FLCN-regulated miRNAs suppressed reparative response in cells and pulmonary lesions of Birt-Hogg-Dubé syndrome.

BACKGROUND: Birt-Hogg-Dubé Syndrome (BHDS) characterised by skin fibrofolliculomas, kidney tumour and pulmonary cysts/pneumothorax is caused by folliculin (FLCN) germline mutations. The pathology of both neoplasia and focused tissue loss of BHDS strongly features tissue-specific behaviour of the gene. Isolated cysts/pneumothorax is the most frequent atypical presentation of BHDS and often misdiagnosed as primary spontaneous pneumothorax (PSP). Deferential diagnosis of BHDS with isolated pulmonary presentation (PSP-BHD) from PSP is essential in lifelong surveillance for developing renal cell carcinoma. METHODS: The expression profiles of microRNAs (miRNAs) in cystic lesions of PSP-BHD and PSP were determined via microarray. The selected upregulated miRNAs were further confirmed in the plasma of an expanded cohort of PSP-BHD patients by reverse transcription quantitative PCR (RT-qPCR). Their diagnostic accuracy was evaluated. Moreover, the cellular functions and targeted signalling pathways of FLCN-regulated miRNAs were assessed in various cell lines and in the lesion tissue contexts. RESULTS: Cystic lesions of PSP-BHD and PSP showed different miRNAs profiles with a significant upregulation of miR-424-5p and let-7d-5p in PSP-BHD. The combination of the two effectively predicted BHDS patients. In vitro studies revealed a suppressive effect of FLCN on miR-424-5p and let-7d-5p expressions specifically in lung epithelial cells. The ectopic miRNAs triggered epithelial apoptosis and epithelial transition of mesenchymal cells and suppressed the reparative responses in cells and tissues with FLCN deficiency. CONCLUSION: The upregulation of miR-424-5p and let-7d-5p by FLCN deficiency occurred in epithelial cells and marked the PSP-BHD condition, which contributed to a focused degenerative pathology in the lung of PSP-BHD patients.

A CsTu-TS1 regulatory module promotes fruit tubercule formation in cucumber.

The fruit epidermal features such as the size of tubercules are important fruit quality traits for cucumber production. But the mechanisms underlying tubercule formation remain elusive. Here, tubercule size locus CsTS1 was identified by map-based cloning and was found to encode an oleosin protein. Allelic variation was identified in the promoter region of CsTS1, resulting in low expression of CsTS1 in all 22 different small-warty or nonwarty cucumber lines. High CsTS1 expression levels were closely correlated with increased fruit tubercule size among 44 different cucumber lines. Transgenic complementation and RNAi-mediated gene silencing of CsTS1 in transgenic cucumber plants demonstrated that CsTS1 positively regulates the development of tubercules. CsTS1 is highly expressed in the peel at fruit tubercule forming and enlargement stage. Auxin content and expression of three auxin signalling pathway genes were altered in the 35S:CsTS1 and CsTS1-RNAi fruit tubercules, a result that was supported by comparing the cell size of the control and transgenic fruit tubercules. CsTu, a C2 H2 zinc finger domain transcription factor that regulates tubercule initiation, binds directly to the CsTS1 promoter and promotes its expression. Taken together, our results reveal a novel mechanism in which the CsTu-TS1 complex promotes fruit tubercule formation in cucumber.

A rapid NGS strategy for comprehensive molecular diagnosis of Birt-Hogg-Dubé syndrome in patients with primary spontaneous pneumothorax.

