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A novel approach was developed for the synthesis of 5H-isochromeno [4,3-b]indolizin-5-ones (ICIDOs) 3 through a cascade reaction of 1,2,3-indantriones 1 with ethyl 2-(pyridine-2-yl)acetates 2. Various 5H-isochromeno [4,3-b]indolizin-5-ones were produced by heating of the mixture of 1,2,3-indantriones and ethyl 2-(pyridine-2-yl)acetates for approximately 8 ​h in an environmentally-friendly PEG200 solvent with catalyst of a few drops of acetic acid. This approach has several advantages such as the use of an environmentally-friendly solvent, simple and practical operation (with filtration and washing and no column chromatography separation), good yields (up to 91%), use of commercially available raw materials, diverse target compounds, and products with potential biological activity.

In this study, a B4C/6061Al neutron absorber composite (NAC) tube containing 5 wt.% B4C particle was first fabricated by spark plasma sintering (SPS) followed by hot spinning (HS), then its microstructure and mechanical properties were experimentally investigated. It was found that, through spinning, B4C particles were better distributed in the 6061 Al matrix and the bonding of the B4C/6061Al matrix interface was improved. Dislocation pileups around B4C particles and dislocation loops were both observed. B4C particles could promote dynamic recrystallized nucleation and pin grain boundaries, resulting in grain refinement in the material. The yield strength (YS), ultimate tensile strength (UTS), and elongation of the spinned composite tube were found higher than that of the SPSed composite tube. The strength improvement of the fabricated B4C/6061Al neutron absorber composite tube was mainly due to the dislocation strengthening mechanism and grain refinement through spinning.

Meiotic crossover (CO) recombination between homologous chromosomes regulates chromosome segregation and promotes genetic diversity. Human females have different CO patterns than males, and some of these features contribute to the high frequency of chromosome segregation errors. In this study, we show that CO covariation is transmitted to progenies without detectable selection in both human males and females. Further investigations show that chromosome pairs with longer axes tend to have stronger axis length covariation and a stronger correlation between axis length and CO number, and the consequence of these two effects would be the stronger CO covariation as observed in females. These findings reveal a previously unsuspected feature for chromosome organization: long chromosome axes are more coordinately regulated than short ones. Additionally, the stronger CO covariation may work with human female-specific CO maturation inefficiency to confer female germlines the ability to adapt to changing environments on evolution.

Bigelovin (BigV) is a sesquiterpene lactone, isolated from Inula helianthus aquatica, which has been reported to induce apoptosis and show anti-inflammatory and anti-angiogenic activities. Nevertheless, the effects of BigV on liver cancer and the underlying mechanisms have not been investigated. In the study, we found that BigV exhibited potential anti-tumor activities against human liver cancer in vitro and in vivo. BigV reduced the cell proliferation and colony formation. BigV induced apoptosis through improving the cleavage of Caspase-3 and poly (ADP-ribose) polymerase 1 (PARP-1). The process was along with the activation of autophagy, as proved by the enhanced accumulation of autophagosomes, the microtubule-associated light chain 3B-II (LC3B-II) and Beclin-1, and p62 decrease. Further, the autophagy blockage markedly sensitized BigV-induced cell death, indicating the cytoprotective function of autophagy in liver cancer cell lines. In addition, BigV treatment inactivated the pathway of protein kinase B (AKT)/mammalian target of rapamycin (mTOR)/ribosomal protein S6 kinase (p70S6K). Of note, BigV-induced cell death was abolished by over-expressing the phosphorylation of mTOR. Intriguingly, the induction of apoptosis and autophagy were eliminated by the pretreatment of reactive oxygen species (ROS) scavenger N-acetyl- l -cysteine (NAC), suggesting that ROS played an important role in the regulation of BigV-induced cell death. Finally, in vivo studies demonstrated that BigV significantly suppressed the growth of HepG2 cancer xenograft tumors through the activation of apoptosis and autophagy in a dose-dependent manner with low systemic toxicity. In conclusion, the results revealed that BigV had significant antitumor effects against human liver cancer and it may potentially be used as a novel antitumor agent for the prevention of liver cancer.

