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Curcumin is a natural herbal product that has been popularly used to treat autoimmune diseases in China; however, its effects on rheumatoid arthritis and its mechanism are not clear. The main purposes of this study are to explore the therapeutic effects of curcumin on collagen-induced arthritis (CIA) rats and the pharmacological mechanism. In the present study, CIA rats were established by injecting bovine type II collagen. Curcumin and methotrexate were then orally administered daily, and the swelling degree of the hind limb joints was scored every two days. Histopathological changes were observed by hematoxylin-eosin staining. The levels of cytokines (TNF-α, IL-1β, IL-17 and TGF-β) were detected by radioimmunoassay, while the expression of IκBα and COX-2 was detected by Western blot. In addition, cell viability was detected by CCK-8 assay, and the effect of curcumin on macrophage apoptosis was detected by flow cytometry and TUNEL assay. The results indicated that in vivo curcumin attenuated the degree of joint swelling of rats and the further development of joint histopathology. Moreover, it downregulated the levels of cytokines. In vitro curcumin inhibited the degradation of IκBα and reduced the production of COX-2 in LPS-induced inflammatory RAW264.7 cells. Importantly, curcumin significantly induced macrophage apoptosis. In conclusion, in this study, we have demonstrated that curcumin exerts therapeutic effects on arthritis in CIA rats and has a strong pharmacological activity on reducing the inflammatory response in macrophages. Its mechanism may be related to the inhibition of the NF-κB signaling pathway and the promotion of macrophage apoptosis.

[This corrects the article DOI: 10.1016/j.cytox.2019.100015.].

The friend leukemia integration 1 (Fli-1) gene is involved in the expression control of key genes in multiple pathogenic/physiological processes, including cell growth, differentiation, and apoptosis, this implies that Fli-1 is a strong candidate for drug development. In our previous study, a 3′,5′-diprenylated chalcone, (E)-1-(2-hydroxy-4-methoxy-3,5-diprenyl) phenyl-3-(3-pyridinyl)-propene-1-one (C10), was identified as a novel anti-prostate cancer (PCa) agent. Here, we investigated the molecular mechanisms underlying the anti-cancer effects of C10 on the growth, metastasis, and invasion of PC3 cells in vitro. Our results show that C10 exhibited a strong inhibitory effect on proliferation and metastasis of PC3 cells via several cellular and flow cytometric analyses. Further mechanism studies revealed that C10 likely serves as an Fli-1 agonist for regulating the expression of Fli-1 target genes including phosphatidylinositol 3-kinase (P110), murine double minute2 (MDM2), B-cell lymphoma-2 (Bcl-2), Src homology-2 domain-containing inositol 5-phosphatase 1 (SHIP-1), and globin transcription factor-1 (Gata-1) as well as the phosphorylation of extracellular-regulated protein kinases 1 (ERK1). Further, we confirmed that C10 can regulate the expressions of vascular endothelial growth factor 1 (VEGF-1), transforming growth factor-β2 (TGF-β2), intercellular cell adhesion molecule-1 (ICAM-1), p53, and matrix metalloproteinase 1 (MMP-1) genes associated with tumor apoptosis, migration, and invasion. Thus, C10 exhibits stronger anticancer activity with novel molecular targets and regulatory molecular mechanisms, indicating its great potency for development as a novel targeted anticancer drug.

Although microRNA (miR)-145 has been identified to be a tumor suppressor in various types of tumor, it promotes the progression of non-small cell lung cancer (NSCLC). However, the precise underlying molecular mechanism of its action remains unclear. The present study investigated the effects of miR-145 on the proliferation, invasion, metastasis and apoptosis of the NSCLC A549 cell line and the underlying molecular mechanism of its action. In vitro cell proliferation, invasion, migration and apoptosis assays were employed, and the expression levels of matrix metalloproteinase (MMP)-2, MMP-9, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), caspase-3and poly(ADP-ribose) polymerase (PARP) were evaluated by western blot analysis. The results demonstrated that ectopic expression of miR-145 inhibited the proliferation, invasion and migration of A549 cells, but promoted the apoptosis of A549 cells. Western blot analysis indicated that increased miR-145 levels led to a marked decrease in the expression of MMP-2, MMP-9 and Bcl-2. Upregulation of miR-145 expression increased the expression of Bax, thus increasing the Bax/Bcl-2 ratio. Additionally, the results indicated that miR-145 over expression promoted the cleavage of caspase-3 and PARP. Taken together, these results indicated that miR-145 suppresses the proliferative, invasive and migratory ability of A549 cells. Additionally, miR-145 upregulation induced apoptosis of A549 cells possibly by decreasing MMP-2 and MMP-9 expression, the Bax/Bcl-2 ratio and the activity of the caspase-3 cascade.

