pH-Universal Electrocatalytic CO2 Reduction with Ampere-level Current Density on Doping-engineered Bismuth Sulfide.

The practical application of the electrocatalytic CO2 reduction reaction (CO2RR) to form formic acid fuel is hindered by the limited activation of CO2 molecules and the lack of universal feasibility across different pH levels. Herein, we report a doping-engineered bismuth sulfide pre-catalyst (BiS-1) that S is partially retained after electrochemical reconstruction into metallic Bi for CO2RR to formate/formic acid with ultrahigh performance across a wide pH range. The best BiS-1 maintains a Faraday efficiency (FE) of ~95% at 2000 mA cm-2 in a flow cell under neutral and alkaline solutions. Furthermore, the BiS-1 catalyst shows unprecedentedly high FE (~95%) with current densities from 100 to 1300 mA cm-2 under acidic solutions. Notably, the current density can reach 700 mA cm-2 while maintaining a FE of above 90% in a membrane electrode assembly electrolyzer and operate stably for 150 h at 200 mA cm-2. In-situspectra and density functional theory calculations reveals that the S doping modulates the electronic structure of Bi and effectively promotes the formation of the HCOO* intermediate for formate/formic acid generation. This work develops the efficient and stable electrocatalysts for sustainable formate/formic acid production.

Effects of probiotic supplementation on 12 min run performance, mood management, body composition and gut microbiota in amateur marathon runners: A double-blind controlled trial.

Background: Probiotic supplementation has a positive effect on endurance exercise performance and body composition in athletes, but the underlying mechanisms remain unclear. Gut microbiota can provide measurable markers of immune function in athletes, and microbial composition analysis may be sensitive enough to detect stress and metabolic disorders caused by exercise. Methods: Nineteen healthy active amateur marathon runners (15 male and 4 female) with a mean age of 29.11 years volunteered to participate in this double-blind controlled study. Based on the performance of the Cooper 12-min running test (CRT), the participants were allocated into two groups to receive either a probiotic formulation comprising lactobacillus acidophilus and bifidobacterium longum (n = 10) or placebo containing maltodextrin (n = 9) for five weeks. Consistency of diet and exercise was ensured throughout the experimental period. Before and after the intervention, all participants were assessed for CRT, emotional stability and gastrointestinal symptoms, gut microbiota composition, body composition and magnetic resonance imaging (MRI) indicators of skeletal muscle microcirculation. Results: Compared to before the intervention, the probiotics group showed an increase in CRT score (2.88 ± 0.57 vs 3.01 ± 0.60 km, P＜0.05), significant improvement in GSRS and GIQLI (9.20 ± 4.64 vs 7.40 ± 3.24, 118.90 ± 12.30 vs 127.50 ± 9.85, P＜0.05), while these indicators remained unchanged in the control group, with a significant time-group interaction effect on gastrointestinal symptoms. Additionally, some MRI metabolic cycling indicators of the thigh skeletal muscle also changed in the probiotics group (P＜0.05). Regarding microbiota abundance, the probiotics group exhibited a significant increase in the abundance of beneficial bacteria and a significant decrease in the abundance of harmful bacteria post-intervention (P＜0.05). Conclusion: As a sports nutritional supplement, probiotics have the potential to improve athletic performance by optimizing the balance of gut microbiota, alleviating gastrointestinal symptoms.

Photocatalytic oxidation of formaldehyde under visible light using BiVO4-TiO2 synthesized via ultrasonic blending.

Formaldehyde (HCHO) is one of the primary indoor air pollutants, and efficiently eliminating it, especially at low concentrations, remains challenging. In this study, BiVO4-TiO2 catalyst was developed using ultrasonic blending technology for the photocatalytic oxidation of low-level indoor HCHO. The crystal structure, surface morphology, element distribution, and active oxidation species of the catalyst were examined using XRD, SEM, TEM, UV-Vis, EDS, and ESR techniques. Our results demonstrated that the BiVO4-TiO2 catalyst, prepared by ultrasonic blending, exhibited good oxidation performance and stability. The HCHO concentration reduced from 1.050 to 0.030 mg/m3 within 48 h, achieving a removal rate of 97.1%. The synergy between BiVO4 and TiO2 enhanced the efficiency of separating photogenerated carriers and minimized the likelihood of recombination between photogenerated electrons and holes. Additionally, this synergy significantly enhanced the presence of hydroxyl radicals (·OH) on the catalyst, resulting in an oxidation performance superior to that of either BiVO4 or TiO2. Our research offers valuable insights for the development of new photocatalysts to address HCHO pollution.

