Objective To study the content of taxol in different parts of Taxus Yunnanensis and the effects of different growth time on taxol content. Method A HPLC method was developed for the detetmination of taxol content in bark, needle, twig, root from Taxus Yunnanensi, and the effects of different growth time. Results Taxol content in bark was the highest, that in root was the lowest and that in needle and twig were higher. Taxol content was higher with the growth time in needle and twig of Taxus Yunnanensi. Conclution It was significant to take taxol from needle and twig of Taxus Yunnanensis which is more than five years growth time.

Objective To control the quality of Herba Leonuri by studying the content of stachydrine hydrochloride in different parts of it. Method A HPLC method was developed for the content detetmination of stachydrine hydrochloride in flower, stem, leafage from Herba Leonuri. Spherisorb SCX colum was used with mobile phase of 20 mmol/L sodium dihydrogen phosphate (containing 0.04% triethylamine and 0.15% phosphoric acid). The colum temperature was 25 ℃, the detective wavelength was 192 nm. Results Stachydrine hydrochloride content was the highest in leafage and the lowest in stem. Conclusion To ensure the quality of Herba Leonuri, it is significant to choose medical material with more leafage.

Adult ; Dust ; analysis ; Female ; Humans ; Male ; Middle Aged ; Neural Networks (Computer) ; Occupational Exposure ; Silicosis ; diagnosis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

Two patients were admitted to the hospital for 2-month history of pruritic eruptions on the forehead and 2-week history of pruritie eruptions on the leg, respectively. Both patients had a history of pet contact. Topical application of glucocorticoids did not work well. Dermatological examination revealed a patch measuring 5 cm ×6 cm on the forehead of one patient and a patch measuring 2 cm × 3 cm on the leg of the other patient. Both patches were surrounded by red papules and scaling. Microscopic examination of skin scales revealed hyphae and chain-like spores, and culture of skin scales grew Microsporum gyeum. Both the isolates of Microsporum gyeum were sensitive to ketoconazole, miconazole, bifonazole, terbinafine, and voriconazole. Both patients were healed after treatment with oral terbinafine and topical ciclopirox olamine.

Objective To investigate the in vitro susceptibility of 16 strains of Exophiala dermatitidis to 6 commonly used antifungal agents. Methods The Glinical and Laboratory Standards Institute (CLSI)M27-A2 protocol was carried out to determine the MIGs of terbinafine (TRB), itraconazole (ITC), amphotericin B (AMB), fluconazole (FLC), voriconazole (VRC), and caspofungin (GAS) to 16 strains of E. dermatitidis, and E-test was performed to determine those of VRG, ITC and AMB. Besides, the minimal fungicidal concentrations (MFGs) of the above antifungal agents to the 16 strains of E. dermatitidis were further assessed.The activity of TRB in combination with ITC and VRG against E. dermatitidis was also estimated. Results The MIC ranges of TRB, VRC, ITC, AMB, FLC, and CAS were 0.125 - 0.25 mg/L, 0.25 - 0.5 mg/L, 2.0 mg/L,2.0 mg/L, 16 - 32 mg/L and 16 - 64 mg/L respectively as shown by M27-A2 microdilution assay, while the MIC ranges of VRG, ITG and AMB, as determined by E-test, were 0.032 - 0.094 mg/L, 0.047 - 0.5 mg/L and 0.125 - 3.0 mg/L, respectively. The MFC ranges of TRB, VRC, ITG, AMB and FLG were 0.125 - 0.5 mg/L,0.25 - 0.5 mg/L, 2.0 - 4.0 mg/L, 2.0 - 4.0 mg/L and 16 - 64 mg/L, respectively. No synergism in the acitivity against E. dermatitidis was observed for the combination of TBR with ITC or VRC. Conclusion E. dermatitidis is susceptible to TRB, ITC, AMB, and VRC, but less sensitive to both FLC and GAS.

