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Abstract Breast cancer is one of the most threatening women health diseases worldwide and its molecular heterogeneity offers a range of response to therapy. The role of polyamine metabolism is receiving increasing attention. Polyamine metabolism not only plays an important role in the occurrence and development of breast cancer, but also interacts with tumor immune microenvironment. In this work, we applied single-cell RNA-sequencing (scRNA-seq) and systems immunological approaches to interrogate immune cell infiltration gene-to-gene co-expressions in the bulk tumor transcriptomes of breast cancer. We acquired breast cancer sample data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO), evaluated the infiltration status of 22 immune cell types using CIBERSORTx tool, respectively. By leveraging the Retrospective Breast sample of various technologies including gene expression and methylation, we identified 46 breast cancer proliferation-associated co-expression modules using weighted gene coexpression network analysis (WGCNA) approach along with machine learning models which in turn delineated single cell level expressions features that these selected module possessed. We observed substantial cellular heterogeneity in the breast cancer microenvironment, where lineage-specific gene expression patterns were highly associated with tumor progression. Moreover, we also identified the gene modules correlated with immune cell infiltration level that could function as regulators in response to tumors for immune therapy. Moreover, risk scores were correlated with immune cell function in different patient groups defined by high- and low-risk. The findings of this study shed a new light upon molecular classification prognostic assessment and personalized treatment in breast cancer.

Abstract Current gene therapy for Duchenne muscular dystrophy (DMD) utilizes adeno-associated virus (AAV) to deliver micro-dystrophin (µDys), which does not provide full protection for striated muscles as it lacks many important functional domains of full-length (FL) dystrophin. Here we develop a triple vector system to deliver FL-dystrophin into skeletal and cardiac muscles. We split FL-dystrophin into three fragments linked to two orthogonal pairs of split intein, allowing efficient assembly of FL-dystrophin. The three fragments packaged in myotropic AAV (MyoAAV4A) restore FL-dystrophin expression in both skeletal and cardiac muscles in male mdx 4cv mice. Dystrophin-glycoprotein complex components are also restored at the sarcolemma of dystrophic muscles. MyoAAV4A-delivered FL-dystrophin significantly improves muscle histopathology, contractility, and overall strength comparable to µDys, but unlike µDys, it also restores defective cavin 4 localization and associated signaling in mdx 4cv heart. Therefore, our data support the feasibility of a mutation-independent FL-dystrophin gene therapy for DMD, warranting further clinical development.

Introduction: Prophylactic antifungal therapy has been widely used for critical patients, but it has failed to improve patient prognosis and has become a hot topic. This may be related to disruption of fungal homeostasis, but the mechanism of fungi action is not clear. As a common pathway in critical patients, intestinal ischemia–reperfusion (IIR) injury is fatal and regulated by gut microbiota. However, the exact role of enteric fungi in IIR injury remains unclear. Objectives: This is a clinical study that aims to provide new perspectives in clarifying the underlying mechanism of IIR injury and propose potential strategies that could be relevant for the prevention and treatment of IIR injury in the near future. Methods: ITS sequencing was performed to detect the changes in fungi before and after IIR injury. The composition of enteric fungi was altered by pretreatment with single-fungal strains, fluconazole and mannan, respectively. Intestinal morphology and function impairment were evaluated in the IIR injury mouse model. Intestinal epithelial MODE-K cells and macrophage RAW264.7 cells were cultured for in vitro tests. Results: Fecal fungi diversity revealed the obvious alteration in IIR patients and mice, accompanied by intestinal epithelial barrier dysfunction. Fungal colonization and mannan supplementation could reverse intestinal morphology and function impairment that were exacerbated by fluconazole via inhibiting the expression of SAA1 from macrophages and decreasing pyroptosis of intestinal epithelial cells. Clodronate liposomes were used to deplete the number of macrophages, and it was demonstrated that the protective effect of mannan was dependent on macrophage involvement. Conclusion: This finding firstly validates that enteric fungi play a crucial role in IIR injury. Preventive antifungal treatment should consider damaging fungal balance. This study provides a novel clue to clarify the role of enteric fungi in maintaining intestinal homeostasis.

Acetaminophen (APAP) overdose is a leading cause of drug-induced liver injury (DILI), with gender-specific differences in susceptibility. However, the mechanism underlying this phenomenon remains unclear. Our study reveals that the gender-specific differences in susceptibility to APAP-induced hepatotoxicity are due to differences in the gut microbiota. Through microbial multi-omics and cultivation, we observed increased gut microbiota-derived deguelin content in both women and female mice. Administration of deguelin was capable of alleviating hepatotoxicity in APAP-treated male mice, and this protective effect was associated with the inhibition of hepatocyte oxidative stress. Mechanistically, deguelin reduced the expression of thyrotropin receptor (TSHR) in hepatocytes with APAP treatment through direct interaction. Pharmacologic suppression of TSHR expression using ML224 significantly increased hepatic glutathione (GSH) in APAP-treated male mice. These findings suggest that gut microbiota-derived deguelin plays a crucial role in reducing APAP-induced hepatotoxicity in female mice, offering new insights into therapeutic strategies for DILI.

Abstract Background Previous studies have found that frailty and sarcopenia are commonly diagnosed in inflammatory bowel disease (IBD) patients, indicating an association between these conditions. Nonetheless, the cause‒effect connection between IBD, frailty, and sarcopenia remains unclear. Methods We sourced the genetic variants for the exposures and outcomes from publicly accessible, extensive genome-wide association studies (GWAS). Specifically, we obtained IBD data from the International IBD Genetics Consortium, frailty index (FI) data from the United Kingdom Biobank and Swedish TwinGene, and sarcopenia data from a recent GWAS meta-analysis. Five methods, including inverse variance weighted (IVW), simple mode, MR-Egger, weighted mode, and the weighted median, were used to proceed with MR estimates. We also performed heterogeneity and horizontal pleiotropy tests. Results Our results indicated a positive causal relationship between ulcerative colitis (UC) (IVW: β = 0.014, 95% CI, 0.006 to 0.021, p = 0.001) and Crohn's disease (CD) (IVW: β = 0.012; 95% CI, 0.006 to 0.018, p = 2e−04) with the FI. However, we uncovered no proof of a cause-and-effect relationship between UC (IVW: β = 0.001, 95% CI, −0.015 to 0.017, p = 0.344) or CD (IVW: β = 0.003, 95% CI, −0.009 to 0.015, p = 0.214) and sarcopenia. Additionally, in the inverse order, we also discovered no cause-and-effect connection between FI or sarcopenia on UC or CD in this study. Conclusion The MR analysis showed a positive causal association between IBD and FI, indicating that IBD patients may exhibit aging-related characteristics. Therefore, frailty assessments should be conducted as early as possible in IBD patients.