Boron in the Second Coordination Sphere of Fe Single Atom Boosts the Oxygen Reduction Reaction.

Metal single atoms coordinated with four nitrogen atoms (M1N4) are regarded as tremendously promising catalysts for the electrocatalytic oxygen reduction reaction (ORR). Nevertheless, the strong bond intensity between the metal center and the O atom in oxygen-containing intermediates significantly limits the ORR activity of M1N4. Herein, the catalytically active B atom is successfully introduced into the second coordination sphere of the Fe single atom (Fe1N4-B-C) to realize the alternative binding of B and O atoms and thus facilitate the ORR activity. Compared with the pristine Fe1N4 catalyst, the synthesized Fe1N4-B-C catalyst exhibits improved ORR catalytic capability with a half-wave potential (E1/2) of 0.80 V and a kinetic current density (JK) of 5.32 mA cm-2 in acid electrolyte. Moreover, in an alkaline electrolyte, the Fe1N4-B-C catalyst displays remarkable ORR activity with E1/2 of 0.87 V and JK of 8.94 mA cm-2 at 0.85 V, outperforming commercial Pt/C. Notably, the mechanistic study has revealed that the active center is the B atom in the second coordination shell of the Fe1N4-B-C catalyst, which avoids the direct bonding of Fe-O. The B center has a moderate binding force to the ORR intermediate, which flattens the ORR energy diagram and thereby improves the ORR performance. Therefore, this study offers a novel strategy for tailoring catalytic performance by tuning the active center of single-atom catalyst.

A replacement strategy for regulating local environment of single-atom Co-SxN4-x catalysts to facilitate CO2 electroreduction.

The performances of single-atom catalysts are governed by their local coordination environments. Here, a thermal replacement strategy is developed for the synthesis of single-atom catalysts with precisely controlled and adjustable local coordination environments. A series of Co-SxN4-x (x = 0, 1, 2, 3) single-atom catalysts are successfully synthesized by thermally replacing coordinated N with S at elevated temperature, and a volcano relationship between coordinations and catalytic performances toward electrochemical CO2 reduction is observed. The Co-S1N3 catalyst has the balanced COOH*and CO* bindings, and thus locates at the apex of the volcano with the highest performance toward electrochemical CO2 reduction to CO, with the maximum CO Faradaic efficiency of 98 ± 1.8% and high turnover frequency of 4564 h-1 at an overpotential of 410 mV tested in H-cell with CO2-saturated 0.5 M KHCO3, surpassing most of the reported single-atom catalysts. This work provides a rational approach to control the local coordination environment of the single-atom catalysts, which is important for further fine-tuning the catalytic performance.

Chemically Tailored Single Atoms for Targeted and Light-Controlled Bactericidal Activity.

Single-atom (SA) nanoparticles exhibit considerable potential in terms of photothermal properties for bactericidal applications. Nevertheless, the restricted efficacy of their targeted and controlled antibacterial activity has hindered their practical implementation. This study aims to overcome this obstacle by employing chemical modifications to tailor SAs, thereby achieving targeted and light-controlled antimicrobial effects. By conducting atomic-level modifications on palladium SAs using glutathione (GSH) and mercaptophenylboronic acid (MBA), their superior targeted binding capabilities toward Escherichia coli cells are demonstrated, surpassing those of SAs modified with cysteine (Cys). Moreover, these modified SAs effectively inhibit wound bacteria proliferation and promote wound healing in rats, without inducing noticeable toxicity to major organs under 808 nm laser irradiation. This study highlights the significance of chemical engineering in tailoring the antibacterial properties of SA nanoparticles, opening avenues for combating bacterial infections and advancing nanoparticle-based therapies.

Population impact of fine particulate matter on tuberculosis risk in China: a causal inference.

