Owing to the fact that luteolin has antibacterial activity against Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA), its specific mechanism in MRSA is worthy of investigation, which is the focus of this study. Initially, the collected S. aureus strains were treated with luteolin. Then, the minimum inhibitory concentration (MIC) of luteolin against the S. aureus strains was measured by the broth microdilution. The growth curves, biofilm formation, and cytotoxicity of treated S. aureus were detected using a microplate reader. The live and dead bacteria were evaluated using confocal laser scanning microscopy, the bacterial morphology was observed using scanning electron microscopy, and the S. aureus colony-forming unit (CFU) numbers were assessed. The levels of alpha hemolysin (α-hemolysin), delta hemolysin (δ-hemolysin), and hlaA were detected via western blot and RT-PCR. The mortality of mouse model with S. aureus systemic infection was analyzed, and the levels of IL-6, IL-8, IL-10, and TNF-α were quantitated using ELISA. Concretely, the MIC of luteolin against MRSA N315 was 64 μg/mL. Luteolin at 16 μg/mL did not affect the growth of MRSA N315, but inhibited the biofilm formation and CFU, and promoted the morphological changes and death of MRSA N315. Luteolin decreased the cytotoxicity and the levels of α-hemolysin, δ-hemolysin, and hlaA in MRSA N315, elevated MRSA-reduced mice survival rate, and differentially modulated the inflammatory cytokine levels in MRSA-infected mice. Collectively, luteolin inhibits biofilm formation and cytotoxicity of MRSA via blocking the bacterial toxin synthesis.

Background and Objective: Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen resistant to a variety of antibiotics. This study aimed to investigate the in vitro antibacterial activity of luteolin combined with vancomycin against MRSA. Materials and Methods: Ten clinical MRSA isolates were selected and the susceptibility of MRSA to luteolin and vancomycin was tested by the K-B disc diffusion method. The Minimal Inhibitory Concentrations (MICs) were determined by the microdilution method. Based on MICs,the Fractional InhibitoryConcentration (FIC) was determined bythe chessboard method.The Mutant Prevention Concentrations (MPCs) were measured by agar dilution method and the Selection Index (SI) and frequency of drug resistance was calculated. The effect of luteolin combined with vancomycin on MRSA proliferation was determined by spectrophotometry. Results: There were significant differences in MICs before and after the combination of luteolin and vancomycin (p<0.05). After the combination of luteolin and vancomycin, the FIC was mainly in the range of FIC<0.5 and 0.5<FIC<1.0, mainly showing the synergistic effect (86.7%), followed by the additive effect (13.3%). It was determined that there was no indifferent effect and antagonistic effect. Compared with those of luteolin and vancomycin alone, both the MPC and SI were decreased when luteolin was combined with vancomycin and the frequency of drug resistance also showed a downward trend, displaying significant differences (p<0.05). Compared with 1/4 MIC luteolin and 1/4 MIC vancomycin alone, the combination of 1/4 MIC luteolin and 1/4 MIC vancomycin significantly inhibited the proliferation and growth of MRSA (p<0.05). Conclusion: Luteolin has a certain anti-MRSA activity in vitro. Combined with vancomycin, luteolin can enhance the antibacterial activity of vancomycin, exert a synergistic or additive bacteriostatic effect, reduce the MPC of vancomycin alone and narrow the mutant selection window.