Abstract Background Inflammatory Bowel Diseases (IBD), an autoimmune disease characterised by abnormal intestinal immunity, are related to vital morbidity around the world. However, therapeutic agents for IBD have not achieved desired benefit. Exploring new therapeutic targets for IBD, especially based on its abnormally intestinal immunity, could alleviate the flare-up and worsening of IBD. Tissue resident memory T cells (TRM) are core of multiple autoimmune diseases, including IBD. However, the mechanism of TRM differentiation remains to be investigated. Methods The alterations in mRNA and lncRNA profile of intestinal intraepithelial lymphocytes (IELs), the largest component of intestinal TRM, were analyzed in DSS-induced chronic colitis. Based on it, we examined the function of rectal insulin instillation in a dextran sodium sulfate (DSS) induced chronic colitis. Furthermore, we investigated the downstream-target of the insulin pathway—EZH2 and the crucial role of EZH2 in intestinal tissue resident memory T cell differentiation by utilizing EZH2fl/flCD4cre mice. Results Insulin receptor (INSR) expression was found to be significantly reduced. Activation of mucosal insulin pathway by rectal insulin instillation exacerbated colitis by disrupting IELs subgroups and up-regulating TNF-ɑ and IL-17 expression. Rectal insulin instillation promoted EZH2 expression and EZH2 inhibition alleviated chronic colitis. EZH2fl/flCD4cre mice restored the normal IEL subgroups and suppressed TNF-ɑ and IL-17 expression, exhibiting alleviated colitis. IELs from EZH2fl/flCD4cre mice exhibit significant changes in TRM related phenotype. CD4+TRM was significantly increased in chronic colitis and decreased in EZH2fl/flCD4cre mice. Conclusion Insulin receptor of intestinal mucosal T-cells could promote intestinal TRM differentiation via EZH2. Our discoveries suggest that therapies targeting colonic INSR and EZH2 could be potential treatment for IBD based on its regulatory effects on TRM. Insulin receptor inhibitors rather than insulin should be applied during colitis-active phase. In addition, EZH2 shows to be a downstream signal of the insulin pathway and EZH2 inhibitor could alleviating intestinal inflammation. However, the critical role of EZH2 in TRM differentiation restricts the anti-tumor effects of EZH2 inhibitor in vivo.

Knowledge graph embedding represents entities and relations as low-dimensional continuous vectors. Recently, researchers have attempted to leverage the potential semantic connections between entities with hierarchical relationships in the knowledge graph. However, existing knowledge embedding methods that model entity hierarchical structures face the following challenges: (1) Setting the same embedding dimension for entities at different hierarchical levels can lead to overfitting issues and the waste of storage and computational resources; (2) Manually searching the embedding dimension in discrete space makes it easy to miss the optimal parameters and increases training costs. Therefore, we propose a knowledge embedding method, HEAES, that automatically learns the embedding dimensions for entities based on their hierarchical structure. Specifically, we first propose a new modeling method to capture the hierarchical relationships of entities, and we then train the model on a triple classification task. Then, we adaptively learn the pruning threshold to trim the embedding vectors, automatically learning different embedding dimensions for entities at different hierarchical levels. Experiments on the YAGO26K and DB111K datasets verify that introducing entity hierarchical information helps boost the model performance.

Background: The diagnosis and evaluation of the severity of acute mountain sickness (AMS) continue to be problematic due to a lack of consensus on the inclusion of symptoms in a scoring system. Recent investigations highlight the significance of gastrointestinal symptoms in identifying this condition. However, the specific gastrointestinal symptoms associated with AMS have not been thoroughly elucidated in previous studies, and the underlying risk factors remain inadequately comprehended. Methods: This study aimed to investigate the characteristics, trends, and risk factors related to gastrointestinal symptoms encountered during train travel to high altitude. A total of 69 passengers, specifically all with medical backgrounds, were surveyed 6 times over a period of 14 days. Results: The daily incidence of abdominal discomfort was higher than non-gastrointestinal symptoms within 14 days. Gastrointestinal symptoms demonstrated a greater prevalence, longer duration, and increased risk compared to non-gastrointestinal symptoms, such as headaches. The symptoms of abdominal distension and bowel sound hyperaction were found to be prevalent and persistent among patients diagnosed with AMS, exhibiting a high incidence rate. Gender, age, body mass index (BMI), smoking habits, and alcohol consumption were identified as risk factors associated with the occurrence and duration of gastrointestinal symptoms. Conclusion: This study suggests that gastrointestinal symptoms are more common and persistent when traveling to the plateau by train. These symptoms should be taken into consideration in the further diagnosis and prevention of AMS. Therefore, this study provides a significant theoretical foundation for the prevention and treatment of AMS.

Abstract Lipid metabolic reprogramming is closely related to tumor progression with the mechanism not fully elucidated. Here, we report the immune‐regulated role of lanosterol synthase (LSS), an essential enzyme in cholesterol synthesis. Database analysis and clinical sample experiments suggest that LSS was lowly expressed in colon and breast cancer tissues, which indicates poor prognosis. The biological activity of tumor cell lines and tumor progression in NOD scid gamma (NSG) mice were not affected after LSS knockdown, whereas LSS deficiency obviously aggravated tumor burden in fully immunized mice. Flow cytometry analysis showed that LSS knockdown significantly promoted the formation of tumor immunosuppressive microenvironment, characterized by the increase in M2 macrophages and polymorphonuclear myeloid‐derived suppressor cells (PMN‐MDSCs), as well as the decrease in anti‐tumoral T lymphocytes. With the inhibition of myeloid infiltration or loss function of T lymphocytes, the propulsive effect of LSS knockdown on tumor progression disappeared. Mechanistically, LSS knockdown increased programmed death ligand 1 (PDL1) protein stability by 2,3‐oxidosqualene (OS) binding to PDL1 protein. Anti‐PDL1 therapy abolished LSS deficiency‐induced immunosuppressive microenvironment and cancer progression. In conclusion, our results show that LSS deficiency promotes tumor progression by establishing an OS–PDL1 axis‐dependent immunosuppressive microenvironment, indicative of LSS or OS as a potential hallmark of response to immune checkpoint blockade.

The unique palindromic inverted terminal repeats (ITRs) and single-stranded nature of adeno-associated virus (AAV) DNA are major hurdles to current sequencing technologies. Due to these characteristics, sequencing noncanonical AAV genomes present in AAV vector preparations remains challenging. To address this limitation, we developed thorough molecule configuration analysis of noncanonical AAV genomes (TMCA-AAV-seq). TMCA-AAV-seq takes advantage of the documented AAV packaging mechanism in which encapsidation initiates from its 3′ ITR, for AAV-seq library construction. Any AAV genome with a 3′ ITR is converted to a template suitable to adapter addition by a Bst DNA polymerase-mediated extension reaction. This extension reaction helps fix ITR heterogeneity in the AAV population and allows efficient adapter addition to even noncanonical AAV genomes. The resulting library maintains the original AAV genome configurations without introducing undesired changes. Subsequently, long-read sequencing can be performed by the Pacific Biosciences (PacBio) single-molecule, real-time (SMRT) sequencing technology platform. Finally, through comprehensive data analysis, we can recover canonical, noncanonical AAV DNA, and non-AAV vector DNA sequences, along with their molecular configurations. Our method is a robust tool for profiling thorough AAV-population genomes. TMCA-AAVseq can be further extended to all parvoviruses and their derivative vectors.

Self-complementary AAV vectors (scAAV) use a mutant inverted terminal repeat (mITR) for efficient packaging of complementary stranded DNA, enabling rapid transgene expression. However, inefficient resolution at the mITR leads to the packaging of monomeric or subgenomic AAV genomes. These noncanonical particles reduce transgene expression and may affect the safety of gene transfer. To address these issues, we have developed a novel class of scAAV vectors called covalently closed-end double-stranded AAV (cceAAV) that eliminate the mITR resolution step during production. Instead of using a mutant ITR, we used a 56-bp recognition sequence of protelomerase (TelN) to covalently join the top and bottom strands, allowing the vector to be generated with just a single ITR. To produce cceAAV vectors, the vector plasmid is initially digested with TelN, purified, and then subjected to a standard triple-plasmid transfection protocol followed by traditional AAV vector purification procedures. Such cceAAV vectors demonstrate yields comparable to scAAV vectors. Notably, we observed enhanced transgene expression as compared to traditional scAAV vectors. The treatment of mice with hemophilia B with cceAAV-FIX resulted in significantly enhanced long-term FIX expression. The cceAAV vectors hold several advantages over scAAV vectors, potentially leading to the development of improved human gene therapy drugs.

