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Purpose: This study aimed to evaluate a radiomics model using Photoacoustic/ultrasound (PA/US) imaging at intra and peri-tumoral area to differentiate Luminal and non-Luminal breast cancer (BC) and to determine the optimal peritumoral area for accurate classification. Materials and methods: From February 2022 to April 2024, this study continuously collected 322 patients at Shenzhen People’s Hospital, using standardized conditions for PA/US imaging of BC. Regions of interest were delineated using ITK-SNAP, with peritumoral regions of 2 mm, 4 mm, and 6 mm automatically expanded using code from the Pyradiomic package. Feature extraction was subsequently performed using Pyradiomics. The study employed Z-score normalization, Spearman correlation for feature correlation, and LASSO regression for feature selection, validated through 10-fold cross-validation. The radiomics model integrated intra and peri-tumoral area, evaluated by receiver operating characteristic curve(ROC), Calibration and Decision Curve Analysis(DCA). Results: We extracted and selected features from intratumoral and peritumoral PA/US images regions at 2 mm, 4 mm, and 6 mm. The comprehensive radiomics model, integrating these regions, demonstrated enhanced diagnostic performance, especially the 4 mm model which showed the highest area under the curve(AUC):0.898(0.78–1.00) and comparably high accuracy (0.900) and sensitivity (0.937). This model outperformed the standalone clinical model and combined clinical-radiomics model in distinguishing between Luminal and non-Luminal BC, as evidenced in the test set results. Conclusion: This study developed a radiomics model integrating intratumoral and peritumoral at 4 mm region PA/US model, enhancing the differentiation of Luminal from non-Luminal BC. It demonstrated the diagnostic utility of peritumoral characteristics, reducing the need for invasive biopsies and aiding chemotherapy planning, while emphasizing the importance of optimizing tumor surrounding size for improved model accuracy.

The CCAAT enhancer binding protein alpha (CEBPA) is crucial for myeloid differentiation and the balance of haematopoietic stem and progenitor cell (HSPC) quiescence and self-renewal, and its dysfunction can drive leukemogenesis. However, its role in HSPC generation has not been fully elucidated. Here, we utilized various zebrafish cebpa mutants to investigate the function of Cebpa in the HSPC compartment. Co-localization analysis showed that cebpa expression is enriched in nascent HSPCs. Complete loss of Cebpa function resulted in a significant reduction in early HSPC generation and the overall HSPC pool during embryonic haematopoiesis. Interestingly, while myeloid differentiation was impaired in cebpa N-terminal mutants expressing the truncated zP30 protein, the number of HSPCs was not affected, indicating a redundant role of Cebpa P42 and P30 isoforms in HSPC development. Additionally, epistasis experiments confirmed that Cebpa functions downstream of Runx1 to regulate HSPC emergence. Our findings uncover a novel role of Cebpa isoforms in HSPC generation and maintenance, and provide new insights into HSPC development.

Purpose To assess whether gray-scale ultrasound (US) based radiomic features can help distinguish HER2 expressions (ie, HER2-overexpressing, HER2-low-expressing, and HER2-zero-expressing) in breast cancer. Materials and Methods This retrospective study encompassed female breast cancer patients who underwent US examinations at two distinct centers from February 2021 to July 2023. Tumor segmentation and radiomic feature extraction were performed on grayscale US images. Decision Tree analysis was employed to simultaneously evaluate feature importance, and the Least Absolute Shrinkage and Selection Operator technique was utilized for feature selection to construct the radiomic signature. The Area Under the Curve (AUC) of the Receiver Operating Characteristic curve was employed to assess the performance of the radiomic features. Multivariate logistic regression was used to identify independent predictors for distinguishing HER2 expression in the dataset. Results The training set comprised 292 patients from Center 1 (median, 51 years; interquartile range [IQR]: 45-61), while the external validation set included 131 patients from Center 2 (median, 51 years; IQR: 45-62). In the external validation dataset, the radiomic features achieved AUC of 0.76 for distinguishing between HER2-low and positive tumors versus HER2-zero tumors. The AUC for differentiating HER2-low (1+) from HER2-zero tumors was 0.74, and for distinguishing HER2-low (2+) from HER2-zero tumors, the AUC was 0.77. In the multivariate analysis assessing HER2-low and HER2-positive versus HER2-zero tumors, internal echoes (P = .029) and margins (P

Abstract Background Continuous peripheral nerve blocks are widely used for anesthesia and postoperative analgesia in lower limb surgeries. The authors aimed to develop a novel continuous sacral plexus block procedure for analgesia during total knee arthroplasty. Methods The study comprised two stages. In Stage I, the authors built upon previous theories and technological innovations to develop a novel continuous sacral plexus block method, ultrasound-guided continuous parasacral ischial plane block (UGCPIPB) and subsequently conducted a proof-of-concept study to assess its effectiveness and feasibility. Stage II involved a historical control study to compare clinical outcomes between patients undergoing this new procedure and those receiving the conventional procedure. Results The study observed a 90% success rate in catheter placement. On postoperative day (POD) 1, POD2, and POD3, the median visual analog scale (VAS) scores were 3 (range, 1.5–3.5), 2.5 (1.6–3.2), and 2.7 (1.3–3.4), respectively. Furthermore, 96.3% of the catheters remained in place until POD3, as confirmed by ultrasound. The study revealed a significant increase in skin temperature and peak systolic velocity of the anterior tibial artery on the blocked side compared with those on the non-blocked side. Complications included catheter clogging in one patient and leakage at the insertion site in two patients. In Stage II, the novel technique was found to be more successful than conventional techniques, with a lower catheter displacement rate than the conventional procedure for continuous sciatic nerve block. Conclusion UGCPIPB proved to be an effective procedure and safe for analgesia in total knee arthroplasty. Chinese Clinical Trial Registry Number ChiCTR2300068902.

Abstract Background Despite the success of tyrosine kinase inhibitors in chronic myeloid leukemia (CML) therapy, CML still faces the challenges of drug resistance and progression to blast crisis. Twenty‐five percent of patients have imatinib resistance and treatment difficulties due to heterogeneity after progression, but little is known about the mechanism. A key transcription factor in hematopoiesis, MYB, has been reported to increase abnormally in several types of aggressive blood disorders including CML. Methods This study used a zebrafish model to explore the relationship between BCR/ABL1 and c‐myb in CML progression. A CML zebrafish model was crossed with a c‐myb hyperactivity transgenic line. Results It was found that both exogenous BCR/ABL1 and c‐myb could up‐regulate the expression of neutrophil‐related genes. More seriously, neutrophil accumulation was observed when BCR/ABL1 was combined with c‐myb overexpression. Further studies showed that c‐myb may be one of the downstream targets of BCR/ABL1 and the effect of BCR/ABL1 on neutrophils was c‐myb dependent. Taking advantage of this inheritable in vivo model, it was shown that a combination of imatinib and flavopiridol, a cyclin‐dependent kinase inhibitor targeting MYB, could more effectively alleviate the aggressive phenotype of the double transgene line. Conclusion In summary, this study suggests that c‐myb acts downstream of BCR/ABL1 and is involved in CML progression and is therefore a risk factor and a valuable target for the treatment of CML progression. The model used in the study could be helpful in high‐throughput drug screening in CML transformation.

Abstract During the development of the vertebrate nervous system, 50% of the nerve cells undergo apoptosis shortly after formation. This process is important for sculpting tissue during morphogenesis and removing transiently functional cells that are no longer needed, ensuring the appropriate number of neurons in each region. Dysregulation of neuronal apoptosis can lead to neurodegenerative diseases. However, the molecular events involved in activating and regulating the neuronal apoptosis program are not fully understood. In this study, we identified several RB1 mutations in patients with neurodegenerative diseases. Then, we used a zebrafish model to investigate the role of Rb1 in neuronal apoptosis. We showed that Rb1-deficient mutants exhibit a significant hindbrain neuronal apoptosis, resulting in increased microglia infiltration. We further revealed that the apoptotic neurons in Rb1-deficient zebrafish were post-mitotic neurons, and Rb1 inhibits the apoptosis of these neurons by regulating bcl2/caspase through binding to Kmt5b. Moreover, using this zebrafish mutant, we verified the pathogenicity of the R621S and L819V mutations of human RB1 in neuronal apoptosis. Collectively, our data indicate that the Rb1-Kmt5b-caspase/bcl2 axis is crucial for protecting post-mitotic neurons from apoptosis and provides an explanation for the pathogenesis of clinically relevant mutations.

