FOXO-regulated DEAF1 controls muscle regeneration through autophagy.

The commonality between various muscle diseases is the loss of muscle mass, function, and regeneration, which severely restricts mobility and impairs the quality of life. With muscle stem cells (MuSCs) playing a key role in facilitating muscle repair, targeting regulators of muscle regeneration has been shown to be a promising therapeutic approach to repair muscles. However, the underlying molecular mechanisms driving muscle regeneration are complex and poorly understood. Here, we identified a new regulator of muscle regeneration, Deaf1 (Deformed epidermal autoregulatory factor-1) - a transcriptional factor downstream of foxo signaling. We showed that Deaf1 is transcriptionally repressed by FOXOs and that DEAF1 targets to Pik3c3 and Atg16l1 promoter regions and suppresses their expression. Deaf1 depletion therefore induces macroautophagy/autophagy, which in turn blocks MuSC survival and differentiation. In contrast, Deaf1 overexpression inactivates autophagy in MuSCs, leading to increased protein aggregation and cell death. The fact that Deaf1 depletion and its overexpression both lead to defects in muscle regeneration highlights the importance of fine tuning DEAF1-regulated autophagy during muscle regeneration. We further showed that Deaf1 expression is altered in aging and cachectic MuSCs. Manipulation of Deaf1 expression can attenuate muscle atrophy and restore muscle regeneration in aged mice or mice with cachectic cancers. Together, our findings unveil an evolutionarily conserved role for DEAF1 in muscle regeneration, providing insights into the development of new therapeutic strategies against muscle atrophy.

Chemsex as lines of flight? Critical Deleuzean perspectives on chemical intimacies.

BACKGROUND: Public health scholars informed by a dominant biomedical paradigm have in their 'rush to risk' emphasised the problematic aspects of chemsex. Meanwhile, critical chemsex scholars have attemped to destigmatise such sexual-chemical practices and foreground how they can be transformative. Taking these two perspectives as points of departure, we make a case for understanding chemsex vis-à-vis Deleuzean lines of flight. METHODS: Semi-structured in-depth interviews were conducted with 33 purposively sampled sexual minority men seeking therapy for chemsex dependency in Singapore. Interview topics included participants' experiences and histories of chemsex, substance use, and their ongoing recovery. Interviews were audio-recorded, transcribed and then analysed according to key themes. RESULTS: We illustrate how chemically inflected sexual encounters can offer deterritorialising flights of fantasy and freedom from a heteronormative social structure that pathologises gay sex. At the same time, we argue that these flight lines are not static, neither do they extend indefinitely in space-time. Rather, we show how flights of freedom can evolve into lines of fright (or non-flight) when chemsex practitioners are met with critical thresholds that reveal the less-than-desirable aspects of being intoxicated. Consequently, they may eventually consider the reterritorialisation of their lives (i.e. abstinence and recovery) as a more constructive option. Regardless of their decisions to ride on chemically-induced flight lines or to take a step back from such deterritorialising pathways, they have troubled stereotypical perspectives of drug users as passive automatons. CONCLUSIONS: This paper enriches the chemsex scholarship by presenting a Deleuzean conceptualisation of chemical-sexual intimacies without romantacising and/or overstating the 'escape'/'freedom' that chemsex can facilitate. Future research in this arena could explore the complicated intimate relationships that users may have with their drug(s) of choice, and their varied lines of (non-)flight over a longitudinal study.

Biogeography and impact of nitrous oxide reducers in rivers across a broad environmental gradient on emission rates.

