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1.
China Pharmacy ; (12): 466-470, 2023.
Artículo en Chino | WPRIM | ID: wpr-962493

RESUMEN

OBJECTIVE To analyze the influencing factors for the metabolism of voriconazole in adult patients, and to provide reference for the rational use of voriconazole in clinic. METHODS The clinical data of adult patients admitted in our hospital receiving voriconazole and therapeutic drug monitoring from April 2021 to March 2022 were collected. The trough concentration of voriconazole (c0) and plasma concentration of voriconazole-N-oxide concentration (cN) were determined, and voriconazole-to-voriconazole N-oxide concentration ratio (c0/cN) was calculated. Pearson’s correlation analysis was used to analyze the influencing factors for c0 and c0/cN of voriconazole. Multiple linear regression models were used to analyze the independent influencing factors for c0 and c0/cN of voriconazole. RESULTS The underlying diseases of the patients were mainly pneumonia, kidney disease and leukemia. The detected fungi were mainly Aspergillus, Candida and yeast-like fungi. Voriconazole was mainly administered by intravenous drip, especially in patients who used proton pump inhibitor in combination. The levels of C-reactive protein (CRP), total bilirubin (TBIL), direct bilirubin (DBIL) and indirect bilirubin (IBIL) were positively correlated with c0 of voriconazole, while platelet count and albumin levels were negatively correlated with voriconazole c0. The levels of CRP, TBIL and DBIL were positively correlated with c0/cN, while albumin levels were negatively correlated with c0/cN. Multiple linear regression analysis showed that the independent influencing factors of voriconazole c0 included the levels of CRP and IBIL, route of administration and dose of voriconazole, and the independent influencing factors of voriconazole c0/cN were the levels of CRP and DBIL and age. CONCLUSIONS The levels of CRP and IBIL, route of administration and dose of voriconazole are independent influencing factors of voriconazole c0; the levels of CRP and DBIL and age are independent influencing factors of voriconazole c0/cN. The influence of above indexes on the metabolism of voriconazole should be considered when using voriconazole clinically; and the route of administration and dose of voriconazole should be adjusted reasonably.

2.
China Pharmacy ; (12): 2885-2889, 2021.
Artículo en Chino | WPRIM | ID: wpr-906656

RESUMEN

OBJECTIVE:To investigate the consistency and difference of f luorescence immunochromatographic and liquid chromatography-tandem mass spectrometry (LC-MS/MS)and enzyme multiplied immunoassay technique (EMIT)in the blood concentration monitoring of mycophenolic acid. METHODS :Fluorescence immunochromatography ,LC-MS/MS and EMIT were used to detect the blood concentration of mycophenolic acid in 61 blood samples of children treated with mycophenolate mofetil ester orally at different time points. Kolmogorov-Smirnov method ,Wilcoxon pairing test ,Passing-Bablok regression ,Cusum method,Spearman correlation analysis ,Bland-Altman scatter diagram were adopted for statistical analysis. RESULTS :Blood concentrations of mycophenolic acid ,which were determined by fluorescence immunochromatography ,LC-MS/MS and EMIT , showed non-normal distribution. Passing-Bablok regression analysis showed that regression equation of fluorescence immunochromatography and LC-MS/MS ,fluorescence immunochromatographic method and EMIT were CFI=0.928 3CLC-MS/MS+0.961 7 and CFI=0.880 7CEMIT-0.488 2(FI means fluorescence immunochromatographic ). Spearman correlation analysis showed that the correlation coefficients between fluorescence immunochromatography and LC-MS/MS ,fluorescence immunochromatography and EMIT were 0.968 and 0.929, respectively (P<0.000 1). Bland Altman scatter plot analysis showed that 3.28% of the 358341451@qq.com difference between fluorescence immunochromatography and LC-MS/MS was outside the consistency limit (±1.96SD), and 1.64% of the difference between fluorescence immuno- chromatography and EMIT was outside the consistency limit (± 1.96SD). Wilcoxon pairing test showed that the results of fluorescence immunochromatography were higher than those of LC-MS/MS (Z=3.76,P=0.000 2)and lower than those of EMIT (Z=-5.96,P<0.000 1). CONCLUSIONS :Fluorescence immunochromatography shows good consistency and correlation with LC-MS/MS and EMIT ;the blood concentrations of mycophenolic acid detected by fluorescence immunochromatography were higher than those by LC-MS/MS and lower than those by EMIT . It can be used for bedside rapid detection. When using the test results of different methods for clinical medication ,the differences of test methods need to be considered.

