Development of an Interpretable Deep Learning Model for Pathological Tumor Response Assessment After Neoadjuvant Therapy.

BACKGROUND: Neoadjuvant therapy followed by surgery has become the standard of care for locally advanced esophageal squamous cell carcinoma (ESCC) and accurate pathological response assessment is critical to assess the therapeutic efficacy. However, it can be laborious and inconsistency between different observers may occur. Hence, we aim to develop an interpretable deep-learning model for efficient pathological response assessment following neoadjuvant therapy in ESCC. METHODS: This retrospective study analyzed 337 ESCC resection specimens from 2020-2021 at the Pudong-Branch (Cohort 1) and 114 from 2021-2022 at the Puxi-Branch (External Cohort 2) of Fudan University Shanghai Cancer Center. Whole slide images (WSIs) from these two cohorts were generated using different scanning machines to test the ability of the model in handling color variations. Four pathologists independently assessed the pathological response. The senior pathologists annotated tumor beds and residual tumor percentages on WSIs to determine consensus labels. Furthermore, 1850 image patches were randomly extracted from Cohort 1 WSIs and binarily classified for tumor viability. A deep-learning model employing knowledge distillation was developed to automatically classify positive patches for each WSI and estimate the viable residual tumor percentages. Spatial heatmaps were output for model explanations and visualizations. RESULTS: The approach achieved high concordance with pathologist consensus, with an R^2 of 0.8437, a RAcc_0.1 of 0.7586, a RAcc_0.3 of 0.9885, which were comparable to two senior pathologists (R^2 of 0.9202/0.9619, RAcc_0.1 of 8506/0.9425, RAcc_0.3 of 1.000/1.000) and surpassing two junior pathologists (R^2 of 0.5592/0.5474, RAcc_0.1 of 0.5287/0.5287, RAcc_0.3 of 0.9080/0.9310). Visualizations enabled the localization of residual viable tumor to augment microscopic assessment. CONCLUSION: This work illustrates deep learning's potential for assisting pathological response assessment. Spatial heatmaps and patch examples provide intuitive explanations of model predictions, engendering clinical trust and adoption (Code and data will be available at https://github.com/WinnieLaugh/ESCC_Percentage once the paper has been conditionally accepted). Integrating interpretable computational pathology could help enhance the efficiency and consistency of tumor response assessment and empower precise oncology treatment decisions.

High spatial-resolved source-specific exposure and risk in the city scale: Influence of spatial interrelationship between PM2.5 sources and population on exposure.

Research on High Spatial-Resolved Source-Specific Exposure and Risk (HSRSSER) was conducted based on multiple-year, multiple-site synchronous measurement of PM2.5-bound (particulate matter with aerodynamic diameter<2.5 µm) toxic components in a Chinese megacity. The developed HSRSSER model combined the Positive Matrix Factorization (PMF) and Land Use Regression (LUR) to predict high spatial-resolved source contributions, and estimated the source-specific exposure and risk by personal activity time- and population-weighting. A total of 287 PM2.5 samples were collected at ten sites in 2018-2020, and toxic species including heavy metals (HMs), polycyclic aromatic hydrocarbons (PAHs) and organophosphate esters (OPEs) were analyzed. The percentage non-cancer risk were in the order of traffic emission (48 %) > industrial emission (22 %) > coal combustion (12 %) > waste incineration (11 %) > resuspend dust (7 %) > OPE-related products (0 %) ≈ secondary particles (0 %). Similar orders were observed in cancer risk. For traffic emission, due to its higher source contributions and large population in central area, non-cancer and cancer risk fraction increased from 23 % to 48 % and 20 % to 46 % after exposure estimation; while for industrial emission, higher source contributions but small population in suburb area decreased the percentage non-cancer and cancer risk from 38 % to 22 % and 39 % to 24 %, respectively.

Potential toxic components in size-resolved particles and gas from residential combustion: Emission factor and health risk.

