AIEgens Cross-linked Iron Oxide Nanoparticles Synchronously Amplify Bimodal Imaging Signals in Situ by Tumor Acidity-Mediated Click Reaction.

Activatable dual-modal molecular imaging probes present a promising tool for the diagnosis of malignant tumors. However, synchronously enhancing dual-modal imaging signals under a single stimulus is challenging. Herein, we propose an activatable bimodal probe that integrates aggregation-induced emission luminogens (AIEgens) and iron oxide nanoparticles (IOs) to synergistically enhance near-infrared fluorescence (NIRF) intensity and magnetic resonance (MR) contrast through a tumor acidity-mediated click reaction. Tumor acidity-responsive IOs containing dibenzocyclooctyne groups (termed cDIOs) and AIEgens containing azide groups (termed AATs) can be covalently cross-linked in response to tumor acidity, which leads to a simultaneous enhancement in NIRF intensity (≈12.4-fold) and r2 relaxivity (≈2.8-fold). cDIOs and AATs were effectively activated in mice orthotropic breast tumor, and the cross-linking prolonged their retention in tumor, further augmenting the bimodal signals and expanding imaging time frame. This facile strategy leverages the inherent properties of probes themselves and demonstrates promise in future translational studies.

Prediction of 30-day mortality in heart failure patients with hypoxic hepatitis: Development and external validation of an interpretable machine learning model.

Background: This study aimed to explore the impact of hypoxic hepatitis (HH) on survival in heart failure (HF) patients and to develop an effective machine learning model to predict 30-day mortality risk in HF patients with HH. Methods: In the Medical Information Mart for Intensive Care (MIMIC)-III and IV databases, clinical data and survival situations of HF patients admitted to the intensive care unit (ICU) were retrospectively collected. Propensity Score Matching (PSM) analysis was used to balance baseline differences between HF patients with and without HH. Kaplan Meier analysis and multivariate Cox analysis were used to determining the effect of HH on the survival of CF patients. For developing a model that can predict 30-day mortality in CF patients with HH, the feature recurrence elimination (RFE) method was applied to feature selection, and seven machine learning algorithms were employed to model construction. After training and hyper-parameter optimization (HPO) of the model through cross-validation in the training set, a performance comparison was performed through internal and external validation. To interpret the optimal model, Shapley Additive Explanations (SHAP) were used along with the Local Interpretable Model-agnostic Explanations (LIME) and the Partial Dependence Plot (PDP) techniques. Results: The incidence of HH was 6.5% in HF patients in the MIMIC cohort. HF patients with HH had a 30-day mortality rate of 33% and a 1-year mortality rate of 51%, and HH was an independent risk factor for increased short-term and long-term mortality risk in HF patients. After RFE, 21 key features (21/56) were selected to build the model. Internal validation and external validation suggested that Categorical Boosting (Catboost) had a higher discriminatory capability than the other models (internal validation: AUC, 0.832; 95% CI, 0.819-0.845; external validation: AUC, 0.757 95% CI, 0.739-0.776), and the simplified Catboost model (S-Catboost) also had good performance in both internal validation and external validation (internal validation: AUC, 0.801; 95% CI, 0.787-0.813; external validation: AUC, 0.729, 95% CI, 0.711-0.745). Conclusion: HH was associated with increased mortality in HF patients. Machine learning methods had good performance in identifying the 30-day mortality risk of HF with HH. With interpretability techniques, the transparency of machine learning models has been enhanced to facilitate user understanding of the prediction results.

Cinnamaldehyde-based poly(thioacetal): A ROS-awakened self-amplifying degradable polymer for enhanced cancer immunotherapy.

Although stimuli-responsive polymers have emerged as promising strategies for intelligent cancer therapy, limited polymer degradation and insufficient drug release remain a challenge. Here, we report a novel reactive oxygen species (ROS)-awakened self-amplifying degradable cinnamaldehyde (CA)-based poly(thioacetal) polymer. The polymer consists of ROS responsive thioacetal (TA) group and CA as the ROS generation agent. The self-amplified polymer degradation process is triggered by endogenous ROS-induced cleavage of the TA group to release CA. The CA released then promotes the generation of more ROS through mitochondrial dysfunction, resulting in amplified polymer degradation. More importantly, poly(thioacetal) itself can trigger immunogenic cell death (ICD) of the tumor cells and its side chains can be conjugated with indoleamine 2,3-dioxygenase 1 (IDO-1) inhibitor to reverse the immunosuppressive tumor microenvironment for synergistic cancer immunotherapy. The self-amplified degradable poly(thioacetal) developed in this work provides insights into the development of novel stimulus-responsive polymers for enhanced cancer immunotherapy.

