An integrative pharmacology-based study on the efficacy and mechanism of essential oil of Chaihu Guizhi Decoction on influenza A virus induced pneumonia in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Chaihu Guizhi Decoction (CGD) has a long history of use in China for the treatment of influenza, which involves the use of a variety of aromatic herbs. Our previous studies have found that the contents of aromatic constituents in CGD affected the efficacy of treatment of influenza-infected mice, suggesting a clue that essential oil from CGD may play a relatively important role in ameliorating influenza induced pneumonia. AIM OF THE STUDY: To evaluate the anti-influenza potential of essential oil derived from Chaihu Guizhi Decoction (CGD-EO), to characterize and predict the key active components in CGD-EO, and to explore the mechanism of action of CGD-EO. MATERIALS AND METHODS: CGD-EO was obtained by steam distillation, and the components of the essential oil were characterized by gas chromatography-mass spectrometry (GC-MS) in conjunction with the retention index. The constituents absorbed into the blood of mice treated with CGD-EO were analyzed by headspace solid phase microextraction gas chromatography/mass spectrometry (HS-SPME-GC/MS). The potential anti-influenza active constituents and their possible action pathway were predicted by simulation using a network pharmacology approach. The protective effect of CGD-EO and its major components on H1N1/PR8-infected cells was determined using the CCK8 assay kit. Mice infected with influenza A virus H1N1/PR8 were administered different doses of CGD-EO orally and the body weights and lung weights were recorded. Mice with varying degrees of H1N1/PR8 infection were administered CGD-EO orally, and their daily weight, water consumption, and clinical indicators were recorded. Necropsies were conducted on days 3 and 5, during which lung weights were measured and lung tissues were preserved. Furthermore, the mRNA expression of the H1N1/PR8 virus and inflammatory factors in lung tissue was analyzed using RT-qPCR. RESULTS: (E)-cinnamaldehyde was the most abundant compound in the CGD-EO. The results of serum medicinal chemistry combined with network pharmacological analysis indicated that (E)-cinnamaldehyde and 3-phenyl-2-propenal may be potential active components of the CGD-EO anti-influenza, and may be involved in the NF-κB signalling pathway. In vitro studies have demonstrated that both CGD-EO and cinnamaldehyde exert a protective effect on MDCK cells infected with H1N1/PR8. In a 0.5 TCID50 H1N1/PR8-induced influenza model, mice treated with CGD-EO at a dose of 63.50 µg/kg exhibited a reduction in lung index, pathological lung lesions, and H1N1/PR8 viral gene levels. In addition, CGD-EO treatment was found to regulate the levels of inflammatory cytokines, including IL-6, TNF-α, and IFN-γ. Moreover, following three days of administration, an upregulation of NF-κB mRNA levels in mouse lung tissue was observed in response to CGD-EO treatment. CONCLUSIONS: The findings of our study indicate CGD-EO exerts a protective effect against H1N1-induced cytopathic lesions in vitro and is capable of alleviating H1N1-induced pneumonitis in mice. Moreover, it appears to be more efficacious in the treatment of mild symptoms of H1N1 infection. Studies have demonstrated that CGD-EO has antiviral potential to attenuate influenza-induced lung injury by modulating inflammatory cytokines and NF-κB signalling pathways during the early stages of influenza infection. It is possible that (E)-cinnamaldehyde is a potential active ingredient in the anti-influenza efficacy of CGD-EO.

When the Bough Breaks: The Financial Burden of Childbirth and Postpartum Care by Insurance Type.