Abstract An ideal approach for the construction of aryl boron compounds is to selectively replace a C–H bond in arenes with a C–B bond, and controlling regioselectivity is one of the most challenging aspects of these transformations. Herein, we report an iridium-catalyzed trialkoxysilane protecting group-assisted regioselective C–H borylation of arenes, including derivatives of benzaldehydes, acetophenones, benzoic acids, benzyl alcohols, phenols, aryl silanes, benzyl silanes, and multi-functionalized aromatic rings are all well tolerated and gave the para -selective C–H borylation products in a short time without the requirement of inert gases atmosphere. The site-selective C–H borylation can be adjustable by installing the developed trialkoxysilane protecting group on different functional groups on one aromatic ring. Importantly, the preparation process of the trialkoxychlorosilane is efficient and scalable. Mechanistic and computational studies reveal that the steric hindrance of the trialkoxysilane protecting group plays a key role in dictating the para-selectivity.

Background: The differentiation between benign and malignant breast tumors extends beyond morphological structures to encompass functional alterations within the nodules. The combination of photoacoustic (PA) imaging and radiomics unveils functional insights and intricate details that are imperceptible to the naked eye. Purpose: This study aims to assess the efficacy of PA imaging in breast cancer radiomics, focusing on the impact of peritumoral region size on radiomic model accuracy. Materials and methods: From January 2022 to November 2023, data were collected from 358 patients with breast nodules, diagnosed via PA/US examination and classified as BI-RADS 3–5. The study used the largest lesion dimension in PA images to define the region of interest, expanded by 2 mm, 5 mm, and 8 mm, for extracting radiomic features. Techniques from statistics and machine learning were applied for feature selection, and logistic regression classifiers were used to build radiomic models. These models integrated both intratumoral and peritumoral data, with logistic regressions identifying key predictive features. Results: The developed nomogram, combining 5 mm peritumoral data with intratumoral and clinical features, showed superior diagnostic performance, achieving an AUC of 0.950 in the training cohort and 0.899 in validation. This model outperformed those based solely on clinical features or other radiomic methods, with the 5 mm peritumoral region proving most effective in identifying malignant nodules. Conclusion: This research demonstrates the significant potential of PA imaging in breast cancer radiomics, especially the advantage of integrating 5 mm peritumoral with intratumoral features. This approach not only surpasses models based on clinical data but also underscores the importance of comprehensive radiomic analysis in accurately characterizing breast nodules.

Background: Accurate assessment of Rheumatoid Arthritis (RA) activity remains a challenge. Multimodal photoacoustic/ultrasound (PA/US) joint imaging emerges as a novel imaging modality capable of depicting microvascularization and oxygenation levels in inflamed joints associated with RA. However, the scarcity of large-scale studies limits the exploration of correlating joint oxygenation status with disease activity. Objective: This study aimed to explore the correlation between multimodal PA/US imaging scores and RA disease activity, assessing its clinical applicability in managing RA. Methods: In this study, we recruited 111 patients diagnosed with RA and conducted examinations of seven small joints on their clinically dominant side using a PA/US imaging system. The PA and power Doppler ultrasound (PDUS) signals were semi-quantitatively assessed using a 0–3 grading system. The cumulative scores for PA and PDUS across these seven joints (PA-sum and PDUS-sum) were calculated. Relative oxygen saturation (So2) values of inflamed joints on the clinically dominant side were measured, and categorized into four distinct PA+So2 patterns. The correlation between PA/US imaging scores and disease activity indices was systematically evaluated. Results: Analysis of 777 small joints in 111 patients revealed that the PA-sum scores exhibited a strong positive correlation with standard clinical scores for RA, including DAS28 [ESR] (ρ = 0.682), DAS28 [CRP] (ρ = 0.683), CDAI (ρ = 0.738), and SDAI (ρ = 0.739), all with p < 0.001. These correlations were superior to those of the PDUS-sum scores (DAS28 [ESR] ρ = 0.559, DAS28 [CRP] ρ = 0.555, CDAI ρ = 0.575, SDAI ρ = 0.581, p < 0.001). Significantly, in patients with higher PA-sum scores, notable differences were observed in the erythrocyte sedimentation rate (ESR) (p < 0.01) and swollen joint count 28 (SJC28) (p < 0.01) between hypoxia and intermediate groups. Notably, RA patients in the hypoxia group exhibited higher clinical scores in certain clinical indices. Conclusion: Multi-modal PA/US imaging introduces potential advancements in RA assessment, especially regarding So2 evaluations in synovial tissues and associated PA scores. However, further studies are warranted, particularly with more substantial sample sizes and in multi-center settings. Summary: This study utilized multi-modal PA/US imaging to analyze Rheumatoid Arthritis (RA) patients' synovial tissues and affected joints. When juxtaposed with traditional PDUS imaging, the PA approach demonstrated enhanced sensitivity, especially concerning detecting small vessels in thickened synovium and inflamed tendon sheaths. Furthermore, correlations between the derived PA scores, PA+So2 patterns, and standard clinical RA scores were observed. These findings suggest that multi-modal PA/US imaging could be a valuable tool in the comprehensive assessment of RA, offering insights not only into disease activity but also into the oxygenation status of synovial tissues. However, as promising as these results are, further investigations, especially in larger and diverse patient populations, are imperative. Key points: ⸸ Multi-modal PA/US Imaging in RA: This novel technique was used to assess the So2 values in synovial tissues and determine PA scores of affected RA joints.⸸ Correlation significantly with Clinical RA Scores: Correlations significantly were noted between PA scores, PA+So2 patterns, and standard clinical RA metrics, hinting at the potential clinical applicability of the technique.

IntroductionNatural plants are valuable resources for exploring new bioactive compounds. Artemisia vulgaris L. is a traditional Chinese medicinal herb that has been historically used for treating multiple diseases. Active compounds isolated and extracted from A. vulgaris L. typically possess immunomodulatory and anti-inflammatory properties. Artemvulactone E (AE) is a new sesquiterpene lactone isolated and extracted from A. vulgaris L. with unclear biological activities.MethodsThe immunoregulatory effects of AE on macrophages were assessed by ELISA, RT-qPCR, immunofluorescence, and western blot assay. The effect of AE on lipopolysaccharide (LPS) -relates signaling pathways was examined by western blot assay. In zebrafish models, the larvae were yolk-microinjected with LPS to establish inflammation model and the effect of AE was evaluated by determining the survival rate, heart rate, yolk sac edema size, neutrophils and macrophages infiltration of zebrafish. The interaction between AE and Toll-like receptor 4 (TLR4) was examined by molecular docking and dynamic stimulation.ResultsAE reduced the expression and secretion of pro-inflammatory cytokines (TNF-α and IL-6), inflammatory mediators iNOS and COX-2, as well as decreases the production of intracellular NO and ROS in LPS-stimulated macrophages. In addition, AE exerted its anti-inflammatory effect synergistically by inhibiting MAPK/JAK/STAT3-NF-κB signaling pathways. Furthermore, AE enhanced the survival rate and attenuated inflammatory response in zebrafish embryos treated with LPS. Finally, the molecular dynamics results indicate that AE forms stable complexes with LPS receptor TLR4 through the Ser127 residue, thus completely impairing the subsequent activation of MAPK-NF-κB signaling.ConclusionAE exhibits notable anti-inflammatory activity and represents as a potential agent for treating inflammation-associated diseases.

Abstract Purpose To assess the diagnostic performance of Ultrasound Attenuation Analysis (USAT) in the diagnosis and grading of hepatic steatosis in patients with non-alcoholic fatty liver disease (NAFLD) using Controlled Attenuation Parameters (CAP) as a reference. Materials and methods From February 13, 2023, to September 26, 2023, participants underwent CAP and USAT examinations on the same day. We used manufacturer-recommended CAP thresholds to categorize the stages of hepatic steatosis: stage 1 (mild) − 240 dB/m, stage 2 (moderate) − 265 dB/m, stage 3 (severe) − 295 dB/m. Receiver Operating Characteristic curves were employed to evaluate the diagnostic accuracy of USAT and determine the thresholds for different levels of hepatic steatosis. Results Using CAP as the reference, we observed that the average USAT value increased with the severity of hepatic steatosis, and the differences in USAT values among the different hepatic steatosis groups were statistically significant (p

Abstract Background The application of artificial intelligence (AI) in the ultrasound (US) diagnosis of breast cancer (BCa) is increasingly prevalent. However, the impact of US-probe frequencies on the diagnostic efficacy of AI models has not been clearly established. Objectives To explore the impact of using US-video of variable frequencies on the diagnostic efficacy of AI in breast US screening. Methods This study utilized different frequency US-probes (L14: frequency range: 3.0-14.0 MHz, central frequency 9 MHz, L9: frequency range: 2.5-9.0 MHz, central frequency 6.5 MHz and L13: frequency range: 3.6-13.5 MHz, central frequency 8 MHz, L7: frequency range: 3-7 MHz, central frequency 4.0 MHz, linear arrays) to collect breast-video and applied an entropy-based deep learning approach for evaluation. We analyzed the average two-dimensional image entropy (2-DIE) of these videos and the performance of AI models in processing videos from these different frequencies to assess how probe frequency affects AI diagnostic performance. Results The study found that in testing set 1, L9 was higher than L14 in average 2-DIE; in testing set 2, L13 was higher in average 2-DIE than L7. The diagnostic efficacy of US-data, utilized in AI model analysis, varied across different frequencies (AUC: L9 > L14: 0.849 vs. 0.784; L13 > L7: 0.920 vs. 0.887). Conclusion This study indicate that US-data acquired using probes with varying frequencies exhibit diverse average 2-DIE values, and datasets characterized by higher average 2-DIE demonstrate enhanced diagnostic outcomes in AI-driven BCa diagnosis. Unlike other studies, our research emphasizes the importance of US-probe frequency selection on AI model diagnostic performance, rather than focusing solely on the AI algorithms themselves. These insights offer a new perspective for early BCa screening and diagnosis and are of significant for future choices of US equipment and optimization of AI algorithms.

