Peiyuan Xia

Enhancement of Biofilm Formation by Subinhibitory Concentrations of Macrolides in icaADBC-Positive and -Negative Clinical Isolates of Staphylococcus epidermidis

ABSTRACT Biofilm formation in Staphylococcus epidermidis is mediated by icaADBC-dependent and -independent pathways. Subinhibitory concentrations of erythromycin, azithromycin, and clarithromycin enhanced, in a dose-dependent manner, the level of biofilm formation by 20% (21/105 isolates) by macrolide-resistant ica-positive and -negative isolates tested in vitro. The presence of ica, however, apparently produced an enhanced effect on biofilm formation. The levels of expression of the biofilm-related genes icaA, atlE, fruA, pyrR, sarA, and sigB were increased in response to erythromycin. The results likely underscore the potential clinical relevance of macrolide-induced biofilm growth.

Hyperoside attenuates hydrogen peroxide-induced L02 cell damage via MAPK-dependent Keap1–Nrf2–ARE signaling pathway

The flavonoid hyperoside has been reported to elicit cytoprotection against oxidative stress partly by increasing the activity of antioxidant enzymes, such as glutathione peroxidase, superoxide dismutase and catalase. However, the cellular and molecular mechanisms underlying this effect remain unclear. Here, hepatic L02 cells exposed to H(2)O(2) (100 μM) were used to demonstrate that hyperoside protected cells by significantly inhibiting overproduction of intracellular ROS, depletion of the mitochondrial membrane potential and leakage of lactate dehydrogenase. Hyperoside further enhanced the cellular antioxidant defense system through increasing the activity of heme oxygenase-1 (HO-1), and by up-regulating HO-1 expression. Meanwhile, real time PCR, western blot and immunofluorescence studies revealed that hyperoside stimulated nuclear translocation of the Nrf(2) transcription factor in a dose-dependent manner, and this effect was significantly suppressed by pharmacological inhibition of the mitogen-activated protein kinases (MAPK) p38 and ERK. Collectively, our data provide the first description of the mechanism underlying hyperosides ability to attenuate H(2)O(2)-induced cell damage, namely this compound interacts with the MAPK-dependent Keap(1)-Nrf(2)-ARE signaling pathway to up-regulate HO-1 expression and enhance intracellular antioxidant activity.

Spectrum and drug resistance of pathogens from patients with burns

Microbial infection is an obstacle of burn treatment. However, little is known on what types of microbial infection dominate in the burn center and how the dynamic change of those microorganisms occurs during the past several years in China. We conducted a retrospective study of nosocomial infection (NI) in a large burn center to analyze the spectrum and antimicrobial resistance of microbial isolates from January 2003 to December 2010. We studied 989 isolates from 677 patients who had signs and symptoms of infection 48h after admission. The number of NIs per 100 admissions was 10.9. The commonest isolates were Pseudomonas aeruginosa (23.1%), Staphylococcus aureus (18.7%), and Candida (11.4%). The result indicated that the numbers of patients with Acinetobacter sp. infection increased (P=0.004), but with Proteus mirabilis infection decreased (P=0.004). The isolated Acinetobacter sp. and P. aeruginosa were consistently highly resistant to almost all antibiotics tested. Notably, more frequent Acinetobacter sp. isolates appeared to be resistant to amikacin, gentamicin, tobramycin, ceftazidim, piperacillin, tazobactam, levofloxacin, and ciprofloxacin and more frequent Escherichia coli isolates were resistant to ceftazidime and aztreonam at the late time period although the P. aeruginosa and E. coli isolates were sensitive to less used ciprofloxacin and piperacillin/tazobactam. The increased rates of drug-resistant isolates in the later period might be associated with regular prophylactic therapy with antibiotics.