Policy Points This study examines exposure to out-of-pocket (OOP) costs related to childbirth and postpartum care for those with a Medicaid-insured birth compared with those with a commercially insured birth and subsequent financial outcomes at 12 months postpartum. We find that Medicaid is highly protective against health care costs for childbirth and postpartum care relative to commercial insurance, particularly for birthing people with low income. We find persistent medical debt and worry at 12 months postpartum for Medicaid recipients who reported OOP childbirth expenses. CONTEXT: Out-of-pocket (OOP) costs related to childbirth and postpartum care may cause financial hardship, depending on type of insurance and income. METHODS: We estimated OOP spending on childbirth and postpartum care and financial strain 1 year after birth, comparing Medicaid-insured births with commercially insured births. The Postpartum Assessment of Health Survey followed up with respondents to the Centers for Disease Control and Prevention (CDC) Pregnancy Risk Assessment Monitoring System after a 2020 birth in six states and New York City. The survey included questions on health care costs and financial well-being. Our analytic sample consisted of 4,453 postpartum people, 1,544 with a Medicaid-insured birth and 2,909 with a commercially insured birth. FINDINGS: We observe significant financial hardship from childbirth that persists into the postpartum year, with significant differences by insurance and income. We find Medicaid is highly financially protective relative to commercial insurance; 81.4% of Medicaid-insured births were free to the patient, compared with 15.7% of commercially insured births (p < 0.001). Six of ten commercially insured births (59%) cost over $1,000 OOP. Among respondents reporting OOP costs for childbirth, we found that Medicaid enrollees are more likely to have borrowed money from friends or family to pay for childbirth (8% vs. 1%, p < 0.001) and one in five had not made any payments 1 year postpartum (26% vs. 5% of commercially insured births, p < 0.001). Among the commercially insured, those with incomes under 200% of the federal poverty level (FPL) fared worse financially than those above 200% FPL on a number of indicators, including debt in collection (33% vs. 13%, p < 0.001) and financial worry (55% vs. 34%, p < 0.001). CONCLUSIONS: The cost of childbirth and postpartum health care results in significant and persistent financial hardship, particularly for families with lower income with commercial insurance. Medicaid offers greater protection for families with low income by offering reduced cost sharing for childbirth and postpartum health care, but even minimal cost sharing in Medicaid causes financial strain.

Spermine Synthase : A Potential Prognostic Marker for Lower-Grade Gliomas.

Objective: The objective of this study was to assess the relationship between spermine synthase (SMS) expression, tumor occurrence, and prognosis in lower-grade gliomas (LGGs). Methods: A total of 523 LGG patients and 1152 normal brain tissues were included as controls. Mann-Whitney U test was performed to evaluate SMS expression in the LGG group. Functional annotation analysis was conducted to explore the biological processes associated with high SMS expression. Immune cell infiltration analysis was performed to examine the correlation between SMS expression and immune cell types. The association between SMS expression and clinical and pathological features was assessed using Spearman correlation analysis. In vitro experiments were conducted to investigate the effects of overexpressing or downregulating SMS on cell proliferation, apoptosis, migration, invasion, and key proteins in the protein kinase B (AKT)/epithelialmesenchymal transition signaling pathway. Results: The study revealed a significant upregulation of SMS expression in LGGs compared to normal brain tissues. High SMS expression was associated with certain clinical and pathological features, including older age, astrocytoma, higher World Health Organization grade, poor disease-specific survival, disease progression, non-1p/19q codeletion, and wild-type isocitrate dehydrogenase. Cox regression analysis identified SMS as a risk factor for overall survival. Bioinformatics analysis showed enrichment of eosinophils, T cells, and macrophages in LGG samples, while proportions of dendritic (DC) cells, plasmacytoid DC (pDC) cells, and CD8+ T cells were decreased. Conclusion: High SMS expression in LGGs may promote tumor occurrence through cellular proliferation and modulation of immune cell infiltration. These findings suggest the prognostic value of SMS in predicting clinical outcomes for LGG patients.

Methionine restriction promotes cisplatin sensitivity of gastric cancer resistant cells by down-regulating circ-CDK13 level.

