Hepatitis B virus (HBV) infection remains a leading etiological driver of hepatocellular carcinoma (HCC). Cuproptosis is a recently defined copper-dependent form of regulated cell death that selectively eliminates mitochondria-dependent cells; whether HBV rewires this vulnerability remains unknown. Here we unveil a novel HBV X protein (HBx)-driven mechanism of cuproptosis evasion. Integrative analysis of clinical specimens, HBx-transgenic (HBx-Tg) mice, and multi-omics datasets revealed marked downregulation of STEAP4 (six-transmembrane epithelial antigen of prostate 4), a metalloreductase essential for cuproptosis sensitivity, in HBV-positive HCC. Mechanistically, HBx attenuates sirtuin 3 (SIRT3), impairing deacetylation of STEAP4 at lysine 404 and abolishing its mitochondrial targeting. Consequently, cells switch from the tricarboxylic acid (TCA) cycle respiration to glycolysis, reducing sensitivity to the copper ionophore elesclomol (ES). Restoring STEAP4 expression or pharmacological activation of SIRT3 with honokiol (HKL) re-instated mitochondrial STEAP4 localization and re-sensitized HBV-related HCC cells to cuproptosis; combination with ES produced synergistic tumor suppression in vitro and in orthotopic models. Collectively, our findings establish the SIRT3-STEAP4 axis as a novel regulator of cuproptosis resistance in HBV-related HCC. HBx-mediated repression of SIRT3 disrupts STEAP4 deacetylation and mitochondrial targeting, fostering metabolic reprogramming and evasion of copper-induced cell death. The results provide a pre-clinical rationale for copper-directed combination strategies in HBV-associated HCC.

Severe liver injury can result from an overdose of acetaminophen (APAP). Rhein (RA) has shown a strong capability of combating inflammation and oxidative stress. Our study aimed to investigate the potential of RA in protecting the liver from APAP-induced damage and elucidate the mechanism of action. C57BL/6 mice received a pretreatment with intragastric 100 mg/kg/day RA, followed by 400 mg/kg/day APAP intraperitoneally. Liver injury was assessed via histopathology (hematoxylin and eosin (HE)-staining), serum biochemical indicators. AML-12 cells were treated with APAP or RA, and oxidative stress and apoptosis markers were evaluated. The underlying molecular mechanisms were examined by Western blotting (WB) and quantitative Real-time polymerase chain reaction (qPCR). RA treatment significantly alleviated hepatic injury caused by APAP in rodent model, reduced oxidative stress and apoptosis, and exerted a cytoprotective effect on AML-12 cells. RA increased intracellular glutathione (GSH) content and the GSH/GSSG (Glutathione disulfide) ratio, thereby enhancing the GSH redox cycle through the Nrf2 (Nuclear factor erythroid 2-related factor 2)-GCL (glutamate cysteine ligase)/GSR (glutathione-disulfide reductase) pathway. In addition, RA inhibited the expression of CYP2E1 and CYP1A2, the enzymes converting APAP into the NAPQI (N-acetyl-p-benzoquinone imine), thus effectively mitigating APAP-induced DILI (Drug Induced Liver Injury). We identified that RA prevents APAP-induced DILI liver injury by upregulating Nrf2 and inhibiting APAP-activating enzymes.

Nanoplastics (NPs) exhibit neurotoxicity, yet the precise molecular mechanisms remain elusive. In this study, we established a human-relevant polystyrene nanoplastics (PS-NPs, 50 mg kg-1) oral exposure model in C57BL/6 mice in vivo and a neuro-immune microglial-neuron co-culture system (HMC-3/SH-SY5Y cells) in vitro to dissect these mechanisms. We demonstrate that PS-NPs exposure triggers microglial M1 activation and drives neuronal senescence. Mechanistically, PS-NPs activate the protein phosphatase 2A (PP2A)-B56γ subunit, which selectively dephosphorylates the ribosome biogenesis regulator ErbB3-binding protein 1 (Ebp1) at Ser335. This post-translational modification reduces Ebp1 nucleolar localization, suppresses 47S pre-ribosomal RNA transcription, and induces nucleolar stress. Consequently, the p53/p21 pathway is engaged, promoting neuronal senescence. Pharmacological inhibition of PP2A with LB-100 restored ribosome biogenesis, prevented neuronal senescence, and rescued cognitive deficits and neurodegenerative phenotypes in PS-NP-exposed mice. This is the first study to identify the PP2A-B56γ-p-Ebp1Ser335-ribosome biogenesis axis as a novel cascade mechanism driving PS-NP-induced neuronal senescence. Our findings offer a targetable strategy to mitigate nanoplastics-associated neurodegeneration.

