In order to select high quality and suitable Bupleuri Radix varieties in Qingchuan, and establish a new comprehensive method to evaluation the quality of Bupleuri Radix, 12 characters of 14 samples were evaluated by DTOPSIS and grey related degree. The results showed that varieties No. 8 and No. 10 had high quality. DTOPSIS and grey related degree gave the uniformity result, and the biggest difference of value of Ci in DTOPSIS method was 46. 33% , but the biggest difference of the weighting correlation number( r (i)) in grey related degree was only 13.10%. The DTOPSIS combined with grey related degree can evaluate the quality of Bupleuri Radix comprehensively and objectively.
Bupleurum ; chemistry ; China ; Drugs, Chinese Herbal ; chemistry ; supply & distribution ; Quality Control ; Statistics as Topic ; methods

Mesaconitine was incubated with rat liver microsomes in vitro. The metabolites of mesaconitine in rat liver microsomes were identified by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with high resolution power. A typical reaction mixture of 100 mol L-1 Tris-HCI buffer (pH 7.4) containing 0.5 gL-1 microsomal protein and 50 micro molL-1 mesaconitine was prepared. The above reaction mixture was divided into six groups, and the volume of each group was 200 micro L. The incubation mixture was pre-incubated at 37 degrees C for 2 min and the reactions were initiated by adding NADPH generating system. After 90 min incubation at 37 degrees C, 200 micro L of acetonitrile was added to each group to stop the reaction. The metabolites of mesaconitine were investigated by UPLC-MS/MS method. Mesaconitine and 6 metabolites M1-M6 were found in the incubation system. The structures were characterized according to the data from MS/MS spectra and literatures. The metabolic reactions of mesaconitine in rat liver microsomes included the demethylation, deacetylation, dehydrogenation and hydroxylation. The major metabolic pathways of mesaconitine in rat liver microsomes were determined by UPLC-MS/MS on multiple reaction monitoring (MRM) mode combined with specific inhibitors of cytochrome P450 (CYP) isoforms, including alpha-naphthoflavone (CYP1A2), quinine (CYP2D), diethyldithiocarbamate (CYP2E1), ketoconazole (CYP3A) and sulfaphenazole (CYP2C), separately. Mesaconitine was mainly metabolized by CYP3A. CYP2C and CYP2D were also more important CYP isoforms for the metabolism reactions of mesaconitine, but CYP1A2 and CYP2E1 haven't any contribution to MA metabolism in rat liver microsomes.
Aconitine ; analogs & derivatives ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 CYP3A Inhibitors ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Enzyme Inhibitors ; pharmacology ; Ketoconazole ; pharmacology ; Male ; Metabolic Networks and Pathways ; Microsomes, Liver ; enzymology ; metabolism ; Quinine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfaphenazole ; pharmacology ; Tandem Mass Spectrometry

Objective To express the plasminogen activator(Pla) of Yersinia pestis and one of its gene fragments,and to detect their immunological reactivity.Methods The pla gene and its specific gene fragment pla-c were amplified by PCR using the EV76 strain as a template.PCR products were then ligated with the plasmid pET32a (+).The recombinant plasmids pET32a (+)-pla and pET32a (+)-pla-c were subsequently trausformed into E.coli BL21 (DE3).The expressed products were purified by HIS affinity chromatography,and their immunological reactivity was detected by Western blotting.Results The recombinant Pla(52.8 × 103) was expressed as inclusion bodies,and the recombinant Pla-c protein (24.0 × 103) was expressed in the soluble form.These two recombinant proteins reacted with anti-Yersinia pestis EV76 rabbit sera.Conclusions The recombinant Pla and its specific fragments have displayed immunological reactivity,and can be served as an alternative diagnosis method for Yersinia pestis.

