ABSTRACT
BACKGROUND Since the epidemiological profile of drug-induced liver injury (DILI) in China, especially the western of China, it has rarely been studied. The aim of this study was to analyze the characteristics of DILI patients in a large tertiary teaching hospital at Chongqing, a municipality in western China. MATERIAL AND METHODS The medical records of hospitalized patients which diagnosed with DILI between January 2011 and December 2016 were searched retrospectively, and demographic, clinical data, and laboratory data were retrieved for analysis. RESULTS A total of 1811 patients had been diagnosed with DILI, accounting for 0.248% of the total admissions during the same period. Among the 1096 patients included in our analysis, DILI was caused by "medications" in 462 cases (42.15%), "herbs" in 391 cases (35.68%), and combined medications in 189 cases (17.24%). The profiles for each etiology were distinctive for age, sex, clinical features, laboratory features, and types and severity of DILI. CONCLUSIONS Our study provides a systematic etiological profile of DILI in Chinese patients, which can represent references for prevention, diagnosis and treatment, supporting and promoting efforts to ease the burden of this liver disease in China.
Subject(s)
Chemical and Drug Induced Liver Injury/epidemiology , Chemical and Drug Induced Liver Injury/etiology , Adolescent , Adult , Aged , Aged, 80 and over , Chemical and Drug Induced Liver Injury/diagnosis , Chemical and Drug Induced Liver Injury/pathology , Child , Child, Preschool , China/epidemiology , Cost of Illness , Drug Therapy, Combination/adverse effects , Female , Hospitals, Teaching/statistics & numerical data , Humans , Liver/drug effects , Liver/pathology , Male , Middle Aged , Patient Admission/statistics & numerical data , Retrospective Studies , Severity of Illness Index , Tertiary Care Centers/statistics & numerical data , Young AdultABSTRACT
OBJECTIVE: To explore the metabolic characteristics and metabolic markers of WBC-depleted RBCs in MAP preservation solution and to analyzed the metabolic profile of RBC in MAP preservation solution by using metabolomics. METHODS: The changes of metabolitcs in 10 U WBC-depleted RBC suspension at 3-different storage period (D 0, D 14 and D 35) were detected by using the UPLC-MS/MS, the charaeteristic ions and metabolic markers of RBC stored in preservation sblution for 0 d, 14 d and 35 days were analyzed by using the principal component analysis(PCA). RESULTS: The number of characteristic ions in RBC and supernatant extracts detected during the initial, middle and final storage could be clearly distinguiseed. The 5 metabolism-related substamces such as lact-c acid, nicotinamide, glucose, 5-htdroxyproline and malic acid showed statistically significant difference in 3 storage period. CONCLUSION: The UPLC-MS/MS method combined with statistical analysis of multivariate data can be used to study the metabolic characteristics of RBC, the different metabolites of RBC in different storage stages can be used as the potential markers for evaluation of guality of RBC in storag period. The results of this study provide a basis for studing the RBC guality changes in storage period.
Subject(s)
Blood Preservation , Tandem Mass Spectrometry , Chromatography, Liquid , Erythrocytes , MetabolomeABSTRACT
BACKGROUND/PURPOSE: Subminimal inhibitory concentration (sub-MIC) of antibiotics can modify the phenotype of biofilm formation in bacteria. However, the relationship between resistance phenotypes, genotypes, and the biofilm formation phenotype in response to sub-MIC antibiotics remains unclear. METHODS: Here, we collected 96 clinical isolates of Staphylococcus epidermidis (S. epidermidis) and investigated the erythromycin (ERY) susceptibility, the biofilm formation in respond to sub-MIC ERY, the presence and transcription expression of erm genes. Serial passage of induction resistance was used against ERY-susceptible isolates and biofilm formation in response to their new sub-MIC ERY was determined. RESULTS: The incidence of biofilm phenotype modification in ERY-resistant isolates was significantly higher than that of ERY-susceptible isolates [27/85 (31.8%) vs. 0/11 (0%), p = 0.031]. Yet, ERY-susceptible isolates displayed the phenomenon of biofilm phenotype modification (7/11), after induction of resistance to ERY. The ermC gene was absolutely dominant among the three macrolide resistant genes including erm (A, B, C) [6/96 (6.2%), 6/96 (6.2%), and 91/96 (94.8%), respectively]. With statistic stratification analysis, a linear and positive correlation was identified between the two factors in the biofilm-enhanced strains, a linear and negative correlation in biofilm-inhibited strains, and a weakly positive correlation in biofilm-unaffected strains (R(2) = 0.4992, 0.3686, and 0.0512, respectively). CONCLUSION: The results suggest that the ERY resistance phenotype and the transcription expression of ermC gene could be considered as important signs to estimate whether the biofilm formation phenotype in S. epidermidis clinical isolates can be easily affected by sub-MIC ERY.
Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Drug Resistance, Bacterial , Erythromycin/pharmacology , Staphylococcus epidermidis/drug effects , Staphylococcus epidermidis/physiology , Gene Expression Profiling , Genes, Bacterial , Humans , Microbial Sensitivity Tests , Staphylococcal Infections/microbiology , Staphylococcus epidermidis/isolation & purification , tRNA Methyltransferases/geneticsABSTRACT
To investigate the effects of subminimum inhibitory concentrations of cephalosporins on bacterial biofilm formation, the biofilm production of 52 Escherichia (E.) coli strains was examined following treatment with cephalosporin compounds at 1/4 minimum inhibitory concentrations (MICs). Ceftazidime (CAZ) inhibited biofilm formation in seven isolates, while cefoperazone (CFP) enhanced biofilm formation in 18 isolates. Biofilm formation of E. coli E42 was inhibited by CAZ and induced by CFP. Therefore, using reverse transcriptionpolymerase chain reaction, the expression of the biofilmmodulating genes of this isolate was investigated. To monitor the production of the autoinducer of quorum sensing in E. coli, autoinducer2 (AI2) production was detected by measuring the bioluminescence response of Vibrio harveyi BB170. Antisense oligonucleotides (ASODNs) targeting Sribosylhomocysteine lyase (luxS) inhibited the expression of the luxS gene in E. coli. CAZ at 1/4 MIC reduced luxS mRNA levels and the production of AI2, whereas CFP at 1/4 MIC had the opposite effect. ASODNs targeting luxS significantly decreased the aforementioned inhibitory effects of CAZ and the induction effects of CFP on E. coli biofilm formation. Therefore, biofilm formation by the E. coli clinical isolate E42 was evoked by CFP but attenuated by CAZ at subMICs, via a luxS/AI2based quorum sensing system.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , Biofilms/drug effects , Carbon-Sulfur Lyases/metabolism , Cefoperazone/pharmacology , Ceftazidime/pharmacology , Escherichia coli Proteins/metabolism , Escherichia coli/physiology , Bacterial Proteins/genetics , Carbon-Sulfur Lyases/genetics , Escherichia coli/drug effects , Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Escherichia coli Proteins/genetics , Gene Expression/drug effects , Gene Expression Regulation, Bacterial/drug effects , Genes, Bacterial , Humans , Microbial Sensitivity Tests , Quorum Sensing/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
Acinetobacter baumannii is an important nosocomial pathogen able to cause severe infections in an intensive care unit (ICU). However, there is a lack of analysis regarding the epidemiology and resistance of A. baumannii in respiratory department ICUs. In this study, clinical isolates were collected from the respiratory department ICU of Southwest Hospital from January 2009 to December 2010, and the social and demographic information of the patients from whom the isolates were taken was obtained from the Southwest Hospital information system. The minimal inhibitory concentration (MIC) of the isolates was determined by the agar dilution method. The carbapenemase-encoding resistance genes of these isolates were amplified using PCR. The clonal relationship of isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Forty-six isolates were collected from the respiratory department ICU, and the antibiotics minocycline and quinolone had higher drug sensitivity against these isolates. The OXA-51, OXA-23, and IMP-4 genes were present at rates of 100% (46/46), 67.4% (31/46), and 41.3% (19/46), respectively. Forty-six isolates had 12 different PFGE genotypes. The results above suggested that the hospital environment and patients contributed to nosocomial infections, and the spread of resistance genes in the hospital was common.