Abstract Damage to the intestinal epithelial barrier (IEB) has been reported under high-altitude (HA) conditions and may be responsible for HA-associated gastrointestinal (GI) disorders. However, this pathogenetic mechanism does not fully explain the GI stress symptoms, such as flatulence and motility diarrhea, which accompany the IEB damage under HA conditions, especially for the people exposed to HA acutely. In the present study, we collected the blood samples from the people who lived at HA and found the concentration of enteric glial cells (EGCs)-associated biomarkers increased significantly. HA mouse model was then established and the results revealed that EGCs were involved in IEB damage. Zona occludens (ZO)-1, occludin, and claudin-1 expression was negatively correlated with that of glial fibrillary acidic protein (GFAP) and S100β under HA conditions. In order to learn more about how EGCs influence IEB, the in vitro EGC and MODE-K hypoxia experiments that used hypoxic stimulation for simulating in vivo exposure to HA was performed. We found that hypoxia increased S100β secretion in EGCs. And MODE-K cells cultured in medium conditioned by hypoxic EGCs showed low ZO-1, occludin, and claudin-1 levels of expression. Furthermore, treatment of MODE-K cells with recombinant mouse S100β resulted in diminished levels of ZO-1, occludin, and claudin-1 expression. Thus, HA exposure induces greater S100β secretion by EGCs, which aggravates the damage to the IEB. This study has revealed a novel mechanism of IEB damage under HA conditions, and suggest that EGCs may constitute a fresh avenue for the avoidance of GI disorders at HA.

Abstract Background Gene editing has emerged as an exciting therapeutic development platform for numerous genetic and nongenetic diseases. Targeting lipid-modulating genes such as angiopoietin-related protein 3 (ANGPTL3) with gene editing offers hope for a permanent solution to lower cardiovascular disease risks associated with hypercholesterolemia. Results In this study, we developed a hepatocyte-specific base editing therapeutic approach delivered by dual adeno-associated virus (AAV) to enable hepatocyte-specific targeting of Angptl3 to lower blood lipid levels. Systemic AAV9-mediated delivery of AncBE4max, a cytosine base editor (CBE), targeting mouse Angptl3 resulted in the installation of a premature stop codon in Angptl3 with an average efficiency of 63.3 ± 2.3% in the bulk liver tissue. A near-complete knockout of the ANGPTL3 protein in the circulation were observed within 2–4 weeks following AAV administration. Furthermore, the serum levels of triglyceride (TG) and total cholesterol (TC) were decreased by approximately 58% and 61%, respectively, at 4 weeks after treatment. Conclusions These results highlight the promise of liver-targeted Angptl3 base editing for blood lipid control.

With the rapid development of urbanization, the disorderly construction land expansion takes up too much ecological space, leading to the exacerbation of the regional internal ecosystem. Therefore, exploring the coordinated relationship between construction land expansion and ecological environment change has become a key issue for regional sustainable development. Based on remote sensing technology, the evaluation system of construction land expansion and ecological environment change in the Three Gorges Reservoir area was constructed. The spatial–temporal evolution of construction land expansion and ecological environment change from 1995 to 2020 and their coordination were analyzed, and they were divided into coupling and coordinated development types. The research shows the following findings: Firstly, in the year 2020, the comprehensive value of construction land expansion reached 0.8635, an increase of more than 8 times compared to 1995, indicating a significant trend of rapid expansion. Secondly, the comprehensive value of ecological environment change from 1995 to 2020 remained between 0.4 and 0.6 and continued to change in a good direction. And improving the remote sensing ecological indicators has expanded the difference in ecological environment quality in the Three Gorges Reservoir Area. Thirdly, the coupled model increased from 0.0534 to 0.7951 from 1995 to 2020, which is in a low-level and slow-growing state, and social and economic development does not completely depend on destroying the ecological environment quality. Finally, three types of coordinated development are divided: ecological environment protection type, development and protection in tandem type, and ecological environment consumable type. Differentiated regulation strategies and suggestions are also proposed.

Genetic engineering advances have led to recombinant adeno-associated virus (rAAV) becoming an invaluable tool for the development of effective gene therapies. The production of rAAV is susceptible to off-target heterogeneous packaging, the effects of which are still being understood. Here, rAAV vectors with four-genome lengths were produced using both adherent and suspension HEK293 cells to understand the 5′ITR termination. AAV8 vectors were produced from the human FVIII plasmid for a full-length cargo of 4,707 nucleotides with specific truncations, creating smaller genomes. Conventionally, rAAV is characterized by differentiating empty capsids from full capsids, but for this work, that description is incomplete. The small genomes in this study were characterized by charge detection-mass spectrometry (CD-MS). Using CD-MS, packaged genomes in the range conventionally attributed to partials were resolved and quantified. In addition, alkaline gels and qPCR were used to assess the identity of the packaged genomes. Together, these results showed a propensity for unit-length genomes to be encapsidated. Packaged genomes occurred as replication intermediates emanating from the 5′ITR, indicating that HEK293 cells prefer unit-length genomes as opposed to the 5′ITR termination and heterogeneous DNA packaging observed previously from Sf9 cell systems. As both manufacturing processes are used and continually assessed to produce clinical material, such an understanding will benefit rAAV design for basic research and gene therapy.

The prediction of global horizontal irradiance has become an effective technique to address the intermittence issue of photovoltaic (PV) power generation. This article proposes a novel deep neural network(DNN), named Mixer Dual Path Network (Mixer-DPN), for promising solar forecasting. It shares common features of cloud images and maintains the flexibility to explore new features through dual-path architecture by combining the Mixer layer and Dual Path Network. Therefore, the proposed model can provide more accurate prediction results compared to the classical DNN-based predictors. Moreover, the proposed model shows a faster convergence speed and smaller model size, which makes it suitable for a practical global horizontal irradiance. The merits of the proposed model are verified by testing it with the data from National Renewable Energy Laboratory comparing it with other DNN-based prediction models. Studies have shown that the new model has achieved excellent results in MSE, MAE and other indicators, and the R2 prediction accuracy rate has increased by 14% compared with the baseline model.

Abstract Although more adeno-associated virus AAV-based drugs enter the clinic, vector tissue tropism remains an unresolved challenge that limits its full potential despite that the tissue tropism of naturally occurring AAV serotypes can be altered by genetic engineering capsid vie DNA shuffling, or molecular evolution. To further expand the tropism and thus potential applications of AAV vectors, we utilized an alternative approach that employs chemical modifications to covalently link small molecules to reactive exposed Lysine residues of AAV capsids. We demonstrated that AAV9 capsid modified with N-ethyl Maleimide (NEM) increased its tropism more towards murine bone marrow (osteoblast lineage) while decreased transduction of liver tissue compared to the unmodified capsid. In the bone marrow, AAV9-NEM transduced Cd31, Cd34, and Cd90 expressing cells at a higher percentage than unmodified AAV9. Moreover, AAV9-NEM localized strongly in vivo to cells lining the calcified trabecular bone and transduced primary murine osteoblasts in culture, while WT AAV9 transduced undifferentiated bone marrow stromal cells as well as osteoblasts. Our approach could provide a promising platform for expanding clinical AAV development to treat bone pathologies such as cancer and osteoporosis. Thus, chemical engineering the AAV capsid holds great potential for development of future generations of AAV vectors.