BACKGROUND: Previous studies have suggested the potential association between air pollution and tuberculosis incidence, but this association remains inconclusive and evidence to assess causality is particularly lacking. We aimed to draw causal inference between fine particulate matter less than 2.5 µm in diameter (PM2.5) and tuberculosis in China. METHODS: Granger causality (GC) inference was performed within vector autoregressive models at levels and/or first-differences using annual national aggregated data during 1982-2019, annual provincial aggregated data during 1982-2019 and monthly provincial aggregated data during 2004-2018. Convergent cross-mapping (CCM) approach was used to determine the backbone nonlinear causal association based on the monthly provincial aggregated data during 2004-2018. Moreover, distributed lag nonlinear model (DLNM) was applied to quantify the causal effects. RESULTS: GC tests identified PM2.5 driving tuberculosis dynamics at national and provincial levels in Granger sense. Empirical dynamic modeling provided the CCM causal intensity of PM2.5 effect on tuberculosis at provincial level and demonstrated that PM2.5 had a positive effect on tuberculosis incidence. Then, DLNM estimation demonstrated that the PM2.5 exposure driven tuberculosis risk was concentration- and time-dependent in a nonlinear manner. This result still held in the multi-pollutant model. CONCLUSIONS: Causal inference showed that PM2.5 exposure driving tuberculosis, which showing a concentration gradient change. Air pollutant control may have potential public health benefit of decreasing tuberculosis burden.

A universal approach to dual-metal-atom catalytic sites confined in carbon dots for various target reactions.

Here, a molecular-design and carbon dot-confinement coupling strategy through the pyrolysis of bimetallic complex of diethylenetriamine pentaacetic acid under low-temperature is proposed as a universal approach to dual-metal-atom sites in carbon dots (DMASs-CDs). CDs as the "carbon islands" could block the migration of DMASs across "islands" to achieve dynamic stability. More than twenty DMASs-CDs with specific compositions of DMASs (pairwise combinations among Fe, Co, Ni, Mn, Zn, Cu, and Mo) have been synthesized successfully. Thereafter, high intrinsic activity is observed for the probe reaction of urea oxidation on NiMn-CDs. In situ and ex situ spectroscopic characterization and first-principle calculations unveil that the synergistic effect in NiMn-DMASs could stretch the urea molecule and weaken the N-H bond, endowing NiMn-CDs with a low energy barrier for urea dehydrogenation. Moreover, DMASs-CDs for various target electrochemical reactions, including but not limited to urea oxidation, are realized by optimizing the specific DMAS combination in CDs.

Correlating the valence state of a Cu-based electrocatalyst for CO2 reduction to C2.

In this study, a facile ligand-protected strategy for preparing Cu@Cu2O and CuO nanoparticles is presented. The electrocatalyst efficacy of the CuO variant, particularly for CO2 reduction to multi-carbon products (C2+), is significant, boasting faradaic efficiencies (FEs) surpassing 85% and a current density peak at 340 mA cm-2. This exceptional performance markedly exceeds that of the Cu@Cu2O electrocatalyst. This observed enhancement in the electrosynthesis efficiency of C2+ is attributed to the abundant Cu0 active sites, which originate from the in situ electroreduction of CuO.

Distinct dual enzyme-like activities of Fe-N-C single-atom nanozymes enable discriminative detection of cellular glutathione.

Fe-N-C single-atom nanozymes readily achieved discriminative detection of glutathione (GSH) over other biothiols with similar structure due to the difference between POD-like and OXD-like activities regarding the kind of reactive oxygen species. This colorimetric sensor demonstrated the heterogeneity of GSH levels in different cells and accurately monitored cellular GSH fluctuation.

Acute effects of foam rolling and dynamic stretching on angle-specific change of direction ability, flexibility and reactive strength in male basketball players.