Subject(s)
Acinetobacter Infections/epidemiology , Acinetobacter baumannii/genetics , Acinetobacter baumannii/isolation & purification , Drug Resistance, Multiple, Bacterial/genetics , Acinetobacter Infections/drug therapy , Acinetobacter Infections/microbiology , Acinetobacter baumannii/drug effects , Adolescent , Adult , Aged , Anti-Bacterial Agents/therapeutic use , Bacterial Proteins/genetics , Child , Child, Preschool , China/epidemiology , Cross Infection/epidemiology , Drug Resistance, Multiple, Bacterial/drug effects , Electrophoresis, Gel, Pulsed-Field/methods , Female , Hospitals , Humans , Infant , Infant, Newborn , Intensive Care Units , Male , Microbial Sensitivity Tests/methods , Middle Aged , Minocycline/therapeutic use , Quinolones/therapeutic use , Young Adult , beta-Lactamases/geneticsABSTRACT
Although Pseudomonas aeruginosa is not typically susceptible to azithromycin (AZM) in in vitro tests, AZM improves the clinical outcome in patients with chronic respiratory infections, in which both the modulation of the host immune system and of bacterial virulence by AZM are thought to play an important role. However, there is currently little direct evidence showing the impact of bacteria pretreated with AZM on epithelial cells, which represents the first barrier to infecting P. aeruginosa. In this study, we pretreated P. aeruginosa with AZM and subsequently infected human bronchial epithelial cells (HBEs) in the absence of AZM. The results showed that AZM-pretreated P. aeruginosa (PAO1 and six different clinical isolates) significantly stimulated HBE cells to release IL-8, a crucial pro-inflammatory cytokine. This effect was not observed in a P. aeruginosa PAO1 mutant strain unable to produce the type III secretion system effector gene pcrV (strain PW4017). Our results suggest that AZM-pretreated P. aeruginosa could indirectly exacerbate pro-inflammation by inducing IL-8 production in HBEs.
Subject(s)
Anti-Bacterial Agents/metabolism , Azithromycin/metabolism , Epithelial Cells/immunology , Epithelial Cells/microbiology , Interleukin-8/metabolism , Pseudomonas aeruginosa/immunology , Pseudomonas aeruginosa/metabolism , Cell Line , HumansABSTRACT
The objective of this study was to evaluate the efficacy and safety of a traditional Chinese medicine, Fufang Xuelian Burn Ointment (FXBO), to treat superficial and deep second-degree burn wounds. A four-center, randomized, controlled, and prospective study was conducted. Overall, 240 patients with either superficial or deep second-degree burn wounds were enrolled consecutively in this study. Patients who were randomly assigned to the control group (superficial: 72, deep: 48) underwent common burn wound therapy, whereas those randomized to the treatment group (superficial: 72, deep: 48) received common burn wound therapy plus topical FXBO. The healing rate, healing time, effective rate, and safety data were compared between the two groups. The baseline characteristics were comparable for the two groups. The healing rate was 94.79(±7.50) in the control group and 98.60(±5.69) in the FXBO group after 14 days for patients with superficial second-degree burn wounds (P = 0.000), and 95.17(±9.68) versus 97.44(±9.81) at 28 for deep second-degree burn wounds (P = 0.025). The median healing time in the FXBO group were 9 and 21 days for superficial and deep second-degree burns, respectively, compared to 10.5 and 22.5 days, respectively, in control group (P(superficial) = 0.000 and P(deep) = 0.009). The results of the effective rate showed that comprehensive efficacy of the FXBO group was improved compared to the control group for either superficial or deep second-degree burns (P(superficial) = 0.035 and P deep = 0.003). There were no reported drug-related adverse events in both groups. Therefore, FXBO was well tolerated and more effective than control group for treating superficial and deep second-degree burn wounds.
