High-performance seawater oxidation by a homogeneous multimetallic layered double hydroxide electrocatalyst.

SignificanceSeawater is one of the most abundant resources on Earth. Direct electrolysis of seawater is a transformative technology for sustainable hydrogen production without causing freshwater scarcity. However, this technology is severely impeded by a lack of robust and active oxygen evolution reaction (OER) electrocatalysts. Here, we report a highly efficient OER electrocatalyst composed of multimetallic layered double hydroxides, which affords superior catalytic performance and long-term durability for high-performance seawater electrolysis. To the best of our knowledge, this catalyst is among the most active for OER and it advances the development of seawater electrolysis technology.

Confined Water Dominates Ion/Molecule Transport in Hydrogel Nanochannels.

Current artificial nanochannels rely more on charge interactions for intelligent mass transport. Nevertheless, popular charged nanochannels would lose their advantages in long-term applications. Confined water, an indispensable transport medium in biological nanochannels, dominating the transport process in the uncharged nanochannels perfectly provides a new perspective. Herein, we achieve confined-water-dominated mass transport in hydrogel nanochannels (HNCs) constructed by in situ photopolymerization of acrylic acid (PAA) hydrogel in anodic alumina (AAO) nanochannels. HNCs show selectivity to Na+ transport and a high transport rate of molecules after introducing Na+/Li+, compared with other alkali metal ions like Cs+/K+. The mechanism given by ATR-FTIR shows that the hydrogen-bonding structure of confined water in HNCs is destabilized by Na+/Li+, which facilitates mass transport, but is constrained by Cs+/K+, resulting in transport inhibition. This work elucidates the relationship between confined water and mass transport in uncharged nanochannels while also presenting a strategy for designing functional nanochannel devices.

Mitochondrial and metabolic dysfunction of peripheral immune cells in multiple sclerosis.

Multiple sclerosis (MS) is a chronic autoimmune disorder characterized by the infiltration of inflammatory cells and demyelination of nerves. Mitochondrial dysfunction has been implicated in the pathogenesis of MS, as studies have shown abnormalities in mitochondrial activities, metabolism, mitochondrial DNA (mtDNA) levels, and mitochondrial morphology in immune cells of individuals with MS. The presence of mitochondrial dysfunctions in immune cells contributes to immunological dysregulation and neurodegeneration in MS. This review provided a comprehensive overview of mitochondrial dysfunction in immune cells associated with MS, focusing on the potential consequences of mitochondrial metabolic reprogramming on immune function. Current challenges and future directions in the field of immune-metabolic MS and its potential as a therapeutic target were also discussed.

Genomic epidemiology of CVA10 in Guangdong, China, 2013-2021.

Hand, Foot and Mouth Disease (HFMD) is a highly contagious viral illness primarily affecting children globally. A significant epidemiological transition has been noted in mainland China, characterized by a substantial increase in HFMD cases caused by non-Enterovirus A71 (EV-A71) and non-Coxsackievirus A16 (CVA16) enteroviruses (EVs). Our study conducts a retrospective examination of 36,461 EV-positive specimens collected from Guangdong, China, from 2013 to 2021. Epidemiological trends suggest that, following 2013, Coxsackievirus A6 (CVA6) and Coxsackievirus A10 (CVA10) have emerged as the primary etiological agents for HFMD. In stark contrast, the incidence of EV-A71 has sharply declined, nearing extinction after 2018. Notably, cases of CVA10 infection were considerably younger, with a median age of 1.8 years, compared to 2.3 years for those with EV-A71 infections, possibly indicating accumulated EV-A71-specific herd immunity among young children. Through extensive genomic sequencing and analysis, we identified the N136D mutation in the 2 A protein, contributing to a predominant subcluster within genogroup C of CVA10 circulating in Guangdong since 2017. Additionally, a high frequency of recombination events was observed in genogroup F of CVA10, suggesting that the prevalence of this lineage might be underrecognized. The dynamic landscape of EV genotypes, along with their potential to cause outbreaks, underscores the need to broaden surveillance efforts to include a more diverse spectrum of EV genotypes. Moreover, given the shifting dominance of EV genotypes, it may be prudent to re-evaluate and optimize existing vaccination strategies, which are currently focused primarily target EV-A71.