BACKGROUND: Primary spontaneous pneumothorax (PSP) or pulmonary cysts is one of the manifestations of Birt-Hogg-Dube syndrome (BHDS) that is caused by heterozygous mutations in FLCN gene. Most of the mutations are SNVs and small indels, and there are also approximately 10 % large intragenic deletions and duplications of the mutations. These molecular findings are generally obtained by disparate methods including Sanger sequencing and Multiple Ligation-dependent Probe Amplification in the clinical laboratory. In addition, as a genetically heterogeneous disorder, PSP may be caused by mutations in multiple genes include FBN1, COL3A1, CBS, SERPINA1 and TSC1/TSC2 genes. For differential diagnosis, these genes should also be screened which makes the diagnostic procedure more time-consuming and labor-intensive. METHODS: Forty PSP patients were divided into 2 groups. Nineteen patients with different pathogenic mutations of FLCN previously identified by conventional Sanger sequencing and MLPA were included in test group, 21 random PSP patients without any genetic screening were included in blinded sample group. 7 PSP genes including FLCN, FBN1, COL3A1, CBS, SERPINA1 and TSC1/TSC2 were designed and enriched by Haloplex system, sequenced on a Miseq platform and analyzed in the 40 patients to evaluate the performance of the targeted-NGS method. RESULTS: We demonstrated that the full spectrum of genes associated with pneumothorax including FLCN gene mutations can be identified simultaneously in multiplexed sequence data. Noteworthy, by our in-house copy number analysis of the sequence data, we could not only detect intragenic deletions, but also determine approximate deletion junctions simultaneously. CONCLUSIONS: NGS based Haloplex target enrichment technology is proved to be a rapid and cost-effective screening strategy for the comprehensive molecular diagnosis of BHDS in PSP patients, as it can replace Sanger sequencing and MLPA by simultaneously detecting exonic and intronic SNVs, small indels, large intragenic deletions and determining deletion junctions in PSP-related genes.

FLCN intragenic deletions in Chinese familial primary spontaneous pneumothorax.

Primary spontaneous pneumothorax (PSP) is a significant clinical problem, affecting tens of thousands patients annually. Germline mutations in the FLCN gene have been implicated in etiology of familial PSP (FPSP). Most of the currently identified FLCN mutations are small indels or point mutations that detected by Sanger sequencing. The aim of this study was to determine large FLCN deletions in PSP families that having no FLCN sequence-mutations. Multiplex ligation-dependent probe amplification (MLPA) assays and breakpoint analyses were used to detect and characterize the deletions. Three heterozygous FLCN intragenic deletions were identified in nine unrelated Chinese families including the exons 1-3 deletion in two families, the exons 9-14 deletion in five families and the exon 14 deletion in two families. All deletion breakpoints are located in Alu repeats. A 5.5 Mb disease haplotype shared in the five families with exons 9-14 deletion may date the appearance of this deletion back to approximately 16 generations ago. Evidences for founder effects of the other two deletions were also observed. This report documents the first identification of founder mutations in FLCN, as well as expands mutation spectrum of the gene. Our findings strengthen the view that MLPA analysis for intragenic deletions/duplications, as an important genetic testing complementary to DNA sequencing, should be used for clinical molecular diagnosis in FPSP.

Biological removal of antiandrogenic activity in gray wastewater and coking wastewater by membrane reactor process.

A recombinant human androgen receptor yeast assay was applied to investigate the occurrence of antiandrogens as well as the mechanism for their removal during gray wastewater and coking wastewater treatment. The membrane reactor (MBR) system for gray wastewater treatment could remove 88.0% of antiandrogenic activity exerted by weakly polar extracts and 97.3% of that by moderately strong polar extracts, but only 32.5% of that contributed by strong polar extracts. Biodegradation by microorganisms in the MBR contributed to 95.9% of the total removal. After the treatment, the concentration of antiandrogenic activity in the effluent was still 1.05 µg flutamide equivalence (FEQ)/L, 36.2% of which was due to strong polar extracts. In the anaerobic reactor, anoxic reactor, and membrane reactor system for coking wastewater treatment, the antiandrogenic activity of raw coking wastewater was 78.6 mg FEQ/L, and the effluent of the treatment system had only 0.34 mg FEQ/L. The antiandrogenic activity mainly existed in the medium strong polar and strong polar extracts. Biodegradation by microorganisms contributed to at least 89.2% of the total antiandrogenic activity removal in the system. Biodegradation was the main removal mechanism of antiandrogenic activity in both the wastewater treatment systems.

Continuous operation of nano-catalytic ozonation using membrane separation coupling system: Influence factors and mechanism.