The purpose of our study was to determine the acute effects of dynamic stretching (DS), foam rolling (FR) and foam rolling combined with dynamic stretching (Combo) protocols on angle-specific change of direction (COD) ability, drop jump (DJ) performance and flexibility. Using a counterbalance crossover study design, eleven male basketball collegiate players (20.7 ± 0.6 years) were randomly assigned to one of the four protocols - control (CON), DS, FR, Combo - for each session, for a total of four sessions. A more aggressive foam cylinder with raised nodules, which is thought to be effective in stimulating the deep layer of muscle tissue, was used to observe for changes in their performance during sit and reach (SAR), DJ and COD tasks in 45 and 180 degrees. One-way repeated measures ANOVA was used to identify differences of each variable separately between interventions. The SAR after three interventions compared to the CON was significantly improved (F (3,30) = 5.903, P = 0.003, η2 = 0.371). In the 505 test, both limbs failed to show a significant improvement in COD deficit. The non-dominant limb showed a significant improvement of 6.4% after FR when performing the Y-shaped agility (F (3,30) = 4.962, P = 0.0065 < 0.05, η2 = 0.332). In the DJ, the reactive strength index and contact time changed significantly by 17.5% and -17.5% (η2 = 0.518, η2 = 0.571), respectively, immediately after FR. The current research suggested that FR may have an enhancing effect on COD speed in a 45° cutting task and neuromuscular function, while having the potential to improve non-dominant limb deficits in both COD tasks. In contrast, the Combo warm-up protocol did not produce a cumulative effect, suggesting the need for coaches to remain cautious about excessive warm-up duration.

Association between long-term exposure to ambient air pollutants and the risk of tuberculosis: A time-series study in Nantong, China.

Background: Increasing evidence has shown that the risk of tuberculosis (TB) might be related to the exposure to air pollutants; however, the findings are inconsistent and studies on long-term air pollutant exposure and TB risk are scarce. This study aime to assess the relationship between monthly exposure to air pollution and TB risk in Nantong, China. Methods: We collected the time series data on the number of TB cases, as well as environmental and socioeconomic covariates from January 2005 to December 2020. The impact of air pollutant exposure on TB risk was evaluated using the distributed lag nonlinear model (DLNM). Stratified analyses were conducted to examine the effect modifications of sex and age on the association between air pollutants and TB risk. Sensitivity analyses were applied to test the stability of the model. Results: There were a total of 54,096 cases of TB in Nantong during the study period. In the single-pollutant model, for each 10 µg/m3 increase in concentration, the pooled relative risks (RRs) of TB reached the maximum to 1.10 (95% confidence interval (CI): 1.04-1.16, lag 10 months) for particulate matter with aerodynamic diameter less than 2.5 µm (PM2.5), 1.05 (95% CI: 1.01-1.10, lag 9 months) for particulate matter with aerodynamic diameter less than 10 µm (PM10), and 1.11 (95%CI: 1.04-1.19, lag 10 months) for nitrogen dioxide (NO2). Ozone (O3) did not show significant effect on TB risk. Effect modifications of sex and age on the association between air pollutants and TB risk were not observed. The multi-pollutant model results showed no significant variation compared with the single-pollutant model. Conclusions: Our study suggests that air pollutants pose a substantial threat to the TB risk. Reducing air pollution might be crucial for TB prevention and control.

D-Band Center Optimization of Edge-Rich Ultrathin RuZn Nanosheets With Moiré Superlattices for pH-Universal Hydrogen Evolution Reaction.

Electrocatalytic hydrogen evolution reaction (HER) is a promising way to produce pure and clean hydrogen. However, the preparation of efficient and economical catalysts for pH-universal HER remains a challenging but rewarding task. Herein, ultrathin RuZn nanosheets (NSs) with moiré superlattices and abundant edges are synthesized. The RuZn NSs with unique structure exhibit superb HER performance with overpotentials of 11, 13, and 29 mV to achieve 10 mA cm-2 in 1 M KOH, 1 M PBS, and 0.5 M H2 SO4 , respectively, which is substantially lower than those of Ru NSs and RuZn NSs without moiré superlattices. Density functional theory investigations reveal that the charge transfer from Zn to Ru will lead the appropriate downshift of the d-band center of surface Ru atoms, thus accelerating hydrogen desorption from the Ru sites, lowering the dissociation energy barrier of water and greatly improving the HER performance. This work provides an effective design scheme for high-performance HER electrocatalysts over a wide pH range, and propose a general route to prepare Ru-based bimetallic nanosheets with moiré superlattices.

Combined impacts of environmental and socioeconomic covariates on HFMD risk in China: A spatiotemporal heterogeneous perspective.