Particulate matter (PM) from residential combustion is an existential threat to human health. Emission factors (EFs) of multiple potential toxic components (PTCs) in size-resolved PM and gas from eight residential fuel combustion were measured, and size distribution, gas/particle partitioning and health risks of the PTCs were investigated. Average EFs from clean coal and anthracite coal were PTEs (sum of EFs of 11 Potential Toxic Elements, 6.62 mg/kg fuels) > PAHs (sum of 22 Polycyclic Aromatic Hydrocarbons, 1.12 mg/kg) > OPAHs (sum of 5 Oxygenated Polycyclic Aromatic Hydrocarbons, 0.45 mg/kg) > PAEs (sum of 6 Phthalate Esters, 0.11 mg/kg) > NPAHs (sum of 14 Nitropolycyclic Aromatic Hydrocarbons, 16.84 µg/kg) > OPEs (sum of 7 Organophosphate Esters, 7.57 µg/kg) > PCBs (sum of 6 Polychorinated Biphenyls, 0.07 µg/kg), which were 2-3 and 1-2 orders of magnitude lower than the EFs of PTCs (except PTEs) from bituminous coal and biomass. Most PAHs, OPAHs and NPAHs, which may mainly originate from chemical reactions, showed similar size distributions and averagely 85 % concentrated in PM1. PTEs, PAEs, OPEs and PCBs generated from the release from raw fuels may have a higher proportion, so their size distributions were more complex and varied with combustion temperature, volatility of compounds, binding mode of the raw fuels, and so on. In addition, clean coal and high-quality anthracite coal could reduce the health risks from the potential organic toxic components, but also reveal the stumbling block of PTEs in risk control.

Evolutionary proteogenomic landscape from pre-invasive to invasive lung adenocarcinoma.

Lung adenocarcinoma follows a stepwise progression from pre-invasive to invasive. However, there remains a knowledge gap regarding molecular events from pre-invasive to invasive. Here, we conduct a comprehensive proteogenomic analysis comprising whole-exon sequencing, RNA sequencing, and proteomic and phosphoproteomic profiling on 98 pre-invasive and 99 invasive lung adenocarcinomas. The deletion of chr4q12 contributes to the progression from pre-invasive to invasive adenocarcinoma by downregulating SPATA18, thus suppressing mitophagy and promoting cell invasion. Proteomics reveals diverse enriched pathways in normal lung tissues and pre-invasive and invasive adenocarcinoma. Proteomic analyses identify three proteomic subtypes, which represent different stages of tumor progression. We also illustrate the molecular characterization of four immune clusters, including endothelial cells, B cells, DCs, and immune depression subtype. In conclusion, this comprehensive proteogenomic study characterizes the molecular architecture and hallmarks from pre-invasive to invasive lung adenocarcinoma, guiding the way to a deeper understanding of the tumorigenesis and progression of this disease.

DPA714 PET Imaging Shows That Inflammation of the Choroid Plexus Is Active in Chronic-Phase Intracerebral Hemorrhage.

PURPOSE: Our aims were to investigate the presence of choroid plexus (CP) inflammation in chronic-phase intracerebral hemorrhage (ICH) patients and to characterize any inflammatory cells in the CP. PATIENTS AND METHODS: An in vivo 18 F-DPA714 PET study was undertaken in 22 chronic-phase ICH patients who were admitted to the First Affiliated Hospital of Fujian Medical University or Tianjin Medical University General Hospital from April 2017 to June 2020. Ten control participants with nonhemorrhagic central nervous system diseases were included. Choroid plexus 18 F-DPA714 uptake was calculated as the average SUVR. To aid the interpretation of the 18 F-DPA714 uptake results at the CP level, Cy5-DPA714 in vivo imaging and immunofluorescence staining were used to show the presence of CP inflammation in an ICH mouse model during the chronic phase (14 weeks after ICH). Then immunofluorescence staining against translocator protein and other specific biomarkers was used to characterize the cells present in the inflamed CP of ICH mice in the chronic phase. RESULTS: PET imaging showed that CP DPA714 SUVRs in chronic-phase ICH patients were higher than in controls (mean CP SUVR ± SD; ICH group: 1.05 ± 0.35; control group: 0.81 ± 0.21; P = 0.006). Immunofluorescence staining of the CP in ICH model mice identified a population of CD45 + immune cells, peripheral monocyte-derived CD14 + cells, CD68 + phagocytes, and CD11b + resident microglia/macrophages expressing translocator protein, possibly contributing to the increased 18 F-DPA714 uptake. CONCLUSIONS: Our study shows that CP DPA714 uptake in chronic-phase ICH patients was higher than that of participants with nonhemorrhagic central nervous system diseases, which means that CP inflammation is still active in chronic-phase ICH patients.