Microbial communities that reduce nitrous oxide (N2O) are divided into two clades, nosZI and nosZII. These clades significantly differ in their ecological niches and their implications for N2O emissions in terrestrial environments. However, our understanding of N2O reducers in aquatic systems is currently limited. This study investigated the relative abundance and diversity of nosZI- and nosZII-type N2O reducers in rivers and their impact on N2O emissions. Our findings revealed that stream sediments possess a high capacity for N2O reduction, surpassing N2O production under high N2O/NO3- ratio conditions. This study, along with others in freshwater systems, demonstrated that nosZI marginally dominates more often in rivers. While microbes containing either nosZI and nosZII were crucial in reducing N2O emissions, the net contribution of nosZII-containing microbes was more significant. This can be attributed to the nir gene co-occurring more frequently with the nosZI gene than with the nosZII gene. The diversity within each clade also played a role, with nosZII species being more likely to function as N2O sinks in streams with higher N2O concentrations. Overall, our findings provide a foundation for a better understanding of the biogeography of stream N2O reducers and their effects on N2O emissions.

Cardiolipin oxidized by ROS from complex II acts as a target of gasdermin D to drive mitochondrial pore and heart dysfunction in endotoxemia.

Cardiac dysfunction, an early complication of endotoxemia, is the major cause of death in intensive care units. No specific therapy is available at present for this cardiac dysfunction. Here, we show that the N-terminal gasdermin D (GSDMD-N) initiates mitochondrial apoptotic pore and cardiac dysfunction by directly interacting with cardiolipin oxidized by complex II-generated reactive oxygen species (ROS) during endotoxemia. Caspase-4/11 initiates GSDMD-N pores that are subsequently amplified by the upregulation and activation of NLRP3 inflammation through further generation of ROS. GSDMD-N pores form prior to BAX and VDAC1 apoptotic pores and further incorporate into BAX and VDAC1 oligomers within mitochondria membranes to exacerbate the apoptotic process. Our findings identify oxidized cardiolipin as the definitive target of GSDMD-N in mitochondria of cardiomyocytes during endotoxin-induced myocardial dysfunction (EIMD), and modulation of cardiolipin oxidation could be a therapeutic target early in the disease process to prevent EIMD.

Effects of carbon to nitrogen ratio on nitrogen removal in a single-stage microaerobic system: A model-based evaluation.

Single-stage microaerobic systems have been proven to be effective for concurrent removal of ammonium and organic carbon from sewage. While mechanistic models derived from activated sludge models (ASMs) have simulated nutrients removal under microaerobic conditions, classic ASMs exhibit limitations in capturing the intricate effects of carbon to nitrogen (C/N) ratio on nitrogen removal performance. To address this issue, a mechanistic model modified from the classic ASMs was proposed to capture the combined inhibitory effects of carbon and ammonium on microaerobic systems. This modified model was established based on experimental data from a single-stage microaerobic reactor encompassing simultaneous nitrification-denitrification and anammox processes. The inhibition coefficient of C/N ratio was integrated into the process rate equations, and its effectiveness was validated through model performance evaluation. Compared to the classic models, the modified one achieved superior predictions for nitrite and nitrate nitrogen concentrations. Simulations revealed that under optimized conditions with a C/N of 4.57 and a dissolved oxygen (DO) of 0.41 mg/L, the system could achieve up to 95.5% of total nitrogen (TN) removal efficiency. Based on the simulation of substrate uptake/production rate, increasing the nitrogen loading rate (NLR) rather than organic loading rate (OLR) was crucial for efficient nitrogen removal. The proposed modified model served as a valuable tool for designing and optimizing similar biological wastewater treatment systems.

Comparative study of different extendable intramedullary rods combined with surgery in the treatment of congenital pseudarthrosis of the tibia.