Coordination polymers (CPs) are peculiar category of materials having multidimensional framework and could be used as selective and sensitive sensors for ions and molecules. In this regard, a new Zn(II)-based coordination polymer (CP) [Zn(H2L)(H2O)2]n (1) has been derived from 1,3-bis(3,5-dicarboxylphenoxy)benzene (H4L) ligand and spectroscopically characterized alongwith single crystal X-ray diffraction. The crystallographic experiments reveal the existence of 2D layer in 1. The CP retains its framework integrity in water and exhibits luminescent property. This newly designed CP has been used as luminescent sensor to detect antibiotics by the decline in its luminescence intensity. The sensing experiments indicated that the CP displays sensitive and selective sensing against nitrofurazone (NZF) with limit of detection (LOD) of 1.12 ppm and Ksv value of 2.41 × 103 M−1. The experimentally observed decline in the luminescence intensity of 1 in presence of antibiotics has been explained with the help of theoretical calculations.

Two new Ag(I) based coordination polymers (CPs), with formula [Ag2(H2L)(4,4′-bipy)2·2H2O] (1), and [Ag2(H2L)(bpz)2] (2) (H4L = 1,3-bis(3,5-dicarboxylphenoxy)benzene, 4,4′-bipy = 4,4′-bipyridine, and bpz = 3,3′,5,5′-tetramethyl-4,4′-bipyrazole) have been solvothermally synthesized and characterized. The single crystal X-ray structural analysis reveals that CPs 1 and 2 consist of 3D hydrogen-bonded organic framework with rectangle and grid channels, respectively. In addition, the photocatalytic properties of both 1 and 2 have been assessed under UV light against methyl orange (MO), rhodamine B (RB), methyl blue (MB), neutral red (NR), methylene blue (MB) are investigated in aqueous solutions. The results suggested that percentages of photocatalytic degradation toward MB, Rh B and MO in presence of 1 are 90.24%, 93.56% and 45.11%, respectively in 100 min. While in presence of 2 the percentage photodegradation of MB, Rh B and MO are 84.33%, 55.54% and 4.27% in 100 min. The plausible mechanistic pathways through which newly synthesized CPs executed photodegradation of these dyes have been explained with the aid of band gap calculations implementing density of states and partial density of states.

This study aims to investigate the effects of compositional gradient on microstructure and mechanical properties of a B 4 C/6061Al laminar composite fabricated using a powder metallurgy method - spark plasma sintering followed by extrusion and hot rolling. Results show that B 4 C particles were distributed fairly uniformly in the fabricated composites. Interfaces among different layers in the composites were bonded well and no porosity were observed in transition regions among different layers. The grains around B 4 C particles are smaller than those away from the B 4 C particles. Stress concentration is found to occur easily at the tip of the B 4 C particle, which facilitates the development of recrystallization nuclei. Compared with those of an extruded B 4 C/6061Al laminar composite, yield strength and ultimate tensile strength of the rolled B 4 C/6061Al laminar composite are obviously enhanced but its elongation to fracture is weakened. Two yielding points were observed in the rolled three-layer B 4 C/6061Al laminar composite due to the existence of B 4 C content gradient in the composite. Strengthening mechanisms in the fabricated laminar composites include grain refinement, dislocation strengthening, load transfer effect, and Orowan strengthening.

Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) has become an indispensable tool in analysis of macromolecule and small molecule in solid and liquid samples. However, there were few studies focusing on the ionization mechanisms especially for the small molecules. In this work, the compounds of pyrene, coronene and rubrene with similar molecular structures were used to investigate the ionization mechanism via SALDI-MS. Additionally, the effect of laser power on product ions was also investigated. It was found that strong cluster ions peaks nM+ (M = pyrene, coronene) were observed for pyrene and coronene, and daughter ions of coronene by loss of C2H2 were also observed. On the contrary, no cluster ion was obtained for rubrene, only daughter ions with the losses of nC6H5 (n = 1, 2, 3) were acquired. Finally, the ionization mechanism was discussed. The formation of clusters of pyrene and coronene was attributed to the interaction of π-π bonds. For rubrene, the spatial barrier weakened the interaction of π-π bonds because the four phenyl groups were not on the same plane of skeleton structure, thus impeding the formation of cluster ions.

