By functional plates,16 strains which can produce?-mannana-se were isolated frnm 28 Bacillus spp.Using a pair of degenerated primers,the conserved fragments of?-mannanase gene from the selected strains were amplified by PCR.The obtained nucleotide fragments were sequenced and compared with the homologous?-mannanase genes in GenBank and a phylogenetic tree was generated.Comparing to the genes coding?-mannanase published,the cloned nucleotide fragments show the highest sequence identity between 62% and 98%.The genes coding fnr?-mannanase of Bacillus circulus have low identity while the?-mannanase genes of Bacillus subtilis and other Bacillus spp. have high identity.

The Qinbai Qingfei concentrated pellets by traditional Chinese medicine theoryand party and group, the rats were given the drugs group, comparison of pharmacokinetics parameters changes of baicalin , discusses the rationality of Qinbai prescription. The rats were gavaged monarch drug group (Huang Qincu extract, mainly forbaicalin), and official medicine group, adjuvant group, medicine group and Qinbai group (Quan Fangzu) the content of baicalin equal as the monarch drug group, in the 28 h collection in rat plasma at different time point, application of HPLC determination of baicalin glycosides in rat plasmaconcentration time curve, with 3P97 practical pharmacokinetics program to process the data Based on the data analysis, baicalin in rat plasma of Qinbai group Cmax is 4 times as big as monarch druggroup, AUC is 6 times as big as monarch drug group; the content of baicalin in plasma of rats the highest is Qinbai group, the minister drug group, adjuvant group, medicine group of baicalin in rat plasma content of less than the Qinbai group, but was significantly higher than that of monarch drug group; the medicine group is slightly higher than that adjuvant the content of baicalin in plasma of rats. The pharmacokinetic results show that the measured plasma concentration in rats that Qinbai can significantly increase Cmax and AUC of baicalin, other components of qinbai can promoted the baicalin absorption in vivo. It showed that the reasonable of Qinbai compound compatibility. The minister drug can promote the absorption of baicalin in vivo.
Animals ; Area Under Curve ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Flavonoids ; blood ; pharmacokinetics ; Intestinal Absorption ; Male ; Rats ; Rats, Wistar

OBJECTIVETo isolate and identify the pathogen of Dengue fever from Shenzhen city in 2005 - 2006, and to analyze the molecular characteristics of the isolated Dengue virus strain as well as to explore its possible origin.

METHODSIgM and IgG of serum samples taken from 60 suspected Dengue fever patients were detected by ELISA and immunochromatography, and 9 specimens were positive. Nine samples from patients with early stage Dengue fever were used to isolate virus with C6/36 cell line and the positive cell cultures were identified by MGB fluorescent PCR. The type of isolated virus strain was determined by RT-semi-nested-PCR and fluorescent PCR. E gene of isolated virus strain was amplified by RT-PCR and sequenced. Homology and phylogenetic tree of E gene of Shenzhen Dengue virus with the strains isolated from other areas were constructed.

RESULTSOf nine antibody-positive serum samples, one strain of Dengue virus was successfully isolated. The isolated virus strain was confirmed as Dengue virus type 2 and designated as DEN2-SZ0521. The homology of nucleotide sequence and the deduced amino acid sequence of E gene of SZ0521 with standard type 2 Dengue virus NGC strain was 94.2% and 98.2%, but the homology with standard Dengue virus 1, 3, 4 in the same fragment were 59.1%, 57.2%, 58.5% and 68.1%, 66.7%, 63.2%, respectively. The phylogenetic tree indicated that SZ0521 had the greatest similarity with the Malay0412a/Tw strain and they lied in the same branch of the phylogenetic tree. The corresponding homology of nucleotide sequence and amino acid sequence was 99.8% and 100%, respectively. The isolated Dengue virus type 2 belonged to genotype IV with Indonesia-76, Somalia-84 and Sri Lanka-90.