Objective Hook plate fixation is the traditional method for treating distal clavicle fractures. However, in recent years, locked plate applications have emerged as a promising treatment method. This study aimed to compare the short‐ and mid‐term clinical efficacy of anatomical locked plate fixation with coracoclavicular ligament augmentation using anchor nails to that of hook plate fixation in treating distal clavicle fractures. Methods This was a retrospective single‐center cohort study investigating patients with distal clavicle fractures treated between January 2016 and February 2019 in Zhongnan Hospital of Wuhan University. Fifty‐nine eligible patients who underwent either anatomical locked plate fixation with coracoclavicular ligament augmentation using anchor nails (LPF&CLA group; 20 patients) or clavicle hook plate fixation (CHPF group; 39 patients) were included. The visual analog scale (VAS) and Constant–Murley shoulder scores were used to assess shoulder function. In addition, the coracoclavicular distance between the affected and unaffected shoulders (ΔCC distance) was measured to assess the reduction. Patients were followed up at 3 months, 6 months, and 1 year postoperatively. The comparisons between the two groups were made using Student's t‐test, chi‐square test, or Fisher's exact test, if appropriate. Results Preoperative VAS scores were similar in both groups. At 3‐ and 6‐month follow‐up, the VAS score was significantly higher in the CHPF group than in the LPF&CLA group. In contrast, the Constant–Murley shoulder score was significantly lower in the CHPF group than in the LPF&CLA group. When the hook plates were removed, there was no statistical difference in both VAS (0.2 ± 0.4 in LPF&CLA group vs. 0.5 ± 0.5 in CHPF group, p = 0.05) and Constant–Murley shoulder (96.1 ± 3.1 in LPF&CLA group vs. 93.8 ± 5.2 in CHPF group, p = 0.08) scores at the last follow‐up. Postoperatively, the ΔCC distance was 2.37 ± 1.93 mm in the LPF&CLA group and −1.56 ± 1.34 mm in the CHPF group. One year after surgery, ΔCC distance increased to 3.96 ± 1.17 mm in the LPF&CLA group and to −0.89 ± 1.39 mm in the CHPF group. Conclusion For distal clavicle fractures in which the coracoclavicular ligament is disrupted, anatomical locked plate fixation with coracoclavicular ligament augmentation achieved better functional recovery and less pain than hook plate fixation at the 6‐month follow‐up. However, the hook plate provided better reduction throughout the follow‐up period and shoulder pain could be relieved using removal surgery. Therefore, locked plates with coracoclavicular ligament augmentation favors post‐surgery pain relief while harvesting similar functional outcomes to hook plate fixation

Gene therapy is a powerful biological tool that is reshaping therapeutic landscapes for several diseases. Researchers are using both non-viral and viral-based gene therapy methods with success in the lab and the clinic. In the cancer biology field, gene therapies are expanding treatment options and the possibility of favorable outcomes for patients. While cellular immunotherapies and oncolytic virotherapies have paved the way in cancer treatments based on genetic engineering, recombinant adeno-associated virus (rAAV), a viral-based module, is also emerging as a potential cancer therapeutic through its malleability, specificity, and broad application to common as well as rare tumor types, tumor microenvironments, and metastatic disease. A wide range of AAV serotypes, promoters, and transgenes have been successful at reducing tumor growth and burden in preclinical studies, suggesting more groundbreaking advances using rAAVs in cancer are on the horizon.

Adeno-associated virus (AAV) has emerged as a leading platform for gene therapy. With the skyrocketing rate of AAV research and the prevalence of many new engineered capsids being investigated in preclinical and clinical trials, capsid characterization plays a vital role in serotype confirmation and quality control. Further, peptide mapping the capsid proteins might inevitably be a future requirement by regulatory agencies since it is a critical step in good manufacturing practice (GMP) for biotherapeutic characterization. To overcome many challenges that traditional methods like SDS-PAGE and western blots carry, liquid chromatography and mass spectrometry (LC-MS) allows high resolution and sensitivity with great accuracy in characterizing the AAV capsid proteins. Our optimized LC-MS method provides quick sample preparation, a fast and high-throughput 4-min run, and high sensitivity, which allows for very efficient characterization of wild-type and engineered capsids. This study also reports the usage of LC-MS/MS peptide mapping of AAV capsid proteins to determine the most accessible lysine residues targeted by chemical modifications. Our detailed protocols are anticipated to promote the development and discovery of AAV variants with high accuracy and efficiency.

Recombinant adeno-associated virus (rAAV) vectors have been developed for therapeutic treatment of genetic diseases. Current rAAV vectors administered to affected individuals often contain vector DNA-related contaminants. Here we present a thorough molecular analysis of the configuration of non-standard AAV genomes generated during rAAV production using single-molecule sequencing. In addition to the sub-vector genomic-size particles containing incomplete AAV genomes, our results showed that rAAV preparations were contaminated with multiple categories of subgenomic particles with a snapback genome (SBG) configuration or a vector genome with deletions. Through CRISPR and nuclease-based modeling in tissue culture cells, we identified that a potential mechanism leading to formation of non-canonical genome particles occurred through non-homologous end joining of fragmented vector genomes caused by genome lesions or DNA breaks present in the host cells. The results of this study advance our understanding of AAV vectors and provide new clues for improving vector efficiency and safety profiles for use in human gene therapy.

Abstract Background Although ileal faecal diversion is commonly used in clinical settings, complications accompany it. Elucidating the intestinal changes caused by ileal faecal diversion will help resolve postoperative complications and elucidate the pathogenic mechanisms of associated intestinal disorders, such as Crohn's disease (CD). Therefore, our study aimed to provide new insights into the effects of ileal faecal diversion on the intestine and the potential mechanisms. Methods Single‐cell RNA sequencing was performed on proximal functional and paired distal defunctioned intestinal mucosae from three patients with ileal faecal diversion. We also performed in vitro cellular and animal experiments, tissue staining and analysed public datasets to validate our findings. Results We found that the epithelium in the defunctioned intestine tended to be immature, with defective mechanical and mucous barriers. However, the innate immune barrier in the defunctioned intestine was enhanced. Focusing on the changes in goblet cells, we demonstrated that mechanical stimulation promotes the differentiation and maturation of goblet cells through the TRPA1‐ERK pathway, indicating that the absence of mechanical stimulation may be the main cause of defects in the goblet cells of the defunctioned intestine. Furthermore, we found obvious fibrosis with a pro‐fibrotic microenvironment in the defunctioned intestine and identified that monocytes may be important targets for faecal diversion to alleviate CD. Conclusions This study revealed the different transcription landscapes of various cell subsets and the potential underlying mechanisms within the defunctioned intestine, when compared to the functional intestine, based on the background of ileal faecal diversion. These findings provide novel insights for understanding the physiological and pathological roles of the faecal stream in the intestine.

The advent of big data technologies makes a profound impact on various facets of our lives, which also presents an opportunity for Chinese audits. However, the heterogeneity of multi-source audit data, the intricacy of converting Chinese into SQL, and the inefficiency of data processing methods present significant obstacles to the growth of Chinese audits. In this article, we proposed BDMCA, a big data management system designed for Chinese audits. We developed a hybrid management architecture for handling Chinese audit big data, that can alleviate the heterogeneity of multi-mode data. Moreover, we defined an R-HBase spatio-temporal meta-structure for auditing purposes, which exhibits almost linear response time and excellent scalability. Compared to MD-HBase, R-HBase performs 4.5× and 3× better in range query and kNN query, respectively. In addition, we leveraged the slot value filling method to generate templates and build a multi-topic presentation learning model MRo-SQL. MRo-SQL outperforms the state-of-the-art X-SQL parsing model with improvements in logical-form accuracy of up to 5.2%, and execution accuracy of up to 5.9%.