Optical coherence tomography angiography (OCTA) is a major advancement in imaging, offering high-resolution microvascular volumetric images crucial for diagnosing and studying dermatological diseases. However, current data analysis and clinical evaluation criteria primarily rely on 2-dimensional (2D) imaging results, resulting in imprecise diagnoses due to the substantial loss of 3D curved structures and microvascular details. To address this issue, we propose a high-fidelity 3D curved processing workflow that integrates an artificial neural network (ANN) with a 3D denoising algorithm based on the curvelet transform and optimal orientation flow (OOF). This innovative workflow enables precise 3D segmentation and accurate quantification of dermal layer microvasculature in atopic dermatitis (AD) in vivo. Furthermore, the use of 3D multiparametric microvasculature quantitative metrics establishes a robust framework for assessing the efficacy of AD treatments in 3D images. Our study results demonstrate that skin structure imaging and the dynamic evolution of 3D microvasculature align with observed pathological changes. Compared to traditional 2D analysis, the maximum variation rate of 3D curved multiparametric information is approximately 10%. Consequently, our research marks a significant advancement in the accurate quantification of microvasculature in AD development and theranostics, paving the way for the clinical application of OCTA in dermatology.

Microcystin‐LR (MC‐LR) exacerbates metabolic dysfunction‐associated steatotic liver disease (MASLD) by inducing histopathological damage and lipid metabolism disorders. Inducible hepatocyte‐derived extracellular vesicles (iHD‐EVs) released after xenobiotic exposure activate the macrophage NOD‐like receptor protein 3 (NLRP3) inflammasome. Suppression of NLRP3 phosphorylation at serine 295 (p‐NLRP3 S295 ) is previously shown to alleviate MASLD progression. Here, it is demonstrated that microcystin‐LR (MC‐LR)‐induced iHD‐EVs reduced deliver of miR‐328‐3p to macrophages, thereby upregulating protein phosphatase 2A (PP2A)‐B56δ. Consequent PP2A‐B56δ activation disrupts inositol 1,4,5‐triphosphate receptor and voltage‐dependent anion channel 1 coupling, evokes mitochondria‐associated endoplasmic reticulum membrane (MAM) calcium (Ca 2+ ) overload, and recruits p‐NLRP3 S295 into the inflammasome. Neutralization of p‐NLRP3 S295 with a site‐specific monoclonal antibody (anti‐p‐NLRP3 S295 mAb) markedly attenuates liver inflammation and injury in MC‐LR‐exposed mice. Collectively, the miR‐328‐3p/PP2A‐B56δ/p‐NLRP3 S295 axis is identified as a crucial driver of metaflammation and establishes circulating EV‐miR‐328‐3p as a novel biomarker and anti‐p‐NLRP3 S295 mAbs as translational tools for MC‐LR‐associated MASLD.

