Proteomic analysis of plasma proteins from patients with cardiac rupture after acute myocardial infarction using TMT-based quantitative proteomics approach.

BACKGROUND: Cardiac rupture (CR) is a rare but catastrophic mechanical complication of acute myocardial infarction (AMI) that seriously threatens human health. However, the reliable biomarkers for clinical diagnosis and the underlying signaling pathways insights of CR has yet to be elucidated. METHODS: In the present study, a quantitative approach with tandem mass tag (TMT) labeling and liquid chromatography-tandem mass spectrometry was used to characterize the differential protein expression profiles of patients with CR. Plasma samples were collected from patients with CR (n = 37), patients with AMI (n = 47), and healthy controls (n = 47). Candidate proteins were selected for validation by multiple reaction monitoring (MRM) and enzyme-linked immunosorbent assay (ELISA). RESULTS: In total, 1208 proteins were quantified and 958 differentially expressed proteins (DEPs) were identified. The difference in the expression levels of the DEPs was more noticeable between the CR and Con groups than between the AMI and Con groups. Bioinformatics analysis showed most of the DEPs to be involved in numerous crucial biological processes and signaling pathways, such as RNA transport, ribosome, proteasome, and protein processing in the endoplasmic reticulum, as well as necroptosis and leukocyte transendothelial migration, which might play essential roles in the complex pathological processes associated with CR. MRM analysis confirmed the accuracy of the proteomic analysis results. Four proteins i.e., C-reactive protein (CRP), heat shock protein beta-1 (HSPB1), vinculin (VINC) and growth/differentiation factor 15 (GDF15), were further validated via ELISA. By receiver operating characteristic (ROC) analysis, combinations of these four proteins distinguished CR patients from AMI patients with a high area under the curve (AUC) value (0.895, 95% CI, 0.802-0.988, p < 0.001). CONCLUSIONS: Our study highlights the value of comprehensive proteomic characterization for identifying plasma proteome changes in patients with CR. This pilot study could serve as a valid foundation and initiation point for elucidation of the mechanisms of CR, which might aid in identifying effective diagnostic biomarkers in the future.

Potent induction of antitumor immunity by combining cryo-thermal ablation with immune checkpoint inhibitors in hepatocellular carcinoma.

BACKGROUND: The low response rate of immune checkpoint inhibitors (ICIs) prompts the exploration of novel combination therapies for patients with hepatocellular carcinoma (HCC). Here, we aimed to examine the efficiency and potential mechanism of cryo-thermal ablation (Cryo-A) combined with anti-programmed death protein 1 (αPD1) and/or cytotoxic T-lymphocyte antigen 4 (αCTLA4) inhibitors in a murine hepatoma model. METHOD: Immunocompetent C57BL/6 mice inoculated with unilateral or bilateral H22 hepatic tumour cells were treated with Cryo-A and/or ICIs (αPD1 and/or αCTLA4). Flow cytometry, immunohistochemistry, ELISpot assay, time-of-flight cytometry, tumour rechallenging, and T-cell depletion assay were used to assess the dynamic changes of immune cell subsets following therapy. RESULTS: We found Cryo-A resulted in immunogenic cell death of tumour cells, activation of dendritic cells, and enhancement of antitumor immunity. Cryo-A alone was insufficient to extend survival, combining Cryo-A with αPD1 and αCTLA4 further modulated the tumour microenvironment, inducing a durable antitumor immune response by tumour-reactive CD8+ T cells and significantly prolonged survival. Time-of-flight cytometry (CyTOF) data revealed that combination therapies reshaped the tumour microenvironment by the increase of intratumoral CD8+ T cells expressed higher levels of cytotoxic markers and immune checkpoint molecules, and by downregulation of intratumoral granulocytes. The combination also resulted in the eradication of remote unablated tumours (abscopal effect). CONCLUSIONS: These findings suggested that Cryo-A turned HCC from "cold" tumours to "hot" tumours and the combination of Cryo-A with αPD1 and αCTLA4 may be a promising approach to improve the prognosis of HCC.

Bufalin Inhibits Tumorigenesis and SREBP-1-Mediated Lipogenesis in Hepatocellular Carcinoma via Modulating the ATP1A1/CA2 Axis.