Due to their possible ideal three dimensional (3D) nanostructures and excellent electrochemical properties, conducting hydrogels have attracted much attention in recent years. Herein, pyrrole monomer was directly dissolved in an aqueous solution of sodium alginate (SA), and was allowed to undergo in-situ polymerization to form polypyrrole (Ppy), resulting in formation of Ppy/SA, a conducting hydrogel, via self-assembly between the polymers. Observation by SEM indicated that the microstructure of the Ppy/SA hydrogel was a typical 3D nano-cylinder network, with the cylinders formed by entanglements of the Ppy and SA molecular chains (cylinder diameter was ∼100 nm). The electrochemical measurements of cyclic voltammetry, galvanostatic charge/discharge and electrochemical impedance spectroscopy indicated that the Ppy/SA hydrogel possessed typical pseudocapacitance, good charging and discharging rate performance, and favorable capacitive behavior; the specific capacity reached up to 249 F/g at the curr...

A double Claisen rearrangements synthetic strategy was established for the total synthesis of 4,4′-dimethyl medicagenin (compound 6c). A series of its analogs also were prepared, including two novel 3′,5′-diprenylated chalcones, in which ring B was replaced by azaheterocycle. The structures of the twenty-two newly synthesized compounds were confirmed by 1H NMR, 13C NMR and ESI-MS. In vitro, the cytotoxicity of the target compounds was evaluated using cancer cells. Noticeably, compound 10 exhibited broad-spectrum cytotoxicity on PC3 prostate cancer cells, MDA-MB-231 breast cancer cells (MDA), HEL and K562 erythroleukemia cells with IC50 values of 2.92, 3.14, 1.85 and 2.64 μM, respectively. Further studies indicated that compound 10 induced apoptosis and arrested the cell cycle phase of the above mentioned four cancer cell lines. By contrast, compound 6g selectively displayed potent inhibitory activity against the proliferation of HEL cells with an IC50 value of 4.35 μM. Compound 6g slightly induced apoptosis and arrested cell cycle phase of HEL cells. Preliminary structure-activity relationship studies indicated that, in all cancer cell lines evaluated, the 3-pyridinyl group was essential for cytotoxicity.

Conducting hydrogels were obtained through high-pressure induced gelation of poly(vinyl alcohol) aqueous solutions and subsequent in-situ polymerization of aniline (polyaniline). A study of scanning electron microscopy showed that polyaniline was homogeneously dispersed within the 3D nano-network matrix of the poly(vinyl alcohol) hydrogels. The UV–vis spectra and cyclic voltammetry of the hydrogels demonstrated that the high-pressure treatment had no influence on the in-situ polymerization and electrochemical feature of polyaniline. A study of electrochemical impedance indicated that the conducting hydrogels possessed electrical conductivity and potential application in the field of biomaterials and electrode materials of supercapacitors.

