Objective To investigate the association of AcrAB-tolC efflux pump and marA-soxS-rob regulon with ciprofloxacin-resistance in Shigella flexneri isolates.Methods Forty five strains of Shigella flexneri were isolated in stool samples collected from outpatient diarrhea clinics in Tianjin from 2009 to 2013.The antimicrobial susceptibility of bacteria was analyzed by Kirby-Bauer disk diffusion method.Ten ciprofloxacin-resistant Shigella flexneri isolates and 10 ciprofloxacin-sensitive isolates were randomly selected.The gyrA and parC genes,plasmid mediated quinolone resistant (PMQR) determinants,efflux pump genes (acrA,acrB) and regulation genes (marA,soxS,rob) were screened and sequenced.Minimum inhibition concentrations (MICs) of the strains were determined before and after efflux pump inhibitor carbonyl cyanide m-chlorophenylhydrazone (CCCP) was added.The expression of acrA,acrB,marA,soxS and rob genes in ciprofloxacin-resistant and ciprofloxacin-sensitive strains was detected by realtime fluorescence quantitative RCR (RT-PCR),and the differences of expression were evaluated using t test.Results Both gyrA and parC mutations were detected in all ciprofloxacin-resistant strains; qnrS1 was positive in ciprofloxacin-resistant strain CR2 and aac(6')-Ib-cr was positive in ciprofloxacin-resistant strain CR5.Efflux pump genes and regulation genes were not detected in ciprofloxacin-sensitive strains,while soxRS mutation was detected in all ciprofloxacin-resistant strains except CR10.MICs of quinolones in ciprofloxacin-resistant strains decreased to one-fourth or one-eighth when CCCP added,while not changed or only decreased to one-half in ciprofloxacin-sensitive strains.Expressions of acrA,acrB,marA and soxS were significantly higher in ciprofloxacin-resistant strains than those in ciprofloxacin-sensitive strains (P ＜ 0.05 or P ＜ 0.01).Conclusion Efflux pump may involve in the high-level ciprofloxacin resistance in Shigella flexneri through activating transcription of efflux pump genes by marA-soxS-rob regulon,in which soxRS mutation may play an important role.

OBJECTIVETo investigate the protective effect and mechanism of sequoyitol (Sep) on high glucose-induced human umbilical vein endothelial cells (HUVECs) injury.

METHODSHUVECs were cultured with high glucose (30 mmol/L) in the presence or absence of sequoyitol (0.1, 1 and 10 micromol/L) for 24 h. Cell proliferation was measured by BrdU marking and cell cycle was detected by flow cytometry. 2', 7'-dichlorofluorescein diacetate was used to evaluate intracellular reactive oxygen species (ROS) levels. The NO, malonydialdehyde (MDA) and H2O2 levels were determined by colorimetric method according to the manufacturer's instructions. The expression of endothelial nitric oxide synthase (eNOS) and NADPH oxidase 4 (NOX4) were measured by real-time PCR and Western blot.

RESULTSIn the present study, we found that sequoyitol pretreatment for 1 h significantly decreased cell injury, promoted cell proliferation. Meanwhile sequoyitol significantly down-regulated NOX4 expression and decreased the level of ROS, MDA and H2O2 and obviously increased NO levels and up-regulated eNOS expression.

CONCLUSIONSequoyitol alleviates high glucose-induced cell injuries in HUVECs via inhibiting oxidative stress and up-regulating eNOS expression.

Cell Proliferation ; Cells, Cultured ; Glucose ; toxicity ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Hydrogen Peroxide ; metabolism ; Inositol ; analogs & derivatives ; pharmacology ; Malondialdehyde ; metabolism ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Oxidative Stress ; Reactive Oxygen Species ; metabolism

