Based on the interleukin-17(IL-17) signaling pathway, this study explores the effect and mechanism of Bufei Decoction on Klebsiella pneumoniae pneumonia in rats. SD rats were randomly divided into the control group, model group, Bufei Decoction low-dose group(6.68 g·kg~(-1)·d~(-1)), Bufei Decoction high-dose group(13.36 g·kg~(-1)·d~(-1)), and dexamethasone group(1.04 mg·kg~(-1)·d~(-1)), with 10 rats in each group. A pneumonia model was established by tracheal drip injection of K. pneumoniae. After successful model establishment, the improvement in lung tissue damage was observed following drug administration. Core targets and signaling pathways were screened using transcriptomics techniques. Real-time fluorescence quantitative polymerase chain reaction was used to detect the mRNA expression of core targets interleukin-6(IL-6), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and chemokine CXC ligand 6(CXCL6). Western blot was used to assess key proteins in the IL-17 signaling pathway, including interleukin-17A(IL-17A), nuclear transcription factor-κB activator 1(Act1), tumor necrosis factor receptor-associated factor 6(TRAF6), and downstream phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK), and phosphorylated nuclear factor-κB p65(p-NF-κB p65). Apoptosis of lung tissue cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL). The results showed that, compared with the control group, the model group exhibited significant pathological damage in lung tissue. The mRNA expression of IL-6, IL-1β, TNF-α, and CXCL6, as well as the protein levels of IL-17A, Act1, TRAF6, p-p38 MAPK/p38 MAPK, and p-NF-κB p65/NF-κB p65, were significantly increased, and the number of apoptotic cells was notably higher, indicating successful model establishment. Compared with the model group, both low-and high-dose groups of Bufei Decoction showed reduced pathological damage in lung tissue. The mRNA expression levels of IL-6, IL-1β, TNF-α, and CXCL6, and the protein levels of IL-17A, Act1, TRAF6, p-p38 MAPK/p38 MAPK, and p-NF-κB p65/NF-κB p65, were significantly decreased, with a significant reduction in apoptotic cells in the high-dose group. In conclusion, Bufei Decoction can effectively improve lung tissue damage and reduce inflammation in rats with K. pneumoniae. The mechanism may involve the regulation of the IL-17 signaling pathway and the reduction of apoptosis.
Animals ; Interleukin-17/metabolism* ; Drugs, Chinese Herbal/administration & dosage* ; Rats, Sprague-Dawley ; Signal Transduction/drug effects* ; Rats ; Male ; Klebsiella pneumoniae/physiology* ; Klebsiella Infections/immunology* ; Humans ; Lung/drug effects*

LysR-type transcriptional regulators are involved in the regulation of numerous cellular metabolic processes in Klebsiella pneumoniae, leading to severe infection. Earlier, we found a novel LysR family gene, named kp05372, in a strain of K. pneumoniae (designated GPKP) isolated from forest musk deer. To study the function of this gene in relation to the biological characteristics of GPKP, we used the suicide plasmid and conjugative transfer methods to construct deletion mutant strain GPKP-Δkp05372; moreover, we also constructed the GPKP-Δkp05372+ complemented strain. The role of this gene was determined by comparing the following characteristics of three strains: growth curves, biofilm formation, drug resistance, stress resistance, median lethal dose (LD₅₀), organ colonization ability, and the histopathology of GPKP. Real-time polymerase chain reaction (RT-PCR) was used to test the expression level of seven genes upstream of kp05372. There was no significant difference in the growth rates when comparing the three bacterial strains, and no significant difference was recorded at different osmotic pressures, temperatures, salt contents, or hydrogen peroxide concentrations. The GPKP-Δkp05372 mutant formed a weak biofilm, and the other two strains formed medium biofilm. The drug resistance of the GPKP-Δkp05372 mutant toward cephalothin, cotrimoxazole, and polymyxin B was changed. The acid tolerance of the deletion strain was stronger than that of the other two strains. The LD₅₀ values of the wild-type and complemented strains were 174-fold and 77-fold higher than that of the GPKP-Δkp05372 mutant, respectively. The colonization ability of the GPKP-Δkp05372 mutant in the heart, liver, spleen, kidney, and intestine was the weakest. The three strains caused different histopathological changes in the liver and lungs. In the GPKP-Δkp05372 mutant, the relative expression levels of kp05374 and kp05379 were increased to 1.32-fold and 1.42-fold, respectively, while the level of kp05378 was decreased by 42%. Overall, the deletion of kp05372 gene leads to changes in the following: drug resistance and acid tolerance; decreases in virulence, biofilm formation, and colonization ability of GPKP; and regulation of the upstream region of adjacent genes.
Animals ; Bacterial Proteins/physiology* ; Biofilms ; Deer/microbiology* ; Drug Resistance, Bacterial ; Female ; Klebsiella Infections/pathology* ; Klebsiella pneumoniae/growth & development* ; Male ; Mice ; Transcription Factors/physiology*