BACKGROUND: When using traditional extensible intramedullary rods to treat congenital pseudarthrosis of the tibia (CPT), there were cases of re-fracture and internal fixation fracture. Therefore, the authors propose a research hypothesis that a thicker distal extensible intramedullary rod can better protect the tibia and reduce the incidence of refracture PURPOSE: To investigate the clinical efficacy of new and traditional extensible intramedullary rods in the treatment of CPT in children METHODS: From January 2017 to December 2021, the clinical data of 49 children with CPT who were treated with traditional extensible intramedullary rod combined surgery (group A) and new extensible intramedullary rod combined surgery (group B) in our hospital were collected. Inclusive criteria: ① Crawford type IV CPT children; ② The operation was performed by the same team. EXCLUSION CRITERIA: patients with multiple tibial angulation. During follow-up, the initial healing, proximal tibial valgus, tibial length, ankle valgus, refracture and intramedullary rod displacement of CPT children in the two groups were evaluated RESULTS: It was a retrospective investigation. In group A, 26 cases met the inclusion criteria, 24 cases achieved primary healing, with an primary healing rate of 92%, including 1 case of nonunion due to osteomyelitis complications after surgery, and 1 case of delayed healing, with an average healing time of 4.7 ± 0.8 months. 17 cases (68%) had unequal tibia length, with an average difference of 1.6 ± 0.8 cm. Ankle valgus occurred in 10 cases (40%) with an average of 14.4°±4.8°; Proximal tibial valgus occurred in 6 cases (24%) with an average of 7 °± 1.8 °. 20 cases (80%) had tip of the rod migration.10 cases (40%) had re-fracture; The average follow-up time was 2.4 ± 0.4 years. In group B, 22 patients achieved primary healing, and the primary healing rate was 95%, including 1 case with delayed healing. The average healing time was 4.7 ± 1.7months. 14 cases (61%) had unequal tibia length, with an average difference of 1 ± 0.5 cm. Ankle valgus occurred in 4 cases (17%) with an average of 12.3 °±4.9°; The proximal tibia valgus occurred in 9 cases (39%), with an average of 7.7 °±2.5 °. 14 cases (61%) had new type of intramedullary rod displacement. 3 cases (13%) had re-fracture; The average follow-up time was 2.3 ± 0.6years CONCLUSION: Compared with the traditional extended intramedullary rod combined operation, the new type of extended intramedullary rod combined operation has a lower incidence of re-fracture after CPT, but it still needs to be verified by large sample and multi-center research.

Effect of hydrothermal-acid pretreatment on methane yield and microbial community in anaerobic digestion of rice straw.

Hydrothermal pretreatment has been proposed to enhance straw methane yield during anaerobic digestion recently. However, the combined effect of hydrothermal and organic acid pretreatment (HTOAP) needs further investigation. This study identified optimal pretreatment at 120 °C with 3 % acetic acid for 24 h by orthogonal design method. The HTOAP increased the reducing sugar content by destroying the lignocellulosic structure. A 79 % increment of methane production after HTOAP was observed compared to the untreated group. Microbial analysis showed that HTOAP enriched the relative abundance of lignocellulose-degraders, such as W5053, Thermanaerovibrio, Caldicoprobacter, as well as the syntrophic acetate oxidizing bacteria Syntrophaceticus. Moreover, Methanobacterium conducted hydrogenotrophic methanogenesis dominantly. Furthermore, the potential function analysis showed that HTOAP stimulated the expression of key enzymes in the hydrogenotrophic pathway, including carbon-monoxide dehydrogenase (EC 1.2.7.4) and coenzyme F420 hydrogenase (EC 1.12.98.1). This investigation illustrated the potential of HTOAP of rice straw to facilitate methane production.

METTL3 promotes the osteogenic differentiation of periosteum-derived MSCs via regulation of the HOXD8/ITGA5 axis in congenital pseudarthrosis.

