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Objective. This study aimed to uncover biologically significant RNAs in nucleus pulposus tissues of human intervertebral disc degeneration (IVDD) by integrated transcriptional profiling. Methods. From the Gene Expression Omnibus (GEO) database, three IVDD-related microarray profiling datasets were retrieved and assessed by intragroup data repeatability test. Then, differentially expressed circRNAs, lncRNAs, mRNAs, and miRNAs were screened in nucleus pulposus tissues between IVDD and control samples via the limma package. Coexpression networks were separately conducted via weighted gene correlation network analysis (WGCNA). Based on the feature RNAs in the IVDD-related modules, IVDD-related circRNA-miRNA-mRNA and lncRNA-miRNA-mRNA networks were conducted. The differentially expressed mRNAs in the two networks were analyzed by protein-protein interaction (PPI) and functional enrichment analyses. Results. By the intragroup data repeatability test, outlier samples were removed. Abnormally expressed RNAs were separately identified in nucleus pulposus between IVDD and controls. Via WGCNA, IVDD-related coexpression modules were constructed and the feature circRNAs, lncRNAs, mRNAs, and miRNAs were identified. Then, the circRNA- and lncRNA-miRNA-mRNA networks were built for IVDD. These mRNAs in the network exhibited complex interactions. Moreover, they were involved in distinct IVDD-related biological processes and pathways such as transcription, cell proliferation, TNF, TGF-β, and HIF signaling pathways. Conclusion. This study revealed biologically significant noncoding RNAs and their complex regulatory networks for IVDD.

BackgroundPrimary Sjögren’s syndrome (pSS) is a systemic autoimmune disease characterised by aberrant B cell hyperactivation, whose mechanism is partially understood.MethodsWe performed whole transcriptome sequencing of B cells from three pSS patients and three matched healthy controls (HC). Differentially expression genes (DEGs) were confirmed with B cells from 40 pSS patients and 40 HC by quantitative PCR and western blot. We measured the proliferation potential and immunoglobulins production of siRNA-transfected or plasmid-transfected B cells stimulated with cytosine-phosphate-guanine (CpG) or anti-IgM. We also explored Toll-like receptor 9 (TLR9) signalling to reveal the potential mechanism of B cell hyperactivation in pSS.ResultsWe identified 77 upregulated and 32 downregulated DEGs in pSS B cells. We confirmed that epithelial stromal interaction (EPST1) expression in pSS B cells was significantly higher than that from HCs. EPSTI1-silencing B cells stimulated with CpG were less proliferated and produced lower level of IgG and IgM comparing with control B cells. EPSTI1-silencing B cells expressed lower level of p-p65 and higher level of IκBα, and B cells with overexpressed EPSTI1 showed higher level of p-p65 and lower level of IκBα. Finally, IκBα degradation inhibitor Dehydrocostus Lactone treatment attenuated p65 phosphorylation promoted by EPSTI1.ConclusionElevated EPSTI1 expression in pSS B cells promoted TLR9 signalling activation and contributed to the abnormal B cell activation, which was promoted by facilitating p65 phosphorylation and activation of NF-κB signalling via promoting IκBα degradation. EPSTI1 might be implicated in pSS pathogenesis and was a potential therapeutic target of pSS.

Triacylglycerol lipase (TGL) is an essential lipid metabolism enzyme that also plays a critical role in energy metabolism; however, how it regulates other life processes is unknown. To investigate the functional role of TGL in moth reproduction, males Sitotroga cerealella were used as a model. The TGL gene was cloned and analysed. The results showed that the open reading frame of TGL was 1968 bp long and contained three conserved regions. TGL gene expression was higher in the larval and early adult stages than in the pupal stage, with the highest levels observed in the fat body, testis and accessory glands during the early adult stage. Moreover, after TGL in male adults was silenced through RNAi, the protein content in male accessory glands remained unchanged, and the spermatophore transferred into females mated with TGL -silenced males became small and empty; meanwhile, the number of apyrene sperm in the spermatophore was significantly reduced due to the reduction of apyrene sperm in males, which eventually led to the significant reduction of egg laying amount. All of the findings suggest that TGL regulates the amount of sperm in male moths as well as the morphology and quality of spermatophores transferred to females after mating with treated males, implying that TGL is critical for Sitotroga cerealella’s reproductive process.

Background Propofol (PPF) ameliorates ischemia/reperfusion (I/R) injury in multiple organs by reducing apoptosis and release of pro-inflammatory cytokines. This study aims to explore the mechanism of PPF in attenuating myocardial ischemia-reperfusion injury (MIRI). Materials and Methods Rat MIRI model was established, and PPF pre-treatment was performed 10 min before I/R. H9c2 cardiomyocytes treated with hypoxia/reoxygenation (H/R) were used to establish an in vitro model. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to evaluate HOTAIR and miR-17-5p expression levels. Flow cytometry was employed to detect the apoptosis of H9c2 cells. The interaction between HOTAIR and miR-17-5p was determined by bioinformatics analysis, luciferase reporter gene analysis, and RNA immunoprecipitation experiments. STAT3 and p-STAT3 expressions were detected by Western blot. Results PPF pre-treatment significantly reduced creatine kinase isoenzyme (CK-MB) and serum lactate dehydrogenase (LDH) levels in the serum of the rats with MIRI. PPF pre-treatment remarkably up-regulated HOTAIR expression and down-regulated miR-17-5p expression in both in vivo and in vitro models. HOTAIR adsorbed miR-17-5p to repress the expression of miR-17-5p. PPF pre-treatment markedly inhibited cardiomyocyte apoptosis induced by I/R or H/R. HOTAIR knockdown could partially reverse the protective effects of PPF on MIRI. HOTAIR could activate STAT3 signaling via repressing miR-17-5p expression. Conclusion PPF protects the MIRI by modulating the HOTAIR/miR-17-5p/STAT3 axis.

During mouse oocyte meiotic maturation, actin filaments play multiple roles in meiosis such as spindle migration and cytokinesis. FASCIN is shown to be an actin-binding and bundling protein, making actin filaments tightly packed and parallel-aligned, and FASCIN is involved in several cellular processes like adhesion and migration. FASCIN is also a potential prognostic biomarker and therapeutic target for the treatment of metastatic disease. However, little is known about the functions of FASCIN in oocyte meiosis. In the present study, we knocked down the expression of FASCIN, and our results showed that FASCIN was essential for oocyte maturation. FASCIN was all expressed in the different stages of oocyte meiosis, and it mainly localized at the cortex of oocytes from the GV stage to the MII stage and showed a similar localization pattern with actin and DAAM1. Depletion of FASCIN affected the extrusion of the first polar body, and we also observed that some oocytes extruded from the large polar bodies. This might have resulted from the defects of actin assembly, which further affected the meiotic spindle positioning. In addition, we showed that inhibition of PKC activity decreased FASCIN expression, indicating that FASCIN might be regulated by PKC. Taken together, our results provided evidence for the important role of FASCIN on actin filaments for spindle migration and polar body extrusion in mouse oocyte meiosis.

