Identification of a novel endocytosis­associated gene signature for prognostic prediction in lung adenocarcinoma.

Lung cancer is one of the most common malignant solid tumors and the leading cause of cancer-associated mortality worldwide. Endocytosis is an essential physiological activity for cells to maintain membrane homeostasis, and has been reported to serve an important role in tumorigenesis and progression. In the present study, the aim was to construct a prognostic prediction model of endocytosis-associated genes for patients with lung adenocarcinoma (LUAD). The endocytosis-associated gene signature was established using Lasso Cox regression analysis using the training set of the LUAD cohort from The Cancer Genome Atlas (TCGA) database, and verified using two datasets from the Gene Expression Omnibus (GEO) database. Kaplan-Meier survival curves were used to evaluate the effectiveness of the prognostic evaluation of patients with LUAD. Differentially expressed genes were screened in the tumor tissue of patients compared with paired paracancerous tissues. A series of candidate genes associated to the prognosis of patients with LUAD was obtained using univariate Cox's regression analysis. Using the Lasso Cox regression analysis, an appropriate risk model with 18 endocytosis-associated genes was established. A high-risk score was positively correlated with a higher tumor stage and pathologic grade. Patients with LUAD and high-risk scores had shorter survival times, increased intratumor heterogeneities and immune cell infiltration into tumor tissues, compared with those patients with LUAD and low-risk scores. The endocytosis inhibitor chloroquine could repress proliferation and increase the apoptosis of lung cancer cells. In summary, a novel endocytosis-associated gene signature was constructed using TCGA and GEO datasets. Patients with LUAD and high-risk scores, as calculated by the signature, had a poor prognosis and short survival time.

ResNet14Attention network for identifying the titration end-point of potassium dichromate.

With the rapid development of industry, the increasing discharge of sewage causes the detection of water quality to be of increasing importance. Potassium dichromate titration is one of the most important testing methods in water quality detection; the ability to accurately identify the titration end-point of potassium dichromate is currently a research challenge. To identify titration end-point quickly and accurately, this study proposes a ResNet14Attention network, which utilizes residual modules that focus on original image information and an attention mechanism that focuses highly on classification targets. The proposed ResNet14Attention network is compared with 12 convolutional neural networks such as ResNet series networks, VGG, and GoogLeNet. The results of comparison experiments reveal that only the proposed ResNet14Attention network has the highest training and testing accuracy of 100% among all convolutional neural networks in the comparison experiment; the proposed ResNet14Attention network has the highest training speed compared to all the networks that over 90% accuracy.

Quantification of Biologically Active DNA Alkylation in Temozolomide-Exposed Glioblastoma Cell Lines by Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry: Method Development and Recommendations for Validation.

Quantitative monitoring of biologically active methylations of guanines in samples exposed to temozolomide (TMZ) would be useful in glioblastoma research for preclinical TMZ experiments, for clinical pharmacology questions regarding appropriate exposure, and ultimately for precision oncology. The known biologically active alkylation of DNA induced by TMZ takes place on O6 position of guanines. However, when developing mass spectrometric (MS) assays, the possible signal overlap of O6-methyl-2'-deoxyguanosine (O6-m2dGO) with other methylated 2'-deoxyguanosine species in DNA and methylated guanosines in RNA must be considered. Liquid chromatography-tandem MS (LC-MS/MS) offers the analytical requirements for such assays in terms of specificity and sensitivity, especially when multiple reaction monitoring (MRM) is available. In preclinical research, cancer cell lines are still the gold standard model for in vitro drug screening. Here, we present the development of ultra-performance LC-MRM-MS assays for the quantification of O6-m2dGO in a TMZ-treated glioblastoma cell line. Furthermore, we propose adapted parameters for method validation relevant to the quantification of drug-induced DNA modifications.

Desorption Kinetics Evaluation for the Development of Validated Desorption Electrospray Ionization-Mass Spectrometric Assays for Drug Quantification in Tissue Sections.

