Quercetin induces ferroptosis by inactivating mTOR/S6KP70 pathway in oral squamous cell carcinoma.

Although recent studies increasingly suggest the potential anti-cancer effect of quercetin, the exact underlying mechanism remains poorly demonstrated in oral squamous cell carcinoma (oSCC). Therefore, our research explored the impacts of quercetin on the ferroptosis and mTOR/S6KP70 axis in oSCC cell lines. After treating oSCC cells with quercetin or indicated compounds and transfection with SLC7A11- or S6KP70-overexpressing plasmid, cell viability was detected by CCK-8 assay. The level of ferroptosis in oSCC cells was assessed by measuring ROS and GSH levels. The activation of mTOR/S6KP70 axis was assessed by Western blotting. Quercetin promoted ferroptosis in an mTOR/S6KP70-dependent manner to inhibit tumor growth in oSCC cells. Mechanistically, we revealed that quercetin induced lipid peroxidation and reduced GSH levels by repressing SLC7A11 expression in oSCC cells. Specifically, the effects of quercetin on ferroptosis and mTOR and S6KP70 phosphorylation were partially blocked by both mTOR agonist and S6KP70 overexpression. Moreover, mTOR inhibitor promoted ferroptosis in quercetin-treated oSCC cells. Our findings showed that ferroptosis may be a new anti-tumor mechanism of quercetin. Additionally, we identified that quercetin can target mTOR/S6KP70 cascade to inhibit the growth of oSCC cells.

Comparison of Dural Puncture Epidural, Epidural and Combined Spinal-Epidural Anesthesia for Cesarean Delivery: A Randomized Controlled Trial.

Purpose: Catheter-based techniques such as combined spinal-epidural (CSE) anesthesia which are sometimes indicated for obstetric anesthesia have a complex mechanism of action. The application of the dural puncture epidural (DPE) anesthesia for cesarean section (CS) has not been well investigated. The present study compared the relatively novel DPE technique with epidural (EA) and CSE anesthesia. Patients and Methods: We randomly assigned 150 parturients who underwent elective CS to receive DPE, EA or CSE anesthesia. The primary outcome was the onset of sensory anesthesia to the T5 dermatome assessed using the Cox proportional hazards model. Secondary outcomes included median time to sensory block, quality of block, patient and surgeon satisfaction, APGAR scores and other side effects. Results: For DPE anesthesia versus EA anesthesia, the onset of anesthesia was faster (hazard ratio 2.47 [95% CI 1.56 to 3.90], adjusted P < 0.001) and the median time to surgical level was shorter (16 [IQR 14-18] min versus 19 [15.5-21] min, adjusted P < 0.001); the incidence of intraoperative pain was lower (7/48 versus 17/47, adjusted P = 0.046) and the median patient satisfaction score was higher (9 [IQR 9-10] versus 8 [8-9.5], adjusted P = 0.004). In the CSE group, the onset of anesthesia was faster than in the other two but the incidence of hypotension was higher (P < 0.001) and the phenylephrine requirement was greater (P < 0.001). Conclusion: DPE anesthesia had a faster onset and better quality of block than EA anesthesia and provided less influence to maternal hemodynamic parameters than CSE anesthesia for CS. These results suggest that the dural puncture plays a significant role in enhancing the effectiveness of epidural top-ups during CSE anesthesia and indicates enlightenment that contributes to the satisfaction of anesthetic effect in DPE technique labor analgesia transferred to CS.

[Transport Influence and Potential Sources of PM2.5 Pollution for Nanjing].