Spatial specific delivery of combinational chemotherapeutics to combat intratumoral heterogeneity.

Hypoxia-induced intratumoral heterogeneity poses a major challenge in tumor therapy due to the varying susceptibility to chemotherapy. Moreover, the spatial distribution patterns of hypoxic and normoxic tissues makes conventional combination therapy less effective. In this study, a tumor-acidity and bioorthogonal chemistry mediated in situ size transformable nanocarrier (NP@DOXDBCO plus iCPPAN3) was developed to spatially deliver two combinational chemotherapeutic drugs (doxorubicin (DOX) and PR104A) to combat hypoxia-induced intratumoral heterogeneity. DOX is highly toxic to tumor cells in normoxia state but less toxic in hypoxia state due to the hypoxia-induced chemoresistance. Meanwhile, PR104A is a hypoxia-activated prodrug has less toxic in normoxia state. Two nanocarriers, NP@DOXDBCO and iCPPAN3, can cross-link near the blood vessel extravasation sites through tumor acidity responsive bioorthogonal click chemistry to enhance the retention of DOX in tumor normoxia. Moreover, PR104A conjugated to the small-sized dendritic polyamidoamine (PAMAM) released under tumor acidity can penetrate deep tumor tissues for hypoxic tumor cell killing. Our study has demonstrated that this site-specific combination chemotherapy is better than the traditional combination chemotherapy. Therefore, spatial specific delivery of combinational therapeutics via in situ size transformable nanocarrier addresses the challenges of hypoxia induced intratumoral heterogeneity and provides insights into the combination therapy.

Bioorthogonal Equipping CAR-T Cells with Hyaluronidase and Checkpoint Blocking Antibody for Enhanced Solid Tumor Immunotherapy.

Adoptive cellular therapy utilizing chimeric antigen receptor redirected T (CAR-T) cells has shown impressive therapeutic effects on hematological malignancies. In contrast, the efficacy of CAR-T therapies in treating solid tumors is still poor, which is largely due to inefficient penetration into solid tumors and the immunosuppressive tumor microenvironment. Herein, we engineered hyaluronidase (HAase) and the checkpoint blocking antibody α-PDL1 on the CAR-T cell surface via highly efficient and biocompatible bioorthogonal click chemistry to improve their therapeutic effects on solid tumors. The modified HAase degrades hyaluronic acid and destroys the tumor extracellular matrix, allowing CAR-T cells to penetrate deeply into solid tumors, as evidenced by in vitro infiltration experiments and in vivo biodistribution studies. In addition, in vitro cytotoxicity studies showed stronger antitumor activity of α-PDL1-decorated cells than traditional CAR-T cells. Importantly, HAase- and α-PDL1-engineered CAR-T cells showed better therapeutic efficacy on two solid tumor models and did not cause significant systemic side effects. In this work, we provide a simple, efficient, and biologically safe chemical strategy to engineer traditional CAR-T cells for enhanced therapeutic efficacy on solid tumors, which can be extended to other adoptive cellular immunotherapies and holds great potential for clinical application.

Bioorthogonal chemistry and illumination controlled programmed size-changeable nanomedicine for synergistic photodynamic and hypoxia-activated therapy.

Photodynamic therapy (PDT) can aggravate the hypoxia aggravation and be further utilized for the activation of hypoxia-activated prodrug (HAP). Ideally, photosensitizers (PSs) are mainly administrated to tumor vasculatures adjacent to regions with high oxygen to effectively generate reactive oxygen species (ROS) effectively and further aggravate tumor hypoxia, while the HAP is delivered to the inner tumor as far as possible for efficient activation. However, a delivery system capable of transporting PSs and HAP to the desired region respectively for the optimum effect is urgently needed. Here, we developed a bioorthogonal click chemistry and illumination controlled programmed size-changeable nanomedicine for synergistic photodynamic and hypoxia-activated therapy. It utilized tumor acidity responsive bioorthogonal click reaction for crosslinking nanoparticles to construct a drug depot with tumor vasculatures adjacent region retention for PDT in normoxia. Under laser illumination, cleavage of the ROS-responsive thioketal (TK) crosslinker to release small sized poly(amidoamine) (PAMAM) dendrimer conjugated with HAP for enhanced tumor penetration into the hypoxic region. Therefore, this strategy could differentially deliver PSs and HAP in desired spatial distribution, eventually achieving the enhanced synergistic enhancement in the combined PDT and hypoxia-activated therapy.

