Flavonoids as potential anti-hepatocellular carcinoma agents: recent approaches using HepG2 cell line.

Numerous studies have documented that in cancer therapy flavonoids extracted from traditional Chinese medicine have anti-tumor activity or can enhance efficiency of chemotherapy in combination with chemotherapeutics. Thus, an awareness of flavonoids is needed by physicians and medical researchers. This review provides evidence about anti-hepatocellular carcinoma activity of flavonoids. First, as a common employed in vitro model, profile of HepG2 is shown. Second, the intracellular signaling pathways induced by flavonoids which inhibit the HepG2 cell line are summarized. Third, study situation of anti-HBV/HCV activity of flavonoids is shown. Our review is aimed at providing an understanding of anti-HBV/HCV activity and anti-HCC mechanisms of flavonoids, and an outlook on flavonoids application on cancer therapy.

Long-term infusion of brain-derived neurotrophic factor reduces food intake and body weight via a corticotrophin-releasing hormone pathway in the paraventricular nucleus of the hypothalamus.

Brain-derived neurotrophic factor (BDNF) has been implicated in learning, depression and energy metabolism. However, the neuronal mechanisms underlying the effects of BDNF on energy metabolism remain unclear. The present study aimed to elucidate the neuronal pathways by which BDNF controls feeding behaviour and energy balance. Using an osmotic mini-pump, BDNF or control artificial cerebrospinal fluid was infused i.c.v. at the lateral ventricle or into the paraventricular nucleus of the hypothalamus (PVN) for 12 days. Intracerebroventricular BDNF up-regulated mRNA expression of corticotrophin-releasing hormone (CRH) and urocortin in the PVN. TrkB, the receptor for BDNF, was expressed in the PVN neurones, including those containing CRH. Both i.c.v. and intra-PVN-administered BDNF decreased food intake and body weight. These effects of BDNF on food intake and body weight were counteracted by the co-administration of alpha-helical-CRH, an antagonist for the CRH and urocortin receptors CRH-R1/R2, and partly attenuated by a selective antagonist for CRH-R2 but not CRH-R1. Intracerebroventricular BDNF also decreased the subcutaneous and visceral fat mass, adipocyte size and serum triglyceride levels, which were all attenuated by alpha-helical-CRH. Furthermore, BDNF decreased the respiratory quotient and raised rectal temperature, which were counteracted by alpha-helical-CRH. These results indicate that the CRH-urocortin-CRH-R2 pathway in the PVN and connected areas mediates the long-term effects of BDNF to depress feeding and promote lipolysis.

Dynamic of modernizing traditional Chinese medicine and the standards system for its development.

This article reviewed the process of Traditional Chinese Medicine's modernization on a global scale. This process is motivated by the potential need for traditional medicine as a result of health transitions and increasing drug R&D based on know-how from TCM. The established standards system for modern medicine serves as a basic model yet has limitations in terms of comprehensively evaluating TCM. Spurred by policy committments, research to provide supplements suited to TCM's features and principles is underway. Advanced and interdisciplinary technology and methodology is expected to play an essential role in TCM development.

Japan-China Joint Medical Workshop on Drug Discoveries and Therapeutics 2008: The need of Asian pharmaceutical researchers' cooperation.