Aortic Rupture ; diagnostic imaging ; microbiology ; surgery ; Blood Pressure ; physiology ; Humans ; Klebsiella pneumoniae ; pathogenicity ; Male ; Middle Aged

OBJECTIVETo characterize carbapenem (CPM)-non-susceptible Klebsiella pneumoniae (K. pneumoniae) and carbape-nemase produced by these strains isolated from Beijing Children's Hospital based on a five-year surveillance.

METHODSThe Minimal Inhibition Concentration values for 15 antibiotics were assessed using the Phonix100 compact system. PCR amplification and DNA sequencing were used to detect genes encoding carbapenemases. WHONET 5.6 was finally used for resistance analysis.

RESULTSIn total, 179 strains of CPM-non-susceptible K. pneumoniae were isolated from January, 2010 to December, 2014. The rates of non-susceptible to imipenem and meropenem were 95.0% and 95.6%, respectively. In the 179 strains, 95 (53.1%) strains carried the blaIMP gene, and IMP-4 and IMP-8 were detected in 92 (96.8%) and 3 (3.2%) IMP-producing isolates, respectively. 65 (36.3%) strains carried the blaNDM-1 gene. 6 (3.4%) strains carried the blaKPC gene, and KPC-2 were detected in 6 KPC-producing isolates. In addition, New Delhi-Metallo-1 (NDM-1) producing isolates increased from 7.1% to 63.0% in five years and IMP-4 producing isolates decreased from 75.0% to 28.3%.

CONCLUSIONHigh frequencies of multiple resistances to antibiotics were observed in the CPM-non-susceptible K. pneumoniae strains isolated from Beijing Children's Hospital. The production of IMP-4 and NDM-1 metallo-β-lactamases appears to be an important mechanism for CPM-non- susceptible in K. pneumoniae.

Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Child ; China ; epidemiology ; Drug Resistance ; Gene Expression Regulation, Bacterial ; physiology ; Gene Expression Regulation, Enzymologic ; physiology ; Hospitals, Pediatric ; Humans ; Klebsiella Infections ; epidemiology ; microbiology ; Klebsiella pneumoniae ; drug effects ; enzymology ; genetics ; Microbial Sensitivity Tests ; Population Surveillance ; Time Factors ; beta-Lactamases ; genetics ; metabolism

Objective To explore the effects of Scutellaria baicalensis on activity and biofilm formation of Klebsiella pneumonia (Kp).Methods The broth and agar dilution Methods were carried out to determine minimum inhibitory concentration and minimum bactericidal concentration of Scutellaria baicalensis for TW518. VITEK-32 system was used to assay TW518 susceptibility to antibiotics. Kp biofilms were formed in vitro and stained with BacLight Live/Dead stain. The class integron geneⅠ1 mRNA expression was analyzed with RT-PCR.Results The minimum inhibitory concentration of Scutellaria baicalensis on TW518 identified as a Kp colony was 32 mg/ml, and minimum bactericidal concentration was 64 mg/ml. Scutellaria baicalensis and broad-spectrum penicillin, cephalosporin, quinolones, or beta-lactamase had synergistic bactericidal effects. Biofilm formation activity of Kp treated with Scutellaria baicalensis was significantly lower than that of the control group. And class integron geneⅠ1 mRNA expression of TW518 was significantly inhibited by Scutellaria baicalensis.Conclusions Scutellaria baicalensis has sterilization effect on Kp, and Scutellaria baicalensis could effectively inhibit Kp biofilm formation with prolonged treatment. Scutellaria baicalensis might inhibit Kp biofilm formation through down-regulating integron geneⅠ1 expression.
Biofilms ; drug effects ; Integrons ; drug effects ; Klebsiella pneumoniae ; drug effects ; physiology ; Microbial Sensitivity Tests ; Scutellaria baicalensis