Objective To explore the relationship between wild rodent plague and human in wild rodent plague foci of the northwestern area in Yunnan to probe the possible transmission mechanism of wild rodent plague to human. Methods Data of component ratio of rodents and fleas was collected in different areas from 1985 - 1995. Activities and habits of residents regarding the way they keep cats and dogs and parasitic fleas and free fleas indoor were investigated, the dog serum was collected for detecting F1 antibody. Results Eothenomys miletus were main rodents in farmland and shrub, accounting for 48.00% (4753/9902) and 54.50% (4282/7857), Apodemus chevrieri were main rodents in garden, being 50.47% (1332/2639). The component ratio of Neopsylla specialis specialis was 13.31%(229/1720), 12.31%(1678/13 739) and 10.87%(957/8802) respectively in garden, farmland and shrub, higher than in indoor. The component ratio of Frantcpsylla spodix was 39.88% (686/1720), the highest in garden. Thirty-two per cent (32/100) of residents kept cats,in which 63% (20/32) with cat fleas, 68% (68/100) of villages kept dogs, in which 76%(52/68) with fleas. Eighteen parasitic fleas were caught from 43 dogs with a flea index of 0.119 and a rate for fleas of 11.63%, 7 pulex were collected from 17 indoor. Forty-three blood serum samples were obtained from dogs, among which 3 were positive blood serum. Conclusions Residents touch affected animals or media in different situations. The possibility of transmission for wild rodent plague to human exists in loci in a chain of wild rodent plague → fleas or predation → homebred animal plague (cats or dogs) →touching or respiratory → human.

This research use "3414" fertilizer effect experiments to handle zinc, boron and molybdenum trace element fertilizer, determined the dry matter accumulation and content of saikosaponion a and d, to investigate the different ratio of zinc, boron and molybdenum on yield and saikosaponin a, saikosaponin d contents of Bupleurum chinense. Found The suitable ratio of zinc, boron and molybdenum play an active role on dry matter accumulation and distribution, the treatment Zn2B2Mo3 is the best one to promote the dry matter accumulation and transfer to the underground part; in a certain range, only use zinc or molybdenum can promote the yield of B. chinense, the yield of treatment Zn2B2Mo1 is the highest one. According to the results of regression analysis: in accordance with Zn 48.45 g x hm(-2), B 355.05 g x hm(-2), Mo 86.40 g x hm(-2), can obtain the yield with 3313.05 kg x hm(-2); the treatment Zn2BMo2 is most effective to promote the total saikosaponin a and d accumulated, according to the results of regression analysis: in accordance with Zn 36.15 g x hm(-2), B 343.05 g x hm(-2), Mo 106.35 g x hm(-2), the content of total saikosaponin a and d can reach 1.23%. This research first discovered the suitable ratio of zinc, boron and molybdenum can promote the yield and saikosaponin a, saikosaponin d contents on B. chinense.
Boron ; metabolism ; Bupleurum ; metabolism ; Fertilizers ; Molybdenum ; metabolism ; Oleanolic Acid ; analogs & derivatives ; metabolism ; Plant Roots ; metabolism ; Saponins ; metabolism ; Trace Elements ; metabolism ; Zinc ; metabolism

OBJECTIVETo investigate the mechanical changes of the degenerated lumbar disc with finite element analysis.

METHODSA three dimensional finite element model of a human lumbar spine at the L3-L4 disc was established by the software MIMICS and ABAQUS based on computer tomography images. Degeneration was modeled by changes in geometry and material properties. The model was loaded with 0.3 MPa in axial plane. The von Mises stress on the annulus fiber, nucleus pulposus, endplate and facet joints in healthy and degenerated discs was compared.

RESULTCompared with healthy discs, the von Mises stress of disc distributed in the side of annulus fiber, the stress of nucleus pulposus decreased remarkably, the stress of endplate distributed in the posterior part, and the stress of facet joints increased for the degenerated disc.

CONCLUSIONThe finite element models can provide a method of understanding the relationship between biomechanical performance of the disc due to disc degeneration.

Biomechanical Phenomena ; Finite Element Analysis ; Humans ; Intervertebral Disc ; Intervertebral Disc Degeneration ; diagnosis ; Lumbar Vertebrae ; Stress, Mechanical ; Tomography, X-Ray Computed

Resveratrol, isorhapontigenin and pinosylvin, isolated from Gnetum parvifolium, and their analogues have been synthesized and tested for their inhibitory activity of HIV-1. Natural product 12a and analogues (12d, 12e, 12g) display significant inhibitory activity of HIV-1 replication. Among them, compound 12d (trans-3, 4, 5, 4'-tetrahydroxystilbene) exhibits the most potent anti-HIV-1 activity with an IC50 value of 1.84 micromol x L(-1).
Anti-HIV Agents ; isolation & purification ; pharmacology ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gnetum ; chemistry ; HIV-1 ; drug effects ; physiology ; Inhibitory Concentration 50 ; Leukocytes, Mononuclear ; cytology ; virology ; Molecular Structure ; Plants, Medicinal ; chemistry ; Stilbenes ; chemical synthesis ; chemistry ; isolation & purification ; pharmacology ; Virus Replication ; drug effects

OBJECTIVETo screen the mi-RNA expression profile after interferon treatment in cells infected with hepatitis C virus (HCV).