3.
China Pharmacy ; (12): 2946-2951, 2019.
Artículo en Chino | WPRIM | ID: wpr-817474

RESUMEN

OBJECTIVE: To establish the method for simultaneous determination of clopidogrel (CLP), its intermediate metabolite (2-O-CLP), inactive metabolite (CLPCA) and active metabolite (CLPTM) in human plasma. METHODS: Totally 90 patients diagnosed as stroke were selected from the First Affiliated Hospital of Army Medical University. They were given one CLP tablet (75 mg/tablet) orally on an empty stomach in the morning. Blood samples were collected 2 h after taking the tablet. CLPTM- D was formed by derivation of CLPTM with 2-bromo-3’-methoxyacetophenone and extracted by precipitation of acetonitrile protein together with the other three substances to be measured. LC-MS/MS method was adopted. The determination was performed on Agilent poroshell 120 EC-C18 column with mobile phase consisted of acetonitrile (0.1% formic acid) and water (0.1% formic acid) (90 ∶ 10, V/V). The quantitation analysis was performed using multiple reaction monitoring at the specific ion transitions of m/z 308.1→198.1 (CLPCA), 322.3→212.0 (CLP), 338.3→155.0 (2-O-CLP), 504.4→354.1 (CLPTM-D) and 264.0→154.1 (ticlopidine, internal standard), respectively. RESULTS: The retention time of CLPCA, CLP, 2-O-CLP, CLPTM-D and internal standard were 2.01, 3.32, 2.83, 2.68, 1.87 min, respectively. The linear range of CLPCA, CLP, 2-O-CLP and CLPTM-D were 100-10 000, 0.2-20, 0.3-30, 0.5-50 ng/mL (all r≥0.999 5). The intra-day and inter-day RSD were all less than 9.5% (n=5). Accuracy ranged from 93.5%-98.9% (n=5), and extraction recovery was from 85.4% to 95.9% (n=5). The matrix effect ranged from 2.7%-6.2% (n=5). In stability tests (storing at -80 ℃ for 3 months, 3 freeze-thaw cycles, storing at 4 ℃ for 8 h), RE of CLP, CLPCA and CLPTM-D were all lower than 10.0% (n=5). CONCLUSIONS: Established LC-MS/MS method has the advantages of high specificity, accuracy and reliability, and can be used to detect the concentration of CLP and its three metabolites in human plasma.

4.
Chongqing Medicine ; (36): 2617-2619, 2016.
Artículo en Chino | WPRIM | ID: wpr-495419

RESUMEN

Objective To study the biofilm formation ability and related gene distribution of Staphylococcus (S .) aureus iso‐lated from urinary tract infections to provide the theoretical basis for the prevention and treatment of clinical infection .Methods The minimal inhibitory concentration was detected using the agar double dilution method .The bacterial adhesion ability was deter‐mined by flat colony counting method .The biofilm formation ability was analyzed by the 96‐well crystal violet staining method .The biofilm‐associated genes were detected by PCR amplification .Results Eleven clinical strains of S .aureus were high resistant to pen‐icillin and erythromycin ,whereas were all sensitive to vancomycin and nitrofurantoin .All the isolates had a strong ability of adhe‐sion ,but the biofilm formation ability was weak .Among them ,the icaAD and icaBC genes were amplified in 10 S .aureus isolates . Conclusion The adhesion ability and biofilm formation ability of S .aureus isolated from urinary tract infections have the strain differences ,and ica is an important gene of S .aureus biofilm formation .

5.
China Pharmacy ; (12): 2765-2767, 2016.
Artículo en Chino | WPRIM | ID: wpr-504539

RESUMEN

OBJECTIVE:To investigate synergistic effect of carbapenems combined with fosfomycin(FOS)on carbapenems-re-sistant Pseudomonas aeruginosa isolates from urinary tract infections in vitro. METHODS:The minimum inhibitory concentration was detected using agar double dilution method. The fractional inhibitory concentration index was determined by checkerboard meth-od. The effect of carbapenems combined with FOS on biofilm of P. aeruginosa isolates was determined using 96 crystal violet stain-ing. RESULTS:12 strains of carbapenem-resistant P. aeruginosa isolates were highly sensitive to FOS and amikacin,and were com-pletely resistant to imipenem and meropenem. The combination of imipenem with FOS could induce a synergistic effect on 4 strains (33.3%);meropenem combined with FOS could induce a synergistic effect on 5 strains(41.7%);no antagonistic effect of carbap-enems combined with FOS appeared. FOS combined with carbapenems could inhibit the biofilm of carbapenems-resistant P. aerugi-nosa(P<0.05 or P<0.01). CONCLUSIONS:The combination of carbapenems with FOS possesses in vitro synergistic antibacteri-al effect on part of carbapenems-resistant P. aeruginosa isolates,the mechanism of which may be associated with inhibiting the bio-film.