The aim of the present study is to investigate the effects of sequoyitol (Seq) on expression of eNOS and NOX4 in aortas of type 2 diabetic rats. Type 2 diabetic rats induced by high fat and high sugar diet and low dose of streptozotocin (STZ, 35 mg x kg(-1)) and were administered Seq (12.5, 25 and 50 mg x kg(-1) x d(-1)) for 6 weeks. The fasting blood glucose (FBG) and body weight were tested. Acetylcholine (Ach) induced endothelium-dependent relaxation and sodium nitroprusside (SNP) induced endothelium-independent relaxation were measured in aortas for estimating endothelial function. Aortic morphological change was observed with HE staining. The level of serum insulin was measured by radioimmunoassay. The total antioxidative capacity (T-AOC), malondialdehyde (MDA) and NO levels in aortas were determined according to the manufacturer's instructions. In addition, the expressions of eNOS and NOX4 in aortas were measured by immunohistochemisty, real-time PCR or Western blotting. The results showed that Seq significantly decreased FBG and insulin resistance, and improved aortic endothelium-dependent vasorelaxation function. The expressions of NOX4 and MDA content were obviously decreased, while the expression of eNOS, the levels of NO and T-AOC increased significantly in aortas of diabetic rats with Seq treatment. In conclusion, Seq protects against aortic endothelial dysfunction of type 2 diabetic rats through down-regulating expression of NOX4 and up-regulating eNOS expression.
Animals ; Aorta ; metabolism ; pathology ; Blood Glucose ; metabolism ; Body Weight ; Diabetes Mellitus, Experimental ; chemically induced ; metabolism ; physiopathology ; Diabetes Mellitus, Type 2 ; chemically induced ; metabolism ; physiopathology ; Hypoglycemic Agents ; pharmacology ; Inositol ; analogs & derivatives ; pharmacology ; Insulin ; blood ; Insulin Resistance ; Male ; Malondialdehyde ; metabolism ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Oxidation-Reduction ; drug effects ; Rats ; Rats, Sprague-Dawley ; Streptozocin ; Vasodilation ; drug effects

Adult ; Bone Neoplasms ; pathology ; surgery ; Femur ; pathology ; surgery ; Humans ; Male ; Osteochondroma ; pathology ; surgery ; Young Adult

OBJECTIVETo introduce the framework of evidence-based practice with a case of castration-resistant prostate cancer (CRPC) as an example.

METHODSA clinical question was formulated according the clinical scenario. A systematic search was conducted for the published literature in the databases of PubMed, EMBASE, Cochrane Library, Clinical Trial Registries, and Web of Knowledge up to Dec 2014. The identified literature was reviewed for quality appraisal before the evidence was applied to clinical practice.

RESULTSThe treatment was effective and the patient achieved disease remission.

CONCLUSIONEvidence-based practice should be integrated with clinical scenario, current evidence, and patients' willingness, and follow a systematic framework.

Evidence-Based Medicine ; Humans ; Male ; Orchiectomy ; Prostatic Neoplasms, Castration-Resistant ; therapy

To establish an HPLC-MS/MS method for the analysis of quercetin, kaempferid and isorhamnetin in rats plasma and study its pharmamacokinetics after an intragastrical administration of Hippophae rhamnoides extracts. Five healthy male Sprague-Dawley (SD) rats were given single doses of H. rhamnoides extracts (quercetin 26.35 mg x kg(-1), kaempferid 4.040 mg x kg(-1), isorhamnetin 31.37 mg x kg(-1)), and then their orbital sinus blood samples were collected at different time points. The drug plasma concentration of the three flavonoids was determined by HPLC-MS/MS method. After that, the main pharmacokinetics parameters were calculated by using Kinetica 5. 0. 11 software. The methodological test showed that the linear concentration ranges of quercetin, kaempferid and isorhamnetin were 7.500-600.0 μg x L(-1) (R2 = 0.998 5), 1.000-80.00 μg x L(-1) (R2 = 0.998 5 ) and 10.00-800.0 μg x L(-1) (R2 = 0.998 0), respectively. The inner and inter-days precisions were both less than 14.0%. The plasma samples showed a good stability and consistency with the requirement of biological sample analysis after the samples were frozen once and placed at - 20 degrees C for 15 d and room temperature for 6 h and the treated analytes were placed at -20 degrees C for 24 h. For quercetin, the pharmacokinetic parameter t(½β), AUC(0-∞), MRT(0.∞), C.(max) and T(max) were (113.3 ± 19.37) min, (12 542.14 ± 3 504.05) μg x h x L(-1), (119.6 ± 13.29) h, (164.6 ± 27.33) μg x L(-1) and (5.199 ± 0.840 3) h, respectively. For kaempferid, the pharmacokinetic parameters t(½β), AUC(0-t), MRT(0-∞), C(max) and T(max) were (79.85 ± 17.15) min, (934.51 ± 94.59) μg x h x L(-1), (81.50 ± 13.75) h, (80.15 ± 14.24) μg x L(-1) and (3.827 ± 0.902 7) h, respectively. For isorhamnetin, the pharmacokinetic parameters t1,2,, AUC(0-t), MRT(0-∞), C(max) and T(max) were (118.3 ± 20.73) min, (26 067.77 ± 4 124.60) μg x h x L(-1), (129.0 ± 16.30) h, (269.6 ± 29.32) μg x L(-1) and (6.513 ± 1.450) h, respectively. The HPLC-MS/MS analysis method established in this study was proved to be sensitive and accurate and could be applied in the pharmacokinetic study of quercetin, kaempferid and isorhamnetin in rat plasma.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Hippophae ; chemistry ; Kaempferols ; blood ; pharmacokinetics ; Male ; Quercetin ; analogs & derivatives ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Objective To investigate changes in the healthcare and support needs during the diagnostic period,and factors that affect these needs in women with suspected breast cancer.Methods This study used an investigator-developed,self-administered questionnaire to collect data from 283 women on three occasions:notification cation of need for breast biopsy,before biopsy and after diagnosis.Results The total score of need before the patients told breast biopsy was (27.68 ±0.53 ),and was higher than that after diagnosis ( 26.80 ±0.47) and the highest score was that before biopsy,which was ( 27.93 ± 0.49),and the difference among the three groups was significant ( F=6.48,P ＜ 0.01 ) ; needs score before diagnosis was ( 28.83 ± 0.31 ) and (27.06 ± 0.46) after diagnosis in participants whose education background was senior middle school or above;needs score before diagnosis was (27.04 ± 0.34) and ( 26.92 ± 0.48) after diagnosis in participants whose education background was junior high school or below; the differences was significant ( t=- 44.09,- 2.40,respectively; P ＜ 0.05 ).Conclusions Need levels of women with suspected breast cancer vary during the diagnostic period,and are highest before breast biopsy,and related to personal characteristics and cultural context.Therefore,during this period,nursing staff should provide patients and families with cuhurally sensitive,individualized,supportive care.