Two pairs of rare benzofuran glucoside epimers, indicuses A and B and indicuses C and D, three biogenetically related compounds indicuses E–G, and one coumarin indicus H, as well as 11 known compounds, were isolated from the bark of Streblus indicus (Bur.) Corner. The structures of indicuses A–H were elucidated by NMR and MS data, as well as by CD. ( S )-Marmesinin exhibited moderate antimicrobial activity in vitro against Bacillus subtilis and Saccharomyces cerevisiae . 7,8-Dihydroxy-3-(3-methyl-2-butenyl) coumarin, umbelliferone, and scopoletin displayed strong cytotoxic activity in vitro against human bladder carcinoma cell line EJ. The structure-activity relationships indicate that hydroxylation at C-7 in the cytotoxic compounds is crucial to their activities.

Four new lignans, (7′ R ,8′ S )-4,4'-Dimethoxy-strebluslignanol ( 1 ), 3'-Hydroxy-isostrebluslignaldehyde ( 2 ), 3,3'-Methylene-bis(4-hydroxybenzaldehyde) ( 3 ), and 4-Methoxy-isomagnaldehyde ( 4 ), and six known lignans ( 5 – 10) , were isolated from the roots of Streblus asper . The structures of these molecules were elucidated through various spectroscopic methods of analysis, including 1D and 2D NMR. The stereochemistry at the chiral centres was determined using the CD spectrum and from coupling constant and optical rotation data. Compounds 1–6 showed good antimicrobial activity against Saccharomyces cerevisiae (ATCC 9763), Bacillus subtilis (ATCC 6633), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 11775), and Staphylococcus aureus (ATCC 25923), with MIC values ranging from 0.0150 to 0.0940 μM.

ANXA1 and EphA2 play a crucial role in cancer. Here, we report that ANXA1 competes with Cbl for binding EphA2 and increases its stability by inhibiting Cbl-mediated EphA2 ubiquitination degradation in nasopharyngeal carcinoma (NPC), leading to the enhanced NPC growth and metastasis in vitro and in vivo . In human NPC, patients with the high expression of both ANXA1 and EphA2 proteins have poorer disease-free survival and overall survival relative to patients with the high expression of one protein alone. Furthermore, based on the N-terminal amino acid 20-30 region of ANXA1 responsible for binding EphA2, we develop an ANXA1-derived peptide, named as A1 (20-30), that cuts its connection with EphA2, successfully downregulates EphA2 and suppresses NPC oncogenicity in vitro and in vivo. These findings reveal that ANXA1, via binding and stabilizing EphA2, drives NPC growth and metastasis, and present a strategy for targeting EphA2 degradation and treating NPC with a peptide.

Background: Gene-targeting therapy provides a novel therapeutic approach for tumor treatment using genetically-modified endothelial progenitor cells (EPCs) as cellular carriers. To study the therapeutic effect of EPCs armed with cytosine deaminase (CD) and endostatin (ES) fusion gene in liver cancer. Methods: EPCs derived from heart blood of mice were cultured and transfected with CD and ES fusion gene. EPCs, CD/ES, and CD/ES-EPCs were injected through their tail veins into mice with hepatoma, respectively. Subsequently, the in vivo tumor volumes were observed by MRI. Findings: Tumor volumes in the group injected CD/ES-EPCs were found to be smaller than other groups receiving other treatments. Also, VEGF and apoptotic tumor cells in the tumor tissues were detected after infusion of the cells into the mice. The results showed that the VEGF-positive rate was lowest and the number of apoptotic cells was highest in the CD/ES-EPCs group. Interpretation: The EPCs transfected with CD/ES inhibited tumor growth and preferentially induced tumor cell apoptosis. Thus, this treatment strategy might open a novel avenue for cancer-targeting therapy. Funding Statement: The present work was funded by Zhejiang Provincial Natural Science Foundation of China (Grant No.LZ18H180001), National S&T Major Project of China (NO.2018ZX10301201)，National Natural Science Foundation of China (Grant No.81371658), The Key Research Development Program of Zhejiang province (Grant No.2018C03018) and Key Science and Technology Program of Zhejiang province(No.WKJ-ZJ-1923). Declaration of Interests: The authors declare no potential conflict of interest. Ethics Approval Statement: All experimental procedures of the study were approved by our Institutional Animal Use and Care Committee. Mouse hepatocarcinoma model, aged 6 weeks, was provided by the Experimental Animal Center, College of Medicine, Zhejiang University. All experiments in this study were performed in agreement with the Guidelines for the Welfare of Animals in Experimental Neoplasia and approved our Institutional Animal Use and Care Committee.

