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

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.

Anticonvulsants ; adverse effects ; Humans ; Seizures ; chemically induced

The contents of coumarins in the sulfur fumigated Angelicae Dahuricae Radix (Baizhi, ADR) were reduced significantly. To achieve the quality control of ADR, the qualitative identification of sulfur fumigated ADR and quantitative model of imperatorin content should be established. The near-infrared (NIR) spectrograms of non-sulfur and sulfur fumigated ADR were collected by NIR diffuse reflectance spectroscopy technology and pretreated by the method of first derivative derivation and vector normalization. The Ward's Algorithm method was used for the cluster analysis. The non-sulfur and sulfur fumigated ADR can be quickly identified in the range of 8,806. 0-3 811.0 cm(-1) based on the cluster analysis. The NIR quantitative model of imperatorin was established by the contents of imperatorin determined by HPLC in combination with partial least squares regression analysis. According to the calibration model established in this study, correlation coefficients (R2), the root-mean-square error of cross-validation (RMSECV), and the root-mean-square error of prediction (RMSEP) for imperatorin were 0.982 8, 0.006 8, 0.011 8, respectively. The quantitative model of imperatorin can be applied to determine the content of imperatorin in ADR accurately.
Angelica ; chemistry ; classification ; China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Spectroscopy, Near-Infrared ; methods ; Sulfur ; chemistry

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

OBJECTIVETo observe the therapeutic efficacy of Dangfei Liganning Tablet (DLT) in the treatment of antipsychotics induced mild hepatic damage.

METHODSTotally 80 mental inpatients with antipsychotics induced mild liver injury were randomly assigned to two groups, the treatment group (40 cases) and the control group (40 cases). Patients in the treatment group took DLT, two tablets each time, three times per day, while those in the control group took Liver-protecting Tablet (LT), four tablets each time, three times per day. The treatment course was 4 weeks for all. Changes of glutamic-pyruvic transaminase (ALT) and glutamic-oxalacetic transaminase (AST) were observed before treatment, week 1, 2, and 4 after treatment. The therapeutic efficacy was compared between the two groups.

RESULTSCompared with the former time point, ALT and AST gradually decreased in the two groups at week 1, 2, and 4 (P <0. 05). The cured rate was 72. 5% and the total effective rate was 97. 5% in the treatment group. They were 62. 5% and 90. 0% respectively in the control group. There was no statistical difference in the two indices between the two group (P >0.05). No obvious adverse reaction occurred in the two groups.

CONCLUSIONDLT could treat antipsychotics induced mild hepatic damage in a safe and effective way.

Alanine Transaminase ; metabolism ; Antipsychotic Agents ; adverse effects ; Chemical and Drug Induced Liver Injury ; drug therapy ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Liver ; metabolism ; Protective Agents ; administration & dosage ; therapeutic use ; Tablets ; therapeutic use

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.