Optical coherence tomography angiography (OCTA) is a major advancement in imaging, offering high-resolution microvascular volumetric images crucial for diagnosing and studying dermatological diseases. However, current data analysis and clinical evaluation criteria primarily rely on 2-dimensional (2D) imaging results, resulting in imprecise diagnoses due to the substantial loss of 3D curved structures and microvascular details. To address this issue, we propose a high-fidelity 3D curved processing workflow that integrates an artificial neural network (ANN) with a 3D denoising algorithm based on the curvelet transform and optimal orientation flow (OOF). This innovative workflow enables precise 3D segmentation and accurate quantification of dermal layer microvasculature in atopic dermatitis (AD) in vivo. Furthermore, the use of 3D multiparametric microvasculature quantitative metrics establishes a robust framework for assessing the efficacy of AD treatments in 3D images. Our study results demonstrate that skin structure imaging and the dynamic evolution of 3D microvasculature align with observed pathological changes. Compared to traditional 2D analysis, the maximum variation rate of 3D curved multiparametric information is approximately 10%. Consequently, our research marks a significant advancement in the accurate quantification of microvasculature in AD development and theranostics, paving the way for the clinical application of OCTA in dermatology.

BACKGROUND Previous cellular studies have demonstrated that elevated expression of Cx43 promotes the degradation of cyclin E1 and inhibits cell proliferation through ubiquitination. Conversely, reduced expression results in a loss of this capacity to facilitate cyclin E degradation. The ubiquitination and degradation of cyclin E1 may be associated with phosphorylation at specific sites on the protein, with Cx43 potentially enhancing this process by facilitating the phosphorylation of these critical residues. AIM To investigate the correlation between expression of Cx43, SKP1/Cullin1/F-box (SCF)FBXW7, p-cyclin E1 (ser73, thr77, thr395) and clinicopathological indexes in colon cancer. METHODS Expression levels of Cx43, SCFFBXW7, p-cyclin E1 (ser73, thr77, thr395) in 38 clinical colon cancer samples were detected by immunohistochemistry and were analyzed by statistical methods to discuss their correlations. RESULTS Positive rate of Cx43, SCFFBXW7, p-cyclin E1(Ser73), p-cyclin E1 (Thr77) and p-cyclin E1 (Thr395) in detected samples were 76.32%, 76.32%, 65.79%, 5.26% and 55.26% respectively. Positive expressions of these proteins were not related to the tissue type, degree of tissue differentiation or lymph node metastasis. Cx43 and SCFFBXW7(r = 0.749), p-cyclin E1 (Ser73) (r = 0.667) and p-cyclin E1 (Thr395) (r = 0.457), SCFFBXW7 and p-cyclin E1 (Ser73) (r = 0.703) and p-cyclin E1 (Thr395) (0.415) were correlated in colon cancer (P < 0.05), and expressions of the above proteins were positively correlated in colon cancer. CONCLUSION Cx43 may facilitate the phosphorylation of cyclin E1 at the Ser73 and Thr195 sites through its interaction with SCFFBXW7, thereby influencing the ubiquitination and degradation of cyclin E1.

Background & AimsMechanisms behind the impaired response of antigen-specific B cells to therapeutic vaccination in chronic hepatitis B virus (HBV) infection remain unclear. The development of vaccines or strategies to overcome this obstacle is vital for advancing the management of chronic hepatitis B.MethodsA mouse model, denominated as E6F6-B, was engineered to feature a knock-in of a B-cell receptor (BCR) that specifically recognizes HBsAg. This model served as a valuable tool for investigating the temporal and spatial dynamics of humoral responses following therapeutic vaccination under continuous antigen exposure. Using a suite of immunological techniques, we elucidated the differentiation trajectory of HBsAg-specific B cells post-therapeutic vaccination in HBV carrier mice.ResultsUtilizing the E6F6-B transfer model, we observed a marked decline in antibody-secreting cells 2 weeks after vaccination. A dysfunctional and atypical pre-plasma cell population (BLIMP-1+ IRF4+ CD40- CD138- BCMA-) emerged, manifested by sustained BCR signaling. By deploying an antibody to purge persistent HBsAg, we effectively prompted the therapeutic vaccine to provoke conventional plasma cell differentiation. This resulted in an enhanced anti-HBs antibody response and facilitated HBsAg clearance.ConclusionsSustained high levels of HBsAg limit the ability of therapeutic hepatitis B vaccines to induce the canonical plasma cell differentiation necessary for anti-HBs antibody production. Employing a strategy combining antibodies with vaccines can surmount this altered humoral response associated with atypical pre-plasma cells, leading to improved therapeutic efficacy in HBV carrier mice.Impact and implicationsTherapeutic vaccines aimed at combatting HBV encounter suboptimal humoral responses in clinical settings, and the mechanisms impeding their effectiveness have remained obscure. Our research, utilizing the innovative E6F6-B mouse transfer model, reveals that the persistence of HBsAg can lead to the emergence of an atypical pre-plasma cell population, which proves to be relevant to the potency of therapeutic HBV vaccines. Targeting the aberrant differentiation process of these atypical pre-plasma cells stands out as a critical strategy to amplify the humoral response elicited by HBV therapeutic vaccines in carrier mouse models. This discovery suggests a compelling avenue for further study in the context of human chronic hepatitis B. Encouragingly, our findings indicate that synergistic therapy combining HBV-specific antibodies with vaccines offers a promising approach that could significantly advance the pursuit of a functional cure for HBV.