The application of nano-catalysts in improving the ozonation removal efficiency for refractory organic compounds has been extensively investigated. However, cost-effective nano-catalysts separation remains a challenge. In this study, membrane separation processes were employed to separate nano-MgO catalysts from an ozonation system. A continuous nano-catalytic ozonation membrane separation (nCOMS) coupling system was successfully constructed for treating quinoline. The results showed that long hydraulic retention time (HRT) and high nano-MgO dosage could improve the quinolone removal efficiency but shorten operation cycles. At the optimal operation conditions of HRT = 4 h and nano-MgO dosage = 0.2 g/L, the nCOMS system achieved a stable quinoline removal efficiency of 85.2% for 240 min running with a transmembrane pressure lower than 10 kPa. The quinoline removal efficiency contribution for ozonation, catalysis and membrane separation was 57.1%, 24.9% and 18.0%, respectively. Compared to ozonation membrane separation system, the fouling rate index of the nCOMS system increased by 60% under optimal conditions, but the irreversible fouling was reduced to 28%. In addition, the nCOMS system exhibited reduced adverse effects of coexisting natural organic matter (NOM) on quinoline removal and membrane fouling. In conclusion, the nCOMS system demonstrated higher quinoline removal efficiency, lower irreversible fouling, and reduced adverse effect of coexisting NOM, thereby signifying its potential for practical applications in advanced treatment of industrial wastewater.

Formation and toxicity contribution of chlorinated and dechlorinated halobenzoquinones from dichlorophenols after ozonation.

Halobenzoquinones (HBQs) are a class of disinfection byproducts with high cytotoxicity and potential carcinogenicity, which have been widely detected in chlorination of drinking water and swimming pool water. However, to date, the formation of HBQs upon ozonation and the HBQ precursors have been overlooked. This study investigated the formation of chlorinated and dechlorinated HBQs from six dichlorophenol (DCP) isomers. The monomeric and dimeric HBQs were identified in all the ozonation effluents, exhibiting 1-100 times higher toxicity levels than their precursors. The sum of detected HBQs intensity had a satisfactory linear relation with the maximum toxic unit (R2 = 0.9657), indicating the primary toxicity contribution to the increased overall toxicity of effluents. Based on density functional theory calculations, when ozone attacks the para carbon to the hydroxyl group of 2,3-DCP, the probability of producing chlorinated HBQs is 80.41 %, indicating that the para carbon attack mainly resulted in the formation of monomeric HBQs. 2,3-dichlorophenoxy radicals were successfully detected in ozonated 2,3-DCP effluent through electron paramagnetic resonance and further validated using theoretical calculation, revealing the formation pathway of dimeric HBQs. The results indicate that chlorinated phenols, regardless of the positions of chlorine substitution, can potentially serve as precursors for both chlorinated and dechlorinated HBQs formation during ozonation.

EGFR co-mutation is associated with the risk of recurrence in invasive lung adenocarcinoma with the micropapillary component.

BACKGROUND: Invasive lung adenocarcinoma (LUAD) patients with the micropapillary (MPP) component tend to have extremely poor prognosis. To optimize clinical outcomes, a better understanding of specific concurrent gene alterations and their impact on the prognosis of patients with the MPP component is necessary. METHOD: A total of 621 Chinese patients with surgically resected invasive LUAD who underwent genetic testing for lung cancer were enrolled in this retrospective study. The genomic profiling of major lung cancer-related genes based on next-generation sequencing (NGS) was carried out on formalin-fixed paraffin-embedded tumor samples. RESULT: Among 621 patients with invasive LUAD, 154 (24.8%, 154/621) had the MPP component. We found that PIK3CA (4.5% vs 1.3%), KRAS (9.1% vs 4.7%), and ROS1 (2.6% vs 0.4%) were more frequent in patients with the MPP component than those without the MPP component (P < 0.05). The co-mutation occurred in 66 patients (10.6%, 66/621), of which 19 patients with the MPP component. Most of them were EGFR co-mutations (89.5%, 17/19), including EGFR and PIK3CA, EGFR and ERBB2, and other types. Patients with the MPP component who harbored EGFR co-mutations showed significantly worse recurrence-free survival (RFS) than single EGFR mutation (median RFS 20.1 vs 30.5 months; hazard ratio [HR]: 8.008; 95% confidence interval [CI]: 1.322-48.508). CONCLUSIONS: Patients with the MPP component harbored the co-mutation of driver genes had a higher risk of recurrence after surgery, especially in patients with EGFR co-mutation. EGFR co-mutation was a significant prognostic factor for RFS in patients with the MPP component.