Altered lipid metabolism is a hallmark of hepatocellular carcinoma (HCC), a common malignancy with a dismal prognosis against which there is a lack of effective therapeutic strategies. Bufalin, a classical Na[Formula: see text]-K[Formula: see text]-ATPase (NKA) inhibitor, shows a potent antitumor effect against HCC. However, the role of bufalin in regulating lipid metabolism-related pathways of HCC remains unclear. In this study, we examined the interaction between bufalin and its target molecule, ATP1A1/CA2, in vitro and in vivo and explored the intersected downstream pathways in silico. A multi-omics analysis of transcriptomics and metabolomics was employed to screen for potential action targets. The results were verified and correlated with the downstream lipid de novo synthesis pathway and the bufalin/ATP1A1/CA2 axis. We found that bufalin suppressed the ATP1A1/CA2 ratio in the treated HCC cells and showed a negative correlation with bufalin drug sensitivity. Functionally, ATP1A1 overexpression and CA2 down-regulation inhibited the bufalin-suppressed HCC proliferation and metastasis. Furthermore, down-regulation of CA2 induced epithelial-mesenchymal transition and bufalin resistance in HCC cells by up-regulating ATP1A1. Mechanistically, lipid metabolism-related signaling pathways were enriched in low ATP1A1 and high CA2 expression subgroups in GSEA. The multi-omics analysis also showed that bufalin was closely related to lipid metabolism. We demonstrated that bufalin inhibits lipogenesis and tumorigenesis by down-regulating SREBP-1/FASN/ACLY via modulating the ATP1A1/CA2 axis in HCC.

Circulating tumor-associated autoantibodies as novel diagnostic biomarkers in pancreatic adenocarcinoma.

To develop a superior diagnostic approach for pancreatic adenocarcinoma (PAAC), the present study prospectively included 338 PAAC patients, 294 normal healthy volunteers (NHV), 122 chronic pancreatitis (CP) patients and 100 patients with non-PAAC malignancies. In the identification phase, HuProt Human Proteome Microarray, comprising 21 065 proteins, was used to identify serum tumor-associated autoantibodies (TAAbs) candidates differentiating PAAC (n = 30) from NHV (n = 30). A PAAC-focused array containing 165 differentially expressed TAAbs identified was subsequently adopted in the validation phase (n = 712) for specificity and sensitivities. The multivariate TAAbs signature for differentiation PAAC from controls (NHV + CP) identified five candidates, namely the IgG-type TAAbs against CLDN17, KCNN3, SLAMF7, SLC22A11 and OR51F2. Multivariate logistic performance model of y = (22.893 × CA19-9 + 0.68 × CLDN17 - 4.012) showed a significant better diagnostic accuracy than that of CA19-9 and CLDN17 in differentiating PAAC from controls (NHV + CP) (AUC = 0.97, 0.92 and 0.82, respectively, P-value < .0001). We further tested the autoantigen level of CLDN17 by ELISA in 82 sera samples from PAAC (n = 42), CP (n = 24) and NHV (n = 16). Similarly, the model showed superior diagnostic performance than that of CA19-9 and CLDN17 (AUC = 0.93, 0.83 and 0.81, respectively, P-value < .0001) in differentiating PAAC from controls. In conclusion, our study is the first to characterize the circulating TAAbs signatures in PAAC. The results showed that CLDN17 combined with CA19-9 provided potentially clinical value and may serve as noninvasive novel biomarkers for PAAC diagnosis.

Identification and Validation of a Tumor Microenvironment-Related Gene Signature in Hepatocellular Carcinoma Prognosis.

