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Cylindrospermopsin (CYN) is a toxic secondary metabolite from cyanobacteria that can cause cardiovascular disease. However, the study of CYN-induced cardiovascular toxicity in vitro is very limited and the mechanism is remain to be clarified. Vascular smooth muscle cells (VMSCs) have an important function in maintaining the structural and functional integrity of the aortic wall, and are an important in vitro model for cardiovascular research. Thus, the effects of CYN exposure (2, 20, 200, and 2000 nM) on VMSCs were analyzed. In vitro study, results showed that CYN exposure decreased VMSCs viability, inhibited VMSCs migration, induced DNA damage, destroyed cytoskeleton, changed cell morphology, promoted VMSCs apoptosis, and increased intracellular reactive oxygen species (ROS) levels. In addition, CYN could induce the activities of SOD, CAT and GPX, and promote the expressions of SOD1, CAT, GPx1, p53 and Bax genes and inhibit the expression of Bcl-2 gene, leading to a higher ratio of Bax/Bcl-2. Taken together, CYN may induce ROS overproduction, leading to increased p53 expression and ultimately promoting VSMC apoptosis. Therefore, the present study demonstrates that CYN could impair VMSCs, leading to vascular developmental defects and angiocardiopathy.

Three major polysaccharides (wHEP-1, wHEP-2, and wHEP-3) were isolated from the mycelium of Hericium erinaceus. This study assessed their chemical and physical properties, molecular weight, monosaccharide compositions, and anti-ulcerative colitis ability to protect Caco-2 cells from lipopolysaccharide (LPS)-induced inflammation. The results showed that the average molecular weight of wHEP-1, wHEP-2, and wHEP-3 was 5010, 1812, and 1118 Da, respectively. wHEP-1 was composed of mannose, glucose, and galactose in a molar ratio of 1.2:16.9:1:1, whereas wHEP-2 and wHEP-3 were composed of glucose and galactose in different molar ratios. Anti-inflammatory activity was compared using LPS-induced Caco-2 cells and a rat model. wHEP-1 exhibited the best anti-inflammatory activity. Thus, the polysaccharide from H. erinaceus as a natural material shows potential for the development of alternative therapies.

The main content in the paper aims to characterize the moisture transfer property of polytetrafluoroethylene (PTFE)/polyethylene terephthalate (PET) bi-layer complex fabric. Firstly, theoretical analyses of moisture permeability from the moisture transfer cup method and moisture resistance from the moisture resistance method were conducted, and the function between the two indexes was acquired. Results show that there is a negative correlation between moisture permeability and moisture resistance. The higher the moisture permeability of the fabric, the lower the moisture resistance. Then, in order to study relationship between moisture permeability and resistance, several PTFE/PET bi-layer complex fabrics were prepared, and a theoretical model of moisture resistance was constructed by the serial method so as to evaluate the relationship of the moisture transfer property between complex fabric and two components (PTFE film and PET fabric). Comparisons between theoretical and experimental results of moisture resistance of complex fabric were conducted, which demonstrated that the serial model was effective in characterizing the moisture transfer property of complex fabric assembled by membrane and fabric.

Flexible and durable electrically driven heaters (EDHs) was prepared by using silver nanowire/polydopamine modified cotton fabric (AgNW/PDA-C) as Joule heat generator and polydimethylsiloxane (PDMS) as stretchable matrix.The electric property of the EDHs was investigated, and it can show excellent electrical property and stability. The EDHs can be seen from the SEM morphology that cotton fabric was well protected by the PDMS and has good conductive network. The EDHs also exhibit excellent electrothermal performance under applied voltage. At low-voltages, it can quickly reach satisfactory steady-state temperatures that meet the needs of wearables. By improving the electrothermal model, it can be verified that there is a quadratic function relationship between electric power and steady-state temperature. Under cyclic voltage and bending tests, the EDHs can well maintain steady-state temperature and exhibit excellent dynamic and static electrothermal performance.

The eubacterial β sliding clamp (DnaN) plays a crucial role in DNA metabolism through direct interactions with DNA, polymerases, and a variety of protein factors. A canonical protein-DnaN interaction has been identified in Escherichia coli and some other species, during which protein partners are tethered into the conserved canonical hydrophobic crevice of DnaN via the consensus β-binding motif. Caulobacter crescentus is an excellent research model for use in the investigation of DNA replication and cell-cycle regulation due to its unique asymmetric cell division pattern with restricted replication initiation; however, little is known about the specific features of C. crescentus DnaN (CcDnaN). Here, we report a significant divergence in the association of CcDnaN with proteins based on docking analysis and crystal structures that show that the β-binding motifs of its protein partners bind a novel pocket instead of the canonical site. Pull-down and isothermal titration calorimetry results revealed that mutations within the novel pocket disrupt protein-CcDnaN interactions. It was also shown by replication and regulatory inactivation of DnaA assays that mediation of protein interaction by the novel pocket is closely related to the performance of CcDnaN during replication and the DnaN-mediated regulation process. Moreover, assessments of clamp competition showed that DNA does not compete with protein partners when binding to the novel pocket. Overall, our structural and biochemical analyses provide strong evidence that CcDnaN employs a noncanonical protein association pattern.

Aims To evaluate the therapeutic benefits of Hesperidin (Hes) using an imiquimod (IMQ)-induced psoriasis-like mouse model and human immortalized keratinocytes (HaCaT) cells stimulated with lipopolysaccharide (LPS). Methods Mice were treated with IMQ and orally administered Hes (125–500 mg/kg/day), methotrexate (MTX) 1 mg/kg/day or distilled water. HaCaT cells were stimulated with LPS (1 μg/mL) and relevant indices were measured after administration with different concentrations of Hes (5–20 μg/mL) for 24 h. Inflammatory skin lesions in IMQ mice were evaluated using the psoriasis area severity index (PASI) and pathological staining. Proteins in the IRS-1/ERK1/2 pathway and inflammatory factors were assessed using western blotting or quantitative real-time PCR. In addition, factors related to IRS-1 secretion levels were assessed by enzyme-linked immunosorbent assays. Extracellular flux (XF) analysis was used to assess cellular metabolic levels. Key findings Hes significantly improved psoriasis-like skin lesions of IMQ-treated mice and inhibited LPS-induced HaCaT cell proliferation. In addition, Hes remarkably decreased PASI scores, reduced epidermal thickness, decreased proliferation and differentiation of epidermal cells, inhibited mRNA expression of inflammatory factors, reduced local skin lesions and serum insulin and glucose levels. Furthermore, Hes modulated the secretion levels of serum Leptin, Adiponectin and Resistin, and inhibited the activation of the IRS-1/ERK1/2 signaling pathway and regulated HaCaT cells metabolism. Significance This study demonstrated that Hes administration could have significant therapeutic value for the prevention and clinical treatment of psoriasis.