Aim and Objective: Integrating multi-omics data to identify driver genes and key biological functions for tumorigenesis remains a major challenge. Method: A new computational pipeline was developed to identify the Driver Mutation-Differential Co-Expression (DM-DCE) modules based on dysfunctional networks across 11 TCGA cancers. Results: Functional analyses provided insight into the properties of various cancers, and found common cellular signals / pathways of cancers. Furthermore, the corresponding network analysis identified conservations or interactions across different types of cancers, thus the crosstalk between the key signaling pathways, immunity and cancers was found. Clinical analysis also identified key prognostic / survival patterns. Conclusion: Taken together, our study sheds light on both cancer-specific and cross-cancer characteristics systematically.

The intestinal microecological environment is critical to an infant's growth. For those infants consuming milk power, it is very important to improve the intestinal microecological environment to promote the healthy growth of infants. In this paper, Milk protein hydrolysate (MPH), consisting of different proportions of proteins and small molecule peptides (5:5, 4:6, 3:7, 2:8, 1:9) were added to infant formula powder (IFP). The effects of MFP-enriched IFP addition on proliferation and metabolism of Bifidobacterium L80 were studied. Compared with MPH-free IFP, MFP-enriched IFP with 1:9 of proteins to small molecule peptides significantly enhanced the proliferation of Bifidobacterium L80, resulting in higher cell density, greater viable counts and higher titratable acidity. MFP-enriched IFP increased the content of seven organic acids and H2O2 in the system, and improved the antibacterial activity to E. coli BL21. This study suggested that MPH could be an effective addition to infant formula powder to promote the growth of Bifidobacterium, so to improve the intestinal health of infants.

This study investigated the effects of spontaneous and Lactobacillus plantarum (LP) fermentation on structure glutinous proso millet starch (GPM) and physical properties. In order to explore starch changes by fermentation for quality regulation of fermented food glutinous proso millet and improve properties of modified GPM starch products. The molecular weight, amylopectin chain length distribution, morphological properties, short-range ordered structure, and crystal structure of GPM starch were analyzed. Results showed that compared with the non-fermented GPM starch, amylose content of spontaneous and LP-fermented GPM starch increased then decreased, with the latter decreasing faster. There were no changes in the Type A structures of spontaneous fermented, LP-fermented, and non-fermented GPM starch. After 24 h of LP fermentation, higher water swelling power, and lower GPM starch solubility were obtained compared to non-fermentation and spontaneous fermentation. The decrease in pasting properties and thermal properties while increasing GPM starch properties was shown after fermentation compared to non-fermentation experiments. Besides, LP fermentation showed faster changes of these properties than spontaneous fermentation. In conclusion, our findings showed that fermentation could improve GPM starch properties, benefiting fermented GPM products and as a modification method for GPM starch. Especially, LP fermentation was more effective than spontaneous fermentation.

Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) have emerged as important mediators of intercellular communication in response to cartilage damage. In this study, we sought to characterize the inhibitory role of microRNA (miR)-31 encapsulated in synovial MSC (SMSC)-derived EVs in knee osteoarthritis (OA). The expression of miR-31, lysine demethylase 2A (KDM2A), E2F transcription factor 1 (E2F1), and pituitary tumor transforming gene 1 (PTTG1) was validated in cartilage tissues of knee OA patients. Following SMSC-EV extraction and identification, chondrocytes with the miR-31 inhibitor were added with SMSC-EVs, whereupon the effects of miR-31 on proliferation and migration of chondrocytes were assessed. The interaction among miR-31, KDM2A, E2F1, and PTTG1 in chondrocyte activities was probed in vitro, along with an in vivo mouse knee OA model. We identified downregulated miR-31, E2F1, and PTTG1 and upregulated KDM2A in cartilage tissues of knee OA patients. SMSC-EV-packaged miR-31 potentiated chondrocyte proliferation and migration as well as cartilage formation by targeting KDM2A. Mechanistically, KDM2A bound to the transcription factor E2F1 and inhibited its transcriptional activity. Enrichment of E2F1 in the PTTG1 promoter region activated PTTG1 transcription, accelerating chondrocyte proliferation and migration. SMSC-EVs and EVs from miR-31-overexpressed SMSCs alleviated cartilage damage and inflammation in knee joints in vivo. SMSC-EV-encapsulated miR-31 ameliorates knee OA via the KDM2A/E2F1/PTTG1 axis., Graphical Abstract, With the purpose of identifying the role of SMSC-secreted EVs containing miR-31 in knee OA, clinical samples, commercially purchased cell lines, and animals were used in a series of in vivo and in vitro experiments. The experimental data uncovered that SMSC-EV-encapsulated miR-31 ameliorates knee OA via the KDM2A/E2F1/PTTG1 axis.

In this study, the mineral-weathering bacterium Pseudomonas azotoformans F77, which was isolated from the soil of a debris flow area, was evaluated for its weathering activity under direct contact with biotite or without contact. Then, biotite-weathering behaviors of strain F77, mutants that had been created by deleting the gcd and adh genes (which are involved in gluconic acid metabolism and pilus formation, respectively), and the double mutant F77ΔgcdΔadh were compared. The relative gene expression levels of F77 and its mutants F77Δgcd and F77Δadh were also analyzed in the presence of biotite. Direct contact with biotite increased Fe and Al release from the mineral in the presence of F77. All strains had similar abilities to release Fe and Al from the mineral except for F77Δgcd and F77Δadh. Mobilized Fe and Al concentrations were decreased by up to 72, 26, and 87% in the presence of F77Δgcd, F77Δadh, and F77ΔgcdΔadh, respectively, compared to levels observed in the presence of F77 during the mineral-weathering process. Gluconic acid production was decreased for F77Δgcd and F77ΔgcdΔadh, while decreased cell attachment on the mineral surface was observed for F77Δadh, compared to findings for F77. The F77 genes involved in pilus formation and gluconic acid metabolism showed increased expression levels in the presence of biotite. The results of this study showed important roles for the genes involved in gluconic acid metabolism and pilus formation in mineral weathering by F77 and demonstrated the distinctive effect of these genes on mineral weathering by F77. IMPORTANCE Bacteria play important roles in mineral weathering and soil formation, although the molecular mechanisms underlying the interactions between bacteria and silicate minerals are poorly understood. In this study, the interactions between biotite and the highly effective mineral-weathering bacterium P. azotoformans F77 were characterized. Our results showed that the genes involved in gluconic acid metabolism and pilus formation play important roles in mineral weathering by F77. The presence of biotite could promote the expression of these genes in F77, and a distinctive effect of these genes on mineral weathering by F77 was observed in this study. Our results provide new knowledge and promote better understanding regarding the interaction between silicate minerals and mineral-weathering bacteria, as well as the molecular mechanisms involved in these processes.