A chimeric porcine reproductive and respiratory syndrome virus 1 strain containing synthetic ORF2-6 genes can trigger T follicular helper cell and heterologous neutralizing antibody responses and confer enhanced cross-protection.

The prevalence of porcine reproductive and respiratory syndrome virus 1 (PRRSV1) isolates has continued to increase in Chinese swine herds in recent years. However, no effective control strategy is available for PRRSV1 infection in China. In this study, we generated the first infectious cDNA clone (rHLJB1) of a Chinese PRRSV1 isolate and subsequently used it as a backbone to construct an ORF2-6 chimeric virus (ORF2-6-CON). This virus contained a synthesized consensus sequence of the PRRSV1 ORF2-6 gene encoding all the envelope proteins. The ORF2-6 consensus sequence shared > 90% nucleotide similarity with four representative strains (Amervac, BJEU06-1, HKEU16 and NMEU09-1) of PRRSV1 in China. ORF2-6-CON had replication efficacy similar to that of the backbone rHLJB1 virus in primary alveolar macrophages (PAMs) and exhibited cell tropism in Marc-145 cells. Piglet inoculation and challenge studies indicated that ORF2-6-CON is not pathogenic to piglets and can induce enhanced cross-protection against a heterologous SD1291 isolate. Notably, ORF2-6-CON inoculation induced higher levels of heterologous neutralizing antibodies (nAbs) against SD1291 than rHLJB1 inoculation, which was concurrent with a higher percentage of T follicular helper (Tfh) cells in tracheobronchial lymph nodes (TBLNs), providing the first clue that porcine Tfh cells are correlated with heterologous PRRSV nAb responses. The number of SD1291-strain-specific IFNγ-secreting cells was similar in ORF2-6-CON-inoculated and rHLJB1-inoculated pigs. Overall, our findings support that the Marc-145-adapted ORF2-6-CON can trigger Tfh cell and heterologous nAb responses to confer improved cross-protection and may serve as a candidate strain for the development of a cross-protective PRRSV1 vaccine.

A diet-wide association study for liver cancer risk: findings from a prospective cohort study in Chinese men.

Dietary factors have been extensively investigated as possible risk factors for liver cancer, but the evidence is inconclusive. Our study systematically assessed the association between 142 foods and nutrients and liver cancer risk in a Chinese population using a diet-wide association study. Based on data from 59,844 men in the Shanghai Men's Health Study (SMHS), we assessed the diet intake by dietary questionnaires. Cox regression was used to quantify the association between each food and nutrient and liver cancer risk. A false discovery rate (FDR) of 0.05 was used to select the foods and nutrients for validation. In the cohort, 431 liver cancer cases were identified during 712,373 person-years of follow-up. Retinol (HR per 1 SD increment = 1.09, 95% CI: 1.03-1.14) was associated with a higher risk of liver cancer, whereas onions (HR per 1 SD increment = 0.67, 95% CI: 0.54-0.84) and manganese (HR per 1 SD increment = 0.85, 95% CI: 0.78-0.94) were inversely associated with liver cancer risk. In the replication analysis, estimates for these foods and nutrients were similar in magnitude and direction. Our findings confirm that retinol, onions and manganese were associated with liver cancer risk, which provides reliable evidence between diet and liver cancer development.

Sex-specific impact of dietary patterns on liver cancer incidence: updated results from two population-based cohort studies in China.