Gastric cancer is a common digestive tract tumor and the second most prevalent cancer. The prognosis of advanced gastric cancer is poor. Conversion therapy can reduce tumor burden, downgrade tumor, and increase the possibility of complete resection, thus prolonging the survival time of patients with gastric cancer. In conversion therapy, chemotherapy and targeted therapy are the main methods of medical treatment, which can control tumor growth and recurrence. As an antiangiogenic targeted drug, apatinib is widely used in the third-line treatment of advanced gastric cancer. Recent studies have shown that it may be of great help in rapid reduction of tumor stage and improvement of prognosis in conversion therapy. This study reported three cases of gastric cancer complicated with multiple abdominal and retroperitoneal lymph node metastases. After receiving apatinib combined with SOX regimen for four cycles, computed tomography showed that the focus and lymph node metastasis were reduced after treatment, and primary tumors were resected. Postoperative pathology result showed that the patients got R0 resection. After radical surgery, the maintenance therapy including apatinib was given. The progression-free survival time was more than 10 months. Apatinib combined with SOX regimen as a conversion therapy for advanced gastric adenocarcinoma increases the possibility of successful surgical resection, which might prolong the survival time of patients.

Using high-performance liquid chromatography (HPLC), a quantitative method was developed to control the quality of ribonucleic acid for injection II (RA-II), which is used as an anti-cancer drug clinically in China. Using nuclease enzyme and under optimal hydrolysis conditions, the unstable RNA was hydrolyzed into stable nucleosides and nucleotides, which were easily detected by HPLC with diode array detection. Furthermore, by analyzing HPLC chromatograms of 10 batches of samples, six common peaks, namely, cytosine nucleotide, uracil nucleotide, adenine nucleotide, guanine nucleotide, guanosine and adenine, were identified by a HLPC coupled with electrospray ionization mass spectrometry. The similarity, the relative retention time and the relative peak area of each common peak, relative to the reference peak, were calculated to obtain the HPLC fingerprints, and their values were within the scope of the provisions. More importantly, all six components were simultaneously determined. Overall, the developed method in this investigation proved to be a useful tool for monitoring the quality of RA-II in terms of their batch variations and the raw material sources.

It has been demonstrated by human genetics that the voltage-gated sodium channel Nav1.7 is currently a promising target for the treatment of pain. In this research, we performed molecular simulation works on a series of classic aryl sulfonamide Nav1.7 inhibitors using three-dimensional quantitative structure-activity relationships (3D-QSAR), molecular docking and molecular dynamics (MD) simulations for the first time to explore the correlation between their structures and activities. The results of the relevant statistical parameters of comparative molecular field analyses (CoMFA) and comparative molecular similarity indices analyses (CoMSIA) had been verified to be reasonable, and the deep relationship between the structures and activities of these inhibitors was obtained by analyzing the contour maps. The generated 3D-QSAR model showed a good predictive ability and provided valuable clues for the rational modification of molecules. The interactions between compounds and proteins were modeled by molecular docking studies. Finally, accuracy of the docking results and stability of the complexes were verified by 100 ns MD simulations. Detailed information on the key residues at the binding site and the types of interactions they participate in involved was obtained. The van der Waals energy contributed the most in the molecular binding process according to the calculation of binding free energy. All research results provided a good basis for further research on novel and effective Nav1.7 inhibitors.

Mingxing Wang, Bo Wu, Sapana N Shah, Peijuan Lu, Qilong Lu McColl-Lockwood Laboratory for Muscular Dystrophy Research, Department of Neurology, Cannon Research Center, Carolinas Medical Center, Charlotte, NC 28203, USA Background: Antisense oligonucleotide (ASO)-mediated exon skipping has been feasible and promising approach for treating Duchenne muscular dystrophy (DMD) in preclinical and clinical trials, but its therapeutic applications remain challenges due to inefficient delivery. Methods: We investigated a few Saponins for their potential to improve delivery performance of an antisense 2′-Omethyl phosphorothioate RNA (2′-OMePS) in muscle cells and in dystrophic mdx mice. This study was carried out by evaluating these Saponins’ toxicity, cellular uptake, transduction efficiency in vitro, and local delivery in vivo for 2′-OMePS, as well as affinity study between Saponin and 2′-OMePS. Results: The results showed that these Saponins, especially Digitonin and Tomatine, enhance the delivery of 2′-OMePS with comparable efficiency to Lipofectamine 2k (LF-2k)-mediated delivery in vitro. Significant performance was further observed in mdx mice, up to 10-fold with the Digitonin as compared to 2′-OMePS alone. Cytotoxicity of the Digitonin and Glycyrrhizin was much lower than LF-2k in vitro and not clearly detected in vivo under the tested concentrations. Conclusion: This study potentiates Saponins as delivery vehicle for 2′-OMePS in vivo for treating DMD or other diseases. Keywords: antisense delivery, 2′-OMePS, exon skipping, saponin, muscular dystrophy

Objectives Indirubin (IR) is a bisindole compound extracted from the leaves of Chinese herb Indigo Naturalis. Indigo Naturalis has been widely used in traditional Chinese medicine to treat inflammatory and autoimmune diseases. Psoriasis is a chronic immune-mediated inflammatory skin disease in which γδ T cells play an important role. This study aims to determine the immunoregulatory effects and the underlying mechanisms of Indirubin in psoriasis-related inflammatory responses. Methods BALB/c mice with imiquimod (IMQ)-induced psoriasis-like dermatitis were treated with saline (Model), 1 mg/kg methotrexate (MTX) that serves as a positive control, or 12.5, 25 and 50 mg/kg Indirubin(IR) intragastrically. Keratinocytes proliferation, inflammatory cells infiltration, the expression of inflammatory cytokines and Jak/Stat pathway-related proteins in the skin lesion were examined. The abundance of γδ T cells in lymph nodes and spleen was determined by flow cytometry. The IL-17 expression and secretion, and the activation of Jak3/Stat3 pathways in in vitro cultured γδ T cell were tested. Results Indirubin ameliorated keratinocyte proliferation, reduced the infiltration of CD3+ T cells, IL-17 A-producing γδ T cells, and CD11b+ neutrophils, inhibited the mRNA expression of Il1, Il6, Il23, Il17a and Il22, and the protein expression of Jak/Stat pathway-related molecules in the skin lesion. Indirubin also reduced the abundance of γδ T cell and CCR6+ γδ T cells (the major IL-17 A producer) in spleen and lymph nodes. In cultured γδ T cells, Indirubin inhibited the mRNA expression of Il17a and Ifng, and the secretion of IL-17 A, while suppressed the activation of Jak3/Stat3 pathways. Conclusion Indirubin alleviates IMQ-induced psoriasis-like dermatitis mainly through reducing the inflammatory responses mediated by IL-17 A-producing γδ T cells involving Jak3/Stat3 activation. Our results highlighted the novel mechanisms by which Indirubin ameliorates psoriasis-related inflammatory responses, supporting its therapeutic potential.

