Several kinds of column chromatography method were used to investigate the chemical constituents of the leaves of Lophatherum gracile. The structures of the isolated compounds were identified based on their physicochemical properties and spectral data. A new flavone C-glycoside was isolated and its structure was identified as 3'-methoxyl-luteolin 6-C-beta-D-galactopyranosiduronic acid (1 --> 2) -alpha-L-arabinopyranoside (1).
Antiviral Agents ; chemistry ; isolation & purification ; Flavones ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Hydrolysis ; Plant Leaves ; chemistry ; Poaceae ; chemistry

To investigate the effect of flavonoids from Sophora flavescens in aging mice induced by D-galactose and its mechanism. Totally 60 mice were randomly divided into six groups: the control group, the model group, the piracetam group (positive control group) and flavonoids from S. flavescens low, medium and high doses groups. Except for the control group, all of the rest groups were subcutaneously injected with D-galactose (160 mg x kg(-1)) for successively 30 days to establish the sub-acute senescent model. Meanwhile, flavonoids from S. flavescens low, medium and high doses groups were respectively administered with 150, 300 and 600 mg xkg-('1)of flavonoids from S. flavescens for 30 days. The learning and memory abilities of mice were determined by avoiding darkness ex-eriment and jumping stair experiment. The contents of malondialdehyde (MDA) tumor necrosis factor-aα NF-aα the activities of superoxide dismutase (SOD) monoamine oxidase-B (MAO-B) Na'(+)K'(+)-ATPase and Ca2(+ )-ATPase in the brain of mice were deter-ined respectively after the behavioral experiments. The activity of lactic dehydrogenase ( DH) in blood serum was also determined and analyzed by microscope after HE staining to observe the changes in hippocampal organizational structure. Compared with the model group, flavonoids from S. favescens medium and high doses groups showed significantly increases in the latency of avoiding darkness and jumping stair experiments; flavonoids from S. fllvescens low, medium and high doses groups and the piracetam group showed de-reases in the numbers of errors in avoiding darkness experiment; the flavonoids from S. flavescens high dose group and the piracetam group showed reduction- n the number of errors in jumping stair experiment (P <0 . 5 or P <0 . 1). Flavonoids from S. flavescens me-ium and high doses groups and the piracetam group showed improvements in the activities of SOD, Na'(+)K'(+)ATPase in the brain of mice and declines in the contents of MDA and TNF-aα the activity of MAO-B in the brain of mice, the activity of LDH in blood serum (P <0 . 5 or P <0 . 1). Flavonoids from S. flavescens low, medium and high doses groups and the piracetam group also showed im-rovement in the activity of Ca2(+ )ATPase, with statistical difference from the model group (P <0 . 5 or P <0 . 1). The pathological result showed decreases in the number of cells of hippocampal dentate gyrus area, sparse cell arrangement, incomplete cellular mor-hology, scarce cytoplasm, blurred boundary between nucleus and cytoplasm, nuclei anachromasis, irregular pyknosis and unconspicu-us nucleoli in the model group. Compared with the model group, flavonoids from S. flavescens low, medium and high doses groups and the piracetam group showed improvements in hippocampal organization tissues. Flavonoids from S. favescens can improve the learning and memory ability of senescent mice induced by D-galactose. Its mechanism may be correlated with the enhancement of anti-oxidative actions by lowering TNF-aαcontent, which results in the stability of cell membrane and the reduction in MAO-B activity.
Aging ; drug effects ; metabolism ; psychology ; Animals ; Brain ; drug effects ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Flavonoids ; administration & dosage ; Galactose ; adverse effects ; Hippocampus ; drug effects ; metabolism ; Humans ; Learning ; drug effects ; Male ; Malondialdehyde ; metabolism ; Mice ; Sophora ; chemistry ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Little is known about the non-structural protein 6(NSP6)of rotavirus.This report describes expression of the NSP6 of a group A human rotavirus strain TB-Chen in bacteria,and its immunological properties and cellular distribution.The results showed that the recombinant NSP6(rNSP6)was expressed in high efficiency without any other proteins fused(possesses about 34.2% of total bacterial proteins).rNSP6 elicited mono-specific antibodies in immunized guinea pigs and the antibodies could react with the rNSP6 itself and the viral NSP6 proteins synthesized in SA11-or Wa-infected MA104 cells in Western blot and immunofluorescence assay.The NSP6 distributed evenly in the cytoplasm mainly around the nucleus of virus-infected cells,no viroplasm-like gathering observed;The top amount of NSP6 synthesized in SA11-infected cells or Wa-infected cells could be detected at 12h after infection.This is the first report about the high expression of entire NSP6(without any other proteins fused)in prokaryotic expression system and detection of NSP6 synthesis in virus infected cells by immunofluorescence assay.The results are important to understand the structure,biological properties and further application of the NSP6.