Previous studies suggest that radiotherapy (RT) can induce immune activation, which not only reduces the progression of tumors, but also causes the release of tumor antigens. The combination of RT and immune checkpoint blockade, such as the inhibition of programmed cell death 1 (PD‑1) and programmed cell death ligand 1 (PD‑L1), has been demonstrated to yield impressive response rates. However, a substantial proportion of patients develop resistance such therapies. Previous studies have shown that indoleamine 2,3‑dioxygenase (IDO) causes T cell exhaustion and increased formation of regulatory T cells (Tregs), upregulating the expression of inhibitory receptors and ligands. Therefore, the application of IDO inhibitors combined with RT may have a synergistic effect by relieving immunosuppression. Lewis lung cancer cell‑bearing mice were treated with the IDO inhibitor 1‑methyl‑tryptophan (1MT) and/or 10 Gy RT. Tumor size was measured every day, flow cytometry was performed to measure the expression of dendritic cell (DC) maturation markers, inhibitory receptors, ligands, cytotoxic T cells and Treg phenotypic markers. Reverse transcription‑quantitative PCR was used to evaluate the mRNA expression levels of IDO, PD‑L1, PD‑1, T cell immunoglobulin domain and mucin domain 3 (TIM‑3), B‑ and T‑lymphocyte attenuator (BTLA) and galectin‑9. Compared with the control group, treatment with 1MT or RT reduced tumor growth, however, the combination therapy was more effective than either treatment alone. Flow cytometry showed the upregulation of CD80, CD86 and the major histocompatibility complex II in spleen DCs and the concurrent downregulation of CD4, CD25 and forkhead box protein P3 in lymphocytes in the treatment groups. Compared with the control group, inhibitory receptors and ligands that affect DCs and T cells were observed, expression levels of PD‑L1, PD‑1, TIM‑3, BTLA and galectin‑9 are decreased in treatment group compared with control. IDO inhibition synergized with RT to downregulate Tregs, inhibitory receptors and ligands to prevent T cell exhaustion. By activating DCs and T cells, this combination therapy may strongly enhance antitumor immunity and inhibit tumor progression.

In the present work, by using the in situ polymerization of aniline in acidic aqueous solution of PVA and freezing–thawing method, the reinforced conducting polymer hydrogel of polyaniline/polyvinyl alcohol (PANI/PVA) was prepared and directly used as the self-supported electrode for supercapacitor. The chemical structure of PANI/PVA hydrogel was characterized by UV–Vis, and the three-dimensional (3D) interconnected hierarchical nanoporous structure was observed by scanning electron microscopy. The measurement of compressive strength demonstrated the high mechanical strength of the hydrogel. The electrochemical properties of PANI/PVA hydrogel electrodes were evaluated by using cyclic voltammetry, electrochemical impedance spectroscopy and galvanostatic charge/discharge, which indicated their good responsiveness and rate capability, low resistance, high specific capacitance (240 F/g at current density of 1 A/g) and excellent cycling stability. The easily fabricated PANI/PVA hydrogel combined hierarchical nanoporous microstructure, self-supported feature and favorable capacitive behavior and provided a new strategy for constructing high-performance electrode for supercapacitors.

The functionalization of halloysite nanotubes (HNTs) has attracted much attention in recent years due to the nano-tubular structure and particular features. Here, we report a conductive nanohybrids of HNTs, polyaniline (PANI), and poly(sodium-p-styrenesulfonate) (PSS) with high electrical conductivity (0.11 S/cm) prepared by in situ polymerization and layer-by-layer assembly. The chemical structures and morphologies were characterized by FT-IR, UV–Vis, FE-SEM, and TEM, which confirmed that the layers of PANI, PSS, and PANI were sequentially coated onto the HNTs as expected. A study of cyclic voltammetry suggested the typical pseudocapacitance and good rate performance of the as-prepared nanocomposites. The results of galvanostatic charge/discharge and electrochemical impedance spectroscopy further demonstrated its favorable capacitive behavior and low resistance. The easily fabricated halloysite/polyaniline nanocomposites show great potential as electrode materials for supercapacitors.

Glutathione–Ag (GSH–Ag) supramolecular hydrogels have almost no mechanical strength, and the disassembly of GSH–Ag complexes occurs very easily under UV irradiation. In this work, low molecular weight polyvinyl alcohol (PVA) was used to reinforce GSH–Ag gels. The phase transition temperature of GSH–Ag gels can be increased from 45 to 60 °C in the presence of only 0.8 wt% of PVA. Rheological studies indicated that the storage modulus of reinforced GSH–Ag gels was 6 times higher than that of unreinforced gels. SEM studies revealed that the macromolecular chains of PVA are intertwined with the GSH–Ag aggregates, leading to an increase of mechanical strength. The TEM studies and UV-vis spectra indicated that Ag+ within the GSH–Ag gels was in situ reduced to form Ag nanoparticles under UV irradiation. Meanwhile, gradual disassembly of GSH–Ag hydrogels can be induced through controlling the time of UV irradiation. Based on this feature, the controllable release of a loaded model drug (crystal violet) and Ag species can be realized from the reinforced GSH–Ag gels. The release media show a good inhibition effect on E. coli.