BACKGROUND: Methionine restriction (MR) is a research direction in the treatment of gastric cancer (GC). The aim of this study was to investigate the molecular mechanism of MR on enhancing cisplatin (DDP) sensitivity of drug-resistant GC cells. METHODS: Twenty pairs of GC tissues and adjacent normal gastric mucosa tissues were collected. DDP-resistant cell lines (KATO/DDP and MKN45/DDP), mouse model of GC and GC patient-derived organoid (PDO) models were established. Lentivirus-mediated METase overexpression was used for MR. Cell viability and apoptosis were detected by MTT assay and flow cytometry. Western blotting was used to detect multi-drug resistance-1 (MDR1), MDR-associated protein 1 (MRP1) eukaryotic initiation factor 4A-Ⅲ (EIF4A3), and METase protein expressions. The levels of circRNAs were detected by qRT-PCR. Tumor volume and weight were measured. The proliferation of tumor cells was detected by immunohistochemical staining. RESULTS: The differentially expressed circRNAs of GC were screened in Gene Expression Omnibus database. MR in KATO/DDP and MKN45/DDP cells significantly down-regulated circ-CDK13 level. Overexpression of circ-CDK13 significantly inhibited apoptosis of sensitive cells (KATO III and MKN45). Interference with circ-CDK13 significantly promoted apoptosis of drug-resistant cells (KATO/DDP and MKN45/DDP). MR enhanced the DDP sensitivity of GC resistant cells, GC PDO and GC mice by down-regulating circ-CDK13. EIF4A3 binds to the downstream flanking sequence of circ-CDK13, and interference with EIF4A3 reduces circ-CDK13 levels, but does not affect CDK13. The expressions of circ-CDK13 and EIF4A3 in GC clinical samples were increased and positively correlated. Simultaneously overexpression of METase and EIF4A3 in resistant cells inhibited apoptosis, and further interference with circ-CDK13 reversed this effect. CONCLUSION: MR inhibits circ-CDK13 level by down-regulating EIF4A3, thereby increasing the sensitivity of GC drug-resistant cells to DDP.

Addition of neutrophil-to-lymphocyte ratio to Pre-DAA FIB-4 does not increase prediction value for de novo liver complications in hepatitis C.

BACKGROUND AND AIMS: Direct-acting antiviral agents (DAAs) achieve high sustained virologic response (SVR) in chronic hepatitis C patients; yet a proportion of patients still experience de novo liver complications after SVR. Identification of risk factors is clinically important. FIB-4 index is a useful noninvasive tool to assess fibrosis, while neutrophil-to-lymphocyte ratio (NLR) is a biomarker for systemic inflammation. Our study aimed to investigate whether the addition of NLR can increase the prediction power of pre-DAA FIB-4 for de novo liver complications after SVR. METHODS: We recruited patients via The Taiwan HCV Registry (TACR) and National Health Insurance Registry Database. The inclusion criteria were patients who achieved SVR12 after DAA and were followed for at least 24 months after SVR12. Liver complications included ascites, hepatic encephalopathy, variceal bleeding, and HCC. RESULTS: Totally 7657 patients were recruited from 2013 to 2018. Among them, 3674 patients (48.0%) had a FIB-4 value > 3.25 and 491 patients (6.4%) had a NLR >4 before DAA. After two-year of follow-up after SVR 12, 214 patients (2.8%) developed de novo liver complications. Factors associated with liver complications included male gender, diabetes mellitus, hyperlipidemia, chronic kidney disease, and pre-DAA FIB-4 >3.25 in multivariate analyses. Addition of NLR slightly did not increase the power of predicting liver complications. CONCLUSIONS: The overall incidence of de novo liver complications after SVR is low during short-term follow-up. Elevated pre-DAA FIB-4 is associated with de novo liver complications after SVR, whereas the addition of pre-DAA NLR does not increase the prediction power.

A review of chyme reinfusion: new tech solutions for age old problems.

High-output double enterostomies (DES) and enteroatmospheric fistulae (EAF) are associated with significant morbidity, including infection, malnutrition, and prolonged hospital admissions. Management is complex and has remained a challenging surgical problem for many decades in both adult and paediatric patient populations. Chyme reinfusion (CR) from the proximal to distal DES or EAF limb is a potential therapeutic solution which has been shown to be safe and beneficial; however, early methods have involved the manual handling of chyme, which is labour intensive and poorly tolerated by both patients and staff. Over the past four decades, there has been growing interest in the application and development of medical device technology to improve the effectiveness and user-friendliness of CR. New Zealand (NZ) has been at the forefront of innovation in this field, with exciting translational research projects in both adults and neonates (funded and enabled by the NZ MedTech CORE). This narrative review provides a summary of the evolution of CR technology globally, synthesises the extant clinical evidence and highlights future directions.

Porphyromonas gingivalis GroEL accelerates abdominal aortic aneurysm formation by matrix metalloproteinase-2 SUMOylation in vascular smooth muscle cells: A novel finding for the activation of MMP-2.

