This study aimed to provide evidence of persistent uropathogenic Escherichia coli (UPEC) in female patients with recurrent urinary tract infection (UTI) after antibiotic therapy. We collected biopsies of the bladder, and clean-catch urine samples from 32 women who had episodes of recurrent UTI and were given antibiotic therapy. Urine samples and biopsies were analyzed by conventional bacteriological techniques. Phylogenetic group and 16 virulence factors (VFs) of UPEC were determined using polymerase chain reaction (PCR). The infection capability of UPEC was confirmed in a mouse model. Immunofluorescence and electron microscopy were used to detect intracellular bacterial communities (IBCs) in the mouse model. The results showed that all urine specimens were detected sterile. E. coli was found in 6 of 32 biopsies (18.75%), and was identified to be UPEC by PCR. Different VFs associated with the formation of IBCs were identified in all six UPEC isolates. Each UPEC isolate was capable of forming IBCs within the bladder epithelial cells of mice. In conclusion, UPEC with distinctive pathological traits and the capability of IBC formation was first found in the bladders of women after antibiotic therapy, suggesting that the IBC pathogenic pathway may occur in humans and it plays an important role in UTI recurrence.
Adult ; Animals ; Anti-Bacterial Agents ; administration & dosage ; Biopsy ; Escherichia coli Infections ; drug therapy ; microbiology ; Female ; Humans ; Mice ; Middle Aged ; Phylogeny ; Urinary Bladder ; drug effects ; microbiology ; pathology ; Uropathogenic Escherichia coli ; drug effects ; genetics ; pathogenicity

Urinary tract infection (UTI) is one of the most common complications after radical cystectomy and orthotopic neobladder reconstruction. This study investigated the incidence and implicated pathogen of febrile UTI after ileal neobladder reconstruction and identify clinical and urodynamic parameters associated with febrile UTI. From January 2001 to May 2015, 236 patients who underwent radical cystectomy and ileal neobladder were included in this study. Fifty-five episodes of febrile UTI were identified in 46 patients (19.4%). The probability of febrile UTI was 17.6% and 19.8% at 6 months and 24 months after surgery, respectively. While, Escherichia coli was the most common implicated pathogen (22/55, 40.0%), Enterococcus spp. were the most common pathogen during the first month after surgery (18/33, 54.5%). In multivariate logistic regression analysis, ureteral stricture was an independent risk factor associated with febrile UTI (OR 5.93, P = 0.023). However, ureteral stricture accounted for only 6 episodes (10.9%, 6/55) of febrile UTI. Most episodes of febrile UTI occurred within 6 months after surgery. Thus, to identify risk factors associated with febrile UTI in the initial postoperative period, we assessed videourodynamics within 6 months after surgery in 38 patients. On videourodyamic examination, vesicoureteral reflux (VUR) was identified in 16 patients (42.1%). The rate of VUR presence in patients who had febrile UTI was not significantly different from those in patients without febrile UTI (50% vs. 39.3%, P = 0.556). Patients with febrile UTI had significantly larger residual urine volume (212.0 ± 193.7 vs. 90.5 ± 148.2, P = 0.048) than those without. E. coli and Enterococcus spp. are common pathogens and ureteral stricture and residual urine are risk factors for UTI after ileal neobladder reconstruction.
Aged ; Anti-Bacterial Agents/therapeutic use ; Cystectomy/adverse effects ; Enterococcus/isolation & purification ; Escherichia coli/isolation & purification ; Female ; Humans ; Ileum/*surgery ; Incidence ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Postoperative Complications ; Reconstructive Surgical Procedures ; Retrospective Studies ; Risk Factors ; Urinary Bladder Neoplasms/*surgery ; Urinary Tract Infections/drug therapy/*epidemiology/etiology/microbiology ; Urodynamics

OBJECTIVETo evaluate the value of urine sediment analyzer in the screening of clinically suspected urinary tract infection (UTI) in cancer patients.

METHODSThe results of bacterial count of 1 053 midstream urine samples by UF-1000i urine sediment analyzer (UF-1000i urine sediment analyzer, UF-1000i) were compared with the results of bacterial culture. Moreover, the results of distinguishing bacterial species by the bacterial scattergram were compared with the results of bacteria culture. At the same time, the sensitivity, specificity, positive predictive value and negative predictive value of UF-1000i analyzer for UTI screening were evaluated.