Interface trap can act as the generation center in device to induce a very weak generation current. We observed the negative differential resistance NDR of this generation current ID in nMOSFET with the floating source. It originates from that the generation function of interface trap is enabled and then is shut down in turn as increasing the drain voltage. This change relies on the interaction among the interface trap energy-level and the electron's Fermi-levels of drain and source under the floating source condition. It is found that the peak-to-valley ratio of ID is beyond 30.

One new Zn(Ⅱ) metal–organic framework, [Zn3(tib)2(BPT)2]·7H2O (ZZIT-1) [ZZIT-1 = Zhengzhou Institute of technology, tib = 1,3,5-tris(1-imidazolyl)benzene, H3BPT = biphenyl-3,4′,5-tricarboxylic acid], has been synthesized and characterized. ZZIT-1 is a two-fold interpenetrating 3D framework structure. The gas/vapor adsorption behavior and luminescence of the complex have been studied. Notably, ZZIT-1 can selectively adsorb CO2 over N2, and MeOH over EtOH, and these adsorption results have been corroborated by theoretical calculations. Most interestingly, ZZIT-1 shows unique selectivity for the detection of acetone through a fluorescence quenching mechanism in the solid state at room temperature.

A numerical study based on the ECOMSED model was carried out to investigate the storm surge induced by super typhoon 5612 (WANDA) along Zhejiang coast, China. The modeled results presented good agreement with the observed data. According to the 70% probability circle method, 42 tracks were designed based on WANDA with various landfall locations and central pressures. Subsequently, overtopping probabilities of seawalls corresponding to each track were achieved. The results showed that variation of storm surge ranged from −0.04 to 0.04 m as the pressure increased/decreased by 5 hPa. The storm surge was easily influenced by the landfall location change with the same central pressure. The comparison of peak storm surge between modeled results and observed data at multiple stations were also illustrated. Moreover, in case of WANDA travelling within the 70% probability circle, seawalls between southern Taizhou and Wenzhou are faced with small effect; while seawalls located at southern Zhoushan and eastern Ningbo will be vulnerable and even suffered severe overtopping risk.

Chemical exfoliation method was applied to transform bulky assemblies of Au(I)–3-mercaptopropionate (MPA) coordination polymer (CP) to nanosheets and nanostrings using sodium citrate as an exfoliator. The exfoliation process and the structural characteristics of the Au(I)–MPA nanosheets and nanostrings were fully investigated by transmission electron microscopy, atomic force microscopy, UV–vis absorption spectroscopy, X-ray photoelectron spectroscopy and so on. As the structural rigidity and stability of the obtained Au(I)–MPA nanosheets, they are ideal precursors for fabrication of water soluble gold nanoparticle assemblies through progressive pyrolysis. This work provides a significant strategy toward the morphology regulation of CP nanostructures and will inspire further development of this research area.