This study demonstrated that both copper oxide nanoparticles (CuO-NPs) and copper nanoparticles (Cu-NPs) can cause swelling, inflammation, and cause damage to the mitochondria of alveolar type II epithelial cells in mice. Cellular examinations indicated that both CuO-NPs and Cu-NPs can reduce cell viability and harm the mitochondria of human bronchial epithelial cells, particularly Beas-2B cells. However, it is clear that CuO-NPs exhibit a more pronounced detrimental effect compared with Cu-NPs. Using bafilomycin A1 (Bafi A1), an inhibitor of lysosomal acidification, was found to enhance cell viability and alleviate mitochondrial damage caused by CuO-NPs. Additionally, Bafi A1 also reduces the accumulation of dihydrolipoamide S-acetyltransferase (DLAT), a marker for mitochondrial protein toxicity, induced by CuO-NPs. This observation suggests that the toxicity of CuO-NPs depends on the distribution of copper particles within cells, a process facilitated by the acidic environment of lysosomes. The release of copper ions is thought to be triggered by the acidic conditions within lysosomes, which aligns with the lysosomal Trojan horse mechanism. However, this association does not seem to be evident with Cu-NPs.

Helical micro-/nanoswimmers have been extensively explored as a miniaturized robotic tool for advanced biomedicine in the past two decades. However, the integration of biomedical functions on the existing swimmers remains insufficient to allow further in vivo developments. We propose a chemically engineered Spirulina microalgae with biogenic gold nanoparticles deposited in its cells and magnetite nanoparticles assembled on its surface. This biohybrid helical microswimmer is naturally of many desired functions and can be easily functionalized through surface modification and/or intracellular loading. The gold-enabled digital subtraction angiography of multimodal imaging function is highlighted herein. Such imaging modality makes real-time tracking possible in deep anatomical regions, guaranteeing precise in vivo navigation of our swimmers to specific lesions. Meanwhile, the gold-enhanced photothermal efficacy is also noteworthy. It regulates swimmer degradation via controlling photothermal heat and can be harnessed to implement targeted therapy, for which the targeting process and therapeutic effect are successfully demonstrated in a fluidic chip. Furthermore, the fabrication method is straightforward and cost-effective, suitable for mass production. All these results clarify the functionalization strategy enabled by biogenic intracellular deposition and provide a multifunctional biohybrid microswimmer.

A-kinase anchoring protein 95 (AKAP95) functions as a scaffold for protein kinase A. Prior work by our group has shown that AKAP95, in coordination with Connexin 43 (Cx43), modulates the expression of cyclin D and E proteins, thus affecting the cell cycle progression in lung cancer cells. In the current study, we confirmed that AKAP95 forms a complex with Cx43. Moreover, it associates with cyclins D1 and E1 during the G1 phase, leading to the formation of protein complexes that subsequently translocate to the nucleus. These findings indicate that AKAP95 might facilitate the nuclear transport of cyclins D1 and E1. Throughout this process, AKAP95 and Cx43 collectively regulate the expression of cyclin D, phosphorylate cyclin E1 proteins, and target their specific ubiquitin ligases, ultimately impacting cell cycle progression.