Aims Psoriasis vulgaris is mediated by T and dendritic cells. This study aimed to investigate the effects of acetyl-11-keto-β-boswellic acid (AKBA) on activated dendritic cells (DCs) using an imiquimod (IMQ)-induced psoriasis-like mouse model and murine bone marrow–derived dendritic cells (BMDCs) stimulated with resiquimod (R848) in vitro. Materials and methods The mice were treated with IMQ and intragastrically administered 25–100 mg/kg/day of AKBA, 1 mg/kg/day of methotrexate (MTX), or normal saline. The inflammation of skin lesions in IMQ mice were evaluated by psoriasis area and severity index (PASI) and pathological staining. The related proteins of Toll-like receptor (TLR)7/8 pathways were assessed using Western blotting, and the expression levels of interleukin (IL)-23 and IL-12p40 mRNA using reverse transcription–polymerase chain reaction. The numbers of DCs and marker-positive BMDCs were assessed using flow cytometry and the levels of inflammatory factors using the enzyme-linked immunosorbent assay. Key findings AKBA and MTX obviously improved the psoriasis-like skin lesions of IMQ-treated mice. AKBA also obviously decreased the PASI score, reduced the thickness of epidermis, ameliorated the infiltration of CD3+ and CD11c+ cells in skin lesions, decreased the activation of local DCs, inhibited the mRNA expression and secretion of inflammatory factors IL-12 and IL-23, inhibited the maturation and differentiation of DCs to promote T-cell differentiation, and inhibited the activation of TLR7/8 and IRF signaling pathways. Significance This study implied that AKBA might have an anti-inflammatory effect on psoriasis by inhibiting the maturation and activation of DCs via the TLR8 and IRF signaling pathways.

The establishment of the symbiotic interaction between rhizobia and legumes involves the Nod factor signaling pathway. Nod factor recognition occurs through two plant receptors, NFR1 and NFR5. However, the signal transduction mechanisms downstream of NFR1-NFR5-mediated Nod factor perception remain largely unknown. Here, we report that a small guanosine triphosphatase (GTPase), GmROP9, and a guanine nucleotide exchange factor, GmGEF2, are involved in the soybean-rhizobium symbiosis. We show that GmNFR1α phosphorylates GmGEF2a at its N-terminal S86, which stimulates guanosine diphosphate (GDP)-to-GTP exchange to activate GmROP9 and that the active form of GmROP9 can associate with both GmNFR1α and GmNFR5α. We further show that a scaffold protein, GmRACK1, interacts with active GmROP9 and contributes to root nodule symbiosis. Collectively, our results highlight the symbiotic role of GmROP9-GmRACK1 and support the hypothesis that rhizobial signals promote the formation of a protein complex comprising GmNFR1, GmNFR5, GmROP9, and GmRACK1 for symbiotic signal transduction in soybean.

We investigated a series of Tween 85 modified low molecular weight polyethylenimine (LPEI, 0.8k/1.2k/2.0k)-copolymers (Zs) through simple formulation and covalent conjugation with phosphorodiamidate morpholino oligomer (PMO) for their potential to enhance delivery in vitro and in dystrophic mdx mice. Z polymers significantly enhanced PMO-induced exon-skipping in a GFP reporter-based cell culture system. Application of optimized formulations of Zs with PMO targeted to dystrophin exon 23 demonstrated a significant increase in exon-skipping efficiency in mdx mice. Consistent with our observations in vitro, optimization of molecular size and hydropholic-lipopholic balance (HLB) of polymers are important factors to achieve enhanced PMO delivery in vivo. The best formulation of Zs enhanced PMO delivery with 20- and 6-fold over PMO alone in vitro and in vivo, respectively. Further, chemical conjugation of the polymer and PMO exhibits greater benefit than polymer/PMO simple formulation in PMO delivery efficiency. Observed cytotoxicity of the Zs was lower than Endo-porter and PEI 25k in vitro, and no tissue toxicity was clearly detected with the Zs at the dosage tested. These results indicate the potential of the Zs as effective and safe PMO delivery carriers for treating diseases such as muscular dystrophy., Graphical Abstract

Resists are important multicomponent system require in microelectronic industry for fabrication of integrated circuit (IC) devices. The current high end IC (integrated circuit) technology node in manufacturing is 20 nm and looking toward 16 and 10 nm nodes to fulfil market demand. A series of chemically amplified resists (CARs) based on co and terpolymers of 4-hydroxystyrene (HOST), 2-ethyl-2-adamantyl methacrylate (EAMA) and/or a monomer-bound anionic photoacid generator (PAG) were prepared and characterized. Specifically, Triphenylsulfonium salt 4-(methacryloxy)-2,3,5,6-tetrafluorobenzenesulfonate (F4 PAG) was incorporated into the main chain to get PAG bound polymer microstructure. The aim of the study was to investigate interplay of individual monomers in co and terpolymer systems. This was achieved by understanding the reactivity ratios and monomer sequence distribution in the polymer. Analysis was conducted by using a software PROCOP version2.3. The co/terpolymer compositions were determined by quantitative 1 H NMR spectra. The copolymer reactivity ratio of each monomer was determined with Fineman–Ross, Kelen–Tudos, and nonlinear least-squares curve- fitting methods. It was observed that EAMA is not polymerizable under given reaction conditions and copolymerization of EAMA-PAG was also not possible. HOST-EAMA system yielded reactivity ratios near zero indicating that they form quasi alternating structures. HOST-PAG system yielded copolymers rich in PAG. On the basis of the reactivity ratios determined from the binary copolymers, Alfrey–Goldfinger equations were used to estimate the compositions of the terpolymers. In terpolymer system, where two monomers were non homopolymerizable, both PAG and EAMA competed to add up to the HOST radical. All six reactivity values were less then unity indicating preferred cross propagation of radicals. PAG was the most reactive monomer. Monomer sequence distribution of copolymer and terpolymer was established to get an idea of detailed microstructure of the chain. Copolymers confirmed 84% of dyads formation of each monomer while terpolymer analysis revealed dominance of HOST centered triads. In addition to this, kinetics of terpolymerization reaction was performed to validate the theory. Findings from the study can be used to tailor make CAR synthesis for optimum EUV (extreme ultraviolet) lithography performance.

Paeonol, an active component derived from the traditional Chinese medicine Cortex Moutan, possesses anti-inflammatory, analgesic, antioxidant and anti-allergic properties. Psoriasis is a chronic, recurrent, inflammatory dermatosis accompanied by excessive activation of Toll‑like receptors (TLRs) in dendritic cells (DCs), which are primarily responsible for initiating an immune response. We investigated the effect of paeonol on inflammation in an imiquimod (IMQ)-induced psoriasis-like mouse model and murine bone marrow-derived dendritic cells (BMDCs) stimulated by R848. Mice were intragastrically administered 100 mg/kg (high), 50 mg/kg (medium) and 25 mg/kg (low) paeonol, respectively. We evaluated inflammation of psori-asis‑like lesions based on histological changes, protein levels of myeloid differentiation factor 88 (MyD88) and TLR8 in skin lesions by western blotting, and levels of CD11c+ DCs in skin by immunoassay and in spleens by flow cytometry. Inflammatory cytokines [interleukin (IL)-23, IL-12 and IL-1β] in skin lesions and BMDCs were also assessed by RT-PCR and ELISA. Application of paeonol decreased IMQ-induced keratinocyte proliferation, and infiltration of CD3+ cells, while the treatment ameliorated CD11c+ cells in the spleen and skin, and reduced MyD88 and TLR8 proteins in skin lesions. Paeonol inhibited IMQ-induced mRNA expression of IL-23, but not IL-12 and IL-1β in BMDCs, along with significantly lower levels of DCs expressing MHCⅡ, CD80 and CD86 in vitro. These results indicate that paeonol suppresses the maturation and activation of DCs by decreasing MyD88 and TLR8 proteins in the TLR7/8 signaling pathway which finally alleviates psoriasis‑like skin lesions. The TLR7/8 signaling pathway in DCs provides an important insight into the mechanism of psoriasis, and paeonol may be a potent therapeutic drug for psoriasis.