CONCLUSIONDengue virus was isolated from Shenzhen for the first time, and it was classified as type 2. It was confirmed that the type 2 Dengue virus may come from the epidemic area in Malaysia.

Aedes ; virology ; Animals ; China ; Dengue ; virology ; Dengue Virus ; classification ; genetics ; isolation & purification ; Genes, Viral ; Humans ; Phylogeny ; Sequence Analysis, Protein ; Sequence Analysis, RNA

Vestibular rehabilitation has been applied in children with benign paroxysmal positional vertigo, cerebral palsy and central coordination disorder successfully. However, training intensity and methods are various with diseases and therapeutic purposes. This paper reviewed the recovery mechanisms, the assessment and rehabilitation of vestibular dysfunction, and discussed the problems and prospects of vestibular rehabilitation for children.

Objective To evaluate the efficacy and safety of the loratadine paracetamol and pseudoephedrine(LPP)sustained-release tablets in the treatment of common cold and influenza compared with compound pseudoephedrine hydrochloride tablets.Methods Multi-center,randomized,double-blinded controlled clinical trial.One hundred eighteen patients were given LPP sustained-release tablets as trial group,119 were given pseudoephedrine hydrochloride as control group.Results Non-inferiority statistics showed that the improvement rate of clinical symptoms in trial group were not lower than that in control group(P<0.05).The total symptom score did not differ in trial group and control group(P>0.05).The effective rate in trial group and control group were 96.33% and 96.36%(P>0.05).The incidence of adverse drug reactions in trial group and control group was 7.02% and 11.82%,respectively(P>0.05).Conclusion LPP sustained-release tablets is effective and safe in the treatment of common cold and influenza.

The aim of this study was to observe the protective effect of ademetionine 1, 4-butanedisulfonate on liver injury caused by chemotherapy in patients with leukemia. The clinical data of protective effect were analyzed retrospectively from January 2010 to April 2012. A total of 62 acute leukemia patients were divided into A group (27 cases) and B group (35 cases), the polyene phosphatidyl choline combined with ademetionine or combined with compound glycyrrhizin were given in A and B group, respectively. The changes of liver function were observed after 2 weeks, 5 patients in B group suffered from acute liver injury were treated by ademetionine as rescue therapy. Liver function was compared before and after treatment. The results showed that ALT and AST levels were significantly reduced in A group (P < 0.05), none of the patients (0/27) suffered from acute liver injury, but 14.29% (5/35) patients in B group suffered from acute liver injury, and liver function could be recovered by substitution treatment of ademetionine (the median time is 8 days, 5-14 days). It is concluded that the protective and therapeutic effect of ademetionine against liver injury caused by chemotherapy in patients with leukemia is better than that of compound glycyrrhizin.
Adult ; Antineoplastic Agents ; adverse effects ; Chemical and Drug Induced Liver Injury ; prevention & control ; Female ; Glycyrrhizic Acid ; therapeutic use ; Humans ; Leukemia ; drug therapy ; Male ; Middle Aged ; S-Adenosylmethionine ; analogs & derivatives ; therapeutic use ; Young Adult

Animals ; Ginsenosides ; pharmacology ; Hypertension, Pulmonary ; etiology ; metabolism ; Hypoxia ; complications ; metabolism ; Lung ; drug effects ; metabolism ; MAP Kinase Signaling System ; Male ; Panax ; Rats ; Rats, Sprague-Dawley

