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Mechanisms 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. A 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. Utilizing 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. Sustained 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. Therapeutic 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. [Display omitted] • The HBsAg-specific BCR knock-in mouse was constructed to study the characteristics of B cells in chronic HBV infection. • An atypical pre-plasma cell population was identified post therapeutic vaccination in chronic HBV carrier mice. • Atypical pre-plasma cells display enhanced BCR signaling, ER stress and apoptosis-related gene expression. • Combination therapy (antibody + vaccine) partially overcame abnormal B-cell differentiation and led to superior HBsAg suppression. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

• Chemically-engineered Spirulina as a multifunctional swimming microrobot. • Biogenic intracellular deposition of gold nanoparticles. • Gold-enhanced imaging and photothermal efficacy. • Digital subtraction angiography enabled real-time yet deep tracking. 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. [ABSTRACT FROM AUTHOR]

Background: This study aimed to overexpress or silence connexin 43 (Cx43) and A‐kinase anchoring protein 95 (AKAP95) in human A549 cells to explore their effects on cyclins and on G1/S conversion when the interrelationship of Cx43, AKAP95, and cyclin E1/E2 changes. Methods: The study mainly used Western blot analysis and Co‐immuno precipitation to detect the target protein in Cx43/AKAP95 over expressed human A549 cells, and the relationship of proteins Cx43, AKAP95 and Cyclin E during G1‐S phase was explored with qualitative and quantitative analysis. Results: The overexpression of Cx43 inhibited the expression of cyclin D1 and E1 by accelerating their degradation and reduced the Cdk2 activity that blocked the DNA transcription activity. However, the overexpression of AKAP95 increased the expression of cyclin D1 and E1 and inhibited their degradation, and enhanced the Cdk2 activity that promoted the DNA transcription activity. Cx43 and AKAP95 competitively bound to cyclin E1/E2, and the competitive binding affected the Cdk2 activity, Rb phosphorylation, DNA transcription activity, and G1/S conversion. Conclusions: This study showed that the expression of ERK1/2, PKA, and PKB increased when BEAS‐2B cells were treated with PDGF‐BB, suggesting that ERK1/2, PKA, and PKB might be involved in the binding of AKAP95 with cyclin E, or the separation of AKAP95 from Cx43 from cyclin E1/E2. The specific mechanism underlying this process still needs further exploration. [ABSTRACT FROM AUTHOR]

Aim: This study aimed to investigate the correlation between the expression of A‐kinase anchor protein95 (AKAP95), p‐retinoblastoma (phosphorylated Rb, p‐Rb), cyclin D2, cyclin D3 and cyclin E2 in esophageal cancer tissues and clinicopathological indexes. Method: The protein expression levels of AKAP95, p‐Rb, cyclin D2/3 and cyclin E2 in 40 esophageal cancer tissues were detected using immunohistochemistry, and the correlation between them was analyzed. Result: The percentage of p‐Rb (Ser780)‐, cyclin D2‐, cyclin D3‐ and cyclin E2‐positive samples was 62.50%, 70.00%, 67.50% and 60.00%, respectively. Also, the positive expression did not correlate with the histological type, histological differentiation or lymph node metastasis. The expression of AKAP95 and p‐Rb (Ser780), p‐Rb (Ser780) and cyclin D2 and p‐Rb (Ser780) and cyclin D3 in esophageal cancer tissues was found to be correlated (P < 0.05). Conclusions: The expression of AKAP95 and p‐Rb (Ser780), p‐Rb(Ser780) and cyclin D2, and p‐Rb (Ser780) and cyclin D3 in esophageal cancer tissue was correlated, suggesting that these proteins might play a synergistic role in cell‐cycle progression. Cyclin D2/D3 and p‐Rb (Ser780) were correlated whereas cyclin E2 and p‐Rb (Ser780) were not, suggesting that p‐Rb (Ser780) might be highly expressed and the Ser780 site of Rb protein might be phosphorylated in the early stage of the G1 phase. Ser780 was the site in the primary phosphorylation stage of several phosphorylation sites during stepwise phosphorylation (from primary to high phosphorylation). [ABSTRACT FROM AUTHOR]

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 μg/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. [Display omitted] • BP-QDs entered by the respiratory tract could cause lung injury. • BP-QDs induced ferroptosis in vitro and in vivo. • BP-QDs might induce ferroptosis by altering intracellular iron homeostasis. • BP-QDs might promote the synthesis of polyunsaturated fatty acids in cells. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • Repeated infusion of BP-QDs for a week resulted in lung injury to mice. • BP-QD-induced ER stress facilitates autophagy and apoptosis in Beas-2B cells. • 4-PBA can partly resist autophagy and apoptosis induced by BP-QDs. • Defective autophagy or dysfunction of autophagic flux promotes cell apoptosis. • Autophagy may be activated as a protective factor against apoptosis. [ABSTRACT FROM AUTHOR]