The development of desorption/ionization (DI) mass spectrometric (MS) assays for drug quantification in tissue sections and their validation according to regulatory guidelines would enable their universalization for applications in (clinical) pharmacology. Recently, new enhancements in desorption electrospray ionization (DESI) have highlighted the reliability of this ion source for the development of targeted quantification methods that meet requirements for method validation. However, it is necessary to consider subtle parameters leading to the success of such method developments, such as the morphology of desorption spots, the analytical time, and sample surface, to cite but a few. Here, we provide additional experimental data highlighting an additional important parameter, based on the unique advantage of DESI-MS on continuous extraction during analysis. We demonstrate that considering desorption kinetics during DESI analyses would largely help (i) reducing analytical time during profiling analyses, (ii) verifying solvent-based drug extraction using the selected sample preparation method for profiling and imaging modes, and (iii) predicting the feasibility of imaging assays using samples in a given expected concentration range of the targeted drug. These observations will likely serve as precious guidance for the development of validated DESI-profiling and imaging methods in the future.

Five-years' prognostic analysis for coronary artery ectasia patients with coronary atherosclerosis: A retrospective cohort study.

Background: Most of coronary artery ectasia (CAE) patients have comorbid coronary atherosclerosis. It was lack of prognostic data for CAE patients with coronary heart disease (CHD) and for whom with acute myocardial infarction (AMI). Objective: To determine the overall prognosis for CAE patients. Materials and methods: This study was a retrospective cohort study. Fifty-one patients with CAE and comorbid AMI (CAE + AMI) and 108 patients with CAE and comorbid CHD (CAE + CHD) were enrolled and matched to non-CAE subjects at a ratio of 1:3 using a propensity score method, respectively. Controls for CAE + AMI group were 153 AMI patients, controls for CAE group were 324 CHD patients and 329 participants with relatively normal coronary arteries (CON). We followed them up to observe major cardiovascular events (MACE). Results: The Kaplan-Meier curves showed that the prognosis in CAE + AMI group was worse than in AMI group (5-year non-MACE rate: 62.70% vs. 79.70%, P = 0.010), the prognosis in CAE group was worse than in CHD and CON groups (5-year non-MACE rate: 74.10% vs. 85.80% and 96.70%, respectively, P = 0.000). The main MACEs in CAE + AMI and CAE groups were AMI reoccurrence (19.61% vs. 4.57%, P = 0.002) and re-hospitalization due to repeated angina pectoris (14.81% vs. 8.33% and 2.74%, P = 0.000), respectively. Additionally, the COX regression analysis revealed that the protective factors for preventing MACE in CAE + AMI group included antiplatelet agents (hazard ratio = 0.234, P = 0.016) and angiotensin-converting enzyme inhibitor/angiotensin receptor inhibitor (ACEI/ARB, hazard ratio = 0.317, P = 0.037). Whereas the main factor promoting MACE in CAE group was the degree of coronary stenosis (Gensini score, hazard ratio = 1.011, P = 0.022). Conclusion: The prognosis of patients with CAE + AMI was worse than that of those with AMI. The overall prognosis of patients with CAE was worse than that of those with CHD. CAE + AMI and CAE groups had different characteristics; the former was prone to AMI reoccurrence, and the latter was prone to repeated angina pectoris. To prevent MACE, medications, including antiplatelets and ACEI/ARBs, are indicated for patients with CAE + AMI, whereas prevention of the progression of atherosclerotic lesions is indicated for patients with CAE.

Neutrophil-to-lymphocyte ratio predicts coronary artery lesion severity and long-term cardiovascular mortality in patients with unstable angina pectoris.

