The diethyl sulfate (DES) mutagenesis was chosen for the mutagenic treatment to Phellinus igniarius, and the relationship of mutagenesis time and death rate was investigated with 0.5% DES. The differences of mycelial growth speed, liquid fermentation mycelia biomass, morphology and pigment classes of secondary metabolites production speed and antioxidant activities of metabolite products were discussed. The study displayed that DES mutagenesis could change mycelial morphology without obvious effect on mycelium growth, and the DES mutagenesis improved antioxidant activities of the active ingredients of P. igniarius and had more antioxidant activity of hypoxia/sugar PC12 nerve cells than that of P. igniarius.
Basidiomycota ; drug effects ; genetics ; growth & development ; metabolism ; Mutagenesis ; Mutagens ; pharmacology ; Mycelium ; drug effects ; genetics ; growth & development ; metabolism ; Pigments, Biological ; analysis ; metabolism ; Secondary Metabolism ; drug effects ; Sulfuric Acid Esters ; pharmacology

OBJECTIVETo obtain the full-length cDNA of a novel gene (named yp05) associated with citrinin production-related genes in Monascus aurantiacus.

METHODSTotal RNA was extracted from mycelium, 3' and 5' cDNA end of yp05 gene was amplified using smart trace cDNA amplification kit, and the full-length cDNA of a novel gene (named yp05) was obtained from the electronic assembly of 3'-RACE and 5'-RACE products.

RESULTSThis yp05 gene was 787 bp including a 597 bp open reading frame (ORF) and encoded a deduced protein with 199 amino acid residues, and the amino acid sequence of this protein was found similar with the sequences of many fungal manganese-superoxide dismutases in the GenBank with the aid of BLASTp. The transcription of yp05 gene in Monascus strains was analyzed with the aid of Northern blotting. The transcription of yp05 gene was only detected in Monascus strains, provided that citrinin was produced.

CONCLUSIONThe transcription of yp05 gene belongs to differential expression genes of citrinin yielded from Monascus and has no correlation with the biosynthesis pathway of red pigments.

Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; Citrinin ; biosynthesis ; Cloning, Molecular ; DNA, Complementary ; chemistry ; Fungal Proteins ; chemistry ; genetics ; Gene Library ; Molecular Sequence Data ; Monascus ; genetics ; metabolism ; Mycelium ; genetics ; metabolism ; Pigments, Biological ; biosynthesis ; RNA, Messenger ; metabolism ; Sequence Alignment ; Sequence Analysis, DNA

OBJECTIVEThis study is to investigate the effect of tea polyphenols and tea pigments on telomerase activity of human liver cancer cell line, HepG2 cells.

METHODSTRAP-PCR-ELISA was applied to investigate the telomerase activity.

RESULTSTelomerase was positive in tea polyphenols treated groups, tea pigments treated groups and blank control group. Telomerase activities (A(450 approximately 690) values) were 1.56 and 1.46 in 50 mg/L and 100 mg/L tea polyphenols-treated groups, 1.55 and 1.49 in 50 mg/L and 100 mg/L tea pigments-treated groups, respectively. The results showed that telomerase activity was significantly inhibited by tea polyphenols and tea pigments treatment as compared with the blank control group (A(450 approximately 690) = 2.11).

CONCLUSIONSTea polyphenols and tea pigments could significantly inhibit telomerase activity of HepG2 cells, and telomerase activity may be a useful biomarker for cancer chemoprevention.

Biomarkers, Tumor ; metabolism ; Carcinoma, Hepatocellular ; enzymology ; pathology ; Flavonoids ; isolation & purification ; pharmacology ; Humans ; Liver Neoplasms ; enzymology ; pathology ; Phenols ; isolation & purification ; pharmacology ; Pigments, Biological ; isolation & purification ; pharmacology ; Polyphenols ; Tea ; chemistry ; Telomerase ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo find out the differences between 102 kb pgm locus of Yersinia pestis isolated from Microtus brandti with of other types, and the characters of Yersinia pestis isolated from Microtus brandti caused by their makeup of the 102 kb pgm locus.