Abstract Eosinophils are a group of granulocytes well known for their capacity to protect the host from parasites and regulate immune function. Diverse biological roles for eosinophils have been increasingly identified, but the developmental pattern and regulation of the eosinophil lineage remain largely unknown. Herein, we utilize the zebrafish model to analyze eosinophilic cell differentiation, distribution, and regulation. By identifying eslec as an eosinophil lineage-specific marker, we establish a Tg(eslec:eGFP) reporter line, which specifically labeled cells of the eosinophil lineage from early life through adulthood. Spatial-temporal analysis of eslec + cells demonstrates their organ distribution from larval stage to adulthood. By single-cell RNA-Seq analysis, we decipher the eosinophil lineage cells from lineage-committed progenitors to mature eosinophils. Through further genetic analysis, we demonstrate the role of Cebp1 in balancing neutrophil and eosinophil lineages, and a Cebp1-Cebpβ transcriptional axis that regulates the commitment and differentiation of the eosinophil lineage. Cross-species functional comparisons reveals that zebrafish Cebp1 is the functional orthologue of human C/EBPεP27 in suppressing eosinophilopoiesis. Our study characterizes eosinophil development in multiple dimensions including spatial-temporal patterns, expression profiles, and genetic regulators, providing for a better understanding of eosinophilopoiesis.

Objectives Triglyceride (TG), triglyceride-glucose index (TyG), body mass index (BMI), TyG-BMI and triglyceride to high-density lipoprotein ratio (TG/HDL) have been reported to be reliable predictors of non-alcoholic fatty liver disease. However, there are few studies on potential predictors of non-alcoholic fatty pancreas disease (NAFPD). Our aim was to evaluate these and other parameters for predicting NAFPD.Design Cross-sectional study design.Setting Physical examination centre of a tertiary hospital in China.Participants This study involved 1774 subjects who underwent physical examinations from January 2016 to September 2016.Primary and secondary outcome measures From each subject, data were collected for 13 basic physical examination and blood biochemical parameters: age, weight, height, BMI, TyG, TyG-BMI, high-density lipoprotein (HDL), low-density lipoprotein, total cholesterol, TG, fasting plasma glucose, TG/HDL and uric acid. NAFPD was diagnosed by abdominal ultrasonography. A logistic regression model with a restricted cubic spline was used to evaluate the relationship between each parameter and NAFPD. The receiver operating characteristic (ROC) curve was used to calculate the area under the curve for each parameter.Results HDL was negatively correlated with NAFPD, height was almost uncorrelated with NAFPD and the remaining 11 parameters were positively correlated with NAFPD. ROC curve showed that weight-related parameters (weight, BMI and TyG-BMI) and TG-related parameters (TyG, TG and TG/HDL) had high predictive values for the identification of NAFPD. The combinations of multiple parameters had a better prediction effect than a single parameter. All the predictive effects did not differ by sex.Conclusions Weight-related and TG-related parameters are good predictors of NAFPD in all populations. BMI showed the greatest predictive potential. Multiparameter combinations appear to be a good way to predict NAFPD.

Abstract Background This retrospective study aims to validate the effectiveness of artificial intelligence (AI) to detect and classify non-mass breast lesions (NMLs) on ultrasound (US) images. Methods A total of 228 patients with NMLs and 596 volunteers without breast lesions on US images were enrolled in the study from January 2020 to December 2022. The pathological results served as the gold standard for NMLs. Two AI models were developed to accurately detect and classify NMLs on US images, including DenseNet121_448 and MobileNet_448. To evaluate and compare the diagnostic performance of AI models, the area under the curve (AUC), accuracy, specificity and sensitivity was employed. Results A total of 228 NMLs patients confirmed by postoperative pathology with 870 US images and 596 volunteers with 1003 US images were enrolled. In the detection experiment, the MobileNet_448 achieved the good performance in the testing set, with the AUC, accuracy, sensitivity, and specificity were 0.999 (95%CI: 0.997-1.000),96.5%,96.9% and 96.1%, respectively. It was no statistically significant compared to DenseNet121_448. In the classification experiment, the MobileNet_448 model achieved the highest diagnostic performance in the testing set, with the AUC, accuracy, sensitivity, and specificity were 0.837 (95%CI: 0.990-1.000), 70.5%, 80.3% and 74.6%, respectively. Conclusions This study suggests that the AI models, particularly MobileNet_448, can effectively detect and classify NMLs in US images. This technique has the potential to improve early diagnostic accuracy for NMLs.

Abstract Nonsteroidal anti-inflammatory drugs compose one of the most widely used classes of medications, but the risks for early development remain controversial, especially in the nervous system. Here, we utilized zebrafish larvae to assess the potentially toxic effects of nonsteroidal anti-inflammatory drugs and found that sulindac can selectively induce apoptosis of GABAergic neurons in the brains of zebrafish larvae brains. Zebrafish larvae exhibit hyperactive behaviour after sulindac exposure. We also found that akt1 is selectively expressed in GABAergic neurons and that SC97 (an Akt1 activator) and exogenous akt1 mRNA can reverse the apoptosis caused by sulindac. Further studies showed that sulindac binds to retinoid X receptor alpha (RXRα) and induces autophagy in GABAergic neurons, leading to activation of the mitochondrial apoptotic pathway. Finally, we verified that sulindac can lead to hyperactivity and selectively induce GABAergic neuron apoptosis in mice. These findings suggest that excessive use of sulindac may lead to early neurodevelopmental toxicity and increase the risk of hyperactivity, which could be associated with damage to GABAergic neurons.

t(1;19)(q23;p13) is one of the most common translocation genes in childhood acute lymphoblastic leukemia (ALL) and is also present in acute myeloid leukemia (AML) and mixed-phenotype acute leukemia (MPAL). This translocation results in the formation of the oncogenic E2A-PBX1 fusion protein, which contains a trans-activating domain from E2A and a DNA-binding homologous domain from PBX1. Despite its clear oncogenic potential, the pathogenesis of E2A-PBX1 fusion protein is not fully understood (especially in leukemias other than ALL), and effective targeted clinical therapies have not been developed. To address this, we established a stable and heritable zebrafish line expressing human E2A-PBX1 (hE2APBX1) for high-throughput drug screening. Blood phenotype analysis showed that hE2APBX1 expression induced myeloid hyperplasia by increasing myeloid differentiation propensity of hematopoietic stem cells (HSPCs) and myeloid proliferation in larvae, and progressed to AML in adults. Mechanistic studies revealed that hE2A-PBX1 activated the TNF/IL-17/MAPK signaling pathway in blood cells and induced myeloid hyperplasia by upregulating the expression of the runx1. Interestingly, through high-throughput drug screening, three small molecules targeting the TNF/IL-17/MAPK signaling pathway were identified, including OUL35, KJ-Pyr-9, and CID44216842, which not only alleviated the hE2A-PBX1- induced myeloid hyperplasia in zebrafish but also inhibited the growth and oncogenicity of human pre-B ALL cells with E2A-PBX1. Overall, this study provides a novel hE2A-PBX1 transgenic zebrafish leukemia model and identifies potential targeted therapeutic drugs, which may offer new insights into the treatment of E2A-PBX1 leukemia.