BACKGROUND/AIMS: To enable appropriate antimicrobial treatment for community-onset infections in emergency departments (EDs), data are needed on the resistance profiles of Escherichia coli and Klebsiella pneumoniae, which are the main pathogens of community-onset bacteremia. METHODS: Records were reviewed of 734 patients with E. coli and K. pneumoniae bacteremia who visited the Daegu Fatima Hospital ED, Daegu, Korea between 2003 and 2009. We investigated the demographic data, clinical findings, and antimicrobial susceptibility patterns of the organisms. RESULTS: Of 1,208 cases of community-onset bacteremia, 62.8% were caused by E. coli or K. pneumoniae in an ED of a secondary care hospital. Five hundred and forty-eight cases of E. coli (75%) and 183 cases of K. pneumoniae (25%) were analyzed. Urinary tract infection (43.1%) was most common, followed by intra-abdominal infection (39%) and pneumonia (7.2%). Trimethoprim/sulfamethoxazole, fluoroquinolone, third-generation cephalosporin (3GC) and amikacin resistance rates among E. coli and K. pneumoniae were 22.8%, 19.6%, 6.2%, and 1.3%, respectively. In 2009, the rate of 3GC resistance (10.6%) was significantly higher, compared to the annual averages of 2003 to 2008 (6.1%; p = 0.03). Previous exposure to antibiotics was an independent risk factor for 3GC resistance in multivariate logistic regression analysis. CONCLUSIONS: The rate of 3GC resistance increased in community-onset infections, and previous exposure to antibiotics was an independent risk factor. Despite the increased 3GC resistance in community-onset infections, an amikacin combination therapy could provide an option for treatment of bacteremic patients with previous antibiotic exposure in an ED.
Aged ; Aged, 80 and over ; Bacteremia/epidemiology/*microbiology ; *Cephalosporin Resistance ; Community-Acquired Infections/microbiology ; Emergency Service, Hospital/statistics & numerical data ; Escherichia coli/*physiology ; Female ; Humans ; Klebsiella pneumoniae/*physiology ; Male ; Middle Aged ; Republic of Korea/epidemiology ; Retrospective Studies ; Secondary Care Centers/statistics & numerical data

No abstract available.
Bacteremia/*microbiology ; *Cephalosporin Resistance ; Escherichia coli/*physiology ; Female ; Humans ; Klebsiella pneumoniae/*physiology ; Male

No abstract available.
Aged ; Drug Resistance, Bacterial ; Humans ; Imipenem/pharmacology/therapeutic use ; Klebsiella Infections/diagnosis/drug therapy/*microbiology ; Klebsiella pneumoniae/drug effects/isolation & purification/*physiology ; Male ; Microbial Sensitivity Tests ; Phenotype ; Sepsis/diagnosis/drug therapy/*microbiology ; Thienamycins/pharmacology/therapeutic use