Infection is a known cause of abdominal aortic aneurysm (AAA), and matrix metalloproteases-2 (MMP-2) secreted by vascular smooth muscle cells (SMCs) plays a key role in the structural disruption of the middle layer of the arteries during AAA progression. The periodontal pathogen Porphyromonas gingivalis is highly associated with the progression of periodontitis. GroEL protein of periodontal pathogens is an important virulence factor that can invade the body through either the bloodstream or digestive tract and is associated with numerous systemic diseases. Although P. gingivalis aggravates AAA by increasing the expression of MMP-2 in animal studies, the molecular mechanism through which P. gingivalis regulates the expression of MMP-2 is still unknown and requires further investigation. In this study, we first confirmed through animal experiments that P. gingivalis GroEL promotes MMP-2 secretion from vascular SMCs, thereby aggravating Ang II-induced aortic remodeling and AAA formation. In addition, rat vascular SMCs and A7r5 cells were used to investigate the underlying mechanisms in vitro. The results demonstrated that GroEL can promote the interaction between the K639 site of MMP-2 and SUMO-1, leading to MMP-2 SUMOylation, which inhibits the reoccurrence of non-K639-mediated monoubiquitylation. Hence, the monoubiquitylation-mediated lysosomal degradation of MMP-2 is inhibited, consequently promoting MMP-2 stability and production. SUMOylation may facilitate intra-endoplasmic reticulum (ER) and Golgi trafficking of MMP-2, thereby enhancing its transport capacity. In conclusion, this is the first report demonstrating the presence of a novel posttranslational modification, SUMOylation, in the MMP family, suggesting that P. gingivalis GroEL may exacerbate AAA formation by increasing MMP-2 production through SUMOylation in vascular SMCs. This study also provides a novel perspective on the role of SUMOylation in MMP-2-induced systemic diseases.

Chemoselective Pd-Based Dyotropic Rearrangement: Fluorocyclization and Regioselective Wacker Reaction of Homoallylic Amides.

Fluorocyclization of alkenes tethered with a pronucleophile is an efficient transformation that converts easily accessible starting materials to fluorinated heterocycles in a single step. We report herein an unprecedented Pd(II)-catalyzed oxidative domino process that transforms homoallylic amides to 5,6-dihydro-4H-1,3-oxazines through a domino oxypalladation/PdII-oxidation/dyotropic rearrangement/reductive elimination sequence. Three chemical bonds are created under these operationally simple conditions. Taking advantage of the facile hydrolysis of the α-fluoro tertiary alkyl ether under acidic conditions, a one-pot conversion of homoallylic amides to homologated ketones is subsequently developed, which represents a rare example of regioselective Wacker oxidation reaction of 1,1-disubstituted alkenes.

Exploring the changing landscape of medical imaging: insights from highly cited studies before and during the COVID-19 pandemic.

OBJECTIVES: To investigate whether and how the COVID-19 pandemic has changed medical imaging trends by synthesizing the highly cited studies before and during the pandemic. METHODS: In this cross-sectional study, we identified highly cited studies on medical imaging from the essential science indicators (ESI) database, categorizing them into two periods: before the pandemic (January 2016-December 2019) and during the pandemic (January 2020-December 2023). We conducted a global research landscape comparative analysis and utilized CiteSpace and VOSviewer software to create knowledge maps for analyzing the co-occurrences of keywords and references in this field. RESULTS: A total of 2914 highly cited studies were included in this study, which revealed a notable 30.1% increase in medical imaging publications during the pandemic. Enhanced international cooperation has been observed, with European countries and the US leading the research efforts. Keyword analysis revealed that artificial intelligence (AI) has remained a dominant hotspot in medical imaging research before and during the pandemic. References analysis showed a shift in focus towards COVID-19-related studies, overshadowing some important areas including cancer imaging, cardiac imaging, and neuroimaging. CONCLUSIONS: Over the past four years, the COVID-19 pandemic has led to changes in the research output, international collaborations, and hotspots within highly cited medical imaging studies. Navigating the post-COVID era, it is imperative to continue fostering international collaboration, prioritize resource allocation to refocus on overlooked research areas, and develop long-term strategic plans to prepare for and mitigate the impact of future public health crises. KEY POINTS: Question Understanding how the COVID-19 pandemic has changed medical imaging trends and priorities, which is crucial for preparing against future public health crises, remains unclear. Findings The COVID-19 pandemic has led to increases in highly cited medical imaging studies, enhancements in international collaborations, and shifts in research hotspots. Clinical relevance This study provides a comprehensive overview of highly cited studies on medical imaging before and during the pandemic, highlighting the pivotal role of AI in the post-COVID era and emphasizing the need to refocus on potentially neglected research areas.

