siRNA directed against c-Myc inhibits proliferation and downregulates human telomerase reverse transcriptase in human colon cancer Colo 320 cells

The c-Myc and human telomerase reverse transcriptase gene (hTERT) gene are frequently deregulated and overexpressed in malignancy. hTERT activity is induced by c-Myc and strategies designed to inhibit c-Myc expression in cancer cells may have considerable therapeutic value. We designed and used a short hairpin RNA to inhibit c-Myc expression in Colo 320 cells and validated its effect on cell proliferation. In this study, four c-Myc-shRNA expression vectors were constructed and introduced into Colo 320 cells. The effects of c-Myc silencing on tumor cell growth was assessed by soft agar assay and DNA synthesis experiments. The expressions of c-Myc and hTERT were also assessed by real-time reverse transcription-polymerase chain reaction and Western blot analysis. Upon transient transfection with plasmid encoding shRNA, it was found that expression of c-Myc and hTERT decreased in shRNA-transfected cells. The downregulation of c-Myc and hTERT inhibited cell growth, shortened telomere lengths, and suppressed telomerase activity. In conclusion, our findings demonstrate that shRNA of c-Myc can inhibit the DNA replication in Colo 320 cells effectively and reduce telomere length and telomerase activity, therefore, it could be used as a new potential anticancer tool for therapy of human colon cancer

FEDSM2013-16598 NUMERICAL OPTIMAL DESIGN OF IMPELLER BACK PUMP-OUT VANES ON AXIAL THRUST IN CENTRIFUGAL PUMPS

ABSTRACT Axial thrust in centrifugal pu mps attracts extensive attention in order to improve the operating reliability of pu mps. High axial thrust can cause rapid thrust bearing wear and subsequent pump failure o r frequent overhauls. A centrifugal pump (XA65/20) was selected in this study, based on L 16 (4 3 ) orthogonal array and CFD methods. The time -averaged Navier-Stokes equation was calcu lated for a 3D steady flow in the model pump in ANSYS CFX with the standard k-ω turbulence model and standard wall function applied. The structured meshes with different numbers were used for comparison in order to confirm that the computational results were not influenced by the mesh. Meanwhile, the effects of impeller back pump -out vane geometrical parameters, including its thickness S k , its outlet diameter D e and axial clearance δ, on the axial thrust and performances of the model centrifugal pu mp were analy zed. The different orthogonal schemes were obtained on the different values of S k , D e , and δ. Finally, when the parameters of the impeller S k , D e , and δ are 5mm, 100mm, 1.5mm, respectively. The Best Efficiency Point (BEF) of 69.9% was achieved with 60.12m for the designed head and -952.133N for the minimu m total axial force. The corresponding impeller with minimu m total axial fo rce was considered as the optimal scheme and manufactured for experimental test. The external characteristics by CFD have a good agreement with their experimental data, wh ich also better verified the accuracy of the numerical method of axial thrust applied in this research. Back pump-out vane thickness Back pump-out vane outlet diameter Z Back pump-out vane number N (r/min) Rotating speed ρ (kg/m3) Density g (m/s2) Gravity acceleration H (m) Pressure head ω (rad/s) Angular speed p (Pa) Pressure INTRODUCTION One of the most challenging aspects in horizontal pumps design is represented by the accurate evaluation of the axial thrust acting on the rotating shaft. In order to balance axial thrust of centrifugal pu mps, many devices such as balancing disk, balancing dru m, balancing hole and sealing system are used In this paper, the model pu mp of XA65/20 was designed to study its axial thrust and external characteristics with S k , δ an

The Gaps Between Current Management of Intracerebral Hemorrhage and Evidence-Based Practice Guidelines in Beijing, China