Subject(s)
Burns/drug therapy , Drugs, Chinese Herbal/therapeutic use , Adolescent , Adult , Aged , Burns/physiopathology , Drugs, Chinese Herbal/adverse effects , Female , Humans , Male , Middle Aged , Ointments , Prospective Studies , Safety , Treatment Outcome , Wound Healing/drug effects , Young AdultABSTRACT
The carbapenemases have recently emerged as molecules implicated in one of the most feared bacterial resistance mechanisms because of their ability to hydrolyze virtually all lactamase agents and their highly mobile genes. This study aimed to investigate the prevalence of carbapenemase and antimicrobial susceptibilities of Pseudomonas aeruginosa isolated from burn patients in Chongqing, China. Antimicrobial susceptibility of 111 isolates was determined by the disc agar diffusion test and the agar dilution method. Random Amplification of Polymorphic DNA polymerase chain reaction analysis revealed 111 P. aeruginosa 42 genotypes. Carbapenemase genes were amplified by polymerase chain reaction and the sequence verified by blast. Ninety-three of 111 (83.8%) isolates were resistant to imipenem; all of them had developed multidrug resistance and exhibited higher resistant rates compared with the imipenem-susceptible Pseudomonas. Ciprofloxacin was the most effective antipseudomonal agent. Thirty-three of the isolates were identified to contain the metallo-ß-lactamase blaIMP-4 gene and belong to different Random Amplification of Polymorphic DNA polymerase chain reactiongenotypes. In conclusion, the high prevalence of multidrug resistance (94.6%) and the production of blaIMP-4 genes in P. aeruginosa isolates in burn patients highlight the necessity of considering these issues in burn hospitals.
Subject(s)
Bacterial Proteins/genetics , Burns/microbiology , Pseudomonas aeruginosa/genetics , beta-Lactamases/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Anti-Bacterial Agents/therapeutic use , Burn Units , Child , Child, Preschool , China , DNA, Bacterial/genetics , Drug Resistance, Multiple, Bacterial/genetics , Female , Genotype , Humans , Male , Middle Aged , Polymerase Chain Reaction , Pseudomonas aeruginosa/isolation & purification , Young AdultABSTRACT
The looped host defense peptide CLP-19 is derived from a highly functional core region of the Limulus anti-LPS factor and exerts robust anti-LPS activity by directly interacting with LPS in the extracellular space. We previously showed that prophylactic administration of CLP-19 even 20 h prior to LPS challenge might significantly increase the survival rate in a lethal endotoxin shock mouse model. Such an effect may be associated with immune regulation of CLP-19. To investigate the underlying mechanisms, peptide affinity chromatography, immunofluorescence, and Western blotting procedures were used to identify α- and ß-tubulin as direct and specific binding partners of CLP-19 in the mouse macrophage cell line RAW 264.7. Bioinformatic analysis using the AutoDock Vina molecular docking and PyMOL molecular graphics system predicted that CLP-19 would bind to the functional residues of both α- and ß-tubulin and would be located within the groove of microtubules. Tubulin polymerization assay revealed that CLP-19 might induce polymerization of microtubules and prevent depolymerization. The immunoregulatory effect of CLP-19 involving microtubules was investigated by flow cytometry, immunofluorescence, and Western blotting, which showed that CLP-19 prophylactic treatment of RAW 264.7 cells significantly inhibited LPS-induced surface expression of TLR4. Taken together, these results suggest that CLP-19 binding to microtubules disrupts the dynamic equilibrium of microtubules, reducing the efficacy of microtubule-dependent vesicular transport that would otherwise translocate TLR4 from the endoplasmic reticulum to the cell surface.
Subject(s)
Antimicrobial Cationic Peptides/metabolism , Arthropod Proteins/metabolism , Macrophages/metabolism , Microtubules/metabolism , Protein Transport/physiology , Toll-Like Receptor 4/metabolism , Animals , Antimicrobial Cationic Peptides/chemistry , Antimicrobial Cationic Peptides/immunology , Arthropod Proteins/chemistry , Arthropod Proteins/immunology , Blotting, Western , Cell Line , Cell Membrane/immunology , Cell Membrane/metabolism , Chromatography, Affinity , Flow Cytometry , Fluorescent Antibody Technique , Macrophages/immunology , Membrane Proteins/immunology , Membrane Proteins/metabolism , Mice , Microtubules/immunology , Peptides, Cyclic/chemistry , Peptides, Cyclic/immunology , Peptides, Cyclic/metabolism , Toll-Like Receptor 4/immunology , Tubulin/immunology , Tubulin/metabolismABSTRACT
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 hyperoside's 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.
Subject(s)
Antioxidants/pharmacology , Cytoprotection , Heme Oxygenase-1/genetics , Hydrogen Peroxide/antagonists & inhibitors , Intracellular Signaling Peptides and Proteins/metabolism , NF-E2-Related Factor 2/metabolism , Quercetin/analogs & derivatives , Transcriptional Activation , Active Transport, Cell Nucleus/drug effects , Cell Nucleus/metabolism , Extracellular Signal-Regulated MAP Kinases/metabolism , Humans , Hydrogen Peroxide/toxicity , Kelch-Like ECH-Associated Protein 1 , Oxidative Stress/genetics , Quercetin/pharmacology , Response Elements , Signal Transduction , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
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.
