?-arbutin is biosynthesized by whole cell method with Xanthomona maltophilia BT-112.The conditions for cell biosynthesized ?-arbutin are investigated as follows:temperature,25℃;concentration of hydroquinone,30mmol/L;mol ratio of sucrose and hydroquinone,20∶1;time course of ?-arbutin biosynthesis,45 hours;rotational speed,160r/min;concentration of Xanthomona maltophilia BT-112,85g/L;concentration of K-2HPO-4-KH-2PO-4 buffer solution,25mmol/L;pH of K-2HPO-4-KH-2PO-4 buffer solution,8.0.Under the above optimal conditions,the maximum of molar conversion yield based on the amount of hydroquinone supplied reaches 86.7%.

0.05).The setting time of hydroxyaptite and glass ions cements with Co-F were longer but there was little effect on zinc phosphate cements. Conclusion The Co-F agent added to dental cement can not only improve the compressive strength but also contin- ually release fluoride.

Objective To detect the characteristics of benign infantile seizures.Methods Fifty seven cases of benign infantile seizures were analyzed.Results All patients had a normal development before and after the onset of the seizures.The age of onset was from 1.5-30 months.The main manifestations included tonic clonic seizure,staring and motion arrest,64.9 % occurred in clusters.A family history of epilepsy or febrile seizures was present in 6 cases.Interictel electroen cephalograms were normal.The seizures were easily ceased after taking phenobarbital,carbamazepine or valproate.Antiepileptic drugs(AEDs)were discontinued in 51 patients.The mean ages of seizure stopping were 12.8 months and mean seizure′s durations were 4.1 months.Conclusion The benign infantile seizures can be easily controlled with a single AEDs for short time with favorable prognosis.

Aim To explore the lipid-lowering mecha-nisms of curcumin from the molecular levels and pro-vide scientific basis for clinical development of lipid-lowering drugs.Methods Using oil red O staining and enzymic to determinate the levels of cholesterol in HepG2 cells.Moreover,uptaking of DiI-LDL was also measured.The expressions of mRNA and protein were detected by RT-Q-PCR and Western blot.Results The red lipid droplets and the levels of TC and FC sig-nificantly increased in HepG2 cells after treated with curcumin.The orange red fluorescence was higher than that of control.Curcumin could promote the expression levels of mRNA and protein of SREBP2 and LDLR, what′s more,curcumin could reduce the expression of the mature PCSK9 level and IDOL protein.Conclu-sion Curcumin accelerates LDL-C uptake probably via downregulating the expression of PCSK9 and IDOL in HepG2 cells.

The roots of Glycyrrhiza uralensis are widely used in Chinese medicine for their action of clearing heat, detoxicating, relieving cough, dispelling sputum and tonifying spleen and stomach. The reason why Glycyrrhiza uralensis has potent and significant actions is that it contains various active secondary metabolites, especially glycyrrhizic acid. In the present study, we cloned the cDNA coding 3-hydroxy-3-methylglutary CoA reductase (HMGR) involved in glycyrrhizic acid biosynthesis in Glycyrrhiza uralensis. The corresponding cDNA was expressed in Escherichia coli as fusion proteins. Recombinant HMGR exhibited catalysis activity in reduction of HMG-CoA to mevalonic acid (MVA) just as HMGR isolated from other species. Because HMGR gene is very important in the biosynthesis of glycyrrhizic acid in Glycyrrhiza uralensis, this work is significant for further studies concerned with strengthening the efficacy of Glycyrrhiza uralensis by means of increasing glycyrrhizic acid content and exploring the biosynthesis of glycyrrhizic acid in vitro.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Glycyrrhiza uralensis ; enzymology ; genetics ; Glycyrrhizic Acid ; metabolism ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Mevalonic Acid ; metabolism ; Phylogeny ; Plant Roots ; enzymology ; Plants, Medicinal ; enzymology ; genetics ; Recombinant Proteins ; genetics ; metabolism

OBJECTIVETo reveal the 3-hydroxy-3-methylglutary-coenzyme A reductase (HMGR) gene polymorphism of Glycyrrhiza uralensis, and the correlation between HMGR gene polymorphism and the content of glycyrrhizic acid.