目的 观察不同剂量注射用泮托拉唑钠在中国健康志愿者体内的药代动力学和药效学特征,并评价其安全性.方法 用单中心、随机、开放、平行对照的试验设计,筛选20名健康受试者,采用男女分层区组随机化方法分组,分别静脉滴注40、80 mg注射用泮托拉唑钠(每日2次,连续5d),给药后监测24 h胃内pH值.用LC-MS/MS法测定左旋泮托拉唑钠血药浓度,数据分别用WinNonlin 6.4软件和SAS软件分析处理.结果 首次静脉滴注注射用泮托拉唑钠后,40、80 mg剂量组后左旋泮托拉唑的主要药动学参数:Cmax分别为(2.37±0.61)、(4.56±0.89)tg/mL;AUC(o-t)分别为(4.96±3.26)、(10.16±3.16) μg·h/mL;AUC(o-∞)分别为(5.17±3.68)、(10.34±3.35) μg·h/mL;t1/2分别为(1.67±0.76)、(1.75 ±0.57)h;Tmax分别为(0.50±0.01)、(0.53±0.08)h;连续给药5d后40、80 mg剂量组左旋泮托拉唑的主要药动学参数:Cmax分别为(2.30±0.62)、(4.50±0.71) μg/mL;AUC(0-t)分别为(5.02±3.85)、(10.75±3.82) μg·h/mL;AUC(0-∞)分别为(5.26±4.35)、(10.98±4.10) μg/mL·h;Cmin,ss分别为(0.03 ±0.08)、(0.05 ±0.07)μg/mL;t1/2分别为(1.72±0.78)、(1.80±0.64)h;Tmax分别为(0.52±0.04)、(0.50±0.00)h.用药第1天后40、80 mg剂量组胃内pH ＞4的时间比例分别为(38.6±20.9)％、(46.9±16.5)％;pH ＞6的时间比例分别为(25.5±20.6)％、(32.7±7.9)％;用药第5天后胃内pH ＞4的时间比例分别为(69.6±15.4)％、(80.2±16.6)％;pH ＞6的时间比例分别为(48.3±13.7)％、(49.7±15.7)％.结论 注射用泮托拉唑钠在健康人体内符合线性药代动力学特征.通过两剂量组药效学参数比较,80 mg剂量组的抑酸效果明显优于40 mg剂量组,且服药后较安全.建议临床使用80 mg每12小时1次.

目的:了解尿路感染的病理机制和治疗药物的研究进展,以期为其临床诊断、药物治疗和研发提供参考.方法:查阅近年来国内外相关文献,就尿路感染的类型、病理机制和治疗药物进行归纳和总结.结果与结论:尿路感染主要为尿路致病菌感染,其治疗药物的研究方向主要为以细菌附着、细菌毒素和蛋白酶、铁载体、脲酶、菌毛为靶标的疫苗或小分子药物.以细菌附着为靶标的疫苗可很好地阻断病原体与机体的相互作用,降低尿路感染的发生率;以细菌毒素和蛋白酶为靶标的疫苗的临床疗效仍有待进一步评估;以铁载体为靶标的疫苗可减缓疾病进展和减弱细菌定植;以脲酶为靶标的小分子药物具有一定疗效的同时也存在较严重的副作用;以菌毛为靶标的小分子药物通过阻断尿路感染动物模型的病原体在膀胱上的定植和侵入,能较好并快速地发挥预防和治疗尿路感染的作用.黏附素FimH的抗体已进入了Ⅰ期临床试验阶段,但Pilicides、Mannosides和靶向于菌毛、铁载体以及其他毒力因素的疫苗等仍处于实验研究或临床前研究阶段.

Objectives: Pseudomonas aeruginosa is one of the most common pathogens causing nosocomial pneumonia. The aim of this study was to investigate the epidemiology and resistance of P. aeruginosa isolated from hospitalised patients in the respiratory department of a hospital in China.Methods: Clinical isolates were collected from the respiratory department of Southwest Hospital (Chongqing, China) from January 2013 to December 2014. Patients' social and demographic information was obtained from the hospital's information system. Antimicrobial susceptibility was assessed using the agar dilution method. Screening for carbapenemase production among carbapenem-resistant P. aeruginosa was performed using the modified Hodge test and MBL Etest, and carbapenemase-encoding genes were amplified by PCR. Amplification and sequencing of the oprD gene were performed for carbapenem-resistant P. aeruginosa. Sequence types were determined by multilocus sequence typing (MLST).Results: A total of 92 P. aeruginosa isolates were collected from patients in the respiratory department, and piperacillin/tazobactam was the most effective antibiotic against these isolates. Multivariate analysis revealed that antibiotic use prior to admission was an independent risk factor for P. aeruginosa infection. The isolates comprised 25 genotypes, the most common of which were ST235 and ST111. The bla(IMP-4) gene was present in 4 isolates and bla(VIM-2) in 1 isolate among the 24 carbapenem-resistant isolates. Most of the carbapenem-resistant isolates contained mutational inactivation of the oprD gene.Conclusions: These results suggested that patients and the hospital environment were sources of P. aeruginosa in nosocomial infections. Mutational inactivation of the oprD gene might be the main mechanism of carbapenem resistance. (C) 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.