Multiple geophysical parameters including groundwater level, water temperature, water radon, crustal deformation, electromagnetic waves and total electron contents (TEC) in the ionosphere, are examined together to investigate pre-earthquake anomalous phenomena associated with the Mw6.1 Ludian earthquake on August 3, 2014. Cross-parameter comparison eliminates anomalies that are detected in one parameter only and examine earthquake-related phenomena in various fields, simultaneously. Retrieved anomalies show that abnormal decreases of water temperature and radon concentration related to anomalous uplift of groundwater levels appeared about 1–8 days before the earthquake even without contributions from rain. Significant step-like changes are observed from data recorded by strainmeters on July 31 and are consistent with water-related anomalies in timing. No significant anomalies can be found from electromagnetic data through the preliminary observation. In contrast, TEC appeared positive anomalies 1–3 days before the Ludian earthquake. Although causal mechanisms of the TEC anomalies from stressed rocks are not fully understood, multiple-parameter examination can increase credibility of determination regarding pre-earthquake phenomena.

Rheumatoid arthritis (RA) is a chronic, symmetric, systemic autoimmune disease. Because insufficient apoptosis of fibroblast-like synoviocytes (FLS) is an important characteristic of RA, promoting apoptosis is considered a potential therapeutic tool for treating RA. We have previously found that daphnetin (7,8-dihydroxycoumarin, DAP) has a pro-apoptotic effect on fibroblast-like synoviocytes from collagen-induced arthritis (CIA) rats. In the present study, we further investigated the mechanisms of DAP-induced apoptosis in CIA-FLS. CIA-FLS were incubated with DAP for 48 h in the presence or absence of caspase inhibitors, including inhibitors of caspase-3, caspase-8, or caspase-9 or a pan-caspase inhibitor; then, a series of experiments were performed to evaluate the mechanisms of DAP-induced apoptosis. Our results showed that DAP markedly decreased cell viability and induced the apoptosis of CIA-FLS along with typical morphological and ultrastructural changes; moreover, DAP increased FasL, cytochrome c (Cyt-c), Bax, caspase-3, caspase-8, and caspase-9 mRNA expression and Bax, caspase-3, caspase-8, and caspase-9 protein expression. In contrast, DAP decreased Bcl-2 mRNA and protein expression and promoted the release of Cyt-c from the mitochondria into the cytosol; these effects were attenuated to varying degrees by pre-treatment with caspase inhibitors, especially with caspase-3 or caspase-9 inhibitors or a pan-caspase inhibitor. In conclusion, the current findings demonstrate that the DAP-induced apoptosis of CIA-FLS occurred mainly via a caspase-dependent pathway, in particular the mitochondrial pathway, and that the Bax/Bcl-2 ratio was involved in this process. Thus, DAP may be a potential therapeutic agent for RA.

Daphnetin (DAP), an active ingredient extracted from Daphne odora, has pharmacological effects such as anti-inflammatory, antioxidation and anti-tumor properties. The current study aims to investigate the relationship between the anti-rheumatoid effect of DAP and the inhibition of both the PI3K/AKT/mTOR and autophagy signaling pathways. DAP inhibited the proliferation of CIA-FLS in a dose-dependent manner and induce apoptosis, accelerated the G1/G0 phase and inhibited the S phase. DAP reduced the phosphorylation of AKT and mTOR and the expression of Atg5, Beclin-1 and LC3-II/LC3-I in CIA-FLS induced by TNF-α. DAP also reduced the inflammatory response in CIA-FLS induced by TNF-α by inhibiting the cytokine expression of TNF-α, IL-6, TGF-β, IL-17, and INF-γ and promoting IL-10 expression. Overall, DAP inhibited the proliferation of CIA-FLS by down-regulating the PI3K/AKT/mTOR signaling pathway and inhibited autophagy in order to induces apoptosis, which may be potential therapeutic approach in treatment of RA.