PURPOSE: The associations between dietary patterns and liver cancer risk have received much attention, but evidence among the Chinese population is scarce. This study aims to update the results of two cohort studies and provide the sex-specific associations in the Chinese population. METHODS: This study was based on two cohorts from the Shanghai Men's Health Study (SMHS) and the Shanghai Women's Health Study (SWHS). Diet information was collected by validated food frequency questionnaires. Dietary patterns were derived by factor analysis. Cox regression model was utilized to estimate the hazard ratio (HR) and 95% confidence interval (CI) for associations between dietary patterns and liver cancer risk. RESULTS: During median follow-up years of 11.2 (male) and 17.1 (female) years, 427 males and 252 females were identified as incident primary liver cancer cases. In males, vegetable-based dietary pattern was inversely associated with liver cancer (HRQ4-Q1: 0.67, 95%CI 0.51-0.88, Ptrend < 0.001). Interaction analysis indicated that in males lower vegetable-based dietary pattern score and older age/medical history of chronic hepatitis combined increase the hazard of liver cancer more than the sum of them, with a 114% and 1061% higher risk, respectively. In females, the fruit-based dietary pattern was associated with a reduced risk of liver cancer (HRQ4-Q1: 0.63, 95%CI 0.42-0.95, Ptrend = 0.03). In both males and females, null associations were observed between the meat-based dietary pattern and the risk of liver cancer. CONCLUSION: A vegetable-based dietary pattern in males and a fruit-based dietary pattern in females tended to have a protective role on liver cancer risk. This study provided updated information that might be applied to guide public health action for the primary prevention of liver cancer.

Baicalein triggers ferroptosis in colorectal cancer cells via blocking the JAK2/STAT3/GPX4 axis.

Colorectal cancer (CRC) is a prevalent form of gastrointestinal malignancy with challenges in chemotherapy resistance and side effects. Effective and low toxic drugs for CRC treatment are urgently needed. Ferroptosis is a novel mode of cell death, which has garnered attention for its therapeutic potential against cancer. Baicalein (5, 6, 7-trihydroxyflavone) is the primary flavone extracted from the dried roots of Scutellaria baicalensis that exhibits anticancer effects against several malignancies including CRC. In this study, we investigated whether baicalein induced ferroptosis in CRC cells. We showed that baicalein (1-64 µM) dose-dependently inhibited the viability of human CRC lines HCT116 and DLD1. Co-treatment with the ferroptosis inhibitor liproxstatin-1 (1 µM) significantly mitigated baicalein-induced CRC cell death, whereas autophagy inhibitor chloroquine (25 µM), necroptosis inhibitor necrostatin-1 (10 µM), or pan-caspase inhibitor Z-VAD-FMK (10 µM) did not rescue baicalein-induced CRC cell death. RNA-seq analysis confirmed that the inhibitory effect of baicalein on CRC cells is associated with ferroptosis induction. We revealed that baicalein (7.5-30 µM) dose-dependently decreased the expression levels of GPX4, key regulator of ferroptosis, in HCT116 and DLD1 cells by blocking janus kinase 2 (JAK2)/STAT3 signaling pathway via direct interaction with JAK2, ultimately leading to ferroptosis in CRC cells. In a CRC xenograft mouse model, administration of baicalein (10, 20 mg/kg, i.g., every two days for two weeks) dose-dependently inhibited the tumor growth with significant ferroptosis induced by inhibiting the JAK2/STAT3/GPX4 axis in tumor tissue. This study demonstrates that ferroptosis contributes to baicalein-induced anti-CRC activity through blockade of the JAK2/STAT3/GPX4 signaling pathway, which provides evidence for the therapeutic application of baicalein against CRC.

Residential mobility and liver cancer risk: findings from a prospective cohort study in Chinese women.