The glycoprotein from Schisandra chinensis was obtained with alkali extraction and acid precipitation, purified with DEAE Sepharose Fast Flow and Superdex G-75 column. The molecular composition structure and antifatigue activities of glycoprotein were studied. SCGP’s molecular weight was approximately 10 KDa, and it consisted of a carbohydrate component (52.94%) and protein component (47.06%). SCGP comprised mannose, galactoside, rhamnose, glucose, galactose, xylose, arabinose, and fucose, its molar ratio was 2.14 : 1.43 : 1.59 : 8.17 : 8.99 : 3.18 : 18.51 : 1, and it contained 16 kinds of amino acids. SCGP could obviously extend the swimming time in mice by increasing LDH, SOD level, GSH-Px activity, and liver glycogen and decreasing the contents of BUN and MDA. The antioxidant activity of SCGP is a potential mechanism of its antifatigue effect. In vitro antioxidant test showed that SCGP scavenged DPPH and OH radicals in a dose-dependent manner (IC50 was 0.91 mg/ml and 0.72 mg/ml).

A novel oligosaccharide showed that protection against LPS-induced Caco-2 cells was purified from the mycelium of Hericium erinaceus (HE). WEP-1 is mainly composed of neutral monosaccharides with molecular weight of 4,010 Da and of mannose, glucose, and galactose in a molar ratio of 1.2:16.9:1. The structure of WEP-1 includes α-D-Glc (1 → 3) and β-D-Gal (1 → 3) as the backbone with β-D-Glc (1 → 3) as branches attached to the C-4 position and β-D-Man as a terminal residue. The oligosaccharide reduced acetic acid-induced colonic mucosa injury in rats. It also showed significant protection against LPS-induced Caco-2 cells via the TLR4/NF-κB pathway. PRACTICAL APPLICATIONS: In the study, the oligosaccharide from HE has the potential to be developed into functional foods or medicines for the treatment of intestinal diseases. The protection against LPS-induced Caco-2 cells via the TLR4/NF-κB pathway may be a key target for the pharmacological activity of HE.

Here, we focused on exploring the selectivity mechanism against Nav1.7 over Nav1.4 due to different binding modes of two selected inhibitors. By the superposition of Nav1.7 and Nav1.4 proteins, we selected the most homologous chain of Nav1.7 with Nav1.4, defining the active site of Nav1.4-VSD4 based on the aryl sulfonamide binding site of Nav1.7-VSD4. Comparison of the conformations exhibited by Tyr1386 (Nav1.4) and Tyr1537 (Nav1.7) suggested that the steric hindrance caused by Tyr1386 owned primary influence on inhibition selectivity, which was further verified through molecular docking and MD simulation of two representative inhibitors. Our finding would be helpful for discovery of selective Nav1.7 inhibitors over Nav1.4.

Obesity-related renal disease is related to caloric excess promoting deleterious cellular responses. However, a full understanding of the molecular mechanisms involved in progressive kidney disease, as well as a therapeutic strategy, is still absent. Fisetin (FIS), as a natural flavonoid, possesses various bioactivities in a number of disease models. However, its role in obesity-associated kidney injury is still unclear and requires elucidation. In our study, an obesity animal model was established using C57BL/6 mice fed with a normal chow diet (NCD) or high fat diet (HFD) for 16 weeks with or without FIS administration (20, 40 or 80 mg kg-1). Our results indicated that chronic HFD feeding led to a significant body weight gain in mice compared to the normal control group, accompanied by a marked insulin resistance and glucose intolerance, whereas FIS treatment exerted prominently protective effects. In addition, FIS significantly attenuated HFD-induced histological alterations in renal tissue samples. Moreover, FIS treatment down-regulated expression of kidney injury molecule-1 (KIM-1), and up-regulated nephrin and podocin expression levels in the kidneys of HFD-fed mice, improving their renal dysfunction. After HFD feeding, mice treated with FIS exhibited a decrease in phosphorylated IRS1Ser307, and an increase in phosphorylated glycogen synthase kinase 1 (IRS1Tyr608), AKT, forkhead box protein O1 (FOXO1) and glycogen synthase kinase (GSK)-3β. Furthermore, FIS administration markedly restrained the inflammatory response in the kidneys of HFD-challenged mice, as evidenced by the reduced pro-inflammatory cytokines, tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), IL-1β and IL-18, which was attributed to the blockage of nuclear factor κB (NF-κB) signaling. Importantly, FIS-treated obese mice exerted a remarkable decrease in RIP3 expressions in the kidneys compared to obese mice in the absence of FIS, along with an evident reduction in the NOD-like receptor protein 3 (NLRP3), an apoptosis-associated speck-like protein containing a Caspase recruitment domain (ASC) and Caspase-1. The protective effects of FIS against HFD-induced renal injury were verified in vitro using palmitate (PAL)-treated HK2 cells, an immortalized proximal tubule epithelial cell line from the adult human kidney. In summary, our results supported the notion that FIS functions as a promising agent to improve insulin resistance and inflammatory response against metabolic stress-induced renal injury.

A set of triazine-cored cationic amphiphilic polymers (TAPs) composed of low molecular weight (Mw) polyethylenimine (LPEI, B) and amphiphilic Jeffamine (A) were prepared with controllable composition and molecular size, and further characterized for plasmid DNA (pDNA) delivery both in vitro and in vivo. These new polymers condensed pDNA efficiently at a polymer/pDNA weight ratio of 5 with particle sizes below 200 nm. The introduction of Jeffamine in the polymers significantly improved the cellular uptake of pDNA, but without increasing its toxicity compared with the parent LPEI. The best formulation resulted in 6- and 29-fold transfection efficiencies of PEI 25k in vitro and in vivo in mdx mice, respectively. Higher transfection efficiency was achieved with more lipophilic A1/A3-based polymers in vitro, with 1A11B3 and 1A12B3 showing the greatest delivery performance. However, the lipophilicity of the TAPs is less critical in vivo as the less lipophilic A2/A4 constructed TAPs also performed similarly well as the more lipophilic A1/A3 constructed ones. In addition, a synergistic effect of LPEI and Jeffamine via chemical conjugation for the delivery of pDNA was revealed in transfection efficiency. These results indicate that the appropriate positive surface and particle size of polymer/pDNA complex and the composition and hydrophilic-lipophilic balance (HLB) of polymers are crucial for effective delivery, although intricate matching exists between A and B in the TAP composition. Triazine-cored cationic amphiphilic polymers are safe and potentially effective carriers for gene/drug delivery.

Antisense oligonucleotide therapy for Duchenne muscular dystrophy has shown great potential in preclinical and clinical trials, but its therapeutic applications are still limited due to inefficient delivery. In this study, we investigated a few polyquaterniums (PQs) with different size and composition for their potential to improve delivery performance of an antisense phosphorodiamidate morpholino oligomer (PMO) both in vitro and in vivo. The results showed that LuviquatTM series, especially PQ-1 and PQ-3, promoted the exon-skipping efficiency comparable to Endoporter-mediated PMO delivery in vitro. Significant enhancement in skipping dystrophin exon 23 has also been achieved with PQ-3 up to seven-fold when compared to PMO alone in mdx mice. Cytotoxicity of the PQs was lower than Endoporter and PEI 25 K in vitro and muscle damage not clearly detected in vivo under the tested concentrations. These results together demonstrate that the optimization of PQ in molecular size, composition and distribution of positive charges is the key factor to achieve enhanced PMO exon-skipping efficiency. The higher efficiency and lower toxicity endow polyquaternium series as AO delivery enhancing agents for treating muscular dystrophy and other diseases.