The variations in flavor substances across the different stages of fermented soybean whey tofu (FSWT) production were analyzed by headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) combined with principal component analysis (PCA). The results revealed 24 representative flavor compounds in the samples across all production stages. After heating, the signal intensity of hexanal, 1-octen-3-ol, heptanal, and (E)-2-hexenol, which are unpleasant flavor substances found in raw soymilk, weakened, whereas those of some aroma substances increased. Furthermore, fermented flavor compounds, namely, 2-heptanone, 2-pentylfuran, pentanal, and 2,3-butanedione, were produced after the addition of fermented soybean whey as a coagulant. A PCA based on the signal intensity of the detected volatile compounds revealed effective differentiation of samples from different stages into comparatively independent spaces. These results showed that the flavor fingerprints of the samples from different stages of FSWT production can be successfully built using HS-GC-IMS and PCA based on the detected volatile compounds.

In recent years, some marine microbes have been used to degrade diesel oil. However, the exact mechanisms underlying the biodegradation are still poorly understood. In this study, a hypothermophilous marine strain, which can degrade diesel oil in cold seawater was isolated from Antarctic floe-ice and identified and named as Rhodococcus sp. LH. To clarify the biodegradation mechanisms, a gas chromatography-mass spectrometry (GC-MS)-based metabolomics strategy was performed to determine the diesel biodegradation process-associated intracellular biochemical changes in Rhodococcus sp. LH cells. With the aid of partial least squares-discriminant analysis (PLS-DA), 17 differential metabolites with variable importance in the projection (VIP) value greater than 1 were identified. Results indicated that the biodegradation of diesel oil by Rhodococcus sp. LH was affected by many different factors. Rhodococcus sp. LH could degrade diesel oil through terminal or sub-terminal oxidation reactions, and might also possess the ability to degrade aromatic hydrocarbons. In addition, some surfactants, especially fatty acids, which were secreted by Rhodococcus into medium could also assist the strain in dispersing and absorbing diesel oil. Lack of nitrogen in the seawater would lead to nitrogen starvation, thereby restraining the amino acid circulation in Rhodococcus sp. LH. Moreover, nitrogen starvation could also promote the conversation of relative excess carbon source to storage materials, such as 1-monolinoleoylglycerol. These results would provide a comprehensive understanding about the complex mechanisms of diesel oil biodegradation by Rhodococcus sp. LH at the systematic level.

The Angoumois grain moth, Sitotroga cerealella (Olivier, 1789) (Lepidoptera: Gelechiidae), is primarily a pest of stored products, that feeds inside the grain as larvae inducing significant economic loss in various stored commodities. Our previous studies proved that garlic essential oil and its active substances inhibit oviposition in moths. To further explore the effect on reproductive behavior and accurately interpret the results in terms of effective control of the moth population, moths were treated with diallyl trisulfide (DATS), an active substance of garlic essential oil, at a dose of 0.015 µL/L in air (LC20, sub-lethal concentration). The results showed that fecundity and the proportion of viable eggs significantly decreased when the moths were treated with LC20 DATS. Furthermore, female circadian mating rhythms and calling periodicity changed significantly after treatment. Compared with controls, the peak in mating occurred approximately 1 h earlier on the first day after DATS treatment, while it was significantly later on days 2 and 3. Moreover, mating frequency declined in presence of DATS compared with the controls. The percentage of females engaging in calling behavior decreased significantly with time, to less than 50%, 2 days after treatment, while a high calling percentage (>80%) was recorded for control moths on all 4 days. In addition, DATS had an inhibitory effect on the mating duration of S. cerealella. Moreover, a significant reduction was observed in the amount of sex pheromones extracted 8 h and 9 h after treatment. Our findings suggested that DATS has the potential to manipulate the moth population at LC20 and would be an efficient alternative to synthetic insecticides for the control of pests having low toxicity to non-target organisms and ecosystems.

Inhibition of bone regeneration by wear debris is the main cause of peri-prosthetic osteolysis. Here, we investigated the effect of icariin on cell proliferation, apoptosis, osteogenic differentiation and matrix mineralization of osteoblasts in an in vitro model of titanium (Ti) particle-induced osteolysis. In the present study, MC3T3-E1 cells were pretreated with 10-8 M icariin for 4 h and then incubated with Ti particles (0.1 mg/mL). The results showed that Ti particles inhibited cell proliferation and promoted cell apoptosis of MC3T3-E1 cells, whereas icariin pretreatment blocked the effect of Ti particles. In addition, we found that icariin stimulation alone increased ALP activity, accelerated matrix mineralization and upregulated the levels of bone morphogenetic protein 2 (BMP2), Runt-related transcription factor 2 (Runx2), osteocalcin (OCN) and miR-21-5p; whereas, Ti particles alone exerted the opposite effects. Icariin partly reversed the effect of Ti particles on cell differentiation and mineralization. Twenty hours after transfection with antagomiR-21-5p or antagomiR-NC, the cells were pretreated with icariin for 4 h and then incubated with Ti particles. Further studies showed that partial knockdown of miR-21-5p abolished the promotion effect of icariin on osteoblast differentiation and matrix mineralization in Ti particle-stimulated MC3T3-E1 cells. In conclusion, miR-21-5p may be a potential pro-osteogenesis regulator and icariin may protect against Ti particle-induced inhibition of osteogenic differentiation and mineralization through upregulation of miR-21-5p.

The angoumois grain moth, Sitotroga cerealella Olivier, is a serious stored-grain pest worldwide. In previous studies, we demonstrated that diallyl trisulfide (DATS), one of active substances from garlic essential oil, inhibited oviposition of S. cerealella, but the mechanism for this inhibition remains unclear. We suspected that DATS-regulated gene expression changes in S. cerealella may be a reason for the modulation of fertility and reproductive behaviors of the moth. Comparative transcriptomes of female S. cerealella were built by RNA-sequencing to detect differences between DATS exposure and background air with special lights on the key genes related to the fecundity. A total of 304 differentially expressed genes (DEGs) were detected in female moth after exposure to DATS, of which 120 were up-regulated, and 184 were down-regulated. Sixty-two of these DEGs encoding proteins involved in growth and development of eggs and regulation of hormones and mating behavior were identified, and of these, 49 were annotated as chorion protein. Meanwhile, quantitative real-time PCR verified that DEG data were reliable for further transcriptional analysis. Additionally, the results of ovarian dissection and oviposition activity bioassay suggest that DATS has negative effects on ovarian development and fecundity of S. cerealella. Our comprehensive sequence resource with desirable quality enriched the genomic platform of S. cerealella, and the DEGs that were identified will facilitate understanding of the molecular mechanism of ovipositional inhibition by DATS.