Repurposing clinically approved drugs to construct novel nanomedicines is currently a very attractive therapeutic approach. Selective enrichment of anti-inflammatory drugs and reactive oxygen species (ROS) scavenging at the region of inflammation by stimuli-responsive oral nanomedicine is an effective strategy for the treatment of inflammatory bowel disease (IBD). This study reports a novel nanomedicine, which is based on the excellent drug loading and free radical scavenging ability of mesoporous polydopamine nanoparticles (MPDA NPs). By initiating polyacrylic acid（PAA）polymerization on its surface, a “core-shell” structure nano-carrier with pH response is constructed. Then, under alkaline conditions, using the π-π stacking and hydrophobic interaction between the anti-inflammatory drug sulfasalazine (SAP) and MPDA, the nanomedicines (PAA@MPDA-SAP NPs) loaded efficiently (928 μ g mg−1) of SAP was successfully formed. Our results reveal that PAA@MPDA-SAP NPs can pass through the upper digestive tract smoothly and finally accumulate in the inflamed colon. Through the synergistic effect of anti-inflammation and antioxidation, it can effectively reduce the expression of pro-inflammatory factors and enhance the intestinal mucosal barrier, and finally significantly alleviate the symptoms of colitis in mice. Furthermore, we confirmed that PAA@MPDA-SAP NPs have good biocompatibility and anti-inflammatory repair ability under inflammation induction through human colonic organoids. In summary, this work provides a theoretical basis for the development of nanomedicines for IBD therapy.

Gene therapy offers great promises for a cure of hemophilia A resulting from factor VIII (FVIII) gene deficiency. We have developed and optimized a non-viral ultrasound-mediated gene delivery (UMGD) strategy. UMGD of reporter plasmids targeting mice livers achieved high levels of transgene expression predominantly in hepatocytes. Following UMGD of a plasmid encoding human FVIII driven by a hepatocyte-specific promoter/enhancer (pHP-hF8/N6) into the livers of hemophilia A mice, a partial phenotypic correction was achieved in treated mice. In order to achieve persistent and therapeutic FVIII gene expression, we adopted a plasmid (pHP-hF8-X10) encoding an FVIII variant with significantly increased FVIII secretion. By employing an optimized pulse-train ultrasound condition and immunomodulation, the treated hemophilia A mice achieved 25%–150% of FVIII gene expression on days 1–7 with very mild transient liver damage, as indicated by a small increase of transaminase levels that returned to normal within 3 days. Therapeutic levels of FVIII can be maintained persistently without the generation of inhibitors in mice. These results indicate that UMGD can significantly enhance the efficiency of plasmid DNA transfer into the liver. They also demonstrate the potential of this novel technology to safely and effectively treat hemophilia A.

Prerequisite chains are crucial to acquiring new knowledge efficiently. Many studies have been devoted to automatically identifying the prerequisite relationships between concepts from educational data. Though effective to some extent, these methods have neglected two key factors: most works have failed to utilize domain-related knowledge to enhance pre-trained language models, thus making the textual representation of concepts less effective; they also ignore the fusion of semantic information and structural information formed by existing prerequisites. We propose a two-stage concept prerequisite learning model (TCPL), to integrate the above factors. In the first stage, we designed two continual pre-training tasks for domain-adaptive and task-specific enhancement, to obtain better textual representation. In the second stage, to leverage the complementary effects of the semantic and structural information, we optimized the encoder of the resource–concept graph and the pre-trained language model simultaneously, with hinge loss as an auxiliary training objective. Extensive experiments conducted on three public datasets demonstrated the effectiveness of the proposed approach. Our proposed model improved by 7.9%, 6.7%, 5.6%, and 8.4% on ACC, F1, AP, and AUC on average, compared to the state-of-the-art methods.

The novel gene YBX3 is important for regulating translation and RNA catabolism and encodes a protein with a highly conserved cold-shock domain. However, its pathogenic roles across cancers (e.g., colon cancer) and its regulation remain unclear. We identified the pathogenic roles of YBX3 and its regulatory lncRNA HEIH in various cancers and investigated their effects on tumor progression in colon cancer. Methods including RNA pull-down, MS, and TMA of 93 patients, qPCR of 12 patients with diverse clinicopathologic stages, and western blotting were performed. The pancancer analysis showed that YBX3 expression varies significantly among not only cancer types but also molecular and immune subtypes of the same cancer. Furthermore, its expression in colon cancer is clinically significant, and there is an obvious negative regulatory association between HEIH and YBX3. Among various cancers, especially colon cancer, YBX3 is more related than HEIH expression to the clinical features and prognosis of subgroups. The receiver operating characteristic analysis showed that HEIH and YBX3 have similar predictive capacity in various cancers. The analysis of differentially expressed genes in colon cancer revealed that they have similar hub gene networks, indicating an oncogenic system with a strong overlap. The results also suggest that YBX3 is associated with tumor immune evasion via different mechanisms involving T-cell exclusion in different cancer types and by the tumor infiltration of immune cells. Interestingly, scRNA-seq revealed that HEIH inhibits this phenomenon. Our results also suggest that YBX3 expression is associated with immune or chemotherapeutic outcomes in various cancers, and YBX3 exhibited a higher predictive power than two of seven standardized biomarkers for response outcomes and overall survival of immune checkpoint blockade subcohorts. In colon cancer cell lines, lncRNA-HEIH and YBX3 associate. MS confirmed that YBX3 was pulled down with HEIH, and western blot showed that HEIH knockdown disinhibited YBX3. This study strongly suggests that lncRNA-HEIH/YBX3 is a pancancer immune-oncogenic system and could serve as a biomarker for diagnosis and prognosis and as a therapeutic target, especially in colon cancer.

Mechanosensation is essential for normal gastrointestinal (GI) function, and abnormalities in mechanosensation are associated with GI disorders. There are several mechanosensitive ion channels in the GI tract, namely transient receptor potential (TRP) channels, Piezo channels, two-pore domain potassium (K2p) channels, voltage-gated ion channels, large-conductance Ca2+-activated K+ (BKCa) channels, and the cystic fibrosis transmembrane conductance regulator (CFTR). These channels are located in many mechanosensitive intestinal cell types, namely enterochromaffin (EC) cells, interstitial cells of Cajal (ICCs), smooth muscle cells (SMCs), and intrinsic and extrinsic enteric neurons. In these cells, mechanosensitive ion channels can alter transmembrane ion currents in response to mechanical forces, through a process known as mechanoelectrical coupling. Furthermore, mechanosensitive ion channels are often associated with a variety of GI tract disorders, including irritable bowel syndrome (IBS) and GI tumors. Mechanosensitive ion channels could therefore provide a new perspective for the treatment of GI diseases. This review aims to highlight recent research advances regarding the function of mechanosensitive ion channels in the GI tract. Moreover, it outlines the potential role of mechanosensitive ion channels in related diseases, while describing the current understanding of interactions between the GI tract and mechanosensitive ion channels.

Abstract Predicting the output power of partially‐shaded photovoltaic (PV) systems has a crucial role in the field of photovoltaic system studies and design. A significant number of models, circuit simulation techniques and power prediction formulations are proposed in the literature. However, several factors could limit their application such as computational burden, accuracy and their applicability to different levels of photovoltaic system. An analytical circuit‐based approach is proposed to predict the output power and power losses of partially‐shaded photovoltaic systems. The proposed approach is distinguished by high accuracy, low computational time and applicability to all photovoltaic system levels. Hence, it enables the assessment of partial shading mitigation ability of various photovoltaic system architectures. Furthermore, the proposed approach has no limitations on the size of photovoltaic system, for example, as in circuit‐based simulation analysis. An extensive simulation‐based comparison study for five partially‐shaded photovoltaic systems is conducted to compare the accuracy and computational time of the proposed approach with the detailed, well‐established single diode model. The accuracy of the proposed approach is also validated experimentally on a photovoltaic system of 12 series‐connected photovoltaic modules. Furthermore, a set of comparisons with experimental measurements from the literature is presented.

In the original publication [...]

