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Wei Zeng

Publications and source records attributed to Wei Zeng.

At least 37 records · Page 2Linked to original sources
PubMed2006

[Quantification for biofilm formation of pseudomonas aeruginosa isolates and their genotypic analysis].

OBJECTIVE: To probe the correlation between the ability of biofilm formation of Pseudomonas aeruginosa isolates and their genotypes. METHODS: Forty-eight Pseudomonas aeruginosa isolates were tested for their biofilm formation with a modified microtiter test and were analyzed by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR). The percent similarity between their genetic fingerprints and cluster analysis was performed and worked out using computer software. RESULTS: Forty-eight Pseudomonas aeruginosa isolates demonstrated different abilities of biofilm formation in vitro. And the 48 isolates with different abilities of biofilm formation showed different genotypes in fingerprints by ERIC-PCR. Among the 48 isolates of Pseudomonas aeruginosa tested, five clusters (A, B, C, D, E), were identified at the 17% genetic similarity level. The isolates with strong ability of biofilm formation were in D cluster at the 42% genetic similarity level. CONCLUSION: Most of the 48 PA isolates tested formed strong biofilm, and their genotypes were of correlation.

Biofilms↗
PubMed2006

[Establishing a real-time PCR assay for anatid herpesvirus 1].

OBJECTIVES: To develop a real-time quantitative PCR assay to detect duck plague virus (DPV) for the rapid diagnosis of DPV infection, the investigation of its nosogenesis and the screening of effective antiviral drugs. METHODS: The primer and probe were designed according to the gene sequence of DPV DNA polymerase gene. To establish a standard curve, a plasmid containing 125 bp PCR product was constructed and severed as a positive control. The TaqMan based real-time PCR method was adopted and compared with the traditional PCR approach in sensitivity, reliability and specificity. RESULTS: A good linear correlation was demonstrated in the standard curve for the real-time PCR assay within the range from 2.3 x 10 to 2.3 x 10(5) copies. A minimum of 23 positive plasmids could be detected, indicating a good sensitivity of the assay. The coefficients of variance (CVs) were 1.22-6.69 and 2.09-8.84 for the intra-assay and inter-assay tests respectively, indicating a good reliability. No amplification products were found for DNA from other pathogens, indicating a good specificity. The comparative study proved that the TaqMan technology was much more sensitive than traditional PCR assay. CONCLUSION: The real-time quantitative PCR assay for DPV DNA has good sensitivity, specificity and reliability.

Alphaherpesvirinae↗
PubMed2005

Bifunctional AgOAc-catalyzed asymmetric [3 + 2] cycloaddition of azomethine ylides.

[reaction: see text] A bifunctional AgOAc-catalyzed asymmetric cycloaddition of azomethine ylides with electronic-deficient alkenes was developed using ferrocenyloxazoline-derived N,P ligands. The reactive metal-bound azomethine ylide dipole is formed by the deprotonation with acetate, and extra base is not necessary. The reactions proceed with high enantioselectivity. This method provides an efficient and convenient route to optically active pyrrolidine derivatives.

Journal Article↗
PubMed2005

The action of Pseudomonas aeruginosa biofilms in intrinsic drug resistance.