Background: Hepatocellular carcinoma (HCC) is a typical inflammatory-related malignant tumor with complex immune tolerance microenvironment and poor prognosis. In this study, we aimed to construct a novel immune-related gene signature for the prognosis of HCC patients, exploring tumor microenvironment (TME) cell infiltration characterization and potential mechanisms. Methods: A total of 364 HCC samples with follow-up information in the TCGA-LIHC dataset were analyzed for the training of the prognostic signature. The Least Absolute Shrinkage and Selector Operation (LASSO) regression based on the IRGs was conducted to identify the prognostic genes and establish an immune risk signature. The immune cell infiltration in TME was estimated via the CIBERSORT method. Gene Set Variation Analysis (GSVA) was conducted to compare the biological pathways involved in the low-risk and high-risk groups. Furthermore, paraffin sections of HCC tissue microarrays containing 77 patients from Fudan University Shanghai Cancer Center were used for IHC staining. The clinical characteristics of the 77 HCC patients were collected and summarized for survival analysis validation via the Kaplan-Meier (KM) method. Results: Three-gene signature with close immune correlation (Risk score = EPO * 0.02838 + BIRC5 * 0.02477 + SPP1 * 0.0002044) was constructed eventually and proven to be an effective prognostic factor for HCC patients. The patients were divided into a high-risk and a low-risk group according to the optimal cutoff, and the survival analysis revealed that HCC samples with high-risk immuno-score had significantly poorer outcomes than the low-risk group (p < 0.0001). The results of CIBERSORT suggested that the immune cell activation was relatively higher in the low-risk group with better prognosis. Besides, GSVA analysis showed multiple signaling differences between the high- and low-risk group, indicating that the three-gene prognostic model can affect the prognosis of patients by affecting immune-related mechanisms. Tissue microarray (TMA) results further confirmed that the expression of three genes in HCC tissues was closely related to the prognosis of patients, respectively. Conclusion: In this study, we constructed and validated a robust three-gene signature with close immune correlation in HCC, which presented a reliable performance in the prediction of HCC patients' survival.

Validity of the Polar V800 Monitor for Assessing Heart Rate Variability in Elderly Adults under Mental Stress and Dual Task Conditions.

Background: Aging may result in autonomic nervous dysfunction. Heart rate variability (HRV) is a non-invasive method to measure autonomic nervous activities. Many studies have shown that HRV contributes to the risk assessment of diseases. A Polar V800 heart rate monitor is a wearable device that measures R-R intervals, but has only been validated in younger adults under limited testing conditions. There is no validation of the V800 under mental stress or in dual task testing conditions. Therefore, this study investigated the validity of the Polar V800 heart rate monitor for assessing R-R intervals and evaluated if there were differences on HRV parameters under different situations in community-dwelling elderly adults. Methods: Forty community-dwelling elderly adults were recruited. Heart rates were recorded via electrocardiogram (ECG) and the V800 under sitting, during an arithmetic test, during a naming test, a self-selected walking velocity test (SSWV), and dual tasks (SSWV performing mental arithmetic test and SSWV performing naming test). Indices of time and frequency domains of HRV were calculated afterwards. The intra-class correlation coefficient (ICC) analysis and effect size were calculated to examine the concurrent validity between the V800 and the ECG. Results: All HRV indices from the V800 were highly correlated with the ECG under all tested conditions (ICC = 0.995-1.000, p < 0.001) and the effect size of bias was small (<0.1). Conclusion: Overall, the V800 has good validity on the assessment of HRV in community-dwelling elderly adults during sitting, mental arithmetic test, naming test, SSWV, and dual tasks.

Retrograde transcatheter closure of ventricular septal perforation after acute myocardial infarction: a case report.

The use of the Lunderquist exchange guide wire via the retrograde approach of the right femoral vein-inferior vena cava-right atrium-right ventricle-ventricular septal perforation-left ventricle-descending aorta can maintain guide wire tension and significantly reduce the operative time. The patient was admitted due to chest pain for 3 hours. The diagnosis was acute anterior septal myocardial infarction with ventricular septal perforation. One week after admission, a drug-eluting stent was implanted in the left anterior descending branch. Repeated echocardiography revealed that the diameter of the ventricular septal perforation had increased from 6 to 12 mm. During this period, the patient suffered from repeated episodes of shortness of breath that were progressively exacerbated. The patient was transferred to the intensive care unit (ICU) and underwent intra-aortic balloon pump (IABP) implantation. Twenty days after admission, the Lunderquist exchange guide wire was used via the retrograde approach of the right femoral vein-inferior vena cava-right atrium-right ventricle-ventricular septal perforation-left ventricle-descending aorta. A 26-mm occluder was released for transcatheter closure of the ventricular septal perforation. Shortness of breath was immediately relieved. The patient was discharged 3 days later. Retrograde transcatheter closure of ventricular septal perforation can effectively reduce operative time and is conducive to quick and stable improvement of the patient's condition.

A Seven-Gene Signature with Close Immune Correlation Was Identified for Survival Prediction of Lung Adenocarcinoma.

