Global analysis of T-cell groups reveals immunological features and common antigen targets of digestive tract tumors.

BACKGROUND: T cells are key players in the tumor immune microenvironment (TIME), as they can recognize and eliminate cancer cells that express neoantigens derived from somatic mutations. However, the diversity and specificity of T-cell receptors (TCRs) that recognize neoantigens are largely unknown, due to the high variability of TCR sequences among individuals. METHODS: To address this challenge, we applied GLIPH2, a novel algorithm that groups TCRs based on their predicted antigen specificity and HLA restriction, to cluster the TCR repertoire of 1,702 patients with digestive tract cancer. The patients were divided into five groups based on whether they carried tumor-infiltrating or clonal-expanded TCRs and calculated their TCR diversity. The prognosis, tumor subtype, gene mutation, gene expression, and immune microenvironment of these groups were compared. Viral specificity inference and immunotherapy relevance analysis performed for the TCR groups. RESULTS: This approach reduced the complexity of TCR sequences to 249 clonally expanded and 150 tumor-infiltrating TCR groups, which revealed distinct patterns of TRBV usage, HLA association, and TCR diversity. In gastric adenocarcinoma (STAD), patients with tumor-infiltrating TCRs (Patients-TI) had significantly worse prognosis than other patients (Patients-nonTI). Patients-TI had richer CD8+ T cells in the immune microenvironment, and their gene expression features were positively correlated with immunotherapy response. We also found that tumor-infiltrating TCR groups were associated with four distinct tumor subtypes, 26 common gene mutations, and 39 gene expression signatures. We discovered that tumor-infiltrating TCRs had cross-reactivity with viral antigens, indicating a possible link between viral infections and tumor immunity. CONCLUSION: By applying GLIPH2 to TCR sequences from digestive tract tumors, we uncovered novel insights into the tumor immune landscape and identified potential candidates for shared TCRs and neoantigens.

Preoperative C-reactive Protein and Other Inflammatory Biomarkers as Predictors of Postoperative Complications in Colorectal Tumor Patients.

Objective: This study aims to explore the predictive value of preoperative C-reactive protein (CRP) and other inflammatory biomarkers: platelet-to-lymphocyte ratio (PLR) and neutrophil-to-lymphocyte ratio (NLR) for postoperative complications (infection, diarrhea, etc.) in colorectal tumor patients. Methods: A cohort of 109 colorectal tumor patients who underwent surgical resection for colorectal cancer at the hospital from January 2021 to December 2022 were selected as the research subjects. Patients' postoperative complications were tracked, and they were split into the complication group and the non-complication group. All patients underwent preoperative biochemical tests. Serum levels of CRP, PLR, and NLR were compared between the two groups. The relationship between these markers and postoperative complications in colorectal tumor patients was analyzed. A Logistic regression model was established to analyze their impact on postoperative complications, and a Receiver Operating Characteristic (ROC) curve was drawn to assess predictive value. Results: Among the 109 colorectal tumor patients, 31 cases (28.44%) experienced postoperative complications. The complication group had larger tumor diameters and a higher proportion of open surgeries compared to the non-complication group (P < .05). Serum levels of CRP, PLR, and NLR were higher in the complication group compared to the non-complication group (P < .05). Correlation analysis showed that serum CRP, PLR, and NLR values were positively correlated with postoperative complications in colorectal tumor patients (r > 0, P < .05). The Logistic regression model revealed that high serum CRP levels (95%CI: 1.253-2.503), PLR (95%CI: 1.005-1.041), and NLR values (95%CI: 2.702-20.533) were risk factors for postoperative complications in colorectal tumor patients (OR>1, P < .05). The ROC curve demonstrated that serum CRP levels, PLR, and NLR values had certain predictive values for postoperative complications in colorectal tumor patients (AUC=0.811, 0.789, 0.870), the optimal predictive values were obtained when the cut-off values were set at 5.400 mg/L, 142.790, and 2.485, respectively and combined detection showed even higher predictive values (AUC=0.913). At 1 week post-surgery, the patient's CRP levels, PLR value, and NLR value were significantly lower than pre-surgery (P < .05). Conclusion: Preoperative serum CRP, PLR, and NLR values are closely related to postoperative complications in colorectal tumor patients, and they can be used to predict the risk of postoperative complications in colorectal tumor patients. Clinically, early prediction of postoperative complications in patients can be achieved by measuring the aforementioned indicators, allowing for the implementation of appropriate preventive measures such as detoxification and infection control to improve patient outcomes.