In this study, 24-hour backward trajectories of the air mass in Nanjing were calculated by using the HYSPLIT model with the NCEP global reanalysis data from March 2019 to February 2020. The backward trajectories combined with the hourly concentration data of PM2.5 were then utilized in the trajectory clustering analysis and potential pollution source analysis. The results showed that the average concentration of PM2.5 in Nanjing was(36±20) µg·m-3 during the study period, with 17 days exceeding the grade Ⅱ national ambient air quality standards (75 µg·m-3). PM2.5 concentration exhibited clear seasonal variation, with winter (49 µg·m-3)>spring (42 µg·m-3)>autumn (31 µg·m-3)>summer (24 µg·m-3). PM2.5 concentration was significantly positively correlated with surface air pressure but significantly negatively correlated with air temperature, relative humidity, precipitation, and wind speed. Based on the trajectories, seven transport routes were identified in spring, and six routes for the other seasons. The northwest and south-southeast routes in spring, southeast route in autumn, and southwest route in winter were the main pollution transport routes in each season, with the characteristics of short transport distance and slow air mass movement, indicating that local accumulation was one of the main reasons for the high value of PM2.5 in quiet and stable weather. The distance of the northwest route in winter was large, and the PM2.5 concentration was 58 µg·m-3, which was the 2nd highest concentration in all routes, indicating that the cities in the northeast of Anhui had a great transport influence on Nanjing PM2.5. The distribution of PSCF and CWT was relatively consistent, and the main potential source areas were mainly local and adjacent areas of Nanjing, indicating that PM2.5 control is needed to strengthen local control and carry out joint prevention and control with adjacent areas. Winter was most affected by transport, its main potential source area was located at the junction of northwest Nanjing and Chuzhou, and the main source origin was in Chuzhou; therefore joint prevention and control should be expanded to Anhui.

A multiple-tissue-specific magnetic resonance imaging model for diagnosing Parkinson's disease: a brain radiomics study.

Brain radiomics can reflect the characteristics of brain pathophysiology. However, the value of T1-weighted images, quantitative susceptibility mapping, and R2* mapping in the diagnosis of Parkinson's disease (PD) was underestimated in previous studies. In this prospective study to establish a model for PD diagnosis based on brain imaging information, we collected high-resolution T1-weighted images, R2* mapping, and quantitative susceptibility imaging data from 171 patients with PD and 179 healthy controls recruited from August 2014 to August 2019. According to the inclusion time, 123 PD patients and 121 healthy controls were assigned to train the diagnostic model, while the remaining 106 subjects were assigned to the external validation dataset. We extracted 1408 radiomics features, and then used data-driven feature selection to identify informative features that were significant for discriminating patients with PD from normal controls on the training dataset. The informative features so identified were then used to construct a diagnostic model for PD. The constructed model contained 36 informative radiomics features, mainly representing abnormal subcortical iron distribution (especially in the substantia nigra), structural disorganization (e.g., in the inferior temporal, paracentral, precuneus, insula, and precentral gyri), and texture misalignment in the subcortical nuclei (e.g., caudate, globus pallidus, and thalamus). The predictive accuracy of the established model was 81.1 ± 8.0% in the training dataset. On the external validation dataset, the established model showed predictive accuracy of 78.5 ± 2.1%. In the tests of identifying early and drug-naïve PD patients from healthy controls, the accuracies of the model constructed on the same 36 informative features were 80.3 ± 7.1% and 79.1 ± 6.5%, respectively, while the accuracies were 80.4 ± 6.3% and 82.9 ± 5.8% for diagnosing middle-to-late PD and those receiving drug management, respectively. The accuracies for predicting tremor-dominant and non-tremor-dominant PD were 79.8 ± 6.9% and 79.1 ± 6.5%, respectively. In conclusion, the multiple-tissue-specific brain radiomics model constructed from magnetic resonance imaging has the ability to discriminate PD and exhibits the advantages for improving PD diagnosis.

Network of Interactions Between Gut Microbiome, Host Biomarkers, and Urine Metabolome in Carotid Atherosclerosis.