Surgical and medical treatments usually fail to completely remove the primary lesions of salivary adenoid cystic carcinoma (SACC), resulting in local recurrence due to its strong infiltration, hematogenous metastasis and other unique biological behaviors. Targeted gene therapy, including hypoxia‑inducible factor‑1α (HIF‑1α) therapy, sheds new light on the treatment of salivary gland tumors. However, the mechanisms underlying the downregulation of HIF‑1α expression in SACC have not been well studied. The present study aimed to determine the effects of HIF‑1α downregulation on the proliferation, invasion, apoptosis, cell cycle and angiogenesis of hypoxic SACC‑83 cells, and to elucidate the molecular mechanisms underlying these effects. Western blot analysis was used to evaluate the protein expression levels of HIF‑1α and matrix metalloproteinase‑2 (MMP‑2). Angiogenesis and the protein expression of vascular endothelial growth factor (VEGF) were detected by tube formation assay of human umbilical vein endothelial cells and ELISA, respectively. It was revealed that short hairpin RNA was considerably downregulated following overexpression of HIF‑1α. In addition, downregulation of HIF‑1α inhibited the expression of VEGF and MMP‑2, as well as the proliferation, invasion, migration, and tube formation of hypoxic SACC‑83 cells, while inducing apoptosis in these cells. These results suggest that the downregulation of HIF‑1α may be a novel targeted therapy for SACC.

Leukemia is a hematologic malignancy with poor prognosis in humans and chemotherapy is the main strategy for treating leukemia patients. Novel drugs with better selectivity and lower toxicity are required for the treatment of patients. A novel 3',5'-diprenylated chalcone, (E)-1-(2-hydroxy-4-methoxy-3,5-diprenyl) phenyl-3-(3- pyridinyl)-propene-1-one (C10) is a potential new anti-leukemia agent. In this study, we investigated the molecular mechanisms of the anti-leukemia effects of C10 on different leukemia cells in vitro. C10 showed strong inhibition of proliferation of the human erythroleukemia cell line HEL and human myeloid leukemia cell line K562, and several cell and flow cytometer assays showed that inhibition by C10 was due to the regulation of gene expression or phosphorylation in the apoptosis and autophagy pathways. The results showed that C10 regulated the expression of Bax, c-Myc, Bcl-2, P38/AMPK and ERK 1/2, activated the expression of Caspase-3, -8, and PARP at the protein level in the apoptosis pathway of the two leukemia cell types, and inhibited the expression of erythroleukemia carcinogene Fli-1 in the human erythroleukemia cell line HEL. Additionally,treatment with the compound induced a time-dependent increase in expression of LC 3A/B via inhibiting the AKT-mTOR pathway, which is associated with cell autophagy. Taken together, the above results suggest that the novel synthesized 3',5'-diprenylated chalcone can prevent the growth of leukemia cells by inducing apoptosis and autophagy.

Placenta-specific 1 (PLAC1), a novel cancer-testis antigen (CTA), is expressed in a number of different human malignancies. It is frequently produced in breast cancer, serving a function in tumorigenesis. Adoptive immunotherapy using T cell receptor (TCR)-engineered T cells against CTA mediates objective tumor regression; however, to the best of our knowledge, targeting PLAC1 using engineered T cells has not yet been attempted. In the present study, the cDNAs encoding TCRα- and β-chains specific for human leukocyte antigen (HLA)-A*0201-restricted PLAC1 were cloned from a cytotoxic T-lymphocyte, generated by in vitro by the stimulation of CD8+ T cells using autologous HLA-A2+ dendritic cells loaded with a PLAC1-specific peptide (p28-36, VLCSIDWFM). The TCRα/β-chains were linked by a 2A peptide linker (TCRα-Thosea asigna virus-TCRβ), and the constructs were cloned into the lentiviral vector, followed by transduction into human cytotoxic (CD8+) T cells. The efficiency of transduction was up to 25.16%, as detected by PLAC1 multimers. TCR-transduced CD8+ T cells, co-cultured with human non-metastatic breast cancer MCF-7 cells (PLAC1+, HLA-A2+) and triple-negative breast cancer MDAMB-231 cells (PLAC1+, HLA-A2+), produced interferon γ and tumor necrosis factor α, suggesting TCR activation. Furthermore, the PLAC1 TCR-transduced CD8+ T cells efficiently and specifically identified and annihilated the HLA-A2+/PLAC1+ breast cancer cell lines in a lactate dehydrogenase activity assay. Western blot analysis demonstrated that TCR transduction stimulated the production of mitogen-activated protein kinase signaling molecules, extracellular signal-regulated kinases 1/2 and nuclear factor-κB, through phosphoinositide 3-kinase γ-mediated phosphorylation of protein kinase B in CD8+ T cells. Xenograft mouse assays revealed that PLAC1 TCR-transduced CD8+T cells significantly delayed the tumor progression in mice-bearing breast cancer compared with normal saline or negative control-transduced groups. In conclusion, a novel HLA-A2-restricted and PLAC1-specific TCR was identified. The present study demonstrated PLAC1 to be a potential target for breast cancer treatment; and the usage of PLAC1-specific TCR-engineered T cells may be a novel strategy for PLAC1-positive breast cancer treatment.