BACKGROUND: Residential mobility is believed to influence the occurrence and development of cancer; however, the results are inconclusive. Furthermore, limited studies have been conducted on Asian populations. This study aimed to evaluate the relationship between residential mobility and liver cancer risk among Chinese women. METHODS: We enrolled 72,818 women from urban Shanghai between 1996 and 2000, and then followed them until the end of 2016. Cox regression models were used to calculate hazard ratios (HRs) and 95% confidence intervals (CIs) to assess the association between residential mobility and liver cancer risk. A linear trend test was conducted by ranking variables. A sensitivity analysis was also conducted, excluding participants with follow-up times of less than 2 years, to prevent potential bias. RESULTS: During the 1,269,765 person-years of follow-up, liver cancer was newly diagnosed in 259 patients. Domestic migration (HR = 1.47, 95% CI, 1.44-1.50), especially immigration to Shanghai (HR = 1.47, 95% CI, 1.44-1.50) was associated with an increased risk of liver cancer. In addition, migration frequency, age at initial migration and first immigration to Shanghai had linear trends with an increased liver cancer risk (Ptrend <0.001). The results were similar when excluding participants with less than two years of follow-up. CONCLUSIONS: The possible association between residential mobility and a higher risk of liver cancer in women could suggest the need for effective interventions to reduce adverse environmental exposures and enhance people's health.

Construction of lignocellulose-degrading compound microbial inoculum and its effects on green waste composting.

The high content of recalcitrant lignocellulose in green waste (GW) makes composting and degradation challenging. Conventional GW composting typically employs single-strain microbial inoculants (MIs) with limited enzyme production capabilities, resulting in low composting efficiency and suboptimal compost product quality. In this study, Bacillus amyloliquefaciens (J1), Clonostachys rogersoniana (B2), and Streptomyces thermoviolaceus (J3) was utilized to optimize cultivation conditions and strain ratios based on enzyme activity indicators. The aim was to develop a potent three-strain lignocellulose-degrading MIs and test the hypothesis that its performance is superior to that of single-strain and two-strain MIs in terms of lignocellulose degradation and compost maturation. The results indicated that, the optimal treatment was T7, which was inoculated with a three-strain MIs composed of the spore suspensions of J1, B2, and J3 with a volume ratio of 3:3:2. Specifically, compared to the control (without MI), T7 increased the content of particle size between 0.25 and 2.00 mm and humic acid by 17% and 291%, respectively. Furthermore, T7 enhanced the degradation rates of cellulose, hemicellulose, and lignin by 197%, 145%, and 113%, respectively, and increased the activities of laccase, manganese peroxidase, lignin peroxidase, and carboxymethyl cellulase by 605%, 269%, 180%, and 228%, respectively. Additionally, T7 increased the relative abundance of bacteria (e.g. Pseudomonas) and fungi (e.g. Parascedosporium) that facilitated lignocellulose degradation, enhanced the alpha diversity index and promoted the formation of a microbial community structure characterized by prominent dominant species and greater diversity. Remarkably, the inoculation with the three-strain MI yielded high-quality compost within 32 days.

Artemisinin and Its Derivatives as Potential Anticancer Agents.

Artemisinin is a natural sesquiterpene lactone obtained from the traditional Chinese medicinal herb Artemisia annua L. (qinghao). Artemisinin and its derivatives share an unusual endoperoxide bridge and are extensively used for malaria treatment worldwide. In addition to antimalarial activities, artemisinin and its derivatives have been reported to exhibit promising anticancer effects in recent decades. In this review, we focused on the research progress of artemisinin and its derivatives with potential anticancer activities. The pharmacological effects, potential mechanisms, and clinical trials in cancer therapy of artemisinin and its derivatives were discussed. This review may facilitate the future exploration of artemisinin and its derivatives as effective anticancer agents.

Regulating Spin Polarization via Axial Nitrogen Traction at Fe-N5 Sites Enhanced Electrocatalytic CO2 Reduction for Zn-CO2 Batteries.

