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Wang K.,Zhejiang University | Zhang J.,Zhejiang University | Ping S.,Zhejiang University | Ma Q.,Zhejiang Chinese Medical University | And 5 more authors.
Journal of Ethnopharmacology | Year: 2014

Ethnopharmacological relevance Propolis is used widely in a number of cultures as a folk medicine and is gaining wider recognition for its potential therapeutic use, due to its wide range of biological properties and pharmacological activities, especially its anti-inflammatory effects. Despite an increasing number of studies focused on the biological activities of propolis together with its botanical sources, studies on Chinese propolis are insufficient. This study was designed to investigate the anti-inflammatory properties of ethanol extracts from Chinese propolis (EECP) and poplar buds (EEPB) from Populus×canadensis Moench (Salicaceae family). Materials and methods Phytochemical analysis of EECP and EEPB was performed via total phenolic and flavonoid content measurements followed by high-performance liquid chromatography (HPLC) analysis. DPPH and ABTS free-radical scavenging methods were used to evaluate their anti-oxidant properties. The anti-inflammatory effects of EECP and EEPB were investigated in vitro by evaluating their modulating effects on the key inflammatory cytokines and mediators in LPS/IFN-γ co-stimulated RAW 264.7 cells and by measuring nuclear factor (NF)-κB activation in TNF-α or IL-1β stimulation HEK 293 cells using reporter gene assays. Their effects on acute inflammatory symptoms (LPS-induced endotoxemia and acute pulmonary damage) were also examined in mice. Results EECP and EEPB exhibited strong free-radical scavenging activity and significant in vitro anti-inflammatory effects by modulating key inflammatory mediators of mRNA transcription, inhibiting the production of specific inflammatory cytokines, and blocking the activation of nuclear factor (NF)-κB. The administration of EECP and EEPB (25 and 100 mg/kg) provided significant protective effects by attenuating lung histopathological changes and suppressing the secretion of LPS-stimulated inflammatory cytokines, such as interleukin-6 (IL-6), IL-10, MCP-1, TNF-α and IL-12p70 production in endotoxemic mice. Conclusions The results presented here reveal the potent anti-inflammatory properties of Chinese propolis and poplar buds, and provide biological information for developing suitable substitute(s) for propolis in the prevention and treatment of inflammatory diseases. © 2014 Elsevier Ireland Ltd. All rights reserved.


Xi L.,Zhejiang University | Shou D.,Zhejiang Academy of Traditional Chinese Medicine | Wang F.,Huzhou Institute for Food and Drug Control
Journal of Electroanalytical Chemistry | Year: 2015

A new method for the fabrication of monodispersed Cu nanoparticles on electrode surface was explored, and a novel electrochemical sensor for the sensitive detection of hydrogen peroxide (H2O2) was constructed by assembling copper nanoparticles (Cu-NPs) on the precursor film poly-p-aminobenzene sulfonic acid (poly-ABSA) electropolymerized on glassy carbon electrode (GCE). The assembly process consisted of two steps: the ion-exchange and coordination of Cu2+ with the sulfonic groups contained in poly-ABSA film, and the subsequent in-situ reduction of Cu2+ ions to Cu-NPs by electrochemical method. The results of scanning electron microscopy (SEM) showed that Cu-NPs were highly monodispersed in whole poly-ABSA, which was strongly attached to the electrode surface. The modified electrode Cu-NPs/poly-ABSA/GCE was also characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS), and exhibited excellent electrocatalytic activity toward the reduction of H2O2 with high stability. Amperometry was used for the determination of H2O2 at a more positive potential of -0.2 V versus Ag/AgCl in pH 7.0 phosphate buffer solution, with a linear range from 0.2 μM to 2.32 mM and a detection limit of 0.1 μM. The Cu-NPs/poly-ABSA/GCE allowed sensitive, stable and fast amperometric sensing of H2O2, and this was promising for the development of nonenzymatic sensors. The analysis of medical disinfection solution real samples was performed using the proposed method, and the obtained results were satisfactory. © 2015 Published by Elsevier B.V.