No abstract available.
Asian Continental Ancestry Group* ; Ectodermal Dysplasia 1, Anhidrotic* ; Humans

Objective To investigate the role of AcrAB-TolC efflux pump in fluoroquinolones resistance by Shigella. spp and to explore the significance of AcrAB-TolC efflux pump on mutation of acrR, soxS and marOR as well as on drug re?sistence. Methods Drug resistant bacteria were selected by Kirby-Bauer disk diffusion test. After addition of efflux pump inhibitor carbonylcyanide-m-chlorophenylhydrazone (CCCP), change of minimal inhibitory concentration (MIC)s of nilidixic acid, Levofloxacin, ofloxacin, ciprofloxacin and Norfloxacin were examined. The DNA binding region of acrA, acrB, soxS, acrR and marOR gene in these mutants were amplified by PCR and sequenced. Results Among the 159 clinical isolates of Shigella,11 strains are resistant to fluoroquinolone. After the addition of CCCP, MICs of 2 fluoroquinolone resistant strains decreased; the MICs of 7 fluoroquinolone resistant strains did not change; MICs of 2 fluoroquinolone resistant strains in?creased. The corresponding nucleotides C, A, T, T on the 36th to 39th of marOR gene were missing, showing by sequencing, in fluoroquinolone resistent strains which might be regulated by the efflux pump gene AcrAB-TolC. Conclusion Efflux pump inhibitor could restrain the activity of efflux partially. The mutations of marOR might play an important role in fluoroquino?lone resistent by shigella.

Objective To explore carrying rates of class 1, class 2 integrons as well as ISCR1 in Shigella isolates and their connection with drug resistance. Methods Antibiotic sensitivities were detected by K-B disk diffusion in 159 clinical isolates. Total bacteria DNA was prepared through boiling the isolates and the DNA was then used as template for PCR am?plification. PCR, ZSCR1 and sequencing analyse of integrons were applied to all of them. Results were compared by Blast and GenBank. Results Antibiotic sensitivity results showed that in the S. flexneri strains the incidence of resistance to tet?racycline and streptomycin were 88.68%and 81.13%in the S. flexneri strains while the incidence of resistant to chloram?phenicol and trimethoprim-sulfamethoxazol were both 56.60%, and the incidence of multidrug drug resistance was 77.36%. In the sonnei strains, the incidence of resistance to ampicillin, trimethoprim-sulfamethoxazo were 97.17% and 95.28%, 83.96%and 76.42%respectively, and the incidence of multidrug resistance was 98.11%. Among all isolates, 118 were class 1 integron positive , 70 were class 2 integron positive and 89 were double positives. For those 118 isolates that are positive of class 1 integron, 23 were typical while 95 weres atypical. The gene cassettes of typical class 1 integrons contains aadA2, aa?dA1, dfrⅠ, blaoxa-10 and blaoxa-1. IntI1, aadA, blaoxa-1 and IS1 were included in the gene cassetes of the atypical class 1 integrons. Class 2 integrons positive isolates carried gene cassttes which include dfrA1, satl and aadA1. No ISCR1 was found in any isolate. Integron carriage strains were closely associated with higher rate of multiple antibiobic resistance com?pared with the organisms without integrons (90.65%,50%, P<0.05). Conlusion Class 1 and class 2 integrons were widely existence in Shigella isolates and were related to the multidrug resistance.