Comprehensive analyses of multi-omics data may provide insights into interactions between different biological layers concerning distinct clinical features. We integrated data on the gut microbiota, blood parameters and urine metabolites of treatment-naive individuals presenting a wide range of metabolic disease phenotypes to delineate clinically meaningful associations. Trans-omics correlation networks revealed that candidate gut microbial biomarkers and urine metabolite feature were covaried with distinct clinical phenotypes. Integration of the gut microbiome, the urine metabolome and the phenome revealed that variations in one of these three systems correlated with changes in the other two. In a specific note about clinical parameters of liver function, we identified Eubacteriumeligens, Faecalibacteriumprausnitzii and Ruminococcuslactaris to be associated with a healthy liver function, whereas Clostridium bolteae, Tyzzerellanexills, Ruminococcusgnavus, Blautiahansenii, and Atopobiumparvulum were associated with blood biomarkers for liver diseases. Variations in these microbiota features paralleled changes in specific urine metabolites. Network modeling yielded two core clusters including one large gut microbe-urine metabolite close-knit cluster and one triangular cluster composed of a gut microbe-blood-urine network, demonstrating close inter-system crosstalk especially between the gut microbiome and the urine metabolome. Distinct clinical phenotypes are manifested in both the gut microbiome and the urine metabolome, and inter-domain connectivity takes the form of high-dimensional networks. Such networks may further our understanding of complex biological systems, and may provide a basis for identifying biomarkers for diseases. Deciphering the complexity of human physiology and disease requires a holistic and trans-omics approach integrating multi-layer data sets, including the gut microbiome and profiles of biological fluids. By studying the gut microbiome on carotid atherosclerosis, we identified microbial features associated with clinical parameters, and we observed that groups of urine metabolites correlated with groups of clinical parameters. Combining the three data sets, we revealed correlations of entities across the three systems, suggesting that physiological changes are reflected in each of the omics. Our findings provided insights into the interactive network between the gut microbiome, blood clinical parameters and the urine metabolome concerning physiological variations, and showed the promise of trans-omics study for biomarker discovery.

Efficient phenol degradation by laccase immobilized on functional magnetic nanoparticles in fixed bed reactor under high-gradient magnetic field.

Enzymatic degradation of emerging contaminants has gained great interest for the past few years. However, free enzyme often incurs high costs in practice. The immobilized laccase on the polyethylenimine (PEI)-functionalized magnetic nanoparticles (Fe3O4-NH2-PEI-laccase) was fabricated to efficiently degrade phenolic compounds continuously in a newly fixed bed reactor under a high-gradient magnetic field. The degradation rate of continuous treatment in the bed after 18 h was 2.38 times as high as that of batch treatment after six successive operations with the same treatment duration. Under the optimal conditions of volume fraction of nickel wires mesh, flow rate of phenol solution, phenol concentration, and Fe3O4-NH2-PEI-laccase amount, the degradation rate of phenol kept over 70.30% in 48 h continuous treatment. The fixed bed reactor filled with Fe3O4-NH2-PEI-laccase provided a promising avenue for the continuous biodegradation of phenolic compounds for industrial wastewater in practice.

[Therapeutic effect of scalp acupuncture combined with rehabilitation training on balance dysfunction in children with spastic hemiplegia].

OBJECTIVE: To investigate the therapeutic effect of scalp acupuncture and rehabilitation training on balance dysfunction in children with spasmodic hemiplegia so as to provide the reference to the optimization of treatment scheme. METHODS: A total of 60 children with spastic hemiplegia were divided into a routine group and a scalp acupuncture group, 30 cases in each one according to random number table. In the routine group, the rehabilitation training was provided, including exercise training, balance training, spasmotherapy apparatus, electromyography biofeedback apparatus and orthoses. In the scalp acupuncture group, on the base of the treatment as the routine group, scalp acupuncture was supplemented at motor area, foot motor sensory area, equilibrium area and parietal temporal anterior oblique line. Separately, before the treatment, after 3 months treatment and after 6 months treatment, the dimension D and E of the gross motor function measure-88 (GMFM-88) and Berg balance scale (BBS) were adopted to evaluate balance related motor functions and equilibrium function. The differences in the above 3 indicators at different time stages were compared in children between the two groups. RESULTS: Compared with the score before the treatment, BBS score was obviously increased after 3 and 6 months treatment in the patients of the two groups respectively (P<0.05). The score in the dimension D and E after 6-month treatment was increased significantly as compared with the score before treatment and after 3-month treatment in the same group respectively (P<0.05). Compared with the routine group, the score of dimension D and E of GMFM-88 as well as BBS score were all increased obviously in the scalp acupuncture group after 3 and 6 months treatment (P<0.05). CONCLUSION: On the base of routine rehabilitation training, scalp acupuncture can improve balance function of children with spastic hemiplegia better.

[Effect of Baixiao moxibustion at meridian sinew nodal points on upper limb motor function in children with spastic hemiplegic cerebral palsy].