6.
Chinese Journal of Burns ; (6): 285-289, 2015.
Artículo en Chino | WPRIM | ID: wpr-327386

RESUMEN

<p><b>OBJECTIVE</b>To study the effects of inhibitory peptide of Staphylococcus epidermidis (SE) biofilm (briefly referred to as inhibitory peptide) on adhesion and biofilm formation of SE at early stage.</p><p><b>METHODS</b>By using peptide synthesizer, the inhibitory peptide was synthesized with purity of 96.8% and relative molecular mass of 874.4. (1) Solution of SE ATCC 35984 (the same below) was cultivated with inhibitory peptide in the final concentrations of 1-256 µg/mL, and the M-H broth without bacteria solution was used as blank control. The MIC of the inhibitory peptide against SE was determined (n=3). (2) Solution of SE was cultivated with trypticase soy broth (TSB) culture solution containing inhibitory peptide in the final concentrations of 16, 32, 64, 128, and 256 µg/mL (set as inhibitory peptide groups in corresponding concentration), and solution of SE being cultivated with TSB culture medium was used as negative control group. Growth of SE was observed every one hour from immediately after cultivation (denoted as absorbance value), and the growth curve of SE during the 24 hours of cultivation was drawn, with 3 samples in each group at each time point. (3) Solution of SE was cultivated with TSB culture solution containing inhibitory peptide in the final concentrations of 16, 32, 64, 128, and 256 µg/mL (set as inhibitory peptide groups in corresponding concentration), and solution of SE being cultivated with TSB culture medium was used as negative control group. Adhesive property of SE was observed after cultivation for 4 hours (denoted as absorbance value, n=10); biofilm formation of SE was observed after cultivation for 20 hours (denoted as absorbance value, n=10). (4) Solution of SE was cultivated with TSB culture solution containing inhibitory peptide in the final concentration of 128 µg/mL (set as 128 µg/mL inhibitory peptide group), and solution of SE being cultivated with TSB culture medium was used as negative control group. Adhesive property of SE and its biofilm formation were observed with confocal laser scanning microscope (CLSM), and the sample numbers were both 3. Data were processed with one-way analysis of variance, LSD test, and Dunnett T3 test.</p><p><b>RESULTS</b>(1) The MIC of inhibitory peptide against SE exceeded 256 µg/mL. (2) There was no significant difference in the growth curve of SE between inhibitory peptide groups in different concentrations and negative control group. (3) After 4 hours of cultivation, the absorbance values of adhesive property of SE in 256, 128, 64, and 32 µg/mL inhibitory peptide groups were respectively 0.20 ± 0.04, 0.27 ± 0.03, 0.35 ± 0.04, and 0.40 ± 0.04, which were significantly lower than the absorbance value in negative control group (0.53 ± 0.10, P<0.05 or P<0.01); the absorbance value of adhesive property of SE in 16 µg/mL inhibitory peptide group was 0.47 ± 0.09, which was close to the absorbance value in negative control group (P>0.05). After 20 hours of cultivation, the absorbance values of biofilm formation of SE in 256, 128, and 64 µg/mL inhibitory peptide groups were respectively 0.49 ± 0.10, 0.68 ± 0.06, and 0.93 ± 0.13, which were significantly less than the absorbance value in negative control group (1.21 ± 0.18, P<0.05 or P<0.01); the absorbance values of biofilm formation in 32 and 16 µg/mL inhibitory peptide groups were respectively 1.18 ± 0.22 and 1.15 ± 0.26, which were close to the absorbance value in negative control group (with P values above 0.05). (4) CLSM showed that more adhering bacteria and compact structure of biofilm were observed in negative control group, but less adhering bacteria and loose structure of biofilm were observed in 128 µg/mL inhibitory peptide group.</p><p><b>CONCLUSIONS</b>The inhibitory peptide can inhibit adhesion and biofilm formation of SE at early stage, but its structure still needs to be further modified.</p>


Asunto(s)
Humanos , Adhesión Bacteriana , Biopelículas , Microscopía Confocal , Péptidos , Staphylococcus epidermidis , Genética , Metabolismo , Fisiología
7.
Chongqing Medicine ; (36): 1931-1933, 2015.
Artículo en Chino | WPRIM | ID: wpr-468185