Background: Congenital pseudarthrosis of the tibia (CPT) is a dominant health challenge in pediatric orthopedics. The essential process in the development of CPT is the limited capacity of mesenchymal stem cells (MSCs) derived from CPT to undergo osteogenic differentiation. Our research aimed to elucidate the role and mechanism of methyltransferase-like 3 (METTL3) in the osteogenic differentiation process of CPT MSCs. Methods: The osteogenic differentiation medium was used to culture MSCs, and the detection of osteogenic differentiation was performed using Alizarin Red S and alkaline phosphatase (ALP) assays. Gene or protein expression was assessed by quantitative real-time polymerase chain reaction (qRT-PCR), Western blot, or immunofluorescence (IF) staining. The m6A modification of Homeobox D8 (HOXD8) was verified by methylated RNA immunoprecipitation (MeRIP) assay. Interactions between METTL3 and HOXD8 or HOXD8 and integrin alpha 5 (ITGA5) promoter were validated by the luciferase reporter gene, RIP, and chromatin immunoprecipitation (ChIP) assays. Results: METTL3 overexpression enhanced CPT MSCs' osteogenic differentiation. METTL3 stabilized the HOXD8 in an m6A-dependent manner. Moreover, the overexpressed ITGA5 up-regulated the CPT MSCs' osteogenic differentiation. Further, HOXD8 could transcriptionally activate ITGA5. METTL3 increased the transcription of ITGA5 via HOXD8 to enhance the osteogenic differentiation of CPT MSCs. Conclusion: METTL3 promoted osteogenic differentiation via modulating the HOXD8/ITGA5 axis in CPT MSCs.

Global trends and hotspots in research on acupuncture for neurodegenerative diseases over the last decade: a bibliometric and visualization analysis.

Objectives: This study aimed to explore the current status and trends of acupuncture for neurodegenerative diseases (NDs) in the last decade and provide new insights for researchers in future studies. Methods: The publications concerning acupuncture treatment for NDs published between 2014 and 2023 were extracted from the Web of Science Core Collection. We used CiteSpace and VOSviewer to analyze data on numbers of annual publications, countries, institutions, cited journals, cited authors, cited references, keywords, and citation bursts about acupuncture for NDs. Results: A total of 635 publications were obtained from 2014 to 2023. We identified the most prolific journals, countries, institutions, authors, patterns of authorship, and the main direction of future research in the field of acupuncture for NDs in the last decade. The country, institution, and journal with the most publications are China (389 articles), Beijing University of Chinese Medicine (56 articles), and Evidence Based Complementary and Alternative Medicine (42 articles), respectively. The high-frequency keywords focused on "Alzheimer's disease," "Parkinson's disease," "acupuncture," "dementia," and "electroacupuncture." The top five keywords in terms of centrality were "cerebral ischemia," "acupuncture stimulation," "fMRI," "apoptosis," and "deep brain stimulation." Conclusion: The results from this bibliometric study provide insight into the research trends in acupuncture therapy for NDs, and the current status and trends of the past decade, which may help researchers confirm the current status, hotspots, and frontier trends in this field.

ggbulksurv: An R package for easy Drosophila and C. elegans survival analysis.

Lifespan studies on fast-aging model organisms like C.elegans and D.melanogaster are conducted with multiple organisms per vial. Lifespan data results in a "one row, multiple individuals" format, which is incompatible with R packages that require a "one row, one individual" format. We present ggbulksurv , an R package for user-friendly survival analysis and highlight three key features. (1) pivot_prism converts data for PRISM, allowing biologists to plot survival curves without manually expanding each observation. (2) run_bulksurv() takes in a "one row, multiple individuals" table and plots a customizable survival curve. (3) Advanced users who require custom survival objects can specify a custom formula, facilitating complex survival analysis. We provide a time saving solution for lifespan data analysis.

Resveratrol activates autophagy and protects from UVA-induced photoaging in human skin fibroblasts and the skin of male mice by regulating the AMPK pathway.