Ethnopharmacological relevance Extracts from Ginkgo biloba L. leaves confer their therapeutic effects through the synergistic actions of flavonoid and terpenoid components, but some non-flavonoid and non-terpenoid components also exist in this extract. In the study of this paper, an investigation was carried out to compare the pharmacokinetic parameters of fourteen compounds to clarify the influences of non-flavonoid and non-terpenoid fraction (WEF) on the pharmacokinetics profile of the flavonoid fraction (FF) and the terpene lactone fraction (TLF) from Ginkgo biloba extracts. Materials and methods A selective and sensitive UPLC–MS/MS method was established to determine the plasma concentrations of the fourteen compounds to compare the pharmacokinetic parameters after orally administration of FF, TLF, FF–WEF, FF–TLF, TLF–WEF and FF–TLF–WEF with approximately the same dose. At different time points, the concentration of rutin (1), isoquercitrin (2), quercetin 3-O-[4-O-(-β-D-glucosyl)-α-L-rhamnoside] (3), ginkgolide C (4), bilobalide (5), quercitrin (6), ginkgolide B (7), ginkgolide A (8), luteolin (9), quercetin (10), apigenin (11), kaempferol (12), isorhamnetin (13), genkwanin (14) in rat plasma were determined and main pharmacokinetic parameters including T1/2, Tmax, Cmax and AUC were calculated using the DAS 3.2 software package. The statistical analysis was performed using the Student׳s t-test with P Results FF and WEF had no effect on the pharmacokinetic behaviors and parameters of the four terpene lactones, but the pharmacokinetic profiles and parameters of flavonoids changed while co-administered with non-flavonoid components. It was found that Cmax and AUC of six flavonoid aglycones in group FF–WEF, FF–TLF and FF–TLF–WEF had varying degrees of reduction in comparison with group FF, especially in group FF–TLF–WEF. On the contrary, the values of Cmax, Tmax and AUC of four flavonoid glycosides in group FF–TLF–WEF were significantly increased compared with those in group FF. Conclusions These results indicate that non-flavonoid components in Ginkgo biloba extracts could increase the absorption and improve the bioavailability of flavonoid glycosides but decrease the absorption and reduce the bioavailability of flavonoid aglycones.

Nicotinamide adenine dinucleotide (NAD(+)) plays critical roles in not only energy metabolism and mitochondrial functions, but also calcium homeostasis and immunological functions. It has been reported that NAD(+) administration can reduce ischemic brain damage. However, the mechanisms underlying the protective effects remain unclear. Because mitochondrial impairments play a key role in the cell death in cerebral ischemia, in this study we tested our hypothesis that NAD(+) can decrease mitochondrial damage-induced cell death using differentiated PC12 cells as a cellular model. We found that NAD(+) can decrease both early-stage and late-stage apoptosis, as well as necrosis of rotenone-treated PC12 cells, as assessed by FACS-based Annexin V/AAD assay. We also found that NAD(+) treatment can restore the intracellular NAD(+) levels of the rotenone-treated cells. Moreover, NAD(+) treatment can prevent rotenone-induced mitochondria depolarization. In summary, our study has provided first direct evidence that NAD(+) treatment can prevent rotenone-induced apoptosis and necrosis. Our study has also indicated that NAD(+) treatment can prevent mitochondrial damage-induced cell death, which may at least partially result from its protective effects on rotenone-induced mitochondrial depolarization. Because both mitochondrial damage and apoptosis play key roles in multiple neurological disorders, our study has highlighted the therapeutic potential of NAD(+) for brain ischemia and other neurological diseases.

Neuronal fractalkine acts via its receptor, CX3CR1, on microglia to regulate neuroinflammation. Conflicting results have been reported in studies employing CX3CR1 deficient ( Cx3cr1 −/− ) mice. Here, compared to wild-type, endotoxin-treated neuron-glial Cx3cr1 −/− cultures produced less TNF-α, nitric oxide and superoxide; however, fractalkine treatment inhibited the release of pro-inflammatory factors in wild-type and BV-2 cell cultures. Furthermore, endotoxin-treated BV-2 cells expressing siRNA against CX3CR1 increased nitric oxide and TNF-α production. We hypothesize that CX3CL1-CX3CR1 signaling is neuroprotective and propose that the reduced production of pro-inflammatory signals in Cx3cr1 −/− microglia may result from compensatory mechanisms and not be the direct result of CX3CR1 deficiency.

Emerging evidence has indicated microRNAs are involved in tumor development and progression, acting as tumor suppressors or oncogenes. Here we report that miR-409-3p was significantly downregulated in gastric cancer (GC) cell lines and tissues. Overexpression of miR-409-3p in SGC-7901 gastric cancer cells dramatically suppressed cell proliferation and induced cell apoptosis both in vitro and in vivo. Furthermore, we demonstrate that the transcriptional regulator PHF10 was a target of miR-409-3p. Taken together, these findings suggest that miR-409-3p may function as a novel tumor suppressor in GC and its anti-oncogenic activity may involve the direct targeting and inhibition of PHF10.