Cerebral ischemia-reperfusion injury (CIRI) is a major challenge to neuronal survival in acute ischemic stroke (AIS). However, effective neuroprotective agents remain to be developed for the treatment of CIRI. In this work, we have developed an Anti-TRAIL protein-modified and indocyanine green (ICG)-responsive nanoagent (Anti-TRAIL-ICG) to target ischemic areas and then reduce CIRI and rescue the ischemic penumbra. In vitro and in vivo experiments have demonstrated that the carrier-free nanoagent can enhance drug transport across the blood-brain barrier (BBB) in stroke mice, exhibiting high targeting ability and good biocompatibility. Anti-TRAIL-ICG nanoagent played a better neuroprotective role by reducing apoptosis and ferroptosis, and significantly improved ischemia-reperfusion injury. Moreover, the multimodal imaging platform enables the dynamic in vivo examination of multiple morphofunctional information, so that the dynamic molecular events of nanoagent can be detected continuously and in real time for early treatment in transient middle cerebral artery occlusion (tMCAO) models. Furthermore, it has been found that Anti-TRAIL-ICG has great potential in the functional reconstruction of neurovascular networks through optical coherence tomography angiography (OCTA). Taken together, our work effectively alleviates CIRI after stoke by blocking multiple cell death pathways, which offers an innovative strategy for harnessing the apoptosis and ferroptosis against CIRI.

Micro/Nano-scale particles are widely used as vaccine adjuvants to enhance immune response and improve antigen stability. While aluminum salt is one of the most common adjuvants approved for human use, its immunostimulatory capacity is suboptimal. In this study, we modified risedronate, an immunostimulant and anti-osteoporotic drug, to create zinc salt particle-based risedronate (Zn-RS), also termed particulate risedronate. Compared to soluble risedronate, micronanoparticled Zn-RS adjuvant demonstrated increased recruitment of innate cells, enhanced antigen uptake locally, and a similar antigen depot effect as aluminum salt. Furthermore, Zn-RS adjuvant directly and quickly stimulated immune cells, accelerated the formulation of germinal centers in lymph nodes, and facilitated the rapid production of antibodies. Importantly, Zn-RS adjuvant exhibited superior performance in both young and aged mice, effectively protecting against respiratory diseases such as SARS-CoV-2 challenge. Consequently, particulate risedronate showed great potential as an immune-enhancing vaccine adjuvant, particularly beneficial for vaccines targeting the susceptible elderly.

Black Phosphorus Quantum Dots (BP-QDs) have potential applications in biomedicine. BP-QDs may enter the body through the respiratory tract during grinding and crushing production and processing, causing respiratory toxicity. Ferroptosis is an oxidative, iron-dependent form of cell death. Here, respiratory toxicity of BP-QDs has been validated in mice and human bronchial epithelial cells. After 24 h of exposure to different doses (4-32 & mu;g/ mL) of BP-QDs, intracellular lipid peroxidation and iron overload occurred in Beas-2B cells. After 4 times exposures by noninvasive tracheal instillation at four doses [0, 0.25, 0.5 and 1 (mg/kg/48h)], all animals were sacrificed, organs were removed, processed for pathological examination and molecular analysis. Iron overload, glutathione (GSH) depletion and lipid peroxidation in the lung tissue of mice in the exposure group. Furthermore, based on the ferroptosis-associated protein and mRNA expression, it was hypothesized that BP-QDs induced ferroptosis through increasing intracellular free iron and polyunsaturated fatty acid synthesis. By comparing with previous studies, we speculate that primary cells generally are more sensitive to BP-QDs-induced damage than cancer cells. In summary, findings in the present study confirmed that BP-QDs induce ferroptosis via increasing lipid peroxidation and iron accumulation in vitro and in vivo.