Unlocking the Potential of Microwave Sterilization Technology in Ready-to-Eat Imitation Crab Meat Production.

Microwave sterilization is a novel potential sterilization technology to improve food quality. An industrial microwave sterilization system was used to sterilize imitation crab meat under thermal processing intensity F0 = 1, 2, 3. The characteristics of the microwave process, such as heating rate, processing time, and C100, were calculated. In addition, the quality of processed imitation crab meat was investigated. Compared with the conventional retort method, microwave sterilization significantly shortened the processing time of imitation crab meat by 63.71% to 72.45%. Under the same thermal processing intensity, microwave sterilization has demonstrated better results than retort sterilization in terms of water-holding capacity, color, and texture. Furthermore, microwave-treated imitation crab meat ingredients had a greater capacity to bind water molecules and obtained a more appropriate secondary protein structure. In addition, microwave technology can better preserve the unsaturated fatty acids (UFA) of imitation crab meat, which are 9.14%, 1.19%, and 0.32% higher than the traditional method at F0 = 1, 2, 3. The results would provide useful data for the subsequent research and development of ready-to-eat surimi products.

C1q+ tumor-associated macrophages contribute to immunosuppression through fatty acid metabolic reprogramming in malignant pleural effusion.

BACKGROUND: Although immune checkpoint blockade (ICB) therapy has shown remarkable benefits in cancers, a subset of patients with cancer exhibits unresponsiveness or develop acquired resistance due to the existence of abundant immunosuppressive cells. Tumor-associated macrophages (TAMs), as the dominant immunosuppressive population, impede the antitumor immune response; however, the underlying mechanisms have not been fully elucidated yet. METHODS: Single-cell RNA sequencing analysis was performed to portray macrophage landscape and revealed the underlying mechanism of component 1q (C1q)+ TAMs. Malignant pleural effusion (MPE) of human and mouse was used to explore the phenotypes and functions of C1q+ TAMs. RESULTS: C1q+ TAMs highly expressed multiple inhibitory molecules and their high infiltration was significantly correlated with poor prognosis. C1q+ TAMs promote MPE immunosuppression through impairing the antitumor effects of CD8+ T cells. Mechanistically, C1q+ TAMs enhance fatty acid binding protein 5 (FABP5)-mediated fatty acid metabolism, which activate transcription factor peroxisome proliferator-activated receptor-gamma, increasing the gene expression of inhibitory molecules. A high-fat diet increases the expression of inhibitory molecules in C1q+ TAMs and the immunosuppression of MPE microenvironment, whereas a low-fat diet ameliorates these effects. Moreover, FABP5 inhibition represses the expression of inhibitory molecules in TAMs and tumor progression, while enhancing the efficacy of ICB therapy in MPE and lung cancer. CONCLUSIONS: C1q+ TAMs impede antitumor effects of CD8+ T cells promoting MPE immunosuppression. Targeting C1q+ TAMs effectively alleviates the immunosuppression and enhances the efficacy of ICB therapy. C1q+ TAMs subset has great potential to be a therapeutic target for cancer immunotherapy.

Inhalation bioaccessibility and risk assessment for PM-bound organic components: Co-effects of component physicochemical properties, PM properties, and sources.