Polystyrene nanoplastics-induced lung apoptosis and ferroptosis via ROS-dependent endoplasmic reticulum stress.

It has been shown that exposure to nanoplastics (MNPs) through inhalation can induce pulmonary toxicity, but the toxicological mechanism of MNPs on the respiratory system remains unclear. Therefore, we explored the toxicological mechanism of exposure to polystyrene nanoplastics (PS-NPs) (0.05, 0.15, 0.2 mg/mL) on BEAS-2B cells. Results revealed that PS-NPs induce oxidative stress, increased apoptosis rate measured by flow cytometry, the key ferroptosis protein (GPX4 and FTH1) reduction, increased iron content, mitochondrial alterations, and increased malondialdehyde (MDA) level. Besides, consistent results were observed in mice exposed to PS-NPs (5 mg/kg/2d, 10 mg/kg/2d). Thus, we proved that PS-NPs induced cell death and lung damage through apoptosis and ferroptosis. In terms of mechanism, the elevation of the endoplasmic reticulum (ER) stress protein expression (IRE1α, PERK, XBP1S, and CHOP) revealed that PS-NPs induce lung damage by activating the two main ER stress pathways. Furthermore, the toxicological effects of PS-NPs observed in this study are attenuated by the ROS inhibitor N-acetylcysteine (NAC). Collectively, NPs-induced apoptosis and ferroptosis are attenuated by NAC via inhibiting the ROS-dependent ER stress in vitro and in vivo. This improves our understanding of the mechanism by which PS-NPs exposure leads to pulmonary injury and the potential protective effects of NAC.

Sevoflurane suppresses colorectal cancer malignancy by modulating ß-catenin ubiquitination degradation via circSKA3.

BACKGROUND: Sevoflurane (SEV), a commonly used inhalational anesthetic, reportedly inhibits colorectal cancer (CRC) malignancy, but whether SEV can inhibit the malignancy of CRC by regulating circular RNAs (circRNAs) remains unclear. Therefore, we aimed to identify specific circRNAs that may be affected by SEV and to investigate their functional roles in CRC. METHODS: RT-qPCR was employed to detect the expression of circRNAs and mRNAs in CRC cells and tissues. Fluorescence in situ hybridization (FISH) was used to determine the location of circSKA3. Protein expression was assessed by western blot analysis. Function-based in vitro and in vivo experiments, including CCK-8, colony formation, transwell, and apoptosis assays and mouse xenograft tumor models, were conducted using circSKA3-knockdown and circSKA3-overexpression cell lines. RNA immunoprecipitation, RNA pull-down and mass spectrometry analyses were performed to explore the related mechanism. RESULTS: Our findings revealed that SEV could inhibit CRC cell activity, proliferation and migration and promote apoptosis in CRC cells. We found that circSKA3 was upregulated in CRC and associated with poorer survival and that its expression could be reduced by SEV. The overexpression of circSKA3 reversed the effects of SEV on inhibiting cell activity, proliferation and migration and promoting apoptosis. The mechanistic analysis revealed that circSKA3 could bind to the ARM structural domain of ß-catenin and thereby disrupt its interaction with the CK1/GSK3ß/ß-TrCP1 destruction complex, resulting in the ubiquitinated degradation of ß-catenin and the activation of Wnt/ß-catenin signaling. In addition, SEV downregulated circSKA3 in vivo to inhibit tumor growth. CONCLUSIONS: All the results showed that SEV could inhibit CRC progression via circSKA3 by increasing ß-catenin ubiquitination degradation.