Ai-Yue Zhao,1 Yi-Jun J Dai,1 Jian-Feng Lian,2 Yan Huang,1 Jian-Guang Lin,1 Yang-Bin Dai,1 Tian-Wen Xu1 1Department of Medical Oncology, the Second Affiliated Hospital of Fujian Medical University, Quanzhou, China; 2Department of Anesthesiology, Quanzhou Children’s Hospital, Quanzhou 362000, China Background: Yes-associated protein (YAP), a key player of the Hippo pathway, has been identified to have more and more important roles in tumorigenesis and may be an important biomarker for cancer therapy. YAP is important for bladder cancer cell migration, metastasis, and drug resistance; however, its function in bladder cancer stem cells remains unknown. Purpose: The aim of this work was to examine the expression and role of YAP in bladder cancer stem cells. Materials and methods: We identified that the expression level of YAP was significantly enriched in bladder cancer stem cells compared to noncancer stem cell population. Moreover, the effect of YAP on stem cell self-renewal was examined in bladder cancer cells by siRNA silencing approach. In addition, we showed that YAP is required for aldehyde dehydrogenase activity in bladder cancer cells. Results: RNAseq analysis and quantitative real-time PCR results showed that silencing of YAP inhibited the expression of ALDH1A1 gene. Conclusion: Collectively, our findings for the first time elucidated that YAP serves as a cancer stem cell regulator in bladder cancer, which provided a promising therapy strategy for patients with bladder cancer. Keywords: bladder cancer, YAP, ALDH1, cancer stem cells

To assess the time-dependent stress evidence in dynamic allocation of physiological metabolism of Nilaparvata lugens nymphs in response to elevated CO2, we measured the time-dependent allocation of nutrient compositions and physiological metabolism in the bodies of N. lugens at 1h, 4h and 12h under elevated CO2. Elevated CO2 significantly increased the contents of nutrient compositions (protein, glucose and total amino acids) and catalase (CAT) enzyme activity in the body of N. lugens at 12h relative to 1h and 4h (P

Nuclear factor Y (NF-Y) is a ubiquitous transcription factor found in many eukaryotic organisms. NF-Y consists of three distinct subunits, namely, NF-YA, NF-YB, and NF-YC. In this study, a NF-YB gene, AsNF-YB3, containing the full-length coding region was isolated from garlic. A sequence alignment and phylogenetic analysis showed that AsNF-YB3 clustered with TaNF-YB2 and AtNF-YB3. AsNF-YB3 was expressed in various organs, including the roots, cloves and leaves. A real-time RT-PCR analysis revealed that GA3, ABA and MeJA treatment upregulated the expression of AsNF-YB3 transcript. Overexpressing AsNF-YB3 in tobacco plants significantly accelerated germination compared to the wild type. Moreover, the total soluble sugar, fructose and sucrose levels were significantly higher in transgenic tobacco lines that constitutively overexpressed AsNF-YB3, suggesting that AsNF-YB3 might be involved in carbohydrate metabolism. Transgenic tobacco plants that overexpressed AsNF-YB3 displayed enhanced root elongation and shoot growth. The overexpression of AsNF-YB3 also strongly promoted early flowering in tobacco. The results of this study in tobacco suggest that garlic AsNF-YB3 likely plays an important role as a plant growth regulator.

The angoumois grain moth, Sitotroga cerealella, is one of the most serious stored-grain pests worldwide. Control of this moth may be achieved by interfering with olfactory pathways to disrupt male-female communication with sex pheromones, using plant volatiles like garlic or its active substances. Here, three odorant-binding protein (OBP) genes [namely Si. cerealella general OBP 1 (ScerGOBP1), ScerGOBP2 and Si. cerealella pheromone-binding protein (ScerPBP)] were cloned from Si. cerealella antennae, and quantitative real-time PCR showed that these genes were predominantly expressed in adult antennae. ScerPBP expression was male-biased, but ScerGOBP1 and ScerGOBP2 were similar between sexes. The results of competitive binding assays indicated that a garlic substance, diallyl trisulphide (DATS), had similar or even higher binding affinity to ScerPBP than Si. cerealella sex pheromone, 7Z, 11E-hexadecadien-1-ol acetate (HDA). In olfactometer bioassays, DATS significantly reduced the response of adults to HDA when they were exposed to air filled with HDA and DATS. Surprisingly, ScerGOBP2, which is postulated to be involved in the detection of general odours, displayed higher affinity with HDA than did ScerPBP, indicating that ScerGOBP2 may also have a role in pheromone perception. These data suggest that DATS may interfere with recognition of female-produced sex pheromone, disrupting female and male mating behaviour and resulting in a new idea for controlling stored grain pests.

A hydroponic study was conducted to determine the effects of selenium (Se: 0, 3, 6 μmol L –1 ) on senescence-related oxidative stress in garlic plants grown under two sulfur (S) levels. We evaluated the yields of plants harvested at 160 and 200 days after sowing. Plants grown under a low Se dose (0.3 μmol L –1 ) at low S level showed higher yields (12.0% increase in fresh weight yield, 13.7% increase in dry weight yield) than the controls, despite a decrease in chlorophyll concentration. Compared with control plants, the Se-treated plants showed lower levels of lipid peroxidation. The Se-treated plants also showed higher activities of glutathione peroxidase and catalase, but lower superoxide dismutase activities. Changes in F v / F m values and proline contents were affected more strongly by S than by Se. On the basis of our results, we can conclude that Se plays a key role in the antioxidant systems in garlic seedlings. It delays senescence by alleviating the peroxide stress, but it can be toxic at high levels. A high S level may increase tolerance to high Se concentrations through reducing Se accumulation in plants.

During the batch bioethanol fermentation process, although Saccharomyces cerevisiae cells are challenged by accumulated ethanol, our previous work showed that the ethanol tolerance of S. cerevisiae increased as fermentation time increased. However, the exact molecular mechanisms underlying the increased ethanol tolerance of S. cerevisiae are still poorly understood. In this study, a gas chromatography-mass spectrometry-based metabolomics strategy was used to determine the fermentation process-associated intracellular metabolic changes in S. cerevisiae cells. With the aid of partial least squares-discriminant analysis between two of the three fermentation stages (i.e., the lag, exponential, and stationary phases), 40 differential metabolites with variable importance and a projection value greater than 1 were identified. During the bioethanol fermentation process, S. cerevisiae cells could continuously remodel their membrane composition and structure to obtain higher ethanol tolerance. During the lag–exponential phase transition, in spite of a down-regulated TCA cycle, the increased ergosterol content combined with decreased saturated fatty acid content might be the most significant factor in making yeast cells more robust and ethanol-tolerant. During the exponential–stationary phase transition, a re-activated TCA cycle could provide plenty of energy, and the increased energy production together with the increased energy requirements might be partly responsible for the increased ethanol tolerance in the stationary phase. Moreover, the increased content of glycerol, trehalose, ergosterol and some amino acids also might jointly confer the yeast cells with higher ethanol tolerance. These results highlighted our knowledge about the relationship between the bioethanol fermentation process and ethanol tolerance, and could contribute to the construction of feasible ethanologenic strains with higher ethanol tolerance.