Inhalation bioaccessibility and deposition in respiratory tracts of organic components in atmospheric particulate matter (PM) are key factors for accurately estimating health risks and understanding human exposures. This study evaluated the in-vitro inhalation bioaccessibility of polycyclic aromatic hydrocarbons (PAHs) and PAH derivatives, phthalic acid esters (PAEs), polychlorinated biphenyls (PCBs), and organophosphate flame retardants (OPFRs) in size-resolved PM from a Chinese megacity. The bioaccessibility ranged from 0.2% to 77.8% in the heating period (HP), and from 0.7% to 94.2% in the non-heating period (NHP). Result suggests that less hydrophobic organics might be more bioaccessible. Bioaccessibility of medium logKow organics in sizes > 0.65 µm was significantly inhibited by high carbon fractions, indicating the co-effects. Then, this is the first study to explore effects of sources on inhalation bioaccessibility of organics. Coal and biomass combustion in HP and traffic emission in NHP negatively correlated with bioaccessibility. Secondary particles also negatively correlated with bioaccessibility of medium logKow organics. Incremental lifetime cancer risk (ILCR) and non-cancer risk (HQ) for all measured components in PM10 were estimated after considering the bioaccessibility and deposition efficiencies and the HQ and ILCR were within the acceptable range. BaP and DEHP were strong contributors to HQ and ILCR, respectively.

Functions and mechanisms of protein lysine butyrylation (Kbu): Therapeutic implications in human diseases.

Post-translational modifications (PTM) are covalent modifications of proteins or peptides caused by proteolytic cleavage or the attachment of moieties to one or more amino acids. PTMs play essential roles in biological function and regulation and have been linked with several diseases. Modifications of protein acylation (Kac), a type of PTM, are known to induce epigenetic regulatory processes that promote various diseases. Thus, an increasing number of studies focusing on acylation modifications are being undertaken. Butyrylation (Kbu) is a new acylation process found in animals and plants. Kbu has been recently linked to the onset and progression of several diseases, such as cancer, cardiovascular diseases, diabetes, and vascular dementia. Moreover, the mode of action of certain drugs used in the treatment of lymphoma and colon cancer is based on the regulation of butyrylation levels, suggesting that butyrylation may play a therapeutic role in these diseases. In addition, butyrylation is also commonly involved in rice gene expression and thus plays an important role in the growth, development, and metabolism of rice. The tools and analytical methods that could be utilized for the prediction and detection of lysine butyrylation have also been investigated. This study reviews the potential role of histone Kbu, as well as the mechanisms underlying this process. It also summarizes various enzymes and analytical methods associated with Kbu, with the goal of providing new insights into the role of Kbu in gene regulation and diseases.

Analysis of quality differences between Scutellaria baicalensis Georgi and Scutellaria rehderiana Diels based on phytochemistry and bioactivity evaluation.

Scutellaria baicalensis Georgi (SG) and Scutellaria rehderiana Diels (SD) belong to the same genus of Scutellaria in the Labiatae (Lamiaceae) family. SG is confirmed as the medicinal source according to the Chinese Pharmacopeia, but SD is often used as a substitute for SG due to its abundant plant resources. However, the current quality standards are far from sufficient to judge the quality differences between SG and SD. In this study, an integrated strategy of "biosynthetic pathway (specificity) - plant metabolomics (difference) - bioactivity evaluation (effectiveness)" was established to evaluate this quality differences. First, an ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS/MS) method was developed for the identification of chemical components. The abundant components information was obtained and the characteristic constituents were screened according to the location in the biosynthetic pathway as well as species specificity. Then, plant metabolomics combined with multivariate statistical analysis to find differential components between SG and SD. The chemical markers for quality analysis were determined based on the differential and characteristic components, and the content of each marker was tentatively evaluated through the semi-quantitative analysis of UHPLC-Q/TOF-MS/MS. Finally, the anti-inflammatory activity of SG and SD was compared by measuring the inhibitory effect on the release of NO from lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Under this analytical strategy, a total of 113 compounds were tentatively identified in both SG and SD, among which baicalein, wogonin, chrysin, oroxylin A 7-O-ß-D-glucuronoside, pinocembrin and baicalin were selected as chemical markers due to their species characteristics and differentiation. The contents of oroxylin A 7-O-ß-D-glucuronoside and baicalin was higher in SG, and the others were higher in SD. In addition, both SG and SD exhibited prominent anti-inflammatory activity, but SD was less effective. The analysis strategy combining phytochemistry and bioactivity evaluation realized the scientific evaluation of the intrinsic quality differences between SG and SD, which provides a reference for fully utilizing and expanding the medicinal resources, and also provides a reference for the comprehensive quality control of herbal medicines.