Subject(s)
Anti-Bacterial Agents/administration & dosage , Biofilms/drug effects , Biofilms/growth & development , Genes, Bacterial , Macrolides/administration & dosage , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Staphylococcus epidermidis/drug effects , Staphylococcus epidermidis/genetics , Base Sequence , DNA Primers/genetics , Dose-Response Relationship, Drug , Erythromycin/administration & dosage , Gene Expression/drug effects , Humans , In Vitro Techniques , Microscopy, Electron, Scanning , Reverse Transcriptase Polymerase Chain Reaction , Staphylococcus epidermidis/isolation & purification , Staphylococcus epidermidis/physiologyABSTRACT
OBJECTIVE: To investigate the role of REMP2 derived from limulus anti-lipopolysaccharide factor in neutralizing endotoxin in vitro and its antibacterial activity. METHODS: (1) REMP2 and PMB in the concentrations of 100.00, 10.00, 1.00, 0.10, 0.01 micromol/L were respectively mixed with LPS (lEU/mL), with PMB as positive control. The LPS concentrations in different specimens were determined by routine method, and the neutralizing percentage was respectively calculated. (2) After adding isotonic saline (NS), the final concentrations of REMP2 and PMB were 10, 20, 40, 80 micromol/L, and the concentration of LPS was 100 microg/L. The murine monocytic macrophages were stimulated with LPS, then cultured with REMP2 and PMB, with NS in culture as negative control. The content of tumor necrosis factor (TNF)-alpha was determined by ELISA kit. (3) The morphologic changes of Escherichia coli. was observed under electron microscope at 10, 20 and 40 minutes after addition of REMP2 to Escherichia coli suspension (with terminal concentration of REMP2 at 40 micromol/L). RESULTS: There were no significant difference in endotoxin-neutralizing percentages between PMB and REMP2 in concentrations of 0.10, 10.00, 100.00 micromol/L (P > 0.05). The contents of TNF-alpha were 1175 +/- 162, 859 +/- 122, 645 +/- 142, 489 +/- 102 ng/L, respectively,after treatment of 10, 20, 40, 80 micromol/L REMP2, which were obviously lower than that of NS (3463 +/- 218 ng/L, P < 0.01). Under transmission electron microscope, the outer and interior membranes of Escherichia coli were obscure and rough, bacterial bodies were swollen with vacuoles in cytoplasm after treatment with REMP2. CONCLUSION: REMP2 has ability of neutralizing endotoxin and also antibacterial activity.
Subject(s)
Anti-Bacterial Agents/pharmacology , Invertebrate Hormones/pharmacology , Peptide Fragments/pharmacology , Animals , Antimicrobial Cationic Peptides , Arthropod Proteins , Cells, Cultured , Escherichia coli/drug effects , Escherichia coli/metabolism , Limulus Test , Lipopolysaccharides/antagonists & inhibitors , Macrophages/metabolism , Mice , Tumor Necrosis Factor-alpha/metabolismABSTRACT
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.