OBJECTIVES: Neutrophil-to-lymphocyte ratio (NLR), one of the composite biomarker of systemic inflammatory status, was proved promising in predicting clinical outcomes of acute coronary syndrome (ACS). However, there were no evidences that NLR was directly relative to the clinical outcomes of unstable angina pectoris (UAP). Therefore, this study was aimed to detect whether NLR could predict the coronary artery lesion severity (indicated as SYNTAX score) and clinical outcomes (especially long-term cardiovascular mortality) in patients with. METHODS: In the single-centre retrospective study, 4110 patients with UAP were enrolled and divided into two groups according to their primary NLR values and followed up at a median time duration of 36 months. The differences of SYNTAX score and cardiovascular mortality between groups were analysed, and the predictive value of NLR was determined. RESULTS: NLR was positively and linearly correlated with SYNTAX score (r = 0.270). Diabetes (p = 0.049), lymphocyte (p = 0.004), NLR (p = 0.002) and SYNTAX score (p < 0.001) were independent predictors of long-term cardiovascular mortality in patients with UAP. Kaplan-Meier analysis revealed higher occurrence of cardiovascular mortality when NLR > 2.38 (p = 0.015). Receiver operating characteristic (ROC) analysis showed that NLR = 2.76 is an effective cut point for predicting cardiovascular mortality (69.2% sensitivity, 64.8% specificity). CONCLUSIONS: NLR value was positively related to the severity of coronary artery lesion and proved to be an independent predictor of cardiovascular mortality in patients with UAP. This study would contribute to therapy and prognosis optimisation of UAP.

Analytical Performance Evaluation of New DESI Enhancements for Targeted Drug Quantification in Tissue Sections.

Desorption/ionization (DI)-mass spectrometric (MS) methods offer considerable advantages of rapidity and low-sample input for the analysis of solid biological matrices such as tissue sections. The concept of desorption electrospray ionization (DESI) offers the possibility to ionize compounds from solid surfaces at atmospheric pressure, without the addition of organic compounds to initiate desorption. However, severe drawbacks from former DESI hardware stability made the development of assays for drug quantification difficult. In the present study, the potential of new prototype source setups (High Performance DESI Sprayer and Heated Transfer Line) for the development of drug quantification assays in tissue sections was evaluated. It was demonstrated that following dedicated optimization, new DESI XS enhancements present promising options regarding targeted quantitative analyses. As a model compound for these developments, ulixertinib, an inhibitor of extracellular signal-regulated kinase (ERK) 1 and 2 was used.

Antithrombotic treatment strategy for patients with coronary artery ectasia and acute myocardial infarction: A case report.

BACKGROUND: There is no consensus on the antithrombotic treatment strategy for patients with coronary artery ectasia (CAE). CASE SUMMARY: This case reports the dynamic observation of a patient for 48 mo after a diagnosis of CAE with acute myocardial infarction (AMI). The first antithrombotic agents used were aspirin (100 mg/d) and clopidogrel (75 mg/d). During the sixth month of observation, a second AMI occurred involving the same culprit vessel; therefore, antithrombotic agents were changed to aspirin (100 mg/d) and ticagrelor (90 mg twice per day). Twelve months after the second AMI, an attempt to reduce the dosage ticagrelor failed; therefore the original dose was continued. The CAE was relatively stable during the following 4 years. CONCLUSION: This case indicates that a combination of aspirin and ticagrelor may be more effective for CAE patients with AMI than aspirin and clopidogrel.

The uric acid to albumin ratio: a novel predictor of long-term cardiac mortality in patients with unstable angina pectoris after percutaneous coronary intervention.

The prognosis of unstable angina pectoris (UAP) differs from non-ST-segment elevation myocardial infarction, and percutaneous coronary intervention (PCI) is considered to improve outcomes of UAP. This study aimed to assess the prognostic value of uric acid to albumin ratio (UAR) for long-term mortality in UAP patients after PCI. Our study retrospectively enrolled 2298 patients hospitalized because of UAP in a tertiary hospital. Divided by medium UAR, the patients were classified into two groups. Baseline demographics, clinical features and laboratory characteristics were obtained from medical records. Post-discharge follow-up was performed either in outdoor clinic or through phone call. The primary endpoint in this study was cardiac death, while all-cause death and rehospitalization were designated as the secondary endpoints. The median follow-up time was 672 days. Among all patients, 58 (2.5%) died, 28 of which died of cardiac deaths (1.2%), and 467 were re-hospitalized (20.3%). Cardiac mortality and all-cause mortality were found to be significantly higher in the high UAR group than in the low UAR group (p = 0.007, p < 0.001), and Kaplan-Meier analysis showed patients with higher UAR may suffer from worse outcomes (p = 0.020). UAR, PCI history, and age were identified as independent predictors of cardiac mortality by multivariate Cox regression. A UAR value of >8.35 was demonstrated as an ideal cut-off point to predict post-PCI cardiac mortality (p <0.001). Overall, it is indicated that baseline UAR was independently correlated with long-term cardiac mortality in patients with UAP treated by PCI.