OBJECTIVE: To investigate the effect of Baixiao moxibustion at meridian sinew nodal points combined with routine rehabilitation on upper limb motor function in children with spastic hemiplegic cerebral palsy. METHODS: A total of 50 children with spastic hemiplegic cerebral palsy were divided into control group and treatment group using a random number table, with 25 children in each group. The children in the control group were given routine rehabilitation training of the ipsilateral upper and lower limbs, and those in the treatment group were given Baixiao moxibustion at the meridian sinew nodal points of the ipsilateral upper limb in addition to the treatment in the control group, once a day and five times a week. Each course of treatment was 4 consecutive weeks, and both groups were treated for 3 courses. Before treatment and at weeks 4 and 12 of treatment, modified Ashworth score was used to evaluate muscle tension of the ipsilateral upper limb, and 88-item version of the Gross Motor Function Measure (GMFM-88) and Carroll upper extremities functional test (UEFT) were used to assess the motor function of the ipsilateral upper limb. RESULTS: At weeks 4 and 12 of treatment, both groups had a significant reduction in modified Ashworth score (P<0.05) and significant increases in GMFM-88 and UEFT scores (P<0.05). Both groups had significant changes in modified Ashworth score, GMFM-88 score, and UEFT score from week 4 to week 12 of treatment (P<0.05). Compared with the control group at week 12 of treatment, the treatment group had a significant reduction in modified Ashworth score (P<0.05) and significant increases in GMFM-88 and UEFT scores (P<0.05). CONCLUSION: Baixiao moxibustion at meridian sinew nodal points can significantly improve the muscle tension and motor function of the ipsilateral upper limb in children with spastic hemiplegic cerebral palsy, and the improvement becomes more apparent as the treatment lasts longer.

T-2 toxin cytotoxicity mediated by directly perturbing mitochondria in human gastric epithelium GES-1 cells.

T-2 toxin is one of the most toxic trichothecenes and harmful to human health and animal husbandry. The mechanism underlying its growth suppression remains unclear, especially for mitochondrial damage in human gastric epithelial cells. In the present study, we investigated cell death caused by T-2 toxin in a human gastric epithelial cell line (GES-1) and the possible mechanism of T-2-induced cytotoxicity. T-2 strongly reduced the viability of GES-1 cells in a time- and dose-dependent manner within a small range of concentrations. However, when the concentrations of T-2 were >40 nM, there was no concentration dependence, only time dependence. Moreover, T-2 induced apoptosis, with the activation of caspase-3 in GES-1 and mitochondrial membrane potential (MMP) decrease and cytochrome c release. T-2 also resulted in the accumulation of reactive oxygen species (ROS) and DNA damage with a positive signal of p-H2A.X in GES-1 cells. While T-2 caused a MMP decrease, DNA damage and cell death were not blocked by pretreatment with 3 mM glutathione (GSH), a typical scavenger of ROS. The induction of mitochondrial permeability transition pore (mPTP) regulators voltage-dependent anion channel (VDAC1) and cyclophilin D (CypD) were also observed in T-2-treated cells. Interestingly, cyclosporine A (CsA), a CypD inhibitor, significantly reversed the drop in MMP and the DNA damage, as well as ROS accumulation caused by T-2. Additionally, GES-1 cell death could also be protected to some extent by 4, 4'-diisothiocyanatostilbene-2, 2'-disulfonic acid (DIDS), an inhibitor of VDAC1, especially the combination of CsA and DIDS, and 3 mM GSH could further enhance the effect of CsA + DIDS on cell viability. In conclusion, our present findings indicate that the T-2 induced MMP decrease, DNA damage and cell death, as well as ROS accumulation in GES-1 cells, starts with T-2 directly perturbing the mitochondria triggering ROS generation by acting on CypD and VDAC1. This study presents a new viewpoint for evaluating the toxicity of T-2 toxin.

[Scenario Simulation Study Constrained by the Ambient Air Quality Standards in Nanjing].