Analysis of the dust-methane two-phase coupling blowdown effect at different air duct positions in an excavation anchor synchronous tunnel.

Bolter miners are being increasingly used. Unfortunately, this mining technology causes a considerable amount of air pollution (especially by methane and dust) during excavation. In this study, the multiphase coupling field of airflow-dust-methane for different distances between the pressure air outlet and the working face (Lp) was simulated by using the FLUENT software. The migration law of pollutants in the multiphase coupling field was analyzed, and the distance parameters between the pressure air outlet and the working face were optimized. Finally, the simulation results were verified based on the field measurement results. We found that the blowdown effect was more obvious when 14 m ≤ Lp < 16 m compared with other conditions. The peak value of dust concentration within this distance range was the smallest (44.4% lower than the highest peak value, which was verified when Lp = 18 m), while the methane concentration was < 0.6%. A high-concentration area (where methane concentration > 0.75%), identified near the walking part of the bolter miner, was 13 m shorter than the largest (when Lp = 18 m). Therefore, we determined that the optimal blowdown distance would be 14 m ≤ Lp < 16 m. Within this range, the dust removal and methane dilution effects are optimal, effectively improving the tunnel air quality and providing a safe and clean environment for mine workers.

POU2F3: A Sensitive and Specific Diagnostic Marker for Neuroendocrine-low/negative Small Cell Lung Cancer.

POU2F3 (POU class 2 homeobox 3) is a novel transcription factor used to define the special molecular subtype of small cell lung cancer (SCLC) known as SCLC-P. Nevertheless, the sensitivity and specificity of POU2F3 immunohistochemical (IHC) staining have not been fully investigated. In this study, we explored the expression of POU2F3 by IHC in a large cohort of SCLC clinical samples (n=246), other common lung cancer types (n=2207), and various other cancer types (n=194). The results showed that POU2F3 was strongly nuclear stained in 13.41% (33/246) of SCLC cases, with negative or minimal labeling for thyroid transcription factor-1 and neuroendocrine (NE) markers. Compared with POU2F3-negative SCLC, SCLC-P harbored fewer TP53 and RB1 mutations. POU2F3 was also expressed in 3.13% (8/256) of squamous cell carcinomas (SCCs) and 20% (2/10) of large cell NE carcinomas (LCNECs), whereas other lung cancer types were negative. In addition to lung cancer, POU2F3 was positive in 22.2% (4/18) of thymic tumors. All other tumors were POU2F3-negative except for thymic carcinoma, although sparsely distributed weak nuclear staining was observed in lung adenocarcinoma, cervical SCC, and colorectal carcinoma. The sensitivity and specificity of POU2F3 in NE-low/negative SCLC were 82.1% and 99.4%, respectively. Notably, some rare unique patterns of POU2F3 expression were observed. One case of thymic SCC was characterized by diffuse and uniform cytomembrane staining. One case of esophageal NE tumor was nuclear-positive, while the normal proliferating squamous epithelium was strongly membrane-stained. This is the largest cohort of clinical samples to confirm that POU2F3 is a highly sensitive and specific diagnostic marker for NE-low/negative SCLC.

[Health Impacts of Air Pollution in Tianjin].