Is Coronary Artery Ectasia a Progressive Disease? A Self-Controlled Retrospective Cohort Study.

Objective: It is essential to understand whether coronary artery ectasia (CAE) progresses over time because the patients might be under the risk of coronary rupture, and stent implant should be avoided if ectatic changes progress. Methods: A consecutive series of 99 CAE patients who had undergone coronary angiography at least twice were enrolled and followed up for 1-16 years until they received a second angiogram. Subjects were divided into two groups (1-5 vs. 5-16 years of follow-up), then the basic clinical characteristics and coronary artery images were compared over time. Results: (1) All CAE patients exhibited atherosclerosis, and a majority presented with acute myocardial infarction. Most baseline clinical characteristics were relatively stable. (2) Atherosclerosis (indicated by the distribution of stenosis in coronary vessels) and the Gensini scores progressed significantly. Ectasia extent showed minimal changes as indicated by blood vessel involvement, Markis type, coronary blood flow, ectasia diameter, and ectasia length. (3) Multilinear regression analysis revealed that the underlying factors related to stenosis evolution indicated by fold of Gensini score were: longer time interval, lower baseline Gensini score, and higher hypersensitive C-reactive protein concentration. (4) There was a relationship between the ectatic diameter and the extent of stenosis. Conclusions: For CAE patients with atherosclerosis followed for 1-16 years, there was minimal CAE progression, while the atherosclerosis progressed and the ectasia extent was related to degree of stenosis. The results indicate that prevention and treatment of atherosclerotic changes might have more clinical significance than addressing ectatic changes.

Urinary Alpha1-Microglobulin: A New Predictor for In-Hospital Mortality in Patients with ST-Segment Elevation Myocardial Infarction.

BACKGROUND Alpha1-microglobulin (A1MG) is a small molecular protein related to oxidation and inflammation. It exists in diverse body fluids, including urine. Results from urine tests are sometimes neglected when predicting in-hospital prognosis. It remains unclear whether urinary A1MG (UA1MG) can predict short-term prognosis of ST-elevated myocardial infarction (STEMI). MATERIAL AND METHODS A total of 1854 hospitalized patients with acute STEMI were retrospectively enrolled in our study. Medical records were used to obtain patient demographic and clinical information, UA1MG values (which were used to divide patients into groups of low, medium, or high), and other laboratory parameters. Principal clinical outcomes of interest were all-cause in-hospital deaths, cardiac deaths, and major adverse cardiac events (MACEs). RESULTS Among the 1854 enrolled patients, 43 (2.3%) died in the hospital, of which 33 (1.8%) were cardiac deaths. MACEs were noted in 113 patients (6.1%) during hospitalization. The group with the highest UA1MG value showed a significantly higher frequency of in-hospital deaths, cardiac deaths, and MACEs, compared to those of the lowest UA1MG value group (4.4% vs. 1.0%, P<0.001; 3.1% vs. 0.6%, P<0.005; and 8.6% vs. 4.7%, P=0.007, respectively). Multivariate regression analysis revealed that UA1MG levels (odds ratio 1.109, 95% confidence interval (CI) 1.027-1.197, P=0.008) independently predicted all-cause in-hospital mortality. A UA1MG value of 3.23 mg/dL was considered as an optimal cutoff point in STEMI to predict all-cause mortality after receiver operating characteristic curve analysis (area under the curve 0.73, 95% CI 0.65-0.80, P<0.001). CONCLUSIONS The UA1MG value at hospital admission could be an independent prognostic factor of all-cause in-hospital mortality in patients with STEMI.