CD38 is a multifunctional enzyme that can not only generate cyclic adenosine diphosphate-ribose (cADPR) - a key Ca(2+) -mobilizing second messenger - by consuming NAD(+), but also hydrolyze extracellular NAD(+). There have been only a small number of studies on the functions of CD38 in the CNS. Brain inflammation plays critical roles in ischemic brain injury and multiple other neurological diseases, in which microglia activation is a key event. In this study we determined the roles of CD38 in the basal survival of mouse BV2 microglia cells by applying CD38 siRNA. Our study found that silencing of CD38 led to significantly decreased survival of the cells. We also found that decreased CD38 levels can lead to apoptosis of the microglial cells, as assessed by flow cytometry-based Annexin V/7-AAD assay, caspase-3 immunostaining and Hoechst staining assays. Our study has further indicated that the CD38 silencing-induced apoptosis is mainly caspase 3-dependent. Collectively, our study has provided the first evidence suggesting that CD38 plays a critical role in the basal survival of microglia, and decreased CD38 can lead to caspase 3-dependent apoptosis of the cells. These results suggest that CD38 may become a therapeutic target for modulating microglial survival in neurological diseases.

SIRT2 is a tubulin deacetylase, which can play either detrimental or beneficial roles in cell survival under different conditions. While it has been suggested that reduced SIRT2 expression in human gliomas may contribute to development of gliomas, there has been no study that directly determines the effects of decreased SIRT2 activity on the survival of glioma cells. In this study we applied both pharmacological and molecular approaches to determine the roles of SIRT2 in the survival of glioma cells. Our studies, by conducting such assays as flow cytometry-based Annexin V assay and caspase-3 immunostaining, have indicated that decreased SIRT2 activity leads to apoptosis of C6 glioma cells by caspase-3-dependent pathway. Our experiments have further shown that reduced SIRT2 activity produces necrosis of C6 glioma cells. Moreover, our study applying SIRT2 siRNA has also shown that decreased SIRT2 leads to both necrosis and apoptotic changes of C6 glioma cells. Collectively, our study has provided novel evidence indicating that SIRT2 activity plays a key role in maintaining the survival of glioma cells, and that reduced SIRT2 activity can induce both necrosis and caspase-3-dependent apoptosis of C6 glioma cells. These results have also suggested that inhibition of SIRT2 might become a novel therapeutic strategy for gliomas.

A nested Ecomsed model with high resolution grid has been developed and used to simulate typical typhoon storm surge (0608 typhoon) which landed on the coast of Zhejiang. Two main pressure modes have been considered in this model: Jelesnianski65 (J65) and Fujita &Takahashi (FJ) model. This study found that FJ model is agree better with the measured values than J65 model. The model has been used to simulate the water level which has been compared with the available field observations (Kanmen; Wenzhou; and Dachen stations). The results show that the error of the peak water level is about 50-70 cm in the first domain and the nested model improves the accuracy which is around 30–50cm(under 30 cm during ebb). The relative error is lower and the time effect is more accurate in the nested small domain. Thus, it is suggested that to simulate and predict typhoon storm surge a nested Ecomsed be adopted.

Hepatitis B virus (HBV) precore G1896A mutation is associated with Hepatitis B e antigen (HBeAg) seroconversion. This mutation and the adjacent G1899A mutation also appear to associate with increased risk of hepatocellular carcinoma. Quantitative mutant dynamics may help determine the potential of these mutants as clinical biomarkers. However, a reliable method to quantify either mutant is not available, partly because the viral genome has polymorphisms in general and the precore mutations are complex.(1) To develop a reliable and ultrasensitive assay for the quantification of HBV G1896A and/or G1899A mutants. (2) To obtain preliminary data on the quantities of the precore mutants in patients.A SimpleProbe real time PCR assay was developed to quantify the HBV precore mutants. Dual melting analysis and a primer-probe partial overlap approach were used to increase detection accuracy. A wild-type selective PCR blocker was also developed to increase mutant detection sensitivity.The assay correctly identified the precore sequence from all 62 patient samples analyzed. More than 97% of precore sequences in the GenBank can be recognized. Mutant detection sensitivity reached 0.001% using a wild type-selective PCR blocker. At least one precore mutant can be detected from all 20 HBeAg-positive individuals who were negative for precore mutations by DNA sequencing.The reliability of this ultrasensitive mutation quantification assay was demonstrated. The same approaches may be useful for the detection of other clinically significant mutations. Evolution of the precore mutants warrants further studies.