One important limitation for achieving therapeutic expression of human factor VIII (FVIII) in hemophilia A gene therapy is inefficient secretion of the FVIII protein. Substitution of five amino acids in the A1 domain of human FVIII with the corresponding porcine FVIII residues generated a secretion-enhanced human FVIII variant termed B-domain-deleted (BDD)-FVIII-X5 that resulted in 8-fold higher FVIII activity levels in the supernatant of an in vitro cell-based assay system than seen with unmodified human BDD-FVIII. Analysis of purified recombinant BDD-FVIII-X5 and BDD-FVIII revealed similar specific activities for both proteins, indicating that the effect of the X5 alteration is confined to increased FVIII secretion. Intravenous delivery in FVIII-deficient mice of liver-targeted adeno-associated virus (AAV) vectors designed to express BDD-FVIII-X5 or BDD-FVIII achieved substantially higher plasma FVIII activity levels for BDD-FVIII-X5, even when highly efficient codon-optimized F8 nucleotide sequences were employed. A comprehensive immunogenicity assessment using in vitro stimulation assays and various in vivo preclinical models of hemophilia A demonstrated that the BDD-FVIII-X5 variant does not exhibit an increased immunogenicity risk compared to BDD-FVIII. In conclusion, BDD-FVIII-X5 is an effective FVIII variant molecule that can be further developed for use in gene- and protein-based therapeutics for patients with hemophilia A.

BackgroundCircular RNAs (circRNAs) are a novel type of non-coding RNA, play an important role in the progression of tumors. However, the function and mechanism of circRNAs in regulating immune microenvironment of pancreatic cancer (PC) remain largely unclear.MethodsThe effects of hsa_circ_0046523 expression on proliferation, migration and invasion of PC cells were analyzed by CCK8 and Transwell assays. Flow cytometry was used to detect the proportion of CD4+ T cells, CD8+ T cells and Tregs in peripheral blood mononuclear cells (PBMCs) after co-culture, and the apoptosis, depletion and function of CD8+ T cells. The expression levels of immunoregulatory cytokines were detected by enzyme linked immunosorbent assay (ELISA). The dual-luciferase reporter was performed to determine the interaction between hsa_circ_0046523, miR-148a-3p, and PD-L1. Rescue experiments and PD-L1 blocking experiments were employed to investigate whether hsa_circ_0046523 exerts its biological function by miR-148a-3p/PD-L1 in PC. Furthermore, an immunocompetent murine PC model was established to confirm these findings.ResultsHsa_circ_0046523 expression was remarkably upregulated in PC tissues and cell lines. Moreover, high expression of hsa_circ_0046523 was correlated with advanced pathological stage and poorer prognosis. Hsa_circ_0046523 overexpression promoted the proliferation, migration and invasion of PC cells in vitro. Co-culture experiments confirmed that forced expression of hsa_circ_0046523 could decrease the proportion of CD4+ and CD8+ T cells, as well as increase the proportion of Tregs among peripheral blood mononuclear cells (PBMCs). Meanwhile, hsa_circ_0046523 overexpression promoted the apoptosis and exhaustion of CD8+ T cells, inhibited CD8+ T cell function, increased the secretion of immunosuppressive cytokines IL-10 and TGF-β, and decreased the secretion of immune effector cytokines IFN-γ and IL-2 among PBMCs. Mechanistically, hsa_circ_0046523 exerted its biological function by binding to miR-148a-3p to upregulate PD-L1 expression in PC. Moreover, these immune modulating functions of miR-148a-3p/PD-L1 axis were also confirmed in an immunocompetent murine PC model.ConclusionsOur study suggests that hsa_circ_0046523/miR-148a-3p/PD-L1 regulatory axis mediates PC immunosuppressive microenvironment and these molecules are expected to be new targets for remodeling tumor immune microenvironment of PC.

An ileostomy is a surgery that is commonly performed to protect low pelvic anastomoses or prevent high-risk anastomotic leakages. However, various postoperative complications remain of major concern. After an ileostomy, the distal intestinal segment is left open for an extended period and is in a non-functional state. Consequently, the intestinal mucosa, smooth muscle, and microbiota undergo significant changes that are closely related to postoperative recovery and complications. A systematic description of these changes is necessary to understand the relationship among them and take more effective measures for postoperative intervention.

Physical titers for recombinant adeno-associated viral (rAAV) vectors are measured by quantifying viral genomes. It is generally perceived that AAV virions disassemble and release DNA upon thermal treatment. Here, we present data on enzymatic accessibility of rAAV genomes when AAV virions were subjected to thermal treatment. For rAAV vectors with a normal genome size (≤4.7 kb), thermal treatment at 75°C–99°C allowed only ∼10% of genomes to be detectable by quantitative real-time PCR. In contrast, greater than 70% of AAV genomes can be detected under similar conditions for AAV vectors with an oversized genome (≥5.0 kb). The permeability of virions, as measured by ethidium bromide (EB) staining, was enhanced by thermal stimulation. These results suggest that in rAAV virions with standard-sized genomes, the capsid and DNA are close enough in proximity for heat-induced “crosslinking,” which results in inaccessibility of vector DNA to enzymatic reactions. In contrast, rAAV vectors with oversized genomes release their DNA readily upon thermal treatment. These findings suggested that the spatial arrangement of capsid protein and DNA in AAV virions is genome-size dependent. These results provide a foundation for future improvement of vector assays, design, and applications.

It is well known that canine factor VIII (cFVIII) has a higher specific activity than does human FVIII (hFVIII), and it has been previously demonstrated that cFVIII light chain is able to enhance hFVIII activity. The goal of this study was to first determine which amino acids in cFVIII light chain were responsible for enhancing hFVIII activity, and second to use these amino acids to develop a hFVIII variant with enhanced functional activity. We systemically screened segments of cFVIII light chain by testing an array of human-canine light chain hybrids and found that canine amino acids 1857–2147 were key to this enhancement. Each canine amino acid within this span was screened individually using a negative selection method, which led to the identification of 12 aa (JF12) in the FVIII light chain that could enhance activity. Substitution of the corresponding 12 aa into hFVIII (hFVIIIJF12BDD) elevated the specific activity profile in vitro. Furthermore, hFVIIIJF12BDD expressed an in vivo-displayed increased coagulation activity compared to wild-type, while maintaining normal secretion efficiency. In conclusion, we identified the amino acids in cFVIII that are the key determinants for higher specific activity and may be the basis for future development of therapeutic treatments for hemophilia A.

Fine-grained Named entity recognition (NER) is crucial to natural language processing (NLP) applications like relation extraction and knowledge graph construction. Most existing fine-grained NER systems suffer from inefficiency problem as they use manually annotated training datasets. To address such issue, our NER system could automatically generate datasets from Wikipedia in distant supervision paradigm through mapping hyperlinks in Wikipedia documents to Freebase. In addition, previous NER models can not effectively process fine-grained labels with more than 100 types. So we introduce a `BIO' tagging strategy which can identify the position and type attributes simultaneously. Such tagging scheme transfers NER problem into a sequence-to-sequence (seq2seq) based issue. We propose a seq2seq framework to comprehend the input sentence in a comprehensive way. Specifically, we adopt a Bi-LSTM as the encoder to equally process the past and future information of the input. Then we add a self-attention mechanism to handle the long-term dependency problem in a long sequence. When classifying the entity tags, we choose CRF model as it adds more constraints to avoid position logical problem. Experiments are performed on large-scale datasets for fine-grained NER tasks. Experimental results verify the effectiveness of FSeqC, and it outperforms other state-of-the-art alternatives consistently and significantly.

Current Intellectual Property (IP) copyright protection technologies have low efficiency of authority management, traceability, and scalability. In this work, a blockchain-based IP copyright protection algorithm is proposed to address these issues by establishing a mathematical model of quadratic matrix transformation for IP circuit trading. This algorithm proposes the design of a distributed random embedding mechanism and position mapping function that, when IP trading occurs in blockchain, the traceable mapping function can trace the copyright information in IP trading with the mapping factor. Besides, this work analyzes the credibility, transparency, overhead, and complexity. Experimental results show that the proposed algorithm can resist replaying attacks, yet the copyright information can be rapidly retrieved after suffered from attacks. Still, the proposed algorithm has higher security, stability, and traceability.