Interleukin-34 (IL-34), a relatively newly identified cytokine, binds to colony stimulating factor 1 receptor (CSF-1R) to activate several signaling pathways. It thus performs essential roles in immune surveillance, the inflammatory response, cell proliferation, and cell migration. In this study, the homologues of IL-34 (designated ‘CaIL-34′) and two types of CSF-1R (designated ‘CaCSF-1R1′ and ‘CaCSF-1R2′) are identified and described in the snakehead, Channa argus. Analysis of the amino acid sequence revealed that CaIL-34 has relatively conservative structural characteristics. CaCSF-1R1 and CaCSF-1R2 both contain four immunoglobulin-like domains and a tyrosine kinase domain. Their transcriptions were detected in all the tissues examined in healthy individuals, with relatively high transcription levels in immune-related tissues. The two major bacterial pathogens, Nocardia seriolae and Aeromonas schubertii, both induced their transcription in the head kidney and spleen tissues of snakeheads in vivo. The three classical pathogen-associated molecular patterns, lipopolysaccharide (LPS), lipoteichoic acid (LTA), and polyinosinic–polycytidylic acid (poly[I:C]), and six recombinant cytokines of the snakehead, recombinant tumor necrosis factor-α1 (TNF-α1), TNF-α2, IL-1β, IL-18, IL-20, and IL-21, all increased their transcription in the head kidney leukocytes (HKLs) in vitro, albeit at different levels. Recombinant CaIL-34 (rCaIL-34) protein was expressed in a prokaryotic system. After stimulation with rCaIL-34, the transcription of the IL-1β, TNF-α1, TNF-α2, and interferon-γ (IFN-γ) genes, and endogenous CaIL-34, CaCSF-1R1, and CaCSF-1R2, was enhanced in HKLs, and HKL proliferation was promoted. A subcellular localization analysis showed that CaIL-34 was exclusively located in the cytoplasm of HEK293T cells. The overexpression of CaIL-34 activated the nuclear translocation of nuclear factor kappa B (NF-κB) and activator protein 1 (AP-1) signaling, albeit to different levels. In summary, CaIL-34 and two types of CSF-1R of C. argus participate strongly in its antibacterial responses, leukocyte proliferation, and the activation of NF-κB and AP-1 signaling.

Purpose: The objective of this research was to investigate the efficacy of various parameter combinations of Convolutional Neural Networks (CNNs) models, namely MobileNet and DenseNet121, and different input image resolutions (REZs) ranging from 64×64 to 512×512 pixels, for diagnosing breast cancer. Materials and methods: During the period of June 2015 to November 2020, two hospitals were involved in the collection of two-dimensional ultrasound breast images for this retrospective multicenter study. The diagnostic performance of the computer models MobileNet and DenseNet 121 was compared at different resolutions. Results: The results showed that MobileNet had the best breast cancer diagnosis performance at 320×320pixel REZ and DenseNet121 had the best breast cancer diagnosis performance at 448×448pixel REZ. Conclusion: Our study reveals a significant correlation between image resolution and breast cancer diagnosis accuracy. Through the comparison of MobileNet and DenseNet121, it is highlighted that lightweight neural networks (LW-CNNs) can achieve model performance similar to or even slightly better than large neural networks models (HW-CNNs) in ultrasound images, and LW-CNNs' prediction time per image is lower.

Abstract Myelodysplastic syndromes (MDS) are characterized by daunting genetic heterogeneity and a high risk of leukemic transformation, which presents great challenges for clinical treatment. To identify new chemicals for MDS, we screened a panel of FDA-approved drugs and verified the neutrophil hyperplasia inhibiting role of 17β-estradiol (E2, a natural estrogen) in several zebrafish MDS models (pu.1 G242D/G242D , irf8 Δ57Δ/57 and c-myb hyper ). However, the protective mechanism of estrogen in the development of hematological malignancies remains to be explored. Here, analyzing the role of E2 in the development of each hematopoietic lineage, we found that E2 exhibited a specific neutrophil inhibiting function. This neutrophil inhibitory function of E2 is attributed to its down-regulation of c-myb, which leads to accelerated apoptosis and decreased proliferation of neutrophils. We further showed that knockdown of hif1α could mimic the neutrophil inhibiting role of E2, and hif1α overexpression could reverse the protective function of E2. Collectively, our findings highlight the protective role of E2 on MDS by inhibiting hif1α-c-myb pathway, suggesting that E2 is a promising and effective drug for hematopoietic tumors associated with abnormal neutrophil hyperplasia.

Abstract Background Alcoholism is a well-known risk factor for liver injury and is one of the major causes of hepatic steatosis worldwide. Although many drugs have been reported to have protective effects against acute alcohol-induced hepatotoxicity, there is limited available treatment for alcoholic liver disease (ALD), indicating an urgent need for effective therapeutic options. Herein, we first reported the protective effects of magnesium isoglycyrrhizinate (MgIG) on acute alcohol-induced hepatic steatosis and its related mechanisms in a zebrafish model. Methods Alcohol was administered directly to embryo medium at 5 days post-fertilization (dpf) for up to 32 h. MgIG was given to the larvae 2 h before the administration of alcohol and then cotreated with alcohol starting at 5 dpf. Oil red O staining was used to determine the incidence of steatosis, and pathological features of the liver were assessed by hematoxylin–eosin staining. Biological indexes, total cholesterol (TC) and triacylglycerol (TG) were detected in the livers of zebrafish larvae. Morphological changes in the livers of zebrafish larvae were observed using liver-specific EGFP transgenic zebrafish (Tg(lfabp10a:eGFP)). The expression levels of critical molecules related to endoplasmic reticulum (ER) stress and lipid metabolism were detected by qRT–PCR, whole-mount in situ hybridization and western blotting. Results Alcohol-treated larvae developed hepatomegaly and steatosis after 32 h of exposure. We found that MgIG improved hepatomegaly and reduced the incidence of steatosis in a dose-dependent manner by oil red O staining and diminished deposits of alcohol-induced fat droplets by histologic analysis. Moreover, MgIG significantly decreased the levels of TC and TG in the livers of zebrafish larvae. Furthermore, the expression levels of critical genes involved in ER stress (atf6, irela, bip, chop) and the key enzymes regulating lipid metabolism (acc1, fasn, hmgcs1 and hmgcra) were significantly higher in the alcohol-treated group than in the control group. However, in the MgIG plus alcohol-treated group, the expression of these genes was markedly decreased compared with that in the alcohol-treated group. Whole-mount in situ hybridization and western blotting also showed that MgIG had an effect on the expression levels of critical genes and proteins involved in lipid metabolism and ER stress. Our results revealed that MgIG could markedly regulate these genes and protect the liver from ER stress and lipid metabolism disorders. Conclusions Our study is the first to demonstrate that MgIG could protect the liver from acute alcohol stimulation by ameliorating the disorder of lipid metabolism and regulating ER stress in zebrafish larvae.

Nocardia seriolae, an intracellular gram-positive pathogen, is prone to infecting immunocompromised and surface-damaged fish, causing serious losses to the aquaculture industry. Although a previous study has demonstrated that N. seriolae infects macrophages, the persistence of this bacterium in macrophages has not been well characterized. To address this gap, we used the macrophage cell line RAW264.7, to investigate the interactions between N. seriolae and macrophages and deciphered the intracellular survival mechanism of N. seriolae. Confocal and light microscopy revealed that N. seriolae entered macrophages 2 hours post-inoculation (hpi), were phagocytosed by macrophages at 4–8 hpi, and induced the formation of multinucleated macrophages by severe fusion at 12 hpi. Flow cytometry, evaluation of mitochondrial membrane potential, release of lactate dehydrogenase, and observation of the ultrastructure of macrophages revealed that apoptosis was induced in the early infection stage and inhibited in the middle and later periods of infection. Additionally, the expression of Bcl-2, Bax, Cyto-C, Caspase-3, Capase-8, and Caspase-9 was induced at 4 hpi, and then decreased at 6–8 hpi, illustrating that N. seriolae infection induces the activation of extrinsic and intrinsic apoptotic pathways in macrophages, followed by the inhibition of apoptosis to survive inside the cells. Furthermore, N. seriolae inhibits the production of reactive oxygen species and releases large amounts of nitric oxide, which persists in macrophages during infection. The present study provides the first comprehensive insight into the intracellular behavior of N. seriolae and its apoptotic effect on macrophages and may be important for understanding the pathogenicity of fish nocardiosis.

The increasing abundance of antibiotic resistance genes (ARGs), which are regarded as new pollutants, has raised public health concern. The use of antibiotics in aquaculture has promoted the evolution and spread of ARGs. The occurrence and abundance of ARGs in aquaculture has attracted extensive attention. However, the distribution and transmission of ARGs in aquaculture require further study. This study analyzed water and sediment from intensive culture of hybrid snakehead fish farm in Zhongshan, South China. Twenty-two types of ARGs were detected in all environmental samples. The relative abundance of sulfonamide resistance genes (sul1 and sul2) was the highest, ranging from 3.37×10-2 to 8.53×100 copies/16S rRNA gene. High occurrence of quinolones, phenicols, tetracycline resistance genes, and class 1 integrase gene (intI1) was also observed in pond water samples. This implies that pond water is one of the main reservoirs and origins of ARGs in the aquatic environment. Proteobacteria was the most abundant phylum in all the environmental samples, and its relative abundance ranged from 24.05% to 41.84%. Network and canonical correspondence analyses showed that a high abundance of ARGs (int1, sul1, sul2, qacEΔ1, aac6, and oqxA) was positively correlated with Proteobacteria, Cyanobacteria, and Bacteroidetes, and the abundance of Proteobacteria, Actinobacteria, and Patescibacteria was positively correlated with environmental factors (sulfide, nitrite nitrogen, pH, free chlorine, and ammonia nitrogen). These findings demonstrate the prevalence and persistence of ARGs in intensive fish farming in southern China. This suggests that ARG levels and microbiological community composition in aquaculture should be conventionally determined to assess potential risks to public health.