METHODS102 kb pgm locus of Yersinia pestis isolated from Microtus brandti and Yersinia pestis isolated from Marmota himalayana were amplified by polymerase chain reation (PCR) with 25 pair of nested primers. The PCR products of one pair of primer were obviously different, and then cloned and sequenced. Sequences were searched against current protein and nucleotide databases, using BLAST.

RESULTSThe 102 kb pgm locus of Yersinia pestis isolated from Microtus brandti was devoid one IS100. In addition, it had more copies than other types in the similar variable-number tandem repeat sequences.

CONCLUSIONThe 102 kb pgm locus of Yersinia pestis was different from that of other types. It had only one IS100 flanked it, which corresponded to the character that its pgm(+) phenotype was stable. Further study was needed to confirm the relationship between the diminution virulence of Yersinia pestis isolated from Microtus brandti and the loss of IS100 and other changes.

Animals ; Arvicolinae ; microbiology ; Bacterial Proteins ; genetics ; metabolism ; China ; epidemiology ; Cloning, Molecular ; DNA, Bacterial ; Humans ; Pigments, Biological ; genetics ; Plague ; epidemiology ; microbiology ; Polymerase Chain Reaction ; Repetitive Sequences, Nucleic Acid ; Sequence Analysis, DNA ; Virulence ; genetics ; Yersinia pestis ; genetics ; pathogenicity

OBJECTIVETo investigate the inhibitory effect of lycium pigment on lipopolysaccharide (LPS)-induced uveitis in rats and its mechanism.

METHODThe rat uveitis model was established by 30-day oral administration of lycium pigment (50, 100, 200 mg x kg(-1)) and footpad injection of LPS. Ocular tissues were collected for a histopathological inspection. The protein, nitric oxide and ADMA in aqueous humor, level of inducible nitric oxide synthase (iNOS) in retina, activities of serum total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX), and content of malondialdehyde (MDA) were determined by using Western blot, ELISA and biochemical methods.

RESULTAccording to the pathological study, lycium pigment (50, 100, 200 mg x kg(-1)) could notably reduce the inflammatory cell infiltration around corpus ciliare matrix of uveitis rats, and the concentration of protein and nitric oxide, and increased ADMA in aqueous humor. Lycium pigment (100, 200 mg x kg(-1)) could significantly inhibit the expression of iNOS in ocular tissues. In addition, lycium pigment (100, 200 mg x kg(-1)) also decrease the activities of serum T-AOC, SOD, GSH-PX, and the content of lipid peroxide MDA.

CONCLUSIONLycium pigment has the inhibitory effect on LPS-induced uveitis in rats. Its mechanism is related to the regulation of nitric oxide/ADMA pathway and the improvement of oxidation resistance.

Animals ; Glutathione Peroxidase ; genetics ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Lycium ; chemistry ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Pigments, Biological ; administration & dosage ; Plant Extracts ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; genetics ; metabolism ; Uveitis ; chemically induced ; genetics ; metabolism ; prevention & control

S-adenosylmethionine-dependent uroporphyrinogen III methyltransferase (SUMT) is a novel red fluorescence indicator. However, the production of SUMT in Escherichia coli is restricted by its relatively low solubility, and little is known about the red fluorescent materials that are associate with SUMT. Two individual SUMT mutations, L166A and L88R/L89G double mutant were produced by site-directed mutagenesis. Both mutants were overexpressed in E. coli and purified by Ni-NTA chromatography. The reddish mixtures isolated from the purified L88R/L89G double mutant were analyzed by UV-visible spectra scanning and mass analysis(MS). The L88R/L89G double mutant has enzymatic activity in vivo, whereas L166A mutant loses the activity. Trimethylpyrrocorphin is identified as the main constituent in the isolated pigments. The purified L88R/L89G mutant increases protein solubility, which is applied potentially as the fluorescent indicator denoting the solubility of protein fusion partner.
Amino Acid Substitution ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Fluorescence ; Mass Spectrometry ; Methyltransferases ; chemistry ; genetics ; metabolism ; Molecular Weight ; Mutagenesis, Site-Directed ; methods ; Mutation ; Pigments, Biological ; chemistry ; metabolism ; Plant Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; chemistry ; metabolism ; Solubility ; Spectrophotometry, Ultraviolet ; Zea mays ; enzymology ; genetics

OBJECTIVETo elvaulate the antioxidant activity of the pigment of Lycium ruthenicum.