Text2SQL can help non-professionals connect with databases by turning natural languages into SQL. Although previous researches about Text2SQL have provided some workable solutions, most of them extract values based on column representation. If there are multiple values in the query and these values belong to different columns, the previous approaches based on column representation cannot accurately extract values. In this work, we propose a new neural network architecture based on the pre-trained BERT, called M-SQL. The column-based value extraction is divided into two modules, value extraction and value-column matching. We evaluate M-SQL on a more complicated TableQA dataset, which comes from an AI competition. We rank first in this competition. Experimental results and competition ranking show that our proposed M-SQL achieves state-of-the-art results on TableQA.

Automatically parsing SQL queries from natural languages can help non-professionals access databases and improve the efficiency of information utilization. It is a long-term research issue and has recently received attention from the relevant communities. Although previous researches have provided some workable solutions, most of them only consider table schemas and natural language questions when parsing SQL queries, and do not use table contents. We observe that table contents can provide more helpful information for some user questions. In this paper, we propose a novel neural network approach, F-SQL, to focus on solving the problem of table content utilization. In particular, we employ the gate mechanism to fuse table schemas and table contents and get the more different representation about table schemas. We test this idea on the WikiSQL and TableQA datasets. Experimental results show that F-SQL achieves new state-of-the-art results on WikiSQL and TableQA.

Oral glutamine (Gln) has been widely used in gastrointestinal (GI) clinical practice, but it is unclear if Ca2+ regulates intestinal Gln transport, although both of them are essential nutrients for mammals. Chambers were used to determine Gln (25 mM)-induced Isc through Na+/Gln co-transporters in the small intestine in the absence or the presence of selective activators or blockers of ion channels and transporters. Luminal but not serosal application of Gln induced marked intestinal Isc, especially in the distal ileum. Lowering luminal Na+ almost abolished the Gln-induced ileal Isc, in which the calcium-sensitive receptor (CaSR) activation were not involved. Ca2+ removal from both luminal and serosal sides of the ileum significantly reduced Gln- Isc. Blocking either luminal Ca2+ entry via the voltage-gated calcium channels (VGCC) or endoplasmic reticulum (ER) release via inositol 1,4,5-triphosphate receptor (IP3R) and ryanodine receptor (RyR) attenuated the Gln-induced ileal Isc, Likewise, blocking serosal Ca2+ entry via the store-operated Ca2+ entry (SOCE), TRPV1/2 channels, and Na+/Ca2+ exchangers (NCX) attenuated the Gln-induced ileal Isc. In contrast, activating TRPV1/2 channels enhanced the Gln-induced ileal Isc. We concluded that Ca2+ signaling is critical for intestinal Gln transport, and multiple plasma membrane Ca2+-permeable channels and transporters play roles in this process. The Ca2+ regulation of ileal Na+/Gln transport expands our understanding of intestinal nutrient uptake and may be significant in GI health and disease.

Early diagnosis and treatment of colorectal cancer (CRC) significantly improves the survival rate and quality of life. Here we screened for differences in glycoproteins associated with tumor-derived exosomes and validated their clinical value to serve as liquid biopsy biomarkers to diagnosed early CRC. Exosomes were extracted from paracancerous tissues, cancer tissues, and plasma. LC-MS/MS proteomic and glycoproteomics analyses were performed using an LTQ-Orbitrap Elite mass spectrometer. The differences in glycoproteins associated with exosomes of paracancerous tissues and cancer tissue were determined, and their levels in plasma exosomes were determined. Statistical analysis was performed to evaluate the diagnostic efficacy of exosome-associated glycoproteins for CRC. We found that the levels of fibrinogen beta chain (FGB) and beta-2-glycoprotein 1 (β2-GP1) in the exosome of CRC tissue were significantly higher compared with those of paracancerous tissues exosome. The areas under the receiver operating characteristic (ROC) curves of plasma exosomal FGB and β2-GP1 as biomarkers for CRC were 0.871 (95% CI = 0.786–0.914) and 0.834 (95% CI = 0.734–0.901), respectively, compared with those of the concentrations of carcinoembryonic antigen concentration [0.723 (95% CI = 0.679–0.853)] and carbohydrate antigen19-9 concentration [0.614 (95% CI = 0.543–0.715)]. Comprehensive proteomics analyses of plasma exosomal biomarkers in CRC identified biomarkers with significant diagnostic efficacy for early CRC, which can be measured using relatively non-invasive techniques.

Since construction involves many stakeholders and their behavioral risk interaction, which brings risks to the project construction, it is necessary to strengthen the research on the risk management of hydropower projects. This study comprehensively considers the characteristics of hydropower project construction and identifies relevant stakeholders to build and improve the stakeholder behavior risk evaluation index system. On this basis, the social network analysis method is used to build an evaluation model of stakeholders’ behavioral risk transmission network, identify core factors and key relationships, analyze the path of behavioral risk transmission, take measures to cut off the transmission of core factors and key relationships, and test the effect of the risk network after control. The results show that: the evaluation model can effectively identify the core behavioral risk factors and key relationships in the construction process. Then, after taking targeted measures on the core behavioral risk factors and key relationships, hydropower projects are less affected by behavioral risk factors, and the risk transmission paths are reduced, which reduces the probability of behavioral risks arising from stakeholders and improves the behavioral governance efficiency of stakeholders. Applying this research model to the risk management of international hydropower projects can provide better guidance to the stakeholders and improve the accuracy and effectiveness of analyzing the behavioral risks of stakeholders in hydropower projects.

Knowledge representation learning is representing entities and relations in a knowledge graph as dense low-dimensional vectors in the continuous space, which explores the features and properties of the graph. Such a technique can facilitate the computation and reasoning on the knowledge graphs, which benefits many downstream tasks. In order to alleviate the problem of insufficient entity representation learning caused by sparse knowledge graphs, some researchers propose knowledge graph embedding models based on instances and concepts, which utilize the latent semantic connections between concepts and instances contained in the knowledge graphs to enhance the knowledge graph embedding. However, they model instances and concepts in the same space or ignore the transitivity of isA relations, leading to inaccurate embeddings of concepts and instances. To address the above shortcomings, we propose a knowledge graph embedding model that differentiates concepts and instances based on spatial transformation—CIST. The model alleviates the gathering issue of similar instances or concepts in the semantic space by modeling them in different embedding spaces, and adds a learnable parameter to adjust the neighboring range for concept embedding to distinguish hierarchical information of different concepts, thus modeling the transitivity of isA relations. The above features of instances and concepts serve as auxiliary information so that thoroughly modeling them could alleviate the insufficient entity representation learning issue. For the experiments, we chose two tasks, i.e., link prediction and triple classification, and two real-life datasets: YAGO26K-906 and DB111K-174. Compared with state of the arts, CIST achieves an optimal performance in most cases. Specifically, CIST outperforms the SOTA model JOIE by 51.1% on Hits@1 in link prediction and 15.2% on F1 score in triple classification.

Human plasma fibronectin is an adhesive protein that plays a crucial role in wound healing. Many studies had indicated that glycans might mediate the expression and functions of fibronectin, yet a comprehensive understanding of its glycosylation is still missing. Here, we performed a comprehensive N- and O-glycosylation mapping of human plasma fibronectin and quantified the occurrence of each glycoform in a site-specific manner. Intact N-glycopeptides were enriched by zwitterionic hydrophilic interaction chromatography, and N-glycosite sites were localized by the 18O-labeling method. O-glycopeptide enrichment and O-glycosite identification were achieved by an enzyme-assisted site-specific extraction method. An RP–LC–MS/MS system functionalized with collision-induced dissociation and stepped normalized collision energy (sNCE)-HCD tandem mass was applied to analyze the glycoforms of fibronectin. A total of 6 N-glycosites and 53 O-glycosites were identified, which were occupied by 38 N-glycoforms and 16 O-glycoforms, respectively. Furthermore, 77.31% of N-glycans were sialylated, and O-glycosylation was dominated by the sialyl-T antigen. These site-specific glycosylation patterns on human fibronectin can facilitate functional analyses of fibronectin and therapeutics development.