Subject(s)
Endotoxemia/prevention & control , Invertebrate Hormones/chemistry , Lipopolysaccharides/metabolism , Macrophages/metabolism , Peptides, Cyclic/metabolism , Peptides, Cyclic/pharmacology , Tumor Necrosis Factor-alpha/metabolism , Animals , Antimicrobial Cationic Peptides , Arthropod Proteins , Cell Line , Disease Models, Animal , Horseshoe Crabs/metabolism , Limulus Test , Macrophages/drug effects , Mice , Mice, Inbred Strains , Peptides, Cyclic/chemical synthesis , Peptides, Cyclic/chemistry , Tumor Necrosis Factor-alpha/bloodABSTRACT
OBJECTIVE: To reproduce a Kunming murine endotoxin shock model suitable for the anti-endotoxin pharmaceutical research. METHODS: Kunming mice were challenged with an intraperitoneal (i. p.) injection of different doses of D-galactosamine (D-Gal) and endotoxin (LPS) and divided into 10 groups: i.e, group 1 [with injection of isotonic saline solution (NS) and LPS]; group 2 (with injection of NS and 90mg/kg LPS), group 3 (with injection of NS and 500mg/kg D-Gal), group 4 (with injection of 500mg/kgD-Gal and 25 microg /kg LPS), group 5 (with injection of 500mg/kg D-Gal and 50 microg/kg LPS), group 6(with injection of 500mg/kg D-Gal and 250 microg/kg LPS), group 7( with injection of NS and 600mg/ kg D-Gal), group 8 (with injection of 600mg/kg D-Gal and 10 microg/kg LPS), group 9( with injection of 600mg/kg D-Gal and 25 microg/kg LPS), group 10 (with injection of 600mg/kg D-Gal and 50 microg/kg LPS). The death of the mice were observed and the mortality rate was recorded at 48 post-injection hour (PIH). The dose of D-Gal and LPS which caused 100% lethality was chosen for the subsequent experiment to serve as control group (with injection of NS and 600mg/kg D-Gal), LPS group (with injection of 600mg/kg D-Gal and 580mg/kg LPS for later experiment). The venous blood of the mice were collected for the detection of serum content of TNF-alpha with ELISA method at 30, 75 and 120 post-injection minutes (PIM). The tissues of lung, liver, intestine were also harvested at 5 PIH for the pathological examination. RESULTS: The lethality of mice was 100% in the groups 2, 6 and 10 (P < 0.01). The serum content of TNF-alpha was maintained in a low level in control group, but it increased remarkably in LPS group, and it reached peak at 75 PIM (6365 +/- 2087ng/L, P < 0.01). Obvious inflammatory reaction was observed in the lung, liver and intestine in LPS group, while only mild inflammatory reaction was observed in liver in control group. CONCLUSION: The Kunming mice showed signs of endotoxin shock after D-galactosamine presensitizing and endotoxin challenge, and it is suitable for anti-endotoxin pharmaceutical research.
Subject(s)
Disease Models, Animal , Galactosamine/adverse effects , Shock, Septic/chemically induced , Animals , Female , Male , Mice , Mice, Inbred Strains , Serum/chemistry , Tumor Necrosis Factor-alpha/metabolismABSTRACT
OBJECTIVE: To investigate the drug resistance of flavobacterium and its ability to produce BLA (beta-lactamases) and ESBLs (Extended-spectrum beta-lactamases). METHODS: The production of BLA and ESBLs from 6 clinical isolated flavobacterium strains was determined by nitrocefin disc test and double-disc synergy method, respectively. The antibiotic susceptibilities of the strains were determined by Kirby-Bauer disc diffusion test and the agar dilution method and the MIC was assessed. RESULTS: All the six flavobacteria were BLA-producing strains and more than 80% of them were ESBLs-producing, and they were highly resistant to beta-lactamase antibiotics (MIC 32 - 256 mg/L), but susceptible to fluoroquinolones and cephalosporin with beta-lactamase inhibitors (MIC 0.125 - 8 mg/L). CONCLUSION: Most of the flavobacteria in nosocomial infections were beta-lactamase-producing and were highly resistant to beta-lactamase antibiotics. Fluoroquinolones and beta-lactamase antibiotics with lactamase inhibitors should be the first choice for the management of infection caused by flavobacterium.
Subject(s)
Drug Resistance, Bacterial , Flavobacterium/drug effects , Flavobacterium/enzymology , Membrane Proteins/analysis , Ribosomal Proteins/analysis , Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial/drug effects , Humans , Membrane Proteins/metabolism , Microbial Sensitivity Tests , Ribosomal Proteins/metabolismABSTRACT
OBJECTIVE: To observe different degrees of intra-abdominal pressure and different duration on the intestinal permeability and endotoxin/bacteria translocation in rabbit model, so as to explore the mechanism of the development of abdominal compartment syndrome (ACS) and MODS. METHODS: Rabbit model of intra-abdominal hypertension was established by injection of gaseous nitrogen into the peritoneal cavity. Thirty-nine New Zealand white rabbits were employed in the study. The change in intestinal permeability was determined by fluorescein isothiocyanate dextran (FITC-D) and two kinds of molecular probes of type II horseradish peroxidase (HRP-II). The effects of intra-abdominal hypertension on the endotoxin/bacteria translocation were also detected. RESULTS: The contents of FITC-D and HRP-II in portal veins increased evidently (P < 0.01) when intra-abdominal pressure (IAP) was higher than 20 mmHg. The endotoxin (ET) content in portal vein in rabbits with IAP of 10 mmHg for 1, 2 and 4 hours exhibited no difference compared with that in normal control, while the ET content increased obviously after 1 hour with IAP of 20 mmHg and increased thereafter along with the prolongation of IAP, and increase in pressure. The bacterial translocation rates were 33.3%, 66.7% and 100% when IAP was maintained at 20 mmHg for 1, 2 and 4 hours, respectively, and there was evidence of bacterial translocation to the liver. The rate of bacterial translocation to intestinal mesenteric lymph nodes was 100% when IAP was 30 mmHg for 1 and 2 hours. There was no bacterial translocation to the spleen in all experimental rabbits. CONCLUSION: Intestinal mucosal permeability increased significantly with increased endotoxin content in portal vein when IAP was higher than 20 mmHg. At the sane time, the bacteria could be translocate to intestinal mesenteric lymph nodes and liver, which might be constitute one of the important factors leading to the development of ACS and MODS.