The interaction of GluN2B-Containing NMDA Receptor with 18 antagonists were investigated by a combined ligand-based and target-based approach. First, two distinct pharmacophore models were generated for antagonists which cluster in two groups with Catalyst (HipHop module). The pharmacophore of “ifenprodil group” antagonists includes three hydrophobic groups, one H-bond donor and one H-bond acceptor, the pharmacophore of “EVT101 group” antagonists involves one aromatic ring, two hydrophobic groups and one H-bond acceptor. Docking results and pharmacophore model confrontation allow the pharmacodynamic characteristics to be weighted and structural information integrated. Which results in the proposal of two interaction models inside the GluN2B binding cavity for two groups of antagonists. The interaction model of “ifenprodil group” antagonists consists of one hydrophobic group, one H-bond donor, one H-bond acceptor and an aromatic ring, while on the other hand, the interaction model of “EVT101 group” antagonists includes three hydrophobic groups and an aromatic ring.

Glucose-6-phosphate (G6P) is a common alternative carbon source for various bacteria, and its uptake usually relies on the hexose phosphate antiporter UhpT. In the human pathogenic bacterium Staphylococcus aureus, the ability to utilize different nutrients, particularly alternative carbon source uptake in glucose-limiting conditions, is essential for its fitness in the host environment during the infectious process. It has been reported that G6P uptake in S. aureus is regulated by the three-component system HptRSA. When G6P is provided as the only carbon source, HptRSA could sense extracellular G6P and activate uhpT expression to facilitate G6P utilization. However, the regulatory mechanism of HptRSA is still unclear. In this study, we further investigated the HptRSA system in S. aureus. First, we confirmed that HptRSA is necessary for the normal growth of this pathogen in chemically defined medium with G6P supplementation, and we discovered that HptRSA could exclusively sense extracellular G6P compared to the other organophosphates we tested. Next, using isothermal titration calorimetry, we found that HptA could bind to G6P, suggesting that it may be the G6P sensor. After that experiment, using an electrophoresis mobility shift assay, we verified that the response regulator HptR could directly bind to the uhpT promoter and identified a putative binding site from -67 to -96-bp. Subsequently, we created different point mutations in the putative binding site and revealed that the entire 30-bp sequence is essential for HptR regulation. In summary, we unveiled the regulatory mechanism of the HptRSA system in S. aureus, HptA most likely functions as the G6P sensor, and HptR could implement its regulatory function by directly binding to a conserved, approximately 30-bp sequence in the uhpT promoter.

A polysaccharide named EP-1 was found by screening cultured mycelium of Hericium erinaceus, which was extracted and subjected to precipitation with ethanol, hollow-fiber ultrafiltration and ion-exchange chromatography. The polysaccharide has a molecular weight of approximately 3100Da and is composed of glucose, mannose and galactose, thus being a heteroglycan. EP-1 has a backbone of α-d-Glc(1→3) and β-d-Glc(1→3). The β-d-Glc(1→3) and α-d-Gal-(1→3) were regarded as branches attached to the C-4 position. The α-d-Man was regarded as a terminal residue. The anti-CAG activity was evaluated in experimental systems using a cell model for identification. The polysaccharide significantly inhibited the growth of MC cells obtained from human gastric mucosa epithelium (GES-1) cells transformed by MNNG, which were used as a chronic atrophic gastritis cell model. It also interfered with the MC cells by inducing cell cycle arrest. Thus, EP-1 shows potential for the development of new functional foods and drugs.

Mingxing Wang, Bo Wu, Jason D Tucker, Peijuan Lu, Qilong Lu Department of Neurology, McColl-Lockwood Laboratory forMuscular Dystrophy Research, Cannon Research Center, Carolinas Medical Center, Charlotte, NC, USA Abstract: In this study, we investigated a series of cationic polyelectrolytes (PEs) with different size and composition for their potential to improve delivery of an antisense phosphorodiamidate morpholino oligomer (PMO) both in vitro and in vivo. The results showed that the poly(diallyldimethylammonium chloride) (PDDAC) polymer series, especially PE-3 and PE-4, improves the delivery efficiency of PMO, comparable with Endoporter-mediated PMO delivery in vitro. The enhanced PMO delivery and targeting to dystrophin exon 23 was further observed in mdx mice, up to fourfold with the PE-4, compared with PMO alone. The cytotoxicity of the PEs was lower than that of Endoporter and polyethylenimine25,000 Da in vitro, and was not clearly detected in muscle in vivo under the tested concentrations. Together, these results demonstrate that optimization of PE molecular size, composition, and distribution of cationic charge are key factors to achieve enhanced PMO exon-skipping efficiency. The increased efficiency and lower toxicity show this PDDAC series to be capable gene/antisense oligonucleotide delivery-enhancing agents for treating muscular dystrophy and other diseases. Keywords: cationic polyelectrolytes, antisense delivery, exon-skipping, PMO, muscular dystrophy

Antisense oligonucleotide (AON) therapy for Duchenne muscular dystrophy has drawn great attention in preclinical and clinical trials, but its therapeutic applications are still limited due to inefficient delivery. In this study, we investigated a few saponins for their potential to improve delivery performance of an antisense phosphorodiamidate morpholino oligomer (PMO) both in vitro and in vivo. The results showed that these saponins, especially digitonin and tomatine, improve the delivery efficiency of PMO comparable to Endo-Porter-mediated PMO delivery in vitro. The significant enhancement of PMO targeting to dystrophin exon 23 delivery was further observed in mdx mice up to 7-fold with the digitonin as compared to PMO alone. Cytotoxicity of the digitonin and glycyrrhizin was lower than Endo-Porter in vitro and not clearly detected in vivo under the tested concentrations. These results demonstrate that optimization of saponins in molecular size and composition are key factors to achieve enhanced PMO exon-skipping efficiency. The higher efficiency and lower toxicity endow saponins as gene/AON delivery enhancing agents for treating muscular dystrophy or other diseases.

Difficulties in the determination of polysaccharide content in biological samples has been one of the bottlenecks limiting the development of polysaccharides pharmacokinetics, greatly due to the complicated chemical structure of these compounds. In this study, we established a simple, reproducible and reliable method for the determination of polysaccharide content in biological samples. Polysaccharides were replaced by fluorescein isothiocyanate (FITC) and iodine to generate a complex. The iodine content in the complex was measured by inductively coupled plasma-mass spectrometry (ICP-MS) to indirectly reflect polysaccharide content. We investigated the stability of the complex, carried on methodological validation and proceeded to determine the polysaccharide content in rat blood and organs after oral administration of iodine substituted polysaccharides using ICP-MS. The results showed that the iodine complex was stable in vivo and in vitro. In the bioavailability of polysaccharides, after 1h, the absorption rate of polysaccharides in blood was the highest, reaching 5.84%. After 3h, the bioaccessibility was differently distributed in various organs, reaching 1% on average across organs.