Abstract Hematopoietic stem cells (HSCs) possess the potential for self-renew and the capacity, throughout life, to differentiate into all blood cell lineages. Yet, the mechanistic basis for HSC development remains largely unknown. In this study, we characterized a zebrafish smu471 mutant with hematopoietic stem/progenitor cell (HSPC) defects and found that sart3 was the causative gene. RNA expression profiling of the sart3 smu471 mutant revealed spliceosome and p53 signaling pathway to be the most significantly enriched pathways in the sart3 smu471 mutant. Knock down of p53 rescued HSPC development in the sart3 smu471 mutant. Interestingly, the p53 inhibitor, mdm4, had undergone an alternative splicing event in the mutant. Restoration of mdm4 partially rescued HSPC deficiency. Thus, our data suggest that HSPC proliferation and maintenance require sart3 to ensure the correct splicing and expression of mdm4, so that the p53 pathway is properly inhibited to prevent definitive hematopoiesis failure. This study expands our knowledge of the regulatory mechanisms that impact HSPC development and sheds light on the mechanistic basis and potential therapeutic use of sart3 in spliceosome-mdm4-p53 related disorders.

BackgroundDisorder of consciousness (DOC) is frequent in patients with stroke, which is the second most common cause of death and a leading cause of disability. Acupuncture has been used as a curative method for DOC treatment in China. Nevertheless, no critical systematic review of acupuncture's effect on DOC has been published. This review aims to evaluate the present evidence regarding the efficacy of acupuncture for DOC after stroke.MethodsSeven databases were searched from their inception to November 1, 2021, containing three English databases (PubMed, Embase, and Cochrane Central Register of Controlled Trials) and four Chinese databases (CNKI, CBM, VIP, and Wanfang Database). The primary outcomes comprise the Glasgow Coma Scale (GCS) and Glasgow Outcome Scale (GOS) before and after treatment. Secondary outcomes involve resuscitation rate, resuscitation time, and adverse events. Data synthesis was calculated by RevMan (V.5.4.1) software. According to the Cochrane Handbook, methodological quality was assessed with the risk of bias tool 2.0 (RoB2).ResultsSeventeen studies containing 1,208 patients were eventually included in our review. Overall, most trials were rated as high or had some concerns regarding the risk of bias. GCS was reported in 16 trials, and a meta-analysis showed that GCS improvement in the acupuncture group was greater than in the non-acupuncture group (MD 1.45, 95% CI 0.94–1.97, P < 0.0001). One trial reported that GOS improvement in the acupuncture plus medication group was greater than in the medication group (MD 0.58, 95% CI 0.11–1.05, P = 0.01). Another study reported that acupuncture plus medication was statistically more effective in shortening resuscitation time than medication alone (MD−0.89, 95% CI −1.53 to −0.25, P = 0.006). Four trials reported that the resuscitation rate in the acupuncture group was higher than without acupuncture intervention (RR 1.68, 95% CI 1.30–2.18, I2 0%, P = 0.39). Adverse events were reported in two studies, with one case in the acupuncture group suffering from subcutaneous hematoma.ConclusionAcupuncture may improve consciousness level, increase the resuscitation rate, and shorten resuscitation time for post-stroke patients with DOC. Adverse events from acupuncture were rare, tolerable, and recoverable. However, the results should be interpreted cautiously, and more rigorous RCTs with better methodology are warranted.Systematic review registrationhttps://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=289802, identifier 289802.

Although numerous epidemiological studies have demonstrated a relationship between environmental factors and chronic diseases, there is a lack of comprehensive population health vulnerability assessment studies from the perspective of environmental exposure, population sensitivity and adaptation on a regional scale. To address this gap, this study focused on six high-mortality chronic diseases in China and constructed an exposure–sensitivity–adaptability framework-based index system using multivariate data. The constructed system effectively estimated health vulnerability for the chronic diseases. The R-square between vulnerability and mortality rates for respiratory diseases and malignant tumors exceeded 0.7 and was around 0.6 for the other four chronic diseases. In 2020, Chongqing exhibited the highest vulnerability to respiratory diseases. For heart diseases, vulnerability values exceeding 0.5 were observed mainly in northern and northeastern provinces. Vulnerability values above 0.5 were observed in Jiangsu, Shanghai, Tianjin, Shandong and Liaoning for cerebrovascular diseases and malignant tumors. Shanghai had the highest vulnerability to endogenous metabolic diseases, and Tibet exhibited the highest vulnerability to digestive system diseases. The main related factor analysis results show that high temperature and humidity, severe temperature fluctuations, serious air pollution, high proportion of middle-aged and elderly population, as well as high consumption of aquatic products, red meat and eggs increased health vulnerability, while increasing per capita educational resources helped reduce vulnerability.

With the increasing growth of cyber-attack incidences, it is important to develop innovative and effective techniques to assess and defend networked systems against cyber attacks. One of the well-known techniques for this is performing penetration testing which is carried by a group of security professionals (i.e, red team). Penetration testing is also known to be effective to find existing and new vulnerabilities, however, the quality of security assessment can be depending on the quality of the red team members and their time and devotion to the penetration testing. In this paper, we propose a novel automation framework for cyber-attacks generation named `HARMer' to address the challenges with respect to manual attack execution by the red team. Our novel proposed framework, design, and implementation is based on a scalable graphical security model called Hierarchical Attack Representation Model (HARM). (1) We propose the requirements and the key phases for the automation framework. (2) We propose security metrics-based attack planning strategies along with their algorithms. (3) We conduct experiments in a real enterprise network and Amazon Web Services. The results show how the different phases of the framework interact to model the attackers' operations. This framework will allow security administrators to automatically assess the impact of various threats and attacks in an automated manner.

The science-practice gap has recently been discussed as a critical challenge restricting sustainable growth and development in all facets of our society, including explorations of Recreation Ecosystem Services (RES). To better explore how well the scientific study of RES and its application are connected, this paper aims to synthesize empirical evidence based on an in-depth and systematic literature review. We found that studies of RES have not effectively transformed into the decision-making and long-term planning of our cities. From 2005 to 2020, only 13% of studies referred to specific applications, and about 40% of papers mentioned no applications or practical implications for their research. However, RES research has many potential applications, which can be categorised into six main aspects. In terms of non-spatial improvement: Improved monetary benefits (40%), non-monetary benefits (30%); in terms of spatial improvement: space with high recreational potential or degradation (7%), the relation between supply and demand (7%); and Cross-service governance (16%). After combining the results of various studies, we developed a framework starting from applicable problems and their solutions, which can incorporate the outcomes of RES research while systematically narrowing down the research questions and methods. The framework offers a starting point for further research that can modify and improve in bridging science-practice gaps in RES studies.

Abstract Background Chronic stress is well known to promote tumor progression, however, little is known whether chronic stress-mediated regulation of osteoblasts contributes to the migration and invasion of metastatic cancer cells. Methods The proliferation, migration and invasion of prostate cancer cells were assessed by CCK-8 and transwell assay. HIF-1α expression of osteoblasts and epithelial-mesenchymal transition (EMT) markers of prostate cancer cells were examined by Western blot. The mRNA level of cytokines associated with bone metastasis in osteoblasts and EMT markers in PC-3 and DU145 cells were performed by qRT-PCR. Functional rescue experiment of cells were performed by using siRNA, plasmid transfection and inhibitor treatment. Results Isoproterenol (ISO), a pharmacological surrogate of sympathetic nerve activation induced by chronic stress, exhibited no direct effect on migration and invasion of PC-3 and DU145 prostate cancer cells. Whereas, osteoblasts pretreated with ISO promoted EMT, migration and invasion of PC-3 and DU145 cells, which could be inhibited by β2AR inhibitor. Mechanistically, ISO increased the secretion of CXCL12 via the β2AR-HIF-1α signaling in osteoblasts. Moreover, overexpression of HIF-1α osteoblasts promoted migration and invasion of PC-3 and DU145 cells, which was inhibited by addition of recombinant knockdown of CXCR4 in PC-3 and DU145 cells, and inhibiting CXCL12-CXCR4 signaling with LY2510924 blunted the effects of osteoblasts in response to ISO on EMT and migration as well as invasion of PC-3 and DU145 cells. Conclusions These findings demonstrated that β2AR-HIF-1α-CXCL12 signaling in osteoblasts facilitates migration and invasion as well as EMT of prostate cancer cells, and may play a potential role in affecting bone metastasis of prostate cancer.