The expression of STEAP4 in peripheral blood predicts the outcome of septic patients.

BACKGROUND: Sepsis is a systemic disease characterized by extensive inflammatory responses and impaired organ function, which are characteristics that make it easily missed and complex to treat. A large number of laboratory and clinical studies on the diagnosis and treatment of sepsis have been continuously carried out, confirming the importance of mitochondrial function during the development of sepsis. STEAP4 is an important metalloreductase in mitochondria, which is involved in the biogenesis and respiratory chain of mitochondria. The role of STEAP4 in inflammation remains controversial. Research in this field may contribute to the development of new diagnostic and treatment options for sepsis. METHODS: The expression of STEAP4 was measured in the peripheral blood of patients with severe sepsis and compared with healthy controls. Cell and mouse inflammatory models were established to detect the expression of STEAP4 and other inflammatory cytokines. RESULTS: (I) The expression of STEAP4 in the peripheral blood of patients with severe sepsis is higher than that of healthy volunteers (P<0.01), which is related to the SOFA score and transaminase. (II) STEAP4 has a certain predictive effect on the outcome of patients [area under curve (AUC) =0.696, P<0.05, 95% CI: 0.528 to 0.833]. (III) Inflammation led to increased expression of STEAP4 gene in RAW264.7 cells and mouse liver tissue. CONCLUSIONS: The expression of STEAP4 is elevated in the early stage of sepsis and the degree of its elevation can be used to predict the clinical outcome of sepsis patients.

G-rich sequence factor 1 serves as a prognostic biomarker in septic patients.

BACKGROUND: Sepsis is a condition of organ dysfunction caused by infection, and is unavoidably related to costs and mortality; however, no biomarker has yet been identified to clearly predict the prognosis of septic patients. In this study, we aimed to explore the role of guanine-rich sequence factor 1 (GRSF1) in evaluating the severity and prognosis of sepsis. METHODS: The expression of GRSF1 in peripheral blood was measured and analyzed in 42 septic participants and 32 healthy controls respectively by using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Clinical data were assessed by correlation analysis. In addition, GRSF1 expression was investigated in cecal ligation and puncture (CLP) induced mice septic models by RT-qPCR and western blot (WB). RESULTS: The expression of GRSF1 expression in septic patients in the first day of electronic intensive care unit (eICU) administration was significantly lower in comparison with HC. Further analysis showed GRSF1 expression was strongly related to the Acute Physiologic Assessment and Chronic Health Evaluation II (APACHE II) score and Sequential Organ Failure Assessment (SOFA) score. Low expression of GRSF1 predicted high mortality within 24 hours in septic patients and in CLP-induced mice. CONCLUSIONS: Decreased expression of GRSF1 was significantly correlated with high mortality in septic patients, and also in experimental septic mice. The GRSF1 protein may be a potential prognostic biomarker in sepsis.

Bioorthogonal Pretargeting Strategy for Anchoring Activatable Photosensitizers on Plasma Membranes for Effective Photodynamic Therapy.

Developing novel activatable photosensitizers with excellent plasma membrane targeting ability is urgently needed for smart photodynamic therapy (PDT). Herein, a tumor acidity-activatable photosensitizer combined with a two-step bioorthogonal pretargeting strategy to anchor photosensitizers on the plasma membrane for effective PDT is developed. Briefly, artificial receptors are first anchored on the cell plasma membrane using cell-labeling agents (Az-NPs) via the enhanced permeability and retention effect to achieve the tumor cell labeling. Then, pH-sensitive nanoparticles (S-NPs) modified with dibenzocyclooctyne (DBCO) and chlorin e6 (Ce6) accumulate in tumor tissue and disassemble upon protonation of their tertiary amines in response to the acidic tumor environment, exposing the contained DBCO and Ce6. The selective, highly specific click reactions between DBCO and azide groups enable Ce6 to be anchored on the tumor cell surface. Upon laser irradiation, the cell membrane is severely damaged by the cytotoxic reactive oxygen species, resulting in remarkable cellular apoptosis. Taken together, the membrane-localized PDT by our bioorthogonal pretargeting strategy to anchor activatable photosensitizers on the plasma membrane provides a simple but effective method for enhancing the therapeutic efficacy of photosensitizers in anticancer therapy.