Pseudomonas aeruginosa (P. aeruginosa) is a common pathogen in hospital-acquired infection and is readily able to form biofilms. Due to its high antibiotic resistance, traditional antibacterial treatments exert a limited effect on P. aeruginosa biofilm infections. It has been indicated that hyperoside inhibits P. aeruginosa PAO1 (PAO1) biofilm formation without affecting growth. Therefore, the current study examined the biofilm formation and quorum sensing (QS) system of PAO1 in the presence of hyperoside. Confocal laser scanning microscopy analysis demonstrated that hyperoside significantly inhibited biofilm formation. It was also observed that hyperoside inhibited twitching motility in addition to adhesion. Data from reverse transcription-quantitative polymerase chain reaction indicated that hyperoside inhibited the expression of lasR, lasI, rhlR and rhlI genes. These results suggest that the QS-inhibiting effect of hyperoside may lead to a reduction in biofilm formation. However, the precise mechanism of hyperoside on P. aeruginosa pathogenicity remains unclear and requires elucidation in additional studies.

OBJECTIVE:To provide reference for the reform of the postgraduate training mode of pharmaceutical professional degree. METHODS:Domestic literatures about postgraduate training mode of pharmaceutical professional degree from 2010 to 2013 were searched and combined with the current education situation in China to conclude and summarize the existing problems and reform strategy. RESULTS:Postgraduate education mode of pharmaceutical professional degree in China was still in the explor-atory stage,with shortage of teachers,unreasonable courses,unsound educational structure,nonsupport of the students,imperfec-tion evaluation systems,and so on. Further promoting the reform of postgraduate education mode needs to strengthen teacher teams ’construction,implement cooperative training;improve training program and teaching courses;strengthen dissertation applica-tion-oriented and practice base construction;fully mobilize the enthusiasm and motivation of students;promote the connection of pharmaceutical professional training and vocational qualifications;and improve the quality assurance systems. CONCLUSIONS:Further implementation of postgraduate training mode of pharmaceutical professional degree and postgraduate education of pharma-ceutical professional degree can train the applied talents that meet social needs,improve employment rate of postgraduates and re-duce waste of educational resources.

To characterize a bla(DIM-2)-carrying 409 kb megaplasmid p12969-DIM of Pseudomonas putida 12969 from a patient with pneumonia in China.The complete nucleotide sequence of p12969-DIM was determined with a paired-end library and a mate-pair library using next-generation sequencing technology.bla(DIM-2), a close bla(DIM-1) variant, was identified in p12969-DIM. DIM-2 differs from DIM-1 by two amino acid substitutions Ser119Leu and Ser209Pro. The p12969-DIM backbone is highly similar to pOZ176, but the IncP-2-type stability/replication/conjugal transfer system in the pOZ176 backbone is absent from p12969-DIM. The p12969-DIM accessory regions, a 45.7 kb MDR and a novel insertion sequence, ISPpu23, are almost entirely distinct from pOZ176. The MDR region contains a novel Tn21-subgroup transposon Tn6286 inserted with two class 1 integrons, In1225 and In1226; a Tn5503-family transposon-like element inserted with a strAB locus; and a novel Tn21-subgroup transposon-like element inserted with a class 1 integron, In1224. The three integrons carry bla(DIM-2) as well as a number of additional genes conferring resistance to quinolones, aminoglycosides, chloramphenicol, florfenicol, trimethoprim, streptomycin, quaternary ammonium compounds and sulphonamides. p12969-DIM has two distinct replication/stability systems, repA/parAB-parB2 of an unknown incompatibility group in the backbone and repABC/mazFE of the IncQ2 group in the MDR region.The MDR region of p12969-DIM harbours many resistance genes as well as a second replication/stability system. This article is the first report of a fully sequenced bla(DIM)-carrying plasmid.