Due to the very poor mechanical strength of polyaniline-poly(styrene sulfonate) (PANI-PSS) conducting hydrogels, we explored the possibility to improve the strength of PANI-PSS hydrogels using supramolecular nanofibers formed by self-assembly of sorbitol derivatives (DBS). The compressive strength of these reinforced hydrogels was found to be 11 times higher than that of the unreinforced one. SEM studies indicated that the in-situ formed DBS nanofibers and PANI-PSS chains are entangled, leading to improvement of the hydrogel strength. The conductivity of reinforced hydrogels measured by impedance was found to be 10−4 S/cm, rather similar to that of the unreinforced hydrogels. In addition, the spectra of cyclic voltammetry for the reinforced and unreinforced hydrogels were almost the same. These results reveal that the presence of DBS nanofibers does not affect the electric properties of PANI-PSS hydrogels. As drug carriers with electric-driven release, the release rate of model drug Rhodamine B loaded within reinforced PANI-PSS hydrogels distinctly increased when an increase of voltage was applied. Furthermore, a pulse release could be realized when the voltage was alternatively applied.

Melanoma is significantly associated with mutant BRAF gene, a suitable target for siRNA-based anti-melanoma therapy. However, a tumor-specific delivery system is a major hurdle for clinical applications. Here, we developed a novel nano-carrier, FA-GNR-siBRAF for safe topical application, which consists of folic acid (FA) as the tumor-targeting moiety, golden nanorods (GNR) providing photothermal capability to kill tumor cells under laser irradiation, and siRNA specifically silencing BRAF (siBRAF). The in vitro and in vivo results revealed that FA-GNR-siBRAF displayed high transfection rates, and subsequently induced remarkable gene knockdown of BRAF, resulting in suppression of melanoma growth due to the interruption of the MEK/ERK pathway. Combinatorial photothermal effects and BRAF knockdown by FA-GNR-siBRAF effectively killed tumor cells through apoptosis, with enhanced efficiency than individual treatments. Therefore, the FA-GNR-siBRAF simultaneously induced BRAF gene silencing and photothermal effects which achieved synergistic efficacy in the treatment of melanoma, paving a new path for developing clinical treatment methods for melanoma.

Amination of tertiary and secondary alcohols using aqueous ammonia as nitrogen source was carried out by a process with recyclable intramolecular reaction of 1,8-naphthosultone, which lead to α-branched primary amines. Sulfonic resin serves as the heterogeneous catalyst for C N bond formation and protects the neighboring hydroxyl group until the required hydrolysis starts in the alkaline solution. The process can be conducted under mild conditions, no additional solvent is needed and no overreaction to secondary or tertiary amines occurs.

Novel conducting hydrogels (PVA-TA) with dual network structures were synthesized by the grafting reaction of tetraaniline (TA) into the main chains of poly(vinyl alcohol) and in situ reinforced by self-assembly of a sorbitol derivative as the gelator. The chemical structure of the PVA-TA hydrogels was characterized by using FT-IR and NMR. The mechanical strength of the PVA-TA hydrogels was strongly improved due to the presence of supramolecular nanofibers. For instance, the compressive and tensile strengths of supramolecular nanofiber-reinforced hydrogels were, respectively, 10 times and 5 times higher than those of PVA-TA hydrogels. Their storage modulus (G') and loss modulus (G″) were 5 times and 21 times higher than those of PVA-TA hydrogels, respectively. Cyclic voltammetry and conductivity measurements indicated that the electroactivity of reinforced hydrogels is not influenced by the presence of supramolecular nanofibers.