BACKGROUND: There is a growing interest in studying the relationship between intrinsic resistance and biofilms resistance to drugs. However, the relationship still remains unclear in the macroscopic bacterial growth. Our study is to illuminate the change of bacterial drug resistance of gyrA mutant and active efflux pump during the development of Pseudomonas aeruginosa (P. aeruginosa) biofilms. METHODS: The strains of type II topoisomerase gene mutant (gyrA mutant) and multidrug resistance (MDR) efflux pump were clinical isolates and detected by polymerase chain reaction (PCR). The process of bacterial biofilms development was observed by scanning electron microscope. Triparental mating experiments were performed to transfer report gene of green fluorescent protein (GFP) into P. aeruginosa biofilms strains and followed by analysis of bacterial survival rate between intrinsic resistance and biofilms resistance. RESULTS: The fluorescent strains with pGFPuv could develop mature biofilms on Teflon surface. Before a period of 72 hours, the survival rate of biofilms bacteria and intrinsic resistance strains in ciprofloxacin solution was significantly different (P < 0.05). The survival number of intrinsic resistance strains (gyrA mutation and active efflux pump) was illustriously higher than biofilm strain in the initial stage of biofilms development. After 72 hours incubation, there was no clearly difference between mutants and biofilms strains in the survival rate (P > 0.05). The carbonyl cyanide m-chlorophenylhydrazone and azithromycin could significantly reduce the drug resistance of biofilm strains and efflux pump strains. CONCLUSIONS: In the development of P. aeruginosa biofilms, the strains of gyrA mutation and MDR efflux could be conferred with new level of drug resistance. When co-cultured mutated strains with biofilm strains, biofilms may play a major role in bacterial resistance. But after 72 hours incubation (a mature biofilms had been developed), there was no clearly difference between the number of mutant strains and biofilm strains.

Biofilms↗
PubMed2005

Switch movements and the myosin crossbridge stroke.

The myosin II motor from Dictyostelium discoideum has been engineered to contain single tryptophan residues at strategic locations to probe movements of switch 1 and switch 2. The tryptophan residue at W501 probes movement of the relay helix and indirectly reports on switch 2 movement. This probe suggests that there is an equilibrium between the switch 2 open- and closed-states when the gamma-phosphate position is occupied. Actin does not appear to greatly affect this equilibrium directly, but has indirect influence via switch 1. The latter region has been probed by introducing tryptophan residues at positions 239 and 242. The kinetics of the actomyosin ATPase in solution is discussed with respect to recent crossbridge models based on high-resolution crystal structures.

Actins↗
PubMed2005

Protonation, photobleaching, and photoactivation of yellow fluorescent protein (YFP 10C): a unifying mechanism.

Yellow fluorescent protein (YFP 10C) is widely used as a probe in biology, but its complex photochemistry gives rise to unusual behavior that requires fuller definition. Here we characterize the kinetics of protonation and reversible bleaching over time scales of picoseconds to hours. Stopped-flow and pressure-jump techniques showed that protonation of the fluorescent YFP(-) anion state is two-step with a slow transition that accounts for blinking of 527 nm emission at the single molecule level on the seconds time scale. Femtosecond spectroscopy revealed that the protonated excited-state (YFPH*) decayed predominantly by a radiationless mechanism, but emission at 460 nm was detected within the first picosecond. Limited excited-state proton transfer leads to 527 nm emission characteristic of the YFP(-*) anion. Prolonged continuous wave illumination at the peak of YFP(-) absorbance (514 nm) yields, irreversibly, a weakly fluorescent product that absorbs at 390 nm. This "photobleaching" process also gives a different species (YFPHrb) that absorbs at 350/430 nm and spontaneously regenerates YFP(-) in the dark on the time scale of hours but can be photoactivated by UV light to regenerate YFP(-) within seconds, via a ground-state protonated intermediate. Using a pulsed laser for photobleaching resulted in decarboxylation of YFP as indicated by the mass spectrum. These observations are accounted for in a unifying kinetic scheme.

Amino Acid Sequence↗
PubMed2005

Pharmacokinetics and pharmacodynamics of sitagliptin, an inhibitor of dipeptidyl peptidase IV, in healthy subjects: results from two randomized, double-blind, placebo-controlled studies with single oral doses.