Skin photoaging is mostly caused by ultraviolet A (UVA), although active medications to effectively counteract UVA-induced photoaging have not yet been created. Resveratrol, a naturally occurring polyphenol found in the skin of grapes, has been shown to have various biological functions such as anti-inflammatory and antioxidant characteristics. However, the role of resveratrol in UVA-induced photoaging has not been clarified. We investigated the mechanism of action of resveratrol by UVA irradiation of human skin fibroblasts (HSF) and innovatively modified a mouse model of photoaging. The results demonstrated that resveratrol promoted AMP-activated protein kinase (AMPK) phosphorylation to activate autophagy, reduce reactive oxygen species (ROS) production, inhibit apoptosis, and restore normal cell cycle to alleviate UVA-induced photoaging. In addition, subcutaneous injection of resveratrol not only improved the symptoms of roughness, erythema, and increased wrinkles in the skin of UVA photodamaged mice, but also alleviated epidermal hyperkeratosis and hyperpigmentation, reduced inflammatory responses, and inhibited collagen fiber degradation. In conclusion, our studies proved that resveratrol can treat UVA-induced photoaging and elucidated the possible molecular mechanisms involved, providing a new therapeutic strategy for future anti-aging.

Application and effect of tension-reducing suture in surgical treatment of hypertrophic scar.

PURPOSE: To investigate the application and effectiveness of tension-reducing suture in the repair of hypertrophic scars. METHODS: A retrospective analysis of clinical data was conducted on 82 patients with hypertrophic scars treated at the Department of Burns and Plastic Surgery of Nanjing Drum Tower Hospital from September 2021 to December 2022. Patients were operated with combination of heart-shaped tension-reducing suturing technique and looped, broad, and deep buried (LBD) suturing technique or conventional suture method. Outcomes of surgical treatment were assessed before and 6 months after surgery using the Patient and Observer Scar Assessment Scale (POSAS) and the Vancouver Scar Scale (VSS). RESULTS: Improvements were achieved on scar quality compared to that preoperatively, with a reduction in scar width (1.7 ± 0.6 cm vs. 0.7 ± 0.2 cm, P < 0.001). Assessment using the POSAS and VSS scales showed significant improvements in each single parameter and total score compared to preoperative values (P < 0.05). The Combination method group achieved better score in total score of VSS scale, in color, stiffness, thickness and overall opinion of PSAS scale, and in vascularity, thickness, pliability and overall opinion of OSAS scale. CONCLUSION: The amalgamation of the heart-shaped tension-reducing suturing technique and the LBD suturing technique has shown promising outcomes, garnering notably high levels of patient satisfaction in the context of hypertrophic scar repair. Patients have exhibited favorable postoperative recoveries, underscoring the clinical merit and the prospective broader applicability of this approach in the realm of hypertrophic scar management.

Oncolytic virotherapy stimulates anti­tumor immune response and demonstrates activity in advanced sarcoma: Report of two cases.

Sarcoma is derived from mesenchymal neoplasms and has numerous subtypes, accounting for 1% of all adult malignancies and 15% of childhood malignancies. The prognosis of metastatic or recurrent sarcoma remains poor. The current study presents two cases of sarcoma enrolled in a phase I dose escalation trial for solid tumor, who had previously failed all standard therapies. These patients were treated with VG161, an immune-stimulating herpes simplex virus type 1 oncolytic virus with payloads of IL-12, IL-15 and IL-15 receptor α unit, and a programmed cell death 1 (PD-1)/PD-1 ligand 1 blocking peptide. Both cases demonstrated stable disease as the best response, accompanied by a noteworthy prolongation of progression-free survival (11.8 months for chondrosarcoma and 11.9 months for soft tissue sarcoma, respectively) at a dose of 2.5×108 PFU/cycle. In addition, the treatment led to the activation of anti-cancer immunity, as evident from cytokine, lymphocyte subset and related pathway analyses of peripheral blood and/or tumor biopsy samples. These promising results suggest that VG161 monotherapy holds promise as an effective treatment for sarcoma and warrants further investigation through clinical trials. The two reported patients were part of a phase I clinical trial conducted and registered on the Australian New Zealand Clinical Trials Registry in Australia (registration no. ACTRN12620000244909; registration date, 26 February, 2020).

Land-use simulation for synergistic pollution and carbon reduction: Scenario analysis and policy implications.