Ambient air pollution is a dominant determinant of health. The health effects and economic losses due to air pollution are very important for decision-making. Since the implementation of the "Air Pollution Prevention and Control Action Plan" and "blue sky defense war" policies, the air quality of Tianjin has changed significantly. Here, the health effects and economic losses attributable to ambient air pollution in Tianjin from 2013 to 2020 wereestimated. For the particulate matter which has complex components, we assessed the inhalation health risks of heavy metals and polycyclic aromatic hydrocarbons (PAHs) in PM2.5. The variation in the concentration of the main components of PM2.5 was also analyzed. The results showed that improved air quality had positive health benefits. The health benefits from SO2 were the highest among the six air pollutants, and 3786 deaths were avoided in 2020 compared to in 2013 due to lower SO2 concentration. The economic losses caused by air pollutants ranged from several billion to ten billion yuan. Among the six air pollutants, particulate matter and ozone had higher health losses in recent years. The health risks of heavy metals and PAHs in PM2.5 showed a decreasing trend. However, Cr(Ⅵ), As, Cd, and Ni in PM2.5in the winter of 2020 still had respiratorysystem carcinogenic risk, whereas there was no health risk of PAHs in PM2.5in 2019-2020. The concentrations of main components of PM2.5 have decreased significantly. In the future, the reduction of health loss caused by air pollution depends on synergy governance of particulate matter and ozone and further research on health effects.

Promising immunotherapeutic targets in lung cancer based on single-cell RNA sequencing.

Immunotherapy has made great strides in the treatment of lung cancer, but a significant proportion of patients still do not respond to treatment. Therefore, the identification of novel targets is crucial to improving the response to immunotherapy. The tumor microenvironment (TME) is a complex niche composed of diverse pro-tumor molecules and cell populations, making the function and mechanism of a unique cell subset difficult to understand. However, the advent of single-cell RNA sequencing (scRNA-seq) technology has made it possible to identify cellular markers and understand their potential functions and mechanisms in the TME. In this review, we highlight recent advances emerging from scRNA-seq studies in lung cancer, with a particular focus on stromal cells. We elucidate the cellular developmental trajectory, phenotypic remodeling, and cell interactions during tumor progression. Our review proposes predictive biomarkers and novel targets for lung cancer immunotherapy based on cellular markers identified through scRNA-seq. The identification of novel targets could help improve the response to immunotherapy. The use of scRNA-seq technology could provide new strategies to understand the TME and develop personalized immunotherapy for lung cancer patients.

miR-132-3p promotes heat stimulation-induced esophageal squamous cell carcinoma tumorigenesis by targeting KCNK2.

Epidemiological evidence supports that consumption of high-temperature food and beverages is an important risk factor for esophageal squamous cell carcinoma (ESCC); however, the underlying mechanism still remains unclear. Here, we established a series of animal models and found that drinking 65°C water can promote esophageal tumor progression from preneoplastic lesions to ESCC. RNA sequencing data showed that miR-132-3p was highly expressed in the heat stimulation group compared with controls. Further study verified that miR-132-3p were upregulated in human premalignant lesion tissues of the esophagus, ESCC tissues, and cells. Overexpression of miR-132-3p could promote ESCC cell proliferation and colony formation, whereas knockdown of miR-132-3p could inhibit ESCC progression in vitro and in vivo. Importantly, dual-luciferase reporter assays showed that miR-132-3p could bind with the 3'-untranslated region of KCNK2 and inhibit KCNK2 gene expression. Knockdown or overexpression of KCNK2 could promote or suppress ESCC progression in vitro. These data suggest that heat stimulation can promote ESCC progression and miR-132-3p mediated this process by directly targeting KCNK2.

Variations of inhalation risks during different heavy pollution episodes based on 3-year measurement of toxic components in size-segregated particles.

Toxic metals (TMs) and polycyclic aromatic hydrocarbons (PAHs) in size-segregated particles during common days (CD) and different heavy pollution (HP) episodes were measured during 2018-2021 in a Chinese megacity. The Multiple Path Particle Dosimetry Model (MPPD) was performed to estimate deposition efficiency, and then inhalation risks in the human pulmonary region during different types of HP were assessed and compared. The higher pulmonary deposition efficiency of PAHs and TMs during all types of HP than those during CD was confirmed. The accumulative incremental lifetime cancer risk (ILCR) of different HP were 2.42 × 10-5, 1.52 × 10-5, 1.39 × 10-5, 1.30 × 10-5 and 2.94 × 10-6 for HP4 (combustion sources HP), HP1 (ammonium nitrate HP), HP5 (mixed sources HP), HP3 (resuspended dust HP) and HP2 (ammonium sulfate HP), respectively. The accumulative hazard quotient (HQ) during different HP episodes decreased in the order of HP4 (0.32) > HP3 (0.24) > HP1 (0.22) > HP5 (0.18) > HP2 (0.05). The inhalation risks were dominated by Ni and Cr, what's more, the HQ of Ni and ILCR of Cr during the five HP episodes shared a similar size distribution pattern. However, the characteristic components during different HP episodes and their size distributions of them were distinctive. The size distribution of inhalation risks of the related components (Ni, Cr, BaP, and As) from the combustion process during HP4 peaked at fine mode (0.65-2.1 µm). The size distribution of inhalation risks of the dust-related components (Mn and V) and the components (As and BaP) that are likely to volatilize and re-distribution peaked at coarse mode (2.1-3.3 µm) during HP3. Notably, Mn and Co as catalysts at fine mode could increase the degree of secondary formation and toxicity.

Prognostic value of tumor immune microenvironment factors in patients with stage I lung adenocarcinoma.

Survival is difficult to predict in patients with resected stage I lung adenocarcinoma (LUAD), but tumor microenvironment (TME) factors appear useful in predicting survival in advanced non-small cell lung cancer. We aimed to identify the TME factors linked to recurrence/metastasis and survival in stage I LUAD patients. We evaluated TME factors in stage I LUAD patients in The Cancer Genome Atlas (TCGA) using the "ESTIMATE" and "MCP-counter" R packages. We characterized infiltrating immune cells in the tumor and stromal regions in 44 stage I LUAD patients at our hospital using immunohistochemical methods combined with the HALO® Image Analysis Platform. In TCGA LUAD patients, the number of neutrophils was higher in patients without recurrence/metastasis than in patients with recurrence/metastasis. For patients with recurrence/metastasis, higher CD8+ T lymphocyte and B lymphocyte infiltration levels were associated with better overall survival (OS), and myeloid dendritic cell (DC) infiltration was associated with better disease-free survival (DFS). In stage I LUAD patients at our hospital, CD4+ T cells, CD8+ T cells, CD14+ monocytic lineage cells, CD16+ NK cells, and CD19+ B lymphocytes were more highly expressed in stromal regions than in tumor regions. Moreover, high intratumoral CD11c+ myeloid DC and CD68+ macrophage levels were associated with recurrence/metastasis. Within tumor regions, higher CD11c+ myeloid DC and CD68+ macrophage levels were associated with shorter DFS; within stromal regions, higher CD68+ macrophage levels were associated with shorter DFS. Multivariate analysis revealed that the presence of intravascular carcinoma embolus, higher intratumoral CD11c+ myeloid DC levels, and high stromal CD68+ macrophage and CD4+ T-cell levels were independently linked to recurrence/metastasis in stage I LUAD patients. This study using 2 datasets shows that key players in the TME are associated with recurrence/metastasis in stage I LUAD patients. Higher intratumoral CD11c+ myeloid DC, stromal CD68+ macrophage and stromal CD4+ T-cell levels are independent prognostic factors for DFS in these patients.

Protective effect and mechanism of ginsenoside Rg2 on atherosclerosis.

Background: Ginsenoside Rg2 (Rg2) has a variety of pharmacological activities and provides benefits during inflammation, cancer, and other diseases. However, there are no reports about the relationship between Rg2 and atherosclerosis. Methods: We used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to detect the cell viability of Rg2 in vascular smooth muscle cells (VSMCs) and human umbilical vein endothelial cells (HUVECs). The expression of inflammatory factors in HUVECs and the expression of phenotypic transformation-related marker in VSMCs were detected at mRNA levels. Western blot method was used to detect the expression of inflammation pathways and the expression of phenotypic transformation at the protein levels. The rat carotid balloon injury model was performed to explore the effect of Rg2 on inflammation and phenotypic transformation in vivo. Results: Rg2 decreased the expression of inflammatory factors induced by lipopolysaccharide in HUVECs-without affecting cell viability. These events depend on the blocking regulation of NF-κB and p-ERK signaling pathway. In VSMCs, Rg2 can inhibit the proliferation, migration, and phenotypic transformation of VSMCs induced by platelet derived growth factor-BB (PDGF-BB)-which may contribute to its anti-atherosclerotic role. In rats with carotid balloon injury, Rg2 can reduce intimal proliferation after injury, regulate the inflammatory pathway to reduce inflammatory response, and also suppress the phenotypic transformation of VSMCs. Conclusion: These results suggest that Rg2 can exert its anti-atherosclerotic effect at the cellular level and animal level, which provides a more sufficient basis for ginseng as a functional dietary regulator.