With the constraint that all six major pollutants in Nanjing must meet the air quality standards by 2030, on the basis of the 2015 emission inventory, the CMAQ air quality model was used to conduct PM2.5 sensitivity tests, and scenario analysis was used to predict the emission inventory and the air quality of four emission reduction scenarios were simulated. Finally, the total control index under the constraint of meeting the standards was obtained. The results show that primary particulate matter (PPM) reduction is the most effective at reducing the concentration of PM2.5 in the atmosphere, on the basis of emission reduction in surrounding areas, PPM emission reduction accounts for 88% of the total reduction of the annual average concentration of PM2.5, followed by NH3, NOx, SO2, and VOCs, which contribute to 10.3%, 5.5%, 3.2%, and 0.5%, respectively. Compared to 2015, the reduction ratios of the major pollutants are between 22% and 53%. Controlling the activity level is more effective for SO2, NH3 and CO emissions reduction, while there is still more opportunity for NOx and VOCs end treatment. When the emissions of SO2, NOx, PM10, PM2.5, BC, OC, CO, VOCs, and NH3 are controlled to 2.43×104, 8.47×104, 9.42×104, 3.74×104, 0.19×104, 0.30×104, 26.56×104, 13.08×104, and 1.50×104 t, respectively, it is expected that the levels of the six pollutants in Nanjing can meet the national ambient air quality level 2 standards.

Assessing inflammation in Chinese subjects with subtypes of heart failure: an observational study of the Chinese PLA Hospital Heart Failure Registry.

BACKGROUND: Inflammation is an important element of the pathophysiological process of heart failure (HF) and is correlated with subtypes of HF. The association between multiple biomarkers of inflammation and HF subtypes in Chinese subjects remains unclear. This study aimed to compare the differences in inflammation biomarkers among Chinese patients with different subtypes of HF who have been identified to date. METHODS: We included 413 consecutive patients with HF, including 262 with preserved ejection fraction (HFpEF), 55 with middle-ranged ejection fraction (HFmrEF) and 96 with reduced ejection fraction (HFrEF). Ten inflammation biomarkers were analyzed and compared according to the HF subtypes. One hundred contemporary non-HF subjects were also recruited as the control group. Moreover, the correlations between the inflammatory biomarkers and left ventricular ejection fraction of the HF subtypes were assessed. RESULTS: The mean age of the HF patients was 65.0 ± 12.0 years, 65.8% were male. Distinct subtypes of HF demonstrated different inflammation biomarker panels. IL-6, PTX-3, ANGPTL-4 and TNF-α were correlated with HFrEF; IL-1ß and PTX-3 were correlated with HFmrEF; and IL-1ß and IL-6 were correlated with HFpEF. The multivariable logistic regression showed that IL-1ß [relative ratio (RR) = 1.08, 95% CI: 1.02-1.15, P = 0.010], IL-6 (RR = 1.03, 95% CI: 1.01-1.06, P = 0.016), PTX-3 (RR = 1.31, 95% CI: 1.11-1.55, P = 0.001), and ANGPTL-4 (RR = 1.05, 95% CI: 1.02-1.07, P < 0.001) were independently associated with HF, while IL-6 (RR = 1.03, 95% CI: 1.01-1.04, P = 0.019), PTX-3 (RR = 1.23, 95% CI: 1.06-1.43, P = 0.007), and ANGPTL-4 (RR = 1.03, 95% CI: 1.01-1.06, P = 0.005) were independently associated with the HF subtype. CONCLUSIONS: Diverse inflammation biomarkers have multifaceted presentations according to the subtype of HF, which may illustrate the diverse mechanisms of inflammation in Chinese HF patients. IL-6, PTX-3, and ANGPTL-4 were independent inflammation factors associated with HFrEF and HF.

Retrospective study of immediate postoperative electron radiotherapy for therapy-resistant earlobe keloids.

Keloid resection followed by adjuvant radiotherapy is the most efficacious treatment for keloids. However, for earlobe keloids, an optimal protocol for the total dose and fractions of adjuvant radiation has not yet been established. We retrospectively analyzed the efficacy and safety of immediate three-fraction electron radiotherapy after operation for resistant earlobe keloids. From 2011 to 2017, three-fraction electron radiotherapy with single dose of 5 Gy was given postoperatively to 23 patients with 30 keloids in our hospital. The first fraction of adjuvant radiotherapy was administered within 2 h of surgery, and the other two sessions were completed within the next day or two. Five (16.7%) primary keloids and 25 (83.3%) recurrent keloids were examined in this study. The primary endpoint was the local control rate, which was 86.7% after a median follow-up of 26 months (14-93 months). Secondary endpoints were acute and late procedure-related complications, and no severe complications were observed after combination therapy. Our results suggest that three-fraction electron radiotherapy after excision within 2 days of surgery is a safe and effective protocol for the prevention of earlobe keloid recurrence that can also improve patient compliance and comfort.