Polyprodrug with glutathione depletion and cascade drug activation for multi-drug resistance reversal.

High intracellular glutathione (GSH) levels play an important role in multidrug resistance (MDR) in cancer cells. It remains challenging to develop a drug delivery system that is simultaneously capable of GSH depletion and drug activation for multidrug resistance reversal. Herein, we designed a polyprodrug (denoted as PSSD) based on poly(disulfide) conjugated with doxorubicin (DOX) on the polymer side chains that exhibits GSH depletion and cascade DOX activation for drug resistance reversal. The poly(disulfide) backbone with a high disulfide density depletes intracellular antioxidant GSH via the disulfide-thiol exchange reaction to disrupt intracellular redox homeostasis in cells. Simultaneously, DOX can be activated through a cascade reaction, and degradation of the poly(disulfide) backbone further facilitates its drug release. Therefore, poly(disulfide) can be used as a GSH scavenger to reverse MDR as well as a prodrug backbone to target high intracellular GSH levels in cancer cells, providing a general strategy for drug resistance reversal.

The effect of HES130/0.4 sodium chloride solution on kidney function following early fluid resuscitation in shock patients.

BACKGROUND: Doctors often use a small dose of hydroxyethyl starch (HES) 130/0.4 sodium chloride solution in the emergency room; however, its effect on kidney function remains controversial. This study aimed to evaluate the effect of a small dose of HES130/0.4 sodium chloride solution on kidney function in shock patients during early fluid resuscitation. METHODS: This cohort study retrospectively analyzed the data of 129 shock patients requiring fluid resuscitation who had been admitted to the Emergency Department of the Affiliated Hospital of Nantong University from January 2019 to December 2020. Patients were divided into the observation group (n=40) and control group (n=89) according to the type of fluid resuscitation. In relation to the fluid resuscitation treatment, the observation group was treated with crystalloid solution, while the control group was treated with crystalloid and HES130/0.4 sodium chloride solution. To further explore the effect of a small dose of HES130/0.4 sodium chloride solution, the patients were further divided into the following 4 groups based on the specific fluid administered: (I) the HES(+), lactated Ringer's (LR)(+) group (n=85); (II) the HES(+), LR(-) group (n=4); (III) the HES(-), LR(+) group (n=31); and (IV) the HES(-), LR(-) group (n=9). The outcomes were in-hospital mortality and changes in creatinine (CR) level after fluid resuscitation. RESULTS: There were no significant differences in the in-hospital mortality rates between the observation and control groups (P=0.343). The CR levels of patients in the control and HES(+), LR(+) groups were reduced after fluid resuscitation (P=0.034; P=0.028). There was no significant change in patients' CR levels in the HES(+), LR(-) group after fluid resuscitation (P=0.999). CONCLUSIONS: Administering a small dose of HES 130/0.4 sodium chloride in patients with shock does not appear to affect kidney function and in-hospital mortality; however, these findings should be considered exploratory, and further studies should be conducted to confirm these results.

A clinical study of preoperative carbohydrate administration to improve insulin resistance in patients with multiple injuries.

BACKGROUND: The purpose of this study was to investigate the tolerance and safety of carbohydrate administration to patients with multiple injuries prior to surgery, and to analyze the effects of carbohydrate intake on their immediate insulin resistance (IR), postoperative complications, and length of hospital stay. METHODS: A total of 125 patients with mild multiple injuries who were admitted to the Emergency Surgery Department of Affiliated Hospital of Nantong University for elective surgery were randomized to administration of either placebo or carbohydrate. Finally, 82 patients (male: 39, female: 43) successfully completed the experiment and collected data. Preoperative general condition, subjective comfort, blood glucose concentration, serum insulin and insulin resistance index (IR) were studied. RESULTS: The two groups of patients matched in gender, age, body mass index (BMI) (P>0.05). Patients in CHO group treated with carbohydrates three hours before surgery compared with patients treated with preoperative specification. The thirst, hunger and anxiety of the patients in the CHO group were significantly relieved (P<0.05). Blood glucose concentration, serum insulin, and IR were much lower in the CHO group (P<0.05). CONCLUSIONS: It is a relatively safe approach that patients took carbohydrates 3 hours before surgery, and there was no statistically significant difference in the incidence of postoperative aspiration. Taking carbohydrates before surgery can not only relieve preoperative discomfort, but also reduce postoperative insulin resistance, which is helpful to avoid postoperative metabolic disorder and speed up recovery.