BACKGROUND: Sitagliptin (MK-0431 [(2R)-4-oxo-4-(3-[trifluoromethyl]-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7[8H]-yl)-1-(2,4,5-trifluorophenyl)butan-2-amine]) is an orally active, potent, and selective inhibitor of dipeptidyl peptidase IV (DPP-IV) currently in phase III development for the treatment of type 2 diabetes. METHODS: Two double-blind, randomized, placebo-controlled, alternating-panel studies evaluated the safety, tolerability, pharmacokinetics, and pharmacodynamics of single oral doses of sitagliptin (1.5-600 mg) in healthy male volunteers. RESULTS: Sitagliptin was well absorbed (approximately 80% excreted unchanged in the urine) with an apparent terminal half-life ranging from 8 to 14 hours. Renal clearance of sitagliptin averaged 388 mL/min and was largely uninfluenced by the dose administered. The area under the plasma concentration-time curve for sitagliptin increased in an approximately dose-dependent manner and was not meaningfully influenced by food. Single doses of sitagliptin markedly and dose-dependently inhibited plasma DPP-IV activity, with approximately 80% or greater inhibition of DPP-IV activity occurring at 50 mg or greater over a 12-hour period and at 100 mg or greater over a 24-hour period. Compared with placebo, sitagliptin produced an approximately 2-fold increase in postmeal active glucagon-like peptide 1 levels. Sitagliptin was well tolerated and was not associated with hypoglycemia. CONCLUSIONS: This study provides proof of pharmacologic characteristics for sitagliptin in humans. By inhibiting plasma DPP-IV activity, sitagliptin increases the postprandial rise in active glucagon-like peptide 1 concentrations without causing hypoglycemia in normoglycemic healthy male volunteers. Sitagliptin possesses pharmacokinetic and pharmacodynamic characteristics that support a once-daily dosing regimen.

Administration, Oral↗
PubMed2005

[Isolation of Malassezia furfur from the groin abscess of a renal transplant patient].

We report here the discovery of Malassezia furfur from a groin abscess of a renal transplant patient. A 33-year-old male patient was admitted to our hospital because of a high fever and a persistent inflammatory nodule on his right groin for one week. He had received a renal transplant 3 years before and remained on immunosuppressive agents. He was treated with broad-spectrum antibiotics after hospitalization but the nodule formed a large abscess and then a deep ulcer instead of resolving. Examination of the culture by light microscopy revealed ovoid budding yeasts displaying collar-shaped structure. Subculture of the primary colonies onto Sabouraud's dextrose agar and medium containing rapeseed oil resulted in growth only on the medium containing rapeseed oil. All of the isolates was identified as Malassezia furfur. The pathogenicity of the isolates was tested in mice by intravenous injection of (3-5) x 10(8) cfu per mouse after immunosuppression with 500 mg/kg of prednisone intraporitoneally on day-2. In the mouse model, micro-abscess and inflammatory reaction and oval yeasts with budding were noted in histopathologic section of the viscera of the mice. A rib-like or serrate-like structure of the inner side of cell wall, characteristic for Malassezia spp., was observed by transmission electron microscopy. The patient received oral fluconazole and topical amphotericin B. The isolate before antifungal therapy was sensitive to both fluconazole and amphotericin B, while the isolate after antifungal treatment was only sensitive to amphotericin B. Proteinase activity of the isolates increased 1.43 times after antifungal treatment. This case indicated the invasive power of M. furfur in deep infection. Renal transplantation and reception of long-term immunosuppressive treatment are risk factors for the invasive infection of this fungus.

Abscess↗
PubMed2004

Dynamics of actomyosin interactions in relation to the cross-bridge cycle.

Transient kinetic measurements of the actomyosin ATPase provided the basis of the Lymn-Taylor model for the cross-bridge cycle, which underpins current models of contraction. Following the determination of the structure of the myosin motor domain, it has been possible to introduce probes at defined sites and resolve the steps in more detail. Probes have been introduced in the Dicytostelium myosin II motor domain via three routes: (i) single tryptophan residues at strategic locations throughout the motor domain; (ii) green fluorescent protein fusions at the N and C termini; and (iii) labelled cysteine residues engineered across the actin-binding cleft. These studies are interpreted with reference to motor domain crystal structures and suggest that the tryptophan (W501) in the relay loop senses the lever arm position, which is controlled by the switch 2 open-to-closed transition at the active site. Actin has little effect on this process per se. A mechanism of product release is proposed in which actin has an indirect effect on the switch 2 and lever arm position to achieve mechanochemical coupling. Switch 1 closing appears to be a key step in the nucleotide-induced actin dissociation, while its opening is required for the subsequent activation of product release. This process has been probed with F239W and F242W substitutions in the switch 1 loop. The E706K mutation in skeletal myosin IIa is associated with a human myopathy. To simulate this disease we investigated the homologous mutation, E683K, in the Dictyostelium myosin motor domain.