BACKGROUND: Understanding geospatial impacts of multi-sourced influencing factors on the epidemic of hand-foot-and-mouth disease (HFMD) is of great significance for formulating disease control policies tailored to regional-specific needs, yet the knowledge is very limited. We aim to identify and further quantify the spatiotemporal heterogeneous effects of environmental and socioeconomic factors on HFMD dynamics. METHODS: We collected monthly province-level HFMD incidence and related environmental and socioeconomic data in China during 2009-2018. Hierarchical Bayesian models were constructed to investigate the spatiotemporal relationships between regional HFMD and various covariates: linear and nonlinear effects for environmental covariates, and linear effects for socioeconomic covariates. RESULTS: The spatiotemporal distribution of HFMD cases was highly heterogeneous, indicated by the Lorenz curves and the corresponding Gini indices. The peak time (R2 = 0.65, P = 0.009), annual amplitude (R2 = 0.94, P<0.001), and semi-annual periodicity contribution (R2 = 0.88, P<0.001) displayed marked latitudinal gradients in Central China region. The most likely cluster areas for HFMD were located in south China (Guangdong, Guangxi, Hunan, Hainan) from April 2013 to October 2017. The Bayesian models achieved the best predictive performance (R2 = 0.87, P<0.001). We found significant nonlinear associations between monthly average temperature, relative humidity, normalized difference vegetation index and HFMD transmission. Besides, population density (RR = 1.261; 95%CI, 1.169-1.353), birth rate (RR = 1.058; 95%CI, 1.025-1.090), real GDP per capita (RR = 1.163; 95%CI, 1.033-1.310) and school vacation (RR = 0.507; 95%CI, 0.459-0.559) were identified to have positive or negative effects on HFMD respectively. Our model could successfully predict months with HFMD outbreaks versus non-outbreaks in provinces of China from Jan 2009 to Dec 2018. CONCLUSIONS: Our study highlights the importance of refined spatial and temporal data, as well as environmental and socioeconomic information, on HFMD transmission dynamics. The spatiotemporal analysis framework may provide insights into adjusting regional interventions to local conditions and temporal variations in broader natural and social sciences.

Population-level health and economic impacts of introducing Vaccae vaccination in China: a modelling study.

INTRODUCTION: Given the ageing epidemic of tuberculosis (TB), China is facing an unprecedented opportunity provided by the first clinically approved next-generation TB vaccine Vaccae, which demonstrated 54.7% efficacy for preventing reactivation from latent infection in a phase III trial. We aim to assess the population-level health and economic impacts of introducing Vaccae vaccination to inform policy-makers. METHODS: We evaluated a potential national Vaccae vaccination programme in China initiated in 2024, assuming 20 years of protection, 90% coverage and US$30/dose government contract price. An age-structured compartmental model was adapted to simulate three strategies: (1) no Vaccae; (2) mass vaccination among people aged 15-74 years and (3) targeted vaccination among older adults (60 years). Cost analyses were conducted from the healthcare sector perspective, discounted at 3%. RESULTS: Considering postinfection efficacy, targeted vaccination modestly reduced TB burden (~20%), preventing cumulative 8.01 (95% CI 5.82 to 11.8) million TB cases and 0.20 (0.17 to 0.26) million deaths over 2024-2050, at incremental cost-effectiveness ratio of US$4387 (2218 to 10 085) per disability adjusted life year averted. The implementation would require a total budget of US$22.5 (17.6 to 43.4) billion. In contrast, mass vaccination had a larger bigger impact on the TB epidemic, but the overall costs remained high. Although both preinfection and postinfection vaccine efficacy type might have a maximum impact (>40% incidence rate reduction in 2050), it is important that the vaccine price does not exceed US$5/dose. CONCLUSION: Vaccae represents a robust and cost-effective choice for TB epidemic control in China. This study may facilitate the practice of evidence-based strategy plans for TB vaccination and reimbursement decision making.

Fibulin-2 Facilitates Malignant Progression of Hepatocellular Carcinoma.