Recombinant adeno-associated virus (rAAV) has been developed as a successful vector for both basic research and human gene therapy. However, neutralizing antibodies (NAbs) against AAV capsids can abolish AAV infectivity on target cells, reducing the transduction efficacy. Absence of AAV NAb has become a prerequisite qualification for patients enrolled in gene therapy trials. Nevertheless, accurate assessment of AAV NAb has remained a challenging task. Here we developed a rapid assay based on the observations that AAV NAb inhibits rAAV binding to the host cell surface and NAb titers are negatively related to the amount of AAV genomes binding to the target cells. By quantifying the AAV genome on the target cells in the presence of anti-sera, AAV NAb titers can be accurately determined. The titer determined by this assay correlates well with the classical transduction-based assays. A major advantage of this method is that it can be carried out with a 30-min binding assay without the lengthy wait for a transduction outcome. This assay is independent of transduction performance of AAV serotype in the target cells. Therefore, the AAV cell-binding assay for NAb determination offers an alternative method for in vivo NAb assay. Keywords: AAV, neutralizing antibody assay, gene therapy, vector

Maximum power point tracking (MPPT) is essential in Photovoltaic (PV) systems, which has drawn significant research effort in the past. The operation is to adjust the power interfaces so that the operating characteristics of the consumption and the PV generator match at the ideal level in term of generation. A comprehensive review is essential to help readers understand the latest developments and inform research directions. Unlike the other review papers, this paper focuses on the operational principles of MPPT methods. Therefore, a different review angle is presented in this paper to provide a clear image of the technology of MPPT.

Eco-compensation is an effective means of coordinating ecological protection and economic development, and the assessment of its standards is core content in the study of eco-compensation projects. To improve the operability of eco-compensation standards, taking Nujiang Prefecture as the study area, we combine the equivalent factor method and water footprint method to evaluate the ecosystem-service-value (ESV) spillover and use the market comparison method to calculate the opportunity cost. The final eco-compensation upper and lower limits model is constructed on the basis of the ESV spillover and opportunity cost. The results show the following: (1) the ecological protection of Nujiang Prefecture has been effective, based on the stabilization of its ESV after an initial increase. The main types of ecosystem services provided are regulation and provision services. Gongshan County makes the most significant contribution to the total ESV. (2) The ratio of the ESV self-consumption in Nujiang Prefecture shows a trend of first rising and then falling. This is mainly explained through the reduction in the use of industrial and agricultural water. After deducting self-consumption through the water footprint method, it can be observed that there is ecological spillover in Nujiang Prefecture. (3) The opportunity cost in Nujiang Prefecture increases yearly from 2005 to 2020 owing to ecological protection policies. Combined with the ESV spillover, it is determined that the rational range of the eco-compensation standard is between CNY 6.17 × 102 million and 120.01 × 102 million in 2005, between CNY 10.02 × 102 million and 128.25 × 102 million in 2010, between CNY 30.34 × 102 million and 197.12 × 102 million in 2015, and between CNY 41.97 × 102 million and 227.52 × 102 million in 2020. The current study can offer decision makers a more flexible eco-compensation standard while coordinating the contradiction between regional ecological protection and economic development.

We investigate the eco-environmental effects and the driving factors of transforming the production–living–ecological space (PLES) land use function and offer a scientific foundation for developing regional territorial area and environmental preservation. Eco-environment quality index and ecological contribution ratio are used to analyze the spatial–temporal evolution characteristics and eco-environment effects of land use transformation in the Yangtze River Delta Urban Agglomeration (YRDUA) over the three time periods of 2000, 2010, and 2020, and the geographic detectors are used to analyze the factors that influence the spatial difference of eco-environment quality (EEQ). The findings indicate the following: (1) The land use transformation of YRDUA is primarily shown in the shrinkage of the production land area, the stability of ecological land, and the rapid increase of living land. The area of ecological land, such as water, forest, and pasture, has remained relatively steady from the perspective of secondary land types. In contrast, the area of urban and rural living land has significantly increased. (2) Most land use environment comprises the lower-value zone, accounting for about 50%. The area of the low-value zone has continued to rise owing to the rapid urban and rural living land development, tending to continuous growth. (3) Both the ecological improvement and degradation trends are present simultaneously, although the ecological improvement trend is less prominent than the environmental degradation trend. The primary factor is improving the eco-environment by transforming agricultural production land into forest, water, and ecological pasture land. The degradation of the regional EEQ is mostly due to the occupation of agricultural production land by urban and rural living land. (4) Considering natural elements such as altitude, precipitation, and slope, the extent of land use impacts the EEQ. The combination of several factors has changed the EEQ of the YRDUA. The effect of any two elements is stronger than that of a single factor.

During acute intestinal ischemia reperfusion (IR) injury, the intestinal epithelial barrier (IEB) function is often disrupted. Enteric glial cells (EGCs) play an important role in maintaining the integrity of IEB functions. However, how EGCs regulate IEB function under IR stimulation is unknown. The present study reveals that the adenosine A2A receptor (A2AR) is important for mediating the barrier-modulating roles of EGCs. A2AR knockout (KO) experiments revealed more serious intestinal injury in A2AR KO mice than in WT mice after IR stimulation. Moreover, A2AR expression was significantly increased in WT mice when challenged by IR. To further investigate the role of A2AR in IEB, we established an in vitro EGC-Caco-2 co-culture system. Hypoxia stimulation was used to mimic the process of in vivo IR. Treating EGCs with the CGS21680 A2AR agonist attenuated hypoxia-induced intestinal epithelium damage through up-regulating ZO-1 and occludin expression in cocultured Caco-2 monolayers. Furthermore, we showed that A2AR and metabotropic glutamate receptor 5 (mGluR5) combine to activate the PKCα-dependent pathway in conditions of hypoxia. This study shows, for the first time, that hypoxia induces A2AR-mGluR5 interaction in EGCs to protect IEB function via the PKCα pathway.

Adeno associated viral (AAV) vectors have emerged as a preferred platform for in vivo gene replacement therapy and represent one of the most promising strategies to treat monogenetic disorders such as hemophilia. However, immune responses to gene transfer have hampered human gene therapy in clinical trials. Over the past decade, it has become clear that innate immune recognition provides signals for the induction of antigen-specific responses against vector or transgene product. In particular, TLR9 recognition of the vector’s DNA genome in plasmacytoid dendritic cells (pDCs) has been identified as a key factor. Data from clinical trials and pre-clinical studies implement CpG motifs in the vector genome as drivers of immune responses, especially of CD8+ T cell activation. Here, we demonstrate that cross-priming of AAV capsid-specific CD8+ T cells depends on XCR1+ dendritic cells (which are likely the main cross-presenting cell that cooperates with pDCs to activate CD8+ T cells) and can be minimized by the elimination of CpG motifs in the vector genome. Further, a CpG-depleted vector expressing human coagulation factor IX showed markedly reduced (albeit not entirely eliminated) CD8+ T cell infiltration upon intramuscular gene transfer in hemophilia B mice when compared to conventional CpG+ vector (comprised of native sequences), resulting in better preservation of transduced muscle fibers. Therefore, this deimmunization strategy is helpful in reducing the potential for CD8+ T cell responses to capsid or transgene product. However, CpG depletion had minimal effects on antibody responses against capsid or transgene product, which appear to be largely independent of CpG motifs.