Background: The leading cause of death in China is stroke, a condition that also contributes heavily to the disease burden. Nontraumatic intracerebral hemorrhage (ICH) is the second most common cause of stroke. Compared to Western countries, in China the proportion of ICH is significantly higher. Standardized treatment based on evidence-based medicine can help reduce ICH's burden. In the present study we aimed to explore the agreement between the management strategies during ICH's acute phase and Class I recommendations in current international practice guidelines in Beijing (China), and to elucidate the reasons underlying any inconsistencies found.Method: We retrospectively collected in-hospital data from 1,355 ICH patients from 15 hospitals in Beijing between January and December 2012. Furthermore, a total of 75 standardized questionnaires focusing on ICH's clinical management were distributed to 15 cooperative hospitals. Each hospital randomly selected five doctors responsible for treating ICH patients to complete the questionnaires.Results: Numerous approaches were in line with Class I recommendations, as follows: upon admission, all patients underwent radiographic examination, about 93% of the survivors received health education and 84.5% of those diagnosed with hypertension were prescribed antihypertensive treatment at discharge, in-hospital antiepileptic drugs were administered to 91.8% of the patients presenting with seizures, and continuous monitoring was performed for 88% of the patients with hyperglycemia on admission. However, several aspects were inconsistent with the guidelines, as follows: only 14.2% of the patients were initially managed in the neurological intensive care unit and 22.3% of the bedridden patients received preventive treatment for deep vein thrombosis (DVT) within 48 h after onset. The questionnaire results showed that imaging examination, blood glucose monitoring, and secondary prevention of ICH were useful to more clinicians. However, the opposite occurred for the neurological intensive care unit requirement. Regarding the guidelines' recognition, no significant differences among the 3 education subgroups were observed (p > 0.05).Conclusions: Doctors have recognized most of ICH's evidence-based practice guidelines. However, there are still large gaps between the management of ICH and the evidence-based practice guidelines in Beijing (China). Retraining doctors is required, including focusing on preventing DVT providing a value from the National Institutes of Health Stroke Scale and Glasgow Coma Scalescores at the time of admission

Exploring the shared molecular mechanism of microvascular and macrovascular complications in diabetes: Seeking the hub of circulatory system injury

BackgroundMicrovascular complications, such as diabetic retinopathy (DR) and diabetic nephropathy (DN), and macrovascular complications, referring to atherosclerosis (AS), are the main complications of diabetes. Blindness or fatal microvascular diseases are considered to be identified earlier than fatal macrovascular complications. Exploring the intrinsic relationship between microvascular and macrovascular complications and the hub of pathogenesis is of vital importance for prolonging the life span of patients with diabetes and improving the quality of life.Materials and methodsThe expression profiles of GSE28829, GSE30529, GSE146615 and GSE134998 were downloaded from the Gene Expression Omnibus database, which contained 29 atherosclerotic plaque samples, including 16 AS samples and 13 normal controls; 22 renal glomeruli and tubules samples from diabetes nephropathy including 12 DN samples and 10 normal controls; 73 lymphoblastoid cell line samples, including 52 DR samples and 21 normal controls. The microarray datasets were consolidated and DEGs were acquired and further analyzed by bioinformatics techniques including GSEA analysis, GO-KEGG functional clustering by R (version 4.0.5), PPI analysis by Cytoscape (version 3.8.2) and String database, miRNA analysis by Diana database, and hub genes analysis by Metascape database. The drug sensitivity of characteristic DEGs was analyzed.ResultA total of 3709, 4185 and 8086 DEGs were recognized in AS, DN, DR, respectively, with 1820, 1666, 888 upregulated and 1889, 2519, 7198 downregulated. GO and KEGG pathway analyses of DEGs and GSEA analysis of common differential genes demonstrated that these significant sites focused primarily on inflammation-oxidative stress and immune regulation pathways. PPI networks show the connection and regulation on top-250 significant sites of AS, DN, DR. MiRNA analysis explored the non-coding RNA upstream regulation network and significant pathway in AS, DN, DR. The joint analysis of multiple diseases shows the common influenced pathways of AS, DN, DR and explored the interaction between top-1000 DEGs at the same time.ConclusionIn the microvascular and macrovascular complications of diabetes, immune-mediated inflammatory response, chronic inflammation caused by endothelial cell activation and oxidative stress are the three links linking atherosclerosis, diabetes retinopathy and diabetes nephropathy together. Our study has clarified the intrinsic relationship and common tissue damage mechanism of microcirculation and circulatory system complications in diabetes, and explored the mechanism center of these two vascular complications. It has far-reaching clinical and social value for reducing the incidence of fatal events and early controlling the progress of disabling and fatal circulatory complications in diabetes