Hu Y.,Zhejiang University | Hu Y.,Zhejiang Pharmaceutical College | Xu B.,Zhejiang Pharmaceutical College | Ji Q.,Zhejiang Pharmaceutical College | And 6 more authors.
Biomaterials | Year: 2014

Polyethylenimine (PEI) is widely applied in non-viral gene delivery vectors. PEI with high molecular weight is highly effective in gene transfection but is high cytotoxic. Conversely, PEI with low molecular weight displays lower cytotoxicity but less delivering efficiency. To overcome this issue, a novel copolymer with mannosylated, a cell-penetrating peptide (CPP), grafting into PEI with molecular weight of 1800 (Man-PEI1800-CPP) were prepared in this study to target antigen-presenting cells (APCs) with mannose receptors and enhance transfection efficiency with grafting CPP. The copolymer was characterized by 1H NMR and FTIR. Spherical nanoparticles were formed with diameters of about 80-250nm by mixing the copolymer and DNA at various charge ratios of copolymer/DNA(N/P). Gel retardation assays indicated that Man-PEI1800-CPP polymers efficiently condensed DNA at low N/P ratios. Cytotoxicity studies showed that Man-PEI1800-CPP/DNA complexes maintained in a high percentage of cell viability compared to the PEI with molecular weight of 25k (PEI25k). Laser scan confocal microscopy and flow cytometry confirmed that Man-PEI1800-CPP/DNA complexes resulted in higher cell uptake efficiency on DC2.4 cells than on Hela cells line. The transfection efficiency of Man-PEI1800-CPP was significantly higher than that of PEI25k on DC2.4 cells. More importantly, the complexes were mainly distributed in the epidermis and dermis of skin and targeted on splenocytes after percutaneous coating based on microneedles invivo. These results indicated that Man-PEI1800-CPP was a potential APCs targeted of non-virus vector for gene therapy. © 2014 .


Ji L.,Zhejiang University | Yuan Y.,Zhejiang University | Luo L.,Zhejiang University | Chen Z.,Chinese Traditional Medicine Hospital of Zhejiang Province | And 3 more authors.
Steroids | Year: 2012

Michael reaction acceptors (MRAs) are a class of active molecules that are directly or indirectly involved in various cellular processes, including the regulation of many signaling pathways. In this study, the inducible nitric oxide synthase (iNOS) assay was used to demonstrate that the dichloromethane extract of Physalis alkekengi var. franchetii (DCEP) possesses anti-inflammatory activity that might be attributed to the modification of key cysteine residues in IKKβ by the MRAs in DCEP. To isolate these MRAs, glutathione (GSH) was employed, and a simple ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) screening method was developed to investigate the GSH conjugates with potential MRAs. Five physalins, including one new compound isophysalin A (2), together with four known steroidal compounds, physalin A (1), physalin O (3), physalin L (4) and physalin G (5), were isolated to evaluate the GSH conjugating abilities, and it was indicated that compounds 1, 2 and 3, which had a common α,β-unsaturated ketone moiety, exhibited conjugating abilities with GSH and also showed significant nitric oxide (NO) production inhibiting activities. The anti-inflammatory activities of compounds 1, 2 and 3 might be attributed to their targeting multiple cysteine residues on IKKβ; therefore, the alkylation of IKKβ by compound 1 was further studied by micrOTOF-MS. The result showed that six cysteine residues (C 59, C 179, C 299, C 370, C 412, and C 618) were alkylated, which indicated that IKKβ is a potential target for the anti-inflammatory activity of physalin A. © 2011 Elsevier Inc. All rights reserved.


Zhong L.-R.,Zhejiang Chinese Medical University | Chen X.,ZheJiang HuaYi Pharmaceutical Co. | Wei K.-M.,Zhejiang Academy of Traditional Chinese Medicine
Asian Pacific Journal of Cancer Prevention | Year: 2013

Radix Tetrastigma Hemsleyani Flavone (RTHF) is widely used as a traditional herb for its detoxification and anti-inflammation activity. Recently, several studies have shown that RTHF can inhibit growth and induce apoptosis in human cancer cell lines. However, the mechanisms are not completely understood yet. In this study we investigated the potential effects of RTHF on growth and apoptosis in human lung adenocarcinoma A549 cells as well as its mechanisms. A549 cells were treated with RTHF at various concentrations for different times. In vitro the MTT assay showed that RTHF had obvious anti-proliferation effects on A549 cells in a dose- and timedependent manner. Cell morphological changes observed by inverted microscope and Hoechst33258 methods were compared with apoptotic changes observed by fluorescence microscope. Cell apoptosis inspected by flow cytometry showed significant increase in the treatment group over the control group (P<0.01). Expression of apoptosis related Bax/Bcl-2, caspases and MAPK pathway proteins were detected by Western blotting. The results showed that RTHF up-regulated the Bax/Bcl-2 ratio and cle-caspase3/9, cle-PARP expression in a dosedependent manner. Expression of p-p38 increased, p-ERK decreased significantly and that of p-JNK was little changed in the RTHF group when compared with the control group. These results suggest that RTHF might exert anti-growth and apoptosis activity against lung cancer A549 cells through activation of caspases and Bcl-2 family proteins and the MAPK pathway, therefore presenting as a promising therapeutic agent for the treatment of lung cancer.


