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Background: Autism spectrum disorder (ASD) is a multifactorial, pervasive, neurodevelopmental disorder, of which intestinal symptoms collectively represent one of the most common comorbidities. Methods: In this study, 1,222 children with ASD and 1,206 typically developing (TD) children aged 2-7 years were enrolled from 13 cities in China. Physical measurement and basic information questionnaires were conducted in ASD and TD children. The Childhood Autism Rating Scale (CARS), Social Responsiveness Scale (SRS), and Autism Behavior Checklist (ABC) were used to evaluate the clinical symptoms of children with ASD. The six-item Gastrointestinal Severity Index (6-GSI) was used to evaluate the prevalence of intestinal symptoms in two groups. Results: The detection rates of constipation, stool odor, and total intestinal symptoms in ASD children were significantly higher than those in TD children (40.098% vs. 25.622%, 17.021% vs. 9.287%, and 53.601% vs. 41.294%, respectively). Autistic children presenting with intestinal comorbidity had significantly higher scores on the ABC, SRS, CARS, and multiple subscales than autistic children without intestinal symptoms, suggesting that intestinal comorbidity may exacerbates the core symptoms of ASD children. Conclusion: Intestinal dysfunction was significantly more common in autistic than in TD children. This dysfunction may aggravate the core symptoms of children with ASD.

The heart, which is the first organ to develop, is highly dependent on its form to function1,2. However, how diverse cardiac cell types spatially coordinate to create the complex morphological structures that are crucial for heart function remains unclear. Here we integrated single-cell RNA-sequencing with high-resolution multiplexed error-robust fluorescence in situ hybridization to resolve the identity of the cardiac cell types that develop the human heart. This approach also provided a spatial mapping of individual cells that enables illumination of their organization into cellular communities that form distinct cardiac structures. We discovered that many of these cardiac cell types further specified into subpopulations exclusive to specific communities, which support their specialization according to the cellular ecosystem and anatomical region. In particular, ventricular cardiomyocyte subpopulations displayed an unexpected complex laminar organization across the ventricular wall and formed, with other cell subpopulations, several cellular communities. Interrogating cell-cell interactions within these communities using in vivo conditional genetic mouse models and in vitro human pluripotent stem cell systems revealed multicellular signalling pathways that orchestrate the spatial organization of cardiac cell subpopulations during ventricular wall morphogenesis. These detailed findings into the cellular social interactions and specialization of cardiac cell types constructing and remodelling the human heart offer new insights into structural heart diseases and the engineering of complex multicellular tissues for human heart repair.

BackgroundPancreatic neuroendocrine tumors (PNETs) with low microvessel density and fibrosis often exhibit clinical aggressiveness. Given the contribution of cancer-associated fibroblasts (CAFs) to the hypovascular fibrotic stroma in pancreatic ductal adenocarcinoma, investigating whether CAFs play a similar role in PNETs becomes imperative. In this study, we investigated the involvement of CAFs in PNETs and their effects on clinical outcomes.MethodsWe examined 79 clinical PNET specimens to evaluate the number and spatial distribution of α-smooth muscle actin (SMA)-positive cells, which are indicative of CAFs. Then, the findings were correlated with clinical outcomes. In vitro and in vivo experiments were conducted to assess the effects of CAFs (isolated from clinical specimens) on PNET metastasis and growth. Additionally, the role of the stromal-cell-derived factor 1 (SDF1)-AGR2 axis in mediating communication between CAFs and PNET cells was investigated.ResultsαSMA-positive and platelet-derived growth factor-α-positive CAFs were detected in the hypovascular stroma of PNET specimens. A higher abundance of α-SMA-positive CAFs within the PNET stroma was significantly associated with a higher level of clinical aggressiveness. Notably, conditioned medium from PNET cells induced an inflammatory phenotype in isolated CAFs. These CAFs promoted PNET growth and metastasis. Mechanistically, PNET cells secreted interleukin-1, which induced the secretion of SDF1 from CAFs. This cascade subsequently elevated AGR2 expression in PNETs, thereby promoting tumor growth and metastasis. The downregulation of AGR2 in PNET cells effectively suppressed the CAF-mediated promotion of PNET growth and metastasis.ConclusionCAFs drive the growth and metastasis of aggressive PNETs. The CXCR4-SDF1 axis may be a target for antistromal therapy in the treatment of PNET. This study clarifies mechanisms underlying PNET aggressiveness and may guide future therapeutic interventions targeting the tumor microenvironment.

