Intracellularly Synthesized Artificial Exosome Treats Acute Lung Injury.

Acute lung injury (ALI) and its severe form, acute respiratory distress syndrome (ARDS), induce high morbidity and mortality rates, which challenge the present approaches for the treatment of ALI/ARDS. The clinically used photosensitizer verteporfin (VER) exhibits great potential in the treatment of acute lung injury and acute respiratory distress syndrome (ALI/ARDS) by regulating macrophage polarization and reducing inflammation. Nevertheless, its hydrophobic characteristics, nonspecificity, and constrained bioavailability hinder its therapeutic efficacy. In this work, we developed a type of VER-cored artificial exosome (EVM), which was produced by using mesoporous silica nanoparticles (MSNs) to load VER, followed by the exocytosis of internalized VER-MSNs from mouse bone marrow-derived mesenchymal stem cells (mBMSCs) without further modification. Both in vitro and in vivo assessments confirmed the powerful anti-inflammation induced by EVM. EVM also showed significant higher accumulation to inflammatory lungs compared with healthy ones, which was beneficial to the treatment of ALI/ARDS. EVM improved pulmonary function, attenuated lung injury, and reduced mortality in ALI mice with high levels of biocompatibility, exhibiting a 5-fold higher survival rate than the control. This type of artificial exosome emitted near-infrared light in the presence of laser activation, which endowed EVM with trackable ability both in vitro and in vivo. Our work developed a type of clinically used photosensitizer-loaded artificial exosome with membrane integrity and traceability. To the best of our knowledge, this kind of intracellularly synthesized artificial exosome was developed and showed great potential in ALI/ARDS therapy.

MCM8 promotes gastric cancer progression through RPS15A and predicts poor prognosis.

BACKGROUND: Gastric cancer (GC) is the fourth leading cause of cancer-related death worldwide. Minichromsome maintenance proteins family member 8 (MCM8) assists DNA repair and DNA replication. MCM8 exerts tumor promotor function in multiple digestive system tumors. MCM8 is also considered as a potential cancer therapeutic target. METHODS: Bioinformatics methods were used to analyze MCM8 expression and clinicopathological significance. MCM8 expression was detected by immunohistochemistry (IHC) staining and qRT-PCR. MCM8 functions in GC cell were explored by Celigo cell counting, colony formation, wound-healing, transwell, and annexin V-APC staining assays. The target of MCM8 was determined by human gene expression profile microarray. Human phospho-kinase array kit evaluated changes in key proteins after ribosomal protein S15A (RPS15A) knockdown. MCM8 functions were reassessed in xenograft mouse model. IHC detected related proteins expression in mouse tumor sections. RESULTS: MCM8 was significantly upregulated and predicted poor prognosis in GC. High expression of MCM8 was positively correlated with lymph node positive (p < 0.001), grade (p < 0.05), AJCC Stage (p < 0.001), pathologic T (p < 0.01), and pathologic N (p < 0.001). MCM8 knockdown inhibited proliferation, migration, and invasion while promoting apoptosis. RPS15A expression decreased significantly after MCM8 knockdown. It was also the only candidate target, which ranked among the top 10 downregulated differentially expressed genes (DEGs) in sh-MCM8 group. RPS15A was identified as the target of MCM8 in GC. MCM8/RPS15A promoted phosphorylation of P38α, LYN, and p70S6K. Moreover, MCM8 knockdown inhibited tumor growth, RPS15A expression, and phosphorylation of P38α, LYN, and p70S6K in vivo. CONCLUSIONS: MCM8 is an oncogene and predicts poor prognosis in GC. MCM8/RPS15A facilitates GC progression.

Regioselective Homolytic C2-H Borylation of Unprotected Adenosine and Adenine Derivatives via Minisci Reaction.