Previous transgenic-reporter and targeted-deletion studies indicate that the subset-specific expression of CD8αβ heterodimers is controlled by multiple enhancer activities, since no silencer elements had been found within the locus. We have identified such a silencer as L2a, a previously characterized ∼220 bp nuclear matrix associating region (MAR) located ∼4.5 kb upstream of CD8α. L2a transgenes driven by the E8 I enhancer showed no reporter expression in thymic subsets or T cells in splenic, inguinal and mesenteric lymph node peripheral T cells. Deletion of L2a resulted in significant reporter de-repression, even in the CD4 + CD8 + double positive (DP) thymocyte population. L2a contains binding sites for two MAR-interacting proteins, SATB1 and CDP. We found that that binding of these factors was markedly influenced by the content and spacing of L2a sub-motifs (L and S) and that SATB1 binds preferentially to the L motif both in vitro and in vivo. A small fraction of the transgenic CD8 single positive (SP) thymocytes and peripheral CD8 + T cells bypassed L2a-silencing to give rise to variegated expression of the transgenic reporter. Crossing the L2a-containing transgene onto a SATB1 knockdown background enhanced variegated expression, suggesting that SATB1 is critical in overcoming L2a-silenced transcription.

Controlled modulation of T-cell response during immunotherapy, especially the balance between T helper 1 (Th1) and Th2 responses, is critical for generating effective immune response. Here we report that dual delivery of interleukin 10 (IL-10)-targeted small interfering RNA (siRNA) and DNA vaccines to dendritic cells (DCs), using a single particle carrier, efficiently enhances immune response and modulates it toward a stronger Th1 phenotype. Surface-functionalized polymer microparticles (MPs) carrying both IL-10-targeted siRNA and DNA antigens exhibited effective gene silencing, DNA transfection, and synergistically enhanced upregulation of maturation markers in primary DCs leading to increased T-cell proliferation, in vitro. Mice immunized with these dual-delivery carriers demonstrated a significant "switch" toward Th1 response as evidenced by increase in interferon gamma (IFN-gamma) production and decrease in IL-4 production by CD4+ T cells. This further led to enhanced antiviral cytotoxic T-lymphocyte activity. Such dual siRNA-DNA delivery provides a novel strategy to precisely control the type and strength of T-cell response during immunotherapy.

Intrathymic signals induce the differentiation of immature CD4 + CD8 + double positive (DP) thymocytes into mature CD4 + or CD8 + single positive (SP) T cells. The transcriptional mechanism by which CD8 lineage is determined is not fully understood. The best evidence, which favors the kinetic signaling/coreceptor reversal model, indicates that signaled DP thymocytes terminate CD8 transcription prior to their subsequent re-initiation of CD8 transcription and ultimate differentiation into CD8SP T cells. We and others have shown that CD8 lineage commitment is severely perturbed in mice in which expression of the transcription factor SATB1 is either conventionally knocked out or T cell-specifically knocked down. Here, we demonstrate that, as with normal thymocytes, cultured SATB1-deficient DP thymocytes inactivate CD8 coreceptor transcription following receipt of signals (PMA plus ionomycin) that mimic TCR-mediated positive selection. However, this terminated CD8 transcription is not re-initiated by signals (IL-7) conducive to CD8 differentiation in SATB1-deficient DP. We show that SATB1 specifically binds to a cis -regulatory element within the CD8 enhancer (E8 III ) known to be required for coreceptor reversal. A requirement in CD8 coreceptor reversal identifies SATB1 as an essential trans -regulator of CD8 lineage fate, whose action may be mediated via recruitment to the E8 III DP enhancer.