Spatiotemporal Quantification of HER2-targeting Antibody-Drug Conjugate Bystander Activity and Enhancement of Solid Tumor Penetration.

PURPOSE: Antibody-drug conjugate (ADC) has had a transformative effect on the treatment of many solid tumors, yet it remains unclear how ADCs exert bystander activity in the tumor microenvironment. EXPERIMENTAL DESIGN: Here, we directly visualized and spatiotemporally quantified the intratumor biodistribution and pharmacokinetics of different ADC components by developing dual-labeled fluorescent probes. RESULTS: Mechanistically, we found that tumor penetration of ADCs is distinctly affected by their ability to breach the binding site barrier (BSB) in perivascular regions of tumor vasculature, and bystander activity of ADC can only partially breach BSB. Furthermore, bystander activity of ADCs can work in synergy with coadministration of their parental antibodies, leading to fully bypassing BSBs and enhancing tumor penetration via a two-step process. CONCLUSIONS: These promising preclinical data allowed us to initiate a phase I/II clinical study of coadministration of RC48 and trastuzumab in patients with malignant stomach cancer to further evaluate this treatment strategy in humans.

Peptostreptococcus anaerobius is a potential diagnostic biomarker of colorectal cancer.

Accumulating evidences have shown that Peptostreptococcus anaerobius (P.a) is abundantly enriched in the fetus of colorectal cancer (CRC) patients. P.a is reported able to invade colorectal tissues. This study intends to uncover the clinical significance of P.a in CRC. Mucosal tissues collected from CRC cases (n=109) and precancerous healthy ones (n=65) were subjected to the determination of the absolute copy number of P.a by droplet digital PCR. The positive rate of P.a in mucosal tissues of CRC and healthy ones was 79.8% (87/109), and 55.4% (36/65), respectively. The average absolute copy number of P.a in them was 2.3 copy/ng DNA, and 0.32 copy/ng DNA, respectively. The abundance of P.a in mucosal tissues of CRC, and age and TNM staging of CRC cases were correlated to its survival. The abundance of P.a in CRC cases was remarkably correlated to the relative level of SQLE. The abundance of P.a can be monitored to predict the prognosis of CRC.

Protocol for PPP1R15A-inhibited mouse model establishment with subcutaneous B16F1 tumor and single-cell analysis.

Here, we present a protocol for exploring the effects of PPP1R15A inhibitor, Sephin1, on antitumor immunity of B16F1 subcutaneous tumor in mice. We describe steps for constructing single-cell transcriptome and TCR libraries, sequencing, and using sequencing data for the integration of expression and TCR data. We then detail procedures for gene differentiation, regulon and cell-cell communication analysis, and validation of single-cell analysis results. For complete details on the use and execution of this protocol, please refer to Wang et al.1.

Small molecule activators of TAK1 promotes its activity-dependent ubiquitination and TRAIL-mediated tumor cell death.

TAK1 is a key modulator of both NF-κB signaling and RIPK1. In TNF signaling pathway, activation of TAK1 directly mediates the phosphorylation of IKK complex and RIPK1. In a search for small molecule activators of RIPK1-mediated necroptosis, we found R406/R788, two small molecule analogs that could promote sustained activation of TAK1. Treatment with R406 sensitized cells to TNF-mediated necroptosis and RIPK1-dependent apoptosis by promoting sustained RIPK1 activation. Using click chemistry and multiple biochemical binding assays, we showed that treatment with R406 promotes the activation of TAK1 by directly binding to TAK1, independent of its original target Syk kinase. Treatment with R406 promoted the ubiquitination of TAK1 and the interaction of activated TAK1 with ubiquitinated RIPK1. Finally, we showed that R406/R788 could promote the cancer-killing activities of TRAIL in vitro and in mouse models. Our studies demonstrate the possibility of developing small molecule TAK1 activators to potentiate the effect of TRAIL as anticancer therapies.