Twenty Newcastle disease virus(NDV)strains were isolated from diseased chicken and geese in field outbreaks during 2005 and 2006 in some regions of Jiangsu and Guangxi,and the antigenic analysis of the all NDV isolates had been done based on the reaction spectrum with a panel of monoclonal antibodies to the HN glycoprotein.The entire ORFs encoding HN protein of these NDV isolates were amplified by RT-PCR successfully,cloned and sequenced.The resultant sequences of HN genes of 13 isolates of chicken origin and 7 isolates of goose origin were gained and analyzed.The results of reaction spectrum showed that there were some distinct differences in the antigenic epitopes among the 20 NDV isolates.And the sequences revealed that the coding regions of the HN genes of these isolates all consisted of 1716 nt characteristic of virulent strains of NDV,coding for 571 amino acids.Neucleotides sequence homology were found to be from 94.8%to 100%among 18 NDV isolates of genotypeⅦ,and the neucleotides sequence homology between all the isolates and the other genotypeⅦstrains of recent years in China ranged from 92.1%to 99.6%.The deduced amino acid sequences and the receptor-binding regions of HN proteins between the NDV isolates of chicken origin and of goose origin were compared and analyzed.The results showed that some unique amino acid substitutions were found in the genome of the NDV isolates,and the close genetic similarity provided evidence for epidemiological linkage between the NDV isolates of chicken origin and of goose origin in the same period.

OBJECTIVETo investigate the role of human angiopoietin-1 (Ang1) in tumorigenesis and angiogenesis of human gastric cancer cell line SGC7901 in nude mice.

METHODSRecombinant human Ang1 sense or antisense eukaryotic expression vectors were constructed, and transfected by lipofectin into human gastric cancer line SGC7901. Stable transfectants were obtained respectively, namely 7Ang1+ for sense, 7Ang1- for antisense, and 7901P for empty vector transfected cells. Semiquantitative PCR and Western blot were employed to testify the transfection efficiency. Cell growth curve and cell cycle were observed by MTT assays or flow cytometry. In in vivo study, growth of SGC7901 xeno-transplant was observed in BALB/c nude mice. Microvessel density (MVD) was analyzed by immunohistochemistry for Factor VIII staining.

RESULTSStably transfected cell lines were established and decreased expression of Ang1 protein and mRNA in the antisense transfected SGC7901 cells was achieved. Tumorigenesis of 7Ang1- cells on day 30 days was significantly inhibited with decreased MVD as compared to that in 7901P and 7Ang1+ cells (P < 0.01).

CONCLUSIONAngiopoietin-1 plays an important role in tumorigenesis and angiogenesis of gastric cancer which can be partially abrogated by antisense technique.

Angiopoietin-1 ; biosynthesis ; genetics ; Animals ; Cell Line, Tumor ; DNA, Antisense ; genetics ; Genetic Vectors ; Humans ; Mice ; Mice, Nude ; Microcirculation ; pathology ; Neoplasm Transplantation ; Neovascularization, Pathologic ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Neoplasms ; blood supply ; metabolism ; pathology ; Transfection

Increased vascular endothelial growth factor (VEGF) biosynthesis in vascular endothelial cells has been reported to play an obligatory role in promoting angiogenesis. Nevertheless, the intracellular signaling mechanisms of hypoxia-induced VEGF release remain largely unknown. Human umbilical vein endothelial cell lines (ECV304) were cultured in normoxic or hypoxic conditions for 12 approximately 24 h and harvested for determination of VEGF mRNA expression and phosphorylation of ERK1/2 and p38 mitogen-activated protein kinase (p38 MAPK) by real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Secreted VEGF protein was measured by enzyme-linked immunosorbent assay (ELISA). It has reported that PD98059, an ERK inhibitor, was able to blunt the hypoxia-induced activation of the expression of VEGF gene. In accordance with this report, an increase in ERK1/2 phosphorylation and VEGF biosynthesis was observed in ECV304 cells cultured in hypoxia, and this increase was blocked by PD98059. The novel finding of the present study is that an activation of p38 MAPK is involved in hypoxia-induced increase in VEGF biosynthesis. SB202190, an inhibitor of p38 MAPK was able to blunt the hypoxia-induced increase in VEGF biosynthesis. These dada provide the first direct evidence for a role of p38 MAPK in mediating hypoxia-induced increase in VEGF biosynthesis in human endothelial cells.
Cell Hypoxia ; Cell Line ; Endothelial Cells ; cytology ; metabolism ; Humans ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; RNA, Messenger ; genetics ; metabolism ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism ; physiology

