Synergistic Antitumor Effect of Dichloroacetate in Combination with 5-Fluorouracil in Colorectal Cancer

Dichloroacetate (DCA), an inhibitor of pyruvate dehydrogenase kinase (PDK), has been recently demonstrated as a promising nontoxic antineoplastic agent that promotes apoptosis of cancer cells. In the present study, we aimed to investigate the antitumor effect of DCA combined with 5-Fluorouracil (5-FU) on colorectal cancer (CRC) cells. Four human CRC cell lines were treated with DCA or 5-FU, or a combination of DCA and 5-FU. The cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The interaction between DCA and 5-FU was evaluated by the median effect principle. Immunocytochemistry with bromodeoxyuridine (BrdU) was carried out to determine the proliferation of CRC cells. Cell cycle and apoptosis were measured by flow cytometry, and the expression of apoptosis-related molecules was assessed by western blot. Our results demonstrated that DCA inhibited the viability of CRC cells and had synergistic antiproliferation in combination with 5-FU. Moreover, compared with 5-FU alone, the apoptosis of CRC cells treated with DCA and 5-FU was enhanced and demonstrated with the changes of Bcl-2, Bax, and caspase-3 proteins. Our results suggest that DCA has a synergistic antitumor effect with 5-FU on CRC cell lines in vitro

Transcription Factor SsSte12 Was Involved in Mycelium Growth and Development in Sclerotinia sclerotiorum

Sclerotinia sclerotiorum is a challenging agricultural pathogen for management, causing large global economic losses annually. The sclerotia and infection cushions are critical for its long-term survival and successful penetration on a wide spectrum of hosts. The mitogen-activated protein kinase (MAPK) cascades serve as central signaling complexes that are involved in various aspects of sclerotia development and infection. In this study, the putative downstream transcription factor of MAPK pathway, SsSte12, was analyzed in S. sclerotiorum. Silencing SsSte12 in S. sclerotiorum resulted in phenotypes of delayed vegetative growth, reduced size of sclerotia, and fewer appressoria formation. Consequently, the SsSte12 RNAi mutants showed attenuated pathogenicity on the host plants due to the defect compound appressorium. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation assays demonstrated that the SsSte12 interacts with SsMcm1. However, the SsMcm1 expression is independent of the regulation of SsSte12 as revealed by qRT-PCR analysis in SsSte12 RNAi mutants. Together with high accumulation of SsSte12 transcripts in the early development of S. sclerotiorum, our results demonstrated that SsSte12 function was essential in the vegetative mycelial growth, sclerotia development, appressoria formation and penetration-dependent pathogenicity. Moreover, the SsSte12-SsMcm1 interaction might play a critical role in the regulation of the genes encoding these traits in S. sclerotiorum

Screening and Identification of Hub Genes in the Development of Early Diabetic Kidney Disease Based on Weighted Gene Co-Expression Network Analysis

ObjectiveThe study aimed to screen key genes in early diabetic kidney disease (DKD) and predict their biological functions and signaling pathways using bioinformatics analysis of gene chips interrelated to early DKD in the Gene Expression Omnibus database.MethodsGene chip data for early DKD was obtained from the Gene Expression Omnibus expression profile database. We analyzed differentially expressed genes (DEGs) between patients with early DKD and healthy controls using the R language. For the screened DEGs, we predicted the biological functions and relevant signaling pathways by enrichment analysis of Gene Ontology (GO) biological functions and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways. Using the STRING database and Cytoscape software, we constructed a protein interaction network to screen hub pathogenic genes. Finally, we performed immunohistochemistry on kidney specimens from the Beijing Hospital to verify the above findings.ResultsA total of 267 differential genes were obtained using GSE142025, namely, 176 upregulated and 91 downregulated genes. GO functional annotation enrichment analysis indicated that the DEGs were mainly involved in immune inflammatory response and cytokine effects. KEGG pathway analysis indicated that C-C receptor interactions and the IL-17 signaling pathway are essential for early DKD. We identified FOS, EGR1, ATF3, and JUN as hub sites of protein interactions using a protein–protein interaction network and module analysis. We performed immunohistochemistry (IHC) on five samples of early DKD and three normal samples from the Beijing Hospital to label the proteins. This demonstrated that FOS, EGR1, ATF3, and JUN in the early DKD group were significantly downregulated.ConclusionThe four hub genes FOS, EGR1, ATF3, and JUN were strongly associated with the infiltration of monocytes, M2 macrophages, and T regulatory cells in early DKD samples. We revealed that the expression of immune response or inflammatory genes was suppressed in early DKD. Meanwhile, the FOS group of low-expression genes showed that the activated biological functions included mRNA methylation, insulin receptor binding, and protein kinase A binding. These genes and pathways may serve as potential targets for treating early DKD