Abstract Background Experimental and clinical trials have demonstrated the efficiency of bone marrow-derived mesenchymal stromal/stem cells (bMSCs) in the treatment of myocardial infarction. However, after intravenous injection, the ineffective migration of engrafted bMSCs to the hearts remains an obstacle, which has an undesirable impact on the efficiency of cell-based therapy. Therefore, we attempted to identify a marker that could distinguish a subpopulation of bMSCs with a promising migratory capacity. Methods Here, CD51-negative and CD51-positive cells were isolated by flow cytometry from Ter119−CD45−CD31−bMSCs and cultured in specifically modified medium. The proliferation ability of the cells was evaluated by 5-ethynyl-2′-deoxyuridine (EdU) staining or continuously monitored during culture, and the differentiation potential was assessed by culturing the cells in the appropriate conditioned media. Wound healing assays, transwell assays and quantitative polymerase chain reaction (qPCR) were used to measure the migratory ability. The mice were subjected to a sham operation or myocardial infarction (MI) by permanently occluding the coronary artery, and green fluorescent protein (GFP)-labelled cells were transplanted into the mice via intravenous infusion immediately after MI. Heart function was measured by echocardiography; infarct myocardium tissues were detected by triphenyl tetrazolium chloride (TTC) staining. Additionally, immunofluorescence staining was used to verify the characteristics of CD51+bMSCs and inflammatory responses in vivo. Statistical comparisons were performed using a two-tailed Student’s t test. Results In this study, the isolated CD51−bMSCs and CD51+bMSCs, especially the CD51+ cells, presented a favourable proliferative capacity and could differentiate into adipocytes, osteocytes and chondrocytes in vitro. After the cells were transplanted into the MI mice by intravenous injection, the therapeutic efficiency of CD51+bMSCs in improving left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) was better than that of CD51−bMSCs. Compared with CD51−bMSCs, CD51+bMSCs preferentially migrated to and were retained in the infarcted hearts at 48 h and 8 days after intravenous injection. Accordingly, the migratory capacity of CD51+bMSCs exceeded that of CD51−bMSCs in vitro, and the former cells expressed higher levels of chemokine receptors or ligands. Interestingly, the retained CD51+bMSCs retained in the myocardium possessed proliferative potential but only differentiated into endothelial cells, smooth muscle cells, fibroblasts or cardiomyocytes. Transplantation of CD51+bMSCs partially attenuated the inflammatory response in the hearts after MI, while the potential for inflammatory suppression was low in CD51−bMSC-treated mice. Conclusions These findings indicated that the CD51-distinguished subpopulation of bMSCs facilitated proliferation and migration both in vitro and in vivo, which provided a novel cell-based strategy to treat acute MI in mice by intravenous injection.

Summary: Type IV mucolipidosis (ML-IV) is a neurodegenerative lysosome storage disorder caused by mutations in the MCOLN1 gene. However, the cellular and molecular bases underlying the neuronal phenotypes of ML-IV disease remain elusive. Using a forward genetic screening, we identified a zebrafish mutant, biluo, that harbors a hypomorphic mutation in mcoln1a, one of the two zebrafish homologs of mammalian MCOLN1. The mcoln1a-deficient mutants display phenotypes partially recapitulating the key features of ML-IV disorder, including the accumulation of enlarged late endosomes in microglia and aberrant neuronal activities in both spontaneous and visual-evoking conditions in optic tectal neurons. We further show that the accumulation of enlarged late endosomes in microglia is caused by the impairment of late endosome and lysosome fusion and the aberrant neuronal activities can be partially rescued by the reconstitution of Mcoln1a function in microglia. Our findings suggest that dysregulation of microglial function may contribute to the development and progression of ML-IV disease. : Cellular Neuroscience; Clinical Neuroscience; Model Organism; Neuroscience Subject Areas: Cellular Neuroscience, Clinical Neuroscience, Model Organism, Neuroscience

BackgroundPostoperative delirium (POD) is a common complication after orthopedic surgery in elderly patients. The elderly may experience drastic changes in autonomic nervous system (ANS) activity and circadian rhythm disorders after surgery. Therefore, we intend to explore the relationship between postoperative long-term heart rate (HR) variability (HRV), as a measure of ANS activity and circadian rhythm, and occurrence of POD in elderly patients.MethodsThe study population of this cohort was elderly patients over 60 years of age who scheduled for orthopedic surgery under spinal anesthesia. Patients were screened for inclusion and exclusion criteria before surgery. Then, participants were invited to wear a Holter monitor on the first postoperative day to collect 24-h electrocardiographic (ECG) data. Parameters in the time domain [the standard deviation of the normal-to-normal (NN) intervals (SDNN), mean of the standard deviations of all the NN intervals for each 5-min segment of a 24-h HRV recording (SDNNI), and the root mean square of successive differences of the NN intervals (RMSSD)] and frequency domain [heart rate (HR), high frequency (HF), low frequency (LF), very low frequency (VLF), ultra low frequency (ULF), and total power (TP)] were calculated. Assessment of delirium was performed daily up to the seventh postoperative day using the Chinese version of the 3-Min Diagnostic Interview for CAM-defined Delirium (3D-CAM). The relationship between HRV and POD, as well as the association between HRV and duration of POD, was assessed.ResultsOf the 294 cases that finally completed the follow-up, 60 cases developed POD. Among the HRV parameters, SDNNI, VLF, and ULF were related to the occurrence of POD. After adjustment for potential confounders, the correlation between HRV indices and POD disappeared. Through stratified analysis, two significant negative correlations emerged: ULF in young-old participants and SDNNI, VLF, and ULF in male patients.ConclusionThe lower HRV parameters may be related to the occurrence of POD, and this correlation is more significant in young-old and male patients. ANS disorders and rhythm abnormalities reflected by HRV changes may represent a possible mechanism that promotes POD.

Immersion vaccination relies on the response of fish mucosa-associated lymphoid tissues, the Crucian carp (Carassius auratus) and Grouper (Epinephelus coioides) were researched in this paper to examine local mucosal immune responses and associated humoral system responses following immersion vaccination. We administered 1.5 × 107 CFU/ml formalin-inactivated Vibrio harveyi cells and measured mucus and serum antibody titers as well as IgM, MHC II mRNA levels in immune organs. The mucosal antibody response preceded the serum response indicating a role for local mucosal immunity in immersion vaccination. IgM and MHC II mRNA levels were relatively greater for the spleen and head kidney indicating the importance and central position of systemic immunity. Expression levels were also high for the gills while skin levels were the lowest. IgM and MHC II mRNA levels were altered over time following vaccination and the hindgut, liver and spleen were similar indicating a close relationship, so the absolute value of r is used to analyze the correlation among different organs immunized. It can be inferred the existence of an internal immune molecular mechanism for Immune synergy hindgut-liver-spleen, from the peak time (14th day), the relative ratio of genes expression in the same tissues between the immunized grouper and the control group (26 times), and Pearson correlation coefficient (0.8

The large area estimation of forest canopy closure (FCC) using remotely sensed data is of high interest in monitoring forest changes and forest health, as well as in assessing forest ecological services. The accurate estimation of FCC over the regional or global scale is challenging due to the difficulty of sample acquisition and the slow processing efficiency of large amounts of remote sensing data. To address this issue, we developed a novel bounding envelope methodology based on vegetation indices (BEVIs) for determining vegetation and bare soil endmembers using the normalized differences vegetation index (NDVI), modified bare soil index (MBSI), and bare soil index (BSI) derived from Landsat 8 OLI and Sentinel-2 image within the Google Earth Engine (GEE) platform, then combined the NDVI with the dimidiate pixel model (DPM), one of the most commonly used spectral-based unmixing methods, to map the FCC distribution over an area of more than 90,000 km2. The key processing was the determination of the threshold parameter in BEVIs that characterizes the spectral boundary of vegetation and soil endmembers. The results demonstrated that when the threshold equals 0.1, the extraction accuracy of vegetation and bare soil endmembers is the highest with the threshold range given as (0, 0.3), and the estimated spatial distribution of FCC using both Landsat 8 and Sentinel-2 images were consistent, that is, the area with high canopy density was mainly distributed in the western mountainous region of Chifeng city. The verification was carried out using independent field plots. The proposed approach yielded reliable results when the Landsat 8 data were used (R2 = 0.6, RMSE = 0.13, and 1-rRMSE = 80%), and the accuracy was further improved using Sentinel-2 images with higher spatial resolution (R2 = 0.81, RMSE = 0.09, and 1-rRMSE = 86%). The findings demonstrate that the proposed method is portable among sensors with similar spectral wavebands, and can assist in mapping FCC at a regional scale while using multispectral satellite imagery.