Recent breakthroughs in innovative elements, multidimensional enhancements, derived technologies, and novel applications of PROTACs.

Proteolysis Targeting Chimera (PROTAC) is an emerging and evolving technology based on targeted protein degradation (TPD). Small molecule PROTACs have shown great efficacy in degrading disease-specific proteins in preclinical and clinical studies, but also showed various limitations. In recent years, new technologies and advances in TPD have provided additional optimized strategies based on conventional PROTACs that can overcome the shortcomings of conventional PROTACs in terms of undruggable targets, bioavailability, tissue-specificity, spatiotemporal control, and degradation scope. In addition, some designs of special targeting chimeras and applications based on multidisciplinary science have shed light on novel therapeutic modalities and drug design. However, each improvement has its own advantages, disadvantages and application conditions. In this review, we summarize the exploration of PROTAC elements, depict a landscape of improvements and derived concepts of PROTACs, and expect to provide perspectives for technological innovations, combinations and applications in future targeting chimera design.

The role of AKR1B10 in osteogenic differentiation of mesenchymal stem cells and atrophic nonunion.

Atrophic nonunion is a chronic disease without effective medications. Here, high-throughput mRNA sequencing was used to explore the novel targets in atrophic nonunion. AKR1B10, a member of aldo-keto reductase family 1, is upregulated in atrophic nonunion tissues. There are currently no studies to reveal the role of AKR1B10 in atrophic nonunion. We used rat bone marrow-derived mesenchymal stem cells (BMSCs) to explore the effect of AKR1B10 on the osteogenic differentiation and autophagy. In vivo, we implanted collagen sponges loaded with LV-shAKR1B10-transduced BMSCs into rat fractured femurs to explore the role of AKR1B10 in fracture healing. The results showed that AKR1B10 reduced the activity of ALP, suppressed the expression of COL1A1, RUNX2 and OCN, and inhibited calcification deposition in osteogenically differentiated BMSCs. AKR1B10 reduced the expression of LC3II, decreased the number of autophagosomes, and promoted the expression of p62. In addition, the promoting effect of AKR1B10 knockdown on osteogenic differentiation of BMSCs was attenuated by 3-MA treatment. Implantation of collagen sponges found that knockdown of AKR1B10 promoted bone fracture healing. In conclusion, AKR1B10 inhibited the osteogenic differentiation and autophagy, and delayed the bone fracture healing. These results provide a new perspective on revealing the role of AKR1B10 in nonunion and may also provide a new therapeutic target for the treatment of nonunion.

Finite Element Simulation of Residual Stress in Laser Additive Manufactured Functionally Gradient Materials Based on Bessel Heat Source Model.

Laser additive manufacturing (LAM) technology has the advantages of short manufacturing cycles, low material waste rate, and design ability. It is especially suitable for preparing functionally gradient materials (FGM). However, due to the large temperature gradient and the change in material composition, the residual stress is very high, which will seriously affect the mechanical properties and manufacturing accuracy of the structure. In this study, the thermomechanical coupled finite element model based on the Bessel heat source is established, and the residual stress in LAM TC4/TC11 FGM is obtained. The results show that the Bessel heat source can effectively suppress the generation of residual stress in the additive manufacturing process, and the finite element results are consistent with the experimental results. Compared with the traditional Gaussian heat source, the maximal residual tensile stress is reduced by an average of 28.1%. The value of residual stress increases with the increase in the number of printing layers, and it increases with the increase of the laser power and decreases with the increase of the scanning speed. The overall trend is that the two sides are compressive stress and the middle is tensile stress. The research has important reference significance for the reasonable suppression of the residual stress in FGM produced by LAM.

Bibliometric analysis of the inflammation expression after spinal cord injury: current research status and emerging frontiers.