Biochar assists phosphate solubilizing bacteria to resist combined Pb and Cd stress by promoting acid secretion and extracellular electron transfer.

Microorganisms have difficulty surviving and performing remediation functions in mixed systems with high concentrations of Pb and Cd. Biochar has the potential to assist microorganism remediation as an excellent adsorbent for heavy metals. In this study, pig manure biochar (PMB) was used to assist phosphorus solubilizing bacteria (PSB) to explore the mineralization protection and biofeedback mechanism of biochar on PSB under mixed stress of 1000 mg/L Pb2+ and 500 mg/L Cd2+. The adsorption results showed that the removal of Pb2+ and Cd2+ by PMB+PSB was 148.77% and 72.27% higher than that by PSB. Meanwhile, the non-bioavailable fraction of Cd2+ and acid-soluble fraction of Pb2+ in PMB+PSB were increased by 9% and 3%, respectively. Mineralogical and microbial secretion results confirm that showed that the acidic soluble fraction and non-bioavailable fraction were mostly Pb/Cd-carbonate and Pb/Cd-phosphate. The pore adsorption and precipitation (carbonate) of biochar were able to reduce the exposure of PSB to Pb/Cd and the background stress concentration, thus stimulating the biological positive feedback effect of PSB and forming a microenvironment in the cell periphery. The vesicle detoxification and extracellular polymeric substance protection mechanism of PSB were improved under biochar protection, and the individual size and activity of PSB cells were enhanced. Besides, citric acid release from PSB (28.85% increase) accelerated the dissolution of unstable Cd-carbonate, thereby releasing a large amount of Cd2+ to compete with Pb2+ for PO43-. Thus, the protection of biochar and the positive feedback effect of PSB could reduce the biotoxicity of Cd2+ in the stress system by preferentially forming a stable Cd-phosphate. In addition, the excellent electrical conductivity and organic material adsorption of biochar increased the extracellular electron transport rate of microorganisms, which further accelerated the mineralization and immobilization of Pb2+ and Cd2+, so as to ensure the repair effect of PSB on heavy metals.

TNF-α and IL-8 levels are positively correlated with hypobaric hypoxic pulmonary hypertension and pulmonary vascular remodeling in rats.

The expression status of proinflammatory cytokines in high-altitude pulmonary arterial hypertension (PAH) has been well studied. However, the changes in interleukin (IL)-8 and tumor necrosis factor α (TNF-α) during the reversible changes in pulmonary vascular remodeling (PVR) in PAH after detaching from a hypobaric hypoxic environment have not been elucidated. This investigation elucidated a high-altitude PAH rat model. Then, PAH rats in the high-altitude group were maintained in the high-altitude area, and rats in the low-altitude group returned to the low-altitude area. After 0, 10, 20, and 30 days of PAH modeling, right ventricular systolic pressure (RVSP) and the mean pulmonary arterial pressure (mPAP) were assessed. Right ventricular (RV) hypertrophy was reflected by the ratio of RV/[left ventricle + interventricular septum (S)]. Pathological changes in PVR were accessed by hematoxylin-eosin staining, and medial wall thickness (WT%) and medial wall area (WA%) were measured. TNF-α and IL-8 levels in pulmonary artery tissues and blood were measured with Western blot assay and enzyme-linked immunosorbent assay, respectively. Our results showed that PAH rats exhibited a substantial increase in RVSP and mPAP, RV hypertrophy, PVR, and enhanced generation of TNF-α and IL-8. Then, we found that these pathological changes were gradually aggravated and TNF-α and IL-8 levels were increased in rats in the high-altitude group after 10, 20, and 30 days of PAH modeling. In contrast, the mPAP was decreased and PVR was alleviated in rats in the low-altitude group, accompanying with reduced TNF-α and IL-8 production. In conclusion, our study demonstrated that the generation of TNF-α and IL-8 was also reversible during the reversible changes in PVR after detaching from a hypobaric hypoxic environment. Thus, proinflammatory cytokine TNF-α and IL-8 levels are positively correlated with PVR severity.