RESUMEN

Objective To analyze the distribution and drug resistance situation of pathogens in the respiratory department during the recent 9 years to provide the basis for rational use of antibacterial drugs in clinic .Methods All pathogens isolated from the respiratory depeartment from January 2003 to December 2011 and the drug susceptibility test results were retrospectively and statistically analyzed .Results A total of 5 714 strains of pathogenic bacteria were isolated ,which mainly distributed in the sputum (90 .1% ) ,excrement (4 .2% ) and urine (3 .6% );among them ,2 943 strains (51 .5% ) were Gram‐negative bacteria ,596 strains (10 .4% ) were Gram‐positive bacteria and 2 175 strains (38 .1% ) were fungi .The top six of isolated bacteria were Candida albi‐cans ,Pseudomonas aeruginosa ,Acinetobacter baumannii ,Klebsiella pneumoniae ,Candida tropicalis and Escherichia coli .The isola‐tion rates of A .baumannii and C .albicans were increased year by year ,while the isolation rate of E .coli was decreased .A .baumannii and P .aeruginosa had a high resistant to all antibacterial drugs ,whereas the resistant rate of A .baumannii was increased year by year and that of P .aeruginosa showed some fluctuation .K .pneumoniae had a high susceptibility to imipenem and meropenem ,and the sensitivity to other antimicrobial agents had a gradually increasing tendency .The sensitive rate of C .albicans to amphotericin B was almost 100% ,and they had a high susceptible to other antifungal agents .Conclusion Drug resistance of the pathogens is com‐mon in the respiratory department .It is of importance to emphasize the pathologic examination ,carry out the surveillance of drug re‐sistance of pathogenic bacteria ,and use the antibacterial drugs rationally in clinical anti‐infective therapy .

8.
Military Medical Sciences ; (12): 694-697, 2015.
Artículo en Chino | WPRIM | ID: wpr-479179

RESUMEN

Objective To investigate the regulation of swimming motility by H-NS in Vibrio parahaemolyticus(VP). Methods VP was inoculated into the semi-solid swimming agar plate containing 1% Oxoid tryptone, 2% NaCl, 0.5%Difco Noble Agar, and 0.1% arabinose followed by incubation at 37℃ for 4.5 h before the diameters of bacterial lawns were measured.Total RNAs were extracted from the wild-type (WT) strains and the hns null mutant (Δhns), and the quantitative real-time( RT)-PCR( qRT-PCR) was carried out to calculate the transcriptional variation of flaA between WT andΔhns strains.The entire promoter DNA region of flaA was amplified and cloned into the lacZ fusion vector pHRP309 containing a promoterless lacZ gene. The recombinant lacZ reporter plasmid was transformed into WT and Δhns, respectively, to measure the β-galactosidase activities in cellular extracts using the β-galactosidase enzyme assay system. Results and Conclusion The phenotype results showed that swimming motility of VP was enhanced by H-NS.The qRT-PCR and LacZ fusion results indicated that the transcription of flaA was positively regulated by H-NS.Collectively, H-NS promotes the swimming motility of VP, at least partly, by activating the transcription of flaA.

9.
Artículo en Chino | WPRIM | ID: wpr-333624

RESUMEN

<p><b>OBJECTIVE</b>To study the effect of DNase I on biofilm formation of Staphylococcus aureus.</p><p><b>METHODS</b>The growth curve of S. aureus was detected using a spectrophotometer. The adhesion of S. aureus was analyzed using flat colony counting method, and the biofilm formation was assayed using the 96-well crystal violet staining method.</p><p><b>RESULTS</b>Exposure to different concentrations of DNase I did not obviously affect the growth of S. aureus but significantly inhibit the formation of bacterial biofilms in a dose-dependent manner. DNase I inhibited the adhesion of S. aureus at different growth stages. When combined with antibiotics, DNase I resulted in a signi?cant decrease in the established bio?lm biomass compared to antibiotics or DNase I used alone.</p><p><b>CONCLUSION</b>DNase I can effectively inhibit biofilm formation of S. aureus and enhance the inhibitory effect of antibiotics against S. aureus biofilms.</p>


Asunto(s)
Antibacterianos , Biopelículas , Desoxirribonucleasa I , Química , Staphylococcus aureus
11.
China Pharmacy ; (12)2007.
Artículo en Chino | WPRIM | ID: wpr-533057