目的 研究亚抑菌浓度环丙沙星对泌尿系统感染常见致病菌大肠埃希菌生物膜形成的影响,为临床抗菌药物的合理应用提供理论依据.方法 采用标准琼脂平板倍比稀释法检测大肠埃希菌最低抑菌浓度,采用PCR扩增检测qnr基因,96孔板结晶紫染色法分析生物膜形成能力.结果 21株大肠埃希菌临床分离株除对阿米卡星和碳青霉烯类药物敏感性较高外,对其它抗菌药物均具有较高的耐药性.15株耐环丙沙星大肠埃希菌qnrB、qnrS和qnrA基因检出率分别为60.0％、40.0％和26.7％.亚抑菌浓度环丙沙星对16株(76.2％)大肠埃希菌生物膜形成具有抑制作用,无诱导作用出现,对大肠埃希菌生物膜形成的影响与耐药性和qnr基因均无相关性.结论 亚抑菌浓度环丙沙星对大肠埃希菌生物膜形成的抑制作用与细菌耐药性和qnr基因均无相关性.

The aim of this study was to analyse the resistance and epidemiological data of 117 Acinetobacter baumannii isolates from Southwest Hospital, Chongqing, China. Except for polymyxin B, tigecycline, minocycline, cefoperazone/sulbactam, amikacin and levofloxacin, the resistance rates of other antimicrobial agents were above 90%. All the clinical isolates had the bla(OXA-51) gene and 114 isolates had the bla(OXA-23) gene. Forty-nine isolates were found to contain the bla(IMP-4) gene. PFGE data showed that 117 isolates were divided into 25 groups. Sixty-three (53.85%) were found to carry the class 1 integron, and the sequence analysis of the class 1 integron internal variable regions - five types, one of which had the bla(IMP-4) gene. For the bla(IMP-4) positive strain without class 1 integron, we found the flanking sequence had the TnpA gene. The result suggested that the resistance gene was widely distributed in our hospital; moreover, the modes of presence and transmission are different and complicated. The results of our study can improve the infection empirical treatment method and infection control programme.

To develop a population-based pharmacokinetic model for the oral antiepileptic drug zonisamide using a cohort of healthy (nonepileptic) subjects and evaluate the effect of individual factors on the pharmacokinetics of zonisamide. 30 young adults (21-39 years) in good health were randomly assigned to 3 equal groups (1:1 sex ratio) for single-dose administration of zonisamide at 200 mg, 300 mg, or 400 mg. An additional 9 subjects (22-24 years) were administered once daily zonisamide at 300 mg for 14 days, and comprised the multiple dosing group. Venous blood samples were collected for analysis prior to (baseline, 0 hours) and after (1-300 hours) drug administration, providing 607 total samples used to build the pharmacokinetic model. The population pharmacokinetic analysis was performed by ICON's nonlinear mixed-effect modeling (NONMEM) software. Validation of the final model was carried out by nonparametric bootstrapping and visual predictive check. The zonisamide pharmacokinetics was best described by a two-compartment model with first-order elimination. In the final model, the estimated value of clearance (CL) was 23.25 L/h, the volume of distribution of the central compartment (Vc) was 34.50 L, the intercompartmental clearance (Q) was 20.22 L/h, and the Ka was 0.026 h(-1). The peripheral volume of distribution (Vp) was 1,429 L for single dose and 1,003 L for multiple doses. Body weight was the significant covariate affecting CL, Vc, Vp, and Q. Otherwise, female subjects had a lower Q than male subjects. The pharmacokinetics of zonisamide after oral administration could be described using a linear first-order elimination two-compartment model, which may provide a reference for clinical use of zonisamide in Chinese adults.