Yan F.,Zhejiang University | Dai G.,Zhejiang Academy of Traditional Chinese Medicine | Zheng X.,Zhejiang University
Journal of Nutritional Biochemistry | Year: 2016

This study evaluated the capacity of mulberry anthocyanin extract (MAE) on insulin resistance amelioration in HepG2 cells induced by high glucose and palmitic acid and diabetes-related metabolic changes in type 2 diabetic mice. In vitro, MAE alleviated insulin resistance in HepG2 cells and increased glucose consumption, glucose uptake and glycogen content. Enzyme activities of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) were decreased due to PPARγ coactivator 1α (PGC-1α) and forkhead box protein O1 (FOXO1) inhibition. Furthermore, phosphorylation of protein kinase B (AKT) and glycogen synthase kinase-3β (GSK3β) in model cells was recovered after treated with MAE, leading to an up-regulation of glycogen synthase 2 (GYS2), and this effect was blocked by the phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002. In vivo, MAE supplementation (50 and 125 mg/kg body weight per day) markedly decreased fasting blood glucose, serum insulin, leptin, triglyceride and cholesterol levels and increased adiponectin levels in db/db mice. The improvement of related metabolic parameters was in part associated with the impact of MAE on activating AKT and downstream targets in liver, skeletal muscle and adipose tissues. In summary, these findings suggest that MAEs have potential benefits on improving dysfunction in diabetic mice and mitigating insulin resistance in HepG2 cells via activation of PI3K/AKT pathways. © 2016 Elsevier Inc.


Pawlus M.R.,Aurora University | Wang L.,Aurora University | Murakami A.,Aurora University | Dai G.,Zhejiang Academy of Traditional Chinese Medicine | Hu C.-J.,Aurora University
PLoS ONE | Year: 2013

The HIF1- and HIF2-mediated transcriptional responses play critical roles in solid tumor progression. Despite significant similarities, including their binding to promoters of both HIF1 and HIF2 target genes, HIF1 and HIF2 proteins activate unique subsets of target genes under hypoxia. The mechanism for HIF target gene specificity has remained unclear. Using siRNA or inhibitor, we previously reported that STAT3 or USF2 is specifically required for activation of endogenous HIF1 or HIF2 target genes. In this study, using reporter gene assays and chromatin immuno-precipitation, we find that STAT3 or USF2 exhibits specific binding to the promoters of HIF1 or HIF2 target genes respectively even when over-expressed. Functionally, HIF1α interacts with STAT3 to activate HIF1 target gene promoters in a HIF1α HLH/PAS and N-TAD dependent manner while HIF2α interacts with USF2 to activate HIF2 target gene promoters in a HIF2α N-TAD dependent manner. Physically, HIF1α HLH and PAS domains are required for its interaction with STAT3 while both N- and C-TADs of HIF2α are involved in physical interaction with USF2. Importantly, addition of functional USF2 binding sites into a HIF1 target gene promoter increases the basal activity of the promoter as well as its response to HIF2+USF2 activation while replacing HIF binding site with HBS from a HIF2 target gene does not change the specificity of the reporter gene. Importantly, RNA Pol II on HIF1 or HIF2 target genes is primarily associated with HIF1α or HIF2α in a STAT3 or USF2 dependent manner. Thus, we demonstrate here for the first time that HIF target gene specificity is achieved by HIF transcription partners that are required for HIF target gene activation, exhibit specific binding to the promoters of HIF1 or HIF2 target genes and selectively interact with HIF1α or HIF2α protein. © 2013 Pawlus et al.