Single Fe sites have been explored as promising catalysts for the CO2 reduction reaction to value-added CO. Herein, we introduce a novel molten salt synthesis strategy for developing axial nitrogen-coordinated Fe-N5 sites on ultrathin defect-rich carbon nanosheets, aiming to modulate the reaction pathway precisely. This distinctive architecture weakens the spin polarization at the Fe sites, promoting a dynamic equilibrium of activated intermediates and facilitating the balance between *COOH formation and *CO desorption at the active Fe site. Notably, the synthesized FeN5, supported on defect-rich in nitrogen-doped carbon (FeN5@DNC), exhibits superior performance in CO2RR, achieving a Faraday efficiency of 99% for CO production (-0.4 V vs. RHE) in an H-cell, and maintaining a Faraday efficiency of 98% at a current density of 270 mA cm-2 (-1.0 V vs. RHE) in the flow cell. Furthermore, the FeN5@DNC catalyst is assembled as a reversible Zn-CO2 battery with a cycle durability of 24 hours. In-situ IR spectroscopy and density functional theory (DFT) calculations reveal that the axial N coordination traction induces a transformation in the crystal field and local symmetry, therefore weakening the spin polarization of the central Fe atom and lowering the energy barrier for *CO desorption.

Large-Scale Metal-Organic Framework Nanoparticle Monolayers with Controlled Orientation for Selective Transport of Rare-Earth Elements.

Long-range ordered membranes comprised of porous nanoparticles have been pursued in precise separations for a long time. Yet most of the fabrication methods suffer from limited substrates or lack of precise control over crystal orientation. Herein, large-scale metal-organic framework (MOF) monolayer membranes with controlled orientations are prepared through an interfacial self-assembly process confined by superlyophilic substrates. The superspreading of reactant microdroplets results in an ultrathin liquid layer under an immiscible oil as a confined reactor. The concomitant MOF (ZIF-8) particles spontaneously assemble into monolayers with controlled orientations, determined by the particles' contact angles at the liquid/liquid interface, which can be regulated by solvent compositions. Therein both gas-adsorption and ion-transport tests prove that the ⟨111⟩-oriented membrane exhibits a minimized mass-transfer resistance. The as-prepared membrane can selectively transport rare-earth elements (REEs), and a La3+/K+ selectivity of 14.3 is achieved. Molecular dynamics simulations reveal that the REEs-selectivity is associated with the distinct difference in ion-membrane binding energies, demonstrating the potential of ZIF-8 membranes for use in high-efficiency recovery of REEs from industrial wastes.

Structural diversity, hypothetical biosynthesis, chemical synthesis, and biological activity of Ganoderma meroterpenoids.

Covering: 2018 to 2022Meroterpenoids found in fungal species of the genus Ganoderma and known as Ganoderma meroterpenoids (GMs) are substances composed of a 1,2,4-trisubstituted benzene and a polyunsaturated side chain. These substances have attracted the attention of chemists and pharmacologists due to their diverse structures and significant bioactivity. In this review, we present the structures and possible biosynthesis of representative GMs newly found from 2018 to 2022, as well as chemical synthesis and biological activity of some interesting GMs. We propose for the first time a plausible biosynthetic pathway for GMs, which will certainly motivate further research on the biosynthetic pathway in Ganoderma species, as well as on chemical synthesis of GMs as important bioactive compounds for the purpose of drug development.

Sex-Specific Correlation Analysis of Branched-Chain Amino Acids in Dietary Intakes and Plasma among Chinese Adults.