Cells interpret a variety of signals through G-protein-coupled receptors (GPCRs) and stimulate the generation of second messengers such as cyclic adenosine monophosphate (cAMP). A long-standing puzzle is deciphering how GPCRs elicit different physiological responses despite generating similar levels of cAMP. We previously showed that some GPCRs generate cAMP from both the plasma membrane and the Golgi apparatus. Here we demonstrate that cardiomyocytes distinguish between subcellular cAMP inputs to elicit different physiological outputs. We show that generating cAMP from the Golgi leads to the regulation of a specific protein kinase A (PKA) target that increases the rate of cardiomyocyte relaxation. In contrast, cAMP generation from the plasma membrane activates a different PKA target that increases contractile force. We further validated the physiological consequences of these observations in intact zebrafish and mice. Thus, we demonstrate that the same GPCR acting through the same second messenger regulates cardiac contraction and relaxation dependent on its subcellular location.

Abstract Objectives To explore the association between hyperuricemia and having periodontitis. Materials and methods A representative cross-sectional dataset of 10,158 adults was extracted from the National Health and Nutrition Examination Survey (NHANES) 2009–2014. The association between hyperuricemia (the primary exposure) and having periodontitis (outcome) were evaluated using weighted logistic regression models. Serum uric acid (UA) levels and the UA to creatinine (UA/Cr) ratio were used as secondary exposures. Their associations with the diagnosis periodontitis were analyzed using weighted logistic regression or restricted cubic spline regression. Results The prevalence of Stages III/IV periodontitis was 47.7% among individuals with hyperuricemia and 37.4% among those without. After adjustment, individuals with hyperuricemia had 0.281 times higher odds of developing Stages III/IV periodontitis compared to those without hyperuricemia (adjusted OR = 1.286, 95% CI = 1.040 to 1.591, P = 0.024). The increased odds could be explained by a linear relationship with the serum UA/Cr ratio and a U-shaped relationship with serum UA levels. Each unit increase in the serum UA/Cr ratio was associated with 0.048 times higher odds of developing Stages III/IV periodontitis (adjusted OR = 1.048, 95% CI = 1.008 to 1.088, P = 0.021). Additionally, each 1 mg/dL increase in serum UA was associated with 0.156 times higher odds (adjusted OR = 1.156, 95% CI = 1.009 to 1.323, P = 0.038) of developing Stages III/IV periodontitis when UA levels were greater than 5.9 mg/dL, but 0.118 times lower odds when UA levels were 5.9 mg/dL or lower (adjusted OR = 0.882, 95% CI = 0.790 to 0.984, P = 0.027). Sensitivity analyses validated the robustness of the findings. Conclusions This study provides the first direct evidence that hyperuricemia is associated with Stages III/IV periodontitis. Clinical relevance Hyperuricemia may represent a new potential comorbidity of periodontitis, possibly contributing directly or indirectly to the disease burden in patients with periodontitis. Clinical trial number Not applicable.

Abstract Background Lysosomal homeostasis and functions are essential for the survival of neural cells. Impaired lysosomal biogenesis and acidification in Alzheimer’s disease (AD) pathogenesis leads to proteolytic dysfunction and neurodegeneration. However, the key regulatory factors and mechanisms of lysosomal homeostasis in AD remain poorly understood. Methods ROCK1 expression and its co-localization with LAMP1 and SQSTM1/p62 were detected in post-mortem brains of healthy controls and AD patients. Lysosome-related fluorescence probe staining, transmission electron microscopy and immunoblotting were performed to evaluate the role of ROCK1 in lysosomal biogenesis and acidification in various neural cell types. The interaction between ROCK1 and TFEB was confirmed by surface plasmon resonance and in situ proximity ligation assay (PLA). Moreover, we performed AAV-mediated ROCK1 downregulation followed by immunofluorescence, enzyme-linked immunosorbent assay (ELISA) and behavioral tests to unveil the effects of the ROCK1–TFEB axis on lysosomes in APP/PS1 transgenic mice. Results ROCK1 level was significantly increased in the brains of AD individuals, and was positively correlated with lysosomal markers and Aβ. Lysosomal proteolysis was largely impaired by the high abundance of ROCK1, while ROCK1 knockdown mitigated the lysosomal dysfunction in neurons and microglia. Moreover, we verified ROCK1 as a previously unknown upstream kinase of TFEB independent of m-TOR or GSK-3β. ROCK1 elevation resulted in abundant extracellular Aβ deposition which in turn bound to Aβ receptors and activated RhoA/ROCK1, thus forming a vicious circle of AD pathogenesis. Genetically downregulating ROCK1 lowered its interference with TFEB, promoted TFEB nuclear distribution, lysosomal biogenesis and lysosome-mediated Aβ clearance, and eventually prevented pathological traits and cognitive deficits in APP/PS1 mice. Conclusion In summary, our results provide a mechanistic insight into the critical role of ROCK1 in lysosomal regulation and Aβ clearance in AD by acting as a novel upstream serine kinase of TFEB.