The Japan-China Joint Medical Workshop on Drug Discoveries and Therapeutics 2008 (JCMWDDT 2008) was held from September 29 to October 1, 2008 at The University of Tokyo, Tokyo, Japan. JCMWDDT is an international workshop that is mainly organized by Asian editorial members of Drug Discoveries & Therapeutics (http://www.ddtjournal.com/home) for the purpose of promoting research exchanges in the field of drug discovery and therapeutic. This year's JCMWDDT is the second workshop and focused particularly on novel development and technological innovation of anti-influenza agents. The workshop began with an announcement by the Japanese Co-chairperson, Dr. Sekimizu (Department of Microbiology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Japan; Editorin- Chief of Drug Discoveries & Therapeutics, DDT) followed by a speech by the Chinese Co-chairperson, Dr. Wenfang Xu (School of Pharmaceutical Sciences, Shandong University, Shandong, China; Editor in China Office of DDT), with additional speeches by Dr. Norio Matsuki (The University of Tokyo, Japan; Editor of DDT) and Dr. Guanhua Du (Chinese Academy of Medical Science, China; Editor of DDT). Fifty-nine titles were presented in 6 specialized sessions (Research Advances in Drug Discoveries and Therapeutics, Drug Synthesis/Clinical Therapeutics, Medicinal Chemistry/Natural Products, Anti-influenza Drugs, Anti-infection/antiviral Drugs, Biochemistry/Molecular Biology /Pharmacology) and a poster session (Drug Discov Ther 2008; 2, Suppl; available at http://www.ddtjournal.com/Announce/index.htm). An annual outbreak of avian influenza in Asian countries including China and Japan has sparked fears that the virus will mutate and then cause an epidemic in humans. Therefore, Asian researchers need to work together to control this infection. This year's JCMWDDT helped provide an opportunity to reiterate the crucial role of medicinal chemistry in conquering influenza and created an environment for cooperative research in Asian countries. (reported on October 1st, with grateful thanks to all participants) Main program Session I. Research Advances in Drug Discoveries and Therapeutics ● Design, synthesis and preliminary activity assay of influenza virus neuraminidase inhibitors by Wenfang Xu (Shandong University, China) ● Infection disease models with silkworms to evaluate the therapeutic effects of drug candidates by Kazuhisa Sekimizu (The University of Tokyo, Japan) ● Japan's governmental approaches to facilitate drug development process by Makoto Shimoaraiso (Ministry of Foreign Affairs of Japan, Japan) ● Effective detection of the epidermal growth factor receptor mutation by the peptide nucleic acid-locked nucleic acid PCR Clamp by Sakuo Hoshi (The University of Tokyo Hospital, Japan) ● Design and synthesis of p53-MDM2 binding inhibitors by Yongzhou Hu (Zhejiang University, China) Session II. Drug Synthesis/Clinical Therapeutics ● Pharmacogenomics-based clinical studies using a novel fully-automated genotyping system by Setsuo Hasegawa (Sekino Clinical Pharmacology Clinic, Japan) ● Synthesis and biological evaluation of pentacyclic triterpenes as anti-tumor agents by Hongbin Sun (China Pharmaceutical University, China) ● Drug discovery and therapeutics using silkworm as experimental animal by Yasuyuki Ogata (The University of Tokyo, Japan) ● Novel selective estrogen recetpor modulators (SERMs) with unusual structure and biological activities by Haibing Zhou (Wuhan University, China) Session III. Medicinal Chemistry/Natural Products ● Synthesis and properties of isonucleosides incorporated oligonucleotides by Zhenjun Yang (Peking University, China) ● Isolation of antiviral compounds from plant resources using silkworm bioassay by Yutaka Orihara (The University of Tokyo, Japan) ● Synthesis and structural modifcation of tasiamide and the effect of these modifications on in vitro anticancer activity by Yingxia Li (Ocean University of China, China) ● Spirohexalines A and B, novel undecaprenyl pyrophosphate inhibitors produced by Penicillium sp. FKI-3368 by Junji Inokoshi (Kitasato University, Japan) ● Nosokomycins, novel anti-MRSA antibiotics, produced by Streptomyces sp. K04-0144 by OR. Uchida (Kitasato University, Japan) ● In vivo screening for antimicrobial activity of Thai Herbal Medicines using silkworm model by Santad Chanprapaph (Chulalongkorn University, Thailand) ● Novel electrochemical sensor of nitric oxide for screening anti-aging Traditional Chinese Medicine by Zilin Chen (Wuhan University, China) ● Polysacchride from green tea purified by silkworm muscle contraction assay induces innate immunity by increasing the expression of various inflammatory cytokine mRNA in human leukocytes by Saphala Dhital (The University of Tokyo, Japan) Session IV. Anti-influenza Drugs ● Structure-activity relationship of flavonoids as influenza virus neuraminidase inhibitors and their in vitro anti-viral activities by Guanhua Du (Chinese Academy of Medical Sciences and Peking Union Medical College, China) ● Mechanisms and consequences of phagocytosis of influenza virus-infected cells by Yoshinobu Nakanishi (Kanazawa University, Japan) ● Nuclear export inhibitors; a possible target for novel anti-influenza viral drugs by Ken Watanabe (Nagasaki University, Japan) ● Catalytic asymmetric synthesis of oseltamivir phosphate directing toward its stable worldwide supply by Motomu Kanai (The University of Tokyo, Japan) ● Clinical effects of probiotic bifidobacterium in the prevention of influenza virus infections and allergic diseases by Jin-zhong Xiao (Morinaga Milk Industry Co., Ltd., Japan) ● Production of anti-influenza PR8-scFv using a phage display by Normaiza Zamri (Tokai University, Japan) Session V. Anti-infection/Antiviral Drugs ● Emerging infectious diseases and anti-viral drugs: Urgent need to develop effective drugs which cause less resistant virus by Nobuyuki Kobayashi (Nagasaki University, Japan) ● Design, synthesis and antiviral evaluation of novel heterocyclic compounds as HIV-1 NNRTIs by Xinyong Liu (Shandong University, China) ● Antiviral drug screening from microbial products by Eisaku Tsujii (Astellas Pharma Inc., Japan) ● Viral factors that determine the natural course of chronic hepatitis B viral infection by Hiroshi Yotsuyanagi (The University of Tokyo, Japan) ● Effect of andrographolide derivatives having α-glucosidase inhibition, on HBsAg, HBeAg secretion in HepG2 2.2.15 cells by Hongmin Liu (Zhengzhou University, China) ● Current and future antiviral therapy for influenza by Hideki Asanuma (Tokai University, Japan) ● Establishment of an HIV-based pseudotyping system as a safe model for screening inhibitors on bird flu H5N1 entry by Ying Guo (Peking Union Medical Collegee Chinese Academy of Medical Sciences, China) ● Strategy of discovery for novel antibiotics using silkworm infection model by Hiroshi Hamamoto (The University of Tokyo, Japan) ● Potent neuraminidase inhibitors and anti-inflammatory substances from Chaenomeles speciosa by Li Zhang (Chinese Academy of Medical Sciences and Peking Union Medical College, China) ● High-throughput screening assay for hepatitis C virus helicase inhibitors using fluorescence-quenching phenomenon by Hidenori Tani (Waseda University and National Institute of Advanced Industrial Science and Technology, Japan) Session VI. Biochemistry/Molecular Biology/Pharmacology ● A novel conjugate of low-molecular-weight heparin and Cu,Zn-superoxide dismutase: Study on its mechanism in preventing brain reperfusion injury after ischemia in gerbils by Fengshan Wang (Shandong University, China) ● A novel gene fudoh in SCCmec region regulates the colony spreading ability and virulence in Staphylococcus aureus by Chikara Kaito (The University of Tokyo, Japan) ● Water soluble fluorescent boronic acid sensors for tumor cell-surface saccharide by Hao Fang (Shandong Unviersity, China) ● Molecular characterization of the biosynthetic enzyme for the biotechnological production of tetrahydrocannabinol, the active constituent of marijuana by Futoshi Taura (Kyushu University, Japan) ● Galloyl cyclic-imide derivative CH1104I inhibits tumor invasion via suppressing matrix metalloproteinase activity by Xianjun Qu (Shandong University, China) ● Neuroprotection by inhibition of GAPDH-MAO B mediated cell death induced by ethanol by Xiao-Ming Ou (University of Mississippi Medical Center, USA).