A synthetic cyclic peptide derived from Limulus anti-lipopolysaccharide factor neutralizes endotoxin in vitro and in vivo

Endotoxin, also known as lipopolysaccharide (LPS), is the major mediator of septic shock due to Gram-negative bacterial infections. Recently, much attention has been focused on cationic peptides which possess the potential to detoxify LPS. Limulus anti-LPS factor (LALF), a protein found in the horseshoe crab (Limulus polyphemus), has been proved with striking anti-LPS effects. We synthesized a cyclic peptide (CLP-19), and then investigated its bioactivity both in vitro and in vivo. The ability of CLP-19 to neutralize LPS in vitro was tested using a Limulus amebocyte lysate (LAL) assay and the LPS-binding affinity was measured with an affinity biosensor method. The synthetic peptide LALF31-52 (residues 31 to 52 of LALF) was used as the positive control peptide in this study. It was found that CLP-19 exhibited the significant activity to antagonize LPS without observable cytotoxicity effect on mouse macrophages. CLP-19 directly bound to LPS, and neutralized it in a dose-dependent manner in the LAL assay. Moreover, CLP-19 also showed the remarkable ability to protect mice from lethal LPS attack and to inhibit the LPS-induced tumor necrosis factor alpha (TNF-alpha) release by decreasing serum LPS in vivo. Our work suggests that this peptide is worthy of further investigation as a potential anti-LPS agent in the treatment of septic shock.

Astragalus polysaccharide ameliorates ionizing radiation-induced oxidative stress in mice

Radioprotective compounds from plant resources may represent safe and cost-effective prophylactic and therapeutic agents. This study was designed to investigate the protective effect of polysaccharide derived from the dried roots of the Astragalus spp. (APS) against ionizing radiation (IR) injury in liver and to explore its role in radiation-induced oxidative stress using a mouse model. Prior to (60)Co γ-irradiation (5Gy, single dose), mice received 7 days of APS at low, mid and high doses (50, 100 or 200mg/kg/day, respectively; n=6 each group), vehicle alone (5mL normal saline orally/daily; n=6). A non-irradiated control group (n=6) received the 7-day distilled water regimen only. At 24h post-irradiation, the APS pre-treated mice showed significantly decreased alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase levels, and NF-κB expression. All APS-treated mice also showed attenuation of the IR-induced increase in thiobarbituric acid reactive substance and resolution of the IR-induced decreases in superoxide dismutase, catalase and glutathione activities (all p<0.05). High dose APS pre-treatment led to remarkably less morphologic features of IR-induced hepatic and pulmonary injury. Thus, APS exerts protective effects against IR-induced injury in liver in mice, and the related molecular mechanism may involve suppressing the radiation-induced oxidative stress reaction.

Astragalus polysaccharides protect against dextran sulfate sodium-induced colitis by inhibiting NF-κВ activation

Astragalus polysaccharide (APS) is a bioactive extract of Astragalus membranaceus (AM), which possess a wide range of medicinal benefits, including anti-inflammatory, anti-oxidative, anti-tumor and anti-diabetic effects. The present work evaluated the therapeutic effect of APS and its potential mechanisms in a mouse model of dextran sulfate sodium (DSS)-induced colitis. The APS treatment led to significant improvements in colitis disease activity index (DAI) and histological scores, as well as significantly increased weight and colon length in mice as compared to the control group. Mechanically, reduced NF-κВ DNA phosphorylation activity and downregulated TNF-α, IL-1β, IL-6, IL-17 expressions and myeloperoxidase (MPO) activity were associated with improvement in colitis observed in APS-treated mice. These findings suggest that APS may represent a natural therapeutic approach for treating inflammatory bowel disease, such as ulcerative colitis.

Epidemiology of CTX-M-type extended-spectrum beta-lactamase (ESBL)-producing nosocomial - Escherichia coli infection in China.

BackgroundEscherichia coli is one of the most common clinical pathogens causing nosocomial infection. The widespread cefotaxime-beta lactamases (CTX) has increased the multidrug resistance (MDR) of E. coli and has brought great trouble to the doctor treating the infection.MethodsESBL-positive E. coli isolates were collected from different hospitals in different areas and the minimal inhibitory concentration (MIC) was analyzed by the agar dilution method. The resistance gene types were detected using polymerase chain reaction (PCR) and the sequence types were determined by multilocus sequence typing (MLST).ResultsWe found that the blaCTX-M-1 group and the blaCTX-M-9 group were the main CTX-M gene types, with many kinds of MLST gene types. Except for TEM with high isolate, SHV, OXA and VEB were relatively rare, while no PER and GES was detected. Most strains may have other resistance mechanisms, and the ESBL positive strains have high resistance not only to cephalosporins but also to other kinds of antibiotics.ConclusionThe study provides wide epidemiological data and enables more effective infection control and treatment plans.