STUDY DESIGN: Bibliometric analysis. OBJECTIVE: To analyze literature on inflammatory expression following spinal cord injury, highlighting development trends, current research status, and potential emerging frontiers. SETTING: Not applicable. METHODS: Articles were retrieved using terms related to spinal cord injury and inflammatory responses from the Web of Science Core Collection, covering January 1, 1980, to May 23, 2024. Tools like CiteSpace and VOSviewer assessed the research landscape, evaluating core authors, journals, and contributing countries. Keyword co-occurrence analyses identified research trends. RESULTS: A total of 2504 articles were retrieved, showing a consistent increase in publications. The Journal of Neurotrauma had the highest publication volume and influence. The most prolific author was Cuzzocrea S, with Popovich PG having the highest H-index. China led in the number of publications, followed closely by the United States, which had the highest impact and extensive international collaboration. Research mainly focused on nerve function recovery, glial scar formation, and oxidative stress. Future research is expected to investigate cellular autophagy, vesicular transport, and related signaling pathways. CONCLUSION: The growing interest in inflammation caused by spinal cord injury is evident, with current research focusing on oxidative stress, glial scar, and neurological recovery. Future directions include exploring autophagy and extracellular vesicles for new therapies. Interdisciplinary research and extensive clinical trials are essential for validating new treatments. Biomarker discovery is crucial for diagnosis and monitoring, while understanding autophagy and signaling pathways is vital for drug development. Global cooperation is needed to accelerate the application of scientific findings, improving spinal cord injury treatment.

Natural synergy: Oleanolic acid-curcumin co-assembled nanoparticles combat osteoarthritis.

Curcumin (Cur) is a natural polyphenol that is one of the most valuable natural products. However, its use as a functional food is limited by low water solubility, chemical instability and poor bioavailability. In this study, a supramolecular co-assembly strategy was used to construct an oleanolic acid-curcumin (OLA-Cur) co-assembly composite nano-slow-release treatment system. As a co-assembled compound, OLA is a widely present pentacyclic triterpenoid compound with multiple biological activities in the plant kingdom, which is expected to jointly alleviate the damaging effects of papain-induced mouse osteoarthritis model. The OLA-Cur NPs shows the solid core-shell structure, which can effectively improve the water solubility of Cur and OLA, and has good stability and sustained release characteristics. The analysis results show that the two compounds are mainly assembled through hydrogen bonding interactions, hydrophobic interactions, and π - π stacking interactions. The OLA-Cur NPs can inhibit the release of pro-inflammatory cytokines TNF-α, IL-6, and IL-1ß induced by LPS in RAW264.7 mouse macrophages, promote the secretion of anti-inflammatory cytokine IL-10, and improve the oxidative stress index of hydrogen peroxide induced human rheumatoid arthritis synovial fibroblasts. In addition, it has a certain improvement effect on cartilage and subchondral bone damage in mouse osteoarthritis models. These findings suggest that constructing co-assembled composite nanoparticles based on pure natural compounds may break through the limitations of a variety of important nutritional ingredients in functional foods.

Use of physiologically based kinetic modeling to predict neurotoxicity and genotoxicity of methylglyoxal in humans.

This study aimed to evaluate human neurotoxicity and genotoxicity risks from dietary and endogenous methylglyoxal (MGO), utilizing physiologically based kinetic (PBK) modeling-facilitated reverse dosimetry as a new approach methodology (NAM) to extrapolate in vitro toxicity data to in vivo dose-response predictions. A human PBK model was defined based on a newly developed and evaluated mouse model enabling the translation of in vitro toxicity data for MGO from human stem cell-derived neurons and WM-266-4 melanoma cells into quantitative human in vivo toxicity data and subsequent risk assessment by the margin of exposure (MOE) approach. The results show that the MOEs resulting from daily dietary intake did not raise a concern for endpoints for neurotoxicity including mitochondrial function, cytotoxicity, and apoptosis, while those for DNA adduct formation could not exclude a concern over genotoxicity. Endogenous MGO formation, especially under diabetic conditions, resulted in MOEs that raised concern not only for genotoxicity but also for some of the neurotoxicity endpoints evaluated. Thus, the results also point to the importance of taking the endogenous levels into account in the risk assessment of MGO.

Xyloglucan endotransglucosylase-hydrolase 1 is a negative regulator of drought tolerance in barley via modulating lignin biosynthesis and stomatal closure.