Comparison of the Effect of Glycemic Control in Type 2 Diabetes Outpatients Treated With Premixed and Basal Insulin Monotherapy in China

Background: Basal and premixed insulin have been widely used for insulin therapy of type 2 diabetes mellitus (T2DM) in China. The aim of this study is to compare the sustained efficacy of basal and premixed insulin therapies in T2DM outpatients with insulin monotherapy.Materials and Methods: The survey was conducted in 602 hospitals across China from April to June in 2013. The participants included outpatients who were receiving basal or premixed insulin monotherapy for more than 3 months, and the outcome was attaining a glycated hemoglobin A1C (HbA1c) of <7.0% as a measure of sustained glycemic control.Results: A total of 49,119 T2DM outpatients on basal (n = 11,967) or premixed insulin (n = 37,152) monotherapy were included in the final analyses. Using multivariable model analysis, patients using premixed insulin exhibited a better glycemic control, with more outpatients achieving the target HbA1c level than those using basal insulin (model 1, OR 0.695, 95%CI 0.664–0.728; model 2, OR 0.708, 95%CI 0.676–0.742; model 3, OR 0.717, 95%CI 0.684–0.752; model 4, OR 0.750, 95%CI 0.715–0.787). Using subgroup analysis stratified by age, sex, duration of diabetes, duration of insulin treatment, and complications, still more outpatients in every subgroup treated with premixed insulin achieved the target HbA1c (HbA1c < 7%) than those receiving basal insulin.Conclusions: Premixed insulin monotherapy had a better glycemic control (HbA1c < 7.0%) than basal insulin monotherapy for Chinese T2DM outpatients in daily

Optimising two-stage vacuum heat treatment for a high-strength micro-alloyed steel in railway spring clip application: impact on microstructure and mechanical performance

The heat treatment process is a vital step for manufacturing high-speed railway spring fasteners. In this study, orthogonal experiments were carried out to obtain reliable optimised heat treatment parameters through a streamlined number of experiments. Results revealed that a better comprehensive mechanical performance could be obtained under the following combination of heat treatment parameters: quenching temperature of 850 °C, holding time of 35 min, medium of 12% polyalkylene glycol (PAG) aqueous solution, tempering temperature of 460 °C, and holding time of 60 min. As one of the most important testing criteria, fatigue performance would be improved with increasing strength. Additionally, a high ratio of martensite to ferrite is proven to improve the fatigue limit more significantly. After this heat treatment process, the metallographic microstructure and mechanical properties satisfy the technical requirements for the high-speed railway practical operation. These findings provide a valuable reference for the practical forming process of spring fasteners

GWAS Analysis and QTL Identification of Fiber Quality Traits and Yield Components in Upland Cotton Using Enriched High-Density SNP Markers

It is of great importance to identify quantitative trait loci (QTL) controlling fiber quality traits and yield components for future marker-assisted selection (MAS) and candidate gene function identifications. In this study, two kinds of traits in 231 F6:8 recombinant inbred lines (RILs), derived from an intraspecific cross between Xinluzao24, a cultivar with elite fiber quality, and Lumianyan28, a cultivar with wide adaptability and high yield potential, were measured in nine environments. This RIL population was genotyped by 122 SSR and 4729 SNP markers, which were also used to construct the genetic map. The map covered 2477.99 cM of hirsutum genome, with an average marker interval of 0.51 cM between adjacent markers. As a result, a total of 134 QTLs for fiber quality traits and 122 QTLs for yield components were detected, with 2.18–24.45 and 1.68–28.27% proportions of the phenotypic variance explained by each QTL, respectively. Among these QTLs, 57 were detected in at least two environments, named stable QTLs. A total of 209 and 139 quantitative trait nucleotides (QTNs) were associated with fiber quality traits and yield components by four multilocus genome-wide association studies methods, respectively. Among these QTNs, 74 were detected by at least two algorithms or in two environments. The candidate genes harbored by 57 stable QTLs were compared with the ones associated with QTN, and 35 common candidate genes were found. Among these common candidate genes, four were possibly “pleiotropic.” This study provided important information for MAS and candidate gene functional studies