Actins↗
PubMed2004

High-throughput liquid chromatography for drug analysis in biological fluids: investigation of extraction column life.

A novel method was developed and assessed to extend the lifetime of extraction columns of high-throughput liquid chromatography (HTLC) for bioanalysis of human plasma samples. In this method, a 15% acetic acid solution and 90% THF were respectively used as mobile phases to clean up the proteins in human plasma samples and residual lipids from the extraction and analytical columns. The 15% acetic acid solution weakens the interactions between proteins and the stationary phase of the extraction column and increases the protein solubility in the mobile phase. The 90% THF mobile phase prevents the accumulation of lipids and thus reduces the potential damage on the columns. Using this novel method, the extraction column lifetime has been extended to about 2000 direct plasma injections, and this is the first time that high concentration acetic acid and THF are used in HTLC for on-line cleanup and extraction column lifetime extension.

Acetic Acid↗
PubMed2004

Clinical investigation on the correlation between lower urinary tract infection and cystitis glandularis.

In order to study the association between lower urinary tract infection and cystitis glandularis (CG), 120 cases of CG were diagnosed by cystoscopic biopsy in the suspicious foci of the bladder. Among them, 72 cases were subjected to bacterial counting culture of urine and microscopic examination of urinary sediment, and 60 cases to fluorescence quantitative polymerase chain reaction (FQ-PCR) assay to detect HPV, CMV and HSV DNA in urine samples. In the 72 cases of CG, the positive rate of bacterial counting culture of urine was 15.3% (11/72), and gray zone rate was 18.1% (13/72). 31.9% (23/72) patients were positive in bacterioscopy of urinary sediment. There was statistically significant difference as compared with the control group (P<0.01). Only 4 of 60 urine samples were positive by FQ-PCR in detection of the three viruses mentioned above with the positive rate being 6.67%. Compared with the control group, there was no significant difference (P>0.05). It was concluded that the genesis of CG was closely correlated with the chronic lower urinary tract infection, especially caused by Esch coli.

Adult↗
PubMed2004

A two-stage SBR process for removal of organic substrate and nitrogen via nitrite-type nitrification-denitrification.

A two-stage SBR process (TSSBR) was applied for the treatment of wastewater with high strength COD and nitrogen. Most of organic substrate was removed in the first stage SBR reactor (SBR1) under the aerobic condition. Subsequently the second stage SBR reactor (SBR2) firstly was operated under the aerobic condition for simultaneous nitrite-type nitrification and removal of a small amount of residual organic substrate. Nitrification was controlled to the nitrite-type nitrification. Then denitrification in which the wastewater was used as external carbon sources occurred in SBR2 under the anoxic condition. The experimental results indicated that two kinds of biomass with the different function were cultured in a TSSBR system, which was beneficial to increase the proportion of Nitrosomonas in the microorganism of SBR2. It avoided the inhibition of the high organic loading to nitrification and C/N was not the limiting factor affecting the nitrogen removal efficiency. The rate of COD degradation and nitrification was improved, 8 kg COD/kg MLSS/d and 0.25 kg NH4+-N/kg MLSS/d, respectively. The final effluent COD concentration further reduced because the organic substrate that was left in SBR1 effluent was removed in SBR2. Consequently, when compared to a single SBR process, a two-stage SBR (TSSBR) not only improved the treatment efficiency, but also saved the operational cost.

Bioreactors↗
PubMed2004

Effect of drotrecogin alfa (activated) on human endothelial cell permeability and Rho kinase signaling.