Simulations of sustainable land use and management are required to achieve targets to reduce pollution and carbon emissions. Limited research has been conducted on synergistic pollution and carbon reduction (SPCR) in land-use simulations. This study proposed a framework for land-use simulation focused on SPCR. The non-dominated sorting genetic algorithm (NSGA-Ⅱ) and the entropy weight-based technique for order of preference by similarity to an ideal solution (TOPSIS) were used to optimize the land-use structure according to minimum net carbon, nitrogen, and phosphorus emissions. The cellular automata (CA) Markov model was then utilized to simulate the land-use spatial pattern according to the optimal conditions. The proposed framework was applied to the Dongjiang River Basin, South China, and three other scenarios (natural development (ND), carbon minimization (CM), and pollution minimization (PM)) were designed to validate the effectiveness of pollution and carbon emissions reduction under the SPCR scenario. The land-use structure and the pollution and carbon emissions in the scenarios were compared. The results showed the following. (1) The proportions of cultivated land, woodland, grassland, water, and construction land In the SPCR scenario accounted for 14%, 72%, 4%, 3%, and 7% of the total area, respectively. The carbon, nitrogen, and phosphorus emissions were 42.4%, 6.6%, and 7.8% lower, respectively, in the SPCR scenario than in the ND scenario, demonstrating the advantages of simultaneous pollution and carbon reduction. (2) The kappa coefficient of the CA-Markov model was 0.8729, indicating high simulation accuracy. (3) The simulated land-use spatial patterns exhibited low spatial heterogeneity under the CM, PM, and SPCR scenarios. However, there were significant disparities between the ND and SPCR scenarios. The cultivated and construction land areas were significantly smaller in the SPCR scenario than in the ND scenario. In contrast, the woodland and grassland areas were larger, with most differences in the central and southwestern regions of the Dongjiang River Basin. The results of the current study can be used to formulate effective land use policies and strategies in the Dongjiang Basin and similar areas to achieve the Coupling coordination between pollution reduction and carbon reduction. Policy recommendations include increasing the proportion of woodland and grassland, implementing reasonable constraints on expanding cultivated and construction lands, and establishing farmland red lines to promote synergistic pollution and carbon reduction.

Systematic evaluation of machine learning-enhanced trifocal IOL power selection for axial myopia cataract patients.

PURPOSE: This study aimed to evaluate and optimize intraocular lens (IOL) power selection for cataract patients with high axial myopia receiving trifocal IOLs. DESIGN: A multi-center, retrospective observational case series was conducted. Patients having an axial length ≥26 mm and undergoing cataract surgery with trifocal IOL implanted were studied. METHODS: Preoperative biometric and postoperative outcome data from 139 eyes were collected to train and test various machine learning (ML) models (support vector machine, linear regression, and stacking regressor) using five-fold cross-validation. The models' performance was further validated externally using data from 48 eyes enrolled from other hospitals. Performance of seven IOL calculation formulas (BUII, Kane, EVO, K6, DGS, Holladay I, and SRK/T) were examined with and without ML models. RESULTS: The results of cross-validation revealed improvements across all IOL calculation formulas, especially for K6 and Holladay I. The model increased the percentage of eyes with a prediction error (PE) within ±0.50 D from 71.94% to 79.14% for K6, and from 35.25% to 51.80% for Holladay I. In external validation involving 48 patients from other centers, six out of seven formulas demonstrated a reduction in the mean absolute error (MAE). K6's PE within ±0.50 D improved from 62.50% to 77.08%, and Holladay I from 16.67% to 58.33%. CONCLUSIONS: In this study, we conducted a comprehensive evaluation of seven IOL power calculation formulas in high axial myopia cases and explored the effectiveness of the Stacking Regressor model in augmenting their accuracy. Of these formulas, K6 and Holladay I exhibited the most significant improvements, suggesting that integrating ML may have varying levels of effectiveness across different formulas but holds substantial promise in improving the predictability of IOL power calculations in patients with long eyes.

Successful reimplantation of extruded bone segment in lower limb open fractures: case report and literature review.

Objective: The aim of this study is to summarize and demonstrate the different sterilization methods and surgical techniques for open fractures with impacted bone segments in the lower limbs. Methods: A retrospective analysis was conducted on the clinical characteristics, treatment methods, and outcomes of a case involving a 10.5 cm extruded segment of the femur in a 9-year-old male with a right femoral comminuted fracture treated at our center. Additionally, a retrospective review and summary were conducted on all reported cases of open fractures with impacted bone segments in the lower limbs. Results: Our center treated a 9-year and 11-month-old male child who presented with a Gustilo type IIIB open fracture of the femur along with a large segment of the femur being ejected as a result of a car accident. The child was resuscitated to correct hypovolemic shock, underwent emergency wound debridement, and had Ilizarov external fixation of the femur. The ejected femur segment was sterilized using ethylene oxide and re-implanted four days after the injury. A literature review showed that out of the cases of open fractures with impacted bone segments in the lower limbs, there were 14 cases involving the femur and 5 cases involving the tibia. Among them, sterilization was performed using povidone-iodine in 6 cases, high-pressure steam sterilization in 3 cases, and other methods including gamma-ray irradiation and soaking in antibacterial solution were used in the remaining cases. In terms of surgical methods, 7 cases were fixed with locking plates, 3 cases were fixed with external fixation devices, 1 case was immobilized in a cast, 1 case was fixed with an intramedullary rod, and 4 cases involved a combination of external fixation and internal fixation. The average time for re-implantation was 7.6 days after the injury. There were no serious complications such as infection or non-union observed in any of the cases during follow-up. Conclusion: Ethylene oxide can be considered a reliable choice for the reimplantation of displaced bone segments in open fractures after sterilization.

The Wnt Co-Receptor PTK7/Otk and Its Homolog Otk-2 in Neurogenesis and Patterning.

Wnt signaling is a highly conserved metazoan pathway that plays a crucial role in cell fate determination and morphogenesis during development. Wnt ligands can induce disparate cellular responses. The exact mechanism behind these different outcomes is not fully understood but may be due to interactions with different receptors on the cell membrane. PTK7/Otk is a transmembrane receptor that is implicated in various developmental and physiological processes including cell polarity, cell migration, and invasion. Here, we examine two roles of Otk-1 and Otk-2 in patterning and neurogenesis. We find that Otk-1 is a positive regulator of signaling and Otk-2 functions as its inhibitor. We propose that PTK7/Otk functions in signaling, cell migration, and polarity contributing to the diversity of cellular responses seen in Wnt-mediated processes.

Bioactive triterpenoid compounds of Poria cocos (Schw.) Wolf in the treatment of diabetic ulcers via regulating the PI3K-AKT signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic ulcers represent a chronic condition characterized by prolonged hyperglycemia and delayed wound healing, accompanied by endocrine disorders, inflammatory responses, and microvascular damage in the epidermal tissue, demanding effective clinical treatment approaches. For thousands of years, ancient Chinese ethnopharmacological studies have documented the use of Poria cocos (Schw.) Wolf in treating diabetic ulcers. Recent research has substantiated the diverse pharmacological effects of Poria cocos (Schw.) Wolf, including its potential to alleviate hyperglycemia and exhibit anti-inflammatory, antioxidant, and immune regulatory properties, which could effectively mitigate diabetic ulcer symptoms. Furthermore, being a natural medicine, Poria cocos (Schw.) Wolf has demonstrated promising therapeutic effects and safety in the management of diabetic ulcers, holding significant clinical value. Despite its potential clinical efficacy and applications in diabetic ulcer treatment, the primary active components and underlying pharmacological mechanisms of Poria cocos (Schw.) Wolf remains unclear. Further investigations are imperative to establish a solid foundation for drug development in this domain. AIM OF THE STUDY AND MATERIALS AND METHODS: In this study, we aimed to identify the active compounds and potential targets of Poria cocos (Schw.) Wolf using UHPLC-Q-TOF-MS and TCMSP databases. Additionally, we attempt to identify targets related to diabetic ulcers. Following enrichment analysis, a network of protein-protein interactions was constructed to identify hub genes based on the common elements between the two datasets. To gain insights into the binding activities of the hub genes and active ingredients, molecular docking analysis was employed. Furthermore, to further validate the therapeutic effect of Poria cocos (Schw.) Wolf, we exerted in vitro experiments using human umbilical vein vascular endothelial cells and human myeloid leukemia monocytes (THP-1). The active ingredient of Poria cocos (Schw.) Wolf was applied in these experiments. Our investigations included various assays, such as CCK-8, scratch test, immunofluorescence, western blotting, RT-PCR, and flow cytometry, to explore the potential of Poria cocos (Schw.) Wolf triterpenoid extract (PTE) in treating diabetic ulcers. RESULTS: The findings here highlighted PTE as the primary active ingredient in Poria cocos (Schw.) Wolf. Utilizing network pharmacology, we identified 74 potential targets associated with diabetic ulcer treatment for Poria cocos (Schw.) Wolf, with five hub genes (JUN, MAPK1, STAT3, AKT1, and CTNNB1). Enrichment analysis revealed the involvement of multiple pathways in the therapeutic process, with the PI3K-AKT signaling pathway showing significant enrichment. Through molecular docking, we discovered that relevant targets within this pathway exhibited strong binding with the active components of Poria cocos (Schw.) Wolf. In vitro experiments unveiled that PTE (10 mg/L) facilitated the migration of human umbilical vein vascular endothelial cells (P < 0.05). PTE also increased the expression of CD31 and VEGF mRNA (P < 0.05) while activating the expressions of p-PI3K and p-AKT (P < 0.05). Moreover, PTE demonstrated its potential by reducing the expression of IL-1ß, IL-6, TNF-α, and NF-κB mRNA in THP-1 (P < 0.05) and fostering M2 macrophage polarization. These results signify the potential therapeutic effects of PTE in treating diabetic ulcers, with its beneficial actions mediated through the PI3K-AKT signaling pathway. CONCLUSIONS: PTE is the main active ingredient in Poria cocos (Schw.) Wolf that exerts therapeutic effects. Through PI3K-AKT signaling pathway activation and inflammatory response reduction, PTE promotes angiogenesis, thereby healing diabetic ulcers.

Eutrophication-driven infochemical dimethylsulfide accelerates carbon transfer in freshwater food chain.

Dimethylsulfide (DMS) is a major organic sulfide in aquatic ecosystems and an infochemical that is considered as a key predictor of changes in energy and material fluxes and stocks. It is largely unknown how DMS changes and affects the food webs and material cycles in eutrophicated freshwater. In this study, field monitoring and literature surveys were conducted to analyze the effects of eutrophication on DMS concentrations. Daphnia-zebrafish microcosms were then used to investigate the effects of DMS concentrations on carbon transfer. The results demonstrated that the concentration of DMS was increased by eutrophication related indicators (chlorophyll and phosphorus). Eutrophication driven DMS altered carbon transfer in the freshwater food chain. Low concentrations (0.1-1 nM) of DMS promoted the predation of daphnia by zebrafish compared to the 0.01 nM DMS, which further stimulated the total carbon transfer from daphnia to zebrafish and altered the dissolved organic carbon (DOC) distribution in water. High concentrations (10-100 nM) of DMS did not alter zebrafish predation on daphnia and carbon transfer. DOC excreted by zebrafish altered carbon emission potential, and DMS in water showed a unimodal relationship with the carbon emission potential, peaking at 0.40 nM DMS. Keeping the DMS in water at 1.82 nM may maintain a lower carbon emission potential. These results improved the understanding of the effects of eutrophication on DMS, demonstrated the ecological role of DMS on freshwater fish and the carbon cycle, estimated the effects of DMS on the carbon emission potential of fish, and offered new insights into the management of eutrophication.