The projected increase in drought severity and duration worldwide poses a significant threat to crop growth and sustainable food production. Xyloglucan endotransglucosylase/hydrolases (XTHs) family is essential in cell wall modification through the construction and restructuring of xyloglucan cross-links, but their role in drought tolerance and stomatal regulation is still illusive. We cloned and functionally characterized HvXTH1 using genetic, physiological, biochemical, transcriptomic and metabolomic approaches in barley. Evolutionary bioinformatics showed that orthologues of XTH1 was originated from Streptophyte algae (e.g. some species in the Zygnematales) the closest clade to land plants based on OneKP database. HvXTH1 is highly expressed in leaves and HvXTH1 is localized to the plasma membrane. Under drought conditions, silencing HvXTH1 in drought-tolerant Tibetan wild barley XZ5 induced a significant reduction in water loss rate and increase in biomass, however overexpressing HvXTH1 exhibited drought sensitivity with significantly less drought-responsive stomata, lower lignin content and a thicker cell wall. Transcriptome profile of the wild type Golden Promise and HvXTH1-OX demonstrated that drought-induced differentially expressed genes in leaves are related to cell wall biosynthesis, abscisic acid and stomatal signaling, and stress response. Furthermore, overexpressing HvXTH1 suppressed both genes and metabolites in the phenylpropanoid pathway for lignin biosynthesis, leading to drought sensitivity of HvXTH1-OX. We provide new insight by deciphering the function of a novel protein HvXTH1 for drought tolerance in cell wall modification, stomatal regulation, and phenylpropanoid pathway for lignin biosynthesis in barley. The function of HvXTH1 in drought response will be beneficial to develop crop varieties adapted to drought.

Identification of a potent SARS-CoV-2 neutralizing nanobody targeting the receptor-binding domain of the spike protein.

SARS-CoV-2 and its variants continue to pose a significant threat to public health. Nanobodies (Nbs) that inhibit the interaction between the receptor-binding domain (RBD) of the spike protein and the host cell receptor angiotensin-converting enzyme 2 (ACE2) are promising drug candidates. In this study, we report the discovery and structural characterization of a potent Nb that targets the RBD. By screening a phage display alpaca naive Nbs library using the RBD as bait, we identified sixteen candidate Nbs. Of these, nine exhibited nanomolar to micromolar binding affinity and strong neutralizing activity against pseudotyped SARS-CoV-2 viruses, with NbS4 showing the highest neutralization potency. The crystal structure of the SARS-CoV-2 RBD in complex with NbS4 revealed that this Nb binds to a site partially overlapping the ACE2 binding region. Importantly, the key binding residues of NbS4 in the RBD are conserved across most known variants, making it a promising candidate for COVID-19 treatment.

Loss of NAT10 Reduces the Translation of Kmt5a mRNA Through ac4C Modification in Cardiomyocytes and Induces Heart Failure.

BACKGROUND: In the past decade, the biological functions of various RNA modifications in mammals have been uncovered. N4-acetylcytidine (ac4C), a highly conserved RNA modification, has been implicated in human diseases. Despite this, the involvement of RNA ac4C modification in cardiac physiology and pathology remains incompletely understood. NAT10 (N-acetyltransferase 10) stands as the sole acetyltransferase known to catalyze RNA ac4C modification. This study aims to explore the role of NAT10 and ac4C modification in cardiac physiology and pathology. METHODS AND RESULTS: Cardiac-specific knockout of NAT10, leading to reduced RNA ac4C modification, during both neonatal and adult stages resulted in severe heart failure. NAT10 deficiency induced cardiomyocyte apoptosis, a crucial step in heart failure pathogenesis, supported by in vitro data. Activation of the p53 signaling pathway was closely associated with enhanced apoptosis in NAT10-deficient cardiomyocytes. As ac4C modification on mRNA influences translational efficiency, we employed ribosome footprints coupled with RNA sequencing to explore genome-wide translational efficiency changes caused by NAT10 deficiency. We identified and validated that the translational efficiency of Kmt5a was suppressed in NAT10 knockout hearts due to reduced ac4C modification on its mRNA. This finding was consistent with the observation that Kmt5a protein levels were reduced in heart failure despite unchanged mRNA expression. Knockdown of Kmt5a in cardiomyocytes recapitulated the phenotype of NAT10 deficiency, including increased cardiomyocyte apoptosis and activated p53 signaling. Finally, overexpression of Kmt5a rescued cardiomyocyte apoptosis and p53 activation induced by NAT10 inhibition. CONCLUSIONS: Our study highlights the significance of NAT10 in cardiomyocyte physiology, demonstrating that NAT10 loss is sufficient to induce cardiomyocyte apoptosis and heart failure. NAT10 regulates the translational efficiency of Kmt5a, a key mediator, through mRNA ac4C modification during heart failure.