OBJECTIVE: To explore whether the improvement in organ function and the vasoactive effect observed in the clinical studies of drotrecogin alfa (activated) (recombinant human activated protein C, rhAPC) in sepsis are a result of rhAPC's effect on endothelial cell (EC) permeability and modulation of the intracellular cytoskeleton via the Rho kinase signaling pathway. DESIGN: Findings regarding dose and duration of exposure to the drug with sequential addition of rhAPC and mediators (thrombin, histamine, interleukin-1 beta). SETTING: Research laboratory in a pharmaceutical company. SUBJECTS: Cultured primary human EC from different tissues and vascular beds. INTERVENTIONS: A monolayer of EC was incubated with either rhAPC, thrombin, histamine, or interleukin-1 beta alone or with rhAPC in combination with thrombin or interleukin-beta. The effect of rhAPC and mediators on EC permeability was monitored with measurement of electrical resistance. The effect on Rho kinase pathway signaling was monitored by the levels of phosphorylated myosin light chain and blockage with the Rho kinase specific inhibitor, Y27632. MEASUREMENTS AND MAIN RESULTS: Thrombin alone induced an early, concentration-dependent, and transient leakiness of EC. Interleukin-1 beta (0.5 ng/mL) induced an early, irreversible leakiness of EC. rhAPC (0.05-0.2 microg/mL, approximate median therapeutic blood levels) alone had no effect on EC permeability. rhAPC at > or=1 microg/mL induced an early EC leakage. rhAPC (0.19 microg/mL) attenuated the leakage induced by 0.5 ng/mL interleukin-1beta on microvascular EC derived from lung and skin and partially attenuated the leakage induced by 0.25 nM thrombin on human coronary arterial ECs. Levels of phosphorylated myosin light chain increased rapidly in human coronary arterial ECs when stimulated with thrombin or rhAPC (about 100-fold less potent) in a concentration-dependent manner via the Rho kinase signaling pathway. Short (5 mins) preconditioning of human coronary arterial ECs with 0.19 microg/mL rhAPC partially blocked the increase in phosphorylated myosin light chain levels induced by thrombin (0.06-0.2 nM). CONCLUSIONS: At concentrations exceeding physiologic and therapeutic levels, rhAPC increases EC permeability, an effect not seen at lower concentrations. The data suggest that interpretation of published in vitro and in vivo data of rhAPC and EC permeability should take into consideration the concentrations of rhAPC used or achieved. Other preliminary novel observations suggest that studying the effects of rhAPC on EC permeability and intracellular cytoskeletal organization may provide understanding of the effect of rhAPC on EC function.

Anti-Infective Agents↗
PubMed2004

[Experimental study on the antiviral mechanism of Ceratostigma willmattianum against herpes simplex virus type 1 in vitro].

OBJECTIVE: To study the antiviral effect and mechanisms of the liquid extract from Ceratostigma willmattianum against herpes simplex virus type 1 (HSV-1) in vitro. METHOD: C. willmattianum in various concentration was applied to different steps of HSV-1 replication cycle. 50% Tissue culture infective dose (TCID50), cytopathic effect (CPE), MTT staining method, dot blotting and Northern blotting analysis were used to estimate index of antiviral activity. RESULT: 50% Toxic concentration (TC50) was 1077 mg x L(-1), IC50 29.46 mg x L(-1) and therapeutic index (TI) 36.56 in C. willmattianum. TC50 330 mg x L(-1), 50% Inhibiting concentration (IC50) 9.12 mg x L(-1) and TI 36.18 in ACV by MTT staining method. The liquid extract from C. willmattianum had remarkable effect on inhibiting HSV-1 in vitro. Ceratostigma could interfere absorption of HSV-1 to Vero cells to prevent HSV-1 infectivity, inhibit HSV-1 gD DNA replication and HSV-1 gD mRNA expression. CONCLUSION: C. willmattianum possesses strong anti-HSV-1 activity in vitro. The antiviral mechanisms are related to inhibiting virus absorption, HSV-1 gD gene replication and HSV-1 gD gene transcription.

Animals↗
PubMed2003

Selective enhancement of multipotential hematopoietic progenitors in vitro and in vivo by IL-20.