The correlations between the serum expression of miR-206 and the severity and prognosis of sepsis.

BACKGROUND: An accurate assessment of the severity and prognosis of sepsis, especially septic shock, is vital for the tailored treatment of this condition. miRNA participates in the inflammatory response and cell apoptosis and regulates inflammation-related signaling pathways. Immune disorders often accompany sepsis. Since serum miRNA expression is superior to traditional biological markers in terms of sensitivity and specificity, its role in the assessment of sepsis has increasingly been recognized. METHODS: Serum miRNAs were extracted from septic patients and healthy individuals by using the ultracentrifugation method. The differential expressions of miRNAs in the serum samples were detected by high-throughput sequencing technology. The differentially expressed miRNAs between the two groups were analyzed by bioinformatics. The quantitative polymerase chain reaction real-time polymerase chain reaction (qRT-PCR) was used to amplify the sample size to verify the results and to screen the highly-expressed miR206 in septic patients. Subsequently, serum samples were collected from 63 septic patients, and 30 patients with septic shock and qRT-PCR were performed to analyze the expression of miR-206. These 93 patients were divided into the miR-206 low-expression group and miR-206 high-expression group according to miR206 expression level. The potential correlations between the miR-206 expression and the clinical data were analyzed by using SPSS 25.0. RESULTS: Serum miRNA expression significantly differed between septic patients and healthy individuals. High-throughput sequencing results showed that, compared with those in healthy individuals, 29 miRNA molecules were down-regulated, and 25 molecules were up-regulated in the serum samples of septic patients. qRT-PCR identified the significantly up-regulated miR-206 in septic patients. qRT-PCR also showed significantly higher miR-206 expression levels in patients with septic shock than in septic patients. Furthermore, we observed a significantly longer prothrombin time and activated partial thromboplastin time, and significantly higher SOFA score, APACHE-II score, and in-hospital mortality rate. miR-206 was positively correlated with SOFA sore and APACHE-II score. CONCLUSIONS: Serum miR-206 expression is positively correlated with the severity and prognosis of sepsis. Thus, it may be a potential biomarker for assessing the severity and prognosis of sepsis, although the specific mechanism warrants further investigations.

Preparation of doxorubicin-loaded collagen-PAPBA nanoparticles and their anticancer efficacy in ovarian cancer.

BACKGROUND: The aims of this study were to prepare the collagen-poly (3-acrylamidophenylboronic acid) nanoparticles (collagen-PAPBA NPs) encapsulating doxorubicin (DOX) and research their anticancer efficacy in ovarian cancer. METHODS: Collagen-PAPBA NPs were prepared, and their morphology and stability morphology were observed by transmission electron microscopy (TEM) and dynamic light scattering system (DLS). Preparation of doxorubicin-loaded Collagen-PAPBA NPs (DOX-loaded NPs) were then prepared, and the drug-loading content, encapsulation efficiency, and in vitro drug-release profiles were calculated. The morphology of DOX-loaded NPs was also observed by DLS, in vitro cytotoxicity to A2780 cells was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, in vitro antitumor activity on A2780 cells was observed by immunofluorescence, and in vivo antitumor activity was assessed using an experimental BALB/c mice tumor model. RESULTS: DOX-encapsulating collagen-PAPBA NPs were successfully prepared with mediation by biomolecule. The average hydrodynamic diameter of collagen-PAPBA NPs as measured by DLS was about 79 nm, with a homogeneous distribution of size. TEM revealed that nanoparticles were well-dispersed, spherical, and a roughly uniform 75 nm in size. Collagen-PAPBA NPs were quite stable in a wide range of pH and temperature conditions and associated with the concentration of glucose. DLS revealed that the average hydrodynamic diameter of DOX-loaded NPs was about 81.3 nm, with homogeneous distribution of size. TEM revealed that drug-loaded nanoparticles were spherical, well-dispersed, and gad a roughly uniform size of 79 nm. The proportion of DOX loaded into the nanoparticles was 10%, while the encapsulating efficiency was 97%. The result of the releasing test showed that the drug-loaded nanoparticles, as carriers for DOX, had a good sustained-release effect. The cell toxicity experiment showed that the blank NPs had no cytotoxicity to A2780 cells, and that the drug-loaded NPS had good a sustained-release function. They may thus have potential toxic-reducing side effects. CONCLUSIONS: Under the same doses, the drug-loaded NP had a superior inhibitory effect to free DOX on the growth of human ovarian cancer.