Subject(s)
Abdomen/microbiology , Abdomen/physiopathology , Bacterial Translocation , Endotoxins/blood , Intestines/physiopathology , Animals , Colony Count, Microbial , Compartment Syndromes/etiology , Female , Male , Multiple Organ Failure/etiology , Permeability , RabbitsABSTRACT
AIM: To investigate the relationship between lymphocyte apoptosis in peripheral blood, spleen and mesenteric lymph nodes(MLN) and endotoxin translocation after thermal injury in rats. METHODS: In a Wistar rat model inflicted with 30% TBSA III degree scalding, serum LPS levels in portal vein and vena cava were quantified by tachypleus amebocyte lysate (TAL) technique. The analysis of peripheral blood lymphocyte was employed in in situ Cell Death Detection Kit and evaluated by flow cytometry. Apoptotic lymphocytes in paraffin-embedded spleen and MLN sections were examined by histologic analysis, in situ deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) and peroxidase (POD) staining. The images were taken by Cooldccd camera system, and the count and optical density value (transmission light) of apoptotic lymphocytes were analyzed with software Spot and Imagine proplus 4.10a(IPP4.10a). RESULTS: In the period of 3 to 48 postburn hours (PBHs) serum LPS level (x10(3) EU.L(-1)) in portal vein (2.11+/-0.02, 5.66+/-0.20, 3.70+/-0.22, 2.56+/-0.28, 0.90+/-0.11) was higher than that in vena cava (0.63+/-0.01, 1.53+/-0.18, 0.83+/-0.32, 0.52+/-0.12, 0.23+/-0.02, P<0.01), but both increased sharply in postburn rats (P<0.01) and reached a peak at 6 PBH. Analysis of apoptotic lymphocytes showed that the proportion (%) of postburn apoptotic cells was much higher than that in healthy rats (8.34+/-1.53, 8.13+/-1.81, 20.77+/-3.94, 23.90+/-3.92, 11.23+/-1.35 and 13.26+/-2.09 at 3, 6, 12, 24, 48 and 72 PBH, respectively, vs 3.99+/-1.72, P<0.01), especially after 6 PBH. The concentrations of lymphocytic apoptosis at 12 and 24 PBH were markedly higher than that at other time points. Meantime, few apoptotic lymphocytes were found in normal MLN, but increased postburn obviously (3+/-1 vs 546+/-83, 285+/-39, 149+/-30, 58+/-10, 36+/-11 and 33+/-9 in turn, P<0.01), especially at 3 PBH, whereas apoptotic lymphocytes were concentrated in splenic cortex before the burn and decreased obviously during 72 PBHs (499+/-186 vs 12+/-8, 19+/-15, 12+/-7, 100+/-15, 123+/-25 and 226+/-26 in turn, P<0.01) though a slight rise was found in the medulla after 24 PBH. Optical density of apoptotic lymphocytes was significantly reduced in spleen in the 24 PBHs and raised in MLN during 48 PBHs than that prior to the burn, respectively. CONCLUSION: Gut-origin LPS is a major cause of endotoxemia taken place early in rats following severe thermal injury and could induce extensive lymphocyte apoptosis in blood and MLN, which suggests an immunosuppression state could follow the initial injury and favores a septic state based on apoptotic mechanism.