In a search for novel growth factors, we discovered that human interleukin-20 (IL-20) enhanced colony formation by CD34+ multipotential progenitors. IL-20 had no effect on erythroid, granulocyte-macrophage, or megakaryocyte progenitors. IL-20 transgenic mice increased the numbers and cell cycling of multipotential but not other progenitors. IL-20 administration to normal mice significantly increased only multipotential progenitor cells, demonstrating that IL-20 significantly influences hematopoiesis, with specificity toward multipotential progenitors. This is the first cytokine with such specificity identified.

Animals↗
PubMed2003

A direct injection high-throughput liquid chromatography tandem mass spectrometry method for the determination of a new orally active alphavbeta3 antagonist in human urine and dialysate.

A generic high-throughput liquid chromatography (HTLC) tandem mass spectrometry (MS/MS) assay for the determination of compound I in human urine and dialysate (hemodialysis) was developed and validated. By using the HTLC on-line extraction technique, sample pretreatment was not necessary. The sample was directly injected onto a narrow bore large particle size extraction column (50 x 1.0 mm, 60 microm) where the sample matrix was rapidly washed away using a high flow rate (5 mL/min) aqueous mobile phase while analytes were retained. The analytes were subsequently eluted from the extraction column onto an analytical column using an organic-enriched mobile phase prior to mass spectrometric detection. The analytes were then eluted from the analytical column to the mass spectrometer for the determination. The linear dynamic range was 2.0-6000 ng/mL for the urine assay and 0.1-300 ng/mL for the dialysate assay. Intraday accuracy and precision were evaluated by analyzing five replicates of calibration standards at all concentrations used to construct the standard curve. For the urine assay, the precision (RSD%, n=5) ranged from 1.9 to 8.0% and the accuracy ranged from 87.8 to 105.2% of nominal value. For the dialysate assay, the precision (RSD%, n=5) ranged from 1.1 to 10.0% and the accuracy from 94.5 to 105.2% of nominal value. In-source fragmentation of the acyl glucuronide metabolite (compound III) did not interfere with the determination of parent compound I. The developed HTLC/MS/MS methodology was specific for compound I in the presence of compound III. Column life-time is increased and sample analysis time is decreased over traditional reversed-phase methods when direct injection assays for urine and dialysate are coupled with the technology of HTLC.

Chromatography, High Pressure Liquid↗
PubMed2003

Role of the serine-threonine kinase PAK-1 in myxoma virus replication.

Subversion or appropriation of cellular signal transduction pathways is a common strategy employed by viruses to promote an environment within infected cells that supports the viral replicative cycle. Using subsets of 3T3 murine fibroblasts previously shown to differ in their ability to support myxoma virus (MV) replication, we investigated the role of host serine-threonine kinases (STKs) as potential mediators of the permissive phenotype. Both permissive and nonpermissive 3T3 cells supported equivalent levels of virion binding, entry, and early virus gene expression, indicating that MV tropism in 3T3 cells was not determined by receptor-mediated entry. In contrast, late virus gene expression and viral DNA replication were selectively compromised in restrictive 3T3 cells. Addition of specific protein kinase inhibitors, many of which shared the ability to influence the activity of the STKs p21-activated kinase 1 (PAK-1) and Raf-1 attenuated MV replication in permissive 3T3 cells. Western blot detection of the phosphorylated forms of PAK-1 (Thr423) and Raf-1 (Ser338) confirmed activation of these kinases in permissive cells after MV infection or gamma interferon treatment, but the activated forms of both kinases were greatly reduced or absent in restrictive 3T3 cells. The biological significance of these activations was demonstrated by using the autoinhibitory domain of PAK-1 (amino acids 83 to 149), expression of which reduced the efficiency of MV infection in permissive 3T3 cells concurrent with a decrease in PAK-1 activation. In comparison, overexpression of a constitutively active PAK-1 (T423E) mutant increased MV replication in restrictive 3T3 cells. These observations suggest that induced signaling via cellular STKs may play important roles in determining the permissiveness of host cells to poxvirus infection.

3T3 Cells↗