Abstract Introduction Post-keratoplasty glaucoma (PKG) is a major complication following penetrating keratoplasty (PKP) for congenital corneal opacity (CCO). This study aims to assess the preoperative structural risk factors for PKG following PKP for CCO using ultrasound biomicroscopy (UBM). Methods Pediatric patients with CCO who underwent preoperative UBM and primary PKP were enrolled. Patients with anterior segment operation history or with a follow-up duration less than 12 months were excluded. The structural features of the anterior segment including central corneal thickness, anterior chamber depth, angle closure range (ACR), anterior synechia range, maximum iridocorneal adhesion length, abnormal iridocorneal synechia, and lens anomalies were identified on UBM images. The medical histories were reviewed to identify clinical features. The incidence of PKG was assessed to determine significant structural and clinical risk factors. Results Fifty-one eyes of 51 pediatric patients with CCO were included. The median age at surgery was 8.0 months, and the mean follow-up duration was 33 ± 9 months. Eleven (21.6%) eyes developed PKG. The main structural risk factors were abnormal iridocorneal synechia (P = 0.015), lens anomaly (P = 0.001), and larger ACR (P = 0.045). However, a larger range of normal anterior synechia without involvement of the angle was not a significant risk factor. Preoperative glaucoma (P

Abstract Blowouts are a common type of wind-eroded landform found in sandy and desertified areas. They also represent a major degradative surface process affecting grassland ecosystems. Blowouts exacerbate changes in surface morphology through their effects on other surface phenomena including vegetation. In this paper, Xilingol League sandy grassland blowouts are taken as the research object, and the U.S. Keyhole satellite data and China’s Gaofen-1 satellite data are used as the data source, and the blowouts are extracted based on the 3 S technology for a total of six periods of high-resolution remote sensing image data in the study area from 1962 to 2023. The Landscape Pattern Index method and Fuzzy Land Use Simulation (FLUS) modelling applied to changes over the last six decades provided spatial evolution parameters for predicting future blowout distributions. Results showed that blowouts affecting the Xilingol grassland area increased by 16.81% over the past 60 years. The patch density (PD) increased by 0.9 per hectare. The mean proximity index (PROX_MN) and mean Euclidean nearest neighbour distance (ENN_MN) showed a tendency to decrease and then increase indicating initial expansion and then merging of adjacent blowouts to create the present landscape. The FLUS model used ten factors to predict changes in blowout distributions from 2023 to 2033. Factors included digital elevation model (DEM), slope, aspect, normalized difference vegetation index (NDVI), mean annual temperature, mean annual precipitation, population density, real GDP, distance to water, and distance to impervious surfaces. It was found that grassland area decreased by 6217.12 hm2 and blowout area decreased by 102.91 hm2. Results of this study can expand understanding of blowout morphodynamics in ecologically sensitive areas.

In this contribution, the complicated dynamical behaviors and optimal harvesting policy of a discrete-time predator–prey model with fear and refuge effects are formulated. Both the fear and prey refuge effects refer to an interaction between predator and prey. In the first place, the existence and local stability of three fixed points of proposed model are investigated by virtue of our methodology, that is, the eigenvalues of the Jacobian matrix. One step further, it is worth mentioning that the model undergoes flip bifurcation (i.e., period–doubling bifurcation) and Neimark–Sacker bifurcation at the interior fixed point by the utilization of bifurcation theory and center manifold theory. Also, optimal harvesting strategy is investigated, and the expressions of optimal harvesting efforts are determined. Two examples, in the end, are put forward to prove that they are consistent with the previous theoretical results.