Intercellular delivery of bioorthogonal chemical receptors for enhanced tumor targeting and penetration.

Targeted drug delivery using biological ligands can improve the precision of cancer therapy. However, this active targeting strategy is limited in tumor targeting and penetration abilities due to the paucity and heterogeneous distribution of targeted receptors in tumor cells, thus compromising the treatment outcomes. In this study, we developed an alternative active targeting strategy for enhanced tumor targeting and penetration through synthetic nanoparticle-mediated metabolic tumor ligand labeling for intercellular delivery of bioorthogonal chemical receptors combined with in vivo bioorthogonal click chemistry. Briefly, artificial azide-containing ligands were first labeled on perivascular tumor cells by nanoscale metabolic precursors (Az-NPs) via the enhanced permeability and retention (EPR) effect and metabolic engineering of the tumor cells. Through transport by extracellular vesicles (EVs) secreted by perivascular tumor cells, the azide-containing ligands can be autonomously transported intercellularly to adjacent cells and further spread throughout tumor tissues and label bioorthogonal ligands on cells that are not in proximity to blood vessels. Then, water-soluble dibenzocyclooctyne-modified chlorin e6 (DBCO-Ce6) was intravenously injected to react selectively, efficiently and irreversibly with the azide groups on the cell surface through an in vivo bioorthogonal click reaction. Enhanced tumor accumulation and penetration of DBCO-Ce6 was achieved through this strategy, resulting in improved therapeutic efficiency with laser irradiation for photodynamic therapy. Therefore, the artificial azide-containing ligand targeting strategy by nanoparticle-mediated metabolic labeling through the EPR effect combined with bioorthogonal click chemistry may provide an alternative strategy for enhanced tumor targeting and penetration with broad applications.

A General Strategy for Macrotheranostic Prodrug Activation: Synergy between the Acidic Tumor Microenvironment and Bioorthogonal Chemistry.

Prodrugs activated by endogenous stimuli face the problem of tumor heterogeneity. Bioorthogonal prodrug activation that utilizes an exogenous click reaction has the potential to solve this problem, but most of the strategies currently used rely on the presence of endogenous receptors or overexpressed enzymes. We herein integrate the acidic, extracellular microenvironment of a tumor and a click reaction as a general strategy for prodrug activation. This was achieved by using a tumor pH-responsive polymer containing tetrazine groups, which formed unreactive micelles in the blood but disassembled in response to tumor pH. The vinyl ether group on the macrotheranostic prodrug (CyPVE) is activated by the tetrazine groups, which was confirmed by tumor-specific fluorescence activation and phototoxicity restoration. Therefore, the bioorthogonal reactions in the context of the ubiquitous acidic tumor microenvironment can provide a general strategy for bioorthogonal prodrug activation.

Nucleotide Excision Repair Protein Rad23 Regulates Cell Virulence Independent of Rad4 in Candida albicans.