In this paper, the micro-structure of amphiphilic copolymer Polylactic acid-Polyethylene glycol-Folate (PLA-PEG-FA) was studied firstly by a differential scanning calorimetry (DSC). During the process of nanoparticles (NPs) preparation, we found good inter-structure consistency of polymer was the precondition for forming into stable NPs, and those with micro-phase separation structure were prepared of NPs within limits. Hemolytic test and CCK-8 assay results demonstrated the biotoxicity of both NPs and whose leaching liquor was far below related toxicity standards. Two kinds of cell, human breast cancer cell line (MCF-7) and human umbilical vein endothelial cells (EC), showed different manners in test of NPs size-cell proliferation relationship, respectively. Monitored by a nuclear magnetic resonance (NMR) and a gel permeation chromatography (GPC), the degradation behavior of NPs in aqueous solution indicated amide bond break more difficultly than ester bond, and FA classic proton peak disappeared in the third week, meanwhile lactic acid (LA) unit number became 25% of the initial. Finally the NPs was completely degraded in the eighth week. In the whole process, NPs underwent a change from compact to loose state. We hope these results will benefit to improve design of drug delivery system in nanomedicine, which could offer the selection rule for amphiphilic polymer NPs on material and size.

Hyperbranched poly(ester amine)s (PEAs) based on tris[2-(acryloyloxy)ethyl]isocyanurate (TAEI) cross-linked low-molecular-weight polyethylenimine (Mw: 0.8k/1.2k/2.0k) have been evaluated for delivering antisense phosphorodiamidate morpholino oligomer (PMO) in vitro and in vivo in the dystrophic mdx mouse. The results show that the PEAs constructed with polyethylenimine (PEI) 2.0k (C series) improved PMO delivery more efficiently than those constructed with PEI 0.8k (A series) or 1.2k (B series) in a GFP reporter-based C2C12 mouse myoblast culture system. The highest efficiency of exon-skipping in vitro with the PMO oligonucleotide targeting human dystrophin exon 50 was obtained when the PEA C12 [TAEI-PEI 2.0k (1:2)] was used. Nearly all of the PEAs improved dystrophin expression in mdx mice by local injection with a 2–4-fold increase when compared with PMO alone. Improved transfection efficiency and lower toxicity indicate the potential of the biodegradable PEA polymers as safe and efficient PMO delivery vectors for in vivo applications.

To study the impacts of emission controls on aerosol physical and chemical properties, real-time measurements of size-resolved aerosol number concentration and chemical composition were conducted in urban Beijing during the 2014 Asia-Pacific Economic Cooperation (APEC) summit, in a period that a series of measures, for example shutting down or halting production from factories and power plants, and restricting the number of vehicles on the roads were implemented in Beijing and surrounding regions. Significantly, reductions in particle mass concentration (55% for PM2.5 and 48% for PM10) were observed during the APEC summit. A clear decrease in secondary inorganic aerosols (SIA), such as sulphate, nitrate and ammonium, was found during APEC, with the reduction ranged from 65.7 to 72.2% for PM1, in which sulphate showed the largest decrease compared with periods before APEC. As a comparison, organics showed a much smaller decrease of 44.3% for PM1 during APEC. These changes were mainly caused by large reductions in accumulation mode particles, which decreased by 36% compared with 19% for Aitken mode particles. The results from the positive matrix factorization (PMF) of particle number concentration indicate that regionally transported aerosols showed significant decreases (70%), similar to those of SIA during APEC, whereas primary factors from traffic and local combustion sources presented much smaller decreases, with the reduction ranged from 4 to 40%. The elevated contributions of these sources indicated the presence of strong local source emissions. The changes in particle chemical composition, size distribution and sources during the evolution of pollution episodes with and without emission controls are further illustrated. Our results highlight the importance of regional atmospheric transport in the formation of severe pollution episodes in Beijing, indicating that reducing the precursors of secondary aerosol over regional scales represent the key steps to reduce the urban particulate pollution. However, stricter emission controls on local source emissions are needed to further mitigate air pollution in Beijing.

A series of poly (ester amines) (PEAs) constructed from low molecular weight polyethyleneimine (LPEI, Mw: 0.8k, 1.2k Da) and Pluronic (different molecular weight (Mw) and hydrophilic-lipophilic-balance (HLB)) components were synthesized, and evaluated in vitro and in vivo as gene delivery carriers. Most PEA polymers were able to bind and condense plasmid DNA effectively into particles of approximately 150 nm in solution at the polymer/DNA ratio of 5 and above. Transfection efficiency of the PEA polymers depends on particle size of the polymer/DNA complex, molecular weight and HLB of the Pluronics and the size of PEI within PEA composition, as well as the cell type. Significant improvement in gene delivery efficacy was achieved with PEA01/04/05 composed of Pluronic size (Mw: 3000-5000 Da), and HLB (12-18) in CHO, C2C12 and HSkM cell lines; and the effective transfection was reflected with PEA 01/04/07 composed of Pluronics with size (2000-5000 Da) and HLB (12-23) in mdx mice. The best formulation for pDNA delivery was obtained with PEA 01 producing transgene expression efficiency 5, 19-folds of that of PEI 25k in vitro and in vivo, respectively. These results potent some of these PEA polymers as attractive vehicles for gene or oligonucleotide delivery.

Population studies have consistently shown a highly inverse correlation between plasma concentration of high-density lipoprotein and the risk of atherosclerotic cardiovascular disease in humans. High-density lipoprotein (HDL) as a therapeutic target is an intense area of ongoing investigation. Aiming to solve the shortcomings of native HDL application, we prepared recombinant human HDL (rhHDL) that contains a similar composition and has similar functions with native HDL. Six kinds of recombinant human apolipoproteins (rhapo)-rhapoA-I, rhapoA-II, rhapoA-IV, rhapoC-I, rhapoC-II, and rhapoE-were expressed in Pichia pastoris and purified with chromatography. By the facilitation of cholate, six kinds of rhapo penetrated among the phosphatidylcholine acyl chains. After purification by density-gradient centrifugation, rhHDL was acquired. Based on morphological observation, we confirmed that the micellar complexes of rhapo with phosphatidylcholine and cholesterol were prepared. We carried on comparative studies in vitro and in vivo between native HDL and rhHDL. Cellular cholesterol efflux assays showed that rhHDL could promote the efflux of excess cholesterol from macrophages. Furthermore, rhHDL has similar effects with native HDL on the blood lipid metabolism in hyperlipidemic mice. In conclusion, rhHDL has similar effects on antiatherosclerosis with native HDL through reverse cholesterol transport, antioxidative, and antithrombotic properties. It could be used as a therapeutic HDL-replacement agent.

A new series of fluorinated anionic photoacid generators (PAGs) (F4-MBS-TPS, F4VBzBS-TPS, F4-IBBS-TPS, CF3 MBS-TPS, MTFBS-TPS and VBzTFBS-TPS), and corresponding PAG bound polymeric resists (HS-EA-PAG) based on hydroxystyrene (HOST) and 2-ethyl-2-adamantyl methacrylate (EA), (GB-EA-PAG) based on γ-butyrolactone methacrylate (GBLMA) and 2-ethyl-2-adamantyl methacrylate (EA) were prepared and characterized. The acid generating efficiency of PAG bound polymers (HS-EA-PAG) series was in the range of 54–81%, which agrees well with the electron withdrawing effect of the substituents. With regard to the referenced F4-PAG bound polymer with 68% acid generating efficiency, and our previously reported EUVL results of F4-MBS-TPS bound polymer photoresists, these new PAG bound polymers should be effective resists for 193 nm or EUV lithography.