A Minisci-type borylation of unprotected adenosine, adenine nucleotide, and adenosine analogues was successfully achieved through photocatalysis or thermal activation. Despite the challenges posed by the presence of two potential reactive sites (C2 and C8) in the purine motif, the unique nucleophilic amine-ligated boryl radicals effortlessly achieved excellent C2 site selectivity and simultaneously avoided the formation of multifunctionalized products. This protocol proved effective for the late-stage borylation of some important biomolecules as well as a few antiviral and antitumor drug molecules, such as AMP, cAMP, Vidarabine, Cordycepin, Tenofovir, Adefovir, GS-441524, etc. Theoretical calculations shed light on the site selectivity, revealing that the free energy barriers for the C2-Minisci addition are further lowered through the chelation of additive Mg2+ to N3 and furyl oxygen. This phenomenon has been confirmed by an IGMH analysis. Preliminary antitumor evaluation, derivation of the C2-borylated adenosine to other analogues with high-value functionalities, along with the CuAAC click reactions, suggest the potential application of this methodology in drug molecular optimization studies and chemical biology.

Quercetin induces ferroptosis in gastric cancer cells by targeting SLC1A5 and regulating the p-Camk2/p-DRP1 and NRF2/GPX4 Axes.

Quercetin (Quer) is a natural flavonoid known for its inhibitory effects against various cancers. However, the mechanism by which Quer inhibits gastric cancer (GC) has not yet been fully elucidated. Ferroptosis, a mode of programmed cell death resulting from lipid peroxidation, is regulated by abnormalities in the antioxidant system and iron metabolism. Through flow cytometry and other detection methods, we found that Quer elevated lipid peroxidation levels in GC cells. Transmission electron microscopy confirmed an increase in ferroptosis in Quer-induced GC. We demonstrated that Quer inhibits SLC1A5 expression. Molecular docking revealed Quer's binding to SLC1A5 at SER-343, SER-345, ILE-423, and THR-460 residues. Using immunofluorescence and other experiments, we found that Quer altered the intracellular ROS levels, antioxidant system protein expression levels, and iron content. Mechanistically, Quer binds to SLC1A5, inhibiting the nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2), resulting in decreased xCT/GPX4 expression. Quer/SLC1A5 signaling activated p-Camk2, leading to upregulated p-DRP1 and enhanced ROS release. Additionally, Quer increased the intracellular iron content by inhibiting SLC1A5. These three changes collectively led to ferroptosis in GC cells. In conclusion, Quer targets SLC1A5 in GC cells, inhibiting the NRF2/xCT pathway, activating the p-Camk2/p-DRP1 pathway, and accelerating iron deposition. Ultimately, Quer promotes ferroptosis in GC cells, inhibiting GC progression. Overall, our study reveals that Quer can potentially impede GC progression by targeting SLC1A5, offering novel therapeutic avenues through the modulation of ferroptosis and iron homeostasis.

Enhanced ultrasound-guided versus non-enhanced ultrasound-guided percutaneous needle biopsy in tissue cellularity of lung malignancies: a propensity score matched study.

Background: Though ultrasound-guided percutaneous lung needle biopsy (US-PLNB) is a first-line small biopsy method for peripheral lung lesions, quality of cellularity in specimens obtained via US-PLNB is uncertain. This study investigated the accuracy, sensitivity, and cellularity of US-PLNB. It examined the ability of contrast-enhanced ultrasound (CEUS) to improve the effectiveness of US-PLNB. Methods: We retrospectively analyzed all data of patients with subpleural lung lesions who underwent US-PLNB. The cellularity of US-PLNB from malignant lesions included the tumor cell number and proportion. The definition of high-quality cellularity (HQC) was concurrently achieving a tumor cell number ≥400 and a proportion ≥20%. The sensitivity, the actual numbers of tumor cell number/proportion, and the rate of HQC were calculated and compared between the CEUS and non-enhanced US groups after propensity score matching (PSM) with subgroup analyses by lesion size (small lesion ≤30 mm and large lesion >30 mm). Results: A total of 345 patients undergoing 345 US-PLNBs were evaluated, with 3.7±1.1 of punctures on average. There were 201 malignant and 144 benign lesions with a mean size of 43.8±24.1 mm. Among the 201 malignant lesions, 124 cases underwent CEUS and 77 underwent non-enhanced US. The quantity of tumor cells, the proportion of tumor cells, and the rate of HQC in 201 cases of US-PLNB from malignant lesions were 2,862.1±2,288.0, 44.6%±24.5%, and 82.1% [95% confidence interval (CI): 76.6% to 87.1%], respectively. The quantity of tumor cells, the proportion of tumor cells, and rate of HQC were significantly higher in the CEUS group than that in the non-enhanced US group, both in the analysis of overall malignant lesions and in large malignant lesions (all P<0.05). Conclusions: The US-PLNB has high sensitivity and thereby obtains HQC samples for subpleural lung malignant lesions. The CEUS helps improve the rate of HQC and tissue cellularity of lung malignancies.