Chronic myeloid leukemia (CML) is induced by the BCR/ABL1 oncogene, which encodes a protein tyrosine kinase. We examined the effect of direct overexpression of the human p210BCR/ABL1 oncoprotein in zebrafish. Humanized p210BCR/ABL1 protein was detectable in Tg(hsp70: p210BCR/ABL1) transgenic zebrafish embryos and adult kidney marrow. Transgenic zebrafish developed CML, which could be induced via cells transplanted into recipients. The expression of human BCR/ABL1 promoted myeloid lineages in Tg(hsp70:p210BCR/ABL1) transgenic embryos. A total of 77 of 101 (76.24%) Tg(hsp70:p210BCR/ABL1) adult transgenic zebrafish (age 6 months-1 year) developed CML. CML in zebrafish showed a triphasic phenotype, similar to that in humans, involving a chronic phase predominantly characterized by neutrophils in various degrees of maturation, an accelerated phase with an increase in blasts and immature myeloid elements, and a blast phase with >90% blasts in both the peripheral blood and kidney marrow. Tyrosine kinase inhibitors, as the standard drug treatment for human CML, effectively reduced the expanded myeloid population in Tg(hsp70:p210BCR/ABL1) transgenic embryos. Moreover, we screened a library of 171 compounds and identified ten new drugs against BCR/ABL1 kinase-dependent or -independent pathways that could also reduce lcp1+ myeloid cell numbers in Tg(hsp70:p210BCR/ABL1) transgenic embryos. In summary, we generated the first humanized zebrafish CML model that recapitulates many characteristics of human CML. This novel in vivo model will help to elucidate the mechanisms of CML disease progression and allow high-throughput drug screening of possible treatments for this disease.

This work presents a palladium-catalyzed oxime ether-directed ortho-selective benzoylation using benzoylformic acid as the acyl source with a palladium catalyst and 4CzIPN as the co-catalyst under light. Various non-symmetric benzophenone derivatives were obtained in moderate to good yields. A preliminary mechanism study revealed that the reaction proceeds through a free radical pathway.

To meet the transportation requirements of heavy-haul trains, it is necessary to consider the use of prestressed reinforced concrete ballastless tracks. Therefore, the CRTSIII SBT (China Railway Track System III-type slab ballastless track) has been considered for its convenient construction and geometric adjustment. Considering the problems existing in the base plate of CRTSIII SBT in heavy-haul applications, an indoor test and simulation were conducted to explore the cracking phenomenon of the base plate concrete under fatigue load. The reliability of the simulation was verified through the experimental results. A fatigue crack development law of the base plate structure was established. Furthermore, the influence of the load position, the thickness of self-compacting concrete and axle loads on the fatigue damage of the base plate structure were investigated. For the CRTSIII SBT subjected to heavier axle loads, it is appropriate to thicken the self-compacting concrete layer and strengthen the concrete and steel reinforcement at the plate end to reduce fatigue damage and cracks. The research results can serve as a basis for the optimization of the CRTSIII SBT base plate structure in heavy-haul railway applications.

Infectious spleen and kidney necrosis virus (ISKNV) resulted in severe systemic diseases with high morbidity and mortality in Siniperca chuatsi. Vaccination is the primary method for effective prevention and control of these diseases. The development of inactivated ISKNV vaccines made some progress, but the technique of quality evaluation is scarce. Herein, a measurement of the MCP (major capsid protein) antigen concentration for the inactivated ISKNV vaccine was developed by double-antibody sandwich ELISA. Firstly, mouse monoclonal antibodies against ISKNV particles and MCP were generated. Then, a double-antibody sandwich ELISA was developed using the monoclonal antibody 1C8 1B9 as the capture antibody and Biotin-3B12 6B3 as the detection antibody. A standard curve was generated using the MCP concentration versus OD value with the linear range of concentration of 4.69~300 ng/mL. The assay sensitivity was 0.9 ng/mL. The antigen content of three batches of inactivated ISKNV vaccines was quantitatively detected using the double-antibody sandwich ELISA. The results showed that MCP antigen contents of inactivated ISKNV vaccines were positively correlated with the viral titers. The newly established double-antibody sandwich ELISA provided a useful tool for the detection of antigen quality for ISKNV inactivated vaccines.

The machine learning has been widely used in intelligent data mining. The high-computational complexity of machine learning and huge data volume present challenges to computing platforms. Graphics processor unit (GPU) provides powerful computing support for machine learning but shows different performances under different computing scales and/or different development methods. Analysing the performance of GPUs in different application scenarios helps to improve computing performance. In this study, the matrix multiplication, which is a common and time-consuming computation operation in machine learning, is performed on different data scales and different development methods to analyse the relationship between GPU computing performance with matrix scale and development methods. The experimental data shows that the performance of GPU is not much improved compared with the central processing unit in small-scale data calculation. Also, using a high-level application programming interface for GPU development is less computing-efficient than the GPU programming language computes unified device architecture C.

Forest canopy height is an indispensable forest vertical structure parameter for understanding the carbon cycle and forest ecosystem services. A variety of studies based on spaceborne Lidar, such as ICESat, ICESat-2 and airborne Lidar, were conducted to estimate forest canopy height at multiple scales. However, while a few studies have been conducted based on ICESat-2 simulated data from airborne Lidar data, few studies have analyzed ATL08 and ATL03 products derived from the ATLAS sensor onboard ICESat-2 for regional vegetation canopy height mapping. It is necessary and promising to explore how data obtained by ICESat-2 can be applied to estimate forest canopy height. This study proposes a new means to estimate forest canopy height, defined as the mean height of trees within a given forest area, using a combination of ICESat-2 ATL08 and ATL03 data and ZY-3 satellite stereo images. Five procedures were used to estimate the forest canopy height of the city of Nanning in China: (1) Processing ground photons in a 30 m × 30 m grid; (2) Extracting a digital surface model (DSM) using ZY-3 stereo images; (3) Calculating a discontinuous canopy height model (CHM) dataset; (4) Validating the DSM and ground photon height using GEDI data; (5) Estimating the regional wall-to-wall forest canopy height product based on the backpropagation artificial neural network (BP-ANN) model and Landsat 8 vegetation indices and independent accuracy assessments with field measured plots. The validation shows a root mean square error (RMSE) of 3.34 m to 3.47 m and a coefficient of determination R2 = 0.51. The new method shows promise and can be used for large-scale forest canopy height mapping at various resolutions or in combination with other data, such as SAR images. Finally, this study analyzes resolutions and how to filter effective data when ATL08 data are directly used to generate regional or global vegetation height products, which will be the focus of future research.

Accurate information regarding forest volume plays an important role in estimating afforestation, timber harvesting, and forest ecological services. Traditionally, operations on forest growing stock volume using field measurements are labor-intensive and time-consuming. Recently, remote sensing technology has emerged as a time-cost efficient method for forest inventory. In the present study, we have adopted three procedures, including samples expanding, feature selection, and results generation and evaluation. Extrapolating the samples from Light Detection and Ranging (LiDAR) scanning is the most important step in satisfying the requirement of sample size for nonparametric methods operation and result in accuracy improvement. Besides, mean decrease Gini (MDG) methodology embedded into Random Forest (RF) algorithm served as a selector for feature measure; afterwards, RF and K-Nearest Neighbor (KNN) were adopted in subsequent forest volume prediction. The results show that the retrieval of Forest volume in the entire area was in the range of 50−360 m3/ha, and the results from the two models show a better consistency while using the sample combination extrapolated by the optimal threshold value (2 × 10−4), leading to the best performances of RF (R2 = 0.618, root mean square error, RMSE = 43.641 m3/ha, mean absolute error, MAE = 33.016 m3/ha), followed by KNN (R2 = 0.617, RMSE = 43.693 m3/ha, MAE = 32.534 m3/ha). The detailed analysis that is discussed in the present paper clearly shows that expanding image-derived LiDAR samples helps in refining the prediction of regional forest volume while using satellite data and nonparametric models.