Peripheral blood soluble elastin and elastase as auxiliary diagnostic indicators for coronary artery ectasia

Background/aim: Damage to elastin fibres in coronary media might lead to coronary artery ectasia (CAE). This study evaluated whether CAE can be distinguished by detecting circulating soluble elastin (s-elastin), which is a degradation product of elastin fibres, and elastase, which is the main enzyme of elastin fibres. Materials and methods: Fifty-eight patients with CAE, 58 with coronary heart disease (CHD) and 61 with relatively normal coronary arteries, were included. Circulating s-elastin and elastase were measured, and receiver operating characteristic curves were used to demonstrate their respective optimal cut-off values for predicting CAE. Results: The concentrations of s-elastin and elastase were higher in the CAE group than in the CHD and relatively-normal-coronary groups. Their cut-off values for screening of CAE were 13.148 ng/mL and 25.549 ng/mL, respectively; for sensitivity of CAE were 0.690 and 0.773, respectively; and for specificity of CAE were 0.862 and 0.571, respectively. A combination of s-elastin and elastase in series (one of the two higher than its cut-off value) had a better sensitivity for screening for CAE, whereas their combination in parallel (both higher than their cut-off values) had a better specificity. Conclusion: Circulating s-elastin and elastase are promising biomarkers for assisting in CAE diagnosis.

Increased serum adiponectin predicts improved coronary flow and clinical outcomes in patients with ST-segment elevation myocardial infarction treated by primary percutaneous coronary intervention.

BACKGROUND: Previous studies suggested that adiponectin (APN) could ameliorate ischemia/reperfusion injury and endothelial dysfunction in patients with acute myocardial infarction. However, the relationship between serum APN level and coronary flow after primary percutaneous coronary intervention (PPCI) in patients with ST-segment elevation myocardial infarction (STEMI) is unclear. METHODS: A total of 144 patients with STEMI treated by PPCI were enrolled and divided into two groups based on the mean serum APN level on admission. The data on coronary angiograms and laboratory examinations were collected and compared between groups. The incidence of major adverse cardiac events (MACE) was evaluated in all enrolled patients. RESULTS: The prevalence of Thrombolysis In Myocardial Infarction (TIMI) flow grade <3 after PPCI and corrected TIMI frame count were lower in the high-APN group (P = 0.032 and P = 0.029, respectively). Logistic regression analysis demonstrated that APN was an independent negative predictor of poor coronary flow after PPCI (odds ratio = 0.72, 95% CI: 0.56-0.93, P = 0.011). Kaplan-Meier curves showed that a higher APN level correlated with a better MACE-free survival rate, and multivariate Cox hazard regression analysis indicated that high APN was a significant negative predictor of MACE (hazard ratio = 0.54, 95% CI: 0.29-1.00, P = 0.048). CONCLUSION: Elevated serum levels of APN on admission are associated with improved myocardial blood flow and clinical outcomes in STEMI patients treated with PPCI.

Exosomes derived from SDF1-overexpressing mesenchymal stem cells inhibit ischemic myocardial cell apoptosis and promote cardiac endothelial microvascular regeneration in mice with myocardial infarction.