Periodontitis, one of the most common dental diseases, is the main cause of tooth loss. Bacterial infection and the host immune response are the dominant factors that determine periodontal disease. Therefore, the key factors in the treatment of periodontitis are to remove pathogenic factors and repair periodontal supporting tissues. In this work, a facile and versatile strategy to prepare pH-sensitive biomineralized nanoparticles constructed with antibiotic to simultaneously deliver antibacterial, anti-inflammatory, and bone repair capabilities is presented. The experiments in vitro show that the nanoreactor achieves good inhibitory effects on three common periodontal pathogens and has activity against damaging inflammatory mediators, reducing lipopolysaccharide-induced inflammatory responses. In vivo CT images of the maxillary molar area and histomorphological and immunohistochemical analyses demonstrate that the nanoparticles repair function for periodontal supporting tissues, reverse the loosened alveolar bone tissue, and remodel the periodontal inflammatory microenvironment. Therefore, the present research provides a feasible approach to both prevent the loss of alveolar bone and repair periodontal tissue.

PURPOSE:The board application of black phosphorus quantum dots (BP-QDs) increases the risk of inhalation exposure in the manufacturing process. The aim of this study is to explore the toxic effect of BP-QDs on human bronchial epithelial cells (Beas-2B) and lung tissue of Balb/c mice.METHODS:The BP-QDs were characterized using transmission electron microscopy (TEM) and a Malvern laser particle size analyzer. Cell Counting Kit-8 (CCK-8) and TEM were used to detect cytotoxicity and organelle injury. Damage to the endoplasmic reticulum (ER) was detected by using the ER-Tracker molecular probe. Rates of apoptosis were detected by AnnexinV/PI staining. Phagocytic acid vesicles were detected using AO staining. Western blotting and immunohistochemistry were used to examine the molecular mechanisms.RESULTS:After treatment with different concentrations of BP-QDs for 24 h, the cell viability decreased, as well as activation of the ER stress and autophagy. Furthermore, the rate of apoptosis was increased. Inhibition of ER stress caused by 4-phenyl butyric acid (4-PBA) was shown to significantly inhibit both apoptosis and autophagy, suggesting that ER stress could be an upstream mediator of both autophagy and apoptosis. BP-QD-induced autophagy can also inhibit the occurrence of apoptosis using molecules related to autophagy including rapamycin (Rapa), 3-methyladenine (3-MA), and bafilomycin A1 (Bafi A1). In general, BP-QDs activate ER stress in Beas-2B cells, which further induces autophagy and apoptosis, and autophagy may be activated as a factor that protects against apoptosis. We also observed strong staining of related proteins of ER stress, autophagy, and apoptosis proteins in mouse lung tissue following intracheal instillation over the course of a week.CONCLUSION:BP-QD-induced ER stress facilitates autophagy and apoptosis in Beas-2B cells and autophagy may be activated as a protective factor against apoptosis. Under conditions of ER stress induced by BP-QDs, The interplay between autophagy and apoptosis determines cell fate.

Sonodynamic bacterial inactivation, a reactive oxygen species (ROS)-empowered approach featuring high penetration depth and low health risk, is explored for antibiotic-free antibacterial treatment. However, the low yield and insufficient diffusion of ROS negatively affect the antibacterial efficacy of sonodynamic treatment, thus hindering its further development. Here an actuator-integrated mechanism is proposed for enhancing the sonodynamic efficacy of loaded sonosensitizers through motion-induced hydrodynamic effects, demonstrated by a porphyrin-decorated gold nanomotor, which can produce ROS for bacterial inactivation while performing multimodal motion via actuation using low-frequency ultrasound. Corroborated by numerical simulation, the experimental results show that the motor's stirring motion significantly increases the yield and diffusion of ROS through fluid flow and frequent interactions between the motor and bacterial targets, resulting in doubled antibacterial efficiency in comparison to a stationary motor. Furthermore, the flow-induced shear forces combined with the frequent interactions constitute a source of mechanical damage and can form a synergy with the antibacterial properties of ROS, enabling an efficient biofilm eradication that is inaccessible by freely suspended porphyrin. In conclusion, this study reports a motion-based strategy to enhance sonodynamic efficacy and provides proof of concept using a sonodynamic gold nanomotor powered by ultrasound.