Using the PMS pressure chamber and isotope mass spectrometer (MAT-252), the leaf juice of Acacia mangium was obtained, and the carbon isotope discrimination (D) representing the most recently fixed carbon in the juice was determined. At the same time, the water-use efficiency of A. mangium was estimated. The results indicated that the carbon isotope ratio in the air of forest canopy (δa), 10 m high a bove ground averaged − 7.57 ± 1.41 ‰ in cloudy days, and − 8.54±0.67‰ in sunny days, respectively. The diurnal change of the carbon isotope ratio in the photosynthetic products of the leaf juice (δp) was of saddle type in cloudy days, but dropped down from morning to later afternoon in sunny days. A strong negative correlation between δp and leaf-to-air vapor pressure deficit (D) was observed in sunny days, but a slight change in δp was found in cloudy days. The δp also decreased with decreasing leaf water potential (Ψ), reflecting that water stress could cause the decrease of δp. The carbon isotope discrimination of the leaf juice was positively correlated with the ratio between intercellular (Pi) and atmospheric (Pa) partial pressure of CO2. For A. mangium, the isotope effect on diffusion of atmospheric CO2 via stomata was denoted by a = 4.6‰, and that in net C3 diffusion with respect to Pi was indicated by b = 28.2‰. The results were in reasonable accord with the theoretically diffusive and biochemical fractionation of carbon isotope. It was defined that carbon isotope discrimination of photosynthetic products in A. mangium leaf juice was in proportion to that from photosynthetic products in dry material. The water-use efficiency estimated by the carbon isotope discrimination in leaf juice, fit well with that measured by gas exchange system (R2 = 0.86, p < 0.0001). The application of leaf juice in measuring the stable carbon isotope discrimination would reduce the effects of fluctuating environmental factors during the synthesis of dry matter, and improve the ecophysiological studies on carbon and water balance when scaling from the plant to canopy in the fields.

CuO nanoparticle was synthesized via wet chemical method and was characterized by X-ray diffraction (XRD), nitrogen adsorption-desorption, and scanning electron microscopy (SEM). Catalytic oxidation of cumene with molecular oxygen was studied over CuO nanoparticle. The catalysts showed markedly higher activities as compared to CuO prepared by conventional method, CuO/Al2O3,orhomogeneous copper catalyst under comparable reaction conditions. The cumene conversion, cumene hydroperoxide (CHP) yield, and selectivity using 0.25 g CuO nanoparticle catalyst and 0.1 mol cumene at 358 K for 7 h were 44.2%, 41.2% and 93.2%, respectively. The catalyst can be recycled. After 6 recycled experiments, no loss of catalytic activity was observed.

The available methods for studying C uptake of forest and their problems in practices are reviewed, and a new approach to combining sap flow and 13C techniques is proposed in this paper. This approach, obtained through strict mathematic derivation, combines sap flow measurement-based canopy stomatal conductance and 13C discrimination to estimate instantaneous carbon assimilation rate of a forest. Namely the mean canopy stomatal conductance (g c) acquired from accurate measurement of sap flux density is integrated with the relationship between 13C discrimination (Δ) and Ci/Ca (intercellular/ambient CO2 concentrations) and with that between Anet (net photosynthetic rate) and g co2 (stomatal conductance for CO2) so that a new relation between forest C uptake and Δ as well as gc is established. It is a new method of such kind for studying the C exchange between forest and atmosphere based on experimental ecology.