A single-sided heated graphite cylindrical electrode (ss-HGCE) was designed. Compared to previous alternative current (AC) heating, much simpler and cheaper direct current (DC) heating supplier was adopted for the first time to perform adsorptive accumulation of rutin at ss-HGCE at elevated electrode temperature. This offers great promise for low cost, miniaturization and high compatibility with portability. The square wave voltammetry (SWV) stripping peak current was enhanced with increasing the electrode temperature only during preconcentration step. This enhancement was contributed to the forced thermal convection induced by heating the electrode rather than the bulk solution, which is able to improve mass transfer and facilitate adsorption hence enhance stripping response. A detection limit of 1.0 × 10−9 M (S/N = 3) could be obtained at an electrode temperature of 48 °C during 5 min accumulation, one magnitude lower than that at 28 °C (room temperature). This is the lowest value at carbon-based electrodes for rutin determination as we know. Such novel method was also successfully used to determine rutin in pharmaceutical tablets.

In this study, a two-stage fuzzy robust integer programming (TFRIP) method has been developed for planning environmental management systems under uncertainty. This approach integrates techniques of robust programming and two-stage stochastic programming within a mixed integer linear programming framework. It can facilitate dynamic analysis of capacity-expansion planning for waste management facilities within a multi-stage context. In the modeling formulation, uncertainties can be presented in terms of both possibilistic and probabilistic distributions, such that robustness of the optimization process could be enhanced. In its solution process, the fuzzy decision space is delimited into a more robust one by specifying the uncertainties through dimensional enlargement of the original fuzzy constraints. The TFRIP method is applied to a case study of long-term waste-management planning under uncertainty. The generated solutions for continuous and binary variables can provide desired waste-flow-allocation and capacity-expansion plans with a minimized system cost and a maximized system feasibility.

Glutamatergic synaptic transmission is a dynamic process determined by the amount of glutamate released by presynaptic sites, the clearance of glutamate in the synaptic cleft, and the properties of postsynaptic glutamate receptors. Clearance of glutamate in the synaptic cleft depends on passive diffusion and active uptake by glutamate transporters. In this study, we examined the role of glial glutamate transporter 1 (GLT-1) in spinal sensory processing. Excitatory postsynaptic currents (EPSCs) of substantia gelatinosa neurons recorded from spinal slices of young adult rats were analyzed before and after GLT-1 was pharmacologically blocked by dihydrokainic acid. Inhibition of GLT-1 prolonged the EPSC duration and the EPSC decay phase. The EPSC amplitudes were increased in neurons with weak synaptic input but decreased in neurons with strong synaptic input upon inhibition of GLT-1. We suggest that presynaptic inhibition, desensitization of postsynaptic AMPA receptors, and glutamate "spillover" contributed to the kinetic change of EPSCs induced by the blockade of GLT-1. Thus, GLT-1 is a key component in maintaining the spatial and temporal coding in signal transmission at the glutamatergic synapse in substantia gelatinosa neurons.

Bis(7)-tacrine [bis(7)-tetrahydroaminacrine] acts as an AChE inhibitor and also exerts modulatory effects on many ligand-gated ion channels and voltage-gated Ca(2+) and K(+) channels. It has been reported previously that tacrine and some other AChE inhibitors suppressed I(K(A)) in central and peripheral neurons. The present study aimed to explore whether bis(7)-tacrine could modulate the function of native delayed rectifier potassium channels in DRG neurons and K(V)1.2 encoded potassium channels expressed in oocytes. We found that both delayed rectifier potassium currents (I(K(DR))) in rat DRG neurons and the currents recorded from oocytes expressing K(V)1.2 (I(K(K(V)1.2))) were suppressed by bis(7)-tacrine, the potency of which was two orders greater than that of tacrine. The IC(50) values for bis(7)-tacrine and tacrine inhibition of I(K(KD)) in DRG neurons were 0.72+/-0.05 and 58.3+/-3.7 microM, respectively; while the two agents inhibited I(K(K(V)1.2)) in oocytes with an IC(50) of 0.24+/-0.06 and 102.1+/-21.5 microM, respectively. The possible mechanism for bis(7)-tacrine inhibition of I(K(A)) and I(K(K(V)1.2)) was identified as the suppression of their activation, inactivation.

In this paper, L- and W-surface are constructed via recurrence scheme. The derivation formulae, basis function properties, envelope theorems of L-surface and an equivalent representation of L-surface into Bezier surface are presented. Especially, a C2 bicubic spline surface on a cross-cut grid partition consisting of quadrilaterals is given, which can be used to replace the biquintic surface in most of the applications. The new method has been applied in the field of contour design of human body and aircraft.