BACKGROUND: Identification of biomarkers to assist in the clinical management of hepatocellular carcinoma represents an urgent requirement. Fibulin-2 is known to contribute to the development and progression of various cancer types. This research investigated the role of fibulin-2 in hepatocellular carcinoma and explored the possible mechanisms. METHODS: The expression of fibulin-2 in hepatocellular carcinoma was measured by bioinformatic analysis and confirmed by western blot and immunohistochemical staining in cell lines or patients' samples. The clinicopathologic features of hepatocellular carcinoma patients was analyzed. Cell viability assays were used to explore the role of fibulin-2 on proliferation in hepatocellular carcinoma. Western blot was conducted to uncover changes of protein expression of Ras-MEK-ERK1/2 pathway when Fibulin-2 was overexpressed or silenced. Flow cytometry analyses were used to determine the roles of fibulin-2 in the function of apoptosis and cell cycle. Subcutaneous xenograft mouse models showed the tumor growth pattern after fibulin-2 silence in vivo. RESULTS: We reported the upregulation of fibulin-2 in most hepatocellular carcinoma tissues and cells lines. Fibulin-2 promoted the proliferation of hepatocellular carcinoma cells in vitro by regulating Ras-MEK-ERK1/2 signaling pathway, whereas knockdown of fibulin-2 incurred the opposite effect on proliferation. Consistently, knockdown of fibulin-2 resulted in increased apoptosis and induced growth arrest during the G0/G1 phase transition. In vivo xenograft assessment confirmed that knockdown of fibulin-2 inhibited hepatocellular carcinoma tumor growth. CONCLUSIONS: Fibulin-2 exhibited tumor promotor activities in malignant progression of hepatocellular carcinoma. The results of the study highlighted the potential of fibulin-2 to be utilized as a promising biomarker and therapeutic target for hepatocellular carcinoma.

Competition and Neutralization: Thermally Induced Crystallization and Phase Evolution of Amorphous Calcium Phosphate with Cosubstitution of Larger and Smaller Divalent Cations.

The presented study evaluated the effect of cosubstitution of larger and smaller divalent cations on the thermally induced crystallization of amorphous calcium phosphate (ACP). The predesigned combinations of larger (Sr2+ and Ba2+) and smaller (Mg2+, Cu2+, and Co2+) divalent cations were carried out and their effects on the thermodynamic equilibrium between α/ß-tricalcium phosphate (TCP) were outlined. The coexistence of larger and smaller divalent cations shielded the formation of α-TCP and shifted the thermodynamic equilibrium toward the ß-TCP, which implied that the smaller cations dominated the crystalline phase. However, the retarded crystallization induced by the larger cations still remained and allowed ACP to maintain its amorphous nature partly or completely until a higher temperature.

Stretchable Oxygen-Tolerant Sensor Based on a Single-Atom Fe-N4 Electrocatalyst for Observing the Role of Oxidative Stress in Hypertension.

Oxidative stress and related oxidative damage have a causal relation with the pathogenesis of hypertension. Therefore, it is crucial to determine the mechanism of oxidative stress in hypertension by applying mechanical forces on cells to simulate hypertension while monitoring the release of reactive oxygen species (ROS) from cells under an oxidative stress environment. However, cellular level research has rarely been explored because monitoring the ROS released by cells is still challenging owing to the interference of O2. In this study, an Fe single-atom-site catalyst anchored on N-doped carbon-based materials (Fe SASC/N-C) was synthesized, which exhibits excellent electrocatalytic activity for the reduction of hydrogen peroxide (H2O2) at a peak potential of +0.1 V and can effectively avoid the interference of O2. Furthermore, we constructed a flexible and stretchable electrochemical sensor based on the Fe SASC/N-C catalyst to study the release of cellular H2O2 under simulated hypoxic and hypertension conditions. Density functional theory calculations show that the highest transition state energy barrier from the oxygen reduction reaction (ORR), i.e., O2 to H2O, is 0.38 eV. In comparison, the H2O2 reduction reaction (HPRR) can be completed only by overcoming a lower energy barrier of 0.24 eV, endowing the HPRR to be more favorable on Fe SASC/N-C compared with the ORR. This study provided a reliable electrochemical platform for real-time investigation of H2O2-related underlying mechanisms of the hypertension process.

Identification of a CEACAM5 targeted nanobody for positron emission tomography imaging and near-infrared fluorescence imaging of colorectal cancer.