Dual-Phasic, Well-Aligned, and Strong Flexible Hydrophobic Ceramic Membranes for Efficient Thermal Insulation in Extreme Conditions.

The inherent brittleness and hydrophilicity of ceramics pose a great challenge to designing a reliable structure that can resist mechanical loads and moisture in extreme conditions with high temperature and high humidity. Here, we report a two-phase hydrophobic silica-zirconia composite ceramic nanofiber membrane (H-ZSNFM) with exceptional mechanical robustness and high-temperature hydrophobic resistance. For the dual-phasic nanofibers, the amorphous silica blocked the connection of zirconia nanocrystals, and the lattice distortion was observed due to Si in the ZrO2 lattice. H-ZSNFM has strong strength (5-8.4 MPa), high hydrophobic temperature resistance (450 °C), high porosity (89%), low density (40 mg/cm3), low thermal conductivity (30 mW/m·K), and excellent thermal radiation reflectivity (90%). By simulating the actual high-temperature and high-humidity environment, 10-mm-thick H-ZSNFMs can reduce the heat source from 1365 to 380 °C and maintain complete hydrophobicity even in a water vapor environment of 350 °C. This means that it has superior insulation and waterproof performance even in a high-temperature water environment. For firefighting clothing, H-ZSNFM displayed waterproof and insulation layers, which have excellent thermal protection performance and achieve incompatibility between water and fire, providing valuable time for fire rescue and a safety line of defense for emergency personnel. This design strategy with mechanical robust and hydrophobic temperature resistance applies to the development of many other types of high-performance thermal insulation materials and presents a competitive material system for thermal protection in extreme conditions.

Redox-directed identification of toxic transformation products during ozonation of aromatics.

The toxicity assessment of transformation products (TPs) formed in oxidative water treatment is crucial but challenging because of their low concentration, structural diversity, and mixture complexity. Here, this study developed a novel redox-directed approach for identification of toxic TPs without the individual toxicity and concentration information. This approach based on sodium borohydride reduction comprised an integrated process of toxicological evaluation, fluorescence excitation-emission matrix characterization, high-resolution mass spectrometry detection, followed by ecological toxicity assessment of identified TPs. The redox-directed identification of primary causative toxicants was experimentally tested for the increased nonspecific toxicity observations in the ozonated effluents of model aromatics. Reduction reaction caused a remarkable decrease in toxicity and increase in fluorescence intensity, obtaining a good linear relation between them. More than ten monomeric or dimeric p-benzoquinone (p-BQ) TPs were identified in the ozonated effluents. The occurrence of the p-BQ TPs was further verified through parallel sodium sulfite reduction and actual wastewater ozonation experiments. In vitro bioassays of luminescent bacteria, as well as in silico genotoxicity and cytotoxicity predictions, indicate that the toxicity of p-BQ TPs is significantly higher than that of their precursors and other TPs. These together demonstrated that the identified p-BQ TPs are primary toxicity contributors. The redox-directed approach facilitated the revelation of primary toxicity contribution, illustrating emerging p-BQs are a concern for aquatic ecosystem safety in the oxidative treatment of aromatics-contaminated wastewater.

Revealing Disparities in Porous Networks Between Yttria Aerogel Assemblies with Nanosheets and Nanoparticles and Their Ultrathermal Insulation and Optical Properties.