This study investigated the effects of diallyl trisulfide (DAT), an active substance from garlic essential oil, on the metabolism of the main energy substances of pre- and postmating males of Sitotroga cerealella. Males at 12 h postemergence were fumigated with DAT at a concentration (LC10 = 0.010 µL/L) in a glass jar for 7 h. The main energy metabolites from pre- and postmating males were determined, including protein, triglyceride, glycogen, total soluble sugar, trehalose, and trehalase. The contents of total protein and total soluble sugar and the trehalase activity of premating males were significantly increased following DAT treatment, whereas the contents of protein from the accessory gland, triglyceride, glycogen, and trehalose were significantly decreased after treatment. Additionally, after mating, the total protein and soluble sugar contents were significantly increased and the glycogen content was significantly decreased in the treatment group relative to the levels in controls, but there was no significant difference observed in triglyceride, accessory gland proteins, trehalose content, or trehalase activity between the treatment and control groups. Furthermore, the changes in the main energy substances between pre- and postmating in males after the DAT treatment (∆DAT) were smaller than those in the control group (∆CK). This result indicated that DAT can accelerate the rate of metabolism in males at LC10, leading to the accumulation of greater levels of total soluble sugar to support life activities and to the increased synthesis of proteins to resist an adverse environment.

Summary The adult CNS has poor ability to replace degenerated neurons following injury or disease. Recently, direct reprogramming of astrocytes into induced neurons has been proposed as an innovative strategy toward CNS repair. As a cell population that shows high diversity on physiological properties and functions depending on their spatiotemporal distribution, however, whether the astrocyte heterogeneity affect neuronal reprogramming is not clear. Here, we show that astrocytes derived from cortex, cerebellum, and spinal cord exhibit biological heterogeneity and possess distinct susceptibility to transcription factor-induced neuronal reprogramming. The heterogeneous expression level of NOTCH1 signaling in the different CNS regions-derived astrocytes is shown to be responsible for the neuronal reprogramming diversity. Taken together, our findings demonstrate that region-restricted astrocytes reveal different intrinsic limitation of the response to neuronal reprogramming., Highlights • Region-restrict astrocytes (ACs) exhibit obvious heterogeneity • Region-restrict ACs show distinct susceptibility to neuronal reprogramming • AC heterogeneity does not affect the maturation of induced neurons • Notch1 is involved in the neuronal reprogramming diversity of region-restrict ACs, Su and colleagues show that astrocytes derived from cortex, cerebellum, and spinal cord exhibit biological heterogeneity and possess distinct susceptibility to transcription factor-induced neuronal reprogramming. The heterogeneous expression level of Notch1 signaling in the different CNS regions-derived astrocytes is shown to be responsible for the neuronal reprogramming diversity.

Species of the genus Garra from the Irrawaddy and Salween River basins in west Yunnan, China are difficult to identify and their taxonomy remains problematic. We conducted a systematic review of the genus using a combination of molecular and morphological approaches to address these issues and describe a new species, G. surgifrons . Phylogenetic analyses indicate that 15 species of Garra do not form a monophyletic group. Garra imberba and G. nujiangensis form a distinct clade that is not sister to the remaining species of Garra . According to the combination of morphological characters of the proboscis and rostrum, Garra from western Yunnan Province can be divided into 4 groups: (1) smooth snout; (2) unilobed proboscis; (3) bi-lobed proboscis; and (4) tri-lobed proboscis. The types of snout shapes in Garra are consistent with the topological structure of the phylogenetic tree. Garra surgifrons can be distinguished from all other congeners occurring in China and Southeast Asia by a combination of the following characters: presence of a transverse lobe and a tri-lobed proboscis; both dorsal and caudal fins without black markings; the proboscis short and broad, a roughly quadrate shape from the dorsal view with a poorly developed and slightly protuberant middle lobe, with the front tip not suspended from the lateral view and the width almost equal to the width of the lateral lobes; the posterior margin of mental adhesive disc extending close to or reaching the vertical through the central line of the eye and never reaching the posterior margin of the eye; two pairs of barbels; 16 circumpeduncular scales; and 32–34 lateral line scales.

Background Primary liver cancer (PLC) is the third largest contributor to cancer mortality in the world. PLC is a heterogeneous disease that encompasses several biologically distinct subtypes including hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (ICC) and combined hepatocellular-cholangiocarcinoma (CHC). CHC is a distinct, albeit rare, subtype of PLC and is comprised of cells with histopathological features of both HCC and ICC. Several studies have focused on the mutation and expression landscapes of HCC and ICC. However, studies of CHC were rare. Objective The aim of the current study was to identify genetic and gene expression alterations in the carcinogenesis and development of CHC and ICC in the Chinese population. Unraveling both similar and differing patterns among these subtypes may help to identify personalized medicine approaches that could improve patient survival. Methods Whole genome sequencing (WGS), whole exome sequencing (WES) and RNA-seq were performed on 10 ICC and 10 CHC samples, matched with adjacent non-tumor liver tissue specimens. Comparative analysis was performed using HCC datasets from The Cancer Genome Atlas (TCGA). Results Mutational and transcriptional landscapes of CHC and ICC were clearly delineated. TP53 and CTNNB1 were identified as exhibiting mutations in CHC. ARID1A , PBRM1 , and IDH1 were frequently mutated in ICC. RYR3 , FBN2 , and KCNN3 are associated with cell migration and metastasis and might be driver genes in CHC. KCNN3 was identified as also exhibiting mutations in ICC. The ECM-receptor interaction pathway associated fibrogenic hepatic progenitor cell differentiation and liver fibrosis may play an important role in carcinogenesis of PLC. Chromatin remodeling and chromosome organization are key processes in carcinogenesis and development in PLC. P53 related pathways showed alterations in CHC and HCC. Inflammation may be a key factor involved in ICC carcinogenesis. Conclusion CHC and ICC are different subtypes of PLC. This study discusses predominantly the molecular genetic details of PLC subtypes and highlights the need for an accurate diagnosis and treatment of specific PLC subtypes to optimize patient management.

The aim of the present study was to investigate the expression of nesprin-1 protein in MSCs and its effects on the differentiation of rat bone-marrow mesenchymal stem cells (MSCs). Surface-associated antigens of MSCs were detected by flow cytometry. MSC differentiation was induced by treatment with 10 μmol/l 5-azacytidine. Sprague-Dawley rats were anesthetized prior to thoracotomy and subsequent ligation of the left anterior descending coronary artery to establish a model of myocardial infarction. Two weeks following myocardial infarction, DAPI-marked MSCs were injected into the infarcted region in the experimental group, while DMEM was injected into the infarcted region of the control group. Characteristics of the putative cardiac-myogenic cells were evaluated using immunohistochemical and immunofluorescent analysis. The messenger RNA expression levels of cardiac-myogenic specific genes; desmin, α-actinin and cardiac troponin I (cTnI) were detected by reverse transcription quantitative polymerase chain reaction. The expression of nesprin-1 protein in MSCs was identified by immunofluorescence and western blot analysis, prior to and following MSC differentiation. Following differentiation, the MSCs appeared spindle-shaped with irregular processes and were positive for CD90 and CD29, but negative for CD45. Cardiomyocyte-like cells were positive for desmin, α-sarcomeric actin and cTnI. Nesprin protein was detected in the nuclear membrane via immunofluorescence, and following MSC differentiation into cardiomyocyte-like cells, the expression of nesprin protein was significantly higher (*P=0.03