RESULTSOf all the 1 053 samples, the top three bacteria were E. coli, Enterococci and P. aeruginosa. The top three malignant tumors of UTI were bladder, lung cancer and cervical cancers. The positive rate of UF-1000i analyzer was 20% (211/1 053), and that of bacteria culture was 17.9% (188/1 053). There was statistically no significant difference in the positive rates between the two methods (χ(2)=1.636, P>0.05), and the two methods had a considerable consistency (Kappa=0.756). Compared with the clinical diagnosis, UTI screening by UF-1000i analyzer showed a sensitivity of 79.6% (160/201), specificity of 95.5% (814/852), positive predictive value of 80.8% (160/198) and negative predictive value of 95.2%(814/855). The distribution of cocci and bacilli acquired by the bacterial scattergram was basically in accordance with the results of bacterial culture.

CONCLUSIONSBacterial count by UF-1000i analyzer plays an important role in early screening of UTI, and the bacterial scattergram may help to distinguish bacterial species, providing reference for the use of antibiotics in early medication.

Bacterial Load ; Enterococcus ; isolation & purification ; Escherichia coli ; isolation & purification ; Female ; Flow Cytometry ; Humans ; Leukocyte Count ; Lung Neoplasms ; urine ; Predictive Value of Tests ; Pseudomonas aeruginosa ; isolation & purification ; Sensitivity and Specificity ; Urinary Bladder Neoplasms ; urine ; Urinary Tract Infections ; diagnosis ; microbiology ; urine ; Uterine Cervical Neoplasms ; urine

No abstract available.
Aged ; Anti-Bacterial Agents/therapeutic use ; Bacteria/metabolism ; Catheter-Related Infections/diagnosis/drug therapy/*microbiology ; Color ; Equipment Design ; Escherichia coli Infections/diagnosis/drug therapy/*microbiology ; Female ; Humans ; Intestines/*microbiology ; Pigments, Biological/metabolism ; Treatment Outcome ; Tryptophan/metabolism ; Urinary Bladder Neoplasms/surgery ; Urinary Catheterization/adverse effects/*instrumentation ; *Urinary Catheters ; *Urinary Diversion ; Urinary Tract Infections/diagnosis/drug therapy/*microbiology ; Urine/chemistry/microbiology

OBJECTIVETo study the role of outer membrane protein T (OmpT) in the pathogenesis of uropathogenic Escherichia.coli.

METHODSIn cultured human bladder epithelial cell line 5637, we examined the adhesion ability of wild-type (CFT073), ompT gene knockout (COTD), and revertant (pST) strains of E.coli to the cells and the extracellular matrix (ECM). The expressions of the adhesion gene iha and virulence gene iroN were detected by real-time PCR. Murine models of urinary tract infection with the 3 strains were established to evaluate the bacterial burden of the bladder and kidney tissue and bacterial counts in blood. We also detected the expressions of interleukin-6 (IL-6) and IL-8 in the bladder and kidney tissues of the mice.

RESULTThe COTD strain showed a significantly lower cell adhesion rate than CFT073 strain [(4.62∓0.39)% vs (8.81∓1.13)%, P<0.05] with also a lower ECM-adhesion rate [(4.95∓0.59)% vs (8.85∓0.79)%, P<0.05]. The mRNA expressions of iha and iroN in CFT073 strain were 2.1 and 3.8 times that of COTD strain. In the mouse model, the mean bacterial load of CFT073 strain in the bladder tissue was 6.36∓0.06, significantly greater than that of COTD (6.01∓0.07) and revertant (6.29∓0.06) strains (P<0.05); the bacterial load of CFT073 strain in the kidney tissue was also significantly higher than that of COTD strain (6.25∓0.05 vs 5.87∓0.06, P<0.05). In mice infected with the wild-type, knockout, and revertant strains, the detection rates of IL-6, which were identical to those of IL-8, in the inflammatory bladder and kidney tissues were 60%, 12.5%, and 50%, respectively.

CONCLUSIONSOmpT may regulate the expression of the adhesion gene iha and the transferrin gene iroN to affect the adhesion of uropathogenic E.coli to host cells.

Animals ; Bacterial Adhesion ; Bacterial Load ; Bacterial Outer Membrane Proteins ; metabolism ; Cell Line, Tumor ; Escherichia coli Infections ; pathology ; Escherichia coli Proteins ; metabolism ; Gene Knockout Techniques ; Humans ; Inflammation ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Kidney ; microbiology ; Mice ; Peptide Hydrolases ; metabolism ; Receptors, Cell Surface ; metabolism ; Urinary Bladder ; microbiology ; Urinary Tract Infections ; microbiology ; pathology ; Uropathogenic Escherichia coli ; pathogenicity

OBJECTIVETo detect the effects of Polyporus polysaccharide (PPS), Bacillus Calmette-Guerin (BCG), and their combination on the nuclear factor kappa B (NF-κB) signaling pathway associated-gene expression and investigate the molecular mechanisms of the toxic-reducing effect of PPS in coordination with BCG against bladder cancer.

