OBJECTIVE: To determine Ginsenoside Rg1,Ginsenoside Re and Ginsenoside Rb1 in Yixin fumai granula by HPLC.METHODS: Samples were determined on a Diamond C18(150 mm?4.6 mm,5 ?m).The mobile phase consisted of accetonitrile-water by gradient elution with flow rate at 1 mL?min-1,UV detection wavelength at 203 nm,column temperature at 25 ℃ and sample size at 10 ?L.RESULTS: The linear ranges of Ginsenoside Rg1,Ginsenoside Re and Ginsenoside Rb1 were 0.645～6.450 ?g(r=0.999 8),0.54～5.40 ?g(r=0.999 7) and 0.605～6.050 ?g(r=0.999 9),respectively;the average recoveries were 100.59%(RSD=2.03%),98.70%(RSD=1.46%)and 98.99%(RSD=1.19%),respectively.CONCLUSION: The method is sensitive,simple and accurate,and it can be used for the quality control of Yixin fumai granula.

Humans ; Lymphoma, Large-Cell, Anaplastic ; diagnosis ; enzymology ; therapy ; Receptor Protein-Tyrosine Kinases

Anticonvulsants ; adverse effects ; Humans ; Seizures ; chemically induced

Adult ; Ammonia ; poisoning ; Female ; Humans ; Microscopy, Electron ; Poisoning ; complications ; Respiratory Function Tests ; Respiratory System ; drug effects ; pathology ; ultrastructure ; Time Factors

Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.

Survivin is a protein that inhibits apoptosis and regulates cell division.The cDNA sequence of survivin was amplified by RT-PCR and sub-cloned into the prokaryotic expression vector pET-21b(+),followed by transformation into E.coli strain BL21(DE3) and induction with IPTG.The recombinant survivin protein fusing with 6?His tag was expressed in E.coli in the form of inclusion body at the expression level over 60% of the total cell protein.Results of Western blotting showed that recombinant survivin reacted specifically with anti-human survivin antibody.After gel filtration,the recombinant protein reached the purity over 95%,which facilitate the study of diagnosing and inhibitor agents targeting survivin.

To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.

Objective To construct the serine protease gene(sprE)mutant and to study the pathogenicity of sprE gene of Enterococcus faecalis.Methods Recombinant suicide vector pCQ001 of Enterococcus faecalis with pTX4577,was constructed.Then,created isogenic sprE-deficient mu- tant(*sprE)by allelic replacement was constructed.Moreover,the growth ability and the virulence of the mutant were compared with those of the wide type in vitro and in vivo respectively.A mouse peritonitis model and a rabbit endocarditis model were utilized in the study.Results The *sprE was selected by kanamycin and identified by polymerase chain reaction(PCR),pulsed field gel electropho- resis(PFGE)and Southern blot.The evidences showed that the sprE gene had a major role in helping bacteria to resist the elevated temperature and oxidative stress.The virulence of mutant decreased af- ter sprE gene was knocked out.Conclusions The *sprE of Enterococcus faecalis is constructed suc- cessfully,sprE gene is important in the pathogenesis of Enterococcus faecalis,which probably is a major virulence factor of Enterococcus faecali.

Objective To investigate the reliability of a novel rating scale, unified multiple system atrophy rating scale, section Ⅰ(UMSARS-Ⅰ) in the evaluation of illness severity in patients with multiple system atrophy (MSA). Methods A retrospective analysis and a prospective follow-up study were conducted by using UMSARS-Ⅰ in 46 patients with MSA, and the Schwab and England scale was employed and illness severity was graded. The reliability, validity and sensitivity to change of UMSARS-Ⅰ in evaluating the illness severity of MSA were estimated. Results UMSARS-Ⅰ enjoyed high internal consistency (standard Crohnbach's ?=0.88) and sound content, criterion-related, construct and discriminant validity in the evaluation of illness severity of MSA, and a moderate sensiti-vity to change was found(effect size=0.61). Conclusion UMSARS-Ⅰ is a reliable and multidimensional semi-quantitative scale in the measurement of severity and progression of impairment in MSA.

Objective To examine the relationship between plasma homocysteine level and status of congestive heart failure. Methods Plasma homocysteine level was determined in 106 patients with congestive heart failure(CHF).Among them,40 patients were diagnosed as having recent onset of CHF(group 1) and the remaining 66 were receiving conventional treatment(group 2).Thirty healthy subjects were served as a control group. Results(The plasma) homocysteine levels in group 1,group 2 and the control group were(14.87?5.22),(13.25?5.45) and((7.52)?1.73) ?mol/L,respectively.The plasma homocysteine level was significantly higher in group 1 and group 2 than in the control group(P