CatNorth: An Improved Gaia DR3 Quasar Candidate Catalog with Pan-STARRS1 and CatWISE

A complete and pure sample of quasars with accurate redshifts is crucial for quasar studies and cosmology. In this paper, we present CatNorth, an improved Gaia DR3 quasar candidate catalog with more than 1.5 million sources in the 3$\pi$ sky built with data from Gaia, Pan-STARRS1, and CatWISE2020. The XGBoost algorithm is used to reclassify the original Gaia DR3 quasar candidates as stars, galaxies, and quasars. To construct training/validation datasets for the classification, we carefully built two different master stellar samples in addition to the spectroscopic galaxy and quasar samples. An ensemble classification model is obtained by averaging two XGBoost classifiers trained with different master stellar samples. Using a probability threshold of $p_{\mathrm{QSO\_mean}}>0.95$ in our ensemble classification model and an additional cut on the logarithmic probability density of zero proper motion, we retrieved 1,545,514 reliable quasar candidates from the parent Gaia DR3 quasar candidate catalog. We provide photometric redshifts for all candidates with an ensemble regression model. For a subset of 89,100 candidates, accurate spectroscopic redshifts are estimated with the Convolutional Neural Network from the Gaia BP/RP spectra. The CatNorth catalog has a high purity of > 90% while maintaining high completeness, which is an ideal sample to understand the quasar population and its statistical properties. The CatNorth catalog is used as the main source of input catalog for the LAMOST phase III quasar survey, which is expected to build a highly complete sample of bright quasars with $i < 19.5$.Comment: 24 pages, 13 figures, submitted to AAS journals. Table 4 (The CatNorth quasar candidate catalog) is available at https://nadc.china-vo.org/res/r101313

Regulation of Wnt Singaling Pathway by Poly (ADP-Ribose) Glycohydrolase (PARG) Silencing Suppresses Lung Cancer in Mice Induced by Benzo(a)pyrene Inhalation Exposure

Benzo(a)pyrene (BaP) is a polycyclic aromatic hydrocarbon that specifically causes cancer and is widely distributed in the environment. Poly (ADP-ribosylation), as a key post-translational modification in BaP-induced carcinogenesis, is mainly catalyzed by poly (ADP-ribose) glycohydrolase (PARG) in eukaryotic organisms. Previously, it is found that PARG silencing can counteract BaP-induced carcinogenesis in vitro, but the mechanism remained unclear. In this study, we further examined this process in vivo by using heterozygous PARG knockout mice (PARG+/−). Wild-type and PARG+/− mice were individually treated with 0 or 10 μg/m3 BaP for 90 or 180 days by dynamic inhalation exposure. Pathological analysis of lung tissues showed that, with extended exposure time, carcinogenesis and injury in the lungs of WT mice was progressively worse; however, the injury was minimal and carcinogenesis was not detected in the lungs of PARG+/− mice. These results indicate that PARG gene silencing protects mice against lung cancer induced by BaP inhalation exposure. Furthermore, as the exposure time was extended, the protein phosphorylation level was down-regulated in WT mice, but up-regulated in PARG+/− mice. The relative expression of Wnt2b and Wnt5b mRNA in WT mice were significantly higher than those in the control group, but there was no significant difference in PARG+/− mice. Meanwhile, the relative expression of Wnt2b and Wnt5b proteins, as assessed by immunohistochemistry and Western blot analysis, was significantly up-regulated by BaP in WT mice; while in PARG+/− mice it was not statistically affected. Our work provides initial evidence that PARG silencing suppresses BaP induced lung cancer and stabilizes the expression of Wnt ligands, PARG gene and Wnt ligands may provide new options for the diagnosis and treatment of lung cancer