Characterization and antioxidant activity of an acidic exopolysaccharide from Lactobacillus plantarum JLAU103.

An acidic exopolysaccharide (EPS) designated as EPS103 was isolated, purified and characterized from Lactobacillus plantarum JLAU103. EPS103 had a relatively lower molecular weight of 12.4 KDa and was consisted of arabinose, rhamnose, fucose, xylose, mannose, fructose, galactose, and glucose in an approximate molar ratio of 4.05: 6.04: 6.29: 5.22: 1.47: 5.21: 2.24: 1.83. A specific spectrogram of acidic polysaccharide was obtained by FT-IR analysis, and both α- and ß-type configurations were presented in EPS103 based on the nuclear magnetic resonance spectroscopy. Microstructural analysis of EPS103 demonstrated a smooth and glittering cube structure, and presence of many homogeneous rod-shaped lumps. Comprehensive study of in vitro antioxidant activity indicated that EPS103 possess strong scavenging abilities against hydroxyl, ABTS, and DPPH radicals with the maximum of 80.4%, 65.5%, and 60.5%, respectively, at 10 mg/mL concentration. Furthermore, EPS103 also showed strong ferrous ions chelating activity and oxygen radical absorbance capacity. These results together indicated that the EPS103 isolated from L. plantarum JLAU103 has great potential for use as a natural antioxidant or functional additive in foods industry.

Repeated measurement of growth-differentiation factor-15 in Chinese Han patients with post-myocardial infarction chronic heart failure.

BACKGROUND: Growth-differentiation factor-15 (GDF-15) is a promising prognostic biomarker in patients with chronic heart failure (CHF). Comparatively little is known about the value of repeated measurement of GDF-15 with CHF in Chinese Han population. This study sought to identify the clinical value of repeated measurement of GDF-15 in Chinese Han patients with post-myocardial infarction CHF. METHODS: In total, 232 consecutive Chinese Han patients with post-myocardial infarction CHF were enrolled prospectively from January 2014 to June 2016.The plasma concentration of GDF-15 was determined on admission and over 12 months. Patients were followed up for all-cause death and a composite outcome of major adverse cardiac events (MACE) included all-cause death, myocardial infarction and first heart failure (HF) re-hospitalization. Association with other clinical variables and adverse outcomes of repeated measurement of GDF-15 was explored. RESULTS: The median baseline GDF-15 level was 2025 ng/L. Baseline GDF-15 was moderately associated with baseline N-terminal pro-B type natriuretic peptide (NT-proBNP) (coefficient 0.561, P < 0.001). During a median follow-up of 20 months, there were 53 deaths and 100MACE. GDF-15 remained an independent predictor of all-cause death (adjusted hazard ratio 1.826 per 1 Ln U, 95% CI: 1.037-8.360; P = 0.037) and MACE (adjusted hazard ratio 2.243 per 1 Ln U, 95% CI: 1.181-1.775; P < 0.001) adjusted for established risk factors. Repeated measurement of GDF-15 was performed in 173 survivals over 12months. Increase of GDF-15 over 12 months was associated with dilatation of left ventricle and acted as an independent predictor of subsequent all-cause death (adjusted HR = 3.164, 95% CI: 1.245-0.041; P = 0.015). In the joint model, GDF-15 was also shown to be a risk factor for all-cause death (HR = 2.749, 95% CI: 1.667-3.831; P < 0.001) and MACE (HR = 2.434, 95% CI: 1.425-3.443; P < 0.001). CONCLUSIONS: Repeated measurements of GDF-15 have promising prognostic value of the risk of all-cause death in Chinese Han patients with CHF post-myocardial infarction. GDF-15 may influence the post-myocardial infarction CHF through the path physiological pathway of myocardial remodeling.

Ablative Hypofractionated Radiation Therapy Enhances Non-Small Cell Lung Cancer Cell Killing via Preferential Stimulation of Necroptosis In Vitro and In Vivo.