In the pathogenic yeast Candida albicans, the DNA damage response contributes to pathogenicity by regulating cell morphology transitions and maintaining survival in response to DNA damage induced by reactive oxygen species (ROS) in host cells. However, the function of nucleotide excision repair (NER) in C. albicans has not been extensively investigated. To better understand the DNA damage response and its role in virulence, we studied the function of the Rad23 nucleotide excision repair protein in detail. The RAD23 deletion strain and overexpression strain both exhibit UV sensitivity, confirming the critical role of RAD23 in the nucleotide excision repair pathway. Genetic interaction assays revealed that the role of RAD23 in the UV response relies on RAD4 but is independent of RAD53, MMS22, and RAD18RAD4 and RAD23 have similar roles in regulating cell morphogenesis and biofilm formation; however, only RAD23, but not RAD4, plays a negative role in virulence regulation in a mouse model. We found that the RAD23 deletion strain showed decreased survival in a Candida-macrophage interaction assay. Transcriptome sequencing (RNA-seq) and quantitative real-time PCR (qRT-PCR) data further revealed that RAD23, but not RAD4, regulates the transcription of a virulence factor, SUN41, suggesting a unique role of RAD23 in virulence regulation. Taking these observations together, our work reveals that the RAD23-related nucleotide excision pathway plays a critical role in the UV response but may not play a direct role in virulence. The virulence-related role of RAD23 may rely on the regulation of several virulence factors, which may give us further understanding about the linkage between DNA damage repair and virulence regulation in C. albicansIMPORTANCECandida albicans remains a significant threat to the lives of immunocompromised people. An understanding of the virulence and infection ability of C. albicans cells in the mammalian host may help with clinical treatment and drug discovery. The DNA damage response pathway is closely related to morphology regulation and virulence, as well as the ability to survive in host cells. In this study, we checked the role of the nucleotide excision repair (NER) pathway, the key repair system that functions to remove a large variety of DNA lesions such as those caused by UV light, but whose function has not been well studied in C. albicans We found that Rad23, but not Rad4, plays a role in virulence that appears independent of the function of the NER pathway. Our research revealed that the NER pathway represented by Rad4/Rad23 may not play a direct role in virulence but that Rad23 may play a unique role in regulating the transcription of virulence genes that may contribute to the virulence of C. albicans.

microRNA-132 inhibits the proliferation, migration, and invasion of ovarian cancer cells by regulating CT10 oncogenic gene homolog II-related signaling pathways.

BACKGROUND: Despite a large amount of evidence showing the involvement of microRNA-132 (miR-132) in the occurrence and prognosis of many different types of cancer, the role of miR-132 in ovarian cancer and its potential molecular mechanism have yet to be fully explained. METHOD: We studied the biological function and molecular mechanism of miR-132 in ovarian cancer cell lines and clinical tissue samples using quantitative reverse transcription polymerase chain reaction (qRT-PCR), Western blot, Luciferase reporter assay, CCK8 test, colony formation test, and scratch and Transwell assays. RESULTS: The expression level of miR-132 was significantly reduced in ovarian cancer cell lines and clinical tissue samples. When the level of miR-132 was increased, the proliferation, colony-forming, migration, and invasion abilities of ovarian cancer cells were significantly inhibited. We found that miR-132 inhibits the expression of transcription factor CT10 Oncogenic Gene Homologue II (CRKII) through specific targeting of mRNA 3'-UTR. We also observed a significant increase in CRKII expression in ovarian cancer. Notably, CRKII expression was negatively correlated with miR-132 expression in clinical ovarian cancer tissue. Down-regulation of CRKII had a similar inhibitory effect on miR-132 overexpression in ovarian cancer cells, while excessive expression of CRKII reversed the inhibitory effect mediated by the excessive expression of miR-132. CONCLUSIONS: miR-132 inhibits the proliferation, invasion, and migration abilities of ovarian cancer cells through targeting CRKII.

AIEgen based drug delivery systems for cancer therapy.

Fluorescent drug delivery systems provide a means to track drug release, trace processes of translocation, monitor excretion of anticancer agents and predict therapeutic responses. Traditional fluorescent dye-labeled drug delivery systems often suffer from notorious aggregation-caused quenching (ACQ) with greatly impeded imaging performance. The emerging fluorogens with aggregation-induced emission characteristics (AIEgens) have provided an elegant alternative to tackle this challenge. Recently, fluorescent drug delivery systems based on AIEgens which combine fluorescence imaging and drug delivery have been extensively studied for the development of theranostic nanomedicine. In this review, we summarize the recent development of fluorescent drug delivery systems using AIEgens as the signal reporter. This review is organized according to the drug delivery systems, which include: (1) AIEgens as the drug delivery carrier; (2) AIE polymer as the drug delivery carrier; (3) Organic-inorganic AIE nanocarrier for drug delivery; (4) Supramolecular AIE system for drug delivery. Through illustration of their design principles and application examples, we hope to stimulate more interest in the design of more advanced fluorescent drug delivery systems based on AIEgens for cancer therapy.