PURPOSE: Here, we aim to identify a CEACAM5-targeted nanobody and demonstrate its application in positron emission tomography (PET) imaging and near-infrared (NIR) fluorescence imaging in colorectal cancer (CRC). METHODS: Immunohistochemistry was applied to verify CEACAM5 expression in CRC and metastatic lymph nodes (mLNs). CEACAM5-targeted nanobodies were obtained by immunization of human CEACAM5 protein in a dromedary, followed by several rounds of phage screenings. Immunofluorescence staining and flow cytometry was carried out to determine the binding affinity of the nanobodies. The nanobodies were radiolabeled by coupling 18F-SFB for PET imaging of CRC subcutaneous xenografts and lymph node metastasis (LNM). IRDye800CW (IR800) were conjugated to form NIR probes for NIR imaging in CRC subcutaneous models. RESULTS: CEACAM5 was overexpressed in either human CRC tissues or mLNs. A CEACAM5 targeted nanobody, Nb41 was successfully generated, with excellent in vitro binding properties. Incorporation of albumin binding domain (ABD) did not affect the affinity of Nb41. In vivo imaging showed that both 18F-FB-Nb41 and 18F-FB-Nb41-ABD showed obvious accumulation in the tumor. Due to the longer retention in the blood, 18F-FB-Nb41-ABD enrichment in tumors was significantly delayed but higher compared to 18F-FB-Nb41. Both 18F-FB-Nb41 and 18F-FB-Nb41-ABD showed prominent LNM enrichment. Similarly, the IR800-conjugated nanobodies Nb41-IR800 and Nb41-ABD-IR800 exhibited superior imaging effects in subcutaneous models, while Nb41-ABD-IR800 exhibited higher fluorescence intensity in the tumor accompanied with a remarkedly delay compared to Nb41-IR800. CONCLUSION: Collectively, we presented the identification and in vivo validation of a CEACAM5-targeted nanobody and a fused nanobody with an ABD, which enabled to the non-invasive visualization of malignancy of CRC using PET imaging and NIR imaging in subcutaneous models as well as LNM models.

Mitochondrial ROS driven by NOX4 upregulation promotes hepatocellular carcinoma cell survival after incomplete radiofrequency ablation by inducing of mitophagy via Nrf2/PINK1.

BACKGROUND: The recurrence of hepatocellular carcinoma (HCC) after radiofrequency ablation (RFA) remains a major clinical problem. Cells that survive the sublethal heat stress that is induced by incomplete RFA are the main source of HCC relapse. Heat stress has long been reported to increase intracellular reactive oxygen species (ROS) generation. Although ROS can induce apoptosis, a pro-survival effect of ROS has also been demonstrated. However, the role of ROS in HCC cells exposed to sublethal heat stress remains unclear. METHODS: HepG2 and HuH7 cells were used for this experiment. Insufficient RFA was performed in cells and in a xenograft model. ROS and antioxidant levels were measured. Apoptosis was analyed by Annexin-V/PI staining and flow cytometry. Protein expression was measured using western blotting. Colocalization of lysosomes and mitochondria was analyzed to assess mitophagy. Corresponding activators or inhibitors were applied to verify the function of specific objectives. RESULTS: Here,we showed that sublethal heat stress induced a ROS burst, which caused acute oxidative stress. This ROS burst was generated by mitochondria, and it was initiated by upregulated NOX4 expression in the mitochondria. N-acetylcysteine (NAC) decreased HCC cell survival under sublethal heat stress conditions in vivo and in vitro. NOX4 triggers the production of mitochondrial ROS (mtROS), and NOX4 inhibitors or siNOX4 also decreased HCC cell survival under sublethal heat stress conditions in vitro. Increased mtROS trigger PINK1-dependent mitophagy to eliminate the mitochondria that are damaged by sublethal heat stress and to protect cells from apoptosis. Nrf2 expression was elevated in response to this ROS burst and mediated the ROS burst-induced increase in PINK1 expression after sublethal heat stress. CONCLUSION: These data confirmed that the ROS burst that occurs after iRFA exerted a pro-survival effect. NOX4 increased the generation of ROS by mitochondria. This short-term ROS burst induced PINK1-dependent mitophagy to eliminate damaged mitochondria by increasing Nrf2 expression.