Lipopolysaccharide (LPS) detoxification of analogue peptides derived from limulus anti-LPS factor.

Lipopolysaccharide (LPS) plays a critical role in the pathogenesis of sepsis due to gram-negative bacterial infections. Therefore, LPS-neutralizing molecules could have important clinical applications. Our previous work showed, CLP19, an analogue peptide derived from limulus anti-LPS factor (LALF), possessed the capacity to neutralize LPS and thereby inhibit the LPS-induced responses. However, potential cytotoxicity of CLP19 was also found, especially when added to human red blood cells. Accordingly we further developed two peptides (designated as CLP19-1 and CLP19-2) by single- and double-point amino acid substitution of CLP19, respectively, in order to reduce its toxicity and meanwhile retain the anti-LPS activity. In this study, the LPS-detoxifying effectiveness of these peptides was evaluated both in vitro and in vivo. CLP19-1 was found to dose-dependently neutralize LPS in vitro, with significantly lower hemolysis of red blood cells as compared with CLP19. Further in vivo tests verified that CLP19-1 exerted significant protective effects on mice against LPS, characterized by significantly improved survival, decreasing of tumor necrosis factor alpha (TNF-α) serum level and alleviation of tissue injury. Our work indicates that CLP19-1 is worthy of further study as potential anti-LPS agents for the management of sepsis.

Development and validation of liquid chromatography-tandem mass spectrometry method for simultaneous determination of six steroidal saponins in rat plasma and its application to a pharmacokinetics study.

A specific and reliable liquid chromatography-electrospray ionization-tandem mass spectrometry method was developed for the simultaneous determination of timosaponin H1 (TH1), timosaponin E1 (TE1), timosaponin E (TE), timosaponin B-II (TB-II), timosaponin B-III (TB-III) and anemarrhenasaponin I (AS-I) in rat plasma. After addition of internal standard (IS) ginsenoside Rh1, plasma samples were pretreated by protein precipitation with acetonitrile. Chromatographic separation was performed on a reverse phase ACQUITY™ BEH C18 column (100mm×2.1mm i.d., 1.7μm) using a gradient mobile phase system of acetonitrile-water containing 0.05% formic acid and 5mM ammonium formate. The triple quadruple mass spectrometer was set in negative electrospray ionization mode and multiple reaction monitoring (MRM) was used for six steroidal saponins quantification. The precursors to produce ion transitions monitored for TH1, TE1, TE, TB-II, TB-III, AS-I and IS were m/z 1211.5>1079.6, 935.5>773.4, 935.4>773.5, 919.6>757.4, 901.5>739.3, 757.4>595.3 and 637.3>475.3, respectively. The method validation was conducted over the curve range of 0.5-400ng/mL for the six saponins. The intra- and inter-day precisions (RSD%) were less than 9.4% and the average extraction recoveries ranged from 82.5% to 97.8% for each analyte. Six steroidal saponins were proved to be stable during sample storage, preparation and analytical procedures. The validated method was successfully applied for the first time to determine the concentrations of six main steroidal saponins in incurred rat plasma samples, after intragastric administration of the extract of Anemarrhena asphodeloides Bge. for a rat pharmacokinetic study.

Characterization of the Antibacterial Activity and the Chemical Components of the Volatile Oil of the Leaves of Rubus parvifolius L.

Rubus parvifolius L. (Rp) is a medicinal herb that possesses antibacterial activity. In this study, we extracted the volatile oil from the leaves of Rp to assess its antibacterial activity and analyze its chemical composition. A uniform distribution design was used to optimize the extraction procedure, which yielded 0.36% (w/w) of light yellowish oil from the water extract of Rp leaves. We found that the extracted oil effectively inhibited the growth of a wide range of Gram positive and negative bacteria, including Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumanii, Bacillus cloacae, and Klebsiella pneumoniae. We further analyzed the components contained in the hydro-distillated Rp volatile oil by gas chromatography-mass spectroscopy. Twenty nine compounds were identified, including 4-hydroxy-3-methoxystyrene (66%), 3,7,11,15-tetramethyl-2-hexadecen-1-ol (10%) and 4-tert-butylbenzoic acid (2%). Our results suggest that one or multiple constituents contained in Rp volatile oil may account for its antibacterial activity.