Abstract Adeno-associated viruses (AAVs) are foundational gene delivery tools for basic science and clinical therapeutics. However, lack of mechanistic insight, especially for engineered vectors created by directed evolution, can hamper their application. Here, we adapt an unbiased human cell microarray platform to determine the extracellular and cell surface interactomes of natural and engineered AAVs. We identify a naturally-evolved and serotype-specific interaction between the AAV9 capsid and human interleukin 3 (IL3), with possible roles in host immune modulation, as well as lab-evolved low-density lipoprotein receptor-related protein 6 (LRP6) interactions specific to engineered capsids with enhanced blood-brain barrier crossing in non-human primates after intravenous administration. The unbiased cell microarray screening approach also allows us to identify off-target tissue binding interactions of engineered brain-enriched AAV capsids that may inform vectors’ peripheral organ tropism and side effects. Our cryo-electron tomography and AlphaFold modeling of capsid-interactor complexes reveal LRP6 and IL3 binding sites. These results allow confident application of engineered AAVs in diverse organisms and unlock future target-informed engineering of improved viral and non-viral vectors for non-invasive therapeutic delivery to the brain.

Research has revealed the positive impact of intelligent personal assistants (IPAs) on L2 learners' oral development and learning attitude. These studies, however, focused mostly on the in-class use of IPAs, with existing research on the out-of-class use being exploratory. To fill the gap of lacking empirical investigations on IPA-based autonomous second language learning (ASLL), this study recruited 34 college EFL learners to use Google Assistant (GA) in their respective homes for six weeks and examined their listening and speaking development afterwards. Each week, participants received an email about the new commands for them to explore, and half of them were additionally given sustained teacher guidance (i.e. weekly evaluation forms) on the usefulness of each command to better enhance their participation. Statistical analysis on the participants' learning gains demonstrated that participants receiving weekly evaluation forms made significant improvement in both listening and speaking skills, and they even outperformed those without weekly evaluation forms in their oral development. Nevertheless, there was no significant difference in the two groups' listening gains. These findings demonstrate that, with sustained teacher guidance, IPAs have great potential in L2 oral development in ASLL context, but more long-term studies on IPAs' impact for L2 listening should be conducted.

Purpose: This study aims to evaluate the current situation of education for sustainable development, climate change education and environmental education in a nationwide context. Methodologically, this study calls for more research to go beyond case studies and take a similar approach to examine university curricula and facilitate cross-country comparisons. Design/methodology/approach: This paper examines the status of climate and sustainability curricula in Taiwan's higher education system. Using the course catalog for the 2020-2021 academic year, the authors constructed a unique data set that includes 1,827 courses at 29 major universities in Taiwan. In each institution, the authors search for course titles that include "climate," "sustainable/sustainability" and "environment/environmental" as keywords and code the courses according to their disciplines. Findings: The finding highlights the variations across institutional types and subject matters. Public universities have an average of 4.94 related courses per 1,000 students, whereas private universities have only 3.13. In general, the relevant courses are more concentrated in the STEM and bioscience fields. The curricula, however, are seriously constrained by the disciplinary structure and foster few transdisciplinary perspectives. Originality/value: The authors seek to go beyond case studies and offer one of the most comprehensive curricula samples at the national level. Taiwan adds an important data point, as the current literature focuses heavily on the USA and Europe.

The quantification of protein biomarkers in blood at picomolar-level sensitivity requires labour-intensive incubation and washing steps. Sensing proteins in sweat, which would allow for point-of-care monitoring, is hindered by the typically large interpersonal and intrapersonal variations in its composition. Here we report the design and performance of a wearable and wireless patch for the real-time electrochemical detection of the inflammatory biomarker C-reactive (CRP) protein in sweat. The device integrates iontophoretic sweat extraction, microfluidic channels for sweat sampling and for reagent routing and replacement, and a graphene-based sensor array for quantifying CRP (via an electrode functionalized with anti-CRP capture antibodies-conjugated gold nanoparticles), ionic strength, pH and temperature for the real-time calibration of the CRP sensor. In patients with chronic obstructive pulmonary disease, with active or past infections or who had heart failure, the elevated concentrations of CRP measured via the patch correlated well with the protein's levels in serum. Wearable biosensors for the real-time sensitive analysis of inflammatory proteins in sweat may facilitate the management of chronic diseases.