RESUMEN

OBJECTIVE:To investigate the effect of subinhibitory concentration of erythromycin on adhesion of clinical isolated Staphylococcus(S.) epidermidis.METHODS:The subinhibitory concentration of erythromycin was determined based on the susceptibility test,and the representative strain Se.015 was treated with subinhibitory concentration of erythromycin.The optical density value determined by microtiter-plate assay was used to evaluate the effect of erythromycin on the adhesion of the representative strain Se.015,and electron microcopy was employed to observe the adhesion of Se.015 in samples with blank solvent served as control.RESULTS:Compared with control group,erythromycin(4 mg?L-1) group showed significantly higher optical density value(P

12.
Artículo en Chino | WPRIM | ID: wpr-566110

RESUMEN

Objective To investigate the correlation of biofilm forming ability with biofilm associated gene of the clinically isolated Staphylococcus aureus for providing basis for the further studies of the mechanisms of biofilm formation.Methods Safranine staining was conducted for the detection of the forming ability of biofilm in 96-well plates.icaAD,icaBC,sar,agr and sigB was amplified by PCR.Results Formation of biofilm could be found macroscopically in 17 out of 27 strains,especially to X387 and X409.icaAD and icaBC were amplified in 22 isolates and sar,agr and sigB in all 27 S.aureus strains.Conclusion ica operon is the key gene for biofilm formation but cooperative action of other genes is needed during the process of biofilm formation.

13.
Artículo en Chino | WPRIM | ID: wpr-566670

RESUMEN

Objective To study the macrolide-resistance,ability of biofilm formation and icaA-genetypes of 68 stains of clinically isolated Staphylococcus epidermidis(S.epidermidis) so as to explore the efficacy of macrolide to prevent biofilm-associated infections caused by S.epidermidis.Methods Minimum inhibitory concentration(MIC) were detected by agar-plate dilution method,and microtiter-plate assay was used to investigate the ability of biofilm formation,and icaA genetype was identified by PCR.Results High macrolide-resistant rate(88.2%) was showed for clinical isolated S.epidermidis,and biofilms for macrolide-resistant strains were significantly stronger than that of macrolide-sensitive strains,but there was no dependability found between these two for icaA positive rate.Conclusion Frequently-used macrolide such as erythromycin,azithromycin,and clarithromycin may incompetence to prevent biofilm-associated infections caused by S.epidermidis.

14.
China Pharmacy ; (12)2001.
Artículo en Chino | WPRIM | ID: wpr-527548

RESUMEN

OBJECTIVE:To study the?-lactamase production in multi-drug resistant acinetobacter baumannii isolated in clinic.METHODS:Susceptivity to antibiotics of the bacteria was measured by K-B and agar diffusion methods.The?-lac-tamase production of acinetobacter baumannii was examined by nitrocefin disc test,then their plasmid and chromosome ex-tracted as templete,the genes coded the?-lactamase were amplified by PCR with commercial kits.Furthermore,the sequence and homology of PCR products were analyzed.RESULTS:Total12acinetobacter baumannii in this study were?-lacta-mases-producing strains with a high resistance to cephalosporin.However,it is sensitive to carbapenem-antibiotics and cephalosporin with?-lactamase inhibitors,and6strains of them were confirmed that there were?-lactamases AmpC gene on plasmid by PCR amplification and sequence analysis.CONCLUSIONS:The?-lactamases AmpC mediated by plasmid would be main factor in the high resistance to cephalosporin of acinetobacter baumannii isolated clinically.

15.
China Pharmacy ; (12)2001.
Artículo en Chino | WPRIM | ID: wpr-527710

RESUMEN

OBJECTIVE:To construct the homologous recombination plasmid of sigB gene from staphylococcus aureus so as to provide the basis for further study of the formation mechanism of its biofilm.METHODS:The target fragment“upstream sequence of sigB,kana-resistance gene,and downstream sequence of sigB”were amplified by PCR from staphylococcus aureus DNA and pEGFP-N2plasmid,respectively,in which the terminals of3interlinked fragments were complementary,and re-stricted enzyme EcoRⅠand XhoⅠsite existed in the sequence terminal,respectively.The fragments were ligated with fusion PCR in turn.Then target fragment was recombinated into plasmid pBluescriptⅡSK(+)according to the instruction manual of commercial kits.After both recombination of pBluescriptⅡSK(+)and vector plasmid pGEM-7zf(+),restricted enzyme SacⅠand XhoⅠ,the homologous recombination plasmid pGEM-7zf(+)-sigB was finally made via T 4 ligation enzyme.RE-SULTS:The target fragments were amplified by PCR and were confirmed with the agarose gel electrophoresis and sequencing.CONCLUSION:The homologous recombination plasmid of sigB gene from staphylococcus aureus is constructed successfully.

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