The agonist of the protease-activated receptor-1 (PAR) but not PAR3 mimics thrombin-induced vascular endothelial growth factor release in human smooth muscle cells.

Thrombin, a serine protease generated by the activation of the blood coagulation cascade following vessel injury, induces vascular endothelial growth factor-(VEGF) release. However, the molecular mechanism of thrombin-induced VEGF release is largely unknown. Anagonist of protease-activated receptor-i (PARI), SFLL-RNPNDKYEPF, mimicked thrombin-induced VEGF release in human vascular smooth muscle (HVSM) cells, as determined by enzyme-linked immunosorbent assay, reverse transcriptase-polymerase chain reaction, and Northern blotting. In contrast, the agonist of PAR3, TFR- GAP, did not affect VEGF release or expression. SFLL-RNPNDKYEPF, but not TFRGAP, up-regulated [Ca2-]i.Moreover, the calcium ionophone A23187 was found to trigger VEGF release in HVSM cells. Thrombin-inducedVEGF release was blocked by anti-thrombin, heparin, a synthetic thrombin receptor inhibitor E5510, the calcium chelator BAPTA, the protein kinase C inhibitor calphostin C, and the MEK1/2 inhibitor U0126. Thus, our data show that thrombin caused VEGF release via PARI activation in a manner dependent on [Ca2+]i and p44/42 downstream from the receptor activation.

Evaluation of the antioxidant activity of environmental plants: activity of the leaf extracts from seashore plants.