The purpose of this study was to explore the effect of amifostine in treating aged patients with idiopathic thrombocytopenic purpura (ITP). Three aged ITP patients (one aged 88, one aged 75, another aged 65) were treated with amifostine over a period of 4 weeks for each. The results showed that the early positive responses were observed in all 3 aged patients after treatment with amifostine 5 x 400 mg per week for 4 weeks, the recovery of platelet counts to normal level were found in 2 cases after 1 week and in 1 case after 2 weeks. Following 4 months after stop of treatment with amifostine, the normal platelet counts still remains in all patients, no more blood transfusions, hormonal preparation and gamma-globulin were given. Amifostine has never been reported elewhere as to treat of aged ITP. In conclusion, amifostine has a good early therapeutic effect on aged ITP patients, but its clinical outcome of long-term still needs further evaluation.
Aged ; Aged, 80 and over ; Amifostine ; therapeutic use ; Drug Administration Schedule ; Humans ; Male ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; Treatment Outcome

The objective of this study was to explore the effect of amifostine (AMF) in treating patients with idiopathic thrombocytopenic purpura (ITP) and its adverse effects. 17 ITP patients (aged from 13 to 92, mean 65 years) were treated with AMF and their therapeutic regimen was following: 0.4 g AMF was dissolved in 100 ml physiological saline, the intravenous drop was performed at day and lasted 5 days per week, with interval 2 days, 4 weeks were considered as a course of treatment. The results showed that the normal counts of platelets were observed in 17 ITP patients after 1 course of treatment, and remained unchanged for 2 months following stop of therapy. In that time, no any drugs such as hormonal preparation, gamma-globulin, blood transfusion and so on were given in all treated patients, the main adverse effect of AMF was discomfort in digestive system that all patients could endure. Therefore, the patient's life quality was obviously improved. It is concluded that the AMF has been initially and successfully used to treat ITP patients with positive response, and may be considered as a safe-effective drug for treating old patients with ITP, but the long-term effect and pharmacological mechanism of this drug needs further evaluation.
Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Amifostine ; adverse effects ; therapeutic use ; Female ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; Treatment Outcome

The essence of endogenous turbidity in Chinese medicine (CM) is different from cream, fat, phlegm, retention, damp, toxicity, and stasis. Along with the development of modern scientific technologies and biology, researches on the essence of endogenous turbidity should keep pace with the time. Its material bases should be defined and new connotation endowed at the microscopic level. The essence of turbidity lies in abnormal functions of zang-fu organs. Sugar, fat, protein, and other nutrient substances cannot be properly decomposed, but into semi-finished products or intermediate metabolites. They are inactive and cannot participate in normal material syntheses and decomposition. They cannot be transformed to energy metabolism, but also cannot be synthesized as executive functioning of active proteins. If they cannot be degraded by autophagy-lysosome or ubiquitin-prosome into glucose, fatty acids, amino acids, and other basic nutrients to be used again, they will accumulate inside the human body and become endogenous turbidity. Therefore, endogenous turbidity is different from final metabolites such as urea, carbon dioxide, etc., which can transform vital qi. How to improve the function of zang-fu organs, enhance its degradation by autophagy-lysosome or ubiquitin-prosome is of great significance in normal operating of zang-fu organs and preventing the emergence and progress of related diseases.
Autophagy ; Humans ; Medicine, Chinese Traditional ; Proteasome Endopeptidase Complex

Adenocarcinoma ; blood supply ; physiopathology ; Capillaries ; metabolism ; ultrastructure ; Cell Separation ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; ultrastructure ; Humans ; Microcirculation ; Stomach Neoplasms ; blood supply ; physiopathology