METHODSHuh-7.5.1 cells was infected with HCV by in vitro transcription and cultured with interferon. The mi-RNA microarray was used to measure the mi-RNA expression in the control group, HCV transcription group and interference group. Intra-group differences were analyzed by the 2 ((-delt delt CT)) method.

RESULTSWith mi-RNA expressed in normal Huh-7.5.1 cells as a benchmark, expressions of 13 kinds of mi-RNAs were up-regulated after HCV infection and then down-regulated following interferon treatment; 7 were down-regulated after HCV infection and then up-regulated following interferon treatment.

CONCLUSIONmi-RNA10a, mi-RNA21, mi-RNA149, mi-RNA152 and mi-RNA210 may be related to hepatitis C virus replication and transcription.

Cell Line, Tumor ; Gene Expression Profiling ; Hepacivirus ; genetics ; Humans ; Interferon-alpha ; pharmacology ; Interferons ; pharmacology ; MicroRNAs ; drug effects ; genetics ; RNA, Viral ; genetics ; Transfection

Design space approach is applied in this study to enhance the robustness of first ethanol precipitation process of Codonopsis Radix (Dangshen) by optimizing parameters. Total flavonoid recovery, dry matter removal, and pigment removal were defined as the process critical quality attributes (CQAs). Plackett-Burman designed experiments were carried out to find the critical process parameters (CPPs). Dry matter content of concentrated extract (DMCE), mass ratio of ethanol to concentrated extract (E/C ratio) and concentration of ethanol (CEA) were identified as the CPPs. Box-Behnken designed experiments were performed to establish the quantitative models between CPPs and CQAs. Probability based design space was obtained and verified using Monte-Carlo simulation method. According to the verification results, the robustness of first ethanol precipitation process of Dangshen can be guaranteed by operating within the design space parameters. Recommended normal operation space are as follows: dry matter content of concentrated extract of 45.0% - 48.0%, E/C ratio of 2.48-2.80 g x g(-1), and the concentration of ethanol of 92.0% - 92.7%.
Chemical Precipitation ; Chemistry, Pharmaceutical ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification

Effects of six kinds of Chinese herb extracts, including Folium Crataegi extract, Herba Epimedii extract, Folium Acanthopanacis Senticosi extract, Trifolium pratense L. extract, Folium Ginkgo extract and Radix Puerariae extract, on the activities of CYP450 isozymes (CYP1A2, CYP2C, CYP2E1, CYP2D, CYP3A) in rat hepatic microsomals were studied by using a UPLC-MS/MS (MRM) and cocktail probe substrates method. The results showed that effects of six kinds of Chinese herb extracts on each CYP450 isozyme activity were inhibitory. The IC50 of Folium Crataegi extract for the inhibition of rat microsomal CYP2D activity was only for 4.04 microg x mL(-1), which showed the highest inhibition; Trifolium pratense L. extract had strong inhibitory action to CYP2D, the IC50 value was 5.73 microg x mL(-1); Folium Crataegi extract also had strong inhibitory action on CYP2E1, the IC50 value was 10.91 microg x mL(-1). Furthermore, the IC50 of Folium Ginkgo extract for the inhibition of rat microsomal CYP3A, 2D, 2E1 activities were 45.12, 35.45 and 22.41 microg x mL(-1), respectively, and the IC50 of Folium Acanthopanacis Senticosi extract on the inhibition of rat microsomal CYP2E1 activity was 32.89 microg x mL(-1). In addition, mechanism of inhibition experimental results showed that the inhibiting abilities of Folium Crataegi extract and Radix Puerariae extract on each CYP450 isozyme increased with the increasing of the preincubation time, therefore, the inhibitory effects were a mechanism-based inhibition.
Animals ; Chromatography, High Pressure Liquid ; Crataegus ; chemistry ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP2E1 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Eleutherococcus ; chemistry ; Epimedium ; chemistry ; Ginkgo biloba ; chemistry ; Inhibitory Concentration 50 ; Male ; Microsomes, Liver ; enzymology ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; Trifolium ; chemistry