PURPOSE: To investigate how necroptosis (ie, programmed necrosis) is involved in killing of non-small cell lung cancer (NSCLC) after ablative hypofractionated radiation therapy (HFRT). METHODS AND MATERIALS: Deoxyribonucleic acid damage, DNA repair, and the death form of NSCLC cells were assessed after radiation therapy. The overexpression and silencing of receptor-interacting protein kinases 3 (RIP3, a key protein involved activation of necroptosis)-stable NSCLC cell lines were successfully constructed. The form of cell death, the number and area of colonies, and the regulatory proteins of necroptosis were characterized after radiation therapy in vitro. Finally, NSCLC xenografts and patient specimens were used to examine involvement of necroptosis after ablative HFRT in vivo. RESULTS: Radiation therapy induced expected DNA damage and repair of NSCLC cell lines, but ablative HFRT at ≥10 Gy per fraction preferentially stimulated necroptosis in NSCLC cells and xenografts with high RIP3 expression, as characterized by induction and activation of RIP3 and mixed-lineage kinase domain-like protein and release of immune-activating chemokine high-mobility group box 1. In contrast, RNA interference of RIP3 attenuated ablative HFRT-induced necroptosis and activation of its regulatory proteins. Among central early-stage NSCLC patients receiving stereotactic body radiation therapy, high expression of RIP3 was associated with improved local control and progression-free survival (all P < .05). CONCLUSIONS: Ablative HFRT at ≥10 Gy per fraction enhances killing of NSCLC with high RIP3 expression via preferential stimulation of necroptosis. RIP3 may serve as a useful biomarker to predict favorable response to stereotactic body radiation therapy.

Efficient Catalytic Oxidation of 5-Hydroxymethylfurfural to 2,5-Furandicarboxylic Acid by Magnetic Laccase Catalyst.

2,5-Furandicarboxylic acid (FDCA) is a bio-based platform chemical for the production of polyethylene furanoate (PEF) and other valuable furanic chemicals. A magnetic laccase catalyst with (2,2,6,6-tetramethyl-piperidin-1-yl)oxyl (TEMPO) as the mediator has the remarkable capability of oxidizing 5-hydroxymethylfurfural (HMF) to 2,5-furandicarboxylic acid (FDCA). Under optimal reaction conditions, a quantitative yield (90.2 %) of FDCA with complete HMF conversion was obtained after 96 h of reaction. More importantly, the magnetic laccase catalyst exhibited good recyclability and stability, maintaining 84.8 % of its original activity following six reuse cycles. This is the first report on the efficient catalytic oxidation of HMF to FDCA by using an immobilized enzyme catalyst.

Neutrophil-to-lymphocyte ratio compared to N-terminal pro-brain natriuretic peptide as a prognostic marker of adverse events in elderly patients with chronic heart failure.

BACKGROUND: The neutrophil-to-lymphocyte (N/L) ratio has been associated with poor prognosis in patients with heart failure, but it has not been compared with N-terminal pro-brain natriuretic peptide (NT-proBNP) in elderly patients with chronic heart failure (CHF). We sought to make this comparison. METHODS: A total of 1355 elderly patients with CHF were analyzed. A multivariate logistic regression model was used to analyze the variables associated with atrial fibrillation (AF). Cox regression analysis was used to assess the multivariable relationship between the N/L ratio, NT-proBNP level, and subsequent major cardiovascular events (MCE). RESULTS: In the multiple logistic regression analysis, the N/L ratio was demonstrated as a risk factor for AF in elderly patients with CHF [odds ratio (OR): 1.079, 95% confidence interval (CI): 1.027-1.134, P = 0.003]. The median follow-up period was 18 months. In a multivariable model using tertiles of both variables, the highest tertile of the N/L ratio was significantly associated with MCE [hazard ratio (HR): 1.407, 95% CI: 1.098-1.802, P = 0.007] compared with the lowest tertile. Similarly, the highest NT-proBNP tertile was also significantly associated with MCE (HR: 1.461, 95% CI: 1.104-1.934, P = 0.008). CONCLUSIONS: In elderly patients with CHF, the N/L ratio is one of the important risk factors for AF and it is an inexpensive and readily available marker with similar independent prognostic power to NT-proBNP. The risk of MCE increases 1.407-fold when the N/L ratio is elevated to the highest tertile.

Salubrinal attenuates right ventricular hypertrophy and dysfunction in hypoxic pulmonary hypertension of rats.