The antioxidant activity of the methanolic extracts of the leaves of 39 plant species was examined. These leaves were collected from the plants growing on subtropical seashores. The activity was evaluated by three kinds of assay methods, which included the DPPH radical scavenging assay, linoleic acid oxidation assay, and oxidative cell death assay. Two extracts from Excoecaria agallocha and Terminalia catappa showed remarkably potent activity in all assay systems. The HPLC analysis of the extracts indicated the presence of the same antioxidant and isolation work for the compound identified ellagic acid. The isolated ellagic acid showed strong antioxidant activity in the assay systems used.

Satiation and masticatory function modulated by brain histamine in rats.

Both the ventromedial hypothalamus (VMH) and the mesencephalic trigeminal sensory nucleus (Me5) are densely innervated by histaminergic neurons. The depletion of neuronal histamine (HA) from the Me5 by the bilateral microinfusion of 448 nmol/rat alpha-fluoromethylhistidine (FMH), a specific suicide inhibitor of histidine decarboxylase, reduced the eating speed and prolonged meal duration, while leaving the meal size unaffected. HA depletion from the VMH increased the size of the meal and prolonged its duration, but not the eating speed. When the HA turnover rate was measured at 15 min after the scheduled feeding following fasting for less than 24 hr, the rate increased in the region including the Me5, but not in the hypothalamus. The turnover rate reached higher levels at 60 min in both regions. Gastric intubation of an isocaloric liquid diet or an equivolume of water with the liquid diet abolished the increase in HA turnover both in the Me5 region and the hypothalamus. The present findings indicate that brain HA thus modulates satiation through both the VMH and masticatory function as well as due to the action of the Me5. The HA function activated by mastication began earlier in the Me5 and later in the hypothalamus due to a signal originating from the oral proprioceptors and initiated by chewing.

A novel human serum lectin with collagen- and fibrinogen-like domains that functions as an opsonin.

Collectins are C-type animal lectins with both collagenous and carbohydrate recognition domains and are involved in the first line host defense against pathogens. We report here a novel Ca(2+)-dependent and GlcNAc-binding lectin consisting of subunits of 35 kDa (P35) with a collagen-like sequence. When P35 is isolated from human serum, it forms a homopolymer by means of intermolecular disulfide bonding, as is the case with collectins. P35 cDNA was cloned from a human liver cDNA library, and the deduced amino acid sequence of 313 residues revealed that the mature form of P35 consists mainly of collagen- and fibrinogen-like domains. The latter contained two potential Ca(2+)-binding sites that may be involved in carbohydrate binding. The overall sequence of P35 was highly homologous to porcine ficolins alpha and beta. Northern blots of various human tissues showed that the major product of the 1.3-kilobase-long P35 transcript is expressed in liver. P35 enhanced phagocytosis of Salmonella typhimurium by neutrophils, suggesting an opsonic effect via the collagen region. P35 was found to bind to GlcNAc-conjugated bovine serum albumin, a neoglycoprotein, as well as to neoglycolipids containing complex-type oligosaccharides derived from glycoproteins, suggesting that P35 recognizes GlcNAc residues such as those found in microbial glycoconjugates and complex-type oligosaccharides. Therefore, P35 represents a new type of GlcNAc-binding lectin with structural and functional similarities to collectins involved in innate immunity.

Expression of human platelet-activating factor receptor gene in EoL-1 cells following butyrate-induced differentiation.

Platelet-activating factor (PAF) is a potent lipid mediator of allergic inflammation through its interaction with eosinophils. Expression of the PAF receptor is modulated by many agents, including those responsible for cell differentiation. We report here that differentiation of a human eosinophilic leukaemia cell line, EoL-1, by sodium n-butyrate is associated with induction of PAF receptor gene expression, as indicated by: PAF receptor mRNA accumulation; increases in the binding of [3H]WEB 2086, a PAF antagonist; analysis of cell-surface expression of PAF receptor protein using a monoclonal anti-(PAF receptor) antibody; and augmentation of PAF-induced increase in the intracellular concentration of calcium. Using cDNA cloning, the receptor expressed in EoL-1 cells was identified as 'Transcript 1', one of two transcripts which was previously reported from human genomic analysis (Mutoh, Bito, Minami, Nakamura, Honda, Izumi, Nakata, Kurachi, Terano and Shimizu (1993) FEBS Lett. 322, 129-134). The PAF-induced calcium response and phosphoinositide turnover were decreased by pertussis toxin (PTX) treatment, suggesting that these signals are coupled largely with PTX-sensitive G-protein(s) in EoL-1 cells. These systems may provide a useful experimental model with which to investigate the relationship between eosinophilic differentiation and PAF receptor induction, and the role of eosinophils in allergic responses.