Aspergillus fumigatus wild-type phytase has many favorable properties, such as a good thermorstability and a broad pH optimum. However, the specific activity of the enzyme is relative low. A. fumigatus Q23L phytase resulted in a remarkable increase in specific activity around pH4.5 - 7.0, but the pH stability of Q23L was lower than A. fumigatus wild-type phytase. To increase the pH stability of Q23L, the mutant Q23LG272E was constructed by site-directed mutagenesis with PCR. The gene of A. fumigatus wild-type phytase and the mutant genes encoding the Q23LG272E and the Q23L were correctly expressed in Pichia pastoris GS115. Enzymes were purified and their enzymatic properties were determined. The results revealed that the specific activity of the Q23L improved remarkably, which increased from 51 u/mg of the wild type to 109 u/mg at pH5.5. Meanwhile, the pH stability of Q23L, decreased evidently, especially from pH3.0 to pH4.0.The pH stability of Q23LG272E in pH3.0 - 4.5 and pH6.5 - 7.0 has been improved compared with Q23L. The specific activity of Q23LG272E basically maintained at the level of Q23L. Analysis of 3-D structure and sequence similarity were used to reveal the presumable factors influencing the enzymatic properties of Q23LG272E, and discussion for the relationship between structure and function of phytase was given.
6-Phytase ; chemistry ; genetics ; metabolism ; Amino Acid Substitution ; Aspergillus fumigatus ; enzymology ; genetics ; Biocatalysis ; Electrophoresis, Polyacrylamide Gel ; Fungal Proteins ; chemistry ; genetics ; metabolism ; Hydrogen-Ion Concentration ; Models, Molecular ; Mutagenesis, Site-Directed ; Mutant Proteins ; genetics ; metabolism ; Mutation ; Pichia ; genetics ; Polymerase Chain Reaction ; Protein Conformation ; Protein Engineering ; methods ; Recombinant Proteins ; metabolism ; Structure-Activity Relationship ; Substrate Specificity

A gene appA encoding a novel phytase was firstly cloned from Hafnia alvei by PCR and sequenced. The gene was consisted of 1335 bp, encoding 444 amino acids. The calculated molecular weight of the mature APPA was about 45.2 kD. The gene appA was expressed in E. coli BL21 (DE3). Recombinant APPA was purified and its enzymatic properties were determined. The optimum pH for the enzyme was 4.5 and the optimum temperature was 60 degrees C. The pH stability of r-APPA is good, the relative phytase activity was above 80% after treated in buffers of pH 2.0-10.0. The specific activity of r-APPA is 356.7 U/mg, and the Km value was 0.49 mmol/L and Vmax of 238 U/mg. The enzyme showed resistance to pepsin and trypsin treatment.
6-Phytase ; biosynthesis ; genetics ; isolation & purification ; Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Hafnia ; enzymology ; genetics ; Hydrogen-Ion Concentration ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Temperature

Fibroblast growth factor 21 (FGF21) is a member of FGF family. It has been demonstrated that FGF21 is an independent, safe and effective regulator of blood glucose levels in vivo. In order to improve the activity of FGF21, we exchanged the beta10-beta12 domain of the human FGF21 with that of the mouse FGF21 to construct a novel FGF21 gene (named hmFGF21), and then subcloned hmFGF21 gene into the SUMO expression vector to create pSUMO-hmFGF21 and transformed it into E. coli Rosetta for expression of the fusion protein SUMO-hmFGF21. Both in vitro and in vivo glucose regulation activity of hmFGF21 was evaluated. The SDS-PAGE result showed that compared with wild-type hFGF21, the soluble expression of hmFGF21 increased about 2-fold. HmFGF21 was more potent in stimulation of glucose uptake in HepG2 cells in vitro. The results of anti-diabetic effect on db/db mice demonstrated that hmFGF21 had better efficacy on controlling the blood glucose of the db/db diabetic animals than wild-type hFGF21. These results suggest that the biological properties of FGF21 are significantly improved by optimization.
Amino Acid Sequence ; Animals ; Blood Glucose ; metabolism ; Cysteine Endopeptidases ; Diabetes Mellitus, Experimental ; blood ; Endopeptidases ; genetics ; Escherichia coli ; Fibroblast Growth Factors ; genetics ; metabolism ; pharmacology ; Genetic Vectors ; Glucose ; metabolism ; Hep G2 Cells ; metabolism ; Humans ; Hypoglycemic Agents ; metabolism ; pharmacology ; Male ; Mice ; Mutation ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Transformation, Genetic