The adult mammalian CNS has a limited capacity to regenerate after traumatic injury. In this study, a combinatorial strategy to promote axonal regeneration and functional recovery after spinal cord injury (SCI) was evaluated in adult rats. The rats were subjected to a complete transection in the thoracic spinal cord, and multichannel scaffolds seeded with activated Schwann cells (ASCs) and/or rat bone marrow-derived mesenchymal stem cells (MSCs) were acutely grafted into the 3-mm-wide transection gap. At 4 weeks post-transplantation and thereafter, the rats receiving scaffolds seeded with ASCs and MSCs exhibited significant recovery of nerve function as shown by the Basso, Beattie and Bresnahan (BBB) score and electrophysiological test results. Immunohistochemical analyses at 4 and 8 weeks after transplantation revealed that the implanted MSCs at the lesion/graft site survived and differentiated into neuron-like cells and co-localized with host neurons. Robust bundles of regenerated fibers were identified in the lesion/graft site in the ASC and MSC co-transplantation rats, and neurofilament 200 (NF) staining confirmed that these fibers were axons. Furthermore, myelin basic protein (MBP)-positive myelin sheaths were also identified at the lesion/graft site and confirmed via electron microscopy. In addition to expressing mature neuronal markers, sparse MSC-derived neuron-like cells expressed choline acetyltransferase (ChAT) at the injury site of the ASC and MSC co-transplantation rats. These findings suggest that co-transplantation of ASCs and MSCs in a multichannel polymer scaffold may represent a novel combinatorial strategy for the treatment of spinal cord injury.

The Angoumois grain moth, Sitotroga cerealella, is a serious pest of stored grains worldwide. Presently, the best effective control against the moth is to disrupt the sexual communication between sexes. Sexual communication in moths includes two processes in which females produce and release pheromones from the sex pheromone gland and males detect and respond to them with a relatively sophisticated olfactory system in their antennae. To better understand these processes, we studied the ultrastructure of antennal and ovipositor sensilla of S. cerealella and determined the location of the female sex pheromone gland. Seven types of antennal sensilla were identified on both sexes: sensilla trichodea, sensilla chaetica, sensilla coeloconica, sensilla styloconica, sensilla auricillica, sensilla squamiformia and Bӧhm bristles. Of these sensilla, the sensilla trichodea were significantly more abundant on male antennae than on those of females, suggesting that these sensilla may detect the sex pheromones. On the ovipositor, only sensilla chaetica of various lengths were found. The sexual gland was an eversible sac of glandular epithelium that was situated dorsally in the intersegmental membrane between the 8th and 9th abdominal segments. These results will lead to a better understanding of mate finding with sex pheromones for this worldwide pest species.

The aim of this study was to investigate the expression of nesprin‑1 protein and its effects on rat bone marrow mesenchymal stem cells (MSCs). MSCs were cultured in DMEM and surface-associated antigens of MSCs were detected by flow cytometry. The protein expression of nesprin-1 was detected by immunofluorescence and western blot analysis. A lentiviral vector expressing small interfering RNA (siRNA) targeting nesprin-1 was constructed (LV-siNesprin-1) and the MSCs were subsequently transfected with this vector. Another group of MSCs was transfected with the LV-GFP vector and another group of untransfected cells was used as the controls (normal group). The protein expression level of nesprin-1 in the 3 groups of MSCs (LV-siNesprin-1, LV-GFP and normal group) was measured by western blot analysis. Cell proliferation was assessed by MTT assay, and the cell cycle and apoptosis were detected by flow cytometry. DAPI was used to stain the nucleus of the MSCs. The MSCs appeared spindle-shaped with irregular processes and were positive for CD90, CD29 and negative for CD45. Nesprin-1 protein was found in the nuclear membrane. The protein expression of nesprin-1 in the LV-siNesprin-1 group was lower than that in the LV-GFP (P=0.03) and normal group (P=0.028); this difference was significant (P

The effects of aerobic and anaerobic conditions on polyhydroxybutyrate (PHB) and glycerin yields during the course of lignin degradation were studied with Pseudomonas stutzeri P156. The lignin degradation rate was 22.5% and 15%, respectively, under aerobic and anaerobic condition. However, the gradation rate/biomass (YL/X) was nearly no differences that diversified from 0.9% to 3%. In addition, the maximal anaerobic yield of PHB and glycerin yield was 0.42 g/g and 56.3 ug/ml, which was 176.6% and 36.7% higher than that under aerobic condition, respectively. Interestingly to point out, anaerobic lignin degradation rate was decreased by 33.3% relative to the aerobic one, but lignin content did decreased by 15% at the cost of higher yields of PHB and glycerin, which suggested biosyntheses of PHB and glycerin could balance the redox potential under anaerobic condition, facilitate lignin degradation by Pseudomonas stutzeri P156, persistently, and meet the process demands of straw-based anaerobic methane fermentation.

To investigate the relationship between nuclear factor Y (NF-Y) and stress tolerance in garlic, we cloned a NF-Y family gene AsNF-YC8 from garlic, which was largely upregulated at dehydrate stage. Expression pattern analyses in garlic revealed that AsNF-YC8 is induced through abscisic acid (ABA) and abiotic stresses, such as NaCl and PEG. Compared with wild-type plants, the overexpressing-AsNF-YC8 transgenic tobacco plants showed higher seed germination rates, longer root length and better plant growth under salt and drought stresses. Under drought stress, the transgenic plants maintained higher relative water content (RWC), net photosynthesis, lower levels of malondialdehyde (MDA), and less ion leakage (IL) than wild-type control plants. These results indicate the high tolerance of the transgenic plants to drought stress compared to the WT. The transgenic tobacco lines accumulated less reactive oxygen species (ROS) and exhibited higher antioxidative enzyme activities compared with wild-type (WT) plants under drought stress, which suggested that the overexpression of AsNF-YC8 improves the antioxidant defense system by regulating the activities of these antioxidant enzymes, which in turn protect transgenic lines against drought stress. These results suggest that AsNF-YC8 plays an important role in tolerance to drought and salt stresses.

Background/Aims: Adipose-derived stem cells (ASCs) belong to mesenchymal stem cells and may play a potential role as seeding cells in stem cell transplantation. To be able to exploit stem cells as therapeutic tool, their defects in some important cellular functions, such as low survival rate and cellular activity, should be considered. This is especially the case for stem cells that are intended for transplantation. Of note, stem cell responses to hormones should be considered since estrogen is known to play a critical role in stem cell behavior. However, different impacts of the estrogen receptor (ER) types α and β have not been fully determined in ASC function. In this study, we investigated effects of ERα and ERβ on ASC proliferation, migration, as well as in adipogenesis. Methods: ASCs obtained from mice were cultured with 100nM ERα or ERβ agonist PPT and DPN, respectively. The ERα and ERβ antagonist ICI 182,780 (100nM) was used as control. Results: Compared to ERβ, ERα appears more potent in improving ASC proliferation and migration. Investigation of adipogenesis revealed that ERβ played a significant role in suppressing ASC-mediated brown tissue adipogenesis which is in contrast to ERα. These results correlated with reduced mRNA expression of UCP-1, PGC-1α and PPAR-γ. Conclusions: ERα plays a more critical role in promoting ASC proliferation and migration while ERβ is more potent in suppressing ASC brown adipose tissue differentiation mediated by decreased UCP-1, PGC-1α and PPAR-γ expression.