Recent advancements have introduced anisotropic structures, particularly 2D nanosheets, into aerogels, resulting in unique morphologies and exceptional properties that differ from those assembled by isotropic nanoparticles. However, exploration of the distinct porous networks and the resulting properties is limited. We focus on rare earth yttria (Y2O3) aerogels as a case in point and demonstrate the synthesis of aerogels with nanosheet and nanoparticle assemblies using elaborative sol-gel chemistry. With the aid of X-ray computed tomography, three-dimensional visualizations of the aerogels provide relative compressive views of the porous network, revealing that the Y2O3 aerogel assembled by nanosheets possesses a hierarchical pore structure characterized by uneven pore distribution, particularly the presence of macropores throughout; in contrast, these consist of nanoparticles exhibiting a relative uniform pore distribution. High-temperature examinations indicate that the nanosheet aerogels are much more stable with a specific surface area of 64 m2·g-1 after being exposed at 1300 °C; meanwhile, the aerogels present durable and efficient thermal insulation performances. The exceptional thermal properties are attributed to the synergistic effects of the nanosheets' crystalline nature and the hierarchical porous network. The nanosheet Y2O3 aerogel also exhibited superior luminescent emission characteristics, further enhancing its potential for various applications. Our findings provide further insights into optimization of the microstructures in nanoporous aerogels, particularly through the utilization of anisotropic nanosheets.

Minigene Assay as an Effective Molecular Diagnostic Strategy in Determining the Pathogenicity of Noncanonical Splice-Site Variants in FLCN.

Primary spontaneous pneumothorax (PSP) or pulmonary cyst is one of the manifestations of Birt-Hogg-Dubé syndrome, which is caused by pathogenic variants in FLCN gene. Genetic testing in patients with PSP identifies a certain number of missense or intronic variants. These variants are usually considered as variants of uncertain significance, whose functional interpretations pose a challenge in clinical genetics. To improve recognition of pathogenic splice-altering variants in FLCN gene, computational tools are used to prioritize potential splice-altering variants and then a hybrid minigene assay is performed to verify the RNA splicing pattern. Herein, variants in FLCN exon 11 and its flanking sequence are focused. Eight variants detected in 11 patients with PSP are evaluated, and six variants are prioritized by in silico tools as potential splice-altering variants of uncertain significance. Four variants (c.1177-5_1177-3delCTC, c.1292_1300+4del, c.1300+4C>T, and c.1300+5G>A) are demonstrated by minigene assay to alter RNA splicing of FLCN, and the last three of them are novel. RT-PCR of patient-derived RNA gives consistent results. Genotype-phenotype correlation analysis in patients with PSP with these variants demonstrates good concordance. Our results underline the importance of RNA analysis, which could provide molecular evidence for pathogenicity of a variant, and provide essential information for the clinical interpretation of variants. Combining the clinical information, a definitive diagnosis could be made.

Extracorporeal membrane oxygenation (ECMO)-assisted surgery for traumatic bronchial rupture: a report of three cases.

Background: Traumatic tracheal rupture is a severe closed chest injury that often causes major respiratory and circulatory disturbances requiring emergency surgery. We have found that veno-venous extracorporeal membrane oxygenation (VV-ECMO) employs lung-protective ventilation strategies to facilitate lung rest, aiming to minimize the risk of ventilator-induced lung injury, while ensuring adequate oxygenation. Case Description: We presented 3 critically ill patients who presented with traumatic bronchial rupture between 2019 and 2021, and underwent emergency thoracic surgery with the help of VV-ECMO. The ECMO support time, the operative time, the duration of postoperative hospital stay, and the postoperative mechanical ventilation time were collected in this study. All patients were successfully treated and discharged home. The duration of surgery ranged from 135 to 180 min, the duration of ECMO use ranged from 98 to 123 h, the duration of postoperative ventilator use ranged from 5 to 8 days, and the duration of postoperative hospital stay ranged from 14 to 30 days. All 3 patients had good postoperative pulmonary re-expansion, with no residual tracheal or bronchial stenosis, and good physical activity following the surgery. Conclusions: We reported successful use of VV-ECMO in critically ill patients with traumatic bronchial rupture presenting in acute respiratory and circulatory failure. Performing emergency surgery with ECMO-assisted support can provide more time to stabilize the patient and ensure the safety of the procedure. However, considering the small sample size of this study, larger cohorts with long-term follow-up data are needed to further evaluate its application.

Deep learning-based solid component measuring enabled interpretable prediction of tumor invasiveness for lung adenocarcinoma.