Ding Y.,Nanjing Medical University | Leng J.,Nanjing Medical University | Fan F.,Zhejiang Cancer Hospital | Xia B.,Hunan University | Xu P.,Zhejiang Academy of Traditional Chinese Medicine
Biochemical Genetics | Year: 2013

Mutations in mitochondrial DNA (mtDNA) are one of the most important causes of hearing loss. Of these, the homoplasmic A1555G and C1494T mutations at the highly conserved decoding site of the 12S rRNA gene are well documented as being associated with either aminoglycoside-induced or nonsyndromic hearing loss in many families worldwide. Moreover, five mutations associated with nonsyndromic hearing loss have been identified in the tRNASer(UCN) gene: A7445G, 7472insC, T7505C, T7510C, and T7511C. Other mtDNA mutations associated with deafness are mainly located in tRNA and protein-coding genes. Failures in mitochondrial tRNA metabolism or protein synthesis were observed from cybrid cells harboring these primary mutations, thereby causing the mitochondrial dysfunctions responsible for deafness. This review article provides a detailed summary of mtDNA mutations that have been reported in deafness and further discusses the molecular mechanisms of these mtDNA mutations in deafness expression. © 2013 Springer Science+Business Media New York.


Zhou N.,Zhejiang University | Li J.,Zhejiang University | Zhang Y.,Zhejiang University | Lu J.,Zhejiang University | And 9 more authors.
Journal of Hepatology | Year: 2015

Background & Aims: Extracorporeal blood purification systems for supportive therapy of liver failure are widely used. We developed a novel blood purification system, named Li's artificial liver system (Li-ALS), which couples low-volume plasma exchange (low-volume PE) with plasma filtration adsorption (PFA). This study aims to evaluate the efficacy of our novel system in pigs with acute liver failure (ALF). Methods: Thirty-two pigs were infused with D-galactosamine (1.3 g/kg) to induce ALF. All animals were equally and randomly divided into four groups: the ALF control group received intensive care, the PFA group underwent five hour plasma recycling filtration and adsorption purification, the low-volume PE group received one hour low-volume PE, and the Li-ALS group underwent one hour low-volume PE, followed by five hour PFA. Intervention was initiated 36 hours after drug administration. The efficacy of each treatment was assessed by survival time and improvement in hematological, biochemical, and immunohistological parameters. Results: Pigs in the Li-ALS group survived longer than those in the other groups (p <0.001, ALF control: 60 ± 2 h; PFA group: 74 ± 2 h; low-volume PE group: 75 ± 2 h; and Li-ALS group: 90 ± 3 h). Liver enzyme, bilirubin, bile acid and blood ammonia levels were decreased significantly after Li-ALS treatment, and increases in inflammatory cytokines were ameliorated. A higher hepatocyte regeneration index was also observed in the Li-ALS group. Conclusion: Our novel Li-ALS could expedite liver regeneration and improve survival time; hence, it could be promising for treating ALF. © 2015 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.


PubMed | Zhejiang Academy of Traditional Chinese Medicine and Zhejiang Cancer Hospital
Type: | Journal: Medical science monitor : international medical journal of experimental and clinical research | Year: 2017

BACKGROUND Wound healing in chronic diabetic mellitus is mainly associated with the management of angiogenesis. The angiogenic mechanism of vascular endothelial growth factor (VEGF) has been widely studied in the context of diabetic ulcers. The aim of this study was to investigate the wound-healing potential of curcumol in streptozotocin-induced diabetic rats. MATERIAL AND METHODS Sixty male SD (Sprague Dawley) rats were purchased and randomly assigned into four groups: a control group and a model group treated with blank ointment, a high-dose curcumol group, and a low-dose curcumol group. The number of animals in each group was 15. Diabetes was induced by an intraperitoneal injection of streptozotocin. Two cutaneous wounds were incised at the dorsal region of all the experimental animals. Wound healing was assessed for all animal groups by observing the rate of wound closure. The expression of VEGF at the wound sites was studied by immunohistochemical staining to evaluate the vascular endothelial cell reaction. VEGF protein and related mRNA levels were analyzed by Western blotting and RT-PCR (reverse transcription-polymerase chain reaction). RESULTS Curcumol treatment significantly increased the rates of wound closure in treated animals, and hence wound healing was drastically enhanced for treatment groups compared to control groups. Histological observations and related mRNA and protein levels showed a higher VEGF expression in the treatment groups. CONCLUSIONS Our analyses clearly suggested that the observed enhancement in wound healing as a result of curcumol administration was attributable to VEGF-mediated angiogenesis.

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