We investigated the occurrence and genetic basis of AmpC beta-lactamase (AmpC)-mediated antibiotic resistance, by examining Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis isolates at a university hospital, from 2007 to 2010. The ampC genes were detected by multiplex AmpC PCR, and AmpC-positive strains were subjected to DNA sequencing. Extended-spectrum beta-lactamase (ESBL) production was assessed using the ESBL disk test based on the utilization of boronic acid. Carbapenem-resistant isolates were further investigated by the modified Hodge test, a carbapenemase inhibition test and SDS-PAGE experiments. AmpC expression was detected in 1.6% of E. coli (39 DHA-1, 45 CMY-2, and 1 CMY-1) isolates, 7.2% of K. pneumoniae (39 DHA-1, 45 CMY-2, and 1 CMY-1) isolates, and 2.5% of P. mirabilis (8 CMY-2 and 1 CMY-1) isolates. Of the 198 acquired AmpC producers, 58 isolates (29.3%) also produced an ESBL enzyme. Among the acquired AmpC-producing K. pneumoniae isolates, the minimum inhibitory concentration (MIC) MIC50/MIC90 values for cefoxitin, cefotaxime, cefepime, imipenem, and meropenem were >32/>32, 16/>32, 1/16, 0.25/0.5, and <0.125/0.125 microg/mL, respectively. The MIC values for carbapenem were > or =2 microg/mL for 2 K. pneumoniae isolates, both of which carried the blaDHA-1 gene with a loss of OmpK36 expression, but were negative for carbapenemase production. The acquisition of AmpC-mediated resistance in K. pneumoniae isolates increased, as did the proportion of AmpC and ESBL co-producers among the hospital isolates. The accurate identification of isolates producing AmpCs and ESBLs may aid in infection control and will assist physicians in selecting an appropriate antibiotic regimen.
Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/*genetics ; DNA, Bacterial/genetics ; Enterobacteriaceae Infections/*epidemiology/*microbiology ; Escherichia coli/drug effects/enzymology/isolation & purification ; Hospitals, University/statistics & numerical data ; Humans ; Klebsiella pneumoniae/drug effects/enzymology/isolation & purification/*physiology ; Microbial Sensitivity Tests ; Multiplex Polymerase Chain Reaction ; Proteus mirabilis/drug effects/enzymology/isolation & purification ; Republic of Korea/epidemiology ; beta-Lactamases/*genetics

OBJECTIVETo study the mechanism of protective effect of Jinqiaomai (JQM) on lung injury induced by Klebsiella pneumonia in rats.

METHODThe model of rats with Klebsiella pneumonia was established. The male SD rats were randomly divided into control group, model group, JQM (6, 3, 1.5 g x kg(-1)) three groups, levofloxacin (25 mg x kg(-1)) group, JQM (3 g x kg(-1)) + levofloxacin (25 mg x kg(-1)) group. The contents of IL-1beta, ICAM-1 and INF-gamma in the lung tissue homogenate were measured by radio-immunoassay and Elisa. TLR2/4 mRNA and MyD88 mRNA expression were detected by RT-PCR. IkappaB-alpha expression was detected by Western Blot.

RESULTThe rats of model group had obvious lung injury, but those of JQM, JQM + levofloxacin and levofloxacin groups had less injury. The contents of IL-1beta, ICAM-1 and INF-gamma in lung tissue homogenate and the expressions of TLR2/4 mRNA, MyD88 mRNA and IkappaB-alpha in lung of model group were significantly higher than those in the control group (P < 0.05 or P < 0.01), while IL-1beta, ICAM-1 and INF-gamma of JQM groups were significantly lower than those of model group (P < 0.05 or P < 0.01). The expressions of TLR2/4 mRNA, MyD88 mRNA and IkappaB-alpha of JQM (6.3 g x kg(-1)) groups were significantly lower than those of model group(P <0. 05 or P <0. 01).

CONCLUSIONThe lung injury induced by Klebsiella pneumonia is related to TLR2/4, MyD88 mRNA and IkappaB-alpha. To decrease the excessive expression of TLR2/4, MyD88 mRNA and IkappaB-alpha might be the main mechanism of protective effect of Jinqiaomai on lung injury.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Humans ; I-kappa B Proteins ; genetics ; metabolism ; Klebsiella Infections ; drug therapy ; genetics ; metabolism ; microbiology ; Klebsiella pneumoniae ; drug effects ; physiology ; Lung ; drug effects ; metabolism ; microbiology ; Male ; Myeloid Differentiation Factor 88 ; genetics ; metabolism ; NF-KappaB Inhibitor alpha ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism