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Abd Wahid Rizaldi Akili,1 Nisrina Azizah Thurfah,1 Ari Hardianto,1 Jalifah Latip,2 Tati Herlina1 1Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Padjadjaran, Sumedang, West Java, 45363, Indonesia; 2Department of Chemical Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia (UKM), Bangi, Selangor, 46300, MalaysiaCorrespondence: Tati Herlina, Email tati.herlina@unpad.ac.idPurpose: This study aimed to evaluate the potency of 471 flavonoids from the genus Erythrina as potential acetylcholinesterase (AChE) inhibitors and free radical scavengers through computational studies to develop Alzheimer’s disease (AD) therapies from natural products.Methods: A total of 471 flavonoids from the genus Erythrina were subjected to molecular docking against AChE, followed by toxicity screening. The potential AChE inhibitors with the least toxic profile were subjected to further investigation through molecular dynamics (MD) simulations, density functional theory (DFT) study, and in silico pharmacokinetic predictions.Results: A combination of molecular docking and in silico toxicity screening led to the identification of 2(S)‒5,7‒dihydroxy‒5′‒methoxy‒[2′′,2′′‒(3′′‒hydroxy)‒dimethylpyrano]‒(5′′,6′′:3′,4′) flavanone (89) and Abyssinoflavanone IV (83) as potential AChE inhibitors. These compounds had stable binding to AchE and exhibited lower Root Mean Square Deviation (RMSD) values compared to the apo state of AChE. In addition, Molecular Mechanics Generalized Born Surface Area (MMGBSA) analysis revealed that the binding energies of 89 and 83 were significantly lower compared to acetylcholine, the natural substrate of AChE. Based on DFT study, these compounds exhibited a higher energy in the highest occupied molecular orbital (EHOMO) and lower electron affinity (EA) than Quercetin. This indicated that 89 and 83 could be potential radical scavengers through their electron-donating activity.Conclusion: Although this study primarily relied on computational methods, the results showed the dual functionality of compounds 89 and 83 as both potential AChE inhibitors and free radical scavengers. Further investigation in wet laboratory experiments is required to validate their therapeutic potential for AD.Keywords: flavonoids, erythrina, acetylcholinesterase, free radical scavenger, in silico

Abstract Drug-induced liver injury (DILI) refers to drug-mediated damage to the structure and function of the liver, ranging from mild elevation of liver enzymes to severe hepatic insufficiency, and in some cases, progressing to liver failure. The mechanisms and clinical symptoms of DILI are diverse due to the varying combination of drugs, making clinical treatment and prevention complex. DILI has significant public health implications and is the primary reason for post-marketing drug withdrawals. The search for reliable preclinical models and validated biomarkers to predict and investigate DILI can contribute to a more comprehensive understanding of adverse effects and drug safety. In this review, we examine the progress of research on DILI, enumerate in vitro models with potential benefits, and highlight cellular molecular perturbations that may serve as biomarkers. Additionally, we discuss omics approaches frequently used to gather comprehensive datasets on molecular events in response to drug exposure. Finally, three commonly used gene modulation techniques are described, highlighting their application in identifying causal relationships in DILI. Altogether, this review provides a thorough overview of ongoing work and approaches in the field of DILI.

Abstract Ribosomal proteins (RPs) are essential components of ribosomes, playing a role not only in ribosome biosynthesis, but also in various extra-ribosomal functions, some of which are implicated in the development of different types of tumors. As universally acknowledged, hepatocellular carcinoma (HCC) has been garnering global attention due to its complex pathogenesis and challenging treatments. In this review, we analyze the biological characteristics of RPs and emphasize their essential roles in HCC. In addition to regulating related signaling pathways such as the p53 pathway, RPs also act in proliferation and metastasis by influencing cell cycle, apoptosis, angiogenesis, and epithelial-to-mesenchymal transition in HCC. RPs are expected to unfold new possibilities for precise diagnosis and individualized treatment of HCC.

Background: In acute coronary syndrome (ACS), antiplatelet therapy is crucial for inhibiting platelet aggregation. Dual antiplatelet therapy (DAPT) commonly employs aspirin along with clopidogrel, prasugrel, or ticagrelor. This is well known that aspirin acts as a cyclooxygenase (COX)-1 inhibitor, while clopidogrel, prasugrel, and ticagrelor act as P2Y12 inhibitors. Despite DAPT's proven efficacy in more effectively reducing cardiovascular events in ACS patients, this is associated with an increased risk of bleeding compared to mono antiplatelet therapy (MAPT). To minimize the cost and side effect that might arise from the use of DAPT, this is necessary to assess the potential of MAPT using a P2Y12 inhibitor drug, to understand whether they are capable of binding to both COX-1 and P2Y12. Hence, this study was conducted to identify P2Y12 inhibitor drugs that have the ability to bind to COX-1, allowing them to be proposed as MAPT. Materials and methods: Molecular docking was employed to assess binding affinity, interaction types, amino acid residues, binding distances, and visualizations in both 3D and 2D formats. The applications utilized were BIOVIA Discovery Studio and AutoDock, while the websites utilized were research collaboratory for structural bioinformatics protein data bank (RCSB PDB) and PubChem. Results: In silico findings reveal differences in binding strength among clopidogrel, prasugrel, and ticagrelor to COX-1 and P2Y12, with ticagrelor emerging as the stronger ligand due to a higher number of bindings and/or closer binding distances. Notably, only prasugrel and ticagrelor demonstrate the ability to bind to the active site of COX-1. Conclusion: Therefore, prasugrel and ticagrelor emerge as potential MAPT agents for ACS patients. Keywords: clopidogrel, prasugrel, ticagrelor, antiplatelet, ACS, in silico, molecular docking

The rapid aging of the global population is a major worldwide issue because of the close relationship between age and the development of several diseases. Aging or senescence is among the most widely studied topics at the moment. However, no pharmaceuticals have been developed that claim to possess anti-senescence properties. Andrographis paniculata, is a medicinal plant found widely throughout tropical and subtropical Asia. This review aims to identify the potential anti- senescence effect of A. paniculata extract and its bioactive compounds. By following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, five databases were used and in vivo and in vitro studies were included in this review. A. paniculata extracts and their bioactive compounds exert anti-senescence properties through their anti-inflammatory and antioxidant properties. This herb and its compounds enhanced memory, cognitive function and behaviour in Alzheimer's disease. The extract also promoted cell cycle progression and proliferation in the skin. In addition, andrographolide exhibited anti-senescence effects in endothelial cells through the activation of PI3K/Akt/Nrf and PI3K/Akt/AP-1 pathways. A. paniculata along with its bioactive compounds including andrographolide and 14-deoxyandrographolide, may have the potential to be used as anti-senescence through anti-inflammatory and antioxidant properties. However, the specific markers to evaluate the senescence are necessary to be conducted. Any clinical trials should be done to establish these findings. Since in clinical settings this potential herbal may be used for long-life time, the safety profile and toxicity of A. paniculata should be considered. Keywords: herbal plants, Andrographis paniculata, andrographolide, bioactive compounds, senescence

Myocardial infarction is one of the leading causes of death worldwide. Current treatments do not compensate for the loss of cardiomyocytes, thus progression to heart failure is often inevitable. In myocardial infarction, the occlusion of coronary arteries and sudden restoration of blood flow give rise to ischemia-reperfusion injury, which leads to cardiomyocyte death. Mitochondria are not only involved in the bioenergetic aspect of the heart but also play a pivotal role in cell death during ischemia-reperfusion injury. Their morphology dynamically changes via fusion and fission in a balanced manner to maintain cellular health. However, ischemia-reperfusion injury triggers excessive mitochondrial fission, which is pathological to the myocardium. This review article discusses the association between myocardial ischemia-reperfusion injury and mitochondrial dynamics, serving as a rationale for a novel therapeutic strategy for myocardial infarction. Strategic modulation of mitochondrial dynamics under this pathological setting has been shown to be effective for cardioprotection. Increasing mitochondrial fusion or reducing excessive mitochondrial fission in the myocardial tissue could prevent cardiomyocyte death, thereby reducing infarct size. Proof-of-concept studies have utilized small molecules and peptides to implement this strategy into in vivo myocardial ischemia-reperfusion injury models. However, there remains a need to address the issues of specificity, bioavailability, and potency of these pharmacological agents before future application in cardiovascular therapeutics. Nevertheless, there has been growing interest in this therapeutic strategy in recent years, rendering it an attractive approach for ischemia-reperfusion injury in the heart. Keywords: mitochondria, heart, ischemia-reperfusion, cardioprotection

Background: Obesity is a global epidemic caused by excessive body fat, which is increasing free fatty acids in the liver, causing oxidative stress and liver cell damage. Green tea yogurt (GTY) with encapsulated Lacticaseibacillus paracasei E1 (GTY-LpE1) might have a beneficial effect in reducing liver cell damage. This study was conducted determine GTY-LpE1 effect on superoxide dismutase (SOD) expression, malondialdehyde (MDA) expression and liver histopathology in high-fat high-fructose diet (HFFD) mice. Material and Methods: A completely randomized design (CRD) with 7 groups, including normal diet (ND) group, HFFD group, 1.3 mg/kg BW simvastatin (SIM)-administered HFFD group, 5 g/kg BW probiotic yoghurt (PY)-administered HFFD (PY), 2.5 g/kg BW GTY-administered HFFD (2.5 GTY), 5 g/kg BW GTY-administered HFFD (5 GTY), and 10 g/kg BW GTY-administered HFFD (10 GTY). The diet was given for 16 weeks, followed by oral administration of sim/yoghurt during the last 4 weeks. Mice were sacrificed and the liver was collected. SOD and MDA expression were analyzed by flow cytometry. Histopathology analysis was done by evaluating hematoxylin-eosin (HE) staining of the liver. Result: The percentage of necrotic cells were 34.55, 34.31, and 21.95%, when treated with 2.5, 5, and 10 g/kg BW with GTY-administered HFFD, respectively, these were lower than the ones in the HFFD group (69.49%). The percentage of MDA expression were 15.55, 18.69, and 22.42%, respectively, these were lower than the ones in the HFFD group as well. The percentage of SOD expression were 9.49, 7.85, and 11.11%, respectively, these were higher than the ones in the HFFD group (3.44%). Conclusion: GTY-LpE1 could decrease the number of necrotic cells in the HFFD mice livers and improve the hepatocyte damage by reducing MDA expression and enhancing SOD expression. GTY-LpE1 can be used as an alternative food to control obesity. Keywords: alginate, chitosan, encapsulation, green tea, probiotic

Background: Alzheimer's disease (AD) is a neurodegenerative disorder with a significant burden on public health, and current treatments offer limited efficacy. This study investigated the effectiveness of adipose-derived mesenchymal stem cells (AD-MSCs) on the expression of the nestin gene and amyloid plaque in an aluminum chloride (AlCl3)-driven Alzheimer's rat model. Materials and methods: AD-MSCs were characterized using flow cytometry. Adult male Wistar rats were treated with/without AlCl3 and injected with/without AD-MSCs. After 5 days of AlCl3 ingestion and 4 weeks of subsequent AD-MSCs intraperitoneal injection, behavioral and molecular assessments were conducted. The Y-maze alternation test was used to test spatial learning of rats. Nestin gene expression was evaluated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The presence of amyloid plaque in the cortex and the hippocampus was evaluated through Congo red staining. Results: AD-MSC-like cells expressed the MSC markers CD90, CD73 and CD105. The Y-maze alternation result for rats treated with AlCl3 and AD-MSC-like cells was significantly higher compared with rats treated with AlCl3 only. Nestin gene expression was significantly higher in rats treated with AlCl3 and AD-MSC-like cells compared to those treated with AlCl3 only. After AD-MSC-like cells treatment, the Congo red staining results of rat’s cortex and hippocampus were significantly decreased. Conclusion: The findings suggest that AD-MSC-like cells possess therapeutic potential in restoring neural plasticity, amyloid plaque clearance and warrant further investigation for AD treatment. This study contributes to the emerging field of stem cell therapy for neurodegenerative diseases by highlighting the promise of AD-MSCs. Keywords: Alzheimer's disease, adipose-derived mesenchymal stem cells, neural plasticity, congo red staining, stem cell therapy

Background: Early onset preeclampsia (PE) is considered a fetal disorder that is associated with placental dysfunction. While late onset preeclampsia is considered a maternal disorder that associated with a normal placenta. An imbalance of angiogenesis factors, namely soluble Fms-like tyrosine kinase 1 (sFlt-1) and placental growth factor (PlGF) is not only a strong predictor of PE but can also be a predictor of poor outcomes or adverse complications of PE. This study was conducted to analyze the difference between angiogenesis factor sFlt-1 and PlGF in maternal serum between patients with early and late onset PE. Material and Methods: This was a cross-sectional study involving pregnant women with PE who were ≥18 years old and gestational age >20 weeks had singleton pregnancies. Subjects who had a major morphological abnormality of a fetus diagnosed with USG and chromosomal abnormality of the fetus were be excluded. Systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) were measured using a digital blood pressure measuring instrument. sFlt-1 and PlGF levels were measured using enzyme linked immunosorbent assay (ELISA). Result: This study showed the level of maternal serum sFlt-1 was 7522.95 pg/mL and the level of maternal serum PlGF was 222.25 pg/mL. There was a difference between sflt-1, PlGF, and the sflt-1/PlGF ratio (p=0.00; p=0.00; p=0.00) in early and late onset of PE where at early onset PE was found higher sflt-1 and late-onset PE had lower PlGF and higher sflt-1/PlGF ratio. Conclusion: There are differences in sFlt-1, PlGF, and sFlt-1/PlGF in early and late onset PE, as higher sflt-1 is found in early onset preeclampsia, while lower PlGF and higher sflt-1/PlGF ratio are found in late onset preeclampsia. Keywords: early onset, late onset, preeclampsia, maternal angiogenesis factor, Flt-1, PlGF

Background: Cellular senescence and the senescence-associated secretory phenotype (SASP) are pivotal factors influencing aging and age-related diseases. SASP secretes cytokines, chemokines, metalloproteinases, and growth factors that cause chronic inflammation. C-C ligand 2 (CCL2) and transforming growth factor-beta (TGF-β) are SASP markers secreted by senescent cells. This study investigated the relationship between the FOXO3 variant rs2802292 and SASP markers, focusing on CCL2 and TGF-β. Materials and methods: A cross-sectional study involving 72 elderly individuals from Jakarta was conducted. A sandwich enzyme-linked immunosorbent assay (ELISA)was used to quantify CCL2 and TGF-β concentrations. Random blood glucose, blood pressure, and FOXO3 rs2802292 genotyping data were obtained from a previous study. Differences in CCL2 and TGF-β concentrations between genotype groups were analyzed using one-way ANOVA and the Kruskal-Wallis test. Meanwhile, differences in CCL2 and TGF-β concentrations between allele groups were analyzed using the Mann-Whitney test. Results: The CCL2 and TGF-β concentrations of the subjects were 66.5 (10.58-190.9) pg/mL and 6,319 (2,379-13,846) pg/mL, respectively. There were significant differences in CCL2 concentrations among the FOXO3 rs2802292 genotypes (p=0.041). However, there were no significant differences in TGF-β concentrations among FOXO3 rs2802292 genotypes (p=0.955). Subjects with the G allele had significantly lower CCL2 concentrations compared with those with the T allele (p=0.033). TGF-β concentrations did not significantly differ between G and T alleles (p=0.771). Conclusion: CCL2 concentrations are associated with the FOXO3 variant rs2802292 in the elderly population. The T allele of FOXO3 rs2802292 increased CCL2 concentration and slightly decreased TGF-β concentration in elderly individuals. Keywords: aging, SASP, CCL2, TGF-β, SNP, FOXO3, rs2802292

Cavitation in water only, no matter whether hydrodynamic or acoustic, is a zero-waste circular economy process to convert industrial citrus processing waste into high-performance polysaccharides in high demand in a single-step at room temperature and ambient pressure using a modest amount of electricity as the only energy input. Following previous reports in which we used hydrodynamic cavitation, we now use an industrial acoustic sonicator to demonstrate the general viability of cavitation to convert biowaste residue of the industrial squeezing of pigmented sweet orange (Citrus sinensis) into highly bioactive “IntegroPectin” pectin and micronized cellulose “CytroCell”. From biomedicine through advanced composite membranes, said biomaterials hold great applicative potential. We conclude discussing the economic and technical feasibility of industrial implementation of the “CytroCav” process.

A multiscale methodology approach was employed integrating microscopic analysis of the biomasses present in the biocomposite (lignocellulosic and fungal) to understand their macroscopic response in terms of physical and mechanical properties. Colombian native strain Ganoderma gibbosum, used for the first time in the production of biocomposites was cultivated on peach palm fruit peel flour and sugar cane bagasse wet dust, individually and as a mixture. During the solid-state fermentation were monitoring the change that occurred in substrate composition such as glucan, arabinoxylan, and lignin through biomass compositional analysis using structural carbohydrates and lignin. Moreover, fungal biomass formation was monitored via scanning electron microscopy. The resulting biocomposites underwent characterization through flexural and water absorption tests. Our findings indicated that G. gibbosum primarily degraded the polysaccharides in each of the evaluated media. However, lignin degradation to 15.06 g/g was only observed in the mixture biocomposite of peach palm fruit peel fluor and sugarcane bagasse wet dust in a ratio of 1꞉1, accompanied by a reduction in glucan and arabinoxylan weights to 26.1 and 7.72 g/g, respectively. This polymer degradation, combined with a protein-rich source in the mixture biocomposite of peach palm fruit peel fluor and sugarcane bagasse wet dust in a ratio of 1꞉1, facilitated the production of a fungal skin (biological matrix) with a high hyphal density of 65%, contributing to Young's modulus of 3.83 MPa, elongation without failure, and low water absorption rate in this biocomposite (55%). The lignocellulosic biomass in the culture media acted as a filler for mechanical interlocking with the matrix and provided attachment points for water absorption. Thus, our study establishes a connection between the microscopic scale and the macroscopic behavior of this biocomposite, assessing structural carbohydrates and lignin analysis during solid-state fermentation (SSF), laying the groundwork for a more customized design of mycelium-based biocomposites. Finally, this study demonstrates the possibility of tailoring nutrient composition by designing their culture media to obtain physical-mechanical properties according to the application requirement.

This study aimed to systematically find an alternative solvent to replace hexane for the extraction of bio-oil with high γ-oryzanol content from rice bran (RB). The selection involved predicting solubility through Hansen solubility theory, experimental validation, determination of suitable extraction conditions, and comparison of oil quality with that of conventional hexane. A wide variety of solvents: subcritical water (SCW), supercritical carbon dioxide (SCCO2), bio-based solvents (alcohols and terpenes), and liquefied dimethyl ether (LDME), were initially assessed for rice bran oil (RBO) and γ-oryzanol solubility using Hansen solubility spheres. Solvents demonstrating high solubility for both RBO and γ-oryzanol, including LDME, ethyl acetate, acetone, and others (alcohols and SCCO2) known for effective vegetable oil extraction, were selected/identified for experimental extraction comparison. Among these, LDME performed better overall, affording greater solubility and requiring less solvent, shorter duration, lower pressure, and no additional co-solvents for equivalent extractions. Optimal conditions for LDME extraction were identified as 30 °C with a solvent-to-sample ratio of 10 mL/g and an extraction time of 10 min. Oils extracted with LDME and hexane displayed similar fatty acid compositions and no adverse effects on RB protein and carbohydrate structures after LDME extraction were observed. This study demonstrates LDME as a promising alternative to replace hexane for RBO extraction to further valorize this abundant low-cost RB residue into bio-oil and its γ-oryzanol and de-oil RB co-products.

Lignin is a rich renewable aromatic resource that can produce high-value-added chemicals. Lignin is regarded as one of the three major components of lignocellulosic biomass, which is composed of phenylpropane units connected by CO bond and CC bond. The cleavage of two chemical bonds is the main catalytic pathway in the production of chemicals and fuels from lignin. Although the cleavage of CO converts lignin into valuable aromatic compounds and renewable carbon sources, selective depolymerization for CC bonds is an important method to increase the yield of aromatic monomers. Therefore, in this review, we summarized the latest research trends on CC bond selective cleavage in lignin and lignin model compounds, focusing on various catalytic systems, including hydrogenolysis, oxidate, photocatalysis, and electrocatalysis. By analyzing the current status of CC bond breakage, the core issues and challenges related to this process and the expectations for future research were emphasized.

Cellulose nanocrystals (CNCs) have garnered increased attention due to their renewable nature, abundant feedstock availbility, and good mechanical properties. However, one of the bottlenecks for its commercial production is the drying process. Because of the low CNC concentrations in suspension after isolation, CNC drying requires the removal of a large amount of water to obtain dry products for the following utilization and saving shipping costs. A novel multi-frequency, multimode, modulated ultrasonic drying technology was developed for CNC drying to improve product quality, reduce energy consumption, and increase production rate. CNCs dried with different drying technologies were characterized by Fourier transform infrared (FT-IR) spectra analysis, X-ray diffraction (XRD) analysis, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and redispersibility to measure the quality and property changes. Under the same temperature and airflow rate, ultrasonic drying enhanced drying rates, resulting in at least a 50% reduction in drying time compared to hot air drying. The mean particle sizes of CNC from ultrasonic drying changed little with settling time, indicating good redispersibility. In addition, ultrasonic dried CNCs exhibited good stability in aqueous solutions, with the zeta potentials ranging from –35 to –65 mV. Specific energy consumption and CO2 emissions of various CNC drying technologies were evaluated. Energy consumption of ultrasonic drying is significantly reduced compared to other drying technologies. Moreover, the potential CO2 emissions of the fully electrified ultrasonic drying could be net zero if renewable electricity is used.

Organotin mercaptide-based thermal stabilizer is recognized for its effectiveness in enhancing thermal stability of polyvinyl chloride (PVC). In this study, we synthesized an organotin mercaptide-based thermal stabilizer from palm fatty acid distillate, which is a by-product of palm oil refining process, and then evaluated its thermal stabilizing effects on PVC and compared its efficacy and economics to those of mixed metal stearate. The synthesized thermal stabilizer manifests as methyltin mercaptoethyl carboxylate sulfides. Both dehydrochlorination and two-roll mill discoloration tests have demonstrated the high efficacy of the resulting thermal stabilizer in stabilizing PVC, surpassing the performance of mixed metal stearate, as evidenced by the lower dosage required. The synthesized PVC thermal stabilizer not only provides effective stabilization but also presents a competitive viable alternative.

Caragana korshinskii kom. (CKK) waste, a common forestry byproduct in northwest of China, presents challenges in its transformation into alternative ruminant feed due to its initial nutritional limitations and unappealing palatability. Conventional strategies, such as ensiling and fungal-based solid-state fermentation (SSF) cannot effectively address this issue in practice. Herein, a two-stage bioaugmentation (TBA) process was devised, leveraging the benefits of ensiling and SSF. During the anaerobic ensiling phase, CKK waste was inoculated with Lactiplantibacillus plantarum LP1, effectively suppressing potential animal pathogens such as Aspergillus and Nocardiopsis while enriching the material with potential probiotics like Pediococcus and Lactiplantibacillus, reaching an abundance of 95.7%. In the subsequent aerobic SSF stage, the ensiled CKK underwent inoculation with the white-rot fungus Irpex lacteus F17, which became enriched to 87.9%. Comprehensive multi-omics analysis identified Irpex as the key taxon, possessing an extensive redox enzyme system that led to the improvement in nutrient composition, reduction of astringent phenolic substances, and mitigation of mycotoxins. As a result, the crude protein content of the CKK increased by 39.2%, while lignin, total phenolic substances, and tannic acid content decreased by 24.4%, 52.2%, and 51.4%, respectively. The mycotoxin levels, including aflatoxin B1, zearalenone, and vomitoxin, were rendered negligible, confirming the safety. Overall, this study demonstrates the TBA strategy can successfully transform challenging and unpalatable CKK waste into a nutrient-enriched and safe mycelium-based bioproduct, thereby enabling the valorization of a previously underutilized forestry resource as a promising alternative feed.

Selective conversion of biomass into targeted molecules like polyols, especially, from cellulosic compounds, is being widely investigated as a sustainable process to produce biodiesel and bio-additives. The known process involves two steps, namely hydrolysis and hydrogenation. Thus, it requires two different catalytic materials or bifunctional catalysts. In this context, the present work reports a new catalytic approach based on the use of tandem catalysts, consisting of the combination of an acid solid catalyst (active for hydrolysis) and a supported metal catalyst (active for hydrogenation). Two different functionalized activated carbons and the resin Amberlyst 15 have been tested as solid acid catalysts, and Ru nanoparticles supported on the original activated carbon (SA) are the metal catalyst part of the tandem. All the tested tandem catalysts exhibited higher activity than the supported Ru catalyst did. The highest cellulose conversion and selectivity to sorbitol (70% and 86%, respectively) have been obtained over a novel tandem catalyst, which resulted from a physical mixture between a sulfuric acid modified SA carbon (SASu) and Ru loaded SA (Ru/SA), leading to a tandem catalyst (Ru/SA+SASu). This novel-designed tandem catalyst is reusable. Based on tandem catalysts with a solid-solid system combination, the adopted novel-designed catalytic approach is cost-efficient and sustainable, and can be considered promising for the green production of high-added-value chemicals.

Bacterial cellulose is a versatile material with applications in many industries. However, the widespread uptake of bacterial cellulose faces challenges including high production costs and lack of scalability. One approach to address these obstacles is the use of alternative substrates and media, compared to the Hestrin–Schramm (HS) media. By evaluating and selecting appropriate media and substrates, the production of bacterial cellulose can be more efficient: enabling sustainable systems and supply chains where less energy and materials are lost, and the output production is increased. The purpose of this paper is to analyze the current landscape of bacterial cellulose alternative media and substrates (ingredients). Through a systematic review of 198 papers, this review identifies 299 alternative substrates from 12 industries and 101 bacterial cellulose-producing strains, which were systematically compared. This review also finds that there are methodological gaps in this field such as data variability, papers mislabelling the HS media or not using a comparison media, and a lack of strain names. This alternative substrate analysis for bacterial cellulose production demonstrates that overall, for some applications alternative substrates can be taken into consideration that are not only cheaper, but also produce higher yields than HS media.

Lignin serves as a promising Ultraviolet (UV) absorber within sunscreen industry. However, the commercial development of lignin-containing sunscreens faces challenges due to their low sun protection factor (SPF) and dark color in cosmetics industry. In this study, dual modifications on the chemical and physical structures of lignin were conducted to address these challenges. Initially, methylene bis-benzotriazolyl tetramethylbutylphenol (MBBT) was grafted onto alkali lignin (AL) through an atom transfer radical polymerization reaction, resulting in a polymer of AL-graft-MBBT3 (AL-g-MBBT3). The sunscreen prepared with 10% AL-g-MBBT3 displays outstanding sun protection performance with a SPF of 42.93 and a light color with a color difference value (ΔE) of 45.6, in contrast to 10% AL with a SPF of 4.74 and a ΔE value of 49.5. Subsequently, AL-g-MBBT3 was transformed into normal submicron spheres (AL-g-MBBT3 N) and TiO2-loading submicron spheres (AL-g-MBBT3/TiO2). The sun protection performances of 10% AL-g-MBBT3 N@C and AL-g-MBBT3/TiO2@C sunscreens obviously surpass that of AL-g-MBBT3@C sunscreen, achieving SPFs of 60.38 and 66.20, respectively. Additionally, there is a considerable improvement in the color of these sunscreens, with ΔE values of 41.8 and 36.3, respectively. These results provide valuable insights into exploring lignin's high-value applications in sunscreen.

As the global population continues growing, the demand for textiles also increases, putting pressure on cotton manufacturers to produce more natural fiber from this already undersupplied resource. Synthetic fibers such as polyester (PET) can be manufactured quickly and cheaply, but these petroleum-based products are detrimental to the environment. With increased efforts to encourage transparency and create a more circular textile economy, other natural alternatives must be considered. This article discusses the existing condition and future possibilities for man-made cellulosic fibers (MMCFs), with an emphasis on using non-woody alternative feedstocks as a starting material. This work focuses on conversion technology suitable for producing textile-grade fibers from non-wood-based dissolving pulp, which may be different in nature from its woody counterpart and therefore behave differently in spinning processes. Derivatization and dissolution methods are detailed, along with spinning techniques and parameters for these processes. Existing research related to the spinning of non-woody-based dissolving pulp is covered, along with suggestions for the most promising feedstock and technology combinations. In addition, an emerging method of conversion, in which textile fibers are spun from a hydrogel made of an undissolved nano/micro-fibrillated fiber suspension, is briefly discussed due to its unique potential. Methods and concepts compiled in this review relate to utilizing alternative feedstocks for future fibers while providing a better understanding of conventional and emerging fiber spinning processes for these fibers.

Many effective pretreatment methods (such as dilute acid, dilute alkali, ionic liquids, etc.) have been developed for lignocellulose upgrading, but several defaults of low working mass, high sugar loss and extra cost of solid-liquid separation and water washing hinder their large-scale application in industry. Besides, the valorization of lignin-rich residue from pretreated biomass after hydrolysis or fermentation greatly contributes to the economy and sustainability of lignocellulosic biorefinery, which is usually underestimated. This study developed a densification pretreatment with binary chemicals (densifying lignocellulosic biomass with sulfuric acid (SA) and metal salt (MS) followed by autoclave treatment ((DLCA(SA-MS)), which was conducted under mild condition (121 °C) with a biomass working mass as high as 400 kg/m3. The DLCA(SA-MS) biomass achieved over 95% sugar retention, 90% enzymatic sugar conversion and a high concentration of fermentable sugar (212.3 g/L) with superior fermentability. Furthermore, bio-adsorbent derived from DLCA(SA-MS) biomass residue was highly adsorptive and suitable for dyeing wastewater treatment, providing a feasible and eco-friendly method for lignin-rich residue valorization. These findings indicated that DLCA(SA-MS) pretreatment enables the full-component utilization of biomass and boosts the economic viability of lignocellulosic biorefinery.

Abstract Background The prevalence of age-related disorders, particularly in neurological and cardiovascular systems, is an increasing global health concern. Mesenchymal stem cell (MSC) therapy, particularly using human umbilical cord-derived MSCs (HUCMSCs), has shown promise in mitigating these disorders. This study investigates the effects of HUCMSCs on aging-related conditions in a senescence-accelerated mouse model (SAMP8), with a focus on DNA damage, gut microbiota alterations, and metabolic changes. Methods SAMP8 mice were treated with clinical-grade HUCMSCs via intraperitoneal injections. Behavioral and physical assessments were conducted to evaluate cognitive and motor functions. The Single-Strand Break Mapping at Nucleotide Genome Level (SSiNGLe) method was employed to assess DNA single-strand breaks (SSBs) across the genome, with particular attention to exonic regions and transcription start sites. Gut microbiota composition was analyzed using 16S rRNA sequencing, and carboxyl metabolomic profiling was performed to identify changes in circulating metabolites. Results HUCMSC treatment significantly improved motor coordination and reduced anxiety in SAMP8 mice. SSiNGLe analysis revealed a notable reduction in DNA SSBs in MSC-treated mice, especially in critical genomic regions, suggesting that HUCMSCs may mitigate age-related DNA damage. The functional annotation of the DNA breaktome indicated a potential link between reduced DNA damage and altered metabolic pathways. Additionally, beneficial alterations in gut microbiota were observed, including an increase in short-chain fatty acid (SCFA)-producing bacteria, which correlated with improved metabolic profiles. Conclusion The administration of HUCMSCs in SAMP8 mice not only reduces DNA damage but also induces favorable changes in gut microbiota and metabolism. The observed alterations in DNA break patterns, along with specific changes in microbiota and metabolic profiles, suggest that these could serve as potential biomarkers for evaluating the efficacy of HUCMSCs in treating age-related disorders. This highlights a promising avenue for the development of new therapeutic strategies that leverage these biomarkers, to enhance the effectiveness of HUCMSC-based treatments for aging-associated diseases.

Abstract Periodontitis, a globally prevalent chronic inflammatory disease is characterized by the progressive degradation of tooth-supporting structures, particularly the periodontal ligament (PDL), which can eventually result in tooth loss. Despite the various clinical interventions available, most focus on symptomatic relief and lack substantial evidence of supporting the functional regeneration of the PDL. Dental stem cells (DSCs), with their homology and mesenchymal stem cell (MSC) properties, have gained significant attention as a potential avenue for PDL regeneration. Consequently, multiple therapeutic strategies have been developed to enhance the efficacy of DSC-based treatments and improve clinical outcomes. This review examines the mechanisms by which DSCs and their derivatives promote PDL regeneration, and explores the diverse applications of exogenous implantation and endogenous regenerative technology (ERT) aimed at amplifying the regenerative capacity of endogenous DSCs. Additionally, the persistent challenges and controversies surrounding DSC therapies are discussed, alongside an evaluation of the limitations in current research on the underlying mechanisms and innovative applications of DSCs in PDL regeneration with the aim of providing new insights for future development. Periodontitis, a chronic inflammatory disease, represents a major global public health concern, affecting a significant proportion of the population and standing as the leading cause tooth loss in adults. The functional periodontal ligament (PDL) plays an indispensable role in maintaining periodontal health, as its structural and biological integrity is crucial for the long-term prognosis of periodontal tissues. It is widely recognized as the cornerstone of periodontal regeneration Despite the availability of various treatments, ranging from nonsurgical interventions to guided tissue regeneration (GTR) techniques, these methods have shown limited success in achieving meaningful PDL regeneration. As a result, the inability to fully restore PDL function underscores the urgent need for innovative therapeutic strategies at reconstructing this essential structure. Stem cell therapy, known for its regenerative and immunomodulatory potential, offers a promising approach for periodontal tissue repair. Their application marks a significant paradigm shift in the treatment of periodontal diseases, opening new avenues for functional PDL regeneration. However, much of the current research has primarily focused on the regeneration of alveolar bone and gingiva, as these hard and soft tissues can be more easily evaluated through visual assessment. The complexity of PDL structure, coupled with the intricate interactions among cellular and molecular components, presents significant scientific and clinical hurdles in translating DSC research into practical therapeutic applications. This review provides a thorough exploration of DSC dynamics in periodontal regeneration, detailing their origins, properties, and derived products, while also examining their potential mechanisms and applications in PDL regeneration. It offers an in-depth analysis of the current research, landscape, acknowledging both the progress made and the challenges that remain in bridging the gap between laboratory findings and clinical implementation. Finally, the need for continued investigation into the intricate mechanisms governing DSC behavior and the optimization of their use in regenerative therapies for periodontal diseases is also emphasized.

Abstract Endothelial-mesenchymal transformation (EndoMT) is the process through which endothelial cells transform into mesenchymal cells, affecting their morphology, gene expression, and function. EndoMT is a potential risk factor for cardiovascular and cerebrovascular diseases, tumor metastasis, and fibrosis. Recent research has highlighted the role of exosomes, a mode of cellular communication, in the regulation of EndoMT. Exosomes from diseased tissues and microenvironments can promote EndoMT, increase endothelial permeability, and compromise the vascular barrier. Conversely, exosomes derived from stem cells or progenitor cells can inhibit the EndoMT process and preserve endothelial function. By modifying exosome membranes or contents, we can harness the advantages of exosomes as carriers, enhancing their targeting and ability to inhibit EndoMT. This review aims to systematically summarize the regulation of EndoMT by exosomes in different disease contexts and provide effective strategies for exosome-based EndoMT intervention.

Abstract Hippo pathway is an evolutionarily conservative key pathway that regulates organ size and tissue regeneration by regulating cell proliferation, differentiation and apoptosis. Yes-associated protein 1 (YAP)/ WW domain-containing transcription regulator 1 (TAZ) serves as a pivotal transcription factor within the Hippo signaling pathway, which undergoes negative regulation by the Hippo pathway. The expression of YAP/TAZ affects various biological processes, including differentiation of osteoblasts (OB) and osteoclasts (OC), cartilage homeostasis, skeletal muscle development, regeneration and quality maintenance. At the same time, the dysregulation of the Hippo pathway can concurrently contribute to the development of various musculoskeletal disorders, including bone tumors, osteoporosis (OP), osteoarthritis (OA), intervertebral disc degeneration (IDD), muscular dystrophy, and rhabdomyosarcoma (RMS). Therefore, targeting the Hippo pathway has emerged as a promising therapeutic strategy for the treatment of musculoskeletal disorders. The focus of this review is to elucidate the mechanisms by which the Hippo pathway maintains homeostasis in bone, cartilage, and skeletal muscle, while also providing a comprehensive summary of the pivotal role played by core components of this pathway in musculoskeletal diseases. The efficacy and feasibility of Hippo pathway-related drugs for targeted therapy of musculoskeletal diseases are also discussed in our study. These endeavors offer novel insights into the application of Hippo signaling in musculoskeletal disorders, providing effective therapeutic targets and potential drug candidates for treating such conditions.

Abstract Background Mesenchymal stem cells (MSCs) have emerged as promising candidates for immune modulation in various diseases that are associated with dysregulated immune responses like Graft-versus-Host-Disease (GVHD). MSCs are pleiotropic and the fate of MSCs following administration is a major determinant of their therapeutic efficacy. Methods Human MSCs were derived from bone marrow (BM) and Wharton’s Jelly (WJ) and preconditioned through exposure to hypoxia and induction of apoptosis, either sequentially or simultaneously. The immune programming potential of preconditioned MSCs was evaluated by assessing their effects on T cell proliferation, induction of Tregs, programming of effector T-cell towards Th2 phenotype, macrophage polarization in the direct co-culture of MSCs and aGVHD patients-derived PBMNCs. Additionally, efferocytosis of MSCs and relative change in the expression of immunomodulatory soluble factors were examined. Results Our study demonstrated that hypoxia preconditioned apoptotic MSCs (BM-MSCs, WJ-MSCs) bear more immune programming ability in a cellular model of acute Graft-versus-Host-Disease (aGVHD). Our findings revealed that WJ-MSCsHYP+APO were superior to BM-MSCsHYP+APO for immune regulation. These induced the differentiation of CD4+T-cell into Tregs, enhanced Th2 effector responses, and simultaneously mitigated Th1- and Th17 responses. Additionally, this approach led to the polarization of M1 macrophages toward an M2 phenotype. Conclusion Our study highlights the potential of WJ-MSCs conditioned with hypoxia and apoptosis concurrently, as a promising therapeutic strategy for aGVHD. It underscores the importance of considering MSC apoptosis in optimizing MSCs-based cellular therapy protocols for enhanced therapeutic efficacy in aGvHD. Graphical Abstract

Abstract Background Critical limb ischemia (CLI) is a condition characterized by insufficient blood flow to the lower limbs, resulting in severe ischemia and potentially leading to amputation. This study aims to identify novel vasculogenic precursor cells (VPCs) in human bone marrow and evaluate their efficacy in combination with bone marrow-derived mesenchymal stem cells (BM-MSCs) for the treatment of CLI. Methods Ex vivo cultured VPCs and BM-MSCs from bone marrow were characterized and their effects on neovascularization and long-term tissue regeneration were tested in a mouse CLI model. Results VPCs, expressing high levels of hepatocyte growth factor and c-MET, were identified from human bone marrow aspirates. These cells exhibited strong vasculogenic capacity in vitro but possessed a cellular phenotype distinct from those of previously reported endothelial precursor cells in circulation or cord blood. They also expressed most surface markers of BM-MSCs and demonstrated multipotent differentiation ability. Screening of 376 surface markers revealed that VPCs uniquely display CD141 (thrombomodulin). CD141+VPCs are present in BM aspirates as a rare population and can be expanded ex vivo with a population doubling time of approximately 20 h, generating an elaborate vascular network even under angiogenic factor-deficient conditions and recruiting BM-MSCs to the network as pericyte-like cells. Intramuscular transplantation of a combination of human CD141+VPCs and BM-MSCs at a ratio of 2:1 resulted in limb salvage, blood flow recovery, and regeneration of large vessels in the femoral artery-removed CLI model, with an efficacy superior to that of singular transplantation. Importantly, large arteries and arterioles in dual cell transplantation expressed human CD31 in the intima and human α-smooth muscle actin in media layer at 4 and 12 weeks, likely indicating their lineage commitment to endothelial cells and vascular smooth muscle, respectively, in vivo. Conclusion Dual-cell therapy using BM-derived CD141+ VPCs and BM-MSCs holds potential for further development in clinical trials to treat peripheral artery disease and diabetic ulcers.

Abstract Background Mesenchymal stem/stromal cells (MSCs) have attracted interest as a potential therapy given their anti-inflammatory and immunomodulatory properties. However, clinical trials using MSCs for acute respiratory distress syndrome (ARDS) have produced mixed and inconclusive data. In previous work, we performed a “head-to-head” comparison between different sources of MSCs and showed that each source had a unique genomic and proteomic “signature”. Method This study investigated which sources of MSC: bone marrow derived-MSCs (BM-MSCs), adipose tissue derived-MSCs (AD-MSCs) and umbilical cord derived-MSCs (UC-MSCs) would be the optimal candidate to be used as a therapy in an LPS-induced mouse model of ARDS. Immune cells assessment, tissue transcriptomics, animal survival, and endothelial-epithelial barrier assessment were used to evaluate their effects. Results When comparing the three most commonly used MSC sources, we found that UC-MSCs exhibited greater efficacy compared to other MSCs in improving animal survival, mitigating epithelial/endothelial damage, decreasing lung inflammation via reducing neutrophil infiltration, T cell proliferation, and M1 polarization. Bulk RNA sequencing of lung tissue also showed that UC-MSCs have the capability to downregulate extracellular trap formation, by the downregulation of key genes like Elane and Padi4. Notably, treatment with UC-MSCs demonstrated a significant reduction in Fc-γ R mediated phagocytosis, which has been associated with monocyte pyroptosis and intense inflammation in the context of COVID-19. Conclusion Our findings suggest that UC-MSCs are an optimal source of MSC to treat acute inflammatory conditions in the lungs, such as ARDS.

Abstract Background Hematopoietic stem cell transplantation (HSCT) is a curative treatment for many diverse blood and immune diseases. However, HSCT regimens currently commonly utilize genotoxic chemotherapy and/or total body irradiation (TBI) conditioning which causes significant morbidity and mortality through inducing broad tissue damage triggering infections, graft vs. host disease, infertility, and secondary cancers. We previously demonstrated that targeted monoclonal antibody (mAb)-based HSC depletion with anti(α)-CD117 mAbs could be an effective alternative conditioning approach for HSCT without toxicity in severe combined immunodeficiency (SCID) mouse models, which has prompted parallel clinical αCD117 mAbs to be developed and tested as conditioning agents in clinical trials starting with treatment of patients with SCID. Subsequent efforts have built upon this work to develop various combination approaches, though none are optimal and how any of these mAbs fully function is unknown. Methods To improve efficacy of mAb-based conditioning as a stand-alone conditioning approach for all HSCT settings, it is critical to understand the mechanistic action of αCD117 mAbs on HSCs. Here, we compare the antagonistic properties of αCD117 mAb clones including ACK2, 2B8, and 3C11 as well as ACK2 fragments in vitro and in vivo in both SCID and wildtype (WT) mouse models. Further, to augment efficacy, combination regimens were also explored. Results We confirm that only ACK2 inhibits SCF binding fully and prevents HSC proliferation in vitro. Further, we verify that this corresponds to HSC depletion in vivo and donor engraftment post HSCT in SCID mice. We also show that SCF-blocking αCD117 mAb fragment derivatives retain similar HSC depletion capacity with enhanced engraftment post HSCT in SCID settings, but only full αCD117 mAb ACK2 in combination with αCD47 mAb enables enhanced donor HSC engraftment in WT settings, highlighting that the Fc region is not required for single-agent efficacy in SCID settings but is required in immunocompetent settings. This combination was the only non-genotoxic conditioning approach that enabled robust donor engraftment post HSCT in WT mice. Conclusion These findings shed new insights into the mechanism of αCD117 mAb-mediated HSC depletion. Further, they highlight multiple approaches for efficacy in SCID settings and optimal combinations for WT settings. This work is likely to aid in the development of clinical non-genotoxic HSCT conditioning approaches that could benefit millions of people world-wide.

Abstract Background Rheumatoid arthritis (RA) is a chronic inflammatory disease that affects not only the joints but also has significant cardiovascular (CV) manifestations. The mechanistic interplay between RA and cardiovascular complications is not yet well understood due to the lack of relevant in vitro models. In this study, we established RA cardiac microtisses (cMTs) from iPSC-derived cardiomyocytes (CMs), endothelial cells (ECs) and cardiac fibroblasts (CFs) to investigate whether this fully human 3D multicellular system could serve as a platform to elucidate the connection between RA and CV disorders. Methods PBMC and FLS from healthy and RA donors were reprogrammed to hiPSCs with Sendai vectors. hiPSCs pluripotency was assessed by IF, FACS, spontaneous embryoid bodies formation and teratoma assay. hiPSCs were differentiated to cardiac derivatives such as CMs, ECs and CFs, followed by cell markers characterizations (IF, FACS, qRT-PCR) and functional assessments. 3D cMTs were generated by aggregation of 70% CMs, 15% ECs and 15% CFs. After 21 days in culture, structural and metabolic properties of 3D cMTs were examined by IF, qRT-PCR and Seahorse bioanalyzer. Results hiPSCs demonstrated typical colony-like morphology, normal karyotype, presence of pluripotency markers, and ability to differentiate into cells originating from all three germ layers. hiPSC-CMs showed spontaneous beating and expression of cardiac markers (cTnT, MYL7, NKX2.5, MYH7). hiPSC-ECs formed sprouting spheres and tubes and expressed CD31 and CD144. hiPSC-CFs presented spindle-shaped morphology and expression of vimentin, collagen 1 and DDR2. Self-aggregation of CMs/ECs/CFs allowed development of contracting 3D cMTs, demonstrating spherical organization of the cells, which partially resembled the cardiac muscle, both in structure and function. IF analysis confirmed the expression of cTnT, CD31, CD144 and DDR2 in generated 3D cMTs. RA cMTs exhibited significantly greater formation of capillary-like structures, mimicking enhanced vascularization—key RA feature—compared to control cMTs. Seahorse examination of cMTs revealed changes in mitochondrial and glycolytic rates in the presence of metabolic substrates and inhibitors. Conclusions The cMTs model may represent an advanced human stem cell-based platform for modeling CV complications in RA. The highly developed capillary-like structures observed within RA cMTs highlight a critical feature of inflammation-induced CV dysfunction in chronic inflammatory diseases.

Abstract Diabetes mellitus (DM), a chronic metabolic disease, poses a significant global health challenge, with current treatments often fail to prevent the long-term disease complications. Mesenchymal stem/stromal cells (MSCs) are, adult progenitors, able to repair injured tissues, exhibiting regenerative effects and immunoregulatory and anti-inflammatory responses, so they have been emerged as a promising therapeutic approach in many immune-related and inflammatory diseases. This review summarizes the therapeutic mechanisms and outcomes of MSCs, derived from different human tissue sources (hMSCs), in the context of DM type 1 and type 2. Animal model studies and clinical trials indicate that hMSCs can facilitate pleiotropic actions in the diabetic milieu for improved metabolic indices. In addition to modulating abnormally active immune system, hMSCs can ameliorate peripheral insulin resistance, halt beta-cell destruction, preserve residual beta-cell mass, promote beta-cell regeneration and insulin production, support islet grafts, and correct lipid metabolism. Moreover, hMSC-free derivatives, importantly extracellular vesicles, have shown potent experimental anti-diabetic efficacy. Moreover, the review discusses the diverse priming strategies that are introduced to enhance the preclinical anti-diabetic actions of hMSCs. Such strategies are recommended to restore the characteristics and functions of MSCs isolated from patients with DM for autologous implications. Finally, limitations and merits for the wide spread clinical applications of MSCs in DM such as the challenge of autologous versus allogeneic MSCs, the optimal MSC tissue source and administration route, the necessity of larger clinical trials for longer evaluation duration to assess safety concerns, are briefly presented. Graphical Abstract

Abstract Retinal degenerative diseases are a leading cause of vision loss and blindness globally, impacting millions. These diseases result from progressive damage to retinal pigment epithelial (RPE) cells for which no curative or palliative treatments exist. Cell therapy, particularly RPE transplantation, has emerged as a promising strategy for vision restoration. This review provides a comprehensive overview of the recent advancements in clinical trials related to RPE transplantation. We discuss scaffold-free and scaffold-based approaches, including RPE cell suspensions and pre-organized RPE monolayers on biomaterial scaffolds. Key considerations, such as the form and preparation of RPE implants, delivery devices, strategies, and biodegradability of scaffolds, are examined. The article also explores the challenges and opportunities in RPE scaffold development, emphasising the crucial need for functional integration, immunomodulation, and long-term biocompatibility to ensure therapeutic efficacy. We also highlight ongoing efforts to optimise RPE transplantation methods and their potential to address retinal degenerative diseases.

Abstract Background Endometriosis, due to its ambiguous symptoms, still remains one of the most difficult female diseases to treat, with an average diagnosis time of 7–9 years. The changing level of hypoxia plays an important role in a healthy endometrium during menstruation and an elevated expression of the hypoxia-inducible factor 1-alpha (HIF-1α) has been demonstrated in ectopic endometria. HIF-1α mediates the induction of proangiogenic factors and the development of angiogenesis is a critical step in the establishment and pathogenesis of endometriosis. Although the inhibition of angiogenesis has been proposed as one of the actionable therapeutic modalities, vascular normalization and re-oxygenation may become a possible new approach for therapeutic intervention. Methods Our goal was to investigate whether a selected murine model of endometriosis would be suitable for future studies on new methods for treating endometriosis. Non-invasive, high-resolution ultrasound-monitored observation was selected as the preclinical approach to obtain imaging of the presence and volume of the endometriotic-like lesions. The EF5 (2-(2-Nitro-1H-imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl)acetamide) compound that selectively binds to reduced proteins in hypoxic cells was used for hypoxia detection. The expression of Pten and other crucial genes linking endometriosis and hypoxia were also assessed. Results Using EF5, a pentafluorinated derivative of the 2-nitroimidazole that is metabolically reduced by oxygen-inhibitable nitroreductase, we confirmed that hypoxia did develop in the selected model and was detected in uterine and ectopic endometriotic lesions. Moreover, the changes in oxygen tension also influenced the expression level of significant genes related to endometriosis, like Pten, Trp53, Hif1a, Epas1, and Vegfa. Their strong modulation evidenced here is indicative of model reliability. Using high-resolution ultrasound-based imaging, we present a non-invasive method of visualization that enables the detection and observation of lesion evolution throughout the duration of the experiment, which is fundamental for further preclinical studies and treatment evaluation. Conclusions The selected model and method of visualization appear to be suitable for the study of new treatment strategies based on hypoxia alleviation and blood flow restoration.

Abstract Osteoblasts are mainly derived from mesenchymal stem cells in the bone marrow. These stem cells can differentiate into osteoblasts, which have the functions of secreting bone matrix, promoting bone formation, and participating in bone remodeling. The abnormality of osteoblasts can cause a variety of bone-related diseases, including osteoporosis, delayed fracture healing, and skeletal deformities. In recent years, with the side effects caused by the application of PTH drugs, biphosphonate drugs, and calmodulin drugs, people have carried out more in-depth research on the mechanism of osteoblast differentiation, and are actively looking for natural compounds for the treatment of osteoporosis. The Wnt/β-catenin signaling pathway is considered to be one of the important pathways of osteoblast differentiation, and has become an important target for the treatment of osteoporosis. The Wnt/β-catenin signaling pathway, whether its activation is enhanced or its expression is weakened, will cause a variety of diseases including tumors. This review will summarize the effect of Wnt/β-catenin signaling pathway on osteoblast differentiation and the correlation between the related proteins in the pathway and human diseases. At the same time, the latest research progress of natural compounds targeting Wnt/β-catenin signaling pathway against osteoporosis is summarized.

Abstract Background Esophageal Squamous Cell Carcinoma (ESCC) remains a predominant health concern in the world, characterized by high prevalence and mortality rates. Advances in single-cell transcriptomics have revolutionized cancer research by enabling a precise dissection of cellular and molecular diversity within tumors. Objective This study aims to elucidate the cellular dynamics and molecular mechanisms in ESCC, focusing on the transcriptional influence of STAT3 (Signal Transducer and Activator of Transcription 3) and its interaction with LHPP, thereby uncovering potential therapeutic targets. Methods Single-cell RNA sequencing was employed to analyze 44,206 cells from tumor and adjacent normal tissues of ESCC patients, identifying distinct cell types and their transcriptional shifts. We conducted differential gene expression analysis to assess changes within the tumor microenvironment (TME). Validation of key regulatory interactions was performed using qPCR in a cohort of 21 ESCC patients and further substantiated through experimental assays in ESCC cell lines. Results The study revealed critical alterations in cell composition and gene expression across identified cell populations, with a notable shift towards pro-tumorigenic states. A significant regulatory influence of STAT3 on LHPP was discovered, establishing a novel aspect of ESCC pathogenesis. Elevated levels of STAT3 and suppressed LHPP expression were validated in clinical samples. Functional assays confirmed that STAT3 directly represses LHPP at the promoter level, and disruption of this interaction by promoter mutations diminished STAT3's repressive effect. Conclusion This investigation underscores the central role of STAT3 as a regulator in ESCC, directly impacting LHPP expression and suggesting a regulatory loop crucial for tumor behavior. The insights gained from our comprehensive cellular and molecular analysis offer a deeper understanding of the dynamics within the ESCC microenvironment. These findings pave the way for targeted therapeutic interventions focusing on the STAT3-LHPP axis, providing a strategic approach to improve ESCC management and prognosis.

Abstract To elucidate the expression profiles, methylation states, and clinicopathological significance of the PRDM gene family, focusing on the MECOM gene's role in uterine endometrial cancer (UCEC) and its molecular interactions with the TGF-beta signaling pathway. Our methodology combined detailed bioinformatics analyses using UALCAN and GEPIA with in vitro assessments in HEC-1-A cells. Techniques included CRISPR-Cas9 for gene editing and various cellular assays (CCK-8, flow cytometry, Transwell) to evaluate the effects of MECOM on cell proliferation, migration, and apoptosis, alongside Western blot analysis for protein regulation in the TGF-beta pathway. MECOM was upregulated in UCEC tissues, influencing tumor cell behavior significantly. Knockout studies demonstrated reduced proliferation and migration and increased apoptosis, while overexpression showed reverse effects. Mechanistically, MECOM modulated critical proteins within the TGF-beta pathway, impacting cell cycle dynamics and apoptotic processes. The PRDM gene family, particularly MECOM, plays a crucial role in the pathogenesis and progression of UCEC, suggesting its utility as a target for novel therapeutic interventions. Our findings offer valuable insights for future research and potential clinical application in managing uterine endometrial cancer.

Abstract Background Premature ovarian insufficiency (POI) is an immune-related condition. Dihydroberberine (dhBBR) plays a regulatory role in maintaining the T-helper 17 (Th17)/regulatory T (Treg) cell balance. This study aimed to explore the action mechanisms of dhBBR on POI. Methods In vivo, female BALB/c mice were used as POI models, treated with dhBBR, or injected with recombinant interleukin (rIL)-17 and anti-CD25 monoclonal antibody. Hematoxylin and eosin staining was used to validate the model and assess the therapeutic effects of dhBBR. mRNA expression levels of cytochrome P450 (Cyp)-17a1, Cyp19a1, Cyp11a1, steroidogenic acute regulatory protein, and luteinizing hormone receptor in mouse ovaries were quantified via quantitative polymerase chain reaction (qPCR). Enzyme-linked immunosorbent assay was used to determine the cytokine and sex hormone levels. Immunohistochemical staining for cleaved-caspase 3 and Ki-67 were performed to assess ovarian cell apoptosis and proliferation. Flow cytometry was used to analyze the Th17/Treg cell balance in the ovary and spleen. In vitro cytotoxicity of dhBBR was measured using the cell counting kit-8 assay. GTP-Ras homolog enriched in brain (Rheb) activity was determined via immunofluorescence assay. Co-immunoprecipitation was performed to assess Rheb activity, Th17 or Treg induction, and binding between Rheb and mammalian target of rapamycin (mTOR) after dhBBR treatment. Flow cytometry and qPCR assays were used to verify the effect of dhBBR on CD4 + cell differentiation. Finally, Rheb/mTOR pathway activation was confirmed via western blotting of proteins, including mTOR, p-mTOR, p70S6K, p-p70S6K, 4E-BP1, and p-4E-BP1. Results dhBBR improved the ovarian function in a dose-dependent manner. It also decreased ovarian cell apoptosis and increased cell proliferation. It decreased Th1 and Th17 cell proportions but increased Treg cell proportions in the ovaries and spleens of POI model mice. Cell experiments revealed that dhBBR promoted CD4 + cell differentiation into Treg cells. Co-immunoprecipitation revealed Rheb as the dhBBR target that bound to mTOR. However, MHY1485 restored dhBBR-induced changes in forkhead box P3, IL-10, transforming growth factor-β1, IL-17, IL-22, retinoic acid-related orphan receptor-γt and p-mTOR levels in Th17- and Treg-induced CD4 + cells. Conclusion Overall, dhBBR targeted the Rheb/mTOR pathway to promote CD4 + cell differentiation into Treg cells and alleviate POI.

Abstract Background Islet transplantation is a recommended treatment for type 1 diabetes but is limited by donor organ shortage. This study introduces an innovative approach for improving the differentiation and functionality of insulin-producing cells (IPCs) from iPSCs using 3D spheroid formation and hydrogel matrix as an alternative pancreatic islet source. The extracellular matrix (ECM) is crucial for pancreatic islet functionality, but finding the ideal matrix for β-cell differentiation has been challenging. We aimed to advance IPC differentiation and maturation through an esterified collagen hydrogel, comparing its effectiveness with conventional basement membrane extract (BME) hydrogels. Methods iPSCs were differentiated into IPCs using a small molecule-based sequential protocol, followed by spheroid formation in concave microwells. Rheological analysis, scanning electron microscopy, and proteomic profiling were used to characterize the chemical and physical properties of each matrix. IPCs, both in single-cell form and as spheroids, were embedded in either ionized collagen or BME hydrogels, which was followed by assessments of morphological changes, pancreatic islet-related gene expression, insulin secretion, and pathway activation using comprehensive analytical techniques. Results Esterified collagen hydrogels markedly improved the structural integrity, insulin expression, and cell-cell interactions in IPC spheroids, forming densely packed insulin-expressing clusters, in contrast to the dispersed cells observed in BME cultures. Collagen hydrogel significantly enhanced the mRNA expression of crucial endocrine markers and maturation factors, with IPC spheroids showing accelerated differentiation from day 5, suggesting a faster differentiation compared to single cells in hydrogel encapsulation. Insulin secretion in response to glucose in collagen environments, with a GSIS index of 2.46 ± 0.05, exceeded those in 2D and BME, demonstrating superior pancreatic islet functionality. Pathway analysis highlighted enhanced insulin secretion capabilities, evidenced by the upregulation of genes like Secretogranin III and Chromogranin A in collagen cultures. In vivo transplantation results showed that collagen hydrogel enhanced cluster integrity, tissue integration, and insulin secretion compared to non-embedded IPCs and BME groups. Conclusion Esterified collagen hydrogels demonstrated superior efficacy over 2D and BME in promoting IPC differentiation and maturation, possibly through upregulation of the expression of key secretion pathway genes. Our findings suggest that using collagen hydrogels presents a promising approach to enhance insulin secretion efficiency in differentiating pancreatic β-cells, advancing cell therapy in diabetes cell therapy.

Abstract Background Age-related reproductive aging is a natural and irreversible physiological process, and delaying childbearing is increasingly common all over the world. Transplantation of mesenchymal stem cells (MSCs) is considered a new and effective therapy to restore ovarian function, but the relevant mechanisms remain unclear. Recently, it has been found that there is a local Renin–angiotensin system (RAS) in human ovary and it plays a key role. Methods After collecting follicular fluid from women who received oocyte retrieval for pure male factor infertility, the level of RAS components in it were detected, and the correlation analysis by linear regression. Then, the in vivo experiments on female C57BL/6 mice were designed to measure ovarian function, and the transcription and translation levels of RAS pathway were detected by molecular biology methods. Moreover, the role of RAS in regulating inflammation and oxidative stress in the co-culture system were explored in in vitro experiments on KGN cells. Results First, a total of 139 samples of analyzable follicular fluid were obtained. The local RAS of ovary, which is independent of systemic RAS (P > 0.05), is affected by age (Pearson r 0.05). Conclusions Together, our findings indicate that a novel possible mechanism to explain how stem cells restore age-related ovarian functional decline.

Abstract Background Kidney fibrosis is a hallmark of chronic kidney disease (CKD) and compromises the viability of transplanted human bone marrow-derived mesenchymal stromal cells (BM-MSCs). Hence, BM-MSCs were genetically-engineered to express the anti-fibrotic and renoprotective hormone, human relaxin-2 (RLX) and green fluorescent protein (BM-MSCs-eRLX + GFP), which enabled BM-MSCs-eRLX + GFP delivery via a single intravenous injection. Methods BM-MSCs were lentiviral-transduced with human relaxin-2 cDNA and GFP, under a eukaryotic translation elongation factor-1α promoter (BM-MSCs-eRLX + GFP) or GFP alone (BM-MSCs-eGFP). The ability of BM-MSCs-eRLX + GFP to differentiate, proliferate, migrate, produce RLX and cytokines was evaluated in vitro, whilst BM-MSC-eRLX + GFP vs BM-MSCs-eGFP homing to the injured kidney and renoprotective effects were evaluated in preclinical models of ischemia reperfusion injury (IRI) and high salt (HS)-induced hypertensive CKD in vivo. The long-term safety of BM-MSCs-RLX + GFP was also determined 9-months after treatment cessation in vivo. Results When cultured for 3- or 7-days in vitro, 1 × 106 BM-MSCs-eRLX + GFP produced therapeutic RLX levels, and secreted an enhanced but finely-tuned cytokine profile without compromising their proliferation or differentiation capacity compared to naïve BM-MSCs. BM-MSCs-eRLX + GFP were identified in the kidney 2-weeks post-administration and retained the therapeutic effects of RLX in vivo. 1–2 × 106 BM-MSCs-eRLX + GFP attenuated the IRI- or therapeutically abrogated the HS-induced tubular epithelial damage and interstitial fibrosis, and significantly reduced the HS-induced hypertension, glomerulosclerosis and proteinuria. This was to an equivalent extent as RLX and BM-MSCs administered separately but to a broader extent than BM-MSCs-eGFP or the angiotensin-converting enzyme inhibitor, perindopril. Additionally, these renoprotective effects of BM-MSCs-eRLX + GFP were maintained in the presence of perindopril co-treatment, highlighting their suitability as adjunct therapies to ACE inhibition. Importantly, no major long-term adverse effects of BM-MSCs-eRLX + GFP were observed. Conclusions BM-MSCs-eRLX + GFP produced greater renoprotective and therapeutic efficacy over that of BM-MSCs-eGFP or ACE inhibition, and may represent a novel and safe treatment option for acute kidney injury and hypertensive CKD.

Abstract Background Spinal cord injury (SCI) is a severe traumatic spinal condition with a poor prognosis. In this study, a scaffold called linearly ordered collagen aggregates (LOCAS) was created and loaded with induced pluripotent stem cells (iPSCs)-derived neural stem cells (NSCs) from human umbilical cord blood derived mesenchymal stem cells (hUCB-MSCs) to treat SCI in a rat model. Methods The rats underwent a complete transection SCI resulting in a 3-mm break at either the T9 or T10 level of the spinal cord. Results Scanning electron microscope analysis revealed a uniform pore structure on the coronal plane of the scaffold. The LOCAS had a porosity of 88.52% and a water absorption of 1161.67%. Its compressive modulus and stress were measured at 4.1 MPa and 205 kPa, respectively, with a degradation time of 10 weeks. After 12 weeks, rats in the LOCAS-iPSCs-NSCs group exhibited significantly higher BBB scores (8.6) compared to the LOCAS-iPSCs-NSCs group (5.6) and the Model group (4.2). The CatWalk analysis showed improved motion trajectory, regularity index (RI), and swing speed in the LOCAS-iPSCs-NSCs group compared to the other groups. Motor evoked potentials latency was lower and amplitude was higher in the LOCAS-iPSCs-NSCs group, indicating better neural function recovery. Histological analysis demonstrated enhanced neuronal differentiation of NSCs and nerve fiber regeneration promoted by LOCAS-iPSCs-NSCs, leading to improved motor function recovery in rats. The LOCAS scaffold facilitated ordered neurofilament extension and guided nerve regeneration. Conclusions The combination of LOCAS and iPSCs-NSCs demonstrated a positive therapeutic impact on motor function recovery and tissue repair in rats with SCI. This development offers a more resilient bionic microenvironment and presents novel possibilities for clinical SCI repair.

Abstract The human endometrium is a highly regenerative tissue capable of undergoing scarless repair during the menstruation and postpartum phases. This process is mediated by endometrial adult stem/progenitor cells. During the healing of endometrial injuries, swift reepithelization results in the rapid covering of the wound surface and facilitates subsequent endometrial restoration. The involvement of endogenous endometrial epithelial stem cells, stromal cells, and bone marrow-derived cells in the regeneration of the endometrial epithelium has been a subject of prolonged debate. Increasing evidence suggests that the regeneration of the endometrial epithelium mainly relies on epithelial stem cells rather than stromal cells and bone marrow-derived cells. Currently, no consensus has been established on the identity of epithelial stem cells in the epithelial compartment. Several markers, including stage-specific embryonic antigen-1 (SSEA-1), sex-determining region Y-box 9 (SOX9), neural-cadherin (N-cadherin), leucine-rich-repeat-containing G-protein-coupled receptor 5 (LGR5), CD44, axis inhibition protein 2 (Axin2), and aldehyde dehydrogenase 1A1 (ALDH1A1), have been suggested as potential candidate markers for endometrial epithelial stem cells. The identification of endometrial epithelial stem cells contributes to our understanding of endometrial regeneration and offers new therapeutic insights into diseases characterized by regenerative defects in the endometrium, such as intrauterine adhesion. This review explores different perspectives on the origins of human and mouse endometrial epithelial cells. It summarizes the potential markers, locations, and hierarchies of epithelial stem cells in both human and mouse endometrium. It also discusses epithelial cell-based treatments for intrauterine adhesion, hoping to inspire further research and clinical application of endometrial epithelial stem cells. Graphical abstract

Abstract Backgroud Intrauterine Adhesions (IUA) is a common gynecological disease which is seriously endangers the reproductive function of women without any ideal treatment. Some researchers found Menstrual Blood-derived Mesenchymal Stem Cells (MenSCs) can repair of damaged endometrium, however, has not been fully clarified. This study aims to evaluate the therapeutic effects of MenSCs in IUA and the repair mechanism in vivo. Methods This study is Laboratory-based study. To evaluate the therapeutic effects of MenSCs in IUA, We cultivated MenSCs, established mouse endometrial injury model, observed the uterine morphology and degree of endometrial fibrosis and compared the expression of CXC chemokine ligand-13 (CXCL13)、CXC chemokine receptor-5 (CXCR5)、Plasmin Activating Inhibitor-1(Pai-1), Transforming Growth Faction-β1(TGF- β1) and Matrix Metalloproteinase-9 (Mmp-9) among each groups. GraphPad Prism 8.0 was used for statistical processing. Data were expressed as mean ± SD. Statistical comparisons among groups were performed with one-way ANOVA. P

Abstract Background Liver fibrosis is a common pathological process of chronic liver disease, characterized by excessive deposition of extracellular matrix (ECM). Mesenchymal stem cells (MSCs) have been found to have potential therapy effect on liver fibrosis, but the mechanism involved was still unclear. The objective of this study is to investigate the therapeutic efficacy of adipose-derived mesenchymal stem cells (ADMSCs) on the treatment of liver fibrosis, with particular emphasis on elucidating the underlying mechanism of action through which ADMSCs inhibit the activation of hepatic stellate cells (HSCs). Methods ADMSCs were isolated from adipose tissue and injected intravenously into hepatic fibrosis model of rats. The histopathological changes, liver function, collagen deposition, the expression of fibroin and Hippo pathway were evaluated. In vitro, ADMSCs were co-cultured with HSCs activated by transforming growth factor beta 1 (TGF-β1), and the inhibitor of Hippo pathway was used to evaluate the therapeutic mechanism of ADMSCs transplantation. Results The results showed that after the transplantation of ADMSCs, the liver function of rats was improved, the degree of liver fibrosis and collagen deposition were reduced, and the Hippo signaling pathway was activated. In vitro, ADMSCs can effectively inhibit the proliferation and activation of HSCs induced by TGF-β1 treatment. However, the inhibitory effect of ADMSCs was weakened after blocking the Hippo signaling pathway. Conclusions ADMSCs inhibit HSCs activation by regulating YAP/TAZ, thereby promoting functional recovery after liver fibrosis. These findings lay a foundation for further investigation into the precise mechanism by which ADMSCs alleviate liver fibrosis.

Abstract Background Radiation-induced skin injury (RISI) represents a significant complication in patients receiving radiotherapy and individuals exposed to nuclear accidents, characterized by a protracted wound-healing process relative to injuries from other etiologies. Current preventive and management approaches remain inadequate. Consequently, investigating efficacious intervention strategies that target the disease’s progression characteristics holds significant practical importance. Methods Small interfering RNA (siRNA) and overexpression plasmid were used to modulate the expression of Marvel domain containing 3 (Marveld3) and paired related homeobox 2 (PRRX2). Protein and mRNA levels were estimated by Western Blot and real-time PCR, respectively. Intracellular levels of Malondialdehyde (MDA), a terminal product of lipid peroxidation, were measured following the manufacturer’s protocol for MDA assay kit. Similarly, intracellular levels of ferrous iron (Fe2+) and reactive oxygen species (ROS) were determined using their respective assay kits. Lipid peroxidation status within the cells was evaluated via BODIPY staining. Immunohistochemistry was conducted to ascertain the expression of PRRX2 in skin tissues collected at various time points following irradiation of rats. The H-score method was used to evaluate the percentage of positively stained cells and staining intensity. RNA sequencing, Gene Ontology (GO) analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were conducted by OE Biotech Company. Results In this study, our findings indicated that Marveld3 suppression could effectively inhibit lipid peroxidation levels in irradiated skin cells, concomitantly reducing intracellular Fe2+ content. Additionally, the silencing of Marveld3 effectively abrogated the impact of a ferroptosis agonist on cellular viability, resulting in the upregulation of 66 and 178 genes, as well as the downregulation of 188 and 31 genes in irradiated HaCaT and WS1 cells, respectively. Among the differentially expressed genes, the PRRX2 which was found to be involved in the process of ferroptosis, exhibited statistically significant upregulation. And the upregulation of PRRX2 expression may attenuate radiation-induced lipid peroxidation in skin cells, thereby functioning as a potential stress-responsive mechanism to counteract radiation effects. Conclusions This study elucidates the role of Marveld3 in radiation-induced ferroptosis in skin cells. Inhibition of Marveld3 led to the upregulation of PRRX2, which subsequently resulted in a reduction of Fe2+ and ROS levels, as well as the suppression of lipid peroxidation. These effects collectively mitigated the occurrence of ferroptosis.

Abstract Background Sepsis-associated acute kidney injury (AKI) is a serious complication of systemic infection with high morbidity and mortality in patients. However, no effective drugs are available for AKI treatment. Dexmedetomidine (DEX) is an alpha 2 adrenal receptor agonist with antioxidant and anti-apoptotic effects. This study aimed to investigate the therapeutic effects of DEX on sepsis-associated AKI and to elucidate the role of mitochondrial dynamics during this process. Methods A lipopolysaccharide (LPS)-induced AKI rat model and an NRK-52E cell model were used in the study. This study investigated the effects of DEX on sepsis-associated AKI and the molecular mechanisms using histologic assessment, biochemical analyses, ultrastructural observation, western blotting, immunofluorescence, immunohistochemistry, qRT-PCR, flow cytometry, and si-mRNA transfection. Results In rats, the results showed that administration of DEX protected kidney structure and function from LPS-induced septic AKI. In addition, we found that DEX upregulated the α2-AR/SIRT1/PGC-1α pathway, protected mitochondrial structure and function, and decreased oxidative stress and apoptosis compared to the LPS group. In NRK-52E cells, DEX regulated the mitochondrial dynamic balance by preventing intracellular Ca2+ overloading and activating CaMKII. Conclusions DEX ameliorated septic AKI by reducing oxidative stress and apoptosis in addition to modulating mitochondrial dynamics via upregulation of the α2-AR/SIRT1/PGC-1α pathway. This is a confirmatory study about DEX pre-treatment to ameliorate septic AKI. Our research reveals a novel mechanistic molecular pathway by which DEX provides nephroprotection.

Abstract Background Overconsumption of retinoic acid (RA) or its analogues/derivatives has been linked to severe craniomaxillofacial malformations, such as cleft palate and midface hypoplasia. It has been noted that RA disturbed the proliferation and migration of embryonic palatal mesenchymal (EPM) cells in these malformations, yet the exact mechanisms underlying these disruptions remained unclear. Methods A model of retinoic acid (RA)-induced cleft palate in fetal mice was successfully established. Histological alterations in the palate were evaluated using Hematoxylin and Eosin (H&E) staining and RNA in situ hybridization (RNAscope). Cellular proliferation levels were quantified via the Cell Counting Kit-8 (CCK-8) assay and EdU incorporation assay, while cell migration capabilities were investigated using wound healing and Transwell assays. Mitochondrial functions were assessed through Mito-Tracker fluorescence, mitochondrial reactive oxygen species (ROS) measurement, ATP level quantification, and mitochondrial DNA (mtDNA) copy number analysis. Differential gene expression and associated signaling pathways were identified through bioinformatics analysis. Alterations in the transcriptional and translational levels of Lhx6 and genes associated with mitophagy were quantified using quantitative PCR (qPCR) and Western blot analysis, respectively. Mitochondrial morphology and the mitochondrial autophagosomes within cells were examined through transmission electron microscopy (TEM). Results Abnormal palatal development in mice, along with impaired proliferation and migration of human embryonic palatal mesenchymal (HEPM) cells, was associated with RA affecting mitochondrial function and concomitant downregulation of Lhx6. Knockdown of Lhx6 in HEPM cells resulted in altered cell proliferation, migration, and mitochondrial function. Conversely, the aberrant mitochondrial function, proliferation, and migration observed in RA-induced HEPM cells were ameliorated by overexpression of Lhx6. Subsequent research demonstrated that Lhx6 ameliorated RA-induced dysfunction in HEPM cells by modulating PINK1/Parkin-mediated mitophagy, thereby activating the MAPK signaling pathways. Conclusion Lhx6 is essential for mitochondrial homeostasis via tuning PINK1/Parkin-mediated mitophagy and MAPK signaling pathways. Downregulation of Lhx6 by RA transcriptionally disturbs the mitochondrial homeostasis, which in turn leads to the proliferation and migration defect in HEPM cells, ultimately causing the cleft palate. Graphical abstract

Abstract Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the loss of motoneurons (MNs), and despite progress, there is no effective treatment. A large body of evidence shows that astrocytes expressing ALS-linked mutant proteins cause non-cell autonomous toxicity of MNs. Although MNs innervate muscle fibers and ALS is characterized by the early disruption of the neuromuscular junction (NMJ) and axon degeneration, there are controversies about whether muscle contributes to non-cell-autonomous toxicity to MNs. In this study, we generated primary skeletal myotubes from myoblasts derived from ALS mice expressing human mutant SOD1G93A (termed hereafter mutSOD1). Characterization revealed that mutSOD1 skeletal myotubes display intrinsic phenotypic and functional differences compared to control myotubes generated from non-transgenic (NTg) littermates. Next, we analyzed whether ALS myotubes exert non-cell-autonomous toxicity to MNs. We report that conditioned media from mutSOD1 myotubes (mutSOD1-MCM), but not from control myotubes (NTg-MCM), induced robust death of primary MNs in mixed spinal cord cultures and compartmentalized microfluidic chambers. Our study further revealed that applying mutSOD1-MCM to the MN axonal side in microfluidic devices rapidly reduces mitochondrial axonal transport while increasing Ca2 + transients and reactive oxygen species (i.e., H2O2). These results indicate that soluble factor(s) released by mutSOD1 myotubes cause MN axonopathy that leads to lethal pathogenic changes.

Abstract Background CHRDL1 belongs to a novel class of mRNA molecules. Nonetheless, the specific biological functions and underlying mechanisms of CHRDL1 in oral squamous cell carcinoma (OSCC) remain largely unexplored. Methods RT-qPCR and immunohistochemical staining were employed to assess the mRNA and protein expression levels of the MED29 gene in clinical samples of OSCC. Additionally, RT-qPCR and Western Blot analyses were conducted to investigate the mRNA and protein expression levels of the MED29 gene specifically in OSCC. The impact of MED29 on epithelial-mesenchymal transition (EMT), invasion, and migration of OSCC was evaluated through scratch assay, transwell assay, and immunofluorescence staining. Furthermore, wound healing assay and Transwell assay were utilized to examine whether CHRDL1 influences the malignant behavior of OSCC by modulating MED29 in vitro. The regulatory role of CHRDL1 on MED29 was further elucidated in vivo through a tail vein lung metastasis model in nude mice. Results MED29 expression was elevated in tumor tissues of OSCC patients compared with adjacent cancer tissues. Moreover, in CAL27 and SCC25 cell lines, MED29 was upregulated and associated with increased cell migration and invasion abilities. Overexpression of MED29 facilitated EMT in OSCC cell lines, whereas knockdown of MED29 impeded EMT, resulting in diminished cell migration and invasion capacities. CHRDL1 exerted inhibitory effects on the expression of MED29, thereby suppressing EMT progression and consequently restraining the invasion and migration of OSCC cells. Furthermore, CHRDL1 mediated the inhibition of migration of OSCC cell lines to the OSCC through its regulation of MED29. Conclusions MED29 facilitated the epithelial-mesenchymal transition process in OSCC, thereby promoting migration and invasion. On the other hand, CHRDL1 exerted inhibitory effects on the invasion and metastasis of OSCC by suppressing MED29 through the inhibition of the MAPK signaling pathway.