Frequent EGFR exon 20 insertion in the so-called peripheral-type squamous cell neoplasm of uncertain malignant potential: a variant of bronchiolar adenoma or under-recognised entity?

INTRODUCTION: Herein we describe a series of rare peripheral pulmonary neoplasms temporarily termed "peripheral type squamous cell neoplasm of uncertain malignant potential (PSCN-UMP)" and investigate their relationship to bronchiolar adenoma (BA) and squamous cell carcinoma (SCC). MATERIALS AND METHODS: The histologic and immunohistochemical features of 10 PSCN-UMPs and six BAs were compared. Whole exome sequencing (WES) and bioinformatics analysis were performed to further compare the genetic features of PSCN-UMPs, BAs, and NSCLCs. RESULTS: All PSCN-UMPs were peripherally located and histologically characterised by the lepidic, nested, and papillary proliferation of relatively bland squamous cells, accompanied by entrapped hyperplastic reactive pneumocytes. The basal squamous cells coexpressed TTF1 and squamous markers. Both cellular components exhibited bland morphology and a low proliferative activity. The six BAs met the morphologic and immunophenotypic features of proximal-type BA. Genetically, driver mutations, including frequent EGFR exon 20 insertions, were found in PSCN-UMPs, while the KRAS mutation, BRAF mutation, and ERC1::RET fusion were detected in BAs. PSCN-UMPs also shared some alterations with BAs in mutational signatures, while copy number variants (CNV) were enriched in MET and NKX2-1 in PSCN-UMP and MCL1, MECOM, SGK1, and PRKAR1A in BA. CONCLUSION: PSCN-UMPs exhibited the proliferation of bland squamous cells accompanied by entrapped pneumocytes and frequent EGFR exon 20 insertions, which showed distinct features from BAs and SCCs. Recognition of this specific entity will help to expand the morphologic and molecular spectrum of peripheral lung squamous neoplasms.

Effects of cooling rates during depuration on the quality of Pacific oysters (Crassostrea gigas) at anhydrous preservation stage.

The Pacific oyster could be affected by several pressure sources during cold chain logistics, which reduce the quality of oyster, and even improve its mortality. For improving the quality of oyster, the effects of depuration process at different cooling rates (1, 3, 7, 11 and 16 °C/h) on selected Pacific oyster were studied. The results indicated that extreme fluctuations in the depuration temperatures could affect the survival rates and qualities of oysters. The oysters exhibited low survival rates, glycogen contents and pH values at an increased cooling rate. Their contents in the 1 °C/h group after 3 d preservation were 100 %, 16.30 ± 1.64 mg/100 g and 6.72 ± 0.01, respectively, while there were 71 %, 7.72 ± 0.88 mg/100 g and 6.53 ± 0.01 in the 16 °C/h group after 3 d preservation, respectively. Furthermore, the ATP-related compounds were affected by the different cooling rates. AMP and IMP were the main ATP-related compounds, and their contents in the 1 °C/h group after 3 d preservation were 37.21 ± 1.10 mg/100 g and 29.47 ± 1.10 mg/100 g, respectively, while there were 32.07 ± 1.10 mg/100 g and 13.16 ± 1.60 mg/100 g in the 16 °C/h group after 3 d preservation, respectively. The proportions of the total umami, as well as the sweet amino acids also decreased, the proportions of the umami amino acids and sweet amino acids in the total amino acids, were 31.37%-38.80%, and their proportions in 1 °C/h group were higher than that in 16 °C/h group. Conversely, the fatty acid content of each group exhibited significant differences. Combined with the above results, the oyster maintained a high survival rate and higher quality at a cooling rate of 1 °C/h during depuration.