Astronautical mutagenesis is a new and security agricultural breeding technique in recent decades. There are very large differences in physical environment between space and ground. After returning to the ground, new varieties are bred by the conventional breeding techniques. There are rich genetic types in mutagenesis. In this paper, “Longmu 801” alfalfa seeds were carried by “Shijian-8” recoverable satellite and some new types were selected and bred. The genetic variation of 11 mutated lines and their CK were identified by RAPD analysis, determination of leaf protein concentrates (LPC) and crude protein. For genetic identification, 100 primers were tested and 19 primers were selected in RAPD amplification. The amplified bands and polymorphic sites were rich. Polymorphism points and ratios of polymorphism point were statistical in detail. They were 326 and 15.04% respectively. For quality identification, it was compared with yields of alfalfa LPC and crude protein contents. The results showed that the range of yields of LPC was 6.52%-7.65%, range of nitrogen content was 43.79%-52.38%. Comparing with CK, variation ranges were -3.99%-+12.99% and -4.45%-+12.49%.

NK1.1+ natural killer (NK)-T cells reactive to CD1 appear to be involved in spontaneous autoimmune disease, but their role in induced autoimmune disease remains unclear. While we previously reported a regulatory role of NK cells in experimental autoimmune encephalomyelitis (EAE), we demonstrate here that NK-T cells would also play a pivotal role in the control of EAE. C57BL/6 (B6) mice selectively depleted for NK-T cells were generated by antibody-depended protocols or knocking out TCRJα281 gene. Regardless of the method for NK-T cell deletion, these NK-T depleted mice developed unusually early onset of EAE (5–8 days after challenge) after immunization with an encephalitogenic peptide myelin oligodendrocyte glycoprotein (MOG) 35–55 . The early EAE onset was associated with early induction of T helper cell type 1 (Th1) cells and marked elevation of Th1 cytokines in the serum. It was of note that, although the NK-T deficient mice spontaneously recovered from EAE, mice selectively depleted for NK cells developed a later onset of non-remitting EAE. These data establish differential roles played by NK-T and NK cells in the protection against EAE.

The angoumois grain moth, Sitotroga cerealella (Olivier), is one of the most serious stored grain pests around the world. In attempts to reduce the losses caused by the moth and to suppress its populations, the fumigant activities, behavioral influence and ovipositional inhibition of garlic (Allium sativum) essential oil and its two major components, diallyl disulfide and diallyl trisulfide, were investigated against the adult grain moth. Their effects on reduction in survival of first instar larvae to adult emergence were also evaluated. Results showed that these three materials (garlic essential oil, diallyl disulfide and diallyl trisulfide) had significant fumigant activity with 50% lethal concentration values at 1.33, 0.99, and 1.02 μL/L air space, respectively; meanwhile, the three materials possessed high behavioral deterrent activities against adults in the Y-tube olfactometer. When applied to rice grains, these materials reduced adult longevity and inhibited oviposition, with ovipositional inhibition above 70% at a concentration of 1.5 μL/25 g in either no-choice or two-choice tests. In short, the study showed that both diallyl disulfide and diallyl trisulfide, like garlic essential oil, acted as fumigants, produced behavioral deterrence and inhibited oviposition against angoumois grain moth. Our work here indicates that diallyl disulfide and diallyl trisulfide may serve as potential alternatives for grain protectants since both of them can be prepared easily from readily available chemicals.

Laboratory bioassays were carried out to determine the efficacy of garlic, Allium sativum L. (Amaryllidaceae), essential oil applied alone or with diatomaceous earth (DE) against adult rice weevils, Sitophilus oryzae (L.) (Coleoptera: Curculionidae) and red flour beetles, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). The results showed that the combination treatment was significantly more effective than either treatment alone. In addition, the results also showed that the simultaneous application of essential oil plus DE significantly reduced the concentration of essential oil alone required for an effective treatment and the application rate of DE can be reduced when combined with essential oil. Moreover, the activity of the combination treatment lasted longer than that of essential oil alone and the survival of eggs or larvae to adult stage was significantly inhibited in the combined treatments against both species, compared with the use of essential oil alone. Our results suggested that garlic essential oil combined with DE has a strong additive effect, and therefore may have potential as an alternative to synthetic insecticides for the control of insect pests of stored products.

In this study, the fumigant activity of garlic essential oil and its major components, diallyl disulfide, diallyl trisulfide, and diallyl sulfide, against the stored-product insect, Tribolium castaneum, were investigated. Results from the space fumigation revealed that garlic essential oil and diallyl trisulfide had strong fumigant activities against the adult insect. In glass chambers (10 cm in diameter × 125 cm in height) filled with 10, 20, 30, 40, and 50% volume of rough rice, the fumigant activity of diallyl trisulfide was more potent than that of garlic essential oil. At the concentration of 8 μl/l and 50% filling ratio, to obtain 100% mortality, exposure time of only 4 days was needed for diallyl trisulfide, but a longer exposure time (6–7 days) was required for garlic essential oil. Meanwhile, the fumigant effects on the germination rate of kernels exposure were ascertained. Results proved that there were no side effects for these substances on germination rate of kernels exposed 6 months after fumigation. In addition, the fumigant effect on F1 progeny production in rough rice was also evaluated for garlic essential oil and diallyl trisulfide. The results showed that progeny production was totally suppressed at 4 μl/l of these two substances, either at 10 or 50% filling ratio. The results may provide valuable advances for future fumigant development and possible utility as a fumigant especially as a seed protectant in warehouses filled with grain.

Over the past few decades genetic engineering has been applied to improve cotton breeding. Agrobacterium medicated transformation is nowadays widely used as an efficient approach to introduce exogenous genes into cotton for genetically modified organisms. However, it still needs to be improved for better transformation efficiency and higher embryogenic callus induction ratios. To research further the difference of mechanisms for morphogenesis between embryogenic callus and non-embryogenic callus, we carried out a systematical study on the histological and cellular ultrastructure of Agrobacterium transformed calli. Results showed that the embryogenic callus developed nodule-like structures, which were formed by small, tightly packed, hemispherical cells. The surface of some embryogenic callus was covered with a fibrilar-like structure named extracellular matrix. The cells of embryogenic calli had similar morphological characteristics. Organelles of embryogenic callus cells were located near the nucleus, and chloroplasts degraded to proplastid-like structures with some starch grains. In contrast, the non-embryogenic calli were covered by oval or sphere cells or small clusters of cells. It was observed that cells had vacuolation of cytoplasm and plastids with a well organized endomembrane system. This study aims to understand the mechanisms of embryogenic callus morphogenesis and to improve the efficiency of cotton transformation in future.