BACKGROUND: Previous findings about the influence of dietary intakes of the branched-chain amino acid (BCAA) on their plasma concentrations have been limited and inconsistent, and evidence from the Chinese population was lacking. OBJECTIVES: This study aimed to investigate the diet-plasma BCAA correlations in Chinese male and female adults. METHODS: This cross-sectional study was based on a nested case-control study within 2 prospective population-based cohorts in Shanghai, China. Diet information was collected by the food frequency questionnaires. Plasma BCAA concentrations were measured by ultraperformance liquid chromatography coupled with tandem mass spectrometry. Spearman correlations and linear regression models were conducted to examine the relationships between dietary BCAA intakes and plasma BCAA. The multivariable model was adjusted for age at the interview, total energy intake, time of blood collection from last meal, dietary patterns, body mass index (in kg/m2), type 2 diabetes, and physical activity. RESULTS: A total of 322 males (median age of 57.0 y) and 187 females (median age of 60.0 y) were included in this cross-sectional study. The geometric means of dietary intake of leucine, isoleucine, valine, and BCAA were 4937.7, 3029.6, 3268.5, and 11237.4 mg/d in males, and 4125.7, 2567.8, 2754.3, and 9449.4 mg/d in females. The geometric means of plasma concentrations of leucine, isoleucine, valine, and BCAA were 181.9, 65.0, 219.8, and 469.4 µM/L in males and 161.6, 61.1, 206.5, and 431.6 µM/L in females. Only leucine (r = 0.1660, P = 0.0028) and total BCAA (r = 0.1348, P = 0.0155) in males exhibited weak positive correlation coefficients. After adjustment for the covariates, leucine, isoleucine, valine, and total BCAA in dietary intakes and plasma were not correlated in both males and females. CONCLUSIONS: In Chinese male and female adults, dietary intakes are not major determinants of plasma concentrations of BCAA, and plasma concentrations might not be reflected by usual dietary intakes of BCAA.

Cross-sectional relationships between general and central adiposity and plasma amino acids in Chinese adults.

Adiposity is an important determinant of blood metabolites, but little is known about the variations of blood amino acids according to general and central adiposity status among Chinese population. This study included 187 females and 322 males who were cancer-free subjects randomly selected from two cohorts in Shanghai, China. Participants' plasma concentrations of amino acids were measured by ultra-performance liquid chromatography coupled to tandem mass spectrometry. Linear regression models were used to examine the cross-sectional correlations between general and central adiposity and amino acid levels. A total of 35 amino acids in plasma were measured in this study. In females, alanine, aspartic acid and pyroglutamic acid were positively correlated with general adiposity. In males, glutamic acid, aspartic acid, valine and pyroglutamic acid showed positive correlations, and glutamine, serine and glycine showed negative correlations with both general and central adiposity; phenylalanine, isoleucine and leucine were positively correlated and N-phenylacetylglutamine was negatively correlated with general adiposity; asparagine was negatively correlated with central adiposity. In summary, general adiposity and central adiposity were correlated with the concentrations of specific plasma amino acids among cancer-free female and male adults in China. Adiposity-metabolite characteristics and relationships should be considered when studying blood biomarkers for adiposity-related health outcomes.

Development of actein derivatives as potent anti-triple negative breast cancer agents.

Actein is a natural triterpenoid glycoside, isolated from the rhizomes of Cimicifuga foetida, which have been demonstrated to be potential in the treatment of breast cancer previously. Herein, we described the design and synthesis of a series of actein derivatives as anti-triple negative breast cancer (TNBC) inhibitors. Of which, the most promising derivative 27 exhibited significant inhibitory activity against human TNBC cell lines HCC1806 and MDA-MB-231, with IC50 values of 2.78 and 9.11 µM, respectively. Structure-activity relationships of actein derivatives were also discussed. Moreover, preliminary mechanism investigation revealed that 27 significantly inhibited cancer cell proliferation via cell cycle arrest at S phase. In addition, western blot analysis showed that the activation of MAPK signaling pathway might contribute to derivative 27 induced cell death. Overall, these results indicate that 27 has the potential to be developed as a lead compound and compounds with the actein scaffold are a promising novel class of inhibitors to treat TNBC.

Covalent Organic Framework/g-C3N4 van der Waals Heterojunction toward H2 Production.