Diagnostic ability and its influenced factors of ultrasound-guided percutaneous pleural needle biopsy diagnosis for malignant pleural mesothelioma.

Background: Malignant pleural mesothelioma (MPM) is a highly invasive malignant tumor. Ultrasound guidance has the advantages of real-time, convenience and nonradiative. We sought to identify diagnostic value and its influenced factors of ultrasound-guided percutaneous pleural needle biopsy (US-PPNB) for MPM. Methods: Patients who underwent US-PPNB between March 2014 and March 2020 and were finally diagnosed with MPM were retrospectively analyzed. We retrospectively analyzed the US-PPNBs pathological results of all patients clinically confirmed as MPM, and divided US-PPNBs into correctly and incorrectly diagnosed groups. Patient, thoracic, and biopsy variables that affected diagnostic accuracy were assessed. All variables significant on univariate analyses were subjected to multivariate logistic regression to identify significant predictors of diagnostic accuracy. We derived cutoffs for all significant continuous variables and used the Mantel-Haenszel test to determine whether the diagnostic accuracy of US-PPNB for MPM increased with pleural thickness. Results: In total, 49 patients with clinically confirmed MPM underwent US-PPNB; 37 diagnoses were correct and 12 were incorrect (accuracy = 75.5%). The pleura was significantly thicker in the correctly diagnosed group (p < 0.001). The pleural thickness cutoff was 4.15 mm and diagnostic accuracy increased with pleural thickness grade (p for trend <0.05). The diagnostic accuracy was significantly higher when 16-G rather than 18-G biopsy needles were used (p < 0.05). Multivariate logistic regression showed that pleural thickness (odds ratio: 17.2, 95% confidence interval: 2.8-104.1, p = 0.002) and needle size (odds ratio: 6.8, 95% confidence interval: 1.0-44.5, p = 0.044) independently predicted diagnostic accuracy. Conclusion: US-PPNB afforded high MPM diagnostic accuracy, and pleural thickness and needle size significantly impacted accuracy.

A Comparison of the Performances of Artificial Intelligence System and Radiologists in the Ultrasound Diagnosis of Thyroid Nodules.

AIMS: The purpose of this paper is to prospectively evaluate the performance of an artificial intelligence (AI) system in diagnosing thyroid nodules and to assess its potential value in comparison with the performance of radiologists with different levels of experience, as well as the factors affecting its diagnostic accuracy. BACKGROUND: In recent years, medical imaging diagnosis using AI has become a popular topic in clinical application research. OBJECTIVE: This study aimed to evaluate the performance of an AI system in diagnosing thyroid nodules and compare it with the performance levels of different radiologists. METHODS: This study involved 426 patients screened for thyroid nodules at the First Affiliated Hospital of Guangzhou Medical University between July 2017 and March 2019. All of the nodules were evaluated by radiologists with various levels of experience and an AI system. The diagnostic performances of two junior and two senior radiologists, an AI system, and an AI-assisted junior radiologist were compared, as were their diagnostic results with respect to nodules of different sizes. RESULTS: The senior radiologists, the AI system, and the AI-assisted junior radiologist performed better than the junior radiologist (p < 0.05). The area under the curves of the AI system and the AI-assisted junior radiologist were similar to the curve of the senior radiologists (p > 0.05). The diagnostic results concerning the two nodule sizes showed that the diagnostic error rates of the AI system, junior radiologists, and senior radiologists for nodules with a maximum diameter of ≤1 cm (Dmax ≤ 1 cm) were higher than those for nodules with a maximum diameter of 1 cm (Dmax > 1 cm) (23.4% vs. 12.1%, p = 0.002; 26.6% vs. 7.3%, p < 0.001; and 38.3% vs. 14.6%, p < 0.001). CONCLUSION: The AI system is a decision-making tool that could potentially improve the diagnostic efficiency of junior radiologists. Micronodules with Dmax ≤ 1cm were significantly correlated with diagnostic accuracy; accordingly, more micronodules of this size, in particular, should be added to the AI system as training samples. Other: The system could be a potential decision-making tool for effectively improving the diagnostic efficiency of junior radiologists in the community.

