenic expression of multiple sgRNAs targeting a single gene of interest in the background of a transgenic line Tg(zpc:zcas9) with oocyte-specific cas9 expression, we have successfully obtained maternal or maternal-zygotic mutant for single genes in F1 embryos. In this work, we tandemly connected a maternal GFP marker and eight sgRNA expression units to target dvl2 and dvl3a simultaneously and introduced this construct to the genome of Tg(zpc:zcas9) by meganuclease I-Sce I. As expected, we confirmed the existence of Mdvl2;Mdvl3a embryos with strong defective convergence and extension movement during gastrulation among outcrossed GFP positive F1 offspring. The MZdvl2;MZdvl3a embryos were also obtained by crossing the mutant carrying mosaic F0 female with dvl2+/-;dvl3a-/- male fish. This proof-of-principle thus highlights the potential of this conditional knockout strategy to circumvent the current difficulty in the study of genes with multiple functionally redundant paralogs.

Visualisation of flooding along an unvegetated, ephemeral river using Google Earth Engine: Implications for assessment of channel-floodplain dynamics in a time of rapid environmental change.

Given rapid environmental change, the development of new, data-driven, interdisciplinary approaches is essential for improving assessment and management of river systems, especially with respect to flooding. In the world's extensive drylands, difficulties in obtaining field observations of major hydrological events mean that remote sensing techniques are commonly used to map river floods and assess flood impacts. Such techniques, however, are dependent on available cloud-free imagery during or immediately after peak discharge, and single images may omit important flood-related hydrogeomorphological events. Here, we combine multiple Landsat images from Google Earth Engine (GEE) with precipitation datasets and high-resolution (<0.65 m) satellite imagery to visualise flooding and assess the associated channel-floodplain dynamics along a 25 km reach of the unvegetated, ephemeral Río Colorado, Bolivia. After cloud and shadow removal, Landsat surface reflectance data were used to calculate the Modified Normalized Difference Water Index (MNDWI) and map flood extents and patterns. From 2004 through 2016, annual flooding area along the narrow (<30 m), shallow (<1.7 m), fine-grained (dominantly silt/clay) channels was positively correlated (R2 = 0.83) with 2-day maximum precipitation totals. Rapid meander bend migration, bank erosion, and frequent overbank flooding was associated with formation of crevasse channels, splays, and headward-eroding channels, and with avulsion (shifting of flow from one channel to another). These processes demonstrate ongoing, widespread channel-floodplain dynamics despite low stream powers and cohesive sediments. Application of our study approaches to other dryland rivers will help generate comparative data on the controls, rates, patterns and timescales of channel-floodplain dynamics under scenarios of climate change and direct human impacts, with potential implications for improved river management.

[Usage and efficacy of timolol maleate eye drops in treatment of superficial infantile hemangioma].

OBJECTIVE: To determine drug dose and usage of timolol maleate eye drops in the treatment of superficial infantile hemangioma.
 Methods: A total of 250 superficial hemangioma infants were recruited and assigned into 5 groups (n=50 for each group): an external application group and 4 exterior coating groups (2, 4, 6, 8 times per day). We evaluated the therapeutic effect of different methods for drug application (external application or exterior coating) and the frequency for drug administration on superficial infantile hemangioma.
 Results: The external application group (twice a day and 0.5 hour per time) showed better effect than that in the exterior coating group with twice a day (P<0.001). The difference in therapeutic effect between the exterior coating group with 6 times a day and exterior coating group with twice a day or with 3 times a day was significant (P<0.001). The differences in drug efficacy were not found among the exterior coating group with 6 times a day, the exterior coating group with 8 times a day, or the external application group with twice a day (All P>0.05).
 Conclusion: Drug dose may affect the therapeutic effect of timolol maleate eye drops in superficial hemangioma infants, and exterior coating with 6 times a day may achieve the best curative effect.

[Endovascular treatment for venous malformations with three-point anhydrous ethanol injection].

OBJECTIVE: To explore the safe method with anhydrous ethanol injection in the treatment of venous malformation.
 METHODS: A total of 96 patients with venous malformation were conducted anhydrous ethanol injection for 245 times through percutaneous puncture by three-point method. The complications were observed. In animal experiment, according to the different concentrations of anhydrous ethanol injection, rats were divided into an anhydrous ethanol group, a 75% ethanol group, a 50% ethanol group and a 25% ethanol group (n=5 in each group), and the damage of vessels after ethanol injection was observed.
 RESULTS: The successful rate for three-point ethanol injection was 88%. The incidence for both skin ulcer and numbness was 0.9% without severe complications in lung and heart. In the animal experiments, the entire vessel wall including outer membrane was damaged in the anhydrous ethanol group. Part of vessel walls, including the inner membrane and muscle layer, were damaged in both the 75% ethanol group and the 50% ethanol group. However, there was no damage in the vessels in the 25% ethanol group.
 CONCLUSION: With the decrease in ethanol concentration, the vascular damage is decreased and eventually disappeared. Three-point anhydrous ethanol injection is safe and effective.

MacroH2A suppresses the proliferation of the B16 melanoma cell line.

MacroH2A is the most frequently altered histone, which participates in cancer progression. Increasing evidence demonstrates that cancer progression could be regulated by macroH2A by affecting the cell cycle. In the present study, it was demonstrated that macroH2A suppresses melanoma cell progression and the molecular mechanisms underlying this process were examined. The interference and overexpression vectors of macroH2A were constructed and then transferred into B16 melanoma cells and, following transfection, were analyzed by quantitative polymerase chain reaction (PCR), western blot analysis and immunofluorescence assays. Apoptosis and the cell cycle stage among all the treatment groups were detected. Then, cyclin D1, cyclin D3, cyclin-dependent protein kinase (CDK) 4, CDK6 and CDK8 expression was detected in order to elucidate the effects of macroH2A on cell cycle-related genes. The results demonstrated that the overexpression of macroH2A suppressed melanoma cell progression and arrested the cells in the G2/M stage. Furthermore, macroH2A inhibits cyclin D1, cyclin D2, CDK6 and CDK8 expression in B16 melanoma cells. In conclusion, the results demonstrated that macroH2A, a critical component of chromatin, suppresses the development of melanoma (which results from a disordered cell cycle) through regulating cyclin D1, cyclin D3 and CDK6 genes.