Photocatalytic water splitting into H2 is the most economic and environmentally friendly strategy for H2 production, and rationally constructing a heterojunction retains enormous influence on a photocatalytic system. Herein, 2D/2D covalent organic framework/graphitic carbon nitride (COF/CN) van der Waals heterojunctions were readily prepared via an ultrasonic method for high-efficiency visible-light photocatalytic H2 production. The photocatalytic H2 production performance of optimized COF/CN composites can reach up to 449.64 µmol·h-1, which is approximately 5 times that of pure CN (89.08 µmol·h-1). The characterization and experimental studies reveal that the synergistic effect between COF and CN contributes to promoting the interfacial migration and spatial separation of photoinduced e--h+ pairs, further boosting the photocatalytic hydrogen production activity. This work may open a new window to design and fabricate effective heterojunction photocatalysts for photocatalytic energy conversion.

Association between the triglyceride-glucose index and albuminuria in hypertensive individuals.

BACKGROUND: Previous studies have demonstrated that the triglyceride-glucose (TyG) index is significantly associated with vascular damage. Albuminuria is a marker of hypertension-mediated organ damage (HMOD) and has been linked to a greater risk of cardiovascular disease (CVD). However, the association between the TyG index and albuminuria in patients with hypertension is not clear. This population research focused on subjects with hypertension to investigate the association between an elevated TyG index and albuminuria. METHODS: From September 2019 to November 2019, 789 hypertensive participants were involved in our research. Logistic regression models were performed to estimate the odds ratios (ORs) and 95% confidence intervals (CIs) for albuminuria according to the quartiles of the TyG index. RESULTS: Multivariate logistic regression analysis revealed that the TyG index was significantly associated with albuminuria. Using the lowest TyG index quartile as the reference, the fully adjusted ORs (95% CIs) for albuminuria for TyG index quartile II, quartile III, and quartile IV were 1.90 (1.17-3.12), 1.81 (1.07-3.07), and 3.46 (2.06-5.91), respectively. The results in the subgroup analysis were similar to the main analyses except for the smokers. Restricted cubic spline curves based on logistic regression models evaluated the linear association between the TyG index and albuminuria (P for nonlinear = 0.831). CONCLUSION: The TyG index was positively associated with albuminuria among hypertensive participants.

Interaction of SOX5 with SOX9 promotes warfarin-induced aortic valve interstitial cell calcification by repressing transcriptional activation of LRP6.

Calcific aortic valve disease (CAVD) is an important health burden due to its increasing prevalence and lack of available approaches. Osteogenic transdifferentiation of aortic valve interstitial cells (AVICs) contributes to valve calcification. SRY-related HMG-box transcription factor 5 (SOX5) is essential for cartilage development. Whether SOX5 is involved in AVIC calcification has not been determined. This study aimed to explore the role of SOX5 in warfarin-induced AVIC calcification. Immunostaining showed decreased SOX5 in human calcific AV and warfarin induced mouse calcific AV tissues compared with human noncalcific AV and control mouse AV tissues. In calcific human AVICs (hAVICs) and porcine AVICS (pAVICs), both knockdown and overexpression of SOX5 inhibited calcium deposition and osteogenic marker gene expression. Protein expression assays and ChIP assays showed that overexpression of SOX5 led to increased recruitment of SOX5 to the SOX9 promoter and resulted in increased mRNA and protein expression of SOX9. Coimmunoprecipitation and immunofluorescence showed that SOX5 binds to SOX9 with its HMG domain in nucleus. Blue Native PAGE showed overexpression of SOX5 led to multimeric complex formation of SOX5 and resulted in decreased binding of SOX5 to SOX9 similar to the results of knockdown of SOX5. Further ChIP and western blotting assays showed that both knockdown and overexpression of SOX5 resulted in SOX9 initiating transcription of anti-calcific gene LRP6 in warfarin-treated pAVICs. Knockdown of LRP6 rescues the anti-calcification effect of SOX5 overexpression. We found that both loss and gain of function of SOX5 lead to the same phenotype: decreased warfarin induced calcification. The stoichiometry of SOX5 is crucial for cooperation with SOX9, SOX9 nuclear localization and subsequent binding of SOX9 to LRP6 promoter. These results suggest that SOX5 is a potential target for the development of anti-calcification therapy.