Fires are frequent in boreal forests affecting forest areas. The detection of forest disturbances and the monitoring of forest restoration are critical for forest management. Vegetation phenology information in remote sensing images may interfere with the monitoring of vegetation restoration, but little research has been done on this issue. Remote sensing and the geographic information system (GIS) have emerged as important tools in providing valuable information about vegetation phenology. Based on the MODIS and Landsat time-series images acquired from 2000 to 2018, this study uses the spatio-temporal data fusion method to construct reflectance images of vegetation with a relatively consistent growth period to study the vegetation restoration after the Greater Hinggan Mountain forest fire in the year 1987. The influence of phenology on vegetation monitoring was analyzed through three aspects: band characteristics, normalized difference vegetation index (NDVI) and disturbance index (DI) values. The comparison of the band characteristics shows that in the blue band and the red band, the average reflectance values of the study area after eliminating phenological influence is lower than that without eliminating the phenological influence in each year. In the infrared band, the average reflectance value after eliminating the influence of phenology is greater than the value with phenological influence in almost every year. In the second shortwave infrared band, the average reflectance value without phenological influence is lower than that with phenological influence in almost every year. The analysis results of NDVI and DI values in the study area of each year show that the NDVI and DI curves vary considerably without eliminating the phenological influence, and there is no obvious trend. After eliminating the phenological influence, the changing trend of the NDVI and DI values in each year is more stable and shows that the forest in the region was impacted by other factors in some years and also the recovery trend. The results show that the spatio-temporal data fusion approach used in this study can eliminate vegetation phenology effectively and the elimination of the phenology impact provides more reliable information about changes in vegetation regions affected by the forest fires. The results will be useful as a reference for future monitoring and management of forest resources.

The heat flux density of solar radiation, received by each surface of a double-block ballastless track bed slab, is closely related to its alignment and geographical latitude. In this work, a temperature field analysis model based on experimental data, the theories of solar radiation, and boundary heat transfer is established by a CRTS-I double-block ballastless track structure using the ABAQUS finite element software to investigate the influence of different alignments and geographical latitudes of the temperature field. The horizontal and vertical temperature gradients of the ballast bed plate were found to be in the most adverse conditions when the angle α n between the normal direction of the ballastless track slab bedside surface and positive south direction was equal to 90°. The standard deviation of the overall temperature gradient of the ballast bed was found to be at lowest and standard value of the dispersion degree was highest at an α n of 90°: 14.138 and 10.446°C/m, respectively. The horizontal and vertical temperature gradients in high latitudes and coastal areas were found to be more detrimental than that in the low latitudes or inland areas. These results can provide references for how to avoid high-temperature loading during railway line selection and track design.

Transcription factor RUNX1 holds an integral role in multiple-lineage haematopoiesis and is implicated as a cofactor in V(D)J rearrangements during lymphocyte development. Runx1 deficiencies resulted in immaturity and reduction of lymphocytes in mice. In this study, we found that runx1W84X/W84X mutation led to the reduction and disordering of B cells, as well as the failure of V(D)J rearrangements in B cells but not T cells, resulting in antibody-inadequate-mediated immunodeficiency in adult zebrafish. By contrast, T cell development was not affected. The decreased number of B cells mainly results from excessive apoptosis in immature B cells. Disrupted B cell development results in runx1W84X/W84X mutants displaying a similar phenotype to common variable immunodeficiency—a primary immunodeficiency disease primarily characterized by frequent susceptibility to infection and deficient immune response, with marked reduction of antibody production of IgG, IgA and/or IgM. Our studies demonstrated an evolutionarily conserved function of runx1 in maturation and differentiation of B cells in adult zebrafish, which will serve as a valuable model for the study of immune deficiency diseases and their treatments.

Liquid–liquid extraction (LLE) is the most commonly sample preparation procedure used by forensic toxicologists in China for screening drugs in whole human blood. It extracts numerous substances from blood including targeted drugs and interfering substances, specifically triglycerides (TG). With increasing prevalence of hyperlipidemia, the influences of TG on LLE and on subsequent analysis with gas chromatography–mass spectrometry (GC–MS) may become a major issue for forensic laboratories. This study aims to elucidate the influences of TG on LLE and to provide possible solutions to this problem. Nineteen commonly encountered drugs in forensic cases were spiked to human whole blood with different TG concentrations. Diethyl ether, ethyl acetate/hexane mixed solutions, chlorobutane and several other frequently used solvents were tested for the extraction of drugs from spiked whole blood. The supernatant organic layer was evaporated to dryness and reconstituted with methanol. The resultant products were analyzed by GC–MS, and the extraction recovery was calculated. LLE with diethyl ether, ethyl acetate/hexane (9:1) and chlorobutane all possessed effective and reliable extraction recoveries for blood sample with low TG concentrations (0.63–6.85 mmol/L). At high TG concentrations, diethyl ether produced a highly turbid substance that could not be further analyzed using GC–MS. Extraction recoveries drastically dropped for ethyl acetate/hexane (9:1) mixture at high TG concentrations, while chlorobutane experienced minimal drops in extraction recoveries. In conclusion, TG levels in whole blood noticeably influence drug recovery to variable extents depending on the LLE solvent. Chlorobutane showed minimal influences from TG content in whole blood and thus is the recommended LLE solvent for forensic drug extraction.

Abstract Background: Although a proportion of CSX patients have impaired brachial artery flow-mediated dilatation (FMD) in response to hyperemia, suggesting that endothelial dysfunction in these patients may be systemic and not just confined to the coronary circulation; the underlying mechanisms triggering endothelial dysfunction in these patients are still incompletely understood. Objectives: To assess the association of the index of Microcirculatory Resistance (IMR) with endothelial dysfunction and inflammation in patients with CSX. Methods: We studied 20 CSX patients and 20 age and gender-matched control subjects. Thermodilution-derived coronary flow reserve (CFR) and IMR were measured using a pressure-temperature sensor-tipped guidewire. Brachial artery FMD was measured using high-resolution, two-dimensional ultrasound images obtained with a Doppler ultrasound device (HDI-ATL 5000, USA) with a 5 MHz to 12 MHz linear-array transducer. Results: Compared with in control subjects, CFR was significantly lower (2.42 ± 0.78 vs. 3.59 ± 0.79, p < 0.001); IMR was higher (32.2 ± 8.0 vs. 19.5 ± 5.5, p < 0.001); the concentration of hs-CRP and FMD was higher (4.75 ± 1.62 vs. 2.75 ± 1.50; 5.24 ± 2.41 vs. 8.57 ± 2.46, p < 0.001) in CSX patients. The Duke treadmill score (DTS) was correlated positively to CFR and FMD (0.489 and 0.661, p < 0.001), it was negative to IMR and hsCRP (-0.761 and -0.087, p < 0.001) in CSX patients. Conclusions: The main finding in this study is that the DTS measured in patients with CSX was associated to hsCRP and FMD. Moreover, the independent effects of exercise tolerance can significantly impair FMD and hsCRP in CSX patients; especially it is particularly important to whom where FMD was associated negatively with IMR.

Neutrophils play important roles in innate immunity and are mainly dependent on various enzyme-containing granules to kill engulfed microorganisms. Zebrafish nephrosin (npsn) is specifically expressed in neutrophils; however, its function is largely unknown. Here, we generated an npsn mutant (npsnsmu5) via CRISPR/Cas9 to investigate the in vivo function of Npsn. The overall development and number of neutrophils remained unchanged in npsn-deficient mutants, whereas neutrophil antibacterial function was defective. Upon infection with Escherichia coli, the npsnsmu5 mutants exhibited a lower survival rate and more severe bacterial burden, as well as augmented inflammatory response to challenge with infection when compared with wild-type embryos, whereas npsn-overexpressing zebrafish exhibited enhanced host defence against E. coli infection. These findings demonstrated that zebrafish Npsn promotes host defence against bacterial infection. Furthermore, our findings suggested that npsn-deficient and -overexpressing zebrafish might serve as effective models of in vivo innate immunity.

Glucose is a main carbon and energy source for virus proliferation and is usually involved in the glycolysis, pentose phosphate pathway (PPP), and tricarboxylic acid cycle (TCA cycle) pathways. In this study, we investigated the roles of glucose-related metabolic pathways during the replication of infectious spleen and kidney necrosis virus (ISKNV), which has caused serious economic losses in the cultured Chinese perch (Siniperca chuatsi) industry. We found that ISKNV infection enhanced the metabolic pathways of the PPP and the TCA cycle at the early stage of the ISKNV infection cycle and enhanced the glycolysis pathway at the late stage of the ISKNV infection cycle though the comprehensive analysis of transcriptomics, proteomics, and metabolomics. The advanced results proved that ISKNV replication induced upregulation of aerobic glycolysis at the late stage of ISKNV infection cycle and aerobic glycolysis were required for ISKNV multiplication. In addition, the PPP, providing nucleotide biosynthesis, was also required for ISKNV multiplication. However, the TCA cycle involving glucose was not important and necessary for ISKNV multiplication. The results reported here provide new insights into viral pathogenesis mechanism of metabolic shift, as well as antiviral treatment strategies.

A series of 5-arylisoxazole derivatives were synthesized via the reaction of 3-(dimethyl-amino)-1-arylprop-2-en-1-ones with hydroxylamine hydrochloride in aqueous media without using any catalyst. This method has the advantages of easier work-up, mild reaction conditions, high yields, and an environmentally benign procedure.