Exosomes extracted from mesenchymal stem cells (MSCs) was reported to reduce myocardial ischemia/reperfusion damage. Besides, stromal-derived factor 1 (SDF1a) functions as cardiac repair after myocardial infarction (MI). Therefore, the present study aims to identify whether exosomes (Exo) released from SDF1-overexpressing MSCs display a beneficial effect on ischemic myocardial infarction. Initially, a gain-of-function study was performed to investigate the function of SDF1 in ischemic myocardial cells and cardiac endothelial cells. Coculture experiments were performed to measure potential exosomic transfer of SDF1 from MSCs to ischemic myocardial cells and cardiac endothelial cells. During the coculture experiments, exosome secretion was disrupted by neutral sphingomyelinase inhibitor GW4869 and upregulated exosomal SDF1 using SDF1 plasmid. Effects of Exo-SDF1 on cardiac function in MI mice were investigated in vivo. MSCs suppressed myocardial cell apoptosis and promoted microvascular regeneration of endothelial cells through secretion of exosomes. The addition of GW4869 led to increased apoptotic capacity of myocardial cells, decreased microvascular formation ability of endothelial cells, enhanced autophagy ability, and elevated Beclin-1 level as well as ratio of LC3II/LC3I. Overexpression of SDF1 and Exo-SDF1 inhibited apoptosis and autophagy of myocardial cells, but promoted tube formation of endothelial cells. The interference of PI3K signaling pathway promoted apoptosis and autophagy of myocardial cells, but inhibited tube formation of endothelial cells. SDF1 activated the PI3K signaling pathway. Exo-SDF1 protected cardiac function of MI mice and inhibited myocardial tissue damage. This study provided evidence that SDF1 overexpression in MSCs-derived exosomes inhibited autophagy of ischemic myocardial cells and promoted microvascular production of endothelial cells.

Is Coronary Artery Ectasia a Thrombotic Disease?

Coronary artery ectasia (CAE) is a rare disease and a substantial portion of patients with CAE are first diagnosed with acute myocardial infarction (AMI). The question was raised if CAE was a kind of thrombotic disease. We assessed a consecutive series of 119 patients with CAE including 32 patients with AMI (CAE + AMI group) and 87 patients without AMI (CAE group). During the same period, 90 patients with coronary heart disease, 90 patients with normal coronary arteries (control), and 120 AMI patients without CAE (AMI group) were randomly selected and evaluated. Both current and previous AMI prevalence rates in the CAE population were higher than the AMI rate for the other patients undergoing coronary angiograms; the mean platelet volume and fibrinogen were increased in the CAE + AMI and CAE groups. For patients with CAE with AMI, most of the thrombotic lesions were in the ectasia site. After dividing the patients with CAE into with and without antiplatelet treatment groups before admission, the AMI rate was lower in the antiplatelet group. Platelets may participate in the thrombotic process in CAE. Antiplatelet treatment may decrease the AMI rate of patients with CAE.

Impact of red blood cell distribution width on long-term mortality in patients with ST-elevation myocardial infarction.

OBJECTIVES: Recent studies have reported increased red blood cell distribution width (RDW) has been associated with adverse outcomes in heart failure and stable coronary disease. We investigated the association between RDW and risk of all-cause mortality in patients with ST-elevation myocardial infarction (STEMI) who were free of heart failure at baseline. METHODS: We enrolled 691 patients with STEMI who were free of heart failure at baseline confirmed by coronary angiography in Beijing Friendship Hospital from January 2007 to December 2008. According to the median RDW at baseline (13.0%) on admission, the patients were divided into two groups: a low-RDW group (RDW <13.0%, n = 329) and a high-RDW group (RDW ≥13.0%, n = 362). All-cause mortality rates were compared between groups. Mean duration of follow-up was 41.8 months. The relation between RDW and clinical outcomes after hospital discharge were tested using Cox regression models, adjusting for clinical variables. At the same time, the sensitivity and specificity of RDW were analyzed by ROC analysis. RESULTS: Forty-seven patients (6.8%) died during follow-up. The cumulative incidence of all-cause death was significantly higher in the high-RDW group than in the low-RDW group (log-rank p = 0.007). Multivariate analysis revealed that high RDW was associated with all-cause mortality (hazard ratio: 3.43; 95% confidence interval: 1.17-8.32; p = 0.025). The area under the ROC curve was 0.562. CONCLUSION: From the statistical point of view, increased RDW is associated with all-cause and cardiac mortality rates in patients with STEMI who were free of heart failure at baseline. But RDW is a marker with a very low prognostic accuracy that does not seem to be clinically helpful.