There is a growing body of evidence supporting the emergence of azole resistance in Aspergillus and Candida spp. and this may be of particular concern due to the potential implications in the management of invasive fungal infections occurring in immunocompromised hosts. The aim of present review was to describe the magnitude of the problem, summarising the epidemiology and potential impact of yeast and mould antifungal resistance in patients with haematological malignancies. The first cases of triazole-resistant Aspergillus fumigatus isolates were reported in 1997 in patients receiving itraconazole. More recently, a worrisome increase in the frequency of azole resistance has been reported, primarily in patients with chronic forms of pulmonary aspergillosis. However, estimates of azole resistance in haematological patients are poorly characterised, although widespread use of antifungal prophylaxis might favour the emergence of resistant isolates in this setting. A French study estimated a prevalence of 0.85% azole resistance among 118 A. fumigatus isolates collected in 89 haematological patients. More recently, the epidemiology of azole resistance in A. fumigatus in a cohort of 762 haematological patients who received an allogeneic haematopoietic stem cell transplantation (HSCT) in two German centres has been reported. A. fumigatus was identified in 27 HSCT recipients, and 8 patients (30%) had azole-resistant invasive aspergillosis. In summary, the rate of azole-resistant isolates in patients with haematological malignancies appears to be low; however, the paucity of data currently available requires further prospective surveillance programmes.

BACKGROUND:In recent years, the rising rates of resistance to antimicrobial drugs among pathogens have caused great difficulty for clinicians treating infectious diseases. The aim of the study was to assess the curative effect of fosfomycin in treating urinary tract infections (UTIs) in China.METHODS:We collected clinical isolates of UTIs to determine their susceptibility to fosfomycin by the agar dilution method and to analyze extended-spectrum β-lactamase (ESBL)-producing isolates by the double-disc method on Mueller-Hinton agar. Fosfomycin-modifying enzyme analysis was conducted by PCR. Differences between the different groups were determined by the χ(2) test.RESULTS:We collected 433 UTI isolates, and the result showed that the susceptibility rates of clinical isolates were all above 80%. Only Klebsiella pneumoniae was fosA positive, with a positive rate of 26.7%. No correlation was found for the resistance between the antibiotic drugs tested and fosfomycin in the other bacteria, except for cefepime, levofloxacin and ciprofloxacin in Enterobacter cloacae and imipenem in K. pneumoniae.CONCLUSION:Our data suggest that fosfomycin may be a suitable antimicrobial agent for UTI isolates and ESBL-producing bacteria in our hospital.

Objective:To establish an HPLC method for the determination of zonisamide in human plasma, and study the pharmacokinetics of zonisamide dispersible tablets with single and multiple doses in healthy Chinese volunteers. Methods:Thirty healthy volunteers were randomly divided into three groups. The volunteers in the three groups were given single-dose of zonisamide dispersible tablets respectively with 200, 300 and 400 mg, and then those subjects in 300 mg single-dose zonisamide dispersible tablets group were enrolled for multiple-dose study, who were given zonisamide once a day for fourteen days. The plasma concentrations of zonisamide were determined by HPLC method, the pharmacokinetic parameters were calculated and analyzed by DAS 2.1.1 software. Results:AUC0-t, AUC0-∞and Cmax of zonisamide increased in a linear manner at the dose range from 200 to 400 mg. For the multiple-dose administration, the parameters were as follow:Cmax (27.305 ± 5.201) μg·mL-1, tmax (2.556 ± 0.726) h, t1/2 (59.286 ± 8.882) h, AUC0-t (2 435.713 ± 668.845) μg·h·mL-1, AUC0-∞(2 522.230 ± 720.554) μg·h·mL-1. Conclusion:Cmax showed signiifcant difference in physiological disposition between the male and female after administration of a single-dose. After the administration of multiple-dose zonisamide, steady state plasma concentrations can be achieved.