The Binding Technique for Endoscopic Spontaneous Intracerebral Hemorrhage Evacuation.

BACKGROUND: Recently many different endoscopic techniques have been used in spontaneous intracerebral hemorrhage evacuation. However, most of these techniques require expensive special equipment or a well-coordinated assistant. We present a simple and effective binding technique for endoscopic hemorrhage evacuation, which is especially useful during emergency treatment and suitable for use in less-developed areas. METHODS: Our goal was to achieve easy and accurate hemostasis when using an endoscopic technique in which a single surgeon could operate 3 instruments (endoscope, suction tube, and bipolar forceps) with both hands simultaneously in the transparent tubular retractor. This modification, which we called the binding technique, was achieved by bundling the endoscope and suction tube with a sterile rubber band. RESULTS: We performed the binding technique for endoscopic removal of hematoma in 6 patients, including 3 basal ganglia hemorrhages, 2 brain lobe hemorrhages, and 1 cerebellar hemorrhage. The mean operative time was 117.5 minutes (range, 96-155 minutes). One patient died of postoperative delayed brainstem infarction. The Glasgow Outcome Scale score at 3 months was 5 in 3 patients, 3 in 2 patients and 1 (death) in 1 patient. The modified Rankin Scale score at 3 months was 0 in 3 patients, 4 in 2 patients, and 6 (death) in 1 patient. CONCLUSIONS: The binding technique is a modification that allows a single surgeon to achieve easy and accurate hemostasis in endoscopic surgery of intracerebral hematomas. This technique is easy to learn and suitable for emergency surgery, especially in less developed areas.

Direct Detection of Pulmonary Fibrosis by Near-Infrared-Responsive Biomimetic Platelets.

BACKGROUND: Pulmonary fibrosis (PF) is a fatal lung disease and affects over 5 million patients worldwide. Precise and early detection of PF is of pivotal importance to slow the disease progression. However, there are currently no effective tools to detect PF directly. PURPOSE: This study aimed to develop an imaging modality to detect PF directly. Excessive collagen deposition is the hallmark of PF. Herein, we developed a novel PF diagnostic agent, namely PVD (platelets-derived nanovesicles labeled with dye), by utilizing near-infrared (NIR)-responsive biomimetic platelets that specifically recognize collagen. METHODS: In brief, platelets membrane was extracted from purified platelets by freeze/thaw and formed to PVD nanovesicles via sonication and extrusion, when loaded with DiR dye. Red blood cells membrane loaded with DiR was prepared in the same way as PVD to form RVD as control. Collagen self-assembled on microplates was used as an in vitro collagen fibrils model and monocrotaline-induced rats were used as an in vivo PF model. RESULTS: We demonstrated that PVD, but not RVD nor other controls, could bind collagen both in vitro and in vivo, and directly detect pulmonary fibrosis in vivo and ex vivo at the early PF stage. CONCLUSION: Collectively, PVD is a versatile NIR-responsive probe for the direct visualization of collagen, and can be particularly helpful in direct detecting PF. To the best of our knowledge, PVD is the first report of a NIR probe for the direct detection of pulmonary fibrosis.

A Novel Biomimetic Nanoprobe as a Photoacoustic Contrast Agent.