Objective To investigate the molecular mechanisms of cross-resistance to azoles in a clinical isolate of Aspergillus fumigatus. Methods A. fumigatus was isolated from a patient with invasive aspergillosis.Clinical Laboratory Standard Institute M38-A2 broth microdilution method and E-test method were used to determine the minimum inhibitory concentrations (MICs) or minimum effective concentration (MEC) of itraconazole, voriconazole, amphotericin B, posaconazole and caspofungin for the A. fumigatus isolate. DNA was extracted from the isolate and subjected to the amplification of cyp51A gene encoding the target enzyme of azole antifungal agents followed by sequence analysis. Results The broth microdilution test showed that the MEC of caspofungin was 0.5 mg/L, and MICs of itraconazole, voriconazole and amphotericin B were ≥ 16 mg/L,8 mg/L and 1 mg/L, respectively, for this isolate; while E-test assay revealed that the MICs of caspofungin,itraconazole, voriconazole, amphotericin B and posaconazole were 0.047 mg/L, ≥32 mg/L,≥32 mg/L, 12 mg/L and ≥32 mg/L, respectively. Sequence analysis showed an insertion of a 34-bp tandem sequence in the promoter region of the cyp51A gene as well as a T364A point mutation causing the substitution of leucine 98 (L98H). In addition, there were some other mutations in the cyp51A gene of this isolate, such as A137T,G585A, C814A, G836C, T991C and A1350G, which could result in corresponding amino acid substitutions.Conclusions An A. fumigatus strain with cross-resistance to azole antifungal agents is isolated. There is an insertion of a 34-bp tandem sequence into the promoter region as well as a T364A point mutation in the cyp51A gene, which contribute to the cross resistance to azole antifungal agents including itraconazole, voriconazole,and posaconazole. In addition, other mutations causing amino acid substitutions have also been detected in the cyp51 A gene of this isolate.

Objective To explore the clinical diagnosis,medication,curative effect,and prognosis of lymhocyte subsets and humoral immunity in children with Kawasaki disease(KD)complicated with coronary artery dilatation.Methods One hundred and seventy-one children(age of 4 months to 9 years)from Jun.2000 to Dec.2007 with KD in Capital Institute of Pediatrics were taken as research objective,65 healthy children(age of 5 months to 7 years)were taken as controls,the difference of the level variation of lymhocyte subsets and humoral immunity was detected and analyzed between them.The flow cytometry and scattering turbidimetry method were used to detect the numbers of T cell subsets [CD3+,CD3+CD4+,CD3+CD8+,CD19+,natural killer(NK)cell] and levels of immune globulin(IgG,IgA,IgM)of the peripheral blood of 171 patients,compare them with those of the control group,and compare them between patients with and without coronary artery dilatation.Results Compared with control group,in the acute period of KD the levels of CD3+,CD3+CD4+ decreased significantly(t=0.01,0.02 Pa0.05);of all the 171 children patient,53 cases were coronary artery expansion patients.Compared with non-expansion patients,levels of CD4+/CD8+,CD3+CD8+,IgG of coronary artery expansion patients were significant different and had statistical difference(t=0.02,0.04,0.03 Pa

Objective:To investigate the efficiency of Agrobacterium tumefaciensmediated transfor-mation of Aspergillus fumigatus by using pyrG as a recessive selectable marker.Methods: FAP1 and SHO1 genes target sequences,composed of a selectable marker pyrG and the flanking sequences of the FAP1 and the SHO1 genes,were cloned into a binary plasmid pDHt/sk,respectively.The produced plasmids were transformed into A.tumefaciens.The A.tumefaciens and uracil auxotroph A.fumigatus were cocultured in induction medium without uricil and uridine at 24 ℃ for 48 h.To inhibit growth of A.tumefaciens and to select transformants,the cultures were transferred to 37 ℃ and incubated for another 48 h.Results: In this study,A.tumefaciens-medidated transformation of A.fumigatus produced high homologous recombination rates,which was 44%(7 of 16) for FAP1 and 35%(7 of 20) for SHO1.Conclusion: Our study showed that A.tumefaciens-medidated transformation by using pyrG as a recessive selectable marker is an efficient tool for target gene deletion of A.fumigatus.

A rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometric (RRLC-Q-TOF-MS) method was established and optimized for the analysis of pharmacokinetic behavior of ginsenoside Rb2 in rats by intravenous injection administration.The metabolism of ginsenosides Rb2 in vivo rat was also explored.In the experiment,Agilent SB C18 column was selected for the sample separation with 0.1％ aqueous formic acid solution as mobile phase (A) and acetonitrile as mobile phase (B) at a flow rate of 0.2 mL/min,and the injection volume was set to 5 μL.Q-TOF-MS was carried out in electron pray ionization (ESI) negative ion mode.The limit of quantification (LOQ,S/N =10) and limit of detection (LOD,S/N=3) were 0.10 and 0.08 μg/mL,respectively,and the linear range was 0.1-1.26 μg/mL.The experiment results showed that the concentration-time profile of ginsenoside Rb2 conformed to a two-compartment pharmacokinetic model after intravenous administration for rats.The mean plasma elimination half-lives were (23.58±1.10) min (t1/2α),(1306.55±147.23) min (t1/2β) for Rb2.By analyzing the urine of rats after intravenous administration and the fecal samples after oral administration of ginsenoside Rb2,it was found that the metabolites were M6,M2 (CY),F2,and C-K.

to common antifungal drugs.