METHODSAfter T739 cells were treated with PPS, BCG and their combination, the changes in mRNA and protein expression of inhibitor of kappa B kinase beta (IKKβ), NF-κB subunit p65 (NF-κB p65), intracellular adhesion molecule 1 (ICAM1) and chemokine (C-c motif) ligand 2 (CCL2) in bladder cancer cell line T739 were determined by relative quantitative real-time PCR, Western blot, and flow cytometry (FCM). NF-κB p65 DNA-binding activity in T739 cell was detected by biotinylated probe-ELISA, and NF-κB p65 nuclear expression in T739 cell was observed by immunohistochemistry.

RESULTSCompared with the T739 control group, the mRNA expression of IKBKB (IKKβ), Rel A (NF-κB p65), ICAM1 and CCL2 in T739 cells treated with BCG were increased obviously (Ratio>2.0), as well as the expression of IKKβ, CCL2 and ICAM1 proteins. Meanwhile, NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression in T739 cells treated with BCG were up-regulated significantly (P<0.05). Compared with the control, the increased expression in T739 cells were simultaneously down-regulated after PPS treatment, except for ICAM1 protein expression. With cells treated with a combination of BCG and PPS, the expression of genes associated with the NF-κB signaling pathway, such as IKBKB, ICAM1 and CCL2, were all down-regulated compared to the BCG group, as well as Rel A mRNA expression, NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression.

CONCLUSIONSPPS could inhibit the over-activation of the NF-κB signaling pathway induced by BCG in bladder cancer cells and accordingly attenuate the adverse reactions to BCG therapy.

Cell Line, Tumor ; Cell Nucleus ; drug effects ; metabolism ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Mycobacterium bovis ; drug effects ; NF-kappa B ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Polyporus ; chemistry ; Polysaccharides ; pharmacology ; Signal Transduction ; drug effects ; Urinary Bladder Neoplasms ; genetics ; microbiology ; pathology

To study the relationship between bacterial infection and the etiology of cystitis glandularis, 36 female Wistar rats were divided into 3 groups. No intervention was given to the rats in the blank group. NS was infused into the bladder of the rats of the control group, and solution containing E. coli was injected into the bladder of experimental group. Three months later, tissue samples of bladder were collected and observed visually and under light microscope. The results showed that tissues of the blank group were normal; one sample in the control group showed Brunn's nests and cystitis cystica, and 10 in the experimental group had the change of cystitis glandularis. Compared to the blank and control group, samples in the experimental group showed significant change (P<0.05). There were no significant difference between blank group and control group (P>0.05). It is concluded that bladder instillation of E. coli can induce cystitis glandularis, which confirms that infection is the cause of cystitis glandularis.
Cystitis/etiology ; Cystitis/*microbiology ; Cystitis/*pathology ; Escherichia coli Infections/*pathology ; Rats, Wistar ; Urinary Bladder/microbiology ; Urinary Bladder/*pathology ; Urinary Tract Infections/complications ; Urinary Tract Infections/*microbiology ; Urinary Tract Infections/pathology

OBJECTIVETo investigate whether different types of injury on bladder wall can influence bacillus Calmette-Guerin (BCG) attachment.

METHODSThe bladder mucosa of 24 rabbits were treated by electrocautery,cryocautery and incision on left lateral wall, right lateral wall and posterior wall, respectively. Then radiolabeled BCG ((3)H-BCG) was instilled into bladder. Two hours latter, the injured bladder wall with different methods and non-injured wall (anterior wall of bladder) were surgically removed and digested. The quantity of BCG of each specimen was determined by liquid scintillation counter.

RESULTThe quantity of BCG attachment to bladder wall with different injuries was significantly higher than that of non-injured wall (P<0.001), meanwhile there was no statistically difference among the BCG levels of different injury types (P>0.05).

CONCLUSIONBCG attachment is not influenced by different types of injury on the bladder wall.

Animals ; Bacterial Adhesion ; Female ; Male ; Mycobacterium bovis ; physiology ; Rabbits ; Urinary Bladder ; injuries ; microbiology

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 ; Cystitis ; etiology ; microbiology ; pathology ; Escherichia coli Infections ; Female ; Humans ; Male ; Metaplasia ; Middle Aged ; Urinary Bladder ; microbiology ; pathology ; Urinary Tract Infections ; complications ; microbiology ; pathology

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.
Cystitis/etiology ; Cystitis/microbiology ; Cystitis/*pathology ; *Escherichia coli Infections ; Metaplasia ; Urinary Bladder/microbiology ; Urinary Bladder/*pathology ; Urinary Tract Infections/complications ; Urinary Tract Infections/*microbiology ; Urinary Tract Infections/pathology