The influence of negative pressure during gas extraction in low-permeability coal seams on the effect of gas extraction

In order to improve the extraction method of gas extraction in coal seams with low permeability, the quadratic polynomial formula between the amount of extracted pure gas and negative pressure during gas extraction (NPGE) can be deduced based on the basic theory of oil and gas seepage mechanics in porous media and the pressure function theory of distribution of gas pressure. By using the measurement of orifice flowmeter, the formula of the amount of extracted mixed gas can be acquired and then the expression between the gas extraction concentration (GEC) and NPGE is deduced. Thus, the equation set concerning gas extraction effect is composed of three equations: the equation between the amount of extracted pure gas and NPGE, the formula for calculating the amount of extracted mixed gas, and the formula of relationship between the GEC and NPGE. According to the equation set, the GEC and the amount of extracted pure gas are calculated to further draw the scatter diagram and trend line of function. Afterwards, by analyzing the calculation result and trend line of functions, it can be seen that the increase amplitude of GEC is limited by improving the NPGE. By observing the on-site NPGE of the 20109 Wangjialing Coal mine (Shanxi Province, China), it can be speculated that with increasing NPGE, the increase trends of the GEC and the amount of extracted pure gas were consistent with that deduced theoretical formula for calculating the amount of extracted pure gas. It was basically shown as a quadratic polynomial relation in which the amount of extracted pure gas insignificantly increased. Therefore, it is infeasible to improve the gas extraction in coal seams with low permeability only relying on increasing the NPGE, and other technology methods are necessary

Numerical simulation on reasonable hole-sealing depth of boreholes for gas extraction

To overcome the low efficiency of extracting gas in coal reservoirs with a low gas permeability, some boreholes were drilled for gas extraction in No. 2 coal reservoir of Wangjialing Coalmine in Shanxi Province, China and reasonably sealed. Aiming at shortfalls such as rapid attenuation of volume for extracted gas as well as low gas permeability when using boreholes in the No. 2 coal reservoir, the traditional COMSOL MultiphysicsMT Earth Science Module was used to couple the three governing equations (Darcy-Brinkman–Navier-Stokes) for fluids. On this basis, numerical simulation on the seepage law along the directions of roadways and boreholes was carried out. The simulation results indicated that when the hole-sealing length was within the width range of fractures in roadways, the negative pressure not only led the gas in surrounding rock masses to flow to the boreholes, but also made the air flow in roadways to permeate into coal walls. As a result, gas and air flows both entered into the boreholes through the loosening zone containing fractures, resulting in seepage of air in roadway to the boreholes. The seepage velocity along the roadway direction under condition with a hole-sealing length of 12 m was obviously slower than that when the hole-sealing length was 8 m. While, the method by simply increasing the length of the hole-sealing section for boreholes failed to effectively stop the air flow in roadways from permeating into the coal wall and then entering the boreholes. Moreover, the increase in the hole-sealing length brought about much more difficulties to the hole-sealing construction. So, the method is not operable in practical condition of the coal mine. Therefore, it is necessary to improve the traditional hole-sealing technology based on foamed macromolecular materials which are mainly made of polyurethane (PU) and use the fluid wall-type hole-sealing technology based on solid-liquid coupling. Then, the effects of gas extraction before and after using the fluid wall-type hole-sealing technology based on solid-liquid coupling to increase the hole-sealing length to 12 m were compared. The comparison results revealed that the pure extraction amount of gas from a single borehole in the No. 2 coal reservoir of Wangjialing Coalmine was improved by 4∼6 times. In addition, the concentration of extracted gas increased from less than 1% under the traditional hole-sealing mode to 20%∼25%, with an increase of more than 20 times. The extraction effect of the No. 2 coal reservoir of the coal mine was significantly enhanced by employing the fluid-wall-type hole-sealing technology based on solid-liquid coupling