To investigate the effect of resonant thermal radiation from electric graphene films (GFs) to the immunoactivity of T cells in situ and the benefits to cancer immunotherapy. This work utilizes externally applied GFs to regulate the immunoactivity of T cells through the eminent resonant thermal radiation effect. The GFs are fabricated with high emissivity and well‐matched far‐infrared emission peaks between the GFs and living bodies. The T‐cell antigen receptor signaling mechanisms and the ensuing immunoactivity of T cells by GFs treatment, compared with traditional metal films (MFs), are studied both in vitro and in vivo. The resonance thermal radiation decreases the activation threshold of T cells by increasing the influx of calcium ions, neutralizing the negative charge of intracellular membranes and triggering the phospho‐CD3ζ event. As a result, GF treatment elicits an enhanced T‐cell‐based immune response and serves as an externally applied “adjuvant” to T‐cell activation, which shows remarkable efficiency in combination with aPD‐L1 immunotherapy in multiple mouse models. This work centers the unmet needs of regulating the immunoactivity of intratumoral T cells in situ, in which the auxiliary use of GFs with high efficiency and little safety concern has great promise to translational study regarding cancer immunotherapy in future.

Optical coherence tomography angiography (OCTA) images suffer from inevitable micromotion (breathing, heartbeat, and blinking) noise. These image artifacts can severely disturb the visibility of results and reduce accuracy of vessel morphological and functional metrics quantization. Herein, we propose a multiple wavelet-FFT algorithm (MW-FFTA) comprising multiple integrated processes combined with wavelet-FFT and minimum reconstruction that can be used to effectively attenuate motion artifacts and significantly improve the precision of quantitative information. We verified the fidelity of image information and reliability of MW-FFTA by the image quality evaluation. The efficiency and robustness of MW-FFTA was validated by the vessel parameters on multi-scene in vivo OCTA imaging. Compared with previous algorithms, our method provides better visual and quantitative results. Therefore, the MW-FFTA possesses the potential capacity to improve the diagnosis of clinical diseases with OCTA.

Surgical resection (SR) is the major method for treating invasive or recurrent noninvasive bladder cancer (BC). However, SR still causes additional risks of urinary tract infection, bleeding and some complications that may be life-threatening. Therefore, it is of urgent medical need for new diagnosis and therapy scheme for BC. Herein, we proposed a strategy that combines dual-modality three-dimensional (3D) NIR-II photoacoustic (PA) and optical coherence tomography angiography (OCTA) molecular imaging to guide visually interventional photothermal therapy (PTT) for BC. Additionally, we developed an excellent NIR-II hyaluronic acid-IR-1048 with liposome- coated (HAPO-1048) photothermal agent for PTT of CD44 overexpressing orthotopic BC, which showed strong NIR-II optical absorption, preferable tumor targeting, excellent biocompatibility and high PTT efficacy. Combined with the NIR-II PA/PTT response of the HAPO-1048, we can achieve high sensitivity and specificity (about 2 mm diameter) dynamic monitoring and investigate the precision therapy of PTT in deep orthotopic BC (4.38 mm). Simultaneously, we can map the dynamic changes of 3D PA signals and HAPO-1048 nanoparticles (HAPO-1048 NPs) enrichment, which is closely related to the PTT efficacy. Significantly, the whole changes of complex microvascular morphometrics and angiogenesis of BC were well surveilled and assessed in vivo using OCTA. Lastly, minimally invasive interventional PTT strategy was successfully implemented to achieve a controllable and precise therapy for orthotopic BC based on laparoscopy and US guidance. In conclusion, we present a multimodal NIR-II optical imaging synergy laparoscopy interventional oncologic scheme, which should expand the current PTT technology, leading to a significant ongoing in treatment of primary tumors of urinary system