Metformin-mediated protection against doxorubicin-induced cardiotoxicity.

BACKGROUND: A phase II clinical trial of metformin (MET) for the treatment of doxorubicin (DOX)-induced cardiotoxicity (NCT02472353) failed. OBJECTIVES: The aims of this study were to confirm MET-mediated protection against DOX-induced cardiotoxicity and its mechanism using H9C2 cells, and to establish a Wistar rat model of DOX-induced cardiotoxicity. Subsequently, Wistar rats were utilized to identify clinically relevant indicators for evaluating MET-mediated protection against DOX-induced cardiotoxicity, thereby facilitating early transition towards successful clinical trials. METHODS: MET-mediated protection was assessed using cell viability and cytotoxicity experiments. Additionally, intramitochondrial reactive oxygen species (ROS) levels were measured using an ROS fluorescent probe (dihydroethidium) to confirm the oxidative stress mechanism. Eighteen Wistar rats were randomly allocated to the control, DOX, and DOX+MET groups; and the body weight, adverse drug reactions (ADRs), myocardial injury, cardiac function, oxidative stress, and histopathology of heart tissues were compared between groups. RESULTS: H9C2 cells treated with MET/Dexrazoxane demonstrated dose-dependent protection against DOX-induced cardiotoxicity. The fluorescence intensity of H9C2 cells suggested DOX-induced cardiomyocyte toxicity and MET-mediated protection against DOX-induced cardiotoxicity. In vivo experiments confirmed that a rat model of DOX-induced cardiotoxicity was successfully established, but MET-mediated protection against DOX-induced cardiotoxicity was not demonstrated. This was attributed to insufficient energy intake because of ADRs, such as vomiting. CONCLUSIONS: We confirmed the MET-mediated protection against DOX-induced cardiomyocyte toxicity and its mechanism involving the inhibition of oxidative stress in vitro experiments. It is imperative to investigate the optimal conditions for MET-mediated protection against DOX-induced cardiotoxicity in vivo or clinical trials.

Association of allostatic load with depression, anxiety, and suicide: a prospective cohort study.

BACKGROUND: Allostatic load (AL) is an objective measure of the biological components of chronic stress within clinical practice, potentially influencing depression, anxiety, and suicide. This study investigates the association between AL and these mental disorders. METHODS: In this cohort study of 333,017 adults, participants without prior diagnoses of depression, anxiety, or suicide were observed from March 13, 2006, to October 31, 2022. AL was estimated using 10 biomarkers reflecting metabolic, cardiovascular, and inflammatory dysregulation. Diagnoses were based on the International Classification of Diseases 10th Revision (ICD-10). We performed Cox proportional hazards models to assess the relationship between AL and these mental disorders. Additionally, we conducted subgroup analyses for sex, age, and Townsend Deprivation Index (TDI), along with sensitivity analyses. RESULTS: The median follow-up period was 12.99 years. Over the follow-up period, 13,441 (4.04%) participants developed depression, 13,903 (4.17%) developed anxiety and 796 (0.24%) committed suicide. In fully adjusted model, individuals with high AL had an increased risk of depression (HR = 1.389, P = 8.38 ×10-27), anxiety (HR = 1.304, P = 5.82 ×10-19) and suicide (HR = 1.433, P = 4.46 ×10-3). Women and younger individuals with high AL were vulnerable to depression and anxiety, while moderate AL levels were significantly associated with suicide in men and younger participants. Moreover, individuals with middle and high AL had an elevated risk of comorbid depression and anxiety. CONCLUSIONS: High AL is positively associated with increased risks of depression, anxiety, and suicide, highlighting its potential as a predictive tool in mental health.