METHODThe antioxidant activities were measured by the effects of the reducing ability, scavenging DPPH. H2O2-induced hemolysis of mice erythrocyte, serum resistance of reactive oxygen species, content of MDA in liver tissue, and swelling effect of mitochondria in liver tissue.

RESULTThe pigment of L. ruthenicum could scaveng DPPH* remarkably with IC50 0.164 mg x mL(-1), inhibitte hemolysis of mice erythrocyte evidently with IC50 0.112 mg x mL(-1). The resistant of reactive oxygen species was enhanced by the tested substances, simultanously. The concentration of MDA of peroxidation of lipid in mice liver could be reduced, and the swelling of mice liver mitochondria alse be restrained.

CONCLUSIONThe pigment of L. ruthenicum has antioxidant activity in tested concentration.

Animals ; Antioxidants ; pharmacology ; Biphenyl Compounds ; metabolism ; Erythrocytes ; drug effects ; Free Radical Scavengers ; pharmacology ; Hemolysis ; drug effects ; Hydrazines ; metabolism ; Lipid Peroxidation ; drug effects ; Liver ; metabolism ; Lycium ; chemistry ; Malondialdehyde ; metabolism ; Mice ; Mitochondria, Liver ; pathology ; Mitochondrial Swelling ; drug effects ; Phenols ; isolation & purification ; pharmacology ; Picrates ; Pigments, Biological ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Reactive Oxygen Species

No abstract available.
Aged ; Anti-Bacterial Agents/therapeutic use ; Bacteria/metabolism ; Catheter-Related Infections/diagnosis/drug therapy/*microbiology ; Color ; Equipment Design ; Escherichia coli Infections/diagnosis/drug therapy/*microbiology ; Female ; Humans ; Intestines/*microbiology ; Pigments, Biological/metabolism ; Treatment Outcome ; Tryptophan/metabolism ; Urinary Bladder Neoplasms/surgery ; Urinary Catheterization/adverse effects/*instrumentation ; *Urinary Catheters ; *Urinary Diversion ; Urinary Tract Infections/diagnosis/drug therapy/*microbiology ; Urine/chemistry/microbiology

OBJECTIVESThis study is to investigate the effects of tea polyphenols and tea pigments on cell cycle regulators in rat liver precancerous lesions.

METHODSThe modified Solt-Farber precancerous liver rat model was used. Rats were given water, tea polypheol solution (0.1%) or tea pigment solution (0.1%) throughout the whole experiment (56 days). Cyclin D1, P21(WAF1/CIP1), GADD45 and PCNA protein expression were detected by Western blotting and the RT-PCR method was applied to study the expression of Cdk4.

RESULTSCyclin D1, Cdk4 and PCNA expressions were significantly inhibited, and the expression of P21(WAF1/CIP1) and GADD45 were significantly induced by tea polyphenols and tea pigments treatments.

CONCLUSIONTea polyphenols and tea pigments induced cell cycle arrest and inhibited cell proliferation by regulating cell cycle regulators.

Animals ; Blotting, Western ; Cell Cycle Proteins ; drug effects ; genetics ; metabolism ; Cyclin D1 ; drug effects ; metabolism ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p21 ; Cyclin-Dependent Kinases ; genetics ; Cyclins ; drug effects ; metabolism ; Flavonoids ; Intracellular Signaling Peptides and Proteins ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Male ; Phenols ; pharmacology ; Pigments, Biological ; pharmacology ; Polymers ; pharmacology ; Polyphenols ; Precancerous Conditions ; genetics ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; drug effects ; metabolism ; Proteins ; Proto-Oncogene Proteins ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Tea ; chemistry