Specific detection of tumors is of pivotal importance to cancer prevention and therapy yet a big challenge. Photoacoustic imaging (PAI) as an emerging non-invasive modality has shown great potential in biomedical and clinical applications. The performance of PAI largely depends on the light-absorption coefficient of the imaged tissue and the PAI contrast agent being used, either endogenously or exogenously. The exogenous contrast agents developed so far have greatly helped to improve PAI, but still have some limitations, such as lack of targeting capacity and easy clearance by the host immune system. Herein, we fabricated a biomimetic nanoprobe with cell membrane coating as a novel PAI contrast agent, namely, MPD [membrane-coated poly(lactic-co-glycolic acid) (PLGA)/dye]. In brief, the organic dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbocyanine iodide (DiR) was encapsulated by the Food and Drug Administration-approved polymer, poly(lactic-co-glycolic acid) (PLGA), to form polymer nanoparticles by emulsification. The nanoparticles are further coated with the cancer cell membrane to form MPD. MPD has outstanding biocompatibility, tumor specificity, and in vivo stability. Thus, MPD is a versatile NIR-I theranostic nanoplatform for PAI-guided cancer diagnosis and therapy.

Hemoptysis complicating ultrasound-guided transthoracic needle lung biopsy: air bronchial sign is a risk predictor.

BACKGROUND: Hemoptysis is the most frequently reported complication of ultrasound-guided transthoracic needle lung biopsy (US-TTLB). However, factors influencing the occurrence of hemoptysis as a result of US-TTLB remain uncertain. Therefore, the aim of this study was to evaluate the incidence of hemoptysis as a complication of US-TTLB and to identify the related risk factors. METHODS: We retrospectively analyzed all data of patients who underwent US-TTLB from February 2013 through December 2016. The incidence, severity, and treatment of hemoptysis in each case were carefully recorded. Study variables were classified into patient-related factors (age, sex, smoking history, pulse oxygen saturation, laboratory tests and emphysema), biopsy-related factors (use of contrast agent, number of punctures and operators), and lesion-related factors (lesion location, size, pathology, length of puncture path and the grade of air bronchial sign). Univariate and multivariate logistic regression analyses were performed to analyze the risk factors of hemoptysis. We investigated whether incidence of hemoptysis increased according to increased grade of air bronchial sign by Mantel-Haenszel test. RESULTS: A total of 209 patients were evaluated. Hemoptysis occurred in 20 of the 209 patients (9.6%). In univariate analysis, the lesion pathology (P=0.037) and grade of air bronchial sign (P<0.001) were statistically significant factors between the hemoptysis group and the non-hemoptysis group. In multivariate analysis, the presence of multi-air bronchogram in sonographic image (odds ratio =8.946; 95% confidence interval: 2.873-27.863; P<0.001) was a statistically significant predictive risk factor for hemoptysis complicating US-TTLB. There was a significant tendency for incidence of hemoptysis with the grade of air bronchial sign (P<0.001). CONCLUSIONS: We found that the rate of hemoptysis complicating US-TTLB was 9.6% and the severity of hemoptysis was not serious. Target lesion without air bronchogram is a safety sign, minor bronchogram means relatively low-risk, while multiple bronchogram is a highly dangerous ultrasound sign of hemoptysis.

Ultrasound-guided pleural cutting needle biopsy: accuracy and factors influencing diagnostic yield.