The phosphorylation of eukaryotic translation initiation factor 2 alpha (p-eIF2α) is essential for cell survival during hypoxia. The aim of this study was to investigate whether salubrinal, an inhibitor of p-eIF2α dephosphorylation could attenuate pulmonary arterial hypertension (PAH) and right ventricular (RV) hypertrophy in rats exposed to hypobaric hypoxia. PAH of rats was induced by hypobaric hypoxia. Salubrinal supplemented was randomized in either a prevention or a reversal protocol. At the end of the follow-up point, we measured echocardiography, hemodynamics, hematoxylin-eosin and Masson's trichrome stainings. RNA-seq analysis is explored to identify changes in gene expression associated with hypobaric hypoxia with or without salubrinal. Compared with vehicle-treatment rats exposed to hypobaric hypoxia, salubrinal prevented and partly reversed the increase of the mean pulmonary artery pressure and RV hypertrophy. What's more, salubrinal reduced the percentage wall thickness (WT%) of pulmonary artery and RV collagen volume fraction (CVF) in both prevention and reversal protocols. We also found that salubrinal was capable of reducing endoplasmic reticulum stress and oxidative stress. The result of RNA-seq analysis revealed that chronic hypoxia stimulated the differential expression of a series of genes involved in cell cycle regulation and ventricular hypertrophy and so on. Some of these genes could be ameliorated by salubrinal. These results indicate that salubrinal could prevent and reverse well-established RV remodeling, and restore the genes and pathways altered in the right ventricles of rats exposed to hypobaric hypoxia.

Genome-wide analysis of tunicamycin-induced endoplasmic reticulum stress response and the protective effect of endoplasmic reticulum inhibitors in neonatal rat cardiomyocytes.

Tunicamycin (TM) is an inducer of endoplasmic reticulum (ER) stress. However, which genes related to ER stress was induced in cardiomyocytes on a genome-wide scale remains poorly understood. Salubrinal and its derivatives are ER stress inhibitors. However, the cellular protection mechanisms remain unresolved. Neonatal rat cardiomyocytes were cultured from ventricles of one-day-old Wistar rats. Cells were exposed to salubrinal, its derivatives (PP1-12, PP1-24) or vehicle followed by TM treatment at different times. Total RNA was isolated from cells for RNA-sequencing analysis. The expressions of 189, 182, 556, 860, and 1314 genes were changed in cells exposed to TM for 1, 3, 6, 12, and 24 h. Five well-known UPR genes (Hspa5, Hsp90b1, Calr, Ddit3, and Atf4) were significantly increased in a time-dependent manner. Six not well-known genes (Hyou1, Herpud1, Manf, Creld2, Sdf2l1, and Slc3a2) were highlighted to be involved in ER stress. Compared with TM-only treated cells, the expressions of 36 genes upregulated by TM and 74 genes downregulated by TM were reversed by salubrinal. In comparison, 121 genes upregulated by TM and 92 genes downregulated by TM were reversed by PP1-12. Most genes altered by salubrinal are in the category of transcription (1 h) and cell cycle (24 h). Most genes altered by PP1-12 are in the category of response to ER stress (3 h) and cell cycle (24 h). Our findings help elucidate the mechanism for TM treatment and may be useful for future drug screens involved in ER stress.

High-content screening identifies inhibitors of the nuclear translocation of ATF6.

Activating transcription factor 6 (ATF6) is a transmembrane protein that consists of a cytoplasmic domain and an endoplasmic reticulum (ER) luminal domain. As unfolded protein levels arise in the ER, the ER cytoplasmic domain of ATF6 moves to the nucleus, where it activates the transcription of a range of genes, including those involved in apoptosis. As ATF6 only becomes functional once it has moved to the nucleus, compounds that inhibit its re-localization are of therapeutic interest. The aim of the present study was to rapidly and accurately identify such compounds using a novel image­based, high­content screening (HCS) technique. The results from the HCS analysis were then confirmed by luciferase reporter assays, western blot analysis and the measurement of cell viability. We found that HCS identified compounds which inhibited ATF6 nuclear translocation with high specificity, as confirmed by the luciferase reporter assay and western blot analysis. Moreover, we demonstrated that 3 of the 80 identified compounds impaired ATF6-mediated induced cell death. The data from this study support the theory that HCS is a novel, high throughput method which can be used for accurate and rapid compound screening.