Analysis of cumulative live birth rate outcomes of three ovarian stimulation protocols in patients after laparoscopic cystectomy of ovarial endometrioma: a retrospective cohort study.

BACKGROUND: Previous studies have reported that after laparoscopic cystectomy of ovarial endometrioma, the ovarian response to gonadotropin (Gn) significantly decreased. However, for patients with diminished ovarian reserve (DOR) after ovarian surgery, how to choose the most appropriate controlled ovarian hyperstimulation protocol has not been concluded. Compared with the traditional agonist regimen, the gonadotropin (Gn)-releasing hormone (GnRH) antagonist, microstimulation, and progestin-primed ovarian stimulation (PPOS) protocols are simple to operate and have a shorter cycle, which are often used in patients with DOR. So the purpose of our study is to compare the assisted reproductive outcomes of these three controlled ovarian hyperstimulation protocols in patients with DOR following laparoscopic cystectomy of ovarial endometrioma. METHODS: In this retrospective cohort study, 89 patients with DOR who had undergone in vitro fertilisation/intracytoplasmic sperm injection at the Department of Reproductive Medicine at the Third Affiliated Hospital of Zhengzhou University from 1 to 2018 to 31 December 2020 were included. According to the controlled ovarian hyperstimulation protocols employed, the patients were divided into GnRH antagonist (38 patients), PPOS (27 patients), and microstimulation (24 patients) groups. The basic data and clinical outcomes of the three groups were compared. The main outcome measure was the cumulative live birth rate. RESULTS: No significant differences in the age of the female patients and their spouses and female patients' body mass index and basal endocrine levels (follicle-stimulating hormone and oestradiol) were noted among the three groups (P > 0.05). The GnRH antagonist group had higher antral follicle counts, greater endometrial thickness on the human chorionic Gn injection day, greater number of oocytes retrieved, and higher two pronuclear embryo counts than did the other two groups. However, the starting dosage of Gn was lower in the GnRH antagonist group than in the other two groups. The microstimulation group had a significantly higher oocyte output rate and high-quality embryo rate than did the other two groups (P < 0.05). No significant differences in the total dosage of Gn, cumulative pregnancy rate, cumulative live birth rate, viable embryo rate, and blastocyst formation rate were observed among the three groups (P > 0.05). CONCLUSION: In conclusion, for patients aged under 40 years who experienced DOR after laparoscopic cystectomy of ovarial endometrioma, GnRH antagonist protocol and PPOS protocol can obtain better ovulation induction outcomes and cumulative live birth rate than microstimulation protocol, and are more suitable ovulation induction protocols.

Stimuli-Responsive PROTACs for Controlled Protein Degradation.

Proteolysis Targeting Chimeras (PROTACs) represent a promising therapeutic modality to address undruggable and resistant issues in drug discovery. However, potential on-target toxicity remains clinically challenging. We developed a generalized caging strategy to synthesize a series of stimuli-responsive PROTACs (sr-PROTACs) with diverse molecular blocks bearing robust and cleavable linkers, presenting "turn on" features in manipulating protein degradation. By leveraging pathological cues, such as elevated ROS, phosphatase, H2 S, or hypoxia, and external triggers, such as ultraviolet light, X-Ray, or bioorthogonal reagents, we achieved site-specific activation and traceless release of original PROTACs through de-caging and subsequent self-immolative cleavage, realizing selective uptake and controlled protein degradation in vitro. An in vivo study revealed that two sr-PROTACs with phosphate- and fluorine-containing cages exhibited high solubility and long plasma exposure, which were specifically activated by tumor overexpressing phosphatase or low dosage of X-Ray irradiation in situ, leading to efficient protein degradation and potent tumor remission. With more reactive biomarkers to be screened from clinical practice, our caging library could provide a general tool to design activatable PROTACs, prodrugs, antibody-drug conjugates, and smart biomaterials for personalized treatment, tissue engineering or regenerative medicine.