METHODLiquorice plants containing different content of glycyrrhizic acid were used as materials. RT-PCR was used to amplify their HMGR gene sequences, which were connected with vector pMD19-T for clone sequencing. Multiple alignments were performed to analyse HMGR gene polymorphism of G. uralensis. Then the correlation between HMGR gene polymorphism and the content of glycyrrhizic acid was revealed.

RESULTHMGR gene sequences polymorphism included codon mutation, base substitution mutation, copy number polymorphism and allele heterozygosity. There were 4 types of mutations in HMGR gene coding amino acid sequences, namely -HSL, -HSV, GALLV, GALSV. Among them, -HSV type was common in liquorice plants, -HSL type only existed in liquorice plants with low content of glycyrrhizic acid, and GALSV type only existed in liquorice plants with high content of glycyrrhizic acid.

CONCLUSIONHMGR gene sequences of G. uralensis are highly polymorphic and related to the content of glycyrrhizic acid.

Cloning, Molecular ; DNA, Complementary ; chemistry ; classification ; genetics ; Glycyrrhiza uralensis ; enzymology ; genetics ; metabolism ; Glycyrrhizic Acid ; metabolism ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Isoenzymes ; genetics ; metabolism ; Mutation ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Polymorphism, Genetic ; Sequence Analysis, DNA

OBJECTIVETo analyse the effect of expression proteins containing different escherichia coli of 3-hydroxy-3-methylglutary-coenzyme A reductase(HMGR) genic mutation on the conversion efficiency of MVA with GC-MS method, in order to lay a foundation for revealing the function of HMGR gene polymorphism of Glycyrrhiza uralensis in the production of high-quality G. uralensis medicines.

METHODThe expression carrier was established from four HMGR genic mutation types cloned from G. uralensis and transformed into Escherichia coli BL21. The protein was induced to express, detected and purified. The purified protein was adopted for in vitro enzymatic reaction. TLC and GC-MS were used for qualitative and quantitative analysis on reaction products.

RESULTThe catalytic activity of L/V genotype(-HSL and -HSV) was similar, and so was the catalytic activity of the genotype with GA insertion (GALLV and GALSV), but the catalytic activity of the latter was around 2 times higher than that of the former.

CONCLUSIONThe functional gene polymorphism of G. uralensis may be the molecular foundation for the production of high-quality G. uralensi medicines.

Biocatalysis ; Chromatography, Thin Layer ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Gas Chromatography-Mass Spectrometry ; Glycyrrhiza uralensis ; enzymology ; genetics ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Mutation ; Plant Proteins ; genetics ; metabolism ; Polymorphism, Genetic ; Recombinant Proteins ; metabolism

OBJECTIVETo explore the metabolism-related risk factors of cholelithiasis among residents in Beijing.

METHODSThe clinical data including previous disease history, findings of physical examination, and results of cholecystosonography of 2270 patients with cholelithiasis identified in the Health Screening Center of Peking Union Medical College Hospital between August 2007 and August 2010 were retrospectively reviewed (the case group). Meanwhile, 4336 healthy individuals during the same period were randomly chosen as the control group.

RESULTSTotal cholesterol, triglyceride, low-density lipoprotein cholesterol, fasting blood glucose, body mass index, and systolic blood pressure were positively correlated with the incidence of cholelithiasis (P < 0.05), while high-density lipoprotein cholesterol was negatively correlated (P < 0.05). Diastolic blood pressure showed no association with cholelithiasis (P > 0.05).

CONCLUSIONCholelithiasis is resulted from multiple factors including elevated blood lipids, blood glucose, and systolic blood pressure among residents in Beijing.