Background This study was conducted to investigate the exchange protein directly activated by c AMP ( Epac1), PDE4, and PKC expression in breast cancer tissues, and the correlation between these proteins and AKAP95, Cx43, cyclin D2, and cyclin E1. Methods PV-9000 two-step immunohistochemistry was used to analyze protein expression. Results The positive rate of Epac1 protein expression in breast cancer tissues (58%) was higher than in para-carcinoma tissues (10%) ( P < 0.05). There were no significant differences in the positive rates of PDE4 and PKC expression between breast cancer and para-carcinoma tissues ( P > 0.05). The positive expression rate of PDE4 was higher in the P53 protein positive group compared to the P53 negative group ( P < 0.05). Correlations between Epac1 and cyclin D2, PDE4 and cyclin D2, AKAP95 and PKC, Cx43 and PKC, and cyclin D2 and PKC proteins were observed ( P < 0.05). Conclusion Epac1 expression in breast cancer tissues was increased, suggesting that the protein may be involved in the development of breast cancer. Correlations between Epac1 and cyclin D2, PDE4 and cyclin D2, AKAP95 and PKC, Cx43 and PKC, and cyclin D2 and PKC proteins suggested synergistic effects among these proteins in the development of breast cancer. [ABSTRACT FROM AUTHOR]

• An excellent NIR-II PTA (HAPO-1048) for actively-targeted PTT of orthotopic BC. • Combining dual-modality 3D NIR-II PA and OCTA molecular imaging for micro-BC. • High sensitivity and specificity (about 2 mm diameter) dynamic detecting of BC. • Designing a minimally invasive interventional PTT for deep orthotopic BC. • HAPO-1048 demonstrates improved NIR-II therapeutic efficiency. 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. [ABSTRACT FROM AUTHOR]

Arsenic is a widely existing pollutant in the environment, but the mechanism of occurrence and development of lung cancer by long-term arsenic exposure needs to be elucidated further. How the high and low doses of arsenic induce human bronchial epithelial cell transformation is yet to be elucidated. In the present study, human bronchial epithelial cells were exposed to varying high-dose sodium arsenite (NaAsO 2) for the short-term or treated with low dose for long-term. The data showed that both short- and long-term treatment promoted G1/S transition of Beas-2B cells, inducing a significant increase in the expression of AKAP95, cyclin D1, cyclin D2, and cyclin E1. However, silencing AKAP95 by treating cells with siAKAP95 exerted a protective function that inhibited G1/S transition, suggesting a regulatory mechanism of AKAP95 on the cell cycle during cell malignant transformation induced by NaAsO 2. In addition, mitochondrial dysfunctions occurred during NaAsO 2 exposure. Beas-2B cells exposed to low-dose NaAsO 2 for long-term were subcultured for 20 generations, and the exposure time was positively proportional to the growth and migration rate of the cells. The exposed cells were used in a tumor-bearing transplantation experiment (mice), and the results showed that the longer the exposure time, the faster the tumor volume growth rate of As-Beas-2B cells. Tumor tissues were excised for hematoxylin-eosin staining, which showed altered cell morphology and increased volume. [Display omitted] • Exposure to arsenic altered the cell cycle of human bronchial epithelial cells. • AKAP95 mediates cell cycle alterations induced by arsenic. • Arsenic exposure causes mitochondrial dysfunction and damage. [ABSTRACT FROM AUTHOR]

Background This study examined the expression of exchange protein directly activated by c AMP1 ( Epac1), PDE4, and PKC in esophageal cancer tissues, and analyzed the association of each protein with the pathological parameters of the samples. Methods Epac1, PDE4, and PKC protein expression was evaluated by PV-9000 two-step immunohistochemical techniques in 51 esophageal cancer specimens and 10 para-carcinoma tissues. Results The positive expression rates of Epac1 and PKC in esophageal cancer tissues (62.7% and 68.6%, respectively) were higher compared to those in para-carcinoma tissues (20% and 20%, respectively) ( P < 0.05). The positive expression rate of PDE4 in esophageal cancer tissues (54.1%) was higher than in para-carcinoma tissues (30%), ( P > 0.05). Epac1, PDE4, and PKC protein expression levels were not associated with the extent of tumor differentiation and/or lymph node metastasis ( P > 0.05). Epac1 protein expression levels correlated with PDE4, PKC, and AKAP95 protein expression levels. In addition, there was a correlation between PKC and Cx43 protein levels ( P < 0.05). Conclusion The expression rates of Epac1, PDE4, and PKC protein in esophageal cancer tissues were significantly higher compared to the rates in para-carcinoma tissues, suggesting an association between these proteins and the development and progression of esophageal cancer. The correlations between these proteins also revealed that they may exert a synergistic effect during the development of esophageal cancer. [ABSTRACT FROM AUTHOR]