BACKGROUND: The nature of the solid component of subsolid nodules (SSNs) can indicate tumor pathological invasiveness. However, preoperative solid component assessment still lacks a reference standard. METHODS: In this retrospective study, an AI algorithm was proposed for measuring the solid components ratio in SSNs, which was used to assess the diameter ratio (1D), area ratio (2D), and volume ratio (3D). The radiologist measured each SSN's consolidation to tumor ratio (CTR) twice, four weeks apart. The area under the receiver-operating characteristic (ROC) curve (AUC) was calculated for each method used to discriminate an Invasive Adenocarcinoma (IA) from a non-IA. The AUC and the time cost of each measurement were compared. Furthermore, we examined the consistency of measurements made by the radiologist on two separate occasions. RESULTS: A total of 379 patients (the primary dataset n = 278, the validation dataset n = 101) were included. In the primary dataset, compared to the manual approach (AUC: 0.697), the AI algorithm (AUC: 0.811) had better predictive performance (P =.0027) in measuring solid components ratio in 3D. Algorithm measurement in 3D had an AUC no inferior to 1D (AUC: 0.806) and 2D (AUC: 0.796). In the validation dataset, the AI 3D method also achieved superior diagnostic performance compared to the radiologist (AUC: 0.803 vs 0.682, P =.046). The two measurements of the CTR in the primary dataset, taken 4 weeks apart, have 7.9 % cases in poor consistency. The measurement time cost by the radiologist is about 60 times that of the AI algorithm (P <.001). CONCLUSION: The 3D measurement of solid components using AI, is an effective and objective approach to predict the pathological invasiveness of SSNs. It can be a preoperative interpretable indicator of pathological invasiveness in patients with lung adenocarcinoma.

PFKP confers chemoresistance by upregulating ABCC2 transporter in non-small cell lung cancer.

Background: Chemoresistance is a significant factor contributing to tumor recurrence and treatment failure in non-small cell lung cancer (NSCLC). The phosphofructokinase, platelet (PFKP) is highly expressed in NSCLC and is associated with a poor prognosis. Exploring the molecular mechanism and identifying effective strategies to overcome chemoresistance will have important clinical significance in improving the diagnosis and treatment of NSCLC. Methods: The correlation between PFKP and cisplatin resistance in NSCLC patients was assessed by organoids and immunohistochemistry. The impact of PFKP on the prognosis of NSCLC patients was analyzed using The Cancer Genome Atlas (TCGA) database. In NSCLC cell lines, the expression of PFKP was modulated using lentivirus, and cisplatin sensitivity was assessed by flow cytometry. Subsequently, the therapeutic effect of cisplatin was tested in BALB/c nude mice implanted subcutaneously with tumor cells. We performed luciferase assay and immunohistochemistry (IHC) to investigate the correlation between PFKP and ABCC2 (ATP-binding cassette sub-family C member 2). Results: Overexpression of PFKP was correlated with poorer survival rates in NSCLC patients who received platinum-based chemotherapy. Using NSCLC organoid, we found that the expression of PFKP was elevated in cisplatin (CDDP)-resistant patients with NSCLC. Overexpression of PFKP decreased the sensitivity of NSCLC cells to CDDP, while genetic inhibition of PFKP enhanced CDDP sensitivity both in vitro and in vivo. Furthermore, we found that PFKP upregulated ABCC2 by increasing the levels of phosphorylation of IκBα and nuclear p65 NF-κB subunit protein. Conclusions: PFKP can regulate the expression of ABCC2 through the activation of NF-κB, which in turn promotes chemoresistance in NSCLC. PFKP has the potential to be a personalized therapeutic target for NSCLC patients with chemoresistance.

Invasiveness assessment by artificial intelligence against intraoperative frozen section for pulmonary nodules ≤ 3 cm.