Adult ; Aged ; Blood Glucose ; Blood Pressure ; Case-Control Studies ; China ; epidemiology ; Cholelithiasis ; epidemiology ; metabolism ; Female ; Humans ; Lipids ; blood ; Male ; Middle Aged ; Risk Factors

OBJECTIVETo observe the effect of Coptidis Rhizoma (CR) on hemolysis and antioxidant system of normal mice and its impact on the functions, while evaluating the oxidation reduction property of CR and berberine.

METHODIn the whole animal experiment, normal mice were orally administered with CR at the dose of 1.2 g x kg(-1) for three days. Their blood were collected to detect the hemoglobin in plasma, the content of serum bilirubin, the number of peripheral blood reticulocytes, the T-AOC in whole blood, measure the contents of glucose-6-phosphate-dehydrogenase (G6PD), superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) of RBC membrane, determine the activity of Na(+)-K(+)-ATPase, Ca(2+)-Mg(2+)-ATPase, fluidity, and observe its impact on the liquidity and deformability of RBCs. According to the electrical and biochemical experiment, the voltammetric behaviors of CR and berberine on glassy carbon electrode were evaluated using cyclic voltammetry. In the RBC in vitro experiment, the impact of Coptidis Rhizoma on autoxidation hemolysis rate of RBCs of normal mice was observed.

RESULTThere was no significant effect on hemoglobin, serum bilirubin, and reticulocyte count in normal mice administrated with CR at the dose of 1.2 g x kg(-1), and so is on RBC membrane SOD, G6PD, MDA, GSH and whole blood T-AOC activity. In addition, CR had also no significant effect on Na(+)-K(+)-ATPase, Ca(2+)-Mg(2+)-ATPase activity, and no notable impact on the fluidity and deformability of RBCs. There were two oxidation peaks at -0.27 V and 0.60 V induced by CR and one oxidation peak induced by berberine at 0.56 V, with no reduction peak at fly-back. CR could significantly inhibit oxidative hemolysis in RBCs at the dose of 0.125-2 g x L(-1) in vitro.

CONCLUSIONThe normal dose of Coptidis Rhizoma can not cause hemolysis of RBC, and also can not change antioxidant system and functions of RBC, CR and berberine show antioxidant (reducing) properties.

Animals ; Antioxidants ; pharmacology ; Berberine ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Erythrocytes ; drug effects ; metabolism ; Female ; Hemolysis ; drug effects ; Male ; Mice

OBJECTIVETo examine changes of blood oxidative-antiovidative level in schizophrenic patients and its relationship with clinical symptoms.

METHODSForty-six Chinese patients met DSM-IV (Diagnostic and Statistical Manual of Mental Disorders-IV) criteria for schizophrenia and fifty age- and sex-matched healthy controls were enrolled in the present study. Baseline psychiatric symptom severity was assessed with brief psychiatric rating scale, positive and negative syndrome scale on the blood draw day. Fresh blood samples were collected to measure levels of nitric oxide and lipid peroxide in plasma as well as activities of superoxide dismutase, catalase and glutathione peroxidase in red blood cells by spectrophotometric assays simultaneously.

RESULTSComparison of the biochemical parameters indicated that the level of nitric oxide and lipid peroxide increased in patient group, which represented a positive correlation with positive scale scores; while the activities of three critical enzymes decreased and showed a negative linear correlation.

CONCLUSIONThis study showed that there are dysregulation of free radical metabolism and poor activities of the antioxidant defense systems in schizophrenic patients. Excess free radicals formation may play a critical role in the etiology of schizophrenia. Using antioxidants might be an effective therapeutic approach to partially alleviate or prevent the symptoms of schizophrenia.

Adolescent ; Adult ; Antioxidants ; metabolism ; Female ; Free Radicals ; Humans ; Lipid Peroxides ; metabolism ; Male ; Middle Aged ; Nitric Oxide ; metabolism ; Schizophrenia ; etiology ; metabolism