The interaction of baicalin and bovine serum albumin (BSA) was investigated using fluorescence spectroscopy (FS), resonance light scattering spectroscopy (RLS), and ultraviolet spectroscopy (UV). The apparent binding constants (Ka) between baicalin and BSA were 1.67 × 106 (22 °C), 1.98 × 106 (32 °C) and 2.01 × 106 (42 °C), and the binding sites values (n) were 1.33 ± 0.01. According to the Forster theory of nonradiation energy transfer, the binding distances (r) between baicalin and BSA were 1.94, 1.95 and 1.96 nm at 22, 32, and 42 °C, respectively. The experimental results showed that the baicalin could be inserted into the BSA, quenching the inner fluorescence by forming the baicalin–BSA complex. The addition of increasing baicalin to BSA solution leads to the gradual enhancement in RLS intensity, exhibiting the formation of the aggregate in solution. It was found that both static quenching and non-radiation energy transfer were the main reasons for the fluorescence quenching. The entropy change and enthalpy change were positive, which indicated that the interaction of baicalin and BSA was driven mainly by hydrophobic forces. The process of binding was a spontaneous process in which Gibbs free energy change was negative.

A rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) using resonance light scattering (RLS) on an ordinary fluorescence spectrometer was developed. The viable MRSA reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to produce insoluble particles which displayed intense resonance scattering light. It showed a linear relationship between the number of viable MRSA and the RLS intensity. Dead MRSA were unable to reduce MTT. MRSA exposed to flavonoids extracted from Marchantia convoluta (MCF) showed a MCF concentration-dependent inhibition of the ability to reduce MTT. The RLS could, in combination with the MTT assay, be a rapid and sensitive detection method for vitro-cultured MRSA.

HeLa cell line, which was derived from cervical carcinoma, provides an idea platform to study both the integration of human papillomavirus and the massive mutations occurring on the cancer cell genome. Proteogenomics is a field with the intersection of proteomics and genomics to perform gene annotation and identify gene mutation. In this work, we first identified the SNV/INDEL, structural variation (SV), and virus infection/integration events from RNA-Seq data of HeLa cell line; then, by applying proteogenomics strategy, we were able to detect some of the genomic events with the tandem mass spectrometry (MS/MS) data from the same sample. Furthermore, some of the mutated peptides were experimentally validated using multiple reaction monitoring technology. The integrated analysis of the RNA-Seq and MS/MS data not only renders the discovery of HeLa cell genome variations more credible but also illustrates a practical workflow for protein-coding mutation discovery in cancer-related studies.

The aim of present study was to define the effects of insulin on atrial dynamics and ANP release and its modification in diabetic rats. An isolated perfused beating atrial model was used from control and diabetic rats. Insulin was perfused with and without an inhibitor for tyrosine kinase or phosphatidylinositol 3-kinase (PI 3-kinase). Insulin increased the release of ANP and decreased atrial contractility in a dose-dependent manner. During the perfusion of 10−10 M insulin, the release of ANP abruptly increased within 8 min by approximately 40% and then decreased with time despite of continuous perfusion. In terms of increasing the dose of insulin, the time to reach the peak effect became faster and the slope to decrease became slower. In contrast, atrial contractility was gradually decreased with time. These effects were independent upon extracellular glucose. Genistein (10−5 M) or lavendustin C (10−5 M), a tyrosine kinase inhibitor, attenuated the release of ANP stimulated by insulin (10−8 M). Wortmannin (10−7 M) or LY294002 (10−5 M), a PI 3-kinase inhibitor, also attenuated insulin-stimulated ANP release. However, both inhibitors for PI 3-kinase and tyrosine kinase did not cause any significant effects on negative inotropism by insulin. Insulin-stimulated ANP release was augmented in streptozotocin-treated rat atria. The density of insulin receptor markedly increased in diabetic hearts. These results suggest that insulin stimulates the release of ANP through PI 3-kinase and tyrosine kinase, and augmentation of insulin-stimulated ANP release in diabetic rat atria may be partly due to an upregulation of insulin receptor.

Objective: To observe the effects of arsenic trioxide (As2O3) on human renal cell carcinoma (RCC) lines in vitro and to explore its possible molecular mechanisms.Methods: The microculture tetrazolium (MTT) assay was used to determine the anti-proliterative effects of As2O3 on human RCC lines. Flow cytometry was performed to investigate the effects of As2O3 on cell cycle and cell apoptosis. The reverse transcription-polymerase chain reaction (RT-PCR) was conducted to detect mRNA expression of Bcl-2, Bax, p53 and c-myc.Results: As2O3 inhibited the growth of RCC lines in vitro in a concentration-dependent manner. At the concentrations of 0.5, 1.0, 2.0 and 4. 0 μmol/L, the inhibition rates of As2O3 on RCC-WCS cells were 27.60%, 30.09%, 41.03% and 50.77%, respectively. Compared with untreated RCC-WCS, there was significant difference at each concentration (P

Previous studies suggest that osteopontin (OPN) levels are associated with the incidence and development of multiple types of cancer. This study analyzes the correlation between OPN gene variants and nasopharyngeal carcinoma (NPC). A total of 150 NPC patients and 150 healthy adults were enrolled and divided into NPC and control groups. In both groups, four OPN polymorphisms including rs1126772, rs4754, rs11728697 and rs9138 were quantitatively genotyped by SNaPshot SNP genotyping assay. Allele frequencies of four OPN polymorphisms were compared between two groups. No statistical significance was noted regarding genotype frequency of four OPN polymorphisms between male NPC and healthy subjects (all P > 0.05). Female subjects between two groups significantly differed in terms of genotype frequency of rs1172869 (P = 0.002). No statistical significance was observed in allele frequencies in each of four OPN polymorphisms between two groups (all P > 0.05). For NPC patients, the polymorphisms information content (PIC) ranged from 0.2961 to 0.3584 and 0.3047 to 0.3655 for normal controls. Additionally, rs11728697 had the highest effective number of alleles (Ne), expected heterozygosity (HE) and PIC in two groups. Female subjects in two groups significantly differed in the genotype frequency of rs1172869 while no statistical significance was noted for male counterparts. All subjects in the NPC and control groups had similar allele frequency. All the four OPN polymorphisms were moderately polymorphic. The mean NA, Ne, Ho, He and PIC of each OPN polymorphism in the control group were higher than those in the NPC group without a significant difference.

This report is designed to explore the roles of caspase-8, -9 and -3 in artemisinin (ARTE)-induced apoptosis in non-small cell lung cancer cells (A549 cells). ARTE induced reactive oxygen species (ROS)-mediated apoptosis in dose- and time-dependent fashion. Although ARTE treatment did not induce Bid cleavage and significant loss of mitochondrial membrane potential, it induced release of Smac and AIF but not cytochrome c from mitochondria, and silencing of Bak but not Bax significantly prevented ARTE-induced cytotoxicity. Moreover, ARTE treatment induced ROS-dependent activation of caspase-9, -8 and -3. Of the utmost importance, silencing or inhibiting any one of caspase-8, -9 and -3 almost completely prevented ARTE-induced activation of all the three caspases and remarkably abrogated the cytotoxicity of ARTE, suggesting that ARTE triggered an amplification activation loop among caspase-9, -8 and -3. Collectively, our data demonstrate that ARTE induces a ROS-mediated amplification activation loop among caspase-9, -8 and -3 to dominantly mediate the apoptosis of A549 cells.