A series of poly(esteramine)s (PEAs) constructed from low molecular weight polyethyleneimine (LPEI) and Pluronic were evaluated for the delivery of antisense oligonuclotides (AOs), 2′-O-methyl phosphorothioate RNA (2′-OMePS) and phosphorodiamidate morpholino oligomer (PMO) in cell culture and dystrophic mdx mice. Improved exon-skipping efficiency of both 2′-OMePS and PMO was observed in the C2C12E50 cell line with all PEA polymers compared with PEI 25k or LF-2k. The degree of efficiency was found in the order of PEA 01, PEA 04 > PEA 05 > others. The in vivo study in mdx mice demonstrated enhanced exon-skipping of 2′-OMePS with the order of PEA 06 > PEA 04, PEA 07 > PEA 03 > PEA 01 > others, and much higher than PEI 25k formulated 2′-OMePS. Exon-skipping efficiency of PMO in formulation with the PEAs were significantly enhanced in the order of PEA 02 > PEA 10 > PEA 01, PEA 03 > PEA 05, PEA 07, PEA 08 > others, with PEA 02 reaching fourfold of Endo-porter formulated PMO. PEAs improve PMO delivery more effectively than 2′-OMePS delivery in vivo, and the systemic delivery evaluation further highlight the efficiency of PEA for PMO delivery in all skeletal muscle. The results suggest that the flexibility of PEA polymers could be explored for delivery of different AO chemistries, especially for antisense therapy.

Three novel polymeric anionic and one cationic photoacid generators(PAGs), as well as corresponding PAG bound polymers, were prepared in moderate to good yield and characterized. As for the lithographic properties, the anionic PAG bound polymers showed improved resolution and photospeed than the cationic PAG bound polymer. Especially, the fluorine PAG bound polymer resist HS-EA-PAG(c) showed high photospeed and improved resolution than other PAG bound polymer. Therefore, it should be potential for these new PAG and PAG bounder polymers to be applied to EUV lithography.

The UV–vis absorption and fluorescence spectra, solubility, and application as emitters in single-layer EL devices of novel perylene-3,4:9,10-tetracarboxidiimide-centred dendrimers (Gn-Perys) were examined. Gn-Perys showed absorption maxima (λmax) at 459, 491, and 529 nm in chloroform. The emission maxima (λem) were observed at 537, 576, and 621 nm. The fluorescence intensities of G1-, G2-, and G3-Perys were ca. one-twentieth compared with that of G0-Pery. However, the concentration-quenching of Gn-Perys (n = 1, 2, 3) was improved. The solubility of Gn-Perys increased at higher generation. The electroluminescence (EL) maxima of Gn-Perys in single-layer EL devices containing PVK and PBD as host materials were observed at around 430 and 600 nm, there being white emission. No remarkable difference in the EL characteristics was observed among Gn-Perys.

Two new monomeric anionic PAGs, and their corresponding polymers, were prepared and characterized. The acid generating efficiency of PAG bound polymers is in the order: NO2 PAG ≈ F4 PAG > CF3 PAG, which agrees well with the electron withdrawing power of the substituents. The NO2 PAG bound polymer showed better imaging resolution, than the CF3 PAG bound polymer. Based on the acid generating efficiency, and our previous results on F4 PAG bound polymer for EUV photoresists, these two new PAGs and PAG bound polymers have the potential for imaging via EUV lithography.

Chemically amplified systems, which provide improved sensitivity, are composed of a polymer matrix resin and small molecule photoacid generating units. The inherent discrepancies of the physical and chemical properties of these components can lead to phase separation thereby reducing resolution and sensitivity. Incorporating the PAG in the polymer main chain provides a unified system, which alleviates these problems. Furthermore, PAG incorporated in the chain may improve outgassing and acid diffusion control. Incorporated in the polymer chain was phenylmethacrylate dimethylsufonium triflate photoacid generator (Tf-PAG), which upon exposure generates triflic acid, providing the level of sensitivity required by chemical amplification (CA) systems. 2-Ethyl-2-adamantyl methacrylate (EAMA), and 3-hydroxy-1-adamantyl methacrylate (HAMA) served as the protecting group. (-Butyrolactone methacrylate (GBLMA) provided necessary substrate adhesion. PAGs bound in the main chain resist were exposed with the ASML PAS 5500/950B 193 nm Step and Scan System, with 0.63 NA. Resolution of 110nm line space (220 pitch) and 80 nm isolated lines, with photospeed of 8.2 mJ/cm2 and 2.5 mJ/cm2 respectively, were obtained for these PAG in the chain resists.

1,2,3,4-Tetrafluoro-9,10-anthraquinones reacted with a molar amount of bifunctional nucleophiles, such as 2-aminophenol, catechol, 2-aminobenzenethiol, ethylenediamine, and 1,4-butanediamine to produce the 1,2-cyclized products, which further reacted with another molar amount of bifunctional nucleophiles to afford the dicyclized derivatives.

A range of novel phthalocyanine structures containing polyfluoroalkyloxy groups at the axial sites have been successfully synthesised. They were tested for solubility in a number of solvents. The polyfluoroalkyloxy groups were found to impart higher solubility than the corresponding polyfluoroalkanoyloxy and alkoxy derivatives. The highest solubility was found in ether.

Branched poly(ester amine)s (PEAs) based on tris[2-(acryloyloxy)ethyl]isocyanurate (TAEI) cross-linked low molecular weight polyethylenimine (LPEI, Mw: 0.8k/1.2k/2.0k) polymers have been evaluated for their ability to deliver antisense 2′-O-methyl phosphorothioate RNA (2′-OMePS) through in vitro and in dystrophic mdx mice. Almost all PEAs improved exon-skipping of 2′-OMePS both in vitro and in vivo. Especially the PEAs composed of PEI 2.0k (C series) enhanced 2′-OMePS delivery more effectively than PEI 0.8k or 1.2k-based series using a GFP reporter-based C2C12 myoblast culture system and in mdx mice. The highest efficiency of targeted exon-skipping of 2′-OMePS was obtained by PEA C14 [TAEI-PEI 2.0k (1:4)] with up to 6 fold increase compared with 2′-OMePS only in mdx mice via intramuscular injection without increased muscle damage. Enhanced exon-skipping and lower toxicity highlight the potential of this series biodegradable PEAs as safe and efficient 2′-OMePS delivery vectors for treating diseases such as muscular dystrophy with splicing modulation.