Objective:To evaluate the pharmacokinetics and bioequivalence of two kinds of voriconazole tablets in healthy Chinese male volunteers. Methods:Nineteen healthy male volunteers were given a single crossover oral dose of voriconazole test preparation and reference preparation (200 mg). The blood samples were collected at 0, 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, 12, 24, 36 hours after administration. The concentrations of voriconazole in plasma were determined by HPLC. The pharmacokinetic parameters and the relative bioequivalence were calculated by DAS Ver 3.0 software. Results:The main pharmacokinetic parameters of test tablet and reference tablet of voriconazole were as follows:AUC0-t (5 304.967±3 072.252), (5 130.968±2 985.462) ng·mL-1·h;AUC0-∞(5 683.143±3 438.261), (5 445.115±3 181.161) ng·mL-1·h;Cmax (925.728±356.11), (1 040.251±448.929) ng·mL-1;tmax (1.566±0.975), (1.382±0.959) h;t1/2 (6.316±2.938), (5.966±2.276) h, respectively. The relative bioavailability of test tablet was (100.5±3.4)%. Conclusion:Two kinds of voriconazole tablets are bioequivalent in healthy Chinese male volunteers.

Objective: To establish an HPLC method for the determination of amisulpride levels in human plasma, determine the amisulpride plasma concentration in healthy volunteers within 48 hours after oral administration of amisulpride tablets,and study on pharmacokinetics. Methods: The amisulpride was extracted from plasma by 10% trichloroacetic acid. Chromatographic separation has been achieved on C18 column with sodium heptanesulfonateacetic acid(pH 4. 85) and methyl cyanides(80∶ 20) as the mobile phase. The fluorescence detection was used to determine amisulpride at λ ex = 280 nm and λ em = 370 nm. Results: The linear range was 5800 ng · mL-1(r = 0. 9997). The average method recovery of low,middle and high concentration was 105. 1%,105. 4%,and 101. 3%,respectively. RSDs of inter-and intra-day precision were both less than 4%. AUC 0-∞ was(3545. 74 ± 1199. 58) ng· h · mL-1,AUC 0-t was(3348. 50 ± 1161. 27) ng · h · mL-1,C max was(457. 69 ± 232. 62) ng · mL-1,T max was(4. 15 ± 1. 31) h,t 1/2 was(10. 77 ± 5. 77) h. Conclusion: The method is simple,rapid,economical and accurate for determination of amisulpride in human plasma and pharmacokinetic study.

The aim of this study was to assess and compare the pharmacokinetic properties, bioavailability, and bioequivalence of a newly developed tablet of voriconazole with those of an established branded formulation in healthy Chinese adult male volunteers. An open-label, single-dose, randomized, 2-way crossover study was conducted in fasted healthy Chinese male volunteers. Eligible participants were randomly assigned in a 1:1 ratio to receive one tablet (200 mg) of the test or reference formulations, followed by a 1-week washout period and administration of the alternate formulations. Plasma samples were collected over 36 hours and analyzed by HPLC. The Voriconazole plasma concentration–time curves were used to obtain pharmacokinetic parameters including AUC0–t, AUC0–∞, Cmax and Tmax. The criteria for bioequivalence were 90% CIs of 80% to 125% for AUC and 70% to 143% for Cmax, and no significant differences for Tmax with a non-parameter test, according to guidelines of the SFDA of China. Tolerability was based on the recording of Adverse Events (AEs). A total of 19 volunteers were included in the study. The mean (SD) Cmax, Tmax, AUC0–t, and AUC0–∞ values after administration of the test and reference formulation, respectively, were as follows: 925.73(356.11) versus 1040.25(448.93) ng/mL, 1.57(0.98) vs 1.38(0.96) hours, 5304.97(3072.25) vs 5141.63(2976.92) ng/mL/h, and 5783.21(3266.86) vs 5520.69(3148.42) ng/mL/h. The relative bioavailability of the test formulation was 103.2% by mean AUC0–t and 104.8% by mean AUC0−∞. The 90% CIs for the ratios of Cmax, AUC0–t, and AUC0−∞ were 77.3% to 122.7%, 85.7% to 114.3%, and 83.6% to 116.4%, respectively, meeting the predetermined criteria for bioequivalence. No drug-related AEs were observed. In this study the test and reference formulations had similar PK parameters and similar plasma concentrationtime profiles and the test formulations met the regulatory criteria for bioequivalence to the established reference formulations based on the rate and extent of absorption. Both formulations were well tolerated.