Highlights: • Baicalin increases the ratio of the G0/G1 stages and C. albicans apoptosis. • Baicalin decreases the proliferation index of C. albicans. • Baicalin inhibits the biosynthesis of DNA, RNA and protein in C. albicans. • Baicalin depresses Succinate Dehydrogenase and Ca{sup 2+}–Mg{sup 2+} ATPase in C. albicans. • Baicalin increases the endocytic free Ca{sup 2+} concentration in C. albicans. - Abstract: Background: These experiments were employed to explore the mechanisms underlying baicalin action on Candida albicans. Methodology and principal findings: We detected the baicalin inhibition effects on three isotope-labeled precursors of {sup 3}H-UdR, {sup 3}H-TdR and {sup 3}H-leucine incorporation into C. albicans using the isotope incorporation technology. The activities of Succinate Dehydrogenase (SDH), cytochrome oxidase (CCO) and Ca{sup 2+}–Mg{sup 2+} ATPase, cytosolic Ca{sup 2+} concentration, the cell cycle and apoptosis, as well as the ultrastructure of C.albicans were also tested. We found that baicalin inhibited {sup 3}H-UdR, {sup 3}H-TdR and {sup 3}H-leucine incorporation into C.albicans (P < 0.005). The activities of the SDH and Ca{sup 2+}–Mg{sup 2+} ATPase of C.albicans in baicalin groups were lower than those in control group (P < 0.05). Ca{sup 2+} concentrations of C. albicans in baicalin groups were much higher than those inmore » control group (P < 0.05). The ratio of C.albicans at the G0/G1 stage increased in baicalin groups in dose dependent manner (P < 0.01). There were a significant differences in the apoptosis rate of C.albicans between baicalin and control groups (P < 0.01). After 12–48 h incubation with baicalin (1 mg/ml), C. albicans shown to be markedly damaged under transmission electron micrographs. Innovation and significance: Baicalin can increase the apoptosis rate of C. albicans. These effects of Baicalin may involved in its inhibiting the activities of the SDH and Ca{sup 2+}–Mg{sup 2+} ATPase, increasing cytosolic Ca{sup 2+} content and damaging the ultrastructure of C. albicans.« less

The rate of formation of the covalent thiamin diphosphate-bound enamine/C2-α-carbanion intermediate monitored at 440 nm from conjugated mechanism-based inhibitors of the structure (E)-YC 6 H 4 CH≡HCOCOOH, where Y=p-Cl, m-NO 2 , m-CF 3 , was determined in the absence and presence of the allosteric activator pyruvamide on brewers'yeast pyruvate decarboxylase (PDC, E.C. 4. 1. 1.1). For all three compounds the first-order rate constant for enamine formation was accelerated from 15-150-cold by conversion of the enzyme to its activated form

Glutathione-activated neocarzinostatin chromophore generates bistranded lesions in the hybrid formed by yeast tRNA(phe) and DNA complementary to its 31-mer 3' terminus. To elucidate the chemistry of the RNA cleavage reaction and to show that the lesions are double-stranded (ds), a series of shorter oligoribonucleotides containing the target sequence r(AGAAUUC).(GAATTCT) (underlining indicates major attack site) was studied as substrates. In addition to cleavage at both U residues, major damage was produced in the form of an abasic site at the U residues. Evidence for abasic site formation on the RNA strand was obtained from sequencing-gel analysis and measurement of uracil base release. Initial evidence for the ds nature of the damage came from experiments in which 2'-O-methyluridine was substituted for uridine in the RNA at one or both of the target sites. The site containing the substitution was not a target for cleavage or abasic site formation, and the particular T residue, staggered two nucleotides in the 3' direction on the complementary DNA strand, was cleaved significantly less. These studies were valuable in identifying the DNA ds partner of the RNA attack site. Direct evidence for ds lesions came from analysis of the products from a hairpin oligonucleotide construct in which the RNA and DNA strands were linked by four T residues and contained an internal 32P label at the 3' end of the RNA strand. Substitution of deuterium for hydrogen at the C-1' position of the U residues led to a substantial isotope effect (k1H/k2H = 3) upon the formation of the RNA abasic lesion and the RNA cleavage products, providing conclusive evidence for selective 1' chemistry. On the other hand, cleavage at the T residues on the complementary DNA strand involved C-5' hydrogen abstraction, as was also true for the T residue in an oligodeoxynucleotide analogue of the RNA strand. Chemical mechanisms to account for the RNA cleavage and abasic site formation via C-1' hydrogen abstraction are proposed.