Hericium erinaceus (HE) has been used both as a traditional Chinese medicine and home remedy for treatment of gastric and duodenal ulcers and gastritis. EP-1, a purified polysaccharide isolated from HE mycelium, has recently been identified as the active component responsible for HE anti-gastritis activity. Because oxidative stress has been implicated as a pathogenic cause of gastritis and gastric ulcers, EP-1 antioxidant properties were systematically examined in vitro using the human gastric mucosal epithelial cell line, GES-1. Results showed that EP-1 possessed higher oxygen radical absorbance capacity (ORAC) and 2–3 times higher ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide and hydroxyl radicals than a hot water extract of commercially available HE fruiting body. A crude mycelial polysaccharide (CMPS) extract of HE, from which EP-1 was purified, showed slightly stronger radical scavenging activity and ORAC than EP-1, with the exception of DPPH-scavenging activity. Antioxidant activities of these extracts were further studied using hydrogen peroxide (H2O2)-abused GES-1 cells; EP-1 dose-dependently preserved cell viability of abused cells as assessed via MTT assay. Moreover, FACS analysis revealed that EP-1 prevented H2O2-induced apoptotic cell death by inhibiting activation of apoptotic cellular signals within mitochondria-dependent apoptotic pathways. CMPS also prevented H2O2-induced oxidative stress, but to a lesser degree than did EP-1, even though CMPS exhibited comparable or stronger in vitro antioxidant activity than did EP-1.

A global two-dimensional chemistry model is used to study the long-term trends of CH4, CO, and OH from pre-industrial times to 2020 with given emission scenarios according to the increase of world population. The calculated global-averaged concentration of CO is 27 ppbv before 1840, the concentration of CO is 76 ppbv in 1991, and is estimated to be 105 ppbv in 2020. From 1840 to 1991, the concentration of OH changed from 7.17×105 to 5.79×105 molecules/cm3, i.e., decreased by 19%. The long-term trends of CH4 derived from the model are in good agreement with observation results. The annual increase of CH4 during 1983–1991 is 12.1–13.3 ppbv by this model and 11.1–11.6 ppbv by observation. The calculated growth rate of CO in 1980s is 1.03–1.06%/yr i.e., 6.9–7.9 ppbv/yr. The model is used to investigate why the CO concentration decreased at the beginning of 1990s. We find that the decrease of CO emissions and depletion of stratospheric ozone are the best explanation, which account for 70% and 30% of the decrease of CO concentration, respectively. The model results also show that possible reduction of CH4 emission has little influence on the change of CO concentrations though the reduction of CO emission can counteract the growth of CH4 significantly.

The concentration of atmospheric methane in Beijing is still increasing, although its annual trend has significantly decreased from 2.0% yr −1 in 1985–1989 to 0.5% yr −1 in 1990–1997. The seasonal variability of methane concentration apparently appeared in a double-peak pattern with one peak in winter and the other in summer. It is known that the seasonal inter-annual variations of atmospheric methane in Beijing are different from year to year. From 1986 to 1997, the concentrations of atmospheric methane increased by 184 ppbv, in which about 37% was due to its increase in winter and 21% in summer. After 1993, the trends of methane concentration in summer, which are mainly due to emission from biogenic sources, are negative, while their trends in winter, which are mainly due to emission from non-biogenic sources, are positive with a value of about 25 ppbv yr −1 . As a result, the seasonal inter-annual trends from 1993 to 1997 were mainly due to the increase of methane emission from non-biogenic sources in winter. It implies, therefore, that in Beijing the biogenic sources have been decreasing but the non-biogenic ones, such as fossil fuel combustion, have increased.

Based on the in situ measurement of soil moisture and nitrous oxide (N2O) emission from a rice–wheat rotation ecosystem of southeast China and on the simulated experiments in laboratory, the impact of soil moisture on N2O emission is investigated. By analyzing the experimental data in detail, some results could be outlined as follows: (a) It is soil moisture and temperature instead of N fertilization that determines the seasonal variation pattern of N2O emission from the rice-based crop rotation ecosystem of southeast China. (b) Soil moisture is the most sensitive factor to regulate N2O emission from croplands. (c) Explosive emission of N2O from the rice-based agro-ecosystem was found to happen at the soil moisture within (110±5)% soil water holding capacity or field capacity (SWHC) or (99±9)% water-filled pore space (WFPS). When soil moisture of the rice–wheat fields is less than 105% SWHC, the N2O emission was observed to increase exponentially vs. soil moisture. In contrast, N2O emission was found to decrease reciprocally vs. soil moisture more than 115% SWHC. (d) The response of the N2O emission rate from soils in fields to variations of soil moisture may be well described with a general empirical equation. For x⩽C0% SWHC, F=A e −B(x−C 0 ) 2 +D e Ex . For x⩾C0% SWHC, F=A e −B(x−C 0 ) 2 + e G x −H . The equation to describe the relationship between soil moisture and N2O emission rates from incubated soil is different from that for fitting data observed in fields. Reasons for the difference still remains uncertain.

On the basis of meteorological fields provided by a mesoscale nonhydrostatic model (MM5), a three-dimensional regional chemical model (RADM) is applied to China to address the photochemical reaction mechanisms of surface ozone, with respect to ozone precursors such as NOx and NMHC. The behaviors of surface ozone in the Tibetan Plateau are much different than that in eastern China. Physical factors rather than photochemical factors may play important roles in determining surface ozone. Several model experiments are conducted to focus on these questions. Some results can be obtained as follows: (1) In polluted areas, the surface ozone is primarily dependent on its precursors (NOx, NMHC, etc.) and has highly nonlinear sensitivities to their perturbation. However, in clean areas photochemistry is not the controlling factor. Surface ozone on the Tibetan Plateau is sensitive to ozone perturbation in upper layers, suggesting that ozone above the planetary boundary layer may be a greater influence upon surface ozone. (2) The variations in OH and HO2 caused by perturbations in the precursors are very complicated, and the feedback effect of ozone is very important. There is a nearly linear relationship between percentage differences in HO2 and O3 for NMHC emission reduction, and a similar linear relationship also appears between percentage differences in OH and O3 for NOx emission reduction. (3) During summer midday time, the ratio of O3 to NO2/NO has a constant value.

A two-dimensional global chemistry model is developed to study the distribution and long-term trends of methane. The model contains 34 species and 104 chemical and photochemical reactions. Using the model, the long-term trends of CH4, CO and OH in atmosphere are simulated, comparison between the model and observations shows that the simulation is successful.

A series of small-size polyethylenimine (PEI)-conjugated pluronic polycarbamates (PCMs) have been investigated for the ability to modulate the delivery of 2′-O-methyl phosphorothioate RNA (2′-OMePS) in vitro and in dystrophic mdx mice. The PCMs retain strong binding capacity to negatively charged oligomer as demonstrated by agarose gel retardation assay, with the formation of condensed polymer/oligomer complexes at a wide-range weight ratio from 1:1 to 20:1. The condensed polymer/oligomer complexes form 100–300 nm nanoparticles. Exon-skipping effect of 2′-OMePS was dramatically enhanced with the use of the most effective PCMs in comparison with 2′-OMePS alone in both cell culture and in vivo, respectively. More importantly, the effective PCMs, especially those composed of moderate size (2k–5kDa) and intermediate hydrophilic–lipophilic balance (7–23) of pluronics, enhanced exon-skipping of 2′-OMePS with low toxicity as compared with Lipofectamine-2000 in vitro or PEI 25k in vivo. The variability of individual PCM for delivery of antisense oligomer and plasmid DNA indicate the complexity of interaction between polymer and their cargos. Our data demonstrate the potential of PCMs to mediate delivery of modified antisense oligonucleotides to the muscle for treating muscular dystrophy or other appropriate myodegenerative diseases.