PURPOSE: To investigate the performance of an artificial intelligence (AI) algorithm for assessing the malignancy and invasiveness of pulmonary nodules in a multicenter cohort. METHODS: A previously developed deep learning system based on a 3D convolutional neural network was used to predict tumor malignancy and invasiveness. Dataset of pulmonary nodules no more than 3 cm was integrated with CT images and pathologic information. Receiver operating characteristic curve analysis was used to evaluate the performance of the system. RESULTS: A total of 466 resected pulmonary nodules were included in this study. The areas under the curves (AUCs) of the deep learning system in the prediction of malignancy as compared with pathological reports were 0.80, 0.80, and 0.75 for all, subcentimeter, and solid nodules, respectively. Additionally, the AUC in the AI-assisted prediction of invasive adenocarcinoma (IA) among subsolid lesions (n = 184) was 0.88. Most malignancies that were misdiagnosed by the AI system as benign diseases with a diameter measuring greater than 1 cm (26/250, 10.4%) presented as solid nodules (19/26, 73.1%) on CT. In an exploratory analysis involving nodules underwent intraoperative pathologic examination, the concordance rate in identifying IA between the AI model and frozen section examination was 0.69, with a sensitivity of 0.50 and specificity of 0.97. CONCLUSION: The deep learning system can discriminate malignant diseases for pulmonary nodules measuring no more than 3 cm. The AI model has a high positive predictive value for invasive adenocarcinoma with respect to intraoperative frozen section examination, which might help determine the individualized surgical strategy.

Perioperative Management of Patient with Esophageal Carcinoma and Crigler-Najjar Syndrome Type 2: A Case Report.

Background: Crigler-Najjar syndrome type 2 (CNS-II) is a rare genetic disease that is associated with a lack of uridine diphosphate-glucuronosyltransferase. Esophageal carcinoma is the sixth most common cause of cancer-related death worldwide, for which surgery is the most effective treatment. Reports on patients with both conditions requiring surgery are limited and The impact of hyperbilirubinemia caused by CNS-II on the perioperative period is unknown. Previous studies have found that patients with Crigler-Najjar syndrome have an increased risk of gallstones and related complications, which also poses corresponding challenges to the treatment. Herein, we present a patient with CNS-II who underwent successful thoracoscopic surgery for esophageal carcinoma. Case summary: A 65-year-old male presented to our hospital with a choking sensation after eating. A physical examination showed yellowing of the sclera and skin. The patient manifested persistent jaundice since birth and had visited many hospitals, but the cause remained undiagnosed. We performed genetic testing, which confirmed CNS-II. Gastroscopy indicated esophageal carcinoma. A multidisciplinary team discussion was carried out to determine the appropriate treatment and perioperative management for this patient. The results show that surgical resection was the most appropriate approach. Finally, the patient underwent thoracoscopic surgery for esophageal carcinoma without complications. Conclusion: Esophageal carcinoma in patients with Crigler-Najjar syndrome is a rare case, and perioperative management is key in the treatment process. It is necessary to pay close attention to the changes of the disease to prevent complications.

Engineering of 177Lu-labeled gold encapsulated into dendrimeric nanomaterials for the treatment of lung cancer.

As a novel type of theranostic radioactive agents, 177Lu-labeled nanomaterials conjugated to macromolecules have been described. The study aimed to fabricate PAMAM-G4-(177Lu-dendrimer)-bombesin-folate in the dendrimeric cavity, assess the radiopharmaceutical ability for specifically targeted radiotherapy and simultaneously detects gastrin-releasing peptide receptors (GRPR) and folate receptors (FRs) overexpressed in lung carcinoma cells, respectively. In an aqueous-basic media, p-SCN-benzyl-DOTA was conjugated to the dendrimer. This dendrimer was formed by activating the carboxylic acid groups of DOTA-folic acid and bombesin with HATU and conjugating them to develop the dendrimer. As part of this process, the conjugate was combined with 1% HAuCl4, added NaBH4 and filtered by ultrafiltration. Infrared, UV-Vis, TEM analysis, dynamic light scattering (DLS), and fluorescence spectroscopy were employed to observe the composition of the fabricated sample. Radio-labeled 177LuCl3 was used to label the conjugate, which was then evaluated using the radio-HPLC method. Findings demonstrated dendrimeric functionalization with remarkable radiochemical composition purity up to >96%. Because of fluorescence studies, it was determined that the occurrence of AuNMs in the dendrimeric cavities gives beneficial photo-physical characteristics to the radiopharmaceutical for bio-imaging. HEL-299 lung cancer cells exhibited a selective absorption of the drug (%). It might be helpful as nuclear and optical imaging agents for lung cancers that overexpress FRs and GRPR and as a specific target for radiation therapy if combined with folate-bombesin.