BACKGROUND Lung adenocarcinoma is the most life-threatening malignancy with high incidence and poor long-term survival. The crucial role of tumor immunity reveals the significance of exploring immune-related prognostic predictors in lung adenocarcinoma. MATERIAL AND METHODS Immune-related genes were screened out applying the ESTIMATE algorithm. The Cancer Genome Atlas Lung Adenocarcinoma (TCGA-LUAD) dataset was trained for the construction of Cox proportional hazard model. Univariate Cox and Lasso regression analysis was conducted to reduce the overfitting of model. Nomogram integrated the signature and clinicopathological characteristics was established for prognosis prediction of LUAD. The GSE30219, GSE41271, and GSE42127 datasets were analyzed for external validation. LUAD patients were separated into low-risk and high-risk subgroups based on the optimum cutoff threshold of calculated risk score. The predictive value of the signature was evaluated using Kaplan-Meier survival analysis, Harrell's C-index, and receiver operating characteristic (ROC) curve analysis and calibration curve. Clinical- and immune-correlation of the signature was further performed. Gene Set Enrichment Analysis (GSEA) was performed for functional exploration. RESULTS An immune-related signature containing 7 genes was identified. The signature exhibited reliable performance in the prediction of overall survival for LUAD with the C-index being 0.72. The areas under the curve (AUCs) of the model in 1-year risk prediction were 0.781, 0.797, 0.659, and 0.822 for TCGA-LUAD, GSE30219, GSE41271, and GSE42127 datasets, respectively. In all datasets, the signature proved to an independent risk factor for LUAD. Correlation analyses and GSEA further revealed the close relationship between the predictive biomarker and tumor immunity. CONCLUSIONS A Cox proportional hazard model consisting of 7 genes was identified for prognostic prediction of LUAD. The signature was highly correlated with immunity and deserves further exploration.

Cardiac MRI-guided interventional occlusion of ventricular septal rupture in a patient with cobalt alloy stent.

The case of a 68-year-old man with chest pain for 3 days is presented. Coronary angiography demonstrated subtotal occlusion of the mid-left anterior descending artery. A drug-eluting cobalt alloy stent was implanted after balloon dilation. On the 3rd postoperative day, echocardiography showed a ventricular septal rupture (VSR) (7 mm diameter) near the cardiac apex and ventricular aneurysm. On cardiac magnetic resonance imaging (MRI), the VSR was shown to be 11 mm in diameter. The membranous septum was 32 and 27.8 mm along the anteroposterior and superoinferior axes, respectively. The left-to-right shunt was apparent. Four weeks later, interventional therapy was performed to occlude the VSR according to the result of the MRI. The symptoms improved rapidly, and the patient was discharged. At the 4-month follow up visit, cardiac MRI revealed no shunt at the occlusion site, and the edge of the occluder was secured in the adjacent normal cardiac tissues. In conclusion, cardiac MRI could be considered for patients with a newly implanted cobalt alloy stent to provide an accurate assessment of VSR.

The Us2 Gene Product of Herpes Simplex Virus 2 modulates NF-κB activation by targeting TAK1.

HSV-2 is one of the most common sexually transmitted pathogens worldwide and HSV-2 infection triggers cytokine and chemokine production. However, little is known about which HSV-2 genes engage in the regulation of NF-κB signaling and what mechanisms are involved. In a screen of the unique short (Us) regions of HSV-2, we observed that HSV-2 Us2 activates NF-κB signaling. We additionally indicated that deficiencies of Us2 decrease HSV-2 WT mediated NF-κB activation and cytokine and chemokine production, and overexpression of Us2 showed opposite effects. Co-immunoprecipitations indicated that Us2 interacted with TGF-ß activated kinase 1 (TAK1), a serine/threonine kinase essential for NF-κB activation, and Us2 has the ability to regulate the TAK1-mediated pathway and induces TAK1 downstream signaling. Further studies verified that Us2 induced the phosphorylation of TAK1, resulting in the activation of TAK1 mediated downstream signaling. The role of Us2 in HSV-2 induced NF-κB pathways was also confirmed in the Us2-deficient mutant and HSV-2 WT infected mice. Our results indicate that HSV-2 Us2 gene product binds to TAK1 to positively regulate NF-κB signaling and, for the first time, provide insights into the molecular mechanism.