BACKGROUND: The aim of this study was to retrospectively investigate the diagnostic accuracy of ultrasound-guided pleural cutting needle biopsy (US-guided PCNB) and the potential factors influencing diagnostic yield. METHODS: From July 2014 to June 2016, a total of 147 percutaneous US-guided PCNBs in 144 patients were retrospectively reviewed. The final diagnosis was confirmed by histopathological analysis and follow-up. We calculated diagnostic accuracy, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) and divided all cases into group of correct diagnoses (true-positive and true-negative cases) and group of incorrect diagnoses (false-positive, false-negative, and inconclusive cases). Univariate and multivariate logistic regression analyses were performed to analyze the differences of influencing factors (patient, pleura, and biopsy-associated factors) in the between the two groups. RESULTS: Seven patients were excluded because of loss to follow-up. A total of 140 cases were ultimately included (105 males and 35 females). There were 105 cases in the correct diagnosis group, and 35 cases in the incorrect diagnosis group. The overall accuracy of US-PCNB was 75.0% and the sensitivity, specificity, PPV, NPV in malignant diagnosis were 58.1%, 99.0%, 96.2%, and 84.2%, respectively. On univariate analysis, variables affecting diagnostic accuracy of US-PCNB were the pleural thickness (<3 mm in thickness 61.0%, ≥3 mm in thickness 85.2%; P=0.001), morphology (non-nodular pleura 71.4%, nodular pleura 95.2%; P=0.026), and needle size (18 G 69.1%, 16 G 87.0%; P=0.022). Finally multivariate logistic regression demonstrated that pleural thickness [odds ratio (OR): 0.278, P=0.003] and needle size (OR: 0.291, P=0.018) independently predicted diagnostic accuracy. CONCLUSIONS: Pleural thickness and the size of the biopsy needle were significantly correlated with the diagnostic yield.

Utility of contrast-enhanced ultrasound with SonoVue in biopsy of small subpleural nodules.

OBJECTIVES: This study aimed to evaluate the diagnostic accuracy and complication rates of contrast-enhanced ultrasound (CEUS)-guided biopsy of small subpleural nodules with SonoVue. METHODS: CEUS-guided biopsies with SonoVue and conventional ultrasound were performed to determine nodule size, texture and biopsy route. After baseline ultrasonography, all patients received an intravenous injection of 4 mL of SonoVue, followed by 5 mL of saline flush. CEUS was obtained using a convex probe and contrast-specific imaging software. The lesion was observed using a contrast agent. Biopsies were performed during real-time visualisation of the target lesion. RESULTS: A total of 51 patients (34 males and 17 females; average age, 54.8 ± 5.8 years) with subpleural nodules were enrolled. The median nodule size was 1.92 ± 0.75 cm (0.9-2.5 cm). Forty-eight of 51 procedures (94.1%) provided adequate material for histological analysis. Thirty patients (62.5%) were malignant and 18 patients (37.5%) were benign at the definitive diagnosis. The true positive and true negative result were 28 (58.3%) and 18 (37.5%), no false positive result was seen and two (4.2%) provided a false negative result. The sensitivity, specificity, positive and negative predictive values for the malignant diagnosis were 93.3, 100, 100 and 90%, respectively. The diagnostic accuracy was 95.8% (46/48), the standard error and the 95% CI were 2.8% and 86%-99%. An asymptomatic pneumothorax was present in one patient with no chest tube placement required. A small amount of hemoptysis was observed in another patient, which stopped spontaneously without treatment. CONCLUSIONS: CEUS-guided biopsy with SonoVue exhibits high diagnostic accuracy and low complication rates. It is especially advantageous for biopsies of small subpleural nodules.

Ultrasound microbubble contrast agent-mediated suicide gene transfection in the treatment of hepatic cancer.

The aim of this study was to investigate the role of ultrasound microbubble contrast agent-mediated suicide gene transfection in the treatment of hepatic cancer. We intratumorally injected KDR-TK, AFP-TK and microbubble contrast agent into nude mice prior to ultrasound treatment and administration of prodrugs (GCV and 5-FC). The tumor volume, tumor inhibition rate, survival time and apoptosis of tumor cells was determined. The sizes of subcutaneous hepatic cancers in mice receiving treatment were comparable to those in the control group, and the survival time was similar between the two groups (P>0.05). However, the tumor inhibition rate and the number of apoptotic cells in the treatment group was markedly higher compared with that in the control group (P<0.05). Evident tumor necrosis was absent in both groups, except at the needle tract. Ultrasound therapy following injection of suicide genes and microbubble contrast agents is able to inhibit cancer growth in vivo. This may be attributed to the induction of cancer cell apoptosis.