[Changes in hemodynamics and mixed venous oxygen saturation during intraoperative harvest of autologous platelet-rich plasma].

Intraoperative harvest and transfusion back of platelet-rich plasma (PRP) are an effective procedure to decrease the requirement of blood transfusion. We selected the right atrium as the site for collection and transfusion back of the blood for the harvest of PRP. We studied changes of the hemodynamics and mixed venous oxygen saturation (SvO2 during the intraoperative harvest of autologous PRP in two patients who were undergoing cardiopulmonary bypass (CPB) surgery. The whole blood was collected, processed and the red blood cells were transfused immediately back to the patients using the Plasma Collection System (Haemonetics Model 50). The procedure was accomplished prior to starting CPB. The PRP product was transfused back into the patients at the completion of CPB. The arterial blood pressure, cardiac output, and SvO2 decreased during the collecting phase and immediately returned to the baseline values during the returning phase. No change was observed in the heart rate and arterial oxygen saturation. We conclude that the usage of right atrium as the site for collection and transfusion back of the blood to harvest PRP is effective and useful, and the continuous monitoring of SvO2 is mandatory for detecting the decline of the cardiac output.

Food consistency modulates eating volume and speed through brain histamine in rat.

Changes in meal parameters of rats fed with different consistency of food were examined using hard and soft pellets. Meal size and eating speed of the first meal after 1800 h increased significantly in rats fed with soft pellets compared to those fed with hard pellets. Effects of histamine depletion on meals treated with hard or soft pellets were investigated after an intraperitoneal injection of 0.11 mmol/kg alpha-fluoromethylhistidine (FMH), a specific suicide inhibitor of the histamine synthesizing decarboxylase enzyme. When rats were fed with hard pellets, FMH significantly decreased eating speed and prolonged meal duration without affecting meal size. When rats were fed with soft pellets, FMH increased meal size and duration, but not eating speed. The meal parameter of eating speed was significantly decreased and meal size and duration were increased in obese Zuckers, a hereditary histamine-depleted animal model, when compared to their lean littermates. These results indicate that proprioceptive sensation from the oral cavity may regulate meal parameters through histaminergic neurons in the brain.

Transfer of innovation for advancement in dentistry: a case study on pit and fissure sealants' use in Japan.

The implementation of scientific innovations is most important in advancing both dental care systems and dental science. However, scientific knowledge in dentistry is not always well implemented. In this paper the phases of development, diffusion, adoption and utilization of sealants are examined as one example of the way in which a relatively new innovation still needs more effort to ensure its wider use. Discussions with a dental manufacturer on the development of sealant materials and an investigation into the adoption of sealants in the national health insurance system of Japan were undertaken. Questionnaires regarding the education on sealants offered in dental schools and dental hygienist schools, and questionnaires to dental practitioners concerning their use of sealants, were used and analysed. The analysis of the various factors showed that the transfer of knowledge through the educational system is the most significant step in the uptake of innovations. Legality, insurance coverage and patient acceptance are significantly associated with the phase of adoption. A model of transfer pathways for new dental innovations is proposed.

[Clinical study on the effect of adjuvant immunochemotherapy using OK-432 and 5-FU in pharyngeal and laryngeal cancers].

A randomized controlled trial was carried out on 38 pharyngeal and 59 laryngeal previously untreated cancers to evaluate the effects of adjuvant immunochemotherapy using 5-FU with and without OK-432. After each fundamental therapy, 5-FU was given orally at a dose of 200-300 mg/day for more than a year. OK-432 was intracutaneously administered at a dose of 5KE once a week for more than a year. Three-year survival rates and disease-free intervals were estimated for the two groups. In cases of pharyngeal cancer, the 3-year survival rate was 58% in both groups and the disease-free interval rates for up to 3 years after the fundamental therapy was 49% in the OK-432 + 5-FU group and 52% in the 5-FU group. These results showed no statistically significant difference. In laryngeal cancers, the 3-year survival rate was 96% in the OK-432 + 5-FU group and 82% in the 5-FU group. These results also showed no statistically significant difference. The disease-free interval rates were 87% in the OK-432 + 5-FU group and 64% in the 5-FU group, revealing a statistically significant difference (p less than 0.1). This suggests that OK-432 is more effective in producing a higher survival rate in cases of laryngeal cancer.