Stemness Analysis Uncovers That The Peroxisome Proliferator-Activated Receptor Signaling Pathway Can Mediate Fatty Acid Homeostasis In Sorafenib-Resistant Hepatocellular Carcinoma Cells

Hepatocellular carcinoma (HCC) stem cells are regarded as an important part of individualized HCC treatment and sorafenib resistance. However, there is lacking systematic assessment of stem-like indices and associations with a response of sorafenib in HCC. Our study thus aimed to evaluate the status of tumor dedifferentiation for HCC and further identify the regulatory mechanisms under the condition of resistance to sorafenib. Datasets of HCC, including messenger RNAs (mRNAs) expression, somatic mutation, and clinical information were collected. The mRNA expression-based stemness index (mRNAsi), which can represent degrees of dedifferentiation of HCC samples, was calculated to predict drug response of sorafenib therapy and prognosis. Next, unsupervised cluster analysis was conducted to distinguish mRNAsi-based subgroups, and gene/geneset functional enrichment analysis was employed to identify key sorafenib resistance-related pathways. In addition, we analyzed and confirmed the regulation of key genes discovered in this study by combining other omics data. Finally, Luciferase reporter assays were performed to validate their regulation. Our study demonstrated that the stemness index obtained from transcriptomic is a promising biomarker to predict the response of sorafenib therapy and the prognosis in HCC. We revealed the peroxisome proliferator-activated receptor signaling pathway (the PPAR signaling pathway), related to fatty acid biosynthesis, that was a potential sorafenib resistance pathway that had not been reported before. By analyzing the core regulatory genes of the PPAR signaling pathway, we identified four candidate target genes, retinoid X receptor beta (RXRB), nuclear receptor subfamily 1 group H member 3 (NR1H3), cytochrome P450 family 8 subfamily B member 1 (CYP8B1) and stearoyl-CoA desaturase (SCD), as a signature to distinguish the response of sorafenib. We proposed and validated that the RXRB and NR1H3 could directly regulate NR1H3 and SCD, respectively. Our results suggest that the combined use of SCD inhibitors and sorafenib may be a promising therapeutic approach

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

A Review on Hydrodynamic Performance and Design of Pump-Jet: Advances, Challenges and Prospects

A pump-jet, which is generally and widely adopted on underwater vehicles for applications from deep sea exploration to mine clearing, consists of a rotor, stator, and duct, with the properties of high critical speed, high propulsion efficiency, great anti-cavitation performance, and low radiated noise. The complex interaction of the flow field between the various components and the high degree of coupling with the appendage result in the requirements of in-depth research on the hydrodynamic performance and flow field for application and design. Due to the initial application on the military field and complicated structure, there is scant literature in the evaluation of pump-jet performance and optimal design. This paper, in a comprehensive and specialized way, summarizes the pump-jet hydrodynamic performance, noise performance, and flow field characteristics involving cavitation erosion and vortices properties of tip-clearance, the interaction between the rotor and the stator and the wake field, as well as the optimal design of the pump-jet. The merits and applications range of numerical and experimental methods are overviewed as well as the design method. It also concludes the main challenges faced in practical applications and proposes a vision for future research. It was found that the compact structure and complex internal and external flow field make the pump-jet significantly different, also leading to higher performance. As the focus of cavitation research, vortices interact with the complex structure of the pump-jet, leading to instabilities of the flow field, such as vibration, radiated noise, and cavitation erosion. The effective approaches are adopted to reduce radiated pump-jet with minimal influence on the hydrodynamic performance, such as eliminating the tip clearance and installing the sawtooth duct. Advanced optimal technology can achieve high performance, cavitation performance, and acoustic performance, possessing good prospects. Further developments in investigation and the application of pump-jets in the multidisciplinary integration of fluid dynamics, acoustics, materials, chemistry, and bionics should be the main focus in future research