Mitochondrial Protease Targeting Chimeras for Mitochondrial Matrix Protein Degradation.

Targeted protein degradation (TPD) is an emerging technique for protein regulation. Currently, all TPD developed in eukaryotic cells relies on either ubiquitin-proteasome or lysosomal systems, thus are powerless against target proteins in membrane organelles lacking proteasomes and lysosomes, such as mitochondria. Here, we developed a mitochondrial protease targeting chimera (MtPTAC) to address this issue. MtPTAC is a bifunctional small molecule that can bind to mitochondrial caseinolytic protease P (ClpP) at one end and target protein at the other. Mechanistically, MtPTAC activates the hydrolase activity of ClpP while simultaneously bringing target proteins into proximity with ClpP. Taking mitochondrial RNA polymerase (POLRMT) as a model protein, we have demonstrated the powerful proteolytic ability and antitumor application prospects of MtPTAC, both in vivo and in vitro. This is the first modularly designed TPD that can specifically hydrolyze target proteins inside mitochondria.

Single-cell RNA sequencing reveals the suppressive effect of PPP1R15A inhibitor Sephin1 in antitumor immunity.

Protein phosphatase 1 regulatory subunit 15A (PPP1R15A) is an important factor in the integrated stress response (ISR) in mammals and may play a crucial role in tumorigenesis. In our studies, we found an inhibitor of PPP1R15A, Sephin1, plays a protumorigenic role in mouse tumor models. By analyzing the single-cell transcriptome data of the mouse tumor models, we found that in C57BL/6 mice, Sephin1 treatment could lead to higher levels of ISR activity and lower levels of antitumor immune activities. Specifically, Sephin1 treatment caused reductions in antitumor immune cell types and lower expression levels of cytotoxicity-related genes. In addition, T cell receptor (TCR) repertoire analysis demonstrated that the clonal expansion of tumor-specific T cells was inhibited by Sephin1. A special TCR + macrophage subtype in tumor was identified to be significantly depleted upon Sephin1 treatment, implying its key antitumor role. These results suggest that PPP1R15A has the potential to be an effective target for tumor therapy.

Longitudinal Immune Profiling Highlights CD4+ T Cell Exhaustion Correlated with Liver Fibrosis in Schistosoma japonicum Infection.

Schistosomiasis remains an important public health concern. The eggs deposited in livers invoke a Th2-dominant response, which mediates the fibrotic granulomatous response. However, the mechanisms involved in this immunopathological process are still not perfectly clear. Here, we report a single-cell transcriptional landscape of longitudinally collected BALB/c mouse splenocytes at different time points after Schistosoma japonicum infection. We found that exhausted CD4+ T cells were enriched after infection, changing from coproducing multiple cytokines to predominantly producing the Th2 cytokine IL-4. Regulatory B cells had high expression of Fcrl5, Ptpn22, and Lgals1, potentially regulating exhausted CD4+ T cells via direct PD-1-PD-L2 and PD-1-PD-L1 interactions. Within the myeloid compartment, the number of precursor and immature neutrophils sharply increased after infection. Moreover, dendritic cells, macrophages, and basophils showed inhibitory interactions with exhausted CD4+ T cells. Besides, in mouse livers, we found that exhausted CD4+ T cells were distributed around egg granuloma, promoting collagen expression in primary mouse hepatic stellate cells via IL-4 secretion, resulting in liver fibrosis. Our study provides comprehensive characterization of the composition and cellular states of immune cells with disease progression, which will facilitate better understanding of the mechanism underlying liver fibrotic granulomatous response in schistosomiasis.

LncRNA HOXD-AS1 promotes oral squamous cell carcinoma by sponging miR-203a-5p.

OBJECTIVE: In the present study, we aimed to explore lncRNA HOXD cluster antisense RNA 1 (HOXD-AS1) expression in oral squamous cell carcinoma (OSCC) tissues, its biological roles, and the underlying potential mechanisms in OSCC progression. MATERIALS AND METHODS: HOXD-AS1 expression in paired OSCC and non-tumor tissues from 60 OSCC patients was determined by RT-qPCR. The effects of HOXD-AS1 and miR-203a-5p overexpression on expression of Annexin A4, a validated target of miR-203a-5p, were analyzed by RT-qPCR and Western blot. The roles of HOXD-AS1, miR-203a-5p, and Annexin A4 in the invasion and migration of OSCC cells were analyzed by Transwell assays. RESULTS: HOXD-AS1 overexpression in OSCC predicted poor survival. HOXD-AS1 was predicted to interact with miR-203a-5p, but its expression was not significantly correlated with miR-203a-5p. HOXD-AS1 overexpression increased Annexin A4 expression, while miR-203a-5p overexpression decreased Annexin A4 expression in OSCC cells. Transwell assays showed that invasion and migration of OSCC cells were enhanced by HOXD-AS1 and Annexin A4 overexpression but were reduced by miR-203a-5p overexpression. In addition, miR-203a-5p overexpression suppressed the role of HOXD-AS1 in cell movement and Annexin A4 expression. CONCLUSIONS: HOXD-AS1 may upregulate Annexin A4 by sponging miR-203a-5p in OSCC to promote cancer cell invasion and migration.

The application of single beta-human chorionic gonadotropin (ß-hCG) level measurement in women undergoing single blastocyst transfer.

We previously explored the associations between ß-hCG on the 14th day post-embryo transfer (ET) and reproductive outcomes and established a series of cutoff values to predict different outcomes. The aim of this study was to explore the parameters associated with ß-hCG levels and establish ß-hCG cutoff values in women undergoing single blastocyst transfer. The patients were transferred with either fresh or frozen-thawed blastocysts. Serum ß-hCG levels were compared among different groups. Cutoff values of ß-hCG were established and applied to divide the patients into different groups, among which the ß-hCG groups were compared. Develop day negatively affected ß-HCG levels in those who were pregnant or gave live birth (P < 0.001, 0.008). Inner cell mass significantly affected ß-hCG levels in women who were pregnant or gave live birth (P = 0.013, 0.044). Trophectoderm significantly affected ß-hCG levels in women with most reproductive outcomes, except biochemical pregnancy (BP) (P = 0.184). The cutoff values of ß-hCG for predicting positive outcomes were 194.1, 503.0, 1048.0, and 2590.5 mIU/L. BP rates and adverse pregnancy outcome rates were significantly lower in the higher ß-hCG groups (P < 0.001). Shorter gestational age and lower birth weight and length (P = 0.005, 0.041, 0.003) were observed in the lowest-concentration ß-hCG group. The application of a single ß-hCG measurement was sufficient to predict reproductive outcome in women undergoing blastocyst transfer, under the full consideration of blastocyst parameters. However, the association between ß-hCG and obstetric outcomes remains to be investigated and fully explained.

FOXF1 Was Identified as a Novel Biomarker of Infantile Hemangioma by Weighted Coexpression Network Analysis and Differential Gene Expression Analysis.

Background: The most frequent benign tumor in newborns is infantile hemangioma. The majority of infantile hemangiomas has a favorable prognosis and generally fades away on their own. Some people, however, do experience major consequences. Transcriptome alterations in infantile hemangiomas are yet unclear. The use of transcriptome analysis to uncover diagnostic markers for infantile hemangioma has clinical implications. Methods: The dataset GSE127487 for infantile hemangioma was obtained from the GEO database. The gene set most related with infantile hemangioma was investigated using weighted coexpression network analysis. Differential expression analysis was performed to see whether genes were up or downregulated in infantile hemangiomas. The enrichment of gene sets in pathways or functions is determined via enrichment analysis. Hub genes were discovered via protein-protein interaction network analysis. The relationship between hub genes and immune cells was investigated using immunomicroenvironment analysis. Results: Turquoise and Pink modules were revealed to be the most linked with infantile hemangioma in a weighted coexpression network analysis (p < 0.001). The genes in the two modules were mostly concentrated in actin filament organization, embryonic organ development, reproductive structure development, cell substrate adhesion, extracellular matrix organization, and so on, according to GO enrichment analysis (p < 0.05). These gene enrichment pathways comprised the PI3K-Akt signaling pathway, human papillomavirus infection, focal adhesion, and hepatitis C pathways, according to KEGG enrichment analyzes (p < 0.05). Differential expressed gene analysis showed 43 upregulated and 21 downregulated genes in infantile hemangiomas. We found the gene set most associated to infantile hemangioma by intersecting the elevated genes with the genes acquired by WGCNA, with FOXF1 serving as the hub gene. FOXF1 was linked to the degree of monocyte infiltration, according to immunocorrelation analysis (p < 0.05). B cell memory, dendritic cells resting, macrophage M0, neutrophils, and T cells helper are all negatively connected (p < 0.05). In the FOXF1 hyperexpression group, GSEA analysis revealed that cholesterol homeostasis and cell cycle-associated pathways G2M checkpoint were primarily activated (p < 0.05). Conclusion: FOXF1 was found to be a reliable biomarker of infantile hemangiomas in our research of transcriptome changes in infantile hemangiomas.

Proteomics-based evaluation of the mechanism underlying vascular injury via DNA interstrand crosslinks, glutathione perturbation, mitogen-activated protein kinase, and Wnt and ErbB signaling pathways induced by crotonaldehyde.

Crotonaldehyde (CRA)-one of the major environmental pollutants from tobacco smoke and industrial pollution-is associated with vascular injury (VI). We used proteomics to systematically characterize the presently unclear molecular mechanism of VI and to identify new related targets or signaling pathways after exposure to CRA. Cell survival assays were used to assess DNA damage, whereas oxidative stress was determined using colorimetric assays and by quantitative fluorescence study; additionally, cyclooxygenase-2, mitogen-activated protein kinase pathways, Wnt3a, ß-catenin, phospho-ErbB2, and phospho-ErbB4 were assessed using ELISA. Proteins were quantitated via tandem mass tag-based liquid chromatography-mass spectrometry and bioinformatics analyses, and 34 differentially expressed proteins were confirmed using parallel reaction monitoring, which were defined as new indicators related to the mechanism underlying DNA damage; glutathione perturbation; mitogen-activated protein kinase; and the Wnt and ErbB signaling pathways in VI based on Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and protein-protein interaction network analyses. Parallel reaction monitoring confirmed significant (p < 0.05) upregulation (> 1.5-fold change) of 23 proteins and downregulation (< 0.667-fold change) of 11. The mechanisms of DNA interstrand crosslinks; glutathione perturbation; mitogen-activated protein kinase; cyclooxygenase-2; and the Wnt and ErbB signaling pathways may contribute to VI through their roles in DNA damage, oxidative stress, inflammation, vascular dysfunction, endothelial dysfunction, vascular remodeling, coagulation cascade, and the newly determined signaling pathways. Moreover, the Wnt and ErbB signaling pathways were identified as new disease pathways involved in VI. Taken together, the elucidated underlying mechanisms may help broaden existing understanding of the molecular mechanisms of VI induced by CRA.

Ambient ozone, and urban PM2.5 co-exposure, aggravate allergic asthma via transient receptor potential vanilloid 1-mediated neurogenic inflammation.

Allergic asthma is the most common pulmonary inflammatory disease, and epidemiological studies have revealed that PM2.5 or ambient ozone (O3) exposure contribute to the higher prevalence of allergic asthma. Current experimental evidence focus principally on the pathogenic effect of exposure to a single air pollutant, ignoring the possible synergistic effect of combined exposure to a mix of these pollutants, which is a more realistic scenario. In this study, allergic mice and a nociceptor antagonist were used to explore the mechanisms of co-exposure to these two important air pollutants. Compared with exposure to either PM2.5 or O3, combined exposure to both greatly aggravated allergic asthma in a dose dependent manner, including increased airway hyperresponsiveness, goblet cell metaplasia, more severe airway inflammation and higher oxidative stress levels. In addition, co-exposure in the allergic mice resulted in elevation of the expression of transient receptor potential vanilloid 1 (TRPV1), and of the production of substance P (SP), which exacerbated lung inflammation by neurogenic inflammation. TRPV1 antagonist (capsazepine, CPZ) treatment for the co-exposed allergic mice, markedly attenuated TRPV1 expression and SP release, and reduced airway inflammation and oxidative damage, further alleviating airway hyperresponsiveness. We conclude that neuro-immune interactions might be involved in PM2.5 and O3 co-exposure aggravated allergic asthma.

Ultrasonic Image Feature Analysis under Deep Learning Algorithm to Evaluate the Efficacy of Drug-Coated Balloon for Treatment of Arteriosclerotic Occlusion.

In order to explore the adoption of ultrasonic images under deep learning (DL) algorithm to evaluate the efficacy of drug-coated balloon (DCB) for treatment of arteriosclerotic occlusion, 56 patients who underwent DCB surgery of lower limb artery were selected and all the patients received the examinations of algorithmic ultrasound and digital subtraction angiography (DSA) before surgery. According to the examination methods, they were classified into algorithmic ultrasound group and DSA group. One to two months after DCB surgery, ultrasound examination was performed with the region-based faster convolutional neural network (faster R-CNN) target detection algorithm to check the therapeutic effect. The results showed that the image effect processed by the target detection algorithm based on DL was signally better than that of traditional ultrasonic processing algorithm in Dice, precision (Pre), and sensitivity, with significant difference (P < 0.05). Compared with DSA, algorithmic ultrasound showed better consistency between the two groups in the diagnosis of common femoral artery, superficial femoral artery, and popliteal artery stenosis, with statistical significance (P < 0.05). However, for the diagnosis of anterior tibial artery stenosis, the consistency between algorithmic ultrasound and DSA was general. The residual stenosis of each artery segment decreased obviously in postoperative review compared with that before surgery, and the difference was statistically significant (P < 0.05). Besides, both the pulsatility index (PI) and the blood flow velocity of the dorsalis pedis artery increased after surgery, compared with those before surgery, with significant differences (P < 0.05). To sum up, ultrasound based on DL target detection algorithm had good imaging effect and good consistency with DSA, which was of the clinical reference value. Additionally, DCB surgery was helpful to treat arteriosclerosis occlusion and improve limb blood supply, which had clinical adoption value.

Liver cancer heterogeneity modeled by in situ genome editing of hepatocytes.

Mechanistic study and precision treatment of primary liver cancer (PLC) are hindered by marked heterogeneity, which is challenging to recapitulate in any given liver cancer mouse model. Here, we report the generation of 25 mouse models of PLC by in situ genome editing of hepatocytes recapitulating 25 single or combinations of human cancer driver genes. These mouse tumors represent major histopathological types of human PLCs and could be divided into three human-matched molecular subtypes based on transcriptomic and proteomic profiles. Phenotypical characterization identified subtype- or genotype-specific alterations in immune microenvironment, metabolic reprogramming, cell proliferation, and expression of drug targets. Furthermore, single-cell analysis and expression tracing revealed spatial and temporal dynamics in expression of pyruvate kinase M2 (Pkm2). Tumor-specific knockdown of Pkm2 by multiplexed genome editing reversed the Warburg effect and suppressed tumorigenesis in a genotype-specific manner. Our study provides mouse PLC models with defined